PMID- 16210303 TI - Working time directive shift patterns may improve care. PMID- 16210304 TI - Which career first? Economic perspective on later pregnancy is positive. PMID- 16210305 TI - Which career first? Financial and social reasons should have been taken into account. PMID- 16210306 TI - Which career first? Women don't want it all, but they may want children. PMID- 16210307 TI - Which career first? Skewed argument should be put to bed. PMID- 16210308 TI - Which career first? Women remain caught in dilemma. PMID- 16210309 TI - Smoking behaviour in European patients with established coronary heart disease. AB - AIMS: Treatment guidelines for patients with established coronary disease emphasize the importance of smoking cessation. We aimed to study smoking behaviour in European patients, as well as trends in this behaviour over time. We further aimed to evaluate the relation between selected patient characteristics and smoking cessation. METHODS AND RESULTS: We studied patients who were enrolled in the second European Action on Secondary Prevention through Intervention to Reduce Events (EUROASPIRE) survey, which was undertaken in 15 European countries during 1999-2000. Consecutive patients, < or =70 years were identified after coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, admission for myocardial infarction, or admission for myocardial ischaemia. Patients were then interviewed, at a median of 1.5 years after the index event. During the interview, data were collected on a broad range of clinical characteristics, including smoking status, which was validated by breath carbon monoxide levels. The prevalence of smoking was compared with data from the first EUROASPIRE survey, which had a similar design, and was conducted during 1995-96. In EUROASPIRE II, 5551 patients attended the interview and 1172 (21%) were (persistent) smokers. No decrease in smoking prevalence was observed similar to the first survey (19%). The proportion of smokers was 39% in patients aged <50, 26% in patients aged 50-60, and 14% in patients aged > or =60. Men and women had similar prevalence. A verbal advice to give up smoking by a medical professional was given to 99% of the 2244 pre-event smokers, and 48% actually stopped. This proportion was relatively high in elderly patients > or =60 years (53%), patients with a university level of education (56%), and patients with a myocardial infarction as the index event (52%). The proportion of stopped smokers was low in patients <50 years (41%) and in patients with myocardial ischaemia as the index event (38%). CONCLUSION: The prevalence of smoking in European patients with established coronary disease is too high: one out of each five patients smokes, despite a personal advice to stop. Thus, still there is a need for the development of effective smoking cessation programmes. PMID- 16210310 TI - Comparative evaluation of the cerebral state index and the bispectral index during target-controlled infusion of propofol. AB - BACKGROUND: Cerebral state index (CSI) has recently been introduced as an intra operative monitor of anaesthetic depth. We compared the performance of the CSI to the bispectral index (BIS) in measuring depth of anaesthesia during target controlled infusion (TCI) of propofol. METHODS: Twenty Chinese patients undergoing general anaesthesia were recruited. CSI and BIS, and predicted effect site concentration of propofol were recorded. The level of sedation was tested by Modified Observer's Assessment of Alertness/Sedation Scale (MOAAS) every 20 s during stepwise increase (TCI, 0.5 microg ml(-1)) of propofol. The loss of verbal contact (LVC) and loss of response (LOR) were defined by MOAAS values of 2-3 and less than 2, respectively. Baseline variability and the prediction probability (P(K)) were calculated for the BIS and CSI. The values of BIS(05) and CSI(05), BIS(50) and CSI(50), BIS(95) and CSI(95) were calculated at each end-point (LVC and LOR). RESULTS: Baseline variability of CSI was more than that of BIS. Both CSI and BIS showed a high prediction probability for the steps awake vs LVC, awake vs LOR, and LVC vs LOR, and good correlations with MOAAS values. CONCLUSION: Despite larger baseline variation, CSI performed as well as BIS in terms of P(K) values and correlations with step changes in sedation. PMID- 16210311 TI - A randomized non-crossover study comparing the ProSeal and Classic laryngeal mask airway in anaesthetized children. AB - BACKGROUND: We tested the hypothesis that ease of insertion, oropharyngeal leak pressure, fibreoptic position, gastric insufflation, and the frequency of mucosal trauma differ between the ProSeal laryngeal mask airway (PLMA) and the classic laryngeal mask airway (cLMA) in anaesthetized children. For the PLMA, we also assessed the ease of gastric tube placement via the PLMA drain tube and measure residual gastric volume. METHODS: 240 consecutive ASA I-III children aged 1-16 yr were randomized for airway management with the ProSeal or cLMA. RESULTS: The time taken to provide an effective airway, the number of insertion attempts, fibreoptic position of the airway tube and frequency of mucosal trauma were similar, but oropharyngeal leak pressure was higher (33 vs 26 cm H(2)O, P<0.0001) and gastric insufflation less common (0 vs 6%, P<0.01) for the PLMA. Gastric tube insertion was successful at the first attempt in 106 of 120, and at the second attempt in 14 of 120. The mean (sd; range) value for residual gastric volume was 2.2 (5.9; 0-30) ml. There were no differences in performance among sizes for the PLMA and the cLMA. CONCLUSIONS: We conclude that ease of insertion, fibreoptic position, and frequency of mucosal trauma are similar for the PLMA and cLMA in children, but oropharyngeal leak pressure is higher and gastric insufflation less common for the PLMA. Gastric tube insertion has a high success rate, provided the PLMA is correctly positioned. PMID- 16210312 TI - Accommodation and repair of a UV photoproduct in DNA at different rotational settings on the nucleosome surface. AB - Cyclobutane-thymine dimers (CTDs), the most common DNA lesion induced by UV radiation, cause 30 degrees bending and 9 degrees unwinding of the DNA helix. We prepared site-specific CTDs within a short sequence bracketed by strong nucleosome-positioning sequences. The rotational setting of CTDs over one turn of the helix near the dyad center on the histone surface was analyzed by hydroxyl radical footprinting. Surprisingly, the position of CTDs over one turn of the helix does not affect the rotational setting of DNA on the nucleosome surface. Gel-shift analysis indicates that one CTD destabilizes histone-DNA interactions by 0.6 or 1.1 kJ/mol when facing away or toward the histone surface, respectively. Thus, 0.5 kJ/mol energy penalty for a buried CTD is not enough to change the rotational setting of sequences with strong rotational preference. The effect of rotational setting on CTD removal by nucleotide excision repair (NER) was examined using Xenopus oocyte nuclear extracts. The NER rates are only 2-3 times lower in nucleosomes and change by only 1.5-fold when CTDs face away or toward the histone surface. Therefore, in Xenopus nuclear extracts, the rotational orientation of CTDs on nucleosomes has surprisingly little effect on rates of repair. These results indicate that nucleosome dynamics and/or chromatin remodeling may facilitate NER in gaining access to DNA damage in nucleosomes. PMID- 16210313 TI - The sequence at specific positions in the early transcribed region sets the rate of transcript synthesis by RNA polymerase II in vitro. AB - To further understand the mechanism of promoter escape by RNA polymerase II, we have systematically investigated the effect of core promoter sequence on the rate of transcript synthesis in vitro. Chimeric and mutant promoters were made by swapping sequences between the human interleukin-2 promoter and the adenovirus major late promoter, which exhibit different rates of transcript synthesis. Kinetic studies at these promoters revealed that sequences downstream of the start sites set the rate of transcript synthesis. Specifically, the sequences at +2 and +7/+8 are critical for determining the rate; when either +2 is a C (nontemplate strand) or +7/+8 is a TT (nontemplate strand), transcript synthesis is slow. At +7/+8, the thermodynamic stability of the RNA:DNA hybrid controls the overall rate of transcript synthesis. Our data support a model in which the rate limiting step during transcript synthesis by RNA polymerase II in vitro occurs at the point in the reaction at which early ternary complexes transform into elongation complexes. PMID- 16210314 TI - The C-terminal RpoN domain of sigma54 forms an unpredicted helix-turn-helix motif similar to domains of sigma70. AB - The "sigma" subunit of prokaryotic RNA polymerase allows gene-specific transcription initiation. Two sigma families have been identified, sigma70 and sigma54, which use distinct mechanisms to initiate transcription and share no detectable sequence homology. Although the sigma70-type factors have been well characterized structurally by x-ray crystallography, no high resolution structural information is available for the sigma54-type factors. Here we present the NMR-derived structure of the C-terminal domain of sigma54 from Aquifex aeolicus. This domain (Thr-323 to Gly-389), which contains the highly conserved RpoN box sequence, consists of a poorly structured N-terminal tail followed by a three-helix bundle, which is surprisingly similar to domains of the sigma70-type proteins. Residues of the RpoN box, which have previously been shown to be critical for DNA binding, form the second helix of an unpredicted helix-turn helix motif. The homology of this structure with other DNA-binding proteins, combined with previous biochemical data, suggests how the C-terminal domain of sigma54 binds to DNA. PMID- 16210315 TI - Discovery and characterization of the cryptic psi subunit of the pseudomonad DNA replicase. AB - We previously reconstituted a minimal DNA replicase from Pseudomonas aeruginosa consisting of alpha and epsilon (polymerase and editing nuclease), beta (processivity factor), and the essential tau, delta, and delta' components of the clamp loader complex (Jarvis, T., Beaudry, A., Bullard, J., Janjic, N., and McHenry, C. (2005) J. Biol. Chem. 280, 7890-7900). In Escherichia coli DNA polymerase III holoenzyme, chi and Psi are tightly associated clamp loader accessory subunits. The addition of E. coli chiPsi to the minimal P. aeruginosa replicase stimulated its activity, suggesting the existence of chi and Psi counterparts in P. aeruginosa. The P. aeruginosa chi subunit was recognizable from sequence similarity, but Psi was not. Here we report purification of an endogenous replication complex from P. aeruginosa. Identification of the components led to the discovery of the cryptic Psi subunit, encoded by holD. P. aeruginosa chi and Psi were co-expressed and purified as a 1:1 complex. P. aeruginosa chiPsi increased the specific activity of tau(3)deltadelta' 25-fold and enabled the holoenzyme to function under physiological salt conditions. A synergistic effect between chiPsi and single-stranded DNA binding protein was observed. Sequence similarity to P. aeruginosa Psi allowed us to identify Psi subunits from several other Pseudomonads and to predict probable translational start sites for this protein family. This represents the first identification of a highly divergent branch of the Psi family and confirms the existence of Psi in several organisms in which Psi was not identifiable based on sequence similarity alone. PMID- 16210316 TI - Activation of the Src family kinase Hck without SH3-linker release. AB - Src family protein-tyrosine kinases are regulated by intramolecular binding of the SH2 domain to the C-terminal tail and association of the SH3 domain with the SH2 kinase-linker. The presence of two regulatory interactions raises the question of whether disruption of both is required for kinase activation. To address this question, we engineered a high affinity linker (HAL) mutant of the Src family member Hck in which an optimal SH3 ligand was substituted for the natural linker. Surface plasmon resonance analysis demonstrated tight intramolecular binding of the modified HAL sequence to SH3. Hck-HAL was then combined with a tail tyrosine mutation (Y501F) and expressed in Rat-2 fibroblasts. Surprisingly, Hck-HAL-Y501F showed strong transforming and kinase activities, demonstrating that intramolecular SH3-linker release is not required for SH2-based kinase activation. In Saccharomyces cerevisiae, which lacks the negative regulatory tail kinase Csk, wild-type Hck was more strongly activated in the presence of an SH3-binding protein (human immunodeficiency virus-1 Nef), indicating persistence of native SH3-linker interaction in an active Hck conformation. Taken together, these data support the existence of multiple active conformations of Src family kinases that may generate unique downstream signals. PMID- 16210318 TI - p-Hydroxybenzoic acid synthesis in Mycobacterium tuberculosis. AB - Glycosylated p-hydroxybenzoic acid methyl esters and structurally related phenolphthiocerol glycolipids are important virulence factors of Mycobacterium tuberculosis. Although both types of molecules are thought to be derived from p hydroxybenzoic acid, the origin of this putative biosynthetic precursor in mycobacteria remained to be established. We describe the characterization of a transposon mutant of M. tuberculosis deficient in the production of all forms of p-hydroxybenzoic acid derivatives. The transposon was found to be inserted in Rv2949c, a gene located in the vicinity of the polyketide synthase gene pks15/1, involved in the elongation of p-hydroxybenzoate to phenolphthiocerol in phenolic glycolipid-producing strains. A recombinant form of the Rv2949c enzyme was produced in the fast-growing non-pathogenic Mycobacterium smegmatis and purified to near homogeneity. The recombinant enzyme catalyzed the removal of the pyruvyl moiety of chorismate to form p-hydroxybenzoate with an apparent K(m) value for chorismate of 19.7 microm and a k(cat) value of 0.102 s(-1). Strong inhibition of the reaction by p-hydroxybenzoate but not by pyruvate was observed. These results establish Rv2949c as a chorismate pyruvate-lyase responsible for the direct conversion of chorismate to p-hydroxybenzoate and identify Rv2949c as the sole enzymatic source of p-hydroxybenzoic acid in M. tuberculosis. PMID- 16210317 TI - Cell-specific regulation of androgen receptor phosphorylation in vivo. AB - The biological ramifications of phosphorylation of the androgen receptor (AR) are largely unknown. To examine the phosphorylation of AR at serine 213, a putative substrate for Akt, a phosphorylation site-specific antibody was generated. The use of this antibody indicated that AR Ser-213 is phosphorylated in vivo and that phosphorylation is tightly regulated in a cell type-specific manner. Furthermore, Ser-213 phosphorylation took place with rapid kinetics and was inhibited by the phosphatidylinositol 3-kinase inhibitor LY294002. Phosphorylation occurred in response to R1881 and dihydrotestosterone but weakly if at all in response to testosterone. It did not occur in response to AR antagonists or growth factor stimulation in the absence of an AR agonist. Transcription assays using an AR responsive reporter gene construct showed that activated phosphatidylinositol 3 kinase inhibited transcription mediated by wild type AR but not that of a mutant AR variant (S213A), which could not be phosphorylated at Ser-213. By immunohistochemistry, the AR Ser(P)-213 antigen was detected in prostate epithelial but not stromal cells despite the fact that an antibody recognizing both phosphorylated and non-phosphorylated forms of AR demonstrates that AR is present in both cell types as expected. In fetal tissue the AR-Ser(P)-213 antigen was present in epithelial cells of the urogenital sinus when endogenous androgen levels were high and activated Akt was prevalent, but absent at a later stage of development when endogenous androgen levels were low and Akt activation was minimal. Immunoreactivity was evident in differentiated cells lining the lumen of the urogenital sinus but not in rapidly dividing, Ki67 positive cells within the developing prostate or stromal tissue, suggesting that site-specific phosphorylation of AR Ser-213 by cellular kinases occurs in a non-proliferating cellular milieu. PMID- 16210319 TI - Distinct roles for SCL in erythroid specification and maturation in zebrafish. AB - The stem cell leukemia (SCL) transcription factor is essential for vertebrate hematopoiesis. Using the powerful zebrafish model for embryonic analysis, we compared the effects of either reducing or ablating Scl using morpholino-modified antisense RNAs. Ablation of Scl resulted in the loss of primitive and definitive hematopoiesis, consistent with its essential role in these processes. Interestingly, in embryos with severely reduced Scl levels, erythroid progenitors expressing gata1 and embryonic globin developed. Erythroid maturation was deficient in these Scl hypomorphs, supporting that Scl was required both for the erythroid specification and for the maturation steps, with maturation requiring higher Scl levels than specification. Although all hematopoietic functions were rescued by wild-type Scl mRNA, an Scl DNA binding mutant rescued primitive and definitive hematopoiesis but did not rescue primitive erythroid maturation. Together, we showed that there is a distinct Scl hypomorphic phenotype and demonstrated that distinct functions are required for the roles of Scl in the specification and differentiation of primitive and definitive hematopoietic lineages. Our results revealed that Scl participates in multiple processes requiring different levels and functions. Further, we identified an Scl hypomorphic phenotype distinct from the null state. PMID- 16210320 TI - All four putative selectivity filter glycine residues in KtrB are essential for high affinity and selective K+ uptake by the KtrAB system from Vibrio alginolyticus. AB - The subunit KtrB of bacterial Na+-dependent K+-translocating KtrAB systems belongs to a superfamily of K+ transporters. These proteins contain four repeated domains, each composed of two transmembrane helices connected by a putative pore loop (p-loop). The four p-loops harbor a conserved glycine residue at a position equivalent to a glycine selectivity filter residue in K+ channels. We investigated whether these glycines also form a selectivity filter in KtrB. The single residues Gly70, Gly185, Gly290, and Gly402 from p-loops P(A) to P(D) of Vibrio alginolyticus KtrB were replaced with alanine, serine, or aspartate. The three alanine variants KtrB(A70), KtrB(A185), and KtrB(A290) maintained a substantial activity in KtrAB-mediated K+ uptake in Escherichia coli. This activity was associated with a decrease in the affinity for K+ by 2 orders of magnitude, with little effect on Vmax. Minor activities were also observed for three other variants: KtrB(A402), KtrB(S70), and KtrB(D185). With all of these variants, the property of Na+ dependence of K+ transport was preserved. Only the four serine variants mediated Na+ uptake, and these variants differed considerably in their K+/Na+ selectivity. Experiments on cloned ktrB in the pBAD18 vector showed that V. alginolyticus KtrB alone was still active in E. coli. It mediated Na+-independent, slow, high affinity, and mutation-specific K+ uptake as well as K+-independent Na+ uptake. These data demonstrate that KtrB contains a selectivity filter for K+ ions and that all four conserved p-loop glycine residues are part of this filter. They also indicate that the role of KtrA lies in conferring velocity and ion coupling to the Ktr complex. PMID- 16210321 TI - A transgenic mouse model of heart failure using inducible Galpha q. AB - Receptors coupled to Galpha q play a key role in the development of heart failure. Studies using genetically modified mice suggest that Galpha q mediates a hypertrophic response in cardiac myocytes. Galpha q signaling in these models is modified during early growth and development, whereas most heart failure in humans occurs after cardiac damage sustained during adulthood. To determine the phenotype of animals that express increased Galpha q signaling only as adults, we generated transgenic mice that express a silent Galpha q protein (Galpha qQ209L hbER) in cardiac myocytes that can be activated by tamoxifen. Following drug treatment to activate Galpha q Q209L-hbER, these mice rapidly develop a dilated cardiomyopathy and heart failure. This phenotype does not appear to involve myocyte hypertrophy but is associated with dephosphorylation of phospholamban (PLB), decreased sarcoplasmic reticulum Ca2+-ATPase activity, and a decrease in L type Ca2+ current density. Changes in Ca2+ handling and decreased cardiac contractility are apparent 1 week after Galpha qQ209L-hbER activation. In contrast, transgenic mice that express an inducible Galpha q mutant that cannot activate phospholipase Cbeta (PLCbeta) do not develop heart failure or changes in PLB phosphorylation, but do show decreased L-type Ca2+ current density. These results demonstrate that activation of Galpha q in cardiac myocytes of adult mice causes a dilated cardiomyopathy that requires the activation of PLCbeta. However, increased PLCbeta signaling is not required for all of the Galpha q-induced cardiac abnormalities. PMID- 16210322 TI - Arachidonic acid inhibits the insulin induction of glucose-6-phosphate dehydrogenase via p38 MAP kinase. AB - Polyunsaturated fatty acids are potent inhibitors of lipogenic gene expression in liver. The lipogenic enzyme glucose-6-phosphate dehydrogenase (G6PD) is unique in this gene family, in that fatty acids inhibit at a post-transcriptional step. In this study, we have provided evidence for a signaling pathway for the arachidonic acid inhibition of G6PD mRNA abundance. Arachidonic acid decreases the insulin induction of G6PD expression; by itself, arachidonic acid does not inhibit basal G6PD mRNA accumulation. The insulin stimulation of G6PD involves the phosphoinositide 3-kinase (PI 3-kinase) pathway (Wagle, A., Jivraj, S., Garlock, G. L., and Stapleton, S. R. (1998) J. Biol. Chem. 273, 14968-14974). Incubation of hepatocytes with arachidonic acid blocks the activation of PI 3-kinase by insulin as observed by a decrease in Ser(473) phosphorylation of Akt, the downstream effector of PI 3-kinase. The decrease in PI 3-kinase activity was associated with an increase in Ser(307) phosphorylation of IRS-1. Western analysis demonstrated increased phosphorylation of p38 mitogen-activated protein kinase (MAPK) in arachidonic acid-treated cells, whereas extracellular signal regulated kinase and c-Jun NH(2)-terminal kinase activity was not changed. Incubating the hepatocytes with the p38 MAPK inhibitor, SB203580, blocked the arachidonic acid inhibition of G6PD mRNA accumulation. Furthermore, SB203580 decreased the arachidonic acid-mediated Ser(307) phosphorylation of IRS-1 and rescued Akt activation that was otherwise decreased by arachidonic acid. Thus, arachidonic acid inhibits the insulin stimulation of G6PD mRNA accumulation by stimulating the p38 MAPK pathway, thereby inhibiting insulin signal transduction. PMID- 16210323 TI - Geldanamycin induces heat shock protein 70 and protects against MPTP-induced dopaminergic neurotoxicity in mice. AB - As key molecular chaperone proteins, heat shock proteins (HSPs) represent an important cellular protective mechanism against neuronal cell death in various models of neurological disorders. In this study, we investigated the effect as well as the molecular mechanism of geldanamycin (GA), an inhibitor of Hsp90, on 1 methyl-4-pheny-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurotoxicity, a mouse model of Parkinson disease. Neurochemical analysis showed that pretreatment with GA (via intracerebral ventricular injection 24 h prior to MPTP treatment) increased residual dopamine content and tyrosine hydroxylase immunoreactivity in the striatum 24 h after MPTP treatment. To dissect out the molecular mechanism underlying this neuroprotection, we showed that the GA mediated protection against MPTP was associated with a reduction of cytosolic Hsp90 and an increase in Hsp70, with no significant changes in Hsp40 and Hsp25 levels. Furthermore, in parallel with the induction of Hsp70, striatal nuclear HSF1 levels and HSF1 binding to heat shock element sites in the Hsp70 promoter were significantly enhanced by the GA pretreatment. Together these results suggested that the molecular cascade leading to the induction of Hsp70 is critical to the neuroprotection afforded by GA against MPTP-induced neurotoxicity in the brain and that pharmacological inhibition of Hsp90 may represent a potential therapeutic strategy for Parkinson disease. PMID- 16210324 TI - Identification and characterization of a novel human type II diacylglycerol kinase, DGK kappa. AB - Diacylglycerol kinase (DGK) plays an important role in signal transduction through modulating the balance between two signaling lipids, diacylglycerol and phosphatidic acid. Here we identified a tenth member of the DGK family designated DGK kappa. The kappa-isozyme (1271 amino acids, calculated molecular mass, 142 kDa) contains a pleckstrin homology domain, two cysteine-rich zinc finger-like structures, and a separated catalytic region as have been found commonly for the type II isozymes previously cloned (DGKdelta and DGKeta). The new DGK isozyme has additionally 33 tandem repeats of Glu-Pro-Ala-Pro at the N terminus. Reverse transcriptase-PCR showed that the DGK kappa mRNA is most abundant in the testis, and to a lesser extent in the placenta. DGK kappa, when expressed in HEK293 cells, was persistently localized at the plasma membrane even in the absence of cell stimuli. Deletion analysis revealed that the short C-terminal sequence (amino acid residues 1199-1268) is necessary and sufficient for the plasma membrane localization. Interestingly, DGK kappa, but not other type II DGKs, was specifically tyrosine-phosphorylated at Tyr78 through the Src family kinase pathway in H2O2-treated cells. Moreover, H2O2 selectively inhibited DGK kappa activity in a Src family kinase-independent manner, suggesting that the isozyme changes the balance of signaling lipids in the plasma membrane in response to oxidative stress. The expression patterns, subcellular distribution, and regulatory mechanisms of DGK kappa are distinct from those of DGKdelta and DGKeta despite high structural similarity, suggesting unique functions of the individual type II isozymes. PMID- 16210325 TI - Mechanisms of cell-surface rerouting of an endoplasmic reticulum-retained mutant of the vasopressin V1b/V3 receptor by a pharmacological chaperone. AB - Cell-surface expression and biological functions of several intracellular retained G protein-coupled receptors are restored by membrane-permeable ligands called pharmacological chaperones. We have previously demonstrated that a mutation of the hydrophobic motif 341FNX2LLX3L350 in the C terminus of the human pituitary vasopressin V3 receptor (MUT V3R) led to it being retained in the endoplasmic reticulum (ER). Here, we establish the precise role of this motif and investigate whether SSR149415, a non-peptide V3R antagonist, behaves as a pharmacological chaperone for the ER-retained MUT V3R. The absence of the mutated receptor in the plasma membrane is linked to its prolonged association with the molecular chaperone calnexin in the ER and to its intensive degradation by the ubiquitin-proteasomal machinery. However, this is not because of a lack of oligomerization, as demonstrated by the presence of MUT V3R homodimers in the ER. Treatment with SSR149415 restores expression of the mutated receptor on the cell surface and its correct maturation, resulting into the functional recovery of its signaling properties. SSR149415 acts by stabilizing a native-like conformation of the V3R, reducing its association with calnexin and, thus, favoring a secretory pathway rather than the proteasomal degradation pathway. In conclusion, the FN(X)2LL(X)3L sequence is an important motif for the V3R conformation, and the misfolding resulting from its mutation alters the receptor export but can be reverted by SSR149415. PMID- 16210326 TI - Structural and functional evidence for Bacillus subtilis PaiA as a novel N1 spermidine/spermine acetyltransferase. AB - Bacillus subtilis PaiA has been implicated in the negative control of sporulation as well as production of degradative enzymes. PaiA shares recognizable sequence homology with N-acetyltransferases, including those that can acetylate spermidine/spermine substrates. We have determined the crystal structure of PaiA in complex with CoA at 1.9 A resolution and found that PaiA is a member of the N acetyltransferase superfamily of enzymes. Unexpectedly, we observed the binding of an oxidized CoA dimer in the active site of PaiA, and the structural information suggests the substrates of the enzyme could be linear, positively charged compounds. Our biochemical characterization is also consistent with this possibility, since purified PaiA possesses N1-acetyltransferase activity toward polyamine substrates including spermidine and spermine. Further, conditional overexpression of PaiA in bacteria results in increased acetylation of endogenous spermidine pools. Thus, our structural and biochemical analyses indicate that PaiA is a novel N-acetyltransferase capable of acetylating both spermidine and spermine. In this way, the pai operon may function in regulating intracellular polyamine concentrations and/or binding capabilities. In addition to preventing toxicity due to polyamine excess, this function may also serve to regulate expression of certain bacterial gene products such as those involved in sporulation. PMID- 16210328 TI - Gamma-emitting radionuclides in the shallow marine sediments off the Sindh coast, Arabian Sea. AB - Determination of gamma emitting radionuclides in shallow marine sediments off the Sindh coast has been carried out using a gamma spectrometry technique. The activity concentration measured in various sediment samples off the Sindh coast has been found to vary from 15.93 +/- 5.22 to 30.53 +/- 4.70 Bq kg(-1) for 226Ra, from 11.72 +/- 1.22 to 33.94 +/- 1.86 Bq kg(-1) for 228Ra and from 295.22 +/- 32.83 to 748.47 +/- 28.75 Bq kg(-1) for 40K. The calculated mean values of radium equivalent activity, absorbed dose rate and effective dose are 98 Bq kg(-1), 49 nGy h(-1) and 0.06 mSv y(-1), respectively. No artificial radionuclide was detected in the samples measured from the study area. As no data on radioactivity of the coastal environment of Pakistan are available, the data presented here will serve as baseline information on radionuclide concentration in shallow sea sediments off the Sindh coast. The data will also be useful for tracking pollution inventories from unusual radiological events (if any) in the territorial waters of the study area. Further, the information presented will contribute to modelling of a regional radioactivity database from the perspectives of the International Atomic Energy Agency's Asia-Pacific Marine Radioactivity Database and Global Marine Radioactivity Database. PMID- 16210327 TI - The highly abundant protein Ag-lbp55 from Ascaridia galli represents a novel type of lipid-binding proteins. AB - Lipid-binding proteins exhibit important functions in lipid transport, cellular signaling, gene transcription, and cytoprotection. Their functional analogues in nematodes are nematode polyprotein allergens/antigens and fatty acid and retinoid binding proteins. This work describes a novel 55-kDa protein, Ag-lbp55, purified from the parasitic nematode Ascaridia galli. By direct N-terminal sequencing, a partial amino acid sequence was obtained that allowed the design of oligonucleotide primers to obtain the full-length cDNA sequence. Sequence analysis revealed the presence of an N-terminal signal peptide of 25 amino acid residues and a FAR domain at the C terminus. Data base searches showed almost no significant homologies to other described proteins. The secondary structure of Ag lbp55 was predominantly alpha-helical (65%) as shown by CD spectroscopy. It was found to bind with high affinity fatty acids (caprylic, oleic, and palmitic acid) and their fluorescent analogue dansylaminoundecanic acid. Immunolocalization showed that Ag-lbp55 is a highly abundant protein, mainly distributed in the inner hypodermis and extracellularly in the pseudocoelomatic fluid. A similar staining pattern was observed in other pathogenic nematodes, indicating the existence of similar proteins in these species. PMID- 16210329 TI - Dose build up correction for radiation monitors in high-energy bremsstrahlung photon radiation fields. AB - Conventional radiation monitors have been found to underestimate the personal dose equivalent in the high-energy bremsstrahlung photon radiation fields encountered near electron storage rings. Depth-dose measurements in a water phantom were carried out with a radiation survey meter in the bremsstrahlung photon radiation fields from a 450 MeV electron storage ring to find out the magnitude of the underestimation. Dose equivalent indicated by the survey meter was found to build up with increase in thickness of water placed in front of the meter up to certain depth and then reduce with further increase in thickness. A dose equivalent build up factor was estimated from the measurements. An absorbed dose build up factor in a water phantom was also estimated from calculations performed using the Monte Carlo codes, EGS-4 and EGSnrc. The calculations are found to be in very good agreement with the measurements. The studies indicate inadequacy of commercially available radiation monitors for radiation monitoring within shielded enclosures and in streaming high-energy photon radiation fields from electron storage rings, and the need for proper correction for use in such radiation fields. PMID- 16210330 TI - Histone acetylation regulates the cell type specific CIITA promoters, MHC class II expression and antigen presentation in tumor cells. AB - The regulation of MHC class II expression by the class II transactivator (CIITA) is complex and differs in various cell types depending on the relative activity of three CIITA promoters. Here we show that, in plasma cell tumors, the deacetylase inhibitor trichostatin A (TSA) elicits PIII-CIITA but does not activate the IFN-gamma-inducible PIV-CIITA promoter. In trophoblast cells, all CIITA promoter types are constitutively silent and not induced by IFN-gamma or TSA treatment. TSA induction of PI-CIITA was restricted to macrophage and dendritic cell lines. In the Colon 26 tumor IFN-gamma induced endogenous PIV CIITA but not PIII-CIITA while TSA activated class II in the apparent absence of CIITA. Reporter assays in Colon 26 showed that TSA induced PIII-CIITA but not PIV CIITA. Transfection of a dominant negative CIITA plasmid in Colon 26 inhibited induction of class II by IFN-gamma but not TSA. Thus, the potential for both CIITA-dependent and -independent pathways of MHC induction exists within a single cell. Further evidence of CIITA-independent class II expression elicited by TSA was obtained using knockout mice with defects in CIITA, STAT-1alpha and IRF-1 expression. TSA treatment can also activate class II expression in mutant cell lines with deficiencies in signaling molecules, transcription factors and the BRG 1 cofactor that are required for IFN-gamma-induced CIITA expression. Importantly, after epigenetic activation by the deacetylase inhibitor, MHC class II is transported and displayed on the cell surface of a plasma cell tumor and it is converted to an efficient antigen presenting cell for protein and class II peptide presentation. PMID- 16210331 TI - Lung myofibroblasts as targets of salmeterol and fluticasone propionate: inhibition of alpha-SMA and NF-kappaB. AB - Lung myofibroblasts play a major role in the pathophysiology of asthma, contributing not only to tissue remodelling but also to airway inflammation. Nevertheless, only recently, attention has been focused on these cells as potential targets for anti-allergic drugs. Herein, we analysed the pharmacological response of lung myofibroblasts to beta2-agonists associated or not to inhaled corticosteroids, investigating their effects on (i) the constitutive and transforming growth factor-beta (TGF-beta)-induced expression of alpha-smooth muscle actin (alpha-SMA), the main functional marker of myofibroblastic differentiation and contractility; (ii) isometric contraction and (iii) tumour necrosis factor-alpha (TNF-alpha)-induced nuclear translocation of the pro-inflammatory transcription factor nuclear factor-kappaB (NF-kappaB). The beta2-agonist salmeterol (SMl) has on human lung myofibroblasts new direct anti contractile/anti-inflammatory effects that are amplified by the combined use of low concentrations of the glucocorticoid fluticasone propionate (FP). First, SMl and/or FP (10(-12) M) inhibits the constitutive and TGF-beta-induced expression of alpha-SMA. Second, the two drugs block the TNF-alpha-induced nuclear translocation of the pro-inflammatory transcription factor NF-kappaB. Finally, SMl decreases TNF- alpha-induced production of the inflammatory cytokine IL-6. The complementary anti-inflammatory/ anti-contractile effects displayed by SMl and FP on lung myofibroblasts in vitro may be related to the improvement in lung function and symptom control obtained in vivo by the early use of low doses of glucocorticoids in combination with long-acting beta2-agonists. PMID- 16210333 TI - Risk of monoclonal gammopathy of undetermined significance (MGUS) and subsequent multiple myeloma among African American and white veterans in the United States. AB - The age-adjusted incidence of multiple myeloma (MM) is 2-fold higher in African Americans than in whites. A few small studies have reported a higher prevalence of monoclonal gammopathy of undetermined significance (MGUS) in African Americans versus whites. Etiologic factors for MGUS and determinants for transformation of MGUS to MM are unknown. We quantified the prevalence of MGUS and subsequent risk of MM among 4 million African American and white male veterans admitted to Veterans Affairs (VA) hospitals. The age-adjusted prevalence ratio of MGUS in African Americans compared with whites was 3.0 (2.7-3.3 95% confidence interval). Among 2046 MGUS cases, the estimated cumulative risk of MM during the first 10 years of follow-up was similar (P = .37) for African Americans (17%) and whites (15%). In the largest study to date, we suggest that the excess risk of MM in African Americans results from an increase in risk of MGUS rather than an increased risk of progression from MGUS to MM. PMID- 16210332 TI - Severe malarial anemia of low parasite burden in rodent models results from accelerated clearance of uninfected erythrocytes. AB - Severe malarial anemia (SMA) is the most frequent life-threatening complication of malaria and may contribute to the majority of malarial deaths worldwide. To explore the mechanisms of pathogenesis, we developed a novel murine model of SMA in which parasitemias peaked around 1.0% of circulating red blood cells (RBCs) and yet hemoglobin levels fell to 47% to 56% of baseline. The severity of anemia was independent of the level of peak or cumulative parasitemia, but was linked kinetically to the duration of patent infection. In vivo biotinylation analysis of the circulating blood compartment revealed that anemia arose from accelerated RBC turnover. Labeled RBCs were reduced to 1% of circulating cells by 8 days after labeling, indicating that the entire blood compartment had been turned over in approximately one week. The survival rate of freshly transfused RBCs was also markedly reduced in SMA animals, but was not altered when RBCs from SMA donors were transferred into naive recipients, suggesting few functional modifications to target RBCs. Anemia was significantly alleviated by depletion of either phagocytic cells or CD4+ T lymphocytes. This study demonstrates that immunologic mechanisms may contribute to SMA by promoting the accelerated turnover of uninfected RBCs. PMID- 16210334 TI - Sequential heart and autologous stem cell transplantation for systemic AL amyloidosis. AB - Extensive cardiac amyloid deposition in systemic AL amyloidosis is associated with a grave prognosis. Heart transplantation is rarely performed because of the systemic and progressive nature of the disease. Patients with severe cardiac amyloid infiltration are ineligible for the preferred treatment of melphalan chemotherapy with stem cell transplantation (SCT) rescue because of the high risk for treatment-related mortality. Heart transplantation followed by SCT was performed in 5 patients with AL amyloidosis and predominant cardiomyopathy. Patients were followed up for a median of 95 months (range, 37-118 months) from diagnosis. At censor, 3 of 5 patients were well without evidence of intracardiac or extracardiac amyloid accumulation, and median overall survival by Kaplan-Meier estimate was not reached. Two patients died of progressive amyloidosis 33 and 90 months after heart transplantation after relapse of their underlying plasma cell dyscrasia. Heart transplantation followed by SCT is feasible in selected patients with cardiac AL amyloidosis and may confer substantial survival benefit. PMID- 16210335 TI - L-arginine import via cationic amino acid transporter CAT1 is essential for both differentiation and proliferation of erythrocytes. AB - In the present study, we examined the role in hematopoiesis of cationic amino acid transporter 1 (CAT1), which transports L-arginine, L-lysine, L-ornithine, and L-histidine. The expression level of human CAT1 (hCAT1) mRNA in mononuclear cells (MNCs) fractionated according to lineage-selective markers was examined by reverse transcriptase-polymerase chain reaction. The expression of CAT1 in glycophorin A-positive erythroid cells was 8 times higher than in nonfractionated MNC (control) cells. Characteristics of L-arginine uptake by K562 cells, an established leukemic cell line used as an erythroid model, were similar to those of CAT1 in regards to saturation kinetics, sodium independence, and substantial inhibition of L-arginine uptake by N-ethylmaleimide, which is a specific inhibitor of system y+ amino acid transporter. Removal of L-arginine from the culture medium prevented both proliferation and differentiation of K562 cells, while removal of L-lysine or L-histidine had little effect on differentiation, though proliferation was blocked. Hematopoietic stem cells obtained from human cord blood failed to develop into erythroid cells in the absence of L-arginine in the culture medium. These findings indicate that hCAT1 is involved in erythroid hematopoiesis through its role in importing L-arginine, which appears to be essential for the differentiation of red blood cells. PMID- 16210336 TI - Rapamycin, and not cyclosporin A, preserves the highly suppressive CD27+ subset of human CD4+CD25+ regulatory T cells. AB - The immunosuppressive drugs rapamycin and cyclosporin A (CsA) are widely used to prevent allograft rejection. Moreover, they were shown to be instrumental in experimental models of tolerance induction. However, it remains to be elucidated whether these drugs have an effect on the CD4+ CD25+ regulatory T-cell (T(REG)) population, which plays an important role in allograft tolerance. Recently, we reported that alloantigen-driven expansion of human CD4+ CD25+ T(REG)s gives rise to a distinct highly suppressive CD27+ T(REG) subset next to a moderately suppressive CD27- T(REG) subset. In the current study we found that rapamycin and CsA do not interfere with the suppressive activity of human naturally occurring CD4+ CD25+ T cells. However, in contrast to CsA, rapamycin preserved the dominance of the potent CD27+ T(REG) subset over the CD27- T(REG) subset after alloantigen-driven expansion of CD4+ CD25+ T(REG)s in vitro. Accordingly, CD4+ CD25+ T(REG)s cultured in the presence of rapamycin displayed much stronger suppressive capacity than CD4+ CD25+ T(REG)s cultured in the presence of CsA. In addition, CD4+ CD25+ T(REG) cells cultured in the presence of rapamycin, but not CsA, were able to suppress ongoing alloimmune responses. This differential effect of rapamycin and CsA on the CD27+ T(REG) subset dominance may favor the use of rapamycin in tolerance-inducing strategies. PMID- 16210337 TI - Modulation of STAT1 protein levels: a mechanism shaping CD8 T-cell responses in vivo. AB - Type 1 interferons (IFNs) are induced in vivo, administered therapeutically, and potential targets for amelioration of autoimmune diseases. The cytokines mediate profound antiproliferative effects. Signal transducer and activator of transcription 1 (STAT1)-dependent signaling pathways are required for inhibition of proliferation, and viral infections can elicit high levels of type 1 IFNs as well as total STAT1 protein expression. Thus, a mechanism must be in place to help antigen-specific T cells overcome IFN-induced inhibition of proliferation. The studies reported here demonstrate that total CD8 T-cell proliferation in the presence of IFNs, ex vivo in response to cytokines and in vivo during viral infection, is inhibited through a STAT1-dependent mechanism. In contrast, major proportions of antigen-specific CD8, but not CD4, T cells are rendered less sensitive to this inhibition, express lower endogenous levels of total STAT1, and are selectively proliferating in the presence of type 1 IFN, at key times after viral challenge. Taken together, these novel results show that differential STAT1 expression is used by the immune system to modify cytokine-mediated effects on T cell expansion and have implications for the consequences of therapeutic intervention in cytokine function. PMID- 16210338 TI - A novel bispecific protein (ULBP2-BB4) targeting the NKG2D receptor on natural killer (NK) cells and CD138 activates NK cells and has potent antitumor activity against human multiple myeloma in vitro and in vivo. AB - The inability of the immune system to recognize and kill malignant plasma cells in patients with multiple myeloma (MM) has been attributed in part to the ineffective activation of natural killer (NK) cells. In order to activate and target NK cells to the malignant cells in MM we designed a novel recombinant bispecific protein (ULBP2-BB4). While ULBP2 binds the activating NK receptor NKG2D, the BB4 moiety binds to CD138, which is overexpressed on a variety of malignancies, including MM. ULBP2-BB4 strongly activated primary NK cells as demonstrated by a significant increase in interferon-gamma (IFN-gamma) secretion. In vitro, ULBP2-BB4 enhanced the NK-mediated lysis of 2 CD138+ human MM cell lines, U-266 and RPMI-8226, and of primary malignant plasma cells in the allogenic and autologous setting. Moreover, in a nude mouse model with subcutaneously growing RPMI-8226 cells, the cotherapy with ULBP-BB4 and human peripheral blood lymphocytes abrogated the tumor growth. These data suggest potential clinical use of this novel construct in patients with MM. The use of recombinant NK receptor ligands that target NK cells to tumor cells might offer new approaches for other malignancies provided a tumor antigen-specific antibody is available. PMID- 16210339 TI - Developmental and functional significance of the CSF-1 proteoglycan chondroitin sulfate chain. AB - The primary macrophage growth factor, colony-stimulating factor-1 (CSF-1), is homodimeric and exists in 3 biologically active isoforms: a membrane-spanning, cell-surface glycoprotein (csCSF-1) and secreted glycoprotein (sgCSF-1) and proteoglycan (spCSF-1) isoforms. To investigate the in vivo role of the chondroitin sulfate glycosaminoglycan (GAG) chain of spCSF-1, we created mice that exclusively express, in a normal tissue-specific and developmental manner, either the secreted precursor of spCSF-1 or the corresponding precursor in which the GAG addition site was mutated. The reproductive, hematopoietic tooth eruption and tissue macrophage defects of CSF-1-deficient, osteopetrotic Csf1(op)/Csf1(op) mice were corrected by transgenic expression of the precursors of either sgCSF-1 or spCSF-1. Furthermore, in contrast to the transgene encoding csCSF-1, both failed to completely correct growth retardation, suggesting a role for csCSF-1 in the regulation of body weight. However, spCSF-1, in contrast to sgCSF-1, completely resolved the osteopetrotic phenotype. Furthermore, in transgenic lines expressing different concentrations of sgCSF-1 or spCSF-1, spCSF-1 more efficiently corrected Csf1(op)/Csf1(op) defects of tooth eruption, eyelid opening, macrophage morphology, and B-cell deficiency than sgCSF-1. These results indicate an important role of the CSF-1 chondroitin sulfate proteoglycan in in vivo signaling by secreted CSF-1. PMID- 16210340 TI - Antibody-induced neutrophil activation as a trigger for transfusion-related acute lung injury in an ex vivo rat lung model. AB - Transfusion-related acute lung injury (TRALI) is a hazardous complication of transfusion and has become the leading cause of transfusion-related death in the United States and United Kingdom. Although leukoagglutinating antibodies have been frequently shown to be associated with the syndrome, the mechanism by which they induce TRALI is poorly understood. Therefore, we reproduced TRALI in an ex vivo rat lung model. Our data demonstrate that TRALI induction by antileukocyte antibodies is dependent on the density of the cognate antigen but does not necessarily require leukoagglutinating properties of the antibody or the presence of complement proteins. Rather, antibody-mediated activation of neutrophils seems to initiate TRALI, a process that could be triggered by neutrophil stimulation with fMLP. Antibody-mediated neutrophil activation and subsequent release of reactive oxygen species may thus represent key events in the pathophysiologic cascade that leads to immune TRALI. PMID- 16210342 TI - T-cell non-Hodgkin lymphoma. AB - T-cell non-Hodgkin lymphomas (NHLs) are uncommon malignancies. The current WHO/EORTC classification recognizes 9 distinct clinicopathologic peripheral T cell NHLs. These disorders have unique characteristics and require individualized diagnostic and therapeutic strategies. Tremendous progress has been made in recent years in the understanding of the pathogenesis of these disorders. Specific chromosomal translocations and viral infections are now known to be associated with certain lymphomas. In this review, we describe their clinical and pathologic features. We also discuss the use of molecular studies in the diagnostic work-up of T-cell lymphomas. Because of the rarity of these disorders and the lack of well-designed clinical trials, the treatment of peripheral T-cell NHLs is often challenging. Additional studies are required to learn more about the biology of these diseases, which may lead to more optimal and possibly targeted therapies. PMID- 16210341 TI - Sequential activation of p38 and ERK pathways by cGMP-dependent protein kinase leading to activation of the platelet integrin alphaIIb beta3. AB - Integrin activation (inside-out signaling) in platelets can be initiated by agonists such as von Willebrand factor (VWF) and thrombin. Here we show that a mitogen-activated protein kinase (MAPK), p38, plays an important role in the activation of integrin alphaIIb beta3 induced by VWF and thrombin. A dominant negative mutant of p38, p38AF, inhibits alphaIIb beta3 activation induced by VWF binding to its receptor, the platelet glycoprotein Ib-IX (GPIb-IX), and p38 inhibitors diminish platelet aggregation induced by VWF or low-dose thrombin. The inhibitory effect of p38 inhibitor is unlikely to be caused by the previous suggested effect on cyclo-oxygenase, as inhibition also was observed in the presence of high concentrations of cyclo-oxygenase inhibitor, aspirin. VWF or thrombin induces p38 activation, which is inhibited in cGMP-dependent protein kinase (PKG)-knockout mouse platelets and PKG inhibitor-treated human platelets, indicating that activation of p38 is downstream from PKG in the signaling pathway. p38AF or p38 inhibitors diminish PKG-induced phosphorylation of extracellular stimuli-responsive kinase (ERK), which also is important in integrin activation. Thus, p38 plays an important role in mediating PKG-dependent activation of ERK. These data delineate a novel signaling pathway in which platelet agonists sequentially activate PKG, p38, and ERK pathways leading to integrin activation. PMID- 16210343 TI - Von Hippel-Lindau-dependent polycythemia is endemic on the island of Ischia: identification of a novel cluster. AB - Chuvash polycythemia (MIM 263400) is an autosomal recessive disorder characterized by a high hemoglobin level, relatively high serum erythropoietin, and early death. It results from a Von Hippel-Lindau (VHL) gene mutation (C598T) that causes increased HIF-1alpha activity and erythrocyte production in the face of normoxia. This polycythemia is endemic in Chuvashia, whereas its worldwide frequency is very low. We investigated the incidence of the Chuvash-type VHL mutation in Campania (South Italy) and identified 14 affected subjects (5 families). Twelve live on the island of Ischia (Bay of Naples). From analysis of the mutated allele, we found that the disease was more frequent on Ischia (0.070) than in Chuvashia (0.057). The haplotype of all patients matched that identified in the Chuvash cluster, thereby supporting the single-founder hypothesis. We also found that nonaffected heterozygotes had increased HIF-1alpha activity, which might confer a biochemical advantage for mutation maintenance. In conclusion, we have identified the first large cluster of Chuvash erythrocytosis outside Chuvashia, which suggests that this familial polycythemia might be endemic in other regions of the world. PMID- 16210344 TI - Cyclic guanosine 5'-monophosphate-dependent protein kinase II is induced by luteinizing hormone and progesterone receptor-dependent mechanisms in granulosa cells and cumulus oocyte complexes of ovulating follicles. AB - Cyclic GMP (cGMP)-dependent protein kinase II (Prkg2, cGK II) was identified as a potential target of the progesterone receptor (Nr3c3) in the mouse ovary based on microarray analyses. To document this further, the expression patterns of cGK II and other components of the cGMP signaling pathway were analyzed during follicular development and ovulation using the pregnant mare serum gonadotropin (PMSG)-human chorionic gonadotropin (hCG)-primed immature mice. Levels of cGK II mRNA were low in ovaries of immature mice, increased 4-fold in response to pregnant mare serum gonadotropin and 5-fold more within 12 h after hCG, the time of ovulation. In situ hybridization localized cGK II mRNA to granulosa cells and cumulus oocyte complexes of periovulatory follicles. In progesterone receptor (PR) null mice, cGK II mRNA was reduced significantly at 12 h after hCG in contrast to heterozygous littermates. In primary granulosa cell cultures, cGK II mRNA was induced by phorbol 12-myristate 13-acetate enhanced by adenoviral expression of PR-A and blocked by RU486 and trilostane. PR-A in the absence of phorbol 12-myristate 13-acetate was insufficient to induce cGK II. Expression of cGK I (Prkg1) was restricted to the residual tissue and not regulated by hormones. Guanylate cyclase-A (Npr1; GC-A) mRNA expression increased 6-fold by 4 h after hCG treatment in contrast to pregnant mare serum gonadotropin alone and was localized to granulosa cells of preovulatory follicles. Collectively, these data show for the first time that cGK II (not cGK I) and GC-A are selectively induced in granulosa cells of preovulatory follicles by LH- and PR-dependent mechanisms, thereby providing a pathway for cGMP function during ovulation. PMID- 16210345 TI - A glycoprotein hormone expressed in corticotrophs exhibits unique binding properties on thyroid-stimulating hormone receptor. AB - Corticotroph-derived glycoprotein hormone (CGH), also referred to as thyrostimulin, is a noncovalent heterodimer of glycoprotein hormone alpha 2 (GPHA2) and glycoprotein hormone beta 5 (GPHB5). Here, we demonstrate that both subunits of CGH are expressed in the corticotroph cells of the human anterior pituitary, as well as in skin, retina, and testis. CGH activates the TSH receptor (TSHR); (125)I-CGH binding to cells expressing TSHR is saturable, specific, and of high affinity. In competition studies, unlabeled CGH is a potent competitor for (125)I-TSH binding, whereas unlabeled TSH does not compete for (125)I-CGH binding. Binding and competition analyses are consistent with the presence of two binding sites on the TSHR transfected baby hamster kidney cells, one that can interact with either TSH or CGH, and another that binds CGH alone. Transgenic overexpression of GPHB5 in mice produces elevations in serum T(4) levels, reductions in body weight, and proptosis. However, neither transgenic overexpression of GPHA2 nor deletion of GPHB5 produces an overt phenotype in mice. In vivo administration of CGH to mice produces a dose-dependent hyperthyroid phenotype including elevation of T(4) and hypertrophy of cells within the inner adrenal cortex. However, the distinctive expression patterns and binding characteristics of CGH suggest that it has endogenous biological roles that are discrete from those of TSH. PMID- 16210346 TI - Why muscle stops building when it's working. PMID- 16210347 TI - A role for leucine in rejuvenating the anabolic effects of food in old rats. PMID- 16210349 TI - Spontaneous activity of lower urinary tract smooth muscles: correlation between ion channels and tissue function. AB - Smooth muscles from the urethra and bladder display characteristic patterns of spontaneous contractile activity in the filling phase of the micturition cycle. Tonic contractions are seen in the urethral smooth muscles, and phasic contractions occur in the detrusor. Overactivity in the detrusor is a common clinical problem. The ion channels in the smooth muscle membranes play an important role in determining the functional properties, and are obvious targets for treatment of the overactive bladder. Recent evidence suggests that interstitial cells may also play a role in determining the pattern of spontaneous activity, although their precise role is less well established in the urinary tract than in the gut. The ion channels involved in these cells are also of interest. This review discusses what is known of ion channels in these tissues, and their implications for function. PMID- 16210348 TI - Kv1 channels selectively prevent dendritic hyperexcitability in rat Purkinje cells. AB - Purkinje cells, the sole output of the cerebellar cortex, encode the timing signals required for motor coordination in their firing rate and activity pattern. Dendrites of Purkinje cells express a high density of P/Q-type voltage gated calcium channels and fire dendritic calcium spikes. Here we show that dendritic subthreshold Kv1.2 subunit-containing Kv1 potassium channels prevent generation of random spontaneous calcium spikes. With Kv1 channels blocked, dendritic calcium spikes drive bursts of somatic sodium spikes and prevent the cell from faithfully encoding motor timing signals. The selective dendritic function of Kv1 channels in Purkinje cells allows them to effectively suppress dendritic hyperexcitability without hindering the generation of somatic action potentials. Further, we show that Kv1 channels also contribute to dendritic integration of parallel fibre synaptic input. Kv1 channels are often targeted to soma and axon and the data presented support a major dendritic function for these channels. PMID- 16210351 TI - Exercise rapidly increases eukaryotic elongation factor 2 phosphorylation in skeletal muscle of men. AB - Protein synthesis in skeletal muscle is known to decrease during contractions but the underlying regulatory mechanisms are unknown. Here, the effect of exercise on skeletal muscle eukaryotic elongation factor 2 (eEF2) phosphorylation, a key component in protein translation machinery, was examined. Eight healthy men exercised on a cycle ergometer at a workload eliciting approximately 67% peak pulmonary oxygen consumption (VO2 peak) with skeletal muscle biopsies taken from the vastus lateralis muscle at rest as well as after 1, 10, 30, 60 and 90 min of exercise. In response to exercise, there was a rapid (i.e. < 1 min) 5- to 7-fold increase in eEF2 phosphorylation at Thr56 that was sustained for 90 min of continuous exercise. The in vitro activity of skeletal muscle eEF2 kinase was not altered by exercise indicating that the increased activity of eEF2 kinase to eEF2 is not mediated by covalent mechanisms. In support of this, the increase in AMPK activity was temporally unrelated to eEF2 phosphorylation. However, skeletal muscle eEF2 kinase was potently activated by Ca(2)(+)-calmodulin in vitro, suggesting that the higher eEF2 phosphorylation in working skeletal muscle is mediated by allosteric activation of eEF2 kinase by Ca(2)(+) signalling via calmodulin. Given that eEF2 phosphorylation inhibits eEF2 activity and mRNA translation, these findings suggest that the inhibition of protein synthesis in contracting skeletal muscle is due to the Ca(2)(+)-induced stimulation of eEF2 kinase. PMID- 16210350 TI - Different neuronal populations of the rat median preoptic nucleus express c-fos during sleep and in response to hypertonic saline or angiotensin-II. AB - The median preoptic nucleus (MnPN) of the hypothalamus contains sleep-active neurones, and sleep-related Fos-immunoreactivity (IR) in this nucleus is primarily expressed in GABAergic cells. The MnPN also contains cells responsive to hypertonic saline and to angiotensin-II (Ang-II). To clarify functional relationships between MnPN neurones involved in the regulation of sleep and body fluid homeostasis, we examined c-fos expression in the MnPN after administration of hypertonic saline and Ang-II in both spontaneously sleeping and sleep-deprived rats. Systemic administration of hypertonic saline and intracerebroventricular (i.c.v.) injection of Ang-II increased Fos-IR in both spontaneously sleeping and sleep-deprived rats, compared to control animals. To determine if the population of MnPN neurones activated in response to osmotic and hormonal stimuli is similar to or different from neurones activated during sleep, we quantified Fos-IR in MnPN GABAergic neurones in spontaneously sleeping hypertonic saline- and Ang-II treated rats versus respective control rats. Fos-IR evoked by these treatments occurred primarily (80-85%) in non-GABAergic neurones. Findings of the present study provide evidence that separate populations of MnPN neurones are involved in the regulation of sleep and body fluid homeostasis. PMID- 16210352 TI - Differential targeting and functional specialization of sodium channels in cultured cerebellar granule cells. AB - The ion channel dynamics that underlie the complex firing patterns of cerebellar granule (CG) cells are still largely unknown. Here, we have characterized the subcellular localization and functional properties of Na+ channels that regulate the excitability of CG cells in culture. As evidenced by RT-PCR and immunocytochemical analysis, morphologically differentiated CG cells expressed Nav1.2 and Nav1.6, though both subunits appeared to be differentially regulated. Nav1.2 was localized at most axon initial segments (AIS) of CG cells from 8 days in vitro DIV 8 to DIV 15. At DIV 8, Nav1.6 was found uniformly throughout somata, dendrites and axons with occasional clustering in a subset of AIS. Accumulation of Nav1.6 at most AIS was evident by DIV 13-14, suggesting it is developmentally regulated at AIS. The specific contribution of these differentially distributed Na+ channels has been assessed using a combination of methods that allowed discrimination between functionally compartmentalized Na+ currents. In agreement with immunolocalization, we found that fast activating-fully inactivating Na+ currents predominate at the AIS membrane and in the somatic plasma membrane. PMID- 16210353 TI - Effect of hydration on interstitial distribution of charged albumin in rat dermis in vitro. AB - At physiological pH, negatively charged glycosaminoglycans in the extracellular matrix may influence distribution volume of macromolecular probes, a phenomenon of importance for hydration of the interstitium and therefore for body fluid balance. We hypothesized that such charge effect was dependent on hydration. Human serum albumin (HSA) (the pH value for the isoelectric point (pI) = 4.9) was made neutral by cationization (cHSA) (pI = 7.6). Rat dermis was studied in vitro in a specially designed equilibration cell allowing control of hydration. Using a buffer containing labelled native HSA and cHSA, the distribution volumes were calculated relative to that of 51Cr-EDTA, an extracellular tracer. During changes in hydration (H), defined as (wet weight - dry weight) (dry weight)(-1)), the slope of the equation describing the relationship between extracellular fluid volume (V(x)) (in g H2O (g dry weight)(-1)) and H (V(x) = 0.925 H + 0.105) differed significantly from that for available volumes of cHSA (V(a,cHSA) = 0.624 H - 0.538) and HSA (V(a,HSA) = 0.518 H - 0.518). A gradual reduction in H led to a reduction in difference between available volumes for the two albumin species. Screening the fixed charges by 1 m NaCl resulted in similar available and excluded volumes of native HSA and neutral cHSA. We conclude that during gradual dehydration, there is a reduced effect of fixed negative charges on interstitial exclusion of charged macromolecules. This effect may be explained by a reduced hydration domain surrounding tissue and probe macromolecules in conditions of increased electrostatic interactions. Furthermore, screening of negative charges suggested that hyaluronan associated with collagen may influence intrafibrillar volume of collagen and thereby available and excluded volume fraction. PMID- 16210355 TI - The effect of changes in cardiac output on middle cerebral artery mean blood velocity at rest and during exercise. AB - We examined the relationship between changes in cardiac output and middle cerebral artery mean blood velocity (MCA V(mean)) in seven healthy volunteer men at rest and during 50% maximal oxygen uptake steady-state submaximal cycling exercise. Reductions in were accomplished using lower body negative pressure (LBNP), while increases in were accomplished using infusions of 25% human serum albumin. Heart rate (HR), arterial blood pressure and MCA V(mean) were continuously recorded. At each stage of LBNP and albumin infusion was measured using an acetylene rebreathing technique. Arterial blood samples were analysed for partial pressure of carbon dioxide tension (P(a,CO2). During exercise HR and were increased above rest (P < 0.001), while neither MCA V(mean) nor P(a,CO2) was altered (P > 0.05). The MCA V(mean) and were linearly related at rest (P < 0.001) and during exercise (P = 0.035). The slope of the regression relationship between MCA V(mean) and at rest was greater (P = 0.035) than during exercise. In addition, the phase and gain between MCA V(mean) and mean arterial pressure in the low frequency range were not altered from rest to exercise indicating that the cerebral autoregulation was maintained. These data suggest that the associated with the changes in central blood volume influence the MCA V(mean) at rest and during exercise and its regulation is independent of cerebral autoregulation. It appears that the exercise induced sympathoexcitation and the change in the distribution of between the cerebral and the systemic circulation modifies the relationship between MCA V(mean) and . PMID- 16210354 TI - Failure of cAMP agonists to activate rescued deltaF508 CFTR in CFBE41o- airway epithelial monolayers. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a cyclic AMP regulated chloride channel. Mutations in the CFTR gene result in cystic fibrosis (CF). The most common mutation, deltaF508, results in endoplasmic reticulum associated degradation (ERAD) of CFTR. DeltaF508 CFTR has been described as a temperature-sensitive mutation that can be rescued following growth at 27 degrees C. In order to study the processing and function of wild-type and rescued deltaF508 CFTR at the cell surface under non-polarized and polarized conditions, we developed stable cell lines expressing deltaF508 or wild-type CFTR. CFBE41o- is a human airway epithelial cell line capable of forming high resistance, polarized monolayers when cultured on permeable supports, while HeLa cells are normally grown under non-polarizing conditions. Immunoprecipitation, cell surface biotinylation, immunofluorescence, and functional assays confirmed the presence of deltaF508 CFTR at the cell surface in both cell lines after incubating the cells for 48 h at 27 degrees C. However, stimulators of wild-type CFTR such as forskolin, beta2-adrenergic or A2B-adenosine receptor agonists failed to activate rescued deltaF508 CFTR in CFBE41o- monolayers. Rescued deltaF508 CFTR could be stimulated with genistein independent of pretreatment with cAMP signalling agonists. Interestingly, rescued deltaF508 CFTR in HeLa cells could be efficiently stimulated with either forskolin or genistein to promote Cl- transport. These results indicate that deltaF508 CFTR, when rescued in CFBE41o- human airway epithelial cells, is poorly responsive to signalling pathways known to regulate wild-type CFTR. Furthermore, the differences in rescue and activation of deltaF508 CFTR in the two cell lines suggest that cell-type specific differences in deltaF508 CFTR processing are likely to complicate efforts to identify potentiators and/or correctors of the deltaF508 defect. PMID- 16210356 TI - Actions of brain-derived neurotrophic factor on spinal nociceptive transmission during inflammation in the rat. AB - The aim of the current study was to investigate whether, and if so how, brain derived neurotrophic factor (BDNF) acts to develop the spinal sensitization underlying inflammation-induced hyperalgesia. In spinal cord slice preparations from rats with inflammation induced by complete Freund's adjuvant (CFA), BDNF, but not nerve growth factor (NGF) or neurotrophin-3 (NT-3), acted presynaptically to increase the frequency of excitatory miniature EPSCs in substantia gelatinosa (SG) neurones of the CFA-treated, but not untreated rats, through activation of lidocaine (lignocaine)-sensitive, TTX-resistant Na+ channels. This effect was observed in the spinal cord slices of the CFA-treated rat only 2-4 days after the CFA injection. On the other hand, the number of monosynaptic Abeta afferent inputs to the SG significantly increased 1 week after the onset of the inflammation, and this increase was significantly suppressed by treatment with anti-BDNF antiserum administered 1 day before and just after the CFA injection. In addition, the treatment with anti-BDNF antiserum significantly attenuated the CFA-induced hyperalgesia and/or allodynia. These findings, taken together, suggest that BDNF, which is considered to be released from the sensitized primary afferents, increases the excitability of SG neurones through its action on the presynaptic terminals. BDNF may thereafter induce monosynaptic Abeta afferents to the SG, thereby developing hyperalgesia and/or allodynia during inflammation. PMID- 16210357 TI - Vertebral position alters paraspinal muscle spindle responsiveness in the feline spine: effect of positioning duration. AB - Proprioceptive information from paraspinal tissues including muscle contributes to neuromuscular control of the vertebral column. We investigated whether the history of a vertebra's position can affect signalling from paraspinal muscle spindles. Single unit recordings were obtained from muscle spindle afferents in the L6 dorsal roots of 30 Nembutal-anaesthetized cats. Each afferent's receptive field was in the intact muscles of the low back. The L6 vertebra was controlled using a displacement-controlled feedback motor and was held in each of three different conditioning positions for durations of 0, 2, 4, 6 and 8 s. Conditioning positions (1.0-2.2 mm dorsal and ventral relative to an intermediate position) were based upon the displacement that loaded the L6 vertebra to 50-60% of the cat's body weight. Following conditioning positions that stretched (hold long) and shortened (hold-short) the spindle, the vertebra was repositioned identically and muscle spindle discharge at rest and to movement was compared with conditioning at the intermediate position. Hold-short conditioning augmented mean resting spindle discharge by +4.1 to +6.2 impulses s(-1); however, the duration of hold-short did not significantly affect this increase (F(4,145) = 0.49, P = 0.74). The increase was maintained at the beginning of vertebral movement but quickly returned to baseline. Conversely, hold-long conditioning significantly diminished mean resting spindle discharge by -2.0 to -16.1 impulses s(-1) (F(4,145) = 11.23, P < 0.001). The relationship between conditioning duration and the diminished resting discharge could be described by a quadratic (F(1,145) = 9.28, P = 0.003) revealing that the effects of positioning history were fully developed within 2 s of conditioning. In addition, 2 s or greater of hold-long conditioning significantly diminished spindle discharge to vertebral movement by -5.7 to -10.0 impulses s(-1) (F(4,145) = 11.0, P < 0.001). These effects of vertebral positioning history may be a mechanism whereby spinal biomechanics interacts with the spine's proprioceptive system to produce acute effects on neuromuscular control of the vertebral column. PMID- 16210358 TI - Sodium channel dysfunction in intractable childhood epilepsy with generalized tonic-clonic seizures. AB - Mutations in SCN1A, the gene encoding the brain voltage-gated sodium channel alpha(1) subunit (Na(V)1.1), are associated with genetic forms of epilepsy, including generalized epilepsy with febrile seizures plus (GEFS+ type 2), severe myoclonic epilepsy of infancy (SMEI) and related conditions. Several missense SCN1A mutations have been identified in probands affected by the syndrome of intractable childhood epilepsy with generalized tonic-clonic seizures (ICEGTC), which bears similarity to SMEI. To test whether ICEGTC arises from molecular mechanisms similar to those involved in SMEI, we characterized eight ICEGTC missense mutations by whole-cell patch clamp recording of recombinant human SCN1A heterologously expressed in cultured mammalian cells. Two mutations (G979R and T1709I) were non-functional. The remaining alleles (T808S, V983A, N1011I, V1611F, P1632S and F1808L) exhibited measurable sodium current, but had heterogeneous biophysical phenotypes. Mutant channels exhibited lower (V983A, N1011I and F1808L), greater (T808S) or similar (V1611F and P1632S) peak sodium current densities compared with wild-type (WT) SCN1A. Three mutations (V1611F, P1632S and F1808L) displayed hyperpolarized conductance-voltage relationships, while V983A exhibited a strong depolarizing shift in the voltage dependence of activation. All mutants except T808S had hyperpolarized shifts in the voltage dependence of steady-state channel availability. Three mutants (V1611F, P1632S and F1808L) exhibited persistent sodium current ranging from approximately 1-3% of peak current amplitude that was significantly greater than WT-SCN1A. Several mutants had impaired slow inactivation, with V983A showing the most prominent effect. Finally, all of the functional alleles exhibited reduced use-dependent channel inhibition. In summary, SCN1A mutations associated with ICEGTC result in a wide spectrum of biophysical defects, including mild-to-moderate gating impairments, shifted voltage dependence and reduced use dependence. The constellation of biophysical abnormalities for some mutants is distinct from those previously observed for GEFS+ and SMEI, suggesting possible, but complex, genotype-phenotype correlations. PMID- 16210359 TI - Dihydrotestosterone inhibits insulin-stimulated cyclin D2 messenger ribonucleic acid expression in rat ovarian granulosa cells by reducing the phosphorylation of insulin receptor substrate-1. AB - The effect of 5alpha-dihydrotestosterone (DHT) on insulin-stimulated granulosa cell proliferation was examined using cyclin D2 mRNA as a marker. Granulosa cells from 3-d estradiol-treated immature rats showed a concentration-dependent increase in cyclin D2 mRNA expression in response to insulin. Exposure to DHT reduced the insulin-stimulated cyclin D2 mRNA expression. Inhibition of the two insulin-signaling pathways, ERK and phosphatidylinositol 3 kinase (PI3 kinase), by using specific inhibitors, also reduced this insulin-stimulated response. These results suggest that both ERK and PI3 kinase signaling are involved in insulin stimulated granulosa cell proliferation. DHT exposure resulted in reduced insulin-stimulated ERK phosphorylation. DHT treatment also reduced the insulin mediated insulin receptor substrate-1 and Raf-1 phosphorylation, the upstream molecules of ERK in insulin signaling pathway. Additionally, inhibition of insulin stimulated PI3 kinase activation reduced ERK phosphorylation. The present study therefore shows that the inhibitory effect of DHT on insulin-stimulated granulosa cell proliferation occurs early in the signaling pathway at the level of insulin receptor substrate-1 phosphorylation, leading to reduced ERK phosphorylation and subsequent inhibition of cyclin D2 mRNA expression. PMID- 16210360 TI - Stimulation of the novel estrogen receptor-alpha intronic TERP-1 promoter by estrogens, androgen, pituitary adenylate cyclase-activating peptide, and forskolin, and autoregulation by TERP-1 protein. AB - The estrogen receptor-alpha (ERalpha) pituitary-specific variant, TERP-1, is regulated dramatically by physiological status. We examined hormonal regulation of the TERP-1 promoter in transient transfection assays in GH3 somatolactotrope cells. We found that 17beta-estradiol (E2), genistein, androgen, pituitary adenylate cyclase-activating peptide, and forskolin (FSK) all stimulated TERP-1 promoter activity, whereas progesterone had no effect. ERalpha bound to a palindromic estrogen response element (ERE) and two half-site EREs; mutation of any of these sites decreased basal expression and completely obliterated E2 stimulation. In contrast, mutation of an activator protein-1 site decreased basal and FSK-stimulated promoter activity, but not E2 or androgen stimulation. The pure antiestrogen ICI 182,780 suppressed E2 and genistein, but not FSK or androgen, stimulation. Similarly, mutation of the ERE palindrome or half-site EREs suppressed promoter stimulation by E2 and genistein, but not by androgen or FSK. Because TERP-1 levels regulate ERalpha function on model promoters, we tested TERP-1 modulation of its own and other physiological promoters. TERP-1 suppressed basal and E2-stimulated expression of its own promoter. TERP-1 suppression required the ERE regions of the promoter, and the dimerization domain of TERP-1. TERP-1 overexpression also suppressed E2 stimulation of the progesterone receptor and prolactin promoters. Thus, estrogens, androgen, and FSK can stimulate TERP-1 promoter activity, and increased TERP-1 expression modulates E2 stimulation of physiological promoters. These data suggest that TERP-1 regulation may play a significant role in modifying pituitary ERalpha responses. PMID- 16210361 TI - Effects of central infusion of ghrelin on food intake and plasma levels of growth hormone, luteinizing hormone, prolactin, and cortisol secretion in sheep. AB - Ghrelin is an endogenous ligand for the GH secretagogue/ghrelin receptor (GHS-R) and stimulates feeding behavior and GH levels in rodents and humans. A preprandial increase in plasma ghrelin levels is seen in sheep on programmed feeding, followed by a postprandial rise in plasma GH levels, but effects on food intake and endocrine function are not defined in this ruminant species. We administered ghrelin to female sheep in various modes and measured effects on voluntary food intake (VFI) and plasma levels of GH, LH, prolactin, and cortisol. Whether administered intracerebroventricularly or iv, ghrelin consistently failed to stimulate VFI. On the other hand, ghrelin invariably increased plasma GH levels and alpha,beta-diaminopropanoic acid-octanoyl3 human ghrelin was more potent than ovine ghrelin. Bolus injection of ghrelin into the third cerebral ventricle reduced plasma LH levels but did not affect levels of prolactin or cortisol. These findings suggested that the preprandial rise in plasma ghrelin that is seen in sheep on programmed feeding does not influence VFI but is likely to be important in the postprandial rise in GH levels. Thus, ghrelin does not appear to be a significant regulator of ingestive behavior in this species of ruminant but acts centrally to indirectly regulate GH and LH secretion. PMID- 16210362 TI - Corticotropin-releasing hormone directly stimulates thermogenesis in skeletal muscle possibly through substrate cycling between de novo lipogenesis and lipid oxidation. AB - The mechanisms by which CRH and related peptides (i.e. the CRH/urocortin system) exert their control over thermogenesis and weight regulation have until now focused only upon their effects on brain centers controlling sympathetic outflow. Using a method that involves repeated oxygen uptake determinations in intact mouse skeletal muscle, we report here that CRH can act directly on skeletal muscle to stimulate thermogenesis, an effect that is more pronounced in oxidative than in glycolytic muscles and that can be inhibited by a selective CRH-R2 antagonist or blunted by a nonselective CRH receptor antagonist. This thermogenic effect of CRH can also be blocked by interference along pathways of de novo lipogenesis and lipid oxidation, as well as by inhibitors of phosphatidylinositol 3-kinase or AMP-activated protein kinase. Taken together, these studies demonstrate that CRH can directly stimulate thermogenesis in skeletal muscle, and in addition raise the possibility that this thermogenic effect, which requires both phosphatidylinositol 3-kinase and AMP-activated protein kinase signaling, might occur via substrate cycling between de novo lipogenesis and lipid oxidation. The effect of CRH in directly stimulating thermogenesis in skeletal muscle underscores a potentially important peripheral role for the CRH/urocortin system in the control of thermogenesis in this tissue, in its protection against excessive intramyocellular lipid storage, and hence against skeletal muscle lipotoxicity and insulin resistance. PMID- 16210363 TI - T helper type 1 and type 2 cytokines exert divergent influence on the induction of prostaglandin E2 and hyaluronan synthesis by interleukin-1beta in orbital fibroblasts: implications for the pathogenesis of thyroid-associated ophthalmopathy. AB - Thyroid-associated ophthalmopathy (TAO) is an autoimmune component of Graves' disease characterized by intense inflammation in the setting of volume expansion. At the heart of orbital susceptibility to Graves' disease appears to be the peculiar phenotype of orbital fibroblasts that, when activated by IL-1beta and other proinflammatory cytokines, produce excess prostaglandin E2 (PGE2) and hyaluronan. T helper type 1 (Th1) cytokines predominate early in TAO, whereas Th2 cytokines are more abundant later. It is currently unknown whether this transition might promote changes in tissue reactivity associated with disease progression. We report here that interferon-gamma and IL-4, representative of these respective classes of cytokines, attenuate IL-1beta-provoked PGE2 production. This down-regulation is mediated by blocking the induction of prostaglandin endoperoxide H synthase-2 (PGHS-2), the inflammatory cyclooxygenase. The mechanism involves blockade by IL-4 and interferon-gamma of the IL-1beta-dependent activation of PGHS-2 gene promoter activity. In addition, interferon gamma inhibits IL-1beta-provoked PGHS-2 mRNA stability. The actions of interferon-gamma and IL-4 are mediated through the Janus kinase 2/signal transducer and activator of transcription signaling pathway and could be abolished by treating with AG490, a specific inhibitor of Janus kinase 2. In contrast, the up-regulation of hyaluronan synthesis by IL-1beta is enhanced by either IL-4 or interferon-gamma. The latter two cytokines enhance the induction by IL-1beta of hyaluronan synthase-2 expression. These unexpected findings indicate that the Th1-->Th2 cytokine transition exerts equivalent influence on PGE2 and hyaluronan production as TAO progresses from early to late stage. PMID- 16210364 TI - Bone morphogenetic protein-15 in the zebrafish ovary: complementary deoxyribonucleic acid cloning, genomic organization, tissue distribution, and role in oocyte maturation. AB - Bone morphogenetic protein-15 (BMP-15) is a member of the TGFbeta family known to regulate ovarian functions in mammals. The structure and function of BMP-15 in lower vertebrates are less known. In this study, we cloned the zebrafish BMP-15 (zfBMP-15) cDNA and depicted its genomic organization. The zfBMP-15 cDNA encodes a protein of 384 amino acids. The mature protein has 46-51% sequence identities to fugu, chicken, and mammalian BMP-15. It also shares 38-46% homology with growth and differentiation factor-9 in fishes, chicken, and mammals. Phylogenetic analysis further confirms that the zfBMP-15 is most closely related to BMP-15 from other species, whereas the growth and differentiation factor-9 peptides from fish to mammals form a distinct branch. Comparison of zfBMP-15 cDNA with zebrafish genome database revealed that zfBMP-15 is encoded by a gene with two exons and one intron, located on chromosome 6. BMP-15 mRNA is expressed in the ovary and testis and, to a lesser extent, brain, liver, gut, heart, and muscle. Real-time PCR revealed that BMP-15 is expressed in follicles at all stages of development with no significant changes over the course of folliculogenesis. Using in situ hybridization and immunocytochemistry, we detected BMP-15 in both oocytes and follicular cells. Incubation of follicles with antiserum against zfBMP15 increased oocyte maturation, whereas incubation with recombinant human BMP-15 suppressed human chorionic gonadotropin-induced oocyte maturation. These findings suggest that BMP-15 plays a role in regulating gonadal functions in fish, in particular oocyte maturation. PMID- 16210365 TI - Direct, androgen receptor-mediated regulation of the FKBP5 gene via a distal enhancer element. AB - Androgen signaling via the androgen receptor (AR) transcription factor is crucial to normal prostate homeostasis and prostate tumorigenesis. Current models of AR function are predominantly based on studies of prostate-specific antigen regulation in androgen-responsive cell lines. To expand on these in vitro paradigms, we used the mouse prostate to elucidate the mechanisms through which AR regulates another direct target, FKBP5, in vivo. FKBP5 encodes an immunophilin that has been previously implicated in glucocorticoid and progestin signaling pathways and that likely influences prostate physiology in the presence of androgens. In this work, we show that androgens directly regulate FKBP5 via an interaction between the AR and a distal enhancer located 65 kb downstream of the transcription start site in the fifth intron of the FKBP5 gene. We have found that AR selectively recruits cAMP response element-binding protein to this enhancer. These interactions, in turn, result in chromatin remodeling that affects the enhancer proper but not the FKBP5 locus as a whole. Furthermore, in contrast to prostate-specific antigen-regulatory mechanisms, we show that transactivation of the FKBP5 gene does not rely on a single looping complex to mediate communication between the distal enhancer and proximal promoter. Rather, the distal enhancer complex and basal transcription apparatus communicate indirectly with one another, implicating a regulatory mechanism that has not been previously appreciated for AR target genes. PMID- 16210366 TI - Both testosterone and follicle-stimulating hormone independently inhibit spermatogonial differentiation in irradiated rats. AB - Simultaneous suppression of both testosterone and FSH with GnRH antagonists (GnRH ant) reverses the radiation-induced block in spermatogonial differentiation in F1 hybrids of Lewis and Brown-Norway rats. Although addition of exogenous testosterone restores the block, it also raises FSH, and hence it had not been possible to conclusively determine which hormone was inhibiting spermatogonial differentiation. In the present study, we establish the relative roles of testosterone and FSH in this inhibition using three different approaches. The first approach involved the treatment of irradiated rats, in which differentiation was stimulated by GnRH-ant plus flutamide, with FSH for 2 wk; the FSH reduced the percentage of tubules that were differentiated (TDI) by about 2 fold, indicating that FSH does have an inhibitory role. The second approach involved treatment of irradiated, hypophysectomized rats with exogenous testosterone for 10 wk; testosterone also reduced the TDI, demonstrating that testosterone had a definite inhibitory effect, independent of pituitary hormones. Furthermore, in this protocol we showed that TDI in the hypophysectomized testosterone-treated group, which had higher intratesticular testosterone levels but lacked FSH, was slightly higher than the TDI in a GnRH-antagonist testosterone-treated group of irradiated rats, which had normal physiological levels of FSH; this result supports a role for endogenous FSH in suppressing spermatogonial differentiation in the latter group. The third approach involved injection of an active anti-FSH antibody for 10 d in untreated, GnRH-ant plus flutamide-treated, or GnRH-ant plus testosterone-treated irradiated rats. This was not sufficient to increase the TDI. However, flutamide given in a similar treatment schedule did increase the TDI in GnRH-ant plus testosterone-treated rats. We conclude that both testosterone and FSH individually inhibit spermatogonial differentiation after irradiation, but testosterone is a more highly potent inhibitor than is FSH. PMID- 16210367 TI - Differential effects of central leptin, insulin, or glucose administration during fasting on the hypothalamic-pituitary-thyroid axis and feeding-related neurons in the arcuate nucleus. AB - The reductions in circulating levels of leptin, insulin, and glucose with fasting serve as important homeostasis signals to neurons of the hypothalamic arcuate nucleus that synthesize neuropeptide Y (NPY)/agouti-related protein (AGRP) and alpha-MSH/cocaine and amphetamine-regulated transcript. Because the central administration of leptin is capable of preventing the inhibitory effects of fasting on TRH mRNA in hypophysiotropic neurons primarily through effects on the arcuate nucleus, we determined whether the continuous administration of 30 mU/d insulin or 648 microg/d glucose into the cerebrospinal fluid by osmotic minipump might also have similar effects on the hypothalamic-pituitary-thyroid axis. As anticipated, the intracerebroventricular infusion of leptin reduced fasting induced elevations in NPY and AGRP mRNA and increased proopiomelanocortin and cocaine and amphetamine-regulated transcript mRNA in the arcuate nucleus. In addition, leptin prevented fasting-induced reduction in pro-TRH mRNA levels in the paraventricular nucleus and in circulating thyroid hormone levels. In contrast, whereas insulin increased proopiomelanocortin mRNA and both insulin and glucose reduced NPY mRNA in arcuate nucleus neurons, neither prevented the fasting-induced suppression in hypophysiotropic TRH mRNA or circulating thyroid hormone levels. We conclude that insulin and glucose only partially replicate the central effects of leptin and may not be essential components of the hypothalamic pituitary-thyroid regulatory system during fasting. PMID- 16210368 TI - Retinoic acid metabolism and signaling pathways in the adult and developing mouse testis. AB - As a first step in investigating the role of retinoic acid (RA) in mouse testis, we analyzed the distribution pattern of the enzymes involved in vitamin A storage (lecithin:retinol acyltransferase), RA synthesis (beta-carotene 15,15' monoxygenase and retinaldehyde dehydrogenases) and RA degradation (cytochrome P450 hydroxylases) as well as those of all isotypes of receptors transducing the RA signal [RA receptors (RARs) and rexinoid receptors (RXRs)]. Our data indicate that in adult testis 1) cytochrome P450 hydroxylase enzymes may generate in peritubular myoid cells a catabolic barrier that prevents circulating RA and RA synthesized by Leydig cells to enter the seminiferous epithelium; 2) the compartmentalization of RA synthesis within this epithelium may modulate, through paracrine mechanisms, the coupling between spermatogonia proliferation and spermatogenesis; 3) retinyl esters synthesized in round spermatids by lecithin:retinol acyltransferase may be transferred and stored in Sertoli cells, in the form of adipose differentiation-related protein-coated lipid droplets. We also show that RARalpha and RXRbeta are confined to Sertoli cells, whereas RARgamma is expressed in spermatogonia and RARbeta, RXRalpha, and RXRgamma are colocalized in step 7-8 spermatids. Correlating these expression patterns with the pathological phenotypes generated in response to RAR and RXR mutations and to postnatal vitamin A deficiency suggests that spermiation requires RXRbeta/RARalpha heterodimers in Sertoli cells, whereas spermatogonia proliferation involves, independently of RXR, two distinct RAR-mediated signaling pathways in both Sertoli cells and spermatogonia. Our data also suggest that the involvement of RA in testis development starts when primary spermatogonia first appear. PMID- 16210369 TI - Differential effects of estradiol, progesterone, and testosterone on vaginal structural integrity. AB - Ovarian steroids are known to be important in maintaining vaginal tissue, and evidence is mounting that imbalances in the hormonal milieu contribute to vaginal pathophysiology. To date, limited data are available on the effects of hormone deprivation and replacement on vaginal tissue morphology and vaginal innervation. The goal of this study was to assess the dynamic changes in vaginal tissue structure in response to sex steroid hormone deprivation and administration. Female Sprague-Dawley rats were either kept intact (controls) or ovariectomized. Ovariectomized animals were treated with vehicle, estradiol, testosterone, progesterone, or a combination of estradiol plus testosterone or progesterone. Histological techniques, including stereological analysis and immunohistochemistry for localization of neuronal markers, were used. Ovariectomy produced a significant decrease in epithelial height that was restored with estradiol replacement. Interestingly, a subphysiological dose of estradiol resulted in hyperplasia of the vaginal epithelium and nonvascular smooth muscle. Neither testosterone nor progesterone had a significant effect on epithelial height or muscularis thickness. However, testosterone treatment resulted in a significant increase in small adrenergic nerve fibers. Addition of either testosterone or progesterone to estradiol mitigated but did not abolish the effects of estradiol alone. This study demonstrates that estradiol and testosterone have differential effects on vaginal tissue parameters and that ovarian hormones are critical for the maintenance of genital tissue structure. Present observations also suggest that combined replacement regimens may be required for an optimal physiological response. PMID- 16210370 TI - The Leu34Phe ProCART mutation leads to cocaine- and amphetamine-regulated transcript (CART) deficiency: a possible cause for obesity in humans. AB - Cocaine- and amphetamine-regulated transcript (CART) is an anorexigenic neuropeptide synthesized in the hypothalamus. A Leu34Phe missense mutation in proCART has been found in an obese family in humans. Here we show that humans bearing the Leu34Phe mutation in proCART have severely diminished levels of bioactive CART, but elevated amounts of partially processed proCART in their serum. Expression of wild-type proCART in AtT-20 cells showed that it was sorted to the regulated secretory pathway, a necessity for proper processing to bioactive CART. However, expressed Leu34Phe proCART was missorted, poorly processed, and secreted constitutively. The defective intracellular sorting of Leu34Phe proCART would account for the reduced levels of bioactive CART in affected humans. These results suggest that the obesity observed in humans bearing the Leu34Phe mutation could be due to a putative deficiency in hypothalamic bioactive CART. PMID- 16210371 TI - Prevention of programmed hyperleptinemia and hypertension by postnatal dietary omega-3 fatty acids. AB - Fetal programming is now recognized as a key determinant of the adult phenotype, with major implications for adult-onset diseases including hypertension. Two mediators of fetal programming are maternal nutrition and fetal glucocorticoid exposure. Recent studies show that postnatal dietary manipulations can exacerbate programming effects, but whether programming effects can be attenuated by postnatal dietary manipulations, and thus provide a possible therapeutic strategy, is unknown. In this study, we tested the hypothesis that a postnatal diet enriched with long-chain omega-3 fatty acids attenuates programmed hyperleptinemia and hypertension. Pregnant rats were treated with dexamethasone (Dex) from d 13 to term, and offspring were cross-fostered to mothers on either a standard diet or a diet high in omega-3 fatty acids and remained on these diets postweaning. Maternal Dex reduced birthweight and delayed the onset of puberty in offspring. Hyperleptinemia (associated with elevated leptin mRNA expression in adipose tissue) and hypertension were evident in offspring by 6 months of age in Dex-exposed animals consuming a standard diet, but these effects were completely blocked by a high omega-3 diet. These results demonstrate for the first time that manipulation of postnatal diet can limit adverse outcomes of fetal programming, with programmed hyperleptinemia and hypertension prevented by a postnatal diet enriched with omega-3 fatty acids. This raises the possibility that dietary supplementation with omega-3 fatty acids may provide a viable therapeutic option for preventing and/or reducing adverse programming outcomes in humans. PMID- 16210372 TI - Changes in hypothalamic corticotropin-releasing hormone, neuropeptide Y, and proopiomelanocortin gene expression during chronic rapid eye movement sleep deprivation of rats. AB - Chronic rapid eye movement (paradoxical) sleep deprivation (REM-SD) of rats leads to two conspicuous pathologies: hyperphagia coincident with body weight loss, prompted by elevated metabolism. Our goals were to test the hypotheses that 1) as a stressor, REM-SD would increase CRH gene expression in the hypothalamus and that 2) to account for hyperphagia, hypothalamic gene expression of the orexigen neuropeptide Y (NPY) would increase, but expression of the anorexigen proopiomelanocortin (POMC) would decrease. Enforcement of REM-SD of adult male rats for 20 d with the platform (flowerpot) method led to progressive hyperphagia, increasing to approximately 300% of baseline; body weight steadily declined by approximately 25%. Consistent with changes in food intake patterns, NPY expression rapidly increased in the hypothalamic arcuate nucleus by d 5 of REM-SD, peaking at d 20; by contrast, POMC expression decreased progressively during REM-SD. CRH expression was increased by d 5, both in mRNA and ability to detect neuronal perikaryal staining in paraventricular nucleus with immunocytochemistry, and it remained elevated thereafter with modest declines. Taken together, these data indicate that changes in hypothalamic neuropeptides regulating food intake are altered in a manner consistent with the hyperphagia seen with REM-SD. Changes in CRH, although indicative of REM-SD as a stressor, suggest that the anorexigenic actions of CRH are ineffective (or disabled). Furthermore, changes in NPY and POMC agree with current models of food intake behavior, but they are opposite to their acute effects on peripheral energy metabolism and thermogenesis. PMID- 16210373 TI - Female sex steroids increase adrenomedullin-induced vasodilation by increasing the expression of adrenomedullin2 receptor components in rat mesenteric artery. AB - Based on the favorable effects of female sex steroids in vascular functions and the potent hypotensive effects of adrenomedullin (AM), we hypothesized that AM induced vasodilation is gender dependent, and female sex steroids enhance this effect. In endothelium-intact rat mesenteric artery, AM (1 nm-0.3 microM)-induced concentration-dependent relaxation was significantly (P < 0.05) higher in females [pD2(-log EC50 of the molar concentration), 7.05 +/- 0.10; maximal relaxation response (Emax), 69.2 +/- 3.46%] than males (pD2, 6.53 +/- 0.08; Emax, 53.28 +/- 4.86%). The increased relaxation was lost when the females were ovariectomized (OVX) (pD2, 6.14 +/- 0.24; Emax, 39.68 +/- 5.68%). The reduced relaxation response in OVX rats was reversed by administration of either progesterone (P4; pD2, 7.18 +/- 0.07; Emax, 72.4 +/- 2.76%) or 17beta-estradiol (E2; pD2, 7.00 +/- 0.14; Emax, 70.4 +/- 4.79%). AM mediates its effects through either AM(22-52) sensitive AM1 receptors [composed of calcitonin receptor-like receptors (CLs) and receptor activity-modifying protein (RAMP)2] or AM2 receptors (CL/RAMP3), which can be antagonized more potently by calcitonin gene-related peptide(8-37) than AM(22-52). Pharmacological characterization suggested the involvement of AM2 receptors in the increased vasodilatory effect of AM in both P4- and E2-treated animals as calcitonin gene-related peptide(8-37) (10 microM) was more potent in antagonizing the AM effects (Emax, P(4): 25.92 +/- 5.32%; E2: 29.11 +/- 7.41%) than AM(22-52) (100 microM). RT-PCR studies also supported the involvement of AM2 receptors because expression of mRNA levels encoding CL (previously reported) and RAMP3 were increased in P4- or E2-treated OVX rats. In conclusion, AM-induced vasodilation is gender-dependent and increased by female sex steroids by increased expression of AM2 receptor components. PMID- 16210374 TI - Obesity-related glomerulopathy: insights from gene expression profiles of the glomeruli derived from renal biopsy samples. AB - Obesity-related glomerulopathy (ORG) is an important complication of obesity. The pathophysiological mechanism of glomerular injury in ORG is incompletely understood. Gene expression profiles in the glomeruli obtained from renal biopsy samples of patients with ORG were investigated, using a microdissection technique combined with Affymetrix microarray analysis. Six patients presented with obesity, proteinuria, and biopsy-proven ORG were enrolled. Two sex- and age matched donor kidneys were applied as the controls. Glomeruli were dissected out from renal biopsy samples under microscope, and total RNA was extracted using RNeasy Micro kit. After two rounds of T7 promoter-based RNA amplification, gene expression profiles of the glomeruli samples were detected using Affymetrix U133A gene chips. Bioinformatic tools were applied to analyze the microarray data. Results of candidate ORG-related genes were further confirmed by real-time quantitative PCR and immunohistochemistry staining using renal biopsy samples of a larger pool of 15 ORG patients. Genes related to lipid metabolism, inflammatory cytokines, and insulin resistance were the most highlighted subgroups that significantly changed in the glomerular gene expression profiles of the ORG patients, compared with the controls. The expression levels of several key genes in lipid metabolism were increased over 2-fold, including low-density lipoprotein receptor, fatty acid binding protein 3, and sterol regulatory element binding protein 1. Moreover, some inflammatory cytokines and their downstream molecules were increased as well, including TNF-alpha and its receptors, IL-6 signal transducer, and interferon-gamma. As the indicators of insulin resistance in the local glomerular cells, levels of glucose-transporter 1, leptin receptor, peroxisome proliferator-activated receptor-gamma, and vascular endothelial growth factor increased, too. In addition to lipid dysmetabolism and insulin resistance, the activation of an inflammatory process in the glomeruli might play a unique role in the development of obesity-related glomerulopathy. Our results expand the understanding of obesity-induced glomerular injuries and shed light on new approaches in the treatment of this disease. PMID- 16210375 TI - Endocrine gland-derived vascular endothelial growth factor is expressed in human peri-implantation endometrium, but not in endometrial carcinoma. AB - Endocrine gland-derived vascular endothelial growth factor (EG-VEGF) is a newly identified angiogenic and permeability-enhancing factor, predominantly expressed in steroidogenic tissues. Recently, we found that EG-VEGF is also expressed in the normal peri-implantation endometrial samples from patients of reproductive ages (80%). Immunohistochemistry analysis showed that EG-VEGF is predominantly expressed in the glandular epithelial cells and its expression is dynamic during the menstrual cycle with a peak expression at the mid-luteal phase. We also found that EG-VEGF transcripts are up-regulated in all the peri-implantation endometrial samples from the patients after the ovulating dose of human chorionic gonadotropin in gonadotropin-stimulated cycles and patients receiving hormone replacement therapy. In in vitro endometrial cell culture, EG-VEGF mRNA was detected in endometrial cells only in the presence of steroids, suggesting that EG-VEGF expression is highly dependent on the steroid hormones. Subsequent expression analyses on the EG-VEGF receptors showed that hPK-R1 and hPK-R2 are differentially expressed in human endometrium, but show no significant correlation with the hormonal treatments. On the other hand, EG-VEGF transcript was rarely detected in the endometrial samples from the postmenopausal patients and patients with endometrial carcinoma. It may imply that EG-VEGF may only play a role in vascular function of peri-implantation endometrium, but is unlikely to be associated with the etiology of endometrial cancer development. PMID- 16210376 TI - Tyrosine kinase and mitogen-activated protein kinase/extracellularly regulated kinase differentially regulate intracellular calcium concentration responses to angiotensin II/III and bradykinin in rat cortical thick ascending limb. AB - The cortical thick ascending limb (CTAL) coexpresses angiotensin (Ang) II/Ang III receptor type 1A (AT(1A)-R) and bradykinin (BK) receptor type 2 (B2-R). In several cell types, these two receptors share the same signaling pathways, although their physiological functions are often opposite. In CTAL, little is known about the intracellular transduction events leading to the final physiological response induced by these two peptides. We investigated and compared in this segment the action of Ang II/III and BK on intracellular calcium concentration ([Ca2+]i) response and metabolic CO2 production, an index of Na+ transport, by using inhibitors of protein kinase C (bisindolylmaleimide), Src tyrosine kinase (herbimycin A and PP2), and MAPK/ERK (PD98059 and UO126). Ang II/III and BK (10(-7) mol/liter) released Ca2+ from the same intracellular pools but activated different Ca2+ entry pathways. Ang II/III- or BK-induced [Ca2+]i increases were similarly potentiated by bisindolylmaleimide. Herbimycin A and PP2 decreased similarly the [Ca2+]i responses induced by Ang II/III and BK. In contrast, PD98059 and UO126 affected the effects of BK to a larger extent than those of Ang II/III. Especially, the Ca2+ influx induced by BK was more strongly inhibited than that induced by Ang II/III in the presence of both compounds. The Na+ transport was inhibited by BK and stimulated by Ang II/III. The inhibitory action of BK on Na+ transport was blocked by UO126, whereas the stimulatory response of Ang II/III was potentiated by UO126 but blocked by bisindolylmaleimide. These data suggest that the inhibitory effect of BK on Na+ transport seems to be directly mediated by an increase in Ca2+ influx dependent on MAPK/ERK pathway activation. In contrast, the stimulatory effect of Ang II/III on Na+ transport is more complex and involves PKC and MAPK/ERK pathways. PMID- 16210378 TI - Can urban regeneration programmes assist coping and recovery for people with mental illness? Suggestions from a qualitative case study. AB - Researchers and policy-makers are increasingly recognizing that urban socio environmental conditions can affect the development and course of numerous health problems. The aim of this paper is to investigate the impact an urban regeneration programme can have on everyday functioning, coping and recovery for people with a mental illness. We were also interested in discerning which component parts of the regeneration are the most important in positively affecting people with mental illness. These questions were explored through an in depth qualitative case study of the Gospel Oak neighbourhood in London, which recently underwent an intensive urban regeneration programme. Interviews and focus groups were conducted with residents living with a mental illness (n = 16). Relevant participant observation was also conducted. Participants reported that interventions that improved community safety were by far the most important in affecting everyday coping and functioning. Interventions that improved the quantity and quality of shared community facilities had a positive, but milder effect on mental health. Component parts that appeared to have little effect included environmental landscaping and greater community involvement in decision making processes. Most participants reported that their mental illness was a consequence of severe insults over the life-span, for example childhood neglect or family breakdown. Thus, the regeneration was seen as something that could assist coping, but not something that could significantly contribute to complete recovery. Our results thus suggest that urban regeneration can have a mild impact on people with mental illness, but this appears to be outweighed by life-span experience of severe individual-level risk factors. That said, some of our findings converge with other studies indicating that community safety and community facilities can play a role in positively affecting mental health. Further ethnographic and epidemiological research is necessary to explore these two factors. PMID- 16210377 TI - Testosterone inhibits adipogenic differentiation in 3T3-L1 cells: nuclear translocation of androgen receptor complex with beta-catenin and T-cell factor 4 may bypass canonical Wnt signaling to down-regulate adipogenic transcription factors. AB - Testosterone supplementation in men decreases fat mass; however, the mechanisms by which it inhibits fat mass are unknown. We hypothesized that testosterone inhibits adipogenic differentiation of preadipocytes by activation of androgen receptor (AR)/beta-catenin interaction and subsequent translocation of this complex to the nucleus thereby bypassing canonical Wnt signaling. We tested this hypothesis in 3T3-L1 cells that differentiate to form fat cells in adipogenic medium. We found that these cells express AR and that testosterone and dihydrotestosterone dose-dependently inhibited adipogenic differentiation as analyzed by Oil Red O staining and down-regulation of CCAAT/enhancer binding protein-alpha and -delta and peroxisome proliferator-activated receptor-gamma2 protein and mRNA. These inhibitory effects of androgens were partially blocked by flutamide or bicalutamide. Androgen treatment was associated with nuclear translocation of beta-catenin and AR. Immunoprecipitation studies demonstrated association of beta-catenin with AR and T-cell factor 4 (TCF4) in the presence of androgens. Transfection of TCF4 cDNA inhibited adipogenic differentiation, whereas a dominant negative TCF4 cDNA construct induced adipogenesis and blocked testosterone's inhibitory effects. Our gene array analysis indicates that testosterone treatment led to activation of some Wnt target genes. Expression of constitutively activated AR fused with VP-16 did not inhibit the expression of CCAAT/enhancer binding protein-alpha in the absence of androgens. Testosterone and dihydrotestosterone inhibit adipocyte differentiation in vitro through an AR mediated nuclear translocation of beta-catenin and activation of downstream Wnt signaling. These data provide evidence for a regulatory role for androgens in inhibiting adipogenic differentiation and a mechanistic explanation consistent with the observed reduction in fat mass in men treated with androgens. PMID- 16210379 TI - Two single-nucleotide polymorphisms in the human vitamin D receptor promoter change protein-DNA complex formation and are associated with height and vitamin D status in adolescent girls. AB - Numerous association studies have dealt with single-nucleotide polymorphisms (SNPs) in coding and intronic regions of the human vitamin D receptor (hVDR) gene. We have hypothesized that phenotypic traits may also be associated with variations in VDR expression due to the presence of SNPs in promoter regions. In this work, we have studied two SNPs located 1521 bp (G/C) and 1012 bp (A/G) upstream of the transcriptional start site of the main human VDR gene promoter. One base-change in any of the two variant sites led to a dramatic change in protein-DNA complex formation using nuclear extracts from HEK293, Caco-2 and COS 7 cells. Genetic analysis of 185 healthy adolescent girls evidenced two major haplotypes: 1521G/1012A and 1521C/1012G and three main genotypes: homozygous for 1521G/1012A (21.1%), homozygous for 1521C/1012G (17.3%) and heterozygous 1521CG/1012GA (57.3%). On the basis of transfection data, promoter activity was nearly 2-fold higher with the 1521G/1012A haplotype, when compared with the 1521C/1012G haplotype. Clinical and biological association study in the adolescent cohort showed that girls with a CC/GG genotype had (i) lower circulating levels of 25-dihydroxyvitamin D, with no detectable consequence on calcium metabolism, (ii) lower serum IGF-1 levels and (iii) smaller height from 11 years of age up to adult height. PMID- 16210380 TI - Mutant SPTLC1 dominantly inhibits serine palmitoyltransferase activity in vivo and confers an age-dependent neuropathy. AB - Mutations in enzymes involved in sphingolipid metabolism and trafficking cause a variety of neurological disorders, but details of the molecular pathophysiology remain obscure. SPTLC1 encodes one subunit of serine palmitoyltransferase (SPT), the rate-limiting enzyme in sphingolipid synthesis. Mutations in SPTLC1 cause hereditary sensory and autonomic neuropathy (type I) (HSAN1), an adult onset, autosomal dominant neuropathy. HSAN1 patients have reduced SPT activity. Expression of mutant SPTLC1 in yeast and mammalian cell cultures dominantly inhibits SPT activity. We created transgenic mouse lines that ubiquitously overexpress either wild-type (SPTLC1(WT)) or mutant SPTLC1 (SPTLC1(C133W)). We report here that SPTLC1(C133W) mice develop age-dependent weight loss and mild sensory and motor impairments. Aged SPTLC1(C133W) mice lose large myelinated axons in the ventral root of the spinal cord and demonstrate myelin thinning. There is also a loss of large myelinated axons in the dorsal roots, although the unmyelinated fibers are preserved. In the dorsal root ganglia, IB4 staining is diminished, whereas expression of the injury-induced transcription factor ATF3 is increased. These mice represent a novel mouse model of peripheral neuropathy and confirm the link between mutant SPT and neuronal dysfunction. PMID- 16210381 TI - Spatial expression of germ cell markers during maturation of human fetal male gonads: an immunohistochemical study. AB - BACKGROUND: The aim of the present study was to examine fetal male germ cells for expression of proteins associated with differentiation and maturation and to compare them with morphologically defined subpopulations. METHODS: Testes of 61 fetuses from week 12 of gestation to the newborn period were selected. Immunohistochemistry was performed using antibodies to proteins associated with differentiation of germ cells (c-KIT, AP-2gamma) or pluripotency (OCT3/4), oncofetal protein M2A and spermatogonial marker MAGE-A4. RESULTS: Two subtypes of fetal germ cells were detected by quantification and immunohistochemistry. Nearly all germ cells with morphological criteria of gonocytes and intermediate cells co expressed OCT3/4, c-KIT, M2A and AP-2gamma. Starting from week 12, their number increased up to week 18/19 and then declined continuously during further development. After week 25, pre-spermatogonia were predominant and expressed MAGE A4 selectively. CONCLUSIONS: Fetal male germ cells are comprised of two major groups with distinct immunohistochemical phenotypes. Germ cells that are predominantly found before week 25 of gestation co-express oncofetal proteins OCT3/4, c-KIT, M2A and AP-2gamma. After week 25, most germ cells have lost their pluripotent potential and acquire a spermatogonial phenotype defined by expression of MAGE-A4. PMID- 16210382 TI - Is there an association between septate uterus and endometriosis? AB - BACKGROUND: The question of whether Mullerian anomalies are significantly more often combined with endometriosis is a controversially discussed problem. Some publications described this association in patients with obstructive but not non obstructive Mullerian anomalies or controls without Mullerian anomalies. The aim of this study was to evaluate the incidence of endometriosis in patients with a septate uterus as a non-obstructive form of Mullerian anomalies. METHODS: In a retrospective study, we evaluated 120 patients (29.4 +/- 4.7 years; mean +/- SD) with a septate uterus. The control group consisted of 486 consecutive infertile patients (30.8 +/- 6.3 years) with a normal hysteroscopy and laparoscopy. RESULTS: The incidence of dysmenorrhoea was comparable in both groups, but the incidence of endometriosis was significantly higher in patients with a septate uterus (25.8 versus 15.2%, P = 0.006). CONCLUSION: Our results suggest a higher incidence of endometriosis in patients with a septate uterus. If it can be confirmed by others, the initial finding of a septate uterus in infertile patients should be followed by a combined hysteroscopy and laparoscopy. PMID- 16210383 TI - Cryopreservation of intact testicular tissue from boys with cryptorchidism. AB - BACKGROUND: Boys with cryptorchidism often face fertility problems in adult life despite having orchiopexy performed at a very young age. During this operation, a biopsy of the testis is normally taken in order to evaluate their infertility potential and the presence of malignant cells. This study evaluated the morphology and functional capacity of cryopreserved testes biopsies and their possible use in fertility preservation. METHODS: Biopsies from 11 testes (eight boys) were obtained. Each biopsy was subdivided into six pieces and two pieces were frozen in each of two different cryoprotectants. One fresh and two cryopreserved pieces were cultured for 2 weeks. All pieces were prepared for histology. Used culture media were analysed for testosterone and inhibin B concentrations. RESULTS: The morphology of the fresh and frozen-thawed samples was similar, with well-preserved seminiferous tubules and interstitial cells. A similar picture appeared after 2 weeks of culture, but a few of the cultured biopsies contained small necrotic areas. The presence of spermatogonia was verified by c-kit-positive immunostaining. Production of testosterone and inhibin B (ng/mm(3) testis tissue) in the frozen-thawed pieces was on average similar to that of the fresh samples. CONCLUSIONS: Intact testicular tissue from young boys with non-descended testes tolerates cryopreservation with surviving spermatogonia and without significant loss of the ability to produce testis-specific hormones in vitro. It may be an option to freeze part of the testis biopsy, which is routinely removed during the operation for cryptorchidism, for fertility preservation in adult life. PMID- 16210384 TI - Combined estrogen receptor alpha and estrogen receptor beta genotypes influence the age of menarche. AB - BACKGROUND: Age at menarche has a strong genetic influence. We reported recently an association between the XbaI (351A-->C)and PvuII (397T-->C) polymorphisms of the estrogen receptor (ER)alpha gene with the age of menarche in Greek adolescents. In the present study, we examined whether ERbeta genotypes alone, or in combination with ERalpha genotypes, may also influence onset of menarche. METHODS: We performed genotyping for the single nucleotide polymorphisms 1730A- >G and 1082G-->A of the ERbeta gene and examined their association with the age of menarche in the same cohort of 145 Greek girls. We then looked for a possible effect of combined ERalpha and beta genotypes on the age of menarche. RESULTS: Menarche occurred 7 months later in girls with the AA genotype of the 1730A-->G polymorphism than in girls with the AG genotype (mean +/- SD: 13.23 +/- 1.24 versus 12.66 +/- 1.26 years, respectively; P = 0.005). The 1082G-->A polymorphism was not detected in any of the girls examined. A significant effect of combined ERalpha and beta genotypes was also apparent. Menarche occurred 11 months later in girls bearing the AA/TT,AA (ERalpha, ERbeta) genotypes compared with girls with the CC/CC,AG genotype (13.30 +/- 1.27 nersus 12.41 +/- 1.28 years; P = 0.042). The difference remained significant after adjusting for body mass index (P = 0.034). CONCLUSION: Combined ERalpha and ERbeta polymorphisms may influence the age of menarche. PMID- 16210385 TI - The ATAC ('Arimidex', Tamoxifen, Alone or in Combination) adjuvant breast cancer trial: first results of the endometrial sub-protocol following 2 years of treatment. AB - BACKGROUND: Tamoxifen treatment results in a doubling of the risk of endometrial cancer after 1-2 years of treatment and a quadrupling after 5 years. Anastrozole, a third-generation aromatase inhibitor, with superior efficacy to tamoxifen, may also offer tolerability benefits in terms of effects on the endometrium. METHODS AND RESULTS: A sub-protocol of the ATAC trial compared the incidence/type of intrauterine changes following treatment with these agents in a subgroup of patients (n = 285) from the main trial. After 2 years anastrozole treatment, endometrial thickness remained 0.10). Patients with T2DM exhibited an increased internal temperature threshold for the onset of vasodilation at both untreated and BT-treated sites. At BT treated sites, T or thresholds were 36.28 +/- 0.07 degrees C in controls and 36.55 +/- 0.05 degrees C in T2DM patients (P < 0.05), indicating delayed onset of active vasodilation in patients. Sensitivity of vasodilation was variable in both groups, with no consistent difference between groups (P > 0.05). We conclude that altered control of active cutaneous vasodilation may contribute to impaired thermoregulation in patients with T2DM. PMID- 16210433 TI - Effects of polymorphisms in beta1-adrenoceptor and alpha-subunit of G protein on heart rate and blood pressure during exercise test. The Finnish Cardiovascular Study. AB - We tested whether the Arg389Gly and Ser49Gly polymorphisms of the beta1 adrenergic receptor gene ADRB1 and the T393C polymorphism of the G protein alpha subunit gene GNAS1 modulate heart rate (HR) and blood pressure responses during an exercise stress test. The study population comprised 890 participants (563 men and 327 women, mean age 58.1 +/- 12.6 yr) of the Finnish Cardiovascular Study. Their HR, systolic (SAP), and diastolic arterial pressures (DAP) at rest, during exercise, and 4 min after the test were measured and analyzed by repeated measurement ANOVA (RANOVA). Genotypes were detected by TaqMan 5' nuclease assay. In all subjects, and in men and women separately, the T393C of GNAS1 was the only polymorphism with genotype x time interaction in HR over the three study phases (P = 0.04, RANOVA). None of the polymorphisms presented genotype x time interaction in SAP or DAP responses (P > 0.10, RANOVA). In all subjects at rest, the Ser49Gly polymorphism of ADRB1 tended (P = 0.06, ANOVA) to differentiate HR. Arg389Gly polymorphism of ADRB1 affected maximal SAP during exercise (P = 0.04, ANOVA) and the change in SAP from rest to maximal (P = 0.03, ANOVA). Arg389 homozygotes, particularly men, were less likely to have ventricular extrasystoles during the exercise (odds ratio = 0.68, 95% confidence interval = 0.51-0.91, P = 0.009, and odds ratio = 0.60, 95% confidence interval = 0.42-0.86, P = 0.006, respectively) than did Gly389 carriers. In conclusion, polymorphisms examined appear to have modulatory effects on hemodynamics in a clinical exercise test setting. However, the effects in absolute numbers were minor and clinically possibly insignificant. PMID- 16210434 TI - Effects of obesity and sex on the energetic cost and preferred speed of walking. AB - The metabolic energy cost of walking is determined, to a large degree, by body mass, but it is not clear how body composition and mass distribution influence this cost. We tested the hypothesis that walking would be most expensive for obese women compared with obese men and normal-weight women and men. Furthermore, we hypothesized that for all groups, preferred walking speed would correspond to the speed that minimized the gross energy cost per distance. We measured body composition, maximal oxygen consumption, and preferred walking speed of 39 (19 class II obese, 20 normal weight) women and men. We also measured oxygen consumption and carbon dioxide production while the subjects walked on a level treadmill at six speeds (0.50-1.75 m/s). Both obesity and sex affected the net metabolic rate (W/kg) of walking. Net metabolic rates of obese subjects were only approximately 10% greater (per kg) than for normal-weight subjects, and net metabolic rates for women were approximately 10% greater than for men. The increase in net metabolic rate at faster walking speeds was greatest in obese women compared with the other groups. Preferred walking speed was not different across groups (1.42 m/s) and was near the speed that minimized gross energy cost per distance. Surprisingly, mass distribution (thigh mass/body mass) was not related to net metabolic rate, but body composition (% fat) was (r2= 0.43). Detailed biomechanical studies of walking are needed to investigate whether obese individuals adopt novel energy saving mechanisms during walking. PMID- 16210435 TI - Vanilloid type 1 receptor and the acid-sensing ion channel mediate acid phosphate activation of muscle afferent nerves in rats. AB - Reflex cardiovascular responses to contracting skeletal muscle are mediated by mechanical and metabolic stimulation of thin-fiber muscle afferents. Diprotonated phosphate (H2PO4-) excites those thin-fiber nerves and evokes the muscle pressor reflex. The receptors mediating this response are unknown. Thus we examined the role played by purinergic receptors, vanilloid type 1 receptors (VR1), and acid sensing ion channels (ASIC) in mediating H2PO4- -evoked pressor responses. Phosphate and blocking agents were injected into the arterial blood supply of the hindlimb muscles of 53 decerebrated rats. H2PO4- (86 mM, pH 6.0) increased mean arterial pressure by 25 +/- 2 mmHg, whereas monoprotonated phosphate (HPO4(2-), pH 7.5) had no effect. Pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (a purinergic receptor antagonist, 2 mM) did not block the response. However, capsazepine (a VR1 antagonist, 1 mg/kg) attenuated the reflex by 60% and amiloride (an ASIC blocker, 6 microg/kg) by 52%. Of note, the H2PO4- -induced pressor response was attenuated by 87% when both capsazepine and amiloride were injected before the H2PO4-. In conclusion, VR1 and ASIC mediate the pressor response due to H2PO4-. The H2PO4- -evoked response was greater when VR1 and ASIC blockers were given simultaneously than when the respective blockers were given separately. Our laboratory's previous study has shown that H+ stimulates ASIC (but not VR1) on thin-fiber afferent nerves in evoking the reflex response. Thus VR1 and ASIC are likely to play a coordinated and interactive role in processing the muscle afferent response to H2PO4-. Furthermore, the physiological mechanisms mediating the response to H+ and H2PO4- are likely to be different. PMID- 16210436 TI - The influence of radial RBC distribution, blood velocity profiles, and glycocalyx on coupled NO/O2 transport. AB - The purpose of this investigation was to study the effect of the presence of red blood cells (RBCs) in the plasma layer near the arteriole wall on nitric oxide (NO) and oxygen (O2) transport. To this end, we extended a coupled NO and O2 diffusion-reaction model in the arteriole, developed by our group, to include the effect of the presence of RBCs in the plasma layer and the effect of convection. Two blood flow velocity profiles (plug and parabolic) were tested. The average hematocrit in the bloodstream was assumed to be constant in the central core and decreasing to zero in the boundary layer next to the endothelial surface layer. The effect of the presence or absence of RBCs near the endothelium was studied while varying the endothelial surface layer and boundary layer thickness. With RBCs present in the boundary layer, the model predicts that 1) NO decreases significantly in the endothelium and vascular wall; 2) there is a very small increase in endothelial and vascular wall Po2; 3) scavenging of NO by hemoglobin decreases with increasing thickness of the boundary layer; 4) the shape of the velocity profile influences both NO and Po2 gradients in the bloodstream; and 5) the presence of RBCs in the boundary layer near the endothelium has a much larger effect on NO than on O2 transport. PMID- 16210437 TI - Regulation of synaptic vesicles pools within motor nerve terminals during short term facilitation and neuromodulation. AB - The reserve pool (RP) and readily releasable pool (RRP) of synaptic vesicles within presynaptic nerve terminals were physiologically differentiated into distinctly separate functional groups. This was accomplished in glutamatergic nerve terminals by blocking the glutamate transporter with dl-threo-beta benzyloxyaspartate (TBOA; 10 microM) during electrical stimulation with either 40 Hz of 10 pulses within a train or 20- or 50-Hz continuous stimulation. The 50-Hz continuous stimulation decreased the excitatory postsynaptic potential amplitude 60 min faster than for the 20-Hz continuous stimulation in the presence of TBOA (P < 0.05). There was no significant difference between the train stimulation and 20-Hz continuous stimulation in the run-down time in the presence of TBOA. After TBOA-induced synaptic depression, the excitatory postsynaptic potentials were rapidly (<1 min) revitalized by exposure to serotonin (5-HT, 1 microM) in every preparation tested (P < 0.05). At this glutamatergic nerve terminal, 5-HT promotes an increase probability of vesicular docking and fusion. Quantal recordings made directly at nerve terminals revealed smaller quantal sizes with TBOA exposure with a marked increase in quantal size as well as a continual appearance of smaller quanta upon 5-HT treatment after TBOA-induced depression. Thus 5-HT was able to recruit vesicles from the RP that were not rapidly depleted by acute TBOA treatment and electrical stimulation. The results support the notion that the RRP is selectively activated during rapid electrical stimulation sparing the RP; however, the RP can be recruited by the neuromodulator 5-HT. This suggests at least two separate kinetic and distinct regulatory paths for vesicle recycling within the presynaptic nerve terminal. PMID- 16210438 TI - Surfactant replacement partially restores the activity of pulmonary stretch receptors in surfactant-depleted cats. AB - Single units of slowly adapting pulmonary stretch receptors (PSRs) were investigated in anesthetized cats during spontaneous breathing on continuous positive airway pressure (2-5 cmH2O), before and after lung lavage and then after instillation of surfactant to determine the PSR response to surfactant replacement. PSRs were classified as high threshold (HT) and low threshold (LT), and their instantaneous impulse frequency (f imp) was related to transpulmonary pressure (Ptp) and tidal volume (Vt). Both the total number of impulses and maximal f imp of HT and LT PSRs decreased after lung lavage (55 and 45%, respectively) in the presence of increased Ptp and decreased Vt. While Ptp decreased markedly and Vt remained unchanged after surfactant instillation, all except one PSR responded with increased total number of impulses and maximal f imp (42 and 26%, respectively). Some HT PSRs ceased to discharge after lung lavage but recovered after surfactant instillation. The end-expiratory activity of LT PSRs increased or was regained after surfactant instillation. After instillation of surfactant, respiratory rate increased further with a shorter inspiratory time, resulting in a lower inspiratory-to-expiratory time ratio. Arterial pH decreased (7.31 +/- 0.04 vs. 7.22 +/- 0.06) and Pco2 increased (5.5 +/- 0.7 vs. 7.2 +/- 1.3 kPa) after lung lavage, but they were the same after as before instillation of surfactant (pH = 7.21 +/- 0.08 and Pco2 = 7.6 +/- 1.4 kPa) during spontaneous breathing. In conclusion, surfactant instillation increased lung compliance, which, in turn, increased the activity of both HT and LT PSRs. A further increase in respiratory rate due to a shorter inspiratory time after surfactant instillation suggests that the partially restored PSR activity after surfactant instillation affected the breathing pattern. PMID- 16210439 TI - Cardiovascular effects of epinephrine during rewarming from hypothermia in an intact animal model. AB - Rewarming from accidental hypothermia is often complicated by "rewarming shock," characterized by low cardiac output (CO) and a sudden fall in peripheral arterial pressure. In this study, we tested whether epinephrine (Epi) is able to prevent rewarming shock when given intravenously during rewarming from experimental hypothermia in doses tested to elevate CO and induce vasodilation, or lack of vasodilation, during normothermia. A rat model designed for circulatory studies during experimental hypothermia and rewarming was used. A total of six groups of animals were used: normothermic groups 1, 2, and 3 for dose-finding studies, and hypothermic groups 4, 5, and 6. At 20 and 24 degrees C during rewarming, group 4 (low-dose Epi) and group 5 (high-dose Epi) received bolus injections of 0.1 and 1.0 microg Epi, respectively. At 28 degrees C, Epi infusion was started in groups 4 and 5 with 0.125 and 1.25 microg/min, respectively. Group 6 served as saline control. After rewarming, both CO and stroke volume were restored in group 4, in contrast to groups 5 and 6, in which both CO and stroke volume remained significantly reduced (30%). Total peripheral resistance was significantly higher in group 5 during rewarming from 24 to 34 degrees C, compared with groups 4 and 6. This study shows that, in contrast to normothermic conditions, Epi infused during hypothermia induces vasoconstriction rather than vasodilation combined with lack of CO elevation. The apparent dissociation between myocardial and vascular responses to Epi at low temperatures may be related to hypothermia induced myocardial failure and changes in temperature-dependent adrenoreceptor affinity. PMID- 16210440 TI - Vasoactive mediators and pulmonary hypertension after cigarette smoke exposure in the guinea pig. AB - The pathogenesis of pulmonary hypertension in patients with chronic obstructive pulmonary disease is not understood. We have previously shown increased levels of mediators that control vasoconstriction (endothelin-1), vascular cell proliferation (endothelin-1 and vascular endothelial growth factor), and vasodilation (endothelial nitric oxide synthase) in the intrapulmonary arteries of animals exposed to cigarette smoke. To determine whether these mediators could be implicated in the structural remodeling of the arterial vasculature and increased pulmonary arterial pressure caused by chronic cigarette smoke exposure, guinea pigs were exposed to daily cigarette smoke for 6 mo. Pulmonary arterial pressures were measured. Intrapulmonary artery structure was analyzed by morphometry, artery mediator protein expression by immunohistochemistry, and artery mediator gene expression by laser capture microdissection and real-time RT PCR. We found that the smoke-exposed animals developed increases in pulmonary arterial pressure and increased muscularization of the small pulmonary arteries. Gene expression and protein levels of all three mediators were increased, and pulmonary arterial pressure correlated both with the levels of mediator production and with the degree of arterial muscularization. We conclude that chronic smoke exposure produces increased vasoactive mediator expression in the small intrapulmonary arteries and that these mediators are associated with vascular remodeling as well as increased pulmonary arterial pressure. These findings support the idea that hypertension in chronic obstructive pulmonary disease is a result of direct cigarette smoke-mediated effects on the vasculature and suggest that interference with endothelin and VEGF production and activity or augmentation of nitric oxide levels may be beneficial. PMID- 16210441 TI - Rehydration with glycerol: endocrine, cardiovascular, and thermoregulatory responses during exercise in the heat. AB - The impact of rehydration with glycerol on cardiovascular and thermoregulatory responses during exercise in the heat was studied in eight highly trained male cyclists. Each subject completed three dehydration-rehydration experimental trials that differed only in the rehydration treatment, each separated by 7 days. Before each experimental day, subjects dehydrated to -4% of their body weight by exercise and water restriction. The experimental treatments were as follows: no fluid (NF), glycerol bolus (1 g/kg body wt) followed by water (G), and water alone (W). Rehydration (3% body weight) was given over an 80-min period. After rehydration, subjects cycled (74% peak O2 uptake) to exhaustion in a hot and wet (37 degrees C and 48% relative humidity) environment. For G, plasma volume was expanded (P < 0.05) during rehydration and remained higher than W (P < 0.05) during exercise. Exercise time to exhaustion during G (33 +/- 4 min) was longer (P < 0.05) compared with both W (27 +/- 3 min) and NF (19 +/- 3 min). Cutaneous vascular conductance was significantly elevated (P < 0.05) during G, but G provided no other thermoregulatory or cardiovascular benefits compared with W and NF. Fluid-regulating hormones (vasopressin, aldosterone, atriopeptin, and plasma renin activity) decreased during rehydration and increased during exercise (except atriopeptin), but there were no differences between G and W. These data indicated that glycerol had little or no major effect on fluid-regulating factors during rehydration or exercise, and the improved exercise capacity in G was likely due to a greater plasma volume during exercise. PMID- 16210442 TI - Low-intensity exercise training during doxorubicin treatment protects against cardiotoxicity. AB - Doxorubicin (Dox) is a highly effective antineoplastic antibiotic associated with a dose-limiting cardiotoxicity that may result in irreversible cardiomyopathy and heart failure. The purpose of this study was to examine the effects of low intensity exercise training (LIET) during the course of Dox treatment on cardiac function, myosin heavy chain expression, oxidative stress, and apoptosis activation following treatment. Male Sprague-Dawley rats either remained sedentary or were exercise trained on a motorized treadmill at 15 m/min, 20 min/day, 5 days/wk (Monday through Friday) for 2 wk. During the same 2-wk period, Dox (2.5 mg/kg) or saline was administered intraperitoneally to sedentary and exercised rats 3 days/wk (Monday, Wednesday, Friday) 1-2 h following the exercise training sessions (cumulative Dox dose: 15 mg/kg). Five days following the final injections, hearts were isolated for determination of left ventricular (LV) function, lipid peroxidation, antioxidant enzyme protein expression, 72-kDa heat shock protein expression, caspase-3 activity, and myosin heavy chain isoform expression. Dox treatment significantly impaired LV function and increased caspase-3 activity in sedentary animals (P < 0.05). LIET attenuated the LV dysfunction and apoptotic signal activation induced by Dox treatment and increased glutathione peroxidase expression, but it had no significant effect on lipid peroxidation, protein expression of myosin heavy chain isoforms, 72-kDa heat shock protein, or superoxide dismutase isoforms. In conclusion, our data suggest that LIET applied during chronic Dox treatment protects against cardiac dysfunction following treatment, possibly by enhancing antioxidant defenses and inhibiting apoptosis. PMID- 16210443 TI - Applicability of segmental bioelectrical impedance analysis for predicting trunk skeletal muscle volume. AB - This study aimed to investigate the validity of using segmental bioelectrical impedance (BI) analysis for estimating skeletal muscle volume (MV) in the trunk, defined as the body segment from the acromion process to the greater trochanter. Using a magnetic resonance imaging (MRI) method, the trunk MV was determined in 28 men (19 approximately 34 yr), divided into validation (n = 20) and cross validation (n = 8) groups, and used as a reference (MV(MRI)). For BI measurements of the trunk, the source electrodes were placed at the dorsal surface of the third metacarpal bone of both hands and the dorsal surface of the third metatarsal bone of both feet, and the detector electrodes were placed at the acromion process of both shoulders and the greater trochanter of both femurs. Using this arrangement, the BI values of five parts of the trunk, both sides of the upper region, the middle region, and both sides of the lower region, were obtained and then used to calculate the whole trunk BI value and BI index (BI index(TR)). In the validation group, a simple regression analysis of the relationship between BI index(TR) and MV(MRI) showed a significant correlation between the two variables (r = 0.884, P < 0.05) and produced a prediction equation with a SE of estimation of 1,020.3 cm3 (8.5%). In the validation and cross-validation groups, there were no significant differences between the measured and estimated MV without systematic errors. These findings indicate that the segmental BI analysis employed in the present study can be used to estimate trunk MV. PMID- 16210444 TI - Pulmonary ischemia induces lung remodeling and angiogenesis. AB - Cellular remodeling during angiogenesis in the lung is poorly described. Furthermore, it is the systemic vasculature of the lung and surrounding the lung that is proangiogenic when the pulmonary circulation becomes impaired. In a mouse model of chronic pulmonary thromboembolism, after left pulmonary artery ligation (LPAL), the intercostal vasculature, in proximity to the ischemic lung, proliferates and invades the lung (12). In the present study, we performed a detailed investigation of the kinetics of remodeling using histological sections of the left lung of C57Bl/6J mice after LPAL (4 h to 20 days) or after sham surgery. New vessels were seen within the thickened visceral pleura 4 days after LPAL predominantly in the upper portion of the left lung. Connections between new vessels within the pleura and pulmonary capillaries were clearly discerned by 7 days after LPAL. The visceral pleura and the lung parenchyma showed intense tissue remodeling, as evidenced by markedly elevated levels of both proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling positive cells. Rapidly dividing cells were predominantly macrophages and type II pneumocytes. The increased apoptotic activity was further quantified by caspase-3 activity, which showed a sixfold increase relative to naive lungs, by 24 h after LPAL. Because sham surgeries had little effect on measured parameters, we conclude that both thoracic wound healing and pulmonary ischemia are required for systemic neovascularization. PMID- 16210445 TI - Examining axial diffusion of nitric oxide in the lungs using heliox and breath hold. AB - Exhaled nitric oxide (NO) is highly dependent on exhalation flow; thus exchange dynamics of NO have been described by multicompartment models and a series of flow-independent parameters that describe airway and alveolar exchange. Because the flow-independent NO airway parameters characterize features of the airway tissue (e.g., wall concentration), they should also be independent of the physical properties of the insufflating gas. We measured the total mass of NO exhaled (A(I,II)) from the airways after five different breath-hold times (5-30 s) in healthy adults (21-38 yr, n = 9) using air and heliox as the insufflating gas, and then modeled A(I,II) as a function of breath-hold time to determine airway NO exchange parameters. Increasing breath-hold time results in an increase in A(I,II) for both air and heliox, but A(I,II) is reduced by a mean (SD) of 31% (SD 6) (P < 0.04) in the presence of heliox, independent of breath-hold time. However, mean (SD) values (air, heliox) for the airway wall diffusing capacity [3.70 (SD 4.18), 3.56 pl.s(-1).ppb(-1) (SD 3.20)], the airway wall concentration [1,439 (SD 487), 1,503 ppb (SD 644>)], and the maximum airway wall flux [4,156 (SD 2,502), 4,412 pl/s (SD 2,906)] using a single-path trumpet-shaped airway model that considers axial diffusion were independent of the insufflating gas (P > 0.55). We conclude that a single-path trumpet model that considers axial diffusion captures the essential features of airway wall NO exchange and confirm earlier reports that the airway wall concentration in healthy adults exceeds 1 ppm and thus approaches physiological concentrations capable of modulating smooth muscle tone. PMID- 16210446 TI - Arterial baroreflex resetting during exercise: a current perspective. AB - Within the past 20 years numerous animal and human experiments have provided supportive evidence of arterial baroreflex resetting during exercise. In addition, it has been demonstrated that both the feedforward mechanism of central command and the feedback mechanism associated with skeletal muscle afferents (the exercise pressor reflex) play both independent and interactive roles in the resetting of the arterial baroreflex with exercise. A fundamental alteration associated with baroreflex resetting during exercise is the movement of the operating point of the reflex away from the centring point and closer to the threshold, thereby increasing the ability of the reflex to buffer hypertensive stimuli. Recent studies suggest that central command and the cardiopulmonary baroreceptors may play a role in this movement of the operating point on the baroreflex-heart rate and baroreflex-blood pressure curve, respectively. Current research is focusing on the investigation of central neural mechanisms involved in cardiovascular control, including use of electrophysiological and molecular biological techniques in rat and mouse models to investigate baroreflex resetting as well as use of state of the art brain imaging techniques in humans. However, the purpose of this review is to describe the role of the arterial baroreflex in the regulation of arterial blood pressure during physical activity from a historical perspective with a particular emphasis on human investigations. PMID- 16210447 TI - Maximal motor unit firing rates during isometric resistance training in men. AB - This study measured changes in maximal voluntary contraction (MVC) force, percentage maximal activation, maximal surface EMG, M-wave amplitude and average motor unit firing rates during the initial 3 weeks of isometric resistance training of the quadriceps muscle. Ten men participated in a resistance training programme three times a week for 3 weeks and 10 men participated as a control group. In the training group, MVC increased by 35% (from 761 +/- 77 to 1031 +/- 78 N) by the end of the 3 weeks. There were no changes in mean motor unit firing rates during submaximal or maximal voluntary contractions of 50 (15.51 +/- 1.48 Hz), 75 (20.23 +/- 1.85 Hz) or 100% MVC (42.25 +/- 2.72 Hz) with isometric resistance training. There was also no change in maximal surface EMG relative to the M-wave amplitude. However, there was a small increase in maximal activation (from 95.7 +/- 1.83 to 98.44 +/- 0.66%) as measured by the twitch interpolation technique. There were no changes in any of the parameters measured in the control group. It is suggested that mechanisms other than increases in average motor unit firing rates contributed to the increase in maximal force output with resistance training. Such mechanisms may include a combination of increased motor unit recruitment, enhanced protein synthesis, and changes in motor unit synchronization and muscle activation patterns across the quadriceps synergy. PMID- 16210448 TI - Effects of rhythmic muscle compression on cardiovascular responses and muscle oxygenation at rest and during dynamic exercise. AB - We examined the way in which the duration of rhythmic muscle compressions affects cardiovascular responses and muscle oxygenation at rest and during dynamic exercise. We measured the mean arterial pressure (MAP), heart rate (HR) and oxygenation of the vastus lateralis muscle (by near-infrared spectroscopy) in eight healthy male subjects at rest and during supine bicycle exercise (50 and 100 W at 60 r.p.m.) while applying pulsed muscle compressions at 1000 ms intervals. Compression pressure and durations were 150 mmHg and 300, 600, 900 and 1000 ms (1000 ms being static continuous compression), respectively. During exercise, the pulsed leg compression was synchronized to each thigh extensor muscle contraction. The observed changes in muscle oxygenation were dependent on compression duration (increased at 300 ms, no change at 600 ms and decreased at 900 or 1000 ms) and were different from those seen at rest (increases at < 1000 ms and decrease at 1000 ms). This suggests that the effects of external pulsed muscle compression may have a duration threshold below which muscle pumping counteracts the obstruction to flow caused by the compression, and that the threshold is set at a shorter compression duration during exercise than at rest. Although HR and MAP did not change during pulsed compression at rest, during exercise they both increased progressively as compression duration increased. Thus, while exercising, the increased MAP and HR seen during the compression could be due to the combination and interaction of mechanical effects and the muscle mechanoreflex and/or metaboreflex. PMID- 16210449 TI - Renal sympathetic and circulatory responses to activation of the exercise pressor reflex in rats. AB - We investigated the role played by the exercise pressor reflex in sympathetic regulation of the renal circulation in rats. In mid-collicular decerebrate rats, mean arterial pressure (MAP), heart rate (HR), left renal cortical blood flow (RCBF) and left renal sympathetic nerve activity (RSNA) were recorded before and during 30 s of static contraction of the left triceps surae muscles evoked by electrical stimulation of the tibial nerve, which activates both metabo- and mechanosensitive muscle afferents, and during 30 s of passive stretch of the left Achilles tendon, which selectively activates mechanosensitive muscle afferents. Static contraction (n = 17, +344 +/- 34 g developed tension) significantly (P < 0.05) increased MAP (+14 +/- 3 mmHg), HR (+6 +/- 1 beats min(-1)) and RSNA (n = 11, +19 +/- 5%) and significantly decreased renal cortical vascular conductance (RCVC, n = 11, -11 +/- 2%). Passive stretch (n = 20, +378 +/- 11 g) also significantly increased MAP (+11 +/- 2 mmHg), HR (+7 +/- 2 beats min(-1)) and RSNA (n = 15, +14 +/- 4%) and significantly decreased RCVC (n = 11, -12 +/- 3%). RCBF showed no significant changes during static contraction or passive stretch. Renal denervation abolished the decrease in RCVC during contraction (n = 12) or stretch (n = 13). These data indicate that both the exercise pressor reflex and its mechanically sensitive component, the muscle mechanoreflex, induced renal cortical vasoconstriction through sympathetic activation in rats. PMID- 16210450 TI - Organ-specific ligation-induced changes in harmonic components of the pulse spectrum and regional vasoconstrictor selectivity in Wistar rats. AB - It has been shown previously that the amplitudes of the harmonic components of the pulse spectrum vary in specific patterns when the arteries leading to different organs are ligated, with the variations in the harmonics being linearly additive. Since ligation can be regarded as a vast increase in organ resistance, the present study examined the potential of using these ligation-induced variations in the pulse spectrum as reference parameters for an increase in vascular resistance and for regional vasoconstrictor selectivity. A vasoconstrictor, either arginine vasopressin (AVP) or angiotensin II (Ang II), was infused into anaesthetized Wistar rats via the femoral vein for 1 h. The distinct harmonic-specific drug effects on the pulse spectrum were simulated by combining renal artery and superior mesenteric artery ligations in different ratios, the ratio with the lowest mean square difference determining the regional drug selectivity. The ratios indicated that the effect of AVP on the pulse spectrum was attributable to the combined effect of ligating the renal and superior mesenteric arteries, while the effect of Ang II was attributable to ligation of the renal artery. The results are comparable with those of investigations of regional vascular resistance performed using traditional methods. Our findings indicate that the ligation-induced variations in the pulse spectrum can be used to determine regional increases in vascular resistance. This implies that blood pressure can be used as the sole parameter to determine which arterial bed has been affected by the vasoconstrictor, and how seriously. PMID- 16210451 TI - Mineralocorticoids decrease the activity of the apical small-conductance K channel in the cortical collecting duct. AB - We used the patch-clamp technique to examine the effect of DOCA treatment (2 mg/kg) on the apical small-conductance K (SK) channels, epithelial Na channels (ENaC), and the basolateral 18-pS K channels in the cortical collecting duct (CCD). Treatment of rats with DOCA for 6 days significantly decreased the plasma K from 3.8 to 3.1 meq and reduced the activity of the SK channel, defined as NP(o), from 1.3 in the CCD of control rats to 0.6. In contrast, DOCA treatment significantly increased ENaC activity from 0.01 to 0.53 and the basolateral 18-pS K channel activity from 0.67 to 1.63. Moreover, Western blot analysis revealed that DOCA treatment significantly increased the expression of the nonreceptor type of protein tyrosine kinase (PTK), cSrc, and the tyrosine phosphorylation of ROMK in the renal cortex and outer medulla. The possibility that decreases in apical SK channel activity induced by DOCA treatment were the result of stimulation of PTK activity was further supported by experiments in which inhibition of PTK with herbimycin A significantly increased NP(o) from 0.6 to 2.1 in the CCD from rats receiving DOCA. Also, when rats were fed a high-K (10%) diet, DOCA treatment did not increase the expression of c-Src and decrease the activity of the SK channel in the CCD. We conclude that DOCA treatment decreased the apical SK channel activity in rats on a normal-K diet and that an increase in PTK expression may be responsible for decreased channel activity in the CCD from DOCA-treated rats. PMID- 16210452 TI - Testosterone supplementation in aging men and women: possible impact on cardiovascular-renal disease. AB - Treatment of aging men and women with testosterone supplements is increasing. The supplements are given to postmenopausal women mainly to improve their libido and to aging men to improve muscle mass and bone strength, to improve libido and quality of life, to prevent and treat osteoporosis, and, with the phosphodiesterase-5 inhibitors, such as sildenafil, to treat erectile dysfunction. The increased use of testosterone supplements in aging individuals has occurred despite the fact that there have been no rigorous clinical trials examining the effects of chronic testosterone on the cardiovascular-renal disease risk. Studies in humans and animals have suggested that androgens can increase blood pressure and compromise renal function. Androgens have been shown to increase tubular sodium and water reabsorption and activate various vasoconstrictor systems in the kidney, such as the renin-angiotensin system and endothelin. There is also evidence that androgens may increase oxidative stress. Furthermore, the kidney contains the enzymes necessary to produce androgens de novo. This review presents an overview of the data from human and animal studies in which the role of androgens in promoting renal and cardiovascular diseases has been investigated. PMID- 16210453 TI - Expression and functions of annexins in the kidney. AB - This review article summarizes current knowledge about the locations and possible functions of annexin family members in the kidney. Beginning with an introduction on common structural and biochemical features as well as general functional characteristics of annexins, the paper focuses on individual members with documented and/or proposed physiological relevance for renal development, structure, and functions. Three main aspects of annexin function in kidney epithelia emerge from the available experimental data. First, annexins are required for membrane organization and membrane transport events required for the establishment/maintenance of epithelial polarity. Second, there is accumulating evidence of an association of annexins with ion channels, as membrane-guiding auxiliary proteins or modulators of channel activity. Last but not least, some annexins seem to work as extracellular autocrine modulators of receptor function under different physiological conditions. PMID- 16210454 TI - Recurrent haemoptysis with anaemia in a 16 year old man. PMID- 16210455 TI - HIV/AIDS prevention in Uganda: why has it worked? PMID- 16210456 TI - Toward health and wellbeing for indigenous Australians. AB - The health of indigenous Australians remains well below that of non-indigenous Australians and indigenous peoples in Canada and New Zealand. Although recent planning has initiated many outstanding, culturally appropriate programmes with indigenous involvement, health statistics only reflect marginal improvement in recent years. It is crucial that positive programmes are sustained with appropriately directed funding. An approach that includes respect for the emotional and spiritual wellbeing of Australia's indigenous peoples will assist to redress some of the disadvantage caused by dispossession of country, language, and identity. It is clear from many programmes that are in place, that primary health care delivered locally through community controlled organisations, will minimise the impact of serious illnesses that currently threaten whole families and communities. Westernized health care systems are slow to learn from indigenous peoples in Australia and other places, that maintenance of wellness, not management of illness should be the goal. PMID- 16210457 TI - Revisiting the ABC strategy: HIV prevention in Uganda in the era of antiretroviral therapy. AB - The ABC strategy is credited for bringing the HIV/AIDS epidemic under control in Uganda. By promoting abstinence, being faithful, and condom use, safe(r) behaviours have been identified that are applicable to people in different circumstances. However, scaling-up of antiretroviral therapy in the country raised concerns that HIV prevention messages targeting the uninfected population are not taking sufficient account of inherent complexities. Furthermore, there is debate in the country over relative importance of abstinence in reduction of HIV incidence as well as over the morality and effectiveness of condoms. The purpose of this paper is to examine each component of ABC in light of current developments. It is argued that there is still a strong justification for condom use to complement abstinence and being faithful. There is an urgent need to update and relaunch Uganda's ABC strategy--its three elements are complementary, synergistic, and inseparable in the national HIV prevention programme. PMID- 16210458 TI - Nutrition support for patients in the intensive care unit. AB - Enteral nutrition (EN) is the mainstay of nutrition delivery within intensive care seeking to capitalise on its benefits for the gastrointestinal tract and associated immune system, but this has brought new challenges in delivery to the sick. The hoped for benefit has led to the mistaken belief by some that parenteral nutrition (PN) is no longer required. However, a greater appreciation of the risks of EN delivery in the sick patient combined with improvements in PN formulation and use help explain why PN is not as risky as some have believed. Real outcome benefits have been described with the new glutamine containing PN formulations. PN remains important in the presence of gastrointestinal feed intolerance or failure. PMID- 16210459 TI - Non-invasive ventilation in acute cardiogenic pulmonary oedema. AB - Non-invasive ventilation (NIV) is the delivery of assisted mechanical ventilation to the lungs, without the use of an invasive endotracheal airway. NIV has revolutionised the management of patients with various forms of respiratory failure. It has decreased the need for invasive mechanical ventilation and its attendant complications. Cardiogenic pulmonary oedema (CPO) is a common medical emergency, and NIV has been shown to improve both physiological and clinical outcomes. From the data presented herein, it is clear that there is sufficiently high level evidence to favour the use of continuous positive airway pressure (CPAP), and that the use of CPAP in patients with CPO decreases intubation rate and improves survival (number needed to treat seven and eight respectively). However, there is insufficient evidence to recommend the use of bilevel positive airway pressure (BiPAP), probably the exception being patients with hypercapnic CPO. More trials are required to conclusively define the role of BiPAP in CPO. PMID- 16210460 TI - Psychosis in Parkinson's disease. AB - Psychosis is common in Parkinson's disease (PD), particularly in its later stages. The symptoms range from comparatively minor illusions, vivid dreams, and occasional, non-disturbing visual hallucinations to frank psychosis. The pathogenesis of psychosis in PD is not fully known. Management of psychosis in PD requires a multidisciplinary approach. Some of the newer atypical antipsychotics are effective against psychosis with no significant worsening of PD. Psychosis in PD is associated with poor quality of life for patients and the carers. PMID- 16210461 TI - Monitoring surgical and medical outcomes: the Bernoulli cumulative SUM chart. A novel application to assess clinical interventions. AB - BACKGROUND: Monitoring clinical interventions is an increasing requirement in current clinical practice. The standard CUSUM (cumulative sum) charts are used for this purpose. However, they are difficult to use in terms of identifying the point at which outcomes begin to be outside recommended limits. OBJECTIVE: To assess the Bernoulli CUSUM chart that permits not only a 100% inspection rate, but also the setting of average expected outcomes, maximum deviations from these, and false positive rates for the alarm signal to trigger. METHODS: As a working example this study used 674 consecutive first liver transplant recipients. The expected one year mortality set at 24% from the European Liver Transplant Registry average. A standard CUSUM was compared with Bernoulli CUSUM: the control value mortality was therefore 24%, maximum accepted mortality 30%, and average number of observations to signal was 500-that is, likelihood of false positive alarm was 1:500. RESULTS: The standard CUSUM showed an initial descending curve (nadir at patient 215) then progressively ascended indicating better performance. The Bernoulli CUSUM gave three alarm signals initially, with easily recognised breaks in the curve. There were no alarms signals after patient 143 indicating satisfactory performance within the criteria set. CONCLUSIONS: The Bernoulli CUSUM is more easily interpretable graphically and is more suitable for monitoring outcomes than the standard CUSUM chart. It only requires three parameters to be set to monitor any clinical. INTERVENTION: the average expected outcome, the maximum deviation from this, and the rate of false positive alarm triggers. PMID- 16210462 TI - An investigation of the anger levels of residents: medical compared with surgical disciplines. AB - OBJECTIVE: To evaluate medical and surgical residents' anger levels with regard to the department in which they worked, seniority, sex, satisfaction with their work environment, and the number of nightshifts worked per month. The specific situations and persons at whom residents reacted with anger were also investigated. METHODS: 116 randomly selected residents staffed in a university hospital (62 medical and 54 surgical residents) were enrolled. The trait anger and anger expression scale was used to find out the personal anger levels of each participant. The participants also clarified the persons and situations that made them angry at work. RESULTS: Trait anger levels were greater in the surgical residents in their first two years when compared with levels of their senior colleagues (p = 0.033). Mean trait anger levels were greater in the residents who were not satisfied with their department (p = 0.004). Anger levels were not found to be related to the number of shifts per month. Male residents had higher levels of anger than female colleagues (p = 0.019). CONCLUSION: Residents in clinical sciences seem to have the potential to benefit from a screening process in terms of anger and its subcomponents by means of a tool such as the trait anger and anger expression scale during their residency. PMID- 16210463 TI - Correlation between clinical diagnosis and arthroscopic findings of the shoulder. AB - OBJECTIVE: To assess the accuracy of clinical examination by non-specialist orthopaedic surgeons of patients presenting to a diagnostic and treatment centre (DTC) for arthroscopic shoulder surgery. METHODS: A retrospective review of notes of 130 consecutive shoulder arthroscopies performed at a DTC over a 10 month period. Preoperative clinical diagnosis was compared with operative arthroscopic findings. Additional information from preoperative imaging was compared with clinical examination and arthroscopic findings. Preoperative clinical examinations and consent were undertaken by clinical fellows, (SpR level) and non upper limb consultant orthopaedic surgeons. Consultants specialising in upper limb surgery performed the operations. RESULTS: Six main groups were identified on the basis of clinical examination: impingement 76 cases (58%), instability 22 cases (17%), frozen shoulder 11 cases (8%), rotator cuff tear four cases (3%), non-specific pain eight cases (6%), and normal clinical examination nine cases (7%). Impingement and instability diagnosed clinically strongly correlated with the arthroscopic findings. Clinical diagnosis of frozen shoulder and rotator cuff tears had a weaker correlation with the arthroscopic findings. Of the nine cases of normal clinical examination, abnormality was found at arthroscopy in all cases. CONCLUSION: There have been very few studies comparing clinical examination of the shoulder with arthroscopic findings. This study emphasises the importance of good clinical examination skills in diagnosing common shoulder abnormalities. The addition of imaging, particularly ultrasound and magnetic resonance imaging further increases the likelihood of an accurate diagnosis. Shoulder examination should be taught with as much emphasis at both undergraduate and postgraduate level as other orthopaedic clinical examinations. PMID- 16210465 TI - Radiation exposure to personnel performing endoscopic retrograde cholangiopancreatography. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography (ERCP) relies on the use of ionising radiation but risks to operator and patient associated with radiation exposure are unclear. The aim of this prospective study was to estimate the radiation dose received by personnel performing fluoroscopic endoscopic procedures, mainly ERCP. METHODS: Consecutive procedures over a two month period were included. The use of thermoluminescent dosimeters to measure radiation exposure to the abdomen, thyroid gland, and hands of the operator permitted an estimation of the annual whole body effective dose equivalent. RESULTS: During the study period 66 procedures (61 ERCP) were performed and the estimated annual whole body effective dose equivalent received by consultant operators ranged between 3.35 and 5.87 mSv. These values are similar to those received by patients undergoing barium studies and equate to an estimated additional lifetime fatal cancer risk between 1 in 7000 and 1 in 3500. While within legal safety limits for radiation exposure to personnel, these doses are higher than values deemed acceptable for the general public. CONCLUSIONS: It is suggested that personnel as well as patients may be exposed to significant values of radiation during ERCP. The study emphasises the need to carefully assess the indication for, and to use measures that minimise radiation exposure during any fluoroscopic procedure. PMID- 16210466 TI - The "OBS" chart: an evidence based approach to re-design of the patient observation chart in a district general hospital setting. AB - OBJECTIVE: The primary role of the patient bedside observation chart is to make clinicians aware of the deteriorating patient. Despite this, its performance has not been scrutinised. Many versions exist with different styles of data entry but the optimal format remains elusive. This paper hypothesised that chart design measurably influences function and that redesign and standardisation would improve the detection of physiological decline by clinical staff. DESIGN: Objective evaluation of existing charts (n = 5), evidence based redesign, and re evaluation of new chart. SETTING: 250 bed district general hospital. RESULTS: Design of existing observation charts had a significant effect on the ability of clinical staff to detect patient deterioration, with detection rates of parameters indicating physiological decline ranging from 0% to 100%. Graphical plots portrayed information better than written values for all parameters being measured except tachypnoea. No single existing chart was best for all variables. A new chart was designed, implemented with training in its use, and re-evaluated. The new chart also incorporated an early warning scoring system. There were significant improvements in the average detection rates of parameters poorly identified on existing charts: detection rates of tachypnoea and hypoxia increased by 41% (p<0.05) and 45% (p<0.05) respectively. There were also significant improvements in detection rates of tachycardia and fever by 29% (p<0.05) and 16% (p<0.05) respectively. CONCLUSION: Evidence based redesign of the patient bedside observation chart coupled with specific training in its use significantly improves the detection of patient physiological deterioration. PMID- 16210467 TI - Change of nitric oxide in experimental colitis and its inhibition by melatonin in vivo and in vitro. AB - AIM: To investigate the change of nitric oxide (NO) in rat colitis and its inhibition by melatonin in vivo and in vitro. METHODS: In vivo, rat colitis was established intracolonically with trinitrobenzenesulphonic acid (TNBS) and ethanol. The animals were randomised into five groups: control group, model group, melatonin group (2.5, 5.0, 10.0 mg/kg), and treated intracolonically with saline, saline and melatonin respectively (once a day, from day 7 after colitis was established to day 28). After the end of the experiment, the mucosal damage index (CMDI) and histology score (HS) were evaluated and the level of myeloperoxidase (MPO) and malondiadehyde (MDA) and NO in the colon tissue were measured. In vitro, the co-culture model of the inflamed colon mucosa (from the colitis) with lipopolysaccharide (LPS), and the colonocytes oxidative injury model by hydroxyl radical, were designed respectively to elucidate the inhibition of NO by melatonin. RESULTS: After treated with TNBS/ethanol, the extent of CMDI and HS, the levels of MPO, MDA, and NO in the model group, were higher than that in the control group; melatonin ameliorated these parameters effectively. The stimulation of LPS increased the level of NO and MPO and MDA in the co-culture model of inflamed colon mucosa, and melatonin significantly reduced the level of MPO, MDA, and NO. In the coloncyte oxidative injury model by hydroxyl radical, the contents of LDH, MDA, and NO were increased; melatonin reversed this oxidative injury considerably. CONCLUSION: This study showed that TNBS/ethanol induced colitis was pharmacologically controlled by melatonin in vivo and in vitro. PMID- 16210469 TI - Durability of marker-quantitative trait loci haplotypes in structured populations. AB - Given the relative ease of identifying genetic markers linked to QTL (compared to finding the loci themselves), it is natural to ask whether linked markers can be used to address questions concerning the contemporary dynamics and recent history of the QTL. In particular, can a marker allele found associated with a QTL allele in a QTL mapping study be used to track population dynamics or the history of the QTL allele? For this strategy to succeed, the marker-QTL haplotype must persist in the face of recombination over the relevant time frame. Here we investigate the dynamics of marker-QTL haplotype frequencies under recombination, population structure, and divergent selection to assess the potential utility of linked markers for a population genetic study of QTL. For two scenarios, described as "secondary contact" and "novel allele," we use both deterministic and stochastic methods to describe the influence of gene flow between habitats, the strength of divergent selection, and the genetic distance between a marker and the QTL on the persistence of marker-QTL haplotypes. We find that for most reasonable values of selection on a locus (s < or = 0.5) and migration (m > 1%) between differentially selected populations, haplotypes of typically spaced markers (5 cM) and QTL do not persist long enough (>100 generations) to provide accurate inference of the allelic state at the QTL. PMID- 16210471 TI - Albumin turnover: FcRn-mediated recycling saves as much albumin from degradation as the liver produces. AB - It is now understood that the nonclassical major histocompatibility complex-I molecule FcRn binds albumin and retrieves it from an intracellular degradative fate. Whether FcRn in the liver modulates albumin turnover through effects on biosynthesis and production is not known. Thus we quantified the appearance of biosynthetically labeled albumin in plasma after an intravenous bolus injection of [(3)H]leucine in FcRn-deficient mice. The production rates for both albumin (FcRn substrate) and transferrin (nonsubstrate) are increased by approximately 20% in FcRn-deficient mice compared with normal mice, likely compensating for the lowered plasma oncotic pressure caused by hypoalbuminemia in FcRn-deficient mice. Determining the magnitude of FcRn-mediated effects on albumin turnover, we then measured the steady-state plasma concentrations of biosynthetically labeled albumin and transferrin during [(3)H]leucine infusion. The concentration of albumin was approximately 40% lower in FcRn-deficient mice compared with normal mice. Furthermore, the approximately 40% lower plasma albumin concentration in FcRn-deficient mice along with the approximately 20% increase in albumin production indicate, by the mass-balance equation, that albumin degradation in FcRn-deficient mice is twice that of normal mice. These studies of biosynthetically labeled, and thus native, albumin support our previous finding that FcRn protects albumin from degradation. Permitting quantification of the magnitude of FcRn-mediated recycling, they further indicate that FcRn has extraordinary capacity: the amount of albumin saved from degradation by FcRn mediated recycling is the same as that produced by the liver. PMID- 16210472 TI - Synchrony between circular and longitudinal muscle contractions during peristalsis in normal subjects. AB - The current understanding is that longitudinal muscle contraction begins before and outlasts circular muscle contraction during esophageal peristalsis in normal subjects. The goal of our study was to reassess the relationship between the contractility of two muscle layers using novel ways to look at the muscle contraction. We studied normal subjects using synchronized high-frequency ultrasound imaging and manometry. Swallow-induced peristalsis was recorded at 5 and 10 cm above the lower esophageal sphincter (LES). Ultrasound (US) images were analyzed for muscle cross-sectional area (CSA) and circularity index of the esophagus during various phases of esophageal contraction. A plot of the M mode US image, muscle CSA, and esophageal circularity index was developed to assess the temporal correlation between various parameters. The muscle CSA wave began before and lasted longer than the contraction pressure wave at both 5 and 10 cm above the LES. M mode US images revealed that the onset of muscle CSA wave was temporally aligned with the onset of lumen collapse. The peak muscle CSA occurred in close proximity with the peak pressure wave. The esophagus started to become more circular (decrease in circularity index) with the onset of the muscle CSA wave. The circularity index and muscle CSA returned to the baseline at approximately the same time. In conclusion, the onset of lumen collapse and return of circularity index of the esophagus are likely to be the true markers of the onset and end of circular muscle contraction. Circular and longitudinal muscle layers of the esophagus contract in a precise synchronous fashion during peristalsis in normal subjects. PMID- 16210470 TI - EGF-mediated regulation of IGFBP-3 determines esophageal epithelial cellular response to IGF-I. AB - IGF and EGF regulate various physiological and pathological processes. IGF binding protein (IGFBP)-3 regulates cell proliferation in IGF-dependent and independent fashions. Recently, we identified IGFBP-3 as a novel EGF receptor (EGFR) downstream target molecule in primary and immortalized human esophageal epithelial cells, suggesting an interplay between the EGF and IGF signaling pathways. However, the regulatory mechanisms for IGFBP-3 expression and its functional role in esophageal cell proliferation remain to be elucidated. Herein, we report that IGFBP-3 mRNA and protein were induced upon growth factor deprivation in primary and immortalized human esophageal cells through mechanisms requiring p53-independent de novo mRNA transcription and protein synthesis. This occurred in the face of the activated phosphatidylinositol 3-OH-kinase (PI3K)/mammalian target of rapamycin (mTOR) pathway. Secreted IGFBP-3 neutralized IGFs and prevented IGF-I receptor (IGF-IR) activation. In contrast, EGF suppressed IGFBP-3 mRNA and protein expression through activation of MAPK in an EGFR-tyrosine kinase-dependent manner to restore the cellular response to IGF-I. When stably overexpressed, wild-type IGFBP-3 but not I56G/L80G/L81G (GGG) mutant IGFBP-3, which has a reduced affinity to IGFs, prevented IGF-I from activating IGF-IR and Akt as well as stimulating cell proliferation. However, unlike other cell types where IGFBP-3 exerts antiproliferative effects, neither wild-type nor GGG mutant IGFBP-3 alone affected cell proliferation or EGFR activity. These results indicate that IGF signaling is subject to negative regulation through IGFBP-3 and positive regulation by EGF, the latter of which suppresses IGFBP-3. This provides a platform for understanding the novel cross talk between EGF- and IGF-mediated pathways. PMID- 16210473 TI - Adaptor heat shock protein complex formation regulates trafficking of the asialoglycoprotein receptor. AB - In the asialoglycoprotein receptor (ASGPR) endocytic pathway, internalized receptors pass through early, recycling, and sorting endosomal compartments before returning to the cell surface. Sorting motifs in the cytoplasmic domain (CD) and protein interactions with these sequences presumably direct receptor trafficking. Previous studies have shown that association of a potential sorting heat shock protein (HSP) heterocomplex with the ASGPR-CD was regulated by casein kinase 2 (CK2)-mediated phosphorylation. Mass spectrometry and immunoblot analyses identified five of these ASGPR-CD-associated proteins as the molecular chaperones glycoprotein 96, HSP70, HSP90, cyclophilin A, and FK 506 binding protein. The present study was undertaken to determine whether any of the adaptor protein complexes (AP1, AP2, or AP3) were selectivity associated with the ASGPR CD. In conjunction with molecular chaperones, AP2 and AP1 were recovered from a CK2 phosphorylated agarose-GSH-GST-ASGPR-CD matrix. Binding of AP3 was independent of the phosphorylation status of the CD matrix. Inhibition of CK2 mediated phosphorylation with tetrabromobenzotriazole prevented AP recovery within an immunoadsorbed ASGPR complex. Rapamycin, which dissociates the HSP heterocomplex from ASGPR-CD, thereby altering receptor trafficking also, inhibited AP association. Similar results were obtained with an inhibitor of HSP90 heterocomplex formation, geldanmycin. The data presented provide evidence that recruitment of AP1 and AP2, which is necessary for appropriate receptor trafficking, is mediated by the interaction of AP with the ASGPR-CD-bound HSP complex. PMID- 16210475 TI - Development of a microsphere-based serologic multiplexed fluorescent immunoassay and a reverse transcriptase PCR assay to detect murine norovirus 1 infection in mice. AB - Murine norovirus 1 (MNV-1) is a newly recognized pathogen of mice that causes lethal infection in mice deficient in components of the innate immune response but not in wild-type 129 mice. In this study, in vitro-propagated MNV-1 was used as antigen to develop a multiplexed fluorescent immunoassay (MFI) to detect antibodies to MNV-1 in infected mice. The MNV-1 MFI was 100% specific and 100% sensitive in detecting anti-MNV-1 antibody in sera from experimentally infected mice. Testing of a large number of mouse serum samples (n = 12,639) submitted from contemporary laboratory mouse colonies in the United States and Canada revealed that 22.1% of these sera contained antibodies to MNV-1, indicating infection with MNV-1 is widespread in research mice. In addition, a reverse transcriptase PCR primer pair with a sensitivity of 25 virus copies was developed and used to demonstrate that MNV-1 RNA could be detected in the spleen, mesenteric lymph node, and jejunum from some experimentally infected mice 5 weeks postinoculation. These diagnostic assays provide the necessary tools to define the MNV-1 infection status of research mice and to aid in the establishment of laboratory mouse colonies free of MNV-1 infection. PMID- 16210474 TI - Sensitivity and specificity of a monoclonal antibody-based fluorescence assay for detecting Enterocytozoon bieneusi spores in feces of simian immunodeficiency virus-infected macaques. AB - Enterocytozoon bieneusi is clinically the most significant among the microsporidia causing chronic diarrhea, wasting, and cholangitis in individuals with human immunodeficiency virus/AIDS. Microscopy with either calcofluor or modified trichrome stains is the standard diagnostic test for microsporidiosis and does not allow species identification. Detection of E. bieneusi infection based on PCR is limited to a few reference laboratories, and thus it is not the standard diagnostic assay. We have recently reported the development and characterization of a panel of monoclonal antibodies against E. bieneusi, and in this publication we evaluated the specificity and sensitivity of an immunofluorescence assay (IFA), compared with PCR, in simian immunodeficiency virus-infected macaques. The IFA, which correlated with the primary PCR method, with a detection limit of 1.5 x 10(5) spores per gram of feces, will simplify considerably the detection of E. bieneusi spores in clinical and environmental specimens and in laboratory and epidemiological investigations. PMID- 16210476 TI - Evaluation of immunoglobulin G subclass antibodies against recombinant Fasciola gigantica cathepsin L1 in an enzyme-linked immunosorbent assay for serodiagnosis of human fasciolosis. AB - A cystatin capture enzyme-linked immunosorbent assay (ELISA) using recombinant Fasciola gigantica cathepsin L1 antigen was developed to detect specific immunoglobulin G (IgG) subclass antibodies (IgG1, IgG2, IgG3, and IgG4) and was evaluated for its diagnostic potential for human fasciolosis. In an analysis of the sera of 13 patients infected with F. gigantica, 209 patients with other parasitic infections, 32 cholangiocarcinoma patients, and 42 healthy controls, the IgG4-ELISA gave the highest diagnostic values. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method based on the detection of IgG4 antibody were 100%, 99.3%, 99.3%, 86.7%, and 100%, respectively. The results revealed that restricting the ELISA to the detection of specific IgG4 antibody enhanced the specificity and accuracy for the serodiagnosis of human fasciolosis. PMID- 16210477 TI - Evaluation of serum antibody responses against the rotavirus nonstructural protein NSP4 in children after rhesus rotavirus tetravalent vaccination or natural infection. AB - The immune response elicited by the rotavirus nonstructural protein NSP4 and its potential role in protection against rotavirus disease are not well understood. We investigated the serological response to NSP4 and its correlation with disease protection in sera from 110 children suffering acute diarrhea, associated or not with rotavirus, and from 26 children who were recipients of the rhesus rotavirus tetravalent (RRV-TV) vaccine. We used, as antigens in an enzyme-linked immunosorbent assay (ELISA), affinity-purified recombinant NSP4 (residues 85 to 175) from strains SA11, Wa, and RRV (genotypes A, B, and C, respectively) fused to glutathione S-transferase. Seroconversion to NSP4 was observed in 54% (42/78) of the children who suffered from natural rotavirus infection and in 8% (2/26) of the RRV-TV vaccine recipients. Our findings indicate that NSP4 evokes significantly (P < 0.05) higher seroconversion rates after natural infection than after RRV-TV vaccination. The serum antibody levels to NSP4 were modest (titers of < or = 200) in most of the infected and vaccinated children. A heterotypic NSP4 response was detected in 48% of the naturally rotavirus-infected children with a detectable response to NSP4. Following natural infection or RRV-TV vaccination, NSP4 was significantly less immunogenic than the VP6 protein when these responses were independently measured by ELISA. A significant (P < 0.05) proportion of children who did not develop diarrhea associated with rotavirus had antibodies to NSP4 in acute-phase serum, suggesting that serum antibodies against NSP4 might correlate with protection from rotavirus diarrhea. In addition, previous exposures to rotavirus did not affect the NSP4 seroconversion rate. PMID- 16210478 TI - Recombinant leishmania antigens for serodiagnosis of visceral leishmaniasis. AB - Serological tests with crude or recombinant Leishmania antigens are important tools for the diagnosis of leishmania infection. However, these tests are not markers of active visceral leishmaniasis (VL), since antibodies to these markers are often observed in individuals with subclinical L. chagasi infection and they do not fall shortly after therapy. In this study, levels of immunoglobulin G (IgG) against three recombinant Leishmania antigens (rH2A, KMP11, and the "Q" protein) were evaluated in sera from individuals with subclinical L. chagasi infection and in patients with VL pre- and posttherapy. The sensitivity of the serological test for diagnosis of VL was 100% with all three antigens. The titers of IgG fell significantly after therapy. While most of the individuals with subclinical L. chagasi infection had antibodies to rH2A and the "Q" protein, only 1 out of 15 individuals had antibodies to KMP11. These data indicate that KMP11 may be used to discriminate L. chagasi infection from active VL and may serve as a marker of response to therapy. PMID- 16210480 TI - Serologic cross-reactivity between Anaplasma marginale and Anaplasma phagocytophilum. AB - In the context of a serosurvey conducted on the Anaplasma marginale prevalence in Swiss cattle, we suspected that a serological cross-reactivity between A. marginale and A. phagocytophilum might exist. In the present study we demonstrate that cattle, sheep and horses experimentally infected with A. phagocytophilum not only develop antibodies to A. phagocytophilum (detected by immunofluorescent antibody assay) but also to A. marginale (detected by a competitive enzyme-linked immunosorbent assay). Conversely, calves experimentally infected with A. marginale also developed antibodies to A. phagocytophilum using the same serological tests. The identity of 63% determined in silico within a 209-amino acid sequence of major surface protein 5 of an isolate of A. marginale and one of A. phagocytophilum supported the observed immunological cross-reactivity. These observations have important consequences for the serotesting of both, A. marginale and A. phagocytophilum infection of several animal species. In view of these new findings, tests that have been considered specific for either infection must be interpreted carefully. PMID- 16210479 TI - Cryptococcus neoformans-reactive and total immunoglobulin profiles of human immunodeficiency virus-infected and uninfected Ugandans. AB - We determined total and Cryptococcus neoformans glucuronoxylomannan (GXM) reactive antibody repertoires of human immunodeficiency virus (HIV)-infected and HIV-uninfected Ugandans in a retrospective, case-control study of participants in a randomized controlled trial of pneumococcal vaccination. The study included 192 adults: 48 who subsequently developed cryptococcal meningitis (CM); (HIV+ CM+); 2 individuals who matched them in CD4+ T-cell level, stage of HIV disease, and age but did not develop CM (HIV+ CM-); and 48 HIV-uninfected individuals. Total serum immunoglobulin concentrations and titers of immunoglobulin M (IgM), IgG, and IgA to GXM, pneumococcal polysaccharides, and antibodies expressing certain V(H)3 idiotypes were determined with banked sera obtained before the development of cryptococcosis for HIV+ CM+ subjects. The results showed that HIV-infected subjects had significantly lower levels of IgM to GXM but higher levels of total immunoglobulin and IgG and IgA to GXM than those of HIV-uninfected subjects. HIV infected subjects with a history of pneumonia had higher levels, and those with a history of herpes zoster had lower levels of GXM-binding antibodies than subjects with no history of either disease. Minimal to no cross-reactivity was demonstrated between antibodies to GXM and polysaccharides in a pneumococcal vaccine. No significant differences between the antibody repertoires of HIV+ CM+ and HIV+ CM- subjects were identified, but among subjects without a history of pneumonia, there was a trend towards lower V(H)3-positive antibody levels among HIV+ CM+ than among HIV+ CM- subjects. Our findings demonstrate an association between previous infectious diseases and differences in the total and GXM reactive antibody repertoires of HIV-infected subjects and suggest the question of whether certain microbes modulate subsequent antibody responses to GXM deserves further study. PMID- 16210481 TI - Clonal diversity and turnover of Streptococcus mitis bv. 1 on shedding and nonshedding oral surfaces of human infants during the first year of life. AB - Streptococcus mitis bv. 1 is a pioneer colonizer of the human oral cavity. Studies of its population dynamics within parents and their infants and within neonates have shown extensive diversity within and between subjects. We examined the genetic diversity and clonal turnover of S. mitis bv. 1 isolated from the cheeks, tongue, and primary incisors of four infants from birth to 1 year of age. In addition, we compared the clonotypes of S. mitis bv. 1 isolated from their mothers' saliva collected in parallel to determine whether the mother was the origin of the clones colonizing her infant. Of 859 isolates obtained from the infants, 568 were unique clones. Each of the surfaces examined, whether shedding or nonshedding, displayed the same degree of diversity. Among the four infants it was rare to detect the same clone colonizing more than one surface at a given visit. There was little evidence for persistence of clones, but when clones were isolated on multiple visits they were not always found on the same surface. A similar degree of clonal diversity of S. mitis bv. 1 was observed in the mothers' saliva as in their infants' mouths. Clones common to both infant and mothers' saliva were found infrequently suggesting that this is not the origin of the infants' clones. It is unclear whether mucosal immunity exerts the environmental pressure driving the genetic diversity and clonal turnover of S. mitis bv. 1, which may be mechanisms employed by this bacterium to evade immune elimination. PMID- 16210482 TI - Potential of direct agglutination test based on promastigote and amastigote antigens for serodiagnosis of post-kala-azar dermal leishmaniasis. AB - Post-kala-azar dermal leishmaniasis (PKDL) is a dermal complication, a sequel to kala-azar. Diagnosis of PKDL presents a challenge due to the low parasite burden in the lesions. The direct agglutination test (DAT) based on promastigote and amastigote antigens of Leishmania donovani of indigenous isolates was developed to diagnose PKDL, and the results were compared with those of the rk39 strip test. The sensitivities of DAT for antileishmanial antibody detection, based on promastigote and amastigote antigens at a cutoff titer of 1:800 were 98.5% and 100%, respectively, with corresponding specificities of 96.5% and 100%. DAT could correctly detect 100% polymorphic cases and 95.4% macular PKDL cases. In comparison, the rk39 strip test was able to correctly diagnose 95.6% of polymorphic and 86.0% macular PKDL cases. DAT based on axenic amastigote antigen provided 100% sensitivity and specificity, making it particularly useful for macular PKDL cases, which are often missed by the rk39 strip test. Thus, DAT provides a simple, reliable, and inexpensive test for PKDL diagnosis with potential applicability in field conditions. PMID- 16210483 TI - Diagnosis of invasive pneumococcal disease among children in Kenya with enzyme linked immunosorbent assay for immunoglobulin G antibodies to pneumococcal surface adhesin A. AB - Diagnostic techniques for invasive pneumococcal disease (IPD) in children are insensitive and underestimate both the burden of disease and the cost effectiveness of pneumococcal conjugate vaccination (PCV). Consequently, there is little demand for the highly effective PCV outside the United States and Europe. In Kenya, diagnosis of pneumococcal pneumonia in adults was achieved with a sensitivity of 0.70 and a specificity of 0.98 using enzyme-linked immunosorbent assays (ELISAs) of paired plasma samples for immunoglobulin G (IgG) to pneumococcal surface adhesin A (PsaA). We aimed to validate the same technique in children. We assayed paired blood samples from 98 children with IPD, 95 age matched children with malaria/anemia, and 97 age-matched healthy controls by using an ELISA for anti-PsaA IgG. Sensitivity and specificity were determined in IPD patients and healthy controls. Specificity (0.97; 95% confidence interval [CI], 0.91 to 0.99) and sensitivity (0.42; 95% CI, 0.32 to 0.52) were optimized at a 2.7-fold rise in anti-PsaA antibody concentration. Sensitivity was improved to a maximum of 0.50 by restricting testing to children of <2 years old, by excluding IPD patients who were not sampled on the first day of presentation, and by incorporating high existing antibody concentrations in the analysis. Assay performance was independent of nasopharyngeal carriage of pneumococci at recruitment. This assay improves on existing diagnostic tools for IPD in children but would still leave over half of all cases undetected in epidemiological studies. Effective diagnosis of pneumococcal disease in children is urgently required but poorly served by existing technology. PMID- 16210484 TI - Generation of feline dendritic cells derived from peripheral blood monocytes for in vivo use. AB - Dendritic cells (DCs) are professional antigen-presenting cells that can prime T cells and polarize the cellular immune response. Because Th1-type immune responses have been connected to success in combating viral infection, a promising therapeutic application of DCs would be their differentiation in vitro and injection back into the host to boost an immune response in infected animals. This study was aimed both at developing a protocol to cultivate feline DCs in the absence of exogenous proteins for their use in vivo and at investigating what might be the most appropriate stimulus to induce their maturation in vitro and finding correlates of maturation. We generated DCs from peripheral blood monocytes in the presence of feline interleukin-4 and granulocyte-macrophage colony stimulating factor, and after 5 days their maturation was induced with either lipopolysaccharide, human recombinant tumor necrosis factor alpha, poly(I:C), or activated feline platelets. After 48 h, their CD14, CD1a, major histocompatibility complex class II, and B7.1 surface expression was analyzed in parallel with their ability to uptake antigen or prime a mixed leukocyte reaction. The results presented show that feline DCs cultured in autologous plasma differentiate and are able to mature in the presence of stimuli similar to the ones currently used for other species. The present work sets the grounds for future use of DCs obtained by the protocol described for in vivo vaccination and immunotherapy of feline immunodeficiency virus-infected cats. PMID- 16210485 TI - Feline coronavirus serotypes 1 and 2: seroprevalence and association with disease in Switzerland. AB - To determine the prevalence of antibodies to feline coronavirus (FCoV) serotypes 1 and 2 in Switzerland and their association with different disease manifestations, a serological study based on immunofluorescence tests was conducted with Swiss field cats using transmissible gastroenteritis virus (TGEV), FCoV type 1 and FCoV type 2 as antigens. A total of 639 serum samples collected in the context of different studies from naturally infected cats were tested. The current study revealed that, with an apparent prevalence of 83%, FCoV serotype 1 is the most prevalent serotype in Switzerland. FCoV type 1 viruses induced higher antibody titers than FCoV type 2, and were more frequently associated with clinical signs and/or feline infectious peritonitis. The antibody development in seven cats experimentally infected with FCoV type 1 revealed that, with progressing duration of infection, antibodies to FCoV type 1 significantly increased over those to FCoV type 2. There was a significant relationship between antibody titers against TGEV, FCoV 1, and FCoV 2 and TGEV antigen detected the highest proportion of seropositive cats. We conclude that a vaccine against FCoV should be based on FCoV type 1-related antigens and that for serodiagnosis of FCoV infection TGEV should be used to attain the highest diagnostic efficiency. When serology is used in addition to clinical signs, hematology, and clinical chemistry results as an aid to diagnose clinical FIP, TGEV shows a diagnostic efficiency equal to that of a FCoV antigen. PMID- 16210486 TI - Priming of immunological memory by pneumococcal conjugate vaccine in children unresponsive to 23-valent polysaccharide pneumococcal vaccine. AB - Pneumococcal polysaccharide vaccine (PPV) is of limited immunogenicity in infants and immunocompromised patients. Our prospective randomized controlled trial investigated whether priming with pneumococcal conjugate vaccine (PCV) induced specific immunological memory in previously nonresponders to PPV. Of a total of 33 children (2 to 18 years) with polysaccharide-specific immunodeficiency (PSI), group A (n = 16) received two doses of 7-valent PCV in a 4- to 6-week interval, and a booster dose of 23-valent PPV after one year. Group B (n = 17) received two doses of PPV in a 1-year interval exclusively. Specific antibody concentrations for serotypes 4, 5, 6B, 9V, 14, 18C, 19F, and 23F were determined (enzyme-linked immunosorbent assay) before and at 7 and 28 days after administration of the PPV booster and compared to an opsonophagocytosis assay. Of group A, 64 to 100% had antibody concentrations of > or = 1 microg/ml on day 28 after the booster versus 25 to 94% of group B. Group A had significantly higher antibody concentrations for all PCV-containing serotypes already on day 7, indicating early memory response. Antibody concentrations were in accordance with functional opsonic activity, although opsonic titers varied among individuals. Pneumococcal vaccination was well tolerated. The incidence of airway infections was reduced after priming with PCV (10/year for group A versus 15/year for group B). Following a PPV booster, even patients primarily not responding to PPV showed a rapid and more pronounced memory response after priming with PCV. PMID- 16210487 TI - Lassa fever virus peptides predicted by computational analysis induce epitope specific cytotoxic-T-lymphocyte responses in HLA-A2.1 transgenic mice. AB - Lassa fever is a hemorrhagic disease caused by Lassa fever virus (LV). Although the precise host defense mechanism(s) that affords protection against LV is not completely understood, cellular immunity mediated by cytotoxic T lymphocytes (CTLs) plays a pivotal role in controlling viral replication and LV infection. To date, there have been no reports mapping major histocompatibility complex (MHC) class I-binding CTL epitopes for LV. Using computer-assisted algorithms, we identified five HLA-A2.1-binding peptides of LV glycoprotein (GP) and two peptides from LV nucleoprotein (NP). Synthesized peptides were examined for their ability to bind to MHC class I molecules using a flow cytometric assay that measures peptide stabilization of class I. Three of the LV-GP peptides tested (LLGTFTWTL, SLYKGVYEL, and YLISIFLHL) stabilized HLA-A2. The LV-NP peptides tested failed to stabilize this HLA-A2. We then investigated the ability of the HLA-A2-binding LV-GP peptides to generate peptide-specific CTLs in HLA-A2.1 transgenic mice. Functional assays used to confirm CTL activation included gamma interferon enzyme-linked immunospot (ELISPOT) assays and intracellular cytokine staining of CD8+ T cells from peptide-primed mice. CTL assays were also performed to verify the cytolytic activity of peptide-pulsed target cells. Each of the LV GP peptides induced CTL responses in HLA-A2-transgenic mice. MHC class I tetramers prepared using one LV-GP peptide that showed the highest cytolytic index (LLGTFTWTL) confirmed that peptide-binding CD8+ T cells were present in pooled lymphocytes harvested from peptide-primed mice. These findings provide direct evidence for the existence of LV-derived GP epitopes that may be useful in the development of protective immunogens for this hemorrhagic virus. PMID- 16210488 TI - Evaluation of LIAISON Treponema Screen, a novel recombinant antigen-based chemiluminescence immunoassay for laboratory diagnosis of syphilis. AB - The purpose of this study was to evaluate the diagnostic performance of LIAISON Treponema Screen (DiaSorin, Saluggia, Italy), a new automated chemiluminescence immunoassay (CLIA), in comparison with that of rapid plasma reagin (RPR) and the following currently used treponemal tests: hemagglutination test (TPHA), immunoenzymatic assay (EIA), and Western blot (WB). First, a retrospective study was performed with a panel of 2,494 blood donor sera, a panel of 131 clinical and serologically characterized syphilitic sera, and 96 samples obtained from subjects with potentially interfering diseases or conditions. A prospective study was also performed by testing 1,800 unselected samples submitted to the Microbiology Laboratory of the St. Orsola Hospital in Bologna, Italy, for routine screening for syphilis. As expected, RPR was the least specific method, especially when potentially cross-reacting sera were tested. On the contrary, all of the treponemal tests proved to be very specific (99.9%) and they performed with the following sensitivities: 100% (WB), 99.2% (CLIA), 95.4% (EIA), and 94.7% (TPHA). PMID- 16210489 TI - Immunoglobulin A antibody responses in dengue patients: a useful marker for serodiagnosis of dengue virus infection. AB - We determined the usefulness of an immunoglobulin A (IgA) antibody-capture enzyme linked immunosorbent assay for serodiagnosis of dengue virus infections. The results indicate that the presence of IgA and IgM in serum samples assures recent primary dengue virus infection even with a single serum sample. PMID- 16210490 TI - Fluorescent multivalent opsonophagocytic assay for measurement of functional antibodies to Streptococcus pneumoniae. AB - We developed fluorescent mono- and multivalent opsonophagocytic assays (fOPA and fmOPA, respectively) specific for seven Streptococcus pneumoniae serotypes (4, 6B, 9V, 14, 18C, 19F, and 23F). Bacterial survival was quantitated with alamar blue, a fluorescent metabolic indicator. Both fOPA and fmOPA allow for determination of viability endpoints for up to seven serotypes with high levels of agreement to the reference method. The fmOPA eliminates colony counting, reduces serum volume, and produces results in 1 day. PMID- 16210491 TI - Immunogenicity of an intranasally administered modified live canine parvovirus type 2b vaccine in pups with maternally derived antibodies. AB - The ability of a modified live canine parvovirus type 2b vaccine to elicit active immunization in pups with maternally derived antibodies (MDA) by intranasal administration was evaluated. The vaccine induced seroconversion in 100% of pups with MDA titers of < or = 80 and in 51.6% of pups with titers between 160 and 320. PMID- 16210492 TI - Humoral and cellular immunity in children with Mycoplasma pneumoniae infection: a 1-year prospective study. AB - To determine whether children have persistent abnormalities in cellular and humoral immunity development after acute Mycoplasma pneumoniae infection, serum immunoglobulin G (IgG), IgA, IgM, and IgE levels and lymphocyte phenotypes were determined. There were no changes in the levels of IgG, IgM, IgA, or CD4+ or CD19+ lymphocytes that were measured in M. pneumoniae-positive patients after 3 months or after 12 months, but there were increases in these in M. pneumoniae negative patients. Serum IgE increased in M. pneumoniae-positive patients. We have shown alterations in immunity development after M. pneumoniae infection. PMID- 16210493 TI - Immunogenicity of meningococcal ACYW135 polysaccharide vaccine in Saudi children 5 to 9 years of age. AB - Meningococcal tetravalent polysaccharide vaccines were observed to be immunogenic in Saudi children 5 to 9 years of age, with >90% having serum bactericidal antibody titers of > or = 8 for serogroups A, Y, and W135; for serogroup C, 77% were putatively protected after vaccination. PMID- 16210494 TI - Critical investigation of the CD14 promoter polymorphism: lack of a role for in vitro cytokine response and membrane CD14 expression. AB - Blood of volunteers, genotyped for the CD14 C(-159)-->T polymorphism, showed no difference in cytokine release when stimulated with nine CD14-dependent immune stimuli. An analysis of the published data on the proposed association of CD14 genotype with membrane CD14 density revealed no significant correlation, questioning a functional impact of the CD14 polymorphism. PMID- 16210495 TI - Increased levels of macrophage migration inhibitory factor in sera of patients with Escherichia coli O157:H7-induced enterocolitis. PMID- 16210496 TI - 37-Kilodalton/83-kilodalton RNase L isoform ratio in peripheral blood mononuclear cells: analytical performance and relevance for chronic fatigue syndrome. PMID- 16210497 TI - Stem-cell homeostasis and growth dynamics can be uncoupled in the Arabidopsis shoot apex. AB - The shoot apical meristem (SAM) is a collection of stem cells that resides at the tip of each shoot and provides the cells of the shoot. It is divided into functional regions. The central zone (CZ) at the tip of the meristem is the domain of expression of the CLAVATA3 (CLV3) gene, encoding a putative ligand for a transmembrane receptor kinase, CLAVATA1, active in cells of the rib meristem (RM), located just below the CZ. We show here that CLV3 restricts its own domain of expression (the CZ) by preventing differentiation of peripheral zone cells (PZ), which surround the CZ, into CZ cells and restricts overall SAM size by a separate, long-range effect on cell division rate. PMID- 16210498 TI - Ordered liquid aluminum at the interface with sapphire. AB - Understanding the nature of solid-liquid interfaces is important for many processes of technological interest, such as solidification, liquid-phase epitaxial growth, wetting, liquid-phase joining, crystal growth, and lubrication. Recent studies have reported on indirect evidence of density fluctuations at solid-liquid interfaces on the basis of x-ray scattering methods that have been complemented by atomistic simulations. We provide evidence for ordering of liquid atoms adjacent to an interface with a crystal, based on real-time high temperature observations of alumina-aluminum solid-liquid interfaces at the atomic-length scale. In addition, crystal growth of alumina into liquid aluminum, facilitated by interfacial transport of oxygen from the microscope column, was observed in situ with the use of high-resolution transmission electron microscopy. PMID- 16210499 TI - The radiative signature of upper tropospheric moistening. AB - Climate models predict that the concentration of water vapor in the upper troposphere could double by the end of the century as a result of increases in greenhouse gases. Such moistening plays a key role in amplifying the rate at which the climate warms in response to anthropogenic activities, but has been difficult to detect because of deficiencies in conventional observing systems. We use satellite measurements to highlight a distinct radiative signature of upper tropospheric moistening over the period 1982 to 2004. The observed moistening is accurately captured by climate model simulations and lends further credence to model projections of future global warming. PMID- 16210500 TI - 1918 flu and responsible science. PMID- 16210502 TI - U.S. biomedical policy. Acting FDA head drops NCI post. PMID- 16210501 TI - Virology. Resurrected influenza virus yields secrets of deadly 1918 pandemic. PMID- 16210503 TI - Neuroscience. Cancer drugs may help injured nerve cells regrow their axons. PMID- 16210505 TI - Conservation policy. House revises Endangered Species Act. PMID- 16210504 TI - Ecology. Satellite tracking catches sharks on the move. PMID- 16210506 TI - France. Reform law fails to impress researchers. PMID- 16210507 TI - Physiology or medicine. Triumph of the ulcer-bug theory. PMID- 16210508 TI - Scientific publishing. Withdrawn parasite paper stirs criticism of cell. PMID- 16210509 TI - Physics. Quantum optics shines in the photon's centenary. PMID- 16210510 TI - Endangered species. Ban on Beluga caviar points to sturgeon's worldwide decline. PMID- 16210511 TI - Astrophysics. Short gamma ray bursts: mystery solved. PMID- 16210512 TI - Drug regulation. Plan B: A collision of science and politics. PMID- 16210513 TI - Conservation biology. Premier Latin American institute loses grants, ponders future. PMID- 16210514 TI - Russian science. Academy agrees to post-soviet crash diet. PMID- 16210515 TI - Neurodegeneration. Huntington's research points to possible new therapies. PMID- 16210516 TI - Retraction. Hairpin RNAs and retrotransposon LTRs effect RNAi and chromatin-based gene silencing. PMID- 16210517 TI - Benefits and risks in malaria control. PMID- 16210518 TI - Estrogen receptors and cell signaling. PMID- 16210519 TI - What should we call Pluto? PMID- 16210520 TI - Global voices of science: Mangroves, fishponds, and the quest for sustainability. AB - Aquaculture, the farming of shrimp and other useful aquatic and marine plants and animals in artificially confined and tended ponds, pens, and cages, ranks as a phenomenal success story in global food production. In 1975, aquaculture contributed 8% to the overall yield of the world's fish harvest; now it provides more than one-third of the yield. Total aquaculture production in 2003 was 54.8 million metric tons valued at 67.3 billion in U.S. dollars. More than 90% of this output comes from Asia, where aquaculture has its origins and where this month's essay author has lived and worked all of her life. In her essay, Jurgenne H. Primavera, senior scientist of the Aquaculture Department of the Southeast Asian Fisheries Development Center based in Iloilo, Central Philippines, traces the recent history of aquaculture and the socioeconomic and environmental challenges that its rapid growth has wrought, especially for the mangrove ecosystems in which much of brackishwater pond aquaculture occurs. With an eye on all stakeholders, Primavera lays out how aquaculture is now falling short of the goal of sustainability and what steps might be taken to move the industry in that direction. PMID- 16210521 TI - Plant biology. Enhanced: growth by auxin: when a weed needs acid. AB - In his Perspective, Grebe discusses how a plant proton pump residing in intracellular compartments, rather than in the plasma membrane of the cell surface, regulates growth and development. The pump modulates the expression at the plasma membrane of both a transporter for the hormone auxin and another proton pump. These findings open new views on how plants regulate cell wall acidity and hormone transport during development. PMID- 16210522 TI - Applied physics. Subsurface imaging with scanning ultrasound holography. AB - Every new microscopic imaging technique reveals hidden features but also new challenges. To capture information about substructure features, especially defects and voids, in the next generation of integrated circuits, higher resolution methods of surface imaging will be required. In his Perspective, Diebold discusses results reported in the same issue by Shekhawat and Dravid in which an acoustic scanning holographic imaging technique has been extended to unprecedented spatial resolution. The method has also been used on biological cells, and the hope is that it can be developed further to obtain detailed information about the depth and elastic properties of buried features. PMID- 16210523 TI - Psychology. The nature of personality: genes, culture, and national character. AB - Personality traits have a strong genetic foundation, are highly stable over time, and predict important societal outcomes, including health and occupational success. In his Perspective, Robins discusses Terracciano et al.'s finding that cultures differ somewhat in aggregate personality levels but those differences are not accurately reflected in stereotypes about national character. Robins discusses reasons why national stereotypes are inaccurate, as well as broader issues concerning individual and cultural sources of variation in personality. PMID- 16210524 TI - Chemistry. Inventing the nanomolecular wheel. AB - The wheel is considered the canonical invention leading to all manner of mechanical devices. Rotating machinery is also being investigated at the nanoscale as researchers attempt to create molecular analogs of wheels and motors. But will these devices be merely imitations of macroscopic machines? In his Perspective, Siegel discusses results reported in the same issue by Fletcher et al. in which a four-stroke chemical nanoengine is reported. By a series of bond-breaking and bond-forming steps, a molecular structure is caused to undergo directed rotation. The development of structures of this kind, which are not merely miniaturized macromachines, may lead to other devices for executing controlled oscillatory motion. PMID- 16210526 TI - Connected to death: the (unexpurgated) mitochondrial pathway of apoptosis. AB - The mitochondrial pathway of apoptosis in vertebrates is dependent on the process of mitochondrial outer membrane permeabilization (MOMP), which leads to the release of proteins from the mitochondrial intermembrane space into the cytosol. "Upstairs" of this event are the Bcl-2 family proteins that regulate and mediate MOMP; "downstairs" is the activation of caspases that orchestrate the dismantling of the cell. In the Connections Map database at Science's Signal Transduction Knowledge Environment (STKE), the pathways that define the mitochondrial pathway of apotosis are illustrated, with the bulk of control occurring "upstairs" of MOMP. PMID- 16210527 TI - Transforming growth factor-beta signaling in stem cells and cancer. AB - Transforming growth factor-beta (TGF-beta) and TGF-beta-related proteins, such as the bone morphogenetic proteins, have emerged as key regulators of stem cell renewal and differentiation. These proteins have disparate roles in regulating the biology of embryonic stem cells and tumor suppression, and they help define the selection of cell fate and the progression of differentiation along a lineage. Here we illustrate their roles in embryonic stem cells and in the differentiation of neural, hematopoietic, mesenchymal, and gastrointestinal epithelial stem cells. PMID- 16210528 TI - G proteins Go green: a plant G protein signaling FAQ sheet. AB - Plants, like animals, use signal transduction pathways based on heterotrimeric guanine nucleotide-binding proteins (G proteins) to regulate many aspects of development and cell signaling. Some components of G protein signaling are highly conserved between plants and animals and some are not. This Viewpoint compares key aspects of G protein signal transduction in plants and animals and describes the current knowledge of this system in plants, the questions that still await exploration, and the value of research on plant G proteins to scientists who do not study plants. Pathways in Science's Signal Transduction Knowledge Environment Connections Maps database provide details about the emerging roles of G proteins in several cellular processes of plants. PMID- 16210529 TI - Bottom-feeding plesiosaurs. AB - Elasmosaurid plesiosaurs were an important part of Cretaceous marine reptile communities and are generally considered to have been predators of small, agile, free-swimming fish and cephalopods. Two elasmosaurid specimens from Aptian and Albian deposits in Queensland, Australia, include fossilized gut contents dominated by benthic invertebrates: bivalves, gastropods, and crustaceans. Both specimens also contained large numbers of gastroliths (stomach stones). These finds point to a wider niche than has previously been supposed for these seemingly specialized predators and may also influence long-running controversy over the question of gastrolith function in plesiosaurs. PMID- 16210531 TI - A reversible, unidirectional molecular rotary motor driven by chemical energy. AB - With the long-term goal of producing nanometer-scale machines, we describe here the unidirectional rotary motion of a synthetic molecular structure fueled by chemical conversions. The basis of the rotation is the movement of a phenyl rotor relative to a naphthyl stator about a single bond axle. The sense of rotation is governed by the choice of chemical reagents that power the motor through four chemically distinct stations. Within the stations, the rotor is held in place by structural features that limit the extent of the rotor's Brownian motion relative to the stator. PMID- 16210530 TI - Characterization of the reconstructed 1918 Spanish influenza pandemic virus. AB - The pandemic influenza virus of 1918-1919 killed an estimated 20 to 50 million people worldwide. With the recent availability of the complete 1918 influenza virus coding sequence, we used reverse genetics to generate an influenza virus bearing all eight gene segments of the pandemic virus to study the properties associated with its extraordinary virulence. In stark contrast to contemporary human influenza H1N1 viruses, the 1918 pandemic virus had the ability to replicate in the absence of trypsin, caused death in mice and embryonated chicken eggs, and displayed a high-growth phenotype in human bronchial epithelial cells. Moreover, the coordinated expression of the 1918 virus genes most certainly confers the unique high-virulence phenotype observed with this pandemic virus. PMID- 16210532 TI - Sequential proton transfer through water bridges in acid-base reactions. AB - The proton transfer mechanism between aqueous Bronsted acids and bases, forming an encounter pair, has been studied in real time with ultrafast infrared spectroscopy. The transient intermediacy of a hydrated proton, formed by ultrafast dissociation from an optically triggered photoacid proton donor ROH, is implicated by the appearance of an infrared absorption marker band before protonation of the base, B-. Thus, proton exchange between an acid and a base in aqueous solution is shown to proceed by a sequential, von Grotthuss-type, proton hopping mechanism through water bridges. The spectra suggest a hydronium cation H3O+ structure for the intermediate, stabilized in the Eigen configuration in the ionic complex RO-...H3O+...B-. PMID- 16210533 TI - PbSe nanocrystal solids for n- and p-channel thin film field-effect transistors. AB - Initially poorly conducting PbSe nanocrystal solids (quantum dot arrays or superlattices) can be chemically "activated" to fabricate n- and p-channel field effect transistors with electron and hole mobilities of 0.9 and 0.2 square centimeters per volt-second, respectively; with current modulations of about 10(3) to 10(4); and with current density approaching 3 x 10(4) amperes per square centimeter. Chemical treatments engineer the interparticle spacing, electronic coupling, and doping while passivating electronic traps. These nanocrystal field effect transistors allow reversible switching between n- and p-transport, providing options for complementary metal oxide semiconductor circuits and enabling a range of low-cost, large-area electronic, optoelectronic, thermoelectric, and sensing applications. PMID- 16210534 TI - Nanoscale imaging of buried structures via scanning near-field ultrasound holography. AB - A nondestructive imaging method, scanning near-field ultrasound holography (SNFUH), has been developed that provides depth information as well as spatial resolution at the 10- to 100-nanometer scale. In SNFUH, the phase and amplitude of the scattered specimen ultrasound wave, reflected in perturbation to the surface acoustic standing wave, are mapped with a scanning probe microscopy platform to provide nanoscale-resolution images of the internal substructure of diverse materials. We have used SNFUH to image buried nanostructures, to perform subsurface metrology in microelectronic structures, and to image malaria parasites in red blood cells. PMID- 16210535 TI - A 5-micron-bright spot on Titan: evidence for surface diversity. AB - Observations from the Cassini Visual and Infrared Mapping Spectrometer show an anomalously bright spot on Titan located at 80 degrees W and 20 degrees S. This area is bright in reflected light at all observed wavelengths, but is most noticeable at 5 microns. The spot is associated with a surface albedo feature identified in images taken by the Cassini Imaging Science Subsystem. We discuss various hypotheses about the source of the spot, reaching the conclusion that the spot is probably due to variation in surface composition, perhaps associated with recent geophysical phenomena. PMID- 16210536 TI - National character does not reflect mean personality trait levels in 49 cultures. AB - Most people hold beliefs about personality characteristics typical of members of their own and others' cultures. These perceptions of national character may be generalizations from personal experience, stereotypes with a "kernel of truth," or inaccurate stereotypes. We obtained national character ratings of 3989 people from 49 cultures and compared them with the average personality scores of culture members assessed by observer ratings and self-reports. National character ratings were reliable but did not converge with assessed traits. Perceptions of national character thus appear to be unfounded stereotypes that may serve the function of maintaining a national identity. PMID- 16210537 TI - Transoceanic migration, spatial dynamics, and population linkages of white sharks. AB - The large-scale spatial dynamics and population structure of marine top predators are poorly known. We present electronic tag and photographic identification data showing a complex suite of behavioral patterns in white sharks. These include coastal return migrations and the fastest known transoceanic return migration among swimming fauna, which provide direct evidence of a link between widely separated populations in South Africa and Australia. Transoceanic return migration involved a return to the original capture location, dives to depths of 980 meters, and the tolerance of water temperatures as low as 3.4 degrees C. These findings contradict previous ideas that female white sharks do not make transoceanic migrations, and they suggest natal homing behavior. PMID- 16210538 TI - Satellite tagging and cardiac physiology reveal niche expansion in salmon sharks. AB - Shark populations are declining globally, yet the movements and habitats of most species are unknown. We used a satellite tag attached to the dorsal fin to track salmon sharks (Lamna ditropis) for up to 3.2 years. Here we show that salmon sharks have a subarctic-to-subtropical niche, ranging from 2 degrees to 24 degrees C, and they spend winter periods in waters as cold as 2 degrees to 8 degrees C. Functional assays and protein gels reveal that the expression of excitation-contraction coupling proteins is enhanced in salmon shark hearts, which may underlie the shark's ability to maintain heart function at cold temperatures and their niche expansion into subarctic seas. PMID- 16210539 TI - EGFR activation mediates inhibition of axon regeneration by myelin and chondroitin sulfate proteoglycans. AB - Inhibitory molecules associated with myelin and the glial scar limit axon regeneration in the adult central nervous system (CNS), but the underlying signaling mechanisms of regeneration inhibition are not fully understood. Here, we show that suppressing the kinase function of the epidermal growth factor receptor (EGFR) blocks the activities of both myelin inhibitors and chondroitin sulfate proteoglycans in inhibiting neurite outgrowth. In addition, regeneration inhibitors trigger the phosphorylation of EGFR in a calcium-dependent manner. Local administration of EGFR inhibitors promotes significant regeneration of injured optic nerve fibers, pointing to a promising therapeutic avenue for enhancing axon regeneration after CNS injury. PMID- 16210540 TI - Retinoic acid signaling affects cortical synchrony during sleep. AB - Delta oscillations, characteristic of the electroencephalogram (EEG) of slow wave sleep, estimate sleep depth and need and are thought to be closely linked to the recovery function of sleep. The cellular mechanisms underlying the generation of delta waves at the cortical and thalamic levels are well documented, but the molecular regulatory mechanisms remain elusive. Here we demonstrate in the mouse that the gene encoding the retinoic acid receptor beta determines the contribution of delta oscillations to the sleep EEG. Thus, retinoic acid signaling, which is involved in the patterning of the brain and dopaminergic pathways, regulates cortical synchrony in the adult. PMID- 16210541 TI - Astrocytic purinergic signaling coordinates synaptic networks. AB - To investigate the role of astrocytes in regulating synaptic transmission, we generated inducible transgenic mice that express a dominant-negative SNARE domain selectively in astrocytes to block the release of transmitters from these glial cells. By releasing adenosine triphosphate, which accumulates as adenosine, astrocytes tonically suppressed synaptic transmission, thereby enhancing the dynamic range for long-term potentiation and mediated activity-dependent, heterosynaptic depression. These results indicate that astrocytes are intricately linked in the regulation of synaptic strength and plasticity and provide a pathway for synaptic cross-talk. PMID- 16210542 TI - Failure to detect mismatches between intention and outcome in a simple decision task. AB - A fundamental assumption of theories of decision-making is that we detect mismatches between intention and outcome, adjust our behavior in the face of error, and adapt to changing circumstances. Is this always the case? We investigated the relation between intention, choice, and introspection. Participants made choices between presented face pairs on the basis of attractiveness, while we covertly manipulated the relationship between choice and outcome that they experienced. Participants failed to notice conspicuous mismatches between their intended choice and the outcome they were presented with, while nevertheless offering introspectively derived reasons for why they chose the way they did. We call this effect choice blindness. PMID- 16210543 TI - Sexual selection can resolve sex-linked sexual antagonism. AB - Sexual selection is a potent evolutionary force. However, very few models have considered the evolution of female preferences for traits expressed in both sexes. Here we explore how female preferences coevolve with sexually antagonistic traits, which involve alleles that are beneficial to one sex but harmful to the other. We show that with a sexually antagonistic trait on the X chromosome (males XY, females XX), females evolve to prefer mates carrying alleles beneficial to daughters. In contrast, with a Z-linked trait (males ZZ, females ZW), females more often evolve mating preferences for mates carrying alleles beneficial to sons (that is, flashy displays). PMID- 16210544 TI - Arabidopsis H+-PPase AVP1 regulates auxin-mediated organ development. AB - The transport of auxin controls developmental events in plants. Here, we report that in addition to maintaining vacuolar pH, the H+-pyrophosphatase, AVP1, controls auxin transport and consequently auxin-dependent development. AVP1 overexpression results in increased cell division at the onset of organ formation, hyperplasia, and increased auxin transport. In contrast, avp1-1 null mutants have severely disrupted root and shoot development and reduced auxin transport. Changes in the expression of AVP1 affect the distribution and abundance of the P-adenosine triphosphatase and Pinformed 1 auxin efflux facilitator, two proteins implicated in auxin distribution. Thus, AVP1 facilitates the auxin fluxes that regulate organogenesis. PMID- 16210545 TI - Haplotype effect of the matrix metalloproteinase-1 gene on risk of myocardial infarction. AB - Myocardial infarction (MI) is commonly caused by atherosclerotic plaque rupture following excessive degradation of collagen fibers in the atherosclerotic lesion. We investigated whether interindividual variability in risk of MI was related to polymorphisms in the gene encoding matrix metalloproteinase (MMP)-1, a key fibrillar collagen-degrading enzyme. Several single nucleotide polymorphisms in the MMP1 gene promoter were identified following sequencing DNA samples from 30 individuals. An analysis of the polymorphisms in a cohort of British whites with coronary atherosclerosis, including 639 patients with MI and 538 non-MI subjects, revealed a haplotype effect of the -519A>G and -340T>C polymorphisms on risk of MI, with the A(-519)-C(-340) and G(-519)-T(-340) haplotypes being protective (odds ratio=0.70 [0.57 to 0.86]; P=0.0007), whereas the G(-519)-C(-340) haplotype increased MI risk (odds ratio=1.94 [1.15 to 3.28]; P=0.013). This finding was replicated in a subsequent analysis of 387 Swedish MI patients and 387 healthy controls (odds ratio=0.70 [0.55 to 0.89], P=0.003, for A(-519)-C(-340) and G( 519)-T(-340); odds ratio=1.54 [0.97 to 2.46], P=0.07, for G(-519)-C(-340)). In vitro assays showed that compared with the A(-519)-T(-340) haplotype, the A(-519) C(-340) and G(-519)-T(-340) haplotypes had lower promoter activity, whereas the G(-519)-C(-340) haplotype had greater promoter strength, in driving gene expression in human macrophages. Haplotype-specific differences in MMP1 mRNA level in atherosclerotic tissues were also detected. The data indicate that MMP1 gene variation is a genetic factor contributing to interindividual differences in MI risk. PMID- 16210546 TI - NADPH oxidase activity and function are profoundly greater in cerebral versus systemic arteries. AB - Recent studies suggest that the superoxide generating enzyme NADPH oxidase may play a functional role in regulating cerebral vascular tone. We tested whether the activity, function, and expression of NADPH oxidase differs between rat cerebral and systemic arteries. Superoxide production by basilar (BA), middle cerebral (MCA), carotid (CA), renal (RA), and mesenteric (MA) arteries and aorta (AO) was measured using lucigenin-enhanced chemiluminescence. Superoxide production from NADPH oxidase was localized and semiquantified using dihydroethidium. Vascular functional responses were assessed in a myograph or organ bath. Vascular Nox4 protein expression was measured using Western blotting. Superoxide production (basal or in response to NADPH or angiotensin II) in the intracranial arteries, BA, and MCA was 10- to 100-fold greater than in AO, CA, RA, or MA. Similar results were found using either intact vessels or arterial homogenates, and were associated with 10-fold greater expression of Nox4 in the BA versus AO, CA, and MA. Superoxide production was attenuated by the NADPH oxidase inhibitors, diphenyleneiodonium, apocynin, and gp91ds-tat. NADPH and H2O2 were strong relaxing stimuli in the BA, where the H2O2 scavenger catalase, as well as apocynin, attenuated these relaxations and also augmented contractions to angiotensin II. NADPH oxidase activity is markedly higher in intracranial versus systemic arteries, in association with higher Nox4 expression. In cerebral arteries, endogenous H2O2 derived from NADPH oxidase activation appears to cause relaxation and is able to offset angiotensin II-induced constriction. These data are consistent with the concept that NADPH oxidase-derived reactive oxygen species modulate cerebral vascular tone under physiological conditions. PMID- 16210547 TI - Ca2+ influx-induced sarcoplasmic reticulum Ca2+ overload causes mitochondrial dependent apoptosis in ventricular myocytes. AB - Increases in Ca2+ influx through the L-type Ca2+ channel (LTCC, Cav1.2) augment sarcoplasmic reticulum (SR) Ca2+ loading and the amplitude of the cytosolic Ca2+ transient to enhance cardiac myocyte contractility. Our hypothesis is that persistent increases in Ca2+ influx through the LTCC cause apoptosis if the excessive influx results in SR Ca2+ overload. Feline ventricular myocytes (VMs) in primary culture were infected with either an adenovirus (Ad) containing a rat Cav1.2 beta2a subunit-green fluorescent protein (GFP) fusion gene (Adbeta2a) to increase Ca2+ influx or with AdGFP as a control. Significantly fewer beta2a-VMs (21.4+/-5.6%) than GFP-VMs (99.6+/-1.7%) were viable at 96 hours. A fraction of beta2a-VMs (20.8+/-1.8%) contracted spontaneously (SC-beta2a-VMs), and viability was significantly correlated with the percentage of SC-beta2a-VMs. Higher percentages of apoptotic nuclei, DNA laddering, and cytochrome C release were detected in beta2a-VMs. This apoptosis was prevented with pancaspase or caspase-3 or caspase-9 inhibitors. L-type calcium current (I(Ca-L)) density was greater in beta2a-VMs (23.4+/-2.8 pA/pF) than in GFP-VMs (7.6+/-1.6 pA/pF). SC-beta2a-VMs had higher diastolic intracellular Ca2+ (Indo-1 ratio: 1.1+/-0.1 versus 0.7+/ 0.03, P<0.05) and systolic Ca2+ transients (1.89+/-0.27 versus 0.80+/-0.08) than GFP-VMs. Inhibitors of Ca2+ influx, SR Ca2+ uptake and release, mitochondrial Ca2+ uptake, mitochondrial permeation transition pore, calpain, and Bcl-2 associated X protein protected beta2a-VMs from apoptosis. These results show that persistent increases in Ca2+ influx through the I(Ca-L) enhance contractility but lead to apoptosis through a mitochondrial death pathway if SR Ca2+ overload is induced. PMID- 16210548 TI - Intrasarcoplasmic amyloidosis impairs proteolytic function of proteasomes in cardiomyocytes by compromising substrate uptake. AB - The presence of increased ubiquitinated proteins and amyloid oligomers in failing human hearts strikingly resembles the characteristic pathology in the brain of many neurodegenerative diseases. The ubiquitin-proteasome system (UPS) is responsible for degradation of most cellular proteins and plays essential roles in virtually all cellular processes. UPS impairment by aberrant protein aggregation was previously shown in cell culture but remains to be demonstrated in intact animals. Mechanisms underlying the impairment are poorly understood. We report here that UPS proteolytic function is severely impaired in the heart of a mouse model of intrasarcoplasmic amyloidosis caused by cardiac-restricted expression of a human desmin-related myopathy-linked missense mutation of alphaB crystallin (CryAB(R120G)). The UPS impairment was detected before cardiac hypertrophy, and failure became discernible, suggesting that defective protein turnover likely contributes to cardiac remodeling and failure in this model. Further analyses reveal that the impairment is likely attributable to insufficient delivery of substrate proteins into the 20S proteasomes, and depletion of key components of the 19S subcomplex may be responsible. The derangement is likely caused by aberrant protein aggregation rather than loss of function of the CryAB gene because UPS malfunction was not evident in CryAB-null hearts and inhibition of aberrant protein aggregation by Congo red or a heat shock protein significantly attenuated CryAB(R120G)-induced UPS malfunction in cultured cardiomyocytes. Because of the central role of the UPS in cell regulation and the high intrasarcoplasmic amyloidosis prevalence in failing human hearts, our data suggest a novel pathogenic process in cardiac disorders with abnormal protein aggregation. PMID- 16210549 TI - Calreticulin destabilizes glucose transporter-1 mRNA in vascular endothelial and smooth muscle cells under high-glucose conditions. AB - Substrate autoregulation of glucose transporter-1 (GLUT-1) mRNA and protein expression provides vascular endothelial and smooth muscle cells a sensitive mechanism to adapt their rate of glucose transport in response to changing glycemic conditions. Hyperglycemia-induced downregulation of glucose transport is particularly important in protecting these cells against an excessive influx of glucose and consequently increased intracellular protein glycation and generation of free radicals; both are detrimental in the development of vascular disease in diabetes. We aimed to investigate the molecular mechanism of high glucose-induced downregulation of GLUT-1 mRNA expression in primary bovine aortic vascular endothelial (VEC) and smooth muscle (VSMC) cell cultures. Using RNA mobility shift, UV cross-linking, and in vitro degradation assays, followed by mass spectrometric analysis, we identified calreticulin as a specific destabilizing trans-acting factor that binds to a 10-nucleotide cis-acting element (CAE(2181 2190)) in the 3'-untranslated region of GLUT-1 mRNA. Pure calreticulin accelerated the rate of GLUT-1 mRNA-probe degradation in vitro, whereas overexpression of calreticulin in vascular cells decreased significantly the total cell content of GLUT-1 mRNA and protein. The expression of calreticulin was augmented in vascular cells exposed to high glucose in comparison with low glucose conditions. Similarly, increased expression of calreticulin was observed in aortae of diabetic Psammomys obesus in comparison with normoglycemic controls. These data suggest that CAE(2181-2190)-calreticulin complex, which is formed in VSMC and VEC exposed to hyperglycemic conditions, renders GLUT-1 mRNA susceptible to degradation. This interaction underlies the process of downregulation of glucose transport in vascular cells under high-glucose conditions. PMID- 16210550 TI - Mobilization of bone marrow-derived cells enhances the angiogenic response to hypoxia without transdifferentiation into endothelial cells. AB - Bone marrow-derived cells (BMCs) have been implicated as a modifiers of vascular growth either directly by transdifferentiation into endothelial cells (ECs) or indirectly through growth factor release. To examine these possibilities under physiological conditions, we developed a model of hypoxia-mediated angiogenesis in the mouse spinotrapezius muscle. This allows whole-mount analysis; therefore, the morphology and location of BMCs within the vascular network may be observed along with differentiation markers. We exposed bone marrow transplant chimeric mice to hypoxia and treated a subset with granulocyte macrophage colony stimulating factor. Exposure to hypoxia caused an 13% increase in capillary density relative to control. Hypoxia did not increase the overall number of muscle-resident BMCs, but did increase the number of rounded BMCs by 25%. There was no discernable BMC contribution to the endothelium, although some BMCs assumed a pericyte morphology around capillaries. Granulocyte macrophage colony stimulating factor treatment further increased the number of round BMCs within the muscle and caused a 23% increase in angiogenesis. The results of this study suggest a potentially beneficial action of BMCs during hypoxia through paracrine release of growth factors but not transdifferentiation into ECs. PMID- 16210551 TI - Thrombolysis beyond the guidelines: two treatments in one subject within 90 hours based on a modified magnetic resonance imaging brain clock concept. AB - BACKGROUND AND PURPOSE: We report the first case of 2 intravenous thrombolysis treatments within 90 hours in a patient with early recurrent stroke. SUMMARY OF REVIEW: A 50-year-old man had improved significantly after intravenous thrombolysis for acute stroke. On the fourth day, he deteriorated dramatically because of recurrent stroke. Evidence of vessel reocclusion and profound perfusion/diffusion mismatch constituted the rationale for a second thrombolysis treatment, which resulted in vessel recanalization and significant neurologic improvement. CONCLUSIONS: The pathophysiological information obtained by multimodal magnetic resonance imaging may suit as a brain clock when repeat thrombolysis treatment is considered for early recurrent stroke. PMID- 16210552 TI - Poor outcomes in patients who do not receive intravenous tissue plasminogen activator because of mild or improving ischemic stroke. AB - BACKGROUND AND PURPOSE: Some patients with mild or improving ischemic stroke symptoms do not receive intravenous tissue plasminogen activator (tPA) because they look "too good to treat" (TGT); however, some have poor outcomes. METHODS: We retrospectively analyzed data from a prospective single-center study between 2002 and 2004. TGT patients were those arriving within 3 hours of symptom onset and not treated with intravenous tPA solely because of mild or improving symptoms. RESULTS: Of 128 patients presenting within 3 hours, 41 (34%) were not given tPA because of mild or improving stroke. Of the TGT patients, 11 of 41 (27%) died or were not discharged home because of neurological worsening (n=6) or persistent "mild" neurological deficit (n=5). No single variable at presentation was associated with death or lack of home discharge. There were 10 of 41 TGT patients (24%) who had > or =4-point improvement in National Institutes of Health Stroke Scale score before tPA decision; these patients were more likely to have subsequent neurological worsening (relative risk, 4.1, 95% CI, 1.1 to 15.4; P=0.05). CONCLUSIONS: A substantial minority of patients deemed too good for intravenous tPA were unable to be discharged home. A re-evaluation of the stroke severity criteria for tPA eligibility may be indicated. PMID- 16210553 TI - Supporting family caregivers in stroke care: a review of the evidence for problem solving. AB - BACKGROUND AND PURPOSE: Teaching effective problem-solving skills to family caregivers of patients with chronic disease has been shown to be useful for promoting physical and psychosocial well-being. However, the use and effectiveness of problem solving for supporting caregivers in stroke care has not been reviewed. This article aims to identify and review studies that have examined the effectiveness of teaching problem solving skills to caregivers in stroke care, highlight gaps in the evidence base, and recommend avenues for additional research. METHODS: A structured review of literature identified from nursing, medicine, and psychology databases from 1970 to 2004 was conducted. Eleven articles reporting the development or evaluation of effective problem solving interventions for caregivers of patients with stroke were critically appraised using recognized quality criteria. RESULTS: The results of this review show that the strength of evidence for problem-solving interventions for caregivers of stroke patients is limited. Because some studies used small samples and varied methods and interventions, making a comparison was difficult. Caregivers' problem-solving abilities were rarely measured, and the theoretical concepts and framework underpinning most studies were unclear. CONCLUSIONS: Evidence from the review suggests a need to additionally study the link between theoretical concepts of effective problem solving and outcomes using standardized measures and to examine also the processes involved in implementing the intervention using multimethod designs, including both quantitative and qualitative approaches. PMID- 16210554 TI - Outcome and severe hemorrhagic complications of intravenous thrombolysis with tissue plasminogen activator in very old (> or =80 years) stroke patients. AB - BACKGROUND AND PURPOSE: Information on safety and efficacy of intravenous thrombolysis with tissue plasminogen activator (tPA) (IV-tPA) in very old acute ischemic stroke (AIS) patients is scarce. We studied outcome and severe hemorrhagic complications in patients aged 80 and older. METHODS: We analyzed data of AIS patients, treated with IV-tPA, in 3 German stroke centers. Neurologic deficit on admission was assessed using the National Institutes of Health Stroke Scale (NIHSS). Outcome was assessed after 90 days using the Modified Rankin Scale (MRS), and favorable outcome was defined as a MRS score of 0 to 1. Severe intracerebral bleeding complications were assessed on follow-up magnetic resonance imaging or cranial computed tomography. Data were compared between patients <80 years of age and patients aged > or =80 years. RESULTS: A total of 228 patients were treated with IV-tPA; 38 (16%) were 80 years or older. There was no difference in NIHSS on admission or onset to treatment time between younger and older patients. Less patients > or =80 years of age achieved a favorable outcome (26.3 versus 46.8%, P=0.021), and mortality was higher in older patients (21.1 versus 5.3%, P=0.004). There was no difference in the rate of parenchymal hemorrhage (6.3%<80 years versus 5.3%> or =80 years, P=1.000) and symptomatic intracerebral hemorrhage (2.6%<80 years versus 2.6%> or =80 years, P=1.000) between both groups. CONCLUSIONS: There is no increase in severe intracerebral hemorrhage after IV-tPA in very old patients, but outcome is worse as compared with younger patients. There is no evidence to exclude ischemic stroke patients from thrombolysis based on a predefined age threshold. PMID- 16210555 TI - Apolipoprotein E genotype and incident ischemic stroke: the Atherosclerosis Risk in Communities Study. AB - BACKGROUND AND PURPOSE: A relationship between the apolipoprotein E (apoE) genotype and ischemic stroke has been inconsistently reported. We explored this relation in the Atherosclerosis Risk in Communities Study (ARIC). METHODS: The ARIC cohort involves 15 792 men and women, aged 45 to 64 years at baseline and sampled from 4 U.S. communities. Between 1987 and 2001, 498 incident ischemic strokes occurred. RESULTS: After stratifying by race and sex and adjusting for other nonlipid risk factors, there was no significant relation between the apoE genotype and incident stroke, except in black women (hazard ratio for epsilon2 genotype relative to epsilon3/epsilon3=0.53; 95% CI, 0.28 to 0.99). CONCLUSIONS: For the most part, in this middle-aged sample, apoE was not a risk factor for incident ischemic stroke. PMID- 16210556 TI - Incidence of recurrent subarachnoid hemorrhage after clipping for ruptured intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Because intracranial aneurysms develop during life, patients with subarachnoid hemorrhage (SAH) and successfully occluded aneurysms are at risk for a recurrence. We studied the incidence of and risk factors for recurrent SAH in patients who regained independence after SAH and in whom all aneurysms were occluded by means of clipping. METHODS: From a cohort of patients with SAH admitted between 1985 and 2001, we included those patients who were discharged home or to a rehabilitation facility. We interviewed these patients about new episodes of SAH. We retrieved all medical records and radiographs in case of reported recurrences. If patients had died, we retrieved the cause of death. We analyzed the incidence of and risk factors for recurrent SAH by Kaplan Meier curves and Cox regression analysis. RESULTS: Of 752 patients with 6016 follow-up years (mean follow up 8.0 years), 18 had a recurrence. In the first 10 years after the initial SAH, the cumulative incidence of recurrent SAH was 3.2% (95% confidence interval [CI], 1.5% to 4.9%) and the incidence rate 286 of 100,000 patient-years (95% CI, 160 to 472 per 100,000). Risk factors were smoking (hazard ratio [HR], 6.5; 95% CI, 1.7 to 24.0), age (HR, 0.5 per 10 years; 95% CI, 0.3 to 0.8) and multiple aneurysms at the time of the initial SAH (HR, 5.5; 95% CI, 2.2 to 14.1). CONCLUSIONS: After SAH, the incidence of a recurrence within the first 10 years is 22 (12 to 38) times higher than expected in populations with comparable age and sex. Whether this increased risk justifies screening for recurrent aneurysms in patients with a history of SAH requires further study. PMID- 16210558 TI - When a parent has a stroke: clinical course and prediction of mood, behavior problems, and health status of their young children. AB - BACKGROUND AND PURPOSE: The purpose of this research was to describe the clinical course of children's functioning (depression, behavioral problems, and health status) during the first year after parental stroke and to determine which patient-, spouse-, or child-related factors at the start of inpatient rehabilitation can predict children's functioning after parental stroke at 1-year poststroke. METHODS: Interviews with 82 children (4 to 18 years of age) and their parents (n=55) shortly after admission to a rehabilitation center, 2 months after discharge from inpatient rehabilitation, and 1 year after stroke. Depression was assessed using the Children Depression Inventory, behavioral problems with the Child Behavior Check List, and health status with the Functional Status II. Potential predictors were gender and age (child), activities of daily living disability and communication ability (patient), and spouse's depression and perception of the marital relationship. RESULTS: At the start of the stroke patient's rehabilitation, 54% of the children had > or =1 subclinical or clinical problems, which improved to 29% 1 year after stroke. Children's functioning 1 year after stroke could best be predicted by their functioning at the start of rehabilitation. Spouse depression and perception of marital relationship were also significant predictors. A total of 28% to 58% of the variance in children's functioning could be explained. CONCLUSIONS: Children's functioning after parental stroke improved during the first year after stroke. Identifying children at risk for problems 1 year after stroke requires assessment of children's functioning and the healthy spouse's depressive symptoms and perception of the marital relationship at the start of rehabilitation. This demonstrates the need for a family-centered approach in stroke rehabilitation. PMID- 16210557 TI - Electrocardiographic left atrial abnormalities and risk of ischemic stroke. AB - BACKGROUND AND PURPOSE: We evaluated the association between electrocardiographic left atrial abnormality (ECG-LAA) and ischemic stroke, especially whether ECG-LAA provides additional prognostic information to that provided by echocardiography. METHODS: A population-based, case-control study included 146 patients with first ischemic stroke and 195 age-, gender-, and race/ethnicity-matched community control subjects. ECG-LAA was defined as either P-wave duration >120 ms or P terminal force in precordial lead V1 (PTFV1) >40 ms.mm. RESULTS: PTFV1 >40 ms.mm was associated with ischemic stroke after adjustment for other stroke risk factors (odds ratio [OR], 2.32; 95% CI, 1.29 to 4.18). The association remained significant after adding echocardiographic left atrial diameter to the model (OR, 2.31; 95% CI, 1.28 to 4.17). PTFV1 was independently associated with stroke in patients in the upper half of echocardiographically determined left ventricular mass (adjusted OR, 4.5; 95% CI, 2.20 to 9.15) but not in those in the lower half (OR, 0.58; 95% CI, 0.20 to 1.65; P=0.0008). CONCLUSIONS: ECG-LAA can supplement 2D echocardiography in assessing the risk of ischemic stroke, especially in subjects with increased left ventricular mass. PMID- 16210559 TI - Silent cerebral ischemia detected with diffusion-weighted imaging in patients treated with protected and unprotected carotid artery stenting. AB - BACKGROUND AND PURPOSE: Percutaneous transluminal angioplasty with stent (CAS) is an alternative method to endarterectomy in the revascularization of carotid artery stenosis. Protected CAS is currently used to prevent distal embolization. Diffusion-weighted MRI (DWI) is the most sensitive tool to evaluate silent cerebral ischemia. The purpose of this research was to assess the incidence of cerebral embolic lesions during CAS and to evaluate whether cerebral protection devices can reduce the number of silent cerebral ischemia with respect to unprotected CAS. METHODS: Fifty-two patients with high-grade internal stenosis underwent CAS; 30 patients (group a) were treated with a cerebral protection device, and 22 (group b) were treated without it. All of the patients were evaluated preoperatively and postoperatively with fluid-attenuated inversion recovery and DWI sequences to depict the number of new embolic silent cerebral lesions. RESULTS: Embolic silent cerebral lesions occurred in 30% of CAS. Cerebral protection devices reduce the number of new lesions significantly reducing the consistent lesions ipsilateral to the treated vessel. Inconsistent lesions do not differ in both groups of patients. Clinical, radiological, and procedural variables do not correlate with the appearance of new cerebral lesions. CONCLUSIONS: Embolic cerebral lesions detected with DWI are more frequent with unprotected CAS, although they are present also with the use of cerebral protection devices. Probably a part of silent cerebral lesions arise from the procedural maneuver in the aortic arch. PMID- 16210560 TI - In situ beta-irradiation of a brain arteriovenous malformation model. AB - BACKGROUND AND PURPOSE: The treatment of large brain arteriovenous malformations (BAVMs) is challenging, and embolization alone is seldom curative. The study goal is to enhance the efficacy of arteriovenous malformation embolization by adding a beta-emitting isotope to the embolic agent. METHODS: The pig rete mirabile was used as a BAVM model. The body distribution of radioactivity was evaluated after selective rete injection of N-butyl,2-cyanoacrylate mixed with (131)I-lipiodol in 8 animals using immediate whole body gamma-scintigraphy. Activities within the whole rete mirabile and selected tissue samples were quantified with a gamma counter immediately after sacrifice. Two pigs were submitted to serial gamma scintigraphies for 6 weeks to detect delayed isotope leaching. Long-term effects of in situ irradiation were evaluated using a mixture of 188Re/N-butyl,2 cyanoacrylate in 8 pigs. In 1 animal, autoradiography was performed to evaluate local rete mirabile distribution of the radioactivity. Seven pigs were injected with 188Re/glue in 1 rete mirabile and with glue only on the opposite side, and the degree of vascular occlusion of both sides was compared on histology at 2 (n=2) or 6 months (n=5). RESULTS: There was negligible activity outside the target. Radiation caused occlusion of vessels unreached by the glue itself but in the vicinity of the radioactive source in 5 of 7 rete mirabile. CONCLUSIONS: Selective deposition of a beta-emitter inside a BAVM model may be achieved by current embolization techniques. The adjunct use of an isotope may increase the efficacy of embolization. PMID- 16210561 TI - Nedd9 protein, a Cas-L homologue, is upregulated after transient global ischemia in rats: possible involvement of Nedd9 in the differentiation of neurons after ischemia. AB - BACKGROUND AND PURPOSE: Some proteins involved in self-repair after stroke in the adult brain are primarily expressed during embryonic development and strongly down-regulated during the early postnatal phase. Neuronal precursor cell expressed, developmentally down-regulated gene (Nedd) 9 was recognized to be identical to Crk-associated substrate lymphocyte type (Cas-L), a docking protein that associates with a variety of signaling molecules, such as focal adhesion kinase (FAK), proline-rich tyrosine kinase 2 (Pyk2), and Crk. We investigated the involvement of these proteins in the pathophysiology of global cerebral ischemia. METHODS: The mouse Cas-L/Nedd9 cDNAs were cloned. The expression and function of Cas-L/Nedd9 protein in the pathogenesis of global ischemia in rats was investigated by RT-PCR, Western blot analysis, and immunohistochemistry. The neurite outgrowth of the transfectants of Nedd9 deletion mutants in PC-12 cells was also assessed to clarify the function of the Nedd9 protein. RESULTS: Nedd9 was a splicing variant of Cas-L and was selectively induced in neurons of the cerebral cortex and hippocampus 1 to 14 days after the ischemia. Induced Nedd9 protein was tyrosine phosphorylated and was bound to FAK in dendrite and soma of neurons after the ischemia. Finally, it was demonstrated that Nedd9 promoted neurite outgrowth of PC-12 cells. CONCLUSIONS: Our study may support the potential of Nedd9 for participation in the differentiation of neurons after global ischemia in rats. PMID- 16210562 TI - Short-term administration of a new free radical scavenger, edaravone, is more effective than its long-term administration for the treatment of neonatal hypoxic ischemic encephalopathy. AB - BACKGROUND AND PURPOSE: Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) is a new free radical scavenger that is used for the treatment of adult acute cerebral infarction in Japan. We examined the effect of edaravone on the optimal duration of treatment, the long-term effect on the brain, and the effect on learning and memory disability in a rat model of neonatal hypoxic-ischemic encephalopathy. METHODS: Seven-day-old Wistar rats were subjected to left common carotid artery ligation then 2 hours of hypoxic-ischemic insult or sham operation. Edaravone was administered intraperitoneally (9 mg/kg) after hypoxic-ischemic insult every 24 hours for 2, 5, or 10 consecutive days. The neuroprotective effect of edaravone was evaluated by behavioral test and histological analysis. RESULTS: Two-day treatment with edaravone significantly gave protection to the learning and memory capability, as well as morphological recovery compared with control rats. Five day treatment showed morphological improvement but no behavioral improvement. In contrast, 10-day treatment did not show either morphological or behavior improvement. CONCLUSIONS: These findings indicate that edaravone is a promising candidate as a treatment of choice for neonatal hypoxic-ischemic encephalopathy, when its use is limited to the acute phase after hypoxia-ischemia. PMID- 16210563 TI - Transcranial Doppler and carotid artery disease strokes: more than just risk stratification. PMID- 16210564 TI - A novel inhibitor of oxidosqualene:lanosterol cyclase inhibits very low-density lipoprotein apolipoprotein B100 (apoB100) production and enhances low-density lipoprotein apoB100 catabolism through marked reduction in hepatic cholesterol content. AB - OBJECTIVE: Inhibition of 2,3-oxidosqualene:lanosterol cyclase (OSC), an enzyme in the cholesterol synthesis pathway, has the unique ability to inhibit cholesterol synthesis while simultaneously enhancing oxysterol synthesis. Our objectives were to determine, in vivo, if a novel OSC inhibitor reduced low-density lipoprotein (LDL) cholesterol and to define the mechanism(s) involved. METHODS AND RESULTS: Miniature pigs received the OSC inhibitor RO0717625 or placebo and a diet containing fat (34% of energy) and 400 mg per day of cholesterol. Treatment decreased plasma total cholesterol (-20%) and LDL cholesterol (-29%). Apolipoprotein B (apoB) kinetic parameters were determined. Very low-density lipoprotein (VLDL) apoB pool size decreased 22% because of inhibition of VLDL production (-43%). LDL apoB pool size decreased 22% because of a 1.5-fold increase in fractional catabolic rate (FCR). The increased FCR was associated with a 2-fold increase in hepatic LDL receptor mRNA. Hepatic total and microsomal cholesterol were reduced by 16% and 27%, respectively. Plasma lathosterol concentrations decreased 57%, reflecting inhibition of hepatic cholesterol synthesis. Treatment reduced plasma plant sterols and decreased postprandial cholesterol transport in chylomicrons. CONCLUSIONS: A novel OSC inhibitor, RO0717625, decreased VLDL and LDL apoB100 through decreased VLDL production and enhanced LDL clearance. Thus, OSC represents a potential therapeutic target for dyslipidemia. PMID- 16210566 TI - Stimulated tissue plasminogen activator release as a marker of endothelial function in humans. AB - The initiation, modulation, and resolution of thrombus associated with eroded or unstable coronary plaques are critically dependent on the efficacy of endogenous fibrinolysis. This is dependent on the cellular function of the surrounding endothelium and vascular wall. In particular, the acute release of tissue plasminogen activator from the endothelium makes an important contribution to the defense against intravascular thrombosis. Here, we describe the rationale and methodology for, and clinical relevance of, assessing acute endothelial tissue plasminogen activator release in humans. The investigation of endothelial fibrinolytic function has the potential to provide major new insights into the pathophysiology of cardiovascular disease, and to shape future therapeutic interventions. PMID- 16210565 TI - Hyperhomocystinemia impairs endothelial function and eNOS activity via PKC activation. AB - OBJECTIVE: A risk factor for cardiovascular disease, hyperhomocystinemia (HHcy), is associated with endothelial dysfunction. In this study, we examined the mechanistic role of HHcy in endothelial dysfunction. METHODS AND RESULTS: Through the use of 2 functional models, aortic rings and intravital video microscopy of the cremaster, we found that arterial relaxation in response to the endothelium dependent vessel relaxant, acetylcholine or the nitric oxide synthase (NOS) activator (A23187), was significantly impaired in cystathionine beta-synthase null (CBS(-/-)) mice. However, the vascular smooth muscle cell (VSMC) response to the nitric oxide (NO) donor (SNAP) was preserved in CBS(-/-) mice. In addition, superoxide dismutase and catalase failed to restore endothelium-dependent vasodilatation. Endothelial nitric oxide synthase (eNOS) activity was significantly reduced in mouse aortic endothelial cells (MAECs) of CBS(-/-) mice, as well as in Hcy-treated mouse and human aortic endothelial cells (HAECs). Hcy mediated eNOS inhibition--which was not rescued by adenoviral transduction of superoxide dismutase and glutathione peroxidase, or by tetrahydrobiopterin, sepiapterin, and arginine supplementations in MAEC--was associated with decreased protein expression and increased threonine 495 phosphorylation of eNOS in HAECs. Ultimately, a protein kinase C (PKC) inhibitor, GF109203X (GFX), reversed Hcy mediated eNOS inactivation and threonine 495 phosphorylation in HAECs. CONCLUSIONS: These data suggest that HHcy impairs endothelial function and eNOS activity, primarily through PKC activation. PMID- 16210567 TI - Effect of nuclear actin on endothelial nitric oxide synthase expression. AB - BACKGROUND: Previously, we showed that the 27nt repeat polymorphism in endothelial nitric oxide synthase (eNOS) intron 4 was associated with altered eNOS mRNA and protein levels, nitric oxide (NO) production and vascular disease risk; the 27-nt repeats had a cis-acting role in eNOS promoter function. In the present study, we investigated nuclear protein that binds the 27nt repeat and mediates eNOS expression. METHODS AND RESULTS: Using 5'-biotin-labeled 27nt DNA duplex and streptavidin-agarose beads pull-down assay and mass spectrometry, we identified that nuclear beta-actin was one of the major 27nt binding proteins. Using the pGL3 reporter vectors containing the 5x27nt repeats as an enhancer in an in vitro transcription assay, we found that exogenous beta-actin significantly increased reporter gene transcription efficiency. The beta-actin's upregulating effect was compromised when exogenous 27nt RNA duplex was added. Furthermore, the eNOS expression was reduced when beta-actin gene was silenced by specific siRNA, and actin overexpression upregulated eNOS expression >3-fold. CONCLUSIONS: Our data demonstrate that beta-actin as a transcription factor stimulates eNOS expression; and the transcriptional effect appears to be 27nt-dependent. Our findings represent a novel molecular mechanism regulating eNOS expression, which could potentially lead to discoveries of eNOS specific pharmaceutical agents, eg, active peptides, with clinical applications. PMID- 16210568 TI - Fibrinogen contains cryptic PAI-1 binding sites that are exposed on binding to solid surfaces or limited proteolysis. AB - OBJECTIVE: In this work, we identified the fibrinogen sequence that on exposure serves as the primary binding site for functionally active PAI-1 and to a lesser extent for its latent form. In contrast, this site only weakly interacts with PAI 1 substrate. METHODS AND RESULTS: The binding site is located in the N-terminal alpha (20-88) segment of fibrinogen, in the region exposed on (1) adsorption of fibrinogen to solid surfaces; (2) the release of fibrinopeptide A during thrombin conversion of fibrinogen to fibrin; and (3) plasmin degradation of fibrinogen. This region was first identified by the yeast 2-hybrid system, then its binding characteristics were evaluated using the recombinant alpha(16-120) fragment and its shorter version, the alpha(20-88) fragment, in a solid phase binding assay and plasmon surface resonance measurements. Because fibrinogen fragment E does not bind PAI-1, it suggests that sequences of Aalpha chain interacting with PAI-1 are located in the N-terminal part of the alpha(20-88) segment. CONCLUSIONS: Therefore, PAI-1 directly bound to the alpha(20-88) and thus concentrated in fibrinogen/fibrin, particularly at sites of injury and inflammation, may account for the recent observations that both its active and latent forms stimulate cell migration and wound healing. PMID- 16210570 TI - Upregulation of proinflammatory proteins through NF-kappaB pathway by shed membrane microparticles results in vascular hyporeactivity. AB - OBJECTIVE: Microparticles are membrane vesicles with procoagulant and proinflammatory properties released during cell activation, including apoptosis. The present study was designed in dissecting the effects evoked by microparticles on vascular reactivity. METHODS AND RESULTS: Microparticles from either apoptotic T lymphocytic cells or from plasma of diabetic patients with vascular complications induced vascular hyporeactivity in response to vasoconstrictor agents in mouse aorta. Hyporeactivity was reversed by nitric oxide (NO) synthase plus cyclooxygenase-2 inhibitors, and associated with an increased production of vasodilatory products such as NO and prostacyclin. Microparticles induced an upregulation of proinflammatory protein expressions, inducible NO-synthase and cyclooxygenase-2, mainly in the medial layer of the vessels as evidenced by immunochemical staining. In addition, microparticles evoke NF-kappaB activation probably through the interaction with the Fas/Fas Ligand pathway. Finally, in vivo treatment of mice with lymphocyte-derived MPs induces vascular hyporeactivity, which was reversed by the combination of NO and cyclooxygenase-2 inhibitors. CONCLUSIONS: These data provide a rationale to explain the paracrine role of microparticles as vectors of transcellular exchange of message in promoting vascular dysfunction during inflammatory diseases. PMID- 16210569 TI - Smoking, endothelial function, and Rho-kinase in humans. AB - OBJECTIVE: Smoking is associated with endothelial dysfunction and activated Rho kinase in vascular smooth muscle cells (VSMCs) in humans. The purpose of this study was to elucidate the relationship between endothelial function and Rho kinase activity in forearm VSMCs in healthy young men. METHODS AND RESULTS: We evaluated the forearm blood flow (FBF) responses to acetylcholine (ACh), fasudil, a Rho-kinase inhibitor, and sodium nitroprusside (SNP) in male smokers (n=10) and nonsmokers (n=14). FBF was measured by using a strain-gauge plethysmography. The vasodilatory effect of ACh was significantly smaller in smokers than that in nonsmokers. The vasodilatory effect of fasudil was significantly greater in smokers than that in nonsmokers. The vasodilatory effects of SNP in the 2 groups were similar. There was a significant correlation between the maximal FBF response to fasudil and that to ACh (r=-0.67; P<0.01). There was no significant correlation between the maximal FBF response to fasudil and that to SNP. The intra-arterial coinfusion of fasudil significantly increased the FBF response to ACh in smokers but not in nonsmokers. There were no significant differences between FBF response to fasudil alone and that in combination with NG-monomethyl L-arginine in smokers and in nonsmokers. The intra-arterial coinfusion ascorbic acid did not alter the FBF response to fasudil in both groups. CONCLUSIONS: These findings suggest that smoking is involved in not only endothelial dysfunction but also activation of Rho-kinase in VSMCs in forearm circulation, and that there is a significant correlation between endothelial function and Rho-kinase activity in VSMCs. PMID- 16210571 TI - Peroxynitrite causes endoplasmic reticulum stress and apoptosis in human vascular endothelium: implications in atherogenesis. AB - OBJECTIVE: Peroxynitrite, a potent oxidant generated by the reaction of NO with superoxide, has been implicated in the promotion of atherosclerosis. We designed this study to determine whether peroxynitrite induces its proatherogenic effects through induction of endoplasmic reticulum (ER) stress. METHODS AND RESULTS: Human vascular endothelial cells treated with Sin-1, a peroxynitrite generator, induced the expression of the ER chaperones GRP78 and GRP94 and increased eIF2alpha phosphorylation. These effects were inhibited by the peroxynitrite scavenger uric acid. Sin-1 caused the depletion of ER-Ca2+, an effect known to induce ER stress, resulting in the elevation of cytosolic Ca2+ and programmed cell death (PCD). Sin-1 treatment was also found, via 3-nitrotyrosine and GRP78 colocalization, to act directly on the ER. Adenoviral-mediated overexpression of GRP78 in endothelial cells prevented Sin-1-induced PCD. Consistent with these in vitro findings, 3-nitrotyrosine was observed and colocalized with GRP78 in endothelial cells of early atherosclerotic lesions from apolipoprotein E deficient mice. CONCLUSIONS: Peroxynitrite is an ER stress-inducing agent. Its effects include the depletion of ER-Ca2+, a known mechanism of ER stress induction. The observation that 3-nitrotyrosine-containing proteins colocalize with markers of ER stress within early atherosclerotic lesions suggests that peroxynitrite contributes to atherogenesis through a mechanism involving ER stress. PMID- 16210572 TI - Chlamydia pneumoniae burden in carotid arteries is associated with upregulation of plaque interleukin-6 and elevated C-reactive protein in serum. AB - OBJECTIVE: Chlamydia pneumoniae (Cpn) infection of vascular smooth muscle cells increases interleukin-6 (IL-6) secretion in vitro. In vivo, IL-6 stimulates liver C-reactive protein (CRP) production. Because serum levels of IL-6 and CRP are independent risk factors for stroke and myocardial infarction (MI), we investigated whether Cpn burden in carotid plaques might provide a link between plaque IL-6 expression and elevated serum levels of IL-6 and CRP. METHODS AND RESULTS: Consecutive patients undergoing elective carotid endarterectomy were studied. Serum levels of CRP and IL-6 were measured before surgery. Immunohistochemistry and real-time quantitative (k)RT-PCR were used to detect Cpn and the expression of IL-6 within carotid plaques. Cpn mRNA was present in 19 (37%) of 51 patients, suggesting viable infections. These patients had evidence for infection by PCR and immunohistochemistry. The Cpn burden, measured by real time quantitative (k)-PCR using the number of organisms normalized against the number of eukaryotic cells in the tissue, was associated with plaque expression of IL-6 (Spearman R=0.55; P<0.0001), which was associated with serum levels of IL 6 (R=0.56; P<0.0001) and CRP (R=0.80; P<0.0001). CONCLUSIONS: IL-6 secretion in atherosclerotic plaques infected with Cpn could explain elevated serum inflammatory markers in individuals at risk for stroke and MI. PMID- 16210573 TI - Indomethacin and celecoxib impair rotator cuff tendon-to-bone healing. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs are commonly prescribed after rotator cuff repair. These agents can impair bone formation, but no studies have evaluated their impact on tendon-to-bone healing. HYPOTHESIS: Traditional nonselective nonsteroidal anti-inflammatory drugs and cyclooxygenase-2-specific nonsteroidal anti-inflammatory drugs interfere with tendon-to-bone healing. STUDY DESIGN: Controlled laboratory study. METHODS: One hundred eighty Sprague-Dawley rats underwent acute rotator cuff repairs. Postoperatively, 60 rats received 14 days of celecoxib, a cyclooxygenase-2-specific nonsteroidal anti-inflammatory drug; 60 received indomethacin, a traditional nonselective nonsteroidal anti inflammatory drug; and 60 received standard rat chow. Animals were sacrificed at 2, 4, and 8 weeks and evaluated by gross inspection, biomechanical testing, histologic analysis, and polarized light microscopy to quantify collagen formation and maturation. RESULTS: Five tendons completely failed to heal (4 celecoxib, 1 indomethacin). There were significantly lower failure loads in the celecoxib and indomethacin groups compared with the control groups at 2, 4, and 8 weeks (P < .001), with no significant difference between nonsteroidal anti inflammatory drug groups. There were significant differences in collagen organization and maturation between the controls and both nonsteroidal anti inflammatory drug groups at 4 and 8 weeks (P < .001). Controls demonstrated progressively increasing collagen organization during the course of the study (P < .001), whereas the nonsteroidal anti-inflammatory drug groups did not. CONCLUSION: Traditional and cyclooxygenase-2-specific nonsteroidal anti inflammatory drugs significantly inhibited tendon-to-bone healing. This inhibition appears linked to cyclooxygenase-2. CLINICAL RELEVANCE: If the results of this study are verified in a larger animal model, the common practice of administering non-steroidal anti-inflammatory drugs after rotator cuff repair should be reconsidered. PMID- 16210574 TI - Effect of gender and maturity on quadriceps-to-hamstring strength ratio and anterior cruciate ligament laxity. AB - BACKGROUND: Exercise programs have been introduced to reduce the ACL injury risk in female athletes. The most effective age at which to start these programs is not known. HYPOTHESIS: Age and gender affect ligament laxity and quadriceps-to hamstring strength ratio. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: Fifty-three female and 70 male recreational soccer players, 10 to 18 years of age, were studied with physical examination, KT-1000 arthrometry, and manual maximum quadriceps and hamstring strength using a handheld dynamometer. The subjects were separated into 4 groups to examine maturity-related intergender differences: group G1, premenarchal girls (n = 24); group B1, boys 13 years and younger (n = 38); group G2, girls 2 or more years after menarche (n = 29); and group B2, boys 14 years and older (n = 32). RESULTS: Both knees of 123 soccer players were evaluated. The mean ages for groups G1, B1, G2, and B2 were 11.50 +/ 1.69, 10.63 +/- 1.85, 15.5 +/- 1.43, and 15.59 +/- 1.24 years, respectively, and the mean laxity measurements were 8.84 +/- 2.12, 8.51 +/- 1.61, 8.85 +/- 1.86, and 7.33 +/- 1.27 mm, respectively. Laxity was significantly less for the mature boys (P = .0015) than for the immature boys, mature girls, and immature girls. With increasing maturity, significant increases in both quadriceps and hamstring muscle strength were observed for both boys and girls (P < .05). Boys demonstrated a greater percentage increase in hamstring strength with maturity (179%) compared with girls (27%) (P < .05). Mature girls (2.06) had significantly greater quadriceps-to-hamstring ratio when compared with immature girls (1.74), immature boys (1.58), and mature boys (1.48) (P < .05). CONCLUSION: Female athletes after menarche increase their quadriceps strength greater than their hamstring strength, putting them at risk for anterior cruciate ligament injury. Anterior cruciate ligament-prevention programs based on improving dynamic control of the knee by emphasizing hamstring strengthening should be instituted for girls after menarche. PMID- 16210575 TI - Femoral press-fit fixation technique in anterior cruciate ligament reconstruction using bone-patellar tendon-bone graft: a prospective clinical evaluation of 285 patients. AB - BACKGROUND: The security of the graft fixation is an important factor of anterior cruciate ligament reconstruction, especially in the early postoperative period. HYPOTHESIS: The femoral press-fit fixation technique is a safe, alternative technique for the fixation of the bone-patellar tendon-bone graft. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Between January 1, 1998, and June 30, 2002, 285 patients, with a mean age of 29.1 years, underwent operation and observation in our department. All patients had arthroscopic anterior cruciate ligament reconstruction using bone-patellar tendon-bone graft with femoral press fit and tibial interference screw fixation. The mean follow-up period was 35.8 months (range, 24-77 months). During each patient's evaluation, a physical examination, a subjective evaluation, the International Knee Documentation Committee and Lysholm scores, KT-1000 arthrometer testing, and radiological evaluation were performed. The paired Student t test or chi(2) test was used for statistical analysis. RESULTS: According to the International Knee Documentation Committee score, 241 patients (85%) had good (46 normal and 195 nearly normal) results, whereas 39 patients had abnormal and 5 patients had severely abnormal overall results. The preoperative mean Lysholm score was 63.5 +/- 12.7 points. At the final examination, the mean value was 93.5 +/- 7.8 points (P < .01). One hundred ninety-seven athletes could return to their previous sports activity level, whereas 13 patients gave up their sports career. The KT-1000 arthrometer results showed a side-to-side difference of 1.91 +/- 2.1 mm at the maximum manual strength tests. CONCLUSION: The femoral press-fit fixation is an alternative fixation method for the bone-patellar tendon-bone graft and provides good stability for the anterior cruciate ligament reconstruction. The use of press-fit fixation technique avoids most interference screw or other hardware-induced complications at the femoral side. PMID- 16210576 TI - Acute cervical spine injuries in mountain biking: a report of 3 cases. PMID- 16210577 TI - The effects of augmentation with Swine small intestine submucosa on tendon healing under tension: histologic and mechanical evaluations in sheep. AB - BACKGROUND: Rotator cuff failure after surgery may be attributed to inferior tissue healing properties that result from repetitive cyclic loading during early rehabilitation. Enhancing the biological healing process may reduce the incidence of failures after rotator cuff repairs. HYPOTHESIS: Augmentation of rotator cuff tissue using swine small intestine submucosa in a sheep model will improve the rate and quality of tissue repair. STUDY DESIGN: Controlled laboratory study. METHODS: We resected and reattached 26 sheep infraspinatus tendons under tension, with 13 animals receiving a small intestine submucosa patch (augmented group). Animals were sacrificed at 12 weeks, and biomechanical testing and histologic evaluation were performed. Biomechanical testing was completed in 10 tendons from each group. Specimens were loaded to failure at a constant displacement to obtain the load deformation curve used to calculate load to failure and stiffness of the healed bone-tendon interface. Histologic testing addressed tissue healing at the bone-tendon interface. RESULTS: The load-to-failure data did not indicate a significant difference between the augmented and nonaugmented groups (1252 +/- 402 N vs 985 +/- 459 N, respectively; P > .05). However, the augmented group had significantly better stiffness than the nonaugmented group (215 +/- 44 N/mm vs 154 +/- 63 N/mm, respectively; P = .03). Histologic data revealed that the infraspinatus tendon in all specimens inserted into the bone through a zone of fibrocartilage, although none of the patches were intact. CONCLUSION: Although there were no differences in the load-to-failure data between the 2 groups, the statistically significant improvement in stiffness for the augmented group is clinically relevant. Stiffness is the biomechanical parameter representing the tissue response to subdestructive loads seen with early rehabilitation. Augmenting the repair with a collagen matrix improved the early healing characteristics of the repair construct. CLINICAL RELEVANCE: Enhancing the biological process of tendon healing under tension by using a collagen matrix patch may improve the ultimate success of rotator cuff repair. PMID- 16210578 TI - Arthroscopic single-row versus double-row suture anchor rotator cuff repair. AB - BACKGROUND: Recurrent defects after open and arthroscopic rotator cuff repair are common. Double-row repair techniques may improve initial fixation and quality of rotator cuff repair. PURPOSE: To evaluate the load to failure, cyclic displacement, and anatomical footprint of 4 arthroscopic rotator cuff repair techniques. HYPOTHESIS: Double-row suture anchor repair would have superior structural properties and would create a larger footprint compared to single-row repair. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty fresh-frozen cadaveric shoulders were randomly assigned to 4 arthroscopic repair techniques. The repair was performed as either a single-row technique or 1 of 3 double-row techniques: diamond, mattress double anchor, or modified mattress double anchor. Angle of loading, anchor type, bone mineral density, anchor distribution, angle of anchor insertion, arthroscopic technique, and suture type and size were all controlled. Footprint length and width were quantified before and after repair. Displacement with cyclic loading and load to failure were determined. RESULTS: There were no differences in load to failure and displacement with cyclic loading between the single-row repair and each double-row repair. All repair groups demonstrated load to failure greater than 250 N. A significantly greater supraspinatus footprint width was seen with double-row techniques compared to single-row repair. CONCLUSIONS: The single-row repair technique was similar to the double-row techniques in load to failure, cyclic displacement, and gap formation. The double-row anchor repairs consistently restored a larger footprint than did the single-row method. CLINICAL RELEVANCE: The arthroscopic techniques studied have strong structural properties that approached the reported performance of open repair techniques. Double-row techniques provide a larger footprint width; although not addressed by this study, such a factor may improve the biological quality of repair. PMID- 16210579 TI - Hamstring muscle function after tendon harvest for anterior cruciate ligament reconstruction: evaluation with T2 relaxation time of magnetic resonance imaging. AB - BACKGROUND: Regeneration of the semitendinosus and gracilis tendons after harvesting for anterior cruciate ligament reconstruction has been reported; however, muscle belly function after tendon regeneration has not been well documented. HYPOTHESIS: The semitendinosus and gracilis muscles are highly activated during knee flexion if their tendons are well regenerated after anterior cruciate ligament reconstruction. STUDY DESIGN: Descriptive laboratory study. METHODS: Hamstring muscle activation in 11 patients who had undergone anterior cruciate ligament reconstruction with semitendinosus and gracilis tendons was evaluated by measuring the increase of T2 relaxation time measured via magnetic resonance imaging after knee flexion exercise. Tendon regeneration was evaluated via magnetic resonance imaging. RESULTS: Both muscles increased T2 relaxation time after knee flexion exercise in the operated legs, and there was no significant difference in those values between the operated and nonoperated legs. All the semitendinosus tendons were regenerated at or below the joint line, but no gracilis tendons were observed beyond the joint line. The results indicated that both muscles were highly recruited during knee flexion, regardless of the degree of their tendon regeneration. CONCLUSION: The semitendinosus and gracilis muscles are able to restore or maintain their contractile capability after harvest of their tendons for anterior cruciate ligament reconstruction, regardless of the degree of regeneration. PMID- 16210580 TI - Effects of local administration of vascular endothelial growth factor on properties of the in situ frozen-thawed anterior cruciate ligament in rabbits. AB - BACKGROUND: In the autogenous tendon for anterior cruciate ligament reconstruction, intrinsic fibroblasts are necrotized immediately after surgery, and repopulation and revascularization occur. Vascular endothelial growth factor is considered to be a potent mediator of angiogenesis. HYPOTHESIS: An application of vascular endothelial growth factor significantly enhances angiogenesis in the in situ frozen anterior cruciate ligament, and the application significantly affects mechanical properties of the in situ frozen anterior cruciate ligament. STUDY DESIGN: Controlled laboratory study. METHODS: Right anterior cruciate ligaments from 66 rabbits underwent the freeze-thaw treatment, and animals were then divided into 3 groups. Group I served as a freeze-thaw but otherwise untreated control. In group II, 0.2 mL phosphate-buffered saline alone was applied. In group III, 30 mug vascular endothelial growth factor was applied. The groups were compared on the basis of histologic revascularization examinations using the Chalkley score, an indicator of the microvessel density, and mechanical evaluations, which included the anterior-posterior translation of the tibia relative to the femur during +/- 10 N of anterior-posterior load and the mechanical properties of the anteromedial bundle of the anterior cruciate ligament. RESULTS: Group III's Chalkley score was significantly greater than that of groups I and II. The tensile strength and the tangent modulus of anterior cruciate ligaments in groups I, II, and III were significantly lower than those of a normal anterior cruciate ligament, although there were no significant differences among groups I, II, and III. CONCLUSION: Vascular endothelial growth factor, as administered in this study, significantly promoted angiogenesis in the devitalized anterior cruciate ligament with in situ freeze-thaw treatment, but it did not affect the mechanical properties of the in situ frozen-thawed anterior cruciate ligament in the rabbit model. CLINICAL RELEVANCE: An application of the recombinant anterior cruciate ligament is a potential future strategy to enhance revascularization of the autograft in anterior cruciate ligament reconstruction. PMID- 16210581 TI - The effect of direction and reaction on the neuromuscular and biomechanical characteristics of the knee during tasks that simulate the noncontact anterior cruciate ligament injury mechanism. AB - BACKGROUND: Jumping and landing tasks that have a change in direction have been implicated as a mechanism of noncontact anterior cruciate ligament injury. Yet, to date, neuromuscular and biomechanical research has focused primarily on straight landing tasks during planned jumps. HYPOTHESIS: Lateral and reactive jumps increase the neuromuscular and biomechanical demands placed on the anterior cruciate ligament, and women perform these tasks differently from men. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 18 male and 17 female healthy high school basketball players underwent an analysis of the knee during planned and reactive 2-legged stop-jump tasks in 3 different directions that included novel methodology to incorporate a reactive component. Ground-reaction forces, joint kinematics, joint kinetics, and electromyographic activity were assessed during the tasks. RESULTS: Jump direction and task (planned or reactive) significantly affected joint angles, ground-reaction forces, knee joint moments, and proximal anterior tibia shear forces; female players demonstrated different kinematic, kinetic, and electromyographic characteristics during these tasks. CONCLUSION AND CLINICAL RELEVANCE: Jump direction significantly influenced knee biomechanics, suggesting that lateral jumps are the most dangerous of the stop jumps. Reactive jumps were also significantly different, suggesting differences between planned laboratory experiments and actual athletic competition. The results of this study indicate that directional and reactive jumps should be included in research methodology and injury-prevention programs. PMID- 16210582 TI - Thanks for the memory! PMID- 16210583 TI - Cutting edge: differential self-peptide/MHC requirement for maintaining CD8 T cell function versus homeostatic proliferation. AB - Memory T cells do not require self-peptide/MHC (spMHC) complexes to survive long term in vivo. However, memory CD4 T cells lose the ability to reject skin grafts when transiently placed in an environment in which these low-level TCR stimulations are absent. Whether or not spMHC alters the ability of CD8 T cells to respond to stimulation in vivo remains unknown. Here, we show that memory CD8 T cells retain the ability to respond to dendritic cell-mediated stimulation after adoptive transfer into either TAP(-/-) (MHC class I-deficient) or wild-type mice. Surprisingly, naive CD8 T cells, which fail to undergo homeostatic proliferation and erode in number in the absence of MHC class I, also retain the ability to respond to dendritic cell-mediated antigenic stimulation for at least 1 wk after transfer into TAP(-/-) mice. These findings suggest a differential requirement for spMHC signals for maintenance of CD8 T cell function and homeostatic proliferation. PMID- 16210584 TI - Cutting edge: TLR4 deficiency confers susceptibility to lethal oxidant lung injury. AB - TLRs have been studied extensively in pathogen-mediated host responses. We use a murine model of lethal oxidant-mediated injury to demonstrate for the first time that mammalian TLR4 is required for survival and lung integrity. Administering high levels of inspired oxygen, or hyperoxia, is commonly used as a life sustaining measure in critically ill patients. However, prolonged exposures can lead to respiratory failure and death. TLR4-deficient mice exhibited increased mortality and lung injury during hyperoxia. The enhanced susceptibility of TLR4 deficient mice to hyperoxia was associated with an inability to up-regulate Bcl-2 and phospho-Akt. Restoration of Bcl-2 and phospho-Akt levels by the exogenous transfer of the antioxidant gene heme oxygenase-1 markedly attenuated hyperoxia induced injury, apoptosis, and mortality in TLR4-deficient mice. Taken together, our results suggest a protective role of TLR4 in oxidant-mediated injury, providing novel mechanistic links among innate immunity, oxidant stress, and apoptosis. PMID- 16210585 TI - Gene family clustering identifies functionally associated subsets of human in vivo blood and tonsillar dendritic cells. AB - Human dendritic cells (DCs) are a distinct but heterogeneous lineage of APCs operating as the link between innate and adaptive immune responses, with the function to either maintain tolerance or trigger immunity. The DC lineage consists of several subpopulations with unique phenotypes; however, their functional characteristics and transcriptional similarities remain largely unknown. To further characterize the phenotypes and transcriptomes of the subsets, we purified myeloid CD16(+), blood DC Ag 1(+) (BDCA1(+)), and BDCA3(+) DC populations, as well as plasmacytoid CD123(+) DCs, from tonsillar tissue and peripheral blood. Transcriptional profiling and hierarchical clustering visualized that BDCA1(+) DCs clustered with BDCA3(+) DCs, whereas CD16(+) DCs and CD123(+) DCs clustered as distinct populations in blood. Differential expression levels of chemokines, ILs, and pattern recognition receptors were demonstrated, which emphasize innate DC subset specialization. Even though highly BDCA1(+) and BDCA3(+) DC-specific gene expression was identified in blood, the BDCA1(+) DCs and BDCA3(+) DCs from tonsils displayed similar transcriptional activity, most likely due to the pathogenic or inflammatory maturational signals present in tonsillar tissues. Of note, plasmacytoid DCs displayed less plasticity in their transcriptional activity compared with myeloid DCs. The data demonstrated a functionally distinct association of each of the seven subsets based on their signatures, involving regulatory genes in adaptive and innate immunity. PMID- 16210586 TI - Inefficient cell spreading and cytoskeletal polarization by CD4+CD8+ thymocytes: regulation by the thymic environment. AB - CD4(+)CD8(+) double-positive (DP) thymocytes express a lower level of surface TCR than do mature T cells or single-positive (SP) thymocytes. Regulation of the TCR on DP thymocytes appears to result from intrathymic signaling, as in vitro culture of these cells results in spontaneous TCR up-regulation. In this study, we examined cell spreading and cytoskeletal polarization responses that have been shown to occur in response to TCR engagement in mature T cells. Using DP thymocytes stimulated on lipid bilayers or nontransgenic thymocytes added to anti CD3-coated surfaces, we found that cell spreading and polarization of the microtubule organizing center and the actin cytoskeleton were inefficient in freshly isolated DP thymocytes, but were dramatically enhanced after overnight culture. SP (CD4(+)) thymocytes showed efficient responses to TCR engagement, suggesting that releasing DP thymocytes from the thymic environment mimics some aspects of positive selection. The poor translation of a TCR signal to cytoskeletal responses could limit the ability of DP thymocytes to form stable contacts with APCs and may thereby regulate thymocyte selection during T cell development. PMID- 16210588 TI - The CD85J/leukocyte inhibitory receptor-1 distinguishes between conformed and beta 2-microglobulin-free HLA-G molecules. AB - For a proper development of the placenta, maternal NK cells should not attack the fetal extravillous cytotrophoblast cells. This inhibition of maternal NK cells is partially mediated via the nonclassical MHC class I molecule HLA-G. Recently, we demonstrated that HLA-G forms disulfide-linked high molecular complexes on the surface of transfected cells. In the present study, we demonstrate that HLA-G must associate with beta(2)m for its interaction with CD85J/leukocyte Ig-like receptor-1 (LIR-1). Although HLA-G free H chain complexes are expressed on the surface, they are not recognized and possibly interfere with CD85J/LIR-1 and HLA G interaction. The formation of these complexes on the cell surface might represent a novel mechanism developed specifically by the HLA-G protein aimed to control the efficiency of the CD85J/LIR-1-mediated inhibition. We also show that endogenous HLA-G complexes are expressed on the cell surface. These findings provide novel insights into the delicate interaction between extravillous cytotrophoblast cells and NK cells in the decidua. PMID- 16210587 TI - Propensity of adult lymphoid progenitors to progress to DN2/3 stage thymocytes with Notch receptor ligation. AB - Notch family receptors control critical events in the production and replenishment of specialized cells in the immune system. However, it is unclear whether Notch signaling regulates abrupt binary lineage choices in homogeneous progenitors or has more gradual influence over multiple aspects of the process. A recently developed coculture system with Delta 1-transduced stromal cells is being extensively used to address such fundamental questions. Different from fetal progenitors, multiple types of adult marrow cells expanded indefinitely in murine Delta-like 1-transduced OP9 cell cocultures, progressed to a DN2/DN3 thymocyte stage, and slowly produced TCR(+) and NK cells. Long-term cultured cells of this kind retained some potential for T lymphopoiesis in vivo. Adult marrow progressed through double-positive and single-positive stages only when IL 7 concentrations were low and passages were infrequent. Lin(-)c-Kit(low)GFP(+)IL 7Ralpha(+/-) prolymphocytes were the most efficient of adult bone marrow cells in short-term cultures, but the assay does not necessarily reflect cells normally responsible for replenishing the adult thymus. Although marrow-derived progenitors with Ig D(H)-J(H) rearrangements acquired T lineage characteristics in this model, that was not the case for more B committed cells with V(H) D(H)J(H) rearrangement products. PMID- 16210589 TI - Protection from abortion by heme oxygenase-1 up-regulation is associated with increased levels of Bag-1 and neuropilin-1 at the fetal-maternal interface. AB - Tolerance mechanisms allowing pregnancy success resemble those involved in allograft acceptance. Heme oxygenase (HO) is a tissue-protective molecule, which allows graft acceptance and is known to have antiapoptotic effects on several cell types. We previously reported down-regulated levels of HO-1 and HO-2 in placenta from allopregnant mice undergoing abortion. In this study, we analyzed whether the up-regulation of HO-1 by cobalt-protoporphyrin (Co-PP) during implantation window can rescue mice from abortion. Induction of HO-1 by Co-PP treatment prevented fetal rejection, whereas the down-regulation of HOs by zinc protoporphyrin application boosted abortion. The beneficial effect of HO-1 induction was not related to a local shift to Th2-profile or to a change in the NO system. Interestingly, the expression of the antiapoptotic/cytoprotective molecule Bag-1 as well as the levels of neuropilin-1, a novel marker for T regulatory cells, were up-regulated after Co-PP treatment. Our data strongly support a very important role for HO-1 in fetal allotolerance and suggest that HO 1 might be protective by up-regulating tissue protective molecules, i.e., Bag-1, and by activating T regulatory cells rather than by changing the local cytokine profile. PMID- 16210590 TI - Memory T cells and their costimulators in human allograft injury. AB - Both CD4(+) and CD8(+) human memory but not naive T cells respond to allogeneic human dermal microvascular endothelial cells (HDMEC) in vitro by secreting cytokines and by proliferating. Several recently identified costimulators, namely, 4-1BB ligand, ICOS ligand, and OX40 ligand, are up-regulated on cultured HDMEC in response to TNF or coculture with allogeneic T cells. Blockade of these costimulators each partially reduces IFN-gamma and IL-2 secretion and proliferation of previously resting memory T cells. The effects of these costimulators are overlapping but not identical. Memory but not naive T cells are the principal effectors of microvascular injury in human skin allografts following adoptive transfer into immunodeficient mice. Furthermore, blocking 4 1BB ligand, ICOS ligand, or OX40 ligand in this model reduces human skin allograft injury and T cell effector molecule expression. These data demonstrate that human memory T cells respond to microvascular endothelial cells and can injure allografts in vivo without priming. Furthermore, several recently described costimulators contribute to these processes. PMID- 16210591 TI - Engagement of 4-1BB inhibits the development of experimental allergic conjunctivitis in mice. AB - The 4-1BB receptor acts as a costimulator in CD8(+) T cell activation. Agonistic stimulation through this molecule by treatment with anti-4-1BB Abs has been demonstrated to inhibit various experimentally induced diseases in animals. However, the effect of anti-4-1BB Abs on experimental allergic diseases has not been reported. We investigated the effect of anti-4-1BB Abs on the development and progression of experimental allergic conjunctivitis in mice. To examine the effects of Abs during the induction or effector phase, actively immunized mice or passively immunized mice by splenocyte transfer were treated with agonistic anti 4-1BB Abs, blocking anti-4-1BB ligand Abs, or normal rat IgG. Eosinophil infiltration into the conjunctiva was significantly reduced in wild-type mice by the anti-4-1BB Ab treatment during either induction or effector phase. Th2 cytokine production by splenocytes and total serum IgE were significantly reduced by the anti-4-1BB Ab treatment, while IFN-gamma production was increased. The anti-4-1BB Ab treatment induced a relative increase of CD8-positive cell numbers in the spleens. Moreover, inhibition of eosinophil infiltration by the treatment with anti-4-1BB Abs was also noted in actively immunized IFN-gamma knockout mice. Taken altogether, in vivo treatment with agonistic anti-4-1BB Abs in either induction or effector phase inhibits the development of experimental allergic conjunctivitis, and this inhibition is likely to be mediated by suppression of Th2 immune responses rather than up-regulation of IFN-gamma. PMID- 16210592 TI - Role of CXC chemokine ligand 13, CC chemokine ligand (CCL) 19, and CCL21 in the organization and function of nasal-associated lymphoid tissue. AB - Nasal-associated lymphoid tissue (NALT) orchestrates immune responses to Ags in the upper respiratory tract. Unlike other lymphoid organs, NALT develops independently of lymphotoxin-alpha (LTalpha). However, the structure and function of NALT are impaired in Ltalpha(-/-) mice, suggesting a link between LTalpha and chemokine expression. In this study we show that the expression of CXCL13, CCL19, CCL21, and CCL20 is impaired in the NALT of Ltalpha(-/-) mice. We also show that the NALT of Cxcl13(-/-) and plt/plt mice exhibits some, but not all, of the structural and functional defects observed in the NALT of Ltalpha(-/-) mice. Like the NALT of Ltalpha(-/-) mice, the NALT in Cxcl13(-/-) mice lacks follicular dendritic cells, BP3(+) stromal cells, and ERTR7(+) lymphoreticular cells. However, unlike the NALT of Ltalpha(-/-) mice, the NALT of Cxcl13(-/-) mice has peripheral node addressin(+) high endothelial venules (HEVs). In contrast, the NALT of plt/plt mice is nearly normal, with follicular dendritic cells, BP3(+) stromal cells, ERTR7(+) lymphoreticular cells, and peripheral node addressin(+) HEVs. Functionally, germinal center formation and switching to IgA are defective in the NALT of Ltalpha(-/-) and Cxcl13(-/-) mice. In contrast, CD8 T cell responses to influenza are impaired in Ltalpha(-/-) mice and plt/plt mice. Finally, the B and T cell defects in the NALT of Ltalpha(-/-) mice lead to delayed clearance of influenza from the nasal mucosa. Thus, the B and T cell defects in the NALT of Ltalpha(-/-) mice can be attributed to the impaired expression of CXCL13 and CCL19/CCL21, respectively, whereas impaired HEV development is directly due to the loss of LTalpha. PMID- 16210593 TI - V alpha 24-invariant NKT cells from patients with allergic asthma express CCR9 at high frequency and induce Th2 bias of CD3+ T cells upon CD226 engagement. AB - We have demonstrated that Valpha24(+)Vbeta11(+) invariant (Valpha24(+)i) NKT cells from patients with allergic asthma express CCR9 at high frequency. CCR9 ligand CCL25 induces chemotaxis of asthmatic Valpha24(+)i NKT cells but not the normal cells. A large number of CCR9-positive Valpha24(+)i NKT cells are found in asthmatic bronchi mucosa, where high levels of Th2 cytokines are detected. Asthmatic Valpha24(+)i NKT cells, themselves Th1 biased, induce CD3(+) T cells into an expression of Th2 cytokines (IL-4 and IL-13) in cell-cell contact manner in vitro. CD226 are overexpressed on asthmatic Valpha24(+)i NKT cells. CCL25/CCR9 ligation causes directly phosphorylation of CD226, indicating that CCL25/CCR9 signals can cross-talk with CD226 signals to activate Valpha24(+)i NKT cells. Prestimulation with immobilized CD226 mAb does not change ability of asthmatic Valpha24(+)i NKT cells to induce Th2-cytokine production, whereas soluble CD226 mAb or short hairpin RNA of CD226 inhibits Valpha24(+)i NKT cells to induce Th2 cytokine production by CD3(+) T cells, indicating that CD226 engagement is necessary for Valpha24(+)i NKT cells to induce Th2 bias of CD3(+) T cells. Our results are providing with direct evidence that aberration of CCR9 expression on asthmatic Valpha24(+)i NKT cells. CCL25 is first time shown promoting the recruitment of CCR9-expressing Valpha24(+)i NKT cells into the lung to promote other T cells to produce Th2 cytokines to establish and develop allergic asthma. Our findings provide evidence that abnormal asthmatic Valpha24(+)i NKT cells induce systemically and locally a Th2 bias in T cells that is at least partially critical for the pathogenesis of allergic asthma. PMID- 16210594 TI - Modulation of CD4 Th cell differentiation by ganglioside GD1a in vitro. AB - Cell surface gangliosides are shed by tumors into their microenvironment. In this study they inhibit cellular immune responses, including APC development and function, which is critical for Th1 and Th2 cell development. Using human dendritic cells (DCs) and naive CD4(+) T cells, we separately evaluated Th1 and Th2 development under the selective differentiating pressures of DC1-inducing pertussis toxin (PT) and DC2-inducing cholera toxin (CT). High DC IL-12 production after PT exposure and high DC IL-10 production after CT exposure were observed, as expected. However, when DCs were first preincubated with highly purified G(D1a) ganglioside, up-regulation of costimulatory molecules was blunted, and PT-induced IL-12 production was reduced, whereas CT-induced IL-10 production was increased. The combination of these effects could contribute to a block in the Th1 response. In fact, when untreated naive T cells were coincubated with ganglioside-preincubated, Ag-exposed DCs, naive Th cell differentiation into Th effector cells was reduced. Both the subsequent DC1-induced T cell production of IFN-gamma (Th1 marker) and DC2-induced T cell IL-4 production (Th2) were inhibited. Thus, ganglioside exposure of DC impairs, by at least two distinct mechanisms, the ability to induce Th differentiation, which could adversely affect the development of an effective cellular antitumor immune response. PMID- 16210595 TI - Conformational variation of surface class II MHC proteins during myeloid dendritic cell differentiation accompanies structural changes in lysosomal MIIC. AB - Dendritic cells (DC), uniquely among APC, express an open/empty conformation of MHC class II (MHC-II) proteins (correctly folded molecules lacking bound peptides). Generation and trafficking of empty HLA-DR during DC differentiation are investigated here. HLA-DR did not fold as an empty molecule in the endoplasmic reticulum/trans-Golgi network, did not derived from MHC/Ii complexes trafficking to the cell surface, but was generated after invariant chain degradation within lysosomal-like MHC-II rich compartments (MIIC). In pre-DC, generated from monocytes cultured in the presence of GM-CSF, Lamp-1(+)MHC-II(+) compartments are predominantly electron dense and, in these cells, empty MHC-II molecules accounts for as much as 20% of total surface HLA-DR. In immature DC, generated in presence of GM-CSF and IL-4, empty HLA-DR reside in multilamellar MIIC, but are scarcely observed at the cell surface. Thus, the morphology/composition of lysosomal MIIC at different DC maturational stages appear important for surface egression or intracellular retention of empty HLA DR. Ag loading can be achieved for the fraction of empty HLA-DR present in the "peptide-receptive" form. Finally, in vivo, APC-expressing surface empty HLA-DR were found in T cell areas of secondary lymphoid organs. PMID- 16210596 TI - Proinflammatory stimulation and pioglitazone treatment regulate peroxisome proliferator-activated receptor gamma levels in peripheral blood mononuclear cells from healthy controls and multiple sclerosis patients. AB - The peroxisome proliferator-activated receptor gamma (PPAR-gamma) belongs to a receptor superfamily of ligand-activated transcription factors involved in the regulation of metabolism and inflammation. Oral administration of PPAR-gamma agonists ameliorates the clinical course and histopathological features in experimental autoimmune encephalomyelitis, an animal model for multiple sclerosis (MS), and PPAR-gamma agonist treatment of PBMCs from MS patients suppresses PHA induced cell proliferation and cytokine secretion. These effects are pronounced when cells are preincubated with the PPAR-gamma agonists and reexposed at the time of stimulation, indicating a sensitizing effect. To characterize the mechanisms underlying this sensitizing effect, we analyzed PPAR-gamma expression in PMBCs of MS patients and healthy controls. Surprisingly, MS patients exhibited decreased PPAR-gamma levels compared with controls. PHA stimulation of PBMCs from healthy controls resulted in a significant loss of PPAR-gamma, which was prevented by in vitro preincubation of the cells or in vivo by long-term oral medication with the PPAR-gamma agonist pioglitazone. Differences in PPAR-gamma expression were accompanied by changes in PPAR-gamma DNA-binding activity, as preincubation with pioglitazone increased DNA binding of PPAR-gamma. Additionally, preincubation decreased NF-kappaB DNA-binding activity to control levels, whereas the inhibitory protein IkappaBalpha was increased. In MS patients, pioglitazone-induced increase in PPAR-gamma DNA-binding activity and decrease in NF-kappaB DNA-binding activity was only observed in the absence of an acute MS relapse. These results suggest that the sensitizing effect observed in the preincubation experiments is mediated by prevention of inflammation-induced suppression of PPAR-gamma expression with consecutive increase in PPAR-gamma DNA binding activity. PMID- 16210597 TI - Prevention of allergen-specific, Th2-biased immune responses in vivo: role of increased IL-12 and IL-18 responsiveness. AB - The factors that control development of adaptive responses to exogenous Ag remain incompletely understood. An ability to selectively direct immunity toward a specific phenotype would be of clinical benefit in numerous immunological disorders. Administration of chemically modified allergen glutaraldehyde polymerized OVA (OA-POL) leads to >90% reductions in murine IgE and >500-fold increases in IgG2c responses that develop upon subsequent immunization with native Ag. In the present study, we examine the mechanisms underlying this reorientation of the type 2 dominant response that would normally develop. Lack of endogenous IL-12 or IFN-gamma results in markedly reduced induction of IgG2c responses following OA-POL treatment, but only IFN-gamma(-/-) mice demonstrate reduced capacity to prevent IgE induction. This indicates that while both IL-12 and IFN-gamma are critical promoters of type 1 immunity, only IFN-gamma is required to maximally inhibit development of type 2 immune responses. Compared with OVA-immunized mice, CD69(+) T cells from OA-POL-immunized mice demonstrate elevated IL-12Rbeta(2), IL-18Ralpha, and IL-18Rbeta mRNA levels, as well as increased IFN-gamma production in response to rIL-12 or rIL-18 stimulation. Collectively, these data indicate that preventing induction of type 2 immune responses is critically dependent on altered T cell responsiveness to these cytokines. The finding that targeted, Ag-specific manipulation of IL-12 and IL-18 responsiveness can be used to shape the phenotype of the dominant immune response that develops suggests that specifically targeting IL-12 and IL-18 receptor expression may offer clinical options for clinical prophylaxis or intervention. PMID- 16210598 TI - Early presence of regulatory cells in transplanted rats rendered tolerant by donor-specific blood transfusion. AB - Mechanisms by which donor-specific blood transfusion (DSBT) promotes organ allograft acceptance are unclear. In a rat fully mismatched cardiac allograft model, we found that DSBT alone (without immunotherapy) induces the development of regulatory T cells (DSBT-Tregs) posttransplant, thereby shedding new light in the mechanisms of the transfusion effect. Compartments and timing of expansion, requirements, and phenotype of DSBT-Tregs are unknown. It is generally assumed that some time is necessary before Tregs develop. However, we show-by adoptive transfer from DSBT-tolerant into naive recipients: 1) the presence of DSBT-Tregs at 5 days posttransplant in spleen and lymph nodes; 2) their gradual expansion in these compartments; and 3) their presence in the graft 14 of 30 days posttransplant. DSBT-Tregs are donor specific and do not protect third-party allografts. Splenocytes from DSBT-treated nontransplanted recipients or from transplanted DSBT-untreated (rejecting) recipients do not transfer tolerance, indicating that both DSBT and graft are required for sufficient numbers of DSBT Tregs to develop. Thymectomy (or splenectomy) before DSBT (not at transplantation) abrogate DSBT-Tregs generation and tolerance, showing that thymus (and spleen) are required for DSBT-Tregs generation (not for expansion/maintenance). In contrast with other Tregs models, DSBT-Tregs activity is not restricted to CD4(+)CD25(+) but to CD4(+)CD45RC(-) cells, whereas CD4(+)CD45RC(+) cells act as effector cells and accelerate rejection. In conclusion, DSBT alone induces-rapidly posttransplant-the development of alloantigen-specific Tregs in lymphoid tissues and in the graft. DSBT, graft, thymus, and spleen are required for DSBT-Tregs generation. DSBT-Tregs in this model are CD4(+)CD45RC(-) (identical to Tregs protecting from autoimmunity in rats). PMID- 16210599 TI - Severe impairment of dendritic cell allostimulatory activity by Sendai virus vectors is overcome by matrix protein gene deletion. AB - Delivery of Ags to dendritic cells (DCs) plays a pivotal role in the induction of efficient immune responses ranging from immunity to tolerance. The observation that certain viral pathogens are able to infect DCs has led to a concept in which applications of recombinant viruses are used for Ag delivery with the potential benefit of inducing potent Ag-specific T cell responses directed against multiple epitopes. As a prerequisite for such an application, the infection of DCs by recombinant viruses should not interfere with their stimulatory capacity. In this context, we could show that an emerging negative-strand RNA viral vector system based on the Sendai virus (SeV) is able to efficiently infect monocyte-derived human DCs (moDCs). However, after infection with SeV wild type, both the response of DCs to bacterial LPS as a powerful mediator of DC maturation and the allostimulatory activity were severely impaired. Interestingly, using various recombinant SeV vectors that were devoid of single viral genes, we were able to identify the SeV matrix (M) protein as a key component in moDC functional impairment after viral infection. Consequently, use of M-deficient SeV vectors preserved the allostimulatory activity in infected moDCs despite an efficient expression of all other virally encoded genes, thereby identifying M-deficient vectors as a highly potent tool for the genetic manipulation of DCs. PMID- 16210600 TI - Mechanisms of hypotonicity-induced calcium signaling and integrin activation by arachidonic acid-derived inflammatory mediators in B cells. AB - We previously characterized the initial steps in the activation of novel (calcium permeant) nonselective cation channels (NSCCs) and calcium release-activated calcium channels in primary murine B lymphocytes. Phospholipase C products, namely diacylglycerol and d-myo-inositol 1,4,5-trisphosphate, were identified as proximal intracellular agonists of these respective channels following mechanical stimulation of B cells. However, neither the distal steps in NSCC activation nor the contribution of these channels to sustained mechanical signaling were defined in these previous studies. In this study, single cell measurements of intracellular Ca(2+) were used to define the mechanisms of NSCC activation and demonstrate a requirement for arachidonic acid liberated from diacylglycerol. Several arachidonic acid-derived derivatives were identified that trigger Ca(2+) entry into B cells, including the lipoxygenase product 5 hydroperoxyeicosatetranenoic acid and the cytochrome P450 hydroxylase product 20 hydroxyeicosatetraenoic; however, the cytochrome P450 epoxygenase product 5,6 epoxyeicosatrienoic acid is primarily responsible for hypotonicity-induced responses. In addition to regulating calcium entry, our data suggest that eicosanoid-activated NSCCs have a separate and direct role in regulating the avidity of integrins on B cells for extracellular matrix proteins, including ICAM 1 and VCAM-1. Thus, in addition to defining a novel osmotically activated signal transduction pathway in B cells, our results have broad implications for understanding how inflammatory mediators dynamically and rapidly regulate B cell adhesion and trafficking. PMID- 16210601 TI - Conjugated linoleic acid suppresses NF-kappa B activation and IL-12 production in dendritic cells through ERK-mediated IL-10 induction. AB - Polyunsaturated fatty acids (PUFA) have been shown to modulate immune responses and have therapeutic effects in inflammatory disorders. However, the influence of PUFA on dendritic cells (DC), key cells of the innate immune system in shaping adaptive immune responses, has not yet been defined. In this study, we examine the effects of the cis-9, trans-11 isomer of conjugated linoleic acid (c9, t11 CLA), a dietary PUFA found in meat and dairy products, on murine DC activation. Treatment of DC with c9, t11-CLA suppressed LPS-induced IL-12, enhanced IL-10R expression, and enhanced IL-10 production at the transcriptional and protein level. The suppression of IL-12 by c9, t11-CLA was found to be IL-10 dependent. We investigated the involvement of the MAPK, ERK, and the transcription factor, NF-kappaB, in this IL-10-mediated effect. c9, t11-CLA enhanced ERK activation after LPS stimulation, and inhibition of ERK resulted in abrogation of IL-10 and recovery of IL-12 production. c9, t11-CLA decreased NF-kappaB:DNA binding after LPS stimulation, which was concomitant with delayed translocation of NF-kappaBp65 into the nucleus and an increase in IkappaBalpha. These effects were reversed by addition of a neutralizing anti-IL-10 Ab. Our findings demonstrate that c9, t11 CLA suppresses IL-12 production by LPS-stimulated DC by ERK mediated IL-10 induction. Furthermore, these IL-10-mediated effects are dependent on inhibition of NF-kappaB activation. This is the first study to demonstrate that c9, t11-CLA can enhance transcription and production of the anti-inflammatory cytokine IL-10, while inhibiting the Th1-promoting cytokine IL-12, and may explain certain of its immunosuppressive properties. PMID- 16210602 TI - Chronic morphine treatment promotes specific Th2 cytokine production by murine T cells in vitro via a Fas/Fas ligand-dependent mechanism. AB - Improper homeostasis of Th1 and Th2 cell differentiation can promote pathological immune responses such as autoimmunity and asthma. A number of factors govern the development of these cells including TCR ligation, costimulation, death effector expression, and activation-induced cell death (AICD). Although chronic morphine administration has been shown to selectively promote Th2 development in unpurified T cell populations, the direct effects of chronic morphine on Th cell skewing and cytokine production by CD4(+) T cells have not been elucidated. We previously showed that morphine enhances Fas death receptor expression in a T cell hybridoma and human PBL. In addition, we have demonstrated a role for Fas, Fas ligand (FasL), and TRAIL in promoting Th2 development via killing of Th1 cells. Therefore, we analyzed whether the ability of morphine to affect Th2 cytokine production was mediated by regulation of Fas, FasL, and TRAIL expression and AICD directly in purified Th cells. We found that morphine significantly promoted IL-4 and IL-13 production but did not alter IL-5 or IFN-gamma. Furthermore, morphine enhanced the mRNA expression of Fas, FasL and TRAIL and promoted Fas-mediated AICD of CD4(+) T cells. Additionally, blockade of Fas/FasL interaction by anti-FasL inhibited the morphine-induced production of IL-4 and IL 13 and AICD of CD4(+) T cells. These results suggest that morphine preferentially enhances Th2 cell differentiation via killing of Th1 cells in a Fas/FasL dependent manner. PMID- 16210603 TI - Killer cell Ig-like receptor-dependent signaling by Ig-like transcript 2 (ILT2/CD85j/LILRB1/LIR-1). AB - Inhibitory killer cell Ig-like receptors (KIR) signal by recruitment of the tyrosine phosphatase Src homology region 2 domain-containing phosphatase-1 to ITIM. In the present study, we show that, surprisingly, KIR lacking ITIM are able to signal and inhibit in the human NK cell line NK92, but not in mouse NK cells. Signaling by mutant KIR is weaker than the wild-type receptor, does not require the transmembrane or cytoplasmic tail of KIR, and is blocked by overexpression of a catalytically inactive Src homology region 2 domain-containing phosphatase-1 molecule. We also demonstrate that mutant KIR signaling is blocked by Abs, which disrupt the interaction between KIR and human leukocyte Ag-C or Abs, which block the interaction between Ig-like transcript 2 (ILT2) and the alpha3 domain of HLA class I molecules. Thus, although ILT2 expressed in NK92 is insufficient to signal in response to human leukocyte Ag-C alone, ILT2 can signal in a KIR dependent manner revealing functional cooperation between receptors encoded by two distinct inhibitory receptor families. PMID- 16210604 TI - Heterogeneity of Flt3-expressing multipotent progenitors in mouse bone marrow. AB - Mechanisms of lymphoid and myeloid lineage choice by hemopoietic stem cells remain unclear. In this study we show that the multipotent progenitor (MPP) population, which is immediately downstream of hemopoietic stem cells, is heterogeneous and can be subdivided in terms of VCAM-1 expression. VCAM-1(+) MPPs were fully capable of differentiating into both lymphoid and myeloid lineages. In contrast, VCAM-1(-) MPPs gave rise to lymphocytes predominately in vivo. T and B cell development from VCAM-1(-) MPPs was 1 wk faster than that from VCAM-1(+) MPPs. Furthermore, VCAM-1(+) MPPs gave rise to common myeloid progenitors and VCAM-1(-) MPPs in vivo, indicating that VCAM-1(-) MPPs are progenies of VCAM-1(+) MPPs. VCAM-1(-) MPPs, in turn, developed into lymphoid lineage-restricted common lymphoid progenitors. These results establish a hierarchy of developmental relationship between MPP subsets and lymphoid and myeloid progenitors. In addition, VCAM-1(+) MPPs may represent the branching point between the lymphoid and myeloid lineages. PMID- 16210605 TI - Paradoxical anti-inflammatory actions of TNF-alpha: inhibition of IL-12 and IL-23 via TNF receptor 1 in macrophages and dendritic cells. AB - IL-12 and TNF-alpha are central proinflammatory cytokines produced by macrophages and dendritic cells. Disregulation of TNF-alpha is associated with sepsis and autoimmune diseases such as rheumatoid arthritis. However, new evidence suggests an anti-inflammatory role for TNF-alpha. TNF-alpha-treated murine macrophages produced less IL-12p70 and IL-23, after stimulation with IFN-gamma and LPS. Frequency of IL-12p40-producing macrophages correspondingly decreased as measured by intracellular cytokine staining. IL-12p40 production was also inhibited in dendritic cells. TNFR1 was established as the main receptor involved in IL-12p40 regulation, because IL-12p40 levels were not affected by TNF-alpha in TNFR1(-/-) derived macrophages. Macrophages activated during Listeria monocytogenes infection were more susceptible to inhibition by TNF-alpha than cells from naive animals, which suggests a regulatory role for TNF-alpha in later stages of infection. This nonapoptotic anti-inflammatory regulation of IL-12 and IL-23 is an important addition to the multitude of TNF-alpha-induced responses determined by cell-specific receptor signaling. PMID- 16210606 TI - Tight regulation of IFN-gamma transcription and secretion in immature and mature B cells by the inhibitory MHC class I receptor, Ly49G2. AB - To complete their maturation and to participate in the humoral immune response, immature B cells that leave the bone marrow are targeted to specific areas in the spleen, where they differentiate into mature cells. Previously, we showed that immature B cells actively down-regulate their integrin-mediated migration to lymph nodes or sites of inflammation, enabling their targeting to the spleen to allow their final maturation. This inhibition is mediated by IFN-gamma, which is transcribed and secreted at low levels by these immature B cells and is down regulated at the mature stage. The activating MHC class I receptor, Ly49D, which is expressed at high levels on immature B cells, stimulates this IFN-gamma secretion. In this study we show that B cells coexpress the inhibitory MHC class I receptor, Ly49G2. In addition, we demonstrate a tight regulation in the expression of the Ly49 family members on B cells that depends on their cell surface levels. High levels of Ly49G2 have a dominant inhibitory effect on Ly49D expressed at low levels on immature bone marrow and mature B cells, resulting in inhibition of IFN-gamma secretion. However, low levels of the inhibitory receptor, Ly49G2, coexpressed with high levels of the activating receptor, Ly49D, on the immigrating immature B cells enable the secretion of specific low levels of IFN-gamma. This expression pattern insures the inhibitory control of peripheral immature B cell to prevent premature encounter with an Ag while enabling entry to the lymph nodes during the mature stage. PMID- 16210607 TI - Rapid production of TNF-alpha following TCR engagement of naive CD8 T cells. AB - The acquisition of effector functions by naive CD8 T cells following TCR engagement is thought to occur sequentially with full functionality being gained only after the initiation of division. We show that naive CD8 T cells are capable of immediate effector function following TCR engagement, which stimulates the rapid production of TNF-alpha. Stimulation of splenocytes from naive mice of differing genetic backgrounds with anti-CD3epsilon mAb resulted in significant production of TNF-alpha by naive CD8 T cells within 5 h. Moreover, naive lymphocytic choriomeningitis virus-specific TCR-transgenic CD8 T cells stimulated with either their cognate peptide ligand or virus-infected cells produced TNF alpha as early as 2 h poststimulation, with production peaking by 4 h. Naive CD8 T cells produced both membrane-bound and soluble TNF-alpha. Interfering with TNF alpha activity during the initial encounter between naive CD8 T cells and Ag loaded dendritic cells altered the maturation profile of the APC and diminished the overall viability of the APC population. These findings suggest that production of TNF-alpha by naive CD8 T cells immediately after TCR engagement may have an unappreciated impact within the local environment where Ag presentation is occurring and potentially influence the development of immune responses. PMID- 16210608 TI - Estrogen enhances susceptibility to experimental autoimmune myasthenia gravis by promoting type 1-polarized immune responses. AB - Myasthenia gravis (MG) is an organ-specific autoimmune disease caused in most cases by autoantibodies against the nicotinic acetylcholine receptor (AChR). It is now well documented that many autoimmune diseases, including MG, are more prevalent in women than in men, and that fluctuations in disease severity occur during pregnancy. These observations raise the question of the potential role of sex hormones, such as estrogens, as mediators of sex differences in autoimmunity. In the present study, we have analyzed the effect of 17beta-estradiol (E2) on the pathogenesis of experimental autoimmune myasthenia gravis (EAMG), an animal model of MG. We show that treatment with E2 before Ag priming is necessary and sufficient to promote AChR-specific Th1 cell expansion in vivo. This time-limited exposure to E2 enhances the production of anti-AChR IgG2a(b) (specific for b allotype; e.g., B6) and IgG2b, but not IgG1, and significantly increases the severity of EAMG in mice. Interestingly, the E2-mediated augmentation in AChR specific Th1 response correlates with an enhanced production of IL-12 by splenic APCs through the recruitment of CD8alpha(+) dendritic cells. These data provide the first evidence that estrogen enhances EAMG, and sheds some light on the role of sex hormones in immune responses and susceptibility to autoimmune disease in women. PMID- 16210609 TI - Both regulatory T cells and antitumor effector T cells are primed in the same draining lymph nodes during tumor progression. AB - The peripheral tolerance mechanism prevents effective antitumor immunity, even though tumor cells possess recognizable tumor-associated Ags. Recently, it has been elucidated that regulatory T cells (Treg) play a critical role in maintaining not only self-tolerance, but also tolerance of tumor cells. However, because the Treg that maintain self-tolerance arise naturally in the thymus and are thought to be anergic in peripheral, it is still unclear where and when Treg for tumor cells are generated. In this study we analyze tumor-draining lymph nodes (LNs) and demonstrate that both antitumor effector T cells and Treg capable of abrogating the antitumor reactivity of the effector T cells are primed in the same LNs during tumor progression. The regulatory activity generated in tumor draining LNs exclusively belonged to the CD4(+) T cell subpopulation that expresses both CD25 and a high level of CD62L. Forkhead/winged helix transcription factor gene expression was detected only in the CD62L(high)CD4(+)CD25(+) T cells. CD62L(high)CD4(+)CD25(+) Treg and CD62L(low)CD4(+)CD25(+) T cells, which possess effector T cell functions, had comparable expression of LFA-1, VLA-4, CTLA-4, lymphocyte activation gene-3, and glucocorticoid-induced TNFR. Thus, only CD62L expression could distinguish regulatory CD4(+)CD25(+) cells from effector CD4(+)CD25(+) cells in draining LNs as a surface marker. The Treg generated in tumor-draining LNs possess the same functional properties as the Treg that arise naturally in the thymus but recognize tumor-associated Ag. CD62L(high)CD4(+)CD25(+) Treg contained a subpopulation that expressed CD86. Blocking experiments revealed that ligation of CTLA-4 on effector T cells by CD86 on Treg plays a pivotal role in regulating CD4(+) effector T cells. PMID- 16210610 TI - Receptor editing can lead to allelic inclusion and development of B cells that retain antibodies reacting with high avidity autoantigens. AB - Receptor editing is a major B cell tolerance mechanism that operates by secondary Ig gene rearrangements to change the specificity of autoreactive developing B cells. In the 3-83Igi mouse model, receptor editing operates in every autoreactive anti-H-2K(b) B cell, providing a novel receptor without additional cell loss. Despite the efficiency of receptor editing in generating nonautoreactive Ag receptors, we show in this study that this process does not inactivate the autoantibody-encoding gene(s) in every autoreactive B cell. In fact, receptor editing can generate allelically and isotypically included B cells that simultaneously express the original autoreactive and a novel nonautoreactive Ag receptors. Such dual Ab-expressing B cells differentiate into transitional and mature B cells retaining the expression of the autoantibody despite the high avidity interaction between the autoantibody and the self-Ag in this system. Moreover, we find that these high avidity autoreactive B cells retain the autoreactive Ag receptor within the cell as a consequence of autoantigen engagement and through a Src family kinase-dependent process. Finally, anti-H 2K(b) IgM autoantibodies are found in the sera of older 3-83Igi mice, indicating that dual Ab-expressing autoreactive B cells are potentially functional and capable of differentiating into IgM autoantibody-secreting plasma cells under certain circumstances. These results demonstrate that autoreactive B cells reacting with ubiquitous membrane bound autoantigens can bypass mechanisms of central tolerance by coexpressing nonautoreactive Abs. These dual Ab-expressing autoreactive B cells conceal their autoantibodies within the cell manifesting a superficially tolerant phenotype that can be partially overcome to secrete IgM autoantibodies. PMID- 16210611 TI - Treatment of mice with the suppressor of cytokine signaling-1 mimetic peptide, tyrosine kinase inhibitor peptide, prevents development of the acute form of experimental allergic encephalomyelitis and induces stable remission in the chronic relapsing/remitting form. AB - We have previously characterized a novel tyrosine kinase inhibitor peptide (Tkip) that is a mimetic of suppressor of cytokine signaling 1 (SOCS-1) and inhibits JAK2 phosphorylation of the transcription factor STAT1alpha. We show in this study that Tkip protects mice against experimental allergic encephalomyelitis (EAE), an animal model for multiple sclerosis. Mice are immunized with myelin basic protein (MBP) for induction of disease. Tkip (63 mug) administered every other day suppressed the development of acute EAE in 75% of New Zealand White (NZW) mice. Furthermore, Tkip completely protected SJL/J mice, which where induced to get the relapsing/remitting form of EAE, against relapses compared with control groups in which >70% of the mice relapsed after primary incidence of disease. Protection of mice by Tkip was similar to that seen with the type I IFN, IFN-tau. Protection of mice correlated with lower MBP Ab titers in Tkip-treated groups as well as suppression of MBP-induced proliferation of splenocytes taken from EAE-afflicted mice. Cessation of Tkip and IFN-tau administration resulted in SJL/J mice relapsing back into disease. Prolonged treatment of mice with Tkip produced no evidence of cellular toxicity or weight loss. Consistent with its JAK2 inhibitory function, Tkip also inhibited the activity of the inflammatory cytokine TNF-alpha, which uses the STAT1alpha transcription factor. The data presented in this study show that Tkip, like the type I IFN, IFN-tau, inhibits both the autoreactive cellular and humoral responses in EAE and ameliorates both the acute and chronic relapsing/remitting forms of EAE. PMID- 16210612 TI - Human invariant NKT cells are required for effective in vitro alloresponses. AB - NKT cells are a small subset of regulatory T cells conserved in humans and mice. In humans they express the Valpha24Jalpha18 invariant chain (hence invariant NKT (iNKT) cells) and are restricted by the glycolipid-presenting molecule CD1d. In mice, iNKT cells may enhance or inhibit anti-infectious and antitumor T cell responses but suppress autoimmune and alloreactive responses. We postulated that iNKT cells might also modulate human alloreactive responses. Using MLR assays we demonstrate that in the presence of the CD1d-presented glycolipid alpha galactosylceramide (alphaGC) alloreactivity is enhanced (37 +/- 12%; p < 0.001) in an iNKT cell-dependent manner. iNKT cells are activated early during the course of the MLR, presumably by natural ligands. In MLR performed without exogenous ligands, depletion of iNKT cells significantly diminished the alloresponse in terms of proliferation (58.8 +/- 24%; p < 0.001) and IFN-gamma secretion (43.2 +/- 15.2%; p < 0.001). Importantly, adding back fresh iNKT cells restored the reactivity of iNKT cell-depleted MLR to near baseline levels. CD1d blocking mAbs equally reduced the reactivity of the iNKT cell-replete and depleted MLR compared with IgG control, indicating that the effect of iNKT cells in the in vitro alloresponse is CD1d-dependent. These findings suggest that human iNKT cells, although not essential for its development, can enhance the alloreactive response. PMID- 16210613 TI - Human embryonic stem cell-derived NK cells acquire functional receptors and cytolytic activity. AB - Human embryonic stem cells (hESCs) provide a unique resource to analyze early stages of human hematopoiesis. However, little is known about the ability to use hESCs to evaluate lymphocyte development. In the present study, we use a two-step culture method to demonstrate efficient generation of functional NK cells from hESCs. The CD56(+)CD45(+) hESC-derived lymphocytes express inhibitory and activating receptors typical of mature NK cells, including killer cell Ig-like receptors, natural cytotoxicity receptors, and CD16. Limiting dilution analysis suggests that these cells can be produced from hESC-derived hemopoietic progenitors at a clonal frequency similar to CD34(+) cells isolated from cord blood. The hESC-derived NK cells acquire the ability to lyse human tumor cells by both direct cell-mediated cytotoxicity and Ab-dependent cellular cytotoxicity. Additionally, activated hESC-derived NK cells up-regulate cytokine production. hESC-derived lymphoid progenitors provide a novel means to characterize specific cellular and molecular mechanisms that lead to development of specific human lymphocyte populations. These cells may also provide a source for innovative cellular immune therapies. PMID- 16210614 TI - Critical role for the tapasin-docking site of TAP2 in the functional integrity of the MHC class I-peptide-loading complex. AB - The transporter associated with Ag processing (TAP) translocates antigenic peptides into the endoplasmic reticulum for binding onto MHC class I (MHC I) molecules. Tapasin organizes a peptide-loading complex (PLC) by recruiting MHC I and accessory chaperones to the N-terminal regions (N domains) of the TAP subunits TAP1 and TAP2. To investigate the function of the tapasin-docking sites of TAP in MHC I processing, we expressed N-terminally truncated variants of TAP1 and TAP2 in combination with wild-type chains, as fusion proteins or as single subunits. Strikingly, TAP variants lacking the N domain in TAP2, but not in TAP1, build PLCs that fail to generate stable MHC I-peptide complexes. This correlates with a substantially reduced recruitment of accessory chaperones into the PLC demonstrating their important role in the quality control of MHC I loading. However, stable surface expression of MHC I can be rescued in post-endoplasmic reticulum compartments by a proprotein convertase-dependent mechanism. PMID- 16210615 TI - A CXCR4-dependent chemorepellent signal contributes to the emigration of mature single-positive CD4 cells from the fetal thymus. AB - Developing thymocytes undergo maturation while migrating through the thymus and ultimately emigrate from the organ to populate peripheral lymphoid tissues. The process of thymic emigration is controlled in part via receptor-ligand interactions between the chemokine stromal-derived factor (SDF)-1, and its cognate receptor CXCR4, and sphingosine 1-phosphate (S1P) and its receptor S1PR. The precise mechanism by which S1P/S1PR and CXCR4/SDF-1 contribute to thymic emigration remains unclear. We proposed that S1P-dependent and -independent mechanisms might coexist and involve both S1P-induced chemoattraction and SDF-1 mediated chemorepulsion or fugetaxis of mature thymocytes. We examined thymocyte emigration in thymi from CXCR4-deficient C57BL/6 embryos in a modified assay, which allows the collection of CD62L(high) and CD69(low) recent thymic emigrants. We demonstrated that single-positive (SP) CD4 thymocytes, with the characteristics of recent thymic emigrants, failed to move away from CXCR4 deficient fetal thymus in vitro. We found that the defect in SP CD4 cell emigration that occurred in the absence of CXCR4 signaling was only partially overcome by the addition of the extrathymic chemoattractant S1P and was not associated with abnormalities in thymocyte maturation and proliferative capacity or integrin expression. Blockade of the CXCR4 receptor in normal thymocytes by AMD3100 led to the retention of mature T cells in the thymus in vitro and in vivo. The addition of extrathymic SDF-1 inhibited emigration of wild-type SP cells out of the thymus by nullifying the chemokine gradient. SDF-1 was also shown to elicit a CXCR4-dependent chemorepellent response from fetal SP thymocytes. These novel findings support the thesis that the CXCR4-mediated chemorepellent activity of intrathymic SDF-1 contributes to SP thymocyte egress from the fetal thymus. PMID- 16210616 TI - Protein kinase C-theta is an early survival factor required for differentiation of effector CD8+ T cells. AB - CD8(+) T cells are crucial for host defense against invading pathogens and malignancies. However, relatively little is known about intracellular signaling events that control the genetic program of their activation and differentiation. Using CD8(+) T cells from TCR-transgenic mice crossed to protein kinase C-theta (PKCtheta)-deficient mice, we report that PKCtheta is not required for Ag-induced CD8(+) T cell proliferation, but is important for T cell survival and differentiation into functional, cytokine-producing CTLs. Ag-stimulated PKCtheta( /-) T cells underwent accelerated apoptosis associated with deregulated expression of Bcl-2 family proteins and displayed reduced activation of ERKs and JNKs. Some defects in the function of PKCtheta(-/-) T cells (poor survival and reduced Bcl-2 and Bcl-x(L) expression, CTL activity, and IFN-gamma expression) were partially or fully restored by coculture with wild-type T cells or by addition of exogenous IL-2, whereas others (increased Bim(EL) expression and TNF alpha production) were not. These findings indicate that PKCtheta, although not essential for initial Ag-induced proliferation, nevertheless plays an important role in promoting and extending T cell survival, thereby enabling the complete genetic program of effector CD8(+) differentiation. The requirement for PKCtheta in different types of T cell-dependent responses may, therefore, depend on the overall strength of signaling by the TCR and costimulatory receptors and may reflect, in addition to its previously established role in activation, an important, hitherto unappreciated, role in T cell survival. PMID- 16210617 TI - Attenuation of Th1 response in decoy receptor 3 transgenic mice. AB - The soluble decoy receptor 3 (DcR3) is a member of the TNFR superfamily. Because DcR3 is up-regulated in tumor tissues and is detectable in the sera of cancer patients, it is regarded as an immunosuppressor to down-regulate immune responses. To understand the function of DcR3 in vivo, we generated transgenic mice overexpressing DcR3 systemically. In comparison with HNT-TCR (HNT) transgenic mice, up-regulation of IL-4 and IL-10 and down-regulation of IFN gamma, IL-12, and TNF-alpha were observed in the influenza hemagglutinin(126-138) peptide-stimulated splenocytes of HNT-DcR3 double-transgenic mice. When infected with Listeria monocytogenes, DcR3 transgenic mice show attenuated expression of IFN-gamma as well as increased susceptibility to infection. The Th2 cell-biased phenotype in DcR3 transgenic mice is attributed to decreased IL-2 secretion by T cells, resulting in the suppression of IL-2 dependent CD4(+) T cell proliferation. This suggests that DcR3 might help tumor growth by attenuating the Th1 response and suppressing cell-mediated immunity. PMID- 16210618 TI - Estrogen selectively promotes the differentiation of dendritic cells with characteristics of Langerhans cells. AB - The steroid hormone estrogen regulates the differentiation, survival, or function of diverse immune cells. Previously, we found that physiological amounts of 17beta-estradiol act via estrogen receptors (ER) to promote the GM-CSF-mediated differentiation of dendritic cells (DC) from murine bone marrow progenitors in ex vivo cultures. Of the two major subsets of CD11c(+) DC that develop in these cultures, estrogen is preferentially required for the differentiation of a CD11b(int)Ly6C(-) population, although it also promotes increased numbers of a CD11b(high)Ly6C(+) population. Although both DC subsets express ERalpha, only the CD11b(high)Ly6C(+) DC express ERbeta, perhaps providing a foundation for the differential regulation of these two DC types by estrogen. The two DC populations exhibit distinct phenotypes in terms of capacity for costimulatory molecule and MHC expression, and Ag internalization, which predict functional differences. The CD11b(int)Ly6C(-) population shows the greatest increase in MHC and CD86 expression after LPS activation. Most notably, the estrogen-dependent CD11b(int)Ly6C(-) DC express langerin (CD207) and contain Birbeck granules characteristic of Langerhans cells. These data show that estrogen promotes a DC population with the unique features of epidermal Langerhans cells and suggest that differentiation of Langerhans cells in vivo will be dependent upon local estrogen levels and ER-mediated signaling events in skin. PMID- 16210619 TI - Functional characterization of human cysteinyl leukotriene 1 receptor gene structure. AB - The 5-lipoxygenase pathway has been strongly implicated in the pathogenesis of chronic inflammatory disorders, such as bronchial asthma and atherosclerosis. Cysteinyl leukotrienes (cysLTs), 5-lipoxygenase pathway products, are recognized now not only as important factors in asthmatic inflammation, but also as mediators of cell trafficking and innate immune responses. To study a role of cysLTs in inflammatory reactions we have characterized the gene structure of human cysteinyl leukotriene receptor type I (cysLT(1)R). The cysLT(1)R gene consists of 5 exons that are variably spliced and a single promoter region with multiple transcription start sites. Four different cysLT(1)R transcripts were identified. RT-PCR showed dominant and wide expression of the transcript I, containing exons 1, 4, and 5, with the strongest presence in blood leukocytes, spleen, thymus, lung, and heart. The expression of cysLT(1)R is functionally regulated at the transcriptional level by IL-4 through a STAT6 response element localized to the proximal cysLT(1)R promoter region. IL-4 stimulation increased cysLT(1)R mRNA (real-time PCR) and surface protein expression (flow cytometry) in a time-dependent fashion. CysLTs (LTD(4) and LTC(4)) induced an increased production of a potent monocyte chemoattractant CCL2 (MCP-1) in IL-4-primed THP-1 cells in a dose-dependent manner. This effect was effectively inhibited by the cysLT(1)R-selective antagonist MK571 in a dose-dependent manner and only partially by a nonselective cysLT(1)R/cysLT(2)R inhibitor BAY-u9773, implying a cysLT(1)R-mediated mechanism. Thus, cysLTs signaling through cysLT(1)R might contribute to inflammatory reactions by cooperating with IL-4 in enhanced CCL2 production in human monocytic cells. PMID- 16210620 TI - Protein acetylation regulates both PU.1 transactivation and Ig kappa 3' enhancer activity. AB - Igkappa gene expression and chromatin structure change during B cell development. At the pre-B cell stage, the locus is relatively hypoacetylated on histone H3, whereas it is hyperacetylated at the plasma cell stage. We find in this study that the histone deacetylase inhibitor, trichostatin A (TSA) stimulated 3' enhancer activity through the PU.1 binding site. TSA also stimulated PU.1 transactivation potential. PU.1 activity was increased by the coactivator acetyltransferase protein, p300, and p300 physically interacted with PU.1 residues 7-30. PU.1 served as a substrate for p300 and was acetylated on lysine residues 170, 171, 206, and 208. Mutation of PU.1 lysines 170 and 171 did not affect PU.1 DNA binding, but did lower the ability of PU.1 to activate transcription in association with p300. Lysine 170 was acetylated in pre-B cells and plasmacytoma cells, but TSA treatment did not stimulate PU.1 acetylation at this residue arguing that a second mechanism can stimulate 3' enhancer activity. Using chromatin immunoprecipitation assays we found that TSA caused preferential acetylation of histone H3 at the 3' enhancer. The relevance of these studies for PU.1 function in transcription and hemopoietic development is discussed. PMID- 16210621 TI - Hypermutation at A-T base pairs: the A nucleotide replacement spectrum is affected by adjacent nucleotides and there is no reverse complementarity of sequences flanking mutated A and T nucleotides. AB - Hypermutation is thought to be a two-phase process. The first phase is via the action of activation-induced cytidine deaminase (AID), which deaminates C nucleotides in WRC motifs. This results in the RGYW/WRCY hot spot motifs for mutation from G and C observed in vivo. The resemblance between the hot spot for C mutations and the reverse complement of that for G mutations implies a process acting equally on both strands of DNA. The second phase of hypermutation generates mutations from A and T and exhibits strand bias, with more mutations from A than T. Although this does not concur with the idea of one mechanism acting equally on both strands, it has been suggested that the AT mutator also has a reversible motif; WA/TW. We show here that the motifs surrounding the different substitutions from A vary significantly; there is no single targeting motif for all A mutations. Sequence preferences associated with mutations from A more likely reflect an influence of adjacent nucleotides over what the A mutates "to." This influence tends toward "like" replacements: Purines (A or G) in the 5' position bias toward replacement by another purine (G), whereas replacement with pyrimidines (C or T) is more likely if the preceding base is also a pyrimidine. There is no reverse complementarity in these observations, in that similar influences of nucleotides adjacent to T are not seen. Hence, WA and TW should not be considered as reverse complement hot spot motifs for A and T mutations. PMID- 16210622 TI - Sequence motifs in IL-4R alpha mediating cell-cycle progression of primary lymphocytes. AB - IL-4 signaling through the IL-4Ralpha chain regulates the development and proliferation of the Th2 lineage of effector CD4(+) T cells. Analyses of the IL 4R in factor-dependent cell lines led to the development of two apparently conflicting models of the primary structural determinants of IL-4R-mediated proliferative signaling. In one model, proliferation was dependent on the first conserved tyrosine in the cytoplasmic tail (Y1), while in the second, proliferation was independent of cytoplasmic tyrosines. We found that in activated primary T cells, mutation of only the Y1 residue resulted in a modest decrease in IL-4-induced S phase entry, a further decrease in cell-cycle completion, and a complete failure of IL-4 to induce p70S6 kinase phosphorylation. Consistent with a role for the PI3K/mammalian target of rapamycin pathway in mediating cytokine acceleration of G(2)/M transit, pretreatment of activated T cells with rapamycin resulted in only a modest decrease in IL-4-induced S phase entry, but a total block of cell-cycle completion. Strikingly, IL-4Ralpha chains that lacked all cytoplasmic tyrosines were competent to signal for STAT5 phosphorylation, mediated efficient S phase entry, and promoted cell-cycle progression. The ability of tyrosine-deficient IL 4Rs to mediate proliferative signaling and STAT phosphorylation was absolutely dependent on the presence of an intact ID-1 region. These findings show that IL 4Ralpha lacking cytoplasmic tyrosine residues is competent to induce ID-1 dependent proliferation, and indicate that IL-4 can promote G(2)/M progression via activation of the mammalian target of rapamycin pathway initiated at the Y1 residue. PMID- 16210623 TI - Allele-specific regulation of TCR beta variable gene segment chromatin structure. AB - Allelic exclusion of the murine Tcrb locus is imposed at the level of recombination and restricts each cell to produce one functional VDJbeta rearrangement. Allelic exclusion is achieved through asynchronous Vbeta to DJbeta recombination as well as feedback inhibition that terminates recombination once a functional rearrangement has occurred. Because the accessibility of Vbeta gene segment chromatin is diminished as thymocytes undergo allelic exclusion at the CD4(-)CD8(-) (double-negative) to CD4(+)CD8(+) (double-positive) transition, chromatin regulation was thought to be an important component of the feedback inhibition process. However, previous studies of chromatin regulation addressed the status of Tcrb alleles using genetic models in which both alleles remained in a germline configuration. Under physiological conditions, developing thymocytes would undergo Vbeta to DJbeta recombination on one or both alleles before the enforcement of feedback. On rearranged alleles, Vbeta gene segments that in germline configuration are regulated independently of the Tcrb enhancer are now brought into its proximity. We show in this study that in contrast to Vbeta segments on a nonrearranged allele, those situated upstream of a functionally rearranged Vbeta segment are contained in active chromatin as judged by histone H3 acetylation, histone H3 lysine 4 (K4) methylation, and germline transcription. Nevertheless, these Vbeta gene segments remain refractory to recombination in double-positive thymocytes. These results suggest that a unique feedback mechanism may operate independent of chromatin structure to inhibit Vbeta to DJbeta recombination after the double-negative stage of thymocyte development. PMID- 16210624 TI - Splenic marginal zone dendritic cells mediate the cholera toxin adjuvant effect: dependence on the ADP-ribosyltransferase activity of the holotoxin. AB - The in vivo mechanisms of action of most vaccine adjuvants are poorly understood. In this study, we present data in mice that reveal a series of critical interactions between the cholera toxin (CT) adjuvant and the dendritic cells (DC) of the splenic marginal zone (MZ) that lead to effective priming of an immune response. For the first time, we have followed adjuvant targeting of MZ DC in vivo. We used CT-conjugated OVA and found that the Ag selectively accumulated in MZ DC following i.v. injections. The uptake of Ag into DC was GM1 ganglioside receptor dependent and mediated by the B subunit of CT (CTB). The targeted MZ DC were quite unique in their phenotype: CD11c(+), CD8alpha(-), CD11b(-), B220(-), and expressing intermediate or low levels of MHC class II and DEC205. Whereas CTB only delivered the Ag to MZ DC, the ADP-ribosyltransferase activity of CT was required for the maturation and migration of DC to the T cell zone, where these cells distinctly up-regulated CD86, but not CD80. This interaction appeared to instruct Ag-specific CD4(+) T cells to move into the B cell follicle and strongly support germinal center formations. These events may explain why CT-conjugated Ag is substantially more immunogenic than Ag admixed with soluble CT and why CTB conjugated Ag can tolerize immune responses when given orally or at other mucosal sites. PMID- 16210625 TI - Human high mobility group box transcription factor 1 affects thymocyte development and transgene variegation. AB - It has been shown previously that a human CD2 (hCD2) disabled locus control region (LCR) transgene is unable to establish an open chromatin configuration in all the T cells, and this leads to position effect variegation of the transgene. In this study we show that thymus-specific overexpression of human high mobility group box transcription factor 1 (HBP1), a transcription factor that binds a specific sequence within the hCD2 LCR, affects thymus cellularity as well as the number of CD8(+) thymocytes in two independent transgenic mouse lines and increases the proportion of T cells that fully activate the transgenic locus in hCD2 variegating mice in a sequence-specific dependent manner. This finding suggests that overexpression of HBP1 can affect lineage commitment and can relieve the suppressive influence of heterochromatin, allowing thymocytes to express the variegating target locus more efficiently. These effects could be the result of direct HBP1 action on LCR activity. Alternatively, the extra HBP1 molecules may sequester repressive elements away from the LCR, thus allowing transcription permissive states to form on the transgene locus. PMID- 16210626 TI - BMP signaling is required for normal thymus development. AB - The microenvironment of the thymus fosters the generation of a diverse and self tolerant T cell repertoire from a pool of essentially random specificities. Epithelial as well as mesenchymal cells contribute to the thymic stroma, but little is known about the factors that allow for communication between the two cells types that shape the thymic microenvironment. In this study, we investigated the role of bone morphogenetic protein (BMP) signaling in thymus development. Transgenic expression of the BMP antagonist Noggin in thymic epithelial cells under the control of a Foxn1 promoter in the mouse leads to dysplastic thymic lobes of drastically reduced size that are ectopically located in the neck at the level of the hyoid bone. Interestingly, the small number of thymocytes in these thymic lobes develops with normal kinetics and shows a wild type phenotype. Organ initiation of the embryonic thymic anlage in these Noggin transgenic mice occurs as in wild-type mice, but the tight temporal and spatial regulation of BMP4 expression is abrogated in subsequent differentiation stages. We show that transgenic Noggin blocks BMP signaling in epithelial as well as mesenchymal cells of the thymic anlage. Our data demonstrate that BMP signaling is crucial for thymus development and that it is the thymic stroma rather than developing thymocytes that depends on BMP signals. PMID- 16210627 TI - KIR3DL1 polymorphisms that affect NK cell inhibition by HLA-Bw4 ligand. AB - The killer cell Ig-like receptor (KIR) gene family encodes MHC class I receptors expressed by NK cells and several T cell subpopulations. Factors contributing to human KIR haplotype diversity are differences in gene number, gene content, and allelic polymorphism. Whereas functional and clinical consequences of the first two factors are established, knowledge of the effects of KIR gene polymorphism is limited to special cases in which signaling function is reversed or cell surface expression lost. In this study we use retrovirally transduced human cell lines to show that 3DL1*002 is a stronger inhibitory receptor for HLA-Bw4 ligands than 3DL1*007. Analysis of mutant 3DL1*002 and 3DL1*007 molecules demonstrates that residue 238 in the D2 domain and 320 in the transmembrane region contribute to the difference in receptor strength. Neither position 238 nor 320 is predicted to interact directly with HLA-Bw4 ligand. This study also revealed that KIR3DL1 and LILRB1 both contribute to developing an inhibitory response to HLA-Bw4 ligands. PMID- 16210628 TI - Unusually high frequency MHC class I alleles in Mauritian origin cynomolgus macaques. AB - Acute shortages of Indian origin Rhesus macaques significantly hinder HIV/AIDS research. Cellular immune responses are particularly difficult to study because only a subset of animals possess MHC class I (MHC I) alleles with defined peptide binding specificities. To expand the pool of nonhuman primates suitable for studies of cellular immunity, we defined 66 MHC I alleles in Cynomolgus macaques (Macaca fascicularis) of Chinese, Vietnamese, and Mauritian origin. Most MHC I alleles were found only in animals from a single geographic origin, suggesting that Cynomolgus macaques from different origins are not interchangeable in studies of cellular immunity. Animals from Mauritius may be particularly valuable because >50% of these Cynomolgus macaques share the MHC class I allele combination Mafa-B*430101, Mafa-B*440101, and Mafa-B*460101. The increased MHC I allele sharing of Mauritian origin Cynomolgus macaques may dramatically reduce the overall number of animals needed to study cellular immune responses in nonhuman primates while simultaneously reducing the confounding effects of genetic heterogeneity in HIV/AIDS research. PMID- 16210629 TI - Diabetes resistance/susceptibility in T cells of nonobese diabetic mice conferred by MHC and MHC-linked genes. AB - Polymorphism of MHC and MHC-linked genes is tightly associated with susceptibility to type 1 diabetes (T1D) in human and animal models. Despite the extensive studies, however, the role of MHC and MHC-linked genes expressed by T cells on T1D susceptibility remains unclear. Because T cells develop from TCR(-) thymic precursor (pre-T) cells that undergo MHC restriction mediated by thymic stroma cells, we reconstituted the T cell compartment of NOD.scid-RIP-B7.1 mice using pre-T cells isolated from NOD, NOR, AKR, and C57BL/6 (B6) mice. T1D developed rapidly in the mice reconstituted with pre-T cells derived from NOD or NOR donors. In contrast, most of the NOD.scid-RIP-B7.1 mice reconstituted with pre-T cells from AKR or B6 donors were free of T1D. Further analysis revealed that genes within MHC locus of AKR or B6 origin reduced incidence of T1D in the reconstituted NOD.scid-RIP-B7.1 mice. The expression of MHC class I genes of k, but not b haplotype, in T cells conferred T1D resistance. Replacement of an interval near the distal end of the D region in T cells of B6 origin with an identical allele of 129.S6 origin resulted in T1D development in the reconstituted mice. These results provide evidence that the expression of MHC class I and MHC-linked genes in T cells of NOD mice indeed contributes to T1D susceptibility, while expression of specific resistance alleles of MHC or MHC linked genes in T cells alone would effectively reduce or even prevent T1D. PMID- 16210630 TI - The murine family of gut-restricted class Ib MHC includes alternatively spliced isoforms of the proposed HLA-G homolog, "blastocyst MHC". AB - The gastrointestinal tract is populated by a multitude of specialized immune cells endowed with receptors for classical (class Ia) and nonclassical (class Ib) MHC proteins. To identify class I products that engage these receptors and impact immunity/tolerance, we studied gut-transcribed class Ib loci and their polymorphism in inbred, outbred, and wild-derived mice. Intestinal tissues enriched in epithelial cells contained abundant transcripts of ubiquitously expressed and preferentially gut-restricted Q and T class I loci. The latter category included the "blastocyst Mhc" gene, H2-Bl, and its putative paralog, Tw5. Expression of H2-Bl was previously detected only at the maternal/fetal interface, where it was proposed to induce immune tolerance via interactions with CD94/NKG2A receptors. Analysis of coding region polymorphism performed here revealed two major alleles of H2-Bl with conserved residues at positions critical for class I protein folding and peptide binding. Both divergent alleles are maintained in outbred and wild mice under selection for fecundity and pathogen resistance. Surprisingly, we found that alternative splicing of H2-Bl mRNA in gut tissues is prevalent and allele-specific. It leads to strain-dependent expression of diverse repertoires of canonical and noncanonical transcripts that may give rise to distinct ligands for intestinal NK cell, T cell, and/or intraepithelial lymphocyte receptors. PMID- 16210631 TI - The RNA helicase Lgp2 inhibits TLR-independent sensing of viral replication by retinoic acid-inducible gene-I. AB - The paramyxovirus Sendai (SV), is a well-established inducer of IFN-alphabeta gene expression. In this study we show that SV induces IFN-alphabeta gene expression normally in cells from mice with targeted deletions of the Toll-IL-1 resistance domain containing adapters MyD88, Mal, Toll/IL-1R domain-containing adaptor inducing IFN-beta (TRIF), and TRIF-related adaptor molecule TLR3, or the E3 ubiquitin ligase, TNFR-associated factor 6. This TLR-independent induction of IFN-alphabeta after SV infection is replication dependent and mediated by the RNA helicase, retinoic acid-inducible gene-I (RIG-I) and not the related family member, melanoma differentiation-associated gene 5. Furthermore, we characterize a RIG-I-like RNA helicase, Lgp2. In contrast to RIG-I or melanoma differentiation associated gene 5, Lgp2 lacks signaling caspase recruitment and activation domains. Overexpression of Lgp2 inhibits SV and Newcastle disease virus signaling to IFN-stimulated regulatory element- and NF-kappaB-dependent pathways. Importantly, Lgp2 does not prevent TLR3 signaling. Like RIG-I, Lgp2 binds double stranded, but not single-stranded, RNA. Quantitative PCR analysis demonstrates that Lgp2 is present in unstimulated cells at a lower level than RIG-I, although both helicases are induced to similar levels after virus infection. We propose that Lgp2 acts as a negative feedback regulator of antiviral signaling by sequestering dsRNA from RIG-I. PMID- 16210632 TI - HoxA10 represses transcription of the gene encoding p67phox in phagocytic cells. AB - p67(phox) and gp91(phox) are components of the phagocyte-specific respiratory burst oxidase that are encoded by the NCF2 and CYBB genes, respectively. These genes are transcribed exclusively in myeloid cells that have differentiated beyond the promyelocyte stage. In mature phagocytes, NCF2 and CYBB transcription continues until cell death and further increases in response to IFN-gamma and other inflammatory mediators. Because p67(phox) and gp91(phox) expression profiles are similar, we hypothesize that common transcription factors interact with homologous cis elements in the CYBB and NCF2 genes to coordinate transcription. Previously, we identified a negative CYBB promoter cis element that is repressed by the homeodomain transcription factor HoxA10. We found that transcriptional repression requires HoxA10-dependent recruitment of histone deacetylase activity to the CYBB cis element. In response to IFN-gamma, phosphorylation of two tyrosine residues in the HoxA10 homeodomain decreases binding to CYBB promoter, thereby abrogating HoxA10-mediated repression. In the current studies, we investigate the possibility that HoxA10 similarly represses NCF2 transcription. We identify a sequence in the NCF2 promoter that is homologous to the HoxA10-binding CYBB cis element. We find that this NCF2 promoter sequence functions as a negative cis element that is repressed by HoxA10 in a tyrosine phosphorylation and histone deacetylase-dependent manner. Our results suggest that cytokine-stimulated pathways regulate HoxA10-mediated repression of the CYBB and NCF2 genes in differentiating myeloid cells and in mature phagocytes during the inflammatory response. Because p67(phox) and gp91(phox) are rate-limiting components for respiratory burst activity, our studies may identify rational therapeutic targets to modulate free radical generation in pathological conditions. PMID- 16210633 TI - Increase in phagocytosis after geldanamycin treatment or heat shock: role of heat shock proteins. AB - The response to injury is activated at the systemic and cellular levels. At the systemic level, phagocytosis plays a key role in controlling infections and clearing necrotic and apoptotic cells. The expression of heat shock proteins (Hsp), which is a well-conserved process, is a major component of cellular response to stress. This study investigated the relationship between Hsps and phagocytosis. An increase in the phagocytosis of opsonized bacteria particles and latex beads was observed upon incubation of murine macrophages with geldanamycin (GA), a specific inhibitor of the Hsp90 family of proteins. The effect of GA on phagocytosis was blocked by coincubation with inhibitors of transcription (actinomycin D) or translation (cycloheximide), suggesting that gene expression was required. Because expression of Hsps has been observed after GA treatment, the effect of heat shock on phagocytosis was investigated. Similar to GA treatment, heat shock resulted in an actinomycin D-sensitive elevation of phagocytosis, which suggests that Hsps are involved. The increase in phagocytosis after GA treatment was not due to increased binding of opsonized particles to their respective receptors on the macrophage surface or to elevated oxidative stress. However, it was correlated with a rapid polymerization of actin in proximity to the plasma membrane. These results suggest that Hsps play a role in the modulation of the phagocytic process, which is part of the stress response. PMID- 16210634 TI - IL-12 is required for induction but not maintenance of protective, memory responses to Blastomyces dermatitidis: implications for vaccine development in immune-deficient hosts. AB - Cellular immunity mediated by T lymphocytes, in particular CD4(+) and CD8(+) type 1 (T1) cells, is the main defense against pathogenic fungi. IL-12 initiates T1 cell development and cell-mediated immunity, but it is unclear whether IL-12 contributes to the maintenance of an antifungal T1 response. In this study, we addressed the role of IL-12 for vaccine-induced memory T cell development against experimental pulmonary blastomycosis. CD4(+) T cells absolutely required IL-12 to control a live genetically engineered attenuated strain of Blastomyces dermatitidis given s.c. as a vaccine, whereas CD8(+) T cells were significantly less dependent on IL-12. Despite differential dependency of T cell subsets on IL 12 during vaccination, neither subset acquired memory immunity in the absence of IL-12. In contrast, adoptive transfer of immune CD4 T cells from wild-type mice into IL-12(-/-) mice showed that CD4(+) T1 memory cells sustained a T1 cytokine profile and remained protective over a period of 6 mo posttransfer. Similarly, memory CD8 cells elicited in IL-12(-/-) mice with killed yeast and transient rIL 12 treatment (during vaccination) remained durable and protective after animals were rested for 3 mo. In conclusion, these studies demonstrate that once CD4 and CD8 cells have acquired a protective T1 phenotype they no longer require the presence of IL-12 to maintain antifungal protective memory. PMID- 16210635 TI - Cloning of the gene encoding a protective Mycobacterium tuberculosis secreted protein detected in vivo during the initial phases of the infectious process. AB - The existence of therapeutic agents and the bacille Calmette-Guerin (BCG) vaccine have not significantly affected the current tuberculosis pandemic. BCG vaccine protects against serious pediatric forms of tuberculosis but not against adult pulmonary tuberculosis, the most common and contagious form of the disease. Several vaccine candidates, including Mycobacterium tuberculosis recombinant proteins formulated in newer adjuvants or delivered in bacterial plasmid DNA have recently been described. An attractive source of vaccine candidates has been M. tuberculosis Ags present in culture supernatants of the initial phases of the bacterial growth in vitro. In this study we describe an Ag discovery approach to select for such Ags produced in vivo during the initial phases of the infection. We combined RP-HPLC and mass spectrometry to identify secreted or shed M. tuberculosis proteins eliminated in animal urine within 14 days after the infection. A peptide containing sequence homology with a hypothetical M. tuberculosis protein was identified and the recombinant protein produced in Escherichia coli. The protein was recognized by Ab (IgG2a and IgG1) and T cells (Th1) of mice infected with M. tuberculosis and by lymphoid cells from healthy donors who had a positive purified protein derivative skin test but not from tuberculosis patients. Moreover, this Ag induced protection in mice against M. tuberculosis at levels comparable to protection induced by BCG vaccine. These results validate the Ag discovery approach of M. tuberculosis proteins secreted or shed in vivo during the early phases of the infection and open new possibilities for the development of potential vaccine candidates or of markers of active mycobacterial multiplication and therefore active disease. PMID- 16210636 TI - Systemic dissemination and persistence of Th2 and type 2 cells in response to infection with a strictly enteric nematode parasite. AB - Oral infection with the nematode parasite Heligmosomoides polygyrus H. polygyrus is entirely restricted to the small intestine. Although the evoked Th2 response has been extensively studied in secondary lymphoid organs, little is known about the systemic dissemination of Th2 cells or type 2 associated eosinophils and basophils. In this study we use bicistronic 4get IL-4 reporter mice to directly visualize the type 2 response to H. polygyrus infection. We observed that CD4(+)/GFP(+) Th2 cells spread systemically and found that these cells accumulated in nonlymphoid "hot spots" in the liver, the lung airways, and the peritoneal cavity. Interestingly, the total number of Th2 cells in the peritoneal cavity was comparable to those found in the draining mesenteric lymph node or the spleen. Peritoneal Th2 cells were distinguished by an exceptionally low apoptotic potential and high expression of the intestinal homing receptor alpha(4)beta(7) integrin. CD4(+)/GFP(+) Th2 cells from these peripheral sites were fully functional as indicated by rapid IL-4 production upon polyclonal or Ag-specific restimulation. Th2 cells persisted in the intestinal tissue and the peritoneal cavity of drug-cured mice for weeks. The presence of peripheral memory Th2 cells in the intestine might be crucial for immunity to recall infections. These findings have important implications for the design of vaccination strategies because it may be necessary to establish and maintain memory CD4(+) T cells at the potential future site of infection. PMID- 16210637 TI - Increased expression of TLR3 in lymph nodes during simian immunodeficiency virus infection: implications for inflammation and immunodeficiency. AB - As pattern recognition receptors, TLRs signal and induce expression of multiple host defense genes including proinflammatory cytokines and chemokines. To investigate the mechanisms of up-regulation of proinflammatory cytokines and chemokines during SIV infection in rhesus macaques, we measured the relative levels of expression of TLRs 1-10 in lymphoid tissues during different stages of SIV infection. By real-time RT-PCR, TLR3 was determined to be up-regulated in macaque lymph nodes (LN) throughout the course of infection, whereas TLR9 was down-regulated during early stages of infection. CXCL9/Mig, CXCL10/IP-10, IFN gamma, and IFN-alpha mRNAs were also increased during acute SIV infection and AIDS. Treatment of macaque spleen and LN cells with TLR3 and TLR9 ligands led to the induction of these same genes. TLR3 stimulation had disparate effects on viral transcription and viral replication, because poly(I:C), a model TLR3 ligand, stimulated the viral promoter but potently inhibited SIV replication in primary cultures of macaque spleen and LN cells. These findings identify roles for TLR3 inflammation in lymphoid tissues and in the immunopathogenesis of HIV 1/SIV, and suggest that TLR3 ligands could potentially be used to flush out latently infected cells that persist during antiretroviral therapies. PMID- 16210638 TI - Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S. AB - We have previously shown that macrophage infection with Mycobacterium tuberculosis and M. bovis bacillus Calmette-Guerin (BCG) partially inhibits MHC class II surface expression in response to IFN-gamma. The present study examined the nature of class II molecules that do in fact reach the surface of infected cells. Immunostaining with specific Abs that discriminate between mature and immature class II populations showed a predominance of invariant chain (Ii) associated class II molecules at the surface of BCG-infected cells suggesting that mycobacteria specifically block the surface export of peptide-loaded class II molecules. This phenotype was due to inhibition of IFN-gamma-induced cathepsin S (Cat S) expression in infected cells and the subsequent intracellular accumulation of alphabeta class II dimers associated with the Cat S substrate Ii p10 fragment. In contrast, infection with BCG was shown to induce secretion of IL 10, and addition of blocking anti-IL-10 Abs to cell cultures restored both expression of active Cat S and export of mature class II molecules to the surface of infected cells. Consistent with these findings, expression of mature class II molecules was also restored in cells infected with BCG and transfected with active recombinant Cat S. Thus, M. bovis BCG exploits IL-10 induction to inhibit Cat S-dependent processing of Ii in human macrophages. This effect results in inhibition of peptide loading of class II molecules and in reduced presentation of mycobacterial peptides to CD4(+) T cells. This ability may represent an effective mycobacterial strategy for eluding immune surveillance and persisting in the host. PMID- 16210639 TI - Mycobacterium tuberculosis up-regulates matrix metalloproteinase-1 secretion from human airway epithelial cells via a p38 MAPK switch. AB - Pulmonary cavitation is vital to the persistence and spread of Mycobacterium tuberculosis (MTb), but mechanisms underlying this lung destruction are poorly understood. Fibrillar type I collagen provides the lung's tensile strength, and only matrix metalloproteinases (MMPs) can degrade it at neutral pH. We investigated MTb-infected lung tissue and found that airway epithelial cells adjacent to tuberculosis (Tb) granulomas expressed a high level of MMP-1 (interstitial collagenase). Conditioned media from MTb-infected monocytes (CoMTb) up-regulated epithelial cell MMP-1 promoter activity, gene expression, and secretion, whereas direct MTb infection did not. CoMTb concurrently suppressed tissue inhibitor of metalloprotease-1 (TIMP-1) secretion, further promoting matrix degradation, and in Tb patients very low TIMP-1 expression was detected. MMP-1 up-regulation required synergy between TNF-alpha and G protein-coupled receptor signaling pathways. CoMTb stimulated p38 MAPK phosphorylation, and this is the point of TNF-alpha synergy with G protein-coupled receptor activation. Furthermore, p38 phosphorylation was the switch up-regulating MMP-1 activity and decreasing TIMP-1 secretion. Activated p38 localized to MMP-1-secreting airway epithelial cells in Tb patients. These data reveal a monocyte-epithelial cell network whereby MTb may drive tissue destruction, and they demonstrate that p38 phosphorylation is a key regulatory point in the generation of a matrix-degrading phenotype. PMID- 16210640 TI - The eotaxin chemokines and CCR3 are fundamental regulators of allergen-induced pulmonary eosinophilia. AB - The eotaxin chemokines have been implicated in allergen-induced eosinophil responses in the lung. However, the individual and combined contribution of each of the individual eotaxins is not well defined. We aimed to examine the consequences of genetically ablating eotaxin-1 or eotaxin-2 alone, eotaxin-1 and eotaxin-2 together, and CCR3. Mice carrying targeted deletions of these individual or combined genes were subjected to an OVA-induced experimental asthma model. Analysis of airway (luminal) eosinophilia revealed a dominant role for eotaxin-2 and a synergistic reduction in eotaxin-1/2 double-deficient (DKO) and CCR3-deficient mice. Examination of pulmonary tissue eosinophilia revealed a modest role for individually ablated eotaxin-1 or eotaxin-2. However, eotaxin-1/2 DKO mice had a marked decrease in tissue eosinophilia approaching the low levels seen in CCR3-deficient mice. Notably, the organized accumulation of eosinophils in the peribronchial and perivascular regions of allergen-challenged wild-type mice was lost in eotaxin-1/2 DKO and CCR3-deficient mice. Mechanistic analysis revealed distinct expression of eotaxin-2 in bronchoalveolar lavage fluid cells consistent with macrophages. Taken together, these results provide definitive evidence for a fundamental role of the eotaxin/CCR3 pathway in eosinophil recruitment in experimental asthma. These results imply that successful blockade of Ag-induced pulmonary eosinophilia will require antagonism of multiple CCR3 ligands. PMID- 16210641 TI - The role of CXCR2/CXCR2 ligand biological axis in renal cell carcinoma. AB - Renal cell carcinoma (RCC) accounts for 3% of new cancer incidence and mortality in the United States. Studies in RCC have predominantly focused on VEGF in promoting tumor-associated angiogenesis. However, other angiogenic factors may contribute to the overall angiogenic milieu of RCC. We hypothesized that the CXCR2/CXCR2 ligand biological axis represents a mechanism by which RCC cells promote angiogenesis and facilitate tumor growth and metastasis. Therefore, we first examined tumor biopsies and plasma of patients with metastatic RCC for levels of CXCR2 ligands, and RCC tumor biopsies for the expression of CXCR2. The proangiogenic CXCR2 ligands CXCL1, CXCL3, CXCL5, and CXCL8, as well as VEGF were elevated in the plasma of these patients and found to be expressed within the tumors. CXCR2 was found to be expressed on endothelial cells within the tumors. To assess the role of ELR(+) CXC chemokines in RCC, we next used a model of syngeneic RCC (i.e., RENCA) in BALB/c mice. CXCR2 ligand and VEGF expression temporally increased in direct correlation with RENCA growth in CXCR2(+/+) mice. However, there was a marked reduction of RENCA tumor growth in CXCR2(-/-) mice, which correlated with decreased angiogenesis and increased tumor necrosis. Furthermore, in the absence of CXCR2, orthotopic RENCA tumors demonstrated a reduced potential to metastasize to the lungs of CXCR2(-/-) mice. These data support the notion that CXCR2/CXCR2 ligand biology is an important component of RCC tumor-associated angiogenesis and tumorigenesis. PMID- 16210642 TI - Characterization of a novel chemokine-containing storage granule in endothelial cells: evidence for preferential exocytosis mediated by protein kinase A and diacylglycerol. AB - We have recently shown that several proinflammatory chemokines can be stored in secretory granules of endothelial cells (ECs). Subsequent regulated exocytosis of such chemokines may then enable rapid recruitment of leukocytes to inflammatory sites. Although IL-8/CXCL8 and eotaxin-3/CCL26 are sorted to the rod-shaped Weibel-Palade body (WPB), we found that GROalpha/CXCL1 and MCP-1/CCL2 reside in small granules that, similarly to the WPB, respond to secretagogue stimuli. In the present study, we report that GROalpha and MCP-1 colocalized in 50- to 100-nm granules, which occur throughout the cytoplasm and at the cell cortex. Immunofluorescence confocal microscopy revealed no colocalization with multimerin or tissue plasminogen activator, i.e., proteins that are released from small granules of ECs by regulated exocytosis. Moreover, the GROalpha/MCP-1-containing granules were Rab27-negative, contrasting the Rab27-positive, WPB. The secretagogues PMA, histamine, and forskolin triggered distinct dose and time dependent responses of GROalpha release. Furthermore, GROalpha release was more sensitive than IL-8 release to inhibitors and activators of PKA and PKC but not to an activator of Epac, a cAMP-regulated GTPase exchange factor, indicating that GROalpha release is regulated by molecular adaptors different from those regulating exocytosis of the WPB. On the basis of these findings, we designated the GROalpha/MCP-1-containing compartment the type 2 granule of regulated secretion in ECs, considering the WPB the type 1 compartment. In conclusion, we propose that the GROalpha/MCP-1-containing type 2 granule shows preferential responsiveness to important mediators of EC activation, pointing to the existence of selective agonists that would allow differential release of selected chemokines. PMID- 16210643 TI - Discovery and pharmacological characterization of a novel rodent-active CCR2 antagonist, INCB3344. AB - This report describes the characterization of INCB3344, a novel, potent and selective small molecule antagonist of the mouse CCR2 receptor. The lack of rodent cross-reactivity inherent in the small molecule CCR2 antagonists discovered to date has precluded pharmacological studies of antagonists of this receptor and its therapeutic relevance. In vitro, INCB3344 inhibits the binding of CCL2 to mouse monocytes with nanomolar potency (IC(50) = 10 nM) and displays dose-dependent inhibition of CCL2-mediated functional responses such as ERK phosphorylation and chemotaxis with similar potency. Against a panel of G protein coupled receptors that includes other CC chemokine receptors, INCB3344 is at least 100-fold selective for CCR2. INCB3344 possesses good oral bioavailability and systemic exposure in rodents that allows in vivo pharmacological studies. INCB3344 treatment results in a dose-dependent inhibition of macrophage influx in a mouse model of delayed-type hypersensitivity. The histopathological analysis of tissues from the delayed-type hypersensitivity model demonstrates that inhibition of CCR2 leads to a substantial reduction in tissue inflammation, suggesting that macrophages play an orchestrating role in immune-based inflammatory reactions. These results led to the investigation of INCB3344 in inflammatory disease models. We demonstrate that therapeutic dosing of INCB3344 significantly reduces disease in mice subjected to experimental autoimmune encephalomyelitis, a model of multiple sclerosis, as well as a rat model of inflammatory arthritis. In summary, we present the first report on the pharmacological characterization of a selective, potent and rodent-active small molecule CCR2 antagonist. These data support targeting this receptor for the treatment of chronic inflammatory diseases. PMID- 16210644 TI - A critical role for complement C3d and the B cell coreceptor (CD19/CD21) complex in the initiation of inflammatory arthritis. AB - Complement C3 cleavage products mediate the recognition and clearance of toxic or infectious agents. In addition, binding of the C3d fragment to Ag promotes B lymphocyte activation through coengagment of the BCR and complement receptor 2 (CD21). Signal augmentation is thought to be achieved through enhanced recruitment and activation of CD21-associated CD19. In this study we show, using the DBA/1 collagen-induced arthritis (CIA) model, that conjugation of C3d to heterologous type II collagen is sufficient to cause disease in the absence of the mycobacterial components of CFA. Transient depletion of C3 during the inductive phase of CIA delays and lessens the severity of disease, and DBA/1 mice deficient for coreceptor components CD19 or CD21 are not susceptible to CIA. Adoptive transfer experiments revealed that CD21 expression on either B cells or follicular dendritic cells is sufficient to acquire disease susceptibility. Although CD19(-/-) and CD21(-/-) mice produce primary Ab responses to heterologous and autologous type II collagen, they are impaired in the ability to activate T cells, form germinal centers, and produce secondary autoantibody responses. These findings indicate that binding of C3d to self-Ags can promote autoimmunity through enhanced Ag retention and presentation by follicular dendritic cells and B cells, respectively. PMID- 16210645 TI - TGF-beta and Smad3 signaling link inflammation to chronic fibrogenesis. AB - Transient adenovirus-mediated gene transfer of IL-1beta (AdIL-1beta), a proinflammatory cytokine, induces marked inflammation and severe and progressive fibrosis in rat lungs. This is associated with an increase in TGF-beta1 concentration in bronchoalveolar lavage (BAL) fluid. TGF-beta1 is a key cytokine in the process of fibrogenesis, using intracellular signaling pathways involving Smad2 and Smad3. In this study we investigate whether inflammation induced by IL 1beta is able to independently induce lung fibrosis in mice deficient in the Smad3 gene. Seven days after AdIL-1beta administration, similar levels of IL 1beta transgene are seen in BAL in both wild-type (WT) and knockout (KO) mice, and BAL cell profiles demonstrated a similar marked neutrophilic inflammation. Phospho-Smad2 staining was positive in areas of inflammation in both WT and KO mice at day 7. By day 35 after transient IL-1beta expression, WT mice showed marked fibrosis in peribronchial areas, quantified by picrosirius red staining and morphometry. However, there was no evidence of fibrosis or collagen accumulation in IL-1beta-treated KO mice, and peribronchial areas were not different from KO mice treated with the control adenovector. TGF-beta1 and phospho-Smad2 were strongly positive at day 35 in fibrotic areas observed in WT mice, but no such staining was detectable in KO mice. The IL-1beta-induced chronic fibrotic response in mouse lungs is dependent on Smad3. KO and WT animals demonstrated a similar inflammatory response to overexpression of IL-1beta indicating that inflammation must link to the Smad3 pathway, likely through TGF beta, to induce progressive fibrosis. PMID- 16210646 TI - Altered CXCR2 signaling in beta-arrestin-2-deficient mouse models. AB - CXCR2 is a G-protein-coupled receptor (GPCR) that binds the CXC chemokines, CXCL1 3 and CXCL5-8, and induces intracellular signals associated with chemotaxis. Many adaptor proteins are actively involved in the sequestration, internalization, and trafficking of CXCR2 and transduction of agonist-induced intracellular signaling. We have previously shown that adaptor protein beta-arrestin-2 (betaarr2) plays a crucial role in transducing signals mediated through CXCR2. To further investigate the role of betaarr2 on CXCR2-mediated signaling during acute inflammation, zymosan-induced neutrophils were isolated from peritoneal cavities of betaarr2-deficient (betaarr2(-/-)) and their wild-type (betaarr2(+/+)) littermate mice, and neutrophil CXCR2 signaling activities were determined by measurement of Ca(2+) mobilization, receptor internalization, GTPase activity, and superoxide anion production. The results showed that the deletion of betaarr2 resulted in increased Ca(2+) mobilization, superoxide anion production, and GTPase activity in neutrophils, but decreased receptor internalization relative to wild-type mice. Two animal models, the dorsal air pouch model and the excisional wound healing model, were used to further study the in vivo effects of betaarr2 on CXCR2-mediated neutrophil chemotaxis and on cutaneous wound healing. Surprisingly, the recruitment of neutrophils was increased in response to CXCL1 in the air pouch model and in the excisional wound beds of betaarr2(-/-) mice. Wound re-epithelialization was also significantly faster in betaarr2(-/-) mice than in betaarr2(+/+) mice. Taken together, the data indicate that betaarr2 is a negative regulator for CXCR2 in vivo signaling. PMID- 16210647 TI - The chemokines CXCL9, CXCL10, and CXCL11 differentially stimulate G alpha i independent signaling and actin responses in human intestinal myofibroblasts. AB - Intestinal myofibroblasts have been implicated in the pathogenesis of chronic inflammatory conditions such as Crohn's disease via interactions with an elaborate network of cytokines, growth factors, and other inflammatory mediators. CXCR3 is a Galpha(i) protein-coupled receptor that binds the proinflammatory chemokines CXCL9, CXCL10, and CXCL11, which are released from the intestinal epithelium. The three CXCR3 ligands shared the ability to activate biochemical (e.g., PI3K and MAPK activation) and functional events (actin reorganization) in intestinal myofibroblasts. However, CXCL11 is unique in its ability to elevate intracellular calcium. Surprisingly, although CXCR3 mRNA is detectable in these myofibroblasts, there is no detectable surface expression of CXCR3. Furthermore, the biochemical responses and actin reorganization stimulated by the CXCR3 ligands in intestinal myofibroblasts are insensitive to the Galpha(i) inhibitor, pertussis toxin. This suggests either the existence of differential receptor coupling mechanisms in myofibroblasts for CXCR3 that are distinct from those observed in PBLs and/or that these cells express a modified or variant CXCR3 compared with the CXCR3 expressed on PBLs. PMID- 16210648 TI - Carboxylated glycans mediate colitis through activation of NF-kappa B. AB - The role of carbohydrate modifications of glycoproteins in leukocyte trafficking is well established, but less is known concerning how glycans influence pathogenesis of inflammation. We previously identified a carboxylate modification of N-linked glycans that is recognized by S100A8, S100A9, and S100A12. The glycans are expressed on macrophages and dendritic cells of normal colonic lamina propria, and in inflammatory infiltrates in colon tissues from Crohn's disease patients. We assessed the contribution of these glycans to the development of colitis induced by CD4(+)CD45RB(high) T cell transfer to Rag1(-/-) mice. Administration of an anti-carboxylate glycan Ab markedly reduced clinical and histological disease in preventive and early therapeutic protocols. Ab treatment reduced accumulation of CD4(+) T cells in colon. This was accompanied by reduction in inflammatory cells, reduced expression of proinflammatory cytokines and of S100A8, S100A9, and receptor for advanced glycation end products. In vitro, the Ab inhibited expression of LPS-elicited cytokines and induced apoptosis of activated macrophages. It specifically blocked activation of NF kappaB p65 in lamina propria cells of colitic mice and in activated macrophages. These results indicate that carboxylate-glycan-dependent pathways contribute to the early onset of colitis. PMID- 16210649 TI - Oxidative stress augments the production of matrix metalloproteinase-1, cyclooxygenase-2, and prostaglandin E2 through enhancement of NF-kappa B activity in lipopolysaccharide-activated human primary monocytes. AB - The excessive production of reactive oxidative species (ROS) associated with inflammation leads to a condition of oxidative stress. Cyclooxygenase-2 (COX-2), PGE(2), and matrix metalloproteinases (MMPs) are important mediators during the process of inflammation. In this paper we report on studies examining how the ROS hydrogen peroxide (H(2)O(2)) affects the production of MMP-1, COX-2, and PGE(2). Addition of H(2)O(2) to LPS-activated monocytes, but not naive monocytes, caused a significant enhancement of the LPS-induced production of MMP-1, COX-2, and PGE(2). The mechanism by which H(2)O(2) increased these mediators was through enhancement of IkappaBalpha degradation, with subsequent increases in NF-kappaB activation and NF-kappaB p50 translocation to the nucleus. The effects of H(2)O(2) on IkappaBalpha degradation, NF-kappaB activation, and NF-kappaB p50 localization to the nucleus were demonstrated through studies of coimmunoprecipitation of IkappaBalpha with p50, ELISA of NF-kappaB p65 activity, and Western blot analysis of the nuclear fraction extract for p50. The key role for NF-kappaB in this process was demonstrated by the ability of MG-132 or lactacystin (proteasome inhibitors) to block the enhanced production of MMP-1, COX-2, and PGE(2). In contrast, indomethacin, which inhibited PGE(2) production, partially blocked the enhanced MMP-1 production. Moreover, although PGE(2) restored MMP-1 production in indomethacin-treated monocyte cultures; it failed to significantly restore MMP-1 production in proteasome inhibitor-treated cultures. Thus, in the presence of LPS and H(2)O(2), NF-kappaB plays a dominate role in the regulation of MMP-1, COX-2, and PGE(2) expression. PMID- 16210650 TI - IL-4 regulates MEK expression required for lysophosphatidic acid-mediated chemokine generation by human mast cells. AB - IL-4 and mast cells (MCs) mediate mucosal defense against helminths and are central to allergic inflammation. Lysophosphatidic acid (LPA), an abundant, potent lipid growth factor, stimulates the growth of cultured human MCs (hMCs) in vitro through a pathway involving LPA receptors 1 and 3 (termed the LPA(1) and LPA(3) receptors, respectively) and peroxisome proliferator-activated receptor gamma. We now report that LPA potently induces the generation of proinflammatory chemokines (MIP-1beta, IL-8, and MCP-1) by hMCs by a mechanism that absolutely requires IL-4. The de novo expression of chemokine mRNA and protein generation involves synergistic actions of calcium flux-dependent NFAT transcription factors and ERK. ERK phosphorylation and chemokine production in response to LPA require IL-4-dependent up-regulation of MEK-1 expression by a pathway involving PI3K. Although receptor-selective agonists for both the LPA(2) and LPA(3) receptors induce calcium fluxes by hMCs, only the LPA(2) receptor-selective agonist fatty alcohol phosphate-12 mimics the IL-4-dependent effect of LPA on chemokine generation. The fact that LPA, an endogenous lipid mediator, activates hMCs by an LPA(2) receptor-dependent pathway indicates functional distinctions between different LPA receptor family members that are expressed constitutively by cells of a single hemopoietic lineage. Moreover, the regulation of MEK-dependent signaling is a mechanism by which IL-4 could amplify inflammation in mucosal immune responses through receptor systems for endogenous ligands such as LPA. PMID- 16210651 TI - Nonpathogenic, environmental fungi induce activation and degranulation of human eosinophils. AB - Eosinophils and their products are probably important in the pathophysiology of allergic diseases, such as bronchial asthma, and in host immunity to certain organisms. An association between environmental fungal exposure and asthma has been long recognized clinically. Although products of microorganisms (e.g., lipopolysaccharides) directly activate certain inflammatory cells (e.g., macrophages), the mechanism(s) that triggers eosinophil degranulation is unknown. In this study we investigated whether human eosinophils have an innate immune response to certain fungal organisms. We incubated human eosinophils with extracts from seven environmental airborne fungi (Alternaria alternata, Aspergillus versicolor, Bipolaris sorokiniana, Candida albicans, Cladosporium herbarum, Curvularia spicifera, and Penicillium notatum). Alternaria and Penicillium induced calcium-dependent exocytosis (e.g., eosinophil-derived neurotoxin release) in eosinophils from normal individuals. Alternaria also strongly induced other activation events in eosinophils, including increases in intracellular calcium concentration, cell surface expression of CD63 and CD11b, and production of IL-8. Other fungi did not induce eosinophil degranulation, and Alternaria did not induce neutrophil activation, suggesting specificity for fungal species and cell type. The Alternaria-induced eosinophil degranulation was pertussis toxin sensitive and desensitized by preincubating cells with G protein coupled receptor agonists, platelet-activating factor, or FMLP. The eosinophil stimulating activity in Alternaria extract was highly heat labile and had an M(r) of approximately 60 kDa. Thus, eosinophils, but not neutrophils, possess G protein-dependent cellular activation machinery that directly responds to an Alternaria protein product(s). This innate response by eosinophils to certain environmental fungi may be important in host defense and in the exacerbation of inflammation in asthma and allergic diseases. PMID- 16210652 TI - T cell mimicry and epitope specificity of cross-reactive T cell clones from rheumatic heart disease. AB - Mimicry between streptococcal M protein and cardiac myosin is important in the pathogenesis of rheumatic heart disease. M protein-specific human T cell clones derived from rheumatic carditis were cross-reactive with human cardiac myosin, and laminin, a valve protein. Among the 11 CD4(+) and CD8(+) cross-reactive T cell clones, at least 6 different reactivity patterns were distinguished, suggesting different degrees of cross-reactivity and a very diverse T cell repertoire. The latter was confirmed by a heterogeneous Vbeta gene and CDR3 usage. HLA restriction and Th1 cytokine production in response to rM6 protein were preserved when the T cell clones were stimulated by human cardiac myosin or other alpha-helical proteins, such as tropomyosin and laminin. The cross-reactive human T cell clones proliferated to B2 and B3A, dominant peptide epitopes in the B repeat region of streptococcal M protein. In human cardiac myosin, epitopes were demonstrated in the S2 and light meromyosin regions. In our study, T cell mimicry was defined as recognition of structurally related Ags involved in disease and recognized by the same T cell. Mimicry in our study was related to alpha-helical coiled coil proteins which have a repetitive seven-aa residue periodicity that maintains alpha-helical structure and thus creates a high number of degenerate possibilities for recognition by T cells. The study of human T cell clones from rheumatic heart disease revealed potential sites of T cell mimicry between streptococcal M protein and human cardiac myosin and represents some of the most well-defined T cell mimicry in human autoimmune disease. PMID- 16210653 TI - A genetic basis for IFN-gamma production and T-bet expression in humans. AB - Th1 and Th2 cytokines secreted by polarized effector T cells play a pivotal role in the development of autoimmune and allergic diseases. However, the genetic basis of cytokine production by T lymphocytes in humans is poorly understood. In this study, we investigated the genetic contribution to cytokine production and regulation of T cell-specific transcription factors in a prospective twin study. We found a substantial genetic contribution to the production of Th1 cytokines such as IFN-gamma and TNF-alpha with heritabilities of 0.85 (95% confidence intervals, 0.74-0.95) and 0.72 (0.50-0.93), respectively, whereas no genetic influence on production of the Th2 signature cytokine IL-4 was observed. Furthermore, the intrapair variability in IFN-gamma production by isolated T cells was lower in monozygotic than in dizygotic twins. In contrast to GATA-3, NFAT, and NF-kappaB, intrapair variability of T-bet, the master transcription factor of Th1 cells, was very low among monozygotic and high among dizygotic twins, indicative of a strong genetic influence on T-bet (heritability 0.93, 95% confidence interval, 0.84-1.0). Our data provide novel insights into the genetic regulation of human Th cell polarization. These data suggest that signature cytokines and cytokine signaling events of Th1 rather than Th2 cells are genetically determined and implicate that Th2-associated diseases in humans might be due to genetic variations in Th1 cytokine regulation via T-bet. This concept is highlighted by the recent finding that inactivation of the T-bet gene in mice results in development of clinical hallmark features of asthma. PMID- 16210654 TI - Human NK cytotoxicity against porcine cells is triggered by NKp44 and NKG2D. AB - Pig-to-human xenotransplantation has been proposed as a means to alleviate the shortage of human organs for transplantation, but cellular rejection remains a hurdle for successful xenograft survival. NK cells have been implicated in xenograft rejection and are tightly regulated by activating and inhibitory receptors recognizing ligands on potential target cells. The aim of the present study was to analyze the role of activating NK receptors including NKp30, NKp44, NKp46, and NKG2D in human xenogeneic NK cytotoxicity against porcine endothelial cells (pEC). (51)Cr release and Ab blocking assays were performed using freshly isolated, IL-2-activated polyclonal NK cell populations as well as a panel of NK clones. Freshly isolated NK cells are NKp44 negative and lysed pEC exclusively in an NKG2D-dependent fashion. In contrast, the lysis of pEC mediated by activated human NK cells depended on both NKp44 and NKG2D, since a complete protection of pEC was achieved only by simultaneous blocking of these activating NK receptors. Using a panel of NK clones, a highly significant correlation between anti-pig NK cytotoxicity and NKp44 expression levels was revealed. Other triggering receptors such as NKp30 and NKp46 were not involved in xenogeneic NK cytotoxicity. Finally, Ab-dependent cell-mediated cytotoxicity of pEC mediated by human NK cells in the presence of xenoreactive Ab was not affected by blocking of activating NK receptors. In conclusion, strategies aimed to inhibit interactions between NKp44 and NKG2D on human NK cells and so far unknown ligands on pEC may prevent direct NK responses against xenografts but not xenogeneic Ab-dependent cell-mediated cytotoxicity. PMID- 16210655 TI - In vivo immunomanipulation of V gamma 9V delta 2 T cells with a synthetic phosphoantigen in a preclinical nonhuman primate model. AB - Vgamma9Vdelta2(+) cells represent the major population of gammadelta T cells in primate blood and react in an MHC-unrestricted fashion to a set of low m.w. nonpeptide phosphoantigens. Two types of structurally related agonists have been discovered so far: the natural phosphoantigens (hydroxydimethyl allyl pyrophosphate or isopentenyl-pyrophosphate (IPP)) acting directly on Vgamma9Vdelta2(+) TCR and aminobisphosphonates, which block the mevalonate pathway in target cells, leading to accumulation of natural phosphoantigens that in turn activate Vgamma9Vdelta2(+) cells. We demonstrate in the cynomolgus monkey that Vgamma9Vdelta2 can be manipulated in vivo with bromohydrin pyrophosphate (BrHPP)/Phosphostim, a potent synthetic agonist for which the mechanism of action is similar to natural phosphoantigens. Although of very short half-life, injection of BrHPP leads to strong activation of Vgamma9Vdelta2, inducing production of a high level of Th1 cytokines. Combination of BrHPP with low-dose rhIL-2 induces specific amplification of effector-memory peripheral Vgamma9Vdelta2 in blood in a dose-dependant manner. This transient response returns to baseline within 10-15 days. Successive infusions of BrHPP and rhIL-2 induce less vigorous expansions, suggesting a progressive exhaustion of the response. As no toxicity is detected with or without IL-2, this scheme represents a promising immunotherapeutic strategy for induction of systemic Th1 cytokines and massive expansion of gammadelta T cell subset with antitumor and anti infectious properties. PMID- 16210656 TI - V gamma 9V delta 2 T cell response to colon carcinoma cells. AB - During analysis of CD8 T cells derived from ascites of a colon cancer patient, we isolated a Vgamma9Vdelta2 T cell clone showing strong reactivity against autologous tumor cell lines. This clone killed a large fraction of allogeneic colon carcinoma and melanoma cell lines, but did not affect a normal colon cell line, colon fibroblasts, or melanocytes. Tumor cell recognition was TCR and NKG2D dependent and induced TNF-alpha and IFN-gamma secretion by the clone; accordingly, tumor targets expressed several NKG2D ligands, such as MHC class I chain-related gene A and UL16-binding protein molecules. Colon tumor recognition by Vgamma9Vdelta2 T cells was highly dependent on isopentenyl pyrophosphate production and ICAM-1 expression by target cells. Finally, similar reactivity patterns against colon carcinoma cell lines were observed using polyclonal Vgamma9Vdelta2 T cells of various origins, and Vgamma9Vdelta2 lymphocytes were present in the majority of colon tumor samples studied. Together, these results suggest that Vgamma9Vdelta2 T cells contribute to the natural immune surveillance against colon cancers. Therefore, this study provides a strong rationale for the use of Vgamma9Vdelta2 T cell agonists in immunotherapies targeting colon tumors. PMID- 16210657 TI - The control of allergic conjunctivitis by suppressor of cytokine signaling (SOCS)3 and SOCS5 in a murine model. AB - Allergic conjunctivitis (AC) is a common allergic eye disease characterized by clinical symptoms such as itchiness, conjunctival congestion, elevated Ag specific IgE, mast cell activation, and local eosinophil infiltration. In this study we established a murine model for Ag-induced AC to understand the pathogenesis of the disease. Cell transfer experiments indicated that AC can be divided into early and late phase responses (EPR and LPR). EPR was associated with IgE responses, leading to itchiness, whereas LPR was characterized by local eosinophil infiltration. Both EPR and LPR were significantly inhibited in STAT6 deficient mice, and adoptive transfer of Th2 cells reconstituted LPR. Furthermore, SOCS3 was highly expressed at the disease site, and T cell-specific expression of SOCS3 deteriorated clinical and pathological features of AC, indicating that Th2-mediated SOCS3 expression controls the development and persistence of AC. Reduction of the expression level in SOCS3 heterozygous mice or inhibition of function in dominant-negative SOCS3 transgenic mice clearly reduced the severity of AC. In contrast, constitutive expression of SOCS5, a specific inhibitor of IL-4 signaling, resulted in reduced eosinophil infiltration. These results suggest that negative regulation of the Th2-mediated response by dominant-negative SOCS3 and SOCS5 could be a target for therapeutic intervention in allergic disease. PMID- 16210658 TI - Defined blocks in terminal plasma cell differentiation of common variable immunodeficiency patients. AB - Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by defective Ab production and recurrent bacterial infections. The largely unknown causes are likely to comprise a diverse set of genetic or acquired defects. In this study, we investigated terminal B cell differentiation in lymph nodes from CVID patients. Up to the germinal center B cell stage, B cell differentiation was normal but terminal plasma cell development was found to be impaired. Using differential Blimp-1 and Syndecan-1 expression in controls, we defined three different plasma cell subsets that correspond to progressive developmental stages locating to different sites in the lymph node. In the CVID patients, we could only detect one or two of these subsets indicating a defective differentiation. Thus, terminal plasma cell differentiation was found to be impaired despite normal expression of Blimp-1. B cells reaching only the first stage of plasma cell differentiation were further unable to undergo isotype switching and to up-regulate activation markers on B cells stimulated in vitro. PMID- 16210660 TI - Treatment with a laminin-derived peptide suppresses lupus nephritis. AB - The role of DNA as the target for pathogenic lupus autoantibodies in systemic lupus erythematosus is equivocal and renal damage may be due to cross-reactivity of lupus Abs with glomerular components. We have previously shown that lupus autoantibodies bind to the laminin component of the extracellular matrix. In the present work, we have analyzed the fine specificity of the interaction of pathogenic murine lupus autoantibodies with this molecule and the effect of inhibiting their binding to laminin during the course of the disease. We have found that pathogenic murine lupus autoantibodies react with a 21-mer peptide located in the globular part of the alpha-chain of laminin. Immunization of young lupus-prone mice with this peptide accelerated renal disease. Analysis of transgenic, congenic, and RAG-1(-/-) mice confirmed the importance of this epitope in the pathogenesis of lupus renal disease. We have synthesized a panel of peptides that cross-react with the anti-laminin Abs and have found that the binding of lupus autoantibodies to the extracellular matrix could be inhibited in vitro by some of these competitive peptides. Treatment of MRL/lpr/lpr mice with these peptides prevented Ab deposition in the kidneys, ameliorated renal disease, and prolonged survival of the peptide-treated mice. We suggest that laminin components can serve as the target for lupus Abs. The interaction with these Ags can explain both the tissue distribution and the immunopathological findings in lupus. Moreover, inhibition of autoantibody binding to the extracellular matrix can lead to suppression of disease. PMID- 16210659 TI - HLA-A*0201, HLA-A*1101, and HLA-B*0702 transgenic mice recognize numerous poxvirus determinants from a wide variety of viral gene products. AB - In virus models explored in detail in mice, CTL typically focus on a few immunodominant determinants. In this study we use a multipronged approach to understand the diversity of CTL responses to vaccinia virus, a prototypic poxvirus with a genome approximately 20-fold larger than that of the model RNA viruses typically studied in mice. Based on predictive computational algorithms for peptide binding to HLA supertypes, we synthesized a panel of 2889 peptides to begin to create an immunomic map of human CTL responses to poxviruses. Using this panel in conjunction with CTLs from vaccinia virus-infected HLA transgenic mice, we identified 14 HLA-A*0201-, 4 HLA-A*1101-, and 3 HLA-B*0702-restricted CD8(+) T cell determinants distributed over 20 distinct proteins. These peptides were capable of binding one or multiple A2, A3, and B7 supertype molecules with affinities typical of viral determinants. Surprisingly, many of the viral proteins recognized are predicted to be late gene products, in addition to the early intermediate gene products expected. Nearly all of the determinants identified have identical counterparts encoded by modified vaccinia virus Ankara as well as variola virus, the agent of smallpox. These findings have implications for the design of new smallpox vaccines and the understanding of immune responses to large DNA viruses in general. PMID- 16210661 TI - Requirement for neutralizing antibodies to control bone marrow transplantation associated persistent viral infection and to reduce immunopathology. AB - Bone marrow transplantation (BMT) is commonly used in the treatment of leukemia, however its therapeutic application is partly limited by the high incidence of associated opportunistic infections. We modeled this clinical situation by infecting mice that underwent BMT with lymphocytic choriomeningitis virus (LCMV) and investigated the potential of immunotherapeutic strategies to counter such infections. All mice that received BMT survived LCMV infection and developed a virus carrier status. Immunotherapy by adoptive transfer of naive splenocytes protected against low (200 PFU), but not high (2 x 10(6) PFU), doses of LCMV. Attempts to control infection of high viral titers using strongly elevated frequencies of activated LCMV-specific T cells failed to control virus and resulted in immunopathology and death. In contrast, virus neutralizing Abs combined with naive splenocytes were able to efficiently control high-dose LCMV infection without associated side effects. Thus, cell transfer combined with neutralizing Abs represented the most effective means of controlling BMT associated opportunistic viral infection in our in vivo model. These data underscore the in vivo efficacy and immunopathological "safety" of neutralizing antibodies. PMID- 16210662 TI - IL-13 acutely augments HIV-specific and recall responses from HIV-1-infected subjects in vitro by modulating monocytes. AB - We show in this study that acute exposure of PBMCs derived from HIV-infected subjects to IL-13 results in increased recall T cell lymphoproliferative responses against HIV-1 p24 (n = 30, p < 0.0001) and other recall Ags (influenza, n = 43, p < 0.0001; purified protein derivative tuberculin, n = 6, p = 0.0299). This effect is due to a mechanism that acutely targets APC function in the adherent monocyte subset, as shown by the expansion of CD4(+) T cell responses following coculture of IL-13-treated enriched CD14(+) monocytes with donor matched enriched CD4(+) T cells and Ag. Exposure to IL-13 over 18-72 h resulted in a significant enhancement of monocyte endocytosis (n = 11, p = 0.0005), CD86 expression (n = 12, p = 0.001), and a significant decrease in spontaneous apoptosis (n = 8, p = 0.008). Moreover, IL-13 exposure induced a significant decrease of significantly elevated constitutive levels of PBMC-secreted TNF-alpha (n = 14, p < 0.001) and IL-10 (n = 29, p < 0.001) within 18 h of exposure ex vivo, also reflected by decreased gene expression in the adherent cell population. Our data show that IL-13 is able to acutely enhance the function of the CD14(+) cell subset toward supporting Ag-specific cell-mediated responses in chronic HIV-1 infection. PMID- 16210663 TI - Inhibition of NK cell activity through TGF-beta 1 by down-regulation of NKG2D in a murine model of head and neck cancer. AB - In an orthotopic murine model of head and neck squamous cell carcinoma (SCC VII/SF) we studied NK cell-mediated immunity following vaccination with a recombinant vaccinia virus expressing IL-2 (rvv-IL-2). SCC VII/SF tumor cells were injected into the oral cavity of C3H/HeJ mice on day 0. Mice were vaccinated on days 7, 10, and 14 with rvv-IL-2 and control vaccines. Phenotypes, numbers, and biological activities of NK cells were determined following vaccination. Levels of expression of NK-activating receptor NKG2D and CD16 on NK cell surface were assayed in the vaccinated mice. Expression of NKG2D ligands, Rae1, and H60 on SCC VII/SF cells was also examined. Vaccination with rvv-IL-2 resulted in expansion of NK cells. NK cells isolated from rvv-IL-2-vaccinated mice had significantly higher biological activities compared with mice treated with control vaccines. NK cells from tumor-bearing mice expressed significantly lower levels of NKG2D and CD16 compared with rvv-IL-2 vaccinated mice. SCC VII/SF tumors expressed NKG2D ligand Rae 1, although H60 was not present. SCC VII/SF tumors expressed high levels of TGF-beta1, which were down-modulated by vaccination with rvv-IL-2. Incubation of NK cells with tumor homogenate or cultured supernatant of SCC VII/SF cells reduced the expression of NKG2D and CD16. This inhibition appeared to be mediated by TGF-beta1. SCC VII/SF tumors in the oral cavity of the mice secrete high quantities of TGF-beta1, which reduce the expression of NK cell receptor NKG2D as well as CD16 and inhibits biological functions of NK cells. PMID- 16210665 TI - Omega-3 fatty acid prevents heart rate variability reductions associated with particulate matter. AB - CONTEXT: Environmental exposure to particulate matter of 2.5 microm or less (PM2.5) has been associated with changes in heart rate variability (HRV). OBJECTIVE: To evaluate the effect of supplementation with omega-3 polyunsaturated fatty acids on the reduction of HRV associated with PM2.5 exposure. DESIGN: Randomized double-blind trial. SETTING: Mexico City, Mexico. PARTICIPANTS: 50 nursing home residents older than 60 yr. INTERVENTION: Randomization to either 2 g/d of fish oil versus 2 g/d of soy oil as the control, with 6 mo follow-up (1-mo presupplementation and 5-mo supplementation) or repeated HRV measurements. PM2.5 was monitored indoors and outdoors. MAIN OUTCOME MEASURE: The association between HRV and 1 SD change in PM2.5 (8 microg/m3). RESULTS: In the group receiving fish oil, the reduction in HRV-high-frequency log(10)-transformed associated with a 1 SD change in PM2.5 was -54% (95% confidence interval, -72, -24) in the presupplementation phase, and only -7% (95% confidence interval, -20,+7) in the supplementation phase (p < 0.01 for the effect of supplementation), with changes in other HRV parameters also being significantly less pronounced during supplementation. Small decreases in PM2.5-associated reductions in HRV parameters also occurred in the group receiving soy oil, but these were not significant. Fish oil supplementation was significantly better in preventing the reduction in percentage of successive normal RR intervals differing by more than 50 ms (p = 0.03) and the root square of the mean of the sum of the squares of differences between adjacent intervals (p = 0.05) than soy oil supplementation. INTERPRETATION: Supplementation with 2 g/d of fish oil prevented HRV decline related to PM2.5 exposure in the study population. PMID- 16210664 TI - Mycobacterium tuberculosis growth control by lung macrophages and CD8 cells from patient contacts. AB - RATIONALE: Healthy household contacts (HHCs) of patients with active pulmonary tuberculosis are exposed aerogenically to Mycobacterium tuberculosis (Mtb), thus permitting the study of protective local immunity. OBJECTIVES: To assess alveolar macrophage (AM) and autologous blood CD4 and CD8 T-cell-mediated Mtb growth control in HHCs and healthy, unexposed community control subjects (CCs). METHODS: AMs were infected with Mtb strains H(37)Ra and H(37)Rv at multiplicities of infection 0.1 and 1. Mtb colony-forming units were evaluated on Days 1, 4, and 7. MAIN RESULTS: CD8 T cells from HHCs in 1:1 cocultures with AMs significantly (p < 0.05) increased Mtb growth control by AMs. In CCs, no detectable contribution of CD8 T cells to Mtb growth control was observed. CD4 T cells did not increase Mtb growth control in HHCs or in CCs. IFN-gamma, nitric oxide, and tumor necrosis factor were determined as potential mediators of Mtb growth control in AMs and AM/CD8 and AM/CD4 cocultures. IFN-gamma production in AM/CD4 was twofold higher than that in AM/CD8 cocultures in both HHCs and CCs (p < 0.05). Nitric oxide production from AMs of HHCs increased on Days 4 and 7 and was undetectable in AMs from CCs. IFN-gamma and nitric acid concentrations and Mtb growth control were not correlated. Tumor necrosis factor levels were significantly increased in AM/CD8 cocultures from HHCs compared with AM/CD8 cocultures from CCs (p < 0.05). CONCLUSION: Aerogenic exposure to Mtb in HHCs leads to expansion of Mtb-specific effector CD8 T cells that limit Mtb growth in autologous AMs. PMID- 16210667 TI - Mast cells protect mice from Mycoplasma pneumonia. AB - RATIONALE: As the smallest free-living bacteria and a frequent cause of respiratory infections, mycoplasmas are unique pathogens. Mice infected with Mycoplasma pulmonis can develop localized, life-long airway infection accompanied by persistent inflammation and remodeling. OBJECTIVE: Because mast cells protect mice from acute septic peritonitis and gram-negative pneumonia, we hypothesized that they defend against mycoplasma infection. This study tests this hypothesis using mast cell-deficient mice. METHODS: Responses to airway infection with M. pulmonis were compared in wild-type and mast cell-deficient Kit(W-sh)/Kit(W-sh) mice and sham-infected control mice. MEASUREMENTS AND MAIN RESULTS: Endpoints include mortality, body and lymph node weight, mycoplasma antibody titer, and lung mycoplasma burden and histopathology at intervals after infection. The results reveal that infected Kit(W-sh)/Kit(W-sh) mice, compared with other groups, lose more weight and are more likely to die. Live mycoplasma burden is greater in Kit(W-sh)/Kit(W-sh) than in wild-type mice at early time points. Four days after infection, the difference is 162-fold. Titers of mycoplasma-specific IgM and IgA appear earlier and rise higher in Kit(W-sh)/Kit(W-sh) mice, but antibody responses to heat-killed mycoplasma are not different compared with wild type mice. Infected Kit(W-sh)/Kit(W-sh) mice develop larger bronchial lymph nodes and progressive pneumonia and airway occlusion with neutrophil-rich exudates, accompanied by angiogenesis and lymphangiogenesis. In wild-type mice, pneumonia and exudates are less severe, quicker to resolve, and are not associated with increased angiogenesis. CONCLUSIONS: These findings suggest that mast cells are important for innate immune containment of and recovery from respiratory mycoplasma infection. PMID- 16210668 TI - Family aggregation of upper airway soft tissue structures in normal subjects and patients with sleep apnea. AB - RATIONALE: Sleep apnea is believed to be a genetic disorder. Thus, we hypothesized that anatomic risk factors for sleep apnea would demonstrate family aggregation. OBJECTIVES: We used volumetric magnetic resonance imaging in a sib pair "quad" design to study the family aggregation of the size of upper airway soft tissue structures that are associated with increased risk for obstructive sleep apnea. METHODS: We examined 55 sleep apnea probands (apnea-hypopnea index [AHI]: 43.2 +/- 26.3 events/h), 55 proband siblings (AHI: 11.8 +/- 16.6 events/h), 55 control subjects (AHI: 2.1 +/- 1.7 events/h), and 55 control siblings (AHI: 4.2 +/- 4.0 events/h). The study design used exact matching on ethnicity and sex, frequency matching on age, and statistical control for visceral neck fat and craniofacial dimensions. MEASUREMENTS AND MAIN RESULTS: The data support our a priori hypothesis that the volume of the important upper airway soft tissue structures is heritable. The volume of the lateral pharyngeal walls (h(2) = 36.8%; p = 0.001), tongue (h(2) = 36.5%; p = 0.0001), and total soft tissue (h(2) = 37.5%; p = 0.0001) demonstrated significant levels of heritability after adjusting for sex, ethnicity, age, visceral neck fat, and craniofacial dimensions. In addition, our data indicate that heritability of the upper airway soft tissue structures is found in normal subjects and patients with apnea. Thus, it is not simply a consequence of the prevalence of apnea. CONCLUSIONS: This is the first time family aggregation of size of the upper airway soft tissue structures has been demonstrated. PMID- 16210669 TI - The NHLBI lymphangioleiomyomatosis registry: characteristics of 230 patients at enrollment. AB - RATIONALE: Pulmonary lymphangioleiomyomatosis is a progressive cystic lung disease that is associated with infiltration of atypical smooth muscle-like cells. Previous descriptions of clinical characteristics of subjects with lymphangioleiomyomatosis have been based on a limited number of patients. OBJECTIVES: To describe the clinical characteristics of subjects with pulmonary lymphangioleiomyomatosis, both sporadic and tuberous sclerosis-related forms. METHODS: Over a 3-yr period, from 1998 to 2001, 243 subjects with pulmonary lymphangioleiomyomatosis were enrolled into a national registry; 13 subjects who had already undergone lung transplantation were excluded for the purposes of this report. MEASUREMENTS AND MAIN RESULTS: All 230 subjects were women, aged 18 to 76 yr (mean +/- SE, 44.5 +/- 0.65 yr). The average age at onset of symptoms was 38.9 +/- 0.73 yr and at diagnosis was 41.0 +/- 0.65 yr. Tuberous sclerosis complex was present in 14.8% of subjects. Pulmonary manifestations, most commonly spontaneous pneumothorax, were the primary events leading to the diagnosis in 86.5% of cases. Nearly 55% of the subjects were being treated with a progesterone derivative. An obstructive pattern on pulmonary function testing was observed in 57.3% of the subjects, whereas 33.9% had normal spirometric results. Women with tuberous sclerosis-related lymphangioleiomyomatosis were younger and had less impaired lung function compared with those with the sporadic form. CONCLUSIONS: The age range of women afflicted with pulmonary lymphangioleiomyomatosis is broader than previously appreciated and the degree of pulmonary function can be quite variable, with one-third of subjects having normal spirometry at enrollment into this registry. PMID- 16210670 TI - Growth of pulmonary microvasculature in ventilated preterm infants. AB - RATIONALE: Density-based morphometric studies have demonstrated decreased capillary density in infants with bronchopulmonary dysplasia (BPD) and in BPD like animal models, leading to the prevailing view that microvascular development is disrupted in BPD. OBJECTIVE: To perform a comprehensive analysis of the early and late effects of ventilation on pulmonary microvascular growth in preterm infants. METHODS: Postmortem lung samples were collected from ventilated preterm infants who died between 23 and 29 wk ("short-term ventilated") or between 36 and 39 wk ("long-term ventilated") corrected postmenstrual age. Results were compared with age-matched infants or stillborn infants ("early" and "late" control subjects). Microvascular growth was studied by anti-platelet endothelial cell adhesion molecule (PECAM)-1 immunohistochemistry, quantitative stereology, analysis of endothelial cell proliferation, and Western blot analysis of pulmonary PECAM-1 protein levels. MEASUREMENTS: Measurements were made of capillary density, volume of air-exchanging parenchyma, volume of microvascular endothelial cells, Ki67 labeling index of endothelial cells, and PECAM-1/actin protein levels. MAIN RESULTS: Lungs of long-term ventilated infants showed a significant (more than twofold) increase in volume of air-exchanging parenchyma and a 60% increase in total pulmonary microvascular endothelial volume compared with late control subjects, associated with 60% higher pulmonary PECAM-1 protein levels. The marked expansion of the pulmonary microvasculature in ventilated lungs was, at least partly, attributable to brisk endothelial cell proliferation. The microvasculature of ventilated lungs appeared immature, retaining a saccular architectural pattern. CONCLUSIONS: The pulmonary microvasculature of ventilated preterm infants displayed marked angiogenesis, nearly proportionate to the growth of the air-exchanging lung parenchyma. These results challenge the paradigm of microvascular growth arrest as a major pathogenic factor in BPD. PMID- 16210671 TI - Leptin corrects host defense defects after acute starvation in murine pneumococcal pneumonia. AB - RATIONALE: Leptin is an adipocyte-derived hormone that declines dramatically during fasting and plays a pivotal role in the neuroendocrine response to starvation. Previously, we employed leptin-deficient (ob/ob) mice to identify an important role for leptin in the host defense against Klebsiella pneumonia. OBJECTIVES: To assess the effects of fasting on the innate immune response against pneumococcal pneumonia and to determine the effects of maintaining circulating leptin levels on host defense in fasted mice. METHODS: C57BL/6 mice were either fed ad libitum or fasted for 48 h and given an intraperitoneal injection of saline or recombinant leptin (1 microg/g of body weight) twice daily for 48 h before bacterial challenge. Mice were challenged with 10(5) cfu of Streptococcus pneumoniae via the intranasal route. MEASUREMENTS AND MAIN RESULTS: Lung homogenate S. pneumoniae burden was nearly 20-fold greater in the fasted as compared with fed mice. The impairment in bacterial clearance observed in fasted animals was associated with reduced bronchoalveolar lavage neutrophil counts and interleukin-6 and macrophage inflammatory protein-2 levels. Alveolar macrophages from fasted animals also exhibited defective phagocytosis and killing of S. pneumoniae and reduced calcium-ionophore-stimulated leukotriene B(4) synthesis in vitro. In contrast, the provision of exogenous leptin to fasted animals restored bacterial clearance, bronchoalveolar lavage levels of neutrophils and cytokines, alveolar macrophage bacterial killing, and leukotriene B(4) synthesis. CONCLUSIONS: These results suggest that reduced leptin levels substantially contribute to the suppression of pulmonary antibacterial host defense during starvation and that administration of this adipokine may be of therapeutic benefit clinically. PMID- 16210666 TI - Asthma in Hispanics. AB - Hispanic individuals trace their ancestry to countries that were previously under Spanish rule, including Mexico, large parts of Central and South America, and some Caribbean islands. Most--but not all--Hispanics have variable proportions of European, Amerindian, and African ancestry. Hispanics are diverse with regard to many factors, including racial ancestry, country of origin, area of residence, socioeconomic status, education, and access to health care. Recent findings suggest that there is marked variation in the prevalence, morbidity, and mortality of asthma in Hispanics in the United States and in Hispanic America. The reasons for differences in asthma and asthma morbidity among and within Hispanic subgroups are poorly understood but are likely due to the interaction between yet-unidentified genetic variants and other factors, including environmental tobacco smoke exposure, obesity, allergen exposure, and availability of health care. Barriers to optimal management of asthma in Hispanics in the United States and in Hispanic America include inadequate access to health care, suboptimal use of antiinflammatory medications, and lack of reference values for spirometric measures of lung function in many subgroups (e.g., Puerto Ricans). Future studies of asthma in Hispanics should include large samples of subgroups that are well characterized with regard to self-reported ethnicity, country of origin, place of birth, area of residence, and indicators of socioeconomic status. Because Hispanics are disproportionately represented among the poor in the United States, implementation of adequate access to health care and social reforms (e.g., improving housing conditions) would likely have a major impact on reducing asthma morbidity in this population. PMID- 16210672 TI - Differential roles of CD14 and toll-like receptors 4 and 2 in murine Acinetobacter pneumonia. AB - RATIONALE: Acinetobacter baumannii is an opportunistic bacterial pathogen that is increasingly associated with gram-negative nosocomial pneumonia, but the molecular mechanisms that play a role in innate defenses during A. baumannii infection have not been elucidated. OBJECTIVE: To gain first insight into the role of CD14 and Toll-like receptors 4 and 2 in host response to A. baumannii pneumonia. METHODS: Respective gene-deficient mice were intranasally infected with A. baumannii, and bacterial outgrowth, lung inflammation, and pulmonary cytokine/chemokine responses were determined. To study the importance of LPS in the inflammatory response, mice were also challenged with A. baumannii LPS. MEASUREMENTS AND MAIN RESULTS: Bacterial counts were increased in CD14 and Toll like receptor 4 gene-deficient mice, and only these animals developed bacteremia. The pulmonary cytokine/chemokine response was impaired in Toll-like receptor 4 knockout mice and the onset of lung inflammation was delayed. In contrast, Toll like receptor 2-deficient animals displayed an earlier cell influx into lungs combined with increased macrophage inflammatory protein-2 and monocyte chemoattractant protein-1 concentrations, which was associated with accelerated elimination of bacteria from the pulmonary compartment. Neither CD14 nor Toll like receptor 4 gene-deficient mice responded to intranasal administration of LPS, whereas Toll-like receptor 2 knockout mice were indistinguishable from wild type animals. CONCLUSIONS: Our results suggest that CD14 and Toll-like receptor 4 play a key role in innate sensing of A. baumannii via the LPS moiety, resulting in effective elimination of the bacteria from the lung, whereas Toll-like receptor 2 signaling seems to counteract the robustness of innate responses during acute A. baumannii pneumonia. PMID- 16210673 TI - Genomic profiling of cancer: what next? PMID- 16210674 TI - Persistent residual disease in t(11;18)(q21;q21) positive gastric mucosa associated lymphoid tissue lymphoma treated with chemotherapy or rituximab. PMID- 16210675 TI - Chemosensitivity and chemoresistance testing. PMID- 16210678 TI - Physician recommendation for colorectal cancer screening by race, ethnicity, and health insurance status among men and women in the United States, 2000. AB - In this study, the authors examined whether men and women in the United States had received a physician recommendation to get a colorectal cancer screening test, by race, ethnicity, and health insurance status using data from the 2000 National Health Interview Survey. Among men and women who had had a doctor visit in the past year but who had not had a recent fecal occult blood test, about 94.6% (95% CI 94.0-95.2) reported that their doctor had not recommended the test in the past year. African Americans, Hispanics, and American Indians/Alaska Natives were less likely to report receiving a recommendation for endoscopy compared to Whites. PMID- 16210679 TI - Considerations in recruiting underscreened women to focus groups on screening for cervical cancer. AB - This article describes recruitment activities and costs from two independently conducted studies that used similar, systematic approaches to recruiting two subgroups of underscreened women, Black women and Mexican women. The studies varied in number of recruiters, venues of recruitment, and region of the country. The ratio of women approached to women who were underscreened was 4:1 for Black women and 10:1 for Mexican women. Hysterectomy was a predominant reason for ineligibility among Black women but not Mexican women. In both studies, personal networks were the most productive method of identifying women. Flyers and organized community venues were least productive. The cost incurred for identifying a woman who was eligible for a focus group was 145 dollars for Black women and 59 dollars for Mexican women. Those planning research or program activities that include recruiting underscreened women either to focus groups or health services could benefit from this information. PMID- 16210680 TI - Developing English and Spanish television public service announcements to promote colorectal cancer screening. AB - Screen for Life: National Colorectal Cancer Action Campaign (SFL) is a federally funded campaign to promote understanding of colorectal cancer and the importance of regular screening for adults aged 50 years and older. SFL uses a variety of communication strategies, including television public service announcements (PSAs). SFL materials are developed using the Health Communication Process endorsed by the U.S. Department of Health and Human Services, which has four stages: (a) planning and strategy development; (b) developing and pretesting concepts, messages, and materials; (c) implementing the program; and (d) assessing effectiveness and making refinements. This article describes SFL's application of this process to develop television PSAs in English and Spanish. PMID- 16210681 TI - Using intervention mapping to develop a breast and cervical cancer screening program for Hispanic farmworkers: Cultivando La Salud. AB - This article describes the development of the Cultivando La Salud program, an intervention to increase breast and cervical cancer screening for Hispanic farmworker women. Processes and findings of intervention mapping (IM), a planning process for development of theory and evidence-informed program are discussed. The six IM steps are presented: needs assessment, preparation of planning matrices, election of theoretic methods and practical strategies, program design, implementation planning, and evaluation. The article also describes how qualitative and quantitative findings informed intervention development. IM helped ensure that theory and evidence guided (a) the identification of behavioral and environmental factors related to a target health problem and (b) the selection of the most appropriate methods and strategies to address the identified determinants. IM also guided the development of program materials and implementation by lay health workers. Also reported are findings of the pilot study and effectiveness trial. PMID- 16210682 TI - Development of a linguistically and culturally appropriate booklet for Latino cancer survivors: lessons learned. AB - In response to the need for linguistically and culturally appropriate cancer survivorship materials for Latinos, the Office of Education and Special Initiatives and the Office of Cancer Survivorship at the National Cancer Institute (NCI) set out to test, adapt, and refine a Spanish translation of an English-language booklet for adult cancer survivors titled Facing Forward: Life After Cancer Treatment (Siga adelante: la vida despues del tratamiento del cancer). The authors used a process called "transcreation," which involves translating existing English-language materials into Spanish and then adapting them for Latino audiences. The Spanish version of the booklet was reviewed by nine Spanish-speaking reviewers who were cancer survivors. The multistage transcreation process reinforced the importance of conducting formative research as well as adjusting the methodology to address the needs of changing demographics. PMID- 16210683 TI - Development of an interviewer training manual for a cervical health project on the Apsaalooke Reservation. AB - This article describes strategies used to develop a survey interview training manual for use on the Apsaalooke (Crow Indian) Reservation and delineates how this process and product differed from those discussed in the extant literature on survey interview training. Working to ensure cultural appropriateness is especially important due to past research improprieties with Native American populations. This manual was developed as a part of a cervical health intervention program, Messengers for Health. Areas covered include goals of survey research, recruitment and enrollment, manner of the interviewer, nonverbal behavior, beginning the interview, and language use. Limitations of this work and suggestions for conducting survey research with Native American populations are also included. PMID- 16210684 TI - When bioterrorism strikes: communication issues for the local health department. AB - Public health preparedness is a multifaceted planning process that becomes grounded in a response plan and in effective communications, internal and external. This article describes an incident when the presence of anthrax spores was detected in a postal facility within Kansas City, Missouri, and discusses the communications issues faced by the Kansas City Health Department (KCHD). This incident provided the KCHD the first opportunity to operationalize its Incident Management System-based response plan. However, accompanying its implementation were unforeseen issues related to both internal and external communications. These issues and the lessons learned are discussed. PMID- 16210685 TI - Anthrax in New Jersey: a health education experience in bioterrorism response and preparedness. AB - The anthrax attack in 2001 created new challenges to health educators working on the response effort in New Jersey. Never before had there been a need for educating a group of people who had been exposed to a biological weapon. Coming on the heels of the catastrophic World Trade Center collapse on September 11, 2001, the New Jersey Department of Health and Senior Services was entrenched in the response to, and management of, the anthrax attack that placed a heavy emphasis on educating the postal workers of the United States Postal Service Trenton Processing and Distribution Center. This article provides an account of the preparation and delivery of educational materials and activities in the midst of a biological emergency, emphasizes the role health educators play in responding to bioterrorism events, and encourages health educators to become involved in bioterrorism preparedness efforts. PMID- 16210686 TI - Marketing physical activity: lessons learned from a statewide media campaign. AB - Steps taken to create, implement, and initially assess a statewide physical activity social marketing campaign targeted to 18-to 30-year-olds are presented. Included is a summary demonstration of the application of the associative group analysis in formative market research and message development. Initial postcampaign questionnaire (n = 363) results indicated that 39.1% of respondents had seen the television ad, of which 31.2% indicated they intended to be more active, and 62.5% of respondents had been exposed to either the television or outdoor media ads. Lessons learned through the social marketing process including media channel effectiveness, message development and assessment, and marketing firm relationships are provided. PMID- 16210687 TI - Physical activity among minority populations: what health promotion practitioners should know--a commentary. AB - Leisure time and moderate-level physical activity participation in the United States is low among the majority of the population. Minority populations are especially inactive and report having fewer opportunities and access to be physically active. Physical activity programs utilizing a strong health promotion framework (including needs assessment, program plan and design, program implementation, and evaluation) are limited, particularly among minority populations. The purpose of this article is to discuss the importance of physical activity, briefly review and describe current literature, identify the need for physical activity programs among minority populations, and relate the concepts of a health promotion framework to physical activity programs that are usable among practitioners. The article also provides health promotion practitioners with various resources for improving physical activity programs. PMID- 16210688 TI - Development of an ecological assessment tool for a workplace physical activity program standard. AB - This project was undertaken to develop a Workplace Physical Activity Assessment Tool to evaluate workplace physical activity programs based on a standard of best practices indicated in the literature and by key stakeholders. The development of the Assessment Tool was structured on an ecological model for health and physical activity promotion and an occupational health and safety audit instrument. The project included a review and synthesis of pertinent literature, expert and stakeholder reviews, interrater reliability appraisals, and workplace consultations over three distinct study phases. The Assessment Tool was received positively by the expert reviewers (i.e., academics, policy makers, and practitioners) and revealed generally high interrater reliability. The Workplace Physical Activity Assessment Tool can be used to plan, implement, and evaluate physical activity programs in the workplace. Recommendations for future research are identified. PMID- 16210689 TI - Online discussions with pregnant and parenting adolescents: perspectives and possibilities. AB - The Internet is an innovative strategy to increase public participation. It is important to include pregnant and parenting teens' perspectives when planning programs to meet their needs. This qualitative study explored online discussions as a strategy to enhance participation by this population. Findings showed that online communication was preferred over face-to-face group discussions. Being anonymous online encouraged open and honest feedback. Participants experienced various forms of social support, however, there was an overall lack of teen involvement online. Strategies to engage adolescents in online discussions and reduce barriers are discussed. Strategies included the use of teen moderators, home computer access, technical support, and engagement in naturally flowing online discussions to meet social support needs. Blending researchers' with teens' needs for social support in an online environment is encouraged. With careful planning and design, online communications can result in mutual benefits for researchers, service providers, and pregnant and parenting adolescents. PMID- 16210690 TI - Development, adaptation, and implementation of a cardiovascular health program for Alaska native women. AB - Southcentral Foundation's Traditions of the Heart program is an innovative cardiovascular disease prevention program for women designed to build on the strengths of the Alaska Native culture as a way to support and encourage positive lifestyle behaviors that focus on healthy eating, active living, stress management, and tobacco cessation. After conducting assessments of existing intervention programs and formative data collection, we adapted two existing programs, Native Nutrition Circles and A New Leaf... Choices for Healthy Living, to develop the Traditions of the Heart program. We implemented and evaluated a pilot intervention study to determine the program's acceptance among Alaska Native women. We used the evaluation results to further refine our study protocol. This article describes the adaptation of these programs to the cultural needs and strengths of Alaska Native women and the results of the formative evaluation used to improve the program design. The complete pilot study outcomes will be published separately. PMID- 16210691 TI - Managing fear in public health campaigns: a theory-based formative evaluation process. AB - The HIV/AIDS infection rate of Ethiopia is one of the world's highest. Prevention campaigns should systematically incorporate and respond to at-risk population's existing beliefs, emotions, and perceived barriers in the message design process to effectively promote behavior change. However, guidelines for conducting formative evaluation that are grounded in proven risk communication theory and empirical data analysis techniques are hard to find. This article provides a five step formative evaluation process that translates theory and research for developing effective messages for behavior change. Guided by the extended parallel process model, the five-step process helps message designers manage public's fear surrounding issues such as HIV/AIDS. An entertainment education project that used the process to design HIV/AIDS prevention messages for Ethiopian urban youth is reported. Data were collected in five urban regions of Ethiopia and analyzed according to the process to develop key messages for a 26 week radio soap opera. PMID- 16210692 TI - Oxygen and glucocorticoids modulate alphaENaC mRNA translation in fetal distal lung epithelium. AB - Glucocorticoid hormones play an important role in fetal lung maturation. It is unknown how they interact with changes in O2 tension, which play an important role in converting the lung from a fluid-secreting to a fluid-absorbing organ at birth. Airspace fluid absorption arises from active transepithelial Na+ transport with the amiloride-sensitive epithelial Na channel (ENaC), consisting of alpha, beta, and gamma subunits, representing the rate-limiting step under nonpathologic conditions. We investigated the individual and combined effects of dexamethasone (DEX) and PO2 on alphaENaC mRNA levels, rate of alphaENaC protein synthesis, and amiloride-sensitive short-circuit current in primary cultures of rat fetal distal lung epithelial cells. DEX significantly induced alphaENaC mRNA in fetal (3%) and postnatal (21%) O2, but increases in alphaENaC protein synthesis and function occurred only when epithelia were grown under a postnatal PO2. Sucrose density gradient analyses showed that DEX treatment of cells cultured at 3% O2 decreased the association of alphaENaC mRNA with large polysomes and enhanced the association with small polysomes. Conversely, incubation of DEX-treated cells in 21% O2 restored alphaENaC mRNA association with large polysomes. No significant changes were seen in the overall polyribosome profiles or in the distribution of mRNAs encoding beta and gamma subunits of ENaC or cytokeratin 18, indicating specific modulation of alphaENaC mRNA translation. These data suggest that postnatal O2 exposure may be important for efficient translation of the alphaENaC mRNA. PMID- 16210693 TI - Interleukin-13 and interleukin-4 induce vascular endothelial growth factor release from airway smooth muscle cells: role of vascular endothelial growth factor genotype. AB - Th2 cytokines induce the release of vascular endothelial growth factor (VEGF) from cultured human airway smooth muscle cells. The objective of this study was to examine the mechanistic basis for IL-4- and IL-13-induced VEGF release and to determine whether genetic differences are responsible for donor-to-donor variability in VEGF release. We measured VEGF mRNA expression by real-time PCR, mRNA stability using actinomycin D, and promoter activity with a VEGF-promoter luciferase reporter construct. We measured IL-4- and IL-13-induced VEGF release in cells from 21 donors by ELISA, genotyped the cells for common single nucleotide polymorphisms in the IL-4R alpha (Ile50Val, Ser478Pro, and Gln551Arg) and VEGF (-460T/C, -160C/T, -152G/A, +405C/G and +936 C/T) genes, and stratified the data by IL-4R alpha and VEGF genotype. IL-4 and IL-13 increased VEGF release and VEGF mRNA expression. IL-4 also increased mRNA stability but did not affect VEGF promoter activity. There was marked donor-to-donor variability in VEGF release from smooth muscle cells. The presence of Val50, Pro478/Arg551, or the Val50/Pro478/Arg551 IL-4R alpha haplotype had little effect on VEGF release. VEGF genotype at +405 or +936 alone had no effect on VEGF release, whereas cells bearing at least one -460C/-152A/+405G VEGF allele had lower release of VEGF in response to IL-13 or IL-4 than cells with other genotypes. Our data suggest that IL-4 and IL-13 mediate their effects on VEGF expression post-transcriptionally and indicate that polymorphisms in the VEGF, but not the IL-4R alpha, gene affect VEGF release from smooth muscle cells. PMID- 16210695 TI - Contribution of TNF-alpha converting enzyme and proteinase-3 to TNF-alpha processing in human alveolar macrophages. AB - Membrane-associated TNF-alpha cleavage is required to yield the 17.5-kD soluble product. This process is poorly understood in human cells, and no studies have related this process to the alveolar macrophage (AM). TNF-alpha-converting enzyme (TACE) is known to cleave TNF at the Ala-76-Val-77 site. We have evaluated the expression, regulation, and catalytic function of TACE in healthy human AMs. TACE was detected on the surface of AMs using flow cytometry. TACE protein can be upregulated by LPS (P = 0.036) and IFN-gamma. LPS-induced expression is downregulated by IL-10 (P = 0.04) and TNF-alpha. TACE regulation was observed at the mRNA level. TACE catalytic activity as assessed by cleavage of glutathione S transferase-proTNF fusion protein correlates significantly with TACE protein expression (P = 0.04). However, cleavage and soluble TNF-alpha release by AMs was inhibited by matrix metalloproteinase and serine protease inhibitors, suggesting a role for a serine protease in this process. We confirmed the presence of proteinase-3 (PR-3) on the AM surface that was functionally capable of TNF cleavage. PR-3 mRNA expression was not found in AMs. However, we determined that PR-3 from neutrophil supernatants could bind to the AM membrane, suggesting that AM-derived PR-3 is from an exogenous source, which is important in the context of inflammation. PMID- 16210694 TI - Nuclear factor-kappaB affects tumor progression in a mouse model of malignant pleural effusion. AB - We developed a novel mouse model of malignant pleural effusion (MPE) by injecting Lewis lung cancer (LLC) cells directly into the pleural space of syngeneic C57B/6 mice. The pleural effusions in this model share common cellular and biochemical features with human MPEs. Implantation and growth of pleural tumors triggers a host inflammatory response characterized by a mixed inflammatory cell influx into the pleural fluid. LLC cells exhibited high basal nuclear factor (NF)-kappaB activity in vitro and in vivo, which we used to drive expression of a NF-kappaB dependent green fluorescent protein-firefly luciferase fusion reporter construct. NF-kappaB-dependent reporter expression allowed intravital tracing of pleural tumors. Inhibition of NF-kappaB in LLC cells did not affect cell viability in culture; however, injection of LLC cells expressing a dominant NF-kappaB inhibitor resulted in decreased tumor burden, decreased pleural effusion volume, and decreased pleural effusion TNF-alpha levels. These studies indicate that tumor NF-kappaB activity regulates pleural tumor progression. This reproducible model of MPE can be used to further study the influence of specific host and tumor factors on the pathogenesis of MPE and evaluate new therapeutic strategies. PMID- 16210696 TI - Rhinovirus induces airway epithelial gene expression through double-stranded RNA and IFN-dependent pathways. AB - Rhinovirus (RV) infection is the major cause of common colds and of asthma exacerbations. Because the epithelial cell layer is the primary target of RV infection, we hypothesize that RV-induced airway disease is associated with the perturbation of airway epithelial gene expression. In this study, well differentiated primary human airway epithelial cells were infected with either RV16 (major group) or RV1B (minor group). Transcriptional gene profiles from RV infected and mock-infected control cells were analyzed by Affymetrix Genechip, and changes of the gene expression were confirmed by real-time RT-PCR analysis. At 24 h after infection, 48 genes induced by both viruses were identified. Most of these genes are related to the IFN pathway, and have been documented to have antiviral functions. Indeed, a significant stimulation of IFN-beta secretion was detected after RV16 infection. Neutralizing antibody specific to IFN-beta and a specific inhibitor of the Janus kinase pathway both significantly blocked the induction of RV-inducible genes. Further studies demonstrated that 2-aminopurine, a specific inhibitor double-stranded RNA-dependent protein kinase, could block both IFN-beta production and RV-induced gene expression. Thus, IFN-beta-dependent pathway is a part of the double-stranded RNA-initiated pathway that is responsible for RV-induced gene expression. Consistent with its indispensable role in the induction of antiviral genes, deactivation of this signaling pathway significantly enhanced viral production. Because increase of viral yield is associated with the severity of RV-induced airway illness, the discovery of an epithelial antiviral signaling pathway in this study will contribute to our understanding of the pathogenesis of RV-induced colds and asthma exacerbations. PMID- 16210697 TI - Do low doses of folic acid result in maximum lowering of homocysteine? PMID- 16210698 TI - Choline, homocysteine, and pregnancy. PMID- 16210699 TI - Concept of a nutritious food: toward a nutrient density score. AB - The American diet is said to be increasingly energy-rich but nutrient-poor. To help improve the nutrient-to-energy ratio, the 2005 Dietary Guidelines for Americans recommend that consumers replace some foods in their diets with more nutrient-dense options. Such dietary guidance presupposes the existence of a nutrient density standard. However, a review of the literature shows that the concept of a nutritious food is not based on any consistent standards or criteria. In many cases, healthful foods are defined by the absence of problematic ingredients-fat, sugar, and sodium-rather than by the presence of any beneficial nutrients they might contain. Past attempts to quantify the nutrient density of foods have been based on a variety of calories-to-nutrient scores, nutrients-per-calorie indexes, and nutrient-to-nutrient ratios. The naturally nutrient rich (NNR) score, which is based on mean percentage daily values (DVs) for 14 nutrients in 2000 kcal food, can be used to assign nutrient density values to foods within and across food groups. Use of the NNR score allows consumers to identify and select nutrient-dense foods while permitting some flexibility where the discretionary calories are concerned. This approach has implications for food labeling, nutritional policy making, and consumer education. The Food and Drug Administration has considered approving nutrient claims based on the ratio of a beneficial nutrient to the food's energy content, as opposed to a specified minimum amount of a nutrient per serving size. Given the current dietary trends, the nutrient density approach can be a valuable tool for nutrition education and dietary guidance. PMID- 16210700 TI - Age at menarche and adult BMI in the Aberdeen children of the 1950s cohort study. AB - BACKGROUND: Few studies have examined whether the inverse association of age at menarche with adult body mass index (BMI) is due to the tendency of BMI to track between childhood and adult life, with childhood BMI playing a causal role in determining age at menarche. OBJECTIVE: The objective was to investigate whether the association of younger age at menarche with a high BMI and increased likelihood of obesity in middle age is due to confounding with early childhood BMI. DESIGN: In a historical cohort of 3743 Scottish females born between 1950 and 1955, height and weight were measured in early childhood, and age at menarche and height and weight in middle age were obtained by questionnaire. RESULTS: The age-adjusted change in mean adult BMI per additional year of age at menarche was 0.64 (95% CI: -0.78, -0.50). Adjustment for childhood BMI measured between 4 and 6 y reduced this value to -0.57 (-0.71, -0.43). Adjustment for childhood and adult social class, parity, smoking, and alcohol intake had little effect. The odds ratio for being obese compared with not being obese in adulthood was 0.82 (0.76, 0.86) per 1-y increase in age at menarche and was unchanged by adjustment for childhood BMI and other covariates. CONCLUSIONS: The inverse association of age at menarche with BMI and obesity in middle age is not explained by confounding by early childhood BMI. Instead, age at menarche may simply be a proxy marker for the pace of sexual maturation, which itself leads to differences in adiposity (and BMI) in the peripubertal period that track into adult life. PMID- 16210701 TI - Relation of weight maintenance and dietary restraint to peroxisome proliferator activated receptor gamma2, glucocorticoid receptor, and ciliary neurotrophic factor polymorphisms. AB - BACKGROUND: Genetic variation in the peroxisome proliferator-activated receptor gamma2 (PPARgamma2), glucocorticoid receptor (GRL), and ciliary neurotrophic factor (CNTF) genes may play a role in the etiology of obesity. OBJECTIVE: We examined biological, psychological, and genetic determinants associated with weight maintenance (WM) after weight loss. DESIGN: Subjects (n = 120) followed a 6-wk diet and then a 1-y period of WM. Body weight (BW), body composition, leptin concentration, attitude toward eating (measured with the Three-Factor Eating Questionnaire), physical activity, and the polymorphisms of the PPARgamma2, GRL, and CNTF genes were measured. RESULTS: BW loss was 7.0 +/- 3.1 kg. After 1 y, 21 subjects showed successful WM (<10% regain); 99 were unsuccessful (> or =10% regain). Compared with unsuccessful subjects, successful subjects had a higher increase in dietary restraint over time (4.8 +/- 5.0 and 1.8 +/- 3.9, respectively; P < 0.01) but significantly less sensation of general hunger (-4.0 +/- 4.9 and -1.2 +/- 2.7, respectively; P < 0.05). Successful subjects had a significantly different frequency distribution for the PPARgamma2 (P = 0.05) and GRL (P < 0.05) genes than did unsuccessful subjects. The more successful genotypes showed a higher baseline body mass index and waist circumference (PPARgamma2), a greater decrease in disinhibition of dietary restraint (GRL), and less sensation of hunger (GRL). The G/G genotype (GRL) was an independent predictor of successful WM. CONCLUSION: The different genotypes of the PPARgamma2 and GRL genes contribute to WM, either directly (GRL) or indirectly (PPARgamma2 and GRL) via baseline body mass index and waist circumference, and to changes in Three-Factor Eating Questionnaire scores. PMID- 16210702 TI - Plasma palmitoleic acid content and obesity in children. AB - BACKGROUND: Palmitoleic acid (16:1n-7) is a product of endogenous lipogenesis. In human obesity, 16:1n-7 is reported to correlate with indexes of adiposity and insulin concentrations. OBJECTIVE: We investigated the relation between adiposity, especially in the abdominal region, and plasma monounsaturated fatty acid (MUFA) profiles in obese children. DESIGN: A case-control study was performed. The study subjects were 59 obese children (x +/- SD age: 11.8 +/- 3.8 y) and 53 age- and sex-matched healthy, nonobese children (aged 12.5 +/- 0.5 y). The study's variables included anthropometric measurements, serum lipids, leptin, and fatty acid composition in plasma. RESULTS: MUFA profiles of obese subjects showed a significantly higher content of 16:1n-7, 18:1n-9, and 20:1n-9 and significantly higher stearoyl-CoA desaturase (SCD) activity (ratio of 16:1n-7 to 16:0) than in nonobese controls. In a multiple regression analysis, percentage body fat, waist-to-height ratio, and waist-to-hip ratio (WHR) were significant determinants of 16:1n-7 content. SCD activity had a positive, significant correlation with leptin. However, in a multiple regression analysis that included percentage body fat, WHR, and leptin as independent determinants, WHR was the only determinant of SCD activity. CONCLUSIONS: Plasma 16:1n-7 content has a significant relation with abdominal adiposity in obese children. This change in the MUFA profile may be caused by activation of SCD that is not sufficiently suppressed by leptin. Endogenous lipogenesis may be an important factor in the pathogenesis of obesity in children. PMID- 16210703 TI - Arterial endothelial dysfunction in baboons fed a high-cholesterol, high-fat diet. AB - BACKGROUND: Endothelial dysfunction signals the initiation and progression of atherosclerosis. Elevated LDL-cholesterol concentrations have been suggested to induce endothelial dysfunction, but direct in vivo evidence for the relation is still lacking. OBJECTIVE: We examined the hypothesis that a high-cholesterol, high-fat (HCHF) diet can directly cause endothelial dysfunction in vivo. DESIGN: We measured inflammatory and endothelial dysfunctional markers in circulating blood and directly in endothelial cells, which were collected by femoral artery biopsies, in 10 baboons before and after a 7-wk HCHF dietary challenge. RESULTS: We found that the HCHF diet induced a high inflammatory status, as indicated by increased concentrations of interleukin 6, tumor necrosis factor alpha (TNF alpha), and monocyte chemoattractant protein 1. Although the concentrations of endothelial dysfunctional markers, such as soluble vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1, were not increased by the HCHF diet, membrane-bound VCAM-1 and membrane-bound E-selectin on endothelial cells were highly increased after 7 wk of the HCHF diet (P < 0.01). In contrast, the concentrations of endothelial nitric oxide synthase in endothelial cells were significantly reduced by the 7-wk HCHF diet (P < 0.01). Furthermore, the dietary challenge attenuated endothelial cell responses to TNF-alpha, lipopolysaccharide, native LDL cholesterol, and oxidized LDL-cholesterol stimulation. CONCLUSIONS: Our results show that an HCHF diet can directly induce inflammation and endothelial dysfunction. Prior in vivo exposure to an HCHF diet attenuates the in vitro responses of endothelial cells to atherogenic risk factors. This preconditioning phenomenon may have significant clinical relevance. PMID- 16210704 TI - Association of the 276G->T polymorphism of the adiponectin gene with cardiovascular disease risk factors in nondiabetic Koreans. AB - BACKGROUND: The adiponectin gene is known to modulate adiponectin concentrations and diabetes mellitus development. OBJECTIVE: We assessed whether adiponectin gene variants contribute to circulating adiponectin, insulin resistance (IR), or cardiovascular disease risk factors. DESIGN: Nondiabetic subjects [n = 902; x +/- SE age: 42.5 +/- 0.53 y; body mass index (BMI; in kg/m2): 24.7 +/- 0.11] were genotyped for 2 single-nucleotide polymorphisms (SNPs), 45T-->G and 276G-->T. RESULTS: After adjustment for age, sex, and BMI, subjects with the G allele for the SNP 276 had significantly higher concentrations of triacylglycerol and small dense LDL (sdLDL) and smaller LDL particle size than did T/T subjects. G/G subjects at SNP 276 had significantly lower plasma adiponectin and higher homeostasis model assessment (HOMA) of IR and urinary prostaglandin F2alpha than did T/T subjects. In the SNP 45-276 haplotype test, we also observed that subjects with the X/X haplotype had significantly higher plasma adiponectin after adjustment than did TG/TG or TG/X haplotype subjects. In the highest BMI group (BMI > or = 26), T/T subjects had lower HOMA-IR (P = 0.011) and higher plasma adiponectin (P = 0.026) at SNP 276 than did G/G or G/T subjects. These patterns were also seen for adiponectin in haplotype groups. However, no significant genotype effect for SNP 45T-->G was observed. CONCLUSIONS: The 276G-->T polymorphism of the adiponectin gene modulates circulating adiponectin and IR, particularly in obese states. G allele carriers also have higher oxidative stress, higher sdLDL concentrations, and smaller LDL particle size. Therefore, the presence of the G allele in the adiponectin gene at SNP 276 could be a significant contributor to higher cardiovascular disease risk in Koreans, independent of common environmental factors. PMID- 16210705 TI - 8-Isoprostane F2alpha excretion is reduced in women by increased vegetable and fruit intake. AB - BACKGROUND: Health benefits associated with diets rich in vegetables and fruit (VF) are often attributed to the antioxidant activity of their constituent phytochemicals. However, in vivo evidence that VF actually reduce markers of oxidative stress is limited. OBJECTIVE: An 8-wk dietary intervention was conducted to test the hypothesis that increased VF consumption decreases oxidative stress. Urinary excretion of 8-isoprostane F2alpha (8-iso-PGF2alpha) was used as an index of whole-body lipid peroxidation. DESIGN: The diets evaluated had comparable amounts of all macronutrients but varied in their content of VF. After a 2-wk low-VF (3.0 servings/d) run-in diet, 246 women were randomly assigned to receive either 3.6 (low) or 9.2 (high) servings VF/d. The low-VF group was switched to the high-VF diet during the final 2 wk of the study. Blood and first-void urine specimens were obtained at baseline and at 2-wk intervals thereafter. RESULTS: The run-in diet reduced 8-iso-PGF2alpha concentrations by 33% (P < 0.0001). The excretion of 8-iso-PGF2alpha with the low VF diet remained the same as that with the run-in diet, whereas urinary concentrations of 8-iso-PGF2alpha were further reduced (P < 0.01) by the high-VF diet, either fed throughout the study or when the diet was switched from low to high VF (P = 0.05). The greatest reductions in 8-iso-PGF2alpha were observed in subjects in the highest quartile of baseline concentrations of 8-iso-PGF2alpha. CONCLUSIONS: A significant reduction in the excretion of 8-iso-PGF2alpha was induced by the run-in diet and the high-VF diet. The degree of reduction was related to the subject's baseline urinary concentration of 8-iso-PGF2alpha. PMID- 16210706 TI - Geriatric Nutritional Risk Index: a new index for evaluating at-risk elderly medical patients. AB - BACKGROUND: Patients at risk of malnutrition and related morbidity and mortality can be identified with the Nutritional Risk Index (NRI). However, this index remains limited for elderly patients because of difficulties in establishing their normal weight. OBJECTIVE: Therefore, we replaced the usual weight in this formula by ideal weight according to the Lorentz formula (WLo), creating a new index called the Geriatric Nutritional Risk Index (GNRI). DESIGN: First, a prospective study enrolled 181 hospitalized elderly patients. Nutritional status [albumin, prealbumin, and body mass index (BMI)] and GNRI were assessed. GNRI correlated with a severity score taking into account complications (bedsores or infections) and 6-mo mortality. Second, the GNRI was measured prospectively in 2474 patients admitted to a geriatric rehabilitation care unit over a 3-y period. RESULTS: The severity score correlated with albumin and GNRI but not with BMI or weight:WLo. Risk of mortality (odds ratio) and risk of complications were, respectively, 29 (95% CI: 5.2, 161.4) and 4.4 (95% CI: 1.3, 14.9) for major nutrition-related risk (GNRI: <82), 6.6 (95% CI: 1.3, 33.0), 4.9 (95% CI: 1.9, 12.5) for moderate nutrition-related risk (GNRI: 82 to <92), and 5.6 (95% CI: 1.2, 26.6) and 3.3 (95% CI: 1.4, 8.0) for a low nutrition-related risk (GNRI: 92 to < or =98). Accordingly, 12.2%, 31.4%, 29.4%, and 27.0% of the 2474 patients had major, moderate, low, and no nutrition-related risk, respectively. CONCLUSION: GNRI is a simple and accurate tool for predicting the risk of morbidity and mortality in hospitalized elderly patients and should be recorded systematically on admission. PMID- 16210707 TI - Poor nutritional habits are predictors of poor outcome in very old hospitalized patients. AB - BACKGROUND: Malnutrition is prevalent in elderly populations. Recommended methods of nutritional screening are often too complicated and time-consuming for routine application in frail, very old, hospitalized patients. OBJECTIVE: Our aims were to identify risk factors for development of malnutrition in very old hospitalized patients and to evaluate the total Mini Nutritional Assessment (MNA) score and MNA subscores as predictors of in-hospital and long-term mortality. DESIGN: A prospective cohort study of patients aged > or =75 y was conducted in a geriatric hospital. Assessment included demographic, clinical, and laboratory data and cognitive, functional, and nutritional status. Follow-up was conducted for < or =2.7 y. RESULTS: Of the 414 patients studied, only 73 (17.6%) were well nourished. Low serum albumin and phosphorus concentrations, dementia, and cerebrovascular accident (CVA) were significant risk factors for malnutrition. Survival was significantly lower in malnourished patients and patients at risk of malnutrition than in well-nourished patients (P < 0.0001). Low MNA-3 subscores (dietary habits) were significantly correlated with laboratory indexes of malnutrition and were significantly lower in patients with infections, malignancy, pressure ulcers, dementia, recent orthopedic surgery, and CVA. Multivariate analysis showed that a low MNA-3 score was an independent predictor of mortality; scores <7.5 increased the risk of death 2.05-fold. CONCLUSIONS: The prevalence of malnutrition was high in elderly hospitalized patients. Dietary habits were significant predictors of poor hospitalization outcome. A questionnaire on dietary habits can serve as a useful tool in assessing nutritional status and prognosis in elderly patients. PMID- 16210708 TI - Protein kinetic differences between children with edematous and nonedematous severe childhood undernutrition in the fed and postabsorptive states. AB - BACKGROUND: Pathogenic factors that cause a child to develop the edematous instead of the nonedematous form of severe childhood undernutrition (SCU) during food deprivation are not clear. It was hypothesized that, in edematous but not nonedematous SCU, impaired protein breakdown leading to inadequate amino acids for maintenance of important organ systems was a factor. OBJECTIVE: We measured protein kinetics in children with edematous and nonedematous SCU. DESIGN: Endogenous leucine flux, an index of whole-body protein breakdown rate, was determined in 4 groups of children with edematous or nonedematous SCU in the malnourished and recovered states. Two groups were studied in the postabsorptive state, and 2 groups were studied in the fed state. RESULTS: In the postabsorptive state, leucine flux was slower (P < 0.01) in the edematous group than in the nonedematous group in the malnourished state, but in the recovered state, it was faster (P < 0.05) in the children who previously had edematous SCU. When compared with the malnourished state value, leucine flux at recovery doubled in the group that previously had edematous SCU, but it did not change in the other group. In the fed state, leucine flux was slower (P < 0.01) in the edematous group than in the nonedematous group in the malnourished state but not in the recovered state. In the recovered state, enteral leucine extraction by splanchnic tissues trended higher in the group that previously had edematous SCU than in the nonedematous group. CONCLUSION: These findings indicate different protein breakdown responses to food deprivation between children with edematous and nonedematous SCU and inherent differences in protein metabolism when they have recovered. PMID- 16210709 TI - Inflammation is associated with increased energy expenditure in patients with chronic kidney disease. AB - BACKGROUND: Inflammation, a clinical condition observed in patients with chronic kidney disease (CKD), may be related to increased resting energy expenditure (REE). OBJECTIVES: The main objective was to investigate the relation between inflammation and REE in patients with CKD who are not undergoing dialysis. We also aimed to analyze whether a decrease in C-reactive protein (CRP) would result in a reduction in REE. DESIGN: This study enrolled 132 patients with CKD who were not undergoing dialysis, who had creatinine clearance from 5 to 65 mL.min( 1).1.73 m(-2), and who were 53.6 +/- 16 y old; 82 (62.1%) were men. Twenty-nine patients had clinical signs of infection. REE was measured by using indirect calorimetry, and inflammation was evaluated by using high-sensitivity CRP measurement. Patients were divided according to tertiles of CRP with the following intertertile ranges: first tertile, CRP < or = 0.14 mg/dL (n = 43); second tertile, CRP 0.15-0.59 mg/dL (n = 46); and third tertile, CRP > or = 0.60 mg/dL (n = 43). REE was measured before and after treatment in 10 patients who had inflammation or infection. RESULTS: After adjustment for age, sex, and lean body mass, the REE of the third (1395 kcal/d; P = 0.02) and second (1355 kcal/d; P = 0.04) tertiles was significantly higher than that of the first tertile (1286 kcal/d). In the multiple linear regression analysis (n = 132), the independent determinants of REE were lean body mass, CRP, and age (R2 = 0.55). After treatment of infection in a subgroup of 10 patients, it was observed that a significant reduction in CRP concentration was accompanied by a significant reduction of 174 +/- 165 kcal that accounted for 13% of the initial REE. CONCLUSION: This study showed that inflammation is associated with increased REE in patients with CKD. PMID- 16210710 TI - Dose-dependent effects of folic acid on blood concentrations of homocysteine: a meta-analysis of the randomized trials. AB - BACKGROUND: Dietary supplementation with B vitamins that lower blood homocysteine concentrations is expected to reduce cardiovascular disease risk, but there has been uncertainty about the optimum regimen to use for this purpose. OBJECTIVE: The objectives were to ascertain the lowest dose of folic acid associated with the maximum reduction in homocysteine concentrations and to determine the additional relevance of vitamins B-12 and B-6. DESIGN: A meta-analysis of 25 randomized controlled trials involving individual data on 2596 subjects assessed the effect on plasma homocysteine concentrations of different doses of folic acid and of the addition of vitamins B-12 and B-6. RESULTS: The proportional reductions in plasma homocysteine concentrations produced by folic acid were greater at higher homocysteine (P < 0.001) and lower folate (P < 0.001) pretreatment concentrations; they were also greater in women than in men (P < 0.001). After standardization for sex and to pretreatment plasma concentrations of 12 micromol homocysteine/L and 12 nmol folate/L, daily doses of 0.2, 0.4, 0.8, 2.0, and 5.0 mg folic acid were associated with reductions in homocysteine of 13% (95% CI: 10%, 16%), 20% (17%, 22%), 23% (21%, 26%), 23% (20%, 26%), and 25% (22%, 28%), respectively. Vitamin B-12 (x: 0.4 mg/d) produced 7% (95% CI: 4%, 9%) further reduction in homocysteine concentrations, but vitamin B-6 had no significant effect. CONCLUSIONS: Daily doses of > or =0.8 mg folic acid are typically required to achieve the maximal reduction in plasma homocysteine concentrations produced by folic acid supplementation. Doses of 0.2 and 0.4 mg are associated with 60% and 90%, respectively, of this maximal effect. PMID- 16210712 TI - Spinach or carrots can supply significant amounts of vitamin A as assessed by feeding with intrinsically deuterated vegetables. AB - BACKGROUND: The vitamin A value of spinach and carrots needs to be measured directly. OBJECTIVE: The objective was to determine the vitamin A value of intrinsically labeled dietary spinach and carrots in humans. DESIGN: Spinach and carrots were intrinsically labeled by growing these plants in 25 atom% 2H2O nutrient solution. Growth in this medium yielded a range of trans beta-carotene (tbeta-carotene) isotopomers with a peak enrichment at molecular mass plus 10 mass units. Seven men with a mean (+/-SD) age of 59.0 +/- 6.3 y and a body mass index (in kg/m2) of 25.7 +/- 1.5 consumed pureed spinach (300 g, 20.8 micromol tbeta-carotene equivalents) or carrots (100 g, 19.2 micromol tbeta-carotene equivalents) with a standardized liquid diet (no extra fiber) in random order 4 mo apart. Seven women with a mean (+/-SD) age of 55.5 +/- 6.3 y and a body mass index of 26.4 +/- 4.2 consumed pureed spinach only (300 g, 20.0 micromol tbeta carotene equivalents). A reference dose of [13C8]retinyl acetate (8.9 micromol) in oil was given to each subject 1 wk after each vegetable dose. Blood samples were collected over 35 d. RESULTS: Areas under the curve for total labeled serum beta-carotene responses were 42.4 +/- 8.5 nmol.d per micromol spinach beta carotene and 119.8 +/- 23.0 nmol.d per micromol carrot beta-carotene (P < 0.01). Compared with the [13C8]retinyl acetate reference dose, spinach tbeta-carotene conversion to retinol was 20.9 +/- 9.0 to 1 (range: 10.0-46.5 to 1) and carrot tbeta-carotene conversion to retinol was 14.8 +/- 6.5 to 1 (range: 7.7-24.5 to 1) by weight. CONCLUSIONS: Spinach and carrots can provide a significant amount of vitamin A even though the amount is not as great as previously proposed. Food matrices greatly affect the bioavailability of plant carotenoids, their efficiency of conversion to vitamin A, or both. PMID- 16210711 TI - Diet and genetic factors associated with iron status in middle-aged women. AB - BACKGROUND: Gene mutations associated with iron overload have been identified. How food and nutrient intakes affect iron status in persons who may be at risk of iron overload because their genetic status is unknown. OBJECTIVE: The objective was to determine the relation between food and nutrient intakes, HFE genotype, and iron status. Foods and nutrients associated with iron stores, with adjustment for gene mutations associated with hemochromatosis, were explored. DESIGN: A prospective cohort of women aged 35-69 y (the UK Women's Cohort Study) provided information on diet through a questionnaire and food diary; 6779 women in the cohort provided cheek cell samples, blood samples, or both, which were genotyped for C282Y and H63D mutations, and 2489 women also had their iron status assessed. Relations between serum ferritin and iron intake were investigated by using multiple linear regression, with adjustment for potential confounders. RESULTS: The strongest dietary association with serum ferritin concentration was a positive association with heme iron and not with nonheme or total iron. Weaker positive associations were seen with red and white meat, and negative associations were seen with total energy and white and brown whole-meal bread, independent of genotype and other potential confounders. The effect of genotype on ferritin concentrations primarily occurred after menopause, at which time a strong interaction between genotype and heme iron intake was observed. Other factors associated with serum ferritin concentrations were age, body mass index, blood donation, menopausal status, and HFE genotype. CONCLUSIONS: Postmenopausal women eating a diet rich in heme iron and who were C282Y homozygotes had the highest serum ferritin concentrations. PMID- 16210713 TI - Efficacy of twice-weekly multiple micronutrient supplementation for improving the hemoglobin and micronutrient status of anemic adolescent schoolgirls in Bangladesh. AB - BACKGROUND: Although iron deficiency is a major cause of anemia, other micronutrient deficiencies may also play a role. OBJECTIVE: We examined whether multiple micronutrient supplementation is more efficacious than is supplementation with iron and folic acid alone for improving the hemoglobin and iron status of anemic adolescent girls in Bangladesh. DESIGN: Anemic (hemoglobin < 12.0 g/dL) girls (n = 197) aged 14-18 y from rural schools in Dhaka District were entered into a randomized double-blind trial and received twice-weekly supplements of iron and folic acid (IFA group) or multiple micronutrients (15 micronutrients, including iron and folic acid; MMN group) for 12 wk. RESULTS: At recruitment, the characteristics of the girls in the 2 groups were not significantly different, except for family size and body mass index. At the end of the study, although both groups benefited significantly from supplementation, mean changes in hemoglobin and serum ferritin concentrations were not significantly different between groups. Compared with the IFA group, girls in the MMN group had significantly greater increases in mean serum vitamin A, plasma vitamin C, red blood cell folic acid, and riboflavin concentrations (assessed as erythrocyte glutathione reductase activation coefficient). After 12 wk of supplementation, only the prevalence of vitamins A and C and riboflavin deficiencies decreased more significantly in the MMN group than in the IFA group. CONCLUSIONS: Twice-weekly MMN supplementation for 12 wk significantly improved the status of the micronutrients assessed but was not more efficacious than was supplementation with iron and folic acid alone in improving the hematologic status of anemic adolescent girls. More frequent doses may be needed to achieve full benefit. PMID- 16210714 TI - Choline and homocysteine interrelations in umbilical cord and maternal plasma at delivery. AB - BACKGROUND: Little is known about the interactions between choline and folate and homocysteine metabolism during pregnancy despite the facts that pregnancy places considerable stress on maternal folate and choline stores and that choline is a critical nutrient for the fetus. Choline, via betaine, is an important folate independent source of methyl groups for remethylating homocysteine in liver. OBJECTIVES: Our aims were to examine the intermediates of choline oxidation in maternal and umbilical cord plasma and to determine the relations between this pathway and folate-dependent homocysteine remethylation. DESIGN: Blood samples were taken from 201 pregnant women and, at delivery, from the umbilical cord veins of their healthy, full-term infants. The blood samples were analyzed for plasma free choline, betaine, dimethylglycine, folate, vitamin B-12, total homocysteine (tHcy), and creatinine concentrations. RESULTS: Choline concentrations in umbilical cord plasma were approximately 3 times those in maternal plasma (geometric x: 36.6 and 12.3 micromol/L, respectively; P < 0.0001). Betaine and dimethylglycine concentrations were also significantly higher in umbilical cord than in maternal plasma. Choline was positively associated with tHcy (r = 0.34, P < 0.0001), betaine (r = 0.58, P < 0.0001), and dimethylglycine (r = 0.30, P < 0.0001) in maternal blood. Much weaker relations were seen in the fetal circulation. In a multiple regression model, choline was a positive predictor of maternal tHcy, whereas vitamin B-12 and betaine were negative predictors. CONCLUSIONS: The positive association between maternal choline and tHcy during pregnancy suggests that the high fetal demand for choline stimulates de novo synthesis of choline in maternal liver, with a resultant increase in tHcy concentrations. If this is confirmed, it may be appropriate to provide choline supplements during pregnancy to prevent elevated tHcy concentrations. PMID- 16210715 TI - Effect of time of initiation and dose of prenatal iron and folic acid supplementation on iron and folate nutriture of Korean women during pregnancy. AB - BACKGROUND: In Korea, it is customary to prescribe iron and folic acid supplements to pregnant women after the 20th wk of gestation; however, little evidence exists to support this practice. OBJECTIVE: The objective was to determine the effects of time of initiation and dose of prenatal iron and folic acid supplementation on the iron and folate nutriture of Korean women during pregnancy. DESIGN: A total of 131 pregnant women were placed into 1 of 5 experimental groups, either the control group or 1 of 4 supplemented groups. The supplemented groups varied by time of initiation, which was either during the first trimester or at week 20 of gestation, and by dose of iron and folic acid supplements provided, which consisted of either 30 mg Fe plus 175 microg folic acid or 60 mg Fe plus 350 microg folic acid. All supplemented groups continued supplementation until delivery. RESULTS: Improvements in iron and folate nutriture were highly dependent on when the supplement program was initiated, but both supplement doses were equally effective. In contrast, the influence of folic acid supplementation on maternal folate status was not as pronounced as was the influence of iron supplementation on iron status. CONCLUSION: In pregnant Korean women, initiating iron and folic acid supplementation earlier during pregnancy may prevent the deterioration of iron and folate nutriture more than does increasing supplement doses in later stages of pregnancy. PMID- 16210716 TI - Sex differences in the associations of HIV disease characteristics and body composition in antiretroviral-naive persons. AB - BACKGROUND: Data on associations of body composition with HIV disease characteristics are limited. OBJECTIVE: We compared sex-specific associations between HIV disease characteristics and body composition in an racially ethnically diverse cohort of antiretroviral-naive patients. DESIGN: The study was a cross-sectional analysis of participants enrolled in a metabolic substudy of a multicenter trial. Regional fat was measured, and total body fat (TBF) was derived by using the Durnin-Womersley formula (DWF) and bioelectrical impedance analysis (BIA). Body cell mass (BCM) was measured by BIA. RESULTS: Among 422 participants, 22% were women, 60% were African American, and 36% had prior AIDS defining illnesses. Mean (+/-SD) age was 38.2 +/- 9.6 y, CD4+ count was 215 +/- 184 cells/mm3, and HIV RNA log10 was 5.0 +/- 0.8 copies/mL. On multivariate analysis, women with AIDS-defining illness had significantly (P < 0.005) lower regional body fat and TBF (BIA: -9.5 kg; DWF: -7.3 kg) but nonsignificantly lower BCM (-1.3 kg) than did women without such illnesses, whereas men with AIDS defining illness had significantly (P < 0.005) lower BCM (-1.7 kg) but nonsignificantly lower TBF (BIA: -1.3 kg; DWF: -1.83 kg) than did men without such illnesses (P < 0.05 for sex differences in TBF). Significant negative associations of HIV RNA with BCM (-0.9 kg/log RNA; P = 0.03), TBF by BIA (-1.4 kg/log RNA; P = 0.05) and by DWF (-1.6 kg/log RNA; P = 0.01), and regional fat were observed in men only. CONCLUSIONS: The effect of prior AIDS illness on body fat differed significantly between the sexes: women with prior AIDS-defining illness had significantly less fat than did women without such illnesses. An independent effect of HIV viremia on BCM and fat was seen in men. These distinctions may be due to inherent biological differences between the sexes. PMID- 16210717 TI - Vitamin supplements, socioeconomic status, and morbidity events as predictors of wasting in HIV-infected women from Tanzania. AB - BACKGROUND: Wasting is a strong independent predictor of mortality in HIV infected persons. Vitamin supplements delay the disease progression, but their effect on wasting is not known. Data are lacking on the risk factors for wasting in African HIV-infected persons. OBJECTIVES: The objectives were to examine the effect of vitamin supplements on wasting in HIV-infected women and to assess the effects of sociodemographic characteristics, morbidity events, and immunologic progression on the risk of wasting. DESIGN: HIV-infected women (n = 1078) from Tanzania were randomly assigned to receive 1 of 4 daily oral regimens: multivitamins (B complex, C, and E), vitamin A plus beta-carotene, multivitamins that included vitamin A plus beta-carotene, or placebo. The endpoints of the study included first episodes of a midupper arm circumference <22 cm or a body mass index (BMI; in kg/m2) <18 and the incidence of weight loss episodes during a median 5.3 y of follow-up. RESULTS: Multivitamins alone significantly reduced the risk of a first episode of a midupper arm circumference <22 cm (relative risk: 0.66; 95% CI: 0.47, 0.94; P = 0.02). In multivariate-adjusted Cox models, the woman's age, education level, and height were inversely related to the incidence of wasting. Episodes of diarrhea, nausea or vomiting, lower respiratory tract infections, oral ulcers, thrush, severe anemia, and low CD4+ cell counts were each significantly related to an increased risk of wasting. CONCLUSIONS: Vitamins C and E and the vitamin B complex have a protective effect on wasting in HIV infected women. Prevention of diarrhea, severe respiratory tract infections, and anemia are likely to decrease the burden of wasting. PMID- 16210718 TI - Homocysteine, vitamin B-12, and folic acid and the risk of cognitive decline in old age: the Leiden 85-Plus study. AB - BACKGROUND: High concentrations of homocysteine and low concentrations of vitamin B-12 and folic acid are frequently observed in subjects with dementia. OBJECTIVE: We assessed whether serum concentrations of homocysteine, vitamin B-12, or folic acid predict cognitive decline in old age. DESIGN: This was a prospective, population-based, longitudinal study of 599 subjects (Leiden 85-Plus Study, Netherlands). The subjects were administered a battery of cognitive tests (including the Mini Mental State Examination, the Stroop test, a letter digit coding test, and a word recall test) at 85 y of age and yearly thereafter until 89 y of age. Serum concentrations of homocysteine, vitamin B-12, and folic acid were measured at 85 and 89 y of age. Cross-sectional associations between serum concentrations and cognition were assessed at 85 and 89 y of age. The association between baseline serum concentrations and subsequent longitudinal cognitive decline was assessed with the use of mixed linear models. RESULTS: In the cross sectional analyses, serum concentrations of homocysteine and folic acid were significantly associated with cognitive performance, but serum concentrations of vitamin B-12 were not. In the longitudinal analyses, there were no significant associations of serum concentrations of homocysteine, vitamin B-12, or folic acid with rate of cognitive decline. CONCLUSIONS: Elevated serum concentrations of homocysteine and reduced folic acid are associated with cognitive impairment in elderly persons but do not predict an increased rate of cognitive decline. The association of high serum concentrations of homocysteine and low folic acid with cognitive impairment in old age is likely to be a consequence of disease and not a contributory cause. PMID- 16210719 TI - Weight change and the conservation of lean mass in old age: the Health, Aging and Body Composition Study. AB - BACKGROUND: Weight loss may contribute to the loss of lean mass with age. OBJECTIVE: The objective was to evaluate the relation between weight loss or weight gain and changes in lean mass and fat mass in older adults. DESIGN: We observed changes in weight and body composition during a 4-y period in 2163 men (47%) and women (53%) aged 70-79 y in the Health, Aging and Body Composition Study cohort. Whole-body and appendicular bone-free lean mass and fat mass were measured by using dual-energy X-ray absorptiometry. RESULTS: Weight loss and weight gain were common. In both weight losers and weight gainers, changes in lean mass as a percentage of initial lean mass were substantially smaller than changes in fat mass as a percentage of initial fat mass. However, the difference between the change in lean mass and that in fat mass was more pronounced with weight gain than with weight loss, especially in men. Small amounts of lean loss and fat gain were noted with weight stability. In multivariate models, weight loss was strongly associated with lean mass loss in both men and women, especially in men whose weight loss was concurrent with a hospitalization. CONCLUSIONS: With weight change, a greater proportion of lean mass than of fat mass was conserved, but, especially in older men, significantly more lean mass was lost with weight loss than was gained with weight gain. These findings suggest that weight loss, even with regain, could accelerate sarcopenia in older adults. PMID- 16210720 TI - Plasma carotene and alpha-tocopherol in relation to 10-y all-cause and cause specific mortality in European elderly: the Survey in Europe on Nutrition and the Elderly, a Concerted Action (SENECA). AB - BACKGROUND: Only a few observational studies have related plasma carotene and alpha-tocopherol to mortality in elderly subjects. OBJECTIVE: The objective was to study the association of plasma carotene (alpha-and beta-carotene) and alpha tocopherol with all-cause and cause-specific mortality in elderly subjects who participated in a European prospective study. DESIGN: Plasma concentrations of carotene and alpha-tocopherol were measured in 1168 elderly men and women. After a follow-up period of 10 y, 388 persons had died. The association between plasma antioxidants and mortality was analyzed by using Cox proportional hazard models. To put our results in context, we performed a meta-analysis of 5 studies on plasma antioxidants and all-cause mortality in elderly populations. RESULTS: Plasma carotene concentrations were associated with a lower mortality risk [adjusted rate ratio (RR) for an increment of 0.39 micromol/L: 0.79; 95% CI: 0.70, 0.89]. This lower mortality risk was observed for both cancer (RR: 0.59; 95% CI: 0.44, 0.79) and cardiovascular disease (RR: 0.83; 95% CI: 0.70, 1.00). The lower risk of cardiovascular death was confined to those with a body mass index (in kg/m2) <25 (RR: 0.67; 95% CI: 0.49, 0.94). Plasma concentrations of alpha-tocopherol were not associated with all-cause or cause-specific mortality. The results for both plasma antioxidants and all-cause mortality were confirmed by the meta-analysis. CONCLUSIONS: This prospective study suggests that high plasma concentrations of carotene are associated both with lower mortality from all causes and with cancer in the elderly. For cardiovascular mortality, the inverse association was confined to elderly with body mass indexes <25. PMID- 16210721 TI - Relation between dietary n-3 and n-6 fatty acids and clinically diagnosed dry eye syndrome in women. AB - BACKGROUND: Dry eye syndrome (DES) is a prevalent condition, but information on risk or protective factors is lacking. OBJECTIVE: We aimed to determine the association between the dietary intake and ratio of n-3 and n-6 fatty acids (FAs) and DES occurrence. DESIGN: Of the 39876 female health professionals in the Women's Health Study (WHS), 32470 women aged 45-84 y who provided information on diet and DES were cross-sectionally studied. We assessed FA intakes by using a validated food-frequency questionnaire and assessed DES by using self-reports of clinically diagnosed cases. Of the sample, 1546 (4.7%) subjects reported DES. We used logistic regression models to estimate the odds ratios (ORs) and 95% CIs to describe the relation of FA intake with DES. RESULTS: After adjustment for demographic factors, hormone therapy, and total fat intake, the OR for the highest versus the lowest quintile of n-3 FAs was 0.83 (95% CI: 0.70, 0.98; P for trend = 0.05). A higher ratio of n-6 to n-3 FA consumption was associated with a significantly increased risk of DES (OR: 2.51; 95% CI: 1.13, 5.58) for >15:1 versus <4:1 (P for trend = 0.01). In addition, tuna consumption [1 serving was 113 g (4 oz)] was inversely associated with DES (OR: 0.81; 95% CI: 0.66, 0.99 for 2-4 servings/wk; OR: 0.32; 95% CI: 0.13, 0.79 for 5-6 servings/wk versus < or =1 serving/wk; P for trend = 0.005). CONCLUSIONS: These results suggest that a higher dietary intake of n-3 FAs is associated with a decreased incidence of DES in women. These findings are consistent with anecdotal clinical observations and postulated biological mechanisms. PMID- 16210722 TI - High-fat dairy food and conjugated linoleic acid intakes in relation to colorectal cancer incidence in the Swedish Mammography Cohort. AB - BACKGROUND: High-fat dairy foods contain many potentially anticarcinogenic factors, including conjugated linoleic acid (CLA). However, few epidemiologic studies have specifically evaluated high-fat dairy food consumption, and none have evaluated CLA intake, in relation to colorectal cancer risk. OBJECTIVE: The aim of this study was to prospectively examine the associations of long-term high fat dairy food consumption and CLA intake and the incidence of colorectal cancer. DESIGN: Our study population consisted of 60 708 women aged 40-76 y who participated in the Swedish Mammography Cohort. The women's consumption of high fat dairy foods was assessed at baseline, which was from 1987 to 1990, and again in 1997. RESULTS: We ascertained 798 incident cases of colorectal cancer during an average 14.8 y of follow-up. After adjustment for age and other potential confounders, the women who consumed > or =4 servings of high-fat dairy foods/d (including whole milk, full-fat cultured milk, cheese, cream, sour cream, and butter) had a multivariate rate ratio of colorectal cancer of 0.59 (95% CI: 0.44, 0.79; P for trend = 0.002) when compared with the women who consumed <1 serving/d. Each increment of 2 servings of high-fat dairy foods/d corresponded to a 13% reduction in the risk of colorectal cancer (multivariate rate ratio: 0.87; 95% CI: 0.78, 0.96). For CLA, the multivariate rate ratio of colorectal cancer in a comparison of the 2 extreme quartiles of intake was 0.71 (95% CI: 0.55, 0.91; P for trend = 0.004). CONCLUSION: These prospective data suggest that high intakes of high-fat dairy foods and CLA may reduce the risk of colorectal cancer. PMID- 16210723 TI - Habitual physical activity in children: the role of genes and the environment. AB - BACKGROUND: Understanding the factors that contribute to physical inactivity in children is important because sedentary behavior strongly relates to metabolic disorders such as obesity and diabetes. OBJECTIVE: We aimed to quantify the genetic and environmental influences on physical activity energy expenditure (PAEE) in 100 sex-concordant dizygotic (n = 38) and monozygotic (n = 62) twin pairs aged 4-10 y. DESIGN: Resting metabolic rate (RMR) was assessed by using respiratory gas exchange, total energy expenditure (TEE) by using doubly labeled water, and body composition by using dual-energy X-ray absorptiometry. Structural equation modeling was used to partition the phenotypic variance into additive genetic (a2) and common (c2) and unshared (e2) environmental components. RESULTS: Because PAEE [TEE - (RMR + 0.1 x TEE)] depends on body weight, which is highly heritable, we tested several models: 1) after adjustment for age, sex, ethnicity, study date, season, and weight, a2 explained none of the phenotypic variance in PAEE (95% CI: 0%, 38%), whereas c2 and e2 accounted for 69% (33%, 77%; P = 0.001) and 31% (23%, 39%; P < 0.001) of the variance, respectively; 2) after adjustment for the cofactors in model 1, a2 explained 19% of the phenotypic variance in TEE (0%, 60%; P = 0.13), whereas c2 and e2 accounted for 59% (16%, 79%; P = 0.007) and 23% (17%, 31%; P < 0.0001) of the variance, respectively; 3) in models adjusted as above (excluding weight), a2 explained no variance in physical activity level (TEE/RMR) (0%, 32%; P = 0.50), whereas c2 and e2 explained 65% (34%, 60%; P = 0.001) and 35% (28%, 45%; P < 0.0001) of the variance, respectively. CONCLUSIONS: Our data suggest that the familial resemblance in physical activity in these children is explained predominantly by shared environmental factors and not by genetic variability. PMID- 16210724 TI - The paradox of the "body mass index paradox" in dialysis patients: associations of adiposity with inflammation. PMID- 16210728 TI - One generation of n-3 polyunsaturated fatty acid deprivation increases depression and aggression test scores in rats. AB - Male rat pups at weaning (21 days of age) were subjected to a diet deficient or adequate in n-3 polyunsaturated fatty acids (n-3 PUFAs) for 15 weeks. Performance on tests of locomotor activity, depression, and aggression was measured in that order during the ensuing 3 weeks, after which brain lipid composition was determined. In the n-3 PUFA-deprived rats, compared with n-3 PUFA-adequate rats, docosahexaenoic acid (22:6n-3) in brain phospholipid was reduced by 36% and docosapentaenoic acid (22:5n-6) was elevated by 90%, whereas brain phospholipid concentrations were unchanged. N-3 PUFA-deprived rats had a significantly increased (P = 0.03) score on the Porsolt forced-swim test for depression, and increased blocking time (P = 0.03) and blocking number (P = 0.04) scores (uncorrected for multiple comparisons) on the isolation-induced resident-intruder test for aggression. Large effect sizes (d > 0.8) were found on the depression score and on the blocking time score of the aggression test. Scores on the open field test for locomotor activity did not differ significantly between groups, and had only small to medium effect sizes. This single-generational n-3 PUFA deprived rat model, which demonstrated significant changes in brain lipid composition and in test scores for depression and aggression, may be useful for elucidating the contribution of disturbed brain PUFA metabolism to human depression, aggression, and bipolar disorder. PMID- 16210729 TI - Janus kinase 2 modulates the lipid-removing but not protein-stabilizing interactions of amphipathic helices with ABCA1. AB - ABCA1 mediates the transport of cellular cholesterol and phospholipids to HDL apolipoproteins. Apolipoprotein A-I (apoA-I) interactions with ABCA1-expressing cells elicit several responses, including removing cellular lipids, stabilizing ABCA1 protein, and activating Janus kinase 2 (JAK2). Here, we used synthetic apolipoprotein-mimetic peptides to characterize the relationship between these responses. Peptides containing one amphipathic helix of L- or D-amino acids (2F, D-2F, or 4F) and a peptide containing two helices (37pA) all promoted ABCA1 dependent cholesterol efflux, competed for apoA-I binding to ABCA1-expressing cells, blocked covalent cross-linking of apoA-I to ABCA1, and inhibited ABCA1 degradation. 37pA was cross-linked to ABCA1, confirming the direct binding of amphipathic helices to ABCA1. 2F, 4F, 37pA, and D-37pA all stimulated JAK2 autophosphorylation. Inhibition of JAK2 greatly reduced peptide-mediated cholesterol efflux, peptide binding to ABCA1-expressing cells, and peptide cross linking to ABCA1, indicating that these processes require an active JAK2. In contrast, apoA-I and peptides stabilized ABCA1 protein even in the absence of an active JAK2, implying that this process is independent of JAK2 and lipid efflux promoting binding of amphipathic helices to ABCA1. These findings show that amphipathic helices coordinate the activity of ABCA1 by several distinct mechanisms that are likely to involve different cell surface binding sites. PMID- 16210731 TI - Sexual murderers' implicit theories. AB - Interviews with 28 sexual murderers were subjected to grounded theory analysis. Five implicit theories (ITs) were identified: dangerous world, male sex drive is uncontrollable, entitlement, women as sexual objects, and women as unknowable. These ITs were found to be identical to those identified in the literature as being present in rapists. The presence of dangerous world and male sex drive is uncontrollable were present, or absent, such that three groups could be identified: (a) dangerous world plus male sex drive is uncontrollable; (b) dangerous world, in the absence of male sex drive is uncontrollable; (c) male sex drive is uncontrollable in the absence of dangerous world. These three groups were found to differ in motivation: (a) were motivated by urges to rape and murder; (b) were motivated by grievance, resentment and/or anger toward women; (c) were motivated to sexually offend but were prepared to kill to avoid detection, or secure compliance. PMID- 16210730 TI - Preliminary evidence for an automatic link between sex and power among men who molest children. AB - Understanding critical motivational processes of sexual offenders may ultimately provide important clues to more effective treatments. Implicit, automatic cognitive processes have received minimal attention; however, a lexical decision experiment revealed automatic links between the concepts of power and sex among participants who self-reported attraction to sexual aggression. The current study replicates this experiment with a group of male child molesters and forensic and analogue controls. Subliminally presented sex words elicited a facilitation effect for power words among child molesters only; that is, sex to power associations were evident, as well as a trend for the reverse. These results provide preliminary evidence for an automatic sex-power association in child molesters and may point to a crucial pathological link in the cognitive schemata of sex offenders. As well, the current study suggests that paradigms from cognitive psychology may contribute to multimodal (risk) assessment of sexual offenders. PMID- 16210732 TI - Predictors of trauma symptomatology in sexually abused adolescents: a 6-month follow-up study. AB - This study examines the natural course of trauma-specific symptoms 6 months after disclosure. Furthermore, this study investigates whether severity and type of abuse (intrafamilial or extrafamilial sexual abuse), negative appraisals, coping strategies, and crisis support measured at time of disclosure can be predictive of trauma symptoms 6 months later. Sixty-five sexually abused Flemish adolescents are reassessed 6 months after disclosure. Information from the participants is obtained through self-report questionnaires. Forty-six percent of the adolescents report clinically significant trauma symptoms. Although internalizing symptoms significantly decreases after 6 months, externalizing symptoms persist. Type or severity of the abuse does not account for differences in symptomatology. Two predictors of ongoing trauma symptomatology are identified: postdisclosure trauma symptomatology and a lack of initial crisis support. Information on the victims' postdisclosure symptomatology as well as information on the initial received social support is critical in understanding which abused adolescents are most at risk for poor outcomes in the long term. PMID- 16210733 TI - Child physical abuse: prevalence, characteristics, predictors, and beliefs about parent-child violence in South Asian, Middle Eastern, East Asian, and Latina women in the United States. AB - The present study examined the prevalence, characteristics, beliefs, and demographic predictors of parent-child physical violence among South Asian, Middle Eastern, East Asian, and Latina women in the United States. Two hundred fifty-one college-educated women from a middle to high SES (South Asian/Middle Eastern, n = 93; East Asian,n = 72; Latina,n = 86) completed a self-report survey on childhood experiences and beliefs regarding physical abuse. Seventy-three percent of the South Asian and Middle Eastern sample, 65% of the East Asian sample, and 78% of the Latina sample reported experiencing at least one type of physical abuse. Significant differences in characteristics and perpetrators of abuse were found across groups. Demographic factors did not predict physical abuse. Experiencing physical abuse was the only predictor for acceptance of physical discipline and as a parental privilege or right across groups. Implications of alternate cultural models of family violence based on beliefs and exposure to violence are discussed. PMID- 16210734 TI - Animal cruelty motivations: assessing demographic and situational influences. AB - Few studies have examined childhood and adolescent animal cruelty motives. Using a sample of 261 inmates surveyed at both medium and maximum security prisons ina southern state, the present study examined the impact of demographic attributes and situational factors relating specifically to a range of animal cruelty motivations. Almost half of the inmates who engaged in animal abuse reported committing some of the acts out of anger, whereas more than a third did so for fun. Regression analyses revealed that the most statistically salient variable in 7 of the 10 motivational models was whether animal cruelty was committed alone. Respondents who reported hurting or killing animals alone were more likely to commit the acts out of anger but less likely to have committed them to impress others, for sex, or to imitate others. PMID- 16210735 TI - Bedroom rape: sequences of sexual behavior in stranger assaults. AB - This article examines the sequential, temporal, and interactional aspects of sexual assaults using sequential analysis. Fourteen statements taken from victims of bedroom-based assaults were analyzed to provide a comprehensive account of the behavioral patterns of individuals in sexually charged conflict situations. The cases were found to vary in the sexual severity of assault, distinguishing a variety of motivations and behavioral repertoires of offender and victim. Two quite distinct styles of offense were identified: multiple and single, which may have very different implications for research into rapist taxonomies and rape prevention strategies. PMID- 16210736 TI - Depressive symptoms in women experiencing intimate partner violence. AB - The study was a cross-sectional examination of African American women positive for intimate partner violence (IPV) who presented to the medical or psychiatric emergency department (ED) for treatment. African American women with a recent history of IPV who presented following an attempted suicide (n = 100) were compared to demographically comparable African American women who were IPV positive who had not attempted suicide and presented for treatment of another condition(n= 100). Women completed face-to-face interviews on several measures, including demographics and the Beck Depression Inventory-II (BDI-II). Overall, there were no demographic differences between cases and controls. Attempters reported statistically significant higher scores on all 21 BDI-II items than did nonattempters. Four BDI-II items had effect size values in the medium range: sadness, self-dislike, suicidal thoughts, and feelings of worthlessness. These four items can be used as a brief screen in the ED to detect female patients positive for IPV at increased risk for suicidal behavior. PMID- 16210737 TI - Traumatic stress symptoms in women exposed to community and partner violence. AB - Prior research documents increased trauma symptoms associated with exposure to violence, primarily by examining types of violence separately. This study extends prior research by examining traumatic stress symptoms associated with two types of violence exposure, community violence and partner violence. A sample of 90 low income African American women from an urban area completed measures assessing exposure to community violence, partner violence, and trauma symptoms. Exposure to community violence and partner violence were associated with increased reporting of trauma symptoms. Participants who experienced high levels of exposure to both types of violence reported more trauma symptoms than women who were exposed to only one type of violence or neither type of violence. The results suggest that the accumulation of exposures to violence is linked with greater distress. Thus, interventions with women exposed to violence should assess violence exposure in multiple domains and attend to the implications of multiple exposures to violence. PMID- 16210738 TI - The relationship between life satisfaction, risk-taking behaviors, and youth violence. AB - This study builds on existing criminological theories and examines the role of life satisfaction and self-control in explaining youth violence. Using data from a stratified cluster sample of 5,414 public high school students who responded to the South Carolina Youth Risk Behavior Survey, the study examines the relationship between adolescents'perceptions of life satisfaction, behavioral risky acts, and self-reported acts of violence. Analyses indicate that higher levels of life satisfaction are associated with lower violence. Participation in work and involvement in health-related risk-taking behaviors pertaining to sex, drugs, and alcohol are also associated with increased violence. The implications of these findings for criminological theory and for school-based violence prevention programs are discussed. PMID- 16210739 TI - Challenges for global nursing. PMID- 16210740 TI - Cross-cultural analysis for conceptual understanding: English and Spanish perspectives. AB - Culture and primary language provides the context for understanding between the patient and the nurse and therefore is part of the foundation for nursing care. Knowledge development in nursing is predicated on mutual understanding and interpretation of language. Concept development has been identified as one of the approaches to the development of nursing knowledge. Since the process of concept analysis is based on understanding a phenomenon through language and experiences, this process is culture-bound and language-specific. The purpose of this column is to discuss the value of interviews with people from two different cultures resulting in an attempt to develop nursing knowledge with international relevance. An example of an analysis of a concept, worry, is provided, and the effects of culture and language on this nursing concept are discussed. The analysis was the result of collaboration among nurses from Mexico and the United States of America. PMID- 16210741 TI - The unitary field pattern portrait research method: facets, processes, and findings. PMID- 16210742 TI - The human becoming modes of inquiry: emerging sciencing. PMID- 16210743 TI - The metaphor of nurse as guest with ethical implications for nursing and healthcare. AB - Current healthcare advertising and customer relations terminology acknowledge that healthcare providers, including nurses, are to act as hosts for persons who enter into healthcare agencies and institutions. Indeed, much has been written aligning nursing and other healthcare services with consumer-oriented roles of the hospitality service industry commonly associated with hotels and restaurants. From a human becoming perspective, this article discusses possible ethical, administrative, and practice implications of nurses acting as guests entering into the lives of those we serve. PMID- 16210744 TI - Caring science: belonging before being as ethical cosmology. PMID- 16210745 TI - The influence of the human becoming theory on teaching-learning stories in Hangzhou, China. AB - As a nurse educator, human becoming theory was used to enhance my understanding of my experience with students in Hangzhou, China. I describe my initial tension in living simultaneously the uncertainty and the certainty when I was faced with a strange but yet familiar teaching situation. The tension has however enabled me to cross from a traditional teaching approach to a human becoming teaching learning paradigm where the students were given opportunities to share their practice encounters with patients and families by narrating their lived experiences in a clinical reasoning course. As their meanings of nursing surfaced, the students began to learn about who they were, who they are becoming, and who they want to be as nurses, teachers, and human beings. PMID- 16210746 TI - Nursing as caring theory: living caring in practice. AB - This column features a report of a research project in a community hospital. Staff members were given opportunities to engage and act on the ideas of Boykin and Schoenhofer in their nursing as caring theory. Feedback from patients, family members, and all staff who touched patients informed the creation of a new model of care that positively transformed the workplace in the hospital setting. Strategies that were implemented to guide the values of nursing as caring theory are described. The result of the project was an environment that nurtures persons in their caring and growing in caring while illuminating the fullness of nursing. PMID- 16210748 TI - Persevering through a difficult time during the SARS outbreak in Toronto. AB - The purpose of this study was to describe the experience of persevering through a difficult time for patients, family members of patients, nurses, and allied health professionals during the severe acute respiratory syndrome outbreak. van Kaam's phenomenological research method, with the human becoming theory as the theoretical perspective, was used to gather and analyse data from 63 participants who agreed to describe a situation that illuminated their experience of persevering through a difficult time (either online or using a voicemail system). Data gathering occurred in early April 2003 in the midst of the severe acute respiratory syndrome outbreak in Toronto, Canada. The finding was the structural definition, persevering through a difficult time is dispiriting trepidation arising with witnessing suffering. It is a smothering connectedness with sequestering protection as unsettling contentment emerges amid unburdening hope. It sheds light on what is important for preparing for possible future outbreaks of this and other infectious diseases. PMID- 16210749 TI - Health as expanding consciousness: seven women living with multiple sclerosis or rheumatoid arthritis. AB - The meaning of health as expanding consciousness is explored through stories of seven women who developed multiple sclerosis or rheumatoid arthritis during their lives. Using Newman's hermeneutic-dialectic approach, unstructured interviews were conducted over a 2-year period. Analysis and interpretation of narratives concerning person-environment interactions revealed turning points and separate choice points before four new ways of living including finding simple pleasures, being positive, gaining self-control, and self-differentiation, were found. Support for Newman's stages of expanding consciousness and more comprehensive descriptions of self-transcendence in space and time are presented. Implications for theory development and theory-guided practice are offered. PMID- 16210750 TI - Exploring underlying life patterns of women with multiple sclerosis or rheumatoid arthritis: comparison with NANDA dimensions. AB - In Newman's theory, disease is one of many manifestations of underlying pattern and its existence provides meaningful information about person-environment interactions. Underlying patterns manifest differently over time, so clues to their understanding can be found within life stories. Further interpretation subsequent to illustrating expanding consciousness for seven women living with multiple sclerosis or rheumatoid arthritis suggested six underlying patterns expressed in theoretical terms as energy-fatigue, giving-receiving, rejecting accepting, vulnerability-resilience, control-release, and being silent-speaking out. Discussion and comparison with the North American Nursing Diagnosis Association's dimensions for assessment of human response patterns illustrates how nurses caring for women could identify and use underlying patterns in practice. PMID- 16210751 TI - Considering and enjoying tomorrow: global aging through a human becoming lens. AB - The aging of the world's population is a global issue of considerable importance to everyone, particularly nursing. Traditional developmental theories and the medical model have either neglected later life or highlighted the losses and deficits associated with advanced age, while more recent research on aging and the human becoming theory has provided a more balanced view. In this column, information from health and social sciences, the human becoming school of thought, and international political debates about aging are interwoven. Some attention is also given to the issues of the retirement and aging of the nurses. PMID- 16210752 TI - Vulnerable research participants: anyone may qualify. AB - Although the Code of Federal Regulations has provided guidelines regarding research involving vulnerable participants, persons not traditionally identified may also be considered vulnerable at a given time due to emergent or acute illness, end-of-life needs, or circumstances encountered that challenge capacity to provide informed consent. In this column, the author presents an overview of institutional review board roles, responsibilities, processes, and suggestions regarding assessment of vulnerability and capacity to consent that is responsive to current societal and healthcare needs. PMID- 16210753 TI - K-site splitting in KTiOPO4 at room temperature. AB - The room-temperature structure of potassium titanyl phosphate (KTiOPO4, KTP) with Pna2(1) symmetry has been studied by means of synchrotron radiation. Each of the two crystallographically unique K1 and K2 cations is split over two sites that are shifted along the c direction by 0.287 (13) and 0.255 (13) A for the K1a/b and K2a/b pairs, respectively. The refined populations of the minor K1b and K2b sites are 0.102 (12) and 0.132 (17), respectively. It is shown that accurate high resolution synchrotron data (Rmerged = 0.015 for 25 010 reflections, 9456 unique, sintheta/lambda limit > 1.0) are required for the determination of a reliable structure model. PMID- 16210754 TI - Ag2Hg2(TeO4)3. AB - Red single crystals of disilver(I) dimercury(II) tris[tetraoxotellurate(VI)], Ag2Hg2(TeO4)3, were obtained under hydrothermal conditions at 523 K. The structure is built up of infinity1[(TeO2/1O4/2)({TeO2/1O2/2}2O4/2)] chains, with an overall composition [TeO4]2-, that run parallel to the crystallographic a axis. Distorted AgO6 and HgO6 polyhedra (the latter with two short and nearly collinear Hg-O bonds) link the tellurate chains into a three-dimensional network. Except for one Te atom situated on an inversion center, all atoms occupy general positions. PMID- 16210755 TI - Ca2CuTe4O10Cl2, a new synthetic tellurium(IV) oxochloride. AB - Single crystals of dicalcium copper tetratellurium decaoxide dichloride, Ca2CuTe4O10Cl2, were synthesized via a transport reaction in sealed evacuated quartz glass tubes. The building units of the structure are irregular CaO7 polyhedra, centrosymmetric CuO4Cl2 octahedra and two crystallographically distinct TeO4E distorted bipyramids (E being the 5s2 lone pair of TeIV). The TeO4E and CuO4Cl2 polyhedra together form planes that are connected by the Ca atoms. The CuO4Cl2 octahedra are isolated from each other by the other building units. PMID- 16210756 TI - [Pd8(CH3COO)8(NO)8]: solution from X-ray powder diffraction data. AB - The water-insoluble title compound, octakis(mu-acetato-kappa2O:O)octakis(mu nitroso-kappa2N:O)octapalladium(II), [Pd8(CH3COO)8(NO)8], was precipitated as a yellow powder from a solution of palladium nitrate in nitric acid by adding acetic acid. Ab initio crystal structure determination was carried out using X ray powder diffraction techniques. Patterson and Fourier syntheses were used for atom locations, and the Rietveld technique was used for the final structure refinement. The crystal structure is of a molecular type. The skeleton of the [Pd8(CH3COO)8(NO)8] molecule is constructed as a tetragonal prism with Pd atoms at the vertices. The eight NO- groups are in bridging positions along the horizontal edges of the prism. The N and O atoms of each nitroso group coordinate two different Pd atoms. The vertical edges present PdPd contacts with a short distance of 2.865 (1) A. These Pd atoms are bridged by a pair of acetate groups in a cis orientation with respect to each other. The complex has crystallographically imposed 4/m symmetry; all C atoms of the acetate groups are on mirror planes. The unique Pd atom lies in a general position and has square planar coordination, consisting of three O and one N atom. PMID- 16210757 TI - Triaquabenzoatocalcium(II) monobenzoate: coordination polymer chains linked into a two-dimensional framework by hydrogen bonds. AB - In the title complex, catena-poly[[[diaquacalcium(I)]-mu2-aqua-mu3-benzoato kappa4O:O,O':O'] benzoate], {[Ca(C7H5O2)(H2O)3](C7H5O2)}n, obtained by the reaction of CaCl2 and potassium benzoate in water, the Ca atom is eight coordinated by four carboxylate O atoms and four water molecules. The structure consists of polymeric {[Ca(C6H5COO)(H2O)3]+}infinity chains alternating with layers of uncoordinated C6H5COO- anions. The nearly planar anions are linked to the chain by short hydrogen bonds to form a two-dimensional network. PMID- 16210758 TI - Co-crystallized cis and trans isomers of dichlorobis(2-picolylamine)iron(II). AB - The octahedral cis and trans isomers of dichlorobis(2-picolylamine)iron(II), [FeCl2(C6H8N2)2], co-crystallize in a 1:1 ratio. The cis isomer lies on a twofold axis, whereas the trans isomer lies on an inversion centre. The structure is fully ordered, with both Fe atoms in a pure high-spin state. The Fe, Cl and N(H2) atoms of both isomers lie in the same plane, allowing all Cl and amine H atoms to be engaged in extensive two-dimensional hydrogen bonding. The hydrogen-bonded layers are interconnected through pi-pi interactions between the pyridine rings. Searches in the Cambridge Structural Database uncover very few examples of such isomer co-existence. PMID- 16210759 TI - Poly[[cobalt(II)-di-mu3-3,5-diaminobenzoato-kappa3N:N':O] monohydrate]. AB - In the title compound, {[Co(C7H7N2O2)2].H2O}n, the CoII atom lies on an inversion centre and has octahedral geometry, defined by two O atoms in axial positions and four N atoms in equatorial sites from six different 3,5-diaminobenzoate ligands. Each 3,5-diaminobenzoate anion acts as a mu3-bridging ligand, linking three adjacent CoII ions through one O atom and two N atoms to form a three-dimensional coordination polymer. PMID- 16210760 TI - catena-Poly[[[4-bromo-2-(2-pyridylmethyliminomethyl)phenolato]zinc(II)]-mu chloro]. AB - The title complex, [Zn(C13H10BrN2O)Cl]n, is a chloride-bridged polynuclear zinc(II) compound. Each ZnII ion is five-coordinated in a square-pyramidal configuration, with one O and two N atoms of one Schiff base and one bridging Cl atom defining the basal plane, and another bridging Cl atom occupying the apical position. The novelty of the compound lies in the bridging by chlorine of two square-pyramidal Zn atoms, so that the bridging atom is apical for one Zn ion and basal for the other. This structural arrangement has not been observed before. The linked moieties form polymeric zigzag chains running along the a axis. PMID- 16210761 TI - Polymorphism in an unexpected caesium complex of 5-hydroxyhydurilic acid. AB - Vigorous reaction of barbituric acid with caesium hydroxide in water resulted in an unexpected coupling product, 5-hydroxyhydurilic acid, complexed to caesium, giving poly[[caesium-mu5-5-hydroxyhydurilato] hemihydrate], {[Cs(C8H5N4O7)].0.5H2O}n. This was obtained in two different polymorphic forms, depending on the method of crystal growth. Slow solvent evaporation yielded an orthorhombic polymorph, (I), which crystallized in the space group Pnab (non standard setting of Pbcn), with the uncoordinated water molecule lying on a crystallographic twofold axis. The water molecule is sandwiched into cavities within the structure and is securely held in place by N-HO and O-HO hydrogen bonding. Polymorph (II) is monoclinic, although the unit-cell parameters are similar to those of polymorph (I), and it crystallizes in the space group C2/c, with the uncoordinated water molecule again lying on the twofold axis and secured by hydrogen bonding. The differences between the two polymorphs, both of which have nine-coordinate caesium and a similar first-shell environment of all structural components, are in the overall arrangement of the cations, anions and water molecules, and in the shape of the cavities, revealed by inspecting and comparing crystal-packing diagrams. PMID- 16210762 TI - Cytosine-containing hybrid dipeptides: N-[2-(4-amino-2-oxo-1,2-dihydropyrimidin-1 yl)propionyl]-L-phenylalanine N-[2-(4-amino-2-oxo-1,2-dihydropyrimidin-1 yl)propionyl]-L-serine monohydrate and N-[2-(4-amino-2-oxo-1,2-dihydropyrimidin-1 yl)propionyl]-L-lysine. AB - The title compounds are cytosine-incorporating hybrid dipeptides showing affinities for 9-ethyl-7-methylguanine (7mG). Four molecules of the L phenylalanine (L-Phe) derivative, C16H14N4O4, are present in the asymmetric unit, with similar folded conformations but with slightly different torsion angles involving the L-Phe group. The L-serine (L-Ser) derivative crystallizes as a monohydrate, C10H14N4O5.H2O, the two independent molecules having extended conformations, whereas the two independent molecules of the L-lysine (L-Lys) system in the final compound, C13H21N5O4, are folded. The cytosine-cytosine base pair (pyrimidine NN interactions) was observed only for the L-lysine derivative. Conformational comparisons with previous structures of cytosine hybrid dipeptides may show the relationships between side-chain position and binding phases for 7mG. PMID- 16210763 TI - 4,5-Bis(furoylsulfanyl)-1,3-dithiole-2-thione. AB - The title compound, C13H6O4S5, possesses crystallographically imposed mirror symmetry, with the atoms of the C=S group lying on the mirror plane. It is an example of the general formula [RCO]2(dmit), where R is a furan ring and dmit is 2-thioxo-1,3-dithiole-4,5-dithiolate. The components exhibit some polarization of their molecular-electronic structure. The dmit and furan moieties exhibit a high degree of conjugation, as the introduction of C=O connecting the conjugated furan (donor) and dmit (acceptor) rings forms a good conjugated system with high delocalization. A polar three-dimensional framework is built from a combination of intermolecular contacts, namely SS interactions and C-HO hydrogen bonding. The structural characteristics lead to good second-order non-linear optical properties. PMID- 16210764 TI - Hydrogen-bonding patterns in pyrimethaminium dinitrate. AB - The title compound, 2,4-diamino-5-(4-chlorophenyl)-6-ethylpyrimidine-1,3-diium dinitrate, C12H15ClN42+.2NO3-, contains two crystallographically independent pyrimethamine (PMN) molecules, which differ in the relative orientations of the pyrimidine and benzene rings and of the ethyl substitutents. In both pyrimethamine molecules, all the pyrimidine N atoms are protonated, unlike most related compounds, in which only one pyrimidine N atom is protonated. The two pyrimethamine moieties are bridged by a variety of N-HO(nitrate) interactions, including some three-centre hydrogen bonds. PMID- 16210765 TI - Isatin 3-semicarbazone and 1-methylisatin 3-semicarbazone. AB - The two title semicarbazones, namely 2,3-dihydro-1H-indole-2,3-dione 3 semicarbazone, C9H8N4O2, (I), and 1-methyl-2,3-dihydro-1H-indole-2,3-dione 3 semicarbazone, C10H10N4O2, (II), show the same configuration, viz. Z around the imine C=N bond and E around the C(O)-NH2 bond, stabilized by two intramolecular hydrogen bonds. The presence of a methyl group on the isatin N atom determines the difference in the packing; in (I), the molecules are linked into chains which lie in the crystallographic (102) plane and run perpendicular to the b axis, while in (II), the molecules are arranged to form helices running parallel to a crystallographic screw axis in the a direction. PMID- 16210766 TI - Hydrogen bonding and pi-pi interactions in a laminar structure: benzene-1,3 dicarboxylic acid-4-methylpyridine (1/2). AB - The title complex, C8H6O4.2C6H7N, consists of two crystallographically independent 1:2 clusters of benzene-1,3-dicarboxylic acid and 4-methylpyridine. Each cluster, the components of which are linked by O-H...N hydrogen bonds, is almost planar by alignment of C-H...O hydrogen bonds. Herring-bone ribbons of clusters are formed by other C-H...O hydrogen bonds, and these ribbons are further packed to form a laminar structure by pi-pi interactions. PMID- 16210767 TI - Dimethyl selenoxide. AB - The title molecule, C2H6OSe, has a trigonal-pyramidal structure analogous to that of its sulfur analog, dimethyl sulfoxide (DMSO). The Se-O distance in dimethyl selenoxide (DMSeO) is 1.6756 (16) A [versus S-O of 1.531 (5) A in DMSO], consistent with a highly polar bond. In the solid state, the molecules of DMSeO are linked into centrosymmetric dimers formed by two C-H...O hydrogen bonds. These dimers further aggregate into a ladder-like supramolecular network via two additional intermolecular C-H...O interactions. As a result, each O atom of DMSeO acts as an acceptor of three hydrogen bonds. PMID- 16210768 TI - 1-(2-Methyl-5-phenyl-3-thienyl)-2-(3-methyl-5-phenyl-2-thienyl)-3,3,4,4,5,5 hexafluorocyclopent-1-ene: a novel photochromic hybrid diarylethene. AB - The title compound, C27H18F6S2, a novel photochromic hybrid diarylethene derivative containing 2- and 3-thienyl substituents, is one of the most promising photochromic candidates with shorter wavelength for optical storage and other optoelectronic devices. In the crystal structure, the molecule adopts a photoactive antiparallel conformation. The distance between the two reactive C atoms, i.e. the ring C atoms to which the methyl groups are attached, is 3.430 (4) A. The dihedral angles between the thienyl and adjacent phenyl rings are 26.8 (2) and 33.98 (9) degrees. PMID- 16210769 TI - Three isomeric forms of hydroxyphenyl-2-oxazoline: 2-(2-hydroxyphenyl)-2 oxazoline, 2-(3-hydroxyphenyl)-2-oxazoline and 2-(4-hydroxyphenyl)-2-oxazoline. AB - Crystal structures are reported for three isomeric compounds, namely 2-(2 hydroxyphenyl)-2-oxazoline, (I), 2-(3-hydroxyphenyl)-2-oxazoline, (II), and 2-(4 hydroxyphenyl)-2-oxazoline, (III), all C9H9NO2 [systematic names: 2-(4,5-dihydro 1,3-oxazol-2-yl)phenol, (I), 3-(4,5-dihydro-1,3-oxazol-2-yl)phenol, (II), and 4 (4,5-dihydro-1,3-oxazol-2-yl)phenol, (III)]. In these compounds, the deviation from coplanarity of the oxazoline and benzene rings is dependent on the position of the hydroxy group on the benzene ring. The coplanar arrangement in (I) is stabilized by a strong intramolecular O-H...N hydrogen bond. Surprisingly, the 2 oxazoline ring in molecule B of (II) adopts a 3T4 (C2TC3) conformation, while the 2-oxazoline ring in molecule A, as well as that in (I) and (III), is nearly planar, as expected. Tetramers of molecules of (II) are formed and they are bound together via weak C-H...N hydrogen bonds. In (III), strong intermolecular O-H...N hydrogen bonds and weak intramolecular C-H...O hydrogen bonds lead to the formation of an infinite chain of molecules perpendicular to the b direction. This paper also reports a theoretical investigation of hydrogen bonds, based on density functional theory (DFT) employing periodic boundary conditions. PMID- 16210770 TI - Polymorphic di-2-pyridyl ketone 4-nitrophenylhydrazone (dpknph): the structure of beta-dpknph. AB - A new polymorph of di-2-pyridyl ketone 4-nitrophenylhydrazone [alternative name: N-(dipyridin-2-ylmethylene)-N'-(4-nitrophenyl)hydrazine], C17H13N5O2, isolated from the filtrate of a sonicated acetonitrile solution of dpknph and CdCl2, was found to crystallize in the monoclinic space group P21/c, in contrast to the known form which crystallizes in P21/n. The non-coplanar molecules pack in parallel stacks without any intermolecular hydrogen-bonding interactions. This packing pattern contrasts with the interlocked interdigitated packing seen in the previously known polymorph. PMID- 16210771 TI - An unusual acid: (+/-)-3-benzoyl-1,2-dimethyl-8a-phenyl-2-benzothieno[2,3 b]pyrrole-1,2-dicarboxylic anhydride. AB - The title compound, C27H21NO4S, is a 2-benzothieno[2,3-b]pyrrole derivative with several substituents, present in the crystal as a racemate. The tetracyclic fused ring system shows a 'U-shaped' molecular architecture, since the two rings flanking the central pyrrolidine ring both point in the same direction. PMID- 16210772 TI - Three isomeric (E)-nitrobenzaldehyde nitrophenylhydrazones: chains of rings in isomorphous (E)-2-nitrobenzaldehyde 3-nitrophenylhydrazone and (E)-3 nitrobenzaldehyde 2-nitrophenylhydrazone, and centrosymmetric dimers in (E)-4 nitrobenzaldehyde 2-nitrophenylhydrazone. AB - The isomeric compounds (E)-2-nitrobenzaldehyde 3-nitrophenylhydrazone and (E)-3 nitrobenzaldehyde 2-nitrophenylhydrazone, both C13H10N4O4, are isomorphous and effectively isostructural, and in both, the molecules are disordered across centres of inversion in the space group P21/c. The molecules are linked into complex chains of rings by N-H...O and C-H...O hydrogen bonds. In the isomeric compound (E)-4-nitrobenzaldehyde 2-nitrophenylhydrazone, the fully ordered molecules are linked by N-HO hydrogen bonds into centrosymmetric dimers. PMID- 16210773 TI - pi-Stacked hydrogen-bonded chains of rings in 2,4-difluorobenzaldehyde isonicotinoylhydrazone and hydrogen-bonded sheets in 2,3-dichlorobenzaldehyde isonicotinoylhydrazone. AB - The difluorinated ring in 2,4-difluorobenzaldehyde isonicotinoylhydrazone, C13H9F2N3O, (I), is disordered over two sets of sites with unequal occupancy. The molecules of (I) are linked by a combination of N-H...O and C-H...O hydrogen bonds into chains of rings, which are linked into sheets by a single pi-pi stacking interaction. In 2,3-dichlorobenzaldehyde isonicotinoylhydrazone, C13H9Cl2N3O, (II), the molecules are linked by a combination of N-H...N, C-H...N and C-H...O hydrogen bonds into sheets of R(4)(4)(14) and R(4)(4)(26) rings. PMID- 16210774 TI - Dehydroepiandrosterone sulfate reduces acoustic injury of the guinea-pig cochlea. AB - The present study was performed to determine effects of dehydroepiandrosterone sulfate (DHEAS), a neurosteroid, on acoustic injury. Albino guinea pigs were exposed to a 2 kHz pure tone of 120 or 125 dB sound pressure level for 10 min immediately after intravenous administration of DHEAS. Statistically significant improvement in the compound action potential threshold shifts and in amplitude reduction of distortion-product otoacoustic emissions was observed 1 week after the acoustic overexposure in the animals treated with DHEAS. The present results suggest that DHEAS has a protective effect against acoustic injury of the cochlea. PMID- 16210775 TI - Underlying mechanism of combined effect of methamphetamine and morphine on lethality in mice and therapeutic potential of cooling. AB - An increase in polydrug abuse is a major problem worldwide. A previous study showed that coadministration of methamphetamine and morphine induced lethality in rodents and humans. However, the underlying mechanisms by which the lethality is increased by the coadministration of methamphetamine and morphine have not been fully understood. Therefore, the present study was designed to determine the mechanism of increased lethality induced by methamphetamine and morphine. Coadministered methamphetamine and morphine increased the lethality by more than 70% in BALB/c mice. Pretreatment with NMDA-receptor antagonists, such as MK-801 and 3-((R)-2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (CPP), and benzamide [poly(ADP-ribose) polymerase (PARP) inhibitor] significantly attenuated the increased lethality induced by methamphetamine and morphine. Furthermore, the lethal effect induced by methamphetamine and morphine was completely attenuated by immediate cooling after the coadministration of methamphetamine and morphine. It has been reported that methamphetamine-induced neurotoxicity can be blocked by lowering the temperature, and this effect might be mediated by a reduction of release of free radicals. These results suggest that activation of NMDA receptors and PARP play an important role in the increased lethality induced by methamphetamine and morphine. PMID- 16210776 TI - Sphingosylphosphorylcholine antagonizes proton-sensing ovarian cancer G-protein coupled receptor 1 (OGR1)-mediated inositol phosphate production and cAMP accumulation. AB - Ovarian cancer G-protein-coupled receptor 1 (OGR1), previously proposed as a receptor for sphingosylphosphorylcholine (SPC), has recently been identified as a proton-sensing or extracellular pH-responsive G-protein-coupled receptor stimulating inositol phosphate production, reflecting the activation of phospholipase C. In the present study, we found that acidic pH stimulated cAMP accumulation, reflecting the activation of adenylyl cyclase, in addition to inositol phosphate production in OGR1-expressing cells. The cAMP response was hardly affected by the inhibition of phospholipase C. SPC inhibited the acidification-induced actions in a pH-dependent manner, while no OGR1-dependent agonistic action of SPC was observed. Thus, the dose-response curves of the proton-induced actions were shifted to the right in the presence of SPC regardless of stereoisoform. The antagonistic property was also observed for psychosine and glucosylsphingosine. In conclusion, OGR1 stimulation may lead to the activation of adenylyl cyclase in addition to phospholipase C in response to extracellular acidification but not to SPC. However, SPC and related lysolipids antagonize the proton-induced and OGR1-mediated actions. PMID- 16210777 TI - Perospirone, a novel antipsychotic drug, inhibits marble-burying behavior via 5 HT1A receptor in mice: implications for obsessive-compulsive disorder. AB - Perospirone is a novel atypical antipsychotic drug with dopamine (DA) D(2)- and serotonin (5-hydroxytryptamine, 5-HT) 5-HT(2A)-receptor antagonist, and 5-HT(1A) receptor agonist properties. In the present study, we examined the effect of perospirone on marble-burying behavior, which has been considered an animal model of obsessive-compulsive disorder (OCD), compared with the effects of other antipsychotics such as haloperidol and risperidone. Perospirone at a dose of 10 mg/kg (p.o.) inhibited marble-burying behavior without affecting the locomotor activity in mice. On the other hand, haloperidol (0.1 mg/kg, i.p.) and risperidone (1 mg/kg, p.o.) showed significant suppression of locomotor activity at the dose that inhibited marble-burying behavior. Furthermore, the inhibition of marble-burying behavior by perospirone was antagonized by WAY100135 (10 mg/kg, i.p.), a selective 5-HT(1A)-receptor antagonist. WAY100135 at the same dose also antagonized the inhibition of marble-burying behavior by 8-OH-DPAT (3 mg/kg, i.p.), a selective 5-HT(1A)-receptor agonist. These findings suggest that perospirone may exhibit anti-OCD activity in clinical use and that 5-HT(1A) receptor agonistic activity may be involved in the inhibition of marble-burying behavior by perospirone. PMID- 16210778 TI - New twist on neuronal insulin receptor signaling in health, disease, and therapeutics. AB - Long after the pioneering studies documenting the existence of insulin (year 1967) and insulin receptor (year 1978) in brain, the last decade has witnessed extraordinary progress in the understanding of brain region-specific multiple roles of insulin receptor signalings in health and disease. In the hypothalamus, insulin regulates food intake, body weight, peripheral fat deposition, hepatic gluconeogenesis, reproductive endocrine axis, and compensatory secretion of counter-regulatory hormones to hypoglycemia. In the hippocampus, insulin promotes learning and memory, independent of the glucoregulatory effect of insulin. Defective insulin receptor signalings are associated with the dementia in normal aging and patients with age-related neurodegenerative diseases (e.g., Alzheimer's disease); the cognitive impairment can be reversed with systemic administration of insulin in the euglycemic condition. Intranasal administration of insulin enhances memory and mood and decreases body weight in healthy humans, without causing hypoglycemia. In the hypothalamus, insulin-induced activation of the phosphoinositide 3-kinase pathway followed by opening of ATP-sensitive K+ channel has been shown to be related to multiple effects of insulin. However, the precise molecular mechanisms of insulin's pleiotropic effects still remain obscure. More importantly, much remains unknown about the quality control mechanisms ensuring correct conformational maturation of the insulin receptor, and the cellular mechanisms regulating density of cell surface functional insulin receptors. PMID- 16210779 TI - Transition of the pregnancy rate of bisected bovine embryos after co-transfer with trophoblastic vesicles prepared from in vivo-cultured in vitro-fertilized embryos. AB - Bisected bovine embryos were co-transferred with trophoblastic vesicles (TVs). These TVs were prepared by dissection of conceptuses that were collected by uterine flushing after culture for seven days in the uterus following transfer of embryos derived by in vitro fertilization (IVF). Pregnancy diagnoses were performed twice, between Day 26 and Day 43 and between Day 38 and Day 73 post estrus by ultrasonography. The pregnancy rate was significantly increased at first pregnancy diagnosis when demi-embryos were transferred with TVs (66.7%, 16/24) compared with the control group (34.5%, 10/29) (P < 0.05). Three losses occurred in the co-transfer group between the first and second pregnancy diagnosis. The final pregnancy rates according to delivered offspring were 41.7% (10/24) and 27.6% (8/29), respectively. There were no statistically significant differences between the pregnant and non-pregnant groups with regard to the average diameter of the TVs measured before transfer at three points during the gestation period. The birth weight and gestation lengths of the offspring were almost the same for the co-transfer and control groups. In the co-transfer group, the genetic identities of calves from the separated embryos were not affected by the TVs, as confirmed by parental blood type testing. Delivered offspring in co transferred groups showed normal morphology. In conclusion, the present study indicates that co-transfer of TVs prepared from conceptuses cultured in vivo following transfer of IVF embryos enhances the fertility of demi-embryos during the early stages of pregnancy, as has similarly been shown in previous research for those prepared from in vivo embryos. PMID- 16210780 TI - Immune responses during the peripartum period in dairy cows with postpartum endometritis. AB - Determining the immune responses to the development of endometritis during the peripartum period may assist in the development of more efficient reproductive management regimens for dairy herds. In this study, we compared the peripartum immune responses of dairy cows that develop endometritis by 4 weeks postpartum (n = 11) to cows that did not develop this disease (n = 19). Blood samples were collected 1 week before calving, just after or during calving, and then at weeks 1, 2, 3, and 4 postpartum. Cows that developed endometriris had significantly higher total leukocyte, neutrophil, lymphocyte, and monocyte counts than the control cows (P < 0.05) at all time points. The leukocytes from cows that developed endometritis were significantly less phagocytic than those from control cows at all sampling time points (P < 0.01). The serum TNFalpha concentrations of the control cows decreased linearly from the prepartum time point (P = 0.0029), but the endometritis cows showed a different profile (P > 0.05). As a result, the serum TNFalpha concentrations were greater in the endometritis group (P < 0.01) than in the control group during the third and fourth weeks postpartum. The greater total leukocyte numbers and neutrophil, lymphocyte and monocyte counts, and the maintenance of elevated serum TNFalpha levels in the cows with endometritis may be due to infection in the postpartum period. Furthermore, the decreased phagocytic capacity of leukocytes during the peripartum period, including at the prepartum time point, makes cows more susceptible to postpartum endometritis. PMID- 16210781 TI - Sperm motility, plasma membrane integrity, and binding capacity to homologous zona pellucida of cryopreserved epididymal spermatozoa in the domestic cat. AB - We examined motility, plasma membrane integrity, and binding capacity to homologous zona pellucidae (ZP) of frozen/thawed epididymal cat sperm as a model species for endangered felines. Epididymal spermatozoa from 20 domestic cats were frozen with freezing egg-yolk extender containing 3.0% glycerol in 0.25-ml straws. Post-thaw motility and plasma membrane integrity of the frozen/thawed spermatozoa were 31.8 +/- 2.4% and 32.2 +/- 4.2%, respectively. The frozen/thawed spermatozoa were co-cultured with frozen/thawed immature homologous oocytes with intact ZP for 3 h to examine their ability to bind to the ZP. Sixteen of the 20 frozen/thawed sperm samples demonstrated the ability to bind to ZP. These results indicated that the freezing system for epididymal sperm used in the present study gives appropriate information for banking the genetic resources of wild felid species. PMID- 16210783 TI - Histological study of granulated metrial gland (GMG) cells in beige (DA-bg/bg) rats. AB - To clarify the roles of granulated metrial gland (GMG) cells for successful pregnancy in rats, GMG cells in beige rats (genotype: DA-bg/bg), whose NK cells show lysosomal dysfunction because of abnormalities in cytoplasmic granules, were examined in mid- and late-pregnancy by light and electron microscopies. The GMG cells of beige rats were significantly less in number than those of the two controls (genotypes: DA-bg/+ and DA-+/+) in mid- and late-pregnancy, and this accompanied a low reproductive performance in the beige rats. The size of intracellular granules in the GMG cells of the beige rats was larger than for the two controls on each corresponding day of pregnancy. These results suggest that the activity of rat GMG cells and peripheral NK cells might be influenced by the beige gene, which is involved in reproductive performance. PMID- 16210782 TI - Expression patterns of the implantation-associated genes in the uterus during the estrous cycle in mice. AB - The mRNA expression patterns of EGF, HB-EGF, Amphiregulin, EGF receptor, IGF-1, CSF-1, IL-1 alpha, IL-1 beta, IL-1 receptor type 1, IL-1 receptor antagonist, LIF, COX-1, COX-2, Mucin-1, calcitonin, and rat USAG-1 mouse homologue, all of which are involved in the process of conceptus implantation to the endometrium, were examined during the estrous cycle by means of real-time quantitative PCR. COX-2, HB-EGF, LIF, Mucin-1, CSF-1, IL-1 alpha, IL-1 beta, and IL-1 receptor antagonist were temporally regulated during the estrous cycle and highly expressed during the estrous stage. In the case of COX-1, EGF, IGF-1, and EGF receptor, the highest mRNA expression was during the diestrous stage. In contrast, the rat USAG-1 mouse homologue mRNA expression did not change during the estrous cycle. These results indicate that rat USAG-1 mouse homologue expression at implantation might be specifically regulated by embryonic factors rather than the maternal environment. PMID- 16210784 TI - Effect of cycloheximide on in vitro development of electrically activated feline oocytes. AB - This study was conducted to improve parthenogenetic development in vitro of feline oocytes following a combined activation treatment of electrical stimulation and cycloheximide. In vitro matured (IVM) oocytes were stimulated electrically by a DC electrical pulse of 2 kV/cm for 50 micros. The stimulated oocytes were then incubated in MK-1 medium with or without cycloheximide and subsequently cultured in vitro for 6 days. No significant differences were observed between the two groups with respect to the proportions of cleavage, development to the morula stage, and the cell number of blastocysts. However, exposure of electrically stimulated oocytes to cycloheximide significantly increased the rate of development of the stimulated oocytes into the blastocyst stage compared with oocytes stimulated by electrical stimulation alone (31.0% vs 6.7%). The results from the present study suggested that a single electrical stimulation was insufficient to activate the IVM cat oocytes at 24 h of maturation and that exposure to cycloheximide following electrical stimulation improved the efficacy of the parthenogenetic development of domestic cat oocytes. PMID- 16210785 TI - Radiation-induced apoptosis is independent of caspase-8 but dependent on cytochrome c and the caspase-9 cascade in human leukemia HL60 cells. AB - We investigated the role of the caspase activation cascade in apoptosis induced by ionizing radiation or hydrogen peroxide (H(2)O(2)) in human leukemia HL60 cells. Electron paramagnetic resonance (EPR) spectra revealed that hydroxyl and hydrogen radicals were generated in the culture medium after exposure to radiation or H(2)O(2). Initial accumulation of DNA fragments at 2 h after exposure was delayed in irradiated cells compared with H(2)O(2)-treated cells, although formation of abasic sites immediately after exposure was significantly higher in irradiated cells and similar quantities of hydroxyl radicals were produced under both conditions. Activity assay of caspases revealed that caspase 3, -8 and -9 were activated 2 h after exposure to H(2)O(2), whereas in irradiated cells caspase-3 and -9 activation occurred 4 h after exposure but increased caspase-8 activation was not observed. Release of cytochrome c into cytosol was seen at 2 h after radiation and H(2)O(2) treatment. Radiation did not affect proapoptotic proteins (Bax and Bid), whereas H (2)O(2) increased accumulation of Bax in the mitochondrial membrane 2 h to 6 h after treatment, independently of the truncation of Bid by activated caspase-8. Moreover, treatment with the caspase-8 inhibitor Z-IETD-FMK increased cell survival and prevented accumulation of DNA fragments in H(2)O(2)-treated cells, but not in irradiated cells. These results suggest that, unlike the caspase cascade of H(2)O(2)-induced apoptosis, cytochrome c and caspase-9 are important for the intrinsic pathway of radiation induced apoptosis, independent of caspase-8. PMID- 16210786 TI - Involvement of peroxiredoxin I in protecting cells from radiation-induced death. AB - Peroxiredoxin I (Prx-I), a key member of the peroxiredoxin family, reduces peroxides and equivalents through the thioredoxin system. Our previous work has shown that expression of Prx-I in mammalian cells increases following ionizing radiation (IR), indicating that Prx-I actively responds to IR-induced reactive oxygen species (ROS) and suggesting that Prx-I plays an important role in protecting cells from IR-induced death. To test this hypothesis, we suppressed the expression of Prx-I in SW480 cells by RNA interference. Our results show that IR induces the expression of Prx-I in SW480 cells in a dose- and time-dependent manner. The recombinant siRNA vector targeting Prx-I dramatically reduced the expression of Prx-I in SW480 cells. When Prx-I was knocked down in SW480 cells, the cells exhibited a decreased growth rate, a reduced antioxidant capability following IR and became more sensitive to IR-induced apoptosis. Together, our results demonstrate that Prx-I plays an important role in protecting cells from IR-induced cell death, which might be through scavenging IR-induced ROS in the cells. PMID- 16210787 TI - Exposure to a 50-hz magnetic field induces a circadian rhythm in 6 hydroxymelatonin sulfate excretion in mice. AB - The effect of magnetic field (MF) exposure on melatonin production was studied in female CD(2)F(1)(BALB/c x DBA/2) mice. The mice were exposed to a 50 Hz MF at 100 microT for 52 days and nocturnal urine was collected 1, 3, 7, 14, 16 and 23 days after the beginning of MF exposure. The animal room was illuminated for 12 h daily at 200 lux. To study the circadian rhythm of melatonin production, night and day samples of urine were collected once, at about 40 days after the beginning of MF exposure. Urinary 6-hydroxy melatonin sulfate (6-OHMS) was determined to assess melatonin production. The pineal glands were analyzed for melatonin content at the middle of the dark period. No statistically significant peak of melatonin was observed in either group. The light-regulated natural melatonin rhythm was absent in sham-exposed mice. The MF exposure caused a significant day-night difference in the 6-OHMS levels, but did not affect the total excretion of 6-OHMS during the 24-hour period. A possible interpretation of the findings is that MF exposure increases the sensitivity of the pineal gland to light in this strain normally insensitive to the circadian light variations. Further studies on interaction of light and MF exposure might help in understanding the inconsistencies of earlier research on MFs and melatonin. PMID- 16210788 TI - Homolytic cleavage of the O-glycoside bond in carbohydrates: a steady-state radiolysis study. AB - The formation of products resulting from the O-glycoside bond cleavage following radiolysis of aqueous solutions of methyl-alpha-D-glucopyranoside (I), 3-O-methyl alpha-D-glucopyranose (II), maltose, lactose, gentiobiose and cellobiose were studied. Radiation-induced destruction yields were also determined for dextran, laminarin and trimethylcelulose upon irradiation of their aqueous solutions. Oxygen, quinones and compounds capable of forming quinoid structures were found to inhibit radiation-induced homolytic destruction processes taking place in glycosides, di- and polysaccharides. The data obtained in this study enabled the authors to demonstrate an important role played by the fragmentation reaction of C-2 radicals which were generated from the starting substances in the formation of final radiolysis products. PMID- 16210789 TI - Repair kinetics of DNA-DSB induced by X-rays or carbon ions under oxic and hypoxic conditions. AB - We studied the relation between initial DNA double-strand breaks (DNA-DSB) and the rejoining kinetics of the strand breaks, as well as the OER (oxygen enhancement ratio) after low- and high-LET (linear energy transfer) radiations. CHO cells were exposed to 200 kVp X-rays or 80 keV/microm carbon ions under oxic and hypoxic conditions. DNA-DSB in the cells were analyzed by a static-field gel electrophoresis (SFGE). The kinetics of the rejoining could be described by a sum of fast and slow components. The initial released DNA after X-ray irradiation was higher for cells irradiated under an oxic condition than that under a hypoxic condition. The OER of DNA-DSB after X-ray irradiation was 5.7. This value decreased rapidly to be 3.4 with the fast component by 15 minutes. On the other hand, the OER of DNA-DSB after carbon ion irradiation was 2.2, and this value was not changed by rejoining incubation. The OER values for cell killing were 2.8 and 1.8 after X-ray and carbon ion irradiations, respectively. These values matched to the OER for DNA-DSB with complete rejoining. We conclude that the rejoining of DNA-DSB is an important factor in the mechanism of the oxygen effect. PMID- 16210790 TI - Estimation of yields of OH radicals in water irradiated by ionizing radiation. AB - The yield of OH radical induced by ionizing radiation was estimated by an empirical model; a prescribed diffusion model for a spur of single size applying to neutral water. Two representative spur distances were introduced, one for an incident primary charged particle and one for a representative secondary electron, to calculate chemical yields among active species in a spur. The total yield from the track was a combination of these primary and secondary yields. Two coefficients of this combination were the parameters of the present model. Based on an optimization of these parameters by existing experimental Fricke G-values, the present model estimates the yields of OH at the microsecond timescale after an irradiation, in a unified manner from electrons to heavy ions. The predicted yields of OH around the nanosecond timescale after an irradiation may be a relevant basis for a study on the mechanisms of radiation effects. This prediction by the present model was exemplified for electrons, photons and heavy ions (proton, helium, carbon, neon, argon and iron). PMID- 16210791 TI - Exploration of "over kill effect" of high-LET Ar- and Fe-ions by evaluating the fraction of non-hit cell and interphase death. AB - The reason why RBE for cell killing fell to less than unity (1.0) with very high LET heavy-ions ((40)Ar: 1,640 keV/microm; (56)Fe: 780, 1,200, 2,000 keV/microm) was explored by evaluating the fraction of non-hit cell (time-lapse observation) and cells undergoing interphase death (calculation based on our previous data). CHO cells were exposed to 4 Gy (30% survival dose) of Ar (1,640 keV/microm) or Fe ions (2,000 keV/microm). About 20% of all cells were judged to be non-hit, and about 10% cells survived radiation damage. About 70% cells died after dividing at least once (reproductive death) or without dividing (interphase death). RBE for reproductive (RBE[R]) and interphase (RBE[I]) death showed a similar LET dependence with maximum around 200 keV/microm. In this LET region, at 30% survival level, about 10% non-survivors underwent interphase death. The corresponding value for very high-LET Fe-ions (2,000 keV/microm) was not particularly high (approximately 15%), whereas that for X-rays was less than 3%. However, reproductive death (67%) predominated over interphase death (33%) even in regard to rather severely damaged cells (1% survival level) after exposure to Fe-ions (2,000 keV/microm). These indicate that interphase death is a type of cell death characteristic for the cells exposed to high-LET radiation and is not caused by "cellular over kill effect". Both NHF37 (non-hit fraction at 37% survival) and inactivation cross-section for reproductive death (sigma[R]) began to increase when LET exceeded 100 keV/microm. The exclusion of non-hit fraction in the calculation of surviving fraction partially prevented the fall of RBE[R] when LET exceeded 200 keV/microm. On the other hand, the mean number of lethal damage per unit dose (NLD/Gy) showed the same LET-dependent pattern as RBE[R]. These suggest that the increase in non-hit fraction and sigma[R] with an increasing LET is caused by enhanced clustering of ionization and DNA damage which lowers the energy efficiency for producing damage and RBE. PMID- 16210792 TI - Effects of 2450 MHz electromagnetic fields with a wide range of SARs on methylcholanthrene-induced transformation in C3H10T1/2 cells. AB - This study examined whether 2450 MHz continuous wave high frequency electromagnetic fields (HFEMF) could induce cancer-like changes in mouse C3H10T1/2 cells, and whether HFEMF could initiate malignant or synergistic transformation. Transformed foci, Type II and Type III, were independently counted as the experiment endpoint. The cells were exposed to HFEMF alone at a wide range of specific absorption rates (SARs) of 5 to 200 W/kg for 2 h and/or were treated with a known initiating chemical, methylcholanthrene (MC) (2.5 microg/ml). No significant differences were observed in the malignant transformation (Type II + Type III) frequency between the controls and HFEMF with or without 12-O-tetradecanoylphorbol-13-acetate (TPA) (0.5 ng/ml), a tumor promoter that could enhance transformation frequency initiated by MC in multistage carcinogenesis. However, the transformation frequency for HFEMF at SAR of more than 100 W/kg with MC or MC plus TPA was increased compared with MC alone or MC plus TPA. On the other hand, the corresponding heat groups (heat alone, heat + MC, and heat + MC + TPA) did not increase transformation compared with each control level in C3H10T1/2 cells. This result suggests that 2450 MHz HFEMF could not contribute to the initiation stage of tumor formation, but it may contribute to the promotion stage at the extremely high SAR (100 W/kg). PMID- 16210793 TI - Effects of hypoxic cell radiosensitizer doranidazole (PR-350) on the radioresponse of murine and human tumor cells in vitro and in vivo. AB - We have investigated the radiosensitizing effect of doranidazole, a hypoxic cells radiosensitizer, using SCCVII tumor cells of C3H mice and CFPAC-1 and MIA PaCa-2 human pancreatic tumor cells. The radiosensitivity of hypoxic SCCVII cells in vitro increased with 1 mM doranidazole by a factor of 1.34 and 1.68, when determined by clonogenic survival and micronucleus (MN) formation, respectively. The radiation-induced growth delay of SCCVII tumors was significantly enhanced and the TCD(50/120) was reduced by a factor of 1.33 when 200 mg/kg doranidazole was injected, i.v., 20 min prior to tumor irradiation. The in vivo-in vitro excision assay showed that radiosensitivity of SCCVII cells in vivo increased by a factor of 1.47 with 200 mg/kg doranidazole. The radiation-induced growth delay of CFPAC-1 xenografts in nude mice was significantly enhanced and the TCD(50/90) was reduced by a factor of 1.30 by 200 mg/kg doranidazole. On the other hand, 200 mg/kg of doranidazole exerted no influence on the radiation-induced growth delay in MIA PaCa-2 xenografts. The tumor oxygenation status, as determined with an oxygen sensitive needle probe and the immunohistological study using pimonidazole, indicated that MIA PaCa-2 tumors are better oxygenated than CFPAC-1 tumors. The relatively well-oxygenated status in MIA PaCa-2 tumor may account for the lack of radiosensitization by doranidazole. It is concluded that the magnitude of radiosensitization of tumors by doranidazole is dependent on the oxygenation status of the tumors and that doranidazole may be useful in increasing the response of hypoxic human pancreatic tumor to IORT. PMID- 16210794 TI - Effects of long-term oral administration of ketoprofen in clinically healthy beagle dogs. AB - To investigate the adverse effects of long-term administration of ketoprofen in dogs, ketoprofen (1 mg/kg) was administered to five clinically healthy beagle dogs (ketoprofen group) and gelatin capsules (control group) were administered to four clinically healthy beagle dogs for 30 days. We monitored the dogs through periodic physical examination, blood analyses, endoscopic examinations, fecal occult blood tests, renal function tests, urinalysis, urinary enzyme indices and cuticle bleeding time analysis. The lesions in the stomach, especially in the pyloric antrum, and fecal occult blood progressively worsened in the ketoprofen group. However, the differences between the ketoprofen group and the control group were not statistically significant. One dog in the ketoprofen group temporarily exhibited a decrease in renal plasma flow and two dogs exhibited enzymuria. However, these changes did not persist and the other examinations showed no significant difference between premedication and postmedication in the ketoprofen group. Therefore, the adverse effects of long-term administration of ketoprofen observed in this study were not clinically important in healthy dogs. Nevertheless, further investigation of adverse renal effects from long-term administration of ketoprofen is necessary in the dogs with subclinical renal disease. PMID- 16210795 TI - Control of the collagen fibril diameter in the equine superficial digital flexor tendon in horses by decorin. AB - The distribution pattern of collagen fibril diameter in the equine superficial digital flexor tendon (SDFT) is known to differ in central and peripheral areas of some regions. This study reports the essence of collagen fibril differences among different regions of the equine SDFT by transmission electron microscopic (TEM) and high-voltage electron microscopic observations and biochemical analysis. The distribution of large collagen fibrils increased but the density of collagen fibrils decreased from the proximal metacarpal region to the distal metacarpal region. Large collagen fibrils with an irregular cross-sectional profile were found more frequently in the middle metacarpal region than in other regions. Three-dimensional reconstruction of images of irregularly shaped collagen fibrils revealed that these fibrils are formed through fusion of small collagen fibrils with large ones. The amount of decorin, which reportedly inhibits the lateral fusion of collagen fibrils, decreased in the direction of the distal metacarpal region. On the other hand, the size of decorin gradually increased in the direction of the distal metacarpal region. These results suggest that regional differences in collagen fibril distribution and density of collagen fibrils in the SDFT are due, at least in part, to fusion of collagen fibrils and the concomitant regional differences in the amount and size of decorin. PMID- 16210796 TI - Canine tyrosine hydroxylase (TH) gene and dopamine beta -hydroxylase (DBH) gene: their sequences, genetic polymorphisms, and diversities among five different dog breeds. AB - Dopamine and noradrenaline are catecholamine neurotransmitters that are produced by biosynthetic enzymes such as tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DBH). As a first step to elucidate the genetic background of canine behavioral traits, we selected these genes as targets and sequenced these canine genes, and found that both were highly homologous with those of human beings. Then brain cDNAs derived from ten unrelated Beagles were used to search for polymorphisms in these genes. Four single nucleotide polymorphisms (SNPs) (C97T, G168A, G180A and C264T), one of which (C97T) will cause amino acid substitution in the TH gene, and two SNPs (C789A and A1819G), both of which will cause amino acid substitutions in the DBH gene were identified. The allelic frequencies among five dog breeds (47 Golden Retrievers, 41 Labrador Retrievers, 40 Malteses, 26 Miniature Schnauzers, and 39 Shibas) were examined and found to have significant variation between them with regards to all these SNPs, except for C97T in the TH gene and A1819G in the DBH gene. The polymorphisms of C97T and A1819G were found only in the Shiba. The present results suggest that the polymorphisms of the genes encoding catecholamine biosynthetic enzymes may become important markers for examining the genetic background of behavioral characteristics in dogs. PMID- 16210797 TI - Topographic analysis of flash visual evoked potentials in dogs. AB - Visual evoked potentials by flash stimulation (flash VEP) were analyzed in dogs using a topographic method. The flash VEP consisted of 3 positive (P1, P2 and P3) and 2 negative (N1 and N2) components by 150 msec after the flash stimuli. On the topographic mappings, a negative response area was observed in the frontal region of the scalp in the stimulated site followed by the shifting of the area to the contralateral frontal region and occipital region, during the first 100 msec. The negative response area in the frontal region in the stimulated site, contralateral frontal and temporal region, and occipital region were corresponded to N1, P2, and N2 on the flash VEP, respectively, according to their latencies. In the dogs with experimentally impaired the right lateral geniculate body, the latency of P2 was prolonged, and N2 and P3 were disappeared after the left eye stimulation. On the topographic mapping, only the early negative response area was detected on the stimulated site of the frontal region of the brain. Therefore, it is concluded that P1 and N1, P2, and N2 are referred to the retinal potentials, the potentials from the retina to the brainstem included the lateral geniculate body, and those from the brainstem to the visual cortex, respectively. PMID- 16210798 TI - Development of gender differences in DBA/2Cr mouse kidney morphology during maturation. AB - Although we recently clarified sex-based differences in mouse kidney morphology, the developmental processes responsible for these gender differences during maturation remain unclear. The present study analyzed the morphometry of kidneys from 20-, 30-, 50-, 60-, 70-, 90-, 120- and 150-day-old DBA/2Cr mice. Total kidney weight and ratio of kidney weight to body weight were larger in males than females beginning at 50 days of age. The percentage of renal corpuscles exhibiting a cuboidal parietal layer was higher in males than in females in the 70-day and older mice. The diameter of cortical renal corpuscles was larger in males than in females beginning on day 90. The number of proximal convoluted tubular cell nuclei was higher in females than in males from day 90 onward. Vacuolar structures in the proximal convoluted tubular epithelium became prominent in 70-day-old males. PAS-positive granules in the proximal straight tubular epithelium became prominent in females on day 50. This paper is the first to describe the development of gender differences in mouse kidney morphology. PMID- 16210799 TI - Plasma vitamin C concentration is not related to the incidence of ketosis in dairy cows during the early lactation period. AB - Many animals including cattle can synthesize vitamin C from glucose. The objective of this study was to investigate plasma vitamin C concentration in ketotic cows during the early lactation period because glucose supply for vitamin C synthesis might be limited in these cows. We measured plasma beta hydroxybutyrate (BHBA) concentration in 118 cows within 2 months after parturition. Subclinical/clinical ketosis was quantitatively determined using a plasma BHBA threshold of 1,200 microM. Plasma glucose concentration was lower in the ketotic cows than in the control cows but plasma vitamin C concentration did not differ between the control and the ketotic cows. Then we measured plasma vitamin C, BHBA and glucose levels in 7 cows during the periparturient period. Plasma BHBA increased and plasma glucose decreased after parturition but plasma vitamin C did not change. These results indicate that plasma vitamin C is not related to the incidence of ketosis in the early lactation period. We suggest that ketotic cows have the ability to produce vitamin C to meet its requirement in the early lactation period although glucose supply is not sufficient for milk production. Vitamin C synthesis is possibly given a high metabolic-priority for glucose in lactating cows. PMID- 16210800 TI - Caprine enteritis associated with Yersinia pseudotuberculosis infection. AB - Yersiniosis was prevalent among a caprine herd during the late autumn of 2003 in Iwate Prefecture, Japan. The disease affected 29 of about 100 lactating goats, but not dried or nonparous goats, mature male goats or kids. Four animals died within an epidemic period of 20 days. Affected animals developed decreased milk production with subsequent watery diarrhea, neutrophilia with increased band forms and multiple microabscesses characteristic of yersiniosis in the intestinal mucosa from the jejunum to caecum as well as in the mesenteric lymph nodes. Y. pseudotuberculosis serotype III was isolated from intestinal contents and mesenteric lymph nodes. The organism was also cultured from clinically normal dried animals. The outbreak might have been precipitated by multiple stress factors, such as lactation, cold weather, Corynebacterium pseudotuberculosis infection resulting in abscess formation and tapeworm and coccidium parasitisms. PMID- 16210801 TI - Dysfunction of erythropoietin-producing interstitial cells in the kidneys of ICR derived glomerulonephritis (ICGN) mice. AB - Anemia is a major secondary symptom in chronic renal disorder (CRD), but the precise cause of insufficient production of erythropoietin (EPO) remains unclear owing to the controversial localization of EPO-producing cells in the kidneys. The ICR-derived glomerulonephritis (ICGN) mouse, a new hereditary nephrotic mouse, is an appropriate model of anemia associated with CRD. By using an amplified in situ hybridization technique, we detected and counted the renal EPO producing cells under both normoxic and hypoxic conditions. The expression levels of renal EPO mRNA were quantified and oxygen gradients were also assessed immunohistochemically. Amplified in situ hybridization clarified that EPO producing cells were peritubular interstitial cells in the middle region of renal cortex in both ICR and ICGN mice. Hypoxia (7% O2) induced low oxygen tension in proximal tubular epithelial cells of renal cortex, and increased the expression of EPO mRNA and the number of EPO-producing cells in both ICR and ICGN mice. However, hypoxia did not increase the serum EPO levels in ICGN mice. The ICGN mouse is a good model for anemia associated with CRD, and the suppression of EPO protein production in the renal EPO-producing cells is considered to be a potential cause of anemia associated with CRD. PMID- 16210802 TI - Isolation of a human erythrocyte-adapted substrain of Babesia rodhaini and analysis of the merozoite surface protein gene sequences. AB - Babesia rodhaini is a rodent hemoparasite closely related to B. microti, the major causative agent of human babesiosis. We tested the infectivity of B. rodhaini for human erythrocytes by using the SCID mouse model in which the circulating erythrocytes were replaced with those of humans. Initially, parasites grew very poorly in the mouse model, but a variant capable of propagating in human erythrocytes emerged after an adaptation period of three weeks. In an attempt to identify parasite proteins involved in the alteration of host cell preference, an expression cDNA library of B. rodhaini was constructed and screened with immune mouse sera. Although we were able to obtain three merozoite surface protein (MSP) genes, sequences of these genes from both the parental strain and human erythrocyte-adapted substrain were identical. Our results suggest that B. rodhaini has potential ability to infect human erythrocytes, but development of this ability may not be brought about by an amino acid change in MSPs. PMID- 16210803 TI - Acceleration of wound healing by gelatin film dressings with epidermal growth factor. AB - We examined in vivo efficiency of a gelatin film sheet with epidermal growth factor (EGF) for a novel therapeutic device for cutaneous wound repair. NIH3T3 fibroblasts and PAM212 keratinocytes proliferated when the cells were incubated with the homogenate of EGF containing gelatin sheets, indicating that the gelatin sheet retained biologic activity of EGF in the process of sheet formation. To evaluate therapeutic effects, the EGF containing gelatin sheets or control sheets were applied onto partial-thickness skin wounds made on dorsa of hairless dogs. Wound closure in wounds treated with EGF containing gelatin sheets was accelerated when compared to that of wounds treated with control sheets, exhibiting earlier reepithelialization of the epidermis and highly regulated repair of extracellular matrix in the dermis. Therefore, we concluded that the gelatin film is a useful material to keep EGF stable and the EGF containing sheet has the ability to become an efficient therapeutic agent for superficial or deep partial-thickness wounds in the skin. PMID- 16210804 TI - Expression of nerve growth factor in itchy skins of atopic NC/NgaTnd mice. AB - Although the possible involvement of neurotrophic factors in itchy skins of atopic dermatitis has been predicted, the exact mechanism by which itch is induced remains unclear. Since nerve growth factor (NGF) has crucial effects on development and functions of sensory nerves, we determined production of NGF and extension of nerve fibers in skins of NC/NgaTnd mice with or without atopic dermatitis. NC/NgaTnd mice spontaneously develop atopic dermatitis-like skin lesions when they are raised in air-unregulated conventional circumstances. We quantified scratching behavior of NC/NgaTnd mice during the development of dermatitis using a novel analytical system and compared to clinical skin severity scores. A significant correlation between the severity of dermatitis and the increase in the number of scratches was identified, indicating that scratching behavior may associate with clinical skin conditions. NGF contents in the skin lesions of conventional NC/NgaTnd mice were significantly higher than those in SPF mice. Positive reactions for NGF were observed in keratinocytes and fibroblasts in affected skins of conventional NC/NgaTnd mice. Immunohistochemical analysis showed the extension of protein gene product 9.5-positive nerve fibers from the dermis toward the epidermis at the skin lesions. These results suggest that sensory nerves induced by NGF may contribute to development of itch, and that NGF produced at the affected site may provide abnormal skin sensitivity in atopic dermatitis. PMID- 16210805 TI - Molecular cloning and phylogenetic analysis of inflammatory cytokines of Camelidae (llama and camel). AB - We cloned, sequenced and analyzed the cDNAs encoding Camelidae inflammatory cytokines, including llama (lama glama) interleukin (IL)-1alpha, IL-1beta, IL-6, tumor necrosis factor (TNF)-alpha and camel (Camelus bactrianus) IL-6 and TNF alpha. The similarity levels of the deduced amino acid sequences of IL-1alpha, IL 1beta, IL-6 and TNF-alpha from llama (camel) to those from other mammalian species, ranged from 60.7% to 87.7%, 52.8% to 75.3%, 41.4% to 98.6%, and 72.9% to 99.6%, respectively. Phylogenetic analyses based on nucleic acid sequences showed that llama IL-1alpha, IL-1beta, IL-6 and TNF-alpha were more closely related to those of camel, pig, cattle, sheep and horse than to those of human, dog, cat, mouse and rat. PMID- 16210806 TI - Distribution of type VI collagen in the cartilaginous tissue of the proximal tibia in the domestic cat. AB - To investigate the distribution of the early stage chondrocytes during the formation and closure of epiphyseal growth plate (EGP) of the domestic cat, we examined the EGP of proximal tibiae by immunohistochemistry for type VI collagen. In the epiphyseal cartilage without the secondary ossification center (SOC) and EGP in newborn cats aged 1 and 10 days, type VI collagen-positive chondrocytes were located around the cartilage canals and articular surface. In the epiphyseal cartilage with the SOC and EGP in young cats aged 1 to 3 months, type VI collagen positive chondrocytes were located in the upper resting zone of the EGP, and then increased throughout the resting zone along with maturation. In the adult cats with the partially closed EGP, type VI collagen-positive chondrocytes were distributed throughout the remaining EGP. These findings indicate that the early stage chondrocytes characterized with type VI collagen are continuously located in the EGP during maturation. In addition, the increase of the early stage chondrocytes and the decrease of the reserve chondrocytes in the EGP along with maturation may cause the cessation of the longitudinal growth of the EGP, and finally bring about the EGP closure. PMID- 16210807 TI - Megaesophagus was complicated with Billroth I gastroduodenostomy in a cat. AB - A seven-year-old, female, domestic short hair cat was presented with a history of chronic anorexia. Radiographic examination revealed a large space-occupying calcified mass in the abdominal cavity. The mass was located in pylorus and did not extend into the duodenum and surrounding tissues. Billroth I gastroduodenostomy was conducted to remove the mass. Histopathological examination of the mass showed a lymphoma. Although Recovery following the operation was excellent, the patient showed intermittent vomiting unrelated to feeding. Radiographical examination revealed a megaesophagus, which was assumed to be a complication of the Billroth I procedure, since the condition was not observed before the procedure. PMID- 16210808 TI - Gastric adenocarcinoma with ossification in a ferret (Mustela putorius furo). AB - A 6-year-old female ferret had a firm mass 2 cm in diameter in the pyloric region of the stomach. Histopathologically, the mass was composed of neoplastic proliferation of well-differentiated epithelial cells, showing tubular or glandular growth patterns. Osseous metaplastic foci were often found in the tumor. Tumor cells showed a positive reaction for immunohistochemistry against bone morphogenetic protein-6, an osteogenic factor. A diagnosis of gastric adenocarcinoma with ossification was made. PMID- 16210809 TI - A canine pemphigus foliaceus case showing parallel relationship of disease activity and titer of serum anti-keratinocyte cell surface antibodies. AB - A seven-year-old, spayed female mongrel dog was diagnosed as pemphigus foliaceus (PF) by clinical, histopathological and immunopathological observations. Serum antibodies against the cell surface of keratinocytes in the dog were detected by indirect immunofluorescence (IIF) using cryosectioned bovine esophagus as well as living cultured-canine keratinocytes as the substrates. When we compared the titers of IIF on bovine esophagus with its disease activity, the IIF titers reflected the disease activity throughout the time course. Our findings will suggest that sequential titration of serum antibodies by IIF will be useful for monitoring the serological disease activity in canine PF. PMID- 16210810 TI - Field basis evaluation of Eimeria necatrix-specific enzyme-linked immunosorbent assay (ELISA) for its utility in detecting antibodies elicited by vaccination in chickens. AB - Eimeria necatrix-specific ELISA, using a recombinant antigen (the cDNA-clone NP19 expressing protein), was utilized to detect antibodies against E. necatrix in breeder pullet flocks that had previously received an attenuated live vaccine to E. necatrix. Vaccinated flocks were discriminated significantly from non vaccinated flocks by their antibody titers and antibody positive rates at 30-55 days post-vaccination. In addition, E. necatrix-oocysts were confirmed in fecal samples of vaccinated flocks using PCR in the case where the antibody positive rates rose. These findings implied that the vaccination prompted repeated infections, and consequently the chickens generated antibodies and secured their protection against virulent field-E. necatrix. Therefore, the ELISA was suggested to be a useful tool to estimate the immune state of chickens as a result of vaccination with a live E. necatrix-vaccine. PMID- 16210811 TI - Bacteriological survey of feces from feral pigeons in Japan. AB - Some public areas in Japan such as parks and gardens can be highly contaminated with pigeon feces. We examined levels of four bacterial contaminations in fecal samples from feral pigeons in 7 prefectures. We isolated Salmonella Typhimurium and S. Cerro from 17 (3.9%) of 436 samples, as well as Mycobacterium spp. including M. avium-intracellulare complex from 29 (19.0%) of 153 samples. The polymerase chain reaction detected Chlamydia psittaci and C. pecorum in 106 (22.9%) of 463 samples, but E. coli O-157 was not isolated from any of the samples. Our results indicate that pigeon feces are a source of several zoonotic agents for birds, animals and humans. PMID- 16210812 TI - Different effects on the inflammatory lesions in the lacrimal and salivary glands after neonatal thymectomy in IQI/Jic mice, a model for Sjogren's syndrome. AB - Thymectomy on day 3 after birth (D3Tx) is understood to eliminate CD4+CD25+ regulatory T cells (Treg) from the peripheral T cell repertoire in rodents, leading to the activation of autoreactive T cells. Herein, D3Tx was performed in IQI/Jic mice, a model for Sjogren's syndrome characterized by autoimmune infiltrations into the lacrimal and salivary glands. At the age of 16 weeks, very severe lesions were observed in lacrimal tissues from thymectomized mice, suggesting that Treg preserve their immunoregulatory function in young IQI/Jic mice. In contrast, salivary lesions were comparable in the D3Tx and control groups, raising the possibility that either salivary-specific Treg escaped elimination in thymectomized mice or spontaneous lesions in IQI/Jic mice developed independently of the tolerance through Treg. PMID- 16210813 TI - Characteristics of Rathke's cleft cyst in MR imaging. AB - PURPOSE: Symptoms, macroscopic appearances and microscopic findings of Rathke's cleft cysts with magnetic resonance (MR) imaging. PATIENTS AND METHODS: We analyzed the data from 31 patients with pathologically confirmed Rathke's cleft cysts. MR appearances were evaluated on T1WI, T2WI and contrast T1WI. Symptoms, macroscopic appearances and pathological findings were obtained from available medical records. We analyzed the images according to the following criteria: 1. findings on the location and shape of the lesions and form of the lesional wall; 2. the relationship between the maximum diameter of the lesions and symptoms; 3. the relationship between MR and macroscopic appearance; 4. the sites of adjacent contrast enhancement. RESULTS: The lesions were located mostly in both the intrasellar and suprasellar regions for a total of 87%. All lesions revealed either an oval or dumbbell shape with a smooth lesional wall. When correlated with physical symptoms, asymptomatic cases were associated with smaller lesions, while visual disturbances and dizziness were associated with relatively larger lesions. MR lesion signal intensity was related to the content of macroscopic appearance to some degree: the selected lesions showed shortening of T1 and T2 relaxation times in relation to the increase in protein concentration. This should have been macroscopically reflected in the color and turbidity of the liquid within the cyst. Adjacent contrast enhancement around the lesion was found at various sites, but anterior enhancement was most frequent. Circumferential enhancement was revealed to be derived from inflammatory changes. CONCLUSION: Rathke's cleft cyst exhibits a varied MR signal. It may be difficult to differentiate from craniopharyngioma from the intensity alone. PMID- 16210814 TI - Staging of malignant lymphoma with three-station black-blood fast short-inversion time inversion recovery (STIR). AB - PURPOSE: The purpose of this study was to assess the usefulness of three-station black-blood fast short-inversion time inversion recovery (STIR) imaging in detecting and staging malignant lymphoma. METHODS: Seventeen patients with malignant lymphoma were examined with a 1.5T imager. The findings and stagings determined with three-station black-blood fast STIR imaging were compared with reference standards (e.g., computed tomography [CT] findings and clinical stagings). RESULTS: Three-station black-blood fast STIR imaging provided a fat suppressed T2-weighted imaging contrast with fewer flow artifacts and revealed nodal involvement as well as bone marrow and spleen involvement to an extent comparable with CT. Especially notable was the excellent specificity (94%) of this imaging technique. Regarding disease staging, significant agreement was observed between clinical staging (k=0.60) and staging as evaluated by three station black-blood fast STIR, although the detection of lymphadenopathy in the thorax was relatively poor. The average time required for this imaging was approximately 30 min. CONCLUSION: Three-station black-blood fast STIR MR imaging may be useful as a staging tool for malignant lymphoma because this imaging technique reveals lymphoma lesions, which determine the staging, without radiation exposure or the use of contrast agents. PMID- 16210815 TI - Feasibility of internally referenced brain temperature imaging with a metabolite signal. AB - The feasibility of using a metabolite signal as an internal reference for self referenced temperature distribution measurement was examined. Line scan echo planar spectroscopic imaging (LSEPSI) was applied to obtain quick multi-voxel spectroscopic measurements and to avoid possible spectral degradation from motion. Temperature distribution in a rabbit brain in vivo was successfully visualized by means of the chemical shift of water, which was measured by using naturally abundant (up to 10 mM) N-acetyl-aspartate (NAA) as the reference signal. Unlike the phase-mapping approach, this technique does not require a pixel-by-pixel subtraction. Therefore, in theory, it is more resistant to inter scan motion or changes in susceptibility. The spatial and temporal resolutions of this technique are 1.5 cm3 and 4.5 min. A higher signal-to-noise ratio and optimization of the water and outer-volume suppression capabilities will be required to further enhance the temperature-mapping capabilities. PMID- 16210816 TI - All-phase MR angiography using independent component analysis of dynamic contrast enhanced MRI time series: phi-MRA. AB - Dynamic contrast enhanced magnetic resonance imaging (dynamic MRI) represents a MRI version of non-diffusible tracer methods, the main clinical use of which is the physiological construction of what is conventionally referred to as perfusion images. The raw data utilized for constructing MRI perfusion images are time series of pixel signal alterations associated with the passage of a gadolinium containing contrast agent. Such time series are highly compatible with independent component analysis (ICA), a novel statistical signal processing technique capable of effectively separating a single mixture of multiple signals into their original independent source signals (blind separation). Accordingly, we applied ICA to dynamic MRI time series. The technique was found to be powerful, allowing for hitherto unobtainable assessment of regional cerebral hemodynamics in vivo. PMID- 16210817 TI - Feasibility of a curvature-based enhanced display system for detecting cerebral aneurysms in MR angiography. AB - PURPOSE: To assess the feasibility of a curvature-based enhanced display system for detecting cerebral aneurysms in MR angiography. METHODS: MR angiography studies of 18 patients (eight male and 10 female, average age 65.7, age range 50 to 75 years old) with 23 known aneurysms were evaluated with a curvature-based display system. The two curvature features-the volumetric shape index and curvedness values-were calculated at each voxel. These were displayed independently on a workstation, overlaid on volume-rendered images. Two neuroradiologists evaluated the images for visibility and diagnosis of the cerebral aneurysms. The diagnostic results were compared with the original reports. RESULTS: The calculation time for each curvature index was 30 to 40 s for 120 to 140 slices of original MR angiography data. Shape index images emphasized smooth and round aneurysms more than aneurysms with irregular surfaces. Curvedness images revealed aneurysms well when the aneurysms had diameters that differed from those of the surrounding vessels. The computer assisted-detection method detected 24 aneurysms, three of which were not pointed out in the initial report. CONCLUSION: Our results show that the curvature-based display system we have developed is feasible and that it may help to detect small aneurysms that are prone to be overlooked in routine readings. PMID- 16210818 TI - 3T MR for clinical use: update. AB - Clinical experience with 3-tesla MRI for body applications has only a short history; to date, it has no proven practical advantage over 1.5-tesla or less powerful systems. However, the theoretical advantage of higher field strength which includes a higher signal-to-noise ratio with identical scan parameters, higher special resolution available within clinically acceptable scan time, and better spectral resolution-can contribute to the clinical outcome. This paper seeks to demonstrate some of the clinical advantages of the 3-tesla system through preliminary experiences as well as the potential advantages of a 3-tesla system when combined with much of the hardware and software that is clinically accepted in the 1.5-tesla world. PMID- 16210819 TI - Feasibility of quantitative intrahepatic lipid imaging applied to the magnetic resonance dual gradient echo sequence. AB - The quantification of intrahepatic lipids (IHLs) has attracted a great deal of interest pertaining to diagnostic treatment of type 2 diabetes mellitus. We report on an innovative approach to visualizing IHLs quantitatively by creating the best mix of the advantages of proton magnetic resonance spectroscopy (MRS) and the gradient echo (GRE) sequence on a 1.5T MR system. Proton MRS is considered to have the best precision and reliability; however, measuring a single voxel is time-consuming. On the other hand, the GRE sequence can provide information on IHLs, at least not quantitative, in a very short time. IHL images can be created from the correlation between the relative content of IHLs obtained with proton MRS and the signal intensity of GRE images with a very high correlation coefficient (R=0.992). Our approach holds great potential for quantifying fat accumulation in every possible tissue quickly and precisely. PMID- 16210820 TI - MR microscopy of the articular cartilage with a 1.0T permanent magnet portable MR system: preliminary results. AB - The present study was designed to evaluate the use of a 1.0T portable permanent magnet MR system in obtaining microscopic MR images of the hyaline cartilage in vitro. A clear laminar appearance was demonstrated with this system. In addition, it was possible to demonstrate cartilage surface irregularity, a decrease in cartilage thickness, and T2 alteration by proteoglycan depletion following up to 12 hours of trypsin treatment. In summary, the portable MR system is useful for investigating cartilage in vitro. PMID- 16210821 TI - MR imaging of hepatocellular carcinoma arising in genetic hemochromatosis. AB - A case of a 51-year-old male with previously unsuspected genetic hemochromatosis (GH) associated with hepatocellular carcinoma (HCC), for which MR imaging was useful in diagnosis, is presented. Contrast-enhanced ultrasonography depicted a mass in segment 8 of the liver and a liver tumor was suspected. The mass could not be revealed on unenhanced or enhanced CT examination. MR imaging with unenhanced T1-weighted FLASH and T2-weighted gradient echo sequences clearly depicted the tumor; the tumor appeared hyperintense against a background of low hepatic signal intensity caused by iron overload. The iron-overloaded liver provided a natural source of paramagnetic contrast for detection of HCC. Studies are required to further assess the accuracy of these sequences in detection of HCC in patients with GH. PMID- 16210822 TI - First-pass myocardial perfusion defect and delayed contrast enhancement in hypertrophic cardiomyopathy assessed with MRI. AB - BACKGROUND: Gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) enhanced magnetic resonance imaging (MRI) is known to be useful for detecting myocardial injury. In this study, we used first-pass myocardial perfusion and delayed contrast-enhanced MRI to determine whether an abnormal signal intensity was related to the left ventricular regional contractile function in patients with hypertrophic cardiomyopathy (HCM). MATERIALS AND METHODS: Twelve patients with HCM participated in this study. Four short axial cine images of the left ventricle were acquired. Subsequently, first-pass myocardial perfusion images during the first passage of Gd-DTPA (0.1 mmol/kg), and delayed contrast-enhanced images after a 15-min delay, were acquired in the same orientation as cine imaging. Each image was divided into eight blocks and a total of 384 blocks were analyzed. RESULTS: First-pass myocardial perfusion defects (PD) were detected in nine patients with an average of 11.5+/-11 blocks. Delayed contrast enhancement (DE) was detected in 11 patients with an average of 11.5+/-10 blocks. Mean wall thickness in PD blocks (16.7+/-4.7 mm) was larger than that in normal perfusion blocks (13.6+/-3.9 mm, p<0.001). Mean wall thickness in DE blocks (16.9+/-4.9 mm) was larger than that in normal enhanced blocks (13.4+/-3.6 mm, p<0.001). PD were located at almost the same site as DE, but DE areas were larger than PD areas (p=0.0021). Mean percent wall thickening of blocks with PD (63.1+/-44.7%, p<0.0001) and blocks with DE (75.2+/-81.5%, p<0.01) was lower than that in blocks with neither PD nor DE (103.5+/-66.0%). Significant correlations were found between percent wall thickening and percent PD (r=0.46, p<0.0001) and between percent wall thickening and percent DE (r=0.54, p<0.0001). CONCLUSION: Abnormal signal intensity from first-pass myocardial perfusion and delayed contrast enhanced MRI are closely related to left ventricular regional contractile function. PMID- 16210823 TI - Artifacts of vena cava filters ex vivo on MR angiography. AB - We evaluated magnetic susceptibility artifacts of nine types of vena cava filters in MR angiography (MRA) at 1.0T ex vivo in order to assess the filters' compatibility with MRA. Each filter (tulip filter, tulip MReye filter, stainless Greenfield filter, titanium Greenfield filter, TrapEase filter, Simon filter, LGM Vena-Tech filter, Antheor temporary filter, and Bird's nest filter) was inserted into an acrylic tube (20 or 25 mm in diameter, 15 or 30 cm in length). Gd-DTPA was poured into each tube at a concentration of 1/500 and each was placed in a water-filled container for imaging. We evaluated artifacts of the filters according to the following criteria: signal void beyond the tube, 3+; signal void within the tube but at more than one-half the diameter of the tube, 2+; and signal void within the tube but at less than one-half the diameter of the tube, 1+. We evaluated artifacts originating at the tip, intermediate portion, and distal end of the filters. We judged the artifacts as follows: tulip (3+, 3+, 3+); tulip MReye (2+, 1+, 1+); stainless Greenfield (2+, 1+, 2+); titanium Greenfield (1+, 1+, 1+); TrapEase (1+, 2+, 1+); Simon (2+, 2+, 1+); LGM (2+, 2+, 1+); Antheor (2+, 2+, 2+); and Bird's nest (3+, 3+, 3+). The numbers in parentheses refer to the degree of signal void at the tip, intermediate portion, and distal end of the filter, respectively. The tulip filter and Bird's nest filter made of 304 stainless steel caused extensive signal voids beyond the areas defined by the filters. The signal voids in the remaining seven filters were limited to within the tube. We concluded that seven of the nine filters were compatible with MRA ex vivo. PMID- 16210824 TI - Measurement of oxidative stress in the rodent brain using computerized electron spin resonance tomography. AB - The strategy of this study was to improve the electronic spin resonance (ESR) application used to detect free radical-induced oxidative stress in animal models. We have developed an in vivo ESR imaging system with high-quality ESR computed tomography (CT) images by using a nitroxyl spin probe--BBB-permeable, 3 methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine-1-yloxy (MC-PROXYL)--in living small animals. We first measured the distribution of MC-PROXYL in the head region of a living mouse by using ESR-CT imaging after treatment with MC-PROXYL. In the ESR-CT experiments, it was clearly observed that MC-PROXYL was well distributed in the brain of head region of a living mouse. The ESR-CT images taken after treatment with MC-PROXYL demonstrate that the decay of MC-PROXYL in the isolated brain of a spontaneously hypertensive rat (SHR) was more rapid than that in a Wister Kyoto rat. ESR-CT analysis revealed that the region of rapid decay might be located in the cerebral cortex of the isolated brain of an SHR. These results suggest that the present study of ESR-CT imaging would be a useful tool for monitoring and detecting the locations of oxidative stress in the brains of rodent animal models. PMID- 16210825 TI - Autoregressive moving average (ARMA) model applied to quantification of cerebral blood flow using dynamic susceptibility contrast-enhanced magnetic resonance imaging. AB - PURPOSE: To investigate the feasibility of the autoregressive moving average (ARMA) model for quantification of cerebral blood flow (CBF) with dynamic susceptibility contrast-enhanced magnetic resonance imaging (DSC-MRI) in comparison with deconvolution analysis based on singular value decomposition (DA SVD). METHODS: Using computer simulations, we generated a time-dependent concentration of the contrast agent in the volume of interest (VOI) from the arterial input function (AIF) modeled as a gamma-variate function under various CBFs, cerebral blood volumes and signal-to-noise ratios (SNRs) for three different types of residue function (exponential, triangular, and box-shaped). We also considered the effects of delay and dispersion in AIF. The ARMA model and DA SVD were used to estimate CBF values from the simulated concentration-time curves in the VOI and AIFs, and the estimated values were compared with the assumed values. RESULTS: We found that the CBF value estimated by the ARMA model was more sensitive to the SNR and the delay in AIF than that obtained by DA-SVD. Although the ARMA model considerably overestimated CBF at low SNRs, it estimated the CBF more accurately than did DA-SVD at high SNRs for the exponential or triangular residue function. CONCLUSION: We believe this study will contribute to an understanding of the usefulness and limitations of the ARMA model when applied to quantification of CBF with DSC-MRI. PMID- 16210826 TI - Development of intraarterial contrast-enhanced 2D MRDSA with a 0.3 tesla open MRI system. AB - PURPOSE: The purpose of this study was to develop a new technique for a high temporal resolution two-dimensional MR digital subtraction angiography (2D MRDSA) sequence under intraarterial injection of contrast material to permit the visualization of vascular anatomy and hemodynamics. METHODS: 2D MRDSA was imaged on a 0.3T open MR scanner with a T(1)-weighted fast gradient echo sequence. The phantom study examined vials containing gadolinium (Gd) solutions ranging in concentration from 0.5 mmol/L to 100 mmol/L. Repetition time and echo time were fixed at minimal values in order to achieve high temporal resolution, and only the flip angle was changed in 10-degree increments between 10 and 90 degrees. The in vivo study examined a brachial artery of a human volunteer. MRDSA images were acquired continuously during intraarterial injections of Gd solutions ranging in concentration from 0.5 mmol/L to 100 mmol/L. The subtracted images were displayed on the monitor in real time at a frame rate of one frame per second and evaluated to determine the optimal concentration of contrast material. RESULTS: In the phantom study, a 10-mmol/L Gd concentration with a flip angle of 50 degrees -90 degrees and a 25-mmol/L Gd concentration with a flip angle of 60 degrees -90 degrees showed high signal-to-noise ratios. In the human brachial artery experiment, the forearm arteries were well visualized when solutions of 5-50 mmol/L Gd concentration were used. The 10- and 25-mmol/L Gd concentrations were considered optimal. The palmar digital arteries were also visualized. Higher Gd concentrations showed a paradoxical signal increase when diluted by blood. CONCLUSION: We successfully developed an intraarterial contrast-enhanced 2D MRDSA sequence. With appropriate settings of imaging parameters and Gd concentrations, we obtained acceptable vessel visualization in the human study. The low Gd concentration for optimal visualization permits repeated intraarterial injections. This technique can be a useful tool for investigating the vascular anatomy and hemodynamics required for MR-guided vascular interventions. PMID- 16210827 TI - MRI findings of an ovarian dermoid cyst with malignant transformation. AB - The ovarian dermoid cyst is the most common ovarian tumor. However, malignant transformation developing from a dermoid cyst is very rare. Because of this rarity, few reports exist of preoperative diagnosis of this tumor by MRI. We report MRI findings from a 69-year-old patient with a malignant transformation (squamous cell carcinoma with sarcomatoid features) in a right ovarian dermoid cyst. PMID- 16210828 TI - The use of three-dimensional ultrasound micro-imaging to monitor prostate tumor development in a transgenic prostate cancer mouse model. AB - Longitudinal studies of mouse cancer models required large cohorts since autopsy was the only reliable method to evaluate treatment efficacy. This paper reports the use of high-resolution three-dimensional ultrasound micro-imaging to monitor prostate tumor development in genetically engineered mice. Twenty-nine genetically engineered prostate cancer mice, including castrated and uncastrated mice, were imaged by three-dimensional ultrasound. Qualitative comparisons of three-dimensional ultrasound images with histology sections of prostate tumors demonstrate the ability of ultrasound to accurately depict the size and shape of malignant masses in live mice. The correlation coefficient of tumor diameter measurements performed in vivo with three-dimensional ultrasound and at autopsy was 0.997. Prospective tumor detection sensitivity and specificity were 91.7% and 100%. Representative exponential growth curves constructed via longitudinal ultrasound imaging indicated diameter doubling times from 10 to 37 days for four prostate tumors during an initial period of rapid progression. Three-dimensional ultrasound will likely become the micro-imaging modality most readily adopted for mouse pre-clinical trial studies. PMID- 16210829 TI - Neuromagnetic evidence that gingiva area is adjacent to tongue area in human primary somatosensory cortex. AB - The somatotopic organization of the human primary somatosensory (SI) area in the cerebral cortex has been intensively studied for the hand, lip, and tongue, but little is known about the gingiva. Penfield concluded that the gingival SI area was above the tongue area, as shown in his famous homunculus map. However, our recent study suggested that the lingual gingiva area was not so different to the tongue area. To delineate the fine SI somatotopy of the gingiva area, evoked magnetic fields were measured in 6 healthy subjects for the stimulus of the anterior or posterior and upper or lower parts of the lip, buccal and lingual gingiva, and tongue. Source position was estimated by a current dipole model at the first peak of the posterior-oriented current in a total of 12 cerebral hemispheres contralateral to the stimulation side. No significant difference was found between the positions of anterior and posterior or upper and lower parts of each structure. Both buccal and lingual gingiva areas were localized adjacent to the tongue area, but significantly lower than the lip area. We believe that the fine SI somatotopy of the human oral structures should be reconsidered. PMID- 16210830 TI - Clinical application of acceleration sensor to detect the swing phase of stroke gait in functional electrical stimulation. AB - Functional electrical stimulation (FES) can improve the gait of stroke patients by stimulating the peroneal nerve in the swing phase of the affected leg, causing dorsiflexion of the foot that allows the toes to clear the ground. A sensor can trigger the electrical stimulation automatically during the stroke gait. We previously used a heel sensor system, which detects the contact pressure of the heel, in FES to correct foot drop gait. However, the heel sensor has disadvantages in cosmetics and durability. Therefore, we have replaced the heel sensor with an acceleration sensor that can detect the swing phase based on the acceleration speed of the affected leg, using a machine learning technique (Neural Network). We have used a signal for heel contact in a gait using the heel sensor before training with the Neural Network. The accuracy of the Neural Network detector was compared with a swing phase detector based on the heel sensor. The Neural Network detector was able to detect similarly the swing phase in the heel sensor. The largest difference in timing of the swing phase was less than 60 milliseconds in normal subjects and 80 milliseconds in stroke patients. We were able to correct foot drop gait using FES with an acceleration sensor and Neural Network detector. The present results indicate that an acceleration sensor positioned on the thigh, which is cosmetically preferable to systems in which the sensor is farther from the entry point of the electrodes, is useful for correction of stroke gait using FES. PMID- 16210831 TI - Trapidil, an inhibitor for phosphodiesterase and platelet-derived-growth factor, ameliorates corrosive esophageal burn in rats. AB - Corrosive esophageal burn is a common health problem in the pediatric age group and causes serious esophageal injuries. The medical treatment in acute phase of corrosive esophageal injury is of particular importance for prevention of esophageal stricture. We therefore aimed to investigate the possible beneficial effect of trapidil (triazolopyrimidine), an inhibitor for phosphodiesterase and platelet-derived-growth-factor, during acute phase of esophageal corrosive injury. Wistar albino rats were randomly allocated to untreated, treated, and sham-operated groups (n = 10 for each group). Corrosive esophageal burn was generated with 10% NaOH solution. The rats were left untreated (untreated group) or treated with trapidil as a single dose of 40 mg/kg intraperitoneally after one hour of the injury (treated group). Abdominal esophageal segment was isolated and tied in sham-control group. The studied esophageal segment was removed from each animal after 24 hours. Malondialdehyde (MDA) and nitric oxide (NO) levels were measured in the esophageal tissues. The ulcer depth was graded by histopathologic examination. MDA and NO levels were significantly higher in the untreated group than in the treated group. Namely, trapidil treatment significantly decreased MDA and NO levels in the injured tissues, the levels of which are similar to those in the tissues of control animals. The grades of ulcer depth were significantly improved in the treated group. These results indicate that the reactive oxygen radicals increase in the early phase of corrosive esophagitis and cause tissue damage. We suggest that trapidil treatment may be useful in acute phase of corrosive esophageal injury. PMID- 16210833 TI - Development of a food-frequency questionnaire to measure the dietary calcium intake of adult Japanese women. AB - Assessing calcium intake is necessary for epidemiological studies on osteoporosis. This study was aimed at developing a valid food frequency questionnaire (FFQ) to estimate the dietary calcium intake of Japanese people. The FFQ contains 26 items, representing common sources of calcium. The respondents, 74 women aged 15 - 79 years living in Fukui city, whose dietary calcium intake was estimated at 638 +/- 306 (S.D.) mg based on a one-day weighted food record kept during the National Nutrition Survey of 1996, were asked to indicate their consumption frequency and usual portion size (large, medium, small) for each food item. Trained dieticians interviewed the subjects to confirm their answers to the FFQs. The mean calcium intake per day according to the FFQ (655 +/- 222 mg) did not differ significantly from that according to the one-day food record. The coefficient of correlation between these methods was 0.512, a statistically significant value. The calcium intake from dairy products showed a good correlation, r = 0.717. The coefficient of correlation between the values estimated based on two FFQ filled out three days apart was 0.900, showing good test-retest reliability. Therefore, this FFQ is valid and reliable in estimating dietary calcium intake for epidemiological studies. PMID- 16210832 TI - A new preadipocyte cell line, AP-18, established from adult mouse adipose tissue. AB - Establishing preadipocyte cell lines from mature adipose tissues could help lead to a better understanding of adipogenesis. We have established a unique preadipocyte cell line, AP-18, derived from the subepidermal layer of ear skin from an adult C3H/HeM mouse. AP-18 cells exhibit fibroblast-like morphology, slow growth, and contact inhibition. The doubling time of AP-18 cells is 50-60 h, which is about 2-fold longer than that of well-known 3T3-L1 cells derived from mouse embryos. A small population of AP-18 cells spontaneously differentiates into adipocytes by 8 days after confluence, as judged by the accumulation of triglyceride droplets. Treatment of confluent AP-18 preadipocytes with adipogenic agents, containing dexamethasone, 3-methyl-1-isobutylxanthine, and insulin, increased triglyceride contents about 5-fold compared to the contents in untreated cells. We also analyzed mRNA expression profiles for key transcription factors involved in adipocyte differentiation, peroxisome proliferator-activated receptor (PPAR)gamma and the CCAAT/enhancer binding protein (C/EBP) family, and for differentiation markers, aP2, adipocyte-specific fatty acid-binding protein and adipsin, adipocyte-specific serine protease. AP-18 preadipocytes express mRNAs for C/EBPbeta, C/EBPalpha, PPARgamma, and aP2 before differentiation, but not adipsin mRNA. Expression of aP2 mRNA was increased in fully differentiated AP 18 cells. Likewise, expression of adipsin mRNA was increased after induced differentiation of AP-18 cells and reached the highest level in fully differentiated adipocytes. Thus, differentiation of AP-18 cells is associated with the increased expression of aP2 and adipsin mRNAs. The newly established AP 18 cell line provides a useful model for investigating adipocyte differentiation and adipogenesis. PMID- 16210834 TI - Mood scores in relation to hormone replacement therapies during menopause: a prospective randomized trial. AB - There is lack of studies in literature about the long-term effects of hormone replacement therapies and cholesterol levels on mood scores in menopause. In the present study we have investigated whether serum lipid levels affect mood scores in menopause and evaluated the long-term effects of the combined hormone replacement regimens (HRT) on depressive symptoms in postmenopausal women. In this prospective-randomized, placebo-controlled, double-blind study, 286 women in menopause were divided into four groups according to therapeutic regimens they received; 1) Conjugated equine estrogen (CEE) of 0.625 mg plus medroxyprogesterone acetate (MPA) of 2.5 mg (n = 79), 2) CEE (0.625 mg) plus MPA of 5 mg (n = 77), 3) tibolone of 2.5 mg (a selective tissue estrogenic activity regulator) (n = 76), and 4) Calcium (Ca) of 1,000 mg (n = 54). Beck Depression Inventory (BDI), and serum levels of lipoprotein lipids were assessed before and after 12-months of treatment with oral continuous HRT and Ca supplementation. BDI scores in the study groups were not correlated with lipid profiles. We compared two subgroups of patients with initial BDI scores 0-14 (normal mood scores) in order to asses for the possible relation between the lipid profile and mood. Following treatment, first subgroup had increased scores to 15-30 (mildly depressed women, n = 27) and the second subgroup preserved BDI scores of 0-14 (normal mood scores, n = 23). Serum levels of total cholesterol, high-density lipoprotein, low-density lipoprotein and body mass index were found to be similar between these two groups. BDI scores decreased significantly in all HRT groups after 12 months of treatment, compared to Ca group (p < 0.05). We did not observe any correlation between BDI scores and lipid profiles before and following continuous HRT or Ca supplementation. Continuous combined hormone replacement regimens, CEE + MPA and tibolone, have superior long-term effects on mood scores in menopause and should be considered during the decision process for use of HRT due to menopausal symptoms. PMID- 16210835 TI - Midterm results with the use of polycarbonate urethane heterografts for dialysis access. AB - Dialysis access surgery is currently one of the most common vascular operations as the dialysis patient population increases. Although autogenous arteriovenous fistulae stand as the preferred method of access due to excellent patency and lower complication rates, grafts may provide alternative approaches when an autogenous fistula is impractical usually due to depleted veins. We aimed in this study to assess early and midterm outcome with the use of Expedial heterografts (LeMaitre Vascular Ltd., Wrexham, UK) for angioaccess between October 2000 and December 2004. Patients were prospectively followed for pre- and peri-operative course with 1st, 6th and 12th month outpatient controls for patency and complication rates. A total of 34 heterografts were implanted in 30 patients (22 males and 8 females; median age 60.6 years, range 32 - 70). The mean followup for 30 patients were 16.3 +/- 6.1 (range 4 - 30) months. 28 grafts (82.4%) were placed in forearm position. Primary and secondary patencies were found as 81% and 94%, respectively (p = 0.15), with the median censored primary and secondary patencies of 27 +/- 1.7 months (95% CI, 23.38 to 30.67) and 30.42 +/- 1.1 months (95% CI, 28.30 to 32.54), respectively. Only presence of diabetes was found as a significant risk factor in the regression model (p = 0.01). We suggest that the polycarbonate urethane heterografts with acceptable patency/complication rates and shorter maturation times for dialysis access serve an alternative for these patients. PMID- 16210836 TI - Spinal anesthesia in a patient with severe thoracolumbar kyphoscoliosis. AB - Patients with spine abnormalities, present unique challenges to the health care provider responsible for administrating sedation and anesthesia during surgical and technical procedures. Spinal deformities may cause difficulties with both tracheal intubation and regional anesthesia. This report describes the anesthetic management for two urological operations that were performed in a patient with extremely severe thoracolumbar kyphoscoliosis. After examining the risk factors, spinal block by injecting single dose local anesthetic solution to the intratechal space was chosen to provide anesthesia. It has been suggested that hyperbaric solution, which is of high density compared with cerebrospinal fluid, can safely produce blocks for many operations under spinal anesthesia. In the first procedure, intrathecal injection of 6 mg hyperbaric bupivacaine, a local anesthetic solution (1.2 ml total volume), resulted in inadequate motor and sensory blockade, but the successful motor and sensory blockade at the level of Th10 was achieved in a second attempt with 6.25 mg hypobaric bupivacaine (2 ml). Because of this unexpected effect of local anesthetic solution, in the second operation, the technique was changed to intrathecal injection of 12.5 mg hypobaric bupivacaine (4 ml), and the motor and sensory blockade at Th10 was achieved again. The patient reported satisfactory anesthesia each time, and developed no complications. In conclusion, spinal anesthesia can be successful even in cases of severe thoracolumbar kyphoscoliosis. PMID- 16210837 TI - Matrix GLA protein gene polymorphisms: clinical correlates and cardiovascular mortality in chronic kidney disease patients. AB - BACKGROUND: Increased vascular calcification plays an important role in the pathogenesis of cardiovascular events in chronic kidney disease (CKD) patients. It is the result of an active ossification process counteracted by 'protective' proteins, such as matrix GLA protein (MGP). Polymorphisms of MGP have been identified. METHODS: The aim of this study was to define the distribution of two MGP polymorphisms (-7, -138) in 99 hemodialysis (HD) patients, in 26 patients with CKD stage 3 and in 135 age- and sex-matched healthy controls. Patients were followed up for 12 months to record any cardiovascular deaths. The cause of death was determined by medical doctors, considering the medical history of each patient. The primers were designed with Primer Express software. RESULTS: MGP 138TT homozygotes were more frequent in the HD group versus controls (p = 0.0004). Additionally, the frequency of the T allele was significantly higher in the HD group (p = 0.0006). The frequency of the A allele of MGP-7 was significantly higher both in the HD group (p = 0.033) and in the CKD group (p = 0.0017) versus controls. MGP-7 GG homozygotes were significantly less common in the CKD group than in controls (p = 0.037). Combination -138TT -7AA was significantly more frequent in both CKD patients (p = 0.001) and in HD patients (p = 0.029) than in controls. Seventeen out of 99 HD patients experienced fatal cardiovascular events. Sixteen (94.1%) were -138TT homozygotes and either -7AA homozygotes or -7GA heterozygotes. CONCLUSION: This study suggests that CKD and HD patients have a different distribution of MGP gene polymorphism as compared with the normal population. Altered MGP gene polymorphism may be a negative prognostic factor for the progression to end-stage renal disease and for cardiovascular events in CKD patients. PMID- 16210839 TI - Surfactant proteins in innate host defense of the lung. AB - Surfactant proteins A and D (SP-A and SP-D) are members of the collectin family of host defense proteins that are expressed in epithelial cells lining the lung. SP-A and SP-D interact with surfactant lipids in complex ways to determine the structure of tubular myelin, and the ratio of surfactant subfractions that, in turn, influences surfactant uptake and metabolism, respectively. SP-A and SP-D play critical roles in host defense of the lung against diverse viral, fungal, and bacterial pathogens. The collectins bind to the surfaces of microbes via carbohydrate-dependent interaction, aggregating, opsonizing, and enhancing clearance of the organisms by alveolar macrophages in the lung. Pulmonary surfactant proteins A and D play dual roles in pulmonary homeostasis, determining the structure of alveolar lipids and mediating the innate host defense system of the lung. PMID- 16210838 TI - Renal redox stress and remodeling in metabolic syndrome, type 2 diabetes mellitus, and diabetic nephropathy: paying homage to the podocyte. AB - Type 2 diabetes mellitus has reached epidemic proportions and diabetic nephropathy is the leading cause of end-stage renal disease. The metabolic syndrome constitutes a milieu conducive to tissue redox stress. This loss of redox homeostasis contributes to renal remodeling and parallels the concurrent increased vascular redox stress associated with the cardiometabolic syndrome. The multiple metabolic toxicities, redox stress and endothelial dysfunction combine to weave the complicated mosaic fabric of diabetic glomerulosclerosis and diabetic nephropathy. A better understanding may provide both the clinician and researcher tools to unravel this complicated disease process. Cellular remodeling of podocyte foot processes in the Ren-2 transgenic rat model of tissue angiotensin II overexpression (TG(mREN-2)27) and the Zucker diabetic fatty model of type 2 diabetes mellitus have been observed in preliminary studies. Importantly, angiotensin II receptor blockers have been shown to abrogate these ultrastructural changes in the foot processes of the podocyte in preliminary studies. An integrated, global risk reduction, approach in therapy addressing the multiple metabolic abnormalities combined with attempts to reach therapeutic goals at an earlier stage could have a profound effect on the development and progressive nature to end-stage renal disease and ultimately renal replacement therapy. PMID- 16210840 TI - Neonatal chronic lung disease in the post-surfactant era. AB - This is a brief review of neonatal chronic lung disease, sometimes called the 'new bronchopulmonary dysplasia (BPD)'. The clinical, radiographic and pathological features of this condition have changed considerably in recent years because of major advances in perinatal care, including widespread use of antenatal glucocorticoid therapy, postnatal surfactant replacement and improved respiratory and nutritional support. Authentic animal models, featuring lengthy mechanical ventilation of surfactant-treated, premature neonatal baboons and lambs, have provided important insights on the pathophysiology and treatment of this disease. Lung histopathology after 2-4 weeks of positive-pressure ventilation with oxygen-rich gas results in failed formation of alveoli and lung capillaries, excess disordered elastin accumulation, smooth muscle overgrowth in small pulmonary arteries and airways, chronic inflammation and interstitial edema. Treatment interventions that have been tested in these animal models include nasal application of continuous positive airway pressure, high-frequency mechanical ventilation, inhaled nitric oxide and retinol. The challenge now is to improve understanding of the molecular mechanisms that regulate normal lung growth and development, and to clarify the dysregulation of lung structure and function that occurs with injury and subsequent repair so that effective treatment or prevention strategies can be devised and implemented. PMID- 16210841 TI - Patent ductus arteriosus and respiratory outcome in premature infants. AB - A persistent ductus arteriosus is a common event in preterm infants. The systemic to-pulmonary shunting that occurs as the pulmonary vascular resistance decreases after birth can have significant cardiovascular and respiratory consequences. Acute pulmonary effects include pulmonary edema and hemorrhage, worsened lung mechanics and deterioration in gas exchange with hypoxemia and hypercapnia. The increased pulmonary blood flow can also produce damage to the capillary endothelium and trigger an inflammatory cascade. This, plus the need for longer and more aggressive mechanical ventilation, can explain the association between patent ductus arteriosus and an increased risk for bronchopulmonary dysplasia in extremely premature infants. PMID- 16210842 TI - The clever fetus: responding to inflammation to minimize lung injury. AB - Clinical and experimental information indicate that fetal exposure to inflammation can induce lung maturation. This inflammation may be chronic and indolent. We present clinical and experimental information that challenge assumptions about chorioamnionitis associated infection/inflammatory exposures to the fetus. We question the assumption that the fetal compartment is sterile, and that delivery is inevitable if chronic infection/inflammation is present. We demonstrate that the preterm fetus can develop a brisk and adequate inflammatory response, and the fetus also can quickly modulate and downregulate inflammation to prevent injury. The fetus at risk of early preterm delivery may be exposed to both chorioamnionitis and glucocorticoids. Both exposures can either increase or decrease fetal inflammatory responses depending on the timing of the exposures. The immunomodulatory ability of the fetus to fetal exposures remains an unexplored research field. PMID- 16210843 TI - Methylxanthine therapy for apnea of prematurity: evaluation of treatment benefits and risks at age 5 years in the international Caffeine for Apnea of Prematurity (CAP) trial. AB - Methylxanthine therapy reduces the frequency of apnea and the need for mechanical ventilation. Recent research has raised concerns about the safety of methylxanthines in very preterm infants. Possible adverse effects include poor growth, worsening of hypoxic-ischemic brain damage and abnormal childhood behavior. Over 2,000 infants with birth weights 500-1,250 g have been randomized in the international placebo-controlled Caffeine for Apnea of Prematurity (CAP) trial to examine the long-term efficacy and safety of methylxanthine therapy for the management of apnea of prematurity. Additional therapies such as continuous positive airway pressure were used as necessary to control apneic attacks. At 18 months we measure the combined rate of death or survival with one or more of the following impairments: cerebral palsy, cognitive deficit, blindness and deafness. This outcome was chosen because of the need to evaluate the impact of common neonatal therapies beyond discharge from the intensive care unit. However, several potential long-term consequences of methylxanthine therapy may not become apparent until the study cohort reaches pre-school age. We will therefore extend the follow-up to age 5 years. The main outcome at 5 years will be a composite of death or survival with severe disability in at least one of six domains: cognition, neuromotor function, vision, hearing, behavior, and general health. Once this project is completed, caffeine will be one of the most rigorously evaluated neonatal therapies. PMID- 16210844 TI - Dysregulation of expression of immunoregulatory and cytokine genes and its association with the immaturity in neonatal phagocytic and cellular immunity. AB - BACKGROUND: Innate and adaptive immunity is comprised of cellular and humoral factors that provide rapid protection against microbial invasion. However, immaturity of innate and adaptive immune responses in the perinatal period predisposes the neonate to increased infectious morbidity and mortality from a variety of organisms. OBJECTIVES: To elucidate dysregulation of expression of various immunoregulatory and cytokine genes and its association with the immaturity in neonatal phagocytic cellular immunity. METHODS: Comparison of protein production and mRNA of granulocyte macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte colony stimulating factor (G-CSF), interleukin (IL)-12, IL-15 and IL-18 in adult peripheral blood (APB) mononuclear cells (MNC) and cord blood (CB) MNC was studied. Effects of hematopoietic growth factors (HGFs, GM-CSF, M-CSF, G-CSF, IL 11) were studied in vivo in rats as well as randomized controlled studies conducted in neonates. Oligonucleotide microarrays were used to study gene expression patterns of activated CB and APB monocytes and dendritic cells. RESULTS AND CONCLUSIONS: We demonstrated dysregulation of various immunoregulatory and cytokine genes in CB MNC. This dysregulation may in part explain the immaturity of neonatal cell-mediated immunity. There are probably various dysregulated cytokines yet to be discovered. Biological agents such as IL 2, IL-12, IL-11 and/or IL-18 alone or in combination with HGFs should be considered for future studies to identify new approaches to enhance neonatal host defense, and thereby decrease the incidence of neonatal sepsis and the consequent high risk of morbidity and mortality. PMID- 16210845 TI - Oxidative stress in the newborn--a 30-year perspective. AB - In this review the development of the concept 'hypoxia-reoxygenation injury' is outlined. An update of some important factors and mechanisms related to oxidative stress injury in newborn infants is presented, including the metabolism of glutathione, the role of antioxidants, iron and nitric oxide, and how these may influence health and disease in the newborn and contribute to 'oxygen radical disease of the newborn'. New insight into how hyperoxia and hypoxia may induce changes leading to retinopathy of prematurity by vascular endothelial growth factor acting in concert with insulin-like growth factor is briefly summarized. Inflammation and oxidative stress seem to be two sides of the same coin in newborn babies both contributing to injury partly through similar mechanisms. PMID- 16210846 TI - IGF-1 and retinopathy of prematurity in the preterm infant. AB - BACKGROUND: Retinopathy of prematurity (ROP) continues to be a major cause of blindness in children. Although ablation of the retina reduces the incidence of blindness by suppressing the neovascular phase of ROP, the visual outcomes after treatment are often poor. Preventive therapy is required and will likely come from a better understanding of the pathophysiology of the disease. OBJECTIVES: To study the role of insulin-like growth factor 1 (IGF-1) and vascular endothelial growth factor (VEGF) in both the proliferative phase of ROP (phase II) and in the early phase when blood vessels are lost. METHODS: Using both a mouse model of ROP and clinical studies the relationship between IGF-1, VEGF and both vessel loss and vessels proliferation in the retina was studied. RESULTS: IGF-1 is required for maximum VEGF activation of vascular endothelial cell proliferation and survival pathways. IGF-1 levels are deficient after premature birth, setting the stage for retinal vascular loss and ROP. CONCLUSIONS: Restoration of IGF-1 to levels found in utero may help prevent ROP. PMID- 16210847 TI - Protein detection in dried blood by surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS). AB - BACKGROUND: The rapid and reliable identification of biomarkers in the smallest possible amount of blood remains a challenge in biomarker epidemiological research involving preterm newborns. OBJECTIVE: We wanted to explore whether the proteomics approach of 'surface-enhanced laser desorption/ionization-time of flight mass spectrometry' (SELDI-TOF MS) is possible and feasible in whole cord blood previously dried on filter paper. METHODS: Umbilical cord blood from 7 healthy newborns was frozen as serum, whole blood (with or without additives), or dried on filter paper (with or without additives). We used the SELDI-TOF MS technique for protein detection on the ProteinChip arrays: weak cationic exchange array (CM10), hydrophobic array (H50), and strong anion exchange array (Q10). Profiles were compared in terms of peak intensity and number of resolved peaks. RESULTS: Dried neonatal blood, eluted from filter paper, revealed profiles similar to the profiles derived from serum at a protein range of 3-10 kDa. Among additives, heparin led to highest peak intensities for both blood and dried blood. Spectra from heparinized whole blood and heparinized dried blood from the umbilical cord of 8 different healthy newborns on three different types of ProteinChip arrays were very similar. CONCLUSION: We conclude that it is possible and feasible to use SELDI-TOF MS for recovery and detection of whole proteins from dried blood collected on filter paper. The method is easy to perform in large groups of newborns, minimizing the amount of blood needed for biomarker studies. The validity and reproducibility of this method needs to be studied in detail. PMID- 16210848 TI - Dietary Echinacea purpurea during murine pregnancy: effect on maternal hemopoiesis and fetal growth. AB - BACKGROUND: The medicinal benefits of Echinacea sp. plants in several disease conditions, including insect bites, respiratory ailments, and even cancer and AIDS, have been touted for decades. Echinacea sp.-based phytoceuticals are among the top selling herbals in the Western marketplace today. However, evidence is very scant concerning the effects of using Echinacea species herbals during pregnancy. While available data indicates that fetal malformations do not occur during pregnancy in humans consuming this herb, there are no formal studies aimed at assessing the possibility that consuming Echinacea herbals may promote spontaneous abortions, thereby reducing the number of live births upon which to assess the presence or absence of malformations. OBJECTIVES: We undertook a study in which pregnant mice were fed daily Echinacea purpurea from pregnancy onset until gestational days 10, 11, 12, 13, and 14. METHODS: Maternal spleen and bone marrow were taken for enumeration of cells in each of five separate hemopoietic lineages/organ, and fetal status was recorded. RESULTS: The data indicate that the significant, pregnancy-induced elevation in splenic lymphocytes and nucleated erythroid cells was all but eliminated in those females which consumed E. purpurea daily throughout their pregnancy. Moreover, consuming E. purpurea during pregnancy reduced the number of viable fetuses. CONCLUSIONS: The data may be extrapolated to suggest that in humans, abstention from consuming Echinacea products during the early/mid stages of pregnancy, may be prudent. PMID- 16210849 TI - Beta1-adrenergic receptors maintain fetal heart rate and survival. AB - Beta-adrenergic receptor (betaAR) activation has been shown to maintain heart rate during hypoxia and to rescue the fetus from the fetal lethality that occurs in the absence of norepinephrine. This study examines whether the same subtype of betaAR is responsible for survival and heart rate regulation. It also investigates which betaARs are located on the early fetal heart and whether they can be directly activated during hypoxia. Cultured E12.5 mouse fetuses were treated with subtype-specific betaAR antagonists to pharmacologically block betaARs during a hypoxic insult. Hypoxia alone reduced heart rate by 35-40% compared to prehypoxic levels. During hypoxia, heart rate was further reduced by 31% in the presence of a beta(1)AR antagonist, CGP20712A, at 100 nM, but not with a beta2 (ICI118551)- or a beta3 (SR59230A)-specific antagonist at 100 nM. Survival in utero was also mediated by beta1ARs. A beta1 partial agonist, xamoterol, rescued 74% of catecholamine-deficient (tyrosine-hydroxylase-null) pups to birth, a survival rate equivalent to that with a nonspecific betaAR agonist, isoproterenol (87%). Receptor autoradiography showed that beta1ARs were only found on the mouse heart at E12.5, while beta2ARs were localized to the liver and vasculature. To determine if the response to hypoxia was intrinsic to the heart, isolated fetal hearts were incubated under hypoxic conditions in the presence of a betaAR agonist. Heart rate was reduced to 25-30% by hypoxia alone, but was restored to 63% of prehypoxic levels with 100 nM isoproterenol. Restoration was completely prevented if beta1ARs were blocked with CGP20712A at 300 nM, a concentration that blocks beta1ARs, but not beta2- or beta3ARs. Our results demonstrate that beta1ARs are located on the heart of early fetal mice and that beta1AR stimulation maintains fetal heart rate during hypoxia and mediates survival in vivo. PMID- 16210850 TI - Effects of an intratracheally delivered anti-inflammatory protein (rhCC10) on physiological and lung structural indices in a juvenile model of acute lung injury. AB - BACKGROUND: Mechanical ventilation results in acute lung trauma that can stimulate processes that alter lung development. Activation of matrix metalloproteinases (MMPs) and their tissue-produced inhibitors (TIMPs) is initiated by the inflammatory response to mechanical ventilation and are involved in breakdown of the basement membrane and parenchymal modeling. OBJECTIVES: The aim of this study was to test the hypothesis that rhCC10, a lung anti inflammatory mediator, would foster improved lung function, structural preservation, and a reduction in net MMP activity in a juvenile model of acute lung injury. METHODS: Twenty-four juvenile rabbits were saline-lavage-injured and treated with 100 or 25 mg/kg surfactant (Survanta, Ross Labs) with or without rhCC10 (Claragen, Inc.; n=6 per group). Animals were ventilated for 4 h, then euthanized for in vitro surfactant function analysis, lung histomorphometry, and analysis of MMP-2, MMP-7, and MMP-9 and TIMPs 1 and 2 in the lung. RESULTS: Apical lung expansion, reduced with the lower dose of surfactant, was partially restored with the addition of rhCC10. Alveolar septal wall thickness was reduced (p<0.05) with low-dose surfactant plus rhCC10 compared to high-dose surfactant alone. Increased within-group variance in MMP-2 and MMP-9 proteolytic activity was found with the low-dose surfactant and was abolished with rhCC10. MMP-7 was reduced (p<0.05) with rhCC10 administration, independent of surfactant dose. CONCLUSIONS: Intratracheal administration of the anti-inflammatory rhCC10 resulted in preserved lung structure and MMP/TIMP profile after 4 h of mechanical ventilation, in a surfactant dose-dependent manner. PMID- 16210851 TI - Frequencies of A(TA)7TAA, G71R, and G493R mutations of the UGT1A1 gene in the Malaysian population. AB - BACKGROUND: Gilbert syndrome is caused by defects in the uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene. These mutations differ among different populations and many of them have been found to be genetic risk factors for the development of neonatal jaundice. OBJECTIVES: The objective was to determine the frequencies of the following mutations in the UGT1A1 gene: A(TA)7TAA (the most common cause of Gilbert syndrome in Caucasians), G71R (more common in the Japanese and Taiwanese population), and G493R (described in a homozygous Malay woman with Crigler-Najjar syndrome type 2) in a group of Malaysian babies with hyperbilirubinemia and a group of normal controls. METHODS: The GeneScan fragment analysis was used to detect the A(TA)7TAA variant. Mutation screening of both G71R and G493R was performed using denaturing high performance liquid chromatography. RESULTS: Fourteen out of fifty-five neonates with hyperbilirubinemia (25%) carried the A(TA)7TAA mutation (10 heterozygous, 4 homozygous). Seven out of fifty controls (14%) carried this mutation (6 heterozygous, 1 homozygous). The allelic frequencies for hyperbilirubinemia and control patients were 16 and 8%, respectively (p=0.20). Heterozygosity for the G71R mutation was almost equal among both groups (5.5% for hyperbilirubinemia patients and 6.0% for controls; p=0.61). One subject (1.8%) in the hyperbilirubinemia group and none of the controls were heterozygous for the G493R mutation (p=0.476). CONCLUSIONS: The A(TA)7TAA seems more common than the G71R and G493R mutations in the Malaysian population. PMID- 16210852 TI - Ipsilateral ptosis associated with anterior thalamic infarction. PMID- 16210853 TI - Transient neurologic symptoms related to cerebral amyloid angiopathy: usefulness of T2*-weighted imaging. PMID- 16210854 TI - The impact of age and intrauterine contraception on the clinical course of pelvic inflammatory disease. AB - BACKGROUND: The aim of the study was to investigate the clinical course of pelvic inflammatory disease (PID) and factors that could predict failed conservative treatment of PID. Additionally, the study aimed to examine the role of age and intrauterine device (IUD) use on the severity of PID. METHOD: Fifty-one women admitted to hospital with the diagnosis of acute PID were recruited. Of those, 17 patients were subsequently operated because of failed conservative treatment. All women underwent careful examination and completed a questionnaire at admission. Their clinical course was followed and the clinician responsible for the patient completed forms at admission and at discharge. Two groups were established retrospectively, those who were treated conservatively and those who underwent surgery. The outcome results were analyzed with regard to IUD use, duration of IUD use (>or=5 or <5 years), and with regard to age below or above 35 years. All data were analyzed using the statistical package SAS. A p value <0.05 was considered significant. RESULTS: Women who subsequently underwent surgical treatment were significantly older and significantly more frequently, current IUD users. There was no significant difference with regard to other socio-demographic characteristics. Women who subsequently underwent surgery had significantly more frequent complaints of severe abdominal pain, elevated body temperature, symptoms of peritoneal irritation, and appearance of adnexal mass. No differences were found between groups with regard to anaerobic microbiological findings, nor with regard to the finding of Actinomyces. IUD use and age >or=35 were found to be highly significant risk factors for surgery in patients with PID. CONCLUSION: Age over 35 years and IUD use, independently of each other, were factors strongly associated with an increased risk of surgery for PID as a result of failed conservative treatment. PMID- 16210855 TI - Catastrophe theory model for decelerations of fetal heart rate. AB - BACKGROUND: Decelerations of fetal heart rate (FHR) are the results of many pathophysiological factors that modulate the intrinsic rate of the heart. FHR change is a complex phenomenon and can be viewed as a type of nonlinear dynamic system. This paper presents a qualitative model of FHR decelerations based on catastrophe theory, especially to account for abrupt heart rate changes seen in variable decelerations. METHODS: One of the elementary catastrophes, the cusp catastrophe, was used to model FHR changes seen in variable, late, and early decelerations. RESULTS: Catastrophe theory describes typical variable decelerations as an abrupt response to a slowly changing force of attack (uterine contraction) and physiological defense. Because of its three-dimensionality and topological features, the cusp catastrophe model provides a qualitatively satisfying description of FHR decelerations. CONCLUSION: The organized complexity of living systems offers the best hope for the application of catastrophe theory. The nature of the elementary catastrophes will be essential knowledge in the mathematical analysis of biology. PMID- 16210856 TI - Association of angiotensinogen gene polymorphisms with essential hypertension in African-Americans and Caucasians. AB - OBJECTIVE: Molecular variants of angiotensinogen (AGT) have been linked to essential hypertension, and promoter variants have been shown to alter the transcription rate of AGT in vitro. We employed a case-control study to determine whether single nucleotide polymorphisms (SNPs) in the promoter region of AGT were associated with hypertension in African-Americans and Caucasians. METHODS: The frequencies of the variants at base positions -6, -20, -217, -793, and -776, both alone and in combination (haplotypes), were compared between cases and controls in samples stratified based on race and sex. A logistic regression model was applied to test whether AGT genotypes were significant predictors of the disease while adjusting for race, sex, and age. RESULTS: Subjects with the AA or AG genotype at locus -793 were significantly more likely to have the disease (OR = 1.88, 95% CI = 1.12-3.15). Additionally, the differences in haplotype frequency distributions between cases and controls were significant at the 7% level for all four subgroups (stratified by race and sex) after adjusting for multiple testing. Based on the odds ratios for each individual haplotype, the haplotype AAAAT (nucleotide sequences at base positions -6, -20, -217, -793, -776) in African American males, African-American females, and Caucasian females may confer susceptibility to the disease in these population subsets. CONCLUSION: Overall, the present report provides statistical evidence for the association of AGT with essential hypertension. PMID- 16210857 TI - Absent or delayed adrenarche in Pit-1/POU1F1 deficiency. AB - Mutations of the PIT1/POU1F1 gene are responsible for a rare variant of anterior hypopituitarism, including deficiency of growth hormone, prolactin and thyrotropin. In 8 ethnically diverse POU1F1-deficient patients (4 different mutations) with normal circulating levels of cortisol and adrenocorticotropic hormone, and with spontaneous onset and progression of puberty, we observed an absence or delay of adrenarche (median circulating dehydroepiandrosterone-sulfate -6.2 SD); in each of the 4 postmenarcheal females, pubarche (i.e. appearance of pubic hair) was also absent or delayed. The absence/delay of adrenarche in POU1F1 deficient patients and the absence/delay of pubarche in POU1F1-deficient females suggest that a POU1F1-dependent factor contributes to the normal development of adrenarche and female pubarche. PMID- 16210858 TI - Rheumatoid factors: good or bad for you? AB - BACKGROUND: Rheumatoid factors (RFs) are autoantibodies associated with rheumatoid arthritis. They can be detected in normal individuals, although transiently. This dichotomy has led to questions about the origins and types of RFs. Recently it has been shown that B cells that produce RFs only do so when activated by two signals, one from engagement of the B-cell receptor and the other from recognition of a pathogen-associated molecular pattern through a Toll like receptor (TLR). These autoantibodies thus link the innate and acquired immune responses. OBJECTIVE: Through a review of the literature, an examination of the current knowledge of RF induction is presented. The focus is on a discussion of a beneficial or detrimental role for RFs in normal individuals and in those with chronic disease. RESULTS: What makes RF 'good' in some cases and 'bad' in others may reflect the type of RF produced. Low-affinity polyreactive IgM RFs are probably beneficial as they aid in the clearance of immune complexes that are more efficiently cleared, and the RF B cell can act as an antigen presenting cell and stimulate host defense. However, large amounts of high affinity RFs found in patients with chronic disease may be harmful by participation in a vicious cycle of autoantibody production by stimulation of self lymphocytes, and/or deposition in blood vessels thus causing vasculitis. CONCLUSIONS: Whether RFs are beneficial or detrimental depends on the context in which they are expressed, the type and amount of RF produced, whether the response is perpetuated by TLR ligation and whether other cells are stimulated either directly or indirectly by RF-positive B cells. PMID- 16210859 TI - The effect of desloratadine on eosinophil/basophil progenitors and other inflammatory markers in seasonal allergic rhinitis: a placebo-controlled randomized study. AB - BACKGROUND: Eosinophil/basophil (Eo/B) progenitors fluctuate in the peripheral circulation during seasonal allergen exposure in atopic subjects. Several drugs have been shown to modulate Eo/B progenitor levels in the peripheral blood but, to date, the possible effect of antihistamines on Eo/B progenitors has not been explored. Our objective was to evaluate whether the antihistamine desloratadine (DL) can modulate peripheral blood Eo/B progenitors or other markers of allergic inflammation. METHODS: We performed a randomized double-blind placebo-controlled study on the effects of DL on peripheral blood Eo/B progenitors in subjects with symptomatic, seasonal allergic rhinitis during a ragweed pollen season. Forty five subjects were randomized to treatment for 4 weeks with DL 20 mg daily or placebo. RESULTS: The expected fall in the number of Eo/B progenitors from baseline to 2 weeks of treatment was seen in the placebo group [median drop of 1.0 colony-forming unit (CFU)/10(6) cells], and was greater than in the DL group (median drop of 0.0 CFU/10(6) cells) (p = 0.013). The change in histamine concentration per colony from baseline to 2 weeks of treatment was lower in the DL group (median decrease of 6.1 pg/colony) compared to placebo (median increase of 1.8 pg/colony) (p = 0.01). An increase in the nasal lavage eotaxin concentration from baseline to 4 weeks of treatment was statistically significant in the placebo group but not in the DL group. Eo/B CFU were not affected by varying in vitro concentrations of DL. CONCLUSION: These results suggest that DL can modulate aspects of allergic inflammation in vivo through mechanisms other than simple blockade of H1 histamine receptors. PMID- 16210860 TI - Allergic sensitisation in tuberculosis and leprosy patients. AB - BACKGROUND: A negative association has been observed between infections and allergy in several studies. The aim of the present study was to examine whether tuberculosis and leprosy patients have more or fewer allergies than healthy individuals. METHOD: Sera from tuberculosis patients, leprosy patients and healthy controls were analysed by ELISA and Pharmacia Unicap for serological markers for allergy and mycobacterial infection. The serological markers for allergy were total IgE, specific IgE using Phadiatop and specific IgE to the dust mite allergen Dermatophagoides pteronyssinus 1 (Der p 1). Serological markers for mycobacterial infections included specific IgG to a mixture of bacille Calmette Guerin culture filtrate antigens, to purified mannose-capped lipoarabinomannan (manLAM) and to purified secreted antigen 85B. RESULTS: Both tuberculosis and leprosy patients had significantly higher levels of total IgE than controls. Furthermore, a significantly higher level of specific IgE (Phadiatop) was also found in the tuberculosis patients compared with controls. A similar result, but not statistically significant, was observed for the leprosy group. Specific IgG to antigen 85B and to manLAM was found to be significantly higher in both tuberculosis and leprosy patients compared with controls. In addition, leprosy patients had significantly more IgG to the BCG culture filtrate antigen than controls. CONCLUSIONS: The results indicate that patients with mycobacterial infections have allergic sensitisation more frequently compared with healthy controls. This is seemingly in contrast with the notion that there is a negative association between allergy and infection ('hygiene hypothesis'). However, since only one in ten of those infected with Mycobacterium tuberculosis will develop the disease, patients with active mycobacterial disease represent a selected group. A similar relationship applies for leprosy. It is conceivable that those predisposed to allergy are less resistant to mycobacterial infections. PMID- 16210861 TI - Are the asthma guideline goals achieved in daily practice? A population-based study on treatment adequacy and the control of asthma. AB - BACKGROUND: The guidelines for asthma recommend that the use of anti-inflammatory therapy should be adapted to the severity of the disease. However, few data are available to assess the adequacy of the use of drugs and its influence on the control of asthma in 'real life'. METHODS: The adequacy of the current use of anti-asthmatic medication according to the Global Initiative for Asthma (GINA) guidelines was assessed in a random sample of 400 asthmatics identified in the frame of the Italian Study on Asthma in Young Adults. Asthma severity was assessed using the GINA criteria; accordingly, a patient was classified as receiving inadequate treatment if his/her current use of drugs was lower than that suggested by the guidelines for the corresponding severity level. The absence of asthma attacks in the last 3 months was used as an indicator of the disease control. RESULTS: Fifty-five percent of the patients had persistent asthma. Overall, 48% (95% CI 41.2-54.8) of persistent asthmatics were receiving inadequate treatment, and 66% (95% CI 59.5-72.4) had not used their medication daily over the past 3 months. Persistent asthmatics who were inadequately treated had a significantly greater frequency of asthma attacks (geometric mean ratio 3.7; 95% CI 2.1-6.6) than those using an adequate dose of medication. Mild and moderate persistent asthmatics using an adequate medication regimen reported a low number of asthma attacks (median 0). At the multivariate analysis, a good control of the disease was positively associated with an adequate dose of anti inflammatory medication (OR = 2.2; 95% CI 1.1-4.5) and was negatively associated with a later onset of asthma (OR = 0.96; 95% CI 0.93-0.99) and severe asthma (OR = 0.37; 95% CI 0.17-0.81). CONCLUSIONS: Despite the increase in the use of inhaled corticosteroids, half of the persistent asthmatics from the general population are using a medication regimen below their severity level. When the use of drugs follows the GINA guideline recommendations, a good control of asthma is also achievable in the daily management of the disease, particularly in the case of mild and moderate asthmatics. PMID- 16210862 TI - Epidemiological study of muscular disorders in Assiut, Egypt. AB - Few comprehensive epidemiological studies of the prevalence of muscle diseases have been undertaken, and none has been carried out in our locality. The present cross-sectional study was conducted in Assiut Governorate (Upper Egypt) to estimate the prevalence of different types of primary muscular disorder in 1997. The study involved 52,203 subjects, 15,617 (30%) from the rural community and 36,586 (70%) from the urban community. Patients were identified from a door-to door survey, and all were subjected to a full clinical examination, with confirmation of the diagnosis through electrophysiological, and biochemical investigations. Histopathological studies were performed for the classification of muscular dystrophies. Forty patients with muscular disorders were identified, with a point prevalence of 76.6 per 100,000 in the total community with no significant differences between the rural and urban communities. The creatine kinase level was abnormally high (>225 IU/l) in 80% of the cases, increased in all patients with muscular dystrophy or myositis, in 88.8% of patients with systemic myopathy and 66.6% of patients with myotonia. None of the cases of myasthenia showed an increase in the creatine kinase level. The lifetime prevalence per 100,000 was 26.8 for muscular dystrophy, 11.49 for myotonia, 11.49 for myositis, 17.24 for systemic myopathy and 9.57 for myasthenia. PMID- 16210863 TI - Demyelination in the brain as a paraneoplastic disorder: candidates include some cases of multiple sclerosis. PMID- 16210864 TI - Comment on "Inter-rater reliability of delirium rating scales". PMID- 16210866 TI - Naloxone treatment prevents prenatal stress effects on peritoneal macrophage activity in mice offspring. AB - The present study analyzed the effects of maternal stress (PS) and/or naloxone treatment on the activity of peritoneal macrophage in male and female Swiss mice offspring. Pregnant female rats received a daily footshock (0.2 mA) and/or a naloxone injection from gestational day 15 to 19. Experiments were performed on postnatal day 30 on male and female pups. The following results were obtained in male offspring: (1) PS decreased both the index and the percentage of phagocytosis, this decrement being reversed by naloxone treatment, and (2) naloxone alone decreased the percentage of phagocytosis. The following results were obtained in female offspring: (1) PS decreased spontaneous and phorbol myristate acetate-induced macrophage oxidative burst, this decrement being reversed by naloxone pretreatment, and (2) PS decreased both the index and percentage of the phagocytosis, this effect was prevented by naloxone treatment. These data are discussed focussing on a putative neuroimmune interaction involving opioidergic systems during the ontogeny of the central nervous and immune systems. PMID- 16210867 TI - Individual differences in the expression of tyrosine hydroxylase mRNA in neurosecretory neurons of the human paraventricular and supraoptic nuclei: positive correlation with vasopressin mRNA. AB - Previous studies indicated that in the human paraventricular nucleus (PVN) and in the supraoptic nucleus (SON) tyrosine hydroxylase (TH) - the first and rate limiting enzyme in catecholamine synthesis - is localized mainly in magnocellular neurosecretory neurons. Individual differences were observed among control subjects in number and distribution of TH-immunoreactive (IR) perikarya, indicating that antemortem factors may regulate TH expression. Since a large number of TH-IR perikarya were observed in subjects who suffered from somatic illnesses leading to prolonged osmotic or nonosmotic stimulation of vasopressin (VP) release, we suggested that TH expression is related to the activation of VP neurons. The purpose of our study was to apply (1) in situ hybridization for TH mRNA on human PVN and SON to investigate how the previously reported individual differences in TH protein expression are depicted at the transcriptional level and (2) quantitative TH immunohistochemistry and in situ hybridization for VP mRNA throughout the dorsolateral part of the SON (dl-SON) in order to elucidate whether indeed expression of TH in neurosecretory nuclei depends on activation of VP neurons. Postmortem formalin-fixed, paraffin-embedded hypothalamic sections of 16 control subjects were studied for TH protein and TH and VP mRNAs. For 6 of the above cases, the number of TH-IR neurons and the total VP mRNA levels were estimated throughout the entire dl-SON using an image analysis system. Individual variation was observed in TH mRNA expression which appears to parallel the expression of TH-protein. Using Spearman's bivariate test, a positive correlation was found between the number of TH-IR- and TH-mRNA-expressing neurons in both PVN and SON (p < 0.01) as well as between the number of TH-IR neurons and the total VP mRNA in the dl-SON (p < 0.05). Our results show (1) that the individual variability in the number of TH-IR neurons within the neurosecretory nuclei might be due to differential expression and/or stability of TH mRNA and (2) that expression of TH-immunoreactivity in human PVN and SON depends on the activation of VP neurons. PMID- 16210869 TI - Diversity of gastric carcinogenesis. AB - Gastric cancer is still the second leading cause of cancer death worldwide, although the incidence of this disease has been gradually decreasing. The diversity of gastric cancer is well known. However, the mechanism underlying this diversity is still unknown. We have performed experimental and clinical studies on gastric carcinogenesis. These results suggest that the variety in the type and extent of inflammation in the stomach, and the different response to inflammation are responsible for the diversity in gastric carcinogenesis. This concept is supported by clinical and experimental evidence that eradication of Helicobacter pylori diminishes the incidence of gastric cancer and that PPAR gamma ligand, a modulator of inflammation and cell differentiation, prevents gastric carcinogenesis. PMID- 16210868 TI - A novel growth hormone secretagogue-1a receptor antagonist that blocks ghrelin induced growth hormone secretion but induces increased body weight gain. AB - Ghrelin, the natural ligand for the growth hormone secretagogue-1a (GHS-1a) receptor, has received a great deal of attention due to its ability to stimulate weight gain and the hope that an antagonist of the GHS-1a receptor could be a treatment for obesity. We have discovered an analog of full-length human ghrelin, BIM-28163, which fully antagonizes GHS-1a by binding to but not activating the receptor. We further demonstrate that BIM-28163 blocks ghrelin activation of the GHS-1a receptor, and inhibits ghrelin-induced GH secretion in vivo. Unexpectedly, however, BIM-28163 acts as an agonist with regard to stimulating weight gain. These results may suggest the presence of an unknown ghrelin receptor that modulates ghrelin actions on weight gain. In keeping with our results on growth hormone (GH) secretion, BIM-28163 acts as an antagonist of ghrelin-induced Fos protein immunoreactivity (Fos-IR) in the medial arcuate nucleus, an area involved in the ghrelin modulation of GH secretion. However, in the dorsal medial hypothalamus (DMH), a region associated with regulation of food intake, both ghrelin and BIM-28163 act as agonists to upregulate Fos-IR. The observation that ghrelin and BIM-28163 have different efficacies in inducing Fos-IR in the DMH, and that concomitant administration of ghrelin and an excess of BIM-28163 results in the same level of Fos-IR as BIM-28163 administered alone may demonstrate that in the DMH both ghrelin and BIM-28163 act via the same receptor. If so, it is unlikely that this receptor is GHS-1a. Collectively, our findings suggest that the action of ghrelin to stimulate increased weight gain may be mediated by a novel receptor other than GHS-1a, and further imply that GHS-1a may not be the appropriate target for anti-obesity strategies. PMID- 16210870 TI - Stem cell relationships and the origin of gastrointestinal cancer. AB - Gastrointestinal stem cells have the capacity for long-term self-replication and the ability to give rise to all other epithelial cell lineages. These properties make them essential since they maintain tissue homeostasis by regulating cell turnover depending on the current demand. However, they are also important players in the earliest stages of gastric and colonic cancer, as they form a target for mutations to accumulate and lead to the development of the malignant phenotype. Due to the lack of reliable markers, gastrointestinal stem cells are difficult to define and characterise. This limits the knowledge about their number and position within the gastric gland and the intestinal crypt, respectively, and consequently about the clonal structure of these units. Therefore, the morphological events of early gastrointestinal carcinoma formation and expansion are hotly debated. In this review we summarize the properties of gastrointestinal stem cells and illuminate their role in the development of the earliest lesions in the gastric and colonic mucosa. We also resume current opinions about the morphological pathways and the clonality of these neoplasias and the subsequent mechanism of spread within the adjacent tissues. PMID- 16210871 TI - Cancer invasion and metastasis. AB - Metastasis is the transfer of malignant tumors from one organ to a distant organ. It is the most common cause of death in cancer patients. Different molecular mechanisms enable tumor cells to infiltrate the surrounding tissue, invade blood vessels and leave the blood stream at a different site. Tumor cell interaction with extracellular matrix components and epithelial-mesenchymal transition as well as angiogenesis are important factors in invasion and metastasis. Gene expression profiles of metastatic cells in various organs are different and not every tumor cell has the capacity to metastasize. The microenvironment in the organ influences formation of metastasis. Only 1% of micrometastases progress into macrometastases. PMID- 16210872 TI - Genes involved in invasion and metastasis of gastric cancer identified by array based hybridization and serial analysis of gene expression. AB - Gastric cancer (GC) is still a serious health problem and remains the second most common type of fatal cancer worldwide. Comprehensive gene expression analyses may be useful to identify genes associated with invasion/metastasis in GC. Among them, array-based hybridization and serial analysis of gene expression (SAGE) are currently the most common approaches. Over the past 3 years, several large-scale gene expression studies with array-based hybridization and SAGE have been performed and several genes have been identified. This review describes genes associated with invasion/metastasis in GC which have been identified by array based hybridization and SAGE. We compared the expression levels of the genes identified by array-based hybridization with our SAGE data. In addition, expression of the candidate genes obtained by SAGE was further investigated by quantitative RT-PCR of 40 GC samples. MIA and GW112 were overexpressed in 10 (25%) and 22 (55%) of 40 GC samples, and the overexpression of these two genes was associated with tumor stage, respectively. We also discuss the significance of HMGB1/amphoterin in invasion and metastasis of GC. PMID- 16210873 TI - The molecular signature of metastases of human hepatocellular carcinoma. AB - The current metastasis paradigm suggests that the primary tumor starts off benign but over time slowly acquires changes that provide a few rare cells within the tumor the ability to metastasize. However, this concept has been challenged by several recent studies using the microarray-based approach. We have recently found that the molecular signature of primary hepatocellular carcinoma (HCC) is very similar to that of their corresponding metastases, while it differs significantly in primary HCCs with or without metastasis. Similar findings are also evident in primary cancers of the lung, breast, and prostate. Such a signature can be used to predict the prognosis of HCC patients. Moreover, there are significant differences in the gene expression profiles of liver parenchyma among HCC patients with or without intrahepatic metastases. These findings imply that many of the metastasis-promoting genes are embedded in the primary tumors and that the ability to metastasize may be an inherent quality of the tumor from the beginning. In addition, the condition of liver parenchyma may dictate the intrahepatic metastasis potential, which is consistent with the hypothesis that the degree of viral-hepatitis-mediated liver damage or possibly the genetic makeup of individuals may play an important role in metastasis. PMID- 16210874 TI - Mechanisms for the prevention of gastrointestinal cancer: the role of prostaglandin E2. AB - Carcinoma of the colon or rectum represents one of the most common malignancies worldwide with a higher prevalence in industrialized regions. Epidemiologic studies of individuals taking non-steroidal anti-inflammatory drugs (NSAIDs) have shown a significant reduction in colorectal cancer (CRC) mortality compared to those individuals not receiving these agents. NSAIDs inhibit the enzymatic activity of both isoforms of cyclooxygenase (COX-1 and COX-2), while COX-2 selective inhibitors have shown some efficacy in reducing polyp formation. COX-2 derived bioactive lipids, including the primary prostaglandin (PG) generated in colorectal tumors, PGE(2), are known to stimulate cell migration, proliferation and tumor-associated neovascularization while inhibiting cell death. Here we briefly review the role of NSAIDs in preventing CRC, as well as the proposed mechanism by which a COX-2-derived PG, PGE(2), promotes colon cancer. PMID- 16210875 TI - Cyclooxygenase-2 inhibition prevents colorectal cancer: from the bench to the bed side. AB - Cancer is predicted to become the leading cause of death--surpassing heart disease--by the end of this decade. Colorectal cancer is a major health concern, with more than 1,000,000 new cases and 500,000 deaths expected worldwide per year. There is much evidence to suggest a link between the consumption of non steroidal anti-inflammatory drugs (NSAIDs) and the prevention of colorectal cancer (CRC). The consumption of NSAIDs is not problem free, and the number of deaths due to NSAIDs equals the number of deaths from AIDS or leukemia. Therefore, although chemoprevention of CRC is possible, drugs that have more acceptable side effect profiles than the currently available NSAIDs are required. Since up to 50% of polyps and 85% of colonic tumors in humans overexpress cyclooxygenase (COX-2), COX-2 inhibitors are an ideal drug candidate for CRC prevention or treatment. PMID- 16210876 TI - Cancer prevention by phytochemicals. AB - Information has been accumulated indicating that diets rich in vegetables and fruits can reduce the risk of a number of chronic diseases, including cancer, cardiovascular disease, diabetes and age-related macular degeneration. Phytochemicals (various factors in plant foods), such as carotenoids, antioxidative vitamins, phenolic compounds, terpenoids, steroids, indoles and fibers, have been considered responsible for the risk reduction. Among them, a mixture of natural carotenoids has been studied extensively and proven to show beneficial effects on human cancer prevention. PMID- 16210877 TI - Biliary tract malignancy and chronic inflammation from the perspective of pancreaticobiliary maljunction. AB - This is a brief review of pancreaticobiliary maljunction. The basic treatment for this condition includes either cholecystectomy or extrahepatic bile duct resection. When the condition is accompanied by malignancies, a radical operation should be the first treatment option. Knowledge on molecular pathogenesis is gradually increasing. However, studies should be expanded to include larger patient cohorts, and other types of molecules should be carefully investigated and analyzed. PMID- 16210878 TI - Chronological trend of calorie intake and the incidence of epithelial neoplasms of the large intestine during the past 30 years in Japan. AB - OBJECTIVES: The authors tried to elucidate the effects of food intake on the incidences of colorectal cancer and adenoma. MATERIALS AND METHODS: Large intestines obtained from a series of consecutive autopsies performed at the Tokyo Metropolitan Geriatric Hospital were scrutinized to detect colorectal neoplasms. Chronological trends and age-dependent incidences of adenoma and overt lethal cancer were checked, while cancer in adenoma were included in the category of adenoma. Data on food consumption were obtained from the 'The National Nutrition Survey in Japan' (Ministry of Health, Labor and Welfare of Japan). RESULTS: There were 405 cancers and 12,883 adenomas in the 7,091 large intestines examined. The incidences of cancer and adenoma increased with age; overt lethal cancer was found in 5.79% and adenoma in 56.79% of the general population > or =60 years in Tokyo. Calorie intake steeply increased since 1964, the year of the Olympic Games in Tokyo, until the early 1970s. Subsequently, it has continuously declined down to the level of the late 1940s, although the ingredients have changed tremendously. Chronological trends of the incidences of colorectal cancer and adenoma showed similar patterns as calorie intake, but, the influence of calorie intake on the incidences of cancer or adenoma was manifested 18 and 24 years later, respectively. CONCLUSIONS: (1) During the last 30 years, the incidences of colorectal cancer and adenoma were 5.79 and 56.79%, respectively, in individuals aged > or =60 years in Japan. (2) The chronological trends of both lesions showed a pattern similar to that of calorie intake, but, as the trend of cancer incidence preceded the course of adenoma by 6 years, adenoma is not the sole precursor lesion of overt lethal cancer. PMID- 16210879 TI - Neostigmine, but not metoclopramide, abolishes ethosuximide-induced functional gastrointestinal disturbances. AB - Ethosuximide is a drug used for the treatment of absence seizures. Its prolonged application leads to gastrointestinal (GI) disturbances. The purpose of the present study was to determine the severity and nature of the disturbances, as well as the influence exerted upon them by neostigmine methylsulfate and metoclopramide. The drug-induced alterations, occurring in the rat GI tract, were registered by X-ray contrast examination. In vitro experiments were carried out on isolated GI smooth muscle (SM) preparations. The latter revealed that the drug hyperpolarized the SM tissues and inhibited their contractile activity. X-rays of ethosuximide-treated rats showed GI hypotonia, disturbed peristalsis and decreased evacuation activity. The inhibition of the GI functions was associated with hyperpolarization of SM and a reduction in Ca(2+) influx, ensuring spontaneous contractile activity. The application of neostigmine methylsulfate significantly removed ethosuximide-induced functional GI disturbances in rats treated for 15 days with ethosuximide. PMID- 16210880 TI - Influence of epithelium and isoprenaline incubation on responsiveness of guinea pig trachea to methacholine. AB - There are reports regarding harmful effect of long-term use of beta2-agonist drugs on asthma severity and airway responsiveness. In the present study, the responses of guinea pig trachea with intact and denuded epithelium (groups 1 and 2, n = 10) to methacholine as EC50 were measured in tissues nonincubated or incubated with 10 mumol/l isoprenaline during the resting period. The same protocol was performed in groups 3 and 4 (n = 5 for each group) with an additional 30 min rest time after isoprenaline incubation. The response of trachea with denuded epithelium (groups 2 and 4) to methacholine was significantly higher than that with intact epithelium both in incubated and nonincubated conditions (groups 1 and 3, p < 0.05 to p < 0.001). Incubation with isoprenaline caused a significant reduction in the tracheal response to methacholine in both the denuded groups (p < 0.005 and p < 0.001) and intact epithelium groups (p < 0.005 for both cases). The reduction in tracheal responsiveness to methacholine due to incubation in epithelium denuded trachea (groups 2 and 4) was nonsignificantly greater than that of intact epithelium tissues. There was no difference between groups 3 and 4 with those of groups 1 and 2 in both incubated and non incubated conditions. The maximum contractility response to methacholine was not different between tracheal chains with denuded and intact epithelium and did not change due to incubation with isoprenaline. The results of this study indicate reduction of tracheal response to methacholine due to incubation of tissues with isoprenaline, which was relatively more pronounced in epithelium denuded trachea. PMID- 16210881 TI - Immunogenetics of severe respiratory infections: models for the development of new therapeutic strategies. AB - Innate and adaptive immunity plays a critical role in the defence of the lung and other mucosal surfaces exposed to micro-organisms. Anti-microbial peptides and proteins, cytokines and chemokines are important immune weapons as they build up the protective front for the respiratory tract. The notion that susceptibility to infectious diseases may be inherited is widely accepted and, as it is the failure to activate adaptive immunity that may allow infection to become established and progress toward invasion and dissemination, the recognition of specific gene defects affecting the ability of the immune system to overcome invading pathogens may shed light upon those mechanisms of immune regulation that are playing the most critical roles. The aim of the present review is to discuss some of the advances in infection immunogenetics that may lead to identify new strategies in the development of new anti-infectious and anti-inflammatory drugs. PMID- 16210882 TI - Sleep apnea and insomnia: sleep apnea plus or sleep apnea minus. PMID- 16210883 TI - Insomnia subtypes and their relationships to daytime sleepiness in patients with obstructive sleep apnea. AB - BACKGROUND: Insomnia and obstructive sleep apnea syndrome (OSA) are the two most common sleep disorders. Studies showed that insomnia complaints were prevalent in OSA. Relatively little is known about the causes of insomnia in OSA and whether etiological factors differ for insomnia subtypes. We hypothesized that sleep onset problem was mainly due to hyperarousal and sleep maintenance difficulty was primarily related to sleep-disordered breathing. OBJECTIVES: To determine the prevalence of insomnia subtypes in OSA, compare subjects with sleep onset insomnia, sleep maintenance insomnia and no insomnia symptom, and study the relationship of insomnia subtypes to daytime sleepiness. METHODS: We analyzed intake questionnaires and polysomnography and Multiple Sleep Latency Test results of 157 OSA patients. RESULTS: Forty-two percent of the sample had at least one problematic insomnia symptom. The prevalence of sleep onset insomnia, sleep maintenance insomnia and insomnia with early awakening was 6, 26 and 19%, respectively. Patients with sleep onset insomnia had significantly lower apnea hypopnea (AHI) and arousal indices. There were significant inverse relationships between sleep onset insomnia and measures of daytime sleepiness. On the contrary, subjects with repeated wakening had more severe subjective sleepiness. Results were similar in patients with AHI >or=5 or >or=15. CONCLUSIONS: Insomnia symptoms were common in OSA patients. Insomnia subtypes related differently to measures of daytime sleepiness. Our findings suggest that OSA patients with sleep onset insomnia may be in a state of hyperarousal. It is clinically relevant to examine insomnia subtypes, which may influence the treatment decision in sleep-disordered breathing. PMID- 16210884 TI - An acute bronchodilator test with tiotropium or salmeterol does not allow a subdivision of patients according to responses. AB - BACKGROUND: The possibility that individual chronic obstructive pulmonary disease (COPD) patients respond better to either tiotropium or salmeterol has been suggested and an acute bronchodilator test might help to guide the choice of therapy. OBJECTIVES: We explored the responses to tiotropium and salmeterol within the first 3 h using the recommended dosages (18 microg for tiotropium and 50 microg for salmeterol) in order to verify whether there are differences in the short-term bronchodilator effects between these two long-acting bronchodilators in patients with stable COPD. Moreover, we investigated whether these differences could discriminate between the two agents. METHODS: Forty consecutive patients with COPD, but in a stable phase of the disease, participated in this double blind, double-dummy, randomized crossover study. The study involved a screening visit and 2 study days separated by at least 1 week. For determination of the onset of action, we measured FEV1 at 10, 20, 30, 60 and 90 min, and again 2 and 3 h after inhalation of single study drug. RESULTS: At all time points, both tiotropium and salmeterol caused significant (p< 0.001) changes from baseline. A mean increase of 12% above baseline was reached 66 min (95% CI: 45-87) after tiotropium and 59 min (95% CI: 41-77) after salmeterol. Nine patients after tiotropium and 5 patients after salmeterol did not have this type of improvement. A mean increase of 200 ml above baseline was reached 76 min (95% CI: 52-101) after tiotropium and 76 min (95% CI: 52-100) after salmeterol. Eighteen patients after tiotropium and 15 patients after salmeterol did not have this improvement. Twenty-one patients after tiotropium and 24 patients after salmeterol reached an improvement that was at the same time 12% and 200 ml greater than baseline. CONCLUSIONS: These data clearly show that the bronchodilator effects of tiotropium and salmeterol within the first hours after their acute administration are similar in patients with stable COPD when they are evaluated as mean changes from baseline in FEV1. Therefore, they question the direct therapeutic relevance of a subdivision of patients according to bronchodilator responses to tiotropium or salmeterol. PMID- 16210885 TI - Prevalence of chronic obstructive pulmonary disease according to BTS, ERS, GOLD and ATS criteria in relation to doctor's diagnosis, symptoms, age, gender, and smoking habits. AB - BACKGROUND: Guidelines and standards for diagnosis and management of chronic obstructive pulmonary disease (COPD) have been presented by different national and international societies, but the spirometric criteria for COPD differ between guidelines. OBJECTIVES: To estimate prevalence of COPD using the guidelines of the British Thoracic Society (BTS), the European Respiratory Society (ERS), the Global Initiative for Chronic Obstructive Lung Disease (GOLD), and the American Thoracic Society (ATS). Further, to evaluate reported airway symptoms, contacts with health care providers, and physician diagnosis of COPD in relation to the respective criteria, and gender differences. METHOD: In 1992 a postal questionnaire was sent to a random sample of adults aged 20-69 years, 4,851 (85%) out of 5,681 subjects responded. In 1994-1995 a random sample of the responders, 970 subjects, were invited to a structured interview and a lung function test; 666 (69%) participated. RESULTS: The prevalence of COPD was 7.6, 14.0, 14.1, 12.2 and 34.1% according to BTS, ERS, GOLD, clinical ATS (with symptoms or physician diagnosis), and spirometric ATS criteria, respectively. Prevalent COPD was related to age, smoking habits and family history of obstructive airway disease but not to gender. Physician diagnosis of chronic bronchitis or emphysema was only reported by 16.3, 12.2, 11.0, 23.4 and 8.2% of subjects fulfilling the respective criteria, though a majority reported airway symptoms. CONCLUSION: The main determinants for prevalent COPD were age, smoking habits and spirometric criteria of COPD. Though a majority reported airway symptoms and contact with health care providers due to respiratory complaints, only a minority was diagnosed as having COPD, indicating a large underdiagnosis. PMID- 16210886 TI - Once-daily administration of fluticasone propionate does not worsen controlled airway hyperresponsiveness in patients with asthma. AB - BACKGROUND: Inhaled steroids are currently the most important drugs for asthma patients, but compliance tends to be low. Compliance could be improved by reducing the number of daily administrations. OBJECTIVES: In the present study, we compared once- and twice-daily administration of fluticasone propionate (FP) to determine the differences in efficacy. METHODS: Subjects were 40 patients diagnosed with bronchial asthma with stable symptoms and pulmonary functions who were on twice-daily FP administration of 100 microg. There were 14 men and 26 women ranging from 29 to 72 years of age. After a 4-week observation period, subjects were randomized into two administration groups by the envelope method and followed for 8 weeks: group A, once-daily administration (200 microg of FP at night), and group B, twice-daily administration (100 microg of FP in the morning and at night). Clinical symptoms, pulmonary functions and airway responsiveness were compared between these two groups. RESULTS: No significant deterioration in clinical symptoms, pulmonary functions and airway responsiveness were observed in group A compared with group B. CONCLUSIONS: These results demonstrate that once daily FP administration is as effective as twice-daily administration, and that it may improve the compliance for inhaled steroids. PMID- 16210887 TI - C-reactive protein in lymphocytic pleural effusions: a diagnostic aid in tuberculous pleuritis. AB - BACKGROUND: C-reactive protein (CRP) pleural fluid levels have been found to be higher in tuberculosis and parapneumonic effusions than in other causes of pleural effusion. OBJECTIVE: The aim of this study was to analyze whether CRP (a simple and inexpensive test) may be a diagnostic aid for tuberculosis in lymphocytic pleural effusions. METHODS: One hundred and forty-four patients with a lymphocytic pleural effusion (more than 50% lymphocytes in the differential white blood cell count) were included. The patients were 93 men (65%) and 51 women (35%), aged 64 +/- 18 years (mean +/- SD). The diagnoses were as follows: tuberculosis, 20; pleural effusion associated with malignancy, 69; transudates, 38; other benign exudates, 17. RESULTS: The CRP pleural fluid level was higher in tuberculous pleuritis (54 +/- 24 mg/l) than in lymphocytic effusions of other origin (21 +/- 16 mg/l; p < 0.001). High CRP levels (>or=50 mg/l) have a high specificity for tuberculosis (95%), and low levels (<30 mg/l) have a high sensitivity (95%) for excluding disease. CONCLUSIONS: CRP pleural fluid level determination is useful in the diagnostic workup of lymphocytic pleural effusions. High CRP levels are very suggestive of tuberculous pleuritis, and low CRP levels make this diagnosis unlikely. PMID- 16210888 TI - Cell profiles of bronchoalveolar lavage fluid as prognosticators of idiopathic pulmonary fibrosis/usual interstitial pneumonia among Japanese Patients. AB - BACKGROUND: The role of bronchoalveolar lavage fluid (BALF) cell profiles in predicting the clinical outcome of idiopathic pulmonary fibrosis (IPF)/usual interstitial pneumonia (UIP) is still under discussion. OBJECTIVE: To determine whether BALF cell profiles affect the survival of patients with UIP diagnosed by surgical lung biopsy/autopsy at the early stage of IPF. METHODS: This hospital based retrospective cohort study used 81 Japanese patients with histologically proven IPF/UIP who underwent BAL examination. The BALF samples were obtained from non-current smokers: NCS (n = 41) and current smokers: CS (n = 40). The Kaplan Meier and Cox's proportional hazard methods were used to estimate the survival and evaluate the risk ratio for death in the two groups. To detect the multicollinearity, a stepwise regression was employed. RESULTS: A slight increase in the absolute numbers of BALF neutrophils tended to relate to a decrease in the relative risk for death in NCS patients and CS patients in the univariate analysis. In stepwise regression, the increase in percent vital capacity and the increase in the BALF CD4/CD8 ratio in NCS was detected as a favorable predictor, while increased BALF cells affected the results due to chronic smoking in CS. CONCLUSIONS: Based on the study bias of the biopsy-proven IPF/UIP patients at stable stages, an independent variable indicating a favorable outcome was an increased BALF CD4/CD8 ratio in NCS patients, while it was difficult to identify definite prognosticators in CS patients. PMID- 16210890 TI - Cardiopulmonary exercise testing in Fabry disease. AB - BACKGROUND: Fabry disease is a rare X-linked disorder that results from a deficiency in a lysosomal enzyme known as alpha-galactosidase A, with accumulation of globotriaosylceramide (Gl3). Early manifestations include angiokeratomas, acroparesthesias, and hypohidrosis and may progress to renal failure, cardiac dysfunction, and stroke. Patients exhibit decreased exercise tolerance and often complain of fatigue. OBJECTIVE: Our study evaluates the cardiopulmonary characteristics in a cohort of Fabry disease patients at rest and during exercise. METHODS: Thirty-nine patients with a diagnosis of Fabry disease underwent a health screening history and physical examination, an electrocardiogram, an echocardiogram, pulmonary function testing (spirometry), and a non-invasive cardiopulmonary exercise test. A control group was selected for comparison. RESULTS: Eighteen of the 39 Fabry patients (46%) exhibited a significant decrease in diastolic blood pressure (DBP) during exercise. The average decrease in DBP was 10 mm Hg. The maximum drop in DBP was 44 mm Hg. The drop in DBP was evident in 9 of the 24 female patients (38%). None of the control patients had a significant drop in DBP during exercise. CONCLUSIONS: Our finding of a significant decrease in DBP in patients with Fabry disease may explain deficits in exercise tolerance. It is notable that this abnormality is manifested in female patients, even though they are typically not as severely affected as males. PMID- 16210889 TI - The association of adenocarcinoma and hemosputum in pulmonary malignancies. AB - BACKGROUND: Hemosputum was considered one of the popular symptoms of patients with centrally located carcinoma of the lung, especially squamous cell or small cell type. Recent studies reported a relative increase in adenocarcinoma compared with squamous cell carcinoma, especially in developed countries. OBJECTIVES: The main purpose of this study was to determine the trend of histological type associated with the occurrence of hemosputum in patients with pulmonary malignancy. METHODS: Retrospectively, the data from 1,050 patients with pulmonary malignancy who underwent bronchoendoscopic examination at our institution between May 1993 and May 2000 were analyzed. RESULTS: One hundred and forty-nine out of 1,050 patients had hemosputum. There were 66 patients with adenocarcinoma, 55 with squamous cell carcinoma, 15 with small cell carcinoma, 5 with large cell carcinoma, 3 with other cell type carcinoma and 5 with metastatic carcinoma. On bronchoendoscopic examination, abnormal findings in the segmental or more proximal bronchi were found in 82 patients, including 36 with squamous cell carcinoma, 31 with adenocarcinoma, 12 with small cell carcinoma, 2 with large cell carcinoma and 1 with metastatic carcinoma. On the other hand, 67 patients were diagnosed with pulmonary malignancy in the subsegmental or more distal area, including 35 with adenocarcinoma, 19 with squamous cell carcinoma, 3 with small cell carcinoma, 3 with large cell carcinoma, 3 with other cell type carcinoma and 4 with metastatic carcinoma. CONCLUSIONS: The most frequent histological type of malignancy with hemosputum was adenocarcinoma. The number of abnormalities in the peripheral region with hemosputum showed a progressive increase. The number of adenocarcinoma with hemosputum was increased in both central and peripheral regions. PMID- 16210892 TI - Noninvasive bilevel positive pressure ventilation in patients with blunt thoracic trauma. AB - BACKGROUND: Noninvasive bilevel positive pressure ventilation (N-BiPAP) has an established role in providing respiratory support in patients with acute respiratory failure. The significant advantage of N-BiPAP is to avoid endotracheal intubation and its complications. Currently there are no data that support N-BiPAP as first-line treatment in patients with blunt thoracic trauma. OBJECTIVE: To evaluate the safety and efficacy of N-BiPAP in patients with acute respiratory failure due to blunt thoracic trauma. METHODS: Prospective observational study. Twenty-two patients with blunt chest trauma (mean injury severity score 26 +/- 9) were studied. N-BiPAP was applied via a tight-fitting full or total-face mask, combined with regional anesthesia in all patients. RESULTS: N-BiPAP resulted in significant changes in blood gasses, heart rate and breathing frequency at 1 h. Eighteen out of 22 patients avoided intubation and were discharged from the ICU (success group). Four patients met predefined criteria and required intubation (failure group) within 24 h after N-BiPAP. Three of the patients in the failure group survived while 1 developed septic shock and died. The acute response of oxygenation to N-BiPAP differed significantly between groups, being higher in the success group. Complications related to N-BiPAP were minor, consisting of nose bridge injury (1 patient) and gastric distention (1 patient). CONCLUSIONS: N-BiPAP administration could be a safe and effective method to improve the gas exchange in patients with acute respiratory failure due to blunt thoracic trauma. PMID- 16210891 TI - The effect of red cell transfusion on nitric oxide diffusing capacity. AB - BACKGROUND: The diffusion capacity of the lung for nitric oxide (DLNO) is supposed to reflect the properties of the alveolocapillary membrane better than the diffusion capacity of the lung for carbon monoxide (DLCO), due to a much stronger binding of NO to haemoglobin (Hb). OBJECTIVES: The aim of this study was to investigate the effect of Hb concentration on the DLNO. METHODS: The DLNO and DLCO (single-breath method) were measured in 10 anaemic patients before and shortly after red cell transfusion. RESULTS: The mean increase in Hb concentration was 2.6 g/dl. Whereas DLCO increased as predicted by the reference equations, the DLNO did not change: mean DLCO rose from 13.6 to 16.5 ml/min/mm Hg (increase of 122%), mean DLCO corrected for Hb rose from 18.8 to 19.3 ml/min/mm Hg (103%) and mean DLNO rose from 75.6 to 77.8 ml/min/mm Hg (103%). CONCLUSION: The DLNO is not influenced by Hb concentration. PMID- 16210893 TI - Quantitative assessment and repeatability of chlorine in exhaled breath condensate. AB - BACKGROUND: Airway condition is presumably reflected in epithelial lining fluid (ELF). Exhaled breath condensate (EBC) has been used as a surrogate marker of the composition of ELF. OBJECTIVES: This study aimed at assessing the technical repeatability of chlorine measurements in EBC and comparing two separate condensators (Ecoscreen and R Tube regarding recovery and repeatability. Furthermore, the association between condensate recoveries and variations in the airway status were scrutinized. METHODS: EBC was collected using two condensators from 10 healthy volunteers. In addition, 13 asthmatic patients produced EBC with or without an added resistance of 5 cm H2O (Res5), applied to the outflow tract of Ecoscreen. All tests were done in random order. Chlorine levels (analyzed by a coulometric technique) in EBC served as a tool for investigation. RESULTS: Chlorine was measurable in all samples. The coefficient of repeatability of chlorine measurements was <10%. Chlorine levels were higher in EBC obtained from R Tube (p < 0.001), and differences in recoveries and variability in chlorine levels were presumably related to technical differences in the condensators and not to the repeatability of chlorine measurements per se. Air-flow-dependent chlorine levels were obtained from healthy volunteers. Application of Res5, recruiting additional alveoli, resulted in increased recovery of the EBC volume, but not of chlorine, from those that had the most pronounced airway obstruction (p = 0.05). CONCLUSION: We conclude that by employing a sensitive analysis technique, chlorine is repeatedly measurable in EBC. We suggest that the bulk of chlorine in EBC originates from large airways and not from the alveolar area. Both condensators were comparable regarding repeatability but differed regarding chlorine recovery. PMID- 16210894 TI - Evaluation of tracheal imaging by optical coherence tomography. AB - BACKGROUND: Optical coherence tomography (OCT) is a new technology capable of generating high resolution cross-sectional images of complex tissue in real time. Analogous to ultrasound, OCT measures backscattered light intensity using coherence interferometery to construct topographical images of complex tissue. Since OCT uses infrared light rather than acoustic waves, its spatial resolution is exceptionally high (2-10 microm). Recent advances in data acquisition, analysis, and processing enable real-time imaging, and make OCT a potentially valuable tool for pulmonary airway diagnostic applications, including assisting directed airway biopsies. OBJECTIVE: This study evaluates feasibility of OCT for delineating proximal airway microstructures in various animal as well as human tracheas. METHODS: Excised trachea samples from New Zealand white rabbits, Duroc pigs, and human trachea were imaged using a compact, 1,300-nm broad-band superluminescent-diode-based prototype fiber OCT device we constructed. The resulting structural OCT images were compared to conventional hematoxilin and eosin (HE) stained histological sections from the same samples. RESULTS: OCT was able to delineate microstructures such as the epithelium, mucosa, cartilage, and glands in all samples. CONCLUSION: These findings suggest that integration of OCT with flexible fiberoptic bronchoscopy could enhance pulmonary diagnostic medicine and detection of pathologic tissue changes in various respiratory diseases. PMID- 16210895 TI - Secondary echinococcosis of the rib and soft tissues. PMID- 16210896 TI - An unusual endobronchial foreign body: a gauze that migrated from the mediastinum. AB - The authors report a 48-year-old white male with lung cancer who was admitted to their clinic with a 1-year history of cough and shortness of breath. The patient was hospitalized because of progression of his complaints over 1 month. On the 3rd day of hospitalization, he expectorated a piece of gauze. The authors performed a fiber-optic bronchoscopy and saw pieces of gauze originating from granulation tissue located in the angle of the lower trachea and left main bronchi. A detailed anamnesis revealed that a mediastinoscopy had been performed 3 years ago for lung cancer staging: the gauze had migrated from the mediastinum to the endobrochial space. PMID- 16210897 TI - Chylothorax following innominate vein thrombosis: a rare complication of transvenous pacemaker implantation. AB - A case of chylothorax following innominate vein thrombosis, which developed as a late complication of transvenous pacemaker implantation, is discussed. A 78-year old man presented with a refractory left-sided pleural effusion, which turned out to be chylothorax. He had undergone a transvenous pacemaker implantation 6 years earlier for sick sinus syndrome. The aetiological workup showed occlusion of the innominate vein as the cause for the chylothorax. The chylothorax resolved following pleurodesis with talc slurry, and the innominate vein was recanalized by angioplasty. To our knowledge this is the first report of a case of this nature. PMID- 16210898 TI - The lung, an organ for absorption? AB - This review summarizes information concerning the mechanisms of absorption of substances across the pulmonary epithelium. Inhalation is now increasingly used as a route of administration, although the scientific understanding of these mechanisms is rather limited. The aim of this study is to draw attention to these questions. PMID- 16210899 TI - Helminth-induced immunoregulation of an allergic response to food. AB - Work from our laboratory has shown that an enteric helminth infection can act as an adjuvant to prime for a Th2-biased response to a typically tolerogenic form of dietary antigen. Helminth infection did not, however, prime for an allergic response. Using a model in which systemic anaphylactic symptoms and antigen specific IgE are induced in C3H/HeJ mice by repeated intragastric administration of peanut antigen with the mucosal adjuvant cholera toxin we showed that an enteric helminth infection protects against the development of food allergy. Helminth-dependent protection against allergy was abrogated when the helminth infected, allergen-sensitized mice were treated with neutralizing antibodies to IL-10. Recent work from our laboratory and others has implicated helminth induced immunoregulatory cells in protection against allergy. We will discuss the characteristics of the immunoregulatory cell populations that have been described and the mechanism(s) by which they may function in the suppression of allergy. PMID- 16210900 TI - The mutual influence of nematode infection and allergy. AB - Several studies have now shown that the prevalence of helminth infections is negatively correlated with the prevalence and/or severity of allergic diseases. Here, we describe studies in rodents infected with Strongyloides venezuelensis examining the mutual influence of nematode infection and allergy. S. venezuelensis has a lung cycle, much akin to the human hookworm and Strongyloidiasis, and induces airway eosinophilia, local IgE and mucus production, and airway hyperreactivity. Both the Th2 and functional responses are relevant for the ability of rodents to deal with S. venezuelensis infection. Nevertheless, the parasite elicits the release of cytokines, such as IL-10, which are capable of regulating immune and functional manifestations. In infected animals, allergic inflammation prevents parasite migration and establishment. Nevertheless, the parasite is capable of regulating the allergic response, preventing part of the tissue damage and functional changes induced by allergy. Understanding the mechanisms by which helminths regulate inflammation may potentially lead to the development of strategies aimed at controlling unwanted inflammation in allergic and autoimmune diseases. PMID- 16210901 TI - Human schistosomiasis decreases immune responses to allergens and clinical manifestations of asthma. AB - Studies have demonstrated that people living in areas endemic for helminths have a decreased reactivity to skin prick tests to aeroallergens and milder forms of asthma. Hypotheses to explain the inverse correlation between helminth infections and atopy include competition between helminth-induced polyclonal IgE and aeroallergen-specific IgE for high-affinity receptors present on mast cells, increased number of regulatory T cells, and high levels of regulatory cytokines, such as IL-10, produced during helminthic infections. Indeed, cells from asthmatic individuals infected with Schistosoma mansoni produce lower levels of IL-5 than asthmatics free of infections. In contrast, IL-10 is more readily produced by allergen-stimulated cells from asthmatics who are infected and is detected only at low levels by cells from helminth-free asthmatics. It is well known that Th2 cytokines are involved in the pathogenesis of allergies and asthma, and some studies indicate that IL-10 is the key cytokine that inhibits the Th2-inflammatory response in allergy. In this chapter we will discuss the association between S. mansoni infection, atopy and severity of asthma and possible mechanisms by which individuals living in helminth endemic areas are protected against the development of allergies. PMID- 16210903 TI - Mechanisms underlying helminth- induced Th2 polarization: default, negative or positive pathways? AB - Since the initial description of Th1 and Th2 subsets in the 1980s, there has been enormous progress in identifying the molecular events and the transcriptional factors that regulate Th differentiation in response to a specific stimulus (e.g. antigen dose, co-receptors, cytokines). Although TCR cross-linking and engagement of co-stimulatory molecules are necessary for activation of CD4+ lymphocytes, these two events do not appear in themselves to explain Th1/Th2 commitment. Among pathogens, helminths are the main examples of Th2-cell inducers in both humans and experimental models. This review will focus on our recent findings on the requirements for Th2 polarization by the parasitic trematode Schistosoma mansoni. In particular, we will address the ongoing controversy as to whether Th2 development depends on positive vs. negative vs. the absence of signals from antigen-presenting cells. In addition, we will discuss the similarities between the pathways involved in parasite- and allergen-induced Th2 differentiation. PMID- 16210902 TI - Proteases in helminth- and allergen- induced inflammatory responses. AB - Proteolytic activity is a central biochemical property that endows molecules with intrinsic allergenicity. Thus, the cysteine protease of dust mite, Der p1, the aspartic protease of cockroach, Bla g 2, the serine protease of Aspergillus fumigatus and the bacterial subtilisins are all major allergenic molecules responsible for the increase in asthma and atopic conditions worldwide. These proteases induce Th2-driven inflammatory responses in the airways by disrupting the epithelial cell junctions so that these, and other molecules, gain access to, and alter the function of, underlying cells of the innate immune system (dendritic cells, mast cells, basophils and macrophages) and B and T cells. Helminth parasites secrete proteases to gain entry into their hosts, and to feed on and migrate through tissues. Their action leads to tissue damage and the activation of inflammatory responses dominated by elevated IgE, eosinophilia and Th2 cells, much like allergenic responses. In certain situations, such as in acute infections (especially with zoonotic helminths), proteases secreted by helminths may sensitise individuals to allergens. However, the anti-inflammatory responses observed in chronic helminthiases, involving IL-10 and TGFBeta, that are primarily responsible for controlling immune-mediated damage to the host that is initiated by secreted proteases, coincidentally protects against similar inflammatory damage by allergens. PMID- 16210904 TI - Regulation of dendritic cell function by pathogen-derived molecules plays a key role in dictating the outcome of the adaptive immune response. AB - There is increasing awareness that dendritic cells (DCs) can interpret pathogen inherent signals and play a pivotal role in polarizing Th cell differentiation. Polarized Th1 responses are induced by DCs, which respond to pathogen-derived TLR ligands to mature and produce IL-12 and related cytokines that are instrumental in Th1 cell outgrowth. In contrast, DCs exposed to SEA (soluble egg Ag from the helminth parasite Schistosoma mansoni) retain a (modified) immature phenotype and induce Th2 responses. In addition to providing positive signals for Th1 cell development, DCs activated to mature by TLR-engagement also provide a potent negative signal that prevents the development of Th2 cells. Production of this signal is dependent upon a MyD88-dependent signaling pathway in DCs. In contrast, exposure of DCs to SEA severely limits their ability to respond to inflammatory TLR ligands such as LPS and CpG. Thus as part of their pathogen-specific response programs, DC can exert negative as well as positive signals for Th response polarization. These effects may have powerful and systemic effects on disease outcome. PMID- 16210905 TI - Glycans modulate immune responses in helminth infections and allergy. AB - Infections of humans and animals by parasitic helminths share key features with atopic diseases, such as allergic asthma. Both diseases lead to the induction of high levels of Th2- type cytokines associated with abundant IgE production and eosinophilia. This immunological association has raised strong interest in the nature of the molecules that promote Th2 and regulatory T cell responses, and the molecular mechanism. Complex carbohydrates are potent inducers of Th2 responses, and carbohydrate antigens (Ags) can stimulate the production of different classes of glycan-specific antibodies (Abs), including Th2 associated IgG but also non specific IgE. In this review we focus on the immunological responses towards glycan Ags derived from allergens and parasitic helminths, especially schistosomes. Biological effects of carbohydrate Ags are dependent on recognition of these Ags by carbohydrate- binding proteins (lectins). Cell-surface C-type lectin receptors (CLRs), such as DCSIGN, L-SIGN, the mannose receptor, macrophage galactose binding lectin, and other lectins, such as the soluble collectins and galectin-3, recognize particular glycan Ags of schistosomes and allergens, which may contribute to orchestrate Th2 associated adaptive responses. Remarkably, schistosomes express 'self glycan' Ags that are recognized by CLRs on DCs, whose principal function is thought to capture self-glycan Ags and generate regulatory T-cells to induce tolerance to these Ags. By expressing such self-glycan Ags, schistosomes may deceive the host immune system to their own benefit. The host protects itself against too much damage by down-regulating helminth-induced Th2 immune responses, and may thus simultaneously be protected against excessive Th2 cell-mediated allergic responses. PMID- 16210906 TI - Role of the natural killer T lymphocytes in Th2 responses during allergic asthma and helminth parasitic diseases. AB - The recent discovery that T cells recognize endogenous and foreign lipid and glycolipid molecules presented by CD1 proteins has brought a major contribution in the understanding of innate and adaptive immune response to certain harmless antigens and infectious pathogens. Among (glyco)lipid-reactive T cells, CD1d restricted natural killer (NK) T cells represent a population of innate/memory lymphocytes that, upon stimulation, rapidly release important amounts of immunoregulatory cytokines that in turn can shape the acquired immune response in a Th1 or a Th2 direction. Here we review the general features of these cells as well as their diverse influence in various disease models. A particular emphasis will be placed on the role of NK T cells in the promotion of asthma, a typical Th2-related inflammatory disease. Moreover, recent studies suggest that NK T cells could also be important in the modulation of the host immune response during helminthic infections, generally associated with dominated Th2 responses. Our current understanding of the role and of the mode of NK T cell activation during the initial immunological events that lead to the promotion of Th2 responses will be discussed. PMID- 16210907 TI - The mast cell and gut nematodes: damage and defence. AB - Gut nematode infection induces a dominant type 2 immune response, crypt hyperplasia and mucosal mastocytosis. Despite their strong association with nematode infection, the role of mast cells in the mechanism of worm expulsion is yet to be fully defined. Recent work suggests that they contribute to resistance, aiding the effector mechanisms which ultimately result in worm expulsion. Although it is widely accepted that both connective and mucosal mast cells arise from a common progenitor, it is clear that mucosal mastocytosis is dependent on the presence of type 2 cytokines such as interleukin 4 (IL-4), IL-9, IL-10 and IL 13. Importantly, it is now evident that mucosal mast cells can amplify this protective response, as well as contributing to intestinal pathology. Here we discuss current areas of interest in this field, including the potentially conflicting role that mast cells play in intestinal inflammation. We also highlight the significance of these responses to current ideas relating to parasite infection and allergy. PMID- 16210908 TI - Basophils, basophilia and helminth infections. AB - A growing body of evidence suggests basophils are important components of the human immune response to helminth infections. Basophil numbers are increased in several animal models of helminth infection, and basophils have been shown to release both histamine and IL-4 in response to helminths. Helminth infections typically provoke type 2 immune responses characterized by eosinophilia, elevated levels of Ag-specific and polyclonal IgE, and T cell production of type 2 cytokines such as IL-4, IL-5, and IL-13. IL-4 plays a central role in this type 2 response. As basophils are the only peripheral blood mononuclear cells with the ability to release IL-4 rapidly in response to appropriate stimuli, releasing large quantities of preformed IL-4 within minutes of surface IgE cross-linking, it appears likely that basophils play an important role in amplifying ongoing type 2 immune responses to helminth infections once Ag-specific IgE is present. Basophils may also function to initiate type 2 responses upon first exposure to helminths and to potentially re-establish these responses upon re-exposure. This article reviews basic basophil biology and physiology, evaluates the evidence for the presence of basophilia in helminth infections, and then focuses on the possible roles basophils serve in the immune response to helminth infections. PMID- 16210909 TI - Innate, adaptive and regulatory responses in schistosomiasis: relationship to allergy. AB - Helminth infections have profound effects on the immune system. Here, recent insights in the molecular interactions between schistosomes and the host are described with respect to adaptive but also with respect to innate immune responses. Furthermore, the different mechanisms of immune hyporesponsiveness are depicted with emphasis on regulatory T cells. Finally, the relationship between downregulatory responses and allergy is discussed. PMID- 16210910 TI - Regulatory T cells induced by parasites and the modulation of allergic responses. AB - The inverse relationship between helminth infection and overt allergic reactivity has intrigued medical scientists for three decades. In the past 5 years, detailed epidemiological studies coupled with new experimental model approaches, have substantiated the negative effects of infection on allergic disease manifestation, and begun to provide mechanistic explanations for this fascinating interaction. Several key conclusions can now be drawn. First, the modulation of allergies, such as Th2-dependent pathologies, is not primarily through immune deviation (e.g. switching responsiveness to Th1), as helminth infections themselves drive strong Th2 responses. Second, helminth-infected hosts show similar levels of immune sensitisation to allergens as do uninfected counterparts, but the expression of overt allergic reactivity is suppressed. Third, the down-modulation of allergy in both human and experimental infections is consistent with the activity of T regulatory (Treg) cell populations, which suppress effector mechanisms of both Th1 and Th2 type. PMID- 16210911 TI - RNA interference remarkably suppresses bcl-2 gene expression in cancer cells in vitro and in vivo. AB - Bcl-2 is an anti-apoptotic protein. If the level of Bcl-2 protein can be reduced sufficiently in tumors using RNA interference (RNAi) to target the gene message, the apoptosis of tumor cells may be promoted. In this study, we synthesized 19 nucleotides (nts) small interference RNA (siRNA) constructs suppressing bcl-2 gene expression in human tumor cells (HeLaB2 and BGC-823 cell lines) in vitro. The bcl-2 gene expression levels were significantly reduced when these siRNA were transfected into experimental two tumor cells for 72 hours. The apoptosis process was also examined in the tumor cells. Here we synthesized siRNA from a DNA template under the control of the RNA polymerase III promoter in transfected tumor cells. Using this DNA vector-based approach, we found that the siRNA efficiently and specifically inhibited the synthesis of protein encoded by the bcl-2 gene in HeLaB2 and BGC-823 tumor cells. Tumor growth was inhibited by 66.5% with 2mg/kg pSilencer 3.1H1-bcl-2 in mouse liver tumor-bearing BALB/c mice. This approach may prove to be a valuable clinical technique for the analysis of specific gene functions and gene therapy of malignant tumors that utilize the bcl 2 gene via RNA interference. PMID- 16210912 TI - Nuclear chromatin patterns in 3 glucocorticoid-resistant RPMI 8226 human myeloma cell sub-lines: correlations with cell growth and immunological phenotype. AB - Nuclear morphological alterations associated with glucocorticoid resistance in human myeloma were evaluated by image cytometry in three human myeloma RPMI 8226 cell sub-lines. Resistance was induced by drug selection using prednisone (8226p), methylprednisolone (8226m) and dexamethasone (8226d), respectively. All these three cell sub lines displayed significant glucocorticoid-resistance without cross-resistance to doxorubicin. Nuclear geometry and texture were analyzed on G0/G1-selected cell nuclei and data compared with cell growth characteristics and membrane expression of CD23, CD38, CD44 and CD58 antigens. When compared to the parental RPMI 8226 cell line, glucocorticoid-resistant cells display a progressive chromatin condensation with heterogeneously distributed large chromatin clumps, a phenomenon not observed in the multidrug-resistant CEM VLB cells. These alterations were correlated to the resistance index against glucocorticoids and to the expressions of CD38, and of CD44 variant forms CD44v5 and CD44v7-8 antigens. These data suggest that glucocorticoid resistance in RPMI 8226 cells could be associated with sub-visual specific higher-order chromatin organization changes. Furthermore, these alterations are correlated to the expression of membrane markers associated with tumors aggressiveness. PMID- 16210913 TI - Analysis of the mononuclear inflammatory cell infiltrate in the non-tumorigenic, pre-tumorigenic and tumorigenic keratinocytic hyperproliferative lesions of the skin. AB - BACKGROUND: The keratinocytic hyperproliferative lesions include non-tumorigenic, pre-tumorigenic (actinic keratoses, AK), and tumorigenic (squamous cell carcinomas, SCC) conditions. Although mononuclear inflammatory cell infiltrate (MICs) is a constant feature in these lesions, their immunophenotypic characterization is still incomplete. We hypothesized that the development of non tumorigenic, pre-tumorigenic, and tumorigenic keratinocytic hyperproliferative lesions is associated with alterations in the mononuclear inflammatory cell infiltrate in response to altered antigenicity of the lesional cells. This study tries to test this hypothesis and to characterize MICs in these lesions. METHODS: Fifty lesions (non-tumorigenic lesions, 29; AK, 9 and SCC, 12) were examined using immunoperoxidase staining methods and antibodies targeting histiocytes (CD68), T cells (CD3), B cells (CD20), and T cells with cytotoxic potential (TIA 1). RESULTS: As compared to the normal skin, the development of the keratinocytic hyperproliferative lesions (normal skin; non-tumorigenic; AK and SCC) was associated with a statistically significant increase (p = <0.05) in: (1) CD20+ B lymphocytes (0.0 +/- 0.0 vs. 3.1 +/- 0.5 vs. 7.5 +/- 0.3 vs. 14.5 +/- 5.5); (2) CD68 histiocytes (4.0 +/- 1.0 vs. 26.5 +/- 3.9 vs. 23 +/- 1.9 vs. 41.3 +/- 6.8); (3) CD3+ T lymphocytes (3.0 +/- 1.1 vs. 58.3 +/- 10.3 vs. 54.5 +/- 0.2 vs. 41.0 +/- 16.0); and (4) TIA-1+ cytotoxic T cells (1.8 +/- 0.4 vs. 2.9 +/- 0.7 vs. 9.6 +/- 1.1 vs. 13.7 +/- 5.2). CONCLUSIONS: The increase in the number of infiltrating mononuclear cells in all pathologic lesions compared to normal skin may reflect increased antigenicity of the lesional cells. Both humoral and cell mediated immunity are involved in these lesions. PMID- 16210914 TI - Enhanced therapeutic effect by combination of tumor-targeting Salmonella and endostatin in murine melanoma model. AB - The growth of tumor is angiogenesis-dependent and it often contains hypoxia and necrotic areas. Salmonella VNP20009 could target and replicate in hypoxia and necrotic areas within tumor and induce antitumor effect. Angiogenesis inhibitor endostatin could reduce tumor angiogenesis and inhibit its growth. However, in the phase I trials of VNP20009 and endostatin at the maximum-tolerated dose, no antitumor effects for bacteria therapy and minor therapeutic effects for endostatin treatment were seen. The ineffectiveness of these agents in clinical trials suggests that the combination of these agents with synergic modalities might be necessary. Here we described antitumor effects mediated by the combination of VNP20009 with recombinant human endostatin in B16F10 murine melanoma model with the aim to exploit tumor-targeting of bacteria and anti angiogenesis strategy to enhance therapeutic efficacy. Combination therapy of these agents significantly enhanced antitumor effects by inducing greater tumor growth inhibition, more severe tumor tissue necrosis as well as less blood vessel density than those induced by either of treatments. The findings suggest that the combination of tumor-targeting bacteria with angiogenesis inhibitor might be of value for the treatment of solid tumors. PMID- 16210915 TI - The Cables gene on chromosome 18Q regulates colon cancer progression in vivo. AB - Early events involved in the pathogenesis of colorectal cancer include mutations in the Adenomatous Polyposis Coli tumor-suppressor gene and oncogenic KRAS mutations. Later events include deletions on chromosome 18q, which are observed in a high proportion of colorectal cancers. However, the important tumor suppressor genes targeted by these deletions have not been fully defined. A previous study found Cables is located on human chromosome 18q11-12. Loss of Cables expression as determined by immunohistochemical staining (IHC) occurred in 60-70% of sporadic colorectal cancers that were usually correlated to loss of heterozygosity at 18q. To determine if Cables is an important target for the chromosome 18q deletions, the susceptibility of Cables-/- mice to develop colon tumors was studied. A well characterized colonic carcinogen, 1,2 dimethylhydrazine (DMH) was used as a tumor initiator. Cables-/- mice (n = 25) and the Cables+/+ littermates (n = 25) were treated with subcutaneous DMH injections over 20 weeks to initiate tumorigenesis. The median survival after DMH injections was significantly shorter for the Cables-/- mice compared to Cables+/+ littermates. The total number of colorectal tumors that developed in the Cables-/ mice was 46 tumors versus 21 tumors. The increased numbers of colorectal tumors, as well as shorter survival of the Cables-/- mice provides compelling evidence that Cables could play an important role in the pathogenesis and progression of colon cancer in mice. These data coupled with previous observations support the hypothesis that Cables is a relevant target of the chromosome 18q deletions frequently seen in human colorectal cancer. PMID- 16210917 TI - Potentiation of the antitumoral activity of gemcitabine and paclitaxel in combination on human breast cancer cells. AB - The purpose of this study was to evaluate the antitumoral activity of different gemcitabine-based combination on an experimental model of human breast cancer, in order to identify the most effective treatment and to provide a rationale for clinical investigations. To this end, CG5 breast cancer cells were treated in vitro with gemcitabine followed by epirubicin, doxorubicin, docetaxel or paclitaxel. The reversed sequence was also investigated. Results, analyzed by multiple drug effect/combination index (CI) isobologram, demonstrated that the combination gemcitabine/paclitaxel was the most active showing synergism with a CI of about 0.5 in the two sequences employed. Moreover, the synergistic interaction of gemcitabine and paclitaxel was correlated to a block of the cells in the G0/G1 compartment of cell cycle and to an increase of apoptotic cells compared to each drug. Based on these evidences, the antitumoral efficacy of gemcitabine/paclitaxel combination has been studied in vivo. Mice bearing CG5 human breast xenografts treated with paclitaxel and gemcitabine in combination showed a significant higher inhibition of tumor growth (approximately 70%) compared to that with either agent alone (25%). In conclusion, this study suggests that paclitaxel is the most promising agent for combination protocols with gemcitabine and supports the use of gemcitabine/paclitaxel combination in the clinical management of advanced breast cancer. PMID- 16210916 TI - Identification of OATP1B3 as a high-affinity hepatocellular transporter of paclitaxel. AB - Interindividual variability in paclitaxel and docetaxel pharmacokinetics, toxicity and response is extensive, and largely unexplained. We hypothesized that this is due to affinity of taxanes for an uptake transporter that indirectly regulates elimination pathways. Here, we studied accumulation of [3H]docetaxel and [3H]paclitaxel in Xenopus laevis oocytes injected with cRNA of the liver specific organic anion transporting polypeptide (OATP) family members OATP1B1 (OATP2) or OATP1B3 (OATP8). Taxane transport by OATP1B1 expressing oocytes was not significantly different from that by water-injected controls, whereas uptake by OATP1B3 was 2.2-fold higher for docetaxel (p = 0.0007) and 3.3-fold higher for paclitaxel (p < 0.0001). OATP1B3-mediated paclitaxel transport was saturable (Michaelis-Menten constant, 6.79 microM), time-dependent, and highly sensitive to chemical inhibition. Paclitaxel uptake was not inhibited by ketoconazole or tariquidar. However, uptake was inhibited by the formulation excipient Cremophor (74.4% inhibition, p < 0.0001), cyclosporin A (25.2%, p = 0.005), glycyrrhizic acid (24.6%, p = 0.012), and hyperforin (28.4%, p = 0.003). Consistent with this finding, Cremophor was found to significantly affect the hepatic uptake of paclitaxel in mice. These data suggest that OATP1B3 is a key regulator of hepatic uptake, and may therefore play a role in the variable response to treatment with taxanes. PMID- 16210919 TI - Fine needle biopsy cytology of breast lesions: a review of interpretative difficulties. AB - Screening mammography and greater community awareness of breast carcinoma have led to an increase in fine needle biopsies of the breast. As a consequence, a wide variety of cytologic patterns have been encountered and studied in benign, proliferative, and malignant breast lesions. We review the main reasons for diagnostic difficulties in breast cytology, the situations in which either a false positive or a false negative diagnosis is possible, as well as conditions whose incorrect typing can lead to inappropriate management. PMID- 16210918 TI - Fragmentation and rapid shortening of telomere in HeLa cells in the early phase of hydroxyl radical-induced apoptosis. AB - In order to maintain genomic stability, cells recognize and respond to DNA damage by either cell cycle arrest or apoptosis. Telomeres are the special DNA sequences at chromosome termini that plays an important role in maintaining chromosomal stability. Telomeric DNA has been shown to be much more susceptible to damage than non-telomere sequences. Recent studies suggest that telomere disruption can trigger apoptosis in certain cell types, mimicking a major cellular response to DNA damage. In this work, we studied the DNA damage at the telomere region during hydroxyl radical-induced apoptosis in HeLa cells. It was found that HeLa cells experienced telomere strand fragmentation and rapid telomere shortening following treatment with H(2)O(2) well before caspase-3 activation and apoptosis. The percentage of cells undergoing apoptosis correlated with the extent of telomere strand fragmentation. Introducing telomere oligonucleotide into cells induced cell cycle arrest in S-phase and apoptosis. We speculate that telomere fragments released from chromosomes might serve as a triggering signal in DNA damage induced apoptosis in the HeLa cells. PMID- 16210920 TI - Lymphoplasmacytic lymphoma/waldenstrom macroglobulinemia: an evolving concept. AB - The concept of Waldenstrom macroglobulinemia has evolved from the original description of a clinical syndrome to its more recent designation as a distinct clinicopathologic entity, that is, lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM), in the World Health Organization (WHO) classification and by the participants of consensus meetings on WM. The diagnosis of LPL/WM, however, remains a challenge in daily practice. Distinguishing LPL/WM from other B-cell lymphomas, especially marginal zone B-cell lymphomas, which share overlapping morphologic features, is difficult. The traditional practice of separating LPL/WM from other lymphomas by an arbitrary level of serum IgM is no longer considered valid. The characteristic immunophenotype described for LPL/WM by the WHO classification, that is, CD5(-)CD10(-)CD23-, is observed in 60-80% of neoplasms, but variations from this pattern of antigen expression are common, with CD23 being detected in up to 40% of cases. Lack of a distinct molecular genetic hallmark complicates the distinction of LPL/WM from other B-cell lymphomas. Although the t(9;14) is stated to be present in 50% of cases in the WHO classification, translocations involving the Ig heavy chain including the t(9;14) are actually rare in LPL/WM. Deletion of 6q21-q23, a nonspecific finding, is the most common aberration reported in 40-70% of patients. At the molecular level, the neoplastic clone in most cases has undergone Ig variable gene mutation, but not isotype switching, and the clone retains the capability of plasmacytic differentiation. Currently, the diagnosis of LPL/WM can only be established by incorporating clinical and pathologic findings and excluding alternative diagnoses. In some cases, in our opinion, distinguishing LPL/WM from marginal zone B-cell lymphomas seems arbitrary using currently recommended criteria. PMID- 16210921 TI - Prognostic and predictive molecular markers in DCIS: a review. AB - Eighteen percent of all new breast cancers detected on screening mammography are ductal carcinoma in situ (DCIS), a preinvasive lesion that is highly curable. However, some women with DCIS will develop life-threatening invasive breast cancer. Because the determinants of invasive recurrence are unknown, all women with DCIS require the same treatment (usually with surgery and radiation). Therefore, there is a need to identify biologic markers and create a profile that will provide prognostic information that is more accurate than the currently used van Nuys Index to predict invasive recurrence. In the present review, we examined the many biologic markers studied in breast cancer, describe their main biologic role and their expression in DCIS, and review the various studies regarding their ability to serve as prognostic factors in breast cancer with an emphasis on predicting invasive recurrence in patients with DCIS. This review covers established markers, namely, ER, PR and HER2/neu, that are used routinely to make treatment decisions as well as investigative biologic factors involved in cell proliferation, cell cycle regulation, extracellular molecules, factors involved in extracellular matrix degradation, and angiogenesis. However, controversies exist regarding the value of these prognostic factors, their interrelationship, and their advantages over morphologic evaluation. PMID- 16210922 TI - Current thoughts on the role of the androgen receptor and prostate cancer progression. AB - Androgens play a central role in the development and maintenance of prostate tissue. Treatment of prostate cancer by androgen ablation either surgically or biochemically results in massive cell death and tumor regression. However, this is often followed by the onset of aggressive disease, which is fatal. Various studies have been conducted to understand the mechanism leading to the establishment of aggressive disease following treatment. An interesting comprehensive study recently conducted by Chen et al shows the increase in androgen receptor (AR) transcript to be the key factor in disease recurrence following treatment. This up-regulation in the AR levels is shown to increase sensitivity to low levels of androgen, leading to ligand-dependent downstream gene expression and tumor recurrence. A "mass action" model has been proposed to explain this phenomenon. Moreover, the increase in mRNA levels has been shown to facilitate conversion of classic antagonists to agonists of hormones action by the recruitment of a subset of co-activators to the promoters of AR-responsive genes. This study underscores the importance of ARs in the establishment of prostate cancer and offers several insights into the mechanism by which tumors recur following androgen ablation. The study also prompts several questions about the reason behind the observed up-regulation and also the mechanism by which classic antagonists are rendered agonistic. The need for the design of novel therapeutic analogues is also emphasized. PMID- 16210923 TI - Clinical value of epidermal growth factor receptor expression in primary breast cancer. AB - EGFR expression in primary breast cancer has been extensively investigated for its prognostic and predictive value. However overall there is no consensus on its potential to guide such prognostication. This is largely because of the great heterogeneity in study designs and methods used to assay the EGFR protein. The impetus to standardize such studies is much needed as there are now several tyrosine kinase inhibitors directed against the EGF receptor and phase II trials are showing significant promise. PMID- 16210926 TI - Whither cardiovascular risk prediction? PMID- 16210924 TI - The significance of benign endometrial cells in cervicovaginal smears. AB - The success of the routine Papanicolaou (pap) smear in reducing the incidence and mortality of cervical cancer has been chronicled extensively. Unfortunately, endometrial cancer, the most common malignancy of the gynecologic tract, continues to lack a screening modality of comparable efficacy. It is generally accepted that the Papanicolau test has a low sensitivity for detecting endometrial disease. Nonetheless, it remains true that endometrial cells are not uncommonly identified on routine cervicovaginal smears and along with each case comes an associated responsibility for pathologists to assess cytologic features, assign a potential clinical significance, and make a decision on reporting this finding. When endometrial cells with an entirely normal cytologic appearance are identified on an otherwise unremarkable cervicovaginal smear, the central question raised is whether such cells are exfoliating physiologically or whether their exfoliation is pathologic in response to an underlying endometrial disease. Additionally, in the former scenario, could subsets of patients be defined in which the reporting of this finding is deemed unnecessary in the cytologic report? In this contribution, we explore the clinical significance of finding normal endometrial cells in cervicovaginal smears based on a review of the medical literature of the last half-century. The historical and evidentiary basis for the Bethesda 2001 recommendations, which calls for the reporting of cytologically benign endometrial cells only in patients 40 years and older, is reviewed in detail. PMID- 16210927 TI - HeartScore: a new web-based approach to European cardiovascular disease risk management. PMID- 16210928 TI - Estimating the risk for atherothrombosis: are current approaches sufficient? AB - Atherothrombotic disease is now the leading cause of mortality in the world and there are only a few methods available to estimate risk of vascular disease. Vascular disease risk assessment methods have been developed from a variety of population based studies and the most frequently used approaches include algorithms developed by Framingham (USA), Munster (Germany), and SCORE (Europe). The field of risk estimation is dynamic and inclusion of newer factors and populations is under active consideration and methods such as discrimination and calibration are now available to evaluate the use of risk assessment tools across different world regions. PMID- 16210929 TI - Predictive accuracy of the SCORE risk function for cardiovascular disease in clinical practice: a prospective evaluation of 44 649 Austrian men and women. AB - BACKGROUND: In 2003, a new risk function for cardiovascular risk in clinical practice was developed by the SCORE project group. The aim of this paper was to evaluate the predictive accuracy of the SCORE in a large Austrian population. DESIGN: A prospective, multicentre, longitudinal linkage project. METHODS: Using the 'SCORE risk function for low-risk regions', we calculated the risk of death from cardiovascular and coronary heart disease events over a 10-year period for 44 649 participants aged 40-65 years in the Vorarlberg Health Monitoring and Promotion Programme (VHM&PP). The predicted risks were compared with the 95% confidence intervals (CI) of the observed events. RESULTS: We observed a total of 487 deaths (1.1%; 95% CI 1.0-1.2) for all cardiovascular disease within 10 years, 371 (1.8%; 95% CI 1.6-2.0) in men and 116 (0.5%; 95% CI 0.4-0.6) in women. The SCORE function overestimated cardiovascular mortality and predicted 666 (1.5%) events, 444 (2.2%) in men and 222 (0.9%) in women. Receiver operating characteristics analyses revealed area under the curve values of 0.76 (95% CI 0.74-0.79) for men and 0.78 (95% CI 0.74-0.82) for women. Multivariable analyses showed that obesity (in men only) increased levels of glucose, gamma-glutamyl transferase, triglycerides (in women only), and blue-collar job status (in women only) significantly contributed to the SCORE as additional independent risk factors. CONCLUSION: Although the SCORE over-predicted the mortality pattern in the cohort as a whole, its predictive ability at the individual level still demonstrates a potentially widespread utility in clinical practice. PMID- 16210930 TI - A comparison of Framingham and SCORE-based cardiovascular risk estimates in participants of the German National Health Interview and Examination Survey 1998. AB - BACKGROUND: Overestimation of risk by Framingham risk functions not only in southern but also in northern European populations including Germany, has led to the development of the SCORE risk estimation model. DESIGN: Data of the German National Health Interview and Examination Survey 1998 was used to determine whether SCORE leads to lower estimates of the 10-year absolute risk of fatal cardiovascular disease and fatal coronary heart disease than a Framingham model. Predicted numbers of events were compared with approximations based on national mortality statistics. METHODS: Inclusion criteria followed the recommendations for the use of SCORE: age 30 to 69 years, no previous history of cardiovascular disease and no markedly raised levels of single risk factors (leaving 1811 men and 1955 women for analysis). RESULTS: The SCORE model for high-risk regions (SCORE-HIGH, which is recommended for Germany pending calibration with national data) predicted the highest number of events, followed by the estimations with mortality statistics, the Framingham model and SCORE-LOW (87 fatal cardiovascular disease events versus 77, 62 and 47; fatal coronary heart disease events 62 versus 46, 46 and 30). Agreement on high-risk status, defined as the 10-year risk of fatal cardiovascular disease of 5% or higher now or if extrapolated to age 60, was moderate for both men and women (kappa 0.52 and 0.42 for Framingham and SCORE HIGH). CONCLUSIONS: Our results suggest that SCORE-HIGH may overestimate absolute risk of fatal coronary heart disease and cardiovascular disease in Germany and may need calibration. Furthermore, the limitations of current risk prediction tools emphasize the ongoing need for comprehensive, high-quality and timely European cohort data. PMID- 16210931 TI - The validity of the Finnish Diabetes Risk Score for the prediction of the incidence of coronary heart disease and stroke, and total mortality. AB - BACKGROUND: Cardiovascular disease shares several risk factors with type 2 diabetes. We tested whether the Finnish Diabetes Risk Score (FINDRISC), recently developed in a Finnish population to estimate the future risk of diabetes, would also identify individuals at high risk of coronary heart disease (CHD) and stroke, and total mortality in this same population. DESIGN: Independent risk factor surveys were conducted in 1987, 1992, and 1997 in Finland, comprising 8268 men and 9457 women aged 25-64 years and free of CHD and stroke at baseline. During the follow-up until the end of 2001, 699 incident acute CHD events, 324 acute stroke events, and 765 deaths occurred. METHODS: The data were analysed by using receiver operating characteristic (ROC) curves and the Cox-regression model. RESULTS: The areas under the ROC curves (AUC) were 71% for CHD, 73% for stroke, and 68% for total mortality in men and 78, 68, and 72% in women, respectively. The addition of systolic and diastolic blood pressures, total and high-density lipoprotein cholesterol, and smoking increased the AUC values modestly (the change of the absolute values from 2.6 to 6.5%), but the additional use of plasma glucose had only a slight effect on the AUC values for CHD and stroke. CONCLUSIONS: The FINDRISC is a reasonably good predictor of CHD, stroke and total mortality. PMID- 16210932 TI - Building prediction models for coronary heart disease by synthesizing multiple longitudinal research findings. AB - BACKGROUND: No methodology is currently available to allow the combining of individual risk factor information derived from different longitudinal studies for a chronic disease in a multivariate fashion. This paper introduces such a methodology, named Synthesis Analysis, which is essentially a multivariate meta analytic technique. DESIGN: The construction and validation of statistical models using available data sets. METHODS AND RESULTS: Two analyses are presented. (1) With the same data, Synthesis Analysis produced a similar prediction model to the conventional regression approach when using the same risk variables. Synthesis Analysis produced better prediction models when additional risk variables were added. (2) A four-variable empirical logistic model for death from coronary heart disease was developed with data from the Framingham Heart Study. A synthesized prediction model with five new variables added to this empirical model was developed using Synthesis Analysis and literature information. This model was then compared with the four-variable empirical model using the first National Health and Nutrition Examination Survey (NHANES I) Epidemiologic Follow-up Study data set. The synthesized model had significantly improved predictive power (chi = 43.8, P<0.00001). CONCLUSIONS: Synthesis Analysis provides a new means of developing complex disease predictive models from the medical literature. PMID- 16210934 TI - Predictors of smoking cessation in patients admitted for acute coronary heart disease. AB - BACKGROUND: Smoking cessation is probably the most important single action after a coronary event. In order to increase the effectiveness of smoking cessation programs, it is important to have knowledge of the predictors of smoking cessation. Further, it is unknown whether smoking cessation programs have impact on these predictors. METHODS: Data were obtained from a randomized controlled trial of smoking cessation intervention in 240 smokers aged less than 76 years admitted for myocardial infarction, unstable angina, or cardiac bypass surgery. Baseline characteristics were prospectively recorded. Smoking cessation was determined by self report and biochemical verification at 12 months follow-up. RESULTS: In multivariate logistic regression analysis, a high level of nicotine addiction, low level of self-confidence in quitting and having previous coronary heart disease were significant negative predictors of smoking cessation at 12 months follow-up. Having previous coronary heart disease and a diagnosis other than acute myocardial infarction as a reason for admission were important negative predictors of abstinence in the usual care group, in contrast to the intervention group, although this did not reach a level of significance in the subgroup interaction analyses. A high level of nicotine addiction was a strong negative predictor in both groups. CONCLUSION: A high level of nicotine addiction is an important negative predictor of smoking cessation, even within an individualized smoking cessation program. Smoking cessation intervention seems to be especially effective in patients with previous coronary heart disease and in patients with unstable angina or coronary artery bypass surgery, compared to usual care. PMID- 16210933 TI - Glucose abnormalities and heart failure predict poor prognosis in the population based Reykjavik Study. AB - BACKGROUND: The risk of cardiovascular disease increases progressively with increasing blood glucose from levels well below the diabetic threshold. In the Reykjavik Study the relationship between heart failure and abnormal glucose regulation was already apparent at the level of impaired glucose tolerance. The aim of this study was to determine the prognosis of participants with any glucose abnormality and heart failure and to test whether the combination of these conditions may adversely affect the subsequent prognosis. DESIGN: A prospective population-based study. METHODS: Data from the first visit of 19 381 participants were used. Participants were divided into groups according to their glycaemic and heart failure level, and comparisons were made between the groups and disease free participants serving as a reference group. The risk of mortality and morbidity was calculated with adjustments for main cardiovascular risk factors and ischaemic heart disease. RESULTS: Participants in the reference group were younger, had lower body mass indices and more seldom a history of myocardial infarction compared with diseased groups. Mortality was lowest in the reference group (P<0.0001) increasing to a maximum in participants with the combination of glucose abnormality and heart failure. Prognostically, the mortality risk associated with abnormal glucose regulation was increased but was lower than the risk of diabetes. The risk of a new myocardial infarction was highest in participants with diabetes [hazard ratio (HR) 1.6; 95% confidence interval (CI) 1.3-2.0] or diabetes in combination with heart failure (HR 1.8; CI 1.1-2.7). CONCLUSIONS: Heart failure or glucose abnormalities are related to increased morbidity and mortality. The combination of glucose abnormality and heart failure did, however, not add further to the unfavourable prognosis in the presence of ischaemic heart disease. PMID- 16210935 TI - Fifteen-year risk of major coronary events predicted by Holter ST-monitoring in asymptomatic middle-aged men. AB - BACKGROUND: Ambulatory electrocardiogram monitoring (Holter) with ST-analysis as a measure of myocardial ischemia has in populations with coronary heart disease been shown to predict major coronary events: death, myocardial infarction or coronary revascularization. There has, however, been conflicting evidence regarding the usefulness of this technique in identification of healthy subjects with increased risk for coronary heart disease. The aim of this study was to assess if Holter monitoring with ST-analysis could be used to predict future major coronary events in asymptomatic middle-aged men with a defined aggregation of traditional risk factors for coronary heart disease. METHODS: One hundred and fifty-five asymptomatic participants from the city of Malmo, Sweden, with known levels of conventional cardiovascular risk factors underwent Holter monitoring for analysis of transient ST-segment depression at the age of 55 years. Fifteen years after the Holter monitoring, hospital records, diagnosis and death registries were revisited for major coronary events. RESULTS: An ST-segment depression of 1 mm or greater (0.1 mV) was considered significant for myocardial ischemia and was found in 54 of the 155 men. There were no significant differences in risk factors in the two groups at baseline. The 15-year incidence of a first major coronary event was significantly higher in men with ST-segment depression (39%) than in men without ST-segment depression (20%) (P<0.015). A Holter electrocardiogram could predict future major coronary events with a positive and negative predictive value of 35 and 80%, respectively. CONCLUSIONS: Holter monitoring can be used as a complement to conventional risk factor evaluation in deciding whether or not to treat risk factors for CHD in asymptomatic subjects. PMID- 16210936 TI - A comparison of the associations between risk factors and cardiovascular disease in Asia and Australasia. AB - BACKGROUND: Cardiovascular disease is already the leading cause of death in many Asian populations. Relationships between vascular risk factors and cardiovascular disease may differ in Asian and western populations. Previously, a lack of prospective data has prevented the reliable quantification of such differences, which, if they were shown to exist, would suggest that novel cardiovascular prevention and treatment strategies are required for Asia. DESIGN: An individual participant data meta-analysis of 32 studies from the Asia-Pacific region involving 331 100 subjects (75% from Asia; 25% from the predominantly Caucasian populations of Australia and New Zealand). METHODS: Outcomes were death from coronary heart disease, ischaemic and haemorrhagic stroke. Hazard ratios were estimated from Cox models for systolic blood pressure (SBP), total cholesterol, triglycerides, body mass index, diabetes and current cigarette smoking, stratified by study and sex and adjusted for age, the other risk factors and regression dilution. RESULTS: After an average period of follow-up of 4 years there were 2082 deaths from coronary heart disease, 600 from haemorrhagic stroke and 420 from ischaemic stroke. The direction and strength of the associations between risk factors and cardiovascular outcomes were similar in the two regions, although in two cases there were significant differences. Triglycerides were more strongly associated with coronary heart disease in Australia and New Zealand (P = 0.03), whereas SBP showed a stronger relationship with haemorrhagic stroke in Asia (P = 0.04). CONCLUSIONS: Classical vascular risk factors act similarly in Asian and Caucasian populations; prevention and treatment strategies should thus be similar. Blood pressure reduction should be particularly effective in Asia. PMID- 16210938 TI - Physical performance and physical activity in grown-up congenital heart disease. AB - AIM: To review exercise performance and exercise habits in patients with congenital heart disease (CHD). BACKGROUND AND METHODS: Physical exercise and physical activity has shown beneficial effects on the physical, psychological and social level in adult patients with cardiovascular disease. Favourable effects have also been documented in children with CHD. Exercise testing is preferentially performed on a treadmill in children, with the measurement of gas exchange. RESULTS: An overview of the literature showed that formal exercise testing has frequently documented reduced or suboptimal values for aerobic exercise performance in children with left-to-right shunts (atrial septal defect, ventricular septal defect), valvular heart disease and obstructive anomalies (aortic stenosis, pulmonary stenosis, coarctation of the aorta). Subnormal values for exercise tolerance have also been observed in patients with successfully repaired cyanotic heart disease (tetraology of Fallot, transposition of the great arteries, Fontan operation). An important contributing factor to the impaired exercise performance is the hypoactive lifestyle, as often observed in patients with CHD. This frequently results from parental or environmental overprotection. CONCLUSION: These patients should be stimulated to be physically active, unless medical restriction is imposed. Fortunately, this represents only a small fraction of the total number of congenital heart defects for which sports participation is allowed. PMID- 16210937 TI - Arterial compliance in young children: the role of aerobic fitness. AB - BACKGROUND: Reduced arterial compliance is reflective of vascular dysfunction, which promotes the atherosclerotic process, and is therefore an important predictor of vascular disease. In adults, obesity, age, aerobic fitness, oestrogens and race influence arterial compliance. Although stature and blood pressure are known to influence compliance in children, other determinants are less established. This investigation sought to determine the predictors of arterial compliance in children, assess the extent to which aerobic fitness is related to compliance, and compare compliance between girls and boys. METHODS: Participants (99 children aged 9-11 years, 55 boys) were assessed for aerobic fitness, physical activity level, blood pressure, body mass, percentage fat mass, height, maturity and arterial compliance (large and small). Predictors of compliance were determined using stepwise regression. Second, children were divided into quartiles according to fitness, and arterial compliance was compared using analysis of covariance (ANCOVA). Finally, differences in compliance between girls and boys were assessed using ANCOVA. RESULTS: We found that fitness, blood pressure and height accounted for 37% of the variance in large artery compliance. Mass, fitness, maturity and blood pressure accounted for 44% of the variance in small artery compliance. Children in the highest fitness quartile had greater compliance than children in the two lowest quartiles, by as much as 34%. There were no differences in compliance between girls and boys after adjusting for covariates. DISCUSSION: These data show that aerobic fitness is associated with arterial compliance in 9-11-year-old children, supporting the concept that physical fitness may exert a protective effect on the cardiovascular system. PMID- 16210939 TI - Electrical stimulation of unloaded muscles causes cardiovascular exercise by increasing oxygen demand. AB - BACKGROUND: The development of new strategies to encourage increased levels of physical activity can help to reduce the incidence of cardiovascular disease. A new system of electrical muscle stimulation (EMS) has been developed that attempts to cause an increase in energy expenditure by mimicking the action of shivering in the body. The purpose of this study was to show that this form of EMS is capable of eliciting a cardiovascular exercise response in healthy adults. DESIGN: An observational study. METHODS: Ten healthy volunteers completed a maximal treadmill test and four EMS sessions using a hand-held EMS device that delivered current to the body via five silicone rubber electrodes on each leg. At each session subjects completed 3 min stimulation at each of four stimulation outputs (10, 20, 30 and 40% of maximum output) while cardiopulmonary gas exchange and heart rate (HR) were measured. Physiological responses at increasing levels of stimulation were evaluated. RESULTS: Average (+/-SD) HR and oxygen consumption (VO2) levels of 67+/-11 bpm and 4.7+/-1.2 ml/kg per min at rest, respectively, were increased to 186+/-10 bpm and 44.9+/-9.8 ml/kg per min at peak exercise intensity on treadmill testing. The electrical stimulation was generally well tolerated by the subjects. Subjects demonstrated statistically significant increases in all physiological variables measured with successive increases in stimulation intensity. Peak HR and VO2 at 40% stimulation intensity were 101+/-12 bpm and 14.9+/-4.3 ml/kg per min, respectively. CONCLUSIONS: These results demonstrate that this form of EMS is capable of producing a physiological response consistent with cardiovascular exercise at mild to moderate intensities. It achieves this without producing gross movement of the limbs or loading of the joints. This EMS-induced cardiovascular exercise response could be used to promote increased levels of physical activity in populations unable to participate in voluntary exercise. PMID- 16210941 TI - Career mapping: chart a course for your future. PMID- 16210942 TI - Acceptance as a management tool. AB - Build a healthier work environment by accepting staff members as they are while working with them to change their behavior. PMID- 16210943 TI - Reclaim lost time with better organization. AB - Better desktop organization and time management can improve a new manager's workflow. PMID- 16210944 TI - Positive collaboration integral to EHR use. AB - Merge the separate needs of nurses and physicians to preserve the processes of this relationship within the electronic healthcare record. PMID- 16210945 TI - Medication management: a patient safety priority. AB - The Joint Commission promotes safe and effective medication management through continuous compliance with its medication management standards. PMID- 16210946 TI - Consistent charge nurses improve teamwork. AB - Team perception on a medical-surgical department improved after the implementation of a dedicated charge nurse staffing strategy. PMID- 16210947 TI - Best-practice protocols: preventing central line infection. AB - Compliance with venous access best practices at Johns Hopkins Hospital hinged on nursing and physician management, and quality improvement programs. PMID- 16210948 TI - Drill down with root cause analysis. AB - Review the structure of an effective critical event analysis and suggestions for completing documentation and maximizing knowledge while protecting your organization from litigation. PMID- 16210950 TI - Healthier hospitals? AB - Review AACN's healthy work environments initiative and learn how the standards can be used to build cultures of safety. PMID- 16210951 TI - Men at work. AB - To address rising healthcare demands, nurse managers must identify strategies to recruit and retain greater numbers of men in nursing. PMID- 16210954 TI - Tips for top challenges: understand healthcare's true cost. PMID- 16210955 TI - Genitourinary malignancies in solid organ transplant recipients. AB - Malignancy is a recognized complication of transplantation. Genitourinary cancers are the second most common tumors in transplant recipients with prostate cancer and renal cell carcinoma the most common. Unlike the more common skin malignancies, genitourinary tumors have a significant impact on both graft and patient survival. Surgical and radiation treatments need to consider the location of heterotopic transplants and administration of chemotherapy may need alteration in light of immunosuppression being used. The major genitourinary malignancies and their management will be reviewed in this article with emphasis on the concerns that arise in a transplant recipient. PMID- 16210956 TI - Function of the vascular endothelial growth factor receptors Flt-1 and Flk-1/KDR in the alloimmune response in vivo. AB - BACKGROUND: We have recently reported that vascular endothelial growth factor (VEGF) functions as a proinflammatory cytokine to regulate the trafficking of leukocytes into allografts in the early posttransplant period. VEGF binds two major VEGF receptors: VEGFR-1 (flt-1) and VEGFR-2 (flk-1/KDR). Here, we wished to investigate the expression and function of VEGF receptors in the process of acute allograft rejection in vivo. METHODS: We performed fully MHC-mismatched C57BL/6 (H-2b) into BALB/c (H-2d) vascularized heterotopic murine cardiac transplants and we examined the expression of VEGF and VEGF receptors by immunohistochemistry during acute allograft rejection. Next, we treated mice with specific neutralizing monoclonal antibodies against murine VEGFR-1 and VEGFR-2 and examined their effect on the development of acute allograft rejection by histology and by analysis of graft survival. The intragraft expression of cytokines and chemokines were also evaluated by quantitative real-time PCR analysis. RESULTS: The expression of VEGF, VEGFR-1 and VEGFR-2 were significantly up-regulated during allograft rejection as compared to isografts. Administration of either anti-VEGFR-1 or anti-VEGFR-2 alone failed to inhibit allograft rejection. However, coadministration of both antibodies together inhibited leukocyte infiltration of allografts and prolonged allograft survival. Furthermore, the effect of VEGFR blockade was associated with the downregulation of intragraft cytokine and chemokine expression. CONCLUSIONS: Our data suggest that VEGF-VEGFR interactions function in the alloimmune response in vivo. Targeting VEGFRs may represent a novel therapy to protect allografts following clinical transplantation. PMID- 16210957 TI - Quantitative assessment of collagenase blends for human islet isolation. AB - BACKGROUND: The variability in collagenase blends has been speculated as the single most important determinant of the success or failure in isolated islet yields in clinical islet transplantation. Examination of the formulation and potency of the widely used Liberase HI enzyme blend will uncover possible sources of imprecision. METHODS: High performance liquid chromatography (HPLC) and kinetic measurements of collagenase and protease activity were used to assess potency. Between four and nine clinical lots were assessed for various parameters such as relative formulation of collagenase isoforms, and recovered collagenase and protease potencies postreconstitution. RESULTS: Six vials from a single typical lot had a mean enzyme content of 489+/-62.5 mg (mean+/-SEM; range 398-610 mg). The mean recovered collagenase activity was 2235+/-310 Wunsch units (WU)/vial (range 1794-2968 WU/vial). The percent coefficients of variation for collagenase and protease activity in these vials were 17.4%, and 13.4%, respectively. The increase in the presence of the collagenase Ib (CIb) isoform detected by HPLC analysis was related to the chronological order of the date of manufacture. The CIb isoform was found to have a reduced specific activity compared to intact collagenase I (CI) (3.8+/-1.2 WU/mg vs. 2.1+/-0.7 WU/mg, P < 0.05). The presence of CIb was related to reduced islet yields in twelve human isolations studied. CONCLUSIONS: Variation in potency was observed between, and within lots of Liberase HI in this study. Differences in relative collagenase isoform composition may also affect the stability and potency characteristics of these blends. PMID- 16210958 TI - Validation of large particle flow cytometry for the analysis and sorting of intact pancreatic islets. AB - BACKGROUND: Accurate quantification of total islet yield is an essential step prior to transplantation and for research. The standard method of manually determining an islet equivalent (IEQ) count is subjective and prone to error. We evaluated Complex Object Parametric Analyzer and Sorter (COPAS) large particle flow cytometry for the determination of islet equivalent counts and purities of islet preparations. METHODS: Initial validation of the sensitivity and accuracy of the COPAS flow cytometer was performed by analysis and sorting of uniform polystyrene microspheres with sizes similar to islets. Human and Rhesus monkey islets were stained with the zinc-specific fluorescent dye Newport Green to discriminate islet from nonislet tissue. Islet sizes were extrapolated from standard curves obtained using microspheres from which individual islet volumes were calculated. IEQ counts on six islet preparations were performed by the standard manual method and compared with results obtained by automated COPAS flow cytometry. RESULTS: The COPAS flow cytometer was highly accurate in the detection and measurement of both polystyrene microspheres and islets. IEQ counts determined by COPAS flow cytometry were consistent with manual counts although subject to error when assessing preparations with significant numbers of islets embedded within acinar tissue. Size-specific islet sorting with retention of morphology and dithizone staining was also shown using the COPAS flow cytometer. CONCLUSIONS: COPAS large particle flow cytometry provides a novel automated approach for quantification of intact islets and determination of islet equivalent yield. In addition, the ability to analyze and sort islets based upon user defined criterion opens unique avenues for experimentation. PMID- 16210959 TI - Possibility of islet transplantation from a nonheartbeating donor pancreas resuscitated by the two-layer method. AB - BACKGROUND: The shortage of cadaveric donors is a problem in islet transplantation, and recent improvements in this field have led to renewed interest in the use of nonheartbeating (NHB) donors. NHB donor pancreata that could provide a significant source for islet transplantation are associated with warm ischemic injury. We tested whether the two-layer method (TL) could improve islet yield and function from damaged pancreata after warm ischemia (WI). METHODS: Lewis rats were divided into six groups. In groups 1 to 3, rats were subjected to 0, 30, and 45 minutes of WI, respectively. Islets were isolated immediately (subgroup a) or after 3-hour preservation with TL (subgroup b). Isolated islets were assessed in terms of islet yield and in vivo function. We also assessed the pancreatic tissue ATP concentration before isolation and distended pancreata morphologically after chemical digestion by H&E staining. RESULTS: Islet yield decreased significantly after 30 minutes of WI in group 2a, whereas TL preservation doubled this decreased yield in group 2b. Forty-five minutes of WI resulted in nearly no islet yield in both groups 3a and 3b. The success rates of transplantation in groups 1a, 1b, 2a, and 2b were 100%, 100%, 0%, and 75%, respectively. Increased tissue ATP levels and alleviation of morphological islet damage were observed in group 2b. CONCLUSIONS: These results demonstrated that pancreata damaged from 30-minute WI were restored by 3-hour TL preservation. TL may allow the selective use of NHB donors as an alternative source for islet transplantation. PMID- 16210960 TI - Cyclosporine formulation and Kaposi's sarcoma after renal transplantation. AB - BACKGROUND: Transplantation enhances the risk of malignancies, due to the chronic use of antirejection medication. In the case of Kaposi's sarcoma (KS) the permissive effect of immunosuppression has been extensively studied, and cyclosporine (CsA) appears to play a key role. Here we have compared the incidence of KS in transplant patients receiving Neoral or Sandimmune as a part of their immunosuppressive therapy. METHODS: In all, 668 kidney transplant recipient followed at our Nephrology Unit from 1970 to 2003 entered this retrospective analysis; 300 were on CsA Sandimmune-based and 308 on CsA Neoral based therapy. The primary endpoint was the occurrence of KS. RESULTS: KS was diagnosed in 20 out of 608 patients given CsA with an incidence rate of 4.7 per 1000 patients per year. No episodes of KS was found in the preCsA era. Among patients on CsA, those treated with Neoral had fourfold higher incidence rate of KS than in the Sandimmune group (10.7 vs. 2.3 per 1000 patients per year). Kaplan Meier analysis shows that patients on Neoral had lower cumulative KS-free probability than those on Sandimmune. Cox's analysis documented that Neoral was a positive predictor of KS development as compared to Sandimmune (hazard ratio: 2.237). Among patients on Neoral, those who developed KS had higher daily exposure to the drug assessed by pharmacokinetic studies. CONCLUSIONS: In recipients of kidney transplant CsA Neoral increases the risk of KS as compared to the Sandimmune formulation, possibly due to enhanced drug bioavailability and ultimately patients daily CsA exposure. PMID- 16210961 TI - Low incidence of malignancy among sirolimus/cyclosporine-treated renal transplant recipients. AB - BACKGROUND: Malignancies, a well-known complication of immunosuppressive therapy in renal transplant recipients, represent an important cause of long-term morbidity and mortality. One approach to addressing this problem is identifying agents that display antineoplastic properties concomitant with their immunosuppressive effects. METHODS: We examined the neoplasms among 1008 renal transplant recipients treated at a single center with sirolimus-cyclosporine +/- prednisone. RESULTS: Clinical and laboratory data, including 62.3+/-26.1 months follow-up (range 27.1-131), revealed 36 tumors in 35 patients (3.6%) presenting at 32.5+/-29.8 months. The 2.4% incidence of skin tumors, the most common neoplasms, was 1.58-fold greater than the general U.S. population. In addition to a 0.4% incidence of posttransplant lymphoproliferative disorders (PTLD) and a 0.2% incidence of renal cell carcinomas, we observed single cases of breast, bladder, endometrial, lung, and brain neoplasms as well as leukemia. The mean trough drug concentrations at the time of diagnosis in affected recipients were within our putative target ranges. In addition to eleven graft losses due to death with a functioning kidney, two were related to chronic rejection following reduced immunosuppression, and one, therapeutic nephrectomy for PTLD. Five of twelve deaths were caused by malignancies; four others among 1008 patients over the entire follow-up were attributed to cardiovascular events; one, to respiratory failure; and two, at distant locations to unknown causes. CONCLUSIONS: The sirolimus-cyclosporine +/- prednisone combination appears likely to be associated with a reduced incidence of tumors. PMID- 16210962 TI - Incidence of cancers following orthotopic liver transplantation in a single center: comparison with national cancer incidence rates for England and Wales. AB - BACKGROUND: The incidence of de novo cancers is increased in liver allograft recipients but there are few data assessing the extent of the increased risk compared with a matched population. METHODS: A retrospective study of 1,778 adults transplanted between January 1982 and March 2004, followed for a median of 65 months. The observed cancer incidence was compared with age-, sex-, and calendar year-matched expected cancer rates in England and Wales population. RESULTS: In all, 141 (7.9%) developed a new cancer. There was an increase in the incidence of all tumors compared with that expected (Standardized Incidence Ratio (SIR) 207, 95% CI 174-244, P < 0.001); the greatest increase was seen in lymphoid tumors (SIR 1026, 95% CI 608-1621, P < 0.001), skin cancers (SIR 580, 95% CI 432 763, P < 0.001), and cancer of the large bowel (SIR 496, 95% CI 290-774, P < 0.001). Large bowel cancer was more common in those patients with ulcerative colitis than those without (SIR 2727 vs. 347) and in older patients. Females had a greater risk of lung cancer than males (SIR 336 vs. 56). CONCLUSIONS: There is an increased incidence of tumors following liver transplantation. Although the absolute risk of cancer is low, we found that the increase in risk is greater in the younger aged recipients than the older ones. Increased awareness of colon cancer is needed especially in older patients and those with ulcerative colitis. There should be awareness for the high lung cancer incidence in females. Increased surveillance for breast and cervical cancer is not necessary. PMID- 16210963 TI - Randomized trial of Alemtuzumab for prevention of graft rejection and preservation of renal function after kidney transplantation. AB - BACKGROUND: A randomized, multicenter, controlled trial was undertaken to evaluate the safety and efficacy of Alemtuzumab, a powerful lytic agent for both T and B lymphocytes, in the prophylaxis of rejection in renal transplantation (RTx). METHODS: Thirty patients were randomized to receive Alemtuzumab together with low-dose cyclosporine (CsA) monotherapy (CAMPATH, n = 20) or to full doses of CsA with azathioprine and corticosteroids (Standard, n = 10). CsA was administered at doses to achieve whole-blood trough CsA levels of 90 to 110 ng/mL and 180 to 225 ng/mL in CAMPATH and Standard groups, respectively. RESULTS: Per protocol, CsA trough levels were lower in patients assigned to CAMPATH post-RTx (median trough level of 119 vs. 166 ng/mL at 6 months, CAMPATH vs. Standard; 95% confidence interval, -92 to -34). At 6 months post-RTx, serum creatinine, graft and patient survivals, incidence of biopsy proven acute rejection (25% vs. 20%, CAMPATH vs. Standard), overall treatment failure, and severe and moderate infections were comparable. Whereas all patients receiving Standard therapy required maintenance corticosteroids at 6 months, of the 17 of 20 patients with functioning grafts in CAMPATH, 15 (88%, 95% confidence interval, 53%-97%) were steroid free. CONCLUSION: These results suggest that Alemtuzumab is an effective induction agent that permits low-dose steroid-free immunosuppression in RTx. PMID- 16210964 TI - Home glucometer monitoring markedly improves diagnosis of post renal transplant diabetes mellitus in renal transplant recipients. AB - BACKGROUND: Definitions of de novo posttransplant diabetes mellitus (PTDM) have varied widely in the renal transplant literature, and most have not used the American Diabetes Association (ADA) definition of diabetes (fasting plasma glucose [FPG] > or = 126 mg/dl on two occasions, or a casual plasma glucose level >200 mg/dl). Most patients are monitored for PTDM by 12-hour FPG levels drawn for clinic visits. In contrast, we describe the diagnosis of PTDM by home glucometer monitoring METHODS: We screened 89 consecutive nondiabetic renal transplant recipients for PTDM by ADA criteria and home glucometer monitoring during the first 3 months posttransplant RESULTS: Of 23 patients with impaired fasting glucose levels of 111-126 mg/dl, 14 (61%) met ADA criteria for diabetes mellitus of based on home glucometer monitoring. The incidence of de novo PTDM was 31% during this period. Predictors of PTDM in a Cox proportional hazards model were race and acute rejection, with a trend towards BMI. Clinic visit FPG levels did not differ between PTDM and non-PTDM patients. All diagnoses were made based on prelunch or supper FPG >200 mg/dl. CONCLUSIONS: Overnight FPG are inadequate for diagnosis of PTDM. All renal transplant recipients with impaired FPG should, at minimum, have home FPG testing. PMID- 16210965 TI - Inolimomab in steroid-refractory acute graft-versus-host disease following allogeneic hematopoietic stem cell transplantation: retrospective analysis and comparison with other interleukin-2 receptor antibodies. AB - BACKGROUND: The use of monoclonal antibodies against interleukin-2 receptor (IL 2R)-alpha chains could be an effective treatment of acute graft-versus-host disease (GvHD). Experimental model and clinical studies have reported various results. METHODS: Inolimomab is a murine anti-IL-2R. Eighty-five patients were evaluated retrospectively for the safety and efficacy of inolimomab given for the treatment of steroid-resistant acute GvHD (aGvHD) following allogeneic hematopoietic stem cell transplantation (HSCT). Diseases were immune deficiency, hematological malignancies, or solid tumors. Seventy-six percent of the patients received a myeloablative regimen. The source of HSCT was bone marrow for 45 patients, peripheral blood for 36 patients, and cord blood for 4 patients. Donors were 49 siblings and 36 unrelated. Acute GvHD was diagnosed within a median of 28 days after transplantation (grade II, 26 patients; grade III, 26 patients; grade IV, 33 patients). Inolimomab was administered in the event of steroid-resistant aGvHD with a median dose of 0.468 mg per kg (median period of treatment: 18 days). RESULTS: Twenty-five complete responses and 29 partial responses (total response rate: 63%) were observed with no side effects. There was no correlation between aGvHD grading and quality of response. Better responses were observed in cutaneous aGvHD. The overall survival probability was 26% (median follow-up: 20 months). Fifty-seven percent of patients died of toxicity related mortality, mostly aGvHD. Response to inolimomab seemed sustained (11% relapse in responders). CONCLUSION: Inolimomab is well-tolerated and effective for severe steroid-resistant aGvHD. The optimum regimen remains to be defined. PMID- 16210966 TI - High rate of recurrence in renal transplant recipients after a first episode of venous thromboembolism. AB - BACKGROUND: No data are available about the optimal duration of oral anticoagulant therapy (OAT) after an episode of venous thromboembolism (VTE) occurring in renal transplant (RT) recipients. Our study was undertaken to evaluate the risk of VTE recurrence in patients developing a first episode of VTE after RT. METHODS: Among 484 RT patients, 34 (7%) developed a first VTE: 28/34 VTE patients (Group 1) were prospectively studied, after stopping OAT. Group 1 was compared with a group of 84 patients without history of renal disease who had suffered from a first episode of VTE matched for age, sex and type of thrombotic event (Group 2) and with a matched group of 84 RT recipients with no history of VTE (Group 3). After OAT withdrawal, blood samples were obtained for thrombophilia and clotting activation markers (prothrombin fragment 1+2 (F1+2) and D-dimer plasma levels). RESULTS: During follow-up, 14/28 patients of Group 1 and 8/84 patients of Group 2 experienced VTE recurrence (P < 0.0005). Homocysteine, F1+2 and D-dimer plasma levels were significantly higher in Group 1 than in Group 2 and 3 (P < 0.0001 and <0.05 respectively) for all the three parameters. CONCLUSIONS: Our data outline the high risk of VTE recurrence in RT recipients. Strategies for VTE recurrence prevention are needed; Prolonged OAT, in spite of the high bleeding risk of RT patients, should be considered in this respect. PMID- 16210967 TI - Excellent outcomes after transplantation of deceased donor kidneys with high terminal creatinine and mild pathologic lesions. AB - BACKGROUND: Outcomes after kidney transplantation using deceased donors with high terminal creatinine are not well described but potentially represent an underutilized source of renal allografts. Utility of renal biopsy of these kidneys is similarly not well established. METHODS: To better understand the posttransplant function of kidneys from donors with high terminal creatinine, we reviewed our database of almost 500 cadaveric kidney transplants. We compared the 65 nonexpanded criteria donors with a final donor creatinine > or = 2.0 mg/dl (range 2.0-4.9 mg/dl) with kidneys procured from donors with terminal creatinine of <1.5. Biopsy results were correlated with graft function. RESULTS: Kidneys from deceased donors with high terminal creatinine performed as well as kidneys from donors with a normal terminal creatinine with respect to primary non function, acute rejection, 6-year graft and patient survival, and function over the first 48 months. High creatinine kidneys with moderate or severe lesions on biopsy demonstrated poorer function at 6 months and 1 year as compared to those with mild or no histological lesions. CONCLUSIONS: Under select conditions, kidneys from donors with high terminal creatinine can be used safely with excellent results. PMID- 16210968 TI - The standardization of pancreatic donors for islet isolations. AB - BACKGROUND: Islet transplantation has proven to be a successful treatment for type 1 diabetes mellitus. The aim of this study was to establish an algorithm by which the combination of the donor quality and pancreas quality was given a numerical score from 0 to 100 for use in determining the quality of a pancreas for islet isolation. METHODS: We retrospectively analyzed 326 pancreases and the outcomes of their respective isolations. Specific donor variables and physical characteristics were identified and weighted according to their influence on the success of the isolation. For each variable, ranges and point weightings were established based on our laboratory experience and literature review. RESULTS: Analysis of the data showed a strong association of the donor point with isolation outcome. Pancreases with lower donor point scores had lower transplant success rates, whereas higher donor point scores in turn produced higher transplant rates. CONCLUSIONS: This scoring system has proven to be useful in guiding the decision process as to whether to accept a particular pancreas for a favorable isolation outcome. PMID- 16210970 TI - Risk-stratified screening for ischemic heart disease in kidney transplant candidates. AB - BACKGROUND: Few studies have examined the effectiveness of a risk-stratified approach to screening kidney transplantation candidates for ischemic heart disease (IHD). METHODS: We retrospectively reviewed records from all adult patients (n = 514) placed on the deceased donor kidney transplantation waiting list at a single center between January 1992 and June 2000. During this time there was a consistent policy for high-risk patients to undergo noninvasive stress testing and/or coronary angiography. We examined screening tests, the resulting interventions, and the incidence of subsequent IHD events (myocardial infarction, angioplasty, bypass surgery, or IHD death) among those screened and not screened. RESULTS: For 224 (43.6%) low-risk patients who were not screened, the actuarial incidence of an IHD event after listing (before or after transplantation) was only 0.5% at 1 year, 3.5% at 3 years, and 5.3% at 5 years. Screening 290 (56.4%) high-risk patients resulted in prophylactic angioplasty in 18 (6.2%), and bypass surgery in 8 (2.8%) before listing. After listing, 61 patients were screened, resulting in angioplasty in 6 (9.8%) and bypass surgery in 1 (1.6%). Of the 68 patients who ultimately had an IHD event after being placed on the waiting list, only 13 (19.4%) had not been screened. CONCLUSIONS: A risk-stratified screening strategy effectively avoided unnecessary testing in 43.6%. However, the relatively low proportion of screened patients who underwent prophylactic angioplasty or bypass grafting raises the question of whether screening was effective in preventing IHD events. PMID- 16210969 TI - Comparison of steroid avoidance in tacrolimus/mycophenolate mofetil and tacrolimus/sirolimus combination in kidney transplantation monitored by surveillance biopsy. AB - BACKGROUND: Chronic steroid therapy in kidney transplantation has myriad side effects and steroid avoidance has become feasible. This prospective study compared the safety and efficacy of steroid avoidance in tacrolimus (TAC)/mycophenolate mofetil (MMF) and TAC/sirolimus (SRL) combinations in kidney transplantation. METHODS: In all, 150 kidney recipients were analyzed: 75 each in TAC/MMF and TAC/SRL groups. The primary endpoint was acute rejection. Surveillance biopsies were completed to analyze subclinical acute rejection (SCAR) and chronic allograft nephropathy (CAN). Acute rejection and SCAR were treated by methylprednisolone. Two-year patient and graft survival, renal function, and adverse effects were monitored. RESULTS: Acute rejection was seen in 12% of TAC/MMF and 8% of TAC/SRL patients. Two-year actuarial patient survival was 95% and 97%, and graft survival 90% and 90% in TAC/MMF and TAC/SRL groups, respectively. Surveillance biopsy showed cumulative incidence of SCAR was 27 % in TAC/MMF and 16 % in TAC/SRL groups at 2 years (P = 0.04). Overall, 33% of recipients in TAC/MMF and 20% in TAC/SRL received methylprednisolone for acute rejection/SCAR. Moderate/severe CAN was 10% in TAC/SRL group and 22% in TAC/MMF group(P = 0.06). New-onset diabetes mellitus (NODM) was 4% each in both groups. All recipients remain free of maintenance steroid therapy. CONCLUSIONS: Steroid avoidance in tacrolimus-based immunosuppression with MMF or SRL provides equivalent 2-year patient and graft survival with a low incidence of acute rejection and NODM. SCAR and CAN are lower in TAC/SRL compared to TAC/MMF group. The impact of decreased SCAR and CAN in TAC/SRL group on longer-term graft survival and function is to be evaluated. PMID- 16210971 TI - Prolonged survival of microencapsulated neonatal porcine islets in mice treated with a combination of anti-CD154 and anti-LFA-1 monoclonal antibodies. AB - BACKGROUND: The aim of this study was to determine whether short-term administration of a combination of anti-CD154 and anti-LFA-1 monoclonal antibodies can prolong the survival of microencapsulated neonatal porcine islets (NPI) in immunocompetent mice. METHODS: Microencapsulated NPI were transplanted into the peritoneal cavity of streptozotocin-induced diabetic B6 mice that received a short-term treatment of a combination of anti-CD154 and anti-LFA-1 monoclonal antibodies. Blood glucose levels of each recipient were measured for more than 100 days posttransplantation or until graft rejection. Microcapsules were recovered to determine the presence of immune cells using immunoperoxidase staining. In addition, the levels of mouse anti-porcine immunoglobulin (Ig) G antibodies in the serum of each recipient were measured by flow cytometry. RESULTS: Short-term administration of a combination of monoclonal antibodies resulted in significant prolongation of microencapsulated NPI xenograft survival. All treated mice (n = 20) achieved normoglycemia within 10-35 days posttransplantation and 11/20 mice remained normoglycemic for more than 100 days posttransplantation. In contrast, only 1/20 of the untreated mice achieved normoglycemia and this mouse became diabetic at 17 days posttransplantation. Histological examination of the recovered microcapsules from long-term surviving treated mice revealed minimal cellular overgrowth containing intact viable islets, whereas several layers of immune cells surrounding the capsules containing nonviable islets were observed in untreated mice. The levels of mouse anti-porcine IgG was also reduced in treated recipients compared to untreated mice. CONCLUSIONS: These data demonstrate that short-term administration of anti CD154 and anti-LFA-1 monoclonal antibodies can be effective in promoting long term survival of microencapsulated NPI in immune-competent mice. PMID- 16210972 TI - Disruption of P-selectin signaling modulates cell trafficking and results in improved outcomes after mouse warm intestinal ischemia and reperfusion injury. AB - BACKGROUND: This study analyzes the role of T lymphocytes and neutrophils (PMN) in intestinal ischemia and reperfusion injury (IRI) using either P-selectin blockade or elimination. METHODS: Using a model of severe mouse warm intestinal IRI, the following groups were performed: group 1: wild type C57BL6 no treatment; group 2: wild type treated with r-PSGL1-Ig; group 3: C57BL6 genetically deficient in P-selectin. Survival was assessed at day 7; intestine was assayed for histopathology, apoptosis, myeloperoxidase (MPO), inflammatory cytokines, hemoxygenase-1 (HO-1), and CD3 lymphocytes. Standard statistical comparison was undertaken. RESULTS: The survival was significantly (P < 0.01) improved in the treatment groups: group 1, 50%; group 2, 90%; group 3, 100%. Graded histopathology and crypt apoptosis were improved in groups 2 and 3. MPO and CD3 positive cells were significantly reduced in groups 2 and 3. A significant reduction in inflammatory/Th1-type cytokines was seen in groups 2 and 3 as compared to group 1. Conversely, a significant increase in Th2-type cytokines and HO-1 production was seen selectively in groups 2 and 3. CONCLUSIONS: This study demonstrates the importance of P-selectin signaling in warm, murine intestinal IRI in that either the blockade of or the genetic deficiency in P-selectin confers a survival advantage and reduction in tissue injury/inflammation. The mechanism involves a reduction of PMN and CD3 T cell infiltration and an alteration in the cytokine microenvironment in favor of a Th2 profile. These data implicate T lymphocyte as an important regulatory cell in this inflammatory process. PMID- 16210973 TI - Multipotent mesenchymal stem cells with immunosuppressive activity can be easily isolated from dental pulp. AB - BACKGROUND: Bone marrow mesenchymal stem cells (MSCs) are currently being investigated in preclinical and clinical settings because of their multipotent differentiative capacity or, alternatively, their immunosuppressive function. The aim of this study was to evaluate dental pulp (DP) as a potential source of MSCs instead of bone marrow (BM). METHODS: Flow cytometric analysis showed that DP MSCs and BM-MSCs were equally SH2, SH3, SH4, CD29 and CD 166 positive. The in vitro proliferative kinetics of MSCs were measured by 3H-thymidine incorporation uptake. The immunosuppressive function of MSCs was then tested by coculturing PHA stimulated allogeneic T cells with or without MSCs for 3 days. RESULTS: BM-MSCs could be differentiated in vitro into osteogenic, chondrogenic and adipogenic lineages. DP-MSCs showed osteogenic and adipocytic differentiation, but did not differentiate into chondrocytes. Although DP-MSCs grow rapidly in vitro between day 3 and day 8 of culture and then decrease their proliferation by day 15, BM MSCs have a stable and continuous proliferation over the same period of time. The addition of DP-MSCs or BM-MSCs resulted in 91 +/- 4% and 75 +/- 3% inhibition of T cell response, respectively, assessed by a 3H-thymidine assay. CONCLUSIONS: Dental pulp is an easily accessible and efficient source of MSCs, with different kinetics and differentiation potentialities from MSCs as isolated from the bone marrow. The rapid proliferative capacity together with the immunoregulatory characteristics of DP-MSCs may prompt future studies aimed at using these cells in the treatment or prevention of T-cell alloreactivity in hematopoietic or solid organ allogeneic transplantation. PMID- 16210974 TI - Peripheral dendritic cell chimerism in allogeneic hematopoietic stem cell recipients. AB - BACKGROUND: Relapse and graft-versus-host disease (GVHD) represent major causes of morbidity and mortality following allogeneic hematopoietic stem cell transplantation (HSCT). Although leukocyte and T-cell chimerism analyses are performed routinely suggesting a predictive value on the patients outcome, little is known about chimerism of dendritic cells (DC) representing strong initiators of immune responses. METHODS: In this prospective study, peripheral DC1 (CD11c+) and DC2 (CD123+) chimerism was determined in hematopoetic stem cell recipients. DCs were isolated from peripheral blood by fluorescence activated cell sorting. Chimerism analyses were performed by fluorescent in situ hybridization or by polymerase chain reaction-based typing of short tandem repeats. RESULTS: At time of engraftment, DC chimerism analyses showed complete chimerism in 76.3% (DC1)/79.5% (DC2), mixed chimerism (MC) in 21.0% (DC1)/17.9% (DC2) and no chimerism in 2.7% (DC1)/2.6% (DC2) of the patients. Peripheral DC chimerism had no significant effect on relapse-free or overall survival. Although acute GVHD was observed more often in patients with MC for DC1/DC2 and chronic GVHD occurred more often in patients with MC for DC2, there was no statistically significant correlation. CONCLUSIONS: Although DCs as antigen presenting cells are supposed to have an impact on the induction of GVHD, there was no significant correlation between incidence of GVHD and DC chimerism after HSCT. PMID- 16210975 TI - Naive CD4+ cells from cord blood can generate competent Th effector cells. AB - BACKGROUND: Umbilical cord blood (UCB) cells have been increasingly used as a source of hematopoietic stem cells for allogeneic transplantation. Previous reports suggest that the low risk of graft-versus-host disease in patients that received cord blood cells seems related to the distinctive nature of cord blood T cells. METHODS: To analyze the maturation of CD4+CD45RA+ cord blood cells, we performed an in vitro differentiation assay to compare the generation of Th effector cells strictly from UCB and adult peripheral blood (APB) CD4+CD45RA+ cells. RESULTS: During the maturation into effector cells, UCB and APB cells acquired a comparable activation level determined by the expression pattern of CD69, CD40L, OX40 and CD62L as well as PD1 and CTLA-4 molecules. Moreover, the expression of CD45RO isoform was induced in most activated effector cells from both UCB and APB. OKT3-restimulated effector cells generated from naive UCB expressed higher levels of CD25 coinciding with the secretion of higher amounts of IL-2. Effector cells from both origins consisted of heterogeneous populations with similar frequencies of Th1 and Th2 cytokine producing cells, secreting equivalent levels of IL-4, IL-5 and IFNgamma. Although, higher levels of IL-10 were detected in the cytokine mRNA profile and in the supernatants of OKT3 restimulated UCB effector cells, blocking endogenous IL-10 with anti-IL-10 mAbs enhanced significantly the proliferative response of UCB as well as APB effector cells (P < 0.05). CONCLUSIONS: These results indicated that Th effector cells generated from naive UCB cells were intrinsically as competent as naive APB to respond to TCR-mediated stimulation. In addition, UCB effector cells produced higher IL-10 but its inhibitory effect on proliferation may be partially compensated by the higher production of IL-2 and enhanced expression of CD25. PMID- 16210977 TI - Nephron-sparing surgery for de novo renal cell carcinoma in allograft kidneys. AB - Renal cell carcinomas account for 4.6% of post-transplant cancers, 10% of which occur in allograft kidneys. We report three such cases among kidney grafts that were performed or followed from 1970 to 2004. In all patients, we performed a partial allograft nephrectomy after consideration of the tumor size, location, and absence of metastases and local extension. Renal function has remained stable, and there has been no sign of graft rejection, tumor recurrence or metastases. The surgery was technically feasible without exposing the patients to increased postoperative risks. The lateral, peripherally located tumor allowed excision without renal hilar dissection or entry into the collecting system. In agreement with data emerging from the literature, the present cases confirm that even in the setting of long-standing immunosuppression, de novo RCC of the kidney graft warrants a minimally invasive approach to spare patients graft loss and return to hemodialysis. PMID- 16210976 TI - Long-term outcome of adding mycophenolate mofetil to tacrolimus for nephrotoxicity following liver transplantation. AB - Mycophenolate mofetil (MMF) has no known nephrotoxicity. This report examines the outcome in patients who received MMF for renal impairment on tacrolimus-based immunosuppression. From 1995 to 1996, twelve liver transplantation (LTx) patients (mean age 54.6 years) with serum creatinine >1.8 mg/dl were included in the study. MMF was introduced and tacrolimus dose was reduced by 30-50%. Each patient was followed for 6 years. Renal function showed improvement in seven patients, deterioration in four, and no change in one patient. Overall mean serum creatinine decreased from 2.5 to 1.9 mg/dl at 6 months but increased to 2.2 mg/dl at 18 to 24 months. After that, renal function remained stable for 72 months. Iothalamate clearance showed 18.5% improvement at 1 year. Three patients developed renal failure. Six patients died in the follow-up period. Addition of MMF with reduced tacrolimus dose resulted in sustained improvement in renal function in 58% of patients. PMID- 16210978 TI - Candida glabrata fungemia in transplant patients receiving voriconazole after fluconazole. AB - The clinical impact of voriconazole resistance in Candida glabrata is not well described. Five hematopoietic stem cell transplant recipients that developed breakthrough Candida glabrata bloodstream infections while receiving voriconazole are described and the clinical management and susceptibility profiles of their isolates are reported. All patients were markedly immunosuppressed, and in all cases, voriconazole use was preceded by prolonged fluconazole exposure (median 60 days); median voriconazole exposure prior to candidemia was 48 days. Isolates from 4 patients were shown to be resistant to fluconazole and itraconazole when tested in vitro; these same isolates had MICs to voriconazole and posaconazole > or = 2 microg/ml. Clinical failure to voriconazole may result from deficits in host defense, retained infected foci, and adaptation of the organism to environmental pressures, the specific sequence and mechanisms of which warrant further study. Clinicians must maintain a high index of suspicion for these infections in highly susceptible hosts despite voriconazole therapy, particularly when voriconazole use is preceded by prolonged fluconazole exposure. PMID- 16210979 TI - Recipient age and mycophenolate mofetil as the main determinants of outcome after steroid withdrawal: analysis of long-term follow-up in renal transplantation. AB - The long-term benefit of steroid withdrawal on patient and graft survival is unproven. Steroids were stopped within the first year after kidney transplantation in 223 consecutive low-risk patients initially treated with thymoglobulin and triple-drug therapy. The 15-year actuarial graft survival rate was 83.9%. Risk factors for graft loss were: proteinuria (hazard ratio [HR]: 6.96, P = 0.0003), creatinine >130 micromol/L (HR: 3.37, P = 0.01), recipient age <35 years (HR: 5.31, P = 0.001), and no mycophenolate mofetil (MMF) treatment (HR: 8.83, P = 0.04). Interestingly, recipient age and no MMF treatment were not risk factors in higher-risk patients in whom steroids were continued. The 2-year incidence of acute rejection following steroid withdrawal was 12.1%; the graft survival rate was lower in this group (71.1%). Our findings indicate that late steroid withdrawal results in excellent long-term outcome in most low-risk patients, but it should be attempted with caution in younger patients and when MMF is not used. PMID- 16210980 TI - A pilot study to assess the use of digital photography in renal transplantation. PMID- 16210982 TI - Image-guided laparoscopic surgery. Review and current status. AB - The main drawback with laparoscopic surgery is that the surgeon is unable to palpate vessels, tumours and organs during surgery. Further-more, the laparoscope only provides a surface view of organs. There is a need for more advanced visualizations techniques that can enhance the display presented to the surgeon so that important information below the surface of the organs is included when planning the procedure as well as for guidance and control during treatment. In this paper, we present a review of the literature and the state of art within image-guided laparoscopic surgery. We describe our own experience using a prototype navigation system for advanced visualizations and guidance during laparoscopic procedures in the retroperitoneum. Furthermore, we show sample images from the Future Operating Room for laparoscopic surgery in Trondheim, where this technology is being further developed and tested in clinical studies. Our system is based on three-dimensional navigation technology, i.e. preoperatively acquired magnetic resonance or computed tomography data used in combination with tracked instruments, allowing the surgeon to interactively control the display of images prior to and during surgery with normal use of the instruments. In summary, we believe that abdominal image navigation using tracked instruments and advanced visualizations has a large potential for improving future laparoscopic surgery, especially in cases where vessels and anatomical relations beyond surfaces is difficult to identify using only a laparoscope. The technology helps the surgeon to better understand the anatomy and locate blood vessels. Accordingly, we believe that this new technology could increase safety and make it easier for the surgeon to perform successful laparoscopic surgery. PMID- 16210983 TI - Laparoscopic esophagectomy. AB - Minimally invasive esophagectomy is emerging as an alternative option to open esophagectomy for benign and malignant esophageal diseases. This article provides a detailed review of the history of minimally invasive esophagectomy and an update on the currently accepted techniques for minimally invasive esophagectomy and its outcomes. PMID- 16210984 TI - Laparoscopic resection of colorectal cancer. AB - Laparoscopic surgery has revolutionised procedures such as cholecystectomy since its inception in the 1980s. After initial enthusiasm with laparoscopic colorectal resections in the early 1990s, resection of colorectal malignancy was largely abandoned outside clinical trials because of reports of inferior oncological outcomes including local and port-site recurrence. More recently, however, an increasing number of reports have demonstrated that laparoscopic surgery for colorectal cancer though technically demanding is feasible, and the results of large multi-centred randomised trials showing oncological equivalence are becoming available. Technological advances in laparoscopic equipment along with the increasing skills and experience of laparoscopic surgeons have extended the indications and reduced the contraindications for laparoscopic colectomy. This, along with the use of fast- track protocols is changing the way we manage patients. The future of laparoscopic colorectal surgery is assured, driven not only by the physical benefits to the patient in the short and medium term, the reduced financial burden on in-patient stay, and post-operative return to work, but also increasing patient demand. This in turn requires that surgeons should ensure high quality training and operative competence to maintain the high standards achieved by the pioneers in this field. PMID- 16210985 TI - Laparoscopic radical prostatectomy: an evolution in surgery. AB - The late 1990s witnessed an unprecedented evolution in the surgical approaches to the prostate thru the eye of the laparoscope. Initially taken with doubt, laparoscopic radical prostatectomy (LRP) has gained tremendous popularity and widespread implementation at specialized centers worldwide becoming the standard in many of them. LRP represents a technically demanding laparoscopic procedure but it can be performed systematically with standard techniques. Obvious advantages are shorter convalescence and markedly lower operative blood loss without compromise of cancer control. Long-term functional and oncological results are maturing but early reports of positive surgical margin rates and freedom from prostate-specific antigen (PSA) recurrence rates after LRP are encouraging. Early quality of life results of postoperative urinary and sexual function appear similar to those in open surgical series. The real challenge for laparoscopic surgeons entails a paradigm swift, one that breaks off from the traditional and bias impetus, and tests the instruments, procedures, techniques differences and evaluates outcomes in a prospective controlled and randomized manner. If achieved, the laparoscopic movement may give rise to a generation of forward thinking surgeons generating a wealth of clinical evidence for their patients. PMID- 16210986 TI - Laparoscopic splenectomy for non-traumatic diseases. AB - At present, laparoscopic splenectomy (LS) is mostly indicated in hematologic benign diseases, and in case of normal size spleen it is considered the gold standard. The technique is under a continuous evolution and several studies have demonstrated feasibility and safety of laparoscopic approach also in case of massive splenomegaly, malignant diseases and even in the treatment of selected cases traumatic lesions. LS is an advanced surgical procedure that requires a management of a full trained team involved in the preoperative preparation, surgical strategy and postoperative care. A fully comprehension of the impact of the surgical strategy is needed to plan the treatment of diseases that often are managed in a multidisciplinary setting. Indications and contraindications to surgery does not differ significantly from open traditional splenectomy, but the nature of the disease and the volume of the spleen can greatly influence the operation. A preoperative study with definition of these parameters along with a standard preparation to surgery is required. Over the years the technique has been developed and adapted to respond to emergent necessities related to those parameters. Anterior, semilateral, later and hand assisted approaches offer advantages and drawbacks that should be weighed in each case. They depend on surgeons preferences but most of the time are related to the preoperative studies, even when new surgical tools (i.e. harmonic scalpel and radiofrequency). No randomized, prospective trials have been conducted, however several studies with strong evidence have shown that less surgical trauma, intraoperative blood loss, early hospital discharge, rapid return to normal activities and better cosmesis can be obtained with a laparoscopic approach. PMID- 16210987 TI - Hepatic resection for colorectal metastasis; time to challenge the accepted doctrine. AB - The selection of patients for resection of colorectal liver metastasis (CRLM) is based around a set of established rules and principles, some of which date back to and have changed little since the mid 1980's. In this paper the authors challenge this accepted doctrine and describe the criteria used for selection of patients for surgery in their own centre, criteria which permit the inclusion of many more patients for potentially curative surgery. They go on to describe methods used to increase resectability and discuss their own results achieved for the resection of CRLM. PMID- 16210988 TI - Venous thromboembolism in morbid obesity and trauma. A review of literature. AB - Deep venous thromboembolism (DVT) is common and leads to disability, economic loss and even death. The aims of this paper are to start from the basic knowledge that we have about DVT and to tailor our knowledge to the treatment and diagnosis of thromboembolism in obese patients and trauma patients, which are among the patients who have a high risk of developing DVT and pulmonary embolism. Venous thromboembolism is a common complication in patients with major trauma, and effective, safe prophylactic regimens are needed. The patients are treated effectively with heparin and low molecular weight heparins, which are shown to be safe and effective. Morbid obesity is a disease that affects 10% of Americans and increases the incidence of DVT. Forty mg of enoxaparin subcutaneously twice a day seems to be a better DVT prophylaxis than the 30 mg twice a day. Many patients admitted to the hospital are morbidly obese; therefore, we suggest they start on low molecular heparin. The high morbidity of these patients is because most of them are bedridden, which increases the chance of DVT and death from pulmonary embolism. Trauma increases the chance of having DVT. Low molecular weight heparin or heparin is a safe and extremely effective method of preventing DVT in high risk trauma patients. PMID- 16210989 TI - Robotics in colorectal surgery. AB - A minimally invasive approach has not yet become the gold standard in colorectal procedures, despite its proven advantages in postoperative recovery. This is in part the result of the technical limitations in today's standard laparoscopy, and the advanced surgical skills that are required. Robotic technology overcomes some of these limitations by successfully providing intuitive motion and enhanced precision and accuracy, in an environment that is much more ergonomic. While currently performed in only few designated centers, this technology has already been applied in almost every major procedure performed to treat both benign and malignant conditions of the large bowel. The feasibility of performing these procedures using robotic systems has been reported in several series. Conversion and complication rates are low, and short term results are comparable to conventional laparoscopy. However, no clear advantages to patients have been demonstrated yet. Furthermore, robotic technology is associated with a significant increase in time consumed during surgery and cost of care. Nevertheless, a great potential for patients benefit in the future may exist with this technology. Increasing clinical experience with these systems, further technological developments, and continuous research are required before robotic technology can be routinely incorporated into surgical procedures on the colon and rectum. PMID- 16210990 TI - Peritoneal dialysis catheter insertion. AB - Peritoneal dialysis (PD) catheter insertion can be accomplished by any 1 of 3 techniques. These include dissective or surgical, the blind or modified Seldinger, and laparoscopic techniques. The dissective technique solely utilized by surgeons, places the catheter by mini-laparotomy under general anesthesia. In the blind or modified Seldinger technique a needle is inserted into the abdomen, a guide-wire placed, a tract dilated and the catheter is inserted through a split sheath, all without visualization of the peritoneal cavity. Of the various laparoscopes, peritoneoscopic insertion uses a small optical peritoneoscope for direct inspection of the peritoneal cavity and identification of a suitable site for the intraperitoneal portion of the catheter. Hence, of the 3 techniques, only the latter allows for the direct visualization of the intraperitoneal structures. This technique can be easily used by nephrologists as well as surgeons. Peritoneoscopic placement varies from traditional laparoscopic techniques by using: a much smaller scope (2.2 mm diameter) and puncture size, only one peritoneal puncture site, a device to advance the cuff into the musculature, air in the peritoneum rather than CO2, and local anesthesia rather than general anesthesia. Prospective randomized and nonrandomized studies have shown that PD catheters peritoneoscopically placed by nephrologists have less incidence of complications (infection, exit site leak) and longer catheter survival rates than those inserted surgically. The current review focuses on the peritoneoscopic insertion of PD catheter and presents some of the complicating issues (bowel perforation, catheter migration, and prior abdominal surgery) related to this procedure. PMID- 16210991 TI - Preconditioning and the human heart. AB - Brief episodes of ischemia prior to coronary occlusion protect the heart during sustained coronary ischemia and is known as ischemic preconditioning. During acute myocardial infarction it is associated with smaller infarction size, less cardiac arrhythmias, and better left ventricular function. Brief balloon inflation in the cardiac catheterization laboratory during coronary intervention enables the operator to have further prolonged balloon inflations with lesser degrees of ischemia. Brief ischemia prior to coronary bypass surgery results in smaller perioperative infarctions and lesser degrees of postoperative arrhythmias. Preconditioning mimetic drugs may have a promising future in simulating ischemic preconditioning. PMID- 16210992 TI - Endothelium and inflammation. AB - During the past decade, our understanding of the pathophysiology of coronary heart disease (CAD) has undergone a remarkable evolution. To date atherosclerosis is considered an inflammatory disease, whose the endothelial dysfunction represents an early key event. When the arterial endothelium encounters certain bacterial products or risk factors, such as dyslipidemia, vasoconstrictor hormones involved in hypertension, the products of glycoxidation associated with hyperglycemia, or proinflammatory cytokines derived from excess adipose tissue, these cells increase the expression of adhesion molecules that promote the sticking of blood leukocytes to the inner surface of the arterial wall. Once in the arterial intima these cells communicate with endothelium and smooth muscle cells, under the influence of mediators of inflammation and immunity, such as the cytokines and complements components, prostanoids and leukotrienes. Thus, the activated endothelium promotes the development of the atherosclerotic disease process, i.e., vascular inflammation and thrombosis by producing vasoconstrictor substances, by inducing the expression of adhesive receptors for leukocytes and platelets, the production of tissue factor and endothelin, and by increasing the production of the plasminogen activator inhibitor-1. Emerging data support the concept that assessment of endothelial vasomotion may be a useful biomarker for atherosclerotic vascular disease. PMID- 16210993 TI - Inflammatory biomarkers of coronary atheromatous plaque vulnerability. AB - In the last decade, compelling evidence has evolved at both the basic science and clinical level for the implication of inflammation in the pathogenesis of atherosclerosis and its complications. The composition of the atherosclerotic plaque, rather than the degree of stenosis, is now recognized as a pivotal feature in determining plaque vulnerability and hence the risk of acute coronary ischaemic events. Current evidence supports a key role for inflammation in all phases of the atherosclerotic process, from plaque formation through to progression and, ultimately, the thrombotic complications of atherosclerosis. The growing appreciation of the role of inflammation in atherogenesis has focused attention on whether circulating levels of inflammatory biomarkers may help to identify those at risk of future cardiovascular events. In addition, the protective effects of a variety of interventions, such as statins, aspirin, and fibrates, are often associated with the evidence of reduced inflammation, further strengthening the notion that inflammation and the acute complications of atherosclerosis are causally related. The present review describes the pathophysiology of atheromatous plaque vulnerability and discusses the clinical use of inflammatory biomarkers for prognostic stratification of patients with acute coronary syndromes. PMID- 16210994 TI - Drug eluting stents in acute myocardial infarction. AB - Routine stent-implantation in primary coronary intervention (PCI) for acute myocardial infarction (AMI) has been shown to have a better clinical outcome than balloon angioplasty mainly because of reduction in restenosis rate and reocclusion. Drug eluting stents (DES) have recently been proven to further reduce restenosis and revascularization rate in comparison to bare metal stent (BMS) in elective procedures. Delayed endothelialization of these stents raises concern about a possible increase of thrombotic complications in the setting of AMI. Randomized studies with DES in the treatment of elective patients have shown at 9-12 months follow-up a thrombosis rate of 0-2% comparable to the one of BMS. Sirolimus eluting stents (SES) in AMI have been used in small series of consecutive pts not randomized or in registries with very high successful rate and a stent thrombosis varying between 0 and 4.7%. Paclitaxel eluting stent (PES) have also shown in small series a good immediate performance with a thrombosis rate between 0 and 4.8%. Predictors of acute and subacute stent thrombosis are the same than for BMS: residual dissection, long or overlapping stents, biforcation lesions and discontinuation of antiplatelets treatment. Providing effective mechanical reperfusion with similar results to the current therapeutic standard and decreasing the incidence of late complications, DES appear as an attractive approach for patients admitted with AMI. PMID- 16210995 TI - Minimally invasive (99m)Tc-sestamibi radioguided surgery of parathyroid adenomas. AB - In the last 10 to 15 years surgery of primary hyperparathyroidism (PHPT) moved from the wide bilateral neck exploration to various types of limited neck exploration ranging from unilateral neck surgery to minimally invasive approaches as the minimally invasive radioguided parathyroidectomy. In contrast with the bilateral neck exploration, an accurate preoperative localizing imaging, which is mainly based on (99m)TC-sestamibi scintigraphy, is mandatory when planning a concise parathyroidectomy. Following imaging criteria, only a fraction of PHPT patients accounting for approximately 60% to 70% of all PHPT patients can be eligible for a minimally invasive parathyroidectomy. Only PHPT patients with a high probability to be affected by a solitary parathyroid adenoma showing a high (99m)TC-sestamibi uptake and with a normal thyroid gland should be offer a minimally invasive radioguided parathyroidectomy. The (99m)TC-sestamibi SPECT technique and the double-tracer 123-iodine or (99m)TC-pertecnetate/(99m)TC sestamibi scintigraphic technique are the most sensitive and accurate preoperative imaging modalities and their utilization is recommended when considering a minimally invasive radioguided parathyroidectomy. Two main intraoperative procedures for the minimally invasive radioguided surgery have been described: the single-day protocol and the different-day protocol. In the single-day protocol a 740 MBq dose of (99m)TC-sestamibi is injected to the patient, scintigraphic imaging is obtained by dual-phase technique and then the patient is operated on within approximately 3 hours from radio-tracer injection. In the different-day protocol, a double-tracer parathyroid scintigraphy is obtained some days before surgery with the aim of better planning the type and extension of intervention. The day of intervention, for the purpose of radioguided surgery only, a low 37 MBq dose of (99m)TC-sestamibi is injected to the patient in the operating theatre a few minutes before the start of intervention. The main advantages of minimally invasive radioguided parathyroidectomy over the traditional wide bilateral neck exploration can be resumed as follows: a shortening in the operating and recovery time, possibility of local anesthesia, possibility of ambulatory surgery or same-day discharge, less postsurgical hypocalcemia, less postsurgical pain, favourable cosmetic results, benefits from a cost-analysis point of view. PMID- 16210996 TI - Current treatment options for asthma in adults. AB - Asthma is a common disorder with an increasing prevalence in the developed world. It is a serious health problem affecting health care costs, lost productivity, and death. Unfortunately, uncontrolled asthma is common and often unrecognized by physicians; of equal concern, uncontrolled asthma is accepted by asthmatic patients as they are uneducated as to what to expect from asthma therapy. Many learn to live with the limitations of daily activity and to overuse their rescue inhalers and yet when asked by their physicians, ''How's your asthma?'' the answer is ''okay''. The pursuit of more revealing questions regarding frequency in the use of rescue medications, nocturnal symptoms, activity limitations, and compliance with controller medications may be then overlooked. To achieve the goal of normal life on as few medications as possible will start with a well educated patient. Understanding intermittent from persistent asthma and then tailoring the best regimen for each patient is imperative. The treatment options and the advances in the understanding of the pathophysiology of asthma in the past 3 decades have been remarkable. There are increasing resources for both physicians and patients available to facilitate a better understanding of asthma management and the several treatment options. Currently established guidelines are an excellent starting point for initiating therapy for intermittent and mild, moderate, and severe persistent asthma. The short-acting beta 2 agonists are first-line therapy for intermittent asthma, exercise-induced asthma, and acute exacerbations. Long-acting Beta 2 agonists, cromolyn and nedocromil, and leukotriene modifiers may also be used for exercise. Systemic corticosteroids may be needed for acute exacerbations. The treatment of choice for persistent asthma is ICS and, depending on the severity, add-on therapy with long-acting beta 2 agonists, theophylline, and leukotriene modifiers are next. Lastly, omalizumab, anti-IgE therapy, will make an important place for its use in certain persistent moderate and severe asthma. This will be particularly true if the cost of the medication is reduced in the future. Asthma control, morbidity, and mortality will improve with a careful and comprehensive medical regimen using the current multiple treatment options. PMID- 16210997 TI - Hydrolyzed milk-serum peptides reduce body weight and fat content of dietary obese rats ameliorating their antioxidant status and liver functions. AB - AIM: Recent studies suggested that weight reduction under energy restriction required protein supplementation. Moreover, a significant decrease in serum cholesterol and triglycerides was observed when milk-serum proteins and, in particular, their hydrolyzed peptides were compared to milk casein. METHODS: Six Sprague-Dawley rats were fed with the standard diet for 8 weeks. Eighteen rats were fed with the obesity- producing diet for 4 weeks. After this period and for the remaining 4 weeks, these rats were divided into 3 groups, the 1st was fed with the obesity-diet, the 2nd and the 3rd were fed with the casein--and with the hydrolyzed milk-serum peptides--restricted diet, respectively. RESULTS: Treatment with the obesity-diet, compared to standard-diet, induced an increase in the body weight and fat content, with a decrease in protein mass and dehydration state. There was also an increase in blood levels of cholesterol, triglycerides and glucose. The lipoperoxides content in the plasma, heart, brain and liver had also increased, while the content of glutathione and ATP and the membrane fluidity in the liver had significantly decreased. The administration of the restricted caloric diet, in particular the one containing the hydrolyzed peptides were capable of an improvement of all these parameters. CONCLUSIONS: The metabolic modifications induced by the hydrolyzed peptides-restricted diet contribute to control better the over-weight thus reducing the risk of the onset of the dismetabolic pathologies correlated to it. PMID- 16210998 TI - The demonstration that I am sick is the fact that I have not recovered. PMID- 16210999 TI - Carcinogen metabolism, DNA damage repair and oral head and neck squamocellular carcinoma (HNSCC). A review. AB - Head and neck squamocellular carcinoma (HNSCC) has now become the 6th most common cancer among men in the developed world and affects the oral cavity, salivary glands, larynx and pharynx. Tobacco chewing, alcohol consumption and last but not least, smoking seem to be the most important risk factors. In particular in non drinkers, smoke increases the relative risk (RR) of developing HNSCC of the oral cavity and pharynx from 2 to 20 fold; especially in the oral cavity, the association between alcohol and smoke could have a multiplier effect. Cancer arises from damage to DNA of genes located at various points of the short (p) and long (q) arms of a number of chromosomes, caused by exposure to various carcinogens. Thus, the carcinogenic process requires continuous exposure to environmental carcinogens (i.e., longstanding history of smoking and drinking), an increased susceptibility to carcinogens (induced by xenobiotic metabolizing enzyme polymorphism) and an impaired DNA repair capacity (both inherited and acquired). Our purpose in this paper is to review advances in the understanding of the role of the European or Caucasian genetic aberrations that affect carcinogen metabolism and DNA repair genes in oral HNSCC development: we consider that those abnormalities will be useful in assessing individuals at risk. PMID- 16211000 TI - SEM qualitative evaluation of four self-etching adhesive systems. AB - AIM: Recently, there has been a tendency to simplify bonding procedures. Current self-etching adhesives combine conditioning, priming and bonding functions thus reducing technique-sensitivity as well as the risk of making errors. Another important advantage of this approach is that it allows the resin to etch and infiltrate the substratum simultaneously thus reducing the risk of any discrepancy between these 2 processes. The aim of this study was to evaluate the bonding quality of 4 self-etching adhesives both on dentin and enamel using SEM analysis. METHODS: Twenty caries-free molars were used. The aprismatic enamel layer was removed from the internal and external slopes of the vestibular cusps using a diamond bur. Each molar was then transversally cut just above the pulp chamber using a diamond saw (composhave 4255 Intensiv) thus obtaining twenty coronal fragments and 20 radicular fragments. Radicular and coronal fragments were then randomly divided into 5 groups. Each group consisted of 8 specimens: 4 coronal and 4 radicular ones. A different adhesive system was used for each group following the manufacturer's directions. Each group underwent the same treatment: a thin layer of flowable composite was applied and then polymerized. Subsequently, a single 2 mm layer of microhybrid composite was added. The coronal and radicular portions were longitudinally cut to show the ground enamel/adhesive, unground enamel/adhesive and dentine/adhesive interfaces. Eighty specimens were obtained and prepared for SEM observation. Forty specimens with an adhesive/enamel interface and 40 with an adhesive/dentine interface. RESULTS: The interfacial exam frequently showed gaps between the substratum and the restoration. The final etching was generally satisfactory on ground enamel, while it was poor on unground enamel. Sometimes gaps were evident at the adhesive interface in dentin specimens. The tested adhesive systems produced a suitable hybrid layer and a lot of resin-tags were present. CONCLUSIONS: Self-etching adhesives showed an adequate adhesion to dentin. However, while they provided encouraging results on ground enamel they did not show to be dependable on unground enamel. PMID- 16211001 TI - Magnetic resonance of the temporomandibular joint: experience at an Italian university center. AB - AIM: The aim of this investigation was to suggest criteria in order to evaluate magnetic resonance imaging (MRI) of the temporomandibular joint (TMJ). Such criteria have been discussed on the basis of our experience at the Section of Prosthetic Dentistry, Department of Neurosciences, University of Pisa. METHODS: The study sample was constituted by 135 patients. All subjects underwent bilateral MRI of the TMJs to evaluate disc structure and position, bony structure abnormalities, joint effusion localization and entity. RESULTS: MRI allowed depiction of the articular disc in 98.9% of the TMJs, showing a normal disc structure in 91.1% of the cases and abnormal in 7.7%. The disc-condyle relationship was normal in 46.6% TMJs, while a disc displacement with reduction was revealed in 35.5% cases, a disc displacement without reduction in 16.7% and a posterior disc displacement in 1.5% joints. In the coronal images, the disc was positioned lateral to the condyle in 8.9% of the TMJs and medial in 6.7%. Osseous abnormalities have been found in 177 joints (65.5%), with cases of bony flattening (condyle and/or tuberculum), erosions, subchondral cysts, osteophytosis and sclerosis. T2 sequences showed effusion in 26.7% of the TMJs. CONCLUSIONS: These findings suggest that standardized methodology application and well-defined criteria can facilitate MR imaging observations and interpretation as well as the diagnosis of intra-articular pathologies. PMID- 16211002 TI - Osteonecrosis of the jaws in patients treated with bisphosphonates. Review of the literature and the Milan experience. AB - AIM: A series of 5 patients with osteonecrosis of the jaws possibly related to the use of bisphosphonates is presented. A brief review of the literature is also proposed. METHODS: A retrospective review of patients who received bisphosphonates and who developed osteonecrosis of the jaws following minor oral surgery was performed. RESULTS: In all 5 patients, a tooth extraction preceded the onset of osteonecrosis of the mandible. Patients were on chronic use of intravenous bisphosphonates. The presenting lesions was a nonhealing extraction socket and an exposed jawbone. Biopsy of the bone lesions showed no evidence of metastatic involvement. Antibiotic therapy, conservative debridement and, in some cases, bone sequestrectomy were performed. CONCLUSIONS: Osteonecrosis of the jaws appears to have a relationship with the use of bisphosphonates. It is important that medical oncologists and dentists become aware of this possible complication in order to perform minor oral surgery prior to starting bisphosphonates treatment. PMID- 16211003 TI - Giant pleomorphic adenoma of the submandibular gland. Case report. AB - Salivary glands are sites for a great variety of tumors. The parotid gland is the most common site followed by the submandibular gland. The pleomorphic adenoma represents the 60-70% of all tumors involving the major salivary glands, while the submandibular gland is involved in 8% of cases. The pleomorphic adenoma of the submandibular gland is rare and can present difficulty in diagnosis. The heterogeneous histology, a possible malignant transformation, an incomplete capsule that can determine a recurrence, are the most important characteristics of pleomorphic adenoma. A complete surgical removal of the tumor is the treatment of choice. In the case presented, the mass showed an unusual vascularization and remarkable dimensions. PMID- 16211004 TI - Oculodentodigital dysplasia. A case report. AB - This report describes a rare case of oculodentodigital (ODD) dysplasia, an autosomic-dominant disease with alteration on the gene GJA1 of connexin 43 on the human chromosome 6q22-q23, highlighting the anomalies affecting face, eyes, teeth and limbs. The case described shows bilateral microphthalmia, microcornea, syndactyly and other phenotypic alterations characteristic of this syndrome, such as: nose of reduced size with hypoplasic wings, partial stenosis of the choanae, micrognathia, ogival palate and hypoplasia of the enamel. The authors emphasize the importance to know the principal features of ODD syndrome in order to make a correct diagnosis and the role of the dentist in the treatment of this pathology. PMID- 16211005 TI - Coronoid hyperplasia. A case report. AB - Coronoid hyperplasia is a rare condition which is macroscopically characterized by an increase in the dimensions of the coronoid process resulting from an abnormal bony elongation of histologically normal bone. Unilateral cases are more frequent than bilateral ones and can recognize a number of etiological factors, such as exostoses, osteochondroma, traumatic events, inflammatory reactions, neoplasia and manifestations secondary to other pathologies. Etiopathogenesis of bilateral forms has not yet been clarified: in the literature hints to both developmental and endocrine abnormalities are present, and a familiar pattern of inheritance has been evidenced. The poor specificity of signs and symptoms associated with coronoid hyperplasia, which are similar to those of other more frequent forms of temporomandibular disorders, present some problems of differential diagnosis. An accurate assessment must be based on a clinical and anamnestical approach aiming at the identification of pathognomonic clinical symptoms. Considering its limits (such as the possible presence of artefacts and image distortions), orthopantomography has a poor diagnostic usefulness. In this case, magnetic resonance (MR) allowed to evaluate disk-condyle relationship, but it could be also useful to evaluate post-operative complications. In the case of coronoid hyperplasia, the computed tomography (CT) is fundamental for a correct differential diagnosis. CT also allows surgical planning due to its accuracy to detect coronoid process volume and morphology. The present case report is an example of the need for a correct differential diagnosis between the different types of temporomandibular disorders, and it also lends support to the importance of requesting modern imaging techniques during the diagnostic process of the rare or complex cases. PMID- 16211006 TI - Loss of c-Cbl RING finger function results in high-intensity TCR signaling and thymic deletion. AB - Signaling from the T-cell receptor (TCR) in thymocytes is negatively regulated by the RING finger-type ubiquitin ligase c-Cbl. To further investigate this regulation, we generated mice with a loss-of-function mutation in the c-Cbl RING finger domain. These mice exhibit complete thymic deletion by young adulthood, which is not caused by a developmental block, lack of progenitors or peripheral T cell activation. Rather, this phenotype correlates with greatly increased expression of the CD5 and CD69 activation markers and increased sensitivity to anti-CD3-induced cell death. Thymic loss contrasts the normal fate of the c-Cbl-/ thymus, even though thymocytes from both mutant mice show equivalent enhancement in proximal TCR signaling, Erk activation and calcium mobilization. Remarkably, only the RING finger mutant thymocytes show prominent TCR-directed activation of Akt. We show that the mutant c-Cbl protein itself is essential for activating this pathway by recruiting the p85 regulatory subunit of PI 3-kinase. This study provides a unique model for analyzing high-intensity TCR signals that cause thymocyte deletion and highlights multiple roles of c-Cbl in regulating this process. PMID- 16211007 TI - Maturation of phage T7 involves structural modification of both shell and inner core components. AB - The double-stranded DNA bacteriophages are good model systems to understand basic biological processes such as the macromolecular interactions that take place during the virus assembly and maturation, or the behavior of molecular motors that function during the DNA packaging process. Using cryoelectron microscopy and single-particle methodology, we have determined the structures of two phage T7 assemblies produced during its morphogenetic process, the DNA-free prohead and the mature virion. The first structure reveals a complex assembly in the interior of the capsid, which involves the scaffolding, and the core complex, which plays an important role in DNA packaging and is located in one of the phage vertices. The reconstruction of the mature virion reveals important changes in the shell, now much larger and thinner, the disappearance of the scaffolding structure, and important rearrangements of the core complex, which now protrudes the shell and interacts with the tail. Some of these changes must originate by the pressure exerted by the DNA in the interior of the head. PMID- 16211008 TI - Distinct E2F-mediated transcriptional program regulates p14ARF gene expression. AB - The tumor suppressor p14(ARF) gene is induced by ectopically expressed E2F, a positive regulator of the cell cycle. The gene is expressed at low levels in normally growing cells in contrast to high levels in varieties of tumors. How p14(ARF) gene is regulated by E2F in normally growing cells and tumor cells remains obscure. Here we show that regulation of p14(ARF) gene by E2F is distinct from that of classical E2F targets. It is directly mediated by E2F through a novel E2F-responsive element that varies from the typical E2F site. The element responds to E2F activity resulting from ectopic E2F1 expression, inactivation of pRb by adenovirus E1a or shRNA, but not to phosphorylation of pRb by serum stimulation or ectopic cyclin D1/cyclin-dependent kinase-4 expression in normal human fibroblasts. The element has activity in various tumor cells with defective pRb, but not in normally growing cells. These results indicate that the distinct regulation constitutes the basis of p14(ARF) function as a tumor suppressor, discriminating abnormal growth signals caused by defects in pRb function from normal growth signals. PMID- 16211009 TI - KOPS: DNA motifs that control E. coli chromosome segregation by orienting the FtsK translocase. AB - Bacterial chromosomes are organized in replichores of opposite sequence polarity. This conserved feature suggests a role in chromosome dynamics. Indeed, sequence polarity controls resolution of chromosome dimers in Escherichia coli. Chromosome dimers form by homologous recombination between sister chromosomes. They are resolved by the combined action of two tyrosine recombinases, XerC and XerD, acting at a specific chromosomal site, dif, and a DNA translocase, FtsK, which is anchored at the division septum and sorts chromosomal DNA to daughter cells. Evidences suggest that DNA motifs oriented from the replication origin towards dif provide FtsK with the necessary information to faithfully distribute chromosomal DNA to either side of the septum, thereby bringing the dif sites together at the end of this process. However, the nature of the DNA motifs acting as FtsK orienting polar sequences (KOPS) was unknown. Using genetics, bioinformatics and biochemistry, we have identified a family of DNA motifs in the E. coli chromosome with KOPS activity. PMID- 16211010 TI - Structure and mechanisms of the proteasome-associated deubiquitinating enzyme USP14. AB - The ubiquitin-specific processing protease (UBP) family of deubiquitinating enzymes plays an essential role in numerous cellular processes. Mammalian USP14 (Ubp6 in yeast) is unique among known UBP enzymes in that it is activated catalytically upon specific association with the 26S proteasome. Here, we report the crystal structures of the 45-kDa catalytic domain of USP14 in isolation and in a complex with ubiquitin aldehyde, which reveal distinct structural features. In the absence of ubiquitin binding, the catalytic cleft leading to the active site of USP14 is blocked by two surface loops. Binding by ubiquitin induces a significant conformational change that translocates the two surface loops thereby allowing access of the ubiquitin C-terminus to the active site. These structural observations, in conjunction with biochemical characterization, identify important regulatory mechanisms for USP14. PMID- 16211014 TI - Newborn screening grows up. PMID- 16211016 TI - Research losses surface in hurricane Katrina's aftermath. PMID- 16211011 TI - Phosphorylation of ephrin-B1 via the interaction with claudin following cell-cell contact formation. AB - The interaction of the Eph family of receptor protein tyrosine kinase and its ligand ephrin family induces bidirectional signaling via the cell-cell contacts. Although most previous studies have focused on the function of Eph-ephrin pathways in the neural system and endothelial cells, this process also occurs in epithelial and cancer cells, of which the biological involvement is poorly understood. We show that ephrin-B1 creates an in vivo complex with adjacent claudin1 or claudin4 via the extracellular domains of these proteins. The cytoplasmic domain of ephrin-B1 was phosphorylated on tyrosine residues upon the formation of cell-cell contacts, possibly recognizing an intercellular adhesion of claudins. Phosphorylation of ephrin-B1 induced by claudins was abolished by the treatment with 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4 d]pyrimidine, an inhibitor of the Src family kinases. Moreover, overexpression of ephrin-B1 triggered consequent change in the level of cell-cell adhesion depending on its phosphorylation. These results suggest that ephrin-B1 mediated the cell-cell adhesion of epithelial and cancer cells via a novel Eph receptor independent mechanism. PMID- 16211017 TI - Women's health advocates dismayed at FDA's Plan B decision. PMID- 16211019 TI - Diabetes groups question whether 'metabolic syndrome' exists. PMID- 16211018 TI - Encephalitis outbreak finds Indian officials unprepared. PMID- 16211020 TI - Activists blame US policies for Uganda's condom shortage. PMID- 16211022 TI - Cancer researchers usher in dog days of medicine. PMID- 16211024 TI - Screen savers. PMID- 16211025 TI - Dominique Toran-Allerand. PMID- 16211026 TI - 60 years of winged victories for biomedical research. PMID- 16211027 TI - Perspectives on the properties of stem cells. PMID- 16211029 TI - Genetic fingerprinting. PMID- 16211028 TI - Tools for genomics. PMID- 16211030 TI - When society meets science. PMID- 16211032 TI - Old approach yields new antibiotic. PMID- 16211033 TI - Frog skin yields antiviral peptides. PMID- 16211034 TI - HIF, a missing link between metabolism and cancer. PMID- 16211035 TI - Fat, keeping the heart healthy? PMID- 16211036 TI - Fast track to the porphyrias. PMID- 16211037 TI - Hitchhiker's guide to the T cell. PMID- 16211038 TI - Targeting toxic proteins for turnover. PMID- 16211039 TI - Tweaking liver progenitor cells. PMID- 16211041 TI - The YIC factor. PMID- 16211042 TI - Korean biotechs seize opportunity to list on public markets. PMID- 16211043 TI - Chiron's manufacturing misfortunes boost competitors. PMID- 16211044 TI - New IGF drug stirs competition in growth factor segment. PMID- 16211045 TI - Big biotech embraces direct-to-consumer advertising. PMID- 16211047 TI - Profile: Jean Deleage. PMID- 16211048 TI - Biotech patenting slows in 2004. PMID- 16211049 TI - It came from beneath the sea. PMID- 16211052 TI - Decentralization key to ScanBalt. PMID- 16211050 TI - BIRB-796 is not an effective ABL(T315I) inhibitor. PMID- 16211053 TI - NCRI informatics initiative. PMID- 16211054 TI - A new role for scientists in the Biological Weapons Convention. PMID- 16211056 TI - A taxing dilemma. PMID- 16211055 TI - Mind the (biomedical funding) gap. PMID- 16211058 TI - A very firm handshake: biotech's growing negotiating power. PMID- 16211059 TI - ESTs stumble at the utility threshold. PMID- 16211060 TI - Recent patent applications in imaging technology. PMID- 16211061 TI - Islets for all? PMID- 16211063 TI - Simple recipe for blocking ion channels. PMID- 16211062 TI - Antibodies in the breakdown lane. PMID- 16211064 TI - Bacteria to the rescue. PMID- 16211065 TI - It takes a village to grow a tissue. PMID- 16211067 TI - Emergent behavior of growing knowledge about molecular interactions. PMID- 16211068 TI - Discovery of regulatory elements in vertebrates through comparative genomics. AB - We have analyzed issues of reliability in studies in which comparative genomic approaches have been applied to the discovery of regulatory elements at a genome wide level in vertebrates. We point out some potential problems with such studies, including difficulties in accurately identifying orthologous promoter regions. Many of these subtle analytical problems have become apparent only when studying the more complex vertebrate genomes. By determining motif reliability, we compared existing tools when applied to the discovery of vertebrate regulatory elements. We then used a statistical clustering method to produce a computational catalog of high quality putative regulatory elements from vertebrates, some of which are widely conserved among vertebrates and many of which are novel regulatory elements. The results provide a glimpse into the wealth of information that comparative genomics can yield and suggest the need for further improvement of genome-wide comparative computational techniques. PMID- 16211069 TI - Engineering novel binding proteins from nonimmunoglobulin domains. AB - Not all adaptive immune systems use the immunoglobulin fold as the basis for specific recognition molecules: sea lampreys, for example, have evolved an adaptive immune system that is based on leucine-rich repeat proteins. Additionally, many other proteins, not necessarily involved in adaptive immunity, mediate specific high-affinity interactions. Such alternatives to immunoglobulins represent attractive starting points for the design of novel binding molecules for research and clinical applications. Indeed, through progress and increased experience in library design and selection technologies, gained not least from working with synthetic antibody libraries, researchers have now exploited many of these novel scaffolds as tailor-made affinity reagents. Significant progress has been made not only in the basic science of generating specific binding molecules, but also in applications of the selected binders in laboratory procedures, proteomics, diagnostics and therapy. Challenges ahead include identifying applications where these novel proteins can not only be an alternative, but can enable approaches so far deemed technically impossible, and delineate those therapeutic applications commensurate with the molecular properties of the respective proteins. PMID- 16211075 TI - An individual approach. PMID- 16211082 TI - E75A and E75B have opposite effects on the apoptosis/development choice of the Drosophila egg chamber. AB - The number of Drosophila egg chambers is controlled by the nutritional status of the female. There is a developmental checkpoint at stage 8, which is controlled by BR-C in the follicle cells along with ecdysteroid. During this period, developmental decision is made in each egg chamber to determine if it will develop or die. During nutritional shortage, inducing apoptosis in the nurse cells of stages 8 and 9 egg chambers reduces the number of egg chambers. We show that ecdysone response genes E75A and E75B are involved in inducing or suppressing apoptosis. It is thus possible that the E75 isoforms A and B are involved in the decision to develop or die in oogenesis. We have established part of the pathway by which ecdysone response genes control apoptosis of the nurse cells and hence select between degeneration or development of individual egg chambers at stages 8 and 9. PMID- 16211083 TI - Murine Nod1 but not its human orthologue mediates innate immune detection of tracheal cytotoxin. AB - Tracheal cytotoxin (TCT) was originally described as the minimal effector that was able to reproduce the cytotoxic response of Bordetella pertussis on ciliated epithelial cells. This molecule triggers pleiotropic effects such as immune stimulation or slow-wave sleep modulation. Further characterization identified TCT as a specific diaminopimelic acid (DAP)-containing muropeptide, GlcNAc (anhydro)MurNAc-L-Ala-D-Glu-mesoDAP-D-Ala. Here, we show that the biological activity of TCT depends on Nod1, an intracellular sensor of bacterial peptidoglycan. However, Nod1-dependent detection of TCT was found to be host specific, as human Nod1 (hNod1) poorly detected TCT, whereas mouse Nod1 (mNod1) did so efficiently. More generally, hNod1 required a tripeptide (L-Ala-D-Glu mesoDAP) for efficient sensing of peptidoglycan, whereas mNod1 detected a tetrapeptide structure (L-Ala-D-Glu-mesoDAP-D-Ala). In murine macrophages, TCT stimulated cytokine secretion and NO production through Nod1. Finally, in vivo, injection of the tetrapeptide structure in mice triggered a transient yet strong release of cytokines into the bloodstream and the maturation of macrophages, in a Nod1-dependent manner. This study thereby identifies Nod1 as the long sought after sensor of TCT in mammals. PMID- 16211084 TI - A molecular switch and electronic circuit modulate catalase activity in catalase peroxidases. AB - The catalase reaction of catalase-peroxidases involves catalase-specific features built into a peroxidase core. An arginine, 20 A from the active-site heme, acts as a molecular switch moving between two conformations, one that activates heme oxidation and one that activates oxoferryl heme reduction by H(2)O(2), facilitating the catalatic pathway in a peroxidase. The influence of the arginine is imparted to the heme through its association with or dissociation from a tyrosinate that modulates reactivity through a Met-Tyr-Trp crosslinked adduct and a pi electron interaction of the heme with the adduct Trp. PMID- 16211085 TI - Bcr is a negative regulator of the Wnt signalling pathway. AB - The Wnt signalling pathway can activate transcription of genes such as c-myc through beta-catenin. Here, we describe the protein breakpoint cluster region, Bcr, as a negative regulator of this pathway. Bcr can form a complex with beta catenin and negatively regulate expression of c-Myc. Knockdown of Bcr by short interfering RNA relieves the block and activates expression of c-Myc. Expression of Bcr in the human colon carcinoma cell line HCT116, which has a high level of endogenous beta-catenin, leads to reduced c-Myc expression. The negative effect is exerted by the amino terminus of Bcr, which does not harbour the kinase domain. Bcr-Abl, the oncogene protein expressed in chronic myelogenous leukaemia (CML), does not bind to beta-catenin. It phosphorylates Bcr in the first exon sequence on tyrosines, which abrogates the binding of Bcr to beta-catenin. The inhibitor of the Bcr-Abl tyrosine kinase, STI-571 or Gleevec, a drug against CML, reverses this effect. Our data contribute to the understanding of Bcr as a tumour suppressor in the Wnt signalling pathway, as well as in CML. PMID- 16211086 TI - Gene therapy with drug resistance genes. AB - A major side effect of cancer chemotherapy is myelosuppression. Expression of drug-resistance genes in hematopoietic stem cells (HSC) using gene transfer methodologies holds the promise of overcoming marrow toxicity in cancer chemotherapy. Adequate protection of marrow cells in cancer patients from myelotoxicity in this way would permit the use of escalating doses of chemotherapy for eradicating residual disease. A second use of drug-resistance genes is for coexpression with a therapeutic gene in HSCs to provide a selection advantage to gene-modified cells. In this review, we discuss several drug resistance genes, which are well suited for in vivo selection as well as other newer candidate genes with potential for use in this manner. PMID- 16211087 TI - Transfection of Smac sensitizes tumor cells to etoposide-induced apoptosis and eradicates established human hepatoma in vivo. AB - A major concern in clinical treatment of cancers is resistance of tumors such as hepatocellular carcinoma (HCC) and osteosarcoma to current chemotherapy protocols. Here, we reported that overexpression of second mitochondria-derived activator of caspase (Smac) sensitized osteosarcoma cells and HCC cells in vitro to chemotherapeutic drugs-induced apoptosis. Constitutive expression of Smac resulted in enhanced Bax accumulation on mitochondria upon etoposide stimulation and inhibited Bcl-2-induced antiapoptosis activity. Thus, Smac would sensitize tumor cells to chemotherapeutic drugs in part through promoting Bax translocation to mitochondria and bypassing Bcl-2 block. Moreover, we demonstrated that blockade of Smac expression by antisense smac did not impair etoposide-induced apoptosis; however, p53-induced apoptosis was impaired in smac deficient Saos-2 cell. This suggested Smac might be required in p53-induced apoptosis. Most importantly, complete eradication of HepG2 xenografts in vivo was achieved upon combined therapy with Ad-Smac and 5-Fu. Thus, overexpression of Smac in tumor cells might be a potent strategy for cancer treatment by sensitization of tumor cells to chemotherapeutic drugs. PMID- 16211088 TI - Adenovirus-mediated p53 gene therapy in osteosarcoma cell lines: sensitization to cisplatin and doxorubicin. AB - The poor prognosis for patients with metastatic osteosarcoma (OS) indicates that new therapeutic options should be explored. Studies with adenoviral-mediated p53 gene transfer have been conducted in many cancer types including cervical, ovarian, prostatic and head and neck tumors. However, limited work has been carried out with pediatric cancers, including OS. Using three viral constructs containing cDNA for wild-type p53, mutant p53 (Cys135Ser) and lacZ, we studied the effect of adenoviral-mediated gene therapy in four OS cell lines: Saos-2 (p53 /-), HOS (R156P), KHOS/NP (R156P) and MNNG (R156P, F270L). We demonstrated that the virus efficiently enters the cells using the beta-galactosidase assay. Using the MTT assay, we have shown a dose-dependent decrease in cell viability 72 h post-treatment that occurs with Ad-wtp53 but not with Ad-mutp53. We have also shown that treatment with Ad-wtp53 significantly increases sensitivity of the cell lines to cisplatin and doxorubicin, chemotherapeutic agents commonly used in the treatment of OS. Our results indicate that restoration of wt p53 function in OS cells provides a basis for novel approaches to treatment of this disease. PMID- 16211089 TI - Overexpression of von Hippel-Lindau tumor suppressor protein and antisense HIF 1alpha eradicates gliomas. AB - The von Hippel-Lindau tumor suppressor protein (pVHL) suppresses tumor formation by binding the alpha subunits of hypoxia-inducible-factors responsible for stimulating tumor angiogenesis and glycolysis, and targeting them for ubiquitination and proteasomal destruction. Loss of pVHL leads to tumorigenesis and development of sporadic renal cell carcinomas and central nervous system hemangioblastomas. In the present study, we investigated whether engineered overexpression of pVHL in C6 glioma cells, which already express endogenous pVHL, would suppress the tumorigenicity of this particular tumor cell type. C6 cells overexpressing VHL displayed a reduced growth rate (70% inhibition) compared to the parental cell line when subcutaneously implanted in athymic (nu/nu) mice. Growth inhibition was associated with a 50% reduction in the number of tumor vessels and a 60% increase in tumor cell apoptosis, due in part to downregulation of HIF-1, VEGF, and the antiapoptotic factor Bcl-2, respectively. Gene transfer of VHL suppressed the growth of established C6 gliomas, and synergized with antisense HIF-1 to completely eradicate tumors. The data suggest that VHL gene therapy and/or agents that increase VHL expression could have utility in the treatment of gliomas, particularly when combined with agents that inhibit the expression or function of HIF-1. PMID- 16211090 TI - The role of 2 FOXP3 isoforms in the generation of human CD4+ Tregs. AB - Little is known about the molecules that control the development and function of CD4+ CD25+ Tregs. Recently, it was shown that the transcription factor FOXP3 is necessary and sufficient for the generation of CD4+ CD25+ Tregs in mice. We investigated the capacity of FOXP3 to drive the generation of suppressive CD4+ CD25+ Tregs in humans. Surprisingly, although ectopic expression of FOXP3 in human CD4+ T cells resulted in induction of hyporesponsiveness and suppression of IL-2 production, it did not lead to acquisition of significant suppressor activity in vitro. Similarly, ectopic expression of FOXP3delta2, an isoform found in human CD4+ CD25+ Tregs that lacks exon 2, also failed to induce the development of suppressor T cells. Moreover, when FOXP3 and FOXP3delta2 were simultaneously overexpressed, although the expression of several Treg-associated cell surface markers was significantly increased, only a modest suppressive activity was induced. These data indicate that in humans, overexpression of FOXP3 alone or together with FOXP3delta2 is not an effective method to generate potent suppressor T cells in vitro and suggest that factors in addition to FOXP3 are required during the process of activation and/or differentiation for the development of bona fide Tregs. PMID- 16211091 TI - Germinal center exclusion of autoreactive B cells is defective in human systemic lupus erythematosus. AB - Breach of B cell tolerance is central to the pathogenesis of systemic lupus erythematosus (SLE). However, how B cell tolerance is subverted in human SLE is poorly understood due to difficulties in identifying relevant autoreactive B cells and in obtaining lymphoid tissue. We have circumvented these limitations by using tonsil biopsies to study autoreactive B cells (9G4 B cells), whose regulation is abnormal in SLE. Here we show that 9G4 B cells are physiologically excluded during the early stages of the GC reaction before acquiring a centroblast phenotype. Furthermore, we provide evidence to indicate that an anergic response to B cell receptor stimulation may be responsible for such behavior. In contrast, in SLE, 9G4 B cells progressed unimpeded through this checkpoint, successfully participated in GC reactions, and expanded within the post-GC IgG memory and plasma cell compartments. The faulty regulation of 9G4 B cells was not shared by RA patients. To our knowledge, this work represents the first comparative analysis of the fate of a specific autoreactive human B cell population. The results identify a defective tolerance checkpoint that appears to be specific for human SLE. PMID- 16211092 TI - Pemphigus foliaceus IgG causes dissociation of desmoglein 1-containing junctions without blocking desmoglein 1 transinteraction. AB - Autoantibodies against the epidermal desmosomal cadherins desmoglein 1 (Dsg1) and Dsg3 have been shown to cause severe to lethal skin blistering clinically defined as pemphigus foliaceus (PF) and pemphigus vulgaris (PV). It is unknown whether antibody-induced dissociation of keratinocytes is caused by direct inhibition of Dsg1 transinteraction or by secondary cellular responses. Here we show in an in vitro system that IgGs purified from PF patient sera caused cellular dissociation of cultured human keratinocytes as well as significant release of Dsg1-coated microbeads attached to Dsg-containing sites on the keratinocyte cellular surface. However, cell dissociation and bead release induced by PF-IgGs was not caused by direct steric hindrance of Dsg1 transinteraction, as demonstrated by single molecule atomic force measurements and by laser trapping of surface-bound Dsg1 coated microbeads. Rather, our experiments strongly indicate that PF-IgG-mediated dissociation events must involve autoantibody-triggered cellular signaling pathways, resulting in destabilization of Dsg1-based adhesive sites and desmosomes. PMID- 16211093 TI - Modulation of atherosclerosis in mice by Toll-like receptor 2. AB - Epidemiologic evidence has established a relationship between microbial infection and atherosclerosis. Mammalian TLRs provide clues on the mechanism of this inflammatory cascade. TLR2 has a large ligand repertoire that includes bacterial derived exogenous and possibly host-derived endogenous ligands. In atherosclerosis-susceptible low-density lipoprotein receptor-deficient (Ldlr-/-) mice, complete deficiency of TLR2 led to a reduction in atherosclerosis. However, with BM transplantation, loss of TLR2 expression from BM-derived cells had no effect on disease progression. This suggested that an unknown endogenous TLR2 agonist influenced lesion progression by activating TLR2 in cells that were not of BM cell origin. Moreover, with intraperitoneal administration of a synthetic TLR2/TLR1 agonist, Pam3CSK4, disease burden was dramatically increased in Ldlr-/- mice. A complete deficiency of TLR2 in Ldlr-/- mice, as well as a deficiency of TLR2 only in BM-derived cells in Ldlr-/- mice, led to striking protection against Pam3CSK4-mediated atherosclerosis, suggesting a role for BM-derived cell expression of TLR2 in transducing the effects of an exogenous TLR2 agonist. These studies support the concept that chronic or recurrent microbial infections may contribute to atherosclerotic disease. Additionally, these data suggest the presence of host-derived endogenous TLR2 agonists. PMID- 16211095 TI - Synthetic mimics of mammalian cell surface receptors: prosthetic molecules that augment living cells. AB - Specific receptors on the surface of mammalian cells actively internalize cell impermeable ligands by receptor-mediated endocytosis. To mimic these internalizing receptors, my laboratory is studying artificial cell surface receptors that comprise N-alkyl derivatives of 3beta-cholesterylamine linked to motifs that bind cell-impermeable ligands. When added to living mammalian cells, these synthetic receptors insert into cellular plasma membranes, project ligand binding small molecules or peptides from the cell surface, and enable living cells to internalize targeted proteins and other cell-impermeable compounds. These artificial receptors mimic their natural counterparts by rapidly cycling between plasma membranes and intracellular endosomes, associating with proposed cholesterol and sphingolipid-rich lipid raft membrane microdomains, and delivering ligands to late endosomes/lysosomes. This "synthetic receptor targeting" strategy is briefly reviewed here and contrasted with other related cellular delivery systems. Potential applications of artificial cell surface receptors as molecular probes, agents for cellular targeting, tools for drug delivery, and methods for ligand depletion are discussed. The construction of synthetic receptors as prosthetic molecules, designed to seamlessly augment the molecular machinery of living cells, represents an exciting new frontier in the fields of bioorganic chemistry and chemical biology. PMID- 16211096 TI - A facile domino metathetic route to a thapsigargin skeleton. AB - A facile synthesis of a 5,7,5-fused ring system that is present in thapsigargins belonging to a novel family of sesquiterpene lactones, guainanolides, using domino enyne-RCM is reported here. PMID- 16211094 TI - A variant of SCID with specific immune responses and predominance of gamma delta T cells. AB - We describe here a patient with a clinical and molecular diagnosis of recombinase activating gene 1-deficient (RAG1-deficient) SCID, who produced specific antibodies despite minimal B cell numbers. Memory B cells were detected and antibodies were produced not only against some vaccines and infections, but also against autoantigens. The patient had severely reduced levels of oligoclonal T cells expressing the alphabeta TCR but surprisingly normal numbers of T cells expressing the gammadelta TCR. Analysis at a clonal level and TCR complementarity determining region-3 spectratyping for gammadelta T cells revealed a diversified oligoclonal repertoire with predominance of cells expressing a gamma4-delta3 TCR. Several gammadelta T cell clones displayed reactivity against CMV-infected cells. These observations are compatible with 2 non-mutually exclusive explanations for the gammadelta T cell predominance: a developmental advantage and infection triggered, antigen-driven peripheral expansion. The patient carried the homozygous hypomorphic R561H RAG1 mutation leading to reduced V(D)J recombination but lacked all clinical features characteristic of Omenn syndrome. This report describes a new phenotype of RAG deficiency and shows that the ability to form specific antibodies does not exclude the diagnosis of SCID. PMID- 16211097 TI - Membrane composition determines the fate of aggregated vesicles. AB - Vesicles incorporating a fluorescent metal-chelating lipid can be linked together by addition of copper(II) and poly-L-histidine, but the stability of adhering vesicles towards fusion depends upon membrane composition. PMID- 16211098 TI - Neighbouring group processes in the deamination of protonated phenylalanine derivatives. AB - The gas-phase fragmentation of protonated phenylalanine and a series of its derivatives (tyrosine, 4-methylphenylalanine, 4-aminophenylalanine, 4 methoxyphenylalanine, 4-tert-butylphenylalanine, 4-fluorophenylalanine, 4 chlorophenylalanine, 4-bromophenylalanine, 4-iodophenylalanine, 4 cyanophenylalanine, 4-nitrophenylalanine, 3-fluorophenylalanine, and 3,4 dichlorophenylalanine) were examined using a combination of low energy CID in a quadrupole ion trap mass spectrometer as well as DFT calculations and RRKM modelling. In particular, the relationship between the electron-donating ability of the substituent and the competitive losses of H2O + CO and NH3 were explored through the application of the Hammett equation. It was found that electron donating substituents promote the loss of NH3, while electron-withdrawing substituents suppress the loss of NH3 and favour the H2O + CO loss fragmentation channel instead. These observations are consistent with a neighbouring group pathway operating for the loss of NH3. Molecular orbital calculation (at the B3LYP/6-31+G(d,p) level of theory) were also performed for a range of derivatives to compare the relative transition state energy barriers for three competing mechanisms: (i) the combined loss of H2O + CO, which is triggered by an initial intramolecular proton transfer from the ammonium group to hydroxyl OH, followed by the combined loss of H2O and CO to form an immonium ion; (ii) loss of NH3 via an aryl assisted neighbouring group pathway to yield a phenonium ion; (iii) loss of NH3 via a 1,2-hydride migration process, which results in the formation of a benzyl cation. The relative energy barriers for H2O + CO loss remain nearly constant, while that for both NH3 pathways increase as the substituent moves from electron-donating to electron-withdrawing. The relative transition state energy for loss of NH3 via the aryl assisted neighbouring group pathway is always lower than that of the 1,2-hydride migration process. RRKM modelling of the DFT predicted barrier heights suggest that the rate constants for H2O + CO loss are insensitive to the substituent on the ring, while the NH3 loss channels are greatly affected by the substituent. These theoretical results are consistent with the experimental observation of the relative yields of the competing fragmentation channels. Finally, comparisons with published gas phase and condensed phase studies on related systems are made. PMID- 16211099 TI - Design and synthesis of aromatic inhibitors of anthranilate synthase. AB - Anthranilate synthase catalyses the conversion of chorismate to anthranilate, a key step in tryptophan biosynthesis. A series of 3-(1-carboxy-ethoxy) benzoic acids were synthesised as chorismate analogues, with varying functionality at C 4, the position of the departing hydroxyl group in chorismate. Most of the compounds were moderate inhibitors of anthranilate synthase, with inhibition constants between 20-30 microM. The exception was 3-(1-carboxy-ethoxy) benzoic acid, (C-4 = H), for which K(I)= 2.4 microM. These results suggest that a hydrogen bonding interaction with the active site general acid (Glu309) is less important than previously assumed for inhibition of the enzyme by these aromatic chorismate analogues. PMID- 16211100 TI - Total synthesis of milbemycins: a synthesis of (6R)-6-hydroxy-3,4 dihydromilbemycin E. AB - Following studies using benzyloxymethyl isopropenyl ketone 5 and ethyl 3-(3 furyl)-3-oxopropanoate 6, Robinson reactions between aryloxymethyl isopropenyl ketones 19 and 5 and ethyl 3-(2-trimethylsilyl-3-furyl)-3-oxopropanoate 20 were found to be stereoselective giving cyclohexanones 21 and 41, in which the 3 (arylmethoxy) substituents were cis to the 2-hydroxyl groups, as the major products. After reduction and protection of ketone 21, selective PMB deprotection, oxidation and stereoselective reduction inverted the configuration at C3 to give the diol 30. Protection of the secondary 3-hydroxyl group followed by modification of the protected 4-alcohol then gave the hydroxybutenolides 36 and 37 after oxidation of the silylated furan using singlet oxygen. The 3 benzyloxycyclohexanone 41 was also converted into the hydroxybutenolide 37 via the (2-trimethylsilylethoxy)methyl (SEM) ether 35. The Wittig reaction between the ylid generated from 2-methylpropyl(triphenyl)phosphonium salt and hydroxybutenolide 36 gave predominantly the (2Z,4Z)-dienyl acid 38 which was taken through to the butenolide 40. Similarly, the racemic hydroxybutenolide 37 was condensed with the racemic ylid derived from phosphonium salt 53 to give, after SEM-deprotection and 5-membered lactone formation, a mixture of the (9Z,2'Z)-dienyl lactones 58 and 59 containing ca. 10% of the corresponding (9Z,2'E)-isomers 60 and 61. (2'Z)/(2'E)-Isomerisation of the dienes 58 and 59 using iodine followed by deprotection gave a mixture of the seco-acids 62 and 63. Selective macrocyclisation of the seco-acid 62 in which the relative configuration of the C1-C7 and C17-C19 fragments (milbemycin numbering) corresponded to that present in the natural milbemycins, gave the beta-milbemycin analogue 65 after butenolide reduction. The hydroxybutenolide 37 was also condensed with the ylid derived from the phosphonium salt 1 and the product taken through to (6R)-6-hydroxy-3,4-dihydromilbemycin E 77. Preliminary attempts to convert the beta-milbemycin analogues 65 and 77 into tetrahydrofurans corresponding to analogues of alpha-milbemycins by treatment with toluene p sulfonyl chloride under basic conditions gave the primary allylic chlorides 78 and 79. PMID- 16211101 TI - A total synthesis of milbemycin G: approaches to the C1-C10-fragment and completion of the synthesis. AB - A synthesis of the hydroxybutenolide (-)-6 required for synthesis of alpha milbemycins and the completion of a total synthesis of milbemycin G 7 is reported. Following preliminary studies, an optimised synthesis of the hydroxybutenolide (-)-6 from the hydroxyketone 38 was developed which involved the resolution of 38 by separation of the 3-(O-chloroacetyl)-(S)-mandelates 80 and 83. Ester 80, which corresponded to the required enantiomer of the hydroxyketone 38, crystallized from the mixture of the diastereoisomeric esters 80 and 83 giving the (-)-hydroxyketone (-)-38 in an overall yield of 47%(based on racemic 38) after ethanolysis. Hydroxyketone (-)-38 was oxidised to the enolic diketone (-)-39 and phenylselenation and stereoselective reduction gave the trihydroxycyclohexyl selenide (-)-43. The regioselective introduction of the non conjugated double-bond into the six-membered ring was then achieved by esterification of the 4-hydroxyl group using trichloroacetic acid to give the trichloroacetate (-)-69. Oxidative elimination from the trichloroacetate using tert-butyl hydroperoxide was highly regioselective and gave the endo- and exocyclic alkenes (-)-44 and (-)-46 in a ratio of 95 : 5 after ethanolysis of the trichloroacetates. Selective O-methylation of the 4-hydroxyl group via the cyclic stannylene 55 and protection of the 3-hydroxyl group as its 2 trimethylsilylethoxymethyl (SEM) ether gave the ester (-)-57. Following saponification of the ethyl ester, re-esterification using 2 trimethylsilylethanol and oxidation of the 2-trimethylsilylfuryl fragment using singlet oxygen gave the required hydroxybutenolide (-)-6. The Wittig reaction between the phosphonium salt 2 and the hydroxybutenolide (-)-6 gave a ca. 2 : 1 mixture of the (4Z)- and (4E)-isomers of the ester 84 which on treatment with a catalytic amount of iodine was converted into the (4E)-isomer (4E)-84. Deprotection gave the seco-acid 85 but attempts to macrocyclise this were unsuccessful, the elimination product 86 being the only product isolated. The Wittig product 84 was taken through to the butenolide (2'E)-91 by removal of the SEM group, cyclisation to form the butenolide ring and diene isomerization, but this could not be converted into the corresponding seco-acid 92. However, removal of the SEM group from the seco-acid 85 gave the trihydroxy-acid 93 which was cyclized under modified Yamaguchi conditions to give the macrolide 94 together with a small amount of the macrocyclic butenolide 95. Reduction of this mixture using diisobutylaluminium hydride gave (6R)-6-hydroxymilbemycin E 96 which was converted to milbemycin G 7 by cyclisation of the primary chloride 97. The synthetic milbemycin G 7 was identical to a sample prepared by methylation of a commercial sample of milbemycin D 98, 7-O-methylmilbemycin G 99 being a side product of this methylation. PMID- 16211102 TI - Parallel synthesis and in vitro activity of novel anthranilic hydroxamate-based inhibitors of the prostaglandin H2 synthase peroxidase activity. AB - Currently available non-steroidal anti-inflammatory drugs (NSAIDs) such as aspirin are directed at the cyclooxygenase (COX) site, but not the peroxidase (POX) activity of prostaglandin H2 synthase (PGHS). They are thus unable to inhibit the free-radical induced tissue injury associated with PGHS peroxidase activity, which can occur independently of the COX site. A lead compound, anthranilic hydroxamic acid (AHA) was found to have significant PGHS-POX inhibitory activity (IC50= 72 microM). To define the critical parameters for PGHS POX inhibition, we investigated 29 AHA derivatives, synthesised from their acid precursors, using solid phase synthesis. In vitro analysis demonstrated a ten fold improvement in inhibition with 3,5-diiodoanthranilic hydroxamic acid (IC50= 7 microM). PMID- 16211103 TI - Baylis-Hillman reactions of N-tosyl aldimines and aryl aldehydes with 3 methylpenta-3,4-dien-2-one. AB - The attempted Baylis-Hillman reactions of N-tosyl aldimines and aryl aldehydes with 3-methylpenta-3,4-dien-2-one gave the corresponding Baylis-Hillman adducts 3 and 6 in moderate to good yields in the presence of DMAP in DMSO, respectively. In the case of the aza-Baylis-Hillman reactions of N-tosyl aldimines with 3 methylpenta-3,4-dien-2-one catalyzed by PBu3, the corresponding aza-Baylis Hillman derivatives 4 and 5 were formed at the same time. PMID- 16211104 TI - Micellar aggregates and hydrogels from phosphonobile salts. AB - The aggregation properties of novel bile acid analogs-phosphonobile salts (PBS) have been studied. The critical micellar concentration of 23 and 24-phosphonobile salts were measured using fluorescence and 31P NMR methods. All the ten synthesized phosphonobile salts formed gels at different pH ranges in water. The pH range at which individual PBSs could gelate water was narrow and influenced by the number and conformation of hydroxyl groups. A reversible thermochromic system has been developed (with 23-phosphonodeoxycholate at pH 3.3), which changes color upon gelation. The investigation of the first hydrogels derived from trihydroxy bile acid analogs 1 and 6 was made using fluorescence, 31P NMR, X-ray crystallography, circular dichroism and SEM. The present studies reveal that the gel network consists of a chiral, fibrous structure possessing hydrophobic interiors. PMID- 16211105 TI - Excited state tautomerization of azaindole. AB - Fluorescent tryptophan analogs, like azatryptophan, offer an advantage for exploring protein and peptide structure and dynamics. The chromophoric moieties, azaindole, of the azatryptophan analogs are investigated for their potential as fluorescent probes. The photophysical properties of 4-azaindole (4AI) and 5 azaindole (5AI) and their tautomers are characterized through computational and experimental methods. Both 4AI and 5AI undergo excited state tautomerization in the presence of 1 M NaOH. The protonated forms of 4AI and 5AI have a fluorescence emission of 415 and 410 nm, respectively, while the tautomers of 4AI and 5AI have a fluorescent emission of 480 and 450 nm, respectively. Gas phase computations (B3LYP/6-31+G**) show that the N1H azaindole tautomer is lower in energy in the ground state by as much as 12.5 kcal mol(-1), while the N(n)H azaindole tautomer is lower in energy in the excited state by as much as 18.1 kcal mol(-1). Solvent effects on the tautomer energy differences were computed using the isodensity polarized continuum model (IPCM). The polarity of the solvent helps to reduce the energy difference between the tautomers in the ground state by as much as 5.8 kcal mol(-1), but not enough to reverse the ground state tautomer preference. PMID- 16211106 TI - Analogs of the dihydroceramide desaturase inhibitor GT11 modified at the amide function: synthesis and biological activities. AB - Dihydroceramide desaturase is the last enzyme in the biosynthesis of ceramide de novo. The cyclopropene-containing sphingolipid GT11 is a competitive inhibitor of dihydroceramide desaturase. The biological effects of chemical modification of the GT11 amide linkage are reported in this article. Either N-methyl substitution or replacement of the amide alpha-carbonyl methylene by oxygen result in inactive compounds. In contrast, both urea (3) and thiourea (4) analogs of GT11, as well as three alpha-ketoamides (5-7), did inhibit the desaturation of N octanoylsphinganine to N-octanoylsphingosine, although with significantly lower potency than GT11. Furthermore, the alpha-ketoamides 5-7 inhibit the acidic ceramidase with similar potencies (IC50 52-83 microM). Inhibition of the neutral/alkaline ceramidase by these compounds requires around 20-fold higher concentrations. Structure-activity relationships and the biological interest of these compounds are discussed. PMID- 16211107 TI - Structure-activity relationships in 3-isothiazolones. AB - The biological activity of a series of structurally diverse 3-isothiazolones (1) has been assessed by evaluating the minimum inhibitory concentration required to inhibit the growth of E. Coli. The structure and electronic properties of these derivatives have been calculated using both semi-empirical and ab initio molecular orbital methods. Multi-linear regression analysis shows no correlation between the experimental activity of the 3-isothiazolones and either the calculated geometries, electronic properties, or the frontier orbital energies of these derivatives, but a reasonable relationship is found with other parameters including their calculated solvation energies, suggesting that diffusion may play an important role in their mode of action. PMID- 16211108 TI - Aziridine carboxylate from D-glucose: synthesis of polyhydroxylated piperidine, pyrrolidine alkaloids and study of their glycosidase inhibition. AB - The D-glucose derived aziridine carboxylate 5 was obtained from (E)-ethyl-6-bromo 1,2-O-isopropylidene-3-O-benzyl-5-deoxy-alpha-D-xylo-5-eno-heptofuranuronate 4 through conjugate addition of benzylamine and in situ intramolecular nucleophilic expulsion of bromine. The regioselective aziridine ring-opening, using water as a nucleophile, resulted in the alpha-hydroxy-beta-aminoester 6, which was exploited in the synthesis of six and five membered azasugars 1b/1c and 2b/2c, respectively. The glycosidase inhibitory activity of the title compounds was evaluated. PMID- 16211109 TI - Self-assembly and stability of double rosette nanostructures with biological functionalities. AB - The syntheses of calix[4]arene dimelamines that are functionalized with alkyl, aminoalkyl, ureido, pyridyl, carbohydrate, amino acid and peptide functionalities, and their self-assembly with barbituric acid or cyanuric acid derivatives into well-defined hydrogen-bonded nanostructures are described. The thermodynamic stability of these hydrogen-bonded assemblies was studied by CD spectroscopy in mixtures of CHCl3 and MeOH. The stability of the assemblies depends on several steric factors and the polarity of the functional groups connected to the assembly components. PMID- 16211110 TI - Chirality transfer in the aza-[2,3]-Wittig sigmatropic rearrangement. AB - The aza-[2,3]-Wittig sigmatropic rearrangements of substrates derived from enantiomerically pure alanine, valine and serine with phenyl and ester anion stabilising groups were investigated for their efficiency in chirality transfer. It was found that a methyl substituent at the stereogenic centre of the rearrangement precursors was inadequate to control the alkene stereoselectivity and enantioselectivity of the rearrangement. Ester stabilised anions of valine and serine derivatives were the most successful with up to 66% yield, 14 : 1 alkene (E)-stereoselection and 88% chirality transfer. A limitation to the steric bulk of the stereogenic centre was noted in that the substituent has to be bulky enough to dictate alkene stereoselection, but not too large to compromise the directing effect of the activating phenyldimethyl silyl substituent on the anion stabilising group. Experimental evidence suggested a possible complimentary coordinating effect of an O-MOM serine substituent, which may assist alkene stereoselectivity and enantioselectivity. PMID- 16211111 TI - Stereoselective beta-hydroxy-alpha-amino acid synthesis via an ether-directed, palladium-catalysed aza-Claisen rearrangement. AB - A highly diastereoselective synthesis of (2S,3S)-beta-hydroxy-alpha-amino acids has been developed from enantiopure alpha-hydroxy acids using a MOM-ether directed, palladium(II)-catalysed, aza-Claisen rearrangement of allylic acetimidates to effect the key step. This highly stereoselective process gave allylic amides in diastereomeric ratios of up to 14 : 1. Problems associated with the isolation of 1,3-products (anti-Claisen) from sterically demanding substrates via an insitu palladium(0)-catalysed rearrangement process were overcome by the addition of a re-oxidant, p-benzoquinone, leading to cleaner reactions and improved yields of the 3,3-products (Claisen). The target beta-hydroxy-alpha amino acids are an important class of natural products that are also components of more complex organic compounds with significant biological properties. PMID- 16211112 TI - Oligofunctional amphiphiles featuring geometric core group preorganization: synthesis and study of Langmuir and Langmuir-Blodgett films. AB - Based on the principle of supramolecular preorganization, a new type of oligofunctional amphiphile, of which compounds 1-4 are representative structures, has been designed and synthesized. The typical feature of their structure is a highly rigid and geometrically well-defined central unit composed of ethynylene substituted aromatic spacers with different numbers of amphiphilic segment groups (also of a rigid geometric design) attached to it. The molecules form well defined Langmuir films when spread from a solution at the air/water interface or when a 10(-4) M aqueous CaCl2 solution was used as the subphase. By analysis of the surface pressure-surface area (pi-A) isotherms, information on the packing behavior and orientation of the amphiphilic molecules depending on the molecular structure could be obtained. Morphological characterization of the dynamic process of monolayer compression at the air/water interface was carried out by Brewster angle microscopy, illustrating several phase states visualized as snap shots. Thin monolayer films produced on a 10(-4) M aqueous CaCl2 subphase can be transferred to a mica solid support by the Langmuir-Blodgett technique. Tapping mode atomic force microscopy reveals a surface topography of the monofilms composed of 1 and 3 that differ in roughness and also in the properties of elasticity, hardness and adhesive strength. X-Ray crystal structure analysis of three relevant intermediate compounds of the synthesis were successfully determined giving an indication of the potential structural features inherent in the new amphiphiles. PMID- 16211113 TI - Palladium catalysed aryl amination reactions in supercritical carbon dioxide. AB - Palladium catalysed C-N bond formation in supercritical carbon dioxide has been accomplished. Carbamic acid formation is avoided in part through the use of an N silylamine as the coupling partner. Employing a catalyst system of Pd2dba3(1 mol%) and 2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl (X-Phos)(2 mol%) enabled the catalytic amination of aryl bromides and chlorides with N silylanilines to be realised in excellent yield. Extension of the methodology to the N-arylation of N-silyldiarylamines, N-silylazoles and N-silylsulfonamides is reported. PMID- 16211114 TI - Synthesis of structures corresponding to the capsular polysaccharide of Neisseria meningitidis group A. AB - Four differently substituted trimers of the CPS repeating unit have been synthesised in order to investigate the dependence on oligosaccharide size, acetylation and mode of phosphorylation of glycoconjugate vaccines against Neisseria meningitidis group A. A spacer-containing starting monomer, a H phosphonate elongating monomer and a 6-O-phosphorylated H-phosphonate cap monomer have been synthesised and coupled together to afford, after deprotection, the target trimer structures differing in their acetylation and phosphorylation substitution pattern. PMID- 16211115 TI - From uncharged to decacationic molecules: syntheses and spectroscopic properties of heteroarenium-substituted pyridines. AB - Nucleophilic substitutions on pentachloropyridine with 4-(dimethylamino)pyridine, 4-aminopyridine, and 4-(pyrrolidin-1-yl)pyridine give mono-, tri- and pentacationic pyridine-hetarenium salts. The mono-, tri- and pentacationic 4 aminopyridine derivatives can be deprotonated to neutral compounds in solution, or protonated to di-, hexa- and decacationic pyridine derivatives, respectively. Successive substitutions with different heteroaromatic nucleophiles give pyridines with two distinct types of heteroarenium substituents. PMID- 16211116 TI - Rapid synthesis of quinoline-4-carboxylic acid derivatives from arylimines and 2 substituted acrylates or acrylamides under indium(III) chloride and microwave activations. Scope and limitations of the reaction. AB - Rapid synthesis of quinoline-4-carboxylic acid derivatives has been achieved by reaction of 2-methoxy acrylates or acrylamides with N-arylbenzaldimines in acetonitrile under InCl3 catalysis and microwave irradiation. Isolated yields up to 57% within 3 min have been obtained. The Lewis acid and the microwave activation appeared as crucial parameters for the reaction. The role of indium chloride and ytterbium triflate was specified using 13C NMR data and model theoretical studies. PMID- 16211117 TI - The diazo route to diazonamide A: studies on the tyrosine-derived fragment. AB - Various approaches to the tyrosine-derived fragment of the marine secondary metabolite diazonamide A are described. Initial efforts were focused on the originally proposed structure of the natural product, and a feasibility study established that a model 4-aryltryptamine could be readily prepared. Protected 4 bromotryptamine underwent Pd0-catalyzed coupling with the boronic acid derived from 2-bromophenyl allyl ether by Claisen rearrangement, O-methylation and lithiation-boration. The resulting biaryl was elaborated into an alpha-diazo-beta ketoester, dirhodium(II)-catalyzed reaction of which with N-Z-valinamide gave the desired tryptamine-oxazole following cyclodehydration of the intermediate ketoamide. A potential precursor to the benzofuran ring of the original structure of diazonamide A was prepared in eight steps from N-Z-tyrosine tert-butyl ester. Iodination, O-protection and Stille coupling gave the cinnamyl alcohol 25, converted via the bromide into the allyl aryl ether 27. Subsequent Claisen rearrangement and oxidative cleavage of the alkene gave the lactol 29, converted into the desired benzofuranone 31. The revision in the structure of diazonamide A to 2 resulted in the targeting of an alternative tyrosine-derived model benzofuranone 41 synthesized in four steps from N-Z-tyrosine methyl ester 36 by a route involving Claisen rearrangement of cinnamyl ether 37. Poor yields in this sequence prompted an investigation into the intramolecular Heck reaction as a route to benzofuranone 50. Coupling of 3-iodotyrosine 44 with 2-phenylbutenoic acid 48 gave ester 49 that readily underwent intramolecular Heck reaction to give benzofuranone 50, albeit with poor stereocontrol. PMID- 16211118 TI - Studies towards the synthesis of epothilone A via organoboranes. AB - Studies towards the synthesis of epothilone A via organoboranes have been described. A modified procedure for the large-scale preparation of B-gamma,gamma dimethylallyldiisopinocampheylborane from prenyl alcohol has been developed. This reagent, upon reaction with various aldehydes, provides the corresponding alpha,alpha-dimethylhomoallylic alcohols in high enantioselectivities. The application of this reagent for the synthesis of the C1-C6 subunit of epothilone has been demonstrated. Alternatively, inter- and intramolecular asymmetric reduction protocols have also been utilized for the synthesis of the C1-C6 subunit of epothilone A. The synthesis of the C7-C21 fragment of epothilone A involving asymmetric alkoxyallyl- and crotylboration using alpha-pinene-derived reagents has also been described. PMID- 16211119 TI - The prevalence of risk factors for coronary artery disease in patients who had had CABG. AB - The conclusion of Ranjith and his co-workers from their study 'Demographic data and the outcome of acute coronary syndrome in the South African Asian Indian population' drew our attention. Young South African Indians frequently have premature atherosclerosis with diffuse and aggressive disease. This should be taken into account for risk intervention. We would like to add to this. PMID- 16211120 TI - Cardiac markers--optimising proBNP in cardiac diagnosis and management. PMID- 16211122 TI - Dyslipidaemia among black patients with type 2 diabetes. AB - OBJECTIVE: To analyse the lipid profile of black patients with type 2 diabetes mellitus (type 2 DM) irrespective of duration or control of the disease. METHODS: This cross-sectional study evaluated the lipid profile of 62 black patients with type 2 DM. Clinical examination, fasting blood specimens for biochemistry, and urine analysis for proteinuria was determined in all patients. RESULTS: Only 25% of patients had adequately controlled blood pressure (BP < 140/90). Mean fasting plasma glucose (FPG) was 10.9 mmol/l (HbA1c: 10.6%). Mean body mass index (BMI) was 32.5. The total number of patients with diabetic dyslipidaemia (DD) was 56 (90.3%). Significant differences between men and women were noted for high density lipoprotein cholesterol (HDL-C) (0.99 vs 1.22 mmol/l, p = 0.005) and triglycerides (TG) (2.7 vs 1.8 mmol/l, p = 0.045), respectively. The commonest isolated abnormality was high LDL-C (66.1%) followed by low HDL-C (60.7%) and then high TG (57.1%). CONCLUSION: Over 90% of the black patients with type 2 DM displayed one or more features of DD affecting all major subclasses of cholesterol and triglyceride. The most frequent abnormality depicted by these patients was not the 'typical' DD (i.e. low HDL and high TG) often reported in other ethnic groups. PMID- 16211123 TI - Sustained white cell cytokine activation in idiopathic dilated cardiomyopathy despite haemodynamic improvement with medical therapy. AB - Cytokine production in idiopathic dilated cardiomyopathy (IDC) may depend on neurohumoral stimulation, haemodynamic impairment or auto-antibody production. We aimed to ascertain the impact of haemodynamic improvements with standard medical therapy and neurohumoral blockade on white cell tumour necrosis factor- alpha (TNF-alpha ) production in patients with IDC. Twenty-seven patients with IDC and NYHA class I to IV heart failure but without evidence of oedema, reduced peripheral perfusion, or elevated plasma endotoxin concentrations were evaluated for indicators of cytokine activation. Plasma TNF-alpha concentrations were raised (p < 0.001) in patients prior to commencement of medical therapy as compared to controls (n = 27). In addition, endotoxin-free cultured whole blood TNF-alpha production was enhanced (p < 0.02) in the patients. Although plasma TNF alpha tended to decrease, excessive whole blood TNF-alpha production remained unaltered following marked improvements in haemodynamics and functional class (increase in absolute left ventricular ejection fraction = 8.7 +/- 2.6%, p < 0.01, 37% in NYHA functional class I after therapy) with six to 12 months of medical therapy (diuretic, angiotensin converting enzyme inhibitor and beta blocker). Against a role for neurohumoral substances in promoting excessive white cell TNF-alpha synthesis the angiotensin II receptor antagonist, losartan, failed to modulate white cell TNF- alpha production in patients with IDC. We concluded that white cell TNF-alpha overproduction is sustained in patients with IDC despite haemodynamic improvement with standard medical therapy and blockade of angiotensin II receptors. These data suggest that mechanisms other than haemodynamic impairment and neurohumoral activation contribute to excess white cell TNF-alpha production in IDC. PMID- 16211124 TI - Cardiac findings in children admitted to a hospital general ward in South Africa: a comparison of HIV-infected and uninfected children. AB - The aim of this prospective observational study was to determine the presence of cardiac abnormalities in HIV infected versus uninfected children who were admitted to a general paediatric ward during a two-month period. HIV status was determined by antibody and p24 antigen testing. Clinical information, echocardiography and electrocardiography (ECG) were performed for all children. There were 90 HIV-infected and 118 uninfected children. The median age was 9.6 and 11.8 months for infected and uninfected children, respectively. Baseline left ventricular dysfunction, defined as a shortening fraction < or = 25%, was found in 13 (17%) of the HIV-infected children compared to 5 (8%) uninfected children (p < 0.05). Left ventricular end-diastolic enlargement above the 98th percentile for age was found in 24% of the infected and 20% of uninfected children. Pericardial effusions, although common, were sub-clinical and not different in the groups. ECG findings and resting heart rates were also similar. Left ventricular dysfunction was the most significant cardiac abnormality present in hospitalised HIV-infected children. Other abnormalities, although common, were mostly asymptomatic and found with the same frequency in uninfected children. Further studies are indicated. PMID- 16211125 TI - Transient ischaemic attack: is routine use of computerised cerebral tomography worthwhile? AB - AIM: To evaluate the role of routine computerised cerebral tomography (CCT) in patients undergoing carotid endarterectomy (CEA) following transient ischaemic attach (TIA). METHOD: A retrospective review was carried out of data collected on patients with TIA who were managed at the Durban Metropolitan Vascular Service over an 11-year period. All patients underwent carotid duplex scanning and CCT, followed by carotid endarterectomy (CEA) under general anaesthesia. RESULTS: The records of 140 patients were reviewed. There were 85 (61%) men and 55 (39%) women (F:M = 1:1.6). The mean age was 63 years (range 40-95 years). The presenting symptoms were amaurosis fugax, syncope and speech deficit. Risk factors for arterial disease were smoking (73%), hypertension (55%), diabetes mellitus (28%) and hyperlipidaemia (6%). Duplex ultrasonography showed significant internal carotid artery stenosis in 138 patients and occlusion in two. Arch angiography of the latter two patients showed pre-occlusive carotid stenosis. CCT scans showed abnormalities in 34 (24%) patients. These were lacunar infarcts (56%), old infarcts involving grey and white matter (32%), and cortical infarcts (12%). The findings did not influence management. All patients underwent routine CEA with selective shunting in 51 (36%) patients. Postoperative stroke rate was 2.9%. CONCLUSION: Routine CCT scanning in patients with TIA did not influence patient management and outcome. It is not cost effective and its routine use is questioned. PMID- 16211126 TI - The effect of compressed air massage on skin blood flow and temperature. AB - AIM: Compressed air massage is a new treatment modality that uses air under pressure to massage skin and muscle. It is claimed to improve skin blood flow but this has not been verified. Several pilot studies were undertaken to determine the effects of compressed air massage on skin blood flow and temperature. METHODS: Skin blood flow (SBF), measured using laser Doppler fluxmetry and skin temperature was recorded under several different situations: (i) treatment, at 1 Bar pressure using a single-hole (5-mm) applicator head, for 1 min at each of several sites on the right and left lower legs, with SBF measured on the dorsum of the left foot; (ii) at the same treatment pressure, SBF was measured over the left tibialis anterior when treatment was performed at different distances from the probe; (iii) SBF and skin temperature of the lower leg were measured with treatment at 0 or 1 Bar for 45 min, using two different applicator heads; (iv) SBF was measured on the dorsum of the foot of 10 subjects with treatment for 1 min at 0, 0.5, 1, 1.5 and 2 Bar using three different applicator heads. RESULTS: (i) SBF of the left foot was not altered by treatment of the right leg or chest, but was significantly increased during treatment of the left sole and first web, p < 0.0001. (ii) SBF over the tibialis anterior was increased when treatment was 5 cm from the probe, p < 0.0001, but not when 10 cm away. iii) SBF was significantly elevated throughout the 45-min treatments at 1 Bar and returned to normal within 1 min of stopping treatment. Skin temperature fell by 6.8 degrees C and 4.3 degrees C after 45-min treatments at 1 Bar, and slight rewarming occurred within 15 min. At 0 Bar, no change in SBF or temperature was noted. (iv) A near linear increase in SBF was noted with increasing treatment pressure for two of the three applicator heads. CONCLUSION: Compressed air massage causes an immediate increase in SBF, and an immediate fall in SBF when treatment is stopped. The effect appears to be locally and not centrally mediated and is related to the pressure used. Treatment cools the skin for at least 15 min after a 45-min treatment. PMID- 16211127 TI - Biventricular pacing for heart failure alters electro-mechanical coupling of both ventricles. AB - AIM: The exact mechanisms whereby biventricular pacing enables cardiac resynchronisation are not completely understood. This study looked at the effect of biventricular pacing on interventricular asynchrony in patients submitted to biventricular pacing. METHODS: A prospective series of 13 consecutive patients were selected from those referred for biventricular pacing. Criteria used included heart failure and QRS factors, as well as echocardiographic evidence of both intraventricular and interventricular asynchrony. Midterm follow-up of clinical and echocardiographic parameters are presented. RESULTS: All patients' clinical conditions improved significantly. Expectedly, diastolic filling parameters, ejection fraction and mitral regurgitation also improved significantly. The difference in the timing of left and right ventricular ejections of 65 ms at the baseline was corrected to 3 to 5 ms during six-month follow-ups after biventricular pacing. This effect was achieved by significant shortening of the left ventricular pre-ejection interval by 29 to 39 ms (p < 0.01) and by significant prolonging of the right ventricular pre-ejection interval by 18 to 30 ms (p < 0.01). CONCLUSION: Complete interventricular mechanical resynchronisation due to biventricular pacing occurred not only by the expected advancement in left ventricular ejection, but also by a delay in right ventricular ejection. The specific significance of correcting interventricular asynchrony with regard to the benefit and selection of patients for resynchronisation therapy remains to be fully established. PMID- 16211128 TI - Cardiovascular disease: hormone replacement therapy and the window of opportunity. PMID- 16211137 TI - Streptococcus suis infection. AB - A recent outbreak of Streptococcus suis infection associated with the slaughter, preparation or consumption of pigs in Sichuan, China has led to concerns that similar outbreaks could occur in other Asian countries. Although the pig farming industry is flourishing in Taiwan, reports of S. suis infection remain rare. We report 2 cases of S. suis meningitis successfully treated with ceftriaxone and penicillin. Previous reports of S. suis infection from the English literature are reviewed and the clinical data of cases reported in Asian and European countries are summarized. In Europe, there was good correlation between clinical disease and porcine contact, while few cases in Asia reported this association. Meningitis remained the most common presentation of infection in both areas (84.6% and 75.2%, respectively), followed by sepsis (15.4% and 18.6%, respectively), which had a higher mortality rate, particularly for splenectomized patients. Other clinical presentations included enteritis, arthritis, endocarditis, pneumonia, spondylodiscitis, endophthalmitis, uveitis and peritonitis. Deafness was a distinct sequelae (50.5% in Europe and 51.9% in Asia) after recovery from S. suis infection, especially in patients with meningitis. Not all commercial identification systems for streptococci could offer adequate speciation for S. suis. When viridans group streptococci are isolated from patients with meningitis and sepsis, prompt and correct identification of isolates to the species level should be performed, especially in areas with a high prevalence of S. suis diseases. PMID- 16211138 TI - Pediatric melioidosis in Pahang, Malaysia. AB - Melioidosis is much less common in children than in adults. This study investigated the incidence, demographic characteristics, presenting symptoms and outcome of pediatric melioidosis in Pahang, Malaysia. This retrospective study included patients < or =18 years old with positive body fluid cultures for Burkholderia pseudomallei from January 2000 to June 2003. Data on culture results were obtained from 2 referral hospitals. The incidence of pediatric melioidosis was 0.68/100,000 population per year. Of the 13 patients identified during the study period, 10 were male; 9 were Malays, 2 were Indians and 2 were aborigines. The mean age of these patients was 9.5 +/- 5.4 years. None of the patients had a previous history of confirmed melioidosis or predisposing factors for infection. Localized melioidosis was the most common presentation (46.2%) followed by melioidosis with septic shock (38.4%). Among patients with localized melioidosis, head and neck involvement (83.3%) was the most common presentation (2 patients with cervical abscesses, 1 with submandibular abscesses and 2 with acute suppurative parotitis) and another patient had right axillary abscess. All of the patients with septic shock had pneumonia and 2 of them had multi-organ involvement. The mortality among patients with septic shock was 80% and death occurred within 24 h of admission in all cases. In contrast, no complications or death occurred among patients with localized melioidosis. Melioidosis with septic shock is less common than localized melioidosis in pediatric patients, but is associated with very high mortality. PMID- 16211139 TI - Effect of melanin produced by a recombinant Escherichia coli on antibacterial activity of antibiotics. AB - A recombinant plasmid, pYL-1, containing a tyrosinase gene whose expression is under the control of a phage T5 promoter and 2 lac operators, was constructed. Escherichia coli JM109 harboring pYL-1 was used for production of bacterial melanin. A simple procedure for the isolation and purification of melanin was developed. The ultraviolet (UV)-visible light absorption spectra of melanin prepared by chemical synthesis and derived from different organisms, including bacteria, a plant and an animal source, were determined. Melanins produced by both bacteria and chemical synthesis showed a steady increase of absorption at wavelengths of UV light ranging from approximately 200-400 nm, while melanin derived either from plant or animal sources showed an additional discrete absorption peak at wavelength 280 nm upon a similar steady increase of absorption. This additional absorption peak could be due to the presence of protein-bound melanins in animal and plant sources while a free form of melanin was obtained from bacteria and chemical synthesis. Analysis of the effect of bacterial melanin on the activity of antibiotics against E. coli revealed that the activities of polymyxin B, kanamycin, tetracycline, and ampicillin were markedly reduced in the presence of melanin, whereas the activity of norfloxacin was not affected. The reduction of the antibacterial activity may result directly from the interaction of antibiotics with melanin. However, the mechanism of this interaction remains to be demonstrated. PMID- 16211140 TI - Comparison of the activities of granulocyte-macrophage colony-stimulating factor and interleukin-8 secretion between two lung epithelial cell lines. AB - The aim of this study was to survey the cytokine secretions in 2 human bronchial epithelial cell lines - a normal human bronchial epithelial cell line (HBEpC) and cell line A549, derived from malignant type II pneumocytes. The behavior of A549 cells is similar to epithelial cells and this line is widely used as an alternative model for studying human bronchial epithelial cell behavior. We measured the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-8 (IL-8) after tumor necrosis factor-alpha (TNF-alpha) or interleukin-1beta (IL-1beta) stimulation in the 2 cell lines. Both cell lines responded to TNF-alpha or IL-1beta stimulation, as shown by increased GM-CSF and IL-8 secretion. The relative cost, convenience and similarity of working with these 2 cell lines suggest that A549 is preferable for use as a first-line model and that results of studies of GM-CSF and/or IL-8 secretion under various stimulation conditions with this line could be confirmed using HBEpC. PMID- 16211141 TI - A neonatal echovirus 11 outbreak in an obstetric clinic. AB - An echovirus 11 outbreak occurred among neonates in an obstetric clinic in November 2003. Thirteen neonates were transferred to our medical center, and all were found to have echovirus 11 infection. Viral studies were performed for 32 other infants born in the clinic during the same period, including 30 asymptomatic neonates and 2 febrile infants transferred to another hospital. Two of the asymptomatic infants had echovirus 11 isolated from rectal swabs. The first patient transferred to our medical center developed extensive hemorrhage and died 6 days later. Three family members of this infant were also proved to have echovirus 11 infections. One other infant had a fulminant course and had residual hepatic impairment. The other infants had no complications. Viral studies in the 24 nursery staff were all negative. This outbreak shows how a neonatal enterovirus outbreak can occur in a nursery, starting from an infected infant in the incubation period. Early recognition and prompt management of an outbreak is important to prevent further spread of the infection. PMID- 16211142 TI - Food-specific immunoglobulin E among children with atopic dermatitis: a retrospective study. AB - This retrospective study included 133 children aged between 2 and 16 years with elevated serum food-specific immunoglobulin E (IgE), including 59 children clinically diagnosed with atopic dermatitis (AD) and 74 children clinically diagnosed with atopic disease without AD (asthma, allergic rhinitis, or both). Six common serum food-specific IgEs were detected by the Pharmacia ImmunoCAP test, including: egg white, milk, peanut, soybean, shrimp and egg yolk. Serum total IgE was also measured. The results showed that both AD and non-AD atopic children had the highest sensitization rate to shrimp. AD children had significantly higher serum total IgE and average number of positive food sensitization items than atopic children without AD. Three serum food-specific IgEs, including peanut, soybean and egg yolk, were significantly higher in children with AD than in those without AD. Furthermore, 3 pairs of food-specific IgEs were correlated with each other in AD children: egg white IgE correlated with peanut IgE, egg white IgE correlated with egg yolk IgE, and peanut IgE correlated with soybean IgE. In logistic regression analysis of the serum of 6 food allergen-specific IgEs in AD children, we found that elevated peanut- and egg yolk-specific IgE were risk factors of AD in elevated serum food-specific IgE children whose serum total IgE was less than 1000 kU/L but not in those with total IgE greater than 1000 kU/L. PMID- 16211143 TI - A retrospective study of hantavirus infection in Kinmen, Taiwan. AB - In this retrospective study, 140 serum samples collected from 85 scrub typhus negative patients in Kinmen Island in 2000 were tested for antibodies to hantavirus using enzyme-linked immunosorbent assay. Seven patients (8.23%) were confirmed as having hantavirus infection as demonstrated by increased hantavirus specific immunoglobulin M and/or immunoglobulin G antibodies in their convalescent serum samples. Analysis of indirect immunofluorescence assay showed that Seoul type was the etiologic agent. Serosurvey of rodents caught in the resident township of these hantavirus-infected human cases showed that the seroprevalence of antibodies to hantavirus among Rattus norvegicus, Mus musculus, and R. flavipectus was 50% (4/8), 20% (1/5), and 2% (7/348), respectively. Molecular analysis showed that these reservoir hosts carried a Seoul type hantavirus. To our knowledge, this is the first report demonstrating indigenous hantavirus cases in Kinmen. PMID- 16211144 TI - Serratia marcescens bacteremia at a medical center in southern Taiwan: high prevalence of cefotaxime resistance. AB - Antimicrobial resistance of isolates and risk factors for mortality were retrospectively investigated in 71 adult patients with Serratia marcescens bacteremia. During the 4-year study period, 78 clinically significant episodes of S. marcescens bacteremia occurred in 71 patients. The mean age of the patients was 65 years (range, 25-86 years) with a male predominance (45 patients, 63%). Most of the bacteremic episodes were nosocomial (78%), and 34% were polymicrobial. The overall mortality rate within 2 weeks after the onset of bacteremia was 41%. The presence of malignancy and critical illness at initial presentation were independent risk factors for mortality. By disk susceptibility test, 72 isolates were resistant to cefotaxime (92%) but susceptible to ceftazidime (99%). All isolates were susceptible to meropenem. Among the 47 patients with monomicrobial S. marcescens bacteremia, the mortality rate within 5 days of onset in patients receiving appropriate empirical antimicrobial therapy was lower than that in patients receiving inappropriate therapy although this difference was not significant (14% vs 28%, p = 0.27). Among the patients with cefotaxime-resistant but ceftazidime-susceptible S. marcescens bacteremia treated with ceftazidime, 6 of 7 patients (86%) survived for more than 2 weeks, suggesting the potential effectiveness of ceftazidime in the treatment of cefotaxime-resistant Serratia infections. Further clinical studies are required to delineate the clinical role of ceftazidime therapy for infections caused by S. marcescens with this resistant phenotype. PMID- 16211145 TI - Common variable immunodeficiency mimicking rheumatoid arthritis with Sjogren's syndrome. AB - Several autoimmune diseases have been reported to be associated with common variable immunodeficiency disease (CVID), including rheumatoid arthritis and Sjogren's syndrome. On the other hand, approximately 20-30% of patients with rheumatoid arthritis develop secondary Sjogren's syndrome. A 26-year-old woman had a 6-year history of chronic symmetric polyarthritis and 3-year history of sicca syndrome prior to admission for pneumonia. Rheumatoid arthritis with secondary Sjogren's syndrome had been diagnosed 1 year before. The patient had experienced 3 episodes of pneumonia during the previous 3 years. Markedly depressed serum immunoglobulin levels prompted a suspicion of common variable immunodeficiency, and the impression was confirmed after a series of examinations. Monthly administration of intravenous immunoglobulin (IVIG) alleviated the polyarthritis and improved the sicca syndrome. IVIG replacement therapy was ultimately successful in curing recurrent bacterial infections, chronic polyarthritis, and improving the severity of sicca syndrome. PMID- 16211146 TI - Rapidly progressive necrotizing fasciitis caused by Staphylococcus aureus. AB - Necrotizing fasciitis (NF) is a rapidly progressive life-threatening infection located in the deep fascia, with secondary necrosis of the subcutaneous tissues. Staphylococcus aureus as a single etiologic agent is rare. The pathogenicity of S. aureus infections is related to various bacterial surface components and extracellular proteins. A 56-year-old man developed fever, hypotension, impaired renal and hepatic functions, disseminated intravascular coagulation, and rapidly progressive NF affecting the 4 extremities due to methicillin-susceptible S. aureus (MSSA). The initial presenting symptoms were general weakness and muscular pain over bilateral thighs and left shoulder, and gradual onset of weakness of the limbs. On the third hospital day, multiple red-purplish discoloration spread across the right lower leg and left forearm. Fasciotomy and debridement was performed on the fifth hospital day, and the diagnosis of NF was confirmed. MSSA was the only pathogen isolated from 4 sets of blood cultures taken on admission and cultures of tissues collected during surgical debridement. The disease progressed rapidly over the 4 extremities despite appropriate antibiotic treatment. He recovered after multiple extensive surgical interventions and 8 weeks of intensive medical care. Early diagnosis, intensive surgical intervention, antibiotic treatment and intensive medical care are crucial for a successful outcome in patients with septic shock and extensive NF caused by S. aureus. PMID- 16211147 TI - Successful preventive treatment of congenital heart block during pregnancy in a woman with systemic lupus erythematosus and anti-Sjogren's syndrome A/Ro antibody. AB - Congenital heart block (CHB) that is a manifestation of neonatal lupus syndrome (NLS) carries a poor prognosis. The treatment response of established heart block in NLS is usually unsatisfactory. Preventive treatment during pregnancy, however, before the critical period of cardiac development, can prevent the development of CHB. A Taiwanese woman with systemic lupus erythematosus (SLE) was positive for anti-Sjogren's syndrome A (SSA)/Ro antibody. Her first pregnancy resulted in intra-uterine fetal death. Her second pregnancy resulted in CHB, despite dexamethasone treatment, and neonatal death at age 1 day despite pacemaker implantation. During her third pregnancy, dexamethasone was given starting at week 10, azathioprine at week 18, and plasmapheresis was performed every other day for 5 times starting at week 20 of gestation. Cesarean section was performed due to oligohydramnion at week 31 of gestation and a healthy girl was delivered. This case suggests that judicious use of fluorinated glucocorticoids, immunosuppressants, and plasmapheresis may prevent development of CHB in pregnant women with SLE who are anti-SSA/Ro antibody positive and have previous children with CHB. PMID- 16211149 TI - WHO's action for continuous improvement in oral health. PMID- 16211150 TI - Tobacco or oral health. PMID- 16211151 TI - Oral health, general health and quality of life. PMID- 16211153 TI - More oral health care needed for ageing populations. PMID- 16211155 TI - Reasons for attending dental-care services in Ouagadougou, Burkina Faso. AB - OBJECTIVE: To determine why patients attend dental-care facilities in Ouagadougou, Burkina Faso and to improve understanding of the capacity of oral health-care services in urban west Africa. METHODS: We studied a randomly selected sample of patients attending 15 dental-care facilities in Ouagadougou over a 1-year period in 2004. Data were collected using a simple daily record form. FINDINGS: From a total of 44,975 patients, the final sample was established at 14,591 patients, of whom 55.4% were new patients and 44.6% were "booking patients". Most patients seeking care (71.9%) were aged 15-44 years. Nongovernmental not-for-profit dental services were used by 41.5% of all patients, 36% attended private dental-care services, and 22.5% of patients visited public services. The most common complaint causing the patient to seek dental-care services was caries with pulpal involvement (52.4%), and 60% of all complaints were associated with pain. The patients' dental-care requirements were found to differ significantly according to sex, health insurance coverage and occupation. CONCLUSION: Urban district health authorities should ensure provision of primary health-care services, at the patients' first point of contact, which are directed towards the relief of pain. In addition to the strengthening of outreach emergency care, health centres should also contribute to the implementation of community-based programmes for the prevention of oral disease and the promotion of oral health. Exchange of experiences from alternative oral health-care systems relevant to developing countries is urgently needed for tackling the growing burden of oral disease. PMID- 16211156 TI - Areca (betel) nut chewing habit among high-school children in the Commonwealth of the Northern Mariana Islands (Micronesia). AB - OBJECTIVE: To investigate the prevalence of its use by high-school children in Saipan in Micronesia. Usage of the areca nut is indigenous to south Asia and the western and south Pacific. Some serious health effects of areca nut chewing are recognized and the International Agency for Research on Cancer has recently classified regular use of areca nut as being carcinogenic to humans. Information on usage by young people, however, is scarce. METHODS: Data on consumption of areca nut were obtained by a self-administered questionnaire. Following an oral mucosal examination using WHO criteria any detectable oral mucosal diseases were recorded. FINDINGS: Of 309 schoolchildren surveyed (mean age 16.3 +/- 1.5 years), 63.4% claimed regular use, the highest level recorded in any school population survey. Significant oral diseases detected were oral leukoplakia in 13% and oral submucous fibrosis in 8.8% of children. CONCLUSION: These findings from Saipan suggest that areca nut chewing starts at a young age in Micronesia. As many users develop dependency this raises important concerns regarding its consequences for oral health. PMID- 16211157 TI - The global burden of oral diseases and risks to oral health. AB - This paper outlines the burden of oral diseases worldwide and describes the influence of major sociobehavioural risk factors in oral health. Despite great improvements in the oral health of populations in several countries, global problems still persist. The burden of oral disease is particularly high for the disadvantaged and poor population groups in both developing and developed countries. Oral diseases such as dental caries, periodontal disease, tooth loss, oral mucosal lesions and oropharyngeal cancers, human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS)-related oral disease and orodental trauma are major public health problems worldwide and poor oral health has a profound effect on general health and quality of life. The diversity in oral disease patterns and development trends across countries and regions reflects distinct risk profiles and the establishment of preventive oral health care programmes. The important role of sociobehavioural and environmental factors in oral health and disease has been shown in a large number of socioepidemiological surveys. In addition to poor living conditions, the major risk factors relate to unhealthy lifestyles (i.e. poor diet, nutrition and oral hygiene and use of tobacco and alcohol), and limited availability and accessibility of oral health services. Several oral diseases are linked to noncommunicable chronic diseases primarily because of common risk factors. Moreover, general diseases often have oral manifestations (e.g. diabetes or HIV/AIDS). Worldwide strengthening of public health programmes through the implementation of effective measures for the prevention of oral disease and promotion of oral health is urgently needed. The challenges of improving oral health are particularly great in developing countries. PMID- 16211158 TI - The effective use of fluorides in public health. AB - Dental caries remain a public health problem for many developing countries and for underprivileged populations in developed countries. This paper outlines the historical development of public health approaches to the use of fluoride and comments on their effectiveness. Early research and development was concerned with waterborne fluorides, both naturally occurring and added, and their effects on the prevalence and incidence of dental caries and dental fluorosis. In the latter half of the 20th century, the focus of research was on fluoride toothpastes and mouth rinses. More recently, systematic reviews summarizing these extensive databases have indicated that water fluoridation and fluoride toothpastes both substantially reduce the prevalence and incidence of dental caries. We present four case studies that illustrate the use of fluoride in modern public health practice, focusing on: recent water fluoridation schemes in California, USA; salt fluoridation in Jamaica; milk fluoridation in Chile; and the development of "affordable" fluoride toothpastes in Indonesia. Common themes are the concern to reduce demands for compliance with fluoride regimes that rely upon action by individuals and their families, and the issue of cost. We recommend that a community should use no more than one systemic fluoride (i.e. water or salt or milk fluoridation) combined with the use of fluoride toothpastes, and that the prevalence of dental fluorosis should be monitored in order to detect increases in or higher-than-acceptable levels. PMID- 16211159 TI - Health-promoting schools: an opportunity for oral health promotion. AB - Schools provide an important setting for promoting health, as they reach over 1 billion children worldwide and, through them, the school staff, families and the community as a whole. Health promotion messages can be reinforced throughout the most influential stages of children's lives, enabling them to develop lifelong sustainable attitudes and skills. Poor oral health can have a detrimental effect on children's quality of life, their performance at school and their success in later life. This paper examines the global need for promoting oral health through schools. The WHO Global School Health Initiative and the potential for setting up oral health programmes in schools using the health-promoting school framework are discussed. The challenges faced in promoting oral health in schools in both developed and developing countries are highlighted. The importance of using a validated framework and appropriate methodologies for the evaluation of school oral health projects is emphasized. PMID- 16211160 TI - Oral health information systems--towards measuring progress in oral health promotion and disease prevention. AB - This article describes the essential components of oral health information systems for the analysis of trends in oral disease and the evaluation of oral health programmes at the country, regional and global levels. Standard methodology for the collection of epidemiological data on oral health has been designed by WHO and used by countries worldwide for the surveillance of oral disease and health. Global, regional and national oral health databanks have highlighted the changing patterns of oral disease which primarily reflect changing risk profiles and the implementation of oral health programmes oriented towards disease prevention and health promotion. The WHO Oral Health Country/Area Profile Programme (CAPP) provides data on oral health from countries, as well as programme experiences and ideas targeted to oral health professionals, policy makers, health planners, researchers and the general public. WHO has developed global and regional oral health databanks for surveillance, and international projects have designed oral health indicators for use in oral health information systems for assessing the quality of oral health care and surveillance systems. Modern oral health information systems are being developed within the framework of the WHO STEPwise approach to surveillance of noncommunicable, chronic disease, and data stored in the WHO Global InfoBase may allow advanced health systems research. Sound knowledge about progress made in prevention of oral and chronic disease and in health promotion may assist countries to implement effective public health programmes to the benefit of the poor and disadvantaged population groups worldwide. PMID- 16211161 TI - The role of diet and nutrition in the etiology and prevention of oral diseases. AB - Diet plays an important role in preventing oral diseases including dental caries, dental erosion, developmental defects, oral mucosal diseases and, to a lesser extent, periodontal disease. This paper is intended to provide an overview of the evidence for an association between diet, nutrition and oral diseases and to clarify areas of uncertainty. Undernutrition increases the severity of oral mucosal and periodontal diseases and is a contributing factor to life-threatening noma. Undernutrition is associated with developmental defects of the enamel which increase susceptibility to dental caries. Dental erosion is perceived to be increasing. Evidence suggests that soft drinks, a major source of acids in the diet in developed countries, are a significant causative factor. Convincing evidence from experimental, animal, human observational and human intervention studies shows that sugars are the main dietary factor associated with dental caries. Despite the indisputable role of fluoride in the prevention of caries, it has not eliminated dental caries and many communities are not exposed to optimal quantities of fluoride. Controlling the intake of sugars therefore remains important for caries prevention. Research has consistently shown that when the intake of free sugars is < 15 kg/person/year, the level of dental caries is low. Despite experimental and animal studies suggesting that some starch-containing foods and fruits are cariogenic, this is not supported by epidemiological data, which show that high intakes of starchy staple foods, fruits and vegetables are associated with low levels of dental caries. Following global recommendations that encourage a diet high in starchy staple foods, fruit and vegetables and low in free sugars and fat will protect both oral and general health. PMID- 16211162 TI - Oral lesions in infection with human immunodeficiency virus. AB - This paper discusses the importance of oral lesions as indicators of infection with human immunodeficiency virus (HIV) and as predictors of progression of HIV disease to acquired immunodeficiency syndrome (AIDS). Oral manifestations are among the earliest and most important indicators of infection with HIV. Seven cardinal lesions, oral candidiasis, hairy leukoplakia, Kaposi sarcoma, linear gingival erythema, necrotizing ulcerative gingivitis, necrotizing ulcerative periodontitis and non-Hodgkin lymphoma, which are strongly associated with HIV infection, have been identified and internationally calibrated, and are seen in both developed and developing countries. They may provide a strong indication of HIV infection and be present in the majority of HIV-infected people. Antiretroviral therapy may affect the prevalence of HIV-related lesions. The presence of oral lesions can have a significant impact on health-related quality of life. Oral health is strongly associated with physical and mental health and there are significant increases in oral health needs in people with HIV infection, especially in children, and in adults particularly in relation to periodontal diseases. International collaboration is needed to ensure that oral aspects of HIV disease are taken into account in medical programmes and to integrate oral health care with the general care of the patient. It is important that all health care workers receive education and training on the relevance of oral health needs and the use of oral lesions as surrogate markers in HIV infection. PMID- 16211163 TI - Managing severe acute respiratory syndrome (SARS) intellectual property rights: the possible role of patent pooling. AB - Patent applications that incorporate the genomic sequence of the severe acute respiratory syndrome (SARS) coronavirus, have been filed by a number of organizations. This is likely to result in a fragmentation of intellectual property (IP) rights which in turn may adversely affect the development of products, such as vaccines, to combat SARS. Placing these patent rights into a patent pool to be licensed on a non-exclusive basis may circumvent these difficulties and set a key precedent for the use of this form of mechanism in other areas of health care, leading to benefits to public health. PMID- 16211164 TI - Strategies and approaches in oral disease prevention and health promotion. AB - Oral health is an important element of general health and well-being. Although largely preventable, many people across the world still suffer unnecessarily from the pain and discomfort associated with oral diseases. In addition, the costs of dental treatment are high, both to the individual and to society. Effective evidence-based preventive approaches are needed to address this major public health problem. The aim of this paper is to outline public health strategies to promote oral health and reduce inequalities. An extensive collection of public health policy documents produced by WHO are reviewed to guide the development of oral health strategies. In addition a range of Cochrane and other systematic reviews assessing the evidence base for oral health interventions are summarized. Public health strategies should tackle the underlying social determinants of oral health through the adoption of a common risk approach. Isolated interventions which merely focus on changing oral health behaviours will not achieve sustainable improvements in oral health. Radical public health action on the conditions which determine unhealthy behaviours across the population is needed rather than relying solely on the high-risk approach. Based upon the Ottawa Charter, a range of complementary strategies can be implemented in partnership with relevant local, national and international agencies. At the core of this public health approach is the need to empower local communities to become actively involved in efforts to promote their oral health. PMID- 16211165 TI - The role of the FDI World Dental Federation in global oral health. PMID- 16211167 TI - Leadership with a flexible mentality. PMID- 16211166 TI - I Latin American Guidelines for the Assessment and Management of Decompensated Heart Failure. PMID- 16211168 TI - Management and leadership: analysis of nurse manager's knowledge. AB - Nurses have assumed management positions in many health institutions. To properly accomplish the demands of this role, it is important that they be competent in both management and leadership. For appropriate performance, knowledge of management and supervision styles is a priority. Therefore, the goal of this investigation is to identify the nurse manager's knowledge regarding management and leadership. A structured questionnaire containing twenty-seven questions was applied to twelve Brazilian nurse managers of primary care center called "Family Basic Health Units". Data analysis suggested that the nurse manager lower knowledge in management and leadership is related to visionary leadership, management and leadership conceptual differences, leader's behavior, and situational leadership. And, nurse manager greater knowledge is related to power; team work, and coherence between values and attitudes. PMID- 16211169 TI - [Incidence of pressure ulcers at a university hospital]. AB - This study aimed to identify the incidence of pressure ulcers (PU) at Sao Paulo University Hospital--Brazil, as well as to establish possible associations with patients' sociodemographic and clinical characteristics. Data were collected during three months, involving 211 risk patients, resulting in a 39.8% incidence level. A significant statistical correlation (p<0.05) was found between incidence level and age and PU area; and between age and humidity, time of hospitalization and nutrition, PU area and nutrition. Results indicate the urgent need to implant a PU prevention and treatment program at the institution and provide methodological contributions for other services to develop this kind of research. PMID- 16211170 TI - [Liver transplantation: teaching strategies used with patients waiting for surgery]. AB - This study aims to describe the learning results of the implementation of teaching strategies involving patients who will be submitted to liver transplantation. One of these strategies is to give the patients a manual with orientations and the subsequent application of a questionnaire related to the content of the manual. Authors analyzed 13 patients who were waiting for liver transplantation. With respect to the answers regarding the questionnaire, an average of 83.8% of correct responses was given and only one patient got all the questions right. During the correction and the time to clarify their doubts, authors concluded that the opportunity of reading the manual and applying the questionnaire allowed patients and families to get a better understanding about the surgery's most important aspects. PMID- 16211171 TI - [Postoperative pain in craniotomy]. AB - In the postoperative period, 47% to 75% of the patients report some degree of pain. This study aimed to evaluate pain in the pre and postoperative period of patients submitted to craniotomy. This prospective research was carried out at the neurosurgery unit of a large Brazilian hospital. For a quantitative evaluation of pain, the verbal numeric 0-10 rating scale was used. Forty patients with a mean age of 36 years were evaluated. In the preoperative period, 34 (85%) patients indicated headache as the main cause of pain. In the postoperative period, 37 (93%) patients complained of pain while three (7%) reported absence of pain. Pain peaks were observed on the 2nd postoperative day, when 12 (32%) of the patients reported severe pain and 10 (27%) moderate pain. Absence of severe pain occurred after the 8th postoperative day. It was concluded that protocols of analgesia in craniotomy are needed, such as training nurses to better evaluate and handle pain. PMID- 16211172 TI - Accidents with potentially hazardous biological material among workers in hospital supporting services. AB - Descriptive study was carried out to characterize the occupational accidents involving potentially contaminated material among workers of hospital supporting services. The study reviewed records of workers involved in these accidents and attended at a specialized outpatient clinic of a large tertiary care hospital between January 1997 and October 2001. A total of 2814 workers from different professional categories were attended during this period. Of these, 147 (5.2%) belonged to the hospital supporting services and were the victims of 156 accidents, auxiliary cleaning personnel (80.2%), and over a third of the workers had not received any dose of hepatitis B vaccine (35.4%). Most accidents were due to sharp injuries (96.8%) caused by inadequately discarded hollow needles. Chemoprophylaxis for HIV was not indicated in only 23.1% of cases. We conclude that these workers are also exposed to the possibility of acquiring blood-borne pathogens and that periodical education programs are needed. PMID- 16211173 TI - [Nursing workers' exposure to chemical products at a public universitary hospital in Sao Paulo city]. AB - This study aimed to identify exposure to chemical loads as perceived by nursing workers at the studied institution, disclose their health problems in dealing with chemical products and analyze their suggestions for the decrease or elimination of exposure-related health problems. This research was carried out at the Sao Paulo University Hospital, Brazil--HU-USP. 430 nursing workers were studied. A questionnaire was used for data collection. Results showed that health workers detected 145 chemical substances. The most frequent health problems were related to skin problems (58.14%). 23.72% of the study participants proposed the use of self-protection equipment to decrease or eliminate exposure to these products. PMID- 16211174 TI - [Epidemiological profile of outpatients with HIV/AIDS at a school hospital--Sao Paulo city]. AB - This study aimed to characterize the epidemiological profile of patients with HIV/AIDS at an outpatient service, to analyze the infection stage on the patient's first return, lethality and program evasion. A descriptive and analytical epidemiological study was carried out during nursing appointments with 1837 clients. Most participants were men (1.71 men/1 woman), between 30 and 39 years old (46.8%); residents in Sao Paulo (86.5%); low education level (76.3%). Most women were living with partners (45.4%), whereas most men were single (53.3%). The main category of exposure was heterosexual behavior (83.5% among men and 71.2% among women). Both male and female participations discovered their HIV/AIDS positive condition in advanced stages (57%). Opportunistic infection levels were significantly higher (p < 0.05) among men. For men and women, the predominant opportunistic infection was tuberculosis (26.2%). CD4 < 350 results on the first return were significantly higher (p = 0.04) among women. Death risk was greater among women (OR = 1.72), whereas the risk of treatment evasion was greater among men (OR = 1.72). PMID- 16211175 TI - [Microbiological evaluation of 1% silver sulfadiazine dressings]. AB - This study aimed at evaluating the microbial condition and antimicrobial activity of 1% silver sulfadiazine dressings used in burns, prepared in advance of the dressing change and stored for 12 to 60 hours before usage. The microbial condition was evaluated by means of three cultures prepared from the Letheen Broth (Difco) culture medium and incubated at 32-35 degrees C for 20 days. Antimicrobial activity was evaluated by means of the diffusion technique of one dressing culture in hospital strains, initially incubated at room temperature and then at 37 degrees C, for 18-24 hours. All three cultures of 12, 36 and 60 hours proved negative. Concerning antimicrobial activity, the zones of inhibition were: 2.25 mm for S. aureus; 2.65 mm for P. aeruginosa and 2.95 mm for E. coli. These dressings can be safely prepared by nurses and stored, reducing nurses' time spent for dressing change and, consequently, patients' exposure. PMID- 16211176 TI - [Nursing students' perceptions on HIV serodiscordant partnerships]. AB - This qualitative research aimed to identify undergraduate nursing students' perceptions on mixed-HIV-status couples. Social Representation Theory was used to get to know how the students feel, think and act towards HIV/aids serodiscordance. Six fourth-year nursing students were interviewed. Participants were between 20 and 26 years old. The "Projective Thematic Drawing" and a structure interview were used for data collection. Data were analyzed by means of "Thematic Content Analysis". The obtained data revealed the students' perceptions on serodiscordant couples. This study triggered future reflections/discussions on health education for mixed-HIV-status couples and nursing care. PMID- 16211177 TI - [Epidemiology of accidents and violence against children in a city of Southern Brazil]. AB - This study aimed to analyze the epidemiological characteristics of accidents and violence against children under 15 years old who lived in Londrina, a city in the South of Brazil, in 2001. Morbidity data were collected from general hospitals records and mortality data were obtained from the Municipal Mortality Information Center. A total of 8,854 children were studied, which corresponded to an incidence rate of 74.8/1,000 children. Admission and fatality rates were 4.2% and 0.2%, respectively. The incidence rate was higher among two-year olds (109/1,000) and male victims prevailed (60.7%). Other causes of injuries were the main subtype of external cause (61.0%), followed by events of undetermined intention (30.6%) and by transport-related injuries (7.5%). The head was the most affected body part (34.9%) and superficial injuries were the most frequent kind of trauma (32.4%). The results contribute to the planning of injury control and prevention actions. PMID- 16211178 TI - [Gerontological training of nursing technicians: a cultural approach]. AB - Brazilian nursing has been trying to discuss social-political questions that interfere in health and higher as well as technical education. One of the challenges posed nowadays is the population's aging, which occurs at an accelerated rate and demands redefinition of roles and actions to attend to the elderly population. This case study aimed to understand the gerontological training of nursing technicians from a cultural approach, using a qualitative methodology. Results revealed deficiencies in the formation of competent nursing technicians who are trained in elderly care. Human beings' use of symbols and meanings of past experiences was seen as a reference point for the promotion of transforming initiatives that correspond to social reality. PMID- 16211179 TI - [Perception of the family health care team about the use of the Basic Health Care Information System-SIAB]. AB - This study aimed to identify and analyze the use of the Basic Health Care Information System-SIAB by the family health care team, identifying the aim, possible difficulties and facilities of using these forms. This is an exploratory, descriptive study with a qualitative approach. We have used semi structured taped interviews with the family health care team in Franca-Sao Paulo- Brazil. The analysis was structured into four groups: the usage of SIAB as a work instrument for the health care team; contradictions in the usage of SIAB; limitations as to data input; and suggestions on how to solve these problems. We will show that the team's use and involvement with SIAB is sporadic, due to difficulties in analyzing, monitoring and evaluating data for complying with local programs related to community health agents, whose high turnover directly affects the inadequate completion and interpretation of forms, as well as difficulties to understand the aim of these forms. PMID- 16211180 TI - [Interdisciplinary home hospitalization program of Marilia-SP: material resource costs]. AB - This study estimated the mean total direct cost of some production factors (material resources), including consumption materials, diets, medication and solutions used by two patient groups during hospitalization in the Interdisciplinary Home Hospitalization Program (PROIID) of Marilia-SP, Brazil. The sample consisted of 27 patients with a diagnosis of cerebral vascular accident (CVA) and 18 with Neoplasm. Data were collected from the moment of hospitalization until the patients left the program. The mean total direct cost amounted to R$ 52.72 for each patient with CVA and of R$ 65.13 for those with Neoplasm. For both patient groups, the sub-category of material--diets accounted for the highest percentage of costs. PMID- 16211181 TI - [Living the mother-child-family triad in the Kangaroo Method]. AB - This study focuses on the experience of families in the Kangaroo Mother Method (KM) and aims to understand the functioning of the family dynamics and to apprehend the family transformations as a result of their experience in the method. The study used Symbolic Interactionism as a theoretical and Grounded Theory as a methodological reference framework. Data revealed the following phenomena: having an unexpected evolution and outcome of pregnancy, which is a phase in which the parents go through a pregnancy and suffer due to its premature interruption; coping with the prematurity of the child, in which the parents get into the world of prematurity and face the need to act for the child's survival; and living with the decision and the experience together with the child, which reveals the possibility of staying in hospital in the KM or not. These phenomena led to the central category: weighing the risks and benefits between staying with the child in the kangaroo method or with the family. PMID- 16211182 TI - [Human care: an ethical reflexion on HIV/AIDS patients]. AB - This descriptive-reflexive study reflects on the importance of ethics in human care and its relation with HIV/AIDS patients, identifying the main concepts of ethics in human care and ethics in care for HIV/AIDS patients. Considering the clear need for care nowadays, current ideas present care as the human essence which is present in ethics. The emergence of AIDS, an epidemic that cannot be cured yet, mobilizes society to concerns about privacy and confidentiality, including the need to reinforce ethical aspects, with a view to a better life quality for its patients. Ethical thinking is in line with scientific and technological evolution all over the world. In view of our reflections, we understood that ethics is part of our society and that human care needs to be linked up with ethics. PMID- 16211183 TI - [Health-sickness-care process and the street population]. AB - This paper gives an empirical and contextualized presentation of the health sickness-care process and its relation with poverty, emphasizing the problem of people who live in and of the street. We try to stimulate a reflection on the technical, scientific and political function of nursing care for the street population. This paper is structured in three topics: first, we present the problem of people living in the streets of Sao Paulo city, characterizing their peculiarities and emphasizing ways of survival. Next, we situate the theoretical study area, discussing the health-sickness-care process and its relation with poverty, as well as nursing care. Finally, we present some final considerations. PMID- 16211184 TI - [Geertz' Interpretive Theory and care management: visualizing nurses' social practice]. AB - This paper presents a theoretical reflection on hospital nursing care management and Geertz' Interpretive Theory of Culture. We discuss some significant elements of culture in management, based on the theoretical reference frameworks of nursing, administration and anthropology. In these, the importance of cultural diversity is highlighted as an innovative resource to expand the vision of human integrity, valuing divergences, respect and sharing, which are important for nurses in the construction of their social practice. PMID- 16211185 TI - [About the concept of madness and its reflexes on mental health care]. AB - This is a theoretical reflection on the historical perception and conceptualization of madness and, consequently, on ways of acting towards it. We start from ancient Greek conceptions and move up to the present days, focusing mainly on those periods when perceptions and actions towards madness changed. We conclude that getting to know the historical forms of the madness concept is important to be able to denaturalize our conceptions and reflect about our current practice in care for patients going through mental suffering. PMID- 16211186 TI - [Pain in undernourished children: the mother's perception]. AB - This qualitative study aimed to identify how mothers perceive pain in their undernourished children. Semistructured interviews were realized at the Institute for the Prevention of Malnutrition and Exceptionality--IPREDE (Fortaleza-Ceara Brazil). Participants were mothers who accompanied their undernourished children. Data analysis revealed the following categories: Coming to the Institution; Pain description and How to take care of the pain. Society needs to take actions, in respect of citizens' rights and culture, with a view to reverting this picture of pain in undernourished children. PMID- 16211187 TI - Treating overweight children and adolescents in the clinic. AB - The increasing prevalence of pediatric overweight has caused the medical community to begin searching for ways to deal with this new pediatric medical problem. The Centers for Disease Control developed the Body Mass Index (BMI) growth charts, which came into use in 2000. Primary care providers are seeking education on this relatively new topic. This article provides fundamental information based on the medical evidence for pediatricians to learn how to care for their overweight pediatric patients in the office setting. PMID- 16211188 TI - Pediatric allergic rhinitis: antihistamine selection. PMID- 16211189 TI - Ear and nose foreign bodies: "It is all about the tools''. PMID- 16211190 TI - Difficulty in defecation in infants with gastroesophageal reflux treated with smaller volume feeds thickened with rice cereal. AB - We prospectively evaluated the incidence of difficulty in defecation in infants with gastroesophageal reflux who were treated with smaller volume feeds thickened with rice cereal and also assessed the effect of changing the cereal to oatmeal. We evaluated 53 thriving infants with uncomplicated gastroesophageal reflux who were treated with smaller volume feeds thickened with rice cereal. Parents maintained records of bowel movements for 7 days. Rice was substituted by oatmeal cereal in those infants developing difficulty in defecation and another 7 days' record was kept. Of the 53 infants enrolled, 34 (64%) reported no difficulty in defecation, 8 (15%) reported mild difficulty, and 11 (21%) reported severe difficulty in defecation during rice-based feedings. In these symptomatic 19 infants, after rice was substituted by oatmeal cereal, 10 infants (52.6%) reported no symptoms, 6 (31.6%) had mild symptoms, and 3 (15.8%) continued to have severe symptoms. We conclude that difficulty in defecation is common during treatment of infants with gastroesophageal reflux with smaller volume feeds thickened with rice cereal. Substitution of rice with oatmeal cereal results in partial or complete resolution of symptoms in most of these infants. PMID- 16211191 TI - Suppurative complications of frontal sinusitis in children. AB - A retrospective review of children diagnosed and treated for suppurative complications of paranasal sinusitis was undertaken to describe clinical presentation, microbiology, and treatment. This review includes children with subgaleal abscess and osteomyelitis of the frontal bone, subdural empyema, frontal lobe abscess, meningitis, and encephalitis. Staphylococcus aureus and group C beta-hemolytic Streptococcus were isolated agents. All children were treated with intravenous antibiotics with drainage of both the sinus and extracranial and intracranial suppurations. Results of treatment in the series support the opinion that combined aggressive surgical and antibiotic treatment is a preferred method in complicated sinusitis in children. PMID- 16211192 TI - Vitamin D insufficiency in preadolescent African-American children. AB - To determine the proportion of vitamin D insufficiency in 6- to 10-year-old preadolescent African-American children residing in Pittsburgh, Pennsylvania and to estimate their therapeutic response to vitamin D 400 IU/day for 1-month, an open-label pre- and post-comparison of vitamin D status following vitamin D 400 IU daily for 1 month during winter and early spring was conducted. Outcomes included serum calcium, phosphorus, albumin, 25 hydroxyvitamin D [25 (OH) D], 1, 25 dihydroxyvitamin D [1, 25 (OH) (2) D], parathyroid hormone (PTH), and markers of bone turnover (serum bone-specific alkaline phosphatase, osteocalcin, and urine n-telopeptide crosslinked collagen type 1 [NTX]). Dietary intake of vitamin D was assessed using a food frequency questionnaire. Forty-one of the 42 enrolled subjects (mean age: 8.9 +/- 1.2 yrs [SD]) were analyzed, and 20/41 (49%) were vitamin D insufficient. Vitamin D insufficient group had a suggestive trend of being older (9.2 +/- 1.0 years vs. 8.5 +/- 1.3 years, p = 0.06) and more pubertally advanced (Tanner II: 7/20 vs. Tanner II: 1/21, p = 0.02). Mean dietary intake of vitamin D was 277 ( 146 IU/day (n = 41). Adequate intake for vitamin D (200 IU/day) was not met in 16/41 (39%); however, the dietary intake of vitamin D was not significantly different between the vitamin D insufficient and vitamin D sufficient groups. PMID- 16211193 TI - Does crying turn tympanic membranes red? AB - The diagnosis of acute otitis media is based on several clinical factors. One of these factors is the color of the tympanic membrane (TM). Crying can cause flushing and hyperemia of the face. The purpose of this study is to determine whether crying affects the color of tympanic membranes. Infants and toddlers (age 30 months or less) evaluated in an outpatient clinic or primary care pediatrician's office for routine well-baby checks who received at least 2 parenteral immunizations were enrolled on a convenience basis. Ill children were excluded. The initial physician assessed crying and TM color/visibility. Following immunizations, a second physician assessed crying and the TM color/visibility. Color differences were stratified by the degree of crying. One hundred twenty-one study subjects received 2-5 parenteral immunizations. TM colors were most often in the pink range or less. Only 2 TMs were assessed as light red and none were assessed as red. Twenty-eight percent of the TMs with greater crying on the second exam were noted to be redder on the second exam compared to the first exam versus 11% for the comparison group (p=0.0007); 19% of the TMs with greater crying on the second exam were noted to be redder by 2 or more increment levels compared to the first exam versus 5% for the comparison group (p=0.0004); 31% of the TMs with greater crying on the second exam at the 3+ and 4+ level were noted to be redder on the second exam compared to the first exam versus 14% for the comparison group (p=0.003). Our data indicate that, in some instances, crying can result in an increase in pinkness of the TM. Crying in well children does not result in a red tympanic membrane. PMID- 16211194 TI - Human bites and blood exposures in New York City schools. AB - The epidemiology of human bites and blood exposures in urban schools has not been described. The authors reviewed 734 human bite and blood exposure incidents in New York City schools from September 1999 to June 2001. School site, age, gender, body site, temporal distributions, and underlying health status of individuals were reviewed. Nine incidents involved children known or suspected to be HIV infected. Medical interventions included EMS involvement, emergency room treatment, hospitalization, vaccine administration, laceration suturing, systemic antibiotic therapy, and serologic evaluation for transmission of blood-borne pathogens. These incidents occur with a high prevalence in NYC schools and have potential for serious outcomes. PMID- 16211195 TI - Hepatitis A immunization strategies: universal versus targeted approaches. AB - The alternative to nationwide childhood hepatitis A vaccination is to continue targeting high-risk adults. To consider how many of today's children will become vaccination candidates, the proportion of adults reporting a lifetime history of hepatitis A risk factors was examined. One thousand thirty-four US adults responded to a confidential postal survey, 49% of whom had met current Advisory Committee on Immunization Practices (ACIP) criteria for immunization and 72% had met an expanded set of criteria. Because adult vaccination is more costly per capita, targeted vaccination may provide modest financial savings with none of the benefits associated with reduced child-to-adult transmission. PMID- 16211196 TI - Screening for hypertension in high school. AB - Arterial hypertension is a major risk factor for atherosclerotic ischemic heart disease and cerebrovascular stroke. Blood pressure measurements were taken in the supine position in 6,282 healthy adolescents (3073 boys, 3209 girls) aged 13-17 years of both sexes. Forty-eight subjects were found to have hypertension: 35 idopathic and 13 secondary to reflux nephropathy, chronic glomerulonephritis, coarctation of the aorta, and hemolytic uremic syndrome. Systolic blood pressure was significantly higher among males than females at ages 15-17 years (p<0.001); diastolic blood pressure was higher among males at age 17 years only (p<0.05). Males with BMI>17 exhibited higher systolic blood pressure than females. Prevalence of hypertension, mostly primary, in a large cohort of students was 0.76%. Early screening is important for early interventions and reducing cardiovascular morbidity and mortality in adulthood. PMID- 16211197 TI - Presenting signs and symptoms of pheochromocytoma in pediatric-aged patients. AB - To describe the varied presenting signs and symptoms in pediatric patients with pheochromocytoma, a retrospective chart review of the presenting signs and symptoms and subsequent clinical course of patients who presented to the Pediatric ICU following surgical excision of a pheochromocytoma was undertaken. The cohort of 7 patients (5 boys, 2 girls) ranged in age from 4 to 16 years. Two patients were hypertensive at initial presentation, and the other 5 developed hypertension after their initial presentation. The initial presenting signs and symptoms were related to the central nervous system (CNS) in 6 of the patients (5 with an acute alteration in mental status and 1 with visual disturbances). Two patients presented with congestive heart failure. Other signs and symptoms at the time of initial presentation included sweating, headache, weight loss, heat intolerance, increased thirst and urination, a decline in school activity, and red/puffy hands and feet. The time from the initial presentation until the diagnosis was confirmed was 5 months or more in 4 of the 7 patients. The diagnosis was confirmed by demonstration of elevated urinary catecholamines in all 7 patients, although 2 patients had initial negative urinary levels. PMID- 16211198 TI - Kawasaki disease hospitalizations in a predominantly African-American population. AB - This is a descriptive study of the occurrence of Kawasaki disease (KD) in an urban population that was a majority of African Americans. Records of 189 children (mean age, 2.9 +/- 2.2 years [range: 2 months to 11.1 years]) hospitalized for KD over 8 years (January 1, 1992 to December 31, 1999) were reviewed and data analyzed. One hundred thirty-six (72%) were African American (AA), 43 (23%) were white, and 9 (5%) children were "others.'' The annual frequency was 15 for AA and 7.7 for white per 100,000 5-year-old children. Coronary artery abnormalities (CAA) were reported in 21 (11%) children (18 [13.2%] of 136 AA, and 3 [4.7%] of 43 whites [p=0.095]). AA children with CAA were older than their white counterparts (26 to 24 vs. 5 to 2.8 months, p=0.03). There was a higher occurrence in winter and spring (110 cases [58%] vs. 79 cases [42%]) compared to summer and fall. KD occurrence was positively associated with average monthly snowfall (r=0.35, p=0.004) and inversely associated with average monthly temperature (r = - 0.2, p=0.048). African-American children were more likely to be hospitalized for KD compared to white children. The association of KD with temperature and precipitation suggest that it is influenced by environmental factors. PMID- 16211199 TI - Pervasive developmental disorders: consideration of developmental level for diagnosis. PMID- 16211200 TI - Ballet dancer with hip and groin pain: Crohn disease and psoas abscess. PMID- 16211201 TI - An inappropriate diet. PMID- 16211202 TI - Prolongation of repolarization as antifibrillatory action revisited: drug combination therapy in atrial fibrillation. PMID- 16211203 TI - Review of antiplatelet therapy in secondary prevention of cerebrovascular events: a need for direct comparisons between antiplatelet agents. AB - Patients experiencing stroke or transient ischemic attack (TIA) are at high risk for recurrent (secondary) strokes, which comprise 29% of all strokes in the United States. Current recommendations for prevention of secondary stroke from the American College of Chest Physicians (ACCP) call for the broad use of platelet antiaggregation (antiplatelet) agents for patients with a history of noncardioembolic stroke or TIA. Five agents--aspirin, ticlopidine, clopidogrel, extended-release dipyridamole (ER-DP), and triflusal--have demonstrated efficacy in large-scale clinical studies in the prevention of recurrent vascular events and/or stroke in patients with a history of stroke. The results of the following studies are reviewed and compared: the Swedish Aspirin Low-Dose Trial (SALT), the United Kingdom Transient Ischaemic Attack (UK-TIA) Aspirin Trial, Dutch Transient Ischemic Attack (Dutch TIA) study (aspirin), the Canadian American Ticlopidine Study (CATS), the Ticlopidine Aspirin Stroke Study (TASS), the African American Antiplatelet Stroke Prevention Study (AAASPS) (ticlopidine), the Clopidogrel versus Aspirin in Patients at Risk of Recurrent Ischemic Events (CAPRIE) trial, the Management of Atherothrombosis With Clopidogrel in High-Risk Patients study (MATCH) (clopidogrel), the second European Stroke Prevention Study (ESPS2) (aspirin plus ER-DP), and the Triflusal versus Aspirin in Cerebral Infarction Prevention (TACIP) study. In comparative monotherapy studies of patients with previous stroke, ticlopidine demonstrates statistically significant improved efficacy over aspirin, and clopidogrel demonstrates nonsignificant slight improvement over aspirin for the prevention of ischemic cardiac and cerebrovascular events; however, the adverse event profile of ticlopidine (including rash, diarrhea, and neutropenia) will probably limit its long-term use. Among combination approaches, only aspirin plus ER-DP has demonstrated statistically significant, clinically meaningful additive benefit over monotherapy with each agent. Clopidogrel plus aspirin did not significantly improve preventive efficacy and increased the risk of serious side effects, including life-threatening bleeding episodes. The 15,500-patient PRoFESS (the Prevention Regimen for Effectively Avoiding Second Strokes) study, with results expected in 2008, will directly compare aspirin plus ER-DP with clopidogrel monotherapy for the prevention of recurrent stroke and should provide statistically robust estimates of comparative efficacy for the development of improved recommendations. PMID- 16211204 TI - Hypothesis: antiplatelet effects of selective serotonin reuptake inhibitors cause clinical benefits on cardiovascular disease and increase risks of bleeding. PMID- 16211205 TI - Tolvaptan administration does not affect steady state amiodarone concentrations in patients with cardiac arrhythmias. AB - BACKGROUND: Tolvaptan, a nonpeptide selective vasopressin receptor (V2) antagonist, is in development for the treatment of congestive heart failure and hyponatremia. Tolvaptan is primarily metabolized via CYP3A4. This study was conducted to determine the extent of the pharmacokinetic interaction between tolvaptan and steady state amiodarone, an antiarrhythmic drug commonly prescribed for patients with congestive heart failure and a known inhibitor of other drugs metabolized by CYP3A4. METHODS: This was a multicenter, open-label, 1-arm, 3 period, sequential treatment study conducted in 11 men (10) and women aged 49 to 80 years. They were primarily Caucasian (20) subjects, with a history of cardiac arrhythmias who were otherwise healthy. Subjects were to have been on oral amiodarone maintenance therapy of 200 mg/day for at least 10 months. All subjects took 200 mg amiodarone once daily on each study day; on days 3 and 4, they were also coadministered 30 and 90 mg of tolvaptan, respectively. The plasma concentrations of amiodarone and its metabolite desethylamiodarone were determined for 24 hours postdose on days 2, 3, and 4, tolvaptan concentrations were determined for 24 hours postdose on days 3 and 4. RESULTS: As determined by the ratio of the geometric means and 90% confidence intervals (0.5 to 2.0) for the maximal plasma concentration and the area under the curve during the dosing interval for both amiodarone and desethylamiodarone, tolvaptan coadministration had no effect on either amiodarone and desethylamiodarone disposition, as all the geometric mean ratios (amiodarone + tolvaptan [30 or 90 mg] vs amiodarone alone) were approximately 1. CONCLUSION: Tolvaptan coadministration does not alter steady-state amiodarone or desethylamiodarone concentrations. Tolvaptan concentrations did not appear to be different from historical controls. The most frequently reported adverse event was polyuria (15 of 21 subjects for amiodarone + 30 mg tolvaptan); an expected outcome due to the known potent aquaretic action of tolvaptan. The combination of amiodarone and tolvaptan was well tolerated. PMID- 16211206 TI - Simple clinical criteria to determine the prognosis of heart failure. AB - OBJECTIVE: To determine whether easily obtained clinical parameters serve as predictors of survival in patients with congestive heart failure. Several scoring systems to predict heart failure survival have been developed; however, many of these deal principally with transplant recipients or do not account for a patient's response to therapy. METHODS: A total of 680 patients with an ejection fraction of less than 40% were included in the analysis. Baseline assessments were performed and treatment regimens were identified; patients were then followed for up to 5 years. Univariate and multivariate Cox regression models were used to determine clinically important predictors of survival. Kaplan-Meier survival functions for patients with and without the prognostic variable were constructed and mortality was calculated at 1 year and 5 years. RESULTS: Ejection fraction improvement at 6 months, diabetes mellitus, age, serum creatinine, and blood urea nitrogen (BUN) were significant predictors for survival in the univariate model. Ejection fraction improvement, age, and BUN were significant predictors in the multivariate model. These findings were used to construct a model for predicting patient mortality. Improved ejection fraction (>15 ejection fraction units) gave a 1-year mortality of 2% and a 5-year mortality of 11%. Mortality rates according to patient age and BUN levels were also calculated. CONCLUSION: Ejection fraction improvement was the most important predictor for survival in patients with systolic dysfunction; monitoring ejection fraction changes through repeat echocardiograms has important prognostic value. In patients without ejection fraction improvement, age and renal function are important survival determinants. PMID- 16211207 TI - Carvedilol improves myocardial contractility compared with metoprolol in patients with chronic hibernating myocardium after revascularization. AB - BACKGROUND: We tested the hypothesis of whether carvedilol delays morphologic degeneration and improves functional outcome compared with metoprolol tartrate in patients with hibernating myocardium undergoing surgical revascularization. We have previously shown that patients with chronic hibernating myocardium undergo progressive cellular degeneration and fibrosis. METHODS: Twenty patients with multivessel coronary artery disease revascularization and hibernating myocardium as assessed by technetium-99m perfusion scintigraphy and fluorine-18 fluorodeoxyglucose positron emission tomography were randomized to receive either carvedilol or metoprolol tartrate for at least 2 months before surgery, and this was continued for 7 months postoperatively. Left ventricular ejection fraction and regional wall motion abnormalities were assessed by left ventriculography at baseline and 7 months postoperatively. Intraoperative transmural needle biopsy samples were obtained for microscopic analysis. RESULTS: Postoperatively, the ejection fraction increased from 31% +/- 5% to 44% +/- 4% (P < .005) in the carvedilol group (n = 10), and from 30% +/- 6% to 40% +/- 6% in the metoprolol tartrate group (P < .05 vs preoperatively and vs carvedilol). Wall motion abnormalities in the carvedilol group improved from -2.1 +/- 0.4 to -0.6 +/- 0.5 (P < .05) and from -2.3 +/- 0.5 to -1.6 +/- 0.6 in the metoprolol tartrate group (P < .05 vs preoperatively and vs carvedilol). Microscopic analysis after 72 +/- 18 days of either treatment showed mild cardiomyocyte degeneration and moderate to-severe fibrosis (28% +/- 7%) in the carvedilol group compared with moderate cardiomyocyte degeneration and moderate-to-severe fibrosis (33% +/- 6%) in the metoprolol tartrate group. Apoptosis, as assessed by the terminal deoxynucleotidyl transferase nick end labeling method, was observed in only 1 patient in each group. CONCLUSIONS: Carvedilol treatment of hibernating myocardium results in improved functional recovery after revascularization compared with metoprolol tartrate, and this might partially be related to reduced cardiomyocyte degeneration. PMID- 16211208 TI - Marked QT prolongation and torsades de pointes secondary to acute ischemia in an elderly man taking dofetilide for atrial fibrillation: a cautionary tale. AB - Dofetilide has been shown to be effective and safe in maintaining sinus rhythm in patients with persistent atrial fibrillation and congestive heart failure. Because of serious side effects of an increase in the QT interval causing torsades de pointes, dofetilide must be initiated with close monitoring of the QT interval in an inpatient setting. However, little has been reported about conditions surrounding the change in QT interval after the steady state is achieved that may have implications in the safety and efficacy of the drug. We report marked QT prolongation and torsades de pointes in a setting of flash pulmonary edema resulting from acute myocardial ischemia in a patient who was being treated with dofetilide for atrial fibrillation. Our case reminds the clinicians that the adverse and proarrhythmic effects of dofetilide can occur due to changes in the arrhythmic substrate during acute severe ischemia. PMID- 16211209 TI - Beneficial cardiovascular actions of eplerenone in the spontaneously hypertensive rat. AB - BACKGROUND: Aldosterone has been implicated as a potential mediator of cardiac and vascular damage in a variety of disorders. This study examined the role of aldosterone and its interplay with the renin-angiotensin system in the pathogenesis of hypertension. To this end, the effects of the aldosterone antagonist eplerenone and the angiotensin converting enzyme inhibitor lisinopril on cardiovascular mass, myocardial collagen, and coronary circulation were examined in spontaneously hypertensive rats. METHODS: Male, 22-week-old rats were randomly divided into 4 groups (12 in each). The control group received no treatment, the second group was given eplerenone (100 mg/kg/day), the third received lisinopril (3 mg/kg/day), and the fourth was given eplerenone and lisinopril. After 12 weeks of respective treatments, systemic and regional hemodynamics and cardiovascular mass indexes were measured in conscious instrumented rats. RESULTS: Eplerenone decreased arterial pressure but did not affect left ventricular mass or hydroxyproline concentration (an estimate of collagen). It did, however, reduce minimal coronary vascular resistance and increased coronary flow reserve. Lisinopril decreased arterial pressure and ventricular mass but did not affect regional hemodynamics. The combination therapy produced synergistic effects. CONCLUSION: Aldosterone antagonism improved coronary and systemic hemodynamics in adult spontaneously hypertensive rats but did not affect cardiovascular mass indexes. The finding that lisinopril and eplerenone decreased arterial pressure to the same extent but had different cardiovascular effects suggested that these effects might be pressure independent. PMID- 16211210 TI - Effectiveness of oral magnesium in a patient with ventricular tachycardia due to hypomagnesemia. AB - A 12-year-old girl with occasional symptoms of chest discomfort was diagnosed with ventricular tachycardia on cardiac evaluation. No evidence of organic heart disease was apparent, but a laboratory evaluation revealed hypomagnesemia. Ventricular tachycardia disappeared after treatment with 200 mg/day of oral magnesium hydroxide, and no further chest discomfort was reported. In addition to routine cardiac evaluation for arrhythmia, serum magnesium levels should be checked in patients with suspected idiopathic benign ventricular tachycardia. Treatment with oral magnesium is a physiologic therapy and should be considered for patients with ventricular tachycardia due to hypomagnesemia. PMID- 16211211 TI - 5E, 8Z, 11Z, 14Z-eicosatetraenoic acid, a novel trans isomer of arachidonic acid, causes G1 phase arrest and induces apoptosis of HL-60 cells. AB - Trans arachidonic acid isomers (trans-AA) constitute a new group of trans fatty acids (trans-FA) generated in vivo via endogenous cis-trans isomerization stimulated by the NO2 radical. Because both NO2 and trans-FA have been implicated as causative factors in cancer, we studied the effect of the trans-AA isomers on proliferation and viability of human promyelocytic (HL-60) cells. The four trans arachidonic (trans-AA) acid isomers synthesized by us have been presently tested with respect to their competence to affect the proliferation and viability of human promyeolocytic HL-60 cells in culture. The data demonstrate that one of the isomers, 5,6-trans-AA, showed distinct activity by targeting cell progression through the cell cycle and inducing apoptosis. The effects were time- and concentration-dependent: the cytostatic effect of 5E-AA was observed at 10 microM following 72 h of treatment. This effect was manifested as a perturbation of cell progression through G1 phase, indicating the 'on' activation of the G1 checkpoint as evidenced by the flow- and laser scanning-cytometry techniques. Apoptotic cells were identified by comparison of their morphology, DNA fragmentation, caspase activation and collapse of mitochondrial potential with control cells. These observations suggested that 5E-AA induced a mitochondrial pathway of apoptosis. There was no evidence of cell-cycle phase specificity in induction of apoptosis by 5E-AA, as the cells showing highly fragmented DNA or caspase-3 activation were distributed in all phases of the cycle. The data suggest that 5E AA may have at least two targets: one that is cell-cycle specific and associated with the observed arrest in the G1 phase and another, unrelated to the cell cycle, which is responsible for triggering apoptosis indiscriminately, regardless of cycle phase I. PMID- 16211212 TI - FISH-mapping and genomic organization of the NAD-dependent histone deacetylase gene, Sirtuin 2 (Sirt2). AB - Sirtuin 2 (SIRT2) is a nicotinamide adenine dinucleotide (NAD(+))-dependent deacetylase, which belongs to the Silent information regulator 2 (Sir2) family of sirtuin histone deacetylases (HDACs). The yeast Sir2 protein and its mammalian derivatives play a central role in epigenetic gene silencing, DNA repair and recombination, cell-cycle, microtubule organization, and in the regulation of aging. We have isolated and characterized the human Sirt2 genomic sequence, which spans a region of 20,960 bp and which has one single genomic locus. Determination of the exon-intron splice junctions found the full-length SIRT2 protein to consist of 16 exons ranging in size from 16 bp (exon 1) to 749 bp (exon 9). The 1,963-bp human Sirt2 mRNA has an open reading frame of 1,167 bp that encodes two isoforms of the SIRT2 protein: isoform 1 encodes a 389-aa protein with a predictive molecular weight of 43.2 kDa and an isoelectric point of 5.22, while isoform 2, which is lacking the first three exons, encodes a 352-aa protein with a predictive molecular weight of 39.5 kDa and an isoelectric point of 6.05. Characterization of the 5' flanking genomic region, which precedes the Sirt2 open reading frame, revealed a TATA- and CCAAT-box less promoter that contains a 0.67 kb CpG island and a number of NFkappaB and GATA transcription factor binding sites. Fluorescence in situ hybridization analysis localized the human Sirt2 gene to chromosome 19q13.1. Human SIRT2 is known to be most predominantly expressed in the brain. Since it is however severely reduced in a large number of human brain tumor cell lines, the absence of SIRT2, a potential tumor suppressor, could play a key role in the regulation of the cell-cycle within a multistep pathway that leads to full cellular transformation and, finally, the development of cellular malignancy. PMID- 16211213 TI - Ephrin-B2 overexpression enhances integrin-mediated ECM-attachment and migration of B16 melanoma cells. AB - Eph-receptor tyrosine kinases (Eph-RTKs) and their membrane-bound receptor-like ligands, the ephrins, represent a cell-cell signaling system that directs cellular migration during development. Differential expression in cancer suggests similar roles in tumor progression. We have previously shown that ephrin-B2 mRNA is overexpressed in advanced malignant melanomas (MM). In this study, immunohistochemistry revealed a most prominent expression of ephrin-B2 in the invasive front of advanced MM. Therefore, we addressed the question of whether ephrin-B2 signaling modulates MM cell migration and matrix interaction. Using a wild-type ephrin-B2-negative B16 mouse MM subclone we show that overexpression of ephrin-B2 leads to the formation of multiple lamellipodia, enhanced polymerisation of actin fibers, and induction of focal adhesion complexes with constitutive activation of focal adhesion kinase. Consequently, ephrin-B2 overexpressing B16 cells display a significant increase of beta1-integrin mediated attachment to matrix components, preferentially laminin and fibronectin. As a further effect of ephrin-B2 overexpression, we observed an accelerated migration in both Boyden chamber invasion experiments as well as in in vitro scratch-wound assays. We conclude that ephrin-B2 can act as a major modulator of cell migration and matrix interactions of MM cells, which possibly contributes to the expansion and metastatic spread of MM in vivo. PMID- 16211214 TI - Selective accumulation and strong photodynamic effects of a new photosensitizer, ATX-S10.Na (II), in experimental malignant glioma. AB - We investigated the feasibility of a novel photosensitizer, ATX-S10.Na (II), in photodynamic therapy (PDT) for glioma. First, PDT was performed in various brain tumor cell lines in vitro. Cytotoxicity depended upon both drug concentration and laser energy and the 50% inhibitory concentration ranged from 3.5 to 20 microg/ml. Next, PDT was performed in the subcutaneous and intracranial 9L tumor models in Fischer rats using ATX-S10.Na (II) and light from a 670-nm diode laser delivered by intratumoral insertion of an optical fiber. The effect of PDT on brain tumors was evaluated using magnetic resonance imaging. Sequential changes of the ATX-S10.Na (II) concentrations were also measured quantitatively by fluorospectrometry up to 12 h after intravenous administration in rats with intracranial and subcutaneous tumors. The concentration of ATX-S10.Na (II) in the brain tumor reached a maximum at 2 h after administration and the tumor/normal brain concentration ratio was as high as 131 at 8 h. Intratumoral PDT for intracranial tumors irradiated at this timing showed an obvious anti-tumor effect without severe side effects. The present study demonstrated the highly selective accumulation of ATX-S10.Na (II) in tumor tissue and its potent photodynamic effect in an experimental malignant glioma model. PMID- 16211215 TI - Survivin gene expression is negatively regulated by the p53 tumor suppressor gene in non-small cell lung cancer. AB - Survivin is considered to be associated with tumorigenesis by regulating apoptosis and cell proliferation. Recent experimental studies reported survivin gene expression to be negatively regulated by wild-type p53. We investigated resected tumor specimens from 140 non-small cell lung cancer (NSCLC) patients. Quantitative reverse-transcription PCR analysis was performed to evaluate survivin gene expression. PCR-single strand conformation polymorphism following sequencing was performed to investigate mutations of p53. The apoptotic index and the Ki-67 proliferation index were also evaluated according to the survivin expression. The survivin expression was low in normal lung tissue. In contrast, the survivin expression varied greatly among tumor tissues. The survivin expression in squamous cell carcinomas was significantly higher than that in adenocarcinomas (P=0.0109). The survivin expression in moderately or poorly differentiated tumors was significantly higher than that in well-differentiated tumors (P=0.0334). Furthermore, the survivin expression in tumors with mutant p53 was significantly higher than that in tumors with wild-type p53 (P=0.0026). In addition, the apoptotic index was significantly lower in high-survivin tumors than in low-survivin tumors (P<0.0001). The Ki-67 proliferation index was significantly higher in high-survivin tumors than in low-survivin tumors (P<0.0047). This study indicated survivin gene expression to be negatively regulated by p53 in NSCLC, and that survivin expression could inhibit apoptosis and accelerate tumor cell proliferation to produce more aggressive carcinomas. PMID- 16211216 TI - IGFBP-3 expression in hepatocellular carcinoma involves abnormalities in TGF-beta and/or Rb signaling pathways. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) is a mediator of growth suppression signals and a putative tumor suppressor gene. The growth suppression mechanisms of IGFBP-3 have not been well clarified. We examined the expression of IGFBP-3 transcripts in human hepatocellular carcinoma (HCC) and the relationship between IGFBP-3 expression and the transforming growth factor-beta (TGF-beta) and/or retinoblastoma (Rb) signaling pathways. In situ hybridization revealed IGFBP-3 transcripts in cancer cells in 6 of 57 (10%) HCCs, including moderately and poorly differentiated HCCs with intrahepatic metastasis. In contrast, all lung metastatic nodules of 4 HCCs showed IGFBP-3 transcripts in cancer cells. The cDNA microarray showed that genes for the TGF-beta pathway and Rb were up regulated in IGFBP-3-expressing HCCs. In 6 HCCs presenting IGFBP-3, immunohistochemical analyses showed abnormalities in the TGF-beta and/or Rb pathways; the loss of phosphorylated-Smad2 was observed in 2, and overexpression of phosphorylated-Rb was observed in the remaining 4 HCCs. The present study suggests that IGFBP-3 mediates growth suppression signals via the TGF-beta and/or Rb pathways in HCC. PMID- 16211217 TI - Isolation and characterization of an N-linked oligosaccharide that is increased in glioblastoma tissue and cell lines. AB - We have isolated and characterized N-linked oligo-saccharides that are significantly increased in glioblastoma tissue and cell lines. The structures of N-linked oligosaccharides present in 3 human normal brain tissues, 15 patients with glio-blastoma and 3 glioma cell lines were analyzed by partially automated technique for the isolation and fluorescent labeling of N-linked sugar chains from glycoproteins. Characterization of the sugar chains was achieved with the use of a combination of HPLC columns and a highly sensitive fluorescence detector at femtomole levels. By collecting peaks which accounted for 0.1% or more, sixteen different oligosaccharide structures were characterized from glioblastoma tissue and cell lines. The 16 oligosaccharide structures accounted for 48.9% of the total N-linked oligosaccharides present in glioblastoma tissue. The major components of total oligosaccharides were similar to those of normal brain tissue. The amount of a biantennary bigalactosylated structure with one core fucosylation (A2G2F) was present in increased levels in glioblastoma tissue (mean = 2.90%) and glioma cell lines (mean = 5.60%), while being less than 0.1% in normal brain tissue. Expression of highly branched tetra-antennary N-glycans that are usually detected in lungs or hepatocellular cancer was not observed. Tissue glioma cells and cultured cells also displayed strong LCA-lectin binding, which binds to sugar chains with core fucose (including A2G2F), while normal brain tissue did not. Moreover, LCA lectin inhibited proliferation of glioma cells through induction of apoptosis. A2G2F on glioma specimens may provide a novel marker and target for the diagnosis and treatment of glioblastoma, respectively. PMID- 16211218 TI - Expression of CD29 on lymphoma cells and/or CD36 on microvascular endothels correlates with high serum LDH level in diffuse large B-cell lymphomas (DLBCLs) and is frequent in de novo CD5-positive DLBCLs. AB - Our previous study demonstrated that CD29 and CD36 mRNAs were overexpressed in de novo CD5-positive diffuse large B-cell lymphomas (CD5+ DLBCLs) compared with CD5 negative (CD5-) DLBCLs. CD29 was expressed on lymphoma cells, and CD36 on microvascular endothels. In the present study, we analyzed the clinicopathological significance of CD29 and microvascular CD36 (mvCD36) expression in many cases of DLBCLs. CD29 and CD36 expression was examined by immunohistochemistry on frozen sections of 159 specimens, consisting of 120 cases of CD5-DLBCL, 14 cases of de novo CD5+ DLBCL, 11 cases of follicular lymphoma, 8 cases of mantle cell lymphoma, and 6 cases of reactive lymphoid hyperplasia. Three cases of CD5- intravascular large B-cell lymphomas (IVLs) with hepatic intrasinusoidal infiltration were also examined. CD29+ and/or mvCD36+ DLBCL cases had significantly higher serum LDH levels compared with CD29- mvCD36- DLBCL cases. The number of CD29+ and mvCD36+ cases, and the number of CD29+ or mvCD36+ cases, were significantly higher in de novo CD5+ DLBCLs than in CD5- DLBCLs. The frequencies of CD29 and mvCD36 expression were low in follicular lymphomas, and mvCD36 was negative in reactive lymphoid hyperplasias. Liver biopsies of three IVL cases showed that CD29 was positive on lymphoma cells, and, that CD36 was expressed on hepatic sinusoids. The present study supports the notion that CD29 and/or mvCD36 expression manifests the aggressive feature of a subset of DLBCLs because of its correlation with high serum LDH levels. Furthermore, CD29 and mvCD36 expression should be evaluated in relation to the intravascular growth pattern of lymphoma cells. PMID- 16211219 TI - Enhancement of the caspase-independent apoptotic sensitivity of pancreatic cancer cells by DHMEQ, an NF-kappaB inhibitor. AB - The effects of the nuclear factor (NF)-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), combined with tumor necrosis factor (TNF) alpha were evaluated in PK-8 pancreatic cancer cells. NF-kappaB was activated by TNF-alpha; however, the administration of DHMEQ abrogated its transcriptional activity. The addition of DHMEQ to TNF-alpha markedly induced apoptosis in PK-8 cells with down-regulation of anti-apoptotic c-FLIP and survivin. Combined treatment significantly suppressed cell viability in vitro, and the anti-tumor effect of DHMEQ was also significant in vivo. We investigated the apoptosis signaling pathway involved in these cell killing effects. Truncated Bid was produced by activated caspase-8, and the subsequent depolarization of the mitochondrial membrane potential (Delta Psi m) peaked at 6 h. Then, the activity of caspase-3 was up-regulated 8-fold. Z-VAD-fmk (a pan-caspase inhibitor) perfectly inhibited the up-regulation of caspase-3 but failed to reverse the cell viability. The above findings indicated that the growth inhibitory effect of combined treatment largely depended on mitochondria-associated caspase independent apoptosis. The intracellular behavior of apoptosis-inducing factor (AIF) following depolarization of Delta Psi m suggested that AIF executed such a caspase-independent apoptosis. Interestingly, caspase-dependent apoptosis appeared within 6 h, whereas the caspase-independent apoptosis lagged. Thus, the addition of DHMEQ to TNF-alpha was capable of inducing caspase-independent apoptosis in pancreatic cancer cells. Once caspase-independent apoptosis was induced, the apoptosis demonstrated powerful cytotoxicity. Therefore, DHMEQ in combination with TNF-alpha may be a promising treatment for pancreatic cancer. PMID- 16211220 TI - Expression of human telomerase reverse transcriptase gene and protein, and of estrogen and progesterone receptors, in breast tumors: preliminary data from neo adjuvant chemotherapy. AB - Human telomerase reverse transcriptase (hTERT), the catalytic subunit of telomerase, is very closely associated with telomerase activity. Telomerase has been implicated in cellular immortalization and carcinogenesis. In situ detection of hTERT will aid in determining the localization of telomerase-positive cells. The aim of this study was to detect expression of hTERT mRNA, hTERT protein, estrogen receptor (ER) and progesterone receptor (PR) in paraffin-embedded breast tissue samples and to investigate the relationship between hTERT expression and various clinicopathological parameters in breast tumorigenesis. We used in situ hybridization (ISH) to examine hTERT gene expression, and immunohistochemistry (IHC) to examine expression of hTERT protein, ER and PR, in breast tissues including 64 adenocarcinomas, 2 phyllode tumors and their adjacent normal breast tissues. hTERT gene expression was detected by ISH in 56 (88%) carcinomas, but in neither of the 2 phyllode tumors. hTERT protein expression was detected by IHC in 52 (81%) carcinomas, but in neither of the 2 phyllode tumors. Moreover, ER and PR were expressed in 42 (66%) and 42 (66%) carcinomas, respectively, and in neither of the 2 phyllode tumors. In 4 cases of breast carcinoma that strongly expressed hTERT gene and protein before treatment, neo-adjuvant chemotherapy led to disappearance of gene and protein expression in all cases. There was a strong correlation between detection of hTERT gene expression by ISH and of hTERT protein by ICH in tissue specimens from breast tumors. These results suggest that detection of hTERT protein by ICH can be used to distinguish breast cancers as a potential diagnostic and therapeutic marker. PMID- 16211221 TI - Cell-cycle regulators, bcl-2 and NF-kappaB in Epstein-Barr virus-positive gastric carcinomas. AB - The mechanism by which Epstein-Barr virus (EBV) contributes to the carcinogenesis of gastric mucosa remains unanswered. In this study, the role of cell-cycle regulators (p53, p21, p27, p16, cyclin D1, Rb), bcl-2 and NF-kappaB p65 (Rel A) was evaluated. Immunohistochemistry for these proteins was performed in EBV positive (n=55) and EBV-negative gastric carcinomas (n=72). The bcl-2 protein by western blot and EBV transcripts using RT-PCR were studied in cell lines. The p27 loss, p16 loss, cyclin D1 expression and NF-kappaB nuclear positivity were more frequent in EBV-positive gastric carcinomas than those in EBV-negative gastric carcinomas, while p53 overexpression seldom occurred in EBV-positive carcinomas (p<0.001). EBV-positive gastric carcinoma showed unique p53 immunostaining (heterogeneous, weak to moderate, focal staining), and rare bcl-2 positivity (1 case). Western blot showed bcl-2 to be irrespective of EBV status in stomach cancer cell lines. However, bcl-2 was highly expressed in EBV-positive lymphoma or EBV-positive lymphoblastoid cell lines. The BARF1 transcript was confirmed in both EBV-positive stomach cancer and EBV-positive lymphoma, suggesting tissue type-specific bcl-2 activation by BARF1. The pathological tumor stage was the only independent prognostic factor. A small size of tumor, p16 preservation and NF-kappaB nuclear positivity were associated with a good prognosis in univariate analysis (p<0.05). p27, p16, cyclin D1 and NF-kappaB may be associated with oncogenesis in EBV-positive gastric carcinomas. EBV-positive gastric carcinomas showed infrequent p53 overexpression, wild-type p53 stabilization and rare bcl-2 involvement. The characteristic expression of proteins may relate to both EBV and tissue type. PMID- 16211222 TI - Cellular retinoic acid-binding protein 2 is down-regulated in prostate cancer. AB - Prostate cancer is among the most frequent tumours in industrialized nations and many questions remain open concerning the molecular events underlying its development and progression. In the present study we have combined cDNA array hybridization to laser-assisted microdissection (LAM) in order to investigate differences in gene expression between epithelial and stromal cells of prostate cancer and normal peripheral prostate tissue. Results have been verified for selected candidate genes by quantitative real-time RT-PCR. Using this approach and immunohistochemistry we could demonstrate a down-regulation of cellular retinoic acid binding protein 2 (CRABP2) mRNA and protein in carcinoma cells compared to normal glandular cells. CRABP2 is a main regulator of anti carcinogenic activities of retinoic acid and may become a novel diagnostic marker and experimental therapeutic tool for prostate cancer. In addition, results of cDNA array hybridization suggest an up-regulation of 34 further genes and a down regulation of 6 genes in cancer tissues compared to normal peripheral prostate tissues. Several of these genes have already been reported to be associated with carcinogenesis in organs such as the prostate. PMID- 16211223 TI - Neuropilin-1 and VEGF correlate with somatostatin expression and microvessel density in ovarian tumours. AB - Vascular endothelial growth factor (VEGF), is an angiogenic growth factor, expressed more highly in malignant than benign ovarian tumours. Neuropilin-1, which can act as a VEGF receptor has been shown to be associated with tumour angiogenesis in some cancer systems. Somatostatin (SST), a potentially anti angiogenic factor, acts via somatostatin receptors that are expressed in ovarian cancer. We used immunohistochemistry to demonstrate expression of Neuropilin-1 in 63 malignant and 35 benign ovarian tumours and compared it to VEGF, Flt, Flk, SST expression and tumour microvessel density (MVD). Neuropilin-1 was expressed in 34/63 malignant and 22/35 benign lesions. VEGF, Flt, Flk and SST were expressed more highly in the epithelium of malignant and the vessels of benign lesions. VEGF expression correlated with SST expression in the epithelium (p<0.001) and the vessels (p<0.001), this co-expression was confirmed by dual immunostaining. The MVD for malignant lesions was higher than benign (p<0.001) and positively correlated to epithelial VEGF expression (p=0.001) and negatively correlated to vascular VEGF expression (p=0.025). These results show that Neuropilin-1 is expressed in ovarian tumours and also show that VEGF and SST are co-expressed in the same tissue compartments raising the intriguing possibility that SST may be important in angiogenesis in ovarian cancer. PMID- 16211224 TI - Expression of MUC1, MUC2 and oligosaccharide epitopes in breast cancer: prognostic significance of a sialylated MUC1 epitope. AB - MUC1 represents a promising marker in breast cancer. However, due to the structural complexity of the MUC1 glycoprotein, multiple epitopes can be detected by monoclonal antibodies. This fact may be responsible for the contradictory results of previous investigations regarding the clinical and prognostic relevance of MUC1 expression in breast cancer. Therefore, we tried to evaluate the role of different glycosylated and non-glycoslyated MUC1 epitopes as well as other mucin-associated peptides (MUC2) and carbohydrates (Thomsen-Friedenreich antigen, sialyl-Lewisa, sialyl-Lewisx) as predictors of the clinical course and prognosis in mammary carcinomas. An immunohistochemical study applying numerous monoclonal antibodies (mabs) was performed to characterize the expression of a selected panel of MUC1 epitopes, and of MUC2, Thomsen-Friedenreich antigen, sialyl-Lewisa, and sialyl-Lewisx in a series of 140 patients with breast cancer. The results were correlated with clinicopathological variables as well as overall survival. Generally, more than 90% of the mammary cancers, were strongly stained with the MUC1-specific mabs. Especially ductal and lobular carcinomas were strongly MUC1- and sialyl Lewisa-positive, whereas MUC2 binding was significantly elevated in mucinous neoplasms. Associations between the immunoreactivity of any mab under study and tumor progression as reflected by pTNM staging could not be observed. However, expression of the sialylated MUC1 epitope detected by mab MY1.E12 was revealed as a favourable independent prognostic factor. These results confirm that MUC1 is generally strongly expressed in mammary carcinomas. As an exception, mucinous carcinomas are significantly less MUC1 reactive, but strongly express MUC2. Our data suggest that only the presence of a sialylated short-chain MUC1 glycoform is associated with a better prognosis, whereas the other molecules under study are not correlated with the course of disease and survival probability. PMID- 16211225 TI - Synthetic microRNA and double-stranded RNA targeting the 3'-untranslated region of HER-2/neu mRNA inhibit HER-2 protein expression in ovarian cancer cells. AB - MicroRNA (miRNA) are a class of non-coding RNAs found both in normal tissues and cancer cells. The natural miRNA, which target cancer genes for transcriptional repression are still unknown. Approaches for synthetic miRNA design targeting cancer genes have not yet been established. We designed miRNAs targeting the 3' UTR (nt 4350-4372) of HER-2 proto-oncogene. One miRNA (miR-14U) was designed by introducing a mutation in the nt 14 counting from the 5' end of anti-sense RNA. Two others (miR4350-10GGA and miR4350-11AAGCU) were designed by introducing either loop-forming nucleotides in position 10 or a part of the complementary sequence of the Brd-box consensus sequence, in position 11. miR4350-10GGA was more effective than the anti-sense strand in decreasing the numbers of ovarian tumor SKOV3 cells, which over-expressed HER-2 protein. Its inhibitory effects were lower than that of corresponding double-stranded (ds) RNA4350-4372. Inhibition of HER-2 expression mediated by miRNAs was higher in cells expressing higher levels of HER-2 protein than in cells expressing lower levels of HER-2 protein. This is the first demonstration of inhibition of expression of a constitutively over-expressed tumor protein by designed synthetic miRNA and its cor-responding dsRNA targeting 3' untranslated regions in mRNA. Our results also show that measuring the effects of miRNA and dsRNA in pooled populations of tumor cells, which express various levels of target oncogenes, may reflect the survival responses of siRNA resistant tumors rather than the growth inhibitory responses of siRNA-sensitive tumors. PMID- 16211226 TI - How to best classify breast cancer: conventional and novel classifications (review). AB - Breast cancer is a complex disease and different classifications, mostly based on clinical and pathological features, have been used for guiding the management of patients. Most of them fail to reflect breast cancer heterogeneity, which could be the reason why the treatment fails in approximately 30% of cases. Emerging molecular studies based on gene expression profiling using DNA microarrays have defined new molecular subtypes of breast cancer associated with the cell-of origin distinction. Thus, breast cancer has been divided into five subgroups with distinct biological features and clinical outcomes. We have tried here to confront the conventional existing classifications with this new molecular taxonomy. It is likely that using all types of classification together will help in the management of breast cancer. PMID- 16211227 TI - OGG1 Cys326 variant, allelic imbalance of chromosome band 3p25.3 and TP53 mutations in ovarian cancer. AB - A Ser326Cys amino acid variant encoded by OGG1 that removes the highly mutagenic 8-oxo-7,8-dihydroguanine adducts resulting from the oxidative damage produced by reactive oxygen species, has been suggested to reduce activity of the enzyme and result in an increase in G:C --> T:A transversions. Carriers homozygous for the Cys326 variant have been associated with increased risk in some cancers. In this study, we tested the association of this polymorphism with epithelial ovarian cancer (EOC) (n=91 cases) in comparison with women unaffected by the disease (n=57). Although the genotype encoding the homozygous Cys326 variant was found more often in cases (11%) than controls (9%), this was not statistically significant (p=0.297). As OGG1 at chromosome band 3p25.3 overlaps a region exhibiting allelic imbalance (AI) in EOC, we also examined the frequency of AI of the OGG1 locus and its association with somatic mutations in the TP53 tumor suppressor gene, which is frequently mutated in EOC. The frequency of AI and TP53 mutations in EOC was 32 and 37%, respectively. Although the proportion of EOC cases harboring the genotype homozygous for Cys326 was greater in TP53 mutation positive cases (5 of 34, 15%) than mutation-negative cases (5 of 57, 9%), the distribution of genotypes was not significantly different (p=0.289). Four of the six heterozygous Ser326/Cys326 samples exhibiting AI, displayed loss of the Ser326 variant. About 16% of samples exhibited both AI of the OGG1 locus and a TP53 mutation. Proportionately more EOC samples harbored a mutation in the group of samples exhibiting AI (15 of 29, 52%) in comparison to the group of samples with no AI (19 of 62, 31%) (p=0.036). Although the OGG1 Cys326 variant appears to play a minor role in conferring increased risk of ovarian cancer, the observed AI of the OGG1 locus in association with TP53 somatic mutations warrants further investigation in this disease. PMID- 16211229 TI - Overexpression of G protein-coupled receptors in cancer cells: involvement in tumor progression. AB - G protein-coupled receptors (GPCRs) play important roles in a variety of biological and pathological processes. They are considered among the most desirable targets for drug development. Recent studies have demonstrated that many GPCRs, such as endothelin receptors, chemokine receptors and lysophosphatidic acid receptors have been implicated in the tumorigenesis and metastasis of multiple human cancers. In this study, we conducted an in silico analysis of GPCR gene expression in primary human tumors by analyzing some publicly available gene expression profiling data. Statistical analysis was performed on eight microarray data sets of non-small cell lung cancer, breast cancer, prostate cancer, melanoma, gastric cancer and diffused large B cell lymphoma to identify GPCRs that are up-regulated in primary or metastatic cancer cells. Our analysis has demonstrated overexpression of several GPCRs in primary tumor cells, including chemokine receptors and protease-activated receptors that were shown to be important for tumorigenesis by previous studies. In addition, we have uncovered several GPCRs, such as neuropeptide receptors, adenosine A2B receptor, P2Y purinoceptor, calcium-sensing receptor and metabotropic glutamate receptors, that are expressed at a significantly higher level in some cancer tissue and may play a role in cancer progression. Analysis of cancer samples in different disease stages also suggests that some GPCRs, such as endothelin receptor A, may be involved in early tumor progression and others, such as CXCR4, may play a critical role in tumor invasion and metastasis. The present study demonstrates the value of publicly available microarray data as a resource to gain more understanding of cancer biology, to validate previous findings from in vitro experiments, and to identify potential novel anticancer targets and biomarkers. PMID- 16211228 TI - EIPL (extensive intraoperative peritoneal lavage) therapy significantly reduces peritoneal recurrence after pancreatectomy in patients with pancreatic cancer. AB - One standing problem in the therapy for pancreatic cancer is peritoneal recurrence after pancreatectomy. We have established EIPL (extensive intra operative peritoneal lavage) therapy as a very simple and non-aggressive prophylactic treatment for peritoneal dissemination of gastric cancer patients with peritoneal free cancer cells. In this study, the impact of EIPL therapy in preventing peritoneal recurrence in patients with pancreatic cancer after a curative surgical operation is analyzed. This study was based on 39 consecutive patients with invasive ductal adenocarcinoma of the pancreas who underwent curative surgical treatment. The patients were divided into two groups: the non EIPL group, patients without EIPL therapy (from 1995 to 2000) (n=24); and the EIPL group, patients with EIPL therapy (from 2001 to 2003) (n=15). Clinicopathological findings, recurrence patterns and cancer-free survival were compared between the two groups. In addition, to identify free cancer cells during the operation, both carcinoembryonic antigen (CEA)- and cytokeratin 20 (CK20)-specific intraoperative real-time RT-PCR was performed immediately after laparotomy, lymph node dissection and removal of the pancreas in six patients with curative pancreatic cancer. Peritoneal, hepatic, lymphatic, local, and extra abdominal recurrence rates in the EIPL and non-EIPL groups were 6.7, 40.0, 26.7, 13.3 and 13.3%, and 45.8, 50.0, 20.8, 29.2 and 20.8%, respectively. Among these recurrence patterns, the peritoneal recurrence rate of the EIPL group was significantly lower than that of the non-EIPL (P=0.013). Real-time RT-PCR analysis identified intra-peritoneal free cancer cells in 0, 33 and 67% of patients immediately after laparotomy, lymph node dissection and removal of the pancreatic cancer, respectively. After EIPL therapy, however, no cancer cell was detected in the peritoneal cavity. EIPL therapy was the independent negative risk factor for peritoneal recurrence (P=0.044), and the cancer-free 2-year survival of the EIPL group showed improvement compared with that of the non-EIPL group and was near statistical significance (P=0.097). EIPL therapy significantly prevented peritoneal recurrence after curative operation of pancreatic cancer, and it may contribute to reducing the mortality rate of this aggressive disease. Continued investigation of this promising treatment regimen is warranted. PMID- 16211230 TI - Pharmacological breast cancer therapy (review). AB - Breast cancer ranks as the second most common cause of cancer death among women in the United States. Anticancer agents are an important component of breast cancer therapy. Drugs frequently used to treat breast cancer include methotrexate, 5-fluorouracil (5-FU), cyclophosphamide, anthracyclines, taxanes, trastuzumab, tamoxifen, and aromatase inhibitors. These agents inhibit breast cancer progression by a variety of different mechanisms. This review describes each of these drugs and the varying effects each of them have upon breast cancer cells. PMID- 16211231 TI - Tumor-associated serpin, squamous cell carcinoma antigen stimulates matrix metalloproteinase-9 production in cervical squamous cell carcinoma cell lines. AB - Squamous cell carcinoma antigen (SCCA) is clinically used as a tumor marker for patients with squamous cell carcinoma of various organs. SCCA1 and its highly homologous molecule, SCCA2, belong to the serine proteinase inhibitors (serpins) family, suggesting that these proteins may be involved in the malignant behavior of squamous cell carcinoma cells. The aim of this study is to functionally characterize these tumor-associated serpins regarding the potential to influence the production of matrix metalloproteinases (MMPs), which play a key role in tumor cell invasion and metastasis. Cervical squamous cell carcinoma cell lines, CaSki cells and SKG-IIIa cells were incubated with SCCA1 or SCCA2 and MMP production was analyzed by gelatin zymography. Both SCCA1 and SCCA2 significantly increased production of proMMP-9, but not proMMP-2. These stimulatory effects were still observed when cells were treated with SCCA mutants lacking the proteinase inhibitory activity of serpins. Furthermore, treatments with various forms of SCCAs, which are generated by interacting with their target proteinases, diminished the stimulatory effect of SCCAs, implying the importance of the conformational structure of SCCAs in the stimulatory effects of SCCAs on proMMP-9 production. In addition, in vitro invasion assay showed that SCCA1 and SCCA2 significantly promoted the activity of cell invasion. It is concluded that SCCAs can alter the invasive phenotype of cervical squamous cell carcinoma cells, probably by stimulating proMMP-9 production, and that intact conformational structure of SCCAs, but not proteinase inhibitory activity of serpins, is required for its stimulatory activity on proMMP-9 production. PMID- 16211232 TI - An aminoacridine derivative for radionuclide therapy: DNA binding properties studied in a novel cell-free in vitro assay. AB - Radiolabelled DNA-binding compounds can be used to increase the efficiency of radionuclide cancer therapy of disseminated disease. In this work, the aminoacridine compound N-[3-(acridine-9-ylamino)-propyl]-3-iodobenzamide (A3) labelled with the Auger-emitting nuclide 125I using Chloramine-T was studied. Optimal labelling conditions of 125I-A3 were investigated and the interaction with DNA was studied using a novel cell-free in vitro assay with naked human genomic DNA in agarose plugs. This novel assay showed to be simple and reliable. The results verify that 125I-A3 specifically binds DNA with low dissociation and is potent in causing double-strand breaks, yielding 1.0-1.4 breaks per decay. In conclusion, 125I-A3 is a most suitable DNA-binding compound for future therapeutic studies of Auger-electron emitters like 125I. PMID- 16211234 TI - Identification of human papillomavirus 16-E6 protein-derived peptides with the potential to generate cytotoxic T-lymphocytes toward human leukocyte antigen-A24+ cervical cancer. AB - Human papillomavirus 16 (HPV16)-E6 and -E7 proteins are considered to be appropriate targets in specific immunotherapy for cervical cancer. In this study, we attempted to identify epitope peptides from the HPV16-E6 protein that have the potential to generate cytotoxic T-lymphocytes (CTLs) toward human leukocyte antigen (HLA)-A24+ cervical cancer. Two HPV16-E6 peptides at positions 75-83 and 91-100 effectively induced peptide-specific CTLs from peripheral blood mononuclear cells of HLA-A24+ cervical cancer patients. These HPV16-E6 peptide induced CTLs showed cytotoxicity against HLA-A24+ and HPV16-E6 protein-expressing cervical cancer cells. Experiments with blocking antibodies and cold inhibition targets revealed that the cytotoxicity was mainly dependent on peptide-specific and CD8+ T cells. In addition, based on our observation that induction of immunoglobulin G (IgG) reactive to administered CTL-directed peptides is correlated with clinical responses, we attempted to detect IgG reactive to HPV16 E6 peptides in the plasma of cancer patients. As a result, IgGs reactive to the HPV16-E691-100 peptide were detected in 4 of 12 cervical cancer patients. These results indicate that these HPV16-E6-derived peptides are good candidates in peptide-based immunotherapy for HLA-A24+ cervical cancer patients. PMID- 16211233 TI - Serum amyloid A as a tumor marker in sera of nude mice with orthotopic human pancreatic cancer and in plasma of patients with pancreatic cancer. AB - We screened an orthotopic nude mouse model of human pancreatic cancer for candidate serum biomarkers and examined their presence in the plasma of pancreatic cancer patients. Nude mice were injected in the pancreas with L3.9pl human pancreatic cancer cells. One week later, the mice were randomized into 4 treatment groups: i) control, saline; ii) oral STI 571; iii) intraperitoneal gemcitabine; and iv) STI 571 and gemcitabine. After 1, 2, and 3 weeks of treatment, sera and tumors were collected from mice in each group as well as uninjected mice. All sera were analyzed by surface enhanced laser desorption ionization mass spectrometry using ProteinChip technology. Protein profiles were analyzed with the Biomarker Wizard software package. The concentration of candidate proteins was evaluated in mouse sera and plasma from 135 pancreatic cancer patients, 7 pancreatitis patients, and 113 healthy volunteers. The combination therapy inhibited tumor growth. A 11.7-kDa protein peak correlating with tumor weight was purified by gel filtration, separated by SDS-PAGE, and identified as mouse serum amyloid A (SAA) by amino acid sequencing and public database searches. The expression of SAA in mouse sera was confirmed by Western blotting and correlated with tumor weight. The level of SAA in plasma of pancreatic cancer patients correlated with clinical stage and was significantly higher than in normal volunteers (mean value: 180.1 microg/ml vs 27.9 microg/ml: P<0.01) or pancreatitis patients. For SAA used as a single tumor marker with a cut-off of 75 microg/ml, the sensitivity for pancreatic cancer was 96.5% and specificity was 31.9%. Our search for specific marker proteins to identify pancreatic cancer was unsuccessful. Although SAA is not specific for pancreatic cancer and not sensitive enough to detect stage I patients, it may be a candidate biomarker for detecting and monitoring the progressive growth of pancreatic cancer. PMID- 16211235 TI - The role of human prolactin and its antagonist, G129R, in mammary gland development and DMBA-initiated tumorigenesis in transgenic mice. AB - Human prolactin (hPRL) has been implicated to have a pathological role in breast cancer and play a critical role in mammary gland development. The hPRL antagonist, G129R, has been shown to induce breast cancer cell apoptosis. 9,10 Dimethyl-1,2-benzanthracene (DMBA), a potent mammary gland carcinogen, induces hormone responsive mammary tumor formation in rodents. To investigate the effects of hPRL and its counterpart, G129R, on mammary gland development and tumorigenesis, transgenic mice that express hPRL or G129R under the regulation of the metallothionein (Mt) promoter were generated. Mammary glands from virgin female transgenic mice at the ages of 12, 24, and 36 weeks were used to compare the effect of hPRL and G129R in various developmental stages. Mammary gland whole mount comparisons between transgenic mice and their littermates revealed a significant increase in ductal branching and lobular bud formation in hPRL transgenic mice; whereas a drastic decrease in ductal branching and lobular bud formation was observed in the mammary glands of G129R transgenic mice. In addition, total RNA isolated from the mammary glands of transgenic mice at the three different ages was analyzed on Affymetrix GeneChip Mouse Expression 430A chips (MOE430A). Microarray data revealed alteration to the gene expression levels, greatest at 12 and 36 weeks. Furthermore, hPRL and G129R transgenic mice, as well as their littermates, were treated with multiple doses of DMBA and the rate of mammary tumor formation and survival were compared. The tumor rates in the G129R transgenic mice were significantly reduced (18% at 28 weeks) as compared to that of either NTG (39%) or hPRL (40%). On the other hand, the tumor appearance is significantly earlier in the PRL transgenic group as compared to that of controls. Taken together, the data further confirmed the inhibitory effects of G129R in mammary gland development, which translates to a resistance to DMBA-initiated breast tumorigenesis. PMID- 16211236 TI - Indole-3-carbinol inhibits the growth of human colon carcinoma cells but enhances the tumor multiplicity and volume of azoxymethane-induced rat colon carcinogenesis. AB - Indole-3-carbinol (I3C) is a naturally occurring phytochemical which exerts a broad range of biological activities. The purpose of this study was to examine the effects of I3C on colon carcinogenesis, cell proliferation, cell-cycle progression and apoptosis, and on the levels of expression of several cell-cycle control molecules. We used a long-term rat model by using azoxymethane (AOM) to induce tumors (adenomas and adenocarcinomas) in the colon. In the present study, we found that after AOM injection, the treatment of male F344 rats with 0.01 and 0.05% I3C caused a significant increase in the tumor multiplicity of adenocarcinomas by 2.2- (P<0.05 for 0.01% I3C) and 2.1-fold (P<0.0002 for 0.05% I3C) respectively, when compared to the control rats. In addition, the tumor multiplicity of adenoma plus adenocarcinoma and the volume of adenocarcinoma were also increased by 2.0- (P<0.00001) and 2.1-fold (P<0.05) respectively, compared to the control. I3C significantly increased the proliferating cell nuclear antigen labeling index (PCNA LI) (P<0.008) and decreased the apoptotic index (P<0.05) of the colon adenocarcinoma. In contrast, in HCT 116 and HT29 human colon carcinoma cells, I3C inhibited growth and induced G1-phase cell-cycle arrest and apoptosis. Furthermore, I3C caused approximately a 2- to 4-fold increase in the cellular levels of p27KIP1 and p21CIP1 mRNA. These results suggest that I3C inhibits the growth of human colon carcinoma cells, at least in part, by inducing p27KIP1 and p21CIP1-mediated G1 cell-cycle arrest but dietary I3C promotes AOM-induced rat colon carcinogenesis by inhibiting the apoptosis of colon tumors. Therefore, the present study may provide further evidence for the ambivalent modulatory activity of I3C and this information may be useful when including I3C in cancer chemoprevention and/or extensive clinical therapy trials. PMID- 16211237 TI - HMG-CoA reductase inhibitors (statins) as anticancer drugs (review). AB - Apart from their lipid lowering activity, HMG-CoA reductase inhibitors (statins) impair numerous cellular functions associated with metastasis, e.g. gene expression, angiogenesis, cell adhesion, cell motility and invasiveness. Furthermore, statins have impact on apoptotic cell death and modulate cellular susceptibility to cell killing by anticancer drugs and ionizing radiation. Part of the effects provoked by statins are due to the inhibition of the prenylation of low molecular weight GTPases, in particular Ras and Rho, which play key roles in signaling evoked by stimulation of cell surface receptors. C-terminal lipid modification of Ras/Rho GTPases is essential for their correct intracellular localization and function. By depletion of the cellular pool of isoprene precursor molecules, statins reduce the level of membrane-bound active Ras/Rho proteins, thereby impairing corresponding functions. Since broad clinical experience already exists for statins, their incorporation into established tumor therapeutic regimens would be realizable in a rather short period of time. Here, data available at present arguing for the usefulness of statins in anticancer therapy are summarized and discussed. PMID- 16211238 TI - The tuberous sclerosis genes, TSC1 and TSC2, trigger different gene expression responses. AB - Tuberous sclerosis (TSC) is an autosomal dominant tumor suppressor gene syndrome affecting about 1 in 6000 individuals. It is characterized by mental retardation and epilepsy. A variety of tumors characteristically occur in different organs of TSC patients. The genes, TSC1 on chromosome 9q34, encoding hamartin, and TSC2 on chromosome 16p13.3, encoding tuberin are responsible for TSC. Hamartin and tuberin form a complex providing a tentative explanation for the similar disease phenotype in TSC patients with mutations in either of these genes. Besides overlap in many features of patients with TSC1 and TSC2 mutations, data accumulated provide evidence for specific clinical differences. Here, we performed microarray analyses of the gene expression response to overexpressed TSC1 or TSC2 in HeLa cells. Out of 2400 analysed genes we found 115 genes to be up-regulated > or =2-fold upon ectopic TSC1 overexpression and 284 genes to be up regulated > or =2-fold via TSC2. Only 34 of these genes were up-regulated by both, TSC1 and TSC2. Whereas only 7 genes were down-regulated > or =2-fold via TSC1, ectopic TSC2 triggered a > or =2-fold down-regulation of 113 genes. Only 3 of these genes were down-regulated by TSC1 and TSC2. This study provides new insights into the cellular roles of TSC proteins and promotes discussion on whether separable functions of these proteins might be associated with the clinical differences of TSC1- and TSC2-associated disease. PMID- 16211239 TI - Expression of fibrinogen E-fragment and fibrin E-fragment is inhibited in the human infiltrating ductal carcinoma of the breast: the two-dimensional electrophoresis and MALDI-TOF-mass spectrometry analyses. AB - In the present study, total proteins from a tissue of an infiltrating ductal carcinoma of the breast (IDCA) were compared by the two-dimensional electrophoresis (2D-PAGE) to proteins from an adjacent non-neoplastic breast tissue. Analysis of multiple gels for each sample identified nine proteins present in the tumor sample that were less present in the matched normal adjacent breast tissue and four proteins present at higher levels in the normal tissue. The altered proteins were identified by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and search in protein databases. Protein disulfide isomerase, BiP protein, calreticulin, cathepsin D, inorganic pyrophosphatase, vimentin, apolipoprotein A1 precursor, tropomyosin 4 and beta5-tubulin were identified as being significantly over expressed in the IDCA with regard to the normal tissue. The expression of fibrinogen E-fragment (known as anti-angiogenic factor) as well as of fibrin E, Pro2619 and actinG1 was found to be inhibited in the tumor sample. The identified proteins might play an important role during malignant transformation, breast cancer progression, and angiogenesis as well as in cellular signaling. This study demonstrates quantitative and qualitative changes in protein abundance between IDCA and normal tissue. The identification of these differentially expressed proteins could lead to a better understanding of the molecular events linked to breast cancer progression. PMID- 16211240 TI - Overexpression of Midkine in malignant peripheral nerve sheath tumor cells inhibits apoptosis and increases angiogenic potency. AB - Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive soft tissue tumors arising sporadically although more frequently in patients with Neurofibromatosis type 1. Prognosis remains dismal as chemo- and radiotherapy have not been shown to be successful. The heparin-binding growth factor, Midkine (MK), is implicated in the tumorigenesis of benign and plexiform neurofibromas, and thereof arising MPNSTs. MK is mitogenic, anti-apoptotic, angiogenic and can promote tumorigenicity in several cell types. Thus, we investigated the role of MK in malignant biology and tumorigenicity in MPNSTs by stable transfection into MPNST cell lines. Overexpression of MK in the MPNST cell line, S462, increased cell viability and protected cells from apoptosis under serum deprivation, but did not induce proliferation. In addition, MK-transfected S462 cells were partially protected from vincristine-induced cell death. Conditioned medium of MK transfected S462 cells was a potent mitogen for human umbilical venous endothelial cells. Furthermore, MK overexpression in S462 cells was accompanied by higher levels of VEGF mRNA. Yet, stable overexpression of MK in S462 as well as in ST88-14 cells was not sufficient to promote xenograft tumor growth in nude mice. However, increasing survival and enhanced angiogenic potency of MK transfected S462 cells highlight the importance of developing specific inhibitors for MK as part of new therapeutic concepts against MPNSTs. PMID- 16211241 TI - Targeted inhibition of transient activation of the EGFR-mediated cell survival pathway enhances paclitaxel-induced ovarian cancer cell death. AB - Emerging studies have suggested that transient activation of the cell survival pathway may be the strategy for cancer cells to fight against chemotherapy and eventually mysteriously evade paclitaxel-induced cell death. Modulation of the EGFR-mediated survival pathway in addition to the utilization of paclitaxel renders a promise of better clinical management. The objective of this study was to understand the molecular mechanism of transient induction of EGFR-mediated cell survival by paclitaxel. We utilized ovarian cancer cell line, Caov3, cells to investigate the effect of paclitaxel on EGFR-mediated MAP kinase and AKT activation, and the expression of survivin. We found that paclitaxel transiently induced EGFR phosphorylation and ERK and AKT activation but not JNK and p38. Paclitaxel-induced ERK and AKT activity was inhibited by the EGFR inhibitor, PD153035; ERK inhibitor, U0126; and PI3 kinase inhibitor, LY294002, respectively. We observed that paclitaxel transiently induced expression of survivin in the early hours of treatment. Paclitaxel-induced survivin expression was inhibited by the EGFR inhibitor, PD153035. Inhibitors of EGFR, ERK and PI3 kinase all enhanced paclitaxel-induced cell death. We conclude that paclitaxel transiently transactivates EGFR, leading to activation of cell survival factors, such as ERK and AKT, and expression of survivin, which are all inclusively accountable for ovarian cancer cell resistance to paclitaxel treatment. A combination of these inhibitors with paclitaxel may be a better option for ovarian cancer treatment. PMID- 16211242 TI - Effects of MEK1 and PI3K inhibitors on allyl-, benzyl- and phenylethyl isothiocyanate-induced G2/M arrest and cell death in Caco-2 cells. AB - Isothiocyanates (ITCs) are potentially important cancer chemopreventive compounds found in cruciferous vegetables. In this study, three ITCs: allyl ITC, benzyl ITC and phenylethyl ITC, induced DNA cell-cycle changes and cell death in undifferentiated Caco-2 cells and their roles in PI3K/Akt and MEK/ERK signaling pathways have been investigated. Flow cytometric analysis was used to measure cell-cycle distribution, expression of mitotic marker (phosphorylated H3 histone), mitochondrial transmembrane potential for the determination of ITC induced apoptosis measured by Annexin V-FITC staining and metabolic conversion of fluorescein diacetate, and quantification of sub-G1 population. Cellular MAPK and phosphorylated MAPK were measured using western blot analysis. All ITCs tested induced G2/M cell-cycle arrest after 24-h treatment, a time- and concentration dependent activation of ERK1/2, dissipation of mitochondrial transmembrane potential and apoptosis. Both PI3K/Akt and MEK/ERK inhibitors, LY294002 and PD98059, attenuated the extent of BITC-induced cell death. Pretreatment of cells with either the PD98059 or LY294002 inhibitor, caused a dose-dependent inhibition of histone H3 (p-H3) phosphorylation. Despite the LY294002 inhibitor having no effect on the proportion of ITC-induced G2/M arrested cells, a significant decrease of p-H3/(G2/M) ratio in both PD98059- and LY294002-treated cells was observed. We suggest that the decrease of mitotic cells was compensated for by an increase of cells in G2 phase. LY294002 and PD98059 affect cell transition from G2 to M phase and from S to G2 phase respectively. These results indicate that isothiocyanates can induce cell cycle-change through multiple signaling pathways and more detailed study is merited to further unravel the chemopreventive and chemotherapeutic mechanisms of ITCs. PMID- 16211243 TI - Deregulation of estrogen induced telomerase activity in tamoxifen-resistant breast cancer cells. AB - Telomerase, a ribonucleoprotein enzyme that functions as a reverse transcriptase, is detected exclusively in immortal cells such as germ cells, stem cells and cancer cells. Telomerase activity is present in almost all human cancers. Telomerase activation is considered to be essential to maintain the integrity of the replicating tumor cell and to establish immortality. Based on this concept antiestrogen should initially regulate estrogen-stimulated telomerase but the enzyme would be expected to be constitutive in tamoxifen-resistant tumor cells. We have studied the estrogen regulation of telomerase in T47D:A18 breast cancer cells with a TRAPEZE Telomerase detection kit. Estradiol significantly increased telomerase activity after a 2-day treatment. Telomerase activity induced by estradiol was up to 10-fold higher within 4 days. Antiestrogens 4 hydroxytamoxifen (4-OHT) and ICI 182,780 were inactive alone and significantly blocked estradiol-stimulated increase in telomerase. These effects were correlated with changes in cell replications and changes in the cell cycle. In contrast, 4-OHT resistant T47D:A18 cells (T47D:A18/4-OHT, cultured in 1 microM 4 OHT for 6 months) grew spontaneously and had no changes in the cell cycle with estrogen treatment. The estrogen receptor (ERalpha) was present and still regulated at an estrogen responsive luciferase reporter gene with estrogen despite the fact that progesterone receptor was not increased in response to estradiol in T47D:A18/4-OHT cells. However, telomerase activity was increased about 40-fold in T47D:A18/4-OHT cells and this was not regulated by ICI 182,780. We conclude that the differential regulation of telomerase gene might be an important transition for tamoxifen resistance in T47D:A18 breast cancer cells. PMID- 16211244 TI - Blockage of the ERK signaling pathway abrogates the SCAP ligand-induced transcriptional activation of the LDL receptor gene in HepG2 cells. AB - Previous studies identified the putative SCAP ligands including compound GW707 as a new class of up-regulators of LDL receptor (LDLR) transcription by activation of the sterol-regulatory element binding proteins (SREBP). These compounds increase LDLR expression in hepatoma cells in vitro and lower plasma LDL-c in hamsters. However, it is unknown, what signaling pathways are utilized by these agents that lead to the activation of LDLR transcription. Here, we report that the ERK signaling cascade is critically involved in GW707-mediated induction of LDLR expression. We show that: a) blocking ERK activation with U0126, the inhibitor of ERK upstream kinase MEK, completely abolishes the inducing effects of GW707 on LDLR promoter activity, LDLR mRNA expression, and DiI-LDL uptake in HepG2 cells; b) treating HepG2 cells with GW707 induces a dose-dependent conversion of SREBP-2 from the 125 kDa precursor form to the 68 kDa activated form and U0126 does not inhibit this cleavage process, but U0126 significantly reduces the total amount of SREBP-2 protein in GW707-treated cells without affecting the expression levels of other proteins involving in SREBP processing; and c) inhibition of ERK signaling pathway has no effects on the promoter activity or mRNA expression of SREBP-2. Collectively, these new findings establish an important role of ERK signaling pathway in SCAP ligand-induced transcription of LDLR and imply that the protein synthesis or turnover rate of SREBP-2 may be regulated by ERK. PMID- 16211245 TI - G protein-coupled APJ receptor signaling induces focal adhesion formation and cell motility. AB - APJ, a G protein-coupled receptor, has an endogenous ligand called apelin. APJ and apelain are highly expressed in the cardiovascular system from embryo to adulthood. It has been shown that apelin elicited the migration of APJ-expressing cells, but details of the receptor signaling have not been identified. To address the signal transduction molecular mechanisms of the apelin/APJ-induced cell motility, we established human embryonic kidney 293T cells stably expressing the mouse APJ (APJ/293T). APJ/293T cells exhibited a specific [(Glp65, Nle75, Tyr77) [125I]]-Apelin13 binding activity (Kd = 4.45 nM). Apelin induced Akt/PKB phosphorylation in APJ/293T cells, but not in the intact 293T cells (-/293T cells). This APJ-dependent activation of Akt/PKB was significantly inhibited by the pretreatment of pertussis toxin (PTx) and a PI3K inhibitor, LY29004. In addition, apelin enhanced focal adhesion kinase (FAK) phosphorylation and increased focal adhesion formation with staining for F-actin in APJ/293T cells. PTx and LY29004 significantly suppressed these responses to apelin. Moreover, we examined the migration activity by using a scratch-test. Apelin strongly accelerated the cell motility in APJ/293T cells, and this activity was abolished by PTx and LY29004. These results indicated that the apelin/APJ signaling coupled with the PTx-sensitive G-protein activates Akt/PKB and FAK proteins through PI3K. PMID- 16211246 TI - The exploration of joint-specific immunoreactions on immunoglobulins G of anti glucose-6-phosphate isomerase antibody-positive patients with rheumatoid arthritis. AB - The pathogenic role of autoantibodies in rheumatoid arthritis (RA) remains elusive. Anti-glucose-6-phosphate isomerase (GPI) antibodies (Abs) are candidates for arthritogenic Abs because they directly induce arthritis in mice. High titers of anti-GPI Abs are found in some RA patients with severe forms. The aim of this study was to analyze the role of IgG, including anti-GPI Abs, in the joints of RA patients. Synovial tissue was obtained from 6 patients with RA (3 anti-GPI Abs- positive and 3 anti-GPI Abs- negative) and compared histologically and immunohistochemically for IgG and C3 deposition. IgG fractions were separated from the sera of anti-GPI Abs-positive RA patients and healthy subjects, and injected into the metacarpophalangeal joints of 4 cynomolgus monkeys. On day 16, the joints were harvested and examined histologically and immunohistochemically. The expression of the C5a receptor (C5aR) molecule in the synovium was quantified by real-time PCR using cDNA from the monkeys' joints. The synovia of anti-GPI Abs positive RA patients showed diffuse infiltration of cells, including mast cells, and strong deposition of IgG and C3. In monkeys, IgG from RA patients, including anti-GPI Abs, resulted in recruitment of granulocytes and mononuclear cells, strong deposition of IgG on the articular surface, and overexpression of C5aR, but no joint swelling. No infiltrated cells or IgG deposition were observed in monkeys injected with IgGs from healthy subjects. Our results suggest that IgG fraction from RA patients, including that of anti-GPI Abs, may play a role in the synovitis of RA, although the pathogenesis of human anti-GPI Abs is still uncertain. PMID- 16211247 TI - Soluble amyloid beta-peptide and myelin basic protein strongly stimulate, alone and in synergism with combined proinflammatory cytokines, the expression of functional nitric oxide synthase-2 in normal adult human astrocytes. AB - The accumulation of amyloid beta (Abeta)-peptides and their collection in fibrillar plaques in the human brain are believed to be responsible for Alzheimer's disease. The major neuron killers in the Alzheimer brain include proinflammatory cytokines and NO made by NOS-2 (inducible nitric oxide synthase 2). We have determined the effect of a soluble Abeta peptide, Abeta(1-40), on the expression of NOS-2 in astrocytes using a novel model system consisting of pure cultures of cells from adult human brains that, after the first three passages in vitro, become stably locked into the normal astrocytic phenotype like their counterparts in the adult human brain. Abeta(1-40) alone stimulated quiescent astrocytes to start expressing functional NOS-2 and dumping NO into the culture medium during the next 4 days. But adding three of the proinflammatory cytokines commonly produced in the Alzheimer brain--IFN-gamma, IL-1beta, and TNF-alpha- along with Abeta(1-40) more than trebled NOS-2 expression and doubled NO production. In view of the possibility of myelin breakdown in the Alzheimer brain, we also tested the capability of myelin basic protein (MBP) to stimulate NO production using human astrocytes. We found that MBP mimicked the ability of Abeta(1-40) to induce cells to release NO and adding the cytokine triad along with MBP more than doubled NO production and release. Thus, it appears that Abeta peptides and MBP can join forces with proinflammatory cytokines to enhance the NO mediated killing of neurons in the Alzheimer brain. PMID- 16211248 TI - Nuclear localization of a novel protein, RGPR-p117, in cloned normal rat kidney proximal tubular epithelial cells. AB - A novel protein RGPR-p117 was discovered as a regucalcin gene promoter region related protein that binds to the TTGGC motif. The role of RGPR-p117 in cell function is unknown. The nuclear localization of RGPR-p117 was investigated using cloned normal rat kidney proximal tubular epithelial NRK52E cells in vitro. RGPR p117 mRNA was expressed in NRK52E cells, and its expression was stimulated by culture with parathyroid hormone (10(-7) M) or phorbol 12-myristate (10(-6) M). RGPR-p117 was found to localize in the cytoplasm and nucleus with immunocytochemical and Western blot analysis using HA-RGPR-p117/phCMV2 transfected NRK52E cells. Overexpression of HA-RGPR-p117 was found to have a significant stimulatory effect on regucalcin mRNA expression in NRK52E cells. This study demonstrates that RGPR-p117 is localized in the nucleus of kidney cells, and may be involved in gene expression. PMID- 16211249 TI - Detection of HPV 52, 58 and 87 in cervicovaginal intraepithelial lesions of HIV infected women. AB - HIV positive or otherwise immunosuppressed patients are susceptible to cervicovaginal infections with a wide spectrum of HPV types. The aim of our study was to investigate the distribution of HPV in squamous intraepithelial lesions (SIL) with regard to HIV infection. We evaluated the HPV status in 20 HIV positive women with cytologically assessed SIL (11 high grade, 9 low grade) in relation to clinical and histological/cytological findings. Twenty HIV negative patients (15 high grade, 5 low grade SIL) served as a control. HPV typing was performed by polymerase chain reaction followed by PCR-ELISA (HPV 6/11, 16/18, 31/33, 40, 45, 52, 58) or sequence analysis of the amplicon (HPV 73, 87). HPV 52 was the most common type in the HIV positive group (8 HIV positive cases vs. 1 HIV negative case). HPV 16/18 was found in 6 HIV positive and 11 HIV negative patients. Further types detected in HIV positive patients were HPV 40, 58, 73 and 87 (one case each). No correlation was found between the HPV status and the CD4+ count or the grading of SIL. Persisting HPV infection with recurrence of SIL was documented in 5 cases after initial therapy of HPV positive lesions (HPV 87, 73, 58, 31/33, 16/18). HIV infected patients reveal a wider spectrum of HPV types in cervicovaginal SIL than HIV negative women. Especially HPV 87 and its relation with HIV infection and development and persistence of SIL needs further investigation. Our results indicate the inclusion of otherwise rare HPV types in screening programs for HIV positive and immunosuppressed patients. PMID- 16211250 TI - Transfecting the multidrug resistance protein 2 gene improves transcellular organic anion transport. AB - Eisai hyperbilirubinuria rats (EHBRs) lack functionally active multidrug resistance protein 2 (Mrp2), which causes impaired biliary excretion of numerous organic anions. We previously reported that Mrp3 expression is enhanced while organic anion transporting polypeptide 1 (Oatp1) or Oatp2 expression is reduced in the liver of EHBRs. Mrp3 mediates basolateral efflux of organic anions but not canalicular export. In this study, we transfected the human MRP2 gene into the liver of EHBRs and evaluated whether its transfection improves transcellular transport of organic anions in hepatocytes of EHBRs. The protein expression vector (pDEST26) including the full-length human MRP2 cDNA was developed. This vector was mixed with the hemagglutinating virus of Japan (HJV) envelope protein and transfected into the liver of EHBRs via the portal vein. Expression of Mrp3, Oatp1 and Oatp2 was evaluated by reverse transcription-polymerase chain reaction and Western blot analysis. mRNA and protein expression of MRP2 were detected in hepatocytes from transfected EHBRs. The serum-conjugated bilirubin level in EHBRs decreased to a normal level (35.7 to 6.4 micromol/l) with the expression of human MRP2. The change in expression of Mrp3, Oatp1 and Oatp2 in the liver of EHBRs was normalized by transfecting MRP2. Transfection of human MRP2 was performed using the HJV envelope protein. Transfection of MRP2 is useful for improving the transcellular transport of organic anions in the livers of EHBRs. PMID- 16211251 TI - A novel single-nucleotide substitution, Glu 4 Lys, in the leukotriene C4 synthase gene associated with allergic diseases. AB - Cysteinyl leukotrienes (cysLTs) play important roles in bronchial asthma, and can mediate bronchial smooth muscle constriction and increase mucous secretion, vascular permeability and cellular infiltration. We identified a novel heterozygous single-nucleotide substitution 10G>A (Glu 4 Lys) in the first exon of the leukotriene C4 synthase gene (LTC4S). This substitution was detected in 5 of 141 allergic patients, but not in 110 nonallergic subjects. There was a difference in the Glu 4 Lys frequency between the allergic patients and nonallergic subjects (Fisher's exact test, p=0.0460). The five patients with Glu 4 Lys had allergic diseases such as bronchial asthma and/or allergic dermatitis. Furthermore, a familial analysis of Glu 4 Lys revealed a link with allergic diseases. Thus, our results suggest that Glu 4 Lys in the LTC4S might be associated with allergic diseases. PMID- 16211252 TI - Induction of heat shock protein 72 in C6 glioma cells by methyl jasmonate through ROS-dependent heat shock factor 1 activation. AB - Salicylate and jasmonates are two different types of plant hormone that play critical roles in plant defense responses against insect herbivores and microbial pathogens, through activating defense genes. These two natural products have been shown to have similar activities in animal cells: the compounds are able to induce cell cycle arrest or apoptosis in a variety of human cancer cells including those of colon, prostate, breast, and leukemia, suggesting the chemicals may potentially be a novel class of anti-cancer drugs. Since sodium salicylate can induce the heat shock response in animals, we examined the effects of jasmonates on the heat shock response in C6 glioma cells. Here, we show that brief exposure to methyl jasmonate (MeJA), but not to jasmonic acid, induces heat shock protein 72 (HSP72), but not HSP73 and HSP90, via heat shock factor I (HSF1) activation in C6 glioma cells without affecting cell viability. Intracellular H2O2 and O2-, and mitochondrial ROS were prominently increased in response to 5 mM MeJA in C6 cells. MeJA-induced HSP72 expression, HSF1 DNA binding, and human HSP70 promoter-driven CAT activity were prevented by N-acetyl-L-cysteine (a general antioxidant), catalase (a specific antioxidant for H2O2), and sodium formate (an inhibitor of OH.), but not by Rac1 dominant negative mutant Rac1N17 and diphenyleneiodonium (a NADPH oxidase inhibitor), indicating that MeJA induces HSP72 expression though HSF1 that is activated via Rac1-NADPH oxidase-independent ROS production pathway. These results suggest that the plant stress hormones share the ability to induce heat shock response in animal cells. PMID- 16211254 TI - Identification of novel mutations of IRF6 gene in Chinese families with Van der Woude syndrome. AB - Van der Woude syndrome (VWS) is an autosomal dominant disorder of syndromic clefts clinically characterized by lower lip pits, cleft lip and/or palate, hypodontia. Mutations in the IRF6 gene have recently been found to cause VWS and more than 70 mutations have been reported. However, genotype distribution and prevalence of IRF6 mutations underlying Chinese are largely unknown. In the present study, we report on four Chinese families with VWS. Considerably variable clinical phenotypes were observed between and within each family. By direct sequencing, three novel mutations (Y111H, S407fsX436, F165fsX166) as well as a recurrent mutation (R400W) were identified. In contrast to the IRF6 mutations reported in Caucasians, the majority of these mutations occurred at a run of 1- or 2-base repetitive sequence unit, and localized neither in the conserved DNA binding domain nor in the Smad-interferon regulatory factor-binding domain (SMIR). Therefore, our results indicate the existence of other putative IRF6 regions that are predisposed to mutations. Repeated nucleotides in the IRF6 coding regions may increase the instability and chance of DNA replication errors, and are prone to be potential mutation hot-spots. PMID- 16211253 TI - Implication of reactive oxygen species, ERK1/2, and p38MAPK in sodium salicylate induced heat shock protein 72 expression in C6 glioma cells. AB - Sodium salicylate, one of anti-inflammatory agents, is known to partially induce the heat shock response: it stimulates the DNA-binding of heat shock factor 1 (HSF1) without inducing heat shock gene expression. Here we show that when C6 glioma cells are recovered from sodium salicylate treatment, they highly induce heat shock protein 72 (HSP72), but not HSP73 and HSP90, demonstrating that salicylate-induced inert HSF1 can be fully activated into a transcriptionally competent form by sodium salicylate recovery (SR)-specific mechanism. Fluorescent analysis using 2',7'-dichlorodihydrofluorescein diacetate revealed that sodium salicylate enhanced reactive oxygen species (ROS) production. N-acetyl-L-cysteine (NAC, a ROS scavenger) completely suppressed SR-induced HSP72 synthesis and HSP72 promoter-driven CAT reporter gene transcription as well as salicylate-induced HSF1-DNA binding, indicating a critical role(s) of ROS in the SR-induced HSP72 gene regulation. We also show that treatment of C6 cells with sodium salicylate activated p38MAPK and inactivated ERK1/2 in a ROS-independent manner and activities of these protein kinases returned during recovery period to the control level. Inhibiting p38MAPK and ERK1/2 with the p38MAPK inhibitors (SB203580 and SB202190) and the MEK1/2 inhibitor (PD98059 and U0126) or with expression of dominant negative p38MAPK and ERK1/2 abolished SR-induced HSP72 synthesis and HSP70 promoter-driven CAT activity. However, sodium salicylate induced HSF1-DNA binding was not affected by the p38MAPK inhibitor or the MEK1/2 inhibitor. These findings suggest that sodium salicylate partially activates HSF1 via ROS production and p38MAPK activation and the salicylate-induced inert HSF1 can be fully activated into a transcriptionally competent form by the ERK1/2 signaling pathways that are activated independently of ROS during SR. PMID- 16211256 TI - Expression of Y-Box binding protein-1 following hypericin-mediated photodynamic therapy in well-differentiated nasopharyngeal cancer in vivo. AB - The Y-Box binding protein, YB-1, belongs to a group in the DNA and RNA binding protein family. YB-1 is known to be expressed in a number of different cancers and reported to protect cells against DNA-damaging agents such as ultraviolet light. The purpose of this study was to determine if YB-1 levels are altered in photodynamic therapy (PDT)-treated nasopharyngeal cancer (NPC) cells. In this report, we show for the first time that YB-1 is expressed at both the mRNA and protein levels in well-differentiated NPC in vivo. YB-1 mRNA expression in tumor tissues from a murine well-differentiated HK1/NPC model showed no significant difference in mRNA levels following hypericin-PDT. Localization of the YB-1 protein by immuno-histochemistry revealed the presence of cytoplasmic YB-1 in the more mature and better differentiated cells of the untreated group, as well as cells that survived PDT treatment. Expression of the YB-1 protein may possibly influence the cellular stress response to hypericin-PDT and protect the cells from phototoxicity. PMID- 16211255 TI - Quantitative detection of telomerase activity by real-time TRAP assay in the body fluids of cancer patients. AB - Real-time TRAP assay was developed to improve the sensitivity and quantitative detection of telomerase activity in the body fluids of cancer patients. The sensitivity and clinical significance of the real-time TRAP assay was investigated. Real-time PCR protocol was modified by using ACX primer and SYBR green mixture from the process of TS primer extension. Real-time TRAP showed high correlation (r2=0.843, p=0.001) and sensitivity (25 times higher) compared to conventional TRAP. Of 164 samples, there were 8 positives in cytology (4.9%), 7 (4.3%) using the conventional TRAP, and 41 (25%) using real-time TRAP. In cytology positive samples, real-time TRAP showed a higher positivity than conventional TRAP (75% vs 63%) suggesting a higher sensitivity in the body fluids. There was a tendency towards a longer progression-free duration in telomerase negative patients than in positive patients, as determined by conventional and real-time TRAP. The diagnostic interval between the first positivity documentation and clinical progression was short in the order of real time TRAP, conventional TRAP and cytology. In conclusion, real-time TRAP assay can detect telomerase activity at an earlier phase of cancer progression and can replace conventional TRAP assay for detecting the telomerase activity in body fluids. PMID- 16211257 TI - Tumor necrosis factor alpha promotes an osteoblast-like phenotype in human aortic valve myofibroblasts: a potential regulatory mechanism of valvular calcification. AB - Valvular calcification during calcific aortic stenosis is associated with morphological features of bone formation and expression of various bone associated proteins, which are both associated with marked leukocyte infiltration of the calcified valve areas. The pro-inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) is abundantly present in areas of leukocyte infiltration in stenotic aortic valves. We therefore hypothesized that valvular calcification might be actively regulated by an inflammatory process involving TNF-alpha. Upon stimulation with TNF-alpha, human aortic valve myofibroblasts cultured under mineralizing conditions showed an increased formation of calcified, alkaline phosphatase (ALP)-enriched cell nodules, ALP activity, concentration of the bone type ALP isoenzyme, and concentration of osteocalcin, all of which are markers of an osteoblast-like cellular phenotype. By electrophoretic mobility shift assay, DNA binding of the essential osteoblastic transcription factor Cbfa-1 was increased compared to untreated controls. These results support the concept that aortic valve calcification is associated with an osteoblast-like phenotype of local myofibroblasts. In addition, the data demonstrate direct mechanistic evidence that aortic valve calcification may be actively regulated by an inflammatory process involving TNF-alpha. PMID- 16211258 TI - Role of protein kinase C in expression of granulocyte-colony stimulating factor and granulocyte macrophage-colony stimulating factor in lung cancer cells. AB - We examine the role of protein kinase C (PKC) pathways in the constitutive expression of granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF) in lung cancer cells. Two cell lines, OKa-C-1 and MI-4, constitutively produce an abundant dose of G-CSF and GM CSF. The PKC activator phorbol 12-myristate 13-acetate (PMA) stimulated the production of GM-CSF in a dose-dependent manner and reduced G-CSF in the cell lines. The PKC inhibitor staurosporine had effects opposite to those of PMA in the cell lines. Another PKC activator (4beta-phorbol 12, 13-dibutyrate) and six specific PKC inhibitors (bisindolylmaleimide I, calphostin C, chelerythrine chloride, Go 6976, PKC inhibitor 19-27, and Ro-32-0432) also worked as well as PMA and staurosporine, respectively. The induction of GM-CSF expression via PKC activation was mediated by the activation of nuclear factor-kappaB. The induction of G-CSF expression via PKC inhibition was mediated by p44/42 mitogen-activated protein kinase and c-Jun N-terminal kinase pathway signaling. GM-CSF may accelerate cell growth and inhibit cell death via PKC activation in the cell lines. G-CSF also seems to reverse growth suppression and cell death induced by PKC inhibition. PMID- 16211259 TI - Marked telomere fluctuation of leukocytes during graft-versus-host disease in allogeneic stem cell transplantation. AB - Immune dysfunction after allogeneic stem cell transplantation (SCT) is closely associated with cell turnover of lymphocytes and homeostasis of hematopoietic stem cells. Telomeres, repetitive sequences (TTAGGG)n on the end of linear chromosomes, reflect the mitotic history of stem cells. Using telomere fluorescence in situ hybridization (FISH) and flow cytometry (flow-FISH), we measured telomere length in lymphocytes and neutrophils at various intervals to analyze the relationship between telomere length change and clinical features in 5 patients who underwent allogeneic bone marrow transplantation. During the first year after allogeneic stem cell transplantation, a marked fluctuation of telomere length in peripheral blood leukocytes was observed in all recipients, and in 3 patients there was a reduction of telomere length during chronic graft-versus host disease (GvHD) or during post-transplant lymphoproliferative disorder. The reduction of telomere length during GvHD was evident in lymphocytes and neutrophils, but telomere length in neutrophils tended to recover earlier than that observed in lymphocytes. The rapid reduction of telomere length in leukocytes during GvHD was too extensive to be explained by the end-replication problem, suggesting the presence of a telomerically unstable hematopoietic condition after transplant in vivo. PMID- 16211260 TI - Differential-expression and tyrosine-phosphorylation profiles of caveolin isoforms in human T cell leukemia cell lines. AB - Caveolin, the essential structural component of caveolae, serves as a scaffolding protein onto which signaling molecules are assembled, and functions as a negative regulator for signal transduction. Caveolin-1 and -2 are expressed in most cell types, but are not expressed in normal blood cells and cell lines. We previously demonstrated that caveolin-1 is expressed in a panel of human leukemia cell lines that show an activated T cell phenotype. In that study, we detected two caveolin bands by Western blotting using a polyclonal antibody (pAb) reacting with caveolin-1, -2, and -3. We identified caveolin-1alpha by its large molecular weight, but did not discriminatively detect other caveolin families. Since anti caveolin-1 monoclonal antibody (mAb) was reported not to detect caveolin-1 in some cases, here we developed a sensitive method for the discriminative detection of caveolin-1, -2, and -3 by modified Western blotting. Caveolins were solubilized using a two-step procedure and detected by immunoprecipitation with a pAb to caveolins followed by Western blotting with mAbs specific to each caveolin. Using this method we detected caveolin-1beta, -2alpha and -2beta, but not caveolin-3 in the leukemia cell lines. Caveolin-1alpha, which was identified by pAb, was not detected by this method. We show here that caveolin-1alpha and 2alpha, but not caveolin-1beta and -2beta, are tyrosine phosphorylated. This modification is likely to cause the lack of reactivity of caveolin-1alpha to the mAb, and suggests a possible close relationship to cell activation. PMID- 16211261 TI - Synthesis and evaluation of trimethoxyphenyl isoxazolidines as inhibitors of secretory phospholipase A2 with anti-inflammatory activity. AB - A series of trimethoxyphenyl isoxazolidine derivatives, 5a(i-v) and 5b(i-v), bearing different constituents at the 5th position of the isoxazolidine ring were synthesized and evaluated in vitro and in vivo for their inhibitory activity against purified group I and II phospholipase A2 (PLA2) enzymes from snake venom and human inflammatory synovial fluid. Irrespective of modification to the pharmacophore (isoxazolidine ring), they exhibited greater specificity for group II PLA2. The length of alkyl or aryl group at the 5th position, which alters the hydrophobic and aromatic property, was responsible for enhancing the inhibition towards PLA2 enzymes. All of the compounds quench the fluorescent property of the purified PLA2 enzyme, and quenching increases with the increase in length of alkyl or aryl group. The inhibitory effect of compounds appeared to be due to the direct interaction of compounds with the enzyme. Inhibition is substrate dependent, and the inhibitor likely competes with the substrate for the same binding site of the enzyme. The IC50 value for the most potent interacting inhibitor 5b(v) was 54.8 microM. The most active interacting compounds 5a(v) and 5b(v) from in vitro inhibition of PLA2 activity showed similar potency in in vivo neutralization of PLA2-induced mouse paw edema and hemolytic activity. PMID- 16211262 TI - LASS6, an additional member of the longevity assurance gene family. AB - Longevity assurance genes (LAGs) represent a subgroup of the homeobox gene family. Five mammalian homologs have been reported, and the corresponding proteins have previously been investigated with respect to their key role in ceramide synthesis. However, members of the LAG family have been shown to be involved in cell growth regulation and cancer differentiation. In an effort to characterize additional members of the LAG family, we have screened the latest releases of genomic databases and report on the bioinformatic characterization of yet another member, LAG1 longevity assurance homolog 6 (LASS6). Like other LAG family members, the LASS6 protein contained a homeodomain and LAG1 domain. In phylogenetic analyses, it displayed highest homology to LASS5. The corresponding gene was localized to human chromosome 2q24.3, spanning a rather large genomic region of 318 kb. Orthologous sequences in mouse and zebrafish suggested a conservation of LASS6 in vertebrates as the protein and corresponding genomic sequences were highly conserved. LASS6 expression was analyzed in silico, and the gene was shown to be broadly expressed in a wide range of tissues. Furthermore, available microarray data suggested a role in cancer differentiation and early embryonic development. PMID- 16211263 TI - Maintenance of open DNA base pairs through histone acetylated lysine-purine interaction leading to transcriptional activation: a proposed mechanism. AB - Post-translational acetylation of lysines of the histone N-terminal tails is known to induce transcriptional activation, and thus plays a major role in gene regulation. A mechanism for this effect is suggested by our recent finding that the initial 'solvation' network, which is formed around the purines of base pairs immediately following their opening, has the tendency to be preserved. The experiments involved studying the solvation of nucleosides in water-alcohol mixtures; these systems model the hydrophobic/hydrophilic effects that participate in the interaction between histone-tail amino acid residues and nucleosomal DNA base pairs following their opening by the action of DNA-binding proteins in conjunction with remodeling complexes. A highly amphiphilic molecular environment, which has a four-carbon aliphatic chain and hydrogen bond accepting and donating capabilities, was found to promote this phenomenon. Acetylated lysines, unlike lysines, possess these properties and thus constitute such a molecular environment. Opened base pairs are maintained by direct interactions of their purines with acetylated lysines that are selectively recognized by the bromodomains of the recruited remodeling complexes. This maintenance is in accord with the reported large enhancements of i) transcription factor binding to nucleosomal DNA and (ii) the degree of DNA flexibility, which are associated with histone-tail acetylation. This mechanism suggests a way to possibly establish an orderly sequence of transcriptional events despite the reported prevailing stochasticity. It also suggests an approach for the artificial control of transcriptional activation through the use of highly amphiphilic small molecules that would mimic the action of acetylated lysines; in view of the known association of aberrant transcription with cancer, this approach holds promise for advances in cancer etiology and treatment and represents an alternative to the approach that currently uses histone deacetylase inhibitors (some of which are in clinical trials). PMID- 16211264 TI - Phylogeny of the TMEM16 protein family: some members are overexpressed in cancer. AB - TMEM16 proteins are found in all eukaryotes and have eight putative transmembrane domains with NH2 and COOH termini located on the luminal side of the vesicle or plasma membrane. Nine homologues exist in humans and mice. Several of the human genes are overexpressed in cancer and could be valuable tumor markers, especially in profiling gene expression with microarrays. In Drosophila, homologues are involved in chromosome separation. In Baker's yeast, the one homologue is expressed exclusively in the bud and involved in osmotic regulation. In sea urchin embryos, the protein is associated with nuclei. In mammals, the functions of TMEM16 proteins are still unknown. We predict that the TMEM16 proteins will gain importance in the study of normal and malignant tissues. This report presents the first comprehensive phylogeny of TMEM16 proteins. The two phylograms show that these proteins fall into distinct families, which are differentiated from each other in all animal lineages. Here, all previous studies on these proteins are compiled in table form with the hope this compilation will facilitate future work on these genes and their protein products. PMID- 16211265 TI - Inhibition of proteasome activity in Gleditsia sinensis fruit extract-mediated apoptosis on human carcinoma cells. AB - The anomalous fruit extract of Gleditsia sinensis (GSE) was shown to possess anticancer potential on various solid tumour and leukaemia cell lines in vitro. We have recently demonstrated that the mitochondrial-dependent apoptotic pathway including mitochondrial membrane potential depolarization, changes in the level of reactive oxygen species and activation of caspase 3 were recruited in GSE induced apoptosis. Whether receptor-dependent APO-1/Fas apoptotic pathway is also involved remains uncertain. Using two solid tumour cell lines, the HepG2 hepatoblastoma carcinoma cells and MDA-MB231 breast cancer cells, we demonstrated that the Fas ligand and Fas receptor protein levels did not have significant variation after GSE incubation. Caspase 8 activity increased only weakly when compared with that of caspase 3. The chrymotrypsin-like activity of proteasome was partially inhibited up to 30-40% when compared with the untreated control. Taken together, we believe that GSE- mediated apoptosis on HepG2 and MDA-MB231 carcinoma cells is mainly dictated by the mitochondrial-dependent pathway while inhibition of proteasome activity may also be involved in GSE-induced apoptosis. PMID- 16211266 TI - Antiproliferation and induction of cell death of Phaffia rhodozyma (Xanthophyllomyces dendrorhous) extract fermented by brewer malt waste on breast cancer cells. AB - Astaxanthin has been shown to have antiproliferative activity on breast cancer and skin cancer cells. However, the high cost of production, isolation and purification of purified astaxanthin from natural sources or chemically synthetic methods limit its usage on cancer therapy. We show that astaxanthin could be produced by fermentating the Phaffia rhodozyma (Xanthophyllomyces dendrorhous) yeast cells with brewer malt waste using a 20 L B. Braun fermentor. The percentage composition of astaxanthin from the P. rhodozyma was >70% of total pigment as estimated by the high performance liquid chromatographic analysis. Furthermore, the antiproliferative activity of this P. rhodozyma cell extract (PRE) was demonstrated on breast cancer cell lines including the MCF-7 (estrogen receptor positive) and MDA-MB231 (estrogen receptor negative) by using the [3 (4,5-dimethylthiazol-2-yl)-5-(3-arboxymethoxyphenyl)-2- (4-sulfophenyl)-2H tetrazolium] (MTS) assay. No apoptotic cell death, but growth inhibitory effect was induced after 48 h of PRE incubation as suggested by morphological investigation. Anchorage-dependent clonogenicity assay showed that PRE could reduce the colony formation potential of both breast cancer cell lines. Cell death was observed from both breast cancer cell lines after incubation with PRE for 6 days. Taken together, our results showed that by using an economic method of brewer malt waste fermentation, we obtained P. rhodozyma with a high yield of astaxanthin and the corresponding PRE could have short-term growth inhibition and long-term cell death activity on breast cancer cells. PMID- 16211267 TI - Regulation of tyrosine phosphorylation during the CD40-mediated rescue of Ramos BL B cells from BCR-triggered apoptosis. AB - The regulation of tyrosine phosphorylation is essential for BCR-triggered cellular responses during the selection process in the germinal centres. We were interested in examining the temporal regulation of tyrosine phosphorylation following CD40 cross-linking of anti-IgM-triggered Ramos-BL B cells. CD40 co stimulation of anti-IgM-treated Ramos-BL B cells rescued them from growth inhibition and apoptosis, even when anti-CD40 Abs were added up to 12 h after the cross-linking of the BCR. The initial up-regulation of tyrosine phosphorylation triggered by BCR cross-linking is followed by tyrosine dephosphorylation after 12 h of stimulation, coinciding with pro-caspase-3 processing and PARP cleavage. We find that CD40 co-stimulation rescues BCR-triggered Ramos-BL B cells only before the irreversible inhibition of tyrosine kinase activity after 12 h of BCR cross linking and that this is coupled with up-regulation of tyrosine phosphorylation; thus demonstrating the importance of the late regulation of tyrosine phosphorylation for CD40-mediated rescue of Ramos-BL B cells from BCR-triggered G1 growth arrest and apoptosis. PMID- 16211268 TI - UV-induced NF-kappaB activation and expression of IL-6 is attenuated by (-) epigallocatechin-3-gallate in cultured human keratinocytes in vitro. AB - Ultraviolet (UV) radiation from the sun is widely considered as a major cause of human skin photoaging and skin cancer. UV radiation-induced proinflammatory cytokines mediated by NF-kappaB reportedly play important roles in photoaging and cancer. NF-kappaB and cytokines have been thus perceived as molecular targets for pharmacological intervention. With an increasing amount of knowledge of the actions of green tea extracts at cellular and molecular levels, the beneficial effect of drinking green tea has become well recognized if not completely accepted. The components in green tea have even been added to skin-care products unregulated, while the molecular mechanisms of the actions of those components on human skin are being unraveled. Using cultured human keratinocytes, we investigated the effects of (-)-epigallocatechin-3-gallate (EGCG), a major polyphenolic constituent in green tea, on UV-induced activation of transcription factor NF-kappaB and proinflammatory pathway by measuring nuclear translocation of NF-kappaB and IL-6 secretion in vitro. Immunohistochemical and Western blot analysis and ELISA indicated that both nuclear p65 and secreted IL-6 were significantly (p<0.05) induced by UVB (20, 30 mJ/cm2) and UVA irradiation (10, 20 J/cm2). NF-kappaB nuclear translocation and IL-6 secretion induced by UVB and UVA were dramatically inhibited by treatment of EGCG. FACS analysis showed that EGCG also inhibited UVB-induced apoptosis. EGCG recovered UV-induced loss of anti apoptotic component, bcl-2, and inhibited UV-induced apoptotic component, Fas ligand, expression. Collectively, we conclude that EGCG inhibits UVB- and UVA induced proinflammatory pathway and inhibits apoptosis in cultured human keratinocytes in vitro. Our data suggest that EGCG be added to cosmetic or skin care products for prevention from UV-induced skin photoaging if this activity can be further confirmed and no cytotoxicity is reported in human skin in vivo. PMID- 16211269 TI - The order of PNA-FISH-detected chromosomal telomere lengths in human T-cells is rather stable, even under the influence of strong mutagens. AB - The nucleo-protein structure of an intact telomere protects each chromosome from being recognized as a break and subsequently being degraded. The DNA component of this structure, the telomeric repeats, undergo attrition with every cell division, as well as in response to endogenous events, like oxidative stress. Exposure to exogenous damage promotes this process and leads to growth arrest, apoptosis and eventually to malignant transformation. It was thought that the shortest chromosome ends in humans are the most susceptible ones to become dysfunctional telomeres, and have therefore an important role in cell death and cancer. Here, we show that a stable hierarchy exists in the form of a telomere length profile of the whole human karyotype. This rank order is conserved between different human cell types and individuals, is maintained throughout a lifetime, and seems to be genetically determined. As a particular feature, this telomere length profile differs only marginally when normal human cell cultures and malignant transformed cells are compared. The profile is moreover stable when these different human cells are exposed to mutagens such as bleomycin or mitomycin C. From these findings, the question arises if also the stably long telomeres have a basic biological function. PMID- 16211270 TI - Comparative genomics on FGF16 orthologs. AB - We have previously reported comparative genomics analyses on FGF3, FGF4, FGF6, FGF7, FGF8, FGF10, FGF11, FGF17, FGF18, FGF19, FGF20, FGF22 and FGF23 genes. Here, we performed comparative genomics analyses on FGF1, FGF2, FGF5, FGF9, FGF12, FGF13, FGF14, FGF16 and FGF21 genes, and further characterized the FGF16 gene. Chimpanzee FGF16, chicken fgf16, and zebrafish fgf16 genes were identified within NW_121938.1, NW_060344.1, and CR855117.3 genome sequences, respectively. Chimpanzee FGF16 (207 aa), chicken fgf16 (207 aa), and zebrafish fgf16 (203 aa) showed 100%, 89.9%, and 79.2% total amino-acid identity with human FGF16. Because FGF16, FGF9, and FGF20 constitute FGF subfamily without N-terminal signal peptide, we next searched for uncharacterized FGF9 or FGF20 orthologs. Zebrafish fgf9 gene was identified within BX927112.11 genome sequence, and chicken fgf20 gene within NW_060349.1 genome sequence. Although N-terminal part was divergent, middle and C-terminal parts were well conserved among vertebrate FGF16, FGF9 and FGF20 orthologs. Phylogenetic analyses revealed that zebrafish fgf9 and fgf20 were more related to each other than to their chicken or mammalian orthologs. TCF/LEF binding site and TATA box were well conserved among the human FGF16, rat Fgf16, and mouse Fgf16 promoters. Because nuclear complex consisting of TCF/LEF (TCF1, TCF3, TCF4 or LEF1), beta-catenin, PYGO (PYGO1 or PYGO2) and Legless (BCL9 or BCL9L) binds to the TCF/LEF-binding site to up-regulate WNT/beta-catenin target genes, FGF16 gene was characterized as the evolutionarily conserved target of the WNT/beta-catenin signaling pathway, just like FGF18 and FGF20 genes. These facts indicate that FGF16, FGF18 and FGF20 are pharmacogenomics targets in the field of oncology and regenerative medicine. PMID- 16211271 TI - Dual FISH analysis of benign and malignant tumors of the salivary glands and paranasal sinuses. AB - To date, the underlying genomic changes in benign and malignant tumors of salivary-gland and paranasal-sinus origin are poorly understood. This is due in part to the low incidence of these tumors and the enormous histological variety of tumors within this head and neck region. We examined 58 of these tumors (14 adenoid cystic carcinomas, 9 adenocarcinomas, 5 cylindrical carcinomas, 11 pleomorphic adenomas, and 19 inverted papillomas) by dual fluorescence in situ hybridization (FISH) with centromere-specific probes on six chromosomes (3, 7, 9, 11, 17, and 18) for numerical changes. In adenoid cystic carcinomas, monosomy of chromosome 17 and polysomy of chromosomes 3, 9 and 11 were most frequently encountered. In adenocarcinomas, monosomy of chromosome 17 and polysomy of chromosomes 7 and 11 were most frequent. In cylindrical cell carcinomas, polysomy of chromosomes 7, 9, 11 and 17 was present in the majority of tumors. Disomy is rare, even in benign tumors. Polysomy is more frequent in malignant tumors than in benign. Tetrasomy is found almost only in malignant tumors. In summary, the occurrence of polysomy might reflect a step towards malignancy in tumors of the salivary glands and paranasal mucosa. Polysomy of chromosome 11 could be defined as typical for all investigated histological types of malignant tumor in this region of the head and neck. PMID- 16211272 TI - Co-expressed type of ER and HER2 protein as a predictive factor in determining resistance to antiestrogen therapy in patients with ER-positive and HER2-positive breast cancer. AB - The purpose of this study was to analyze whether inter-site variation types on estrogen receptor (ER) and HER2 expression may be a predictive factor for evaluating the effectiveness of endocrine therapy in patients with ER-positive and HER2-positive breast cancer. A total of 366 consecutive women with invasive breast cancer who had undergone curative surgical treatment between 1996 and 2001 were included in this study. ER status was evaluated using the Allred score and HER-2 status was evaluated according to the HercepTest. In ER-positive and HER2 positive tumors, the expression of ER and HER2 was described as the co-expressed type or the differently expressed type using double staining with ER and HER2. Of the 366 patients, 249 (68.1%) were positive for ER and 74 (20.2%) were positive for HER2. ER-positive and HER2-negative tumors were found in 221 patients (60.4%), ER-negative and HER2-negative in 71 (19.4%), ER-negative and HER-2 positive in 46 (12.6%), and ER-positive and HER2-positive in 28 (7.7%). HER2 status was inversely correlated (p<0.01) with ER status. In ER-positive tumors, an inverse correlation between ER and HER2 was also observed. The co-expressed type was found in 10 patients, and the differently expressed type was found in 18. There was no difference in tumor size and nodal involvement between the two types. There was no significant difference in disease-free survival between patients with the co-expressed type tumor and the differently expressed type tumor. In patients with the differently expressed type tumor, those who received antiestrogen therapy showed a significantly better disease-free survival rate than those who did not receive antiestrogen therapy. As for patients with the co expressed type of tumor, no significant difference in disease-free survival was observed between patients with and without antiestrogen treatment. The present study suggests that the co-expressed type of tumour might be a resistant factor to antiestrogen therapy in ER-positive and HER2-positive breast cancer. PMID- 16211273 TI - Effect of bilateral oophorectomy on mammary tumor formation in BRCA1 mutant mice. AB - Germline mutations of breast cancer-associated gene 1 (BRCA1) predispose women to breast and ovarian cancer. It was recently shown that bilateral oophorectomy decreases breast cancer incidence in BRCA1 mutation carriers. To model human BRCA1 carriers, our laboratory has previously created mice with a conditional knockout of the full-length BRCA1 gene in the mammary epithelium combined with a heterozygous knockout of the p53 tumor suppressor gene. These mice developed ER negative mammary tumors and were employed to determine the effects of oophorectomy on tumor formation. Individual knockout mice (BRCA1(Co/Co) MMTV Crep53+/-), following two complete pregnancies, were either oophorectomized or sham treated. Mice were subsequently examined for the development of palpable mammary tumors until they were 12 months of age. Until 135 days post-oophorectomy (255 days of age), the tumor incidence was similar in both oophorectomized and intact mice, approximately 30%. After this time, the increase in tumor incidence was much lower in the oophorectomized mice, while tumor incidence increased in non-oophorectomized mice. The effects of oophorectomy on mammary development in both control and knockout mice were also examined. Oophorectomized mice with a conditional knockout of full-length BRCA1 in conjunction with a loss of one p53 allele exhibited glandular regression with a reduction in the number of mammary epithelial cells following oophorectomy. This study employed a model that may be relevant for testing agents useful against breast cancer in BRCA1 carriers and a subset of sporadic cancers. The data also show that oophorectomy, if performed significantly prior to the time that tumors arise, appears to be quite effective. PMID- 16211274 TI - Cell proliferation of bulky cervical squamous cell carcinoma is strongly associated with a paracrine tissue-specific growth factor(s). AB - To examine the possibility of a cervical squamous cell carcinoma-specific growth factor(s), previously suggested by clinical findings and in vitro culture experiments, we examined in vitro cultures and nude mouse transplantations of cervical squamous cell carcinoma cells obtained from a patient with a bulky cervical tumor without detectable distant metastases. The cancer cells did not proliferate without additional growth factors but remained viable in vitro. Fourteen weeks after transplantation of the tumor cells on their backs, 1 of the 3 nude mice developed large metastatic pelvic tumors without macroscopic metastatic lesions in their lungs or liver. When these pelvic tumor fragments were trans-planted onto the backs of other nude mice, ulcerated back tumors and larger metastatic pelvic tumors, but no macroscopic metastatic lesions in the lungs or liver, were observed. Histopathological examination of these pelvic tumors showed that all were the same squamous cell carcinoma as the primary tumor. These results indicate that cell proliferation of bulky cervical squamous cell carcinoma is strongly associated with a squamous cell carcinoma-specific growth factor(s) in a paracrine manner. PMID- 16211275 TI - A clinical and immunohistochemical study on gastrointestinal mesenchymal tumor (GIMT): RCAS1 as a predictor for recurrence of GIMT. AB - Gastrointestinal mesenchymal tumors (GIMTs) are the most common mesenchymal tumors of the gastrointestinal tract. RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is a cancer cell-surface antigen and has been identified as a prognostic factor in several cancer types. It is thought that tumor cells escape immune attack by expressing RCAS1, which induces apoptosis in receptor positive immune cells. The current study was designed to elucidate the histogenesis of these tumors by using various immunohistochemical markers, and identify parameters that will help to establish the criteria of malignancy in the GIMT. We also discuss the clinicopathological significance of RCAS1 expression in the diagnosis and prognosis of GIMTs. A total of 70 cases of GIMTs were reviewed. Immunohistochemistry was performed between 1990 and 2000, with the avidin-biotin peroxidase complex method on 3 microm-thick sections of formalin-fixed paraffin embedded specimens of GIMTs. Antibodies to the following antigens were used: KIT (CD117), CD34 alpha-SMA, Desmin, cytokeratin, S-100 protein, p53, and RCAS1. Recurrence-free survival analysis was done with Stat View-J 5.0 statistical packages. Univariate analysis for a recurrence-free prognosis demonstrated that antibody detection of p53 expression (p=0.0333) and expression of RCAS1 (p=0.0008) is correlated with a significantly higher potential of recurrence. On multivariate analysis, tumor size and RCAS1 expression were independently and inversely correlated with recurrence-free survival. The expression of RCAS1 has not previously been reported in GIMT; indeed, our study suggests that the expression of RCAS1 is correlated with recurrence not only in carcinomas, but also in mesenchymal tumors. PMID- 16211276 TI - Microsatellite instability and proliferating activity in sinonasal carcinoma: molecular genetic and immunohistochemical comparison with oral squamous cell carcinoma. AB - Sinonasal carcinomas arise from the respiratory epithelium that lines the nasal and paranasal cavities, and are histologically composed of either squamous or cylindrical cell carcinoma. However, molecular analysis with the purpose of distinguishing sinonasal carcinomas from other head and neck squamous cell carcinomas (HNSCCs), which arise from squamous epithelium, has been limited. Moreover, a wide range of frequency of microsatellite instability (MSI) in HNSCC has been reported. Using high-resolution fluorescent microsatellite analysis (HFRMA), we studied microsatellite alterations in 34 patients with sinonasal carcinoma. As a control, 24 oral squamous cell carcinomas were used. MSI was detected in 14 patients with sinonasal carcinoma (41%), but not in any with oral squamous cell carcinoma (p=0.002). Furthermore, in sinonasal carcinoma, 11 out of 17 (65%) T1-T3 sinonasal carcinomas demonstrated MSI, whereas only 3 out of 15 (20%) T4 tumors demonstrated MSI. Immunohistochemically, sinonasal carcinoma showed a higher MIB-1-labeling index and more frequently showed cytokeratin 18 expression when compared with oral squamous cell carcinoma. These findings suggest that sinonasal carcinoma and HNSCC have quite different molecular backgrounds regarding carcinogenesis, and the role of MSI is relatively minor in cases of advanced sinonasal carcinoma. PMID- 16211277 TI - Induction of apoptosis in human salivary gland tumor cells by anti-NCAM antibody. AB - Neural cell adhesion molecule (NCAM) is a type of cell surface glycoprotein and a member of the immunoglobulin superfamily. It has been reported that NCAM may be associated with perineural invasion by malignant salivary gland tumors such as adenoid cystic carcinoma. We have previously demonstrated that NCAM is constitutively expressed in the human salivary gland tumor cell line HSG, in vitro. In the present study, we have aimed to clarify the hypothesis that NCAM mediated inhibition of salivary gland tumor proliferation is caused by homophilic binding and involves the prevention of signal transduction for perineural invasion using HSG cells. NCAM mRNA and protein expression was found to decrease in a dose-dependent manner upon treatment with the anti-NCAM antibody (MAb NCAM) for 24 h. The MTT assay showed a significant reduction in the number of viable HSG cells. Confocal laser microscopy showed that HSG cells underwent apoptosis after treatment with MAb NCAM. The activation of caspases 3, 7 and 9 was observed in HSG cells after treatment with MAb NCAM, thus confirming that apoptosis was induced by the activated caspases. Apaf-1 activity was also detected in HSG cells in a dose-dependent manner after treatment with MAb NCAM. The up-regulation of TGF-beta1-mediated NCAM expression appeared to lead to the activation of homophilic NCAM binding, further accelerating HSG cell proliferation. In addition, the localization of NCAM in adenoid cystic carcinomas (ACCs) was examined using an immunohistochemical method. NCAM was slightly to moderately positive in 9 of 13 cases (69.2%) of ACC. These findings suggest that NCAM is associated not only with a cell-to-cell adhesion mechanism, but also with tumorigenesis, including growth, development and perineural invasion in human salivary gland tumors. PMID- 16211278 TI - No evidence of correlation between the single nucleotide polymorphism of DNMT3B promoter and gastric cancer risk in a Japanese population. AB - DNA methylation is a major epigenetic modification in humans, and aberrant DNA methylation may play an important role in the development of various cancers through the silencing of some tumor suppressor genes. DNMT3B is required for the establishment and maintenance of genomic methylation patterns. The -149 C/T single nucleotide polymorphism (SNP) in the promoter of DNMT3B has been identified. This SNP influences DNMT3B promoter function, with the T allele having greater activity than the C allele, and is associated with an increased risk of lung cancer. The purpose of our study was to investigate the correlation between the DNMT3B promoter polymorphism and the development and progression of gastric cancer. We analyzed the SNP of the DNMT3B promoter in 152 gastric cancer patients and 247 controls from a Japanese population using PCR-RFLP and sequencing analysis, and also studied the association between the genotypes of DNMT3B and clinicopathological parameters among cases. Allelic difference was not found between gastric cancer patients and control subjects at the target site, 149 bp from the transcriptional start site in the DNMT3B gene promoter. Only the T/T genotype was detected in all gastric cancer patients and control subjects. We concluded that there was no association between SNP of the DNMT3B promoter and gastric cancer risk in a Japanese population. PMID- 16211279 TI - E1B-deleted adenovirus replicates in p53-deficient lung cancer cells due to the absence of apoptosis. AB - E1B55K adenovirus codes the protein that inactivates the p53 protein of host cells and facilitates its own cell proliferation. We prepared E1B55K-deleted adenovirus and investigated its cytopathic effects in primary lung cancer cells to evaluate the possibility of its application in gene therapy. A p53 wild-type small cell lung cancer cell line (SBC3) and three p53 mutant cell lines (SBC5, small cell lung cancer; PC3, adenocarcinoma; and EBC1, squamous cell carcinoma) were infected with AxE1AdB, in which the E1B55K gene was deleted, to assess the cytotoxicity and induction of apoptosis. Moreover, the mRNA expression in virus infected cells was studied with cDNA array analysis to investigate the response of cancer cells to the virus infection. As a result, AxE1AdB had no detectable cytopathic effect on SBC3 cells, and SBC3 cells continued to replicate. In contrast, AxE1AdB killed SBC5, PC3, and EBC1 cells with great efficiency. In SBC3, apoptosis of the host cells was observed in the early infection stage; in the p53-mutant cell lines, apoptosis of the cells was restricted. An analysis of gene expression in the host cells indicated a significant increase of Bax mRNA in SBC3 and its deficiency in SBC5. Overall, our results suggest that AxE1AdB replicated selectively in the p53-mutant lung cancer cells and showed strong cytotoxicity against them. In the p53-intact cells, p53 induced apoptosis through Bax expression and prevented virus replication and escalation of the infection. PMID- 16211280 TI - Recurrence and 5-FU sensitivity of stage III/Dukes' C colorectal cancer with occult neoplastic cells in lymph node sinuses. AB - In 72 patients without occult neoplastic cells (ONCs) in their lymph node sinuses, the 5-year relapse-free survival (RFS) rate and overall survival (OS) rate were 71.3% and 69.2%, respectively. In 33 patients with ONCs, the 5-year RFS rate and OS rate were 33.9% and 31.3%, respectively. There was a marked difference of survival between the two groups (p=0.0001 and p=0.0003). The metastatic lymph nodes of the 33 ONC-positive patients had high and low levels of thymidilate synthase (TS) expression in 38.1% (8/21) and 61.9% (13/21) of the recurrence group (n=21), respectively, while high and low levels of dihydropyrimidine dehydrogenase (DPD) expression were found in 38.1% (8/21) and 61.9% (13/21), respectively. In the non-recurrence group (n=12), high and low levels of TS or DPD expression were detected in 58.3% (7/12) and 41.7% (5/12) versus 16.7% (2/12) and 83.3% (10/12), respectively. Patients with high TS and low DPD expression accounted for 9.5% (2/21) of the recurrence group and 50.0% (6/12) of the non-recurrence group (p<0.01). These results suggest that ONCs are clearly associated with the 5-year RFS and OS rates. Unlike the non-recurrence group, the recurrence group of ONC-positive patients with Dukes' C colorectal cancer is unlikely to respond well to treatment with 5-FU plus LV and require combination chemotherapy based on CPT-11 and/or L-OHP. PMID- 16211281 TI - Recurrence and 5-FU sensitivity of stage II/Dukes' B colorectal cancer with occult neoplastic cells in lymph node sinuses. AB - In 103 patients without occult neoplastic cells (ONCs) in the lymph node sinuses, the 5-year relapse-free survival (RFS) rate and overall survival (OS) rate were 90.2% and 91.8%, respectively. In 21 patients with ONCs, the 5-year RFS and OS rates were 34.9% and 62.3%, respectively. There were marked differences of survival between the two groups (p=0.0000 and p=0.0003). In the primary tumors of the 21 ONC-positive patients, high and low TS levels were found in 46.2% (6/13) and 53.8% (7/13) of the recurrence group (n=13), respectively. High and low DPD levels were found in 23.1% (3/13) and 76.9% (10/13), respectively. In the non recurrence group (n=8), high and low TS or DPD levels were found in 75.0% (6/8) and 25.0% (2/8) versus 12.5% (1/8) and 87.5% (7/8), respectively. The percentage of patients with high TS and low DPD levels was 23.1% (3/13) in the recurrence group and 62.5% (5/8) in the non-recurrence group (p=0.07). These results suggest that the presence of ONCs had a clear association with the 5-year RFS and OS rates. The recurrence group of ONC-positive patients with stage II/Dukes' B colorectal cancer was unlikely to be highly responsive to 5-FU-based treatment, thus requiring multi-combination chemotherapy using CPT-11 and/or L-OHP. PMID- 16211282 TI - Chemoradiotherapy for T3 and T4 squamous cell carcinoma of the esophagus using low-dose FP and radiation: a preliminary report. AB - We conducted this study to evaluate the clinical significance of preoperative concurrent chemoradiotherapy (CRT) followed by esophagectomy in the management of T3 and T4 esophageal cancer. Thirty patients with squamous cell carcinoma of the esophagus received CRT followed by surgery. Preoperative CRT consisted of 5 fluorouracil (500 mg/m(2) by 24 h infusion for 5 days), cisplatin (15 mg/m(2) on days 1-5), and concurrent radiotherapy (a total dose of 40 Gy delivered in daily fractions of 2 Gy, 5 times per week). Esophagectomy was planned for 4-6 weeks after treatment and restaging. All 30 patients completed preoperative CRT. A clinical response (PR+CR) of the primary tumor was obtained in 82.8%, and a response of metastatic nodes was seen in 23.1%. Radical resection was possible in 17 of 29 operated patients (58.6%). The postoperative mortality rate was 6.9%, and the hospital mortality rate was 10.3%. Ten out of 29 operated patients (34.5%) had no residual cancer in the resected esophagus, corresponding to pathological CR. The 1-year survival rate was 80.6%, the 2-year survival rate was 62.7%, and the 3-year survival rate was 53.8%. The clinical response group and the R0 or R1 group showed better survival than other patients. Preoperative CRT should be given to patients with squamous cell carcinoma, while esophagectomy remains the standard therapy for responders and has a tolerable mortality. PMID- 16211283 TI - Comparative study of renal cell carcinoma by CGH, multicolor-FISH and conventional cytogenic banding analysis. AB - Using different cytogenetic techniques in combination is crucial to studying the high complexity of genetic rearrangements in tumor cells. The 8 clear cell (cc) and 5 papillary (p) renal cell carcinomas (RCC) were analyzed using multicolor fluorescence in situ hybridization (multicolor-FISH), conventional Giemsa banding (G-banding) and comparative genomic hybridization (CGH) analysis. CGH analysis was carried out with DNA from frozen tissue sections and short-term cultures of primary tumors. Using CGH analysis, both tissue sections and cell cultures of ccRCC showed the typical chromosomal changes such as the loss of 3p, 4q, 6q, 8p, 9q, 14 and a gain of 5q and 7. Most imbalances detected by CGH in cell culture could be deciphered by multicolor-FISH and G-banding analysis as unbalanced trans locations t(3;6)(p11.1;p11.1), t(8;14)(p11.1;q11.1), t(3;5) (p14;q21-22), t(1;15)(p11;q11.1), t(3;15)(p11;q11.1)t(8;17) (p11.1;q11.1), t(8;17)(q22;p11.1). Only one balanced trans-location t(9;18)(q34;q11.1) was shown in ccRCC. CGH of papillary RCC displayed mostly gains of whole chromosomes 7, 12, 16 and 17 and a loss of chromosome Y. There was 1 papillary RCC that displayed a partial gain of chromosome 7, showing an unbalanced translocation t(7;11)(q11.1;q25). The balanced translocations t(2;9)(q11.1;q34) and t(7;15) (q22 approximately 31;q21 22) were registered in pRCC. The combined analysis of RCC by different methods allowed a more accurate characterization of the complex karyotypes of tumor tissue, and offered a comprehensive description of given tumors. PMID- 16211284 TI - Gene expression profiling of the bone marrow mononuclear cells from patients with myelodysplastic syndrome. AB - The gene expression pattern of bone marrow mononuclear cells (BMNCs) from 10 patients with myelodysplastic syndrome (MDS) was studied by two-color cDNA microarray techniques. To confirm the microarray results, a semiquantitative RT PCR was performed to analyze gene expression in fifty additional MDS patients. Ninety-five genes were shown to be abnormally expressed in at least five MDS patients compared to normal controls, involving cell growth and differentiation regulation, cell cycle control, signaling and redox; such as thrombospondin 1, phosphatase and tensin homolog, MAD, DNA-damage-inducible transcript 3 (DDIT3), ets variant gene 1 (ETV1), and G1 to S phase transition 1. CD36 was also revealed up-regulated in 4 cases. MDS patients in early and advanced stages could be clustered into two distinct groups by hierarchical clustering, wherein a case with isolated thrombocytopenia and other RA patients were clustered into two subgroups. Consistent expression patterns of 3/5 (60%) genes were confirmed by semiquantitative RT-PCR. Further analysis showed the different transcript levels of RNAHP, DDIT3 in patients with MDS in different stages, AML, and normal controls. Meanwhile, the different significance of RNAHP and ETV1 expression was revealed between RA and untypical anaplastic anemia, iron deficiency anemia, and megaloblastic anemia patients. We propose that the technology of microarray may reveal the intrinsic molecular features and the expression levels of RNAHP, DDIT3, and ETV1 may provide useful markers for the diagnosis of MDS. PMID- 16211285 TI - Tenascin C and epidermal growth factor receptor as markers of circulating tumoral cells in bladder and colon cancer. AB - Tenascin C has been recently suggested as a tumor marker, however, its levels in serum has been evaluated only in patients with head and neck cancer and melanoma. In this study, we investigated Tenascin C expression in blood samples from colorectal and bladder cancer patients, compared to that of epidermal growth factor receptor (EGFR), a known circulating tumor marker in these cancer types. RT-PCR specific for Tenascin C and EGFR was performed on RNAs extracted from blood samples of 60 patients affected by colon or bladder cancer. We then investigated the statistical association between Tenascin C, EGFR expression and disease-free survival using the Kaplan-Meier method. Furthermore, in order to select which variable between EGFR and Tenascin C was the most predictive for recurrence, a Cox model for proportional risk was applied. Among all patients analysed, a significantly higher disease-free time was found in the group negative for both EGFR and Tenascin C expression; EGFR expression was significantly correlated to disease progression in stages III and IV, whereas in all patients with stage I and II disease Tenascin C correlated better with prognosis. Negative expression of both EGFR and Tenascin C identifies a group of patients with poor tendency to disease recurrence and longer relapse-free time. While Tenascin C expression seems to influence prognosis in patients with low stage disease, EGFR appears a marker of worse prognosis in patients with high staged tumors. PMID- 16211286 TI - Development of an HT29 liver metastases model in nude rats. AB - The liver is the most frequent and fatal site of distant spreading of colorectal cancer. Most liver metastases animal models involve nude mice and an injection of tumour cells through the spleen or portal vein, or orthotopic implantation of tumour cells in the colon wall. The aim of this study was to develop a reliable rat model of liver metastases with human colorectal HT29 cells. Seventy male athymic Rowett nude rats weighing 300+/-30 g were separated into three groups. The first group (n=20) consisted of untreated rats, rats in the second group (n=20) were immunosuppressed by cyclosporin A, and those in the third group (n=30) were irradiated the day before cell grafting. Tumour cells (2 x 10(7)) were subcapsulary injected into the liver, and rats were sacrificed after 60 days. The livers were excised, and tumours were serially sectioned to determine size and volume, then fixed for histological studies. The take-rate was 55% in the first group, 35% in the second and 74% in the third group. The mean volume of tumours in the first group was 537+/-162 mm(3), 613+/-232 mm(3) in the second group and 2949+/-629 mm(3) in the third group. In conclusion, subcapsular injection of the human colonic HT29 cancer cells into the liver of preoperatively irradiated nude rats is a reliable, reproducible and easily obtained model, which should be useful for preclinical studies. PMID- 16211287 TI - 18FDG-positron emission tomography in post treatment evaluation of residual mass in Hodgkin's lymphoma: long-term results. AB - In patients with Hodgkin's lymphoma (HL) at the end of first line therapy an accurate imaging technique with high prognostic value is needed to assess response to treatment and predict those patients who will suffer disease relapse. This technique and its results permit the quick initiation of a second line therapy in patients suffering from a progressive disease or those unresponsive to treatment avoid over-treatment of patients in complete remission or those having a non-active residual disease. We included a (18)FDG-positron emission tomography (PET) scan to the diagnostic set-up to investigate 28 patients following the end of their treatment. Fifteen patients out of the 28 (54%) had positive CT scans while 13 (46%) had negative ones. Eleven patients out of the 15 CT positive (73%) had negative PET scans and no relapse. The remaining four patients (27%) had positive PET scans with only one relapse (25%). With respect to the 13 patients who had negative CT scans, 9 patients (69%) had negative PET scans and no relapse. The remaining 4 patients (31%) had positive PET scans with 3 relapse cases (75%). In our final assessment after a median follow-up period of 45 months, starting from PET execution to the last follow-up, overall sensitivity of the CT and the PET were 25 and 100% respectively, specificity 42 and 83% respectively, positive predictive value (PPV) 7 and 50% respectively, negative predictive value (NPV) 77 and 100% respectively, and accuracy 39 and 86% respectively. In our experience, FDG-PET performed in patients after induction therapy appears to offer important additional information: FDG-PET results are predictors of prognosis giving 100% DFS in PET negative patients and 54% DSF in PET positive patients. PMID- 16211288 TI - Enhancement of susceptibility to Fas-mediated apoptosis in HL-60 cells through down-regulation of cellular FLICE-inhibitory protein by phosphatidylinositol 3 kinase inhibitors. AB - Promyelocytic leukemia HL-60 cells are resistant to Fas-mediated apoptosis. The signaling pathway for Fas-mediated apoptosis in various cells, including HL-60 cells, is currently unknown. Here, we studied the role of survival/apoptosis associated phosphatidylinositol 3-kinase (PI 3-K)/Akt in this process. We found that both PI 3-K inhibitors, wortmannin and LY294002, markedly suppressed phosphorylation of Akt and Bad in HL-60 cells. PI 3-K inhibitors significantly accelerated not only spontaneous apoptosis, but also Fas-induced apoptosis in HL 60 cells. The pro-apoptotic effect of PI 3-K inhibitors favored Fas-mediated apoptosis rather than spontaneous apoptosis in HL-60 cells. The caspase-3 or -8 inhibitor reduced the pro-apoptotic effect of the PI 3-K inhibitors for Fas mediated apoptosis in HL-60 cells, but the caspase-9 inhibitor did not. Although PI 3-K inhibitors did not affect Fas expression in HL-60 cells, cellular FLICE inhibitory protein (c-FLIP) levels were markedly reduced by PI 3-K inhibitor treatment. Furthermore, antisense oligonucleotide of c-FLIP confirmed that down regulation of c-FLIP enhanced sensitization to Fas-mediated apoptosis in HL-60 cells. These results suggest that the PI 3-K/Akt signaling pathway may, in part, regulate Fas-mediated apoptosis in HL-60 cells through c-FLIP expression. PMID- 16211289 TI - Immunohistochemical demonstration of fluoropyrimidine-metabolizing enzymes in various types of cancer. AB - Fluoropyrimidines [5-Fluorouracil (5-FU) and its prodrugs] have been widely used in the treatment of solid cancers. The anticancer effects primarily depend on intratumoral levels of enzymes metabolizing the drugs, such as dihydropyrimidine dehydrogenase (DPD), orotate phosphoribosyltransferase (OPRT), thymidine phosphorylase (TP), and thymidylate synthase (TS). In order to know the tumor types susceptible to respective fluoropyrimidines, we investigated the expression of DPD, OPRT, TP and TS in various types of cancer with the immunoperoxidase method. These four enzymes existed in all of the cancer types studied, such as pulmonary, gastric, colorectal, hepatic, cholecystic, pancreatic, renal, urocystic, and mammary cancers. Respective types of cancers presented characteristic immunohistochemical features as follows: pulmonary adenocarcinoma, DPD- and TP-high; pulmonary squamous cell carcinoma, TS- and TP-high; intestinal type gastric adenocarcinoma, TP-high; diffuse-type gastric adenocarcinoma, DPD low and TS-high; colorectal adenocarcinoma, DPD- and TP-low, hepatocellular carcinoma, DPD-high, and TS- and OPRT-low; cholecystic adenocarcinoma, DPD- and TS-high; renal cell carcinoma, DPD-low, and OPRT- and TP-high; urocystic transitional cell carcinoma, DPD-high and OPRT-low; and mammary ductal carcinoma, OPRT-low, and TS- and TP-high. The enzyme expression pattern in cancer tissue was generally similar to that of their normal counterparts. However, TP immunoreactivity in adenocarcinomas of the lung, stomach and gallbladder, and urothelial carcinoma of the urinary bladder was stronger, and DPD immunoreactivity in adenocarcinoma of the breast was weaker, when compared with normal epithelial cells. Non-epithelial cells were also positive for these enzymes. These results indicated that the key enzymes influencing the effects of fluoropyrimidines differ from cancer to cancer. Fluoropyrimidine treatment may be selected, based on the simultaneous immunohistochemical evaluation of the fluoropyrimidine metabolic enzymes. PMID- 16211290 TI - Classification of neural differentiation-associated genes in P19 embryonal carcinoma cells by their expression patterns induced after cell aggregation and/or retinoic acid treatment. AB - Expression of neural differentiation-associated genes was examined by RT-PCR and macroarray analyses during neural differentiation of P19 embryonal carcinoma cells induced by cell aggregation and/or retinoic acid (RA) treatment. Results revealed that the neural genes examined could be classified into 4 groups based on their expression patterns. The 1st group included the Wnt-1, Id-1, Id-3 and cdc42 genes, expression of which was altered by cell aggregation alone, but not by RA treatment alone. The 2nd group included the alphaN-catenin, Neuro D and GDNFRbeta genes, expression of which was altered by RA treatment alone, but not by cell aggregation. The 3rd group consisted of the Brn-2, TrkA, bcl-X, N cadherin, E-cadherin and Otx-2 genes, expression of which was altered by either treatment. The 4th group included the ACTH, D4DR, NGC and Oct-3 genes, the expression of which changed only when both treatments were applied simultaneously. Expression of the Ets-1 and Fli-1 transcription factor genes was up-regulated by either treatment alone at initial stages of neural differentiation of P19 cells, although overexpression of these genes alone could not induce cell differentiation. Our results suggest that although both treatments are required for complete neural differentiation of P19 cells, cell aggregation or RA treatment alone drive differentiation to a certain extent at the gene expression level. PMID- 16211291 TI - Prognostic value of human papillomavirus in the survival of head and neck cancer patients: an overview of the evidence. AB - Despite improvements over the last decade, survival rates for head and neck squamous cell carcinoma (HNSCC) remain around 50% and variable. Recently, a subset of HNSCC has been shown to contain human papillomavirus (HPV) and the presence of HPV in tumors constitutes a prognostic marker of disease. HPV-DNA in these tumors (overwhelmingly genotype 16, a high-risk type also found in cervical cancer) is present at high copy numbers, frequently integrated, and often transcriptionally active. In comparison with HPV-negative tumors, however, HPV positive HNSCCs are more likely to be located within the oral cavity/pharynx and diagnosed at a late stage. Based on the epidemiological evidence to date, it is reasonable to hypothesize that HPV-positive and -negative HNSCC represent different lineages formed through diverse, though overlapping, mechanisms of multistage tumorigenesis. The introduction of prophylactic and therapeutic HPV vaccines could have important implications for the prevention and control of a substantial fraction of HNSCC. PMID- 16211292 TI - Caspase-8 is scarcely silenced and its activity is well correlated with the anticancer effect of tumor necrosis factor-related apoptosis-inducing ligand in gastric cancer cells. AB - The main apoptotic signal stimulated by tumor necrosis factor-related apoptosis inducing ligand (TRAIL) streams through caspase-8 activation and evokes caspase 3, a central apoptosis activator. In this study, the status of caspase-8 and -3 in gastric cancer cells related to the anticancer effects of TRAIL was investigated. In the caspase-8 gene promoter, 9 of 10 gastric cancer cell lines harbor no hypermethylation. The pretreatment amounts of caspase-8 and -3 in these cells were not predictors for the anticancer effect of TRAIL. Caspase-8 activity 24 h after treatment with TRAIL was well correlated with the anticancer effect of TRAIL (r=0.777, p=0.0060). Caspase-3 activity 24 h after treatment with TRAIL showed a trend towards an association with the anticancer effect of TRAIL (r=0.544, p=0.1067). These results suggested that gastric cancer might be a good target of TRAIL therapy because the majority of tumor cells have intact caspase-8 expression. The anticancer efficacy may be predicted by the degree of caspase-8 activation after TRAIL treatment. PMID- 16211293 TI - Analysis of local recurrence after breast conservative treatment for invasive breast cancer: a single institution cohort. AB - In a group of 900 patients treated for carcinoma of the breast, we evaluated patient-, tumour- and treatment-related parameters, predicting the success or failure of conservative treatment for invasive breast cancer. Thirty-one patients developed local recurrences which were detected within 0.1-12.1 years after treatment of the primary tumour (with a median of 6.2 years), providing a risk of 2% at 5 years and 9% at 10 years. The locally recurrent tumours and their original primary tumours of 28 patients could be retrieved from the pathology laboratory archives. These 28 tumours of the recurrence group (RG) were matched with tumours without local recurrence, the non-recurrence group, for age at time of diagnosis, duration of follow-up and T- and N-stage. The tumours were studied for type and grade of invasive tumour including the in situ component and involvement of surgical margins. In addition, the expression of cell-cycle proteins, p53, Ki-67 (MIB-1) and BCL-1, as well as HER-2/ neu oncoprotein, estrogen and progesterone receptor were investigated. We found a mean age at diagnosis for the RG of 50 years, and the mean age at time of diagnosis for the whole group of 900 patients was 56 years (p=0.003). Thirty-nine percent of the RG had a positive surgical margin, which was the case for only 18% in the control group (p=0.09). The presence of the in situ component was also correlated with increased local recurrence (p=0.022). Furthermore, local recurrence was also associated with a significantly increased occurrence of distant metastases (p=0.001). We conclude that breast conservative treatment is safe with a low local recurrence rate. PMID- 16211294 TI - Identifying genes with differential expression in gemcitabine-resistant pancreatic cancer cells using comprehensive transcriptome analysis. AB - Pancreatic cancer is often unresectable at diagnosis, and chemotherapy using gemcitabine is now the standard treatment for advanced pancreatic cancer. However, acquired resistance to gemcitabine resulting in therapeutic failure is often encountered. Therefore, we sought to identify genes that determine gemcitabine resistance by evaluating the relationship between gene expression profiles and gemcitabine sensitivity to provide molecular targets for overcoming gemcitabine resistance. First, the gemcitabine concentration needed for 50% growth inhibition was examined in six pancreatic cancer cell lines. By exposing MIA PaCa-2 cells to long-term gemcitabine, we established gemcitabine-resistant cells. The gene expression profiles of the six pancreatic cancer cell lines and gemcitabine-resistant cells were determined using cDNA microarray analysis. By comparing the results, 30 genes were identified as differentially expressed genes correlated with gemcitabine sensitivity. Differentially expressed genes in the parental cell lines were also examined, and six overlapping genes were identified as genes correlated with gemcitabine sensitivity in both assays. Of these genes, the down-regulated expression of TNFSF6 protein, also known as Fas ligand, was confirmed in the gemcitabine-resistant cell line. These results should provide therapeutic molecular targets for overcoming gemcitabine resistance. PMID- 16211295 TI - CD4+CD25+ regulatory T cells in the peripheral blood of patients with breast cancer and non-small cell lung cancer. AB - It is well known that immunosuppression may contribute to the progression and chemotherapy-resistance of cancer. Recent studies have demonstrated that lymphocytes with the phenotype of CD4+CD25+ regulatory T cells (T-regs) contribute to immune dysfunction in cancer patients, and a relative increase in CD4+CD25+ regulatory T cells is related to immunosuppression and tumor progression in patients with some malignancies. In the present study, we evaluated the prevalence of T-regs in the peripheral blood of patients with breast cancer and non-small cell lung cancer. The phenotype of lymphocyte CD4+CD25+ cells was analyzed in peripheral blood of patients with breast cancer (n=22) and non-small cell lung cancer (NSCLC) (n=17). The population of CD4+CD25+ cells in CD3+ and CD4+ cells was evaluated by flow cytometric analysis with triple-color staining. Patients with breast cancer did not have a higher percentage of CD4+CD25+ cells in the total CD3+ and CD4+ cells in their peripheral blood than healthy volunteers. In contrast, patients with recurrent NSCLC had significantly higher percentages of CD4+CD25+ cells in CD3+ (47.6%) and CD4+ (71.0%) than healthy volunteers (n=10) who had CD4+CD25+ cells in CD3+ (33.7%, p=0.02) and CD4+ (52.2%, p<0.03). The population of CD4+CD25+ T-regs in the peripheral blood of patients with non-small cell lung cancer was significantly higher than that in healthy volunteers but not in breast cancer patients. These findings suggest that the use of T-reg-targeted immunomodulatory therapy may be a more effective strategy for patients with non-small cell lung cancer than for those with breast cancer. PMID- 16211296 TI - Down-regulation of O6-methylguanine-DNA methyltransferase gene expression in gliomas by platinum compounds. AB - Forty-two patients with malignant gliomas that had received two courses of chemotherapy more than 2 months apart were examined. Among these 42 patients, 31 were treated with 1-(4-amino-2-methyl-5-pyrimidynyl) methyl-3-(2-chloroethyl)-3 nitrosourea hydrochloride (ACNU), and 11 were treated with platinum compounds such as cis-platinum (CDDP) or carboplatin (CBDCA), as the first-line chemotherapy. The response rate of the second chemotherapy in the 31 patients treated first with ACNU was significantly lower than that in the 11 patients treated with platinum compounds, regardless of the type of the second chemotherapy (P=0.0292 by Fisher's exact probability test). O(6)-methylguanine DNA methyltransferase (MGMT) mRNA expression was measured twice by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) using Sybr Green I in 16 of 42 patients. The relative quantitation value (RQV) of MGMT mRNA normalized to the level of 2-microglobulin decreased after chemotherapy in all 5 patients treated with platinum compound. U373MG and A172 human glioma cells were cultured for 5 days with 1 microM of CDDP or 4 microM of CBDCA. The RQV of MGMT in these cells treated with platinum compounds obviously decreased, and these cells were more sensitive to ACNU than the control cells based on colorimetric 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Both the clinical findings and laboratory results suggest that platinum compounds may play a role in the down-regulation of MGMT mRNA expression and up-regulation of the sensitivity to ACNU. Platinum compounds may be strong candidates for use as first line chemotherapeutic agents against malignant gliomas. PMID- 16211297 TI - Establishment and characterization of a cisplatin-resistant oral squamous cell carcinoma cell line, H-1R. AB - Cisplatin (CDDP) is a widely used potent chemotherapeutic agent for many malignancies. However, the mechanism of resistance to CDDP remains unclear. To investigate the molecular mechanism, we established a CDDP-resistant cell line (H 1R) from a CDDP-sensitive cell line (H-1) which was derived from moderately differentiated squamous cell carcinoma of the lower gingiva. The 3-(3,4-dimethyl thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay indicated that H-1R had a 10-fold greater resistance to CDDP than H-1. When we compared gene expression levels in the cell lines using an in-house cDNA microarray, which represented 2,201 genes originating from normal oral tissue, primary oral cancer, and oral cancer cell lines, 12 genes showing elevated mRNA expression in H-1R compared with H-1 were identified. Among them, the up-regulated expression of ATP-binding cassette transporter genes (MDR1, MRP1, and MRP2), CD55, and PGK1 and down regulated expression of Caveolin 1 were further confirmed by semiquantitative reverse transcriptase-polymerase chain reaction (PCR) or real-time PCR. Our results suggest that H-1 and H-1R cell lines could be useful for elucidating the candidate genes responsible for CDDP resistance, including the genes found in this study. PMID- 16211298 TI - C2-ceramide exhibits antiproliferative activity and potently induces apoptosis in endometrial carcinoma. AB - The purpose of the present study was to investigate the effects of an exogenously administered cell-permeable synthetic ceramide analogue, C(2)-ceramide (N-acetyl sphingosine) on the growth, cell cycle, and death of Ishikawa human endometrial carcinoma cells. We investigated the effects of C(2)-ceramide on Ishikawa endometrial cancer cell lines in vitro. The cells were treated with C(2) ceramide, and its effects on cell growth, cell cycle, apoptosis, and related measurements were investigated. MTT assays showed that C(2)-ceramide, a cell permeable analogue of ceramide, significantly induced dose- and time-dependent death in human endometrial carcinoma Ishikawa cells. Cell-cycle analysis indicated that their exposure to C(2)-ceramide decreased the proportion of cells in S phase and increased the proportion in G0/G1 and/or G2/M phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis, including cleavage of poly-ADP ribose polymerase. Furthermore, we demonstrated that the amount of phosphorylated Akt was decreased by C(2)-ceramide. These results raise the possibility that C(2) ceramide may prove particularly effective in the treatment of endometrial cancers. PMID- 16211299 TI - Identification of candidate radioresistant genes in human squamous cell carcinoma cells through gene expression analysis using DNA microarrays. AB - Radiation therapy is currently the standard adjuvant approach for oral squamous cell carcinoma (OSCC) patients. Individual OSCCs display a wide range of radiosensitivity (RS). To identify genes associated with radioresistance (RR) of OSCC and establish a useful method of predicting radio-therapeutic effectiveness, we examined the gene expression patterns of OSCC cell lines that exhibited different responses to ionizing radiation (IR) by clonogenic survival assay using an in-house cDNA microarray consisting of 2,201 human genes and quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR). Microarray analysis showed overexpression of 7 genes in the radioresistant cell line, HSC2, and 2 genes in the radiosensitive cell line, HSC3. The changes in expression levels in 7 of 9 genes (Cytokeratin18, DTNBP1, ASNA1, Tcp20, Cyclophilin F, KIAA0218, and HBp17) were confirmed with QRT-PCR. Of these, the genetic alterations of Tcp20, whose expression was remarkably elevated in radioresistant HSC2 cells after IR, were investigated. The escalation of X-ray doses resulted in an enhanced Tcp20 expression level in HSC2 cells compared to radiosensitive HSC3 cells (P<0.05, Mann-Whitney U test). These results suggest that the identified genes, which include Tcp20, may play an important role in conferring RR to OSCC, and could also be useful in identifying cases of OSCC with more radioresistance. PMID- 16211300 TI - Quercetin induces gadd45 expression through a p53-independent pathway. AB - Quercetin, a kind of flavonoid, is found in edible fruits and vegetables and has anti-tumorigenic activity. However, the mechanism of activity has not been elucidated. We show for the first time that gadd45 is a molecular target of quercetin, which inhibits growth of human cervical cancer HeLa cells. Apoptosis was detected in HeLa cells treated with quercetin. At the concentration inducing apoptosis, quercetin also increased gadd45 expression at the mRNA and protein level, however, the 5'-promoter region of the gadd45 gene was not activated by quercetin. Since gadd45 is known to be a downstream gene of the tumor suppressor p53, we examined whether or not quercetin regulates gadd45 induction via a p53 pathway. Quercetin did not activate transcription through p53-binding sites in HeLa cells, although it up-regulated gadd45 in p53-inactivated tumor cells. These results indicate that quercetin induces gadd45 expression in a p53-independent manner. PMID- 16211301 TI - Mitochondria-independent induction of Fas-mediated apoptosis by MSSP. AB - Fas-mediated apoptosis has been proposed to play an important role in homeostasis. Fas triggers apoptosis after stimulation by its ligand FasL or the Fas ligand agonist anti-Fas antibody through a mitochondria-dependent or independent pathway, and MSSP has been identified as a transcription factor that regulates the c-myc gene and was later found to positively or negatively regulate a variety of genes, including alpha-smooth actin, MHC class I, MHC class 2 and the thyrotropin receptor. We further found that expression of the Fas gene was repressed, resulting in abrogation of the Fas-mediated induction of apoptosis both in Mssp-knockout mice and primary thymocytes. MSSP was then found to stimulate promoter activity of the Fas gene by binding to a specific region. In this study, to identify the MSSP-dependent Fas-induced apoptosis pathway, primary fibroblasts from MSSP (+/+) and MSSP (-/-) cells were treated with the combination of interleukin 1-beta and interferon-gamma and expression of the Fas gene was examined. The results showed that the Fas gene was expressed at the same levels in the two cell types. Furthermore, when these cells were treated with the anti-Fas antibody, it was found that cytochrome C was not released in the cytosol and that activations of caspase 8 and caspase 3 occurred in primary fibroblasts from MSSP (+/+) cells but not from MSSP (-/-) cells. These results indicate that Fas-mediated apoptosis induced by MSSP occurs independently of mitochondria. PMID- 16211302 TI - Targeting of X-linked inhibitor of apoptosis protein or survivin by short interfering RNAs sensitize hepatoma cells to TNF-related apoptosis-inducing ligand- and chemotherapeutic agent-induced cell death. AB - The inhibitors of apoptosis (IAPs) family regulate apoptosis by preventing the action of the central execution phase, and function as mediators and regulators of the anti-apoptotic activity of the v-Rel and NF-kappaB transcription factor families. The targeting of IAPs may be a promising strategy, but it is not well elucidated in human hepatocellular carcinomas (HCCs). We have therefore investigated the effects of the down-regulation of IAPs (XIAP or survivin) on the TNF-related apoptosis-inducing ligand (TRAIL) and chemotherapeutic agents that induced apoptosis in human HCC cells. To inhibit the IAPs gene expression, we designed small interfering RNA (siRNA) against the X-chromosome-linked IAP (XIAP) or survivin and investigated their efficacy in the suppression of the XIAP or survivin expression in two HCC cells (SK-Hep1 and HLE), and their consequent antitumor potential. We found that the designed siRNAs against the XIAP and survivin downregulated the protein expression of respective genes by almost 50%. The suppression of IAPs resulted in a significant decrease in procaspase-3 levels, especially by suppression of the XIAP. The apoptosis cell count was small in cells transfected with control siRNA and siRNA against the XIAP or survivin, but after treatment with 10 ng/ml of TRAIL, the apoptosis cells increased 2-3 times by the suppression of IAPs as control. The cytotoxicity of doxorubicin and camptothecin was augmented by the suppression of the XIAP in SK-Hep1 cells, whereas the suppression of survivin did not affect cytotoxicity. In conclusion, downregulation of the XIAP or survivin enhances cell death by TRAIL and increases sensitivity against some chemotherapeutic agents in HCC cells. In particular, the XIAP may be a potential target to increase therapeutic sensitivity. PMID- 16211303 TI - Effects of tamoxifen and soluble tumor-associated antigens on ovarian structure in mammary tumor-bearing rats. AB - Previously, we showed that the 66 and 51 kDa soluble tumor-associated antigens (sTAAs) have distinct suppressive effects on chemically induced mammary cancer in rats, both alone and in combination with the hormone-related anticancer drug tamoxifen. Here, we describe the effects of both sTAA and tamoxifen on the histological structure of ovaries in mammary tumor-bearing 30- to 34-week-old rats. Central ovary sections were pooled, the number of the healthy and degenerated follicles were counted, and the size of the corpora lutea was estimated. In follicular development primordial, primary, preantral and antral stages were recognized. Only healthy follicles with visible nuclei were counted. Follicular degeneration was estimated as the number of atretic follicles with follicular remnants. Treatment with tamoxifen alone or in combination with sTAA significantly increased the number of primordial follicles and atretic follicles in the ovaries, and promoted the formation of small follicular cysts. Total area of the corpora lutea decreased. sTAA participated in this process by increasing apoptosis in degenerated follicles. PMID- 16211304 TI - Identification of reproducible low mass SELDI protein profiles specific to cisplatin resistance in human ovarian cancer cells. AB - Platinum compounds are the most effective drugs in the fight against ovarian cancer. Unfortunately, many ovarian tumors are not eradicated by chemotherapy due to the emergence of drug-resistant clones during therapy, and hence 5-year survival rate of women afflicted with this disease is just 18%. In the continued absence of an effective early detection test for ovarian cancer, there is a considerable need to develop treatment strategies that can either circumvent (e.g. gene therapy) or prevent the development of platinum resistance. A prerequisite for the development of such treatments is a detailed knowledge of factors that confer tumor cell resistance to platinum compounds. We have used surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS or SELDI) to identify low mass proteins that are uniquely expressed in ovarian tumor cells that are platinum-resistant. Only two polypeptide peaks (m/z 5041 and 7324) were consistently altered following the induction of cisplatin resistance in the OAW42 and 2780 cell lines. These peaks appear to be specific to cisplatin resistance as they are not altered in the same manner in a melphalan-resistant variant of OAW42. The exact identity of the polypeptide peaks is unknown, but appears to be unrelated to changes in several proteins that have been historically associated with cisplatin resistance. These data suggest that SELDI-based proteomic profiling may be useful in monitoring the emergence of cisplatin-resistant tumor cell clones. PMID- 16211305 TI - In vitro studies of the dry fruit of Chinese fan palm Livistona chinensis. AB - The hot water extract of the dry fruit of Chinese fan palm Livistona chinensis R Br. (LC) has been used in folklore medicine in Southern China for treating various tumors. Our in vitro study showed that the ethanolic extract (LCET) and the hot water extract (LCWE) of LC inhibited HL60 cell growth, with 50% inhibition (IC(50)) estimated at a 1/50 dilution for both preparations. LCET showed mild activity in inducing HL60 cell differentiation into granulocyte lineage. However, at 1/100 and 1/200 dilutions, it respectively induced 32.4+/ 12.6% and 16.3+/-6.1% of HL60 cells into monocyte/macrophage lineage, compared to 4.4+/-1.3% in the control. In contrast, LCWE did not demonstrate a significant differentiation-inducing capacity on HL60 cells. Cell-free Trolox equivalent antioxidant capacity hydroxyl radical scavenging assay estimated that a 1/10 dilution of LCET and LCWE has a similar activity, equivalent to 13.0 and 12.7 microM of Trolox activity respectively. At a 1/100 dilution, neither extract affected nitric oxide production in both non-stimulated and lipopolysaccharide (LPS) stimulated RAW 264.7 cells. RT-PCR analyses of mRNA expression showed that treatment of RAW 264.7 cells with either extract at a 1/100 dilution did not affect tumor necrosis factor-alpha (TNFalpha), interleukin-1beta (IL-1beta), inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) mRNA expression in these cells compared to the untreated control. Neither extract affected TNFalpha and iNOS mRNA expression in LPS-stimulated cells, but at a 1/100 dilution they both reduced IL-1beta mRNA expression in LPS-stimulated cells (p<0.01). Only a 1/100 dilution of LCET reduced COX-2 mRNA expression in LPS stimulated RAW 264.7 cells (p<0.01). The dry fruit of Livistona chinensis warrants further investigation. PMID- 16211306 TI - Targeted adriamycin delivery to MXT-B2 metastatic mammary carcinoma cells by transferrin liposomes: effect of adriamycin ADR-to-lipid ratio. AB - In the protein-targeted therapy for cancer, transferrin (Tf) is used to reach a selective and specific target in cancer cells. Tf is used conjugated to chemotherapeutic drugs, insulin, toxins, antibodies, polymers, nanoparticles, lipoplexes and liposomes. Using this latter approach, hydrophobically derivatized Tf was incorporated to liposomal bilayers. The biological activity of Tf-liposome was tested using MXT-B2 cells, a metastatic mammary carcinoma cell line. In Tf binding assays, the Scatchard analysis indicated 4.5x10(5) Tf receptors/cell. In cell growth assays, Tf-liposomes stimulated cell growth in a dose-dependent manner, up to a maximum of 32% of the total free Tf stimulation. Following this, we prepared Tf-liposomes encapsulating adriamycin (ADR) at two different ADR-to lipid ratios. In vitro cytotoxicity assays against MXT-B2 cells gave IC(50) values 2.1-times lower for Tf-liposomal ADR in comparison to control liposomal ADR. However, similar IC(50) values were found for low ADR-to-lipid ratio Tf liposomal ADR, as well as for control liposomal ADR. The free Tf added in excess increased the IC(50) value of Tf-liposomal ADR by 51%, while the IC(50) value of control liposomal ADR was unaffected, supporting a receptor-mediated mechanism of targeting by Tf. In addition, the lower IC(50) value is correlated with a higher total of ADR accumulation in the cells. PMID- 16211307 TI - Antioxidant and anti-inflammatory properties of ESSIAC and Flor-Essence. AB - Essiac (ES) and Flor-Essence (FE) are two herbal teas widely taken by North American cancer patients during chemo- and radiation-therapy. The antioxidant and anti-inflammatory properties of these two herbal teas were assessed in this study. Cell-free Trolox equivalent antioxidant capacity assay shows that at 1/5 dilution, ES and FE have hydroxyl radical scavenging activity equivalent to 10.65 microM and 5.74 microM of Trolox respectively. Treatment with ES at 1/10 and 1/20 dilutions significantly increased nitric oxide (NO) production by murine RAW 264.7 cells, but inhibited NO production in a concentration-dependent manner by lipopolysaccharide (LPS)-stimulated cells. In contrast, FE at 1/10 and 1/20 dilutions did not significantly induce NO production by RAW 264.7 cells, nor did it, at these dilutions, inhibit the NO production by LPS-stimulated RAW 264.7 cells. RT-PCR assay shows that both 1/20 and 1/100 dilutions of ES and FE induced mRNA expression of IL-1beta, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) pro-inflammatory molecules in RAW 264.7 cells compared to untreated controls. Addition of ES and FE at 1/20 and 1/100 dilutions to LPS stimulated RAW 264.7 cells did not alter IL-1beta, iNOS and COX-2 mRNA expression in these cells. ES and FE treatment did not affect TNFalpha mRNA expression in non-stimulated or LPS-stimulated RAW 264.7 cells. Our data show that ES but not FE stimulated NO release by non-stimulated and inhibited LPS-stimulated RAW 264.7 cells. There were only minor differences between ES and FE in their induction of mRNA expression of pro-inflammatory molecules. Further research is needed to investigate the differential activities of these two herbal teas in stimulating pro-inflammatory mediators release by RAW 264.7 cells. PMID- 16211308 TI - Comparative genomics on SLIT1, SLIT2, and SLIT3 orthologs. AB - SLIT1 gene at human chromosome 10q24.1, SLIT2 gene at 4p15.31, and SLIT3 gene at 5q34-q35.1 encode large secreted proteins functioning as ligands for Roundabout (Robo) receptors. SLIT-ROBO signaling pathway is implicated in neurogenesis, angiogenesis, and immune response through the regulation of axonal guidance, endothelial cell migration, and denderitic cell migration, respectively. GREMLIN (CKTSF1B1 or GREM1) and DANTE (CKTSF1B3 or GREM3) are secreted antagonists for BMPs and SLITs. Here, comparative integromics analyses on SLIT1, SLIT2, and SLIT3 orthologs were performed by using bioinformatics. Rat Slit2 gene, consisting of 36 exons, was located within rat genome sequences AC098362.4 and AC111627.6. Mouse Slit3 complete coding sequence was determined by assembling BB634238 EST, AF144629 cDNA, and AK129223 cDNA. Leucine-rich repeats with nine conserved cysteine (LRRCC) domains and SLIT C-terminal cysteine-rich (SLITCCR) domain were identified in this study. CPxxCxCxxxxVxCxxxxLxxxPxxxPx(10~58) Nx(19,20)LxxNx(9)Fx(8)LxLxxNxxxCxxxxxFxxLxxx xxLxLxxNx(9)Fx(13)NxxxCxCxxxWLx(15)CxxPx(17)C was the consensus sequence of LRRCC domain. Mammalian SLIT1, SLIT2 and SLIT3 orthologs were large secreted proteins with four LRRCC domains, nine EGF domains, Laminin G (LamG) domain, and SLITCCR domain. SLIT1 mRNA was expressed in fetal brain, infant brain, anaplastic oligodendroglioma, and Jurkat T cells. SLIT2 and SLIT3 mRNAs were co-expressed in embryonic stem (ES) cells with embryoid body formation, and diffuse type gastric cancer with signet ring cell features. Double TCF/LEF and bHLH-binding sites were conserved among mammalian SLIT1 promoters. FOXJ2, E47, ETS1, and FOXA2-binding sites and CCAAT box were conserved among mammalian SLIT3 promoters. Mammalian SLIT1 orthologs were identified as evolutionarily conserved targets of the WNT/beta-catenin signaling pathway. PMID- 16211309 TI - The pyrrolo-1,5-benzoxazepine, PBOX-6, inhibits the growth of breast cancer cells in vitro independent of estrogen receptor status and inhibits breast tumour growth in vivo. AB - Members of a novel series of pyrrolo-1,5-benzoxazepine (PBOX) compounds have been shown to induce apoptosis in a number of human leukemia cell lines of different haematological lineage, suggesting their potential as anti-cancer agents. In this study, we sought to determine if PBOX-6, a well characterised member of the PBOX series of compounds, is also an effective inhibitor of breast cancer growth. Two estrogen receptor (ER)-positive (MCF-7 and T-47-D) and two ER-negative (MDA-MB 231 and SK-BR-3) cell lines were examined. The 3,4,5-dimethylthiazol-2-yl-2,5 diphenyl-tetrazolium bromide (MTT) assay was used to determine reduction in cell viability. PBOX-6 reduced the cell viability of all four cell lines tested, regardless of ER status, with IC(50) values ranging from 1.0 to 2.3 microM. PBOX 6 was most effective in the SK-BR-3 cells, which express high endogenous levels of the HER-2 oncogene. Overexpression of the HER-2 oncogene has been associated with aggressive disease and resistance to chemotherapy. The mechanism of PBOX-6 induced cell death was due to apoptosis, as indicated by the increased proportion of cells in the pre-G1 peak and poly(ADP-ribose) polymerase (PARP) cleavage. Moreover, intratumoural administration of PBOX-6 (7.5 mg/kg) significantly inhibited tumour growth in vivo in a mouse mammary carcinoma model (p=0.04, n=5, Student's t-test). Thus, PBOX-6 could be a promising anti-cancer agent for both hormone-dependent and -independent breast cancers. PMID- 16211310 TI - Combination effects of irradiation and irinotecan on cervical squamous cell carcinoma cells in vitro. AB - Irinotecan HCl (CPT-11) has frequently been used in chemotherapy or concurrent chemoradiotherapy for patients with advanced cervical cancer, although an effective protocol for chemoradiotherapy with CPT-11 has not yet been established. Using the radiosensitive human cervical squamous cell carcinoma cell line ME180 and SN38, a major active metabolite of CPT-11, both the SN38 effects on radiosensitivity and irradiation effects on SN38 sensitivity were investigated to optimize the chemoradiotherapy protocol for CPT-11. SN38 had no effect on radiosensitivity, and irradiation did not affect SN38 sensitivity. Moreover, 3 of 4 post-irradiation surviving subclones obtained from repeatedly irradiated ME180 cells showed no significant changes in their SN38 sensitivities compared with the non-irradiated parent cells. On the other hand, all 7 SN38-resistant subclones established from ME180 cells showed strong reduction in their radiosensitivities. These results suggest that CPT-11 should be administered to advanced cervical cancer patients after, but not before, standard radiotherapy, and concurrent administration of SN38 with radiotherapy should be avoided in order to prevent severe adverse effects, such as watery diarrhea, which is the main adverse effect of pelvic radiotherapy or CPT-11. PMID- 16211311 TI - Enhanced expression of the secreted form of clusterin following neoadjuvant hormonal therapy as a prognostic predictor in patients undergoing radical prostatectomy for prostate cancer. AB - The objective of this study was to characterize changes in clusterin expression in prostate cancer before and after neoadjuvant hormonal therapy (NHT), and to assess the prognostic significance of clusterin expression following NHT in patients undergoing radical prostatectomy. Paired needle biopsy and corresponding radical prostatectomy (RP) specimens obtained from 76 patients were analyzed for expression of clusterin protein by immunohistochemical staining with an antibody recognizing all of the clusterin isoforms. RP specimens following NHT demonstrated stronger expression of clusterin than the corresponding needle biopsy specimens. Clusterin protein was present in the cytoplasm of both biopsy and RP specimens; however, there was no nuclear staining identified in any specimens. The expression level of clusterin in biopsy specimens was significantly associated with the biopsy Gleason score, but not with other parameters available before RP. Furthermore, clusterin expression in RP specimens was significantly associated with the preoperative prostate specific antigen value and pathological stage, but not with other pathological factors. Biochemical recurrence-free survival in patients with strong clusterin expression in RP specimens was significantly lower than that in those with weak clusterin expression; however, the level of clusterin expression could not be used as an independent predictor of biochemical recurrence. These findings suggest that despite the lack of independent significance, the expression level of the secreted form of clusterin in prostate cancer tissue after NHT, which may inversely reflect the therapeutic effect of NHT, could be a useful parameter predicting biochemical recurrence in patients undergoing RP. PMID- 16211312 TI - Minimal residual disease as the target for immunotherapy and gene therapy of cancer (review). AB - Local recurrences at the site of tumour resection as well as distant micrometastases manifested after surgery represent major problems in oncology. Adjuvant immunotherapy and gene therapy may help to cope, at least partially, with these problems. Adjuvant modalities may be more effective in treating residual tumour disease compared to bulky tumours, owing to a favourable effector/target cell ratio. The purpose of this review was to summarize, evaluate and discuss the results obtained with adjuvant immunotherapy and immunomodulatory gene therapy of surgical minimal residual tumour disease in experimental and clinical tumour systems. The prospects and limitations of adjuvant therapeutic modalities will be considered. PMID- 16211313 TI - Comparative genomics on ROR1 and ROR2 orthologs. AB - Transmembrane proteins with extracellular Frizzled domain, such as ROR1, ROR2, MUSK, MFRP, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9 and FZD10, are key molecules for WNT signaling network. Here, comparative integromics analyses on ROR1 and ROR2 orthologs were performed by using bioinformatics. Zebrafish ror2 gene, consisting of nine exons, was identified within CR-450684.3 genome sequence. CV490605.1 EST corresponded to the 5'-end of zebrafish ror2 mRNA, and BM533602.1 EST corresponded to the 3'-end. Zebrafish ror2 gene was found to encode a 939-aa transmembrane protein, showing 71.7% and 56.2% total amino-acid identity with human ROR2 and ROR1, respectively. Immunoglobulin-like domain, Frizzled domain, Kringle domain within the extracellular region, tyrosine kinase domain, Ror homology C-terminal (RORHC) domain and juxta-C-terminal LLGD motif within the cytoplasmic region were conserved among vertebrate ROR1 and ROR2 orthologs. SH2 binding site within the RORHC domain was conserved among vertebrate ROR2 orthologs, but not among vertebrate ROR1 orthologs. ROR1 mRNA was expressed in embryonic stem (ES) cells, infant brain, renal cancer, and colon cancer. ROR2 mRNA was expressed in parathyroid, testis, uterus, and also in diffuse type gastric cancer with signet ring cell features. ROR2 promoter rather than ROR1 promoter was more evolutionarily conserved. WNT5A and ROR family receptors, co-expressed in ES cells and gastric cancer, are implicated in the planar cell polarity (PCP) pathway. ROR1 and ROR2 are the pharmacogenomics targets in the fields of stem cell biology and oncology. PMID- 16211314 TI - Identification of carotenoids in ovarian tissue in women. AB - Epidemiological and clinical studies have revealed that vitamin A and its derivatives (carotenoids and retinoids) can reduce the risk of ovarian tumours and may have a role in the metabolism of patients with ovarian cancer. The aim of the study was identification and quantitative assessment of carotenoids found in nature, mainly of provitamin A group, in the tissue material obtained from patients with different lesions of the ovaries. Material for analysis was obtained from 100 women, aged 16-74, operated on for ovarian tumours in the Department of Gynaecology. Carotenoid pigments were separated using column chromatography, thin-layer chromatography and high-performance liquid chromatography. In the tissue material subjected to analysis, 14 carotenoids were identified, including provitamin A carotenoids; beta-carotene, beta cryptoxanthin, echinenone and hydroxyechinenone. alpha-carotene was not found. In the whole group of pathological lesions, the total carotenoid content was relatively low (mean 1.717 microg/g tissue) and the mean content of provitamin A carotenoids was 17.28%. These results are similar to results obtained in the group of normal ovarian tissue. In the group of benign mucinous tumours (1.042 microg/g tissue) and tumours in the thecoma-fibroma group (1.328 microg/g tissue) and dysgerminoma group (1.279 microg/g tissue), the total carotenoid content was lower. Only in the endometriosis group was this value higher (2.185 microg/g tissue). Epoxy carotenoids; lutein epoxide, violaxanthin and mutatoxanthin were predominant (in %). Irrespective of histological classification, beta-carotene, beta-cryptoxanthin, lutein, lutein epoxide, violaxanthin and mutatoxanthin were identified in all tissue examined. Antheraxanthin was isolated in all tissue except for normal ovarian tissue, serous malignant and mucinous benign and malignant tumours, endometrioid malignant tumours, dermoid cysts, corpus luteum cysts and simple cysts. Hydroxyechinenone was isolated sporadically. Only in one case was capsanthin isolated. Carotenoids act as chemopreventive agents, irrespective of whether they are finally transformed into vitamin A, and may represent a potentially powerful alternative to present chemotherapeutic approaches to the treatment of ovarian cancer. PMID- 16211315 TI - Tension-free vaginal tape for the treatment of urodynamic stress incontinence with intrinsic sphincteric deficiency. AB - The purpose of this study was to evaluate the outcome of tension-free vaginal tape (TVT) procedure in women with urodynamic stress incontinence diagnosed as having intrinsic sphincteric deficiency (ISD). The combination of a maximal urethral closure pressure < 20 cm H2O and a Valsalva leak point pressure < 60 cm H2O was considered as diagnostic of ISD. Subjects with detrusor overactivity on preoperative urodynamics were excluded. A total of 35 patients with both low closure pressure and leak point pressure were enrolled. Bladder perforation occurred in three (8.6%) cases. Postoperative urinary voiding difficulties occurred in nine (25.7%) women. Two patients underwent surgical detension of the tape, with complete resolution of urinary retention and no relapse of incontinence. Women with postoperative voiding dysfunction had a significantly lower detrusorial pressure at the peak flow on preoperative urodynamics compared to those who voided efficiently after TVT. The mean (range) follow-up time was 12.5 months (3-36). The objective cure rate for stress incontinence was 91.4%. Two of the three (66%) patients in whom the TVT procedure failed had a fixed urethra. De novo urge incontinence was found in five (14.3%) patients. PMID- 16211316 TI - Anatomical study of the obturator foramen and dorsal nerve of the clitoris and their relationship to minimally invasive slings. AB - OBJECTIVES: The objective of the study is to clarify potential risks to the dorsal nerve of the clitoris (DNC) and obturator canal using different minimally invasive slings. STUDY DESIGN: Ten embalmed hemipelves were dissected to demonstrate the course of the DNC and the obturator canal. On each cadaver, tension-free vaginal tape (TVT), transobturator in-out (TVT-O) and transobturator out-in (Monarc) procedures were performed. Distances between the DNC and the obturator canal to the different devices were measured. RESULTS: The DNC passes beneath the pubic bone at a distance of 14.3 +/- 4.7 mm of the midline. The distances of the different devices to the DNC were similar. The distance to the obturator canal was significantly different, with TVT being the furthest (40.1 +/ 3.7 mm) and TVT-O the closest (19.3 +/- 3.1 mm; p < 0.0001). CONCLUSION: Given the course of the DNC along the medial aspect of the ischiopubic ramus, the out in technique may be safer. The in-out technique is the closest to the obturator canal. PMID- 16211317 TI - Unusual features associated with cranial openings of optic canal in dry adult human skulls. AB - One hundred and eighty six dry adult human skulls (372 sides) were studied in order to reveal the presence of unusual features--such as recess, fissure and notch--in the cranial opening of the optic canal. Recess, an extension of the lateral wall, was found in 229 (61.5%) sides. Fissure, an irregular discontinuity above the lateral recess, was observed in 158 (42.4%) sides. Notch, a gap in the posterior-most limit of the roof, was seen in 66 (17.7%) sides. These were commonly seen bilaterally. The etiology of these was attributed to the development of the optic canal. During intrauterine life the optic canal is in the form of a large keyhole shaped foramen, which slowly transforms into the adult canal. A developmental arrest at an early stage may lead to the formation of recesses and fissures. PMID- 16211318 TI - Enlarged perforating branch of peroneal artery and extra crural fascia in close relationship with the tibiofibular syndesmosis. AB - We found an extremely large perforating branch of peroneal artery in an 89-year old female cadaver's left ankle. The anterior tibial artery could not reach to supply the ankle and dorsum of the foot. The perforating branch of peroneal artery continued as the dorsalis pedis after giving off an anterior lateral malleolar artery branch. The posterior tibial artery was thinner than usual. On the anterior side of the ankle, there was an extra crural fascia in addition to the regular crural fascia, under the anterior crural muscles. This strong fascia was tightly overlying the perforating branch of peroneal artery and anterior tibiofibular ligament. It is important to know the relationship of these vessels to the surrounding structures. Surgeons must be careful while dissecting this area since the perforating branch of peroneal artery might be anomalously enlarged as well as crossing in front of the tibiofibular syndesmosis in order to prevent vascular injury. PMID- 16211319 TI - The arterial vascularization of the lateral tibial condyle: anatomy and surgical applications. AB - The contribution of the inferior lateral genicular artery (ILGA) and the anterior tibial recurrent artery (ATRA) in the arterial supply of the lateral tibial condyle (LTC) has not been comprehensively studied and remains controversial. Eleven knee joints were injected with colored latex and the arteries were dissected macroscopically. The ATRA yielded several osseous branches supplying the tibial metaphysis and the anterior part of the tibial epiphysis and several rami supplying the anterior tibial tuberosity and the lower part of the patellar tendon. The ILGA ran under the lateral collateral ligament and had a horizontal direction towards the retro-patellar fat pad. The ILGA yielded 4-6 branches ascending or descending perpendicularly to its main direction. Full anastomoses between branches derived from the ATRA and the ILGA were observed in front and behind the lateral intercondylar tubercle in all the specimens, but each vessel seemed to provide predominantly the blood supply to a specific area. The anterior part of the LTC drew its blood supply from the ATRA, the posterior part from the ILGA and the mid-portion from both arteries. The standard anterolateral approach to LTC fractures with sub-meniscal arthrotomy appears particularly harmful to epiphyseal vascularization since it interrupts many of the branches deriving from the ILGA and ATRA. The recent development of arthroscopy in the treatment of LTC fractures may be particularly advantageous as it spares the vascularization of the LTC. PMID- 16211320 TI - Multi-detector row CT scanning in Paleoanthropology at various tube current settings and scanning mode. AB - The purpose of this study was to determine the optimal tube current setting and scanning mode for hominid fossil skull scanning, using multi-detector row computed tomography (CT). Four fossil skulls (La Ferrassie 1, Abri Pataud 1, CroMagnon 2 and Cro-Magnon 3) were examined by using the CT scanner LightSpeed 16 (General Electric Medical Systems) with varying dose per section (160, 250, and 300 mAs) and scanning mode (helical and conventional). Image quality of two dimensional (2D) multiplanar reconstructions, three-dimensional (3D) reconstructions and native images was assessed by four reviewers using a four point grading scale. An ANOVA (analysis of variance) model was used to compare the mean score for each sequence and the overall mean score according to the levels of the scanning parameters. Compared with helical CT (mean score=12.03), the conventional technique showed sustained poor image quality (mean score=4.17). With the helical mode, we observed a better image quality at 300 mAs than at 160 in the 3D sequences (P=0.03). Whereas in native images, a reduction in the effective tube current induced no degradation in image quality (P=0.05). Our study suggests a standardized protocol for fossil scanning with a 16 x 0.625 detector configuration, a 10 mm beam collimation, a 0.562:1 acquisition mode, a 0.625/0.4 mm slice thickness/reconstruction interval, a pitch of 5.62, 120 kV and 300 mAs especially when a 3D study is required. PMID- 16211321 TI - Morphometry of the sacrum for clinical use. AB - Degenerative disease and instability in the lower lumbar spine may necessitate fusion and stabilization supplemented by instrumentation to the sacrum. However, screw placement in a reasonable position is more difficult to achieve because of the unique anatomy of the first sacral (S1) vertebra. Therefore, this study has been conducted to evaluate sacrum anatomy of the Western Anatolian population in terms of morphometric measurements and make a comparison with previous studies as well as giving a guidance to the surgeons. In this study, 60 dry adult sacrums (30 male and 30 female) were assessed for morphometric analysis. The measurement data for the sacrum and S1 vertebra revealed that there was no significant difference between both sexes except the sacral width and sacral canal width (p<0.05). In the present study, the ratio of S1 corpus' width to sacral width was lower in females compared with males. A detailed knowledge of the morphometric data about sacrum is very important for spinal surgery, as pedicle screw insertion is crucial in spinal instrumentation in order to prevent neurological injury and/or fixation failure. PMID- 16211322 TI - Study of the auditory tube by ventilation scintigraphy with technetium-99m. AB - The two essential regulating mechanisms of the middle ear pressure are the trans mucosal gas exchange in the middle ear and the ventilation function of the eustachian tube (ET). The physiological mechanism of these both functions is not yet clear. The purpose of this study was to evaluate the role of the ET pressure equilibrium function by ventilation scintigraphy with technetium-99m. The rabbit animal model in vivo was used to study the presence and role of the ventilation of the tympanic cavity via auditory tube. The obtained results did not show any ventilation function of the ET despite active opening by muscle movement. In our experience, ventilation scintigraphy with technetium-99m is not a reliable method to study the auditory tube pressure equilibrium function in physiological conditions. PMID- 16211323 TI - Proposed graphical system of evaluating disc-condyle displacements of the temporomandibular joint in MRI. AB - The aim of this preliminary study was to standardize the reading of magnetic resonance imaging (MRI) of the temporomandibular joint (TMJ). The MRI was conducted on a control group of eight subjects. In this study, the analysis of joint relationships was limited to the oblique sagittal plane, during the movement of controlled opening. The sections were analyzed by a computer-assisted quantitative method of graphical evaluation. The total surface area (TS) of the disc section was divided into an anterior surface area (AS) and a posterior area (PA), in accordance with a line joining the center of the condyle (C) to the center of the articular tubercle of the temporal bone (T). The disc-condyle relationships in the chosen plane of section were evaluated by the AS/TS ratio. If TS remains relatively constant, the AS/TS values on average decrease from 0.5 to 0.3 between the closed mouth position and the 25 mm open position. The results show good intra-observer reproducibility (p=0.26), evaluated by Wilcoxon's test. In conclusion, this protocol enables the displacement of the disc-condyle assembly during mouth opening to be described. Observed on a larger sample, the AS/TS would be able to show a range of variability in functional disc positions. PMID- 16211324 TI - Microbial diversity of the freshwater sponge Spongilla lacustris. AB - To provide insight into the phylogenetic bacterial diversity of the freshwater sponge Spongilla lacustris, a 16S rRNA gene libraries were constructed from sponge tissues and from lake water. Restriction fragment length polymorphism (RFLP) analysis of >190 freshwater sponge-derived clones resulted in six major restriction patterns, from which 45 clones were chosen for sequencing. The resulting sequences were affiliated with the Alpha proteobacteria (n=19), the Actinobacteria (n=15), the Beta proteobacteria (n=2), and the Chloroflexi (n=2) lineages. About half of the sequences belonged to previously described actinobacterial (hgc-I) and beta proteobacterial (beta-II) sequence clusters of freshwater bacteria that were also present in the lake water 16S rRNA gene library. At least two novel, deeply rooting alpha proteobacterial lineages were recovered from S. lacustris that showed <89% sequence similarity to known phylogenetic groups. Electron microscopical observations revealed that digested bacterial remnants were contained within food vacuoles of sponge archaeocytes, whereas the extracellular matrix was virtually free of bacteria. This study is the first molecular diversity study of a freshwater sponge and adds to a growing database on the diversity and community composition of sponge-associated microbial consortia. PMID- 16211325 TI - Spatial dominance and inorganic carbon assimilation by conspicuous autotrophic biofilms in a physical and chemical gradient of a cold sulfurous spring: the role of differential ecological strategies. AB - The community composition and ecophysiological features of microbial autotrophic biofilms were studied in Fuente Podrida, a cold sulfur spring located in East Spain. We demonstrated how different ecophysiological strategies, such as resistance and/or utilization of sulfide and oxygen, light adaptation, or resistance to high water flow, allow each of the microorganisms described to efficiently colonize several areas within the environmental gradient. In the zone of the spring constantly influenced by sulfide-rich waters, biofilms were formed by purple bacteria, cyanobacteria, and filamentous colorless sulfur bacteria. Purple bacteria showed higher photosynthetic efficiency per pigment unit than cyanobacteria, although they were dominant only in anoxic areas. Two filamentous cyanobacteria, strain UVFP1 and strain UVFP2, were also abundant in the sulfide rich area. Whereas the cyanobacterial strain UVFP2 shows a strategy based on the resistance to sulfide of oxygenic photosynthesis, strain UVFP1, additionally, has the capacity for sulfide-driven anoxygenic photosynthesis. Molecular phylogenetic analyses cluster the benthic strain UVFP1 with genus Planktothrix, but with no particular species, whereas UVFP2 does not closely cluster with any known cyanobacterial species. The colorless sulfur bacterium Thiothrix sp. extended throughout the zone in which both sulfide and oxygen were present, exhibiting its capacity for chemolithoautotrophic dark carbon fixation. Downstream from the source, where springwater mixes with well-oxygenated stream water and sulfide disappears, autotrophic biofilms were dominated by diatoms showing higher photosynthetic rates than cyanobacteria and, by a lesser extent, by a sulfide sensitive cyanobacterium (strain UVFP3) well adapted to low light availability, although in the areas of higher water velocity far from the river shore, the dominance shifted to crust-forming cyanobacteria. Both types of microorganisms were highly sensitive to sulfide impeding them from occupying sulfide-rich areas of the spring. Sulfide, oxygen, light availability, and water velocity appear as the main factors structuring the autotrophic community of Fuente Podrida spring. PMID- 16211326 TI - Colonization of tomato root seedling by Pseudomonas fluorescens 92 rkG5: spatio temporal dynamics, localization, organization, viability, and culturability. AB - The localization, viability, and culturability of Pseudomonas fluorescens 92 rkG5 were analyzed on three morphological root zones (root tip + elongation, root hair, and collar) of 3-, 5-, and 7-day-old tomato plants. Qualitative information about the localization and viability was collected by confocal laser scanning microscopy. Quantitative data concerning the distribution, viability, and culturability were obtained through combined dilution plating and flow cytometry. Colonization by P. fluorescens affected root development in a complex way, causing a general increase in the length of the collar and early stimulation of the primary root growth (3rd day), followed by a reduction in length (7th day). The three root zones showed different distribution, organization, and viability of the bacterial cells, but the distribution pattern within each zone did not change with time. Root tips were always devoid of bacteria, whereas with increasing distance from the apex, microcolonies or strings of cells became more and more prominent. Viability was high in the elongation zone, but it declined in the older parts of the roots. The so-called viable but not culturable cells were observed on the root, and their proportion in the distal (root tip + elongation) zone dramatically increased with time. These results suggest the existence of a specific temporal and spatial pattern of root colonization, related to cell viability and culturability, expressed by the plant-beneficial strain P. fluorescens 92 rkG5. PMID- 16211327 TI - Inoculation with the plant-growth-promoting rhizobacterium Azospirillum brasilense causes little disturbance in the rhizosphere and rhizoplane of maize (Zea mays). AB - Inoculation with Azospirillum brasilense exerts beneficial effects on plant growth and crop yields. In this study, a comparative analysis of maize (Zea mays) root inoculated or not inoculated with A. brasilense strains was performed in two soils. Colonization dynamics of the rhizobacteria were tracked in various root compartments using 16S rRNA-targeted probes and 4',6'diamidino-2-phenylindole staining, and the structure of bacterial populations in the same samples was analyzed by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction products of the 16S rRNA gene. Based on whole cell hybridization, a large fraction of the bacterial community was found to be active in both the rhizoplane-endorhizosphere and rhizosphere soil compartments, in both soil types. A DGGE fingerprint analysis revealed that plant inoculation with A. brasilense had no effect on the structural composition of the bacterial communities, which were also found to be very similar at the root tip and at zones of root branching. However, rhizobacterial populations were strongly influenced by plant age, and their complexity decreased in the rhizoplane-endorhizosphere in comparison to rhizosphere soil. A clone library generated from rhizosphere DNA revealed a highly diverse community of soil and rhizosphere bacteria, including an indigenous Azospirillum-like organism. A large proportion of these clones was only distantly related to known species. PMID- 16211328 TI - Nutrient movement and mycelial reorganization in established systems of Phanerochaete velutina, following arrival of colonized wood resources. AB - The effect of arrival of wood resources, pre-colonized by Coriolus versicolor, Phlebia radiata, Stereum hirsutum, and Vuilleminia comedens, on mycelial systems of Phanerochaete velutina was studied in trays of nonsterile soil in the laboratory over 5 months. Morphological responses were quantified non destructively using image analysis. In a parallel series of experiments, nutrient movement was also quantified non-destructively using (32)P monitoring with a scintillation probe and subsequently by destructively harvesting after 155 days. The presence of a fungus occupying a newly arriving resource had major effects on deployment of biomass and on the uptake and allocation of phosphorus in the established Pha. velutina system. The effects varied depending on the species occupying the new resource. Hyphal coverage was greater in the half of the system to which new resources were added. Following addition of new resources, there was massive redeployment of biomass away from regions with no new resource when the new resource was (1) uncolonized, (2) colonized by V. comedens, or (3) colonized by S. hirsutum (although to a lesser extent with the latter), but not with others. (32)P was taken up by Pha. velutina both in the vicinity of the inoculum and the new resource and was translocated to the new resource from both sites of uptake; however, the local supply contributed most. Bidirectional translocation also occurred. The results are discussed in relation to mycelial foraging strategies, nutrient translocation, and partitioning within mycelial cord systems. PMID- 16211329 TI - Identification and full sequence of an isolate of Alternanthera mosaic potexvirus infecting Phlox stolonifera. AB - A potexvirus was isolated from creeping phlox (Phlox stolonifera) plants from a commercial nursery in Pennsylvania. The virus was serologically related to clover yellow mosaic virus, plantain virus X, potato virus X, and potato aucuba mosaic virus, and was most closely related to papaya mosaic virus (PapMV). The sequence of a PCR fragment obtained with potexvirus group-specific primers was distinct from that of PapMV; the coat protein (CP) gene and 3' untranslated region (UTR) were closely related to Alternanthera mosaic virus (AltMV), previously reported only from Australia. The host range was similar to that of the Australian isolate (AltMV-Au), and the phlox isolate reacted strongly with antiserum to AltMV-Au. The full sequence of the phlox isolate was more closely related to PapMV throughout the genome than to any potexvirus other than AltMV-Au, for which only the CP and 3'UTR sequences are available. The phlox isolate was therefore named AltMV-PA (for Pennsylvania), and the full 6607 nt sequence is presented(1). Additional AltMV isolates from creeping phlox (AltMV-BR and AltMV-SP) and trailing portulaca (Portulaca grandiflora; AltMV-Po) were also isolated, suggesting that AltMV may be widespread, and may have been mis-diagnosed in the past as PapMV. AltMV has the potential to spread to other ornamental crops. PMID- 16211330 TI - Preliminary x-ray characterization of authentic providence virus and attempts to express its coat protein gene in recombinant baculovirus. AB - Providence Virus (PrV) is a non-envoloped, T = 4 icosahedral beta-tetravirus that undergoes autocatalytic cleavage of its coat protein precursor after capsid assembly. This is also a well characterized function of Nudaurelia capensis omega virus (NomegaV), a member of the related omegatetraviruses, whose x-ray structure has been determined. Virus-like particle (VLP) production of PrV in a recombinant baculovirus expression system was attempted to obtain high VLP yields for comparison of structural and autocatalytic active site properties between these virus groups. This resulted in insoluble aggregates of PrV coat protein even though NomegaV VLPs have been successfully produced in the same system. Betatetraviruses may be more dependent on compartmentalization and availability of their full-length genome for proper folding and assembly. However, crystals were grown of limited quantities of authentic PrV produced in cell culture and a partial X-ray data set collected to 3.8 A resolution. The virus particle position and orientation in the unit cell was determined by space group consideration and rotation function analysis. A phasing model, based on NomegaV, was developed to initiate the structure solution of PrV. PMID- 16211331 TI - Emergence of immune escape variant of mammary tumors that has distinct proteomic profile and a reduced ability to induce "danger signals". AB - Breast tumors are shaped, in part, by a process termed immunoediting which selects for immunologically evasive phenotypes. In the present study we used the rat neu-transgenic mouse model of breast cancer and its congenic non-transgenic parental strain, FVB, to explore the phenotype of tumors that emerge in the presence of an immune response directed against the neu antigen. When inoculated into parental FVB mice, a neu-overexpressing mouse mammary carcinoma (MMC) cell line isolated from spontaneous breast tumors of the FVB neu (FVBN202) transgenic mouse, elicited a neu-specific immune response resulting in a tumor rejection because of the presence of the rat neu antigen. However, a neu negative variant (ANV) of MMC arose after a long latency in spite of the neu-specific immune response. We show that compared to MMC, ANV tumor cells have a significantly reduced ability to secrete pro-inflammatory cytokines and the CCL5 chemokine, to express immunostimulatory chaperones, and they have a distinct expression of proteins involved in cell motility, and metabolic and signal transduction pathways. These studies suggest that tumor escape through immunoediting can not be explained by the loss of a single tumor antigen, but rather by a selection process of a tumor variant that has a reduced ability to induce "danger signals" together with up-regulation of proteins involved in the tumor survival. Based on these findings, we propose to target novel antigens over-expressed in the escape variant of breast tumors to treat primary tumor and to prevent tumor relapse. PMID- 16211333 TI - Frustration between syn- and anticlinicity in mixtures of chiral and non-chiral tilted smectic-C-type liquid crystals. AB - We study the effects of mixing ferroelectric and antiferroelectric liquid-crystal compounds (FLCs and AFLCs) when the former are strictly synclinic and the latter strictly anticlinic, i.e. one mixture component exhibits only SmC* and the other only SmC(a)* as tilted phase. Three different paths between syn- and anticlinicity were detected: transition directly between SmC* and SmC(a)*, transition via the SmC(beta)* and SmC(gamma)* subphases, or by "escaping" the clinicity frustration by reducing the tilt to zero, i.e. the SmA* phase is extended downwards in temperature, separating SmC* from SmC(a)* in the phase diagram. The most common path is the one via the subphases, demonstrating that these phases appear as a result of frustration between syn- and anticlinic and, consequently, between syn- and antipolar order. For assessing the role of chirality, we also replaced the FLC with non-chiral synclinics. With one of the AFLCs, the route via supbhases was detected even in this case, suggesting that chirality--although necessary--does not have quite the importance that has previously been attributed to the appearance of the subphases. The path chosen in the mixture study seemed to be determined mainly by the synclinic component, the subphase induction occurring only when the SmA*-SmC* transition was second order. PMID- 16211332 TI - Language, social, and executive functions in high functioning autism: a continuum of performance. AB - This study examined language and executive functions (EF) in high-functioning school-aged individuals with autism and individually matched controls. Relationships between executive, language, and social functioning were also examined. Participants with autism exhibited difficulty on measures of expressive grammar, figurative language, planning, and spatial working memory. A mixed profile of impaired and enhanced abilities was noted in set-shifting. While controls showed the typical increase in errors when shifting sets from an intra dimensional to an extra-dimensional stimulus, this pattern was not noted in participants with autism. Relationships between EF, language, and social performance were weak to non-existent. Implications for theories of core deficit in autism and dissociable nature of the language and executive impairments in autism are discussed. PMID- 16211334 TI - Nonlinear competition between asters and stripes in filament-motor systems. AB - A model for polar filaments interacting via molecular motor complexes is investigated which exhibits bifurcations to spatial patterns. It is shown that the homogeneous distribution of filaments, such as actin or microtubules, may become either unstable with respect to an orientational instability of a finite wave number or with respect to modulations of the filament density, where long wavelength modes are amplified as well. Above threshold nonlinear interactions select either stripe patterns or periodic asters. The existence and stability ranges of each pattern close to threshold are predicted in terms of a weakly nonlinear perturbation analysis, which is confirmed by numerical simulations of the basic model equations. The two relevant parameters determining the bifurcation scenario of the model can be related to the concentrations of the active molecular motors and of the filaments, respectively, which both could be easily regulated by the cell. PMID- 16211335 TI - Qualitative discrepancy between different measures of dynamics in thin polymer films. AB - We have used ellipsometry to measure the initial stages of interface healing in bilayer polystyrene films. We also used ellipsometry to measure the glass transition temperature T(g) of the same or identically prepared samples. The results indicate that as the film thickness is decreased, the time constant for the interface healing process increases, while at the same time the measured glass transition temperature in the same samples decreases as the film thickness is decreased. This qualitative difference in the behavior indicates that it is not always possible to make inferences about one probe of polymer dynamics from measurements of another. We propose a reason for this discrepancy based on a previously discussed origin for reduction in the T(g) value of thin films. PMID- 16211336 TI - Successful retreatment with infliximab in patients with prior severe infusion reactions. AB - Infusion of the antitumor necrosis factor-alpha chimeric monoclonal antibody infliximab can be associated with the development of severe infusion reactions (IR) during retreatment. We present the case of two rheumatoid arthritis patients with a history of severe acute IR to infliximab who subsequently underwent successful infusion using a prophylactic treatment with a combination of H1 and H2 receptor blockers, hydrocortisone, and diphenhydramine. PMID- 16211337 TI - Systemic sclerosis sine scleroderma presenting as pulmonary intersticial fibrosis. PMID- 16211338 TI - Rheumatoid lung nodulosis and osteopathy associated with leflunomide therapy. AB - BACKGROUND: Leflunomide (LEF) is indicated in adults for the treatment of active rheumatoid arthritis (RA). LEF inhibits dehydroorotate dehydrogenase, a key enzyme of the pyrimidine synthesis in activated lymphocytes. Among rare adverse effects, fatal interstitial lung disease has been recently reported during treatment of RA with LEF in Japan. Clinical trials outside Japan do not suggest that LEF causes an excess of pulmonary adverse effects. Development and increase of peripheral rheumatoid nodules in typical sites of RA patients following LEF therapy has been recently reported. OBJECTIVES: Two cases with new and accelerated development of rheumatoid lung nodulosis during LEF therapy were described in this study. METHODS: LEF treatment was administered to two male patients (77 and 66 years old) with long-standing active seropositive nodular RA with failure of multiple second line drugs and without lung involvement. Clinical and laboratory assessment using the American College of Rheumatology response criteria, chest computed tomography (CT), quantification of serum rheumatoid factor (RF), and monocyte count of peripheral blood along with routine laboratory follow up were performed on both patients before and during therapy. In case 1, a bone scan was performed due to sustained limbs pain. Open lung biopsy was performed in case 1 and core lung biopsy in case 2. RESULTS: Both patients achieved full clinical remission during 2 months of LEF therapy. In case 1, the first complaints were limbs pain after 10 months of treatment associated with intensive bone uptake on a bone scan consistent with hypertrophic pulmonary osteopathy. Productive cough developed after 3 months of the therapy in case 2. Initially, these complaints were not attributed to therapy. New lung disease was present on CT with cherry-like progressive cavitary nodules, predominantly involving the basal segments of the right lung. The first lung lesions were found by CT 13 months (case 1) and 7 months (case 2) after the beginning of therapy and were erroneously related to bronchiectasia in case 2. In both cases, the lung biopsy showed necrosis surrounded by epithelioid mononuclear inflammation with giant cells, consistent with rheumatoid lung node. The time that elapsed between the beginning of the first symptoms to LEF discontinuation was very long: 13 months in case 1 and 24 months in case 2. Discontinuation of LEF therapy was followed by an arrest in growth of lung nodules, resolution of limb pain, and gradual improvement of bone scan. A significant decrease of monocyte count and RF level in peripheral blood was observed during LEF therapy in both cases. CONCLUSION: For the first time, we described rheumatoid lung nodulosis as complication of successful LEF therapy for RA. Hypertrophic pulmonary osteopathy with severe limbs pain and dry cough were the first manifestations of the lung nodulosis. Monocytopenia during LEF therapy is proposed to be involved in pathogenesis of this rare complication of LEF therapy. PMID- 16211339 TI - Topical lidocaine helps reduce pain of digital ulcers in systemic sclerosis (scleroderma). PMID- 16211340 TI - Frequent insectivory by two guenons (Cercopithecus lhoesti and Cercopithecus mitis) in the Kalinzu Forest, Uganda. AB - I investigated the feeding ecology of l'Hoest's monkeys (Cercopithecus lhoesti) and blue monkeys (Cercopithecus mitis) in the Kalinzu Forest, Uganda. Although forest guenons are generally thought to be frugivores or folivores, these two guenons spent a large proportion of their time feeding on invertebrates. The l'Hoest's monkey and the blue monkey spent as much as 66 and 50% of their time on insectivory, respectively. These proportions of time spent on invertebrate feeding are higher than those reported elsewhere for forest guenons. L'Hoest's monkeys mainly utilized the area near the ground for invertebrate feeding, while blue monkeys utilized the area around 20 m above the ground. It seemed that guenons have an ability to change their diet according to the environment. PMID- 16211341 TI - Biliary complications after a right-lobe living donor liver transplantation. AB - Right-lobe living donor liver transplantation (RL-LDLT) has become an acceptable procedure for adult patients with end-stage liver disease in this decade. However, biliary complications in RL-LDLT remain a serious problem: the incidence of anastomotic biliary leakage and stricture after RL-LDLT is reported to be 4.7% 18.2% and 8.3%-31.7%, respectively. The incidence varies according to the type of biliary reconstructions between Roux-en-Y hepaticojejunostomy and duct-to-duct biliary reconstruction. The anatomical biliary diversity of a right-lobe graft makes it difficult to reconstruct the biliary system. Indeed, most biliary strictures in patients with duct-to-duct reconstruction develop in multibranched fashion. In this regard, endoscopic biliary stenting appears to be efficacious for treating multibranched biliary strictures because multiple stenting permits the drainage of each segmental branch of the stricture. In this review, we describe various aspects of biliary complications occurring in RL-LDLT and their treatment. PMID- 16211342 TI - A prospective study of the prevalence of gastroesophageal reflux disease and confounding factors. AB - BACKGROUND: The prevalence of gastroesophageal reflux disease (GERD) has reportedly risen in recent years. Difficulties associated with endoscopic diagnosis mean it is not easy to determine its precise prevalence. A prospective study of the prevalence of endoscopy-positive GERD (EP-GERD) was conducted at Higuchi Hospital, a general hospital in Northwestern Kyushu, Japan. The study also correlated factors that might affect prevalence (age, sex, and functions of the gastroesophageal junction). METHODS: From consecutive patients undergoing endoscopic examination at Higuchi Hospital between January 2000 and April 2003, 1234 patients without severe complications were examined for the possible presence of GERD. Patients were stratified by age and sex, and the prevalence in each group ascertained. EP-GERD was defined on the four-level scale of the Los Angeles classification. Endoscopic classification of gastroesophageal flap valve ([GEFV] functional anomalie; using a four-level scale), was done as proposed by Hill et al. in 1996, to assess flap-valve morphology. Six items were evaluated: (1) symptoms and primary diseases; (2) prevalence of development of EP-GERD classified by age and sex; (3) endoscopic morphology of the GEFV as an expression of the functions of the gastroesophageal junction, and its prevalence by age and sex; (4) regression analysis and Spearman's rank correlation of GEFV and EP-GERD grades; (5) prevalence of EP-GERD and GEFV stratified by age and analyzed; and (6) multiple regression analysis of EP-GERD and explanatory variables (age, sex, and GEFV). RESULTS: The overall prevalence of EP-GERD was 5.8% (72/1234) and this patient group was dominated by men. Aging had minimal effect on prevalence in men, but the prevalence rose among women as they aged. The age-stratified prevalence of GEFV functional anomalies was similar to the age-stratified prevalence of EP-GERD in both sexes. The correlation between EP-GERD and GEFV functional anomalies was high regardless of sex. CONCLUSIONS: We postulate that the mechanisms leading to the development of GEFV functional anomalies in men are different from those in women. Future evaluations of EP-GERD should also observe GEFV function. PMID- 16211343 TI - Association between interleukin-1beta-511C/T polymorphism and reflux esophagitis in Japan. AB - BACKGROUND: Interleukin-1beta (IL-1beta) gene polymorphisms are related to hypochlorhydria and increase the risk of gastric cancer in the presence of Helicobacter pylori infection. However, little information is available about the genetic risk factors of reflux esophagitis. In this study we investigated its association with the IL-1beta polymorphisms. METHODS: We examined 48 patients with reflux esophagitis and 96 control subjects, 89 with gastric cancer. IL-1beta 511C/T genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: The frequency of IL 1beta-511T alleles was significantly higher in reflux esophagitis patients (57.3%) than in controls (41.1%) (P = 0.0215, chi(2) = 5.289). The frequency of IL-1beta-511T/T genotypes was also significantly higher in reflux esophagitis patients (31.3%) than in controls (15.6%). The odds ratio and the 95% confidence interval were 4.000 and 1.393-11.486, respectively. The frequency of IL-1beta 511T/T genotypes was significantly higher in reflux esophagitis patients (31.3%) than in gastric cancer patients (21.4%). The odds ratio and the 95% confidence interval were 2.961 and 1.054-8.316, respectively. CONCLUSIONS: IL-1beta-511T was associated with reflux esophagitis having hyperacidity. Differences of genetic background regarding gastric acid secretion may exist between Japanese and Caucasians. PMID- 16211344 TI - Alteration of intestinal epithelial function by intraepithelial lymphocyte homing. AB - BACKGROUND: Intimate cross-talk may take place between intestinal epithelial cells and intraepithelial lymphocytes (IEL). The purpose of this study was to analyze the influence of lymphocyte migration into the epithelium on epithelial function, using an in vitro "IEL homing" model. METHODS: Molecular expression on epithelial cells was analyzed by flow cytometry. The barrier function of the epithelial monolayer was assessed by transepithelial electrical resistance. Cytokine production was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: (1) IEL homing into the epithelia induced significant phenotypic changes in epithelial cells; upregulation of MHC class I, and II, intercellular adhesion molecule (ICAM)-1, and CD44. IEL-derived interferon-gamma (IFN-gamma) could partially account for this alteration, as a neutralizing antibody (Ab) against IFN-gamma inhibited the upregulation of these molecules, except for CD44. (2) A marked fall in transepithelial electrical resistance was observed 4 h after IEL homing started, and Ab against IFN-gamma slightly inhibited this fall in resistance. (3) The production of interleukin (IL)-8 and IFN-gamma inducible protein-10 (IP-10), but not transforming growth factor (TGF)-beta1 or tumor necrosis factor (TNF)-alpha, in the epithelial monolayer was markedly induced after IEL homing in a basolaterally polarized fashion. IEL-conditioned media also induced the production of these cytokines in epithelial cells, thus suggesting that IEL-derived soluble factor(s) induce epithelial chemokine production. CONCLUSIONS: Under inflammatory conditions, IEL obviously interact with epithelial cells and upregulate adhesion molecules, alter barrier function, and enhance chemokine production. Because such alterations may increase epithelial permeability to luminal antigens or accelerate the migration of other inflammatory cells, our results suggest that IEL have a critical role in mucosal immunity. PMID- 16211346 TI - Erectile dysfunction in patients with chronic viral liver disease: its relevance to protein malnutrition. AB - BACKGROUND: In patients with chronic liver disease (CLD), quality of life is generally accepted as poor, especially for physical function. However, sufficient data regarding erectile function has not been shown in patients with CLD. The international index of erectile function (IIEF) is widely used to assess erectile function, and a short form of the IIEF was recently developed (IIEF-5). Using this questionnaire, we evaluated erectile dysfunction (ED) in patients with CLD. METHODS: A total of 117 Japanese patients (64 with chronic hepatitis [CH] and 53 with liver cirrhosis [LC]) were analyzed. The etiologies were hepatitis B virus (HBV) in 21, HCV in 94, and non-B non-C in 2. The IIEF-5 and Medical Outcomes Study Short Form 36 (SF-36) were administered to the patients, and biochemical analyses for items serum albumin, prothrombin time, bilirubin, and ammonia were also performed. RESULTS: The incidence of ED was 85% in the total cohort with CLD, 78% in those with CH, and 92% in those with LC (P < 0.05 between CH and LC). ED was found in 50% of CLD patients under age 50 years, in 79% aged 50-59, and in 100% aged over 60 (P, overall <0.001). The scores for ED severity correlated with increasing grades of a modified Child-Pugh classification (P < 0.05). Simple regression analysis showed age (P < 0.01), physical function (P < 0.001), role physical (P < 0.001), and social functioning (P < 0.05) on the SF-36, and serum albumin (P < 0.001) as significant determinants of ED. Multiple regression analysis identified age (P < 0.001) and serum albumin (P < 0.001) as independent significant factors that determined ED. CONCLUSIONS: These data clearly demonstrate that liver disease is the cause of ED in patients with CLD, and serum protein status could be relevant to this condition in these patients. PMID- 16211345 TI - Helicobacter pylori infection increases the risk of colorectal adenoma and adenocarcinoma, especially in women. AB - BACKGROUND: Recent reports suggest that Helicobacter pylori infection can potentially increase the risk of colorectal cancer. The purpose of this study was to assess the association between H. pylori infection and the risk of colorectal adenoma and adenocarcinoma, and to evaluate any differences on the basis of sex. METHODS: The subjects were 669 (40- to 80-year-old) patients who underwent both barium enema examination and total colonoscopy, and who were evaluated for H. pylori infection by (13)C-urea breath test, urease test, or histological diagnosis of biopsied gastric specimens. There were 142 H. pylori-negative and 527-positive patients. The odds ratios (ORs) for H. pylori-positive patients with colorectal adenoma and adenocarcinoma, and for tumor patients with either adenoma or adenocarcinoma were calculated. RESULTS: Among the H. pylori-negative patients, there were 52 patients without tumor, 63 with adenoma, 27 with adenocarcinoma, and 90 with tumor. Among the H. pylori-positive patients, there were 136, 264, 127, and 391 patients respectively. Pooling all subjects, those infected with H. pylori had a significantly increased OR for adenoma, adenocarcinoma, or tumor, compared to H. pylori-free patients (OR, 1.60, 1.80, and 1.66, respectively). For female H. pylori-positive subjects, the risk of having adenocarcinoma or tumor was significantly higher than that for their H. pylori-free counterparts, while for male H. pylori-positive and -negative subjects, there was no such significant difference. CONCLUSIONS: The results therefore suggest that, in patients aged 40-80 years, H. pylori infection increased the risk of colorectal adenoma and adenocarcinoma, with significantly higher risks for female patients. PMID- 16211347 TI - Determinants of serum ALT normalization after phlebotomy in patients with chronic hepatitis C infection. AB - BACKGROUND: Phlebotomy is performed to reduce excessive iron accumulation in hepatic tissue. We studied serum alanine aminotransferase (ALT) normalization rates and 50% reduction in initial serum ALT (ALT(50%) reduction rate) in patients with hepatitis C viral (HCV) infection and investigated the factors that influenced the response to phlebotomy therapy. METHODS: We evaluated 23 consecutive patients with HCV infection who underwent phlebotomy. Phlebotomy was performed a few times per week, then a few times per month, and 200-400 ml of blood was removed at each session, depending on the clinical response. During the course of therapy, hemoglobin (Hb), serum ALT, and ferritin levels were assessed monthly. RESULTS: In patients with Hb of less than 11 g/dl, the ALT(50%) reduction rate was 87.5%. In patients with a serum ferritin level of less than 10 g/dl the ALT(50%) reduction rate was 83.3%. In patients with Hb of less than 11 g/dl, the ALT normalization rate was 50%, and in those with a serum ferritin level of less than 10 g/dl, the ALT normalization rate was 41.7%. Multivariate analysis identified ALT less than 100 IU/l at the start of phlebotomy as an independent factor associated with ALT normalization. Of the 7 patients who showed no response to phlebotomy, 85.7% were obese (body mass index > or =25 kg/m(2)), and 40% showed more than 30% steatosis on liver histology. The cumulative ALT normalization rate in relation to the total volume of blood loss was 43.9% with a blood less or more than 3 l, and thus was optimal above 3 l. CONCLUSIONS: Although the sample number was relatively small, the results of our study suggest that phlebotomy is effective therapy for HCV patients who are nonobese, show little or no steatosis on liver histology, and have a baseline serum ALT level of less than 100 IU/l. PMID- 16211350 TI - Re: Possible prevention of fulminant hepatic failure in four children with acute severe hepatitis. PMID- 16211348 TI - Usefulness of endoscopic ultrasound-guided fine-needle aspiration biopsy for the diagnosis of pancreatic cancer. AB - BACKGROUND: Endoscopic ultrasonography-guided fine-needle aspiration biopsy (EUS FNAB) has come into widespread use, mainly in Western countries, as an efficient and safe method for the cytologic or histologic diagnosis of pancreatic cancer. However, it still has received relatively little attention in Japan. To evaluate the clinical status of EUS-FNAB in Japan, we retrospectively analyzed the results with regard to the ability of EUS-FNAB to diagnose pancreatic cancer, as well as its safety. METHODS: A total of 52 patients (37 male, 15 female; mean age, 62.5 years; range, 33-85 years) with focal pancreatic lesions underwent EUS-FNAB at our group of hospitals in one region of Japan. Final diagnosis was confirmed by histologic examination of surgical specimens or clinical follow-up. RESULTS: The final diagnoses were malignant tumors in 32 patients and benign ones in 20. Insertion of the needle into the lesion was successful in 50 of the 52 patients (96.2%). Adequate specimens were obtained by EUS-FNAB from 47 of the 50 pancreatic lesions (94.0%). With five false-negative and no false-positive results, the accuracy, sensitivity, specificity, and positive and negative predictive values were 89.4%, 82.1%, 100%, 100%, and 79.2%, respectively. No complications occurred. CONCLUSIONS: EUS-FNAB is an efficient and safe method for the histologic diagnosis of pancreatic cancer. It should be considered as one of the indispensable modalities for the histological diagnosis of pancreatic cancer in Japan, as it is in Western countries. PMID- 16211351 TI - Prevalence of noncardiac chest pain in Japanese patients with recurrent chest pain. PMID- 16211352 TI - Crohn's disease in Turner's syndrome with X-chromosomal mosaicism of 45 XO and 47 XXX. PMID- 16211353 TI - Angiomyolipoma of the liver presenting a homogeneously hypoechoic mass. PMID- 16211354 TI - No mutations in the tyrosine kinases of human hepatic, pancreatic, and gastric cancer cell lines. PMID- 16211355 TI - Helicobacter pylori and colorectal neoplasm: a mysterious link? PMID- 16211356 TI - Complications of endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS FNAB) for pancreatic lesions. PMID- 16211357 TI - Lymphangiogenesis in human gynaecological cancers. AB - The metastatic spread of tumor cells is responsible for the majority of cancer deaths, and with few exceptions, all cancers can metastasize. Clinical findings have for a long time suggested that by providing a pathway for tumor cell dissemination, tumor-associated lymphatics act as key components of metastatic spread. This is believed to occur principally via pre-existing and possibly also newly formed lymphatics (lymphangiogenesis). Increased expression of vascular endothelial growth factor-C (VEGF-C) and VEGF-D in primary tumors correlates with increased dissemination of tumor cells to regional lymph nodes (LNs) in a variety of human carcinomas. Here we will review the mechanisms of lymphangiogenesis, particularly in the context of metastatic tumor spread, and will critically examine the role of VEGF-C and VEGF-D in this process in gynaecological cancers. Potential anti-lymphangiogenic strategies are also discussed. PMID- 16211358 TI - Vascular biology in implantation and placentation. AB - Pregnancy leads to dramatic changes of the vascular system of the mother and enables the development of a completely new vascular system within the growing embryo including the formation of the placenta as the exchange organ between both circulations. Besides a general adaptation of the maternal blood system, the uterine spiral arteries display the greatest changes. Within placental villi angiogenesis as well as vasculogenesis can be found already a few weeks after implantation. Both systems in parallel will determine the blood flow within the placental villi and the intervillous space. Finally, compromised blood flow on either side of the placental membrane will not only lead to fetal malnutrition, but will also trigger morphological changes of the villous trees. This review tries to cover all the above-mentioned topics and will try to depict the consequences of poor placentation on mother and fetus. PMID- 16211359 TI - Recent advances in endometrial angiogenesis research. AB - This review summarises recent research into the mechanisms and regulation of endometrial angiogenesis. Understanding of when and by what mechanisms angiogenesis occurs during the menstrual cycle is limited, as is knowledge of how it is regulated. Significant endometrial endothelial cell proliferation occurs at all stages of the menstrual cycle in humans, unlike most animal models where a more precise spatial relationship exists between endothelial cell proliferation and circulating levels of oestrogen and progesterone. Recent stereological data has identified vessel elongation as a major endometrial angiogenic mechanism in the mid-late proliferative phase of the cycle. In contrast, the mechanisms that contribute to post-menstrual repair and secretory phase remodelling have not yet been determined. Both oestrogen and progesterone/progestins appear to have paradoxical actions, with recent studies showing that under different circumstances both can promote as well as inhibit endometrial angiogenesis. The relative contribution of direct versus indirect effects of these hormones on the vasculature may help to explain their pro- or anti-angiogenic activities. Recent work has also identified the hormone relaxin as a player in the regulation of endometrial angiogenesis. While vascular endothelial growth factor (VEGF) is fundamental to endometrial angiogenesis, details of how and when different endometrial cell types produce VEGF, and how production and activity is controlled by oestrogen and progesterone, remains to be elucidated. Evidence is emerging that the different splice variants of VEGF play a major role in regulating endometrial angiogenesis at a local level. Intravascular neutrophils containing VEGF have been identified as having a role in stimulating endometrial angiogenesis, although other currently unidentified mechanisms must also exist. Future studies to clarify how endometrial angiogenesis is regulated in the human, as well as in relevant animal models, will be important for a better understanding of diseases such as breakthrough bleeding, menorrhagia, endometriosis and endometrial cancer. PMID- 16211360 TI - Vascular development in endometriosis. AB - Endometriosis, defined as the presence of endometrial tissue outside the uterus, is an estrogen-dependent disease which causes pelvic pain and subfertility in women of reproductive age. The condition has a dramatic impact on the professional, social and marital life of sufferers. Direct and indirect evidence suggests that angiogenesis is required for the development and persistence of endometriosis. In this review the state-of-the-art with regard to our understanding of the role of angiogenesis in the ectopic implantation and survival of menstrual endometrial tissue will be discussed. PMID- 16211361 TI - Progestogen only contraception and endometrial break through bleeding. AB - Progestogen only contraceptives (POC) provide a safe and effective method of fertility regulation. Unfortunately, they are commonly associated with the problem of endometrial break through bleeding (BTB), often leading to discontinuation of use. An increase in endometrial vascular fragility has been demonstrated as an important mechanism that contributes to BTB but our understanding of the interaction between exogenous steroid use and endometrial vasculature remains incomplete. This review sets out to describe a number of commonly used POC, their effects on endometrial morphology and possible molecular and cellular mechanisms that may lead to unscheduled bleeding. PMID- 16211362 TI - Sex steroid regulation of angiogenesis in breast tissue. AB - Angiogenesis is essential for normal function in the female reproductive tract and a prerequisite for growth and metastasis of solid tumors. Several factors, both inducers and inhibitors, play essential roles in the regulation of the angiogenic process. Exposure to sex steroids increases the risk of breast cancer but the mechanisms are poorly understood and the importance of angiogenesis in breast carcinogenesis is undefined. In the female reproductive tract ovarian hormones tightly regulate angiogenesis. The breast is also a target organ for sex steroids but very little is known about sex steroid effects on angiogenesis in normal breast tissue and breast cancer. In this review several regulators of angiogenesis, and their relation to sex steroids, in breast tissue are discussed. Increased knowledge in this area is of utmost importance for future therapeutic treatment options and for breast cancer prevention. PMID- 16211363 TI - Angiogenesis in normal and neoplastic ovaries. AB - Ovarian physiology is intricately connected to hormonally regulated angiogenic response. Recent advances in the post genomic revolution have significantly impacted our understanding of ovarian function. In an angiogenesis perspective, the ovary offers a unique opportunity to unravel the molecular orchestration of blood vessel development and regression under normal conditions. A majority of ovarian cancers develop from the single layer of epithelium surrounding the ovaries. Angiogenesis is critical for the development of ovarian cancer and its peritoneal dissemination. The present review summarizes recent findings on the angiogenic response in neoplastic ovaries and discusses the prospects of using anti-angiogenic approaches to treat ovarian cancer. PMID- 16211365 TI - Phase I study of vinflunine administered as a 10-minute infusion on days 1 and 8 every 3 weeks. AB - Vinflunine is a novel vinca alkaloid developed through the selective modification of vinorelbine using super-acidic chemistry. In preclinical testing, vinflunine demonstrated significantly enhanced anti-tumour activity in human tumour xenograft models when compared to its parent compound. A phase I study was conducted to evaluate the safety and toxicity of vinflunine administered as a 10 minute intravenous infusion on days 1 and 8 every three weeks. Sixteen patients with advanced solid tumours were treated. Two of four patients experienced dose limiting toxicities (DLT) at 190 mg/m2 and this was established as the maximum tolerated dose (MTD). At the MTD, the DLT of vinflunine consisted of constipation and neutropenia. Fatigue was notable but not dose limiting. No objective responses were observed. A dose of 170 mg/m2 given on a day 1 and 8 schedule every three weeks would be suitable for future studies. PMID- 16211366 TI - Quantitative single-cell RT-PCR and Ca2+ imaging in brain slices. AB - We have established a quantitative reverse transcriptase-PCR (RT-PCR) approach for the analysis of RNA transcript levels in individual cells of living brain slices. Quantification is achieved by using rapid-cycle, real-time PCR protocols and high-resolution external cDNA standard curves for the gene of interest. The method consists of several procedures, including cell soma harvest, reverse transcription, and an optimized cDNA purification step, which allowed us to quantify transcripts in small types of neurons, like cerebellar granule cells. Thus, we detected in single granule cells an average of 20 transcript copies of the housekeeping gene glyceraldehyde-3-phosphate-dehydrogenase. We combined two photon calcium imaging and quantitative RT-PCR in single Purkinje and granule cells, respectively, and identified distinct glutamate receptor-dependent Ca2+ responses in these two cell types. The approach was further tested by profiling the expression of the ionotropic glutamate receptor subunits NR2B and NR2C in the cerebellum. Our study revealed a developmental switch from an average of 15 NR2B copies/cell at postnatal day 8 (P8) to about five NR2C copies/cell after P26. Taken together, our results demonstrate that the new method is rapid, highly sensitive, provides reliable results in neurons of various sizes, and can be used in combination with Ca2+ imaging. PMID- 16211367 TI - Phosphodiesterase inhibition promotes the transition from compensated hypertrophy to cardiac dilatation in rats. AB - The cellular signaling pathways responsible for the transition from compensated left ventricular hypertrophy (LVH) to LV dilatation (remodeling) and heart failure are unclear. As chronic administration of a beta-adrenoreceptor (beta-AR) agonist mediates the premature onset of cardiac remodeling without myocyte necrosis or myocardial dysfunction in LVH, we suggest that beta-AR activation is critical in promoting the transition from compensated LVH to cardiac dilatation. However, beta-AR mediated effects in the heart can occur via either the cyclic adenosine monophosphate (cAMP) system or via cAMP independent signaling pathways. To determine the role of cAMP in promoting adverse cardiac chamber remodeling, we evaluated whether phosphodiesterase inhibition (PDEI) promotes LV dilatation in rats with compensated LVH. The impact of chronic administration of the PDEI, pentoxifylline, on LV remodeling and function was assessed in spontaneously hypertensive rats (SHR) with compensated LVH. The PDEI mediated inotropic effects and increased cAMP concentrations in SHR. This dose of the PDEI administered for 4 months to SHR did not modify LV weight or influence intrinsic myocardial systolic function (as assessed in the absence of the PDEI) in SHR. However, the PDEI mediated the development of a right shift in LV end diastolic (LVED) pressure-internal dimension and LVED pressure-volume relations, LV wall thinning, and increments in myocardial soluble (non-cross-linked) collagen concentrations. In conclusion, chronic PDEI administration induces adverse geometric and interstitial cardiac remodeling in SHR, a finding that supports the notion that the beta-AR-cAMP system is important in mediating the progression to heart failure by promoting interstitial remodeling and LV dilatation in LVH. PMID- 16211369 TI - Isolated idiopathic ovarian vein thrombosis: a rare case. AB - This article describes the case of a 36-year-old woman who presented with a 2-day history of abdominal pain and nausea. Laboratory investigations revealed no abnormalities. Ovarian vein thrombosis was detected on ultrasound examination, and computed tomography of the pelvis and abdomen confirmed this finding. The patient was placed on anticoagulant therapy with unfractionated heparin and warfarin. Within 40 days her thrombosis improved. Laboratory testing for coagulation disorder revealed nothing abnormal. Ovarian vein thrombosis must be recognized and treated early to reduce the risk of serious complications. This case of idiopathic ovarian vein thrombosis underlines the importance of including this condition in the list of differential diagnoses for abdominal pain. PMID- 16211368 TI - Molecular properties of the SLC13 family of dicarboxylate and sulfate transporters. AB - The SLC13 gene family consists of five members in humans, with corresponding orthologs from different vertebrate species. All five genes code for sodium coupled transporters that are found on the plasma membrane. Two of the transporters, NaS1 and NaS2, carry substrates such as sulfate, selenate and thiosulfate. The other members of the family (NaDC1, NaDC3, and NaCT) are transporters for di- and tri-carboxylates including succinate, citrate and alpha ketoglutarate. The SLC13 transporters from vertebrates are electrogenic and they produce inward currents in the presence of sodium and substrate. Substrate independent leak currents have also been described. Structure-function studies have identified the carboxy terminal half of these proteins as the most important for determining function. Transmembrane helices 9 and 10 may form part of the substrate permeation pathway and participate in conformational changes during the transport cycle. This review also discusses new members of the SLC13 superfamily that exhibit both sodium-dependent and sodium-independent transport mechanisms. The Indy protein from Drosophila, involved in determining lifespan, and the plant vacuolar malate transporter are both sodium-independent dicarboxylate transporters, possibly acting as exchangers. The purpose of this review is to provide an update on new advances in this gene family, particularly on structure function studies and new members of the family. PMID- 16211370 TI - Concomitant resection of congenital vaginal septum during the tension-free vaginal tape procedure. AB - This case report illustrates how the tension-free vaginal tape (TVT) procedure can be successfully combined with another gynecological intervention. A 38-year old patient had been diagnosed with didelphys uterus during cesarean section. She came to us for treatment of confirmed genuine stress incontinence. In conjunction with the TVT procedure, we resected the congenital vaginal septum. The patient has remained continent for over 1 year. PMID- 16211371 TI - Four pregnancies and two deliveries after unilateral orchidectomy and chemotherapy for testicular embryonal carcinoma. AB - This case report describes a 33-year-old patient diagnosed with left-sided testicular embryonic carcinoma with vascular invasion. Unilateral orchiectomy was performed and the patient subsequently underwent chemotherapy. He retained fertility and later fathered healthy children. PMID- 16211372 TI - Dedication to professor Julian Davies. PMID- 16211373 TI - Associations of TV viewing and physical activity with the metabolic syndrome in Australian adults. AB - AIMS/HYPOTHESIS: We analysed a sample of Australian adults to determine the strength of associations of TV viewing and participation in physical activity with the metabolic syndrome. METHODS: This population-based cross-sectional study included 6,241 adults aged > or =35 years who were free from diagnosed diabetes mellitus and self-reported ischaemic disease and were not taking lipid-lowering or antihypertensive drugs. The metabolic syndrome was defined according to the 1999 World Health Organization criteria. Participants self-reported TV viewing time and physical activity time for the previous week. RESULTS: The adjusted odds ratio of having the metabolic syndrome was 2.07 (95% CI 1.49-2.88) in women and 1.48 (95% CI 0.95-2.31) in men who watched TV for >14 h per week compared with those who watched < or =7.0 h per week. Compared with those who were less active (<2.5 h per week), the odds ratio for the metabolic syndrome was 0.72 (95% CI 0.58-0.90) in men and 0.53 (95% CI 0.38-0.74) in women who were active (> or =2.5 h per week). Longer TV viewing (>14 h per week) was associated with an increased risk of insulin resistance, obesity and dyslipidaemia in both men and women. A total physical activity time of > or =2.5 h per week was associated with a reduced prevalence of both insulin resistance and dyslipidaemia in both sexes and reduced prevalence of both obesity and hypertension in women. CONCLUSIONS/INTERPRETATION: Increased TV viewing time was associated with an increased prevalence of the metabolic syndrome, while physical activity was associated with a reduced prevalence. Population strategies addressing the metabolic syndrome should focus on reducing sedentary behaviours such as TV viewing, as well as increasing physical activity. PMID- 16211374 TI - Proinsulin levels and the proinsulin:c-peptide ratio complement autoantibody measurement for predicting type 1 diabetes. AB - AIMS/HYPOTHESIS: We investigated whether random proinsulin levels and proinsulin:C-peptide ratio (PI:C) complement immune and genetic markers for identifying relatives at high risk of type 1 diabetes. MATERIALS AND METHODS: During an initial sampling, random glycaemia, proinsulin, PI:C and HLA DQ genotype were determined in 561 non-diabetic first-degree relatives who had been positive for islet autoantibodies on one or more occasions and in 561 age- and sex-matched persistently antibody-negative relatives. RESULTS: During follow-up (median 62 months), 46 relatives with antibodies at entry developed type 1 diabetes. At baseline, antibody-positive relatives (n=338) had higher PI:C values (p<0.001) than antibody-negative subjects with (n=223) or subjects without (n=561) later seroconversion. Proinsulin and PI:C were graded according to risk of diabetes as expressed by positivity for (multiple) antibodies or IA-2 antibodies, especially in persons carrying the high-risk HLA DQ2/DQ8 genotype and in prediabetic relatives. In the presence of multiple or IA-2 antibodies, a PI:C ratio exceeding percentile 66 of all antibody-negative relatives at entry (n=784) conferred a 5-year diabetes risk of 50% and 68%, respectively (p<0.001 vs 13% for same antibody status with PI:C75 years of age) and for those with high levels of serum lactate dehydrogenase (LDH) (more than two times the normal values). Patients with high-risk cytogenetic abnormalities according to the IPSS risk criteria showed better overall survival than those with intermediate-risk abnormalities. When analysed according to the IPSS risk classification, high-risk patients had worse survival prospects following decitabine therapy than those with intermediate risk; however, compared to the originally reported IPPS outcomes for high-risk patients, they probably showed better survival. During the treatment period, 18% of the patients progressed towards acute leukaemia. Decitabine showed a rather low toxicity profile in this elderly patient group. In conclusion, low-dose decitabine is an active drug for the treatment of MDS patients, even for those older than 75 years with bad prognostic characteristics. PMID- 16211385 TI - Degeneration in dysplastic hips. A computer tomography study. AB - BACKGROUND: Hip dysplasia is considered pre-osteoarthritic, causing degeneration in young individuals. OBJECTIVE: To determine the pattern of degenerative change in moderate to severely dysplastic hips in young patients. DESIGN AND PATIENTS: One hundred and ninety-three consecutively-referred younger patients with hip pain believed to be caused by hip dysplasia constituted the study cohort. The average age was 35.5 years (range, 15-61 years). They were examined by close-cut transverse pelvic and knee computed tomography and antero-posterior radiographs (CT). We identified 197 hips with moderate to severe dysplasia, and 78 hips with normal morphology in the study cohort, whilst 111 hip joints were borderline dysplastic according to preset definitions. Comparative analyses of anatomy and distribution of degeneration between dysplastic and normal hips in the study cohort were performed. RESULTS: In dysplastic hips the anterior acetabular sector angle was significantly and inversely associated to femoral anteversion (p < 0.001). The center-edge (CE) angle, the acetabular angle (AA), and the acetabular depth ratio (ADR) were significantly interrelated (p < 0.001; correlation coefficients ranging from -0.8 to 0.7). Fifty-one hips were subluxated (24R/27L). There were no cases of complete dislocation. The formation of subchondral cysts or osteophytes in dysplastic hips was significantly associated with reduced minimum joint space width (p ranging from 0.005 to 0.02). However, in 67 hips with acetabular cysts, only 6 hips had minimum joint space widths = 2.0 mm (8.9%) in the coronal plane. In 96 cases with acetabular cysts found in the sagittal plane, 43 cases had minimum joint space widths = 2.0 mm (44.7%). Bony rim detachment at the site of labral insertion was recorded in 30 hips. Twenty-three of these were dysplastic (p = 0.01). CONCLUSIONS: Degeneration was most often found in the anterolateral part of the dysplastic hip joints. Most cysts were located above the transition zone between the bony and the fibrocartilaginous acetabulum, and we found a significantly- increased number of cases with avulsed bony fragments at the antero-lateral labral insertion in dysplastic hips compared to normal hips. It seems likely that the early degenerative process in dysplastic hips originates at the watershed zone between the acetabular labrum and the acetabular cartilage in response to subluxation and femoroacetabular impingement. PMID- 16211387 TI - Expression of a novel somite-formation-related gene, AmphiSom, during amphioxus development. AB - A novel gene, AmphiSom, was identified in amphioxus Branchiostoma belcheri tsingtauense. Its sequence and developmental expression pattern were determined. AmphiSom transcripts were first detected in the presomitic mesoderm at the late gastrula stage and reached the highest level in the forming and nascent somites in neurulae. However, the expression of AmphiSom was rapidly down-regulated after somites were formed. It was maintained in the most anterior somite and most posterior somite at neurula stages. By 48 h, AmphiSom transcripts were detected only in the developing tail bud but were no longer detected in 72-h larva. Our data demonstrated that the AmphiSom gene is expressed during the development of somites in amphioxus and could play a role in somite formation. PMID- 16211388 TI - Evidence of a role for tyrosine dephosphorylation in the control of postgermination arrest of development by abscisic acid in Arabidopsis thaliana L. AB - In the present paper evidence is presented indicating that tyrosine dephosphorylation is a key regulatory mechanism in postgermination arrest of Arabidopsis thaliana L. seed development mediated by abscisic acid (ABA). By using phenylarsine oxide (PAO), an inhibitor of tyrosine phosphatases, the sensitivity to the inhibitory effect of ABA on seed germination is enhanced. Consistent with this finding, we demonstrate that the ABA-responsive gene, RAB18, is hyperinduced in seeds imbibed in ABA plus PAO, compared with seeds imbibed only with ABA. PMID- 16211389 TI - A phytochrome-dependent embryonic factor modulates gibberellin responses in the embryo and micropylar endosperm of Datura ferox seeds. AB - The promotion of germination by phytochrome is associated with extensive changes both in the embryo and in the micropylar region of the endosperm (ME) of Datura ferox seeds. These changes require de novo gibberellins (GAs) biosynthesis in the embryo, the site where the light stimulus is perceived. GAs stimulate embryo growth potential and move to ME, promoting the expression of genes related with weakening. We report here that, in addition, phytochrome stimulates the sensitivity of the seeds to gibberellic acid (GA). The phytochrome-induced signal is produced in the embryo and enhances the stimulus by GA of embryo growth potential (EGP) and the promotion of the expression of proteins thought to participate in ME weakening: endo-beta-mannanase (EC 3.2.1.78), endo-beta mannosidase (EC 3.2.1.25) and expansin. Our results suggest that the cytokinins may be a component of the embryonic signal. Phytochrome also modulates DfPHOR and DfMYB transcript levels in ME. These genes show a high identity with components of GAs signaling identified in other species. Expression of DfPHOR in the ME is apparently regulated by phytochrome through the supply of GAs from the embryo to ME, whereas DfMYB expression is regulated by an embryonic factor with some of the characteristics of the one that modulates seed sensitivity to GAs. PMID- 16211390 TI - Activation of Arabidopsis MAPK kinase kinase (AtMEKK1) and induction of AtMEKK1 AtMEK1 pathway by wounding. AB - We have constructed a series of deletion mutants of Arabidopsis MAPK kinase kinase (AtMEKK1) and obtained a constitutively active mutant, AtMEKK1Delta166, which lacks in self-inhibitory sequence of N-terminal 166 amino acids but still has substrate specificity. AtMEKK1Delta166 predominantly phosphorylates AtMEK1, an Arabidopsis MAPKK, but not its double mutant (AtMEK1T218A/S224E), suggesting that Thr-218 and Ser-224 are the phosphorylation sites. In wounded seedlings, AtMEKK1 was activated and phosphorylated its downstream AtMEK1. Furthermore, analysis using anti-AtMEKK1 and anti-AtMEK1 antibodies revealed that the interaction between the two proteins was signal dependent. These results suggest the presence of AtMEKK1-AtMEK1 pathway induced by wounding. PMID- 16211391 TI - Calcification of myocardial necrosis is common in mice. AB - Independent of the severity, phenotypes and clinical outcomes of myocardial infarction may vary considerably in patients, suggesting a strong genetic influence on healing and adaptive processes. Since little is known about these genetic determinants, we examined the tissue response to myocardial injury in seven inbred mouse strains, including those employed for gene targeting or transgenic overexpression. Myocardial necrosis was produced by non-ischemic, trans-diaphragmal freeze-thaw injury in strains C57BL/6, C3H/He, DBA/2, BALB/c, 129S1, FVB/n and A/J. Two days after injury, necrotic cardiomyocytes calcified in C3H/He, DBA/2, BALB/c and 129S1, a phenotype known as dystrophic cardiac calcinosis (DCC). The susceptibility to DCC of 129S1 was determined by Dyscalc1, a locus on chromosome 7, which was identified previously in C3H/He and DBA/2. DCC was also observed in C3H/He following ischemic injury by permanent coronary artery ligation, indicating that DCC was independent of the mode of injury. In contrast, strains C57BL/6, FVB and A/J were resistant to DCC, showing formation of a fibrous scar without calcification. The development of DCC was studied in detail in C3H/He and C57BL/6. In both strains, no calcium deposition and only little structural disintegration were noted in necrotic myocardium 24 h after injury upon calcium-sensitive fluorescence staining. Ultrastructural examination revealed calcified mitochondria in C3H/He that may have served later as a nidus for rapid intracellular calcification of cardiomyocytes. We concluded that the susceptibility to calcification of myocardial necrosis may be common among inbred strains and should be recognised as a strong genetic modifier of experimental myocardial injury. PMID- 16211392 TI - Ultrastructural analysis of mouse embryonic stem cell-derived chondrocytes. AB - Pluripotent embryonic stem (ES) cells cultivated as cellular aggregates, so called embryoid bodies (EBs), differentiate spontaneously into different cell types of all three germ layers in vitro resembling processes of cellular differentiation during embryonic development. Regarding chondrogenic differentiation, murine ES cells differentiate into progenitor cells, which form pre-cartilaginous condensations in the EB-outgrowths and express marker molecules characteristic for mesenchymal cell types such as Sox5 and Sox6. Later, mature chondrocytes appear which express collagen type II, and the collagen fibers show a typical morphology as demonstrated by electron-microscopical analysis. These mature chondrogenic cells are organized in cartilage nodules and produce large amounts of extracellular proteoglycans as revealed by staining with cupromeronic blue. Finally, cells organized in nodules express collagen type X, indicating the hypertrophic stage. In conclusion, differentiation of murine ES cells into chondrocytes proceeds from the undifferentiated stem cell via progenitor cells up to mature chondrogenic cells, which then undergo hypertrophy. Furthermore, because the ES-cell-derived chondrocytes did not express elastin, a marker for elastic cartilage tissue, we suggest the cartilage nodules to resemble hyaline cartilage tissue. PMID- 16211393 TI - Gene expression profiling in rice young panicle and vegetative organs and identification of panicle-specific genes through known gene functions. AB - In rice, at the stage from pistil and stamen primordia formation to microsporocyte meiosis, the young panicle organs (YPO) make a great contribution to grain productivity. This period corresponds to the onset of meiosis and marks the transition from vegetative to reproductive stages. By comparing gene expression profiling of YPO with that of rice aerial vegetative organs (AVO), it is possible to gain further molecular insight into this period that is developmentally and functionally important. In this report, a total of 92,582 high-quality ESTs from 5'-end sequencing, including 44,247 from YPO and 48,335 from AVO, were obtained and classified. There were 12,884 (29.12%) ESTs from YPO and 16,304 (33.73%) ESTs from AVO matched to known genes, which generated 1,667 and 2,172 known genes, respectively, after integration of these ESTs. From the functions of known homologous genes, we identified some tissue- and developmental stage-specified genes in YPO. The expression of these genes clearly reflected the unique functional characteristics of YPO. Furthermore, we estimated that there are about 10,000 mRNAs specifically expressed in rice YPO. PMID- 16211394 TI - Root distribution and interactions between intercropped species. AB - Even though ecologists and agronomists have considered the spatial root distribution of plants to be important for interspecific interactions in natural and agricultural ecosystems, few experimental studies have quantified patterns of root distribution dynamics and their impacts on interspecific interactions. A field experiment was conducted to investigate the relationship between root distribution and interspecific interactions between intercropped plants. Roots were sampled twice by auger and twice by the monolith method in wheat (Triticum aestivum L.)/maize (Zea mays L.) and faba bean (Vicia faba L.)/maize intercropping and in sole wheat, maize, and faba bean up to 100 cm depth in the soil profile. The results showed that the roots of intercropped wheat spread under maize plants, and had much greater root length density (RLD) at all soil depths than sole wheat. The roots of maize intercropped with wheat were limited laterally, but had a greater RLD than sole-cropped maize. The RLD of maize intercropped with faba bean at different soil depths was influenced by intercropping to a smaller extent compared to maize intercropped with wheat. Faba bean had a relatively shallow root distribution, and the roots of intercropped maize spread underneath them. The results support the hypotheses that the overyielding of species showing benefit in the asymmetric interspecific facilitation results from greater lateral deployment of roots and increased RLD, and that compatibility of the spatial root distribution of intercropped species contributes to symmetric interspecific facilitation in the faba bean/maize intercropping. PMID- 16211395 TI - p53 Status does not affect photodynamic cell killing induced by hypericin. AB - PURPOSE: Given that p53 is a tumor suppressor that plays a central role in the cellular response to DNA damage and that more than 50% of all cancers have mutated p53, the wider utility of photodynamic therapy (PDT) in the treatment of cancer will depend on an understanding of whether p53 status modulates response to PDT. In this study, we investigated the photosensitivity of isogenic cell lines that differ only in their p53 status to PDT using hypericin as the photosensitizer. METHODS: Acute (MTT) and chronic (clonogenic) cytotoxic assays were performed on two osteosarcoma cell-lines (U2OS and U2OS+p53DD) that are isogenic except that the latter expresses dominant negative p53. The inducible expression of p53 was determined on western blots. Uptake of hypericin, cell cycle profile analysis, measurement of membrane phosphatidylserine externalization and changes in mitochondrial membrane potential were investigated using flow cytometry. RESULTS: Hypericin uptake was observed to be equivalent in U2OS and U2OS+p53DD cells. There were no significant differences in cell killing between these cell-lines in both the MTT and clonogenic assays (IC(50) of 0.4 microg/ml from MTT assay). p53 expression did not increase up to 24 h after PDT treatment in both cell lines. There were also no significant differences in the cell-cycle arrest profiles and timing of onset of apoptosis. CONCLUSIONS: Taken together, these results suggest that the status of p53 may not be important in PDT-mediated cell killing or induction of apoptosis. By extension, these results imply that PDT may be used with equal efficacy for the treatment of p53-positive and -negative tumors. PMID- 16211396 TI - Treatment of hyperlipidemia associated with Niemann-Pick disease type B by fenofibrate. PMID- 16211397 TI - Absence of breast-feeding is associated with the risk of type 1 diabetes: a case control study in a population with rapidly increasing incidence. AB - There are indications that the effect of environmental factors on the risk of type 1 diabetes mellitus (T1DM) is increasing over time. This can be documented by the rapid increase of T1DM incidence in genetically stable populations. Our aim was to study an association of T1DM with the variable factors of the perinatal period and of early infancy, using data from children born over a period of changing exposure to some of the studied factors. A case-control dataset was analysed, consisting of 868 diabetic children and 1,466 anonymous controls, mostly schoolmates of the children with T1DM. The data were collected using structured questionnaires completed by parents. After performing univariate analyses, the associations were analysed using multiple logistic regression adjusted for potential confounders, including the year of birth. The risk of T1DM decreased with increasing duration of breast-feeding, while no breast-feeding was associated with an increased T1DM risk, OR=1.93 [95% CI: 1.33-2.80], breast feeding for more than 12 months was protective, OR=0.42 [95% CI: 0.22-0.81], both being relative to the reference category of breast-feeding for 1-3 months. A short duration of day-care attendance (none or less than 1 year) was weakly associated with the risk of T1DM, OR=1.65 [95% CI: 1.05-2.62]. No association was detected between T1DM and signs of prenatal infections, perinatal stress factors, birth size and weight, indicators of crowding or the presence of a domestic pet in the household. Short breast-feeding period and short attendance to day care is associated with the risk of T1DM in Czech children. PMID- 16211398 TI - Status epilepticus due to attempted suicide with isoniazid. PMID- 16211399 TI - The contribution of pulse oximetry to the early detection of congenital heart disease in newborns. AB - Approximately half of all newborns with congenital heart disease are asymptomatic in the first few days of life. Early detection of ductal-dependant cardiac malformations prior to ductal closure is, however, of significant clinical importance, as the treatment outcome is related to the time of diagnosis. Pulse oximetry has been proposed for early detection of congenital heart disease. The aims of the present study were: 1) to determine the effectiveness of a pulse oximetric screening performed on the first day of life for the detection of congenital heart disease in otherwise healthy newborns and 2) to determine if a pulse-oximetric screening combined with clinical examination is superior in the diagnosis of congenital heart disease to clinical examination alone. This is a prospective, multi-centre study. Postductal pulse oximetry was performed between six and twelve hours of age in all newborns of greater than 35 weeks gestation. If pulse-oximetry-measured arterial oxygen saturation was less than 95%, echocardiography was performed. Pulse oximetry was performed in 3,262 newborns. Twenty-four infants (0.7%) had repeated saturations of less than 95%. Of these infants, 17 had congenital heart disease and five of the remaining seven had persistent pulmonary hypertension. No infant with a ductal-dependant or cyanotic congenital heart disease exhibited saturation values greater or equal to 95%. CONCLUSION: postductal pulse-oximetric screening in the first few days of life is an effective means for detecting cyanotic congenital heart disease in otherwise healthy newborns. PMID- 16211400 TI - Congenital hyperekplexia: five sporadic cases. AB - We report fives sporadic cases of hyperekplexia or startle disease characterized by a highly exaggerated startle reflex and tonic attacks. Affected neonates suffer from prolonged periods of stiffness and are at risk for sudden death from apnea. An early diagnosis is needed. Sudden loud sounds, unexpected tactile stimuli or percussion at the base of the nose can also elicit excessive jerking or tonic attack. The diagnosis of hyperekplexia is a purely clinical one. A defect of the alpha1 subunit of inhibitory glycine receptor (GLRA1) has been observed in the dominant form with a mutation in the chromosome 5. Clonazepam is effective and decreases the severity of the symptoms. The disease tends to improve after infancy and the psychomotor development is normal. The major form of "hyperekplexia" should be considered whenever one is confronted with neonatal hypertonicity associated with paroxysmal tonic manifestations (without electroencephalography anomalies). CONCLUSION: the diagnosis of hyperekplexia should be evaluated in any neonate with tonic attacks without evident cause. PMID- 16211401 TI - Plagiocephalometry: a non-invasive method to quantify asymmetry of the skull; a reliability study. AB - Deformational plagiocephaly (DP) in newborns and very young children is a common problem in daily practice. The intrarater and interrater reliability of plagiocephalometry (PCM), a new, non-invasive, inexpensive instrument to assess and quantify the asymmetry of the skull, is evaluated at the outpatient Department of Physical Therapy of the Bernhoven Hospital at Veghel, The Netherlands. Using a thermoplastic material to mould the outline of the infant's skull, a reproduction of the skull shape is performed on paper, allowing for accurate cephalometric measurements. Fifty children (aged 0-24 months), with or without positional preference of the head, and with or without DP, were measured three times by two separate, experienced pediatric physical therapists. Intraclass correlation coefficients (ICC) regarding the measurements of the drawn lines were all above 0.92 (intrarater reliability) and 0.90 (interrater reliability). The ICCs of the plagiocephaly indicators ear deviation (ED), antero sinistra-antero-dextra (ASAD), postero-dextra-postero-sinistra (PDPS) and oblique diameter difference (ODD) were 0.88, 0.57, 0.92 and 0.96, respectively, for the intrarater reliability and 0.90, 0.65, 0.94 and 0.96, respectively, for the interrater reliability. The ICCs of the two indices oblique diameter difference index (ODDI) and cranial proportional index (CPI) were 0.97 and 0.96, respectively, for the intrarater reliability and 0.95 and 0.92, respectively, for the interrater reliability. The limits of agreement according to Bland Altman, comprising 95% of the differences between two measurements (2 sd), were 4.3 mm (ED), 5.9 mm (ASAD), 3.0 mm (PDPS), 3.4 mm (ODD), 2.7% (ODDI) and 4.5% (CPI) for the intrarater reliability, and 3.7 mm (ED), 5.2 mm (ASAD), 2.4 mm (PDPS), 3.3 mm (ODD), 2.9% (ODDI) and 5.8% (CPI) for the interrater reliability. CONCLUSION: We conclude that PCM is an easy-to-apply, non-invasive and reliable measurement instrument to assess skull asymmetry with good clinical accuracy and low application costs. PCM might serve as an instrument to be used in all levels of care for children with DP, and might provide information concerning the natural course of DP, as well as the assessment of the effects of conservative treatment strategies on DP. PMID- 16211403 TI - Alamethicin influence on the membrane bending elasticity. AB - We investigate the bending elasticity of lipid membranes with the increase of the alamethicin concentrations in the membrane via analysis of the thermally induced shape fluctuations of quasi-spherical giant vesicles. Our experimental results prove the strong influence of alamethicin molecules on the bending elasticity of diphytanoyl phosphatidylcholine and dilauroyl phosphatidylcholine membranes even in the range of very low peptide concentrations (less than 10(-3) mol/mol in the membrane). The results presented in this work, testify to the peripheral orientation of alamethicin molecules at low peptide concentrations in the membrane for both types of lipid bilayers. An upper limit of the concentration of the peptide in the membrane is determined below which the system behaves as an ideal two-dimensional solution and the peptide molecules have a planar orientation in the membrane. PMID- 16211402 TI - Mechanisms of iron-sulfur cluster assembly: the SUF machinery. AB - Biosynthesis of iron-sulfur clusters is a cellular process which depends on complex protein machineries. Escherichia coli contains two such biosynthetic systems, ISC and SUF. In this review article we specifically make a presentation of the various Suf proteins and discuss the molecular mechanisms by which these proteins work together to assemble Fe and S atoms within a scaffold and to transfer the resulting cluster to target proteins. PMID- 16211404 TI - [Cerebral secondary prevention--clopidogrel alone or in combination with ASS]. PMID- 16211405 TI - [Diagnostics and therapy for invasive fungal infections in an intensive care unit]. AB - Invasive fungal infections are associated with a high mortality and have been increasing in incidence over the last few decades. Candidemia and, less commonly, invasive pulmonary aspergillosis are the most relevant fungal infections in critical care medicine. Risk factors for systemic Candida infections are the use of broad-spectrum antibiotics, a prolonged stay in an intensive care unit and gastrointestinal injury or surgery. Invasive aspergillosis usually occurs in immunocompromised patients. The diagnosis of invasive fungal infections remains challenging. The therapeutic spectrum includes fluconazol, conventional and liposomal amphotericin B, and the recently introduced agents caspofungin and voriconazol. For rational and cost-effective use, the clinician requires precise knowledge of the indications and limitations of these agents. This review focuses on the diagnostic and therapeutic options in severe Candida infections and invasive aspergillosis. PMID- 16211406 TI - Are organic cation transporters capable of transporting prostaglandins? AB - The non-neuronal monoamine transporters OCT1, OCT2 and EMT (human gene symbols SLC22A1-A3) efficiently transport a number of positively-charged monoamines and some small organic cations across the plasma membrane, and thus are implicated in the inactivation of released monoamine transmitters (e.g. noradrenaline, histamine, agmatine) in vivo. Although prostaglandins are full anions at physiological pH, data from a recent publication suggest efficient transport of the prostaglandins PGE2 and PGF2alpha by OCT1 and OCT2. In the present study we have reexamined transport of PGE2 by OCT2 from human (OCT2h). Uptake of substrate into monolayers of 293 cells, stably transfected to express OCT2h, was compared to uptake into non-transfected control cells. Efficiency of transport of the established substrate 3H-1-methyl-4-phenylpyridinium (MPP+), expressed as clearance, was high at 81 microl min(-1) mg protein(-1) on average. By contrast, uptake of 3H-PGE2 was virtually identical for control cells and OCT2h cells. The efficiency of transport was 0.1+/-0.6, 1.0+/-0.3, and 0.7+/-0.4 microl min(-1) mg protein(-1) for cell lysis with methanol, HClO4, and Triton X-100 respectively. Similar results were obtained with unlabeled MPP+ (192+/-12 microl min(-1) mg protein(-1)) and PGE2 (0.3+/-0.1 microl min(-1) mg protein(-1)) in LC-MS/MS analysis. We conclude that OCT2h is not capable of transporting prostaglandins. The data from the previous report may represent binding rather than transport. Our comparison of transport efficiencies confirms the notion that relevant substrates of OCT1, OCT2, and EMT must carry a positive charge. PMID- 16211407 TI - Gene expression studies in prostate cancer tissue: which reference gene should be selected for normalization? AB - Using quantitative reverse transcription-polymerase chain reaction (RT-PCR), reference genes are utilized as endogenous controls for relative quantification of target genes in gene profiling studies. The suitability of housekeeping genes for that purpose in prostate cancer tissue has not been sufficiently investigated so far. The objective of this study was to select from a panel of 16 potential candidate reference genes the most stable genes for gene normalization. Expression of mRNA encoding ACTB, ALAS1, ALB, B2M, G6PD, GAPD, HMBS, HPRT1, K ALPHA-1, POLR2A, PPIA, RPL13A, SDHA, TBP, UBC, and YWHAZ was examined in matched, microdissected malignant and nonmalignant tissue specimens obtained from 17 nontreated prostate carcinomas after radical prostatectomy by real-time RT-PCR. The genes studied displayed a wide expression range with cycle threshold values between 16 and 37. The expression was not different between samples from pT2 and pT3 tumors or between samples with Gleason scores <7 and >or=7 (P>0.05). ACTB, RPL13A, and HMBS showed significant differences (P<0.02 at least) in expressions between malignant and nonmalignant pairs. All other genes did not differ between the matched pairs, and the software programs geNorm and NormFinder were used to ascertain the most suitable reference genes from these candidates. HPRT1, ALAS1, and K-ALPHA-1 were calculated by both programs to be the most stable genes covering a broad range of expression. The expression of the target gene RECK normalized with HRPT1 alone and with the normalization factors generated by the combination of these three reference genes as well as with the unstable genes ACTB or RPL13A is given. That example shows the significance of using suitable reference genes to avoid erroneous normalizations in gene profiling studies for prostate cancer. The use of HPRT1 alone as a reference gene shown in our study was sufficient, but the normalization factors generated from two (HRPT1, ALAS1) or all three genes (HRPT1, ALAS1, K-ALPHA-1) should be considered for an improved reliability of normalization in gene profiling studies of prostate cancer. PMID- 16211408 TI - Agrobacterium and biolistic transformation of onion using non-antibiotic selection marker phosphomannose isomerase. AB - A new selection system for onion transformation that does not require the use of antibiotics or herbicides was developed. The selection system used the Escherichia coli gene that encodes phosphomannose isomerase (pmi). Transgenic plants carrying the manA gene that codes for pmi can detoxify mannose-6-phosphate by conversion to fructose-6-phosphate, an intermediate of glycolysis, via the pmi activity. Six-week-old embryogenic callus initiated from seedling radicle was used for transformation. Transgenic plants were produced efficiently with transformation rates of 27 and 23% using Agrobacterium and biolistic system, respectively. Untransformed shoots were eliminated by a stepwise increase from 10 g l(-1) sucrose with 10 g l(-1) mannose in the first selection to only 10 g l(-1) mannose in the second selection. Integrative transformation was confirmed by PCR, RT-PCR and Southern hybridization. PMID- 16211409 TI - Effect of tumor necrosis factor alpha and vascular permeability growth factor on albuminuria in rats. AB - The purposes of this study were to measure the serum levels of vascular permeability growth factor (VPGF) and tumor necrosis factor alpha (TNFalpha) in minimal lesion nephrotic syndrome (MLNS) patients and to assess their effect on albuminuria in rats. Serum for VPGF and TNFalpha was obtained during relapse and remission from 18 MLNS patients. Tumor necrosis factor alpha was infused at the rate of 10 and 20 ng/h and VPGF at the rate of 20 and 40 ng/h for 5 days into the left renal artery of rats. Urinary albumin (24-h collection) was measured prior to infusion and on days 2, 4 and 5. Rats infused with 1% bovine serum albumin served as controls. Serum VPGF and TNFalpha levels in MLNS patients in relapse were not different from those seen during remission. A significant increase in albuminuria was observed on day 4 and 5 only when rats were infused with TNFalpha at the rate of 20 ng/h as compared to the excretion seen in same animals prior to the infusion of cytokine and on days 4 and 5 of normal controls. Neither VPGF nor TNFalpha seems to be the circulating pathogenic cytokine for proteinuria in MLNS. However, TNFalpha may contribute to the increased albuminuria via a paracrine effect at the glomerulus. PMID- 16211410 TI - Tau and Kappa effects in physical space: the case of audition. AB - The organization of spatio-temporal information in an auditory memory task was studied in two experiments. Stimuli consisted of four different configurations of eight sequentially presented beeps. In two configurations, the stimuli were space time congruent, with (constant or variable) inter-stimulus distances corresponding to (constant or variable) inter-stimulus time intervals. In the other two configurations, the stimuli were space-time incongruent, with (constant or variable) inter-stimulus distances not corresponding to (variable or constant) inter-stimulus time intervals. After a learning phase consisting of 20 presentations of the target configuration, participants reproduced the spatial (Experiment 1) or temporal (Experiment 2) characteristics of the target 60 times in succession without re-examining the target configuration. Accuracy (with respect to the target) and variability (between responses) were found to evolve independently. In the incongruent space-time conditions, effects of variable inter-stimulus time intervals or distances on the reproduction of, respectively, constant distances (Tau effect) or constant time intervals (Kappa effect) were observed, while the reverse was not the case. Thus, dimensional interference occurred when the dimension to be ignored was variable. The results are discussed in the light of the distinction between properties of the stabilized mental image and the process of stabilization. PMID- 16211412 TI - Co-occurrence of Naobranchia lizae (Copepoda) and Metamicrocotyla macracantha (Monogenea), gill parasites of the striped mullet Mugil cephalus. AB - The copepod Naobranchia lizae (Kroyer, 1863) and the monogenean Metamicrocotyla macracantha (Alexander, 1954; Koratha, 1955) are gill parasites found on the striped mullet (Mugil cephalus) in the Charleston Harbor Estuarine System (South Carolina, USA). Branchial distribution of each species was studied in mono- and bispecific conditions. No preference for the right or left side of the fish host was observed for either species in either condition. Both species exhibited heterogeneous distribution among the gill arches, with a preference for arch I. N. lizae exhibited intraspecific competition and a saturation threshold in both mono- and bispecific conditions. A shift in niche toward the posterior arches was observed for M. macracantha only in bispecific infection. Interspecific competition was detected exclusively on the posterior arches, where M. macracantha seemed out-competed by N. lizae. The data indicated that both neutral and negative interactions played a role in determining the distribution of N. lizae and M. macracantha individuals on the gill arches of M. cephalus. PMID- 16211411 TI - [Sleep-related breathing disorders. Sleep anamnesis questionnaire and determination of clinical results within the framework of staged diagnostics]. AB - Sleep-related breathing disorders (SBAS) affect approximately 8% of the population of middle aged adults. At the age of 20 years, approximately 10% of the population snore, while at the age of 60 about 50% of men snore. Some 9% of middle aged women and 24% of middle aged men have an apnea hypopnea index (AHI) of >5 (number of nocturnal apnea and hypopnea per hour of sleep). Sleep apnea hypopnea syndrome is found in 2% of the women and 4% of the men, i.e. they have an AHI>5 associated with daytime sleepiness. Forms, check lists, summaries and patient-readable questionnaires have proved helpful in the evaluation of SBAS. PMID- 16211413 TI - Characterisation of carbohydrate-binding sites in developmental stages of Myxobolus cerebralis. AB - Glycans and lectins (carbohydrate-binding molecules) form a mutual recognition system, which enables parasitic organisms to attach themselves to the host cells and/or take part in the migration of their developmental stages into the target tissue. The aim of the present study was to identify and characterise the potential binding activity of glycoconjugates in different developmental stages of Myxobolus cerebralis, the causative agent of whirling disease in salmonids. The binding patterns of 13 biotinylated neoglycoconjugates were histochemically examined in thin-sections of infected rainbow trout (Oncorhynchus mykiss) and oligochaetes (Tubifex tubifex), as well as isolated waterborne triactinomyxon spores. A distinct structure-selective and developmental stage-regulated expression of certain classes of carbohydrate binding was observed. In triactinomyxon spores, the expression of carbohydrate binding activity for alpha l-Fuc-BSA-biotin, alpha-d-GalNAc-BSA-biotin, beta-d-GlcNAc-BSA-biotin, Lac-BSA biotin and ASF-biotin was up-regulated in the polar capsules; the shell valves showed no activity. In the gut of T. tubifex, polar capsules of the parasite showed strong positive reaction only for beta-d-GlcNAc-BSA-biotin. In fish cartilage, polar capsules were negative, but the spore shell valves showed a broad range of carbohydrate binding activity. No activity was detected for either alpha6- or alpha3-linked N-acetyl-d-neuraminic acid to galactose. An adhesion assay was performed on GlycoWell plates and Myxobolus spores were found to specifically adhere to matrices containing residues of lactose, fucose, galactose, N-acetyl-d-galactosamine and N-acetyl-d-glucosamine. This is the first study to identify lectin activity in a myxozoan parasite; activity that is likely to play a role in the recognition systems involved in host specificity and the processes of spore attachment and invasion. PMID- 16211414 TI - Calcification of Trichinella spiralis larval capsule. AB - Trichinella spiralis larvae were examined in TEM to identify calcareous corpuscles in the outer part of parasite capsule. The microroentgenographic analysis of calcareous corpuscles mainly demonstrated the presence of phosphorus and calcium. The physiological importance of calcareous corpuscles, as well as their significance in decay of T. spiralis larvae were discussed. PMID- 16211416 TI - Neonatal gut injury and infection rate: impact of surgical debridement on outcome. AB - Infectious burden of gut injury (G-INJ) associated with necrotizing enterocolitis (NEC) or with spontaneous intestinal perforation (SIP) in neonates has not been ascertained. We sought to test the hypotheses that: (1) infants with G-INJ develop higher number of infections including non-concurrent infections than infants without G-INJ in a neonatal intensive care unit (NICU); (2) surgical debridement (DEB) of infants with severe G-INJ is associated with lower infectious morbidity and mortality. All infants admitted to the regional NICU from October 1991 to February 2003 were included in this prospective prevalence investigation of G-INJ and infections. Non-viable (<23 week gestational age) infants, infants with congenital anomalies, and those who developed NEC after SIP were excluded. Standard definitions of National Centers for Disease Control and Prevention were used for different categories of infections. Episodes of infections were classified as concurrent or non-concurrent (post G-INJ) based upon their timing in association with G-INJ. Infants with G-INJ associated with Bell stage II or higher NEC or with SIP were further stratified by DEB into two subgroups. A previously described 7-point clinical score was used to divide G-INJ into mild (0-2), moderate (3-5), and severe (6-7) categories. Surgical outcomes were determined by using chi(2) and logistic regression analyses. Data are expressed as mean +/- SD or as odds ratio (OR) with 95% confidence intervals (CI); P < 0.05 was considered significant. Of all 5,481 infants, 954 (17.4%) developed 1,734 episodes of infections. Prevalence of G-INJ was 4% (n = 222); of these, 33% (n = 73) underwent DEB. Infants with G-INJ had lower mean birth weight (1,414+/-766 vs. 2,153+/-104 g; P < 0.0001) and lower mean gestational age (29.6+/-4.2 vs. 32.9+/-4.8 weeks; P < 0.0001) than their peers (n = 5,259). Controlling for birth weight and gestational age, odds for non-concurrent blood stream infections (BSIs) in G-INJ infants were higher (OR 13.98, CI 10.289-19.01, P < 0.0001) than the remaining population without G-INJ. Forty-four percent of all episodes of fungemia, 32% of all episodes of BSIs occurred in G-INJ infants (P < 0.0001). Within the G-INJ group, there were no demographic differences between the DEB and non-DEB infants. Controlling for severity of G-INJ, odds for non-concurrent BSIs (OR 3.45, CI 1.04-11.36, P < 0.05) and for mortality (OR 3.35, CI 1-10, P < 0.05) among non-DEB infants were higher than in DEB infants. Infants with G-INJ suffered from a disproportionate number of all blood-stream infections in our intensive care nursery. Infants with severe G-INJ whose management includes DEB are more likely to survive and to incur less infectious morbidity. PMID- 16211417 TI - Atrichia with papular lesions in two Pakistani consanguineous families resulting from mutations in the human hairless gene. AB - Atrichia with papular lesions (APL) is a rare autosomal recessive form of total alopecia, characterized by hair loss soon after birth and the development of papular lesions of keratin-filled cysts over extensive areas of the body. Mutations in the hairless (hr) gene, a putative single zinc finger transcription factor, have been implicated in the pathogenesis of this disorder. In the present study, we describe two novel deletion mutations in exons 2 and 8 of the human hairless gene leading to frameshift and downstream premature termination codons in two consanguineous Pakistani families affected with atrichia. PMID- 16211415 TI - Serine proteinases of the human body louse (Pediculus humanus): sequence characterization and expression patterns. AB - After the previous characterization of one trypsin gene (Try1) of the human body louse Pediculus humanus, genes encoding a second trypsin (Try2) and a chymotrypsin (Chy1) have been cloned using degenerate serine proteinase primers and 5'- and 3'-RACE, and sequenced. The deduced 259 and 267 amino acid sequences of Try2 and Chy1 show an identity of 33%-40% to trypsinogens and chymotrypsinogens of other insects. Considering previously published partial sequences, P. humanus possesses at least one Try1 gene, five variants/isoforms of Try2 and six variants/isoforms of Chy1. The genomic DNA of Try2 contains three introns and Chy1 contains five introns. Using whole mount in situ hybridization, gene expression of Try1, Try2 and Chy1 has been localized not only in the distensible anterior region of the midgut of lice but sometimes also in the area following the distensible region. The Try2 gene was always expressed at much lower levels than Try1 or Chy1. This lower expression, the constitutive expression of Try1 and Chy1 at 1, 2, 6, 12 and 24 h after feeding of adults and the regional differences have been verified in quantitative real-time PCR. PMID- 16211419 TI - Carpals and epiphyses of radius and ulna as age indicators. AB - Estimation of skeletal age using radiographic images is widely used for assessing biological growth in clinical and auxological studies. The most frequent areas used for age estimation in children and adolescents are tooth and wrist/hand, both giving good results with only a low level of radiation. In particular, ossification of the carpals shows good agreement with chronological age. This study of a sample of 150 Italian children and adolescents aged between 5 and 17 years focused on analyzing the possible applications of the proportion of carpal area (Ca) mineralization as a criterion of age estimation. The ratio between the total area of carpal bones and epiphyses of the ulna and radius (Bo) and Ca was calculated. This ratio (Bo/Ca) was used for linear regression analysis. The regression model, describing age as a linear function of the ratio Bo/Ca, yielded the following equation: Age=-3.253+0.719 g+20.610 Bo/Ca, and explained 83% of the total variance (R (2)=0.83). The median of the absolute values of residuals (observed age minus predicted age) was 0.08 years, with a quartile deviation of 1.59 years, and a standard error of estimate of 1.19 years. PMID- 16211421 TI - Evaluation of early pelvic floor physiotherapy on the duration and degree of urinary incontinence after radical retropubic prostatectomy in a non-teaching hospital. AB - The objective of this study was to study the effect of early pelvic floor re education on the degree and duration of incontinence and to evaluate the results of radical retropubic prostatectomy (RRP) performed in a non-teaching hospital. This is a non-randomised study. From March 2000 to November 2003, 57 consecutive men, who underwent RRP for localized prostate cancer, participated in a pelvic floor re-educating program. Continence was defined as a loss of no more than 2-g urine on the 24-h pad test and no use of pads. The 24-h pad test was performed once in every 4 weeks until the patient indicated that he was continent. Diurnal and nocturnal continence was achieved after 1, 2, 3, 6 and 12 months post catheter removal in 40, 49, 70, 86 and 88% of all men, respectively. Comparison of our results with current literature suggest that the time period towards continence after a RRP can be shortened relevantly if pelvic floor re-education is started directly after catheter removal. PMID- 16211422 TI - Translevator posterior intravaginal slingplasty: anatomical landmarks and safety margins. AB - The posterior intravaginal sling is a new tension-free needle suspension technique. It is used for the treatment of middle compartment (vaginal vault or uterine) prolapse. The Prolene sling suspends the vagina at the upper border of level II support as described by DeLancey (Am J Obstet Gynecol 166:1717, 1992). Human cadaveric dissections were undertaken to explore the pertinent anatomy that is involved when using this blind needle technique. Pre-dissected cadaveric material was used to obtain didactic illustrations of the anatomy of the procedure. Description of the surgical technique using anatomical landmarks and relative distances of the needle to these landmarks will improve the surgeon's visual understanding of the procedure. The measurements obtained demonstrate that the needle stays at a minimal distance of 4 cm away from the major (pudendal) vessels that could potentially cause life-threatening haemorrhage. PMID- 16211420 TI - The human brain and its neural stem cells postmortem: from dead brains to live therapy. AB - Contrary to the traditional dogma of being a relatively invariable and quiescent organ lacking the capability to regenerate, there is now widespread evidence that the human brain harbors multipotent neural stem cells, possibly throughout senescence. These cells can divide and give rise to neuroectodermal progeny in vivo and are now regarded as powerful prospective candidates for repairing or enhancing the functional capability of neural tissue in trauma or diseases associated with degeneration or malperfusion. Hopes primarily rest upon techniques to either recruit endogenous stem cells or to utilize exogenous donor derived material for transplantation. In the search for suitable human cell sources, embryonic, fetal, and adult stem cells appear highly controversial, as they are accompanied by various still-unresolved moral and legal challenges. Fascinatingly, however, recent reports indicate the successful isolation and expansion of viable neural stem cells from the rodent and human brain within a considerable postmortem interval, suggesting that postmortem neural stem cells could potentially become an acceptable alternative cellular resource. This article will provide a brief overview about neural stem cells, their prominent features, and prospects for a cellular therapy, and will furthermore illuminate the cells in particular with respect to their newly discovered postmortem provenience, their advantage as a potential cell source, and several unfolding forensic considerations. Also, important ethical, social, and legal implications arising from this hitherto unpracticed cellular harvest of brain tissue from the deceased are outlined. PMID- 16211423 TI - Haplotype analysis of the human endogenous retrovirus locus HERV-K(HML-2.HOM) and its evolutionary implications. AB - We and others recently identified an almost-intact human endogenous retrovirus (HERV), termed HERV-K(HML-2.HOM), that is usually organized as a tandem provirus. Studies on HERV proviral loci commonly rely on the analysis of single alleles being taken as representative for a locus. We investigated the frequency of HERV K(HML-2.HOM) single and tandem alleles in various human populations. Our analysis revealed that another HERV-K(HML-2) locus, the so-called HERV-K(II) provirus, is also present as a tandem provirus allele in the human population. Proviral tandem formations were identified in various nonhuman primate species. We furthermore examined single nucleotide polymorphisms (SNPs) within the HERV-K(HML-2.HOM) proviral gag, prt, and pol genes, which all result in nonsense mutations. We identified four proviral haplotypes displaying different combinations of gag, prt, and pol SNPs. Haplotypes harboring completely intact proviral genes were not found. For the left provirus of the tandem arrangement a haplotype displaying intact gag and prt genes and a mutated pol was found in about two-thirds of individuals from different ethnogeographic origins. The same haplotype was always found in the right provirus. The various haplotypes point toward multiple recombination events between HERV-K(HML-2.HOM) proviruses. Based on these findings we derive a model for the evolution of the proviral locus since germ line integration. PMID- 16211424 TI - Phylogeny of the Centrohelida inferred from SSU rRNA, tubulins, and actin genes. AB - Amoeboid protists are major targets of recent molecular phylogeny in connection with reconstruction of global phylogeny of eukaryotes as well as the search for the root of eukaryotes. The Centrohelida are one of the major groups of Heliozoa, classified in the Actinopodida, whose evolutionary position is not well understood. To clarify the relationships between the Centrohelida and other eukaryotes, we sequenced SSU rRNA, alpha-tubulin, and beta-tubulin genes from a centroheliozoan protist, Raphidiophrys contractilis. The SSU rRNA phylogeny showed that the Centrohelida are not closely related to other heliozoan groups, Actinophryida, Desmothoracida, or Taxopodida. Maximum likelihood analyses of the combined phylogeny using a concatenate model for an alpha- + beta-tubulin + actin data set, and a separate model for SSU rRNA, alpha- and beta-tubulin, and actin gene data sets revealed the best tree, in which the Centrohelida have a closer relationship to Rhodophyta than to other major eukaryotic groups. However, both weighted Shimodaira-Hasegawa and approximately unbiased tests for the concatenate protein phylogeny did not reject alternative trees in which Centrohelida were constrained to be sisters to the Amoebozoa. Moreover, alternative trees in which Centrohelida were placed at the node branching before and after Amoebozoa or Viridiplantae were not rejected by the WSH tests. These results narrowed the possibilities for the position of Centrohelida to a sister to the Rhodophyta, to the Amoebozoa, or to an independent branch between the branchings of Amoebozoa and Rhodophyta (or possibly Plantae) at the basal position within the bikonts clade in the eukaryotic tree. PMID- 16211425 TI - Molecular evolution of the plant virus family Bromoviridae based on RNA3-encoded proteins. AB - We have carried out an evolutionary study of the two proteins encoded by the RNA 3 from members of the plant virus family Bromoviridae. Using maximum likelihood methods, we have inferred the patterns of amino acid substitution that better explain the diversification of this viral family. The results indicate that the molecular evolution of this family was rather complex, with each protein evolving at different rates and according to different patterns of amino acid substitution. These differences include different amino acid equilibrium frequencies, heterogeneity in substitution rates among sites, and covariation among sites. Despite these differences, the model of protein evolution that better fits both proteins is one specifically proposed for the evolution of globular proteins. We also found evidence for coevolution between domains of these two proteins. Finally, our analyses suggest that the molecular clock hypothesis does not hold, since different lineages evolved at different rates. The implications of these results for the taxonomy of this important family of plant viruses are discussed. PMID- 16211426 TI - Mammalian GC content is very close to mutational equilibrium. PMID- 16211427 TI - Mitogenomic analyses place the gharial (Gavialis gangeticus) on the crocodile tree and provide pre-K/T divergence times for most crocodilians. AB - Based on morphological analyses, extant members of the order Crocodylia are divided into three families, Alligatoridae, Crocodylidae, and Gavialidae. Gavialidae includes one species, the gharial, Gavialis gangeticus. In this study we have examined crocodilian relationships in phylogenetic analyses of seven mitochondrial genomes that have been sequenced in their entirety. The analyses did not support the morphologically acknowledged separate position of the gharial in the crocodilian tree. Instead the gharial joined the false gharial (Tomistoma schlegelii) on a common branch that was shown to constitute a sister group to traditional Crocodylidae (less Tomistoma). Thus, the analyses suggest the recognition of only two Crocodylia families, Alligatoridae and Crocodylidae, with the latter encompassing traditional Crocodylidae plus Gavialis/Tomistoma. A molecular dating of the divergence between Alligatoridae and Crocodylidae suggests that this basal split among recent crocodilians took place approximately 140 million years before present, at the Jurassic/Cretaceous boundary. The results suggest that at least five crocodilian lineages survived the mass extinction at the KT boundary. PMID- 16211428 TI - Error minimization and coding triplet/binding site associations are independent features of the canonical genetic code. AB - The canonical genetic code has been reported both to be error minimizing and to show stereochemical associations between coding triplets and binding sites. In order to test whether these two properties are unexpectedly overlapping, we generated 200,000 randomized genetic codes using each of five randomization schemes, with and without randomization of stop codons. Comparison of the code error (difference in polar requirement for single-nucleotide codon interchanges) with the coding triplet concentrations in RNA binding sites for eight amino acids shows that these properties are independent and uncorrelated. Thus, one is not the result of the other, and error minimization and triplet associations probably arose independently during the history of the genetic code. We explicitly show that prior fixation of a stereochemical core is consistent with an effective later minimization of error. PMID- 16211431 TI - Study on Staphylococcus aureus strain HPC-250 for associated antibacterial property. AB - We isolated a Staphylococcus aureus strain HPC-250 producing antibacterial agent against Paenibacillus strain HPC-251. Both strains were isolated from the same environmental niche. The bacteria were identified using the partial sequencing of their TA-cloned 16S rDNA. Spectrum of the antibacterial agent was also tested against routine observed bacteria with drinking water contamination such as Escherichia coli, Salmonella, Pseudomonas, and Vibrio and these were found to be sensitive. Bacteria like Acinetobacter and Burkholderia were found to be resistant. The differential antibacterial activity of the HPC-250 was observed for the genus Bacillus where B. subtilis remained resistant although B. sphaericus was sensitive. PMID- 16211432 TI - Purification and characterization of two novel antimicrobial peptides Subpeptin JM4-A and Subpeptin JM4-B produced by Bacillus subtilis JM4. AB - An antimicrobial peptides-producing strain was isolated from soil and identified as Bacillus subtilis JM4 according to biochemical tests and 16S rDNA sequence analysis. The corresponding antimicrobial peptides were purified to homogeneity by ammonium sulfate precipitation, sequential SP-Sepharose Fast Flow, Sephadex G 25 and C18 reverse-phase chromatography, and in the final purification step, two active fractions were harvested, designated as Subpeptin JM4-A and Subpeptin JM4 B. The molecular weights, determined by mass spectrometry, were 1422.71 Da for Subpeptin JM4-A and 1422.65 Da for Subpeptin JM4-B, respectively. Amino acid sequencing showed that they differed from each other only at the seventh amino acid except for three unidentified residues, and the two peptides had no significant sequence homology to the known peptides in the database, indicating that they are two novel antimicrobial peptides. In addition, characteristic measurements indicated that both peptides had a relatively broad inhibitory spectrum and remained active over a wide pH and temperature range. PMID- 16211433 TI - Rhizosphere competent Pseudomonas aeruginosa GRC1 produces characteristic siderophore and enhances growth of Indian mustard (Brassica campestris). AB - Pseudomonas aeruginosa GRC1, an isolate of potato rhizosphere, was known to have several plant growth-promoting activities, including production of phytohormone and antibiotic substance. The isolate was found to have prolific production ability of hydroxamate siderophore in iron-deficient conditions. The siderophore of GRC1 was purified and characterized. The purified siderophore appeared to be of pyoverdin type with typical amino acid composition. In field trials, P. aeruginosa GRC1 enhanced the growth of Brassica campestris var Pusa Gold (Indian mustard). Significant increase in root and shoot weight, length, grain yield per plant, and total grain yield was recorded. Root colonization was studied with Tn5 induced streptomycin-resistant transconjugants of spontaneous rifampicin resistant GRC1 (designated GRC1(rif+strep+)) after different durations. The strain was significantly rhizopheric competent and stabilized in the rhizosphere, without disturbing the normal indigenous bacterial population. PMID- 16211435 TI - Pneumoperitoneum does not influence trocar site implantation during tumor manipulation in a solid tumor model. AB - BACKGROUND: The purpose of this study was to assess tumor implantation at abdominal wound sites following manipulation of a solid abdominal tumor. METHODS: GW-39 human colon cancer cells were injected into the omentum of golden Syrian hamsters. At 2 weeks, an omental tumor was harvested and animals were randomized to bivalve (A), crush (B), strip (C), or excision (D), with or without pneumoperitoneum. Four 5-mm trocars were inserted into the abdomen, and the tumor was reinserted through the midline, swept through four quadrants, and removed. The incision was closed and pneumoperitoneum at 7 mmHg was maintained for 10 min. Tumor implantation at wound sites was documented at 7 weeks. RESULTS: Implantation at trocar sites was 53 and 49% with and without pneumoperitoneum in the manipulated groups (A, B, C), respectively (p = 0.993). Implantation at trocar sites was reduced in the control group (D) at 9 and 10% with and without pneumoperitoneum, respectively (p < 0.001). CONCLUSIONS: Tumor implantation at trocar sites is due to spillage of tumor during manipulation and not to pneumoperitoneum. PMID- 16211434 TI - Molecular cloning and genetic analysis of functional merB gene from indian isolates of Escherichia coli. AB - Studies were carried out to characterize organomercurial lyase genes from wild type mercury-resistant Escherichia coli isolates, previously collected from five geographically distinct regions of the Indian subcontinent. PCR amplification followed by DNA sequencing of amplified fragments showed three merB identical to the previously characterized mer B from E. coli pR831b that were thus considered as the same gene. The remaining two genes derived from E. coli isolates of an almost mercury-free site (Dal lake, Kashmir) and designated as pIAAD3 merB and pIAAD14 merB showed slight variation (2%) at base. However, this variation in pIAAD3 due to the absence of base "T" at 479 position results in complete frame shift and the predicted MerB-like polypeptide derived from it showed 21.53% divergent at its C terminal end from the previously characterized pR831b MerB. The expression profile of pIAAD3 merB in pQE30 and pUC18 vectors each demonstrated 22.2 kDa proteins. The induced DH5alpha E. coli cells possessing pIAAD3 merB cloned in pUC18 vector split phenyl mercuric acetate (PMA) into benzene and inorganic mercury efficiently, thus giving a clue that the expressed gene product is biologically active. The current study suggests that such genetic changes may take place in the continued absence of mercury pressure, and with such modifications, they finally break down to act as vestigial remnants. Further work is going on in our lab to exploit pIAAD3 merB for the bioremediation of mercury-polluted sites. PMID- 16211436 TI - Endoscopic ultrasound for preoperative staging of esophageal carcinoma. AB - BACKGROUND: Endoscopic ultrasound (EUS) is potentially the best method for pretreatment staging of esophageal carcinoma once distant metastases have been excluded by other methods. However, its apparent accuracy might be influenced by the use of neoadjuvant therapy. To determine the accuracy of EUS in patients undergoing esophageal resection, the authors reviewed their experience with EUS. METHODS: A total of 73 patients with esophageal carcinoma who underwent an esophagectomy between April 2000 and February 2005 were examined using preoperative EUS and computed tomography (CT). Of these patients, 39 also underwent preoperative neoadjuvant chemoradiotherapy. Both EUS and CT scan were used to determine the depth of tumor penetration (T-stage) and the presence of lymph node metastases (N-stage). These results then were compared with staging determined after pathologic examination of the resected surgical specimen. RESULTS: For patients not undergoing neoadjuvant therapy, T-stage was accurately determined by EUS in 79%, N-stage in 74%, and tumor node metastasis (TNM) classification in 65% of the cases. However, when patients who had undergone neoadjuvant chemoradiotherapy were included, the overall accuracy of EUS was 64% for T-stage, 63% for N-stage, and 53% for TNM classification. For the patients who underwent neoadjuvant therapy, EUS indicated a more advanced T-stage in 49%, N-stage in 38%, and TNM classification in 51% of the cases, as compared with pathology. The overall accuracy of EUS for T- and N-stage carcinomas was superior to that of CT scanning. CONCLUSION: For patients who do not undergo preoperative neoadjuvant chemotherapy and radiotherapy, EUS is a more accurate method for determining T- and N-stage resected esophageal carcinomas. Neoadjuvant therapy, however, results in apparent overstaging, predominantly because of tumor downstaging, and this reduces the apparent accuracy of EUS (and CT scanning) in this patient group. Nevertheless, EUS staging before neoadjuvant therapy could be more accurate than pathologic staging after treatment, thereby providing better initial staging information, which can be used to facilitate treatment. PMID- 16211437 TI - Gastric carcinoids: a temporal increase with proton pump introduction. AB - BACKGROUND: Recent reports have indicated a rising incidence of gastric carcinoids. This study aimed to evaluate the incidence pattern of gastric carcinoids in two large population-based cancer registries. METHODS: The Florida Cancer Data System (FCDS), Florida's statewide cancer registry, and the Surveillance, Epidemiology, and End Results (SEER) program were used. The study population was defined as all cases of gastric carcinoid identified in either database from January 1981 to December 2000. Descriptive statistics and age adjusted incidence rates were calculated. RESULTS: There were 326 (FCDS) and 594 (SEER) cases of invasive gastric carcinoid during the 20-year study period. The mean age of the patients was 65 years (range, 21-96 years), and the male:female ratio was 1:1. The age-adjusted incidence rate in FCDS increased from 0.04 (per 100,000 age-adjusted to the 2000 U.S. standard population) to 0.18 in the year 2000. The estimated annual percentage change in incidence was 8.17 in FCDS and 9.17 in SEER (p < 0.05). A decrease in gastric cancer was noted during this same period (from 8.64 to 11.14 cases per 100,000 in FCDS and from 11.14 to 8.06 cases per 100,000 in SEER). CONCLUSIONS: This study documented a statistically significant eight- or ninefold increase in the incidence of gastric carcinoids in two large databases. The temporal increase in incidence correlates with the introduction and widespread use of proton pump inhibitors since the late 1980s. Other explanations include improved detection with wider application of upper endoscopy. Further epidemiologic studies are warranted. PMID- 16211438 TI - Techniques and results of laparoscopic antireflux surgery in Germany. AB - BACKGROUND: This study aimed to evaluate the development and outcomes of laparoscopic antireflux surgery in Germany using a nationwide representative survey. METHODS: A written questionnaire including 34 detailed questions and 288 structured items about diagnostic and therapeutic approaches, number of procedures, complications, and mortality was sent to 546 randomly selected German surgeons (33% of the registered general surgeons) at the end of 2000. RESULTS: The response rate was 72%, and a total of 2,540 antireflux procedures were reported. According to the survey, 81% of all procedures were performed laparoscopically, and 0.1% were performed thoracoscopically. As reported, 65% were total fundoplications, 31% were partial fundoplications, and 4% were other procedures. Of the surgeons who had experience with laparoscopic antireflux techniques (29%), 71% preferred a 5-trocar technique, and 91% used the Harmonic Scalpel for dissection. There were significant technical variations among the surgical procedures (e.g., use and size of the bougie, length of the wrap, additional gastropexy, fixation of the wrap). The overall complication rate for laparoscopic fundoplication was 7.7% (5.7% surgical and 2% nonsurgical complications), including rates of 0.6% for esophageal perforations and 0.6% for splenic lesions. The conversion rate was 2.9%; the reoperation rate was 1.6%; and the overall hospital mortality rate was 0.13%. The authors observed a striking learning curve difference in complication rates between hospitals performing fewer than 10 laparoscopic antireflux techniques annually and those performing more than 10 fundoplications per year (14% vs 5.1%, p < 0.001). Long-term dysphagia and interventions occasioned by dysphagia occurred significantly more often after total fundoplications than after partial fundoplications (6.6% vs 2.4%; p < 0.001). Similar findings were reported for Nissen versus floppy Nissen procedures. The overall failure rate, however, was similar for both groups (Nissen 8.7%; partial 9%, difference not significant). CONCLUSIONS: Until now, no unique laparoscopic antireflux technique has been accepted, and a number of different antireflux procedures with numerous modifications have been reported. The morbidity and mortality rates reported in this article compare very well with those in the literature, and 1-year-follow-up results are promising. PMID- 16211439 TI - Comparative anatomy of the porcine and human thoracic spines with reference to thoracoscopic surgical techniques. AB - BACKGROUND: This study compared porcine and human thoracic spine anatomies for a better understanding of how structures encountered during thoracoscopy differ between training with a porcine model and actual surgery in humans. METHODS: Parameters were measured including vertebral body height, width, and depth; disc height; rib spacing; spinal canal depth and width; and pedicle height and width. RESULTS: Although most porcine vertebral structures were smaller, porcine pedicle height was significantly greater than that of humans because the porcine pedicle houses a unique transverse foramen. The longus colli and psoas attach, respectively, to T5 and T13 in swine and to T3 and T12 in humans. In swine, the azygos vein generally was absent. The intercostal veins drained into the hemiazygos vein. CONCLUSIONS: Several thoracoscopically relevant anatomic differences between human and porcine spinal anatomies were identified. A thoracoscopic approach in a porcine model probably is best performed from the right side. The best general working area is between T6 and T10. PMID- 16211441 TI - Major mesh-related complications following hernia repair: events reported to the Food and Drug Administration. AB - Mesh material affects complications following hernia repair. Medical device reports on the use of surgical mesh for hernia repair were reviewed from the Food and Drug Administration's (FDA) Manufacturer User Facility Device Experience Database from January 1996 to September 2004. We analyzed 252 adverse event reports related to the use of surgical mesh for hernia repair. Adverse events included infection (42%, 107 reports), mechanical failure (18%, 46), pain (9%, 23), reaction (8%, 20), intestinal complications (7%, 18), adhesions (6%, 14), seroma (4%, 9), erosion (2%, 6), and other (4%, 9). Compared to all other mesh types, Sepra/polypropylene mesh had more mechanical failures (80 vs 14%, p < 0.05), biomaterial mesh had more reactions (57 vs 7%, p < 0.05), polytetrafluoroethylene (PTFE)/polypropylene mesh had more intestinal complications (14 vs 7%, p < 0.05), and PTFE mesh tended to have more infections (75 vs 41% all other, p = 0.07). Death occurred in 2% (5). We conclude that specific mesh materials are related to specific complications. PMID- 16211440 TI - Comparison of laparoscopic versus open repair for perforated duodenal ulcers. AB - BACKGROUND: There is great controversy regarding the choice of procedure for perforated duodenal ulcer patients. The purpose of this study was to compare the early outcome results of laparoscopic and open repair and to propose which risk factors influence the outcome. METHODS: Between October 1996 and May 2004, 60 patients underwent laparoscopic and 162 patients underwent open repair of perforated peptic ulcers in a tertiary care academic center. The results were retrospectively analyzed. The primary outcome measures included operative time, duration of hospital stay, morbidity, and mortality. RESULTS: The operative time was significantly longer in the laparoscopy group compared to the open repair group (76.2 +/- 35.3 vs 57.3 +/- 26.1 min, respectively). The hospital stay in surviving patients appeared to be significantly shorter after laparoscopy than after open repair (7.8 +/- 5.3 vs 10.3 +/- 10.6 days, respectively). Eight patients (13%) in the laparoscopic group and 41 patients (25%) in the open repair group had morbidity in the postoperative period. Suture leakage was confirmed in four patients (7%) following laparoscopic repair and in three patients (2%) in the open repair group. There were 20 deaths (9%), all in the open repair group. CONCLUSIONS: Independent Boey risk factors, patient age, and large perforation size have a negative impact on patient recovery. Both laparoscopic and open repair are equally safe and effective in perforated duodenal ulcer patients with a Boey score of 0 or 1. PMID- 16211442 TI - Diamniotic conjoined fetuses in a triplet pregnancy: an insight into embryonic topology. AB - We present a case of triplets, 2 of whom were monochorionic diamniotic conjoined fetuses, and the other triplet was in a separate chorion. The pregnancy followed in vitro fertilization with 2 embryo transfers and the conjoined fetuses developed from a single embryo. An early ultrasound showed 2 embryos attached to 1 yolk sac. Further monitoring of the pregnancy showed these 2 triplets to be diamniotic with a short umbilical cord/body stalk anomaly. After fetal loss a postmortem examination demonstrated that these fetuses were conjoined with a body stalk anomaly/short umbilical cord syndrome with fusion of the coelomic cavities and small bowel (minimally united/conjoined twins). There were dividing amniotic membranes. This case demonstrates the development of an unusual type of conjoined twin with characteristic features. Conjoined fetuses with this pattern have fused small bowel and cloacal anomalies and often diamniotic placentation, when this is recorded. Various terms including minimally united/conjoined omphalopagus/ischiopagus have been used for this characteristic pattern, but this pattern may warrant a specific term. "Diamniotic vitellopagus" may best reflect the pathogenesis of this pattern. PMID- 16211443 TI - Extranodal Rosai-Dorfman disease with multifocal bone and epidural involvement causing recurrent spinal cord compression. AB - Sinus histocytosis with massive lymphadenopathy, or Rosai-Dorfman disease, is a rare histiocytic disorder that typically presents with chronic, self-limiting, cervical lymphadenopathy. We present a case, a diagnostic dilemma for multiple consultation services, of an otherwise well 17-year-old boy without lymphadenopathy who, 8 months after excision of a T9 lytic vertebral lesion and epidural mass that caused cord compression, again presented with cord compression from progressive vertebral disease, recurrent epidural mass, and development of a paraspinal mass and tibial lesion. The excised vertebral and epidural lesions, 2 paraspinal biopsies, and tibial biopsy were interpreted as chronic inflammation until large histiocytes were noted, which were positive for CD68, S100 protein, fascin, and MAC387, and demonstrated characteristic emperipolesis (lymphophagocytosis) that was diagnostic of Rosai-Dorfman disease. This atypical clinical behavior and sites of involvement of multiple bones but not of lymph nodes is unusual and constitutes the aggressive end of the clinical spectrum and a rare cause for cord compression. PMID- 16211444 TI - Adenomyomatosis of the gallbladder in childhood: report of a case and review of the literature. AB - Adenomyomatosis of the gallbladder is considered an acquired pathologic condition that affects adults, with only 2 cases previously described in childhood. We report on a 5-year-old boy with incidental detection of this lesion by ultrasonography. Pathologic and clinical assessments are discussed in addition to treatment decision-making, including current insights into the relation between adenomyomatosis and malignancy of the gallbladder. The liberal use of ultrasonography in children with abdominal pain may result in detection of an increasing number of asymptomatic patients. Caregivers should be aware of this condition and its therapeutic implications in the pediatric population. PMID- 16211445 TI - Temporal soft tissue glioneuronal heterotopia in a child with a seizure disorder: case report and review of the literature. AB - Tissue and cellular elements generally attributed to the central nervous system (CNS) are infrequently found in areas outside the CNS proper. Most of these lesions contain predominantly glial tissue. In rare instances, heterotopic CNS tissue is found in the scalp, many associated with an intracranial connection and overlying skin and hair anomalies. In follow-up of these patients, development is normal. We present a case of a temporal scalp mass in a 19-month-old girl with a seizure disorder beginning at about age 1 year. At the time of excision, the mass was not associated with intracranial connection or overlying cutaneous abnormalities and demonstrated abundant neuronal and glial elements with features of dysplastic CNS tissue. We speculate that, in certain circumstances, aberrantly localized CNS tissue may be related to abnormal CNS development. PMID- 16211446 TI - Partly molar pregnancies that are not partial moles: additional possibilities and implications. PMID- 16211447 TI - Cytogenetic findings in pediatric T-lymphoblastic lymphomas: one institution's experience and a review of the literature. AB - Cytogenetic analyses of lymphomas commonly reveal nonrandom chromosomal abnormalities, but there are relatively few reports in childhood lymphoblastic lymphoma (LL). We retrospectively reviewed G-banded karyotypic analyses performed at Arkansas Children's Hospital between 1990 and 2004. Six children (2 to 20 years old) had LL that presented as mediastinal or cervical masses and had a T cell immunophenotype and clonal abnormalities. The cytogenetic findings in these 6 patients were as follows: 46,XX,-7,inv(9)(p11q12),der (12)t(7;12)(q11.2;p13),t(16;18)(p13.1;q21),+22 in patient 1; 47,XX,+9,del(9)(q11q22)x2 in patient 2; 72-119, XY,+X,+1,+1, inv(2) (p11q13), 3,+5,+6,+7,+10,-12,-16, -21,-21,-22,+mar in patient 3; 48,XY,+5,+20,t(7;9) (q32;q34) in patient 4; 47 approximately 48,XX,der(10)t(10;14)(q23; q11.2),+12, del(12)(p12)x2, -14,del(16)(q22q22),+?add (19)(p13.3) in patient 5; and 48 approximately 49,XY,+7,+8,t(11;19) (q23;p?13.3),+der(19)t(11;19)[cp20] in patient 6. Eleven chromosome breakpoints in 6 of our patients (7q11, 12p13, 16p13, 18q21, 9q11, 2p11, 2q13, 7q32, and 7q23) have been reported in other patients with acute lymphoblastic leukemia or LL and involved regions containing TEL, ABL, E2A, MLL, and T-cell receptor-alpha genes. A review of the cytogenetic findings of these and other cases of LL reveals that clonal aberrations are common and most frequently involve T-cell receptor gene regions. The aberrations show some features similar to those of acute lymphoblastic leukemia and are not unique to LL, thus furnishing additional evidence of the equivalence of these two diseases. The cytogenetic features of LL may be helpful in the diagnosis of pediatric lymphomas and undifferentiated neoplasms. PMID- 16211448 TI - HER2 amplification and overexpression is not present in pediatric osteosarcoma: a tissue microarray study. AB - The HER2 gene, located on 17q, encodes a 185-kD transmembrane tyrosine kinase receptor. Amplification of this gene with overexpression of the gene product occurs in about 30% of cases of breast cancer and is considered to be a poor prognostic indicator for this tumor. Results for HER2 expression in osteosarcoma are controversial, with some studies reporting up to 61% of positive cases and others reporting only negative results. Further, expression of HER2 is reported to be a favorable prognostic indicator by some groups and unfavorable by others. The present study used tissue microarrays containing 34 samples of osteosarcoma from 18 patients to analyze HER2 expression by immunohistochemistry and gene copy number by chromogenic in situ hybridization. The microarray included 13 pretreatment biopsies, 11 posttreatment resection specimens, and 10 resected metastases and comprised 18 osteoblastic, 6 chondroblastic, 5 fibroblastic, and 5 mixed subtypes. HER2 protein expression was seen in 4 of 34 (12%) tumor samples that originated from 2 of 18 patients (11%). The staining pattern was consistently weak and focal, and immunohistochemical overexpression of the HER2 protein, defined as complete membrane positivity, was never observed. Further, the presence of HER2 gene amplification was not detected in any osteosarcoma by chromogenic in situ hybridization. Therefore, therapies based on antibodies directed against the HER2 protein are unlikely to have much value in the treatment of pediatric osteosarcomas. From a technical standpoint, this study also demonstrates the value of tissue microarrays in screening tumors at the protein and gene levels using conventional light microscopy. PMID- 16211449 TI - Review of tonsillar lymphoma in pediatric patients from the pediatric oncology group: what can be learned about some indications for microscopic examination? AB - Financial considerations have led to suggestions that routine microscopic evaluation of tonsils and adenoids is neither cost effective nor clinically indicated. However, the possibility of tonsillar lymphoma must be carefully weighed when making institutional policy decisions. One way to find an appropriate algorithm for pathologic examination is to examine the characteristics of biopsy-proved tonsillar lymphomas. To investigate the clinicopathologic characteristics of tonsillar lymphoma, we performed a retrospective analysis of patients who had non-Hodgkin lymphoma (NHL) and large cell or Burkitt lymphoma involving the tonsils and adenoids and were registered on Pediatric Oncology Group (POG) protocols. Seventy-seven (9%) of 832 POG cases of NHL involved the tonsils and adenoids. Review of the pathology reports available from 29 of these cases revealed that NHL was incidentally discovered by pathologic examination in only 5 cases, all of which had clinical evidence of unilateral tonsillar enlargement or size discrepancy by gross examination. The other 24 cases indicated a clinical suspicion of tonsillar cancer, as judged by a preoperative diagnosis or by a request for frozen-section examination. We conclude that in most cases there is a clinical suspicion of tonsillar NHL at the time of gross examination. With routine cases, we predict that the use of comparative organ weights, a clinical impression of tonsillar asymmetry, and review of clinical history will increase the recognition of tonsillar lymphoma when "gross-only" protocols are employed for specimen handling. PMID- 16211450 TI - Sudden death in a neonate with idiopathic eosinophilic endomyocarditis. AB - A 26-day-old male infant who had been fussy and feeding poorly for a period of several hours died suddenly despite efforts at resuscitation. Postmortem examination revealed eosinophilic endomyocarditis unassociated with disease in other organs. The etiology remained unexplained after review of the medical and family histories and circumstances of death, extensive light and immunofluorescence microscopies, and microbiological, metabolic, and toxicologic testing. This appears to be the youngest reported case of eosinophilic endomyocarditis, a disorder that typically occurs in middle-age males and is often associated with benign or malignant hypereosinophilic conditions. The medical literature is reviewed. PMID- 16211451 TI - Telangiectatic focal nodular hyperplasia of the liver in the perinatal period: case report. AB - We report a case of congenital telangiectatic focal nodular hyperplasia, a rare variant form of the disease. The patient was a 2-month-old boy whose parents noticed abdominal distention about 2 weeks after birth, and ultrasonogram revealed a large mass in the liver. He underwent right lobectomy, and gross findings showed an ill-defined mass without any central scar. Histologic findings demonstrated proliferating hepatocytes without atypia arranged in cords of 1- or 2-cell thickness with marked sinusoidal dilatation and extramedullary hematopoiesis. In addition, a significantly increased Ki-67 labeling index in the tumor compared with non-tumor liver cells, and cytogenetic analysis of 23 G banded metaphase preparations revealed 3 abnormal karyotypes, suggesting hyperplastic or neoplastic features. To the best of our knowledge, the present case is only the third documented case of congenital telangiectatic focal nodular hyperplasia. PMID- 16211452 TI - Pulmonary interstitial hemosiderin in infancy: a common consequence of normal labor. AB - Deposits of hemosiderin are found frequently in the interstitium of the lungs of infants who die suddenly and have been a suggested marker for hypoxia during previous episodes of apnea. We studied the epidemiology of pulmonary interstitial hemosiderin (PIH) in 94 infants with a diagnosis of the sudden infant death syndrome that was established during the New Zealand Cot Death Study. Twenty seven infants (29%) had widely distributed PIH. Associations were sought between PIH and variables on which information had been obtained from parental interviews or from medical records. Previous suggestions of associations with increasing birth weight and decreasing age at death were confirmed in multivariate analysis (P = 0.0018 and P = 0.03, respectively) and with increasing gestation in univariate analysis (P < 0.0001). PIH was not found in any of the 13 infants delivered by elective cesarean section compared with a 33% incidence in those exposed to labor (P = 0.02). There was no association with previous apnea. We consider that the likely explanation of these findings is that PIH is a residue of interstitial hemorrhage caused by chest compression during labor that occurs particularly in larger infants of greater gestational age and then clears gradually during early infancy. PMID- 16211453 TI - Light microscopic, immunohistochemical, and ultrastructural findings in congenital fibular aplasia or hypoplasia (FAH). AB - Congenital aplasia or hypoplasia of the fibula (FAH) is a rare malformation that is defined by a partial or complete absence of the fibular bone. Etiology and pathogenesis are unknown and the precise morphology of the tissue cord replacing the malformed fibula has not been well described. Therefore, tissue cord was examined in 8 patients with FAH. Light microscopic, immunohistochemical, and electron microscopic investigations showed a core of embryonic cartilage with collagen II and VI expressions surrounded by connective tissue. Although collagen II expression is typical for chondroid differentiation, collagen VI reactivity is normally seen in articular cartilage and tendon-like fibrocartilaginous tissue but is absent in hyaline cartilage. Further ultrastructural analyses by electron microscopy supported these findings. The histomorphologic changes correspond to the histologic findings of Papenbrock et al. (2000, Mech Dev 92:113-123) who produced a congenital malformation in transgenic mice that resembled FAH by overexpression of Hox c11. PMID- 16211454 TI - beta-Catenin and met deregulation in childhood Hepatoblastomas. AB - Activation of the Wnt/beta-catenin and hepatocyte growth factor/Met signaling has been implicated in various tumors. Owing to the cross-talk between these pathways and aberrant redistribution of beta-catenin in hepatoblastomas, we examined their status in this tumor. This study examined changes in beta-catenin and Met in paired pretreatment and post-treatment hepatoblastoma tissues in relation to their effects on proliferation and target genes such as c-myc and cyclin-D1. In this study we compared proliferation indices, beta-catenin staining and its known molecular targets, c-myc and cyclin-D1, and Met, a tyrosine kinase receptor for hepatocyte growth factor in pretreatment and post-treatment specimens. Pretreatment and post-treatment sections from 13 children, ages 11 weeks to 9 years, were analyzed for these markers by immunohistochemistry. All tumors (13 of 13) displayed increased proliferation and beta-catenin (cytoplasmic and nuclear) staining in pretreatment biopsies that remained relatively unaffected after treatment. Aberrant Met staining (cytoplasmic) was observed in all pretreatment samples that decreased considerably after treatment in 11 of 13 patients. A significant subset of these tumors showed increased c-myc and cyclin-D1 staining in pretreatment biopsies that decreased after chemotherapy in most cases. beta Catenin redistribution in tumor cells corresponds to proliferation in hepatoblastomas. However, beta-catenin nuclear localization remains unaffected in viable hepatoblastoma tissue after chemotherapy. In contrast, Met undergoes a prominent decrease after treatment and thus might be important in pathogenesis of hepatoblastoma. PMID- 16211455 TI - Multinucleated foreign body giant cells in placental membrane. AB - We report the pathologic finding of a multinucleated foreign body giant cell reaction to squames and fetal hair in the placental membranes in a 37-week 1-day intrauterine gestation. This reaction appeared to have developed in association with repeated intrauterine procedures performed in the third trimester, including cordocentesis for fetal blood sampling, intrauterine blood transfusion, and amnioreduction for polyhydramnios. This type of reaction most likely was directed to prolonged amniotic fluid leakage that occurred spontaneously or after intrauterine procedures in the second half of the second trimester and the third trimester. Careful examination of the placental membranes and recognition of the foreign body giant cell reaction may provide etiologic insight in cases of unexplained oligohydramnios. PMID- 16211456 TI - Lymphatics in the alimentary tract of children in health and disease: study on mucosal biopsies using the monoclonal antibody d2-40. AB - Primary intestinal lymphangiectasia and intestinal lymphatic hypoplasia are 2 causes of protein-losing enteropathy in children and share many common clinical features. For the diagnosis of lymphatic hypoplasia on endoscopic biopsies of the intestine, i.e., based on a negative finding in a small specimen, a very sensitive and specific method for identifying lymphatics is essential. In the present study, lymphatic vessels were labelled using D2-40 immunostaining in mucosal biopsy specimens of the alimentary tract of children in whom no histologic abnormality was noted and of those who had different relatively common pediatric conditions, including inflammatory and neoplastic diseases. Using this method, lymphatic vessels were well visualized even in young infants and not destroyed by diseases. The presence of the muscularis mucosae in specimens was important for adequate assessment. In the duodenum and esophagus, lymphatics were observed in every single section; in the stomach, ileum, and colon, they were less regular and several sections were sometimes required. The extreme sensitivity of this method for demonstrating lymphatic vessels in the duodenum makes it ideal for the histologic diagnosis of intestinal lymphatic hypoplasia. In 4 patients who were considered to have this diagnosis based on clinical features, full-thickness intestinal biopsies and electron microscopy, D2-40 immunostaining confirmed the absence or marked paucity of lymphatics. PMID- 16211457 TI - Tuboovarian abscess in a 7-year-old girl with markedly increased tumor markers in the absence of malignancy. PMID- 16211458 TI - Three-dimensional reconstruction and morphologic measurements of human embryonic hearts: a new diagnostic and quantitative method applicable to fetuses younger than 13 weeks of gestation. AB - Improvements in the diagnosis of congenital malformations explain the increasing early termination of pregnancies. Before 13 weeks of gestation, an accurate in vivo anatomic diagnosis cannot currently be made in all fetuses with current imaging instrumentation. Anatomopathologic examinations remain the gold standard to make accurate diagnoses, although they reach limits between 9 and 13 weeks of gestation. We present the first results of a methodology that can be applied routinely, using standard histologic section, thus enabling the reconstruction, visual estimate, and quantitative analysis of 13-week human embryonic cardiac structures. The cardiac blocks were fixed, embedded in paraffin, and entirely sliced by a microtome. One of 10 slices was topographically colored and digitized on an optical microscope. Cardiac volume was recovered by semiautomatic realignment of the sections. Another semiautomatic procedure allowed extracting and labeling of cardiac structures from the volume. Structures were studied with display tools, which disclosed the internal and external cardiac components and enabled determination of size, thickness, and precise positioning of ventricles, atria, and large vessels. This pilot study confirmed that a new 3-dimensional reconstruction and visualization method enables accurate diagnoses, including in embryos younger than 13 weeks. Its implementation at earlier stages of embryogenesis will provide a clearer view of cardiac development. PMID- 16211459 TI - Different effects of growth factors on proliferation and matrix production of normal and fibrotic human lung fibroblasts. AB - OBJECTIVES AND METHODS: In idiopathic pulmonary fibrosis (IPF), proliferation of fibroblasts and increased matrix deposition result in pulmonary damage and respiratory insufficiency. We cultured human fibroblasts from lung biopsies of healthy adults and of three patients with IPF (histologically usual interstital pneumonitis, UIP) in order to compare proliferation ([(3)H]thymidine incorporation, cell count) and matrix protein expression (immune fluorescence, quantification of fibronectin synthesis using time-resolved immune fluorescence) of normal and UIP fibroblasts in response to various growth factors. FINDINGS: The growth factors platelet-derived growth factor-BB (PDGF), epidermal growth factor (EGF), insulin growth factor-1 (IGF-1), insulin-like growth factor-2 (IGF 2), tumor necrosis factor alpha (TNFalpha), Transforming growth factor-beta (TGFbeta(1)), and fibroblast growth factor-2 (FGF-2) stimulate proliferation of normal lung fibroblasts significantly more than proliferation of UIP fibroblasts. Immunofluorescence reveals extensive expression of collagen I, collagen III, and fibronectin induced by serum, TGFbeta(1), and TNFalpha. This expression is more pronounced in UIP fibroblasts than in normal fibroblasts. Quantification of fibronectin synthesis reveals an enhanced fibronectin synthesis by UIP fibroblasts in response to PDGF, EGF, IGF-1, IGF-2, TNFalpha, TGFbeta(1), and FGF 2). CONCLUSIONS: Fibroblasts from normal and UIP lungs differ in their response to growth factors: Whereas normal fibroblasts show a predominantly proliferative response, UIP fibroblasts show an enhanced synthetic activity. Different fibroblast responses may contribute to progressive pulmonary fibrosis in patients with UIP. PMID- 16211460 TI - Gammaherpesvirus-induced lung pathology is altered in the absence of macrophages. AB - The purpose of this study was to examine the lung pathogenesis of murine gammaherpesvirus (MHV-68) infection in mice that lack CC chemokine receptor CCR2, an important receptor for macrophage recruitment to sites of inflammation. BALB/c and CCR2(-/-) mice were inoculated intranasally (i.n.) with MHV-68 and samples were collected during acute infection (6 dpi) and following viral clearance (12 dpi). Immunohistochemistry was used to determine which cells types responded to MHV-68 infection in the lungs. Lung pathology in infected BALB/c mice was characterized by a mixed inflammatory cell infiltrate, necrosis, and increased alveolar macrophages by 12 dpi. Immunohistochemistry showed intense positive staining for macrophages. CCR2(-/-) mice showed greater inflammation in the lungs at 12 dpi than did BALB/c mice, with more necrosis and diffuse neutrophil infiltrates in the alveoli. Immunohistochemistry demonstrated much less macrophage infiltration in the CCR2(-/-) mice than in the BALB/c mice. These studies show that CCR2 is involved in macrophage recruitment in response to MHV 68 infection and illustrates how impairments in macrophage function affect the normal inflammatory response to this viral infection. PMID- 16211461 TI - Influence of mast cells on the expression of adhesion molecules on circulating and migrating leukocytes in acute pancreatitis-associated lung injury. AB - Pancreatitis-associated lung injury is an early-occurring and severe complication, still associated with substantial mortality. A number of inflammatory cells and their products are involved in the initiation and progress of the condition. In the present study, acute pancreatitis (AP) was induced by the intraductal infusion of 5% sodium taurodeoxycholate in the rat. Pulmonary endothelial barrier dysfunction was measured by plasma exudation of radiolabeled albumin. Expression of PECAM-1, ICAM-1, and L: -selectin on neutrophils (CD11b(+)) and monocytes/macrophages (CD11b/c(+)), obtained from circulation and lung tissue, was measured 1 and 6 hours after AP induction (n = 10 rats/time point/group). Plasma levels of histamine and serotonin were determined. The role of mast cells was evaluated by pretreatment with the mast cell stabilizer cromolyn. Intraperitoneal administration of cromolyn downregulated pancreatitis induced systemic increase of histamine at 1 hour (513 +/- 82 vs. 309 +/- 50, p < 0.05). Cromolyn prevented a decreased expression of PECAM-1 on circulatory neutrophils and monocytes/macrophages and against an increased expression of ICAM 1 and PECAM-1 on pulmonary neutrophils and monocytes/macrophages 6 hours after AP induction (about 40% vs. 10%, p < 0.01). The mast cell stabilizer also prevented pancreatitis-induced pulmonary endothelial barrier dysfunction at 6 hours. Thus, our data indicate that mast cells may play a critical role in the activation of leukocytes during the initiation of pancreatitis-associated lung injury by altering phenotypes of adhesion molecules. PMID- 16211462 TI - Phase II study of troxacitabine (BCH-4556) in patients with advanced non-small cell lung cancer. AB - Troxacitabine. a promising new L-nucleoside, inhibits DNA polymerase and leads to complete DNA chain termination. The National Cancer Institute of Canada Clinical Trials Group (NCIC-CTG) conducted a phase II study to assess the efficacy and toxicity of troxacitabine in untreated patients with advanced non-small-cell lung cancer (NSCLC). Previously untreated patients were eligible if they had inoperable stage IIIB or IV NSCLC, ECOG PS < or = 2, adequate hematology and biochemistry, and at least one bidimensionally measurable lesion. Patients with prior malignancy or brain metastases were excluded. Troxacitabine (10 mg/m(2)) was administered intravenously over 30 minutes every 3 weeks. Between June 1999 and May 2000, 17 eligible patients received treatment. Patient characteristics included: median age 64 years; female 41%; stage IV (94%); PS 0 (12%), 1 (59%), and 2 (29 %), 3 or more disease sites (59%). In 17 patients, there were 8 stable disease, 9 disease progression, and no objective responses. Median duration of stable disease was 3.6 months (range = 2.0-7.1). A total of 56 cycles were administered (median = 3), and 88% of patients received 90% or more of the planned dose intensity. The majority (82%) of patients experienced skin rash. Hematologic and biochemical toxicities, grade 3/4 (%) were: granulocytopenia (41), anemia (12), thrombocytopenia (6), and hyperglycemia (6). Troxacitabine appears to have little activity in NSCLC in the dose and schedule tested. PMID- 16211463 TI - Development of microscopic polyangiitis in patients with chronic airway disease. AB - Microscopic polyangiitis (MPA) is a rare systemic vasculitis syndrome, which is often accompanied by positive myeloperoxidase-specific antineutrophil cytoplasmic antibody (MPO-ANCA). While pulmonary involvement of MPA consists mainly of diffuse alveolar hemorrhage and interstitial pneumonia, bronchiectasis has been reported as a pulmonary lesion in association with MPA. To investigate the clinical features of patients with MPA, focusing on the presence or the absence of preceding chronic airway diseases (CAD), we conducted a retrospective observational study of 26 patients in the last 13 years at Saga University Hospital. The clinical records and radiologic chest examinations were reviewed retrospectively. Pulmonary manifestations were alveolar hemorrhage in 3 patients (12%) and interstitial pneumonia in 5 (19%). Bronchiectasis, defined by the findings of chest radiograph and computed tomography, was found in 9 patients (35%). Four patients (15%) with bronchiectasis and one patient (4%) with chronic bronchitis had experienced chronic bronchial suppuration prior to the onset of MPA. Ten patients were classified as having chronic airway disease (CAD) before the onset of MPA. MPO-ANCA tended to be lower in the CAD group than in the non CAD group. None of the patients in the CAD group had pulmonary hemorrhage or interstitial pneumonia. Only one patient (10%) in the CAD group died within 90 days of the onset of MPA, while 7 (43.8%) of the non-CAD group died. Our study suggests that MPA may result in part from CAD and that the clinical course of MPA with CAD may be different from MPA without CAD. PMID- 16211464 TI - Respiratory manifestations of leptospirosis: a retrospective study. AB - We retrospectively reviewed 34 consecutive patients with serologically confirmed leptospirosis admitted during years 1992-2002. Nine patients (26.5%) had respiratory symptoms on admission including cough (n = 4), shortness of breath (n = 4), cyanosis (n = 2), and hemoptysis (n = 1). Six patients had pulmonary radiographic findings including (1) diffuse, ill-defined, ground-glass density (n = 3); (2) diffuse alveolar opacities (n = 2); and (3) small nodular density (n = 1). Male/female ratio was 8/1 and mean age was 47 years. Seven patients reported their exposure source including hunting (n = 2), fishing (n = 2), fresh water swimming (n = 2), and canoeing (n = 1). All patients had fever (mean = 40.1 degrees C). Other common symptoms were headache (n = 4), vomiting (n = 3), and myalgia (n = 3). Biological abnormalities included elevated liver enzymes (n = 8), proteinuria (n = 7), lymphopenia (n = 6), hematuria (n = 5), renal failure (n = 4), anemia (n = 4), and elevated neutrophil count (n = 4). PaO(2 )was measured for 3 patients while they were breathing room air (32, 55, and 66 mmHg). Suspected diagnosis on admission included leptospirosis (n = 2), bacterial pneumonia (n = 2), intoxication, influenza, viral hepatitis, biliary tract lithiasis, and rapidly progressive glomerulonephritis (one patient each). The first serologic testing for leptospirosis was positive for 5 patients (55%). Serovar was presumptively identified for 7 patients: Australis (n = 3), Grippotyphosa (n = 2), and Icterohaemorrhagiae (n = 2). Seven patients were treated with penicillin; two patients received no antibiotics. All patients were cured. In conclusion, patients with leptospirosis may present predominantly with nonspecific pulmonary symptoms. In these patients, leptospirosis must be suspected when there is a potential exposure to rats, especially in case of high grade fever, myalgia, hepatitis, and renal abnormalities. PMID- 16211465 TI - Heterogeneous remodeling of lung vessels in idiopathic pulmonary fibrosis. AB - Recently, several reports suggest differences in the vascularization of the various histopathologic patterns of parenchymal remodeling seen in usual interstitial pneumonia (UIP). In this study, we sought to validate the importance of vascular remodeling in patients with idiopathic pulmonary fibrosis (IPF) and to examine the relationship between vascular remodeling and parenchymal remodeling or pulmonary function. Open lung biopsies were performed in 57 patients with IPF, and vascular changes in alternating areas of parenchymal remodeling (UIP histologic patterns) were studied. Quantitative analysis of the internal area, internal perimeter, wall thickness, and surrounding cellularity of medium or large pulmonary arteries, as well as their distribution according to air/parenchymal ratios, was performed. Semiquantitative analysis also was used to determine the grade of vascular occlusion. An inverse association was found between vascularization and UIP parenchymal remodeling (p < 0.05); that is, the decreased internal luminal area and perimeter as well as the increased wall thickness run in parallel with progression from alveolar collapse toward severe mural-organizing fibrosis with honeycombing. Vascular regression (diminished internal area and perimeter of vessels) was also associated with higher FEV(1), FVC, and RV values (r = 0.48, p< 0.05), reflecting a tight relationship between vascular remodeling and pulmonary function. A progressive regression of vascularization, reflected by different degrees of luminal occlusion after vascular remodeling, coincided with parenchymal remodeling (alveolar collapse, mural-organizing fibrosis, and honeycombing). This vascular regression may be responsible for the impaired wound healing and progressive fibroproliferation found in patients with IPF. Further studies are needed to determine whether this relationship is causal or consequential. PMID- 16211466 TI - The role of the corpus callosum in interhemispheric transfer of information: excitation or inhibition? AB - The corpus callosum is the major neural pathway that connects homologous cortical areas of the two cerebral hemispheres. The nature of how that interhemispheric connection is manifested is the topic of this review; specifically, does the corpus callosum serve to communicate an inhibitory or excitatory influence on the contralateral hemisphere? Several studies take the position that the corpus callosum provides the pathway through which a hemisphere or cortical area can inhibit the other hemisphere or homologous cortical area in order to facilitate optimal functional capacity. Other studies suggest that the corpus callosum integrates information across cerebral hemispheres and thus serves an excitatory function in interhemispheric communication. This review examines these two contrasting theories of interhemispheric communication. Studies of callosotomies, callosal agenesis, language disorders, theories of lateralization and hemispheric asymmetry, and comparative research are critically considered. The available research, no matter how limited, primarily supports the notion that the corpus callosum serves a predominantly excitatory function. There is evidence, however, to support both theories and the possibility remains that the corpus callosum can serve both an inhibitory and excitatory influence on the contralateral hemisphere. PMID- 16211468 TI - Ecological neuropsychology: an alternative to the deficit model for conceptualizing and serving students with learning disabilities. AB - The present paper contends that children with learning disabilities are better served when assessment and intervention are conceptualized within an ecological neuropsychology perspective than within the traditional deficit model perspective, which is the predominant approach to intervention in medical and educational settings. The deficit method conceptualizes problems as within the child, and the major consequence of this approach is that little time is spent analyzing the learning environment or other systems that might impact the child's ability to be successful in an academic setting. Therefore, rehabilitation efforts have had limited success. In contrast, ecological neuropsychology is a strength-based approach that considers the child, as well as the systems within which he/she interacts, when assessing, diagnosing, and intervening with students who are experiencing learning difficulties. PMID- 16211470 TI - Crystallizing the HIV epidemic: methamphetamine, unsafe sex, and gay diseases of the will. PMID- 16211467 TI - A meta-analysis of cognitive deficits in adults with a diagnosis of schizophrenia. AB - This review identified 1275 studies examining cognitive deficits in people with schizophrenia, published between 1990 and 2003. Data from 113 studies (4365 patients and 3429 controls) were combined in a meta-analysis carried out on the five cognitive domains of IQ, memory, language, executive function, and attention. Studies were excluded where they lacked a suitable control group or failed to present complete information. In all five cognitive domains, analysis indicated a consistent trend for patients to perform more poorly than healthy controls, with significant heterogeneity across studies. Sources of heterogeneity were analyzed and a need to ensure more appropriate composition of patient and control groups and to adopt a more refined and methodologically correct, hypothesis-driven approach was identified. PMID- 16211471 TI - Exploring the motivations and fantasies of strip club customers in relation to legal regulations. AB - Strip clubs are a popular form of adult entertainment in the contemporary United States. Strip clubs are also highly embattled entertainment venues, based on assumptions about their associations with prostitution, drug use, and "negative secondary effects" in surrounding areas, such as increased crime rates and decreased property values. Based on participant observation in five strip clubs in one city and on qualitative interviews with 30 regular male customers of those clubs, this essay seeks to challenge assumptions about the kinds of encounters sought in and purchased in such venues. Instead of visiting strip clubs out of a desire to purchase sexual release with the dancers, I found that the regular male customers were seeking an atmosphere different from both work and home, personal and sexual acceptance from women and the pleasure of a sexualized encounter without the pressures of physical performance, and a form of leisure that offered a relative degree of "safety" as well as "excitement." Further, the men's own fantasies of identity, their understandings of marriage, and their commitment to a particular kind of monogamy influenced their choice of entertainment and the pleasure that they took in their encounters with the dancers. The essay discusses these motivations and their relational aspects and assesses strip club regulation in light of these observations and findings. PMID- 16211472 TI - Timing of pubertal maturation and the onset of sexual behavior among Zimbabwe school boys. AB - The relationship between reproductive maturation and the onset of sexual behavior in boys across cultures is unclear. To explore the relative timing of pubertal events and their relationship to the onset of sexual behavior, we used data collected from 442 Zimbabwe school boys aged 12-18 years. Measures of reproductive maturation included self-reported spontaneous nocturnal emission, secondary sexual characteristics, and salivary testosterone. Behavioral measures included age at first sexual fantasies, non-coital sexual behavior, and coitus. Sigmoid interpolation curve techniques indicated a median age of first erection at 10.75 +/- 0.11 years, first sexual fantasy at 12.66 +/- 0.03 years, first spontaneous nocturnal emission at 13.02 +/- 0.03 years, and adult levels of blood testosterone at 17.2 +/- 0.7 years. First reported spontaneous nocturnal emission was significantly related to all measures of sexual behavior. Multiple regression models indicated that first reported spontaneous nocturnal emission was a stronger predictor of sexual behavior than variation in secondary sexual characteristics. In addition, testosterone was a significant predictor of sexual fantasies and intercourse, independent of first reported spontaneous nocturnal emission. These results confirm earlier findings in other populations that self reported spontaneous nocturnal emissions can be used as a marker of pubertal timing among adolescent boys. They also suggest that variation in testosterone plays a role in the onset of sexual behavior among boys, beyond its relationship to developmental timing. The behavioral mechanisms represented by the significant effects of testosterone reported here remain to be investigated. PMID- 16211473 TI - Sexual abuse in childhood and sexual dysfunction in adulthood: an Australian population-based study. AB - This study examined self-reported adult sexual functioning in individuals reporting a history of childhood sexual abuse (CSA) in a representative sample of the Australian population. A sample of 1793 persons, aged 18-59 years, were randomly selected from the electoral roll for Australian states and territories in April 2000. Respondents were interviewed about their health status and sexual experiences, including unwanted sexual experiences before the age of 16 years. More than one-third of women and approximately one-sixth of men reported a history of CSA. Women were more likely than men to report both non-penetrative and penetrative experiences of CSA. For both sexes, there was a significant association between CSA and symptoms of sexual dysfunction. In assessing the specific nature of the relationship between sexual abuse and sexual dysfunction, statistically significant associations were, in general, evident for women only. CSA was not associated with the level of physical or emotional satisfaction respondents experienced with their sexual activity. The total number of lifetime sexual partners was significantly and positively associated with CSA for females, but not for males; however, the number of sexual partners in the last year was not related to CSA. CSA in the Australian population is common and contributes to significant impairment in the sexual functioning of adults, especially women. These consequences appear not to extend to the other areas of sexual activity considered in this study. PMID- 16211474 TI - Increased high risk sexual behavior after September 11 in men who have sex with men: an Internet survey. AB - Numerous studies on the mental health effects of terrorist attacks have been published, with some reporting increases in smoking and drug and alcohol use. None have reported on changes in sexual behavior. To investigate the impact of the September 11 attacks on sexual and drug- and alcohol-using behaviors of men who have sex with men (MSM), an anonymous Internet survey was conducted to obtain information retrospectively on behavior during three month periods before and after the attacks. A total of 2,915 MSM from all 50 U.S. states completed the survey. Men who were exposed to the attacks were not differentially targeted for the survey since the online banner ad used to recruit did not mention September 11. Exposure to the attacks varied: 11.4% lost a friend or relative; 5% witnessed the attacks in person; and nearly all saw the attacks on television within one hour of their occurrence. Nearly equal proportions of men reported increases and decreases in the number of sex partners following September 11. Small, statistically significant increases in unprotected anal intercourse and alcohol use, but not illicit drug use, were found when behavior after September 11 was compared to that before the attacks. Men who lost a friend or relative in the attacks were significantly more likely to report unprotected anal intercourse, an increased number of sex partners, and increased alcohol use after September 11 than those who did not. Counseling about substance abuse and risky sexual behavior should be incorporated into trauma-related programs for adolescents and adults. PMID- 16211475 TI - Sex differences in affective responses to homoerotic stimuli: evidence for an unconscious bias among heterosexual men, but not heterosexual women. AB - Antigay bias is a well-documented social problem among heterosexual men, though heterosexual women display a lesser tendency toward this bias. Startle eye blink has been established as a valid measure of the affective component of antigay bias in heterosexual men. In the current study, a sample of 91 heterosexual women and 87 heterosexual men were exposed to a variety of sexual photographic stimuli accompanied by startle probes. Heterosexual men who expressed more bias against gay men using a social distance measure (i.e., discomfort with being in close quarters with a gay man) displayed a startle response consistent with greater negative affect (e.g., fear and disgust) toward gay male stimuli, while those with less self-reported antigay bias did not display a physiological bias against gay men, and none of these men showed a relationship between bias against lesbians and physiological responses while viewing lesbian images. There were no such physiological manifestations of antigay bias in heterosexual women while viewing lesbian or gay male images, even among those who self-reported such bias. It appears that heterosexual women do not tend to have the same affective response toward homosexuals that some heterosexual men experience. PMID- 16211476 TI - Appetitive responses to sexual stimuli are attenuated in individuals with low levels of sexual desire. AB - Despite the high prevalence of sexual desire disorders, little is known about their biological underpinnings in humans. Animal studies suggest that dopamine is involved in appetitive sexual behavior; thus, one aim of this study was to elucidate that relationship in humans. This study used measurement of the acoustic startle response (ASR) and prepulse inhibition of the startle response (PPI) as psychophysiological indicators of changes in motivational states to assess the potential relation between sexual desire and appetitive motivation in humans. Responses to sexually provocative stimuli consisting of single nude men and single nude women in a sample of 153 participants (77 men, 76 women) were assessed. The results indicated that ASR was attenuated after exposure to appetitive stimuli (i.e., sexually provocative pictures of attractive individuals) to a greater extent among participants with higher levels of sexual desire, as measured by the Sexual Desire Inventory-2 (Spector, I. P., Carey, M. P., & Steinberg, L. (1996). Journal of Sex & Marital Therapy, 22, 175-190). In addition, PPI was inversely associated with subjective ratings across stimuli such that greater subjective levels of desire were correlated with lower levels of PPI. In general, these results suggest that individuals with lower levels of sexual desire may have a diminished physiological response to appetitive sexual stimuli. PMID- 16211477 TI - Stability and change in sexual practices among first-year Australian university students (1990-1999). AB - A questionnaire on sexual attitudes and behavior was administered to first-year students at Macquarie University in Sydney every year from 1990 to 1999 (N = 4295 aged 18-19; 72.5% female). Responses to questions about experience of different sexual practices (tongue kissing, oral sex, and vaginal intercourse) with regular and casual partners were analyzed for trends. Over half of the students each year (on average 64% of the men, 57% of the women) had experience of oral sex or vaginal intercourse. More male than female students reported experience of each practice, especially with casual partners. Rates for female students increased significantly over the 10-year period for all practices except tongue kissing with a regular partner and vaginal sex with a casual partner; rates for male students were apparently steady. Results are consistent with evidence from other sources of an increase in the acceptability of oral sex (both fellatio and cunnilingus) in recent decades and of increasing similarity between young men's and women's reports of sexual experience. PMID- 16211478 TI - Sexual orientation in United States and Canadian college students. AB - Based on a sample of nearly 8,000 college students, this study was undertaken to determine the prevalence of non-heterosexuality using four different measures of sexual orientation: a self-identity measure, an attraction measure, a fantasy measure, and an experiential measure. Over 97% of both males and females labeled themselves as heterosexual, with the proportion of homosexuals and bisexuals combined constituting nearly 3% of the male sample and about 2% of the female sample. Roughly 80-85% of both sexes would be classified as exclusively heterosexual, with about 10% of both sexes having at least half of their sexual fantasies involving same-sex partners. In terms of sexual experiences, about 5% of non-virgin males and 0.5% of non-virgin females reported that all of their experiences had involved same-sex partners. After examining each of the sexual orientation measures separately, we compared them to one another. These comparisons revealed an unsettling number of apparent contradictions in the responses given by individual participants (e.g., participants who stated that they were homosexual but that all of their fantasies involved members of the opposite sex). By eliminating participants who provided these apparent contradictory responses, a post-hoc refined self-identified measure of sexual orientation was created. This refinement had essentially no effect on our estimates of the proportion of college students who were identified as heterosexual or homosexual, thus suggesting that the inconsistencies were largely due to random response error. Boosting our confidence in the post-hoc refined measure, we found that it exhibited slightly stronger links with several established childhood correlates of sexual orientation (such as the enjoyment of collecting dolls and playing dress-up) than was true for the original self identified sexual orientation. PMID- 16211480 TI - Resolution-enhanced base-type-edited HCN experiment for RNA. AB - New base-type-edited transverse-relaxation optimized CT-HCN(C) experiments are presented that yield intra-base and sugar-to-base correlations for 13C-15N labeled RNA. High spectral resolution in the 13C and 15N dimensions is achieved by constant time (CT) frequency editing. A spectral editing filter applied during the CT 15N labeling period separates the correlation peaks arising from G/U and A/C nucleotide bases. This provides the increased spectral resolution required to unambiguously connect the 1H and 13C resonances in sugar and base moieties of RNA nucleotides. In addition, the experiment allows base type identification for each residue, and therefore presents an attractive spectroscopic alternative to nucleotide-specific isotope labeling. Application to a 33-nucleotide RNA aptamer demonstrates the performance of the new pulse scheme. PMID- 16211481 TI - Interpreting dynamically-averaged scalar couplings in proteins. AB - The experimental determination of scalar three-bond coupling constants represents a powerful method to probe both the structure and dynamics of proteins. The detailed structural interpretation of such coupling constants is usually based on Karplus relationships, which allow the measured couplings to be related to the torsion angles of the molecules. As the measured couplings are sensitive to thermal fluctuations, the parameters in the Karplus relationships are better derived from ensembles representing the distributions of dihedral angles present in solution, rather than from single conformations. We present a method to derive such parameters that uses ensembles of conformations determined through dynamic ensemble refinement--a method that provides structural ensembles that simultaneously represent both the structure and the associated dynamics of a protein. PMID- 16211483 TI - Residue specific ribose and nucleobase dynamics of the cUUCGg RNA tetraloop motif by MNMR 13C relaxation. AB - The dynamics of the nucleobase and the ribose moieties in a 14-nt RNA cUUCGg hairpin-loop uniformly labeled with 13C and 15N were studied by 13C spin relaxation experiments. R1, R1rho and the 13C-[1H] steady-state NOE of C6 and C1' in pyrimidine and C8 and C1' in purine residues were obtained at 298 K. The relaxation data were analyzed by the model-free formalism to yield dynamic information on timescales of pico-, nano- and milli-seconds. An axially symmetric diffusion tensor with an overall rotational correlation time tau(c) of 2.31 +/- 0.13 ns and an axial ratio of 1.35 +/- 0.02 were determined. Both findings are in agreement with hydrodynamic calculations. For the nucleobase carbons, the validity of different reported 13C chemical shift anisotropy values (Stueber, D. and Grant, D. M., 2002 J. Am. Chem. Soc. 124, 10539-10551; Fiala et al., 2000 J. Biomol. NMR 16, 291-302; Sitkoff, D. and Case, D. A., 1998 Prog. NMR Spectroscopy 32, 165-190) is discussed. The resulting dynamics are in agreement with the structural features of the cUUCGg motif in that all residues are mostly rigid (0.82 < S2 < 0.96) in both the nucleobase and the ribose moiety except for the nucleobase of U7, which is protruding into solution (S2 = 0.76). In general, ribose mobility follows nucleobase dynamics, but is less pronounced. Nucleobase dynamics resulting from the analysis of 13C relaxation rates were found to be in agreement with 15N relaxation data derived dynamic information (Akke et al., 1997 RNA 3, 702-709). PMID- 16211482 TI - A new amide proton R1rho experiment permits accurate characterization of microsecond time-scale conformational exchange. AB - A new off-resonance spin-lock experiment to record relaxation dispersion profiles of amide protons is presented. The sensitivity-enhanced HSQC-type sequence is designed to minimize the interference from cross-relaxation effects and ensure that the dispersion profiles in the absence of micros-ms time-scale dynamics are flat. Toward this end (i) the proton background is eliminated by sample deuteration (Ishima et al., 1998), (ii) 1H spin lock is applied to two-spin modes 2(H(x)Sin theta + H(z)Cos theta) N(z), and (iii) the tilt angle theta approximately 35 degrees is maintained throughout the series of measurements (Desvaux et al. Mol. Phys., 86 (1995) 1059). The relaxation dispersion profiles recorded in this manner sample a wide range of effective rf field strengths (up to and in excess of 20 kHz) which makes them particularly suitable for studies of motions on the time scale < or = 100 micros. The new experiment has been tested on the Ca2+-loaded regulatory domain of cardiac troponin C. Many residues show pronounced dispersions with remarkably similar correlation times of approximately 30 micros. Furthermore, these residues are localized in the regions that have been previously implicated in conformational changes (Spyracopoulos et al. Biochemistry, 36 (1997) 12138). PMID- 16211484 TI - Triple-resonance methods for complete resonance assignment of aromatic protons and directly bound heteronuclei in histidine and tryptophan residues. AB - A set of three experiments is described which correlate aromatic resonances of histidine and tryptophan residues with amide resonances in 13C/15N-labelled proteins. Provided that backbone 1H and 15N positions of the sequentially following residues are known, this results in sequence-specific assignment of histidine 1H(delta2)/13C(delta2) and 1H(epsilon1)/13C(epsilon1) as well as tryptophan 1H(delta1)/13C(delta1), 1H(zeta2)/13C(zeta2), 1H(eta2)/13C(eta2), 1H(epsilon3)/13C(epsilon3), 1H(zeta3)/13C(zeta3) and 1H(epsilon1)/15N(epsilon1) chemical shifts. In the reverse situation, these residues can be located in the 1H-(15)N correlation map to facilitate backbone assignments. It may be chosen between selective versions for either of the two amino acid types or simultaneous detection of both with complete discrimination against phenylalanine or tyrosine residues in each case. The linkages between delta-proton/carbon and the remaining aromatic as well as backbone resonances do not rely on through-space interactions, which may be ambiguous, but exclusively employ one-bond scalar couplings for magnetization transfer instead. Knowledge of these aromatic chemical shifts is the prerequisite for the analysis of NOESY spectra, the study of protein-ligand interactions involving histidine and tryptophan residues and the monitoring of imidazole protonation states during pH titrations. The new methods are demonstrated with five different proteins with molecular weights ranging from 11 to 28 kDa. PMID- 16211487 TI - 1H, 15N and 13C resonance assignments of the putative Bet v 1 family protein At1g24000.1 from Arabidopsis thaliana. PMID- 16211486 TI - NMR Assignment Reveals an alpha-Helical Fold for the F-Actin Binding Domain of Human Bcr-Abl/c-Abl. PMID- 16211485 TI - Solution structure of the mouse enhancer of rudimentary protein reveals a novel fold. PMID- 16211488 TI - Complete assignment and secondary structure of the Brazil nut allergen Ber e 1. PMID- 16211489 TI - Sequential resonance assignment of the human BMP type II receptor extracellular domain. PMID- 16211490 TI - Backbone NMR assignment of the C-terminal haemopexin-like domain (HPLD) of human matrix metalloproteinase MMP-13. PMID- 16211491 TI - 1H, 15N and 13C resonance assignments of a protein involved in the autophagy process, At4g21980.1 from Arabidopsis thaliana. PMID- 16211492 TI - 1H, 15N and 13C resonance assignments of the heme-binding protein murine p22HBP. PMID- 16211493 TI - Backbone NMR assignment of the human E2 ubiquitin conjugating enzyme UbcH5alpha (F72K,F82S) double mutant. PMID- 16211494 TI - 1H, 15N, and 13C chemical shift assignments of the human Sulfiredoxin (hSrx). PMID- 16211495 TI - NMR assignment of the apo and peptide-bound SH2 domain from the Rous sarcoma viral protein Src. PMID- 16211496 TI - NMR assignment of human ubiquitin conjugating enzyme Ubc7. PMID- 16211497 TI - 1H, 13C, and 15N sequence-specific resonance assignment and secondary structure of Plasmodium falciparum thioredoxin. PMID- 16211501 TI - Structural genomics of minimal organisms and protein fold space. AB - The initial aim of the Berkeley Structural Genomics Center is to obtain a near complete structural complement of two minimal organisms, closely related pathogens Mycoplasma genitalium and M. pneumoniae. The former has fewer than 500 genes and the latter fewer than 700 genes. To achieve this goal, the current protein targets have been selected starting with those predicted to be most tractable and likely to yield new structural and functional information. During the past 3 years, the semi-automated structural genomics pipeline has been set up from cloning, expression, purification, and ultimately to structural determination. The results from the pipeline substantially increased the coverage of the protein fold space of M. pneumoniae and M. genitalium. Furthermore, about 1/2 of the structures of 'unique' protein sequences revealed new and novel folds, and over 2/3 of the structures of previously annotated 'hypothetical proteins' inferred their molecular functions. PMID- 16211502 TI - Protein production and crystallization at the joint center for structural genomics. AB - By definition, structural genomics centers must be able to address a large number of diverse protein targets. The methods developed should permit parallel and cost effective processing while allowing for the diverse nature of proteins. Our approach to this problem is a multi-tiered effort where targets are characterized and categorized by behavior and processed in parallel by appropriate methods. The Joint Center for Structural Genomics (JCSG) has applied this tactic to create a fully integrated and scaleable structure determination pipeline. Highlights of the development of the current pipeline for protein production and crystallization are presented here. PMID- 16211503 TI - Structural genomics of eukaryotic targets at a laboratory scale. AB - Structural genomics programs are distributed worldwide and funded by large institutions such as the NIH in United-States, the RIKEN in Japan or the European Commission through the SPINE network in Europe. Such initiatives, essentially managed by large consortia, led to technology and method developments at the different steps required to produce biological samples compatible with structural studies. Besides specific applications, method developments resulted mainly upon miniaturization and parallelization. The challenge that academic laboratories faces to pursue structural genomics programs is to produce, at a higher rate, protein samples. The Structural Biology and Genomics Department (IGBMC - Illkirch - France) is implicated in a structural genomics program of high eukaryotes whose goal is solving crystal structures of proteins and their complexes (including large complexes) related to human health and biotechnology. To achieve such a challenging goal, the Department has established a medium-throughput pipeline for producing protein samples suitable for structural biology studies. Here, we describe the setting up of our initiative from cloning to crystallization and we demonstrate that structural genomics may be manageable by academic laboratories by strategic investments in robotic and by adapting classical bench protocols and new developments, in particular in the field of protein expression, to parallelization. PMID- 16211504 TI - Methods and results for semi-automated cloning using integrated robotics. AB - The Joint Center for Structural Genomics (JCSG) has emphasized automation and parallel processing approaches. Here, we describe automated methods used across the cloning process with results from JCSG projects. The protocols for PCR, restriction digests and ligations, as well as for gel electrophoresis and microtiter plate assays have all been automated. The system has the capacity to routinely process 384 clones a week. This throughput can adequately supply our expression and purification pipeline with expression-ready clones, including novel targets and truncations. The utility of our system is demonstrated by our results from three diverse projects. In summary, 94% of the PCR amplicons generated to date have been successfully cloned and verified by sequencing (83% of the total attempted targets). Our results demonstrate the capabilities of this robotic platform to provide an avenue to high-throughput cloning which requires little manpower and is rapid and cost-effective while providing insights for method optimization. PMID- 16211505 TI - Incorporation of selenomethionine into induced intracytoplasmic membrane proteins of Rhodobacter species. AB - Efficient multiple- or single-wavelength anomalous dispersion (MAD/SAD) techniques that use tunable X-ray sources at third-generation synchrotrons exploit the anomalous scattering of certain heavy atoms for determination of experimental phases. Development of methods for the in vivo substitution of methionine by selenomethionine (SeMet) has revolutionized the process for determination of structures of soluble proteins in recent years. Herein, we report methods for biosynthetic incorporation of SeMet into induced intracytoplasmic membrane proteins of two species of the Rhodobacter genus of purple non-sulfur photosynthetic bacteria. Amino acid analysis of a membrane protein complex that was purified to homogeneity determined that the extent of SeMet incorporation was extensive and approached quantitative replacement. Diffraction-quality crystals were obtained from SeMet-labeled membrane proteins purified from 2 l of culture. These methods augment the potential utility of photosynthetic bacteria and their inducible membrane systems for the production of foreign membrane proteins for structure determination. PMID- 16211506 TI - Enhanced expression and purification of membrane proteins by SUMO fusion in Escherichia coli. AB - Severe acute respiratory syndrome coronavirus (SARS-CoV) membrane protein and 5 lipoxygenase-activating protein (FLAP) are among a large number of membrane proteins that are poorly expressed when traditional expression systems and methods are employed. Therefore to efficiently express difficult membrane proteins, molecular biologists will have to develop novel or innovative expression systems. To this end, we have expressed the SARS-CoV M and FLAP proteins in Escherichia coli by utilizing a novel gene fusion expression system that takes advantage of the natural chaperoning properties of the SUMO (small ubiquitin-related modifier) tag. These chaperoning properties facilitate proper protein folding, which enhances the solubility and biological activity of the purified protein. In addition to these advantages, we found that SUMO Protease 1, can cleave the SUMO fusion high specificity to generate native protein. Herein, we demonstrate that the expression of FLAP and SARS-CoV membrane proteins are greatly enhanced by SUMO fusions in E. coli. PMID- 16211507 TI - Recent advances in GFP folding reporter and split-GFP solubility reporter technologies. Application to improving the folding and solubility of recalcitrant proteins from Mycobacterium tuberculosis. AB - We have improved our green fluorescent protein (GFP) folding reporter technology [Waldo et al., (1999) Nat. Biotechnol. 17, 691-695] to evolve recalcitrant proteins from Mycobacterium tuberculosis. The target protein is inserted into the scaffolding of the GFP, eliminating false-positive artifacts caused by expression of truncated protein variants from internal cryptic ribosome binding sites in the target RNA. In parallel, we have developed a new quantitative fluorescent protein tagging and detection system based on micro-domains of GFP. This split-GFP system, which works both in vivo and in vitro, is amenable to high-throughput assays of protein expression and solubility [Cabantous et al., (2005) Nat. Biotechnol. 23, 102-107]. Together, the GFP folding reporter and split-GFP technologies offer a comprehensive system for manipulating and improving protein folding and solubility. PMID- 16211509 TI - High-throughput limited proteolysis/mass spectrometry for protein domain elucidation. AB - High-resolution structural information is important for improving our understanding of protein function in vitro and in vivo and providing information to enable drug discovery. The process leading to X-ray structure determination is often time consuming and labor intensive. It requires informed decisions in expression construct design, expression host selection, and strategies for protein purification, crystallization and structure determination. Previously published studies have demonstrated that compact globular domains defined by limited proteolysis represent good candidates for production of diffraction quality crystals [1-7]. Integration of mass spectrometry and proteolysis experiments can provide accurate definition of domain boundaries at unprecedented rates. We have conducted a critical evaluation of this approach with 400 target proteins produced by SGX (Structural GenomiX, Inc.) for the New York Structural GenomiX Research Consortium (NYSGXRC; http://www.nysgxrc.org) under the auspices of the National Institute of General Medical Sciences Protein Structure Initiative (http://www.nigms.nih.gov/psi). The objectives of this study were to develop parallel/automated protocols for proteolytic digestion and data acquisition for multiple proteins, and to carry out a systematic study to correlate domain definition via proteolysis with outcomes of crystallization and structure determination attempts. Initial results from this work demonstrate that proteins yielding diffraction quality crystals are typically resistant to proteolysis. Large-scale sub cloning and subsequent testing of expression, solubility, and crystallizability of proteolytically defined truncations is currently underway. PMID- 16211508 TI - Salvaging Pyrococcus furiosus protein targets at SECSG. AB - Proteins derived from the coding regions of Pyrococcus furiosus are targets for three-dimensional X-ray and NMR structure determination by the Southeast Collaboratory for Structural Genomics (SECSG). Of the 2,200 open reading frames (ORFs) in this organism, 220 protein targets were cloned and expressed in a high throughput (HT) recombinant system for crystallographic studies. However, only 96 of the expressed proteins could be crystallized and, of these, only 15 have led to structures. To address this issue, SECSG has recently developed a two-tier approach to protein production and crystallization. In this approach, tier-1 efforts are focused on producing protein for new Pfu(italics?) targets using a high-throughput approach. Tier-2 protein production efforts support tier-1 activities by (1) producing additional protein for further crystallization trials, (2) producing modified protein (further purification, methylation, tag removal, selenium labeling, etc) as required and (3) serving as a salvaging pathway for failed tier-1 proteins. In a recent study using this two-tiered approach, nine structures were determined from a set of 50 Pfu proteins, which failed to produce crystals suitable for X-ray diffraction analysis. These results validate this approach and suggest that it has application to other HT crystal structure determination applications. PMID- 16211510 TI - High-throughput protein production for X-ray crystallography and use of size exclusion chromatography to validate or refute computational biological unit predictions. AB - The production of large numbers of highly purified proteins for X-ray crystallography is a significant bottleneck in structural genomics. At the Joint Center for Structural Genomics (JCSG; http://www.jcsg.org), specific automated protein expression, purification, and analytical methods are being utilized to study the proteome of Thermotoga maritima. Anion exchange and size exclusion chromatography (SEC), intended for the production of highly purified proteins, have been automated and the procedures are described here in detail. Analytical SEC has been included as a standard quality control test. A biological unit (BU) is the macromolecule that has been proven or is presumed to be functional. Correct assignment of BUs from protein structures can be difficult. BU predictions obtained via the Protein Quaternary Structure file server (PQS; http://pqs.ebi.ac.uk/) were compared to SEC data for 16 representative T. maritima proteins whose structures were solved at the JCSG, revealing an inconsistency in five cases. Herein, we report that SEC can be used to validate or disprove PQS-derived oligomeric models. A substantial amount of associated SEC and structural data should enable us to use certain PQS parameters to gauge the accuracy of these computational models and to generally improve their predictions. PMID- 16211511 TI - High-throughput purification and quality assurance of Arabidopsis thaliana proteins for eukaryotic structural genomics. AB - The Center for Eukaryotic Structural Genomics (CESG) has established procedures for the purification of Arabidopsis proteins in a high-throughput mode. Recombinant proteins were fused with (His)(6)-MBP tags at their N-terminus and expressed in Escherichia coli. Using an automated AKTApurifier system, fusion proteins were initially purified by immobilized metal affinity chromatography (IMAC). After cleavage of (His)(6)-MBP tags by TEV protease, (His)(6)-MBP tags were separated from target proteins by a subtractive 2nd IMAC. As a part of quality assurance, all purified proteins were subjected to MALDI-TOF and ESI mass spectrometry to confirm target identity and integrity, and determine incorporation of seleno-methionine (SeMet) and (15)N and (13)C isotopes. The protocols have been used successfully to provide high quality proteins that are suitable for structural studies by X-ray crystallography and NMR. PMID- 16211512 TI - Comparison of small- and large-scale expression of selected Pyrococcus furiosus genes as an aid to high-throughput protein production. AB - As the natural extension of the genomic sequencing projects, the goal of the various world-wide Structural Genomics projects is development of techniques for high throughput (HTP) cloning, protein overexpression, purification and structural determination, with the ultimate goal of determining all possible protein structures. Rapid (small-scale) screening of potential expression clones under different growth conditions is presumed to be possible and a viable way to increase throughput of protein expression. In order to test the utility of screening for soluble, heterologous protein expression, we have compared the production of recombinant proteins on a small scale (1 ml cultures in 96-well plates) in Escherichia coli under two growth conditions [a rich medium and a defined (minimal) medium] using an enzyme-linked immunosorbent assay (ELISA) against the affinity tag, with the amount of recombinant protein produced during the large-scale (500 ml) growth of E. coli. The large-scale expression products were examined after a single step affinity purification by visualization on SDS PAGE gels. Of the open reading frames that were successfully expressed on the 1 ml scale as judged by immunodetection, 80% of them successfully scaled-up to 500 ml in a rich medium and 81% of them scaled-up in a defined medium. This is significantly higher than would be expected by a randomly selected expression condition and validates the use of small-scale expression as a screening tool for more efficient protein production. PMID- 16211514 TI - Large-scale transient transfection of mammalian cells: a newly emerging attractive option for recombinant protein production. AB - Mammalian expression systems have an undisputed long-standing and very successful history for the generation of recombinant proteins, mainly as biopharmaceuticals. However, for use as 'tool proteins' in, e.g. assay development and screening, for structure elucidation and as antigens these expression systems were generally regarded as being cumbersome, tedious and expensive. This bias has largely been overcome with the very recent development of large-scale transient transfection (LST) approaches. Especially the HEK.EBNA expression system described here has contributed significantly to this success. The simplicity and speed of this approach compares well with expression trials using the widely applied Baculovirus/insect cell system. In addition, proteins generated in mammalian cells are usually correctly folded, fully processed and functionally active. PMID- 16211513 TI - Large-scale expression and purification of a G-protein-coupled receptor for structure determination -- an overview. AB - Structure determination of G-protein-coupled receptors and other applications, such as nuclear magnetic resonance studies, require milligram quantities of purified, functional receptor protein on a regular basis. We present an overview on expression and purification studies with a receptor for neurotensin. Functional expression in Escherichia coli and an automated two-column purification routine allow ongoing crystallization experiments and studies on receptor-bound ligands. PMID- 16211515 TI - Rapid isolation of single-chain antibodies for structural genomics. AB - High throughput approaches to structural genomics requires expression, purification, and crystallization of proteins derived from predicted open reading frames cloned into a host organism, typically E. coli. Early results from this approach suggest that the success rate of obtaining well diffracting crystals from eukaryotic proteins is disappointingly low. A proven method of improving the odds of crystallization is formation of a complex with a conformation-stabilizing partner of known structure that is easily crystallized. Such complexes are also able to engage in different crystal contacts than the original protein by itself. Fab fragments derived from monoclonal antibodies have been successfully used for this purpose for a variety of proteins, however conventional methods for the isolation of monoclonal antibodies from hybridomas are time consuming and expensive. We are exploring the use of phage display to generate recombinant antibodies to target proteins that can be used to obtain co-complexes to facilitate crystallization and structural determination. We are using a large, human single-chain Fv (scFv) library to select for antibodies that bind to a panel of Leishmania major target proteins. Thirteen out of 16 target proteins yielded good binders after three rounds of enrichment. A total of 55 distinct scFvs were identified, with five targets each yielding at least five different scFvs. Individual clones were analyzed for binding specificity and soluble scFv can be readily produced and purified via the appended His(6) epitope tag. Using immunoaffinity chromatography, eight scFv target protein pairs were identified that exhibit stable complex formation and are suitable for co-crystallization trials. PMID- 16211516 TI - On-column protein refolding for crystallization. AB - One major bottleneck in protein production in Escherichia coli for structural genomics projects is the formation of insoluble protein aggregates (inclusion bodies). The efficient refolding of proteins from inclusion bodies is becoming an important tool that can provide soluble native proteins for structural and functional studies. Here we report an on-column refolding method established at the Berkeley Structural Genomics Center (BSGC). Our method is a combination of an 'artificial chaperone-assisted refolding' method previously proposed and affinity chromatography to take advantage of a chromatographic step: less time-consuming, no filtration or concentration, with the additional benefit of protein purification. It can be easily automated and formatted for high-throughput process. PMID- 16211517 TI - Designing a high throughput refolding array using a combination of the GroEL chaperonin and osmolytes. AB - Although GroE chaperonins and osmolytes had been used separately as protein folding aids, combining these two methods provides a considerable advantage for folding proteins that cannot fold with either osmolytes or chaperonins alone. This technique rapidly identifies superior folding solution conditions for a broad array of proteins that are difficult or impossible to fold by other methods. While testing the broad applicability of this technique, we have discovered that osmolytes greatly simplify the chaperonin reaction by eliminating the requirement for the co-chaperonin GroES which is normally involved in encapsulating folding proteins within the GroEL-GroES cavity. Therefore, combinations of soluble or immobilized GroEL, osmolytes and ATP or even ADP are sufficient to refold the test proteins. The first step in the chaperonin/osmolyte process is to form a stable long-lived chaperonin-substrate protein complex in the absence of nucleotide. In the second step, different osmolyte solutions are added along with nucleotides, thus forming a 'folding array' to identify superior folding conditions. The stable chaperonin-substrate protein complex can be concentrated or immobilized prior to osmolyte addition. This procedure prevents off pathway aggregation during folding/refolding reactions and more importantly allows one to refold proteins at concentrations (approximately mg/ml) that are substantially higher than the critical aggregation concentration for given protein. This technique can be used for successful refolding of proteins from purified inclusion bodies. Recently, other investigators have used our chaperonin/osmolyte method to demonstrate that a mutant protein that misfolds in human disease can be rescued by GroEL/osmolyte system. Soluble or immobilized GroEL can be easily removed from the released folded protein using simple separation techniques. The method allows for isolation of folded monomeric or oligomeric proteins in quantities sufficient for X-ray crystallography or NMR structural determinations. PMID- 16211518 TI - InsectDirect System: rapid, high-level protein expression and purification from insect cells. AB - A fundamental challenge in high-throughput (HT) expression screening is to rapidly identify the appropriate expression system for many targets in parallel. Known or unknown open reading frames (ORFs) are typically amplified by PCR and then cloned into a variety of vectors, producing recombinants used to direct target protein expression in Escherichia coli, insect cells, mammalian cells, or yeast. To facilitate rapid expression and purification in Spodoptera insect cells (Sf9), we developed transient expression vectors that include an enterokinase cleavage site immediately upstream of a ligation-independent cloning site (Ek/LIC). We also developed a high-efficiency insect cell transfection reagent, and automation-compatible fusion protein purification system for insect cells to facilitate expression screening and protein production. Positive clones identified from the small-scale screening were subjected to a larger scale production. Using this InsectDirect approach, we successfully expressed milligram quantities of different human proteins including heat shock proteins, phospholipases, and protein kinases. PMID- 16211519 TI - Automatic classification and pattern discovery in high-throughput protein crystallization trials. AB - Conceptually, protein crystallization can be divided into two phases search and optimization. Robotic protein crystallization screening can speed up the search phase, and has a potential to increase process quality. Automated image classification helps to increase throughput and consistently generate objective results. Although the classification accuracy can always be improved, our image analysis system can classify images from 1,536-well plates with high classification accuracy (85%) and ROC score (0.87), as evaluated on 127 human classified protein screens containing 5,600 crystal images and 189,472 non crystal images. Data mining can integrate results from high-throughput screens with information about crystallizing conditions, intrinsic protein properties, and results from crystallization optimization. We apply association mining, a data mining approach that identifies frequently occurring patterns among variables and their values. This approach segregates proteins into groups based on how they react in a broad range of conditions, and clusters cocktails to reflect their potential to achieve crystallization. These results may lead to crystallization screen optimization, and reveal associations between protein properties and crystallization conditions. We also postulate that past experience may lead us to the identification of initial conditions favorable to crystallization for novel proteins. PMID- 16211520 TI - Spectroscopic imaging of protein crystals in crystallization drops. AB - Automatic imaging and scoring of crystallization drops is an essential step in high-throughput crystallography. Presently, white-light images of crystallization drops are acquired robotically and the images are analyzed and scored using pattern recognition algorithms. However, the scoring part remains unreliable as crystals and microcrystals are not always recognized by existing feature extraction and recognition algorithms. We propose a fundamental shift in crystal monitoring through spectroscopic imaging of crystallization drops. This method converts the problem of automatic crystal detection from one of pattern recognition into one of intensity (concentration) analysis. The latter can be more robust and reliable. PMID- 16211521 TI - Shotgun crystallization strategy for structural genomics II: crystallization conditions that produce high resolution structures for T. maritima proteins. AB - Currently, 119 high resolution structures of Thermotoga maritima proteins have been determined by the Joint Center for Structural Genomics (JCSG, www.jcsg.org). Sixty-seven of these were solved using the first implementation of the multi tiered crystallization strategy at the JCSG for the efficient crystallization of large numbers of protein targets. Previously, we reported the analysis of all proteins crystallized using this multi-tiered strategy [Lesley, S.A. et al. (2002) Proc. Natl. Acad. Sci. USA 99, 11664-11669; Page, R. et al. (2003) Acta Crystallogr. D Biol. Crystallogr. 59, 1028-1037]. Here, we extend the analysis and describe the crystallization characteristics of those proteins that produced diffraction quality crystals, ultimately resulting in high resolution structures. First, we found that over 77% (52) of the crystals used for structure determination were produced directly from high-throughput coarse screens, indicating that less than one quarter of the crystals (15) required fine screening. In addition, as observed for the proteome screen [Page, R. et al. (2003) Acta Crystallogr. D Biol. Crystallogr. 59, 1028-1037], the majority of conditions that produced crystals for natively expressed proteins, whose structures have been determined, were distinct from those of their more extensively purified and selenomethionine-labeled counterparts. Finally, 99% of the proteins whose structures were solved crystallized in conditions contained in the JCSG Minimal Core Screen [Page, R. et al. (2003) Acta Crystallogr. D Biol. Crystallogr. 59, 1028-1037; Page, R. and Stevens, R.C. (2004) Methods 34, 373 389], a set of 67 conditions previously identified as those most likely to produce crystals of a diverse set of proteins, confirming its success for rapid identification of proteins with a natural propensity to crystallize. PMID- 16211523 TI - New York-Structural GenomiX Research Consortium (NYSGXRC): a large scale center for the protein structure initiative. AB - Structural GenomiX, Inc. (SGX), four New York area institutions, and two University of California schools have formed the New York Structural GenomiX Research Consortium (NYSGXRC), an industrial/academic Research Consortium that exploits individual core competencies to support all aspects of the NIH-NIGMS funded Protein Structure Initiative (PSI), including protein family classification and target selection, generation of protein for biophysical analyses, sample preparation for structural studies, structure determination and analyses, and dissemination of results. At the end of the PSI Pilot Study Phase (PSI-1), the NYSGXRC will be capable of producing 100-200 experimentally determined protein structures annually. All Consortium activities can be scaled to increase production capacity significantly during the Production Phase of the PSI (PSI-2). The Consortium utilizes both centralized and de-centralized production teams with clearly defined deliverables and hand-off procedures that are supported by a web-based target/sample tracking system (SGX Laboratory Information Data Management System, LIMS, and NYSGXRC Internal Consortium Experimental Database, ICE-DB). Consortium management is provided by an Executive Committee, which is composed of the PI and all Co-PIs. Progress to date is tracked on a publicly available Consortium web site (http://www.nysgxrc.org) and all DNA/protein reagents and experimental protocols are distributed freely from the New York City Area institutions. In addition to meeting the requirements of the Pilot Study Phase and preparing for the Production Phase of the PSI, the NYSGXRC aims to develop modular technologies that are transferable to structural biology laboratories in both academe and industry. The NYSGXRC PI and Co-PIs intend the PSI to have a transforming effect on the disciplines of X-ray crystallography and NMR spectroscopy of biological macromolecules. Working with other PSI-funded Centers, the NYSGXRC seeks to create the structural biology laboratory of the future. Herein, we present an overview of the organization of the NYSGXRC and describe progress toward development of a high-throughput Gene- >Structure platform. An analysis of current and projected consortium metrics reflects progress to date and delineates opportunities for further technology development. PMID- 16211522 TI - Problems in obtaining diffraction-quality crystals of hetero-oligomeric integral membrane proteins. AB - The major barrier responsible for the slow pace of structure determination of integral membrane proteins is the difficulty of crystallizing detergent solubilized hydrophobic proteins, particularly hetero-oligomeric integral membrane proteins. For the latter class of multi-subunit proteins, we have encountered the following problems in addition to the ubiquitous problem of detergent compatibility: (i) instability caused by over-purification that results in delipidation; (ii) protease activity degrading exposed loops and termini of subunits of the complex that could not be inhibited; (iii) poor protein-protein contacts presumably arising from masking by the detergent micelle. Problem (i) could be ameliorated in crystallization of the cytochrome b(6)f complex by augmenting the delipidated complex with synthetic lipid. Problem (ii) has not been solved. Problem (iii) has been solved in other systems by the use of monoclonal antibodies (or other protein ligands) to increase the probability of protein-protein contacts. In the case of the complex formed by the cobalamin and colicin receptor, BtuB, and the receptor binding domain of colicin E3, the latter served as a ligand for protein-protein contacts that facilitated crystallization. PMID- 16211524 TI - Protein production and crystallization at SECSG -- an overview. AB - Using a high degree of automation, the Southeast Collaboratory for Structural Genomics (SECSG) has developed high throughput pipelines for protein production, and crystallization using a two-tiered approach. Primary, or tier-1, protein production focuses on producing proteins for members of large Pfam families that lack a representative structure in the Protein Data Bank. Target genomes are Pyrococcus furiosus and Caenorhabditis elegans. Selected human proteins are also under study. Tier-2 protein production, or target rescue, focuses on those tier-1 proteins, which either fail to crystallize or give poorly diffracting crystals. This two tier approach is more efficient since it allows the primary protein production groups to focus on the production of new targets while the tier-2 efforts focus on providing additional sample for further studies and modified protein for structure determination. Both efforts feed the SECSG high throughput crystallization pipeline, which is capable of screening over 40 proteins per week. Details of the various pipelines in use by the SECSG for protein production and crystallization, as well as some examples of target rescue are described. PMID- 16211525 TI - Fluid balance in CRRT: a call to attention! PMID- 16211526 TI - Management of fluid balance in CRRT: a technical approach. AB - BACKGROUND: The possibility of making fluid balance errors during continuous renal replacement therapy has been identified since the beginning of this modality of treatment. The advent of automated machines has partially overcome this problem. Nevertheless, there are conditions and operation modes in which the potential for fluid balance errors is still present. OBJECTIVE: To analyse fluid balance management in CRRT therapies across a range of currently marketed machine. METHODS: The tests were conducted in vitro, utilizing saline solution for the blood circuit and regular dialysate/reinfusate for the dialysate/reinfusion circuit. The methodology used was based on the voluntary creation of a fluid balance error by altering the correct flow in the circuit of the different machines. Subsequently, the time for alarm occurrence and the threshold value for fluid balance error was evaluated. The alarm was overridden and the overall fluid error allowed by the machine was evaluated. Each machine was tested in conditions of different dialysate/filtrate flow rates and in different simulated treatment modalities. RESULTS: Fluid balance errors can be easily avoided not only by a correct and careful adherence to the protocols of use of the current CRRT machines, but also by the compliance to prescriptions and programmed controls during therapy. Most importantly, if an alarm appears on the machine, one can try to override it without major problems; major problems may occur when multiple override commands are operated without identifying the problem and solving it adequately. CONCLUSION: Machines seem to be designed with adequate safety features and accurate alarm systems. However, features and alarms can be manipulated by operators creating the opportunity for serious error. Physicians and nurses involved in prescription and delivery of CRRT should have precise protocols and defined procedures in relation to machine alarms to prevent major clinical problems. PMID- 16211527 TI - Hemofiltration for cytokine-driven illnesses: the mediator delivery hypothesis. AB - Hemofiltration is evolving as an adjunctive therapy for sepsis and other forms of systemic inflammation. Designed as a substitute for lost renal function, it is sometimes employed prior to the onset of renal failure to facilitate the nonspecific clearance of pro-inflammatory mediators. Prevailing theories suggest that hemofiltration attenuates the immune response when a threshold amount of excess cytokine is removed at the semi-permeable membrane. In this article we introduce an alternative hypothesis, in which hemofiltration exerts its effect by reinvigorating lymphatic flow and function. Crystalloid "replacement" solution, as much as 48 to 72 liters daily, is infused to restore intravascular volume lost through production of ultrafiltrate. Partial redistribution into interstitium and lymph mobilizes inflammatory mediators and other proteins, cellular byproducts, excessive ground matrix, fragments of apoptotic cells and free DNA. These substances are then metabolized, scavenged or cleared at multiple sites, including the reticuloendothelial system, liver, kidney, erythrocyte, and hemofilter. PMID- 16211528 TI - ECMO: back where we started, again. PMID- 16211529 TI - Facial changes in adult uremic patients on chronic dialysis: possible role of hyperparathyroidism. AB - BACKGROUND: Uremic patients on regular dialytic treatment (RDT) are often affected by a complex metabolic syndrome leading to osteodystrophy. Bone changes are primarily due to high bone turnover, often combined with a mineralization defect leading to increased bone fractures and bone deformities. Although rarely considered, the craniofacial skeleton represents one of the peculiar targets of this complex metabolic disease whose more dramatic pattern is a form of leontiasis ossea. This complication, although described, has never been evaluated in depth nor quantitatively assessed. In order to assess facial deformities in uremic conditions and to understand the possible relation with hyperparathyroidism, we undertook a quantitative evaluation of soft facial structures in a cohort of uremic patients undergoing RDT. METHODS: The three dimensional coordinates of 50 soft-tissue facial landmarks were obtained by an electromagnetic digitizer in 10 male and 10 female patients with chronic renal insufficiency aged 53-81 years, and in 34 healthy individuals of the same age, ethnicity and sex. Uremic patients were enrolled according to hyperparathyroid status (PTH < 300 pg/mL and PTH > 500 pg/mL). From the landmarks, facial distances, angles and volumes were calculated according to a geometrical face model. RESULTS: Overall, the uremic patients had significantly larger facial volumes than the reference subjects. The effect was particularly evident in the facial middle third (maxilla), leading to an inversion of the mandibular maxillary ratio. Facial dimensions were increased in all three spatial directions: width (skull base, mandible, nose), length (nose, mandible), and depth (mid face, mandible). The larger maxilla was accompanied by a tendency to more prominent lips (reduced interlabial angle). Some of the facial modifications (nose, lips, mandible) were significantly related to the clinical characteristics of the patients (age, duration of renal insufficiency and PTH levels). CONCLUSIONS: This report, the first in the literature, shows that facial structures of uremic patients are enlarged in comparison with matched normal subjects and that increased bone turnover could be responsible--at least in part- for facial bone changes. PMID- 16211530 TI - Effectiveness of aluminum hydroxide timing administration in relation to meals in controlling hyperphosphatemia in dialysis patients. AB - Phosphate binder compounds contribute to the control of hyperphosphatemia in hemodialysis (HD) patients. However, the most effective schedule of administration of phosphate binders in relation to meals is not well documented. We examined the effectiveness of aluminum hydroxide intake as the sole phosphate binder in relation to meals. Eighty-five patients on regular HD (45 male, 40 female), age 21-72 years, with a duration of 6-216 months HD participated in the study. In all patients, phosphate binders were discontinued for a one month period. Thereafter, and according to the protocol, all patients were advised to take aluminum hydroxide [Al(OH)3 ] 30 min before, during and 30 min after meals for 3 periods of one month each, in a random order. One month washout period preceded the periods of Al(OH)3 ingestion. When Al(OH)3 was administered 30 min prior to the meals, serum phosphate decreased by 7.0% (0.59 mg/dL), while when administrated with or 30 min after meals, it decreased statistically significantly by 28.5% (2.08 mg/dL), and 16% (1.29 mg/dL) respectively. Our results suggest that the efficacy of Al(OH)3 to bind phosphate salts and thus to prevent the hyperphosphatemia in HD patients is higher when this drug is taken with meals. PMID- 16211531 TI - Pharmacokinetics of intraperitoneal cefazolin and ceftazidime coadministered to CAPD patients. AB - PURPOSE: Guidelines for empiric treatment of PD-related peritonitis published in 2000 recommend concurrent intraperitoneal (IP) cefazolin and ceftazidime. The pharmacokinetics (PK) of these agents combined have not been studied. This study was designed to determine the PK of combined IP cefazolin and ceftazidime in CAPD patients. DESIGN: Prospective PK study in seven non-infected CAPD patients. PROCEDURES: Patients had a peritoneal equilibration test (PET), then received one IP dose of cefazolin and ceftazidime (15 mg/kg each) co-administered over a 4 hour dwell, then performed three CAPD exchanges over the next 16 hours. Serum and dialysate samples collected over the 20-hour study period were assayed for drug concentrations by HPLC. OUTCOME MEASURES: PK parameters. STATISTICAL METHODS: Correlations were tested between PET and PK parameters using the Pearson-product correlation coefficient. MAIN FINDINGS: Serum cefazolin and ceftazidime levels exceeded the minimum inhibitory concentrations for susceptible organisms (8 mg/L) throughout the 20 hour study period. Mean cefazolin and ceftazidime PK parameters included: bioavailability, 71% and 63%; elimination rate constant, 0.031 and 0.045 h -1 ; total clearance, 5.8 and 16.0 ml/min; peritoneal clearance, 1.6 and 3.9 ml/min; renal clearance, 2.3 and 3.9 ml/min, respectively. Predictive equations suggest that 1000 mg IP of cefazolin and of ceftazidime every 24 hours would produce average steady-state trough serum cefazolin and ceftazidime concentrations of 70 +/- 52 mg/L and 17 +/- 7 mg/L, respectively. There was no correlation between PET and PK parameters. CONCLUSIONS: Co-administration did not adversely affect the PK of either agent. IP cefazolin and ceftazidime (15 mg/kg) produced adequate serum and dialysate concentrations in CAPD patients for 20 hours. PK predictions suggest that most patients would achieve adequate cefazolin and ceftazidime concentrations with 1000 mg IP once-daily. Anuric patients and those with significant residual renal function may require a more individualized approach. PMID- 16211532 TI - Hybrid test bench for evaluation of any device related to mechanical cardiac assistance. AB - Hydraulic mock circulatory systems have low flexibility to allow tests of different cardiovascular devices and low precision when a reference model must be reproduced. In this paper a new bench is described. It combines the computer model of the environment in which the device will operate and the electro hydraulic interfaces by which device and computer are connected. A models library provided with basic functions allows implementing many layouts of the bench, which in turn depend both on the device properties and the desired experiment. In case of an apical LVAD evaluation, the bench can reproduce right and left ventricles, pulmonary and systemic circulations, inlet and outlet LVAD cannulas. An interface forces the instantaneous calculated flow at the VAD input and feeds back the measured pressure to the computer; another interface works in a similar but complementary- way at the VAD output. The paper focuses on the operating principle of the electro hydraulic interfaces which represent a relevant component of the bench, on the RT-Linux-based software architecture, on the models of the basic elements of the bench. A patent is under preparation. At the moment, only a portion of the bench has been developed. It consists of a piston cylinder mechanism, which mimics the elastance-based mechanism of a natural ventricle, and a hydraulic circuit representing the arterial load according to a modified windkessel model and the venous return according to the Guyton's model. The pump is driven by a real-time simulation of the cardiovascular system. This preliminary layout allowed testing the piston-cylinder mechanism, its control, and the software. This electro-hydraulic interface has been used to reproduce a pulsatile pump working in different modes. The hybrid model approach can support the development of new cardiac assist devices from their computer model to their manufacture. PMID- 16211533 TI - Engrafting periodontal fibroblasts with new 3-dimensional polylactide foams. AB - In clinical periodontics, "tissue engineering" techniques have been developed to guide the regenerative process following periodontal disease. We explored a new shaped biomaterial in order to promote cellular adhesion, proliferation and migration of periodontal cells. Granules of poly-D, L-lactide were foamed in specially designed moulds by a controlled gas-loading process. Explant cultures of periodontal tissue were seeded at different densities on the 3-dimensional scaffolds following analysis of cytotoxicity, cell proliferation and differentiation. The moulding procedure led to porous structures with predetermined tubes for cellular locomotion. Cells adhered to and populated the material's surface and inner cavities while retaining fibroblastic phenotype. An optimal ratio between cellular proliferation and mortality rate was achieved at a seeding density of > 10 6 cells/cm 3 scaffold. We designed modified polylactide scaffolds capable of acting as a stent and a cell carrier matrix. The foaming process is easy, cheap and suitable for commercial production. PMID- 16211534 TI - PMEA polymer-coated PVC tubing maintains anti-thrombogenic properties during in vitro whole blood circulation. AB - Poly(2-methoxyethylacrylate) (PMEA) is a new coating material that appears to reduce protein and platelet adsorption. However, the exact performance of PMEA coated circuit remains to be revealed in well-controlled experiments. Therefore, we compared its hemocompatibility with covalent-bound heparin-, and non-coated circuits during 6 hours of in vitro circulation, using donor blood from six volunteers. In our model, simple tubing circuits containing one-way ball valve were placed on the rotary table, which moved alternatively to generate pulsatile recirculation of heparinized human blood inside the tubing. Using this model, we expected fine assessment of the material surface, because we could reduce blood damage by avoiding air and a blood pump. Moreover, the small capacity of circuit allowed us to compare three kinds of circuits using a single unit of donor blood, eliminating effects by possible variations between blood donors. The anti thrombin capacity of the PMEA-coated circuits was maintained even after six hours blood circulation, whereas surface thrombin generation increased markedly after use in non-coated circuits (P<0.05). Deposition of fibrin onto PMEA circuits was reduced more than 30% compared with heparin and non-coated circuits (P<0.05). However, the increase of plasma Factor XIIa was similar in all circuits. Increase of CD11b expression on circulating leukocytes and of plasma C3a was ameliorated in the heparin- and PMEA-coated circuits (P<0.05). PMEA-coated circuits appear to maintain their anti-thrombogenicity during use, otherwise PMEA-coated and heparin coated circuits showed a similar character in hemocompatibility. This long standing anti-thrombogenicity might be attributable to less adsorption of activated blood components onto the surface. PMID- 16211535 TI - Shock wave therapy as an innovative technology in skeletal disorders: study on transmembrane current in stimulated osteoblast-like cells. AB - Extracorporeal shock wave treatment (ESWT) is successfully used in various musculoskeletal disorders and pathologies. Despite the increasing use of this kind of therapy, some aspects of its mechanism of action are still unclear. In vitro bone cell behavior under ESWT were previously investigated by the present author and MG63 osteoblast-like cells showed an enhancement in proliferation and in the osteoblast differentiation after therapy with a low-energy flux density. The aim of the present study was to evaluate the effect of ESWT on the permeabilization of cell membrane. We characterized physiological changes in the MG63 associated with ESWT generated by an ESW device and patch clamp recording was performed to study ion channels. Experiments were carried out using the whole cell recording configuration of the patch-clamp technique and the ionic current measurements were performed on cell samples of ESW treated and control groups. The patch-clamp technique showed the effect of ESWT on the amplitude of transmembrane currents. The treatment with ESW enhanced the transmembrane current as well the voltage dependence of Ca-activated and K channels that mediate these currents: the differences between treated cells and control at 80mV were over 1000 pA (p<0.05). These modifications of ion channels activity positively influence cell proliferation (MTT test, p<0.0001) without interfering with the normal synthesis activity of stimulated osteoblasts. PMID- 16211536 TI - Comparative analysis of frozen and acellularized vascular xenografts. AB - BACKGROUND: We implanted frozen and acellularized porcine xenograft vessels as small-diameter arterial grafts in goats and comparatively analyzed the explanted grafts by gross observation and by light microscopy at predetermined periods. MATERIALS AND METHODS: Porcine carotid arteries were harvested and immediately stored within a tissue preservation solution at -70 degrees C in a freezer designated for frozen xenograft vessels. The acellularized xenograft vessels were prepared with NaCl-SDS solution and stored frozen until use. One pair of porcine xenograft vessels were used to compare the frozen and acellularized grafts in the bilateral carotid arteries in one goat. The grafts were implanted for one, 3, and 6 months in three animals. Periodic ultrasonographic examinations were performed during the observation period. Explanted grafts were analyzed by gross observation, and by light microscopy. RESULTS: All animals survived the experimental procedure without specific problems. Ultrasonographic examinations showed excellent patency in all grafts during the observation period. Gross observations revealed nonthrombotic patent smooth lumens. Microscopic examinations of the explanted grafts showed satisfactory cellular reconstruction to the 6-month stage. Although more inflammatory responses were observed in the early phase of implantation of frozen xenografts than of acellularized xenografts, there was no evidence of significant rejection of the frozen xenografts. CONCLUSION: These findings suggest that porcine vessel xenografts, regardless of them being acellularized or simply frozen xenografts, can be acceptably implanted in goats as a form of small-diameter vascular graft. PMID- 16211538 TI - GROMACS: fast, flexible, and free. AB - This article describes the software suite GROMACS (Groningen MAchine for Chemical Simulation) that was developed at the University of Groningen, The Netherlands, in the early 1990s. The software, written in ANSI C, originates from a parallel hardware project, and is well suited for parallelization on processor clusters. By careful optimization of neighbor searching and of inner loop performance, GROMACS is a very fast program for molecular dynamics simulation. It does not have a force field of its own, but is compatible with GROMOS, OPLS, AMBER, and ENCAD force fields. In addition, it can handle polarizable shell models and flexible constraints. The program is versatile, as force routines can be added by the user, tabulated functions can be specified, and analyses can be easily customized. Nonequilibrium dynamics and free energy determinations are incorporated. Interfaces with popular quantum-chemical packages (MOPAC, GAMES-UK, GAUSSIAN) are provided to perform mixed MM/QM simulations. The package includes about 100 utility and analysis programs. GROMACS is in the public domain and distributed (with source code and documentation) under the GNU General Public License. It is maintained by a group of developers from the Universities of Groningen, Uppsala, and Stockholm, and the Max Planck Institute for Polymer Research in Mainz. Its Web site is http://www.gromacs.org. PMID- 16211540 TI - The GROMOS software for biomolecular simulation: GROMOS05. AB - We present the latest version of the Groningen Molecular Simulation program package, GROMOS05. It has been developed for the dynamical modelling of (bio)molecules using the methods of molecular dynamics, stochastic dynamics, and energy minimization. An overview of GROMOS05 is given, highlighting features not present in the last major release, GROMOS96. The organization of the program package is outlined and the included analysis package GROMOS++ is described. Finally, some applications illustrating the various available functionalities are presented. PMID- 16211539 TI - Integrated Modeling Program, Applied Chemical Theory (IMPACT). AB - We provide an overview of the IMPACT molecular mechanics program with an emphasis on recent developments and a description of its current functionality. With respect to core molecular mechanics technologies we include a status report for the fixed charge and polarizable force fields that can be used with the program and illustrate how the force fields, when used together with new atom typing and parameter assignment modules, have greatly expanded the coverage of organic compounds and medicinally relevant ligands. As we discuss in this review, explicit solvent simulations have been used to guide our design of implicit solvent models based on the generalized Born framework and a novel nonpolar estimator that have recently been incorporated into the program. With IMPACT it is possible to use several different advanced conformational sampling algorithms based on combining features of molecular dynamics and Monte Carlo simulations. The program includes two specialized molecular mechanics modules: Glide, a high throughput docking program, and QSite, a mixed quantum mechanics/molecular mechanics module. These modules employ the IMPACT infrastructure as a starting point for the construction of the protein model and assignment of molecular mechanics parameters, but have then been developed to meet specialized objectives with respect to sampling and the energy function. PMID- 16211542 TI - Glycine-based polymeric surfactants with varied polar head group: I. synthesis, characterization, and application in micellar electrokinetic chromatography. AB - The monomers and polymers of four anionic amide type sodium undecenoxy carbonyl glycinate (SUCG) surfactants and four anionic carbamate type sodium undecenoyl glycinate (SUG) surfactants with 1-, 2-, 3-, and 4-glycine unit as head group were synthesized and characterized. The CMC and aggregation number (A) for all eight surfactants were determined using fluorescence spectroscopy. In addition, the CMC values of these surfactants were also projected by surface tension and CE. The CMC of the monomers decreases with increases in the size of glycine head groups and correlates well when the fluorescence method was compared to CE. The A number increases and partial specific volume (V) decreases with increase in size of the head group of both monomers and polymers. However, A and V are always lower for the polymers than the corresponding monomers. The electrophoretic and chromatographic parameters of micelle polymers of SUG and SUCG were also examined. The coefficient of EOF increases with the increase in size of the head group but the electrophoretic mobility decreases which results in a decrease in the elution range. The retention data suggest that the selectivity differences among the mono-, di-, and tripeptide derivatives of poly-SUCG surfactants are relatively higher compared to the derivatives of poly-SUG series. PMID- 16211541 TI - Electrochemiluminescence of dipicolinic acid (DPA) and (bpy)(2)Ru(DPA)(+) (bpy = 2,2'-bipyridine). AB - The spectroscopic and electrochemiluminescence (ECL) properties of dipicolinic acid (DPA), (bpy)(2)Ru(2+) (bpy = 2,2'-bipyridine) and the species formed when DPA and (bpy)(2)Ru(2+) [abbreviated to (bpy)(2)Ru(DPA)(+)] are allowed to react are reported. The UV-Vis absorption maxima for (bpy)(2)Ru(2+) and (bpy)(2)Ru(DPA)(+) are 493 and 475 nm, respectively, indicating the in situ formation of a complex between DPA and (bpy)(2)Ru(2+). DPA, (bpy)(2)Ru(2+) and (bpy)(2)Ru(DPA)(+) display ECL upon oxidation in the presence of the oxidative reductive co-reactant tri-n-propylamine (TPrA). The ECL of (bpy)(2)Ru(DPA)(+) is at least two-fold higher than either of the parent species. An ECL spectrum of (bpy)(2)Ru(DPA)(+) displays a peak maximum 40 nm red-shifted from the photoluminescence peak maximum, suggesting that the excited state formed electrochemically is different from that formed spectroscopically. PMID- 16211543 TI - Modulation of osteopontin in proteinuria-induced renal interstitial fibrosis. AB - Proteinuria is associated with macrophage-dependent interstitial fibrosis (IF). Osteopontin (OPN), a macrophage chemoattractant, may be involved in the transition of proteinuria to IF but protective properties have also been reported. To elucidate whether OPN may be involved in the proteinuria-induced cascade of tubulointerstitial damage, renal expression of OPN was studied during the development of proteinuria-induced renal damage and during anti-proteinuric intervention with ACE inhibition (ACEi). First, the temporal relationships between proteinuria, interstitial OPN induction, and IF in adriamycin nephrosis (AN), a model of chronic proteinuria-induced renal damage, were studied. Second, the effect of anti-proteinuric treatment on OPN expression was investigated. The time course of OPN induction and markers of renal damage was studied in rats with unilateral AN at 6-week intervals until week 30. In a second study, a renal biopsy was taken 6 weeks after induction of bilateral AN; subsequently, rats were treated with ACEi until termination (week 12). In unilateral AN, proteinuria developed gradually and stabilized at week 10. In proteinuric kidneys, OPN expression was induced from week 12 onwards. Simultaneously, a progressive increase in interstitial macrophages, alpha-smooth muscle actin (alpha-SMA), collagen type III, and focal glomerulosclerosis (FGS) was observed. In bilateral AN, ACEi reduced proteinuria and OPN protein and stabilized fibrosis. In untreated animals, OPN mRNA increased, with stable OPN protein and fibrosis and increased FGS. Thus, in AN, development of proteinuria is followed by up regulation of OPN along with markers of renal damage. The up-regulation of OPN is reversible by anti-proteinuric treatment without a corresponding reduction in fibrosis. Whereas these data are consistent with a role for OPN in the cascade of transition from proteinuria to fibrosis, intervention with ACEi showed that reduction of OPN does not attenuate established fibrosis. PMID- 16211544 TI - Targeted chemotherapy with cytotoxic bombesin analogue AN-215 can overcome chemoresistance in experimental renal cell carcinomas. AB - BACKGROUND: Multidrug resistance (MDR) mediated by membrane transporters, such as P-glycoprotein (MDR-1) and MDR-associated protein (MRP), remains a challenge in the therapy of renal cell carcinoma (RCC). Chemotherapy targeted to hormone receptors may provide a new approach to overcome chemoresistance. The cytotoxic analogue of bombesin/gastrin-releasing peptide (GRP), AN-215, consists of a superactive derivative of doxorubicin, AN-201, which is linked to a bombesin analogue carrier: RC-3094. METHODS: The authors examined the expression of bombesin/GRP receptors in 3 human RCC cell lines (A-498, ACHN. and 786-0) by using reverse-transcriptase-polymerase chain reaction (RT-PCR) analysis and radioligand-binding assays. They also evaluated the effects of AN-215 and its cytotoxic radical AN-201 in the same RCC models in vivo, and they studied the effects of AN-215 and AN-201 on the expression levels of MDR-1 and subtype 1 of MRP (MRP-1) by using real-time PCR. RESULTS: A N-215 significantly (P < 0.05) inhibited the growth of A-498, ACHN, and 786-0 RCC xenografted into nude mice by 59.2-67.6%, whereas the cytotoxic radical AN-201 alone had no significant antitumor effects. The efficacy of AN-215 was independent of the expression patterns of MDR-1 and MRP-1 in these RCC cell lines. The induction of MDR-1 by AN 215 was similar (Experiment 2) or weaker (Experiment 1) compared with AN-201. Both AN-215 and AN-201 caused only a minor induction of MRP-1. CONCLUSIONS: The current findings indicated that targeted chemotherapy with cytotoxic bombesin/GRP analogue AN-215 can inhibit the growth of RCC, providing a new treatment modality for patients with advanced RCC. PMID- 16211545 TI - Zeta-chain associated protein 70 and CD38 combined predict the time to first treatment in patients with chronic lymphocytic leukemia. AB - BACKGROUND: Zeta-chain associated protein (ZAP)-70 has been proposed as a surrogate marker for immunoglobulin heavy-chain variable region (IgVH) mutation in chronic lymphocytic leukemia (CLL), but it is still not clear whether it is an independent prognostic factor. METHODS: The authors evaluated ZAP-70 expression by flow cytometry in 201 untreated patients and correlated ZAP-70 levels with CD38 expression, genetic abnormalities detected by fluorescence in situ hybridization (FISH), and the time from diagnosis to first treatment. RESULTS: Fifty-seven patients (28%) were positive for ZAP-70 (> or = 20%). Positive ZAP-70 status was associated with advanced disease stage, atypical morphology, CD38 positive status, trisomy 12, del(6q), or no detectable abnormalities; negative ZAP-70 status was correlated with del(13q) as a sole abnormality. The treatment free interval (TFI) was 17.7 months for ZAP-70-positive patients and 44.6 months for ZAP-70-negative patients (P < 0.001). Multivariate analysis in 117 patients identified advanced stage, CD38 > or = 7%, and the absence of del(13q) as a sole abnormality as independent factors for short TFI. Excluding FISH, ZAP-70 status acquired independent prognostic value along with CD38 status. The authors proposed a risk model that combines ZAP-70 and CD38 to identify patients who are likely to progress. When both markers were positive, the TFI was 12 months; when both were negative, the median TFI was 54 months; a median TFI of 26 months was observed in patients who had discordant results (P < 0.00001). CONCLUSIONS: The current findings suggested that both ZAP-70 and CD38 should be tested prospectively in all patients with early-stage CLL. PMID- 16211546 TI - Anaplastic large cell lymphoma treated with a leukemia-like therapy: report of the Italian Association of Pediatric Hematology and Oncology (AIEOP) LNH-92 protocol. AB - BACKGROUND: Childhood anaplastic large cell lymphoma (ALCL) is a well defined entity with a rather poor prognosis. Different approaches have been adopted in the treatment of ALCL in various cooperative trials, including short high-dose intensive therapy and leukemia-like protocols. In the early 1990s, the Italian Association of Pediatric Hematology and Oncology (AIEOP) initiated a multicenter trial for the treatment of ALCL based on a modified LSA2-L2 protocol. METHODS: Thirty-four consecutive eligible children with newly diagnosed ALCL were enrolled in the AIEOP LNH-92 protocol. Treatment was comprised of an induction of remission phase, followed by consolidation and maintenance for a total duration of 24 months, independently of disease stage. RESULTS: Thirty of 34 patients (88%) achieved complete disease remission and 8 patients experienced disease recurrence. With a median follow-up of 8.4 years, the probabilities of survival and event-free survival were 85% (range, 79-91%) and 65% (range, 57-73%), respectively. Therapy was well tolerated and hematologic toxicity was the most frequent toxicity. CONCLUSIONS: The leukemia-like protocol AIEOP LNH-92 was found to be an effective treatment for childhood ALCL. Its long duration may be beneficial to specific patient subgroups, but optimal treatment duration in ALCL remains to be elucidated. PMID- 16211547 TI - Differences in the quality of breast cancer care among vulnerable populations. AB - BACKGROUND: It is unknown whether differences in the quality of breast cancer care among women from racial and ethnic minority groups, the elderly, and rural areas have changed over time across the continuum of care. METHODS: The linked Surveillance, Epidemiology, and End Results-Medicare database identified 22,701 women ages 66-79 years diagnosed with early stage breast cancer from 1992-1999. Multiple breast cancer processes of care were measured, including breast conserving surgery, radiation therapy, documentation of estrogen receptor status, surveillance mammography, and a combined measure of "adequate care". RESULTS: African-American and Hispanic women were significantly less likely to receive adequate care than White women in unadjusted comparisons (54.7% and 58.0% vs. 68.4% for African-American and Hispanic vs. White women) and adjusted comparisons (adjusted odds ratio [AOR] 0.67; 95% confidence interval [95% CI] 0.59-0.76, and AOR 0.77; 95% CI 0.66-0.90 for African-American and Hispanic women, respectively). The proportion of Asian/Pacific Islander women receiving adequate care was similar to White women. When considering only women diagnosed with breast cancer from 1997-1999, African-American women remained less likely than White women to receive adequate care (AOR 0.63; 95% CI 0.50-0.79). Women ages 75 79 years were less likely to receive adequate care compared with women ages 66-69 years (AOR 0.74; 95% CI 0.69-0.80), and women from rural (vs. metropolitan) areas were less likely to receive adequate care (AOR 0.81; 95% CI 0.73-0.89). CONCLUSIONS: The quality of breast cancer care is lower among vulnerable populations across the continuum of care, and many of these differences have not improved in more recent years. PMID- 16211548 TI - Microstructure formation and property of chitosan-poly(acrylic acid) nanoparticles prepared by macromolecular complex. AB - We report here a study on the microstructure formation process of polymeric nanoparticles based on polyelectrolyte complexes. When polyanion poly(acrylic acid) (PAA) was dropped into polycation chitosan (CS) solution, CS-PAA nanoparticles with diverse microstructure would be formed under different experimental conditions. The microstructure of CS-PAA nanoparticles changed from solid spherical nanoparticles to core-shell separative ones and turned back to solid spherical ones with the variation of preparation conditions. The influence of molecular weight of CS and PAA, shell cross-linking, dropping temperature on the size, stability and morphology of CS-PAA nanoparticles were also studied. The nanoparticle size was affected by the molecular weight of CS and PAA, the ratio of amino group to carboxyl group (na/nc) and the incubation temperature as well. The shell-cross-linking provides a means to stabilize these nanoparticles. These nanoparticles can encapsulate plasmid DNA very well, which makes them have great potential in gene delivery. PMID- 16211549 TI - In-vitro hemocompatibility evaluation of a thermoplastic polyurethane membrane with surface-immobilized water-soluble chitosan and heparin. AB - The surface of a thermoplastic polyurethane (TPU) membrane was treated with low temperature plasma (LTP) and was then grafted with poly(acrylic acid) (PAA), followed by the grafting of water-soluble chitosan (WSC) and heparin (HEP). The surface was characterized with static contact-angle and X-ray photoelectron spectroscopy (XPS). The results showed that the surface densities of peroxides and PAA reached a maximum when treated with LTP for 90 s. A higher pH of the reacting solution led to higher graft densities of WSC and HEP. After WSC and HEP grafting, the hydrophilicity of the TPU membrane was increased. The adsorption of proteins on HEP-grafted TPU membranes was effectively curtailed. In addition, HEP grafting also reduced platelet adhesion, elevated thrombin inactivation, and prolonged the blood coagulation time. According to the L929 fibroblast cell growth inhibition index, the HEP-grafted TPU membranes exhibited non cytotoxicity. Overall results demonstrated that the HEP immobilization could not only improve the hydrophilicity but also the hemocompatibility of the TPU membrane, while maintaining the ascendant biocompatibility. PMID- 16211550 TI - Interpolymer complex between hydroxypropyl cellulose and maleic acid-styrene copolymer: phase behavior of semi-dilute solutions. AB - The phase behavior of a water/hydroxypropyl cellulose/maleic acid-styrene copolymer (H2O/HPC/MAc-S) system was investigated in the semi-dilute range by turbidimetry, rheology, and optical microscopy. The two polymers under investigation form interpolymer complexes via hydrogen bonding. In the case of a total polymer concentration of cpol = 5 mg . mL(-1) a second phase segregates upon heating the homogeneous ternary system. By applying a constant shear rate (gamma = 50 s(-1)) the phase separation temperature of the system is 10-15 degrees C lower than for an unsheared one. For cpol = 10 mg . mL(-1) phase separation has already occurred at room temperature when the two binary polymer solutions are mixed. The distribution of the partners among the coexisting phases was examined by FT-IR spectroscopy. The stoichiometry of the interpolymeric complex (IPC) was estimated to be HPC/MAc-S = 40:60 (w/w) independent of cpol. PMID- 16211552 TI - Convalescent whole blood, plasma and serum in the prophylaxis of measles: JAMA, 12 April, 1926; 1180-1187. PMID- 16211551 TI - Preparation and characterisation of thermoresponsive poly[(N-isopropylacrylamide co-acrylamide-co-(hydroxyethyl acrylate)] microspheres as a matrix for the pulsed release of drugs. AB - Despite the large number of publications and patents concerning pH/thermoresponsive polymers, few data are available concerning the preparation of thermoresponsive cross-linked microspheres from preformed polymers. Therefore, N-isopropylacrylamide-co-acrylamide-co-(2-hydroxyethyl acrylate) copolymers were obtained as a new thermoresponsive material with a lower critical solution temperature (LCST) around 36 degrees C, in phosphate buffer at pH 7.4, and with a cross-linkable OH group in their structure. The LCST value was determined both by UV spectroscopy and microcalorimetric analysis. These copolymers were solubilised in acidified aqueous solution below their LCST, dispersed in mineral oil, and transformed into stable microspheres by cross-linking with glutaraldehyde. The thermoresponsive microspheres were characterised by optical and scanning electron microscopy, degree of swelling, and water retention. The pore dimensions of the microspheres and the retention volumes of some drugs and typical compounds were evaluated at different temperatures by liquid chromatography. Indomethacin, as a model drug, was included in the microspheres by the solvent evaporation method. Finally, the influence of temperature and of temperature cycling on drug release was investigated. PMID- 16211553 TI - Genetic variant characterization in intron 4 of the surfactant protein B gene. AB - Genetic variants in intron 4 of the surfactant protein B gene SFTPB have been associated with pulmonary morbidity in newborn infants and adults. Genetic variant discovery in intron 4 requires high fidelity polymerase amplification due to a variable number of intermotif dinucleotide repeats and reliable characterization of alleles genetically distinct due to insertion, deletion, or both of 11 conserved sequence motifs. To optimize genetic variant discovery, we selected a high fidelity polymerase enzyme by replicate amplification and compared automated sequencing with agarose gel electrophoresis for all variant alleles (N=68) in a cohort of Missouri infants with (N=187) and without (N=53) respiratory distress. We identified and characterized six new alleles based on sequence motifs and two pairs of variant alleles with similar mobilities by agarose gel electrophoresis but distinct motif sequences. We confirmed uniformity of invariant alleles by sequencing (N=77). Logistic regression using race and intron 4 genotype for infants > or = 35 weeks gestation suggested that the invariant allele was independently associated with increased risk of respiratory distress (OR for the invariant allele 2.7, 95% CI 1.0-7.2). Reliable characterization of genetic variants in intron 4 requires both a high fidelity polymerase and sequencing of variant alleles. PMID- 16211556 TI - Nine mutations including three novel mutations among Russian patients with acute intermittent porphyria. AB - Acute intermittent porphyria (AIP) is a metabolic disease due to a partial deficiency of hydroxymethylbilane synthase (HMBS) in heme biosynthesis. Direct sequencing of genomic DNA samples of 11 unrelated Russian AIP patients, 32 of their relatives and 50 healthy controls from northwestern Russia including Saint Petersburg revealed nine mutations in the HMBS gene. Three novel mutations, c.825+5G>C, c.825+3_825+6del, and c.770T>C, resulted in varying amounts of abnormal transcripts, r.822_825del and [r.770U>C, r.652_771del, r.613_771del]. Six mutations, c.77G>A (p.R26H), c.517C>T (p.R173W), c.583C>T (p.R195C), c.673C>T (p.R225X), c.739T>C (p.C247R), and c.748G>C (p.E250A), have previously been identified in AIP patients from Western and other Eastern European populations. All mutations expressed in COS-1 cells demonstrated low residual activities (0.1 1%). The majority of the mutations were family-specific and also confirmed allelic heterogeneity among Russian AIP patients. The diversity of mutations may reflect the old international history of Saint Petersburg and immigration of people from other parts of Europe. PMID- 16211554 TI - Molecular characterization and cancer risk associated with BRCA1 and BRCA2 splice site variants identified in multiple-case breast cancer families. AB - Genetic screening of women from multiple-case breast cancer families and other research-based endeavors have identified an extensive collection of germline variations of BRCA1 and BRCA2 that can be classified as deleterious and have clinical relevance. For some variants, such as those in the conserved intronic splice site regions which are highly likely to alter splicing, it is not possible to classify them based on the identified DNA sequence variation alone. We studied 11 multiple-case breast cancer families carrying seven distinct splice site region genetic alterations in BRCA1 or BRCA2 (BRCA1, c.IVS6-2delA, c.IVS9-2A>C, c.IVS4-1G>T, c.IVS20+1G>A and BRCA2, c.IVS17-1G>C, c.IVS20+1G>A, c.IVS7-1G>A) and applied SpliceSiteFinder to predict possible changes in efficiency of splice donor and acceptor sites, characterized the transcripts, and estimated the average age-specific cumulative risk (penetrance) using a modified segregation analysis. SpliceSiteFinder predicted and we identified transcipts that illustrated that all variants caused exon skipping, and all but two led to frameshifts. The risks of breast cancer to age 70 yrs, averaged over all variants, over BRCA1 variants alone, and over BRCA2 variants alone, were 73% (95% confidence interval 47-93), 64% (95%CI 28-96) and 79% (95%CI 48-98) respectively (all P<0.0001). Therefore five of these seven consensus splice site variants of BRCA1 and BRCA2 produce a transcript similar to that of other previously described deleterious exonic variants and are associated with similar high lifetime risks. PMID- 16211557 TI - The first missense alteration in the MCPH1 gene causes autosomal recessive microcephaly with an extremely mild cellular and clinical phenotype. AB - Autosomal recessive primary microcephaly (MCPH) is a rare neurodevelopmental disorder characterized by mental retardation and congenital microcephaly with a head circumference at least 4 SD below age and sex means, in the absence of other significant malformations or neurological deficits. Truncating alterations in the MCPH1 gene have previously been shown to exhibit a distinct cellular phenotype, with a high proportion of prophase-like cells (>10%) due to premature chromosome condensation in early G2- and delayed decondensation in early G1-phase of the cell cycle. We report here the first patient with a homozygous substitution of a highly conserved threonine residue by an arginine (c.80C>G, Thr27Arg) localized in the N-terminal BRCT domain of MCPH1. The cellular and clinical phenotype of this patient is much less pronounced than that of previously described patients with truncating alterations in the MCPH1 gene. Firstly, the fraction of prophase like cells accounts for just 3-4% of the cell population. Secondly, clinically, he has only a very mild mental retardation with predominantly delayed motor skills but normal verbal IQ attainment. Additionally, head circumference was less severely affected, being -2.4 SD at birth and -3 SD at the age of six years. This justifies reconsideration and widening of the clinical phenotype definition of MCPH1. PMID- 16211558 TI - Novel mutations of the PCSK9 gene cause variable phenotype of autosomal dominant hypercholesterolemia. AB - Autosomal dominant hypercholesterolemia (ADH) is a frequent (1/500) monogenic inherited disorder characterized by isolated elevation of LDL leading to premature cardiovascular disease. ADH is known to result from mutations at two main loci: LDLR (encoding the low density lipoprotein receptor), and APOB (encoding apolipoprotein B100), its natural ligand. We previously demonstrated that ADH is also caused by mutations of the PCSK9 (proprotein convertase subtilisin/kexin type 9) gene that encodes Narc-1 (neural apoptosis-regulated convertase 1). However, the role of this novel disease locus as a cause of hypercholesterolemia remains unclear. In the present study, we analysed the PCSK9 coding region and intronic junctions in 130 adult or pediatric patients with ADH, previously found as being non LDLR/non APOB mutation carriers. Four novel heterozygous missense variations were found: c.654A>T (p.R218S), c.1070G>A (p.R357H), c.1405C>T (p.R469W), and c.1327G>A (p.A443T). All mutations were absent in 340 normolipidemic controls. Except for the A443T, all mutations are nonconservative and modify a highly conserved residue. Segregation with hypercholesterolemia is incomplete in one pedigree. Type and severity of hyperlipidemia and of cardiovascular disease could vary among subjects from the same family. Finally, the proband carrying the R357H mutation exhibited very high plasma cholesterol during pregnancy, whereas the proband carrying the p.R469W mutation exhibited a severe phenotype of hypercholesterolemia in combination with a LDLR mutation resulting from a frameshift at residue F382 (1209delC). These observations suggest that variations in PCSK9 are a rare cause of non LDLR/non APOB ADH (approximately 2.3%) and that additional environmental or genetic factors may contribute to the phenotype caused by PCSK9 missense mutations in humans. PMID- 16211559 TI - Alterations in N-methyl-D-aspartate receptor sensitivity and magnesium blockade occur early in development in the R6/2 mouse model of Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder that affects primarily the striatum and cerebral cortex. A search for the factors that increase the vulnerability of striatal neurons will lead to a better understanding of the pathological cascades of this disease. A current hypothesis for neurodegeneration of striatal medium-sized spiny neurons in HD is an alteration in N-methyl-D-aspartate (NMDA) receptor function. In the present study we examined electrophysiological properties of NMDA receptors in the R6/2 transgenic mouse model. These animals express exon 1 of the human HD gene and present an overt behavioral phenotype at about 5 weeks of age. Whole-cell voltage clamp recordings from acutely dissociated striatal neurons were obtained from three different age groups of transgenic mice (15, 21, and 40 days old) and their littermate controls (WT). In transgenic animals, two groups of neurons were found with respect to NMDA and Mg2+ sensitivity. One group of R6/2 cells displayed responses similar to those of WT, whereas the other showed increased responses to NMDA and decreased Mg2+ sensitivity. These cells were encountered in all age groups. The abnormal sensitivity to NMDA and Mg2+ indicates that NMDA receptor alterations occur very early in development and suggest the presence of constitutively abnormal NMDA receptors. These alterations may contribute to an enhancement of NMDA responses at hyperpolarized membrane potentials that may be a key factor in striatal neuronal dysfunction. PMID- 16211560 TI - Regional estimates of hair cells and supporting cells in the human crista ampullaris. AB - Regional estimates of type I and type II vestibular hair cells (HC) and supporting cell (SC) numbers were obtained from the horizontal crista ampullaris by using design-based stereology in human. Cristae were microdissected from temporal bones obtained post-mortem (N=16, age range 26-98 years). Three groups were made according to age: group 1, n=5, ages between 26 and 67 years, average age 51 years; group 2, n=4, average age 84 years; and group 3, n=7, average age 94 years. For group 1, the average total HC number was 8,005+/-214, corresponding to 4,119+/-107 type I HC, 3,886+/-117 type II HC, and 10,274+/-224 SC. The type I:type II HC ratio was 1.06+/-0.01, and HC density was 0.80 cells/100 microm2. For group 2, the average total HC number was 7,074+/-489, corresponding to 3,733+/-212 type I HC, 3,341+/-314 type II HC, and 9,321+/-858 SC. The type I:II HC ratio was 1.12+/-0.06, and HC density was 0.75 cells/100 microm2. For group 3, the average HC number was 6,009+/-327, corresponding to 3,380+/-223 type I HC, 2,628+/-235 type II HC, and 10,185+/-182 SC. The type I:II HC ratio was 1.34+/ 0.10, and HC density was 0.63 cells/100 microm2. A significant decline in type I, type II, and total HC number and density was found in groups 2 and 3, with individuals exceeding the average human life span. PMID- 16211561 TI - Human and mouse microglia express connexin36, and functional gap junctions are formed between rodent microglia and neurons. AB - Microglia, the tissue macrophages of the central nervous system (CNS), intimately interact with neurons physically and through soluble factors that can affect microglial activation state and neuronal survival and physiology. We report here a new mechanism of interaction between these cells, provided by the formation of gap junctions composed of connexin (Cx) 36. Among eight Cxs tested, expression of Cx36 mRNA and protein was found in microglial cultures prepared from human and mouse, and Cx45 mRNA was found in mouse microglial cultures. Electrophysiological measurements found coupling between one-third of human or mouse microglial pairs that averaged below 30 pico-Siemens and displayed electrical properties consistent with Cx36 gap junctions. Importantly, similar frequency of low strength electrical coupling was also obtained between microglia and neurons in cocultures prepared from neocortical or hippocampal rodent tissue. Lucifer yellow dye coupling between neurons and microglia was observed in 4% of pairs tested, consistent with the low strength and incidence of electrical coupling. Cx36 expression level and/or the degree of coupling between microglia did not significantly change in the presence of activating agents, including lipopolysaccharide, granulocyte-macrophage colony-stimulating factor, interferon gamma, and tumor necrosis factor-alpha, except for some reduction of Cx36 protein when exposed to the latter two agents. Our findings that intercellular coupling occurs between neuronal and microglial populations through Cx36 gap junctions have potentially important implications for normal neural physiology and microglial responses in neuronopathology in the mammalian CNS. PMID- 16211562 TI - Sulforaphane enhances aquaporin-4 expression and decreases cerebral edema following traumatic brain injury. AB - Brain edema, the infiltration and accumulation of excess fluid causing an increase in brain tissue volume, often leads to a rise in intracranial pressure and is a key contributor to the morbidity and mortality associated with traumatic brain injury (TBI). The cellular and molecular mechanisms contributing to the development/resolution of TBI-associated brain edema are poorly understood. Aquaporin-4 (AQP4) water channel is expressed at high levels in brain astrocytes, and the bidirectional transport of water through these channels is critical for the maintenance of brain water homeostasis. By using a rodent injury model, we show that TBI decreased AQP4 level in the injury core and modestly increased it in the penumbra region surrounding the core. Postinjury administration of sulforaphane (SUL), an isothiocyanate present in abundance in cruciferous vegetables such as broccoli, attenuated AQP4 loss in the injury core and further increased AQP4 levels in the penumbra region compared with injured animals receiving vehicle. These increases in AQP4 levels were accompanied by a significant reduction in brain edema (assessed by percentage water content) at 3 days postinjury. These findings suggest that the reduction of brain edema in response to SUL administration could be due, in part, to water clearance by AQP4 from the injured brain. PMID- 16211564 TI - Microhardness and fracture toughness of dental materials by indentation method. AB - The main objective of this study was to measure the fracture toughness of the human teeth enamel using the microindentation technique and to compare the results calculated from the equations developed for Palmqvist and radial-median cracks. Vickers microhardness measurements of dental ceramic (alumina) and human teeth were performed using indentation fracture method. The fracture toughness of enamel was calculated using different equations reported in the literature. Vickers microhardness of the sintered alumina specimen (98.8% theoretical density) was measured to be 14.92 GPa under 9.8 N indentation load. Three equations based on the radial-median cracks were found to be applicable for the fracture toughness determination of the enamel. Results show that indentation fracture method is adequate to measure microhardness and fracture toughness of dental materials. However the calculation of fracture toughness depended on the nature of the cracks and also on the location of the indentation. Therefore, it is necessary to identify the crack profile and to select the appropriate equation for accurate fracture toughness values. PMID- 16211563 TI - Morphine activates Arc expression in the mouse striatum and in mouse neuroblastoma Neuro2A MOR1A cells expressing mu-opioid receptors. AB - Activity-regulated cytoskeleton-associated protein (Arc) is an effector immediate early gene product implicated in long-term potentiation and other forms of neuroplasticity. Earlier studies demonstrated Arc induction in discrete brain regions by several psychoactive substances, including drugs of abuse. In the present experiments, the influence of morphine on Arc expression was assessed by quantitative reverse transcription real-time PCR and Western blotting in vivo in the mouse striatum/nucleus accumbens and, in vitro, in the mouse Neuro2A MOR1A cell line, expressing mu-opioid receptor. An acute administration of morphine produced a marked increase in Arc mRNA and protein level in the mouse striatum/nucleus accumbens complex. After prolonged opiate treatment, tolerance to the stimulatory effect of morphine on Arc expression developed. No changes in the striatal Arc mRNA levels were observed during spontaneous or opioid antagonist-precipitated morphine withdrawal. In Neuro2A MOR1A cells, acute, but not prolonged, morphine treatment elevated Arc mRNA level by activation of mu opioid receptor. This was accompanied by a corresponding increase in Arc protein level. Inhibition experiments revealed that morphine induced Arc expression in Neuro2A MOR1A cells via intracellular signaling pathways involving mitogen activated protein (MAP) kinases and protein kinase C. These results lend further support to the notion that stimulation of opioid receptors may exert an activating influence on some intracellular pathways and leads to induction of immediate early genes. They also demonstrate that Arc is induced in the brain in vivo after morphine administration and thus may play a role in neuroadaptations produced by the drug. PMID- 16211565 TI - Meniscal replacement in dogs. Tissue regeneration in two different materials with similar properties. AB - In earlier studies, meniscal replacement with a porous polymer implant led to regeneration of neo-meniscal tissue. To evaluate the influence of the chemical properties on the tissue regeneration in the implant, in the present study, the meniscus in the dog's knee was replaced with either an aromatic 4,4 diphenylmethanediisocyanate based polyesterurethane implant (Estane) (n = 6) or with an aliphatic 1,4-butanediisocyanate based polyesterurethane implant (PCLPU) (n = 6). After 6 months, the knee joints were resected and the tissue behavior in the two different prostheses was evaluated microscopically. In both prostheses, a meniscus-like distribution of the tissue phenotype was found with collagen type I in the peripheral fibrous zones and collagen type II in the central, more cartilaginous zones. The compression-stress behavior of the implant-tissue construct remained in between the stiffness of the polymer material and that of the native meniscus. The PCLPU implant seemed to provoke less synovial tissue reaction. After meniscectomy solely, in 5 out of 6 cases, a meniscus-like regenerate was formed. Furthermore, the articular cartilage degeneration after placing a PCLPU implant did also not exceed the degeneration after the Estane implant or after meniscectomy. The differences between these two implants did not seem to influence the tissue regeneration in the implant. However, PCLPU seemed to evoke less tissue reaction and, therefore, is thought to be less or even nontoxic as compared with the Estane implant. Therefore, for studies in the future, the authors prefer the PCLPU prostheses for replacement of the meniscus. PMID- 16211566 TI - Biological response to a new composite polymer augmentation device used for cruciate ligament reconstruction. AB - A resorbable composite augmentation cord braided of poly(L-lactide) and poly(L lactide-co-glycolide) fibers was designed for the temporary protection of repaired cruciate ligaments. This study examined the biocompatibility of the new device and the influence of augmentation duration on ligament healing in a sheep model. The anterior cruciate ligament (ACL) was cut close to the femoral insertion and reinserted with sutures. The repaired ACLs were augmented with the slowly degrading new composite cord and alternatively with a faster degrading polydioxanone cord (PDS). A tendon graft group (gold standard) served as control. Histological evaluation and biomechanical testing were performed after 6 months. The composite cord showed no signs of degradation, whereas the PDS was intra articularly resorbed. Both devices showed only minor foreign body reactions, proving their good biocompatibility. However, 9 of 11 composite cords had ruptured too early because of fatigue at the bone tunnel entrances. All operated knees were less stable than the nonoperated collateral joints. Knees equipped with the composite cord showed the largest anterior instabilities, whereas the PDS-augmented group exhibited in some cases knee instabilities comparable with that of the tendon group. A positive effect of a longer mechanical protection by a slowly degrading augmentation could not yet be shown. The fatigue strength of the device still needs improvement. PMID- 16211567 TI - Gene expression by human monocytes from peripheral blood in response to exposure to metals. AB - With increasing life expectancy and active lifestyles, the longevity of arthroplasties has become an important problem in orthopaedic surgery and will remain so until novel approaches to joint preservation have been developed. The sensitivity of the recipient to the metal alloys may be one of the factors limiting the lifespan of implants. In the present study, the response of human monocytes from peripheral blood to an exposure to metal ions was investigated, using the method of real-time polymerase chain reaction (PCR)-based low-density arrays. Upon stimulation with bivalent (Co2+ and Ni2+) and trivalent (Ti3+) cations and with the calcium antagonist LaCl3, the strength of the elicited monocytic response was in the order of Co2+ > or = Ni2+ > Ti3+ > or = LaCl3. The transcriptional regulation of the majority of genes affected by the exposure of monocytes to Co2+ and Ni2+ was similar. Some genes critically involved in the processes of inflammation and bone resorption, however, were found to be differentially regulated by these bivalent cations. The data demonstrate that monocytic gene expression is adapted in response to metal ions and that this response is, in part, specific for the individual metals. It is suggested that metal alloys used in arthroplasties may affect the extent of inflammation and bone resorption in the peri-implant tissues in dependence of their chemical composition. PMID- 16211568 TI - Structural analysis of dextran-based hydrogels obtained chemoenzymatically. AB - This work reports the results of structural analysis in novel dextran-acrylate (dexT70-VA) hydrogels generated chemoenzymatically. Porous structure as well as hydrogel surface and interior morphologies were evaluated by mercury intrusion porosimetry (MIP), nitrogen adsorption (NA), and scanning electron microscopy (SEM) analyses, as a function of the degree of substitution (DS), and initial water content used in the preparation of the hydrogel. MIP analysis showed that the overall networks were clearly macroporous with pore sizes ranging from 0.065 to 10 microm. As expected, the average pore size decreased as DS increased and as initial water content decreased. Moreover, the porosity values ranged from 75 up 90%, which shows that these hydrogels present an interconnected pore structure. Nitrogen adsorption analyses showed that the specific surface area of dexT70-VA hydrogels increased either by increasing the DS or by decreasing the initial water content of the hydrogel. SEM results revealed that the surface of hydrogels with lower DS presented either a porous structure or a polymeric "skin" covering the pores, whereas hydrogels with higher DS were totally porous. Furthermore, the interior morphology varied according to the DS and the initial water content of the hydrogels. Finally, the average pore size was also determined from the swelling of hydrogel using a theoretical model developed by Flory-Rehner. The comparison of the SEM and MIP results with the ones obtained by the equilibrium swelling theory of Flory-Rehner shows that this approach highly underestimates the average pore size. PMID- 16211569 TI - An in vivo study of tricalcium phosphate and glutaraldehyde crosslinking gelatin conduits in peripheral nerve repair. AB - In order to modulate the mechanical properties of gelatin, we previously developed a biodegradable composite composed by tricalcium phosphate and glutaraldehyde crosslinking gelatin (GTG) feasible for surgical manipulation. In this study, we evaluated the in vivo applications of GTG conduit for peripheral nerve repair. The effect of sciatic nerve reconstruction was compared between resorbable permeable GTG conduits and durable impermeable silicone tubes. Traditional methods of assessing nerve recovery following peripheral nerve repair including histomorphometric and electrophysiologic features were conducted in our study. In addition, autotomy score and sciatic function index (SFI) in walking tract analysis were used as additional parameters for assessing the return of nerve function. Twenty-four weeks after sciatic nerve repair, the GTG conduits were harvested. Microscopically, regeneration of nerves was observed in the cross section at the mid portion of all implanted GTG conduits. The cross-sectional area of regenerated nerve of the GTG group was significant larger than that of the silicone group. In the compound muscle action potentials (CMAP), the mean recovery index of CMAP amplitude was 0.24 +/- 0.02 for the silicone group, 0.41 +/- 0.07 for the GTG group. The mean SFI increased with time in the GTG group during the evaluation period until 24 weeks. Walking tract analysis showed a higher SFI score in the GTG group at both 12 and 24 weeks. The difference reached a significant level at 24 weeks. Thus, the histomorphometric, electrophysiologic, and functional assessments demonstrate that GTG can be a candidate for peripheral nerve repair. PMID- 16211570 TI - Frequency effect in fretting wear of Co-28Cr-6Mo versus Ti-6Al-4V implant alloys. AB - A piezo-electrically driven fretting testing device has been constructed and fretting release and release rates have been determined with highest accuracy, using a radiotracer technique. First results on the fretting release and release rate of titanium alloy fretting pads against cobalt-chrome alloy fatigue specimens are reported. The frequency dependency of fretting release has been determined between 1 and 8 Hz and shows higher release rates for low frequencies, thus indicating that accelerated testing of materials and components of artificial joints must be analyzed extremely carefully. The present experiments under simple conditions present a base-line study for step-wise applying more complex and realistic testing conditions and for using radiotracer methods to quantify fretting release in simulated testing of artificial hip- and knee prostheses. PMID- 16211571 TI - Enhancing the mechanical integrity of the implant-bone interface with BoneWelding technology: determination of quasi-static interfacial strength and fatigue resistance. AB - The BoneWelding technology is an innovative bonding method, which offers new alternatives in the treatment of fractures and other degenerative disorders of the musculoskeletal system. The BoneWelding process employs ultrasonic energy to liquefy a polymeric interface between orthopaedic implants and the host bone. Polymer penetrates the pores of the surrounding bone and, following a rapid solidification, forms a strong and uniform bond between implant and bone. Biomechanical testing was performed to determine the quasi-static push-out strength and fatigue performance of 3.5-mm-diameter polymeric dowels bonded to a bone surrogate material (Sawbones solid and cellular polyurethane foam) using the BoneWelding process. Fatigue tests were conducted over 100,000 cycles of 20-100 N loading. Mechanical test results were compared with those obtained with a comparably-sized, commercial metallic fracture fixation screw. Tests in surrogate bone material of varying density demonstrated significantly superior mechanical performance of the bonded dowels in comparison to conventional bone screws (p < 0.01), with holding strengths approaching 700 N. Even in extremely porous host material, the performance of the bonded dowels was equivalent to that of the bone screws. For both cellular and solid bone analog materials, failure always occurred within the bone analog material surrounding and distant to the implant; the infiltrated interface was stronger than the surrounding bone analog material. No significant decrease in interfacial strength was observed following conditioning in a physiological saline solution for a period of 1 month prior to testing. Ultrasonically inserted implants migrated, on average, less than 20 microm over, and interfacial stiffness remained constant the full duration of fatigue testing. With further refinement, the BoneWelding technology may offer a quicker, simpler, and more effective method for achieving strong fixation and primary stability for fracture fixation or other orthopaedic and dental implant applications. PMID- 16211572 TI - Injectable biomaterials for incontinence and vesico-ureteral reflux: current status and future promise. AB - Many injectable biomaterials have been produced as bulking agents for compression of urethral sphincter or ureteral orifice for treating adult stress incontinence or vesico-ureteral reflux in pediatrics. The agents being developed include glutaraldehyde crosslinked collagen, dextranomer/hyaluronic acid copolymer, pyrolytic carbon-coated zirconium oxide beads, polydimethyl-siloxane microparticles, polytetrafluoroethylene paste, autologous fats, autologous chondrocytes, and others. Though less invasive nature of these agents has gained their popularity as a quick solution of the disease symptoms, most of such treatments fail to produce good long-term efficacy. The failure is likely caused by the rapid degradation of material implants and the lack of tissue regeneration/integration properties. We thus believe that a good injectable biomaterial for incontinence should possess the following two properties: (1) to resist degradation and to reside in the implantation sites for a long period of time or (2) to enhance tissue regeneration and to establish permanent periurethral or subureteric tissue. Here we report some recent results for supporting this hypothesis. PMID- 16211573 TI - Micro-tensile bond strength of sound and caries-affected primary tooth dentin measured with original designed jig. AB - To measure the micro-tensile bond strength (MTBS) on sound and caries-affected primary tooth dentin using an original designed jig that was developed for making symmetric and uniformly-sized specimens for the micro-tensile strength testing. The MTBS of dumbbell-shaped specimens and matchstick-shaped specimens were measured. Dumbbell-shaped specimens: The occlusal surfaces of 15 sound primary molars were ground with a water-cooled air turbine using a diamond bur. The adhesive systems and resin-based composites were Clearfil SE Bond and Clearfil AP X (SE group) and Single Bond and Z250 (SB group). Matchstick-shaped specimens: Sixteen primary molars (5 sound and 11 carious) were used. The occlusal surfaces of sound teeth were ground with a water-cooled air turbine using a diamond bur. Infected dentin was determined by a caries detecting dye solution and removed with a round-shaped steel bur under water-cooling and with a hand instrument. SE was applied. Data was statistically analyzed using ANOVA and Fisher's PLSD at p < 0.05. The MTBSs (MPa) for the dumbbell-shaped specimens were 8.9 +/- 7.0 in the SE group and 10.3 +/- 5.8 in the SB group and that for the matchstick-shaped specimens were 9.0 +/- 6.3 in the sound-SE group and 9.2 +/- 5.8 in the carious SE group. No significant difference was observed among them. PMID- 16211574 TI - Hemocompatibility of low-friction boron-carbon-nitrogen containing coatings. AB - Mechanical heart valves are exposed to extreme mechanical demands, which require a surface showing not only nonhaemostatic properties, but also wear resistance and low friction. As alternative to different forms of amorphous carbon (a-C), so called diamond-like carbon (DLC), the suitability of boron carbonitride (BCN) coatings is tested here for hemocompatible coatings. They have similar mechanical properties like a-C surfaces, but superior chemical stability at ferrous substrates or counterparts. BCN films with different nitrogen content were compared with hydrogenated a-C films regarding their mechanical properties, surface energy, adsorption of albumin and fibrinogen, blood platelet adherence, and activation of the contact system of the clotting cascade and kinin system. Similar mechanical properties and biological response have been found in the BCN films with respect to a-C, indicating the potential of these coatings for biomedical applications. The increase in the crystallinity and tribological properties of the BCN samples with a higher incorporation of N was also followed by a lower protein adsorption and low activation of the contact system, but an increased adherence of thrombocytes. PMID- 16211575 TI - Deciphering the human nucleolar proteome. AB - Nucleoli are plurifunctional nuclear domains involved in the regulation of several major cellular processes such as ribosome biogenesis, the biogenesis of non-ribosomal ribonucleoprotein complexes, cell cycle, and cellular aging. Until recently, the protein content of nucleoli was poorly described. Several proteomic analyses have been undertaken to discover the molecular bases of the biological roles fulfilled by nucleoli. These studies have led to the identification of more than 700 proteins. Extensive bibliographic and bioinformatic analyses allowed the classification of the identified proteins into functional groups and suggested potential functions of 150 human proteins previously uncharacterized. The combination of improvements in mass spectrometry technologies, the characterization of protein complexes, and data mining will assist in furthering our understanding of the role of nucleoli in different physiological and pathological cell states. PMID- 16211576 TI - Characterization of the promoter for the alpha3 integrin gene in various tumor cell lines: roles of the Ets- and Sp-family of transcription factors. AB - The alpha3beta1 integrin is an adhesion receptor for extracellular matrix proteins, including laminin isoforms, and plays crucial roles in the organization of epithelial and endothelial tissues. The aberrant expression of this adhesion molecule on tumor cells is associated with their invasive and metastatic potentials. In the present study, we analyzed the elements essential for alpha3 integrin gene expression in various tumor cell lines with different tissue origins by luciferase assay. An approximately 0.3 kb fragment of the 5'-flanking region of the mouse alpha3 integrin gene (-260/+84, relative to the major transcription start site) showed strong promoter activity in all six examined tumor cell lines. However, we found that these cell lines could be divided into two groups according to the level of dependency on the putative Ets-transcription factor binding motif located at -133. This motif was previously shown to be crucial for alpha3 integrin expression in MKN1 gastric carcinoma cells. The gene expression in one group of cell lines was upregulated mainly by the Ets motif, whereas that in the other group was less dependent on the Ets motif. We then postulated that additional regulatory elements were responsible for the expression of alpha3 integrin, and found that a GC-rich motif at -69 was another important element. An electrophoretic mobility shift assay using specific antibodies and a Western blot analysis of nuclear proteins revealed that the Sp3 transcription factor bound to this GC-rich motif. These results suggest that the Sp3 and Ets transcription factors cooperatively regulate alpha3 integrin gene expression and that the contribution of each element depends on the type of tumor cells. PMID- 16211577 TI - Absence of SPARC in lens epithelial cells results in altered adhesion and extracellular matrix production in vitro. AB - The matricellular protein SPARC (also known as osteonectin and BM-40) is expressed abundantly in lens epithelium. That SPARC-null mice exhibit early cataractogenesis, indicates a role for SPARC in the maintenance of lens transparency. Comparison of cultured wild-type and SPARC-null lens epithelial cells revealed significant changes in adhesion to different substrates. SPARC null lens cells displayed enhanced attachment and spreading, focal adhesion formation, and resistance to trypsin detachment in comparison to wild-type cells. In the absence of SPARC, there was increased deposition of the ECM protein laminin-1 (LN-1). Proteins associated with focal adhesions were increased in SPARC-null versus wild-type lens cells: levels of alpha6-integrin heterodimers, talin, and paxillin phosphorylated on tyrosine were enhanced significantly, as was the association of beta1-integrin with talin and paxillin. Restoration of the wild-type phenotype in SPARC-null cultures was accomplished through genetic rescue by stable transfection of SPARC cDNA. Our findings indicate that SPARC is counter-adhesive for murine lens epithelial cells and demonstrate that multiple factors contribute to this activity. We also identify SPARC as a modulator of LN 1 secretion and deposition by these cells, an activity important in epithelial cell-ECM interactions in the ocular lens. PMID- 16211578 TI - Aryl hydrocarbon receptor, cell cycle regulation, toxicity, and tumorigenesis. AB - Most effects of exposure to halogenated and polycyclic aromatic hydrocarbons are mediated by the aryl hydrocarbon receptor (AHR). It has long been recognized that the AHR is a ligand-activated transcription factor that plays a central role in the induction of drug-metabolizing enzymes and hence in xenobiotic detoxification. Of late, it has become evident that outside this well characterized role, the AHR also functions as a modulator of cellular signaling pathways. In this Prospect, we discuss the involvement of the AHR in pathways critical to cell cycle regulation, mitogen-activated protein kinase cascades, immediate-early gene induction, and the functions of the RB protein. Ultimately, the toxicity of AHR xenobiotic ligands may be intrinsically connected with the perturbation of these pathways and depend on the many critical signaling pathways and effectors with which the AHR itself interacts. PMID- 16211579 TI - Solution structures of the reduced and Cu(I) bound forms of the first metal binding sequence of ATP7A associated with Menkes disease. AB - The coding sequence for the first N-terminal copper binding motif of the human Menkes disease protein (MNK1; residues 2-79) was synthesized, cloned, and expressed in bacteria for biochemical and structural studies. MNK1 adopts the betaalphabetabetaalphabeta fold common to all the metal binding sequences (MBS) found in other metal transport systems (e.g., the yeast copper chaperone for superoxide dismutase CCS, the yeast copper chaperone ATX1 bound to Hg(II), and most recently Cu(I), the bacterial copper binding protein, CopZ, and the bacterial Hg(II) binding protein MerP), although substantial differences were found in the metal binding loop. Similar to ATX1, MNK1 binds Cu(I) in a distorted linear bicoordinate geometry. As with MerP, MNK1 has a high affinity for both Hg(II) and Cu(I), although it displays a marked preference for Cu(I). In addition, we found that F71 is a key residue in the compact folding of MNK1, and its mutation to alanine results in an unfolded structure. The homologous residue in MerP has also been mutated with similar results. Finally, to understand the relationship between protein folding and metal affinity and specificity, we expressed a chimeric MBS with the MNK1 protein carrying the binding motif of MerP (CAAC-MNK1); this chimeric protein showed differences in structure and the dynamics of the binding site that may account for metal specificity. PMID- 16211580 TI - Interleukin-1beta induces osteopontin expression in pulmonary fibroblasts. AB - Osteopontin is a multifunctional matricellular protein identified as one of the most upregulated genes in pulmonary fibrosis. Experimental animal models have identified early pro-fibrotic cytokines as essential to the pathogenesis of inflammation-induced pulmonary fibrosis. However, the principal sources of osteopontin in the fibroproliferative lung, and the factors responsible for its induction, have not been fully defined. We isolated primary rat lung fibroblasts in culture to examine the expression and regulation of lung fibroblast-derived osteopontin. Our results demonstrate a potent and dramatic increase in osteopontin expression induced by interleukin-1beta (IL-1beta), whereas tumor necrosis factor-alpha, transforming growth factor-beta, and angiotensin II had minimal effect. Stimulation with IL-1beta resulted in the secretion of soluble osteopontin protein. We found that osteopontin expression by IL-1beta was regulated via signaling primarily through the mitogen-activated protein kinase member ERK1/2, partially by p38 MAPK, but not at all by JNK. Finally, the mechanism of IL-1beta increase in osteopontin mRNA requires de novo transcription and translation. In conclusion, we find that osteopontin is expressed by primary lung fibroblasts and is potently upregulated by the early inflammatory and pro fibrotic cytokine IL-1beta. Activated fibroblasts may be a significant source of osteopontin production during lung fibrogenesis. PMID- 16211581 TI - Study of hepatocyte differentiation using embryonic stem cells. AB - The liver has many crucial functions including metabolizing dietary molecules, detoxifying compounds, and storing glycogen. The hepatocytes, comprising most of the liver organ, progressively modify their gene expression profile during the fetal development according to their roles in the different phases of development. Embryonic stem (ES) cells serve as a major tool in understanding liver development. These cells may also serve as a source of hepatic cells for cellular therapy. In this review, we aim to summarize the research that has been performed in the field of hepatocyte differentiation from mouse and human ES cells. We discuss the various methodologies for the differentiation of ES cells towards hepatic cells using either spontaneous or directed differentiation protocols. Although many protocols for differentiating ES cells to hepatic cells have been developed, the analysis of their status is not trivial and can lead to various conclusions. Hence, we discuss the issues of analyzing hepatocytes by means of the specificity of the markers for hepatocytes and the status of the cells as fetal or adult hepatocytes. PMID- 16211582 TI - Inner nuclear membrane and signal transduction. AB - Recent research has shown that the inner nuclear membrane is a site for regulation of signal transduction from the plasma membrane to the nucleus. This has coincided with discoveries showing that mutations in extrinsic and intrinsic inner nuclear membrane proteins cause a variety of inherited diseases. In most instances, the mechanisms by which mutations in inner nuclear membrane proteins cause disease are not understood. In at least one case, however, an alteration in signal transduction appears to underlie disease pathogenesis. PMID- 16211583 TI - Induction of chemokines and chemokine receptors CCR2b and CCR4 in authentic human osteoclasts differentiated with RANKL and osteoclast like cells differentiated by MCP-1 and RANTES. AB - Chemokines MCP-1 and RANTES are induced when authentic bone resorbing human osteoclasts differentiate from monocyte precursors in vitro. In addition, MCP-1 and RANTES can stimulate the differentiation of cells with the visual appearance of osteoclasts, being multinuclear and positive for tartrate resistance acid phosphatase (TRAP +). We show here that MIP1alpha is also potently induced by RANKL during human osteoclast differentiation and that this chemokine also induces the formation of TRAP + multinucleated cells in the absence of RANKL. MIP1alpha was able to overcome the potent inhibition of GM-CSF on osteoclast differentiation, permitting the cells to pass through to TRAP + multinuclear cells, however these were unable to form resorption pits. Chemokine receptors CCR2b and CCR4 were potently induced by RANKL (12.6- and 49-fold, P = 4.0 x 10( 7) and 4.0 x 10(-8), respectively), while CCR1 and CCR5 were not regulated. Chemokine treatment in the absence of RANKL also induced MCP-1, RANTES and MIP1alpha. Unexpectedly, treatment with MCP-1 in the absence of RANKL resulted in 458-fold induction of CCR4 (P = 1.0 x 10(-10)), while RANTES treatment resulted in twofold repression (P = 1.0 x 10(-4)). Since CCR2b and CCR4 are MCP-1 receptors, these data support the existence of an MCP-1 autocrine loop in human osteoclasts differentiated using RANKL. PMID- 16211584 TI - Neurofilaments can undergo axonal transport and cytoskeletal incorporation in a discontinuous manner. AB - Neurofilaments (NFs) are thought to provide structural support for axons. Some NFs exhibit an extended residence time along axons, the nature of which remains unclear. In prior studies in NB2a/d1 cells, hypophosphorylated NFs were demonstrated to be dispersed throughout the axon and to undergo relatively rapid axonal transport, while extensively phosphorylated NFs organized into a "bundle" localized along the center of the axon. It was not conclusively determined whether bundled NFs underwent transport or instead underwent turnover via exchange with transporting individual NFs. Herein, using transfection with multiple constructs and regional photobleaching, we demonstrate that bundled NFs undergo relatively slow transport as well as exchange with surrounding individual NFs. We also demonstrate that newly synthesized NFs disperse nonhomogenously throughout axonal neurites and perikarya. These findings provide a mechanism by which some NFs exhibit extended residence time within axons, which lessens the metabolic burden of cytoskeletal turnover. PMID- 16211585 TI - Rho family GTPases are activated during HGF-stimulated prostate cancer-cell scattering. AB - An important process in embryogenesis and cancer-cell metastasis is the conversion of epithelial cells to a migratory phenotype, a phenomenon known as epithelial-mesenchymal transition (E-MT). To achieve E-MT, cells dissociate from neighbouring cells and adopt a migratory morphology. This transition requires remodelling of their cell shape and substratum adhesions; activities that require extensive reorganisation of the actin cytoskeleton. Hepatocyte growth factor (HGF)-induced scattering of Madin Darby canine kidney (MDCK) cells is a routinely used model of E-MT, in which actin cytoskeletal rearrangement is known to be dependent on Rho family GTPases. We have developed a novel model of HGF-induced E MT using the human prostate cancer cell line, DU145. This model overcomes the limitation of using a canine cell line and facilitates the study of E-MT in human cancer. We demonstrate for the first time the scattering response of individual DU145 cells to HGF in real time and have characterised changes in actin cytoskeletal organisation and cell adhesions as these cells respond to HGF. HGF induced scattering of DU145 cells is dependent on the activity of Rho family GTPases, and using this model, we are able to demonstrate for the first time that endogenous Cdc42 is activated downstream of HGF. Furthermore we have also shown that the response of DU145 cells to HGF is dependent on a phosphatidylinositide 3 kinase pathway. PMID- 16211586 TI - Signaling molecularly imprinted polymers: molecular recognition-based sensing materials. AB - Molecular imprinting is a template polymerization technique that can easily provide synthetic polymers capable of molecular recognition for given target molecules. In addition to their highly specific recognition ability, we are attempting to introduce signaling functions to molecularly imprinted polymers, enabling them to respond according to specific binding events. Some of our work regarding such signaling molecularly imprinted polymers is presented here, including molecularly imprinted polymers that induce spectral shifts of target compounds because of binding. Such compounds include hydrogen-bonding-based fluorescent imprinted polymers and metalloporphyrin-based signaling molecularly imprinted polymers. PMID- 16211587 TI - Electronically-coupled MM quadruply-bonded complexes of molybdenum and tungsten. AB - The reaction of M2(O2CBu(t))4 (M = Mo, W) with a dicarboxylic acid in toluene yields compounds of general formula [M2]-O2C-X-CO2-[M2] ([M2] = M2(O2CBu(t))3; X = conjugated spacer). The M2 units are electronically coupled via interactions between the M2 delta and dicarboxylate pi* orbitals, and the magnitude of this coupling is revealed by electronic structure calculations and spectroscopic data. These compounds show intense metal to ligand charge transfer (MLCT) absorptions in the visible region of the electronic spectrum that are temperature and solvent dependent. Evidence of electronic coupling is seen in their cyclic voltammograms, which show two successive one-electron oxidations. The extent of electronic coupling in the mixed valence radical cations [M2]-O2C-X-CO2-[M2]+, generated by oxidation with one equivalent of AgPF6 or FeCp2PF6, is evaluated by EPR and UV vis-NIR spectroscopic data, and delocalized behavior is observed in compounds with W2 units separated by up to 13.6 angstroms. The simplicity of the frontier M2 orbital interactions with the bridge pi orbitals provides a convenient system with which to study electron transfer in mixed valence systems, as compared to the extensively studied, but more complicated, dinuclear t(2g)6/t(2g)5 mixed valence compounds. Oligomeric and polymeric compounds incorporating M2 units have also been synthesized, having general formula [M2(O2CR)2(O2C-Thio-CO2)]n (Thio = n-hexyl substituted ter- and quinque-thiophenes). They can be deposited as thin films by spin coating, and show photoluminescence and electroluminescence. These metallo-polythiophenes show potential for application in electronic materials. ( PMID- 16211589 TI - Nigral degeneration and striatal dopaminergic dysfunction in idiopathic and Parkin-linked Parkinson's disease. AB - We have used MR segmented inversion recovery ratio imaging (SIRRIM) of the substantia nigra pars compacta to detect and correlate nigral signal change in idiopathic Parkinson's disease (PD) and parkin patients with striatal (18)F-dopa uptake. Nine PD patients, nine parkin patients, and eight control subjects were studied with a combination of MR inversion recovery sequences sensitive to nigral cell loss. Blinded independent observer rating and quantified nigral signal analysis were performed on all subjects. Striatal regions of interest were defined on T(1)-weighted MRI co-registered to (18)F-dopa positron emission tomography. On blinded observer rating of the SIRRIM dorsal and ventral nigral images, 25% (2/8) of control subjects, 44% (4/9) of PD patients, and 67% (6/9) of parkin patients were classified as abnormal. Quantified total nigral signal intensities were reduced to a greater extent in the parkin compared to PD patients. There was a greater predilection for signal reduction in the ventral nigral slice of the PD compared to the parkin patient group, who showed a more uniform involvement. All PD and parkin patients were discriminated from controls on the basis of caudate and putamen (18)F-dopa Ki reductions. Our results suggest that MR segmented inversion recovery ratio imaging shows poor sensitivity for discriminating parkin and idiopathic PD patients from normal controls. Where nigral signal abnormalities were seen, parkin patients manifested generalized nigral cell loss with widespread striatal dopamine terminal dysfunction compared with the lateral nigral targeting seen in PD and selective loss of putamen (18)F dopa uptake. PMID- 16211590 TI - Psychogenic parkinsonism: a combination of clinical, electrophysiological, and [(123)I]-FP-CIT SPECT scan explorations improves diagnostic accuracy. AB - We evaluated the concordance between independent clinical, electrophysiological, and [(123)I]-FP-CIT SPECT scan explorations as a staged procedure for an accurate diagnosis in 9 patients referred with a diagnosis of suspected psychogenic parkinsonism. Three patients were reclassified as pure psychogenic parkinsonism (PP), 6 with a form of combined psychogenic parkinsonism and Parkinson's disease (PP + PD), and none with pure Parkinson's disease (PD). Electrophysiological recordings showed the characteristics of psychogenic tremor in 5 of 7 patients with tremor. In two of these 5, PD tremor was also recorded. SPECT scan results were abnormal in five of 9 patients. In one case of clinically suspected PP + PD, SPECT scan results were normal. Long-term follow-up supported the final diagnosis of PP (initial clinical misdiagnosis). Electrophysiology contributes to the clinical diagnosis of psychogenic tremor and may help confirm associated organic PD tremor. [(123)I]-FP-CIT SPECT is a robust test to ascertain dopaminergic denervation and increase the confidence of the clinical and electrophysiological diagnosis of associated PD. A combination of clinical, electrophysiological, and [(123)I]-FP-CIT SPECT scan explorations improves diagnostic accuracy in order to distinguish PP from PP + PD. PMID- 16211591 TI - Provisional diagnostic criteria for depression in Parkinson's disease: report of an NINDS/NIMH Work Group. AB - Mood disorders are the most common psychiatric problem associated with Parkinson's disease (PD), and have a negative impact on disability and quality of life. Accurate diagnosis of depressive disturbances in PD is critical and will facilitate the testing and use of new interventions; however, there are no clear diagnostic criteria for depressive disorders in PD. In their current form, strict Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria are difficult to use in PD and require attribution of specific symptoms to PD itself or the depressive syndrome. Additionally, DSM criteria for major depression and dysthymia exclude perhaps half of PD patients with comorbid clinically significant depression. This review summarizes an NIH-sponsored workshop and describes recommended changes to DSM diagnostic criteria for depression for use in PD. Participants also recommended: (1) an inclusive approach to symptom assessment to enhance reliability of ratings in PD and avoid the need to attribute symptoms to a particular cause; (2) the inclusion of subsyndromal depression in clinical research studies of depression of PD; (3) the specification of timing of assessments for PD patients with motor fluctuations; and (4) the use of informants for cognitively impaired patients. The proposed diagnostic criteria are provisional and intended to be defined further and validated but provide a common starting point for clinical research in PD associated depression. PMID- 16211593 TI - Inhibition of catechol-O-methyltransferase contributes to more stable levodopa plasma levels. AB - The short plasma half-life limits the antiparkinsonian efficacy of levodopa/carbidopa (LD/CD). Administration of LD/CD with the catechol-O methyltransferase inhibitor entacapone in one tablet (LCE) may extend plasma half life of LD and thus its effect on motor symptoms in patients with Parkinson's disease (PD). The objectives of this study were to monitor the motor response to a switch from LD/CD to LCE by a simultaneous performance of an instrumental motor test and rating of motor symptoms and to compare the LD plasma behavior between both conditions in terms of stability. Twenty-one treated PD patients received LD/CD and then the identical oral LD dosage of LCE within a standardized setting on 2 consecutive days. Rating better reflected the motor improvement after LD application than the instrumental test. Motor symptoms of PD patients decreased significantly more during the LCE than the LD/CD condition, probably due to significantly higher LD plasma levels and a significantly less pronounced fall of the LD concentrations following the second LD intake. Our study shows a more stable LD plasma behavior during LCE intake and accordingly a better effect on motor symptoms according to rating outcomes and motor test results to a lesser extent. PMID- 16211594 TI - Pergolide versus levodopa monotherapy in early Parkinson's disease patients: The PELMOPET study. AB - Dopamine agonists are used as initial treatment in patients with Parkinson's disease (PD) to reduce incidence and severity of motor complications. This paradigm is based on long-term studies, allowing "rescue" therapy with levodopa. The present strict monotherapy study (PELMOPET, the acronym for the pergolide versus-L-dopa-monotherapy-and-positron-emission-tomography trial) evaluated the efficacy and safety of pergolide versus levodopa without levodopa "rescue" medication. This multicenter, double-blind, randomized, 3-year trial compared pergolide monotherapy (n=148) with levodopa monotherapy (n=146) in dopamine-naive patients with early PD (Hoehn and Yahr stage 1-2.5). Primary efficacy measures were clinical efficacy, severity and time to onset of motor complications, and disease progression. During the 3 years, severity of motor complications was significantly lower and time to onset of dyskinesia was significantly delayed in the group receiving pergolide (3.23 mg/day) compared with those receiving levodopa (504 mg/day). However, time to onset of motor complications was not longer in patients receiving pergolide after 3 years. Symptomatic relief (assessed by Unified Parkinson's Disease Rating Scale [UPDRS], UPDRS II, and III, Clinical Global Impressions [CGI] severity, and CGI and Patient Global Impressions [PGI] improvement) was significantly greater in patients receiving levodopa. Adverse events led to discontinuation of therapy in 17.6% of pergolide patients and 9.6% of levodopa patients. This is the first study comparing strict monotherapy with a dopamine agonist versus levodopa in previously untreated early PD. In principle, both levodopa and a dopamine agonist such as pergolide seem to be suitable options as initial PD therapy. The choice remains with the treating physician based on the different efficacy and adverse event profiles. PMID- 16211595 TI - Cognitive profiles of individual patients with Parkinson's disease and dementia: comparison with dementia with lewy bodies and Alzheimer's disease. AB - We describe the pattern of cognitive profiles within a community-based sample of patients with Parkinson's disease (PD) and dementia (PDD) using cluster analyses, and compare the results with data from patients with Alzheimer's disease (AD) and dementia with Lewy bodies (DLB). Fifty patients with PDD and 39 with AD from Stavanger, Norway, and 62 patients with DLB from San Diego, CA, USA were diagnosed by either standardized clinical procedures or criteria (all PDD and all AD cases) or necropsy (all DLB cases). Four subgroups were identified: two subgroups with a subcortical cognitive profile (one with mild and one with moderate dementia severity), one subgroup with global impairment and severe dementia, and one subgroup with a cortical cognitive profile and moderate dementia. Of the patients with PDD and with DLB, 56% and 55%, respectively, had a subcortical cognitive profile, compared with only 33% of the AD patients. Conversely, 30% of the patients with PDD and 26% of those with DLB had a cortical cognitive profile, compared with 67% of the patients with AD. These findings suggest that in some patients with PDD, frontosubcortical changes are the main contributing factor to dementia, whereas in other patients, cortical and hippocampal changes are more important. PMID- 16211596 TI - Subthalamic nucleus influences spatial orientation in extra-personal space. AB - While the role of frontal and parietal cortex in spatial orientation has been studied extensively, the contribution of the basal ganglia and especially the subthalamic nucleus to spatial orientation remains less clear. Here we use subthalamic nucleus (STN) deep brain stimulation (DBS) in Parkinson's disease (PD) as a reversible model of functional lesioning to evaluate the influence of the STN in extra-personal space orientation. To this end, 12 PD patients were examined 1 year after implantation of DBS electrodes in the STN after overnight withdrawal of L-dopa. Patients were tested in a pseudo-randomized order while both stimulators, the right only, the left only, or no stimulator, were switched on. Patients performed line bisection and a reaction time task responding to stimuli of the middle, the left, and the right extra-personal space. A separate assessment of the right and left hand responding to visual stimuli in each hemispace made it possible to distinguish hemi-spatial and hemi-motor impairments. No asymmetries in space orientation were found when both stimulators were switched OFF, when both stimulators were switched ON, and when only the right stimulator was switched ON. When only the left subthalamic stimulation was switched ON, the reaction times of both hands to visual stimuli in the left extra personal hemispace increased significantly and the line bisection test showed a significant orientation to the right. These results lead to the conclusion that the STN and its cortical projections influence the network involved in visuospatial orientation. These patterns of symptoms of neglect demonstrate the influence of the STN on the attentional system of the nondominant hemisphere. PMID- 16211597 TI - The effect of 6-dimethylaminopurine (6-DMAP) and cycloheximide (CHX) on the development and chromosomal complement of sheep parthenogenetic and nuclear transfer embryos. AB - The effects of activation by 6-dimethylaminopurine (6-DMAP) and cycloheximide (CHX) on the development and chromosomal complement of sheep parthenogenetic and SCNT embryos were investigated. The results revealed that the blastocyst development of parthenogenetic embryos was significantly higher (P < 0.05) in 6 DMAP activated oocytes, compared to those activated with CHX (21.0 +/- 0.9 vs. 14.9 +/- 0.5, respectively). In contrast, the blastocyst frequencies did not significantly differ (P > 0.05) between the two activation treatment groups for SCNT embryos. The 6-DMAP or CHX treatment did not result in any significant difference in the blastocyst total cell number in either parthenote or SCNT embryos. The chromosomal analysis revealed that all the parthenogenetic embryos (100.0%) derived from 6-DMAP treatment, were chromosomally abnormal whereas in CHX-treated embryos, it was significantly lowered (93.6%, P < 0.05). Conversely, the proportions of chromosomally abnormal SCNT embryos did not significantly differ (P > 0.05) among the 6-DMAP and CHX- treated embryo groups (60.0% vs. 56.2%, respectively). This study demonstrated that oocyte activation agents such as DMAP and CHX have differing effects on meiotic or mitotic nuclei. The study also highlighted the feasibility of using bovine X and Y chromosome specific painting probes in sheep embryos. PMID- 16211598 TI - Meiotic and epigenetic aberrations in Dnmt3L-deficient male germ cells. AB - The DNA methyltransferase-like protein Dnmt3L is necessary for the establishment of genomic imprints in oogenesis and for normal spermatogenesis (Bourc'his et al., 2001; Hata et al., 2002). Also, a paternally imprinted gene, H19, loses DNA methylation in Dnmt3L-/- spermatogonia (Bourc'his and Bestor, 2004; Kaneda et al., 2004). To determine the reason for the impaired spermatogenesis in the Dnmt3L-/- testes, we have carried out a series of histological and molecular studies. We show here that Dnmt3L-/- germ cells were arrested and died around the early meiotic stage. A microarray-based gene expression-profiling analysis revealed that various gonad-specific and/or sex-chromosome-linked genes were downregulated in the Dnmt3L-/- testes. In contrast, expression of retrovirus-like intracisternal A-particle (IAP) sequences was upregulated; consistent with this observation, a specific IAP copy showed complete loss of DNA methylation. These findings indicate that Dnmt3L regulates germ cell-specific gene expression and IAP suppression, which are critical for male germ cell proliferation and meiosis. PMID- 16211599 TI - Rat Spag5 associates in somatic cells with endoplasmic reticulum and microtubules but in spermatozoa with outer dense fibers. AB - The leucine zipper motif has been identified as an important and specific interaction motif used by various sperm tail proteins that localize to the outer dense fibers. We had found that rat Odf1, a major integral ODF protein, utilizes its leucine zipper to associate with Odf2, another major ODF protein, Spag4 which localizes to the interface between ODF and axonemal microtubule doublets, and Spag5. The rat Spag5 sequence indicated a close relationship with human Astrin, a microtubule-binding spindle protein suggesting that Spag5, like Spag4, may associate with the sperm tail axoneme. RT PCR assays indicated expression of Spag5 in various tissues and in somatic cells Spag5 localizes to endoplasmic reticulum and microtubules, as expected for an Astrin orthologue. MT binding was confirmed both in vivo and in in vitro MT-binding assays: somatic cells contain a 58 kDa MT-associated Spag5 protein. Western blotting assays of rat somatic cells and male germ cells at different stages of development using anti-Spag5 antibodies demonstrated that the protein expression pattern changes during spermatogenesis and that sperm tails contain a 58 kDa Spag5 protein. Use of affinity-purified anti-Spag5 antibodies in immuno electron microscopy shows that in rat elongated spermatids and epididymal sperm the Spag5 protein associates with ODF, but not with the axonemal MTs. This observation is in contrast to that for the other Odf1-binding, MT-binding protein Spag4, which is present between ODF and axoneme. Our data demonstrate that Spag5 has different localization in somatic versus male germ cells suggesting the possibility of different function. PMID- 16211600 TI - Suppression of translation during in vitro maturation of pig oocytes despite enhanced formation of cap-binding protein complex eIF4F and 4E-BP1 hyperphosphorylation. AB - In this study, we document that the overall rate of protein synthesis decreases during in vitro maturation (IVM) of pig oocytes despite enhanced formation of the 5' cap structure eIF4F. Within somatic/interphase cells, formation of the eIF4F protein complex correlates very well with overall rates of protein translation, and the formation of this complex is controlled primarily by the availability of the 5' cap binding protein eIF4E. We show that the eIF4E inhibitory protein, 4E BP1, becomes phosphorylated during IVM, which results in gradual release of eIF4E from 4E-BP1, as documented by immunoprecipitation analyses. Isoelectric focusing and Western blotting experiments show conclusively that eIF4E becomes gradually phosphorylated with a maximum at metaphase II (M II). The activity of eIF4E and its ability to bind mRNA also increases during oocyte maturation as documented in experiments with m7-methyl GTP-Sepharose, which mimics the cap structure of mRNA. Complementary analysis of flow-through fraction for 4E-BP1, and eIF4G proteins additionally provides evidence for enhanced formation of cap-binding protein complex eIF4F. Altogether, our results bring new insights to the regulation of translation initiation during meiotic division, and more specifically clarify that 4E-BP1 hyper-phosphorylation is not the cause of the observed suppression of overall translation rates. PMID- 16211601 TI - Eye-hand coordination in essential tremor. AB - Patients with essential tremor (ET) or with cerebellar lesions have in common oculomotor abnormalities, with the exception of saccadic eye movements, which do not seem to be involved in ET. Since grasping is prolonged in ET and might be related to saccadic dysmetria, we tested whether simultaneous hand pointing could unmask it. Twelve ET patients and 14 controls performed saccades with and without simultaneous pointing movements to the same targets, and with and without a gap between the disappearance of the fixation point and the appearance of the target. Eye movements were recorded with the magnetic search-coil method, hand movements with an ultrasound-emitting probe. ET patients did not have saccadic dysmetria, and contrary to normal subjects their saccadic latency did not decrease during combined eye-hand movements compared with saccades performed in isolation. Hand movements had a longer duration in ET patients, with decreased peak acceleration, an increased latency of the peak velocity, and peak deceleration. In conclusion, this first study on eye-hand coordination in ET revealed abnormal kinematic changes in the early phase of pointing movements. These changes might be related to cerebellar disease but they are independent of the intention tremor component and saccade performance. PMID- 16211602 TI - Effect of object transport on grasp coordination in multiple system atrophy. AB - We examined the effects of the parkinsonian variant of multiple-system atrophy (MSA-P) on grasp and forward transport and release of an object. Twelve patients with MSA-P and 10 age-matched control subjects performed the task with each of three object weights (200, 400, 800 gm). Subjects moved at a self-selected pace using a precision grip. The grip (normal) and load (tangential) forces and the object position were recorded. Results indicate subjects with MSA-P have temporal and force coordination deficits. Temporal delays were seen in all subjects with MSA-P, leading to prolonged overall movement times compared to control subjects. These delays occurred throughout the task, with significantly longer transport phases and delays releasing the object. Despite demonstrating an appropriate anticipatory scaling of forces, with increasing grip and load forces for heavier weights, force coordination was compromised in subjects with MSA-P. These subjects generated significant negative load forces prior to transporting the object. In addition, during the transport phase, subjects with MSA-P generated highly variable grip forces. Overall, the results indicate that subjects with MSA P demonstrate bradykinesia and difficulty coordinating components of an object transport task. PMID- 16211603 TI - Psychogenic palatal tremor. AB - We describe a case of psychogenic palatal tremor. The diagnosis was supported by clinical criteria and neurophysiological testing, including frequency analysis and jerk-locked back-averaging. We discuss the differential diagnosis of palatal tremor as well as the role of neurophysiological testing in the diagnosis of psychogenic movement disorders. PMID- 16211604 TI - Prevalence and clinical characteristics of restless legs syndrome in Japanese patients with Parkinson's disease. AB - To explore the clinical significance of restless legs syndrome (RLS) in Parkinson's disease (PD) and the causal relationship between these two disorders, we made a comparison of both the prevalence of RLS and the severity of sleep disturbance manifested on the Pittsburg Sleep Quality Index (PSQI) between patients with PD (n=165) and age- and sex-matched control subjects (n=131). The prevalence of RLS diagnosed by clinical interview was significantly higher in PD patients than in control subjects (12% vs. 2.3%). PSQI score was significantly higher in PD patients with RLS than in both patients without RLS and controls. However, PSQI score was not statistically different between the latter two groups. Among the PD patients with RLS, only 2 had a positive family history of RLS. Only 3 PD patients had requested treatment for the disorder. Our results emphasize the etiological link between RLS and PD in a Japanese cohort, and the existence of RLS is thought to be one of the most important factors aggravating sleep disturbance in PD, despite the low RLS severity. PMID- 16211605 TI - Functional correlates of mild parkinsonian signs in the community-dwelling elderly: poor balance and inability to ambulate independently. AB - Mild tremor, rigidity, and bradykinesia (mild parkinsonian signs, MPS) are commonly detected during the clinical examination of elderly people without known neurological disease. The functional correlates of these incidental findings are not well understood. Balance and ability to ambulate independently are important functions in the elderly. The objective of this study is to examine whether MPS were associated with impaired balance. Balance was assessed using a subjective measure (complaint of poor balance) and a functional measure (the need to use a walker, cane, or wheelchair). Our methods included the neurological evaluation of nondemented older people in Washington Heights-Inwood, NY. Of 2,251 participants, 527 (23.4%) complained of poor balance; 538 (23.9%) required a cane, walker, or wheelchair; and 363 (16.1%) had MPS. In adjusted logistic regression analyses, MPS were associated with a complaint of poor balance (OR=1.5, 95% CI=1.1-2.0) and the need to use a cane, walker, or wheelchair (OR=1.9, 95% CI=1.4-2.6). The need to use a cane, walker, or wheelchair was associated with changes in axial function (OR=5.5, 95% CI=2.6-11.6) as well as rigidity (OR=1.5, 95% CI=1.07-2.2). Although they are incidental and subtle, signs of bradykinesia, rigidity, and tremor are associated with impaired function. The elderly in whom these signs have been detected are 50% more likely to complain of poor balance and 90% more likely to require a cane, walker, or wheelchair than are their counterparts without these signs. PMID- 16211606 TI - Unusual position-sensitive jaw tremor responsive to botulinum toxin. AB - We report on a woman with a focal position-sensitive jaw tremor present with the jaw held slightly open or while drinking from a cup. Electromyography demonstrated a rhythmic 5 Hz alternating tremor involving masseter and digastric muscles with normal reciprocal inhibition. The tremor was markedly reduced with botulinum toxin injections. PMID- 16211607 TI - Applications of X-ray powder diffraction in materials chemistry. AB - X-ray powder diffraction is a standard technique in materials chemistry, yet it is often still used in the laboratory as a "one-hit" technique, e.g. for fingerprinting and following the progress of reactions. It is important, however, that the wealth of information available from powder data is not overlooked. While it is only possible here to scratch the surface of possibilities, a range of examples from our research is used to emphasize some of the more accessible techniques and to highlight successes as well as potential problems. The first example is the study of solid solution formation in the oxide systems Ba(3 3x)La(2x)V2O8 and Sr(4-x)Ba(x)Mn3O10 and in the silicate-hydroxyapatite bioceramic, Ca10(PO4)6-x(SiO4)x(OH)2-x. Database mining is also explored, using three phases within the pseudobinary phase diagram Li3SbO4-CuO as examples. All three phases presented different challenges: the structure of Li3SbO4 had been previously reported in higher symmetry than was actually the case, Li3Cu2SbO6 was found to be isostructural with Li2TiO3 but the cation ordering had to be rationalized, and Li3CuSbO5 was believed to be triclinic, presenting challenges in indexing the powder pattern. Quantitative phase analysis is briefly discussed, with the emphasis both on success (determination of amorphous phase content in a novel cadmium arsenate phase) and on possible failure (compositional analysis in bone mineral); the reasons for the problems in the latter are also explored. Finally, the use of an area detector system has been shown to be of value in the study of orientational effects (or lack of them) in non- and partially-ordered biomaterials, including p-HEMA, annulus fibrosis of lumbar discs, and keratin in the horn of cow's hooves. PMID- 16211608 TI - Huntington's disease patients have selective problems with insight. AB - The objective of this study was to determine insight in patients with Huntington's disease (HD) by contrasting patients' ability to rate their own behavior with their ability to rate a person other than themselves. HD patients and carers completed the Dysexecutive Questionnaire (DEX), rating themselves and each other at two time points. The temporal stability of these ratings was initially examined using these two time points since there is no published test retest reliability of the DEX with this population to date. This was followed by a comparison of patients' self-ratings and carer's independent ratings of patients by performing correlations with patients' disease variables, and an exploratory factor analysis was conducted on both sets of ratings. The DEX showed good test-retest reliability, with patients consistently and persistently underestimating the degree of their dysexecutive behavior, but not that of their carers. Patients' self-ratings and carers' ratings of patients both showed that dysexecutive behavior in HD can be fractionated into three underlying components (Cognition, Self-regulation, Insight), and the relative ranking of these factors was similar for both data sets. HD patients consistently underestimated the extent of only their own dysexecutive behaviors relative to carers' ratings by 26%, but were similar in ascribing ranks to the components of dysexecutive behavior. PMID- 16211610 TI - Prevalence study of primary dystonia in Iceland. AB - In Iceland, the crude prevalence for all types of primary dystonia was 37.1/10(5) (confidence interval, 30.4-44.9). Focal dystonia had the highest prevalence (31.2/10(5)), followed by segmental (3.1/10(5)), multifocal (2.4/10(5)) and generalized dystonia (0.3/10(5)). Cervical dystonia was the most common focal dystonia (11.5/10(5)), followed by limb dystonia (8.0/10(5)), laryngeal dystonia (5.9/10(5)), blepharospasm (3.1/10(5)), and oromandibular dystonia (2.8/10(5)). The male:female ratio for all patients was 1:1.9 (P=0.0007), and females outnumbered males in all subtypes except oromandibular dystonia. Mean age of onset for all patients was 42.7 years (range, 3-82 years). This prevalence of primary dystonia is higher than in most reported studies, possibly because of more complete ascertainment but the relative frequencies of dystonia subtypes is similar. PMID- 16211609 TI - p.H1069Q mutation in ATP7B and biochemical parameters of copper metabolism and clinical manifestation of Wilson's disease. AB - We compared the effect of the p.H1069Q mutation and other non-p.H1069Q mutations in ATP7B on the phenotypic expression of Wilson's disease (WD), and assessed whether the clinical phenotype of WD in compound heterozygotes depends on the type of mutation coexisting with the p.H1069Q. One hundred forty-two patients with clinically, biochemically, and genetically diagnosed WD were studied. The mutational analysis of ATP7B was performed by direct sequencing. A total number of 26 mutations in ATP7B were identified. The p.His1069Gln was the most common mutation (allelic frequency: 72%). Seventy-three patients were homozygous for this mutation. Of compound heterozygotes, 37 had frameshift/nonsense mutation, and 20 had other missense mutation on one of their ATP7B alleles. Twelve patients had two non-p.H1069Q mutations. Patients homozygous for the p.H1069Q mutation had the less severe disturbances of copper metabolism and the latest presentation of first WD symptoms. The most severely disturbed copper metabolism and the earliest age at initial disease manifestation was noticed in non-p.H1069Q patients. In compound heterozygotes, the type of mutation coexisting with the p.H1069Q to a small extent influenced WD phenotype. The phenotype of WD varied considerably among patients with the same genotype. The p.H1069Q mutation is associated with late WD manifestation and with a mild disruption of copper metabolism. In compound heterozygotes, the phenotype of WD to a small extent depends on the type of mutation coexisting with the p.H1069Q. Besides genotype, additional modifying factors seem to determine WD manifestations. PMID- 16211611 TI - Camptocormia associated with focal myositis in multiple-system atrophy. AB - Camptocormia (CC) or pronounced forward flexion of the trunk is a common symptom of Parkinson's disease. We describe 2 patients with probable, respectively possible multiple-system atrophy and CC. Magnetic resonance imaging of the erector trunci showed focal patchy hyperintensities with gadolinium enhancement and muscle biopsy was indicative of variably pronounced focal myositis. CC was progressive and the major handicap for both patients after 1 and 1.5 years of follow-up, respectively. The therapeutic response was poor. Similarities with the dropped-head syndrome suggest that the muscle pathology may be either the primary cause of CC, a focal reaction to the CC posture, or a coincident syndrome of old age. PMID- 16211612 TI - Positron emission tomography changes in PARK1 mutation. PMID- 16211613 TI - HS1-BP3 gene variant is common in familial essential tremor. AB - Essential tremor (ET) is a movement disorder characterized by a postural or kinetic tremor of the hands, head, or voice. It is typically a familial condition and affects 1% to 4% of the general population. The trait is genetically linked to chromosome 2p in some families. A variant (828C-->G) in exon 7 of the hematopoietic-specific protein 1 binding protein 3 gene (HS1-BP3) on chromosome 2p recently has been found to segregate with ET in 2 families. To determine the frequency of this variant in a larger series, we studied patients with ET, Parkinson disease (PD), and controls without tremor. Affected singletons representing 73 families from the United States with dominantly inherited ET, 35 individuals with PD, and 304 healthy controls older than age 60 were tested for the 828C-->G variant in exon 7 of the HS1-BP3 gene by a BseYI restriction enzyme digest of the polymerase chain reaction product. Heterozygous carriers of the mutant allele were identified in 12 individuals with ET (16.4%) and in 1 individual with PD and postural tremor (3%). All of the healthy controls (608 chromosomes) were homozygous for the wild-type allele. The 828C-->G genetic variant in the HS1-BP3 gene occurs relatively frequently in subjects with ET. The variant may also be found in some individuals with PD and postural tremor. The HS1-BP3 gene plays a putative role in regulating catecholamine and serotonin metabolism, but the functional consequences of the amino acid substitution (A265G) caused by this genetic variant is unknown. PMID- 16211614 TI - Belly dance syndrome due to spinal myoclonus. AB - We report on a case of spinal myoclonus resembling a belly dance syndrome. PMID- 16211615 TI - Spinocerebellar ataxia associated with a mutation in the fibroblast growth factor 14 gene (SCA27): A new phenotype. AB - Autosomal dominant cerebellar ataxias (ADCAs) are genetically classified into spinocerebellar ataxias (SCAs). We describe 14 patients of a Dutch pedigree displaying a distinct SCA-phenotype (SCA27) associated with a F145S mutation in the fibroblast growth factor 14 (FGF14) gene on chromosome 13q34. The patients showed a childhood-onset postural tremor and a slowly progressive ataxia evolving from young adulthood. Dyskinesia was often present, suggesting basal ganglia involvement, which was supported by functional imaging in 1 patient. Magnetic resonance imaging (MRI) of the brain showed only moderate cerebellar atrophy in the 2 eldest patients. Neuropsychological testing indicated low IQ and deficits in memory and executive functioning. Behavioral problems were also observed. Further investigations will have to determine the role of FGF14 in the pathogenesis of neurodegeneration and the frequency of this FGF14 mutation in SCA. (c) 2005 Movement Disorder Society. PMID- 16211616 TI - Learning a high-precision locomotor task in patients with Parkinson's disease. AB - We evaluated the acquisition and performance of a high-precision locomotor task in patients with Parkinson's disease (PD) and healthy subjects. All subjects walked on a treadmill and had to step repetitively as low as possible over an obstacle without touching it. During blocks 1 and 2, the subjects had full vision and received additional acoustic warning and feedback signals. During block 3, vision became restricted. Changes in foot clearance and the number of obstacle hits were evaluated. Initially, PD patients performed poorer and improved foot clearance slower. After task repetition, the groups performed similarly. Restricting vision deteriorated performance in both groups. The similar performance of PD patients after task repetition might indicate that adequate training could improve adaptive locomotor behavior in PD patients. PMID- 16211617 TI - Uniform qualitative electrophysiological changes in postoperative rest tremor. AB - Ablation and deep brain stimulation (DBS) can treat pharmacologically uncontrollable tremor. Here, we compared the postoperative electrophysiological changes in resting hand tremor after 32 ablations and 12 DBS implantations in patients with severe tremor-dominant idiopathic Parkinson's disease (PD) and essential tremor (ET). Short- and long-term accelerometric data were acquired after surgery and were compared to the preoperative tremor. After effective surgical treatments, significant rest tremor reduction and increase in both frequency and approximate entropy (ApEn) were detected in all PD cases, irrespective of the type and target of intervention. However, the long-term effect of DBS implantation on tremor reduction was significantly better compared to that after ablative treatments. In cases of thalamotomy, the postoperative increase in frequency and ApEn was significantly larger in essential tremor compared to PD, suggesting that the etiology of tremor may influence the size of the similar changes. However, cases where clinical tremor re-emerged 6 to 12 months after the surgery, no change in frequency and ApEn was detected on the second postoperative day, despite an initial tremor reduction and clinical improvement similar to the effective operations. Our results suggest that uniform postoperative changes in rest tremor and the increase in frequency and ApEn could be due to attenuation of pathological oscillators and might be immediate indicators of the effectiveness of neurosurgical treatments relieving tremor. PMID- 16211619 TI - On-demand deep brain stimulation for essential tremor: a report on four cases. AB - Deep brain stimulation (DBS) is an established therapy for essential tremor (ET), but loss of efficacy due to tolerance can occur. Our objective was to evaluate if it is feasible to use DBS only on-demand and if this would prevent tolerance. We report on the effects of left-side thalamic DBS in 4 ET patients who were instructed to switch on stimulation only when using their right hand for motor tasks and were followed-up to 30 months after surgery. The patients were capable of using DBS only on-demand (DBS use of 22.0+/-13.5%/day). DBS led to a stable suppression of right arm tremor throughout the follow-up. No problems associated with tolerance such as tremor rebound or late therapy failure occurred. In comparison to publications stating that ET patients had been using DBS continuously during the daytime, the use of on-demand DBS saves battery life, which delays surgical replacement of the stimulator. Thus, on-demand DBS saves money, may help to prevent tolerance, and should be adopted for the long-term treatment of ET patients. PMID- 16211618 TI - Placebo-controlled study of rTMS for the treatment of Parkinson's disease. AB - The objective of this study is to assess the safety and efficacy of repetitive transcranial magnetic stimulation (rTMS) for gait and bradykinesia in patients with Parkinson's disease (PD). In a double-blind placebo-controlled study, we evaluated the effects of 25 Hz rTMS in 18 PD patients. Eight rTMS sessions were performed over a 4-week period. Four cortical targets (left and right motor and dorsolateral prefrontal cortex) were stimulated in each session, with 300 pulses each, 100% of motor threshold intensity. Left motor cortex (MC) excitability was assessed using motor evoked potentials (MEPs) from the abductor pollicis brevis. During the 4 weeks, times for executing walking and complex hand movements tests gradually decreased. The therapeutic rTMS effect lasted for at least 1 month after treatment ended. Right-hand bradykinesia improvement correlated with increased MEP amplitude evoked by left MC rTMS after individual sessions, but improvement overall did not correlate with MC excitability. rTMS sessions appear to have a cumulative benefit for improving gait, as well as reducing upper limb bradykinesia in PD patients. Although short-term benefit may be due to MC excitability enhancement, the mechanism of cumulative benefit must have another explanation. PMID- 16211620 TI - A blessing in disguise: resolution of tardive dyskinesia with development of cervical myelitis. AB - Tardive dyskinesia (TD), which is frequently seen in patients treated with dopamine receptor blocking agents, is difficult to manage. We report on a young Chinese man with bipolar disorder who developed TD after haloperidol treatment, involving the trunk, limbs, and orofacial area. TD persisted despite switching to atypical antipsychotic agents and treatment with valproate, benzodiazepines, and tetrabenazine. Resolution only occurred years later when he developed quadriplegia arising from infective myelitis of the cervical cord (C4-5). He had concomitant vertebral osteomyelitis, which was successfully treated with intravenous antibiotics. With intensive rehabilitation, he recovered the use of his limbs, but had no recurrence of TD. We attribute the resolution of orofacial dyskinesias with a cervical lesion to the interconnections between the orofacial area and cervical spine via the trigeminal nucleus (which has fibers descending as far caudally as C6), as well as to resetting of cortical maps. PMID- 16211621 TI - Economic burden associated with Parkinson's disease on elderly Medicare beneficiaries. AB - We evaluated medical utilization and economic burden of self-reported Parkinson's disease (PD) on patients and society. Using the 1992-2000 Medicare Current Beneficiary Survey, we compared health care utilization and expenditures (in 2002 U.S. dollars) of Medicare subscribers with and without PD, adjusting for sociodemographic characteristics and comorbidities. PD patients used significantly more health care services of all categories and paid significantly more out of pocket for their medical services than other elderly (mean+/- SE, 5,532 dollars+/- 329 dollars vs. 2,187 dollars+/- 38 dollars; P<0.001). After adjusting for other factors, PD patients had higher annual health care expenses than beneficiaries without PD (18,528 dollars vs. 10,818 dollars; P<0.001). PD patients were more likely to use medical care (OR=3.77; 95% CI=1.44-9.88), in particular for long-term care (OR=3.80; 95% CI=3.02-4.79) and home health care (OR=2.08; 95% CI=1.76-2.46). PD is associated with a significant economic burden to patients and society. Although more research is needed to understand the relationship between PD and medical expenditures and utilization, these findings have important implications for health care providers and payers that serve PD populations. PMID- 16211622 TI - Power sources for portable electronics and hybrid cars: lithium batteries and fuel cells. AB - The activities in progress in our laboratory for the development of batteries and fuel cells for portable electronics and hybrid car applications are reviewed and discussed. In the case of lithium batteries, the research has been mainly focused on the characterization of new electrode and electrolyte materials. Results related to disordered carbon anodes and improved, solvent-free, as well as gel type, polymer electrolytes are particularly stressed. It is shown that the use of proper gel electrolytes, in combination with suitable electrode couples, allows the development of new types of safe, reliable, and low-cost lithium ion batteries which appear to be very promising power sources for hybrid vehicles. Some of the technologies proven to be successful in the lithium battery area are readapted for use in fuel cells. In particular, this approach has been followed for the preparation of low-cost and stable protonic membranes to be proposed as an alternative to the expensive, perfluorosulfonic membranes presently used in polymer electrolyte membrane fuel cells (PEMFCs). PMID- 16211624 TI - Evolutionary relationship and application of a superfamily of cyclic amidohydrolase enzymes. AB - Cyclic amidohydrolases belong to a superfamily of enzymes that catalyze the hydrolysis of cyclic C-N bonds. They are commonly found in nucleotide metabolism of purine and pyrimidine. These enzymes share similar catalytic mechanisms and show considerable structural homologies, suggesting that they might have evolved from a common ancestral protein. Homology searches based on common mechanistic properties and three-dimensional protein structures provide clues to the evolutionary relationships of these enzymes. Among the superfamily of enzymes, hydantoinase has been highlighted by its potential for biotechnological applications in the production of unnatural amino acids. The enzymatic process for the production of optically pure amino acids consists of three enzyme steps: hydantoin racemase, hydantoinase, and N-carbamoylase. For efficient industrial application, some critical catalytic properties such as thermostability, catalytic activity, enantioselectivity, and substrate specificity require further improvement. To this end, isolation of new enzymes with desirable properties from natural sources and the optimization of enzymatic processes were attempted. A combination of directed evolution techniques and rational design approaches has made brilliant progress in the redesign of industrially important catalytic enzymes; this approach is likely to be widely applied to the creation of designer enzymes with desirable catalytic properties. PMID- 16211623 TI - Development of protein, peptide, and small molecule catalysts using catalysis based selection strategies. AB - We have developed peptide catalysts and antibody catalysts that catalyze aldol, retro-aldol, and Michael reactions via an enamine mechanism using reaction-based selections with 1,3-diketone derivatives. Nucleophilic amino groups of the catalysts were covalently trapped during the selections. We have also developed fluorogenic substrates that are useful for real-time monitoring of the progress of bond-forming reactions, such as aldol reactions, by an increase in fluorescence. These fluorogenic substrates have been used to monitor peptide catalyzed, antibody-catalyzed, enzyme-catalyzed, and small molecule-catalyzed reactions. Catalysis-based screening using fluorogenic substrates will accelerate rapid identification of superior catalysts and reaction conditions. PMID- 16211625 TI - Family physician consultation patterns indicate high risk for early-onset anorexia nervosa. AB - OBJECTIVE: There is often a delay in the recognition of early-onset anorexia nervosa. The current study aimed to determine whether there are specific patterns in the frequency and content of family physician consultations that might predict its onset. METHOD: Lifetime number and type of family physician consultations were recorded for three groups: (a) an index group comprising 19 girls with anorexia nervosa, onset under 14; (b) a clinical control group comprising 19 girls with an emotional disorder; and (c) a nonclinical group comprising 19 girls with no history of mental health problems. RESULTS: Both clinical groups had an elevated number of consultations, particularly in the 5 years before diagnosis. The index group had a significantly higher number of eating, weight, and shape consultations (especially in the year before diagnosis), whereas the clinical control group had a greater number of psychological consultations. CONCLUSION: A single consultation about eating behaviour or weight and shape concerns is a strong predictor of the subsequent emergence of anorexia nervosa. PMID- 16211626 TI - Impulsivity and compulsivity in bulimia nervosa. AB - OBJECTIVE: A primary objective of the current article is to investigate the relationship between impulsivity and compulsivity in patients with bulimia nervosa (BN). A second goal is to explore the relationship between impulsivity and compulsivity and related psychiatric problems. METHOD: Two-hundred four females with BN completed several measures of impulsivity and compulsivity as well as measures of personality, substance use, eating pathology, and depression. RESULTS: Participants reported considerable variability on measures of impulsivity and compulsivity and these scores were positively correlated with each other. Impulsive-compulsive groups differed in personality, substance use, eating, and depression. CONCLUSION: These findings suggest that impulsivity and compulsivity can coexist in BN patients and that both traits may provide useful information about comorbid problems in women with BN. PMID- 16211627 TI - Problematic eating and feeding behaviors of 36-month-old children. AB - OBJECTIVE: We incorporated selected items from several existing instruments to create a comprehensive multifactorial instrument to measure problematic eating behaviors in young children and to examine the prevalences and correlates of these behaviors. METHOD: A community sample of young mothers (N = 93) completed the inventory of problematic eating behaviors for their 36-month-old children. RESULTS: The most common child problems reported by mothers were the spitting out of food during feedings and becoming upset when food was restricted. A four factor solution identified pickiness (e.g., child eats a limited variety of food), food refusal (e.g., child refuses to eat specific foods), struggle for control (e.g., frequent struggles with child over food), and positive parental behavior (e.g., praising child about his/her food intake) domains. Internal consistency was moderate to good for all factors. Only the struggle for control factor was related to other problematic behaviors as measured by the Child Behavior Check List (CBCL). The food refusal factor was related to mothers' lifetime history of any psychopathology. The pickiness factor was related to mothers' lifetime history of alcohol dependence. CONCLUSION: Findings suggest that child clinicians should be sensitive to the quality of mother-child interactions during feeding. PMID- 16211628 TI - Influence of race, gender, and socioeconomic status on binge eating frequency in a population-based sample. AB - OBJECTIVE: This study investigates race and gender differences in the demographic and socioeconomic determinants of frequency of binge eating using a population based sample. METHOD: An analysis of self-reported data on 573 women and 360 men (range, 18-97 years) from a cross-sectional, multistage area probability sample of individuals aged 18 years and older residing in the Detroit metropolitan area conducted in 1995. RESULTS: For women, the frequency of binge eating is negatively associated with age and family income, and positively associated with being married, depression, and time residing in polluted neighborhoods. For men, the frequency of binge eating is negatively associated with age. The frequency of binge eating was not affected by education, race, obesity, or current dieting. CONCLUSION: The frequency of binge eating is highest among adults younger than 40 years. It follows an income gradient for women, but not for men. After controls for individual disadvantage, there is no residual Black/White difference in binge eating. PMID- 16211629 TI - Purging disorder: an ominous variant of bulimia nervosa? AB - OBJECTIVE: The current study evaluates the clinical significance and distinctiveness of purging disorder (PD), an eating disorder characterized by recurrent purging in the absence of objective binge episodes (OBE) among normal weight individuals. METHOD: Women with PD (n = 37), bulimia nervosa (BN; n = 39), or no eating disorder (n = 35) completed clinical assessments. Women with PD (n = 23) and BN (n = 25) completed 6-month follow-up assessments. RESULTS: Compared with controls, both eating-disordered groups reported significantly higher eating, Axis I, and Axis II pathology. Compared with BN, PD was associated with significantly lower eating concerns, disinhibition, and hunger. At 6-month follow up, rates of remission did not differ significantly between PD and BN, and crossover between disorders was rare. CONCLUSION: PD appears to be a clinically significant and potentially distinctive eating disorder. PMID- 16211630 TI - Parents' reports of the body shape and feeding habits of 36-month-old children: an investigation of gender differences. AB - OBJECTIVE: The current study examined parental perception of offspring body shape, differential reporting of offspring eating behaviors by mothers and fathers, and gender-specific patterns of offspring feeding habits. METHODS: Parents of a community sample of 36-month-old children (N = 93) completed measures regarding their offspring's feeding patterns and body shape. RESULTS: Results revealed noteworthy correlates (e.g., concerns about their child's appetite) of parental perception of offspring weight status. They further suggested that mothers and fathers often differed in their accounts of their child's eating habits, and that parents report certain eating behaviors differently depending on the gender of their child. CONCLUSION: Clinical and theoretical implications are discussed. PMID- 16211631 TI - Ethnicity and sampling bias in binge eating disorder: Black women who seek treatment have different characteristics than those who do not. AB - OBJECTIVE: The current study investigated sampling bias as it affects recruited clinic samples of Black and White women with binge eating disorder (BED). METHODS: Clinical characteristics of a recruited clinic sample (35 Black and 302 White consecutively evaluated women) with BED were compared with a community sample of Black and White women with BED drawn from the New England Women's Health Project. The clinic and community groups met the same definition of BED and were assessed with identical methods. RESULTS: Among White women, the clinic and community samples differed on some features (higher body mass index [BMI], and greater eating concerns and shape concerns in the clinic sample) but these differences reflected small to moderate effects sizes. In contrast, among Black women, the clinic sample had substantially higher levels (large effect sizes) of several features of eating disorders (eating concerns, dietary restraint, and shape concern), higher (moderate effect size) BMI, but lower frequency of binge eating (moderate effect size) than the community sample. A comparison of Black and White women within the clinic sample revealed little difference in clinical presentation, except for the significantly higher BMI among Black women. CONCLUSION: A sampling bias appears to exist between both White and Black recruited clinic samples of women with BED, although the bias appears to be substantially greater for Black women. Black women with BED who seek treatment have significantly different characteristics than those who do not. PMID- 16211632 TI - Disordered eating attitudes and behaviors in ballet students: examination of environmental and individual risk factors. AB - OBJECTIVE: The current study compared the prevalence of disordered eating attitudes and behaviors among adolescent ballet dancers at national, regional, and local schools. METHOD: Female ballet students (N = 239; mean age = 15.0 +/- 1.5 years) from five geographically disparate summer programs completed the Eating Dis-order Inventory (EDI) and answered questions regarding eating disorder symptoms. RESULTS: Students from both national and local schools reported significantly higher EDI total, Drive for Thinness, and Perfectionism scores compared with regional students. In contrast, national students reported significantly greater dieting scores and lifetime histories of self-induced vomiting compared with regional and local students. CONCLUSION: Eating pathology among adolescent ballet dancers may be a function of both genetic and environmental risk. Dancers who exhibit high levels of perfectionism and, perhaps consequently, place themselves in highly competitive environments, may exhibit a significantly increased risk for disordered eating in comparison to dancers who are less perfectionistic and/or place themselves in less competitive environments. PMID- 16211633 TI - Can we simultaneously work toward the prevention of obesity and eating disorders in children and adolescents? AB - Because of the growing prevalence of obesity among children and adolescents, increased attention is being directed toward its prevention. An important question is: can we simultaneously work toward the prevention of obesity and eating disorders? To address this question, we need to determine (a) if there is a need for integrated approaches; (b) if we can bridge the fields of obesity and eating disorders; (c) if we can foster the development of environments that promote healthy eating and physical activity choices and the acceptance of diverse body shapes and sizes; and (d) if we can work toward the development of interventions that have relevance to a broad spectrum of weight-related conditions and behaviors. PMID- 16211634 TI - Advanced anorexia nervosa, associated with pneumomediastinum, pneumothorax, and soft-tissue emphysema without esophageal lesion. AB - OBJECTIVE: We describe the case of a 20-year-old female patient suffering from anorexia nervosa since the age 15 of years. When admitted, she was 168 cm tall with a weight of 35 kg (body mass index [BMI] = 12.4). A week after admission, the patient complained of retrosternal pain, and a few hours later she developed extensive soft-tissue emphysema. METHODS: The X-ray picture of her thorax showed left-hand supraclavicular and infraclavicular emphysema, extensive emphysema in both sides of the soft tissue of the throat, a left-hand apical pneumothorax (4 mm), and a pneumomediastinum. RESULTS: We discuss the extent to which these symptoms are interrelated in the sense of a causal nexus. CONCLUSION: We ruled out any other differential-diagnostic etiopathogenic factors and concluded that the cause of our patient's soft-tissue emphysema, pneumothorax, and pneumomediastinum might well be autodestructive behavior. PMID- 16211635 TI - Seasonally bound ovopathy versus "temperature at conception" as cause for anorexia nervosa and other eating disorders. AB - OBJECTIVE: A preponderance of births between April and June in patients with anorexia nervosa (AN) and other eating disorders (EDs) has recently been explained by a higher environmental temperature at conception. This hypothesis, however, does not explain some other irregularities in the month of birth distribution in contrast to the seasonal preovulatory overripeness ovopathy (SPrOO) and seasonal optimally ripened oocytes (SOptRO) hypotheses. METHOD: The month of birth distribution of 408 patients with EDs from Southeast England and of 199 patients from Australia has been reassessed in relation to previously determined high-risk and low-risk months based on a priori knowledge from mammals and on the physiologic birth peaks and troughs in humans at population level. The differences between high-risk and low-risk months were assessed by conventional chi-square analyses and by relative risks (RRs) with 95% confidence intervals (CI). RESULTS: In England, fewer AN patients (n = 259) were born at the peaks of the total birth pattern, that is, during the 2 low-risk months, and more were born during the transitional stages, that is, during the remaining 10 high-risk months (p = .004; RR = 1.66 [95% CI = 1.13-2.41]). After the addition of 149 other ED patients (n = 408), this result remained (p = .005; RR = 1.47 [95% CI = 1.10-1.97]). Similar results were found in 199 ED patients from Australia (p = .009; RR =1.70 [95% CI = 1.09-2.64]). CONCLUSION: ED patients appear to be more frequently conceived during the transitional stages of the ovulatory pattern due to SPrOO and less frequently at the ovulatory peaks due to SOptRO. The pathophysiologic processes of oocyte maturation in estrous animals and the circumstantial evidence of similar phenomena in humans are more appropriate to explain these coincidences than the temperature at conception hypothesis. PMID- 16211636 TI - Morphomolecular neuronal phenotypes in the neocortex reflect phylogenetic relationships among certain mammalian orders. AB - The cytoarchitecture of the cerebral cortex in mammals has been traditionally investigated using Nissl, Golgi, or myelin stains and there are few comparative studies on the relationships between neuronal morphology and neurochemical specialization. Most available studies on neuronal subtypes identified by their molecular and morphologic characteristics have been performed in species commonly used in laboratory research such as the rat, mouse, cat, and macaque monkey, as well as in autopsic human brain specimens. A number of cellular markers, such as neurotransmitters, structural proteins, and calcium-buffering proteins, display a highly specific distribution in distinct classes of neocortical neurons in a large number of mammalian species. In this article, we present an overview of the morphologic characteristics and distribution of three calcium-binding proteins, parvalbumin, calbindin, and calretinin, and of a component of the neuronal cytoskeleton, nonphosphorylated neurofilament protein in the neocortex of various species, representative of the major subdivisions of mammals. The distribution of these neurochemical markers defined several species- and order-specific patterns that permit assessment of the degree to which neuronal morphomolecular specialization, as well as the regional and laminar distribution of distinct cell types in the neocortex, represents derived or ancestral features. In spite of the remarkable diversity in morphologic and cellular organization that occurred during mammalian neocortical evolution, such patterns identified several associations among taxa that closely match their phylogenetic relationships. PMID- 16211638 TI - A case of uterine hematoma presenting as a transient vascular abnormality on ultrasonography. PMID- 16211637 TI - Anatomical analysis of an aye-aye brain (Daubentonia madagascariensis, primates: Prosimii) combining histology, structural magnetic resonance imaging, and diffusion-tensor imaging. AB - This report presents initial results of a multimodal analysis of tissue volume and microstructure in the brain of an aye-aye (Daubentonia madagascariensis). The left hemisphere of an aye-aye brain was scanned using T2-weighted structural magnetic resonance imaging (MRI) and diffusion-tensor imaging (DTI) prior to histological processing and staining for Nissl substance and myelinated fibers. The objectives of the experiment were to estimate the volume of gross brain regions for comparison with published data on other prosimians and to validate DTI data on fiber anisotropy with histological measurements of fiber spread. Measurements of brain structure volumes in the specimen are consistent with those reported in the literature: the aye-aye has a very large brain for its body size, a reduced volume of visual structures (V1 and LGN), and an increased volume of the olfactory lobe. This trade-off between visual and olfactory reliance is likely a reflection of the nocturnal extractive foraging behavior practiced by Daubentonia. Additionally, frontal cortex volume is large in the aye-aye, a feature that may also be related to its complex foraging behavior and sensorimotor demands. Analysis of DTI data in the anterior cingulum bundle demonstrates a strong correlation between fiber spread as measured from histological sections and fiber spread as measured from DTI. These results represent the first quantitative comparison of DTI data and fiber-stained histology in the brain. PMID- 16211639 TI - Proline-based macrocyclic inhibitors of the hepatitis C virus: stereoselective synthesis and biological activity. PMID- 16211640 TI - Sandwich immunoassay as a high-throughput screening method for cross-coupling reactions. PMID- 16211641 TI - Nickel-foam-supported reticular CoO-Li2O composite anode materials for lithium ion batteries. PMID- 16211642 TI - Hysteretic magnetic bistability based on a molecular azide switch. PMID- 16211643 TI - Catalysis by gold: isolated surface Au3+ ions are active sites for selective hydrogenation of 1,3-butadiene over Au/ZrO2 catalysts. PMID- 16211644 TI - Hydrogen-bond-mediated self-assembly of aminopyrazolones: macrocyclic quartets single and stacked one-dimensional motifs. PMID- 16211645 TI - Mild tricuspid regurgitation: a benign fetal finding at various stages of pregnancy. AB - OBJECTIVE: Tricuspid regurgitation (TR) may accompany various anatomical malformations and/or dysfunction of the fetal right heart. It may also appear in an anatomically healthy heart. With improved ultrasound modalities, more cases than the previously estimated prevalence of fetal TR in the low-risk population are being diagnosed. The objective of this study was to determine the prevalence of mild fetal TR in a low-risk obstetric population. METHODS: In 157 low-risk pregnant women (age range, 18-42 years) undergoing both early second-trimester and mid-trimester targeted organ scanning, including complete fetal echocardiography according to the five transverse planes technique, the apical four-chamber view was visualized using gray-scale, color Doppler and spatiotemporal image correlation (STIC) ultrasound modalities, with optimal acquisition parameters. RESULTS: Mild-to-moderate TR was discovered in the early second-trimester scan in 131/157 (83.4%) fetuses. No cases of cardiac malformation were found. All fetuses showed normal flow in the ductus venosus, including in one case diagnosed with moderate TR. Only in 39 (24.8%) cases was mild TR still evident at the second, mid-trimester scan. Neonatal echocardiography revealed mild TR in eight (5.1%) cases. No cases of chromosomal anomalies were detected. CONCLUSION: Mild TR is a benign finding of a temporal nature in early pregnancy. PMID- 16211646 TI - Congenital pulmonary airway malformation and congenital varicella infection-a possible association. PMID- 16211647 TI - Three-dimensional tissue block ultrasound in ovarian tumors. PMID- 16211650 TI - Single-site heterogeneous catalysts. AB - Intellectually, the advantages that flow from the availability of single-site heterogeneous catalysts (SSHC) are many. They facilitate the determination of the kinetics and mechanism of catalytic turnover-both experimentally and computationally-and make accessible the energetics of various intermediates (including short-lived transition states). These facts in turn offer a rational strategic principle for the design of new catalysts and the improvement of existing ones. It is generally possible to prepare soluble molecular fragments that circumscribe the single-site, thus enabling a direct comparison to be made, experimentally, between the catalytic performance of the same active site when functioning as a heterogeneous (continuous solid) as well as a homogeneous (dispersed molecular) catalyst. This approach also makes it possible to modify the immediate atomic environment as well as the central atomic structure of the active site. From the practical standpoint, SSHC exhibit very high selectivities leading to the production of sharply defined molecular products, just as do their homogeneous analogues. Given that mesoporous silicas with very large internal surface areas are ideal supports for SSHC, and that more than a quarter of the elements of the Periodic Table may be grafted as active sites onto such silicas, there is abundant scope for creating new catalytic opportunities. PMID- 16211651 TI - Two-step spin conversion in a cyanide-bridged ferrous square. PMID- 16211654 TI - Non-nucleoside HIV reverse transcriptase inhibitors, Part 6[1]: synthesis and anti-HIV activity of novel 2-[(arylcarbonylmethyl)thio]-6-arylthio DABO analogues. AB - 2-(Arylcarbonylmethyl)thio-6alpha-naphthylmethyl derivatives of dihydro-alkoxy benzyl-oxopyrimidines (DABO) were newly found to exhibit activity against both HIV-1 and HIV-2. To further explore their structure-activity relationship, the modified S-DABO analogues (5a-g and 6e-f) with a 1-naphthylthio or phenylthio group at the C-6 position were synthesized. S-Alkylation of 5-ethyl-2-thiouracil with substituted 2-bromo-acetophenones provided crude 2 [(arylcarbonylmethyl)thio]-5-ethyl-(3H)-uracil 2a-e, which was directly subjected to toluenesulfonylation with TsCl to afford disulfonate 4a-e. Substitution of 4a e with arylthiol afforded the desired S-DABO analogues 5a-g and 6e-f. The compounds were evaluated for their in vitro anti-HIV activity in MT-4 cells. The IC(50) values for anti-HIV-1 activity fall into the range 0.37-29.50 microM, and the IC(50) values for anti-HIV-2 activity fall into the range 23.11-181.07 microM. The results indicated that these compounds are moderately active against HIV-1 and HIV-2. PMID- 16211653 TI - Synthesis and antibacterial activity of some imidazole-5-(4H)one derivatives. AB - In the present study, several substituted oxazolones were synthesized by condensation of benzoylglycine with different aldehydes. From such oxazolones, substituted imidazolones were synthesized by condensation with ethylenediamine, urea and 4-N,N-dimethylaminoaniline. All these synthesized compounds produced significant antibacterial activities. Furthermore, compounds containing CH(2)CH(2)NH(2), -CONH(2) and -C(6)H(4)-N(CH(3))(2) groups as substitutents on the imidazolones were found to be potent antibacterial agents. Thus, among the twelve compounds, 1-(2-aminoethyl)-2-phen yl-4-(4 (dimethylamino)benzylidene)imidazole-5-(4H)one (4d), 1-carboxamido-2-phenyl-4-(4 (dimethylamino)benzylidene)imidazole-5-(4H)one (4e) and 1-(4-(N,N dimethylamino)phenyl)-2-phenyl-4-(4-(dimethylamino)benzylidene)imidazole-5 (4H)one (4f) were found to have a significant higher antibacterial activity than the other substituted imidazolones. Compound 4e was the most active one in this series. PMID- 16211655 TI - Adaptation of DELFIA cortisol kit for determination of salivary cortisol concentration. AB - Salivary cortisol concentration seems to be an excellent indicator of the biologically active plasma cortisol level because of the nearly absence of corticoid-binding proteins in saliva. With regard to the easy, noninvasive, and stress-free nature of saliva sampling, this parameter greatly facilitates studies of the HPA (hypothalamus-pituitary-adrenocortical) axis, especially in children. A commercially available TR-FIA (time-resolved fluoroimmunoassay), the DELFIA (dissociation-enhanced lanthanide fluoroimmunoassay) method, proposed for plasma and urine cortisol analyses, was adapted for salivary cortisol measurement by means of simple modifications of the assay protocol. The sensitivity was determined to be 0.53 nmol/L. The intra- and interassay coefficients of variation ranged from 5.5 to 8.2 % and from 6.0 to 10.4 %, respectively. The present findings suggest that the DELFIA procedure provides a reliable, sensitive, and convenient alternative procedure to the assay for salivary cortisol. PMID- 16211656 TI - Synthesis and biological evaluation of bis(methoxy methyl)-7,8-dihydro [1,4]dioxino[2,3-g]quinazolines as EGFR tyrosine kinase inhibitors. AB - A series of 7,8-bis(methoxymethyl)-7,8-dihydro-[1,4]dioxino[2,3-g]quinazolines were prepared and evaluated for their inhibition of phosphorylation of the isolated epidermal growth factor receptor (EGFR) enzyme and for their growth inhibition of the A431 cell line. Among them, compound 3c having a 3-iodophenyl ring was most potent (IC(50) = 1.66 nM) against the isolated EGFR enzyme and also showed meaningful potency (GI(50) = 1.99 microM) against the A431 cell line, although less than PD153035 (GI(50) = 1.03 microM). However, compound 3e as the exact rigidified analogue of Erlotinib (Tarceva) was inferior to the original compound when compared to its reported data. PMID- 16211657 TI - Development of albumin-binding doxorubicin prodrugs that are cleaved by prostate specific antigen. AB - Prostate-specific antigen (PSA) is a serine protease that is overexpressed in prostate carcinoma and represents a molecular target for selectively releasing an anticancer agent from a prodrug formulation. In this work, we developed albumin binding prodrugs with the structures MT-Ser-Ser-Tyr-Tyr- Ser-Gly-DOXO, MT-Asn-Ser Ser-Tyr-Phe-Gln-DOXO (MT = maleimidotriethyleneglycol acid; DOXO = Doxorubicin) or EMC-Arg-Arg-Ser-Ser-Tyr-Tyr-Ser-Gly-DOXO (EMC = epsilon-maleimidocaproic acid; X = amino acid). The maleimide Doxorubicin derivatives bound rapidly to the cysteine-34 position of endogenous and exogenous albumin and were efficiently cleaved by PSA at the P(1)-P'(1) scissile bond, releasing a respective Doxorubicin dipeptide (Ser-Gly-DOXO or Phe-Gln-DOXO). The derivative containing arginine residues (EMC-Arg-Arg-Ser-Ser-Tyr-Tyr-Ser-Gly-DOXO) exhibited excellent water solubility for intravenous administration. Subsequent biological evaluation was focused on a PSA-negative xenograft model (PC 3) and a PSA-positive xenograft model (CWR22) in order to assess the selectivity of our therapeutic approach. EMC Arg-Arg-Ser-Ser-Tyr-Tyr-Ser-Gly-DOXO showed no in vivo activity in the PSA negative PC 3 model, but good activity in the CWR22 PSA-positive model that was comparable to Doxorubicin. PMID- 16211658 TI - Determination of salbutamol in pharmaceuticals by capillary electrophoresis. AB - A capillary electrophoresis (CE) method for the separation and determination of salbutamol (SAL) in commercial pharmaceuticals was developed. Several parameters affecting the separations were studied including type and concentration of the carrier ion, counter ion, and the pH of the buffer. A hydrodynamically closed CE separation mode employing a 300 mum I.D. capillary, dynamically coated by polymeric buffer additive, provided good performance parameters (precision, accuracy, sensitivity). The obtained results suggest that the proposed method is suitable for routine assay of SAL in pharmaceutical preparations and can be used as an alternative to the conventional open CE mode for this application. PMID- 16211659 TI - N,N-dichlorotaurine: chemical and bactericidal properties. AB - The biogenous antimicrobial agent N-chlorotaurine (NCT) converts by disproportionation to N,N-dichlorotaurine (NDCT) at a rate proportional to acidity. This occurs at appreciable amounts already in weakly acidic biological systems. To understand the consequences of NDCT formation, a thorough investigation of this undescribed compound was mandatory, which needed its synthesis. Differently from NCT, this was possible in the aqueous system using trichloroisocyanuric acid. While the free acid, Cl(2)HNCH(2)CH(2)SO(3)H, was not available in pure form, its sodium and potassium salts were analytically pure and showed melting points (decomposition) of 125-128 degrees C (potassium) and 162 164 degrees C (sodium). The sodium salt demonstrated unexpected long-term stability even at room temperature (8.4 % loss of activity within 4 months). The aqueous solutions of both salts exhibited a weak acid reaction, and they were less stable than NCT. With regard to chlorination of amines (transhalogenation), NDCT was, surprisingly, less efficacious than NCT, which manifested itself by a lack of reactivity at pH < 7, for which a mechanistic explanation is given. Compared on a molar scale, NDCT was more bactericidal than NCT against the gram negative bacteria E. coli, P. aeruginosa and P. mirabilis, while there was no difference concerning the gram-positive ones, S. aureus and S. epidermidis. The increase of bactericidal activity at acidic pH was the same as observed with NCT and is attributed to a higher susceptibility of bacteria in this environment. Taken together, NDCT seems not to be suited to substitute NCT as a preparation fit for medical practice. PMID- 16211660 TI - Effect of pyrazoloquinoline derivatives on the growth of Leishmania donovani promastigotes. AB - A screening study was conducted to examine the effect of a series of synthesized pyrazoloquinoline derivatives on the growth of Leishmania donovani promastigotes. Sixteen compounds were tested, ten of which showed an inhibitory effect on the growth of promastigotes. Compound 1 demonstrated potent antileishmanial activity, followed by compounds 3 and 7. Some compounds showed less significant activities, while others exhibited little or no activity. Some of these compounds may be potential candidates for future treatment of leishmaniasis. PMID- 16211663 TI - Educational puzzles for understanding gastrointestinal physiology. AB - We developed four innovative, creative, and fun educational tools to promote active learning, enhance problem-solving skills, and encourage small group discussion. Furthermore, the tools encourage deductive reasoning and critical thinking rather than passive memorization of material. The tools include crossword puzzles, hidden messages, word scrambles, and word searches. These tools were developed using two computer programs: the Crossword Construction Kit and The New Puzzle Factory. Instructors are encouraged to optimize the value of the tools by using the additional options presented at the end of each of the puzzles. The additional options encourage students to become active learners by creating their own tools. Although the principles of these four tools can be adapted to many disciplines, these specific games focused on gastrointestinal physiology. Our goal was to create tools that can be used either inside or outside the classroom to complement and enhance the lecture. PMID- 16211661 TI - Mobility and leaching of glyphosate: a review. AB - There is currently concern that glyphosate, a strongly sorbing non-selective herbicide which is widely used in Europe, may be leached from the root zone into drainage water and groundwater. The purpose of this review is to present and discuss the state of knowledge with respect to the mobility and leaching of glyphosate from agricultural soils. Specific attention is given to the adsorption behaviour of glyphosate and the analysis of available studies on glyphosate transport. In addition, there are a number of experimental and numerical studies indicating that other strongly sorbing substances may be transported rapidly to the sub-surface. The experimental studies analysed in the paper encompass column , lysimeter- and field-scale experiments on glyphosate transport. The experimental findings, combined with transport studies on other strongly sorbing pesticides in the literature, support the hypothesis that transport of glyphosate may be caused by an interaction of high rainfall events shortly after application on wet soils showing the presence of preferential flow paths. Concentrations of glyphosate in European groundwater have been reported occasionally but monitoring is still limited. PMID- 16211664 TI - A laboratory demonstration for learning about mast cell degranulation. AB - A simple experimental model of cell degranulation was implemented that exposed mast cells obtained from Sprague-Dawley rats to saponin. The model is flexible, asy, and low cost, is not very time-consuming to run, and needs a minimum of laboratory resources. It has been used for the last three years in our undergraduate medical physiology courses and has replaced the classic utilization of slides and drawings. PMID- 16211665 TI - "Virtual rat": a tool for understanding hormonal regulation of gastrointestinal function. AB - This manuscript describes a "dry laboratory" using the "virtual rat" to help students understand the hormonal regulation of gastrointestinal function. The laboratory was modeled after a recent exercise that used the virtual rat to teach basic endocrine physiology. The virtual rat concept avoids the many obstacles associated with animal experimentation (for example, lack of adequate animal facilities, expense, equipment, and limited teacher experience). Our goal was to create a fun and educational experience while avoiding the complications associated with laboratory experimentation. No additional materials are required to complete this exercise. After finishing this laboratory, the students should have a greater understanding and appreciation for experimental design and the collection, analysis, and interpretation of data. PMID- 16211666 TI - Use of course-embedded projects for program assessment. AB - There is increasing demand on science faculty to develop authentic assessment measures for both individual courses and undergraduate programs. We report here on a quarter-long group project used in a neurophysiology course that can be used for either purpose. Small groups of four to five students critically analyze at least 10 articles from the primary scientific literature. The end result of this process is the equivalent of a scientific review article that is presented in two formats, a 10-min oral presentation and a scientific poster presentation. Students perform better on application tasks than on analysis, synthesis, or evaluation tasks associated with the project (P < 0.025) and generally respond positively to process questions (59-82%) but less positively to task questions (36-76%) about group dynamics. The cognitive skills and basic content knowledge required to complete this project are developed throughout the undergraduate program. Thus the project is a type of culminating program experience. However, the project also assesses basic course proficiency, because students cannot analyze primary neuroscience research without an understanding of neurophysiological principles. PMID- 16211667 TI - Telemedical education: teaching spirometry on the Internet. AB - Advances in portable equipment have led to routine spirometry testing outside of formal pulmonary function laboratories. Practitioners ordering these tests are not formally trained in spirometry interpretation. Providing effective off-site training can be challenging. Our objective was to develop a remotely accessible computer-based tutorial for teaching spirometry interpretation to nonpulmonologists. We designed an educational module that was accessible via the Internet and tested by 65 medical trainees at a major university medical center. In addition, the module was posted within the Virtual Hospital on the World Wide Web. Increases in spirometry interpretative skills were assessed using pre- and post-tests submitted electronically. The spirometry module significantly improved spirometry interpretation by nonspecialist trainees. This improvement included a broad increase in knowledge base and was observed independent of training level and prior spirometry reading experience. We conclude that computer-based tutorials can effectively train off-site practitioners in spirometry interpretation. This technology allows for the dissemination of educational material from a central site of expertise and provides a valuable adjunct to limited teaching resources. PMID- 16211668 TI - Helping students to understand that outward currents depolarize cells. AB - The physiology of excitable membranes is a fundamental topic in neuroscience and physiology courses at graduate and undergraduate levels. From the building blocks of ionic gradients and membrane channels whose permeability is selective and variable, we build the concepts of resting potential, action potential, and propagation in neurons and muscle fibers. Many students have an intuitive understanding of the movements of ions and the associated changes in membrane potential. For example, potassium ions leaving a cell through potassium-selective channels become unbalanced positive charges on the outside of the cell (and leave unbalanced negative charges on the inside), thus producing a potential across the membrane with the inside negative with respect to the outside. Later, when we discuss the local circuit currents that underlie propagation or the basis for extracellular stimulation, we make the general statement that "outward currents depolarize cells." Students respond with utter disbelief. Two simple additions to a discussion of membranes are suggested that permit the formulation of a consistent set of rules that apply to everything from the resting and action potentials of nerve and muscle through synaptic potentials and stimulation techniques. PMID- 16211669 TI - The beaver pond analogy of blood glucose control. AB - The concentration of glucose is tightly regulated in the blood by a complicated set of physiological variables. To provide students with a means to more readily understand these complex mechanisms, the control of the water level in a beaver pond is presented as an analogy. A beaver must maintain a constant water level in the pond for the proper functioning of the lodge, just as blood glucose is maintained for, among other reasons, brain function. The beaver controls the water level by changing outflow over the dam and inflow from stream beds. Water flow over the dam is analogous to glucose leaving the blood for tissues, which is controlled by insulin. Inflow of water from streams is analogous to glucose absorption from the gastrointestinal tract and glucose release from the liver, the latter being controlled by glucagon and other counterregulatory hormones. The analogy is extended by considering the effects of exercise in normal and diabetic individuals on blood glucose levels. PMID- 16211670 TI - Case-stimulated learning within endocrine physiology lectures: an approach applicable to other disciplines. AB - The problem of presenting endocrine physiology lectures in a format that interests medical students was addressed. Incorporating truncated case-stimulated learning sessions into the lectures has proven to be a successful solution to this issue, while also providing continuity between topics in the lecture block. This method of providing a direct clinical link for basic physiological concepts and providing a more active learning experience is adaptable to most basic science disciplines. PMID- 16211671 TI - Unilateral nephrectomy in the rat as a teaching model of renal function. AB - The white rat offers a valuable teaching model to demonstrate the in situ anatomy, homeostatic control, and function of the mammalian renal system. Students in the upper-level physiology class at Dickinson College are responsible for formulating a set of testable hypotheses as to the possible compensatory mechanisms that will result from a unilateral nephrectomy in the rat. To test their hypotheses, each group of students is made responsible for induction and maintenance of anesthesia, removal of the left kidney, closure, and recovery of the animal in their experiment. Before and 1 wk after nephrectomy, blood samples are collected via tail section for measurement of hematocrit and plasma creatinine concentrations. Pooling data across all student groups (n = 6) for pre and postnephrectomy hematocrit and creatinine values allows students to run statistical comparisons testing their hypotheses. This laboratory experience in integrative physiology is a successful exercise in introducing fundamentals of experimental design and, most importantly, provides students with an appreciation for the process of humane use of animals in research and teaching. PMID- 16211672 TI - Peer tutoring improves student performance in an advanced physiology course. AB - Effectiveness of a peer tutoring system developed for an advanced physiology course was assessed in terms of academic performance and perceived value. Forty five students took the course without the peer tutoring system, and sixty-nine students took the course with peer tutoring. Grades from both groups of students were compared with grades earned in an introductory physiology course. Tutored students were asked how much they valued the peer tutoring. There was a decline in grades received by the students in the advanced physiology course compared with their scores in the introductory physiology course in both tutored and untutored groups. However, the decline in the tutored group was significantly (P = 0.015) less than that in the untutored group of students. Tutored students reacted very favorably to the tutoring sessions and expressed a desire to see tutoring expanded to other courses. This was the first demonstration of the effectiveness of peer tutoring in college science teaching. Peer tutoring appears to be effective in enhancing student performance as well as being perceived as beneficial by the students. PMID- 16211673 TI - How accurate are our assumptions about our students' background knowledge? AB - Teachers establish prerequisites that students must meet before they are permitted to enter their courses. It is expected that having these prerequisites will provide students with the knowledge and skills they will need to successfully learn the course content. Also, the material that the students are expected to have previously learned need not be included in a course. We wanted to determine how accurate instructors' understanding of their students background knowledge actually was. To do this, we wrote a set of multiple-choice questions that could be used to test students' knowledge of concepts deemed to be essential for learning respiratory physiology. Instructors then selected 10 of these questions to be used as a prerequisite knowledge test. The instructors also predicted the performance they expected from the students on each of the questions they had selected. The resulting tests were administered in the first week of each of seven courses. The results of this study demonstrate that instructors are poor judges of what beginning students know. Instructors tended to both underestimate and overestimate students' knowledge by large margins on individual questions. Although on the average they tended to underestimate students' factual knowledge, they overestimated the students' abilities to apply this knowledge. Hence, the validity of decisions that instructors make, predicated on the basis of their students having the prerequisite knowledge that they expect, is open to question. PMID- 16211674 TI - Whither the art of teaching? PMID- 16211675 TI - Report from Russia. PMID- 16211676 TI - Stem cells may protect brain, nervous system. PMID- 16211677 TI - HRT not recommended for prevention of chronic diseases. PMID- 16211679 TI - More warnings about common pain relievers. PMID- 16211678 TI - Rating your hospital. PMID- 16211680 TI - Acupuncture aids overactive bladder. PMID- 16211681 TI - Aerobic exercise can improve sexual function. PMID- 16211682 TI - Bortezomib increases survival of patients with relapsed multiple myeloma. PMID- 16211683 TI - Men in relationships fare better after prostate cancer treatment. PMID- 16211684 TI - Rx: 10,000 daily steps. PMID- 16211685 TI - Depression, pain linked. PMID- 16211686 TI - I've heard a lot about the "microfracture" technique for the repair of damaged knee cartilage, specifically that it is very effective and lasts a long time. Can you explain the procedure and what its advantages are over conventional methods of knee repair? PMID- 16211687 TI - I have gout, and my doctor told me not to take aspirin. Why? PMID- 16211688 TI - Felon's DNA, HIV testing order upheld by appeals court. PMID- 16211689 TI - Global courts. South African judge jails HIV-positive rapist for life. PMID- 16211690 TI - Filing. HIV patient denied treatment sues doctors. PMID- 16211691 TI - Taking quackery out of conservation. PMID- 16211692 TI - Efficacy of granulocyte-macrophage colony-stimulating factor on the survival of ischaemic skin flaps in rats. AB - Our objective was to find out whether granulocyte-macrophage colony-stimulating factor (GM-CSF) improves the survival of skin flaps in rats. Random skin flaps were raised in 30 male Sprague-Dawley rats, and recombinant GM-CSF 50 microg in 0.5 ml saline solution (n=15) or 0.5 ml saline solution alone (n=15) was injected intradermally into the area of the flap after it had been raised. Surviving areas were measured after seven days, and the median area surviving in the group treated with GM-CSF was greater than that in the control group (728 mm2 (range 565-1120) compared with 985 mm2 (range 636-1368), p=0.005). These results suggest that in this model, GM-CSF given postoperatively improves the viability of ischaemic skin flaps. PMID- 16211693 TI - Functional outcome after surgical treatment of phalangeal fractures in severely injured hands. AB - We assessed functional results after treatment of phalangeal fractures in severely injured hands. Our aim was to quantify digital functional loss with (combinations of) risk factors of unsatisfactory function. Patients who had multiple phalangeal fractures necessitating operation in a 10-year time period were tested, using measurements of total active movement. Seventy-eight patients with 228 phalangeal fractures were available for follow-up. In 88 fingers, the fractures ended in amputation and were excluded from the study. In the resulting 140 fractures, 74 (53%) had a good result (movement >180degrees for fingers 2-5, and >98degrees for the thumb), and 66 (47%) in an unsatisfactory result. Associated soft tissue injury, level of injury, and arthrodesis were risk factors for diminished function. Intra-articular fractures and multiple fractures within the same finger predisposed to arthrodesis. Despite the extensive and severe injuries more than half had good results, which is comparable with reports describing hand injuries with less extensive trauma. PMID- 16211694 TI - Scaphotrapeziotrapezoid arthrodesis for the treatment of Lichtman stage 3B Kienbock disease. AB - We evaluated the results clinically and radiologically of 10 cases of Lichtman stage 3B Kienbock disease treated by scaphotrapeziotrapezoid (STT) arthrodesis alone. We retrospectively reviewed pain, range of movement, grip strength, and physical examination of the wrist as well as subjective satisfaction. We measured carpal height ratio and radioscaphoid angle. The median follow-up period was 53 months (range 10-109). At final evaluation, eight patients reported no pain and two had mild pain. Median range of movement was 60degrees(range 45-70degrees) in extension, 33degrees (range 20-45degrees) in flexion without appreciable loss. Grip strength had improved considerably. Employed patients went back to work within four months. The median final radioscaphoid angle was 67degrees (range 55 81degrees). STT arthrodesis done properly for the right indications for Lichtman stage 3B Kienbock disease achieves quick relief of pain and excellent function of the wrist. PMID- 16211695 TI - Nodular fasciitis of the ulnar nerve at the palm. AB - A 42-year-old woman complained of progressive induration in the right palm. As the mass was impossible to separate from the ulnar nerve, we excised the mass together with the digital nerve and grafted 4cm of the sural nerve. The final diagnosis was nodular fasciitis. PMID- 16211696 TI - Re: Laterally moved, coronally advanced flap: a modified surgical approach for isolated recession-type defects. PMID- 16211698 TI - Mary P. Edmonds (1922-2005). PMID- 16211697 TI - Relief of vasomotor symptoms and vaginal atrophy with three doses of conjugated estrogens and medroxyprogesterone acetate in postmenopausal Asian women from 11 countries: The Pan-Asia menopause (PAM) study. AB - OBJECTIVE: To evaluate the efficacy of three doses of estrogen/progestin therapy for relief of vasomotor symptoms (VMS) and vaginal atrophy in Asian women of different ethnic background; to examine differences in prevalence of VMS among ethnic groups. METHODS: A prospective, randomized, double-blind multinational clinical trial in healthy postmenopausal women from 11 Asian countries. Following 2 weeks of baseline observations, the women received one of three conjugated estrogens (CE)/medroxyprogesterone acetate (MPA) doses (in mg) daily for 24 weeks: 0.625/2.5; 0.45/1.5; or 0.3/1.5. The women recorded VMS and uterine bleeding daily on diary cards translated into 10 languages. Vaginal responsiveness was evaluated by the vaginal maturation index (VMI) at baseline and at week 24. RESULTS: The study population consisted of 1028 postmenopausal women. The VMS-evaluable subpopulation was about 60% of the total population. The mean baseline hot flush frequency was 1.6 flushes/day (613 women). Hot flush frequency decreased significantly in all dose groups within 4 weeks of treatment. The VMI shifted significantly from immature (parabasal) to mature (superficial) cells at end of treatment. The therapeutic responses were comparable in all three groups. However, uterine bleeding was consistently less frequent in the 0.3/1.5 mg group. The percentage of women who reported VMS at baseline differed substantially among the different ethnic groups, ranging from 5% in Indonesian women to 100% in Vietnamese women. CONCLUSION: Asian postmenopausal women respond to CE/MPA therapy. The lowest dose is as effective for VMS and vaginal responsiveness as the higher doses, and the lowest dose is associated with the most favorable bleeding pattern. The prevalence of vasomotor symptoms differs among ethnic groups. PMID- 16211699 TI - Prevalence of endoscopically negative and positive gastroesophageal reflux disease in the Japanese. AB - OBJECTIVE: The frequency of gastroesophageal reflux disease (GERD) has not been fully investigated in the Asian population. The aim of this study was to investigate the prevalence of GERD, endoscopy-negative GERD (NERD), and erosive GERD in Japan, and the factors influencing disease prevalence. MATERIALS AND METHODS: A total of 2760 subjects (mean age 50.4 years, range 24-84 years) were prospectively enrolled in this multicenter study. GERD symptoms were assessed with the Japanese version of the Carlsson-Dent self-administered questionnaire (QUEST) and upper gastrointestinal endoscopy was performed on all study participants. RESULTS: A total of 495 (17.9%) individuals were diagnosed with GERD by the presence of erosive esophagitis at endoscopy and/or by the presence of GERD symptoms. Erosive esophagitis was diagnosed endoscopically in 195 (7.1%), and symptomatic GERD was diagnosed in 351 (12.7%) based on a QUEST score of over 6. Of these 351 subjects, 300 (10.9%) were considered to have NERD. Male gender, hiatal hernia, and mild gastric mucosal atrophy were significant positive predictive factors of erosive esophagitis by multiple regression analysis. Hiatal hernia ws the only significant predictor of GERD symptoms. Traditional Japanese foods, such as sweet cakes and rice cake, frequently exacerbated GERD symptoms. CONCLUSIONS: The prevalence of GERD in the Japanese was 17.9% and the prevalence rates of NERD and erosive esophagitis were 10.9% and 8.6%, respectively. The majority of symptomatic patients did not have endoscopically proven esophagitis. Hiatal hernia is the only important predictor of the presence of GERD symptoms. PMID- 16211700 TI - Saliva transport to the distal esophagus. AB - OBJECTIVE: During esophageal acid clearance, saliva should reach the most distal esophagus. The mechanisms responsible for saliva transport are not completely understood but it is assumed that normal peristalsis plays a significant role. The aim of this study was to assess the role of esophageal peristalsis and gravity in saliva transport to the distal esophagus. MATERIAL AND METHODS: Esophageal transit and presence times of a 2-ml bolus of radiolabeled artificial saliva were assessed using concurrent scintigraphy and manometry in 10 healthy volunteers in the upright and supine positions before and after disruption of esophageal motility with sildenafil (50 mg). RESULTS: With normal peristalsis, there was no difference in saliva transit to the distal esophagus between supine and upright positions 3.9 (1.5- >60.0) versus 3.3 s (1.3-8.3). Low amplitude contractions did not affect saliva transit but the disappearance of contractions after sildenafil was associated with prolonged saliva transit in supine position 7.4 (1.0- >60.0). Saliva presence time was significantly prolonged in both the upright and supine positions by esophageal dysmotility. CONCLUSIONS: Saliva transport to the distal esophagus does not require complete normal peristalsis or gravity and mainly depends on an efficient pharyngeal pump. However, subjects in supine position with severe esophageal dysmotility might have both impaired volume clearance and delayed saliva transport, leading to abnormal acid clearance and esophagitis. PMID- 16211701 TI - Seroconversion of Helicobacter pylori in Korean male employees. AB - OBJECTIVE: Helicobacter pylori infections are acquired primarily during childhood, but also occur in adults. As no published estimates are available on the seroconversion rate of H. pylori in the Korean population, the purpose of this study was to evaluate the incidence and prevalence of H. pylori infection in male Korean employees. MATERIAL AND METHODS: Levels of anti-H. pylori IgG were measured by ELISA, and gastroscopic findings were checked in 588 employees of the Subway Corporation (aged 40-60 years, male) in 1997, 1998, 1999, 2000, and 2003. Questionnaire responses concerning socio-economic state, education level, and type of job were analyzed versus serologic test results. RESULTS: Out of the 588 subjects, 455 (77.4%) were positive for anti-H. pylori IgG. Multivariate analysis showed that abnormal gastroscopic findings, especially duodenal ulcers, were more frequent in the H. pylori-positive group (17.6) than in the initially H. pylori negative group (9.0%) (OR: 1.71, 95% CI: 1.07-2.81). Among the multiple questionnaires, only the proportion of white-collar workers was significant; that is, it was lower in the H. pylori-positive group (44/6) than in the initial H. pylori-negative group (57.9%) (OR: 0.62, 95% CI: 0.41-0.92). Seventeen of 133 subjects (12.8%) who were initially seronegative underwent seroconversion during a 6-year follow-up period, a seroconversion rate of 2.13%/annum. CONCLUSIONS: The seroprevalence of H. pylori infection in this study group was 77.4% and duodenal ulcer was more frequent in H. pylori-positive subjects, thus confirming the relationship between H. pylori infection and duodenal ulcer. Seroconversion of H. pylori infection in the Korean male employees was determined to be 2.13% per year, suggesting that new infections occur continuously even during late adulthood. PMID- 16211702 TI - HIV-1. Viral assembly inhibitors on the horizon. PMID- 16211704 TI - Bush faces stem cell pressure. PMID- 16211703 TI - Processes of change in brain and cognitive development. Proceedings of the XXIst Meeting of the International Association for the Study of Attention and Performance. July 26-August 1, 2004. Winter Park, Colorado, USA. PMID- 16211706 TI - V.S. Ramachandran. PMID- 16211705 TI - Island revelations. PMID- 16211707 TI - Nesiritide--not verified. PMID- 16211708 TI - Geranylgeranylacetone protects mice from dextran sulfate sodium-induced colitis. AB - OBJECTIVE: Geranylgeranylacetone (GGA) has recently been reported to induce heat shock protein (HSP) 70, which has a protective function against inflammation. We investigated the therapeutic effects of oral administration of GGA on dextran sulfate sodium (DSS)-induced colitis in mice. MATERIALS AND METHODS: BALB/c mice were given 3% DSS solution orally for 7 days to induce colitis. The disease activity of colitis was assessed clinically every day, and histology in the colon was evaluated at 7 days post-DSS. The levels of myeloperoxidase (MPO) activity, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma in the colon tissues were also examined. In addition, expression of HSPs 25, 40, 70 and 90 in the colon tissue was determined by Western blot analysis. Mice were orally administered GGA (50-500 mg/kg) when treatment of DSS started. RESULTS: It was found that GGA significantly reduced the clinical severity of colitis and suppressed the levels of MPO activity, TNF-alpha and IFN-gamma induced by DSS in the colon. On the other hand, GGA enhanced the expression of HSP70 in the colon of mice given DSS. CONCLUSIONS: Taken together, these results suggest that GGA is a new anti-inflammatory drug that could be useful in the treatment of colitis such as inflammatory bowel disease. PMID- 16211709 TI - Nesiritide--not verified. PMID- 16211710 TI - Torcetrapib and atorvastatin. PMID- 16211711 TI - Torcetrapib and atorvastatin. PMID- 16211712 TI - Case 17-2005: acute chest syndrome and ARDS. PMID- 16211713 TI - Expression of keratinocyte growth factor and its receptor in adaptive changes of ileorectal pouch mucosa. AB - OBJECTIVE: Total proctocolectomy with formation of an ileo-anal pouch is a well established surgical procedure for patients with ulcerative colitis (UC) or familiar adenomatous polyposis (FAP). The pouch mucosa undergoes adaptive changes, with inflammation of the ileal reservoir being the most common complication. The aetiology is unknown. The keratinocyte growth factor (KGF) has not only been shown to promote intestinal wound healing and re-epithelialization but also to have some immunomodulatory properties. This study was designed to investigate a putative involvement of KGF in observed histomorphological changes in the pouch mucosa. MATERIALS AND METHODS: Multiple biopsies were obtained from age-matched and sex-matched patients. Biopsies were stained with H&E and scored for inflammation and morphological changes. mRNA expression levels of KGF and KGF receptor (KGFR) were determined using competitive RT-PCR, protein expression and phosphorylation was analyzed by Western blotting. KGF and KGFR were localized in tissue samples by immunohistochemistry. RESULTS: Expression of KGF and KGFR was significantly increased in inflamed and adapting mucosa. Patterns of expression did not significantly differ in pouch mucosa from UC or FAP patients. Protein expression correlated with the mRNA results and KGFR was shown to be activated in adapting pouch mucosa. KGF was detected on subepithelial cells, mainly on fibroblasts, whereas expression of KGFR was restricted to epithelial cells. CONCLUSION: Expression of KGF and KGFR is significantly increased in the pouch mucosa, suggesting an involvement of this growth factor in tissue repair and adaptive changes. Topical application of KGF might alleviate the inflammatory and promote the adaptive process in the ileo-anal pouch mucosa. PMID- 16211714 TI - Efficiency of colorectal cancer surveillance in patients with ulcerative colitis: 26 years' experience in a patient cohort from a defined population area. AB - OBJECTIVE: Patients with ulcerative colitis (UC) have an increased risk of developing colorectal cancer (CRC). The current procedure to diminish this risk is colonoscopic surveillance and histopathological evaluation of biopsy specimens. This method is not unquestioned and is undergoing continuous evaluation. The present study is a major update of an earlier reported investigation from an ongoing surveillance programme. MATERIAL AND METHODS: In 1977 a colonoscopic surveillance programme comprising all patients with UC from a defined area was started in Ornskoldsvik. Three principal investigators performed regular colonoscopy with mucosal sampling for histopathological evaluation. Some 211 patients were studied from 1977 to 2002. At the end of the study period, 90 patients, including those operated on, had total colitis (TC) for more than 10 years, 74 patients had left the study, 31 after panproctocolectomy (PPC), 6 owing to advanced biological age, 1 because of intercurrent disease, 23 patients had moved out of the area and 13 patients were excluded because of poor compliance. In all, 928 colonoscopies were performed. RESULTS: It was found that 135 patients had radiologically or morphologically defined TC and 69 patients had left-sided colitis (LC). Nine CRCs were diagnosed in 8 patients, one of whom died of CRC, while another two were included in the programme with a diagnosis of CRC. Morphological alterations classified as dysplasia or indefinite for dysplasia (ID) were found in 52 patients, 5 of whom were later found to have CRC. Eighteen of the patients were operated on for different kinds of colonic resections and in 31 patients a PPC was performed. CONCLUSIONS: Colonoscopic surveillance is an effective method in preventing death from CRC among patients with UC. A conservative approach to surgery seems to be justified. The burden of the surveillance programme has been acceptable. The outcome depends on good patient compliance as well as the involvement of as few investigators as possible. PMID- 16211715 TI - Survival following the development of ascites and/or peripheral oedema in primary biliary cirrhosis: a staged prognostic model. AB - OBJECTIVE: Current prognostic models in primary biliary cirrhosis (PBC) have low precision, partly due to the restricted inclusion criteria of some cohorts used for modelling but also because of the prolonged natural course of the disease. It is hypothesized that better precision could be achieved with a staged model, using ascites or peripheral oedema as a new starting-point for prediction. MATERIAL AND METHODS: The study was based on an established database of 289 consecutive patients, followed between 1977 and 1998. Stepwise Cox regression was used to construct a staged model based on 143 patients who first developed ascites (n=111) or peripheral oedema (n=32) at entry or during subsequent follow up. The model was compared with published models using graphical methods and receiver operating characteristics (ROCs). RESULTS: Mean time from clinical diagnosis of ascites or peripheral oedema to death was 3.1 years. The following variables had independent prognostic significance: log10(bilirubin) (p<0.001), albumin (p<0.001), age (p<0.001) and history of encephalopathy (p<0.001). Goodness of fit showed that the survival probabilities predicted by the Ascites Stage Model fitted well with the observed data. The Ascites Stage Model (ROC 0.8324 (SE 0.0348)) was a better predictor of survival than the Mayo long-term model (ROC 0.7833 (SE 0.0397)), the Mayo Repeated Patient Visits Model (ROC 0.7779 (SE 0.0399)) and the Royal Free PBC Prognostic Model (ROC 0.7785 (SE 0.0396)). CONCLUSIONS: The Ascites Stage Model gives a better survival estimate for PBC patients once they have developed ascites or peripheral oedema compared with the current models, and demonstrates an advantage of staged models in diseases with a prolonged natural history. PMID- 16211716 TI - Intestinal permeability and bacterial growth of the small bowel in patients with primary sclerosing cholangitis. AB - OBJECTIVE: Animal studies show that small intestinal bacterial overgrowth and infusion of bacterial antigens into portal blood cause hepatic histological changes similar to those seen in primary sclerosing cholangitis in man. It has been suggested that aa similar mechanism involving bacterial overgrowth with increased small-bowel permeability may play a pathogenic role in patients with primary sclerosing cholangitis (13 M, 9 F, median age 37 years, range 21-74 years), 19 of whom (83%) had quiescent inflammatory bowel disease, were included in the study along with 18 healthy volunteers (9 F, ( M, median age 36 years, range 23-80 years). Small-bowel bacterial overgrowth was defined as the presence of colonic flora>10(5) colony-forming units (cfu)/ml from duodenal aspirations. Small-bowel intestinal permeability was assessed as the differential urinary excretion of lactulose/L-rhamnose. RESULTS: Bacterial overgrowth was evident in one patient with primary sclerosing cholangitis (4.5%) (Enterobacter) and in none of the controls. Intestinal permeability in patients with primary sclerosing cholangitis (0.034 (0.026-0.041) (median, interquartile range (IQR)) did not differ significantly from that of the controls (0.033 (0.025-0.041). CONCLUSIONS: Small intestinal bacterial overgrowth and increased intestinal permeability does not seem to play an important pathogenic role in patients with primary sclerosing cholangitis. PMID- 16211717 TI - Diabetes is epidemic. PMID- 16211718 TI - Prevention of postoperative peritoneal adhesions: effects of lysozyme, polylysine and polyglutamate versus hyaluronic acid. AB - OBJECTIVE: Intraperitoneal adhesions are an important cause of postoperative intestinal obstruction, abdominal discomfort and infertility. In the present study we hypothesized that a combination of polypeptides with different surface properties, resulting in fine disperse low-soluble complexes, could be of benefit in the prevention of abdominal adhesions. MATERIAL AND METHODS: Various polypeptides including lysozyme, polyglutamate, polylysine and combinations of all three were evaluated as compared to hyaluronic acid. A standard wound on the parietal peritoneum in mice was used and the evaluated agents were administered immediately postoperatively. The extent of peritoneal adhesions to the injured area was measured and expressed as a percentage of the wound length as evaluated after 7 days. Flow cytometry was performed to evaluate the effect on peritoneal macrophage survival and phagocytic function and the Pick test was used to determine peroxide production in order to estimate toxicity and potential impairment of macrophage function caused by the chemicals. RESULTS: Significant differences were seen among the treatment groups (p<0.001). Both polyglutamate and lysozyme, and polyglutamate together with polylysine significantly decreased adhesion formation as compared to hyaluronic acid. The polylysine-polyglutamate combination was still visible macroscopically on the peritoneal surface after 1 week, though not after 1 month. The polyglutamate-lysozyme mixture was less effective than these individual components alone. The chemicals did not show any toxic effects or altered function in macrophage cell culture. CONCLUSIONS: Lysozyme, polyglutamate and, most effectively, a polyglutamate-polylysine combination significantly decreased experimental abdominal adhesion formation. A strong mechanical connection to the wound and prolonged attendance in the surface were noted. Peritoneal phagocyte function did not seem to be influenced by the chemicals. PMID- 16211720 TI - A family history of Barrett's oesophagus: another risk factor? AB - Barrett's oesophagus and oesophageal adenocarcinoma, although increasingly common, have no known genetic cause. In this report we describe a family with a remarkable history of Barrett's oesophagus and adenocarcinoma. The index case is a 76-year-old man with adenocarcinoma arising within Barrett's oesophagus. Two of his three brothers, aged 68 and 78 years, also developed adenocarcinoma arising in Barrett's oesophagus and the remaining 67-year-old brother has severe dysplasia in biopsies from Barrett's oesophagus. The sons and daughters of the index case requested screening and all had histologically confirmed short-segment Barrett's oesophagus. This kindred appears to be genetically susceptible to Barrett's oesophagus and oesophageal adenocarcinoma. Pooling of data from this and other Barrett's families may allow successful linkage analysis. PMID- 16211719 TI - Successful photo-and phenobarbital therapy during pregnancy in a woman with Crigler-Najjar syndrome type II. AB - Severe hyperbilirubinemia in a pregnant mother suffering from Crigler-Najjar syndrome type II is a threat to the unborn child and may result in brain injury. We report the case of a Gly71-->Arg/Tyr486-->Asp homozygous mother of East Asian descent, who was treated with phototherapy during embryogenesis and with phenobarbital during the rest of the pregnancy. This resulted in significantly reduced bilirubin levels in the mother, who gave birth to a healthy boy. A neonatal hyperbilirubinemia resolved spontaneously. PMID- 16211721 TI - Gastric duplication cyst: a rare endosonographic finding in an adult. AB - Cystic lesions located in the gastric wall are a rare finding in endosonography of the gastrointestinal tract. Compared to all forms of benign and malignant tumours of the stomach, gastric duplication cysts are an uncommon anomaly- especially in adults. We report on a 59-year-old woman suffering from intermittent abdominal pain, weight loss and nausea. A gastric duplication cyst was identified by endoscopic ultrasound (EUS), but malignancy was excluded by EUS and fine-needle aspiration histology. Because of continuously increasing abdominal complaints, surgery was performed with partial resection of the gastric corpus and splenectomy. Gross anatomy and histology showed a gastric cyst measuring 150 mm in maximum diameter with no evidence of malignancy or inflammation. Following surgery, the patient's condition recovered fully. PMID- 16211722 TI - Check your blood pressure at home. For a more accurate measure of your blood pressure, and help keeping it under control, try keeping tabs on it at home. PMID- 16211723 TI - Using Crestor--and all statins--safely. Some simple steps can help minimize or avoid muscle problems from these cholesterol-lowering drugs. PMID- 16211724 TI - No clear winner in traditional vs. off-pump bypass surgery. A panel of experts commissioned by the American Heart Association says that on-pump and off-pump bypass surgery are equivalent, though each has its advantages and disadvantages. PMID- 16211725 TI - Statins don't stop aortic valve narrowing...or memory loss (yet). PMID- 16211726 TI - No one-size-fits-all for aspirin and elders. PMID- 16211727 TI - Ask the doctor. What should you do if your pacemaker or implanted defibrillator is recalled? PMID- 16211728 TI - [55th general meeting of the Japanese Society of Allergology. October 20-22, 2005. Morioka, Japan. Abstracts]. PMID- 16211729 TI - Is tax reform a solution to the uninsured crisis in America? PMID- 16211730 TI - Negotiating ethics in anaethesia. PMID- 16211731 TI - Impact of the Narcotrend Index on propofol consumption and emergence times during total intravenous anaesthesia with propofol and remifentanil in children: a clinical utility study. AB - BACKGROUND AND OBJECTIVE: The electroencephalographic Narcotrend Index (NI) is a measure of the hypnotic component of general anaesthesia. The purpose of this study was to evaluate the impact of Narcotrend guidance on propofol consumption and emergence times in children receiving total intravenous anaesthesia with propofol and remifentanil. METHODS: Thirty children, aged 1-11 yr, scheduled for paediatric urological surgery were enrolled. Remifentanil was given to all patients at a constant infusion rate of 0.3 microg kg [-1] min[-1] throughout anaesthesia. Patients were randomly allocated to receive a continuous propofol infusion adjusted either according to a conventional clinical practice (Group C: n=15) or guided by Narcotrend monitoring (Group NI: n=15; target NI 60+/-5). All patients were connected to the Narcotrend Monitor, but in Group C the anaesthetist was blinded to the screen of the monitor. Propofol consumption (mg kg[-1]h[-1]) and emergence times (min) were the primary and secondary outcome measures. RESULTS: Propofol consumption (median [inter-quartile range]) was significantly lower in Group NI compared to Group C (NI: 7.0 [6.4--8.2] vs. C: 9.3 [8.3--11.0] mg kg[-1]h[-1]; P<0.001), whereas Log-Rank-analysis revealed no intergroup difference in emergence times (Group NI: mean [95% confidence interval (CI)] 12.8 [11.2--14.4] min; Group C: 16.4 [12.6--20.2] min; P=0.10). Haemodynamic variables remained stable within age-related limits, and there were no observations of adverse events, especially no clinical signs of intraoperative awareness in any patient. CONCLUSION: Narcotrend monitoring for guidance of propofol/remifentanil anaesthesia in children results in reduced propofol consumption compared to a conventional clinical practice. PMID- 16211732 TI - Intubation conditions after rocuronium or succinylcholine for rapid sequence induction with alfentanil and propofol in the emergency patient. AB - BACKGROUND AND OBJECTIVE: Previous studies mainly conducted on elective patients recommend doses of 0.9-1.2 mg kg[-1] rocuronium to obtain comparable intubation conditions with succinylcholine 1.0 mg kg[-1] after 60 s during a rapid-sequence induction. We decided to compare the overall intubating conditions of standard doses of rocuronium 0.6 mg kg[-1] and succinylcholine 1.0 mg kg[-1] during a strict rapid-sequence induction regimen including propofol and alfentanil. METHODS: Male and female patients (ASA I-III) older than 17 yr scheduled for emergency abdominal or gynaecological surgery and with increased risk of pulmonary aspiration of gastric content were randomized to a rapid-sequence induction with succinylcholine 1.0 mg kg[-1] or rocuronium 0.6 mg kg[-1]. Patients with a predicted difficult airway were excluded. A senior anaesthesiologist 'blinded' for the randomization performed the intubation 60 s after injection of the neuromuscular blocker. Intubating conditions were evaluated according to an established guideline. Tracheal intubation not completed within 30 s was recorded as failed. RESULTS: 222 patients were randomized. Three patients had their operation cancelled and 10 did not fulfil the inclusion criteria. Clinically acceptable intubation conditions were present in 93.5% and 96.1% of patients in the succinylcholine group (n=107) and the rocuronium group (n=102), respectively (P=0.59). CONCLUSIONS: During a rapid sequence induction with alfentanil and propofol, both rocuronium 0.6 mg kg[-1] and succinylcholine 1.0 mg kg[-1] provide clinically acceptable intubation conditions in 60 s in patients scheduled for emergency surgery. Under the conditions of this rapid-sequence induction regimen rocuronium may be a substitute for succinylcholine. PMID- 16211733 TI - Multiple-trauma management: standardized evaluation of the subjective experience of involved team members. AB - BACKGROUND AND OBJECTIVE: Staff attitude plays a pivotal role in quality management. The objective of the present study was to further define how interdisciplinary emergency hospital staff experience their daily work and the extent to which the professional speciality and training of an individual influences his/her assessment of multiple-trauma team performance. METHODS: The clinical staff involved in multiple-trauma emergency management of a university hospital was asked to answer a confidential questionnaire. Factorial analysis was used to identify 8 major dimensions from a total of 53 items. RESULTS: The questionnaire was returned by 128 team members. All professional groups were most dissatisfied with the dimensions 'education and training', 'work sequence between specialities' and 'communication between specialities'. Assessment of the quality of in-hospital emergency-trauma management differed significantly between professional specialities (ANOVA, F=5.2; P=0.028); surgeons gave the highest ratings for all but one dimension. Having taken an Advanced Trauma Life Support (ATLS) course influenced significantly the total rating of multiple-trauma treatments of anaesthetists and surgeons (F=5.5; P=0.024). CONCLUSIONS: The perceptions of interdisciplinary trauma team members without the completion of an ATLS training course were that they did not communicate enough with each other and that there were differences between their expectations and reality. The differences and the communication deficits were overcome in team members who had passed an ATLS course. PMID- 16211734 TI - Medical experimentation concerning chemical and biological weapons for mass destruction. AB - This article is the text of a speech originally presented at the Second World Conference on Medical Ethics at Gijon, Spain, on 2 October 2002 under the title "Medical Experimentation Concerning Chemical and Biological Weapons for Mass Destruction: Clinical Design for New Smallpox Vaccines: Ethical and Legal Aspects." Experimentation on vaccines such as smallpox is subject to the usual ethical rules such as the need for informed consent. However, the participants will not often be at risk of catching the disease but expose themselves by taking part in the experimentation. Professor Deutsch explores the implications of this, including the position of vulnerable groups such as children, those with mental handicaps, and those acting under orders such as the miliary, the policy and fire officers. PMID- 16211736 TI - A randomized, placebo-controlled study of rofecoxib with paracetamol in early post-tonsillectomy pain in adults. AB - BACKGROUND AND OBJECTIVE: Effective and early treatment of postoperative pain and nausea have become pivotal for the early discharge of patients after tonsillectomy. Opioid-based analgesia is standard practice but the use of non steroidal anti-inflammatory drugs is discouraged due to their platelet inhibiting properties. The cyclo-oxygenase-2 selective non-steroidal anti-inflammatory drugs are effective analgesics and do not affect platelet function. We hypothesized that premedication with cyclo-oxygenase-2 selective non-steroidal anti inflammatory drug in addition to paracetamol would provide effective analgesia and decrease opioid consumption during early recovery from tonsillectomy. METHODS: In a randomized, placebo-controlled study of adult tonsillectomy patients (n=40) one group (R-group; n=20) was premedicated with paracetamol 1.5 g and rofecoxib 50 mg and a control group (P-group; n=20) was premedicated with paracetamol 1.5 g and placebo. Morphine was used as rescue medication. Postoperative (24 h) pain scores (0--10), morphine consumption as well as intraoperative blood loss were recorded. RESULTS: We found no overall difference in pain scores between the groups but significantly more patients in the placebo group had pain scores >5 within the first 8 h. The rofecoxib group consumed less morphine during the first 12 h. A lower intraoperative blood loss was observed in the rofecoxib group. CONCLUSION: Our results suggest an early although clinically minor analgesic benefit of the addition of a cyclo-oxygenase-2 selective non steroidal anti-inflammatory drug to paracetamol as premedication for adult tonsillectomy. PMID- 16211735 TI - Comparision of etomidate and propofol for fibreoptic intubation as part of an airway management algorithm:a prospective, randomizes, double-blind study. AB - BACKGROUND AND OBJECTIVE: In our algorithm for management of the anticipated difficult airway the induction agent (etomidate) is administered after the tip of the fibreoptic is placed in the trachea but before the tube is advanced over it. In a previous investigation we demonstrated the safety of this method. Due to its popularity as an induction agent, some would like to replace etomidate with propofol. However, because rapid recovery of spontaneous breathing is crucial with this technique, substitution might not be advisable. We compared the speed of recovery of spontaneous breathing after fibreoptic intubation between etomidate and propofol. METHODS: In this prospective, randomized, double-blind study we used either 0.2 mg kg[-1] etomidate or 2 mg kg[-1] propofol for induction. Our technique of nasotracheal fibreoptic intubation consists of using fentanyl, cocaine instillation into the lower nasal canals, cricothyroid injection of lidocaine, performing bronchoscopy, administration of etomidate and advancing the tube after loss of consciousness. We measured time to loss of consciousness, time to recovery of spontaneous breathing, lowest bi-spectral index value and time to lowest value. RESULTS: Time to loss of consciousness did not differ. The time to recovery of spontaneous breathing differed significantly: the median time (interquartile range [range]) for etomidate was 81 s (62--102 [0- 166]), and for propofol 146 s (95--260 [65--315]); P=0.001. The lowest bi spectral index values were not different. The time of the lowest bi-spectral index values differed significantly: for etomidate 58 s (51--68 [38--100]), and for propofol 90 s (52--125 [38--172]); P=0.015. CONCLUSION: For nasotracheal fibreoptic intubation, where the tube is advanced after induction of anaesthesia, we still recommend etomidate because spontaneous breathing recovers faster than with propofol. PMID- 16211737 TI - Politics, prohibitions and the lost public perspective: a comment on Bill C-56: the Assisted Human Reproduction Act. PMID- 16211738 TI - Clones, controversy, confusion and criminal law: a reply to Professor Caulfield. PMID- 16211739 TI - A randomized, double-blind, close-ranging, pilot study of intravenous granisetron in the prevention of postoperative nausea and vomiting in patients abdominal hysterectomy. AB - BACKGROUND AND OBJECTIVE: Postoperative nausea and vomiting (PONV) is a frequent and unpleasant experience that may increase postoperative complications and costs. For surgical procedures with a high risk of PONV, prevention is preferable to treatment. In this study, the authors explore the dose-response relationship between granisetron administered just prior to the end of surgery and post operative nausea and vomiting in patients undergoing abdominal hysterectomy. METHODS: This was a randomized, double-blind, placebo-controlled, pilot study of post-operative nausea and vomiting prevention. Patients undergoing elective open abdominal hysterectomy requiring general anaesthesia received a single dose of granisetron 0.1, 0.2 or 0.3 mg or placebo administered approximately 15 min prior to the end of surgery. The primary efficacy end-point was the proportion of patients with no vomiting in the 0--6 h interval following medication administration. No inferential statistics were planned. RESULTS: The proportion of patients with no vomiting episode in the 0--6 h interval after administration of study medication was higher in each granisetron treatment group (>90%) than in the placebo group (77%). Proportions of patients with no vomiting episodes in the 0--24 h interval were similar across treatment groups. Results of analyses of proportions of patients with no moderate or severe nausea episodes, proportions of those requiring rescue medication and times to first use of rescue medication suggested a treatment effect of granisetron relative to placebo in both the 0--6 and 0--24 h intervals. Similar proportions of patients in each treatment group reported at least one adverse event. CONCLUSIONS: Granisetron at doses of 0.1, 0.2 and 0.3 mg administered just prior to the end of surgery suggested a trend of improved efficacy compared to placebo in preventing postoperative nausea and vomiting in the first 6 h after abdominal hysterectomy. This pilot study did not identify a dose-response relationship. PMID- 16211740 TI - [The role of Waclawa Moszynski in the development of the Polish School of Machine and Mechanism Theory]. AB - The end of the 19th and the first half of the 20th centuries saw the emergence and intensive development of many disciplines in the technical sciences, and the laying of a groundwork for those disciplines in their current form. In Poland, a country deprived of independent statehood until 1918, this was additionally a period when native research centres and scientific schools came into being, and when Polish-language specialist literatures emerged as well. Many of the eminent personages whose activities contributed to those trends have already been described in biographical notes and articles. There are still, however, not enough studies dealing with the substance of their scientific and professional achievements. One of the personages in question was Waclawa Moszynski, professor of the Warsaw Technical University, a pioneer of machine construction and mechanism theory, author of the first academic textbook in the field to be published in Poland. The current article discusses Moszynski's contribution to the development of mechanism and machine theory. The first part of the article gives an outline of the history of the discipline until 1945, presents the context of the after-war activities of the author, and evaluates his influence on the development of machine and mechanism theory in Poland; it also carries a short biography of Moszynski. The rest of the article is devoted to Moszynski's scientific achievements, and describes his approach to matters of structure, kinematics and dynamics, with special focus on those of his formulations and solution to problems that appear particularly innovative and original. The article also points out those proposals by Moszynski which made him a precursor of other disciplines, such as vibroacoustics, biomechanics and ergonomics. The paper also presents the role of Moszynski's work as a foundation for the development of the Warsaw research-and-teching centre in the field. The achievements and methods of the Warsaw school combined with the those of other centres in Poland, which based on the earlier Lwow tradition, thus leading to very significant research and educational accomplishments. PMID- 16211741 TI - Cold shoulder treatment: the disposition of frozen embryos post-divorce. PMID- 16211742 TI - Eugenics and compulsory sterilization laws: providing redress for victims of a shameful era in United States history. PMID- 16211743 TI - A patient's right of privacy in computerized pharmacy records. PMID- 16211744 TI - Which is responsible for the haemodynamic response due to laryngoscopy and endotracheal intubation? Catecholamines, vasopressin or angiotensin? AB - BACKGROUND AND OBJECTIVE: We have investigated the concentrations of epinephrine, norepinephrine, vasopressin and angiotensin converting enzyme activity to explore the role of these mediators in the neuroendocrine response to laryngoscopy and tracheal intubation. METHODS: One hundred (50 male, 50 female) ASA I patients aged 20-50 yr (mean+/-SEM; 35.59+/-0.99) were included in the study. They were undergoing elective surgery under standard anaesthesia induction and maintenance using tracheal intubation. Plasma concentrations of epinephrine, norepinephrine and vasopressin as well as plasma angiotensin converting enzyme activity were determined at four time points, before (T1) and after (T2) induction, and 2 (T3) and 5 min (T4) after intubation. Blood pressure and heart rate were recorded at corresponding times to reveal if any correlation existed between haemodynamic parameters and neuroendocrine response. RESULTS: Heart rate increased after induction and intubation (P<0.05) and decreased significantly at T4 (P<0.05). Systolic blood pressure decreased significantly (P<0.05) after induction and increased slightly after intubation decreasing to below baseline value (P<0.05) at T4. Diastolic blood pressure increased slightly after intubation and decreased significantly (P<0.05) at T4. Plasma epinephrine and norepinephrine concentrations decreased after induction and increased at T3 and T4 without reaching significance. Vasopressin concentrations increased slightly at T2 and T3 and decreased significantly at T4 (P<0.05). Angiotensin converting enzyme activity was unaffected when compared with baseline values. CONCLUSIONS: Blood pressure, heart rate, plasma epinephrine, norepinephrine and vasopressin concentrations increased slightly in response to laryngoscopy and intubation, all returning to or below baseline 5 min later with no change in angiotensin converting enzyme activity in normotensive patients. PMID- 16211745 TI - The quiet battle for the heart of liberty--a victory for the cautious: Washington v. Glucksberg, 117 S. Ct. 2258 (1997). PMID- 16211746 TI - A maternal duty to protect fetal health? PMID- 16211747 TI - Medicare and Medicaid programs; condition of participation: immunization standard for long term care facilities. Final rule. AB - The goal of this final rule is to increase immunization rates in Medicare and Medicaid participating long term care (LTC) facilities by requiring LTC facilities to offer each resident immunization against influenza annually, as well as lifetime immunization against pneumococcal disease. LTC facilities will be required to ensure that before offering the immunization, each resident or the resident's legal representative receives education regarding the benefits and potential side effects of immunization. The facilities will be required to offer immunization against influenza annually and immunization against pneumococcal disease once, unless medically contraindicated or the resident or the resident's legal representative refuses immunization. Increasing the use of Medicare-funded preventive services is a goal of both CMS and the Centers for Disease Control and Prevention (CDC). This final rule is intended to increase the number of elderly receiving influenza and pneumococcal immunization and decrease the morbidity and mortality rate from influenza and pneumococcal diseases. PMID- 16211748 TI - Beyond Jaffee v. Redmond: should the federal courts recognize a right to physician-patient confidentiality? PMID- 16211749 TI - Racial disparities in the delivery of health care. PMID- 16211750 TI - Asymptomatic HIV and the Americans with Disabilities Act: Runnebaum v. Nationsbank of Maryland, N.A. PMID- 16211751 TI - Time and tidal volume-dependent ventilator-induced lung injury in healthy rats. AB - BACKGROUND AND OBJECTIVE: We evaluated the effect of duration of mechanical ventilation with different tidal volumes (VT) on ventilator-induced lung injury in healthy rats. METHODS: Anaesthetized rats were ventilated with VT between 9 and 45 mL kg[-1] for 1 or 7 h with a positive end-expiratory pressure of 2.5 cmH2O. RESULTS: After 1 h, rats ventilated even with the highest applied VT (36 and 45 mL kg[minus sign]1, resulting in average peak airway pressures of 30 +/-3 and 37 +/- 4 cmH2O), had no detectable alterations in dynamic or static lung mechanics, gas exchange or pulmonary permeability, but a moderate degree of lung inflammation (neutrophil accumulation in broncho-alveolar lavage) observed in all groups. In contrast, after 3 h of ventilation, rats ventilated with the highest VT (36 and 45 mL kg[minus sign]1) died from progressive circulatory failure and high-permeability pulmonary oedema, manifested by hypoxaemia, an increased alveolar-arterial protein concentration ratio and a reduced static lung compliance (mortality rate at 7 h, 62.5% and 100%). Animals with lower VT all survived and presented no changes in the measured variables. CONCLUSION: These results in normal rats demonstrate the preponderant effect of the duration (>3 h) of 'aggressive' ventilation and the cut-off value of the level of VT applied (>27 mL kg[minus sign]1). PMID- 16211752 TI - Marching to the beat of a different drummer: is military law and mental health out-of-step after Jaffee v. Redmond? PMID- 16211753 TI - Effect of ciprofloxin on the pharmacokinetics of intravenous lidocaine. AB - BACKGROUND AND OBJECTIVE: Recent studies have suggested that cytochrome P-450 isoenzyme 1A2 has an important role in lidocaine biotransformation. We have studied the effect of a cytochrome P-450 1A2 inhibitor, ciprofloxacin, on the pharmacokinetics of lidocaine. METHODS: In a randomized, double-blinded, cross over study, nine healthy volunteers ingested for 2.5 days 500 mg oral ciprofloxacin or placebo twice daily. On day 3, they received a single dose of 1.5 mg kg[-1] lidocaine intravenously over 60 min. Plasma concentrations of lidocaine, 3-hydroxylidocaine and monoethylglycinexylidide were determined for 11 h after the start of the lidocaine infusion. RESULTS: Ciprofloxacin increased the mean peak concentration and area under plasma concentration-time curve of lidocaine by 12% (range [-6] to+46%; P<0.05) and 26% (8--59%; P 0.01), respectively. The mean plasma clearance of lidocaine was decreased by ciprofloxacin by 22% (7--38%; P<0.01). Ciprofloxacin decreased the area under the plasma concentration-time curve of monoethylglycinexylidide by 21% (P<0.01) and that of 3-hydroxylidocaine by 14% (P< 0.01). CONCLUSION: The plasma decay of intravenously administered lidocaine is modestly delayed by concomitantly administered ciprofloxacin. Ciprofloxacin may increase the systemic toxicity of lidocaine. PMID- 16211754 TI - Bridging the gap between life insurer and consumer in the genetic testing era: the RF proposal. PMID- 16211755 TI - The international human right to health: what does this mean for our nation and world? PMID- 16211756 TI - Forcible medication and personal autonomy: the case of Charles Thomas Sell. PMID- 16211757 TI - The development of the undue burden standard in Stenberg v. Carhart: will proposed RU-486 legislation survive? PMID- 16211758 TI - Neuraxial blockade and patient risk. PMID- 16211759 TI - Informed consent and patients' rights in Japan. PMID- 16211760 TI - Prenatal tort law and the personhood of the unborn child: a separate legal existence. PMID- 16211761 TI - Race as a proxy for drug response: the dangers and challenges of ethnic drugs. PMID- 16211762 TI - What if technology never stops improving? Medicare's future under continuous cost increases. PMID- 16211763 TI - Lack of insurance coverage for prescription contraception by an otherwise comprehensive plan as a violation of Title VII as amended by the Pregnancy Discrimination Act--stretching the statute too far. PMID- 16211764 TI - Medically necessary organ transplants for prisoners: who is responsible for payment? PMID- 16211765 TI - Gene patents and the death of dualism. PMID- 16211766 TI - Squandering the gain: gainsharing and the continuing dilemma of physician financial incentives. PMID- 16211767 TI - Public opinion and policymaking in the states: the case of post-Roe abortion policy. AB - Whether public opinion should be expected to play a role in the shaping of abortion legislation in the states is a debatable question. Representation is a difficult task, especially if legislators receive mixed cues from the public, activists, and the political parties. In this study, we find that grass-roots activism and public opinion tend to match, and both are reflected in state abortion policy. In addition, more Pro-Life policies are found in states with a tradition of conservative policies in other areas, Republican majorities in the state legislature, more Catholic residents, and fewer women legislators. These patterns hold true for a composite index of abortion policies and for the specific policy area of government funding of abortions. Slightly different patterns occur for parental consent law, though these statutes also tend to reflect general preferences on abortion and interest group activity in a state. PMID- 16211768 TI - The rising agenda of physician-assisted suicide: explaining the growth and content of morality policy. AB - Employing theories and methods of agenda-setting analysis, this article explains the rapid rise of physician-assisted suicide (PAS) on the national political agenda based on its status as a morality policy. PAS reached the mass agenda before the professional agenda, probably because PAS is an outgrowth of previous right-to-die policies and Dr. Jack Kevorkian's assisted suicides provided major focusing events. As in other morality policies, competing groups fight for the last word, but PAS has been blocked on most governmental agendas because its image and media tone has been mostly negative and public opinion is divided. Groups in a few generally liberal states have tried to enact policy through referenda when legislators failed to address the issue. We speculate that competing interest groups will become more active and that state courts will become a venue of choice in the future. PMID- 16211769 TI - [The evaluation of arterial hypertension in a patient with arteritis is central to the assessment of renal artery stenosis]. PMID- 16211770 TI - [Epidemiology of arteriopathy of the lower limbs: a frequent but underestimated disease]. PMID- 16211771 TI - [Management of obliterating arteriopathy of the lower limbs in the diabetic patient]. PMID- 16211772 TI - Resource guide for use with Hispanic nephrology patients. PMID- 16211773 TI - [Cellular therapy in atherosclerotic arteriopathies: is there hope for those with severe prognosis?]. PMID- 16211774 TI - ANNA Position Statements (revised or reaffirmed April 2005). PMID- 16211775 TI - Limited benefit of combined use of tar-based shampoo with 50 microg/ml calcipotriol solution in scalp psoriasis. PMID- 16211776 TI - Jesuits in Spanish America: contributions to the exploration of the American materia medica. PMID- 16211777 TI - Concordant occipital encephalocele in monoamniotic twins. AB - Anomalies occur with a greater frequency in twin gestations. Due to its multifactorial inheritance, twins are usually discordant for encephalocele. We present a case of monoamniotic twins concordant for occipital encephalocele and discordant for lung and cord anomalies. Ultrasonographic examination at 17 weeks' gestation revealed occipital encephalocele in both fetuses. The maternal serum level of alpha-fetoprotein was increased. Fetal autopsy revealed occipital encepaholocele in both twins and right pulmonary hypoplasia and one umbilical artery in one sibling. Monoamniotic twins concordant for encephalocele occur with extreme rarity. To the best of our knowledge, monoamniotic twins concordant for this neural tube defect have not been previously reported. PMID- 16211778 TI - Changes of lactate, glucose, ionized calcium and glutamate concentrations in cephalic vein blood during brain hypothermia using extracorporeal membrane oxygenation (ECMO) in a newborn infant with hypoxic-ischemic encephalopathy. AB - An asphyxiated infant with severe hypoxic-ischemic encephalopathy was treated by brain hypothermia using extracorporeal membrane oxygenation (ECMO). The brain hypothermia using ECMO maintained cardiopulmonary functions and stabilized brain temperatures by stably supply of the cooled blood to the brain. Moreover,we measured the levels of glucose, lactate, ionized calcium and glutamate in plasma. The comparison between these levels in the artery and in the cephalic vein, suggests that glucose, lactate, ionized calcium and glutamate might be used as markers of the effects of hypothermia therapy. PMID- 16211779 TI - Joint SNM/RSNA Molecular Imaging Summit Statement. PMID- 16211780 TI - Treatment of symptomatic congenital cytomegalovirus infection with valganciclovir. AB - Cytomegalovirus (CMV) is the most common cause of congenital infection in humans. Some congenitally infected infants will develop sequelae later in life, especially sensorineural hearing loss (SNHL) and mental retardation. There is no generally accepted antiviral therapy for the treatment of symptomatic congenital CMV infections yet. We present a neonate with symptomatic congenital CMV infection, who was treated with intravenous (iv) ganciclovir (GCV) during 18 days and subsequently with oral valganciclovir (VGCV) for 5.5 months, in an attempt to prevent development of SNHL. GCV was given intravenously 10 mg/kg/day in two doses and VGCV doses ranged from 280-850 mg/m2 bidaily (bid). Our experience shows that it is not possible to give a fixed dosing regime for VGCV in neonates and that continuous adaptation of dose is necessary to achieve stable target levels of GCV and to keep the viral load in urine at undetectable level. At 18 months of age no hearing deterioration has occurred. While the current findings are encouraging, the limitations of a single case report with a relatively short follow-up emphasizes the need for further prospective randomized studies to evaluate pharmacokinetics, efficacy and safety of VGCV therapy in neonates with congenital CMV infection. PMID- 16211781 TI - Ehrlich's theory of drug action and anti-tuberculous gold therapy. PMID- 16211782 TI - Rational souls and the beginning of life (a reply to Robert Pasnau). PMID- 16211783 TI - Wrongful life. PMID- 16211784 TI - Neuraxial blockade and patient risk. PMID- 16211785 TI - Ethical issues in biological engineering. PMID- 16211786 TI - A conceptual framework for genetic policy: comparing the medical, public health, and fundamental rights models. PMID- 16211787 TI - General anaesthesia for a super obese patient. PMID- 16211788 TI - Intraperitoneal tramadol and buvacaine in total abdominal hysterectomy. PMID- 16211789 TI - Recombinant activated factor vii to control massive post operative bleeding after septic aortobifemoral grafting. PMID- 16211790 TI - Sub-anaesthestic doses of ketamine impair cardiac parasympathetic regulation. PMID- 16211792 TI - More California teens consume soda and fast food each day than five servings of fruits and vegetables. PMID- 16211791 TI - Intraoperative metamizol as cause for acute anaphylactic collapse. PMID- 16211793 TI - How primary care practice affects Medicaid patients' use of emergency services. AB - The use of emergency departments (EDs) in the U.S. continues to rise. Some of these ED visits may reflect limited access to primary care, even among patients with a primary care provider. Payers and policymakers have tried to restrain ED use, because of concern over high charges and discontinuity of care. But most of these attempts have involved erecting financial and administrative barriers to going to the ED, rather than expanding access to primary care. Is it possible to reduce excessive use of EDs by making primary care practices more "user friendly"? This Issue Brief summarizes research that identifies primary care characteristics associated with ED use in a Medicaid managed care population. It suggests a strategy to simultaneously improve access to primary care and reduce costs of ED care. PMID- 16211794 TI - No time to spare: improving access to trauma care. AB - Since the September 11 attacks, policymakers are paying increasing attention to the adequacy of the U.S. trauma care system to handle potential mass casualty incidents. This attention has led to questions about how well the trauma system covers the population for day-to-day trauma, such as motor vehicle accidents and gunshot wounds. Although the number of trauma centers has increased in the last decade, no national plan exists to assure that everyone has timely access to a specialized trauma center if needed. This Issue Brief summarizes a new study that estimates the proportion of residents that can reach a trauma center by ground or air ambulance within one hour of where they live, using objective measures of travel times and distances. PMID- 16211796 TI - [Remarks concerning the article "Physiology, role and neuropharmacology of yawning" by U.R. Goessler et al. Laryngo-Rhino-Otol 2005;84:345-351]. PMID- 16211797 TI - Weighted Zernike expansion with applications to the optical aberration of the human eye. AB - We propose to use weighted Zernike functions to represent the aberrations of an eye. Methods for computing the phase of an aberrated ophthalmic wavefront in terms of weighted Zernike functions are discussed. In particular, we consider several options for integrating the phase out of its measured slopes. The weighted functions involve a free parameter. Clinical data on subjective refraction and aberration maps of individual subjects are used to determine an estimate for this parameter. PMID- 16211799 TI - Study on the effects of monochromatic aberrations in the accommodation response by using adaptive optics. AB - The effect of asymmetric monochromatic aberrations in the accommodation response was studied by using an adaptive optics (AO) system. This approach permits the precise modification of ocular aberrations during accommodation. The AO system is composed of a real-time Hartmann-Shack wavefront sensor and a membrane deformable mirror with 37 independent actuators. The accommodation response was measured in two subjects with their normal aberrations and with the asymmetric aberrations terms corrected. We found a significant and systematic increase in the response accommodation time, and a reduction in the peak velocity, in both subjects when the aberrations were corrected in real time. However, neither the latency time nor the precision of the accommodation were affected. These results may indicate that the monochromatic aberrations play a role in driving the accommodation response. PMID- 16211798 TI - Spectrum analysis of motion parallax in a 3D cluttered scene and application to egomotion. AB - Previous methods for estimating observer motion in a rigid 3D scene assume that image velocities can be measured at isolated points. When the observer is moving through a cluttered 3D scene such as a forest, however, pointwise measurements of image velocity are more challenging to obtain because multiple depths, and hence multiple velocities, are present in most local image regions. We introduce a method for estimating egomotion that avoids pointwise image velocity estimation as a first step. In its place, the direction of motion parallax in local image regions is estimated, using a spectrum-based method, and these directions are then combined to directly estimate 3D observer motion. There are two advantages to this approach. First, the method can be applied to a wide range of 3D cluttered scenes, including those for which pointwise image velocities cannot be measured because only normal velocity information is available. Second, the egomotion estimates can be used as a posterior constraint on estimating pointwise image velocities, since known egomotion parameters constrain the candidate image velocities at each point to a one-dimensional rather than a two-dimensional space. PMID- 16211800 TI - Depth resolution and displayable depth of a scene in three-dimensional images. AB - The depth resolution and the recordable object depth range, obtainable with parallel, toed-in and sliding aperture camera configurations for multiview image acquisition in the three-dimensional imaging systems, are found by assuming that the camera lens resolution is diffraction limited and the resolution of the recorded image is limited by a pixel pitch of the imaging sensor. The depth resolution for the holographic image is calculated and compared with that of the multiview images for the same parameter values. The influence of the viewer's eye resolution limit on the depth resolution of the multiview images and hologram is also found. PMID- 16211802 TI - Fuzzy-rule-based image reconstruction for positron emission tomography. AB - Positron emission tomography (PET) and single-photon emission computed tomography have revolutionized the field of medicine and biology. Penalized iterative algorithms based on maximum a posteriori (MAP) estimation eliminate noisy artifacts by utilizing available prior information in the reconstruction process but often result in a blurring effect. MAP-based algorithms fail to determine the density class in the reconstructed image and hence penalize the pixels irrespective of the density class. Reconstruction with better edge information is often difficult because prior knowledge is not taken into account. The recently introduced median-root-prior (MRP)-based algorithm preserves the edges, but a steplike streaking effect is observed in the reconstructed image, which is undesirable. A fuzzy approach is proposed for modeling the nature of interpixel interaction in order to build an artifact-free edge-preserving reconstruction. The proposed algorithm consists of two elementary steps: (1) edge detection, in which fuzzy-rule-based derivatives are used for the detection of edges in the nearest neighborhood window (which is equivalent to recognizing nearby density classes), and (2) fuzzy smoothing, in which penalization is performed only for those pixels for which no edge is detected in the nearest neighborhood. Both of these operations are carried out iteratively until the image converges. Analysis shows that the proposed fuzzy-rule-based reconstruction algorithm is capable of producing qualitatively better reconstructed images than those reconstructed by MAP and MR P algorithms. The reconstructed images a resharper, with small features being better resolved owing to the nature of the fuzzy potential function. PMID- 16211801 TI - Optimized stereo reconstruction of free-form space curves based on a nonuniform rational B-spline model. AB - Analytical reconstruction of 3D curves from their stereo images is an important issue in computer vision. We present an optimization framework for such a problem based on a nonuniform rational B-spline (NURBS) curve model that converts reconstruction of a 3D curve into reconstruction of control points and weights of a NURBS representation of the curve, accordingly bypassing the error-prone point to-point correspondence matching. Perspective invariance of NURBS curves and constraints deduced on stereo NURBS curves are employed to formulate the 3D curve reconstruction problem into a constrained nonlinear optimization. A parallel rectification technique is then adopted to simplify the constraints, and the Levenberg-Marquardt algorithm is applied to search for the optimal solution of the simplified problem. The results from our experiments show that the proposed framework works stably in the presence of different data samplings, randomly posed noise, and partial loss of data and is potentially suitable for real scenes. PMID- 16211803 TI - Simultaneous spatial and spectral imaging of lasing droplets. AB - We present an experimental technique that allows the simultaneous spatial imaging and spectral analysis of falling droplets that exhibit lasing. Single droplet investigations serve as, among other purposes, a preliminary study for spray and combustion researchers. The described setup provides a valuable tool for the evaluation of microdroplet investigations with laser-spectroscopic techniques that rely on laser-induced fluorescence (LIF) or similar spectroscopical phenomena. The emphasis is that both spatial and spectral information are obtained from single-shot images of a falling droplet. Furthermore, combining spatial imaging and a spatially resolving optical multichannel analyzer makes a pointwise rastering of the droplets spectrum possible. This allows for the (almost) unambiguous determination of sources of influence on the spectrum of these droplets-such as geometrical distortion and lasing, nondissolved tracer lumps, and similar phenomena. Although the focus is on the experimental technique itself, we supplement detailed studies of lasing in falling microdroplets. These results were obtained with the aim of developing a system for measuring temperature distributions in droplets and sprays. In the light of these results the practice of calibrating a droplets spectrum by use of a bulk liquid sample needs to be critically reviewed. PMID- 16211804 TI - Imaging systems based on the encoding of optical coherence functions. AB - An imaging scheme is described that is based on the transmission of image-forming information encoded within optical coherence functions. The scheme makes use of dynamic random-valued encoding-decoding masks placed in the input-output planes of any linear optical system. The mask transmittance functions are complex conjugates of each other, as opposed to a similar coherence encoding scheme proposed earlier by two of this paper's authors that used identical masks. [Rhodes and Welch, in Euro-American Workshop on Optoelectronic Information Processing, SPIE Critical Review Series (SPIE, 1999), Vol. CR74, p. 1]. General analyses of the two coherence encoding schemes are performed by using the more general mutual coherence function as opposed to the mutual intensity function used in the earlier scheme. The capabilities and limitations of both encoding schemes are discussed by using simple examples that combine the encoding-decoding masks with free-space propagation, passage through a four-f system, and a single lens imaging system. PMID- 16211805 TI - Effects of occlusion, edges, and scaling on the power spectra of natural images. AB - The circularly averaged power spectra of natural image ensembles tend to have a power-law dependence on spatial frequency with an exponent of approximately -2. This phenomenon has been attributed to object occlusion, the presence of edges, and scaling of object sizes (self-similarity) in natural scenes, although the relative importance of these properties is still unclear. A detailed examination of the effects of occlusion, edges, and self-similarity on the behavior of the power spectrum is conducted using a simple model of natural images. Numerical simulations show that edges and self-similarity are necessary for a power-law power spectrum over a wide range of spatial frequencies. Object occlusion is not an essential factor. A theoretical analysis for images containing nonoccluding objects supports these results. PMID- 16211806 TI - Coincidence fractional Fourier transform implemented with partially coherent light radiation. AB - We introduce the coincidence fractional Fourier transform (FRT) implemented with incoherent and partially coherent light radiation. Optical systems for implementing the coincidence FRT are designed. The results show that the visibility and quality of the coincidence FRT of an object are closely related to the light source's transverse size, coherence, and spectral width. As an example, we numerically study the coincidence FRT of a single slit. PMID- 16211807 TI - Numerical study on spectral properties of tungsten one-dimensional surface-relief gratings for spectrally selective devices. AB - Spectral properties of one-dimensional tungsten gratings with various depths and widths of grooves were investigated by means of finite-difference time-domain simulation with a multi-Lorentz model. Shallow gratings showed a strong absorption peak due to surface plasmon polaritons when the oscillation of the electric field was parallel to the grating vector. On the other hand, deep gratings with wide grooves showed a different absorption attributed to the microcavity effect when the oscillation of the electric field was perpendicular to the grating vector. With narrowed grooves, another absorption with d dependence occurred, which was probably due to vertically oscillating surface plasmons to the grooves. PMID- 16211808 TI - Axial irradiance of a focused beam. AB - The principal maximum of axial irradiance of a focused beam with a low Fresnel number does not lie at its focal point; instead it lies at a point that is closer to the focusing pupil. It has been shown by the numerical example of a weakly truncated Gaussian beam that its value increases and its location moves closer to the pupil when spherical aberration is introduced into the beam. Such an increase has been referred to as "beyond the conventional diffraction limit." Similarly, an increase in the value and a shift in the location of the principal maximum of axial irradiance of a uniform beam toward the pupil by the introduction of some spherical aberration has been characterized as an unexpected result. We explain why and how such a result comes about and that it neither invalidates any diffraction limit nor is it unexpected. We illustrate this for uniform as well as Gaussian beams of various truncation ratios. Both focused and collimated beams aberrated by spherical aberration or astigmatism are considered. PMID- 16211809 TI - Strehl ratio of a Gaussian beam. AB - We discuss the Strehl ratio of systems with a Gaussian pupil and determine the range of validity of its approximate expression based on the aberration variance. The results given are equally applicable to propagation of Gaussian beams. The uniform and weakly truncated pupils are considered as limiting cases of a Gaussian pupil. We show that the approximate expression for Strehl ratio in terms of the aberration variance yields a good estimate of the true value for a strongly truncated pupil but a much smaller value for a weakly truncated pupil. PMID- 16211810 TI - Diffraction characteristics with various polarizations of overlapping holographic gratings in a uniaxial crystal. AB - By the Riemann method, a coupled wave model is derived for the ordinary-to ordinary (OO) and extraordinary-to-extraordinary (EE) Bragg diffraction of a Gaussian beam by overlapping holographic gratings in a uniaxial crystal. The computer simulation is used to discuss the relations among the diffraction efficiency, the index modulation, the wavelength sensitivity, the angular sensitivity, and the the widths of the recording and reading beams. The characteristics of EE and OO diffraction in a uniaxial crystal are found to be remarkably different. The simulation shows that EE diffraction may exhibit far higher diffraction efficiency than does OO diffraction for very low index modulation with the same hologram size, for example, nearly 90% when the size is 8.2 x 10(-5). PMID- 16211811 TI - Perfectly matched layers as nonlinear coordinate transforms: a generalized formalization. AB - A modified formulation of Maxwell's equations is presented that includes a complex and nonlinear coordinate transform along one or two Cartesian coordinates. The added degrees of freedom in the modified Maxwell's equations allow one to map an infinite space to a finite space and to specify graded perfectly matched absorbing boundaries that allow the outgoing wave condition to be satisfied. The approach is validated by numerical results obtained by using Fourier-modal methods and shows enhanced convergence rate and accuracy. PMID- 16211812 TI - Electromagnetic computational method for the scattering of an axisymmetric laser beam by an inhomogeneous body of revolution. AB - An original computational method for solving the two-dimensional problem of the scattering of an axisymmetric laser beam by an arbitrary-shaped inhomogeneous body of revolution is presented. This method relies on a domain decomposition of the scattering zone into concentric spherical radially homogeneous subdomains and on an expansion of the angular dependence of the fields on the Legendre functions. Numerical results for the fields obtained for various scatterer geometries are presented and analyzed. PMID- 16211813 TI - Revised Kubelka-Munk theory. III. A general theory of light propagation in scattering and absorptive media. AB - A generally applicable theoretical model describing light propagating through turbid media is proposed. The theory is a generalization of the revised Kubelka Munk theory, extending its applicability to accommodate a wider range of absorption influences. A general expression for a factor taking into account the effect of scattering on the total photon path traversed in a turbid medium is derived. The extended model is applied to systems of ink-dyed paper sheets mixtures of wood fibers with dyes-which represent examples of systems that have thus far eluded the original Kubelka-Munk model. The results of simulations of the spectral dependence of Kubelka-Munk coefficients of absorption and scattering show that they compare very well with those derived from experimental results. PMID- 16211814 TI - Direct characterization and removal of interfering absorption trends in two-layer turbid media. AB - We propose a method to isolate absorption trends confined to the lower layer of a two-layer turbid medium, as is desired in near-infrared spectroscopy (NIRS) of cerebral hemodynamics. Several two-layer Monte Carlo simulations of NIRS time series were generated using a physiologically relevant range of optical properties and varying the absorption coefficients due to bottom-layer, top layer, and/or global fluctuations. Initial results showed that by measuring absorption trends at two source-detector separations and performing a least squares fit of one to the other, processed signals strongly resemble the simulated bottom-layer absorption properties. Through this approach, it was demonstrated that fitting coefficients can be estimated within less than +/- 2% of the ideal value without any a priori knowledge of the optical properties present in the model. An analytical approximation for the least-squares coefficient provides physical insight into the nature of errors and suggests ways to reduce them. PMID- 16211815 TI - Light diffusion through a turbid parallelepiped. AB - Solutions of the diffusion approximation to the radiative transport equation are derived for a turbid (rectangular) parallelepiped using the method of image sources and applying extrapolated boundary conditions. The derived solutions are compared with Monte Carlo simulations in the steady-state and time domains. It is found that the diffusion theory is in good agreement with Monte Carlo simulations provided that the light is detected sufficiently far from the incident beam. Applications of the derived solutions, including the determination of the optical properties of the turbid parallelepiped, are discussed. PMID- 16211816 TI - Superresolution in total internal reflection tomography. AB - We simulate a total internal reflection tomography experiment in which an unknown object is illuminated by evanescent waves and the scattered field is detected along several directions. We propose a full-vectorial three-dimensional nonlinear inversion scheme to retrieve the map of the permittivity of the object from the scattered far-field data. We study the role of the solid angle of illumination, the incident polarization, and the position of the prism interface on the resolution of the images. We compare our algorithm with a linear inversion scheme based on the renormalized Born approximation and stress the importance of multiple scattering in this particular configuration. We analyze the sensitivity to noise and point out that using incident propagative waves together with evanescent waves improves the robustness of the reconstruction. PMID- 16211817 TI - Controllable dark-hollow beams and their propagation characteristics. AB - A new mathematical model called "controllable dark-hollow beams" is introduced to describe hollow beams. The central dark size of this beam can be controlled easily by the beam order N and parameter epsilon. An analytical formula is derived for the propagation of a controllable dark-hollow beam through a paraxial optical system, and some numerical calculations are carried out. Some important propagation characteristics of this beam, such as the beam propagation factor and the kurtosis parameter, are studied in detail, and their variation rules versus the beam order N and parameter epsilon are presented and plotted. PMID- 16211818 TI - Transverse intensity distributions of a broadband laser modulated by a hard-edged aperture. AB - By means of the Huygens-Fresnel diffraction integral, the field representation of a laser beam modulated by a hard-edged aperture is derived. The near-field and far-field transverse intensity distributions of the beams with different bandwidths are analyzed by using the representation. The numerical calculation results indicate that the amplitudes and numbers of the intensity spikes decrease with increasing bandwidth, and beam smoothing is achieved when the bandwidth takes a certain value in the near field. In the far field, the radius of the transverse intensity distribution decreases as the bandwidth increases, and the physical explanation of this fact is also given. PMID- 16211819 TI - Theory of the unstable Bessel resonator. AB - A rigorous analysis of the unstable Bessel resonator with convex output coupler is presented. The Huygens-Fresnel self-consistency equation is solved to extract the first eigenmodes and eigenvalues of the cavity, taking into account the finite apertures of the mirrors. Attention was directed to the dependence of the output transverse profiles; the losses; and the modal-frequency changes on the curvature of the output coupler, the cavity length, and the angle of the axicon. Our analysis revealed that while the stable Bessel resonator retains a Gaussian radial modulation on the Bessel rings, the unstable configuration exhibits a more uniform amplitude modulation that produces output profiles more similar to ideal Bessel beams. The unstable cavity also possesses higher-mode discrimination in favor of the fundamental mode than does the stable configuration. PMID- 16211820 TI - Rotational invariance approach for the evaluation of multiple phases in interferometry in the presence of nonsinusoidal waveforms and noise. AB - Incorporation of two phase-shifting devices in a holographic moire configuration not only renders the interferometer compatible with automated measurements but also allows for simultaneous measurement of multiple phase information in the interferometer. However, simultaneous handling of multiple phase steps and subsequent simultaneous determination of multiple phase distributions requires the introduction of novel tools in phase-shifting interferometry. In this context, the aim of this paper is to propose a subspace invariance approach to address these issues. This approach takes advantage of the rotational invariance of signal subspaces spanned by two temporally displaced data sets formed from the intensity fringes recorded temporally on pixels of the CCD camera. The method first identifies the arbitrary phase steps imparted to the piezoactuator devices. The estimated phase steps are subsequently applied in the linear Vandermonde system of equations to determine the phase distributions. The method also allows for handling nonsinusoidal wavefronts. Since the phase steps are extracted at every point on the interferogram, the method is applicable to configurations that use spherical beams. The robustness of the method is investigated by adding white Gaussian noise during the simulations. PMID- 16211821 TI - Iterative algorithm for subaperture stitching interferometry for general surfaces. AB - A novel iterative algorithm for subaperture stitching interferometry for general surfaces is presented. It is based on the alternating optimization technique and the successive linearization method. The computer-aided-design model of the tested surface is used to determine the overlapping region precisely. Subapertures are simultaneously stitched by minimizing deviations among them as well as deviations from the nominal surface. Precise prior knowledge of the six degrees-of-freedom nulling and alignment motion is no longer required. PMID- 16211822 TI - Quantum-statistical analysis of multimode far-infrared and submillimeter-wave astronomical interferometers. AB - We present a detailed quantum-statistical model of multimode far-infrared and submillimeter-wave astronomical interferometers. The scheme identifies explicitly the optical modes associated with each telescope and uses these to trace the quantum-statistical properties of the field from a source through the telescopes, through the beam combiners, and onto the detectors. The scheme can be used with any optical configuration, and elegant expressions result for the average rate at which photons are detected by the pixels of an imaging array, the mean-square fluctuations in the rates, and the correlations between the fluctuations in the rates of different pixels. Numerous extensions to the basic technique are possible. PMID- 16211823 TI - Fast analysis method for polarization-dependent performance of a concave diffraction grating with total-internal-reflection facets. AB - A fast simulation method for a waveguide-based concave grating with total internal-reflection (TIR) facets is presented using the Kirchhoff-Huygens principle. Unlike the conventional scalar method, modifications are made to take into account the influence of the Goos-Hanchen (GH) shift. The simple method is in good agreement with a numerical method based on rigorous coupled-wave analysis for a wide range of practical device parameters and can provide an insightful physical explanation for the numerical results. It is shown that the GH shift is a main contributing factor for the loss and the polarization-dependent loss of an etched diffraction grating demultiplexer with TIR facets. PMID- 16211824 TI - Extending the Clapper-Yule model to rough printing supports. AB - The Clapper-Yule model is the only classical spectral reflection model for halftone prints that takes explicitly into account both the multiple internal reflections between the print-air interface and the paper substrate and the lateral propagation of light within the paper bulk. However, the Clapper-Yule model assumes a planar interface and does not take into account the roughness of the print surface. In order to extend the Clapper-Yule model to rough printing supports (e.g., matte coated papers or calendered papers), we model the print surface as a set of randomly oriented microfacets. The influence of the shadowing effect is evaluated and incorporated into the model. By integrating over all incident angles and facet orientations, we are able to express the internal reflectance of the rough interface as a function of the rms facet slope. By considering also the rough interface transmittances both for the incident light and for the emerging light, we obtain a generalization of the Clapper-Yule model for rough interfaces. The comparison between the classical Clapper-Yule model and the model extended to rough surfaces shows that the influence of the surface roughness on the predicted reflectance factor is small. For high-quality papers such as coated and calendered papers, as well as for low-quality papers such as newsprint or copy papers, the influence of surface roughness is negligible, and the classical Clapper-Yule model can be used to predict the halftone-print reflectance factors. The influence of roughness becomes significant only for very rough and thick nondiffusing coatings. PMID- 16211825 TI - Quasi-analytic formalism for mode characteristics in highly overmoded rectangular dielectric waveguide bends. AB - A fast and simple quasi-analytic method to simulate mode characteristics in highly overmoded rectangular dielectric waveguide bends is presented. Fast mode based bend models are necessary, since overmoded rectangular waveguides have become very popular in optical interconnects on printed circuit boards. The proposed method combines a simple mode solver with the formalism that was proposed by Melloni et al. [J. Lightwave Technol. 16, 571 (2001)], yielding to a very convenient and accurate quasi-analytic formalism for the bend transfer function based on matrix notation. For that purpose, a simple method to approximate leaky modes is introduced. The model offers the ability to predict individual modal phases and amplitudes within a given bend as well as the calculation of coupling losses and was validated using three-dimensional beam propagation-method simulation software. PMID- 16211826 TI - Propagation of Hermite-Gaussian and Laguerre-Gaussian beams beyond the paraxial approximation. AB - On the basis of the vectorial Rayleigh-Sommerfeld formulas and by means of the relation between Hermite and Laguerre polynomials, the analytical expressions for the propagation of the Hermite-Gaussian (HG) and Laguerre-Gaussian (LG) beams beyond the paraxial approximation are derived, with the corresponding far-field propagation expressions and that for the Gaussian beams being given as special cases of the results. Some detailed comparisons of our results with the expansion series and paraxial expressions are made, which show the advantages of our results over the expansion series. With the results obtained, some typical intensity patterns of nonparaxial HG and LG beams are shown. PMID- 16211827 TI - Propagation of polarized light through two- and three-layer anisotropic stacks. AB - The extended Jones formulation is used to investigate propagation at nonnormal incidence through two- and three-layer systems of birefringent material in which the optic axes of the individual layers are in the plane of the layers. Such systems are equivalent to two optical elements in series-an equivalent retardation plate and a polarization rotator. Analytical solutions are obtained for the equivalent retardation and rotation. The major finding is that, in general, there are two nonnormal incidence directions for which the retardation vanishes; therefore these two directions are optic axes of the composite system. These simple layered systems therefore behave in a manner similar to biaxial crystals. Moreover, the results illustrate the fact that even if the optic axes of individual layers in composite systems are in the plane of the layers, the optic axes of the system are, in general, out of this plane. PMID- 16211828 TI - Variable aberration generators using rotated Zernike plates. AB - The rotational properties of Zernike polynomials allow for an easy generation of variable amounts of aberration using two rotated phase plates, each one encoding one or several Zernike modes. This effect may be used to build variable aberration generators useful for calibrating different kinds of aberrometer. PMID- 16211829 TI - [Tumor hypoxia:advantage or disadvantage?]. PMID- 16211830 TI - [The system the process or the patient?]. PMID- 16211831 TI - [Medical preparedness for emergencies]. PMID- 16211832 TI - [Tapeworm disease]. PMID- 16211833 TI - [Ankylosing spondylitis]. PMID- 16211835 TI - [Sudden vision impairement]. PMID- 16211834 TI - [Optical coherence tomography of macular layers]. PMID- 16211836 TI - [Skin necrosis of the uremic patient]. PMID- 16211837 TI - [Dairy products and acne]. PMID- 16211838 TI - [Care of sepsis in adults]. PMID- 16211839 TI - [ Lauri Ahlgren: Organism changes]. PMID- 16211841 TI - [Surgical antimicrobial prophylaxis:to whom, when and how?]. PMID- 16211840 TI - [One fifth of the hospital infections could be prevented]. PMID- 16211842 TI - [Long term care units--nest of resistant bacteria?]. PMID- 16211843 TI - [Use of antibiotics in hospital--to guide or to limit]. PMID- 16211844 TI - [Prevention of central vein catheter infections]. PMID- 16211845 TI - [Hand washing, cornerstone to prevent spreading of microbes]. PMID- 16211846 TI - [The bacterial cell cycle: DNA replication, nucleoid segregation, and cell division]. AB - Data on the bacterial cell cycle published in the last 10-15 years are considered, with a special stress on studies of nucleoid segregation between dividing cells. The degree of similarity between the eukaryotic mitotic apparatus and the apparatus performing nucleoid separation is discussed. PMID- 16211847 TI - [Regulation of metabolic and electron transport pathways in the freshwater bacterium Beggiatoa leptomitiformis D-402]. AB - The biomass yield of freshwater filamentous sulfur bacteria of the genus Beggiatoa, when grown lithoheterotrophically or mixotrophically, has been shown to increase 2 to 2.5 times under microaerobic conditions (0.12 mg/l oxygen) as compared to aerobic conditions (9 mg/l oxygen). The activity of the glyoxylate cycle key enzymes have been found to increase two to three times under microaerobic conditions (at an O2 concentration of 2 mg/l), and the activities of the sulfur metabolism enzymes increased three to five times (at an O2 concentration of 0.1-0.5 mg/l). It has also been found that, under microaerobic conditions, thiosulfate was almost completely oxidized to sulfate by the bacteria, without accumulation of intermediate metabolites. At the same time, a 2 to 15-fold decrease in the activities of the tricarboxylic acid cycle enzymes involved in the reduction of NAD and FAD was observed. Reorganization of the respiratory chain after changes in aeration and type of nutrition was also observed. It has been found that, in cells grown heterotrophically, the terminal part of the respiratory chain contained an aa3-type oxidase, whereas, during mixotrophic, lithoheterotrophic, and autotrophic growth, aa3-type oxidase synthesis was inhibited, and the synthesis of a cbb3-type oxidase, which is induced under microaerobic conditions, was activated. The gene of the catalytic subunit CcoN of the cbb3-type oxidase was sequenced and proved to be highly homologous to the corresponding genes of other proteobacteria. PMID- 16211848 TI - [The effect of sodium salts and pH on hydrogenase activity of the haloalkaliphilic sulfate-reducing bacteria]. AB - Hydrogenase is the main catabolic enzyme of hydrogen-utilizing sulfate-reducing bacteria. In haloalkaliphilic sulfate reducers, hydrogenase, particularly if it is periplasmic, functions at high concentrations of Na+ ions and low concentrations of H+ ions. The hydrogenases of the newly isolated sulfate reducing bacteria Desulfonatronum thiodismutans, D. lacustre, and Desulfonatrovibrio hydrogenovorans exhibit different sensitivity to Na+ ions and remain active at NaCl concentrations between 0 and 4.3 M and NaHCO3 concentrations between 0 and 1.2 M. The hydrogenases of D. lacustre and D. thiodismutans remain active at pH values between 6 and 12. The optimum pH for the hydrogenase of D. thiodismutans is 9.5. The optimum pH for the cytoplasmic and periplasmic hydrogenases of D. lacustre is 10. Thus, the hydrogenases of D. thiodismutans, D. lacustre, and Dv. hydrogenovorans are tolerant to high concentrations of sodium salts and extremely tolerant to high pH values, which makes them unique objects for biochemical studies and biotechnological applications. PMID- 16211849 TI - [Characterization of the lipopolysaccharide from Rahnella aquatilis 1-95]. AB - The lipopolysaccharide from the freshwater bacterium Rahnella aquatilis 1-95 has been isolated and investigated for the first time. The structural components of the lipopolysaccharide molecule: lipid A, core oligosaccharide, and O-specific polysaccharide were isolated by mild acidic hydrolysis. In lipid A, 3 hydroxytetradecanoic and tetradecanoic acids were found to be the predominant fatty acids. In the core oligosaccharide, galactose, arabinose, fucose, and an unidentified component were shown to be the major monosaccharides. The O-specific polysaccharide consists of a regularly repeating trisaccharide unit with the acyl and phosphate following structure: [structure: see text] groups have been shown to be responsible for the toxic and pyrogenic properties of the lipopolysaccharide of R. aquatilis. PMID- 16211850 TI - [Dynamics of the growth and population composition of the mixed cultures of R, S, and M dissociants of Pseudomonas aeruginosa]. AB - The population composition of polycultures of Pseudomonas aeruginosa dissociants (R + M and R + S + M) developing on media with various contents and ratios of nitrogen and phosphorus has been studied. Irrespective of its proportion (10 to 90%) in the inoculum, the R variant accounted for 65 to 84% of the whole population of linear-phase and stationary-phase binary cultures of R and M dissociants, which differ in terms of energy metabolism and nutritional requirements. After prolonged cultivation, the population in the binary culture contained only R cells (100%), which are characterized by minimum requirements with respect to the main biogenic elements. These data agree with the predictive data of model studies and can be attributed to regulation of the population composition of bacterial cultures by trophic factors. It was established that the proportion of M cells, which are distinguished by maximum nutrient requirements and enhanced stability, increased during two developmental stages of the Ps. aeruginosa polycultures (R + M and R + S + M): the lag phase and the decay stage. This result cannot be due to the influence of trophic factors and presumably results from changes in the levels of autoregulatory factors (anabiosis autoinducers) involved in stress resistance and plausibly in the adaptive interconversion of dissociants upon transfer to a new medium (during the lag phase) and under starvation conditions (at the onset of the decay phase). PMID- 16211851 TI - [Conditions favoring differentiation and stabilization of the life cycle of the yeast Pachysolen tannophilus]. AB - Conditions favoring differentiation and stabilization of the life cycle of the yeast Pachysolen tannophilus have been studied. When concentrations of the carbon source in the medium were lower than 100 g/l, it was found to be favorable to the mating of vegetative cells, both haploid and diploid. The addition of nitrogen and sulfur sources to the medium influenced the life phases of haploid cells and partially stabilized the vegetative growth of diploid cells. Enrichment of the nutrient medium with potassium, vitamins, and microelements was shown to be necessary for the formation and maturation of conjugated ascospores. Microelements, vitamins, and phosphorus in excessive amounts activated conjugation but did not provide for the distinct phases of formation of unconjugated asci and spores in the diploid cells. Possible reasons for the unstable diplophase in the yeast P. tannophilus are discussed. PMID- 16211852 TI - [The role of intercellular contacts in the initiation of growth and in the development of a transiently nonculturable state by the cultures of Rhodococcus rhodochrous grown in poor media]. AB - It was found that the growth of Rhodococcus rhodochrous cells in modified Saton's medium strongly depends on the rate of culture agitation in the flask: an agitation at 250 rpm in flasks with baffles stops cell multiplication, whereas slight agitation leads to pronounced culture growth. The growth retardation phenomenon was reversible and did not manifest itself in exponential-phase cultures or when the cells were grown in a rich medium; furthermore, it was not connected with the degree of culture aeration. When agitated at a moderate rate, the bacterial cells formed aggregates in the lag phase, which broke up into single cells in the exponential phase. The inhibitory effect of vigorous agitation was removed by the addition to the medium of the supernatant (SN) of a log-phase culture grown in the same medium with moderate agitation. Vigorous agitation is thought to interfere with the cell contacts, whose establishment is necessary for the development of an R. rhodochrous culture in a poor medium, which occurs in the form of (micro) cryptic growth. When grown in modified Saton's medium, R. rhodochrous cells were capable of transition, in the prolonged stationary phase, to a resting and transiently nonculturable state. Such cells could be resuscitated by incubation in a liquid medium with the addition of the supernatant or the Rpf secreted protein. The formation of transiently nonculturable cells was only possible under the conditions of a considerable agitation rate (250-300 rpm), which prevented secondary (cryptic) growth of the culture. This circumstance indicates the importance of intercellular contacts not only for the initiation of growth but also for the transition of the bacteria to a dormant state. PMID- 16211853 TI - [Adaptive reactions of mycoplasmas in vitro: "viable but unculturable forms" and nanocells of Acholeplasma laidlawii]. AB - The adaptation of Acholeplasma laidlawii to conditions unfavorable for growth has been found to be accompanied by cell transformation into special morphological structures known as ultramicroforms (nanocells). The ratio of the cells of the two morphological types in the population depended on the growth conditions. Nanocells retained viability for a long time under conditions unfavorable for growth and showed resistance to stressors. Reduction in the cell size occurred due to unequal division, which involved the loss of cytoplasmic material. A. laidlawii ultramicroforms (nanocells) were able to restore proliferative activity and to revert to their initial vegetative form; they measured less than 0.2 microm and are the smallest cells known at present. Nanocells formed in vitro under exposure to abiogenic stressors may correspond to the A. laidlawii minibodies observed in infected plants upon exposure to biogenic stressors. The transformation of A. laidlawii cells into ultramicroforms was accompanied by condensation of the nucleoid, a change in the polypeptide spectrum, and a change in the availability of rRNA operons for in vitro amplification. All these changes are indicative of reorganization of the genetic and metabolic systems of mycoplasmas. PMID- 16211854 TI - [Fluorescence microscopic study of the microorganisms treated with chaotropic agents]. AB - The yeasts Saccharomyces cerevisiae and Pichia pastoris and the bacteria Micrococcus luteus, Bacillus subtilis, and Anaerobacter polyendosporus have been treated with the chaotropic agents guanidine hydrochloride and guanidine thiocyanate and certain detergents and studied using fluorescence microscopy. Studies with the use of fluorochromes that can selectively stain nucleic acids (diamidino-2-phenylindole (DAPI), propidium iodide, and acridine orange) show that treatment of the bacterial and yeast cells at 37 degrees C for 3-5 h induces a release of DNA from the cytoplasm and its accumulation in the cellular zone, known as ectoplasm, located between the cell wall and the remainder of the cytoplasm (called endoplasm) in the form of one or several large granules. After treating the cells with the chaotropic agents at 100 degrees C for 5-6 min, the DNA is diffusively distributed over the ectoplasm. The fluorochromes used do not allow the detection of RNA. These findings are in agreement with previous data obtained from electron microscopic study of thin cell sections. After 33 PCR cycles, a considerable portion of DNA leaves the cells; as a result, they show a low level of diffusive fluorescence when stained with DAPI. When endospores of B. subtilis are treated with the chaotropic agents, they become highly permeable to the fluorochromes. Fluorescence microscopic study of such endospores shows that they contain DNA in the central part of their cores. PMID- 16211855 TI - [Methylophaga murata sp. nov.: a haloalkaliphilic aerobic methylotroph from deteriorating marble]. AB - The haloalkaliphilic methylotrophic bacterium (strain Kr3) isolated from material scraped off the deteriorating marble of the Moscow Kremlin masonry has been found to be able to utilize methanol, methylamine, trimethylamine, and fructose as carbon and energy sources. Its cells are gram-negative motile rods multiplying by binary fission. Spores are not produced. The isolate is strictly aerobic and requires vitamin B12 and Na+ ions for growth. It is oxidase- and catalase positive and reduces nitrates to nitrites. Growth occurs at temperatures between 0 and 42 degrees C (with the optimum temperatures being 20-32 degrees C), pH values between 6 and 11 (with the optimum at 8-9), and NaCl concentrations between 0.05 and 3 M (with the optimum at 0.5-1.5 M). The dominant cellular phospholipids are phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. The major cellular fatty acids are palmitic (C16:0), palmitoleic (C16:1), and octadecenoic (C18:1) acids. The major ubiquinone is Q8. The isolate accumulates ectoine and glutamate, as well as a certain amount of sucrose, to function as osmoprotectants and synthesizes an exopolysaccharide composed of carbohydrate and protein components. It is resistant to heating at 70 degrees C, freezing, and drying; utilizes methanol, with the resulting production of formic acid, which is responsible for the marble-degrading activity of the isolate; and implements the 2-keto-3-deoxy-6-phosphogluconate variant of the ribulose monophosphate pathway. The G+C content of its DNA is 44.6 mol%. Based on 16S rRNA gene sequencing and DNA-DNA homology levels (23-41%) with neutrophilic and alkaliphilic methylobacteria from the genus Methylophaga, the isolate has been identified as a new species, Methylophaga murata (VKM B-2303T = NCIMB 13993T). PMID- 16211856 TI - [Elucidation of the taxonomic status of industrial strains of thermophilic lactic acid bacteria by sequencing of 16S rRNA genes]. AB - Phenotypic characteristics and results of PCR tests for the presence of species specific genes indicate that a number of strains of thermophilic lactic acid bacteria previously considered as belonging to Streptococcus thermophilus are actually closely related to enterococci. In the present study, partial (over 500 nucleotides) sequencing of 16S rRNA genes from 12 strains of thermophilic lactic acid bacteria used as starters for manufacturing sour milk products on the territory of the Commonwealth of Independent States (CIS) has been performed. According to the results of the sequencing, seven of the strains have been classified with Enterococcus durans. The earlier classification (based on PCR tests) of two of the strains as S. thermophilus and three of the strains as E. faecium has been confirmed. The data obtained demonstrate that the enterococci E. durans and E. faecium are widely used as thermophilic starters for manufacturing sour milk products on the territory of the CIS. PMID- 16211857 TI - [The construction and monitoring of genetically marked, plasmid-containing, naphthalene-degrading strains in soil]. AB - A genetically marked, plasmid-containing, naphthalene-degrading strain, Pseudomonas putida KT2442(pNF142::TnMod-OTc), has been constructed. The presence of the gfp gene (which codes for green fluorescent protein) and the kanamycin and rifampicin resistance genes in the chromosome of this strain allows the strain's fate in model soil systems to be monitored, whereas a minitransposon, built in naphthalene biodegradation plasmid pNF142, contains the tetracycline resistance gene and makes it possible to follow the horizontal transfer of this plasmid between various bacteria. Plasmid pNF142::TnMod-OTc is stable in strain P. putida KT2442 under nonselective conditions. The maximal specific growth rate of this strain on naphthalene was found to be higher than that of the natural host of plasmid pNF142. When introduced into a model soil system, the genetically marked strain is stable and competitive for 40 days. The transfer of marked plasmid pNF142::TnMod-OTc to natural soil bacteria, predominantly fluorescent pseudomonads, has been detected. PMID- 16211858 TI - [First isolation of the yeast Saccharomyces paradoxus in Western Siberia]. AB - Two ascomycetous yeast strains have been isolated near Novosibirsk from oak exudate. The strains have been identified as Saccharomyces paradoxus Bachinskaya based on the results of biochemical tests. The conspecificity of the isolates with S. paradoxus was confirmed by electrophoretic karyotyping and restriction analysis of the ITS region of its rDNA. This first isolation of S. paradoxus in Siberia provides evidence for the continuity of its natural habitats. PMID- 16211859 TI - [Genetic diversity in pseudomonads associated with cereal cultures infected with basal bacteriosis]. AB - The genetic properties of 45 pseudomonad strains isolated from cereal cultures exhibiting symptoms of basal bacteriosis have been investigated. Considerable genetic diversity has been demonstrated using DNA fingerprints obtained by amplification with REP, ERIC, and BOX primers. Restriction analysis of the 16S 23S internal transcribed spacer (ITS1) allowed the strains to be subdivided into two major groups. In a phylogenetic tree, the ITS1s of these groups fell into two clusters, which also included the ITS1 of Pseudomonas syringae ("Syringae" cluster) and the ITS1 of P. fluorescens, P. tolaasii, P. reactans, P. gingeri, and P. agarici ("Fluorescens" cluster) from the GenBank database. Comparison of the ITS1 divergence levels within the "Fluorescens" cluster suggests expediency of treating P. tolaasii, P. reactans, various P. fluorescens groups, and, possibly, P. gingeri and P. agarici as subspecies of one genospecies. The intragenomic heterogeneity of ITS1s was observed in some of the pseudomonad strains studied. The results of amplification with specific primers and subsequent sequencing of the amplificate suggest the possibility of the presence of a functionally active syrB gene involved in syringomycin biosynthesis in the strains studied. PMID- 16211860 TI - [The saprotrophic bacterial complex in the raised peat bogs of Western Siberia]. AB - The population density of bacteria in peat deposits along the landscape profile of the Vasyugan Marsh has been found to be as high as tens of millions of CFU/g peat. The abundance and diversity of bacteria increased with depth within the peat deposit, correlating with an increasing level of peat degradation. Variations in these parameters with depth and season were greater in peat deposits located in transaccumulative and transitional positions than in the sedge-sphagnum bogs located at the eluvial region of the profile. In the upper 1 m-thick layer of the peat deposits studied, bacilli, represented by five species, dominated, whereas, in the deeper layers, spirilla and myxobacteria prevailed. These bacteria were major degraders of plant polymers. Unlike the bacterial communities found in the peat deposits of European Russia, the dominant taxa in the studied peat deposits of Western Siberia are represented by bacteria resistant to extreme conditions. PMID- 16211861 TI - [Microbiological and isotopic-geochemical investigations of meromictic lakes in Khakasia in winter]. AB - Microbiological and isotopic-geochemical investigations of the brackish meromictic lakes Shira and Shunet were performed in the steppe region of Khakasia in winter. Measurements made with a submersed sensor demonstrated that one-meter ice transmits light in a quantity sufficient for oxygenic and anoxygenic photosynthesis. As in the summer season, in the community of phototrophic bacteria found in Lake Shira, the purple sulfur bacteria Amoebobacter purpureus dominated, whereas, in Lake Shunet, the green sulfur bacteria Pelodictyon luteolum were predominant. Photosynthetic production, measured using the radioisotopic method, was several times lower than that in summer. The rates of sulfate reduction and production and oxidation of methane in the water column and bottom sediments were also lower than those recorded in summer. The process of anaerobic methane oxidation in the sediments was an exception, being more intense in winter than in summer. The data from radioisotopic measurements of the rates of microbial processes correlate well with the results of determination of the isotopic composition of organic and mineral carbon (delta13C) and hydrogen sulfide and sulfate (delta34S) and suggest considerable seasonal variations in the activity of the microbial community in the water bodies investigated. PMID- 16211862 TI - [Aerobic methanotrophic communities in the bottom sediments of Lake Baikal]. AB - The results of the first methodical investigation into the aerobic methanotrophic communities inhabiting the bottom sediments of Lake Baikal are reported. Use of the radioisotopic method revealed methane consumption in 12 10- to 50-cm-long sediment cores. The maximum methane consumption rates (495-737 microl/(dm3 day) were recorded in sediments in the regions of hydrothermal vents and oil and gas occurrence. Methane consumption was most active in the surface layers of the sediments (0-4 cm); it decreased with the sediment depth and became negligible or absent at depths below 20 cm. The number of methanotrophic bacteria usually ranged from 100 to 1000 cells/cm3 of sediment and reached 1 million cells/cm3 in the regions of oil and gas occurrence. The 17 enrichment cultures obtained were represented mainly by morphotype II methanotrophs. Phylogenetic analysis of the enrichment cultures in terms of the amino acid sequence of the alpha subunit of the membrane-bound methane monooxygenase revealed the predominance of methanotrophs of the genus Methylocystis. The results obtained suggest the presence of an active aerobic methanotrophic community in Lake Baikal. PMID- 16211863 TI - [Plasmid rearrangements in Azospirillum brasilense]. PMID- 16211864 TI - Antigen processing and presentation. PMID- 16211865 TI - Antigen recognition and T-cell biology. AB - Despite the wealth of information that has been acquired regarding the way T cells recognize their targets, we are left with far more questions than answers regarding how to manipulate the immune response to better treat cancer patients. Clearly, most patients have a broad repertoire of T cells capable of recognizing their tumor cells. Despite the presence of these tumor reactive T cells and our ability to increase their frequency though vaccination or adoptive transfer, patients still progress. From the T cell side, defects in T cell signaling may account for much of our failure to achieve significant numbers of objective clinical responses. In spite of these negatives, the horizon does remain bright for T cell based immune therapy of cancer. The periodic objective clinical response tells us that immune therapy can work. Now that we know that cancer patients have the capacity to mount immune responses against their tumors, current and future investigations with agents which alter T cell function combined with vaccination or adoptive T cell transfer may help tip the balance towards effective immune therapies. PMID- 16211866 TI - Mechanisms of tumor evasion. AB - The results from in vitro immunological experiments, murine tumor models and patients with cancer clearly demonstrate that tumors have multiple mechanisms to evade the immune response. During the early stages of tumor development malignant cells can be poor stimulators, present poor targets or become resistant to the innate immune response, while at later stages, progressively growing tumors impair the adaptive immune response by blocking the maturation and function of antigen presenting cells and causing alterations in T cell signal transduction and function. Preliminary results also suggest a correlation between some of these changes and an increased metastatic potential of the tumor cells, a diminished response to immunotherapy, and poor prognosis. Carefully coordinated basic research studies and clinical immunotherapy trials will be required to fully determine the impact on the outcome of the disease and the response to treatment. However, understanding the mechanisms used by tumor cells to evade the immune system could result in new therapeutic approaches for preventing and/or reversing these immune alterations and have the potential of improving the current results of immunotherapy trials. PMID- 16211867 TI - Tumor antigens and tumor antigen discovery. PMID- 16211868 TI - Peptide vaccines against cancer. PMID- 16211869 TI - DNA vaccination in immunotherapy of cancer. PMID- 16211870 TI - Antibody inducing polyvalent cancer vaccines. AB - The great majority of cancer patients can initially be rendered free of detectable disease by surgery and/or chemotherapy. Adjuvant chemotherapy or radiation therapy are generally only minimally beneficial, so there is real need for additional methods of eliminating residual circulating cancer cells and micrometastases. This is the ideal setting for treatment with a cancer vaccine. The immune response induced is critically dependent on the antigenic epitope and vaccine design. For antibody induction there is one best vaccine design, conjugation of the antigen to an immunogenic protein such as KLH and the use of a potent adjuvant such as the saponins QS-21 and GPI-0100. This approach alone induced strong antibody responses against the glycolipids GM2, fucosyl GM1 and globo H and the mucin backbone MUC1, and cancer cells expressing these antigens. Other antigens required additional modifications to augment relevant immunogenicity. GD2 and GD3 lactones and N-propionylated polysialic acid were significantly more effective at inducing antibodies against tumor cells than the unmodified antigens. Tn, sTn and TF trimers (clusters) were significantly more effective than the monomers at inducing antibodies reactive with the cancer cell surface. The optimal approach for Le(Y), KSA, PSMA, and CA125 (MUC16) remains to be determined. Antibodies are ideally suited for eradicating pathogens from the bloodstream and from early tissue invasion. Passively administered and vaccine induced antibodies have accomplished this, eliminating circulating tumor cells and systemic or intraperitoneal micrometastases in a variety of preclinical models, so antibody-inducing vaccines offer real promise in the adjuvant setting. Polyvalent vaccines will probably be required due to tumor cell heterogeneity, heterogeneity of the human immune response and the correlation between overall antibody titer against tumor cells and antibody effector mechanisms. Over the next several years, Phase II clinical trials designed to determine the clinical impact of polyvalent conjugate vaccines will be initiated in the adjuvant setting in patients with SCLC and several epithelial cancers. PMID- 16211872 TI - Undefined-antigen vaccines. AB - Our knowledge of the immune system and how it interacts with tumor cells continues to grow. With each advance in basic science comes a new opportunity to develop an effective treatment strategy. Many such opportunities have arisen in the past few decades and this chapter has attempted to describe how these new advances have been combined with a variety of undefined cellular antigen preparations in an attempt to develop effective cancer vaccines. None of the strategies described in this chapter have been sufficiently effective to become part of standard therapy. However, the approaches tested have generally been well tolerated by patients with advanced cancer and the evidence of immunologic activity and examples of impressive clinical activity in a wide variety of malignancies, suggests that these strategies can be the building blocks upon which new advances are added and effective treatments developed. PMID- 16211871 TI - Dendritic cell-based vaccines for cancer therapy. PMID- 16211873 TI - Cancer vaccines in combination with multimodality therapy. PMID- 16211874 TI - Cytokine therapy for cancer: antigen presentation. PMID- 16211876 TI - Adoptive cellular immunotherapy of cancer: a three-signal paradigm for translating recent developments into improved treatment strategies. PMID- 16211875 TI - Tinkering with nature: the tale of optimizing peptide based cancer vaccines. PMID- 16211877 TI - Clinical trial designs for therapeutic cancer vaccines. AB - Therapeutic cancer vaccines have characteristics that require a new paradigm for phase I and phase II clinical development. Effective development plans may take advantage of some of the following observations: Dose ranging safety trials are not appropriate for many cancer vaccines. Dose ranging trials to establish an optimal biologic dose are often not practical. We have presented an efficient design of Korn et al. (4) to identify an immunogenic dose. Vaccine efficacy can be efficiently evaluated with tumor response as endpoint utilizing a two stage design with only 9 patients in the first stage. If no partial or complete responses are observed in the initial 9 patients, accrual to the trial is terminated. Optimization of vaccine delivery by comparing results of single arm phase II studies using immunological response as endpoint is problematic because of assay variation and potential non-comparability of patients in different studies. Randomized screening studies can be used to efficiently optimize vaccine immunogenicity. Efficiency in use of patients depends on having assay variation and inter-patient variability small relative to the difference in immunogenicity to be detected. Phase II studies using time to progression as endpoint are most interpretable if they employ randomized designs with a no-vaccine control group. Such designs may use an inflated type 1 error rate, and need not be prohibitively large if patients with rapidly progressive disease are studied. Interim monitoring plans may effectively limit the size of the trials by terminating accrual early when results are not consistent with the targeted improvement. PMID- 16211878 TI - Clinical trial design and regulatory issues for therapeutic cancer vaccines. PMID- 16211879 TI - Immune monitoring. AB - A wide array of immunologic tests are available for immune monitoring in cancer vaccine trials, and the number of novel assays and technical modifications continues to burgeon. Because only a small fraction of all proposed vaccine trials tested in phase I-II trials, for practical reasons, will ultimately move forward to be tested in phase III trials, there must be a system of establishing the most promising immunization strategies. This evaluation of cancer vaccine will require standardization of the immune assays and statistical methods used in immunologic monitoring. Furthermore, the use of a systematic approach to evaluating and adopting novel technologies for immunologic assessment would likely lead to timely implementation of more reliable, practical and cost effective methods of immune. It should be the goal and expectation that this rational approach to immune monitoring will allow the critical appraisal of the most promising vaccine candidates in the context of pivotal, multi-center trials. PMID- 16211880 TI - Anatomy and physiology of the leptomeninges and CSF space. AB - The arachnoid membrane and pia mater are the two membranous layers that comprise the leptomeninges. Cerebrospinal fluid is made within the ventricular system by cells of the choroid plexus and ependyma. This chapter describes in detail the normal anatomic structure and physiologic interactions of the cerebrospinal fluid and leptomeningeal space that are critical to our understanding and treatment of leptomeningeal metastases. PMID- 16211881 TI - Leptomeningeal neoplasia: epidemiology, clinical presentation, CSF analysis and diagnostic imaging. AB - The incidence of leptomeningeal metastasis over the past several decades has increased among solid tumor patients and decerased in patients with hematologic malignancies. Improvements in systemic therapies are likely responsible for both changes; solid tumor patients are living longer and, threfore, are at higher risk to develop leptomeningeal tumors while patients with hematologic malignancy have benefitted from more aggressive central nervous system prophylaxis. Regardless, both types of patients present with symptoms referrable to multiple levels of the central nervous system and a careful dignostic approach incorporating cerebrospinal fluid studies and appropriate neuroimaging is critical. PMID- 16211882 TI - Leptomeningeal metastases from solid tumors (meningeal carcinomatosis). AB - Leptomeningeal metastases (LM) are increasingly recognized as a devastating complication of solid tumors. Improved treatment of primary malignancy and advances in diagnostic imaging have led to an apparent increase in the number of patients diagnosed with LM. Unfortunately, therapeutic options remain limited. Radiotherapy is used to treat bulky tumor and provide symptomatic relief. Intrathecal chemotherapy benefits a selected subset of patients. The challenge to the future is to delineate the molecular mechanisms underlying LM and to develop novel therapeutic or prophylactic modalities to combat LM. PMID- 16211883 TI - Leptomeningeal metastases from leukemias and lymphomas. AB - Leptomeningeal dissemination of lymphoma and leukemia differs from that of solid tumors in a number of clinically important aspects. Specific histologic variants of lymphoma and leukemia have such a high incidence of cerebrospinal fluid (CSF) dissemination that assessing CSF cytology at diagnosis is crucial and prophylactic therapy of the CSF compartment is required. Furthermore, while the overall prognosis for patients with leptomeningeal metastases from leukemia and lymphoma is similar to solid tumors, selected patients have excellent response to therapy and attain durable remission. Therefore, aggressive treatment is warranted. PMID- 16211884 TI - Leptomeningeal metastasis of primary central nervous system (CNS) neoplasms. AB - Leptomeningeal dissemination of primary CNS tumors varies widely by histologic subtype. In certain tumors including medulloblastoma, ependymoma, germ cell tumors, and primary CNS lymphoma, seeding of the cerebrospinal fluid space is a critical factor in determining stage, prognosis and appropriate therapy. Other tumor types, such as glioma, may have radiographic evidence of leptomeningeal metastases without clear impact on prognosis or therapy. PMID- 16211885 TI - Leptomeningeal cancer in the pediatric patient. AB - The treatment and prophylaxis of leptomeningeal leukemia and lymphoma in children has dramatically improved disease control and long-term survival. However, the treatment of other leptomeningeal cancers has been less successful and the neurologic morbidity associated with central nervous system-directed therapy has a significant long-term impact on quality of life. Further research is critical to identify new therapeutic strategies for children with or at high risk for leptomeningeal cancer. PMID- 16211886 TI - Neurosurgical interventions for leptomeningeal tumor. AB - Nonsurgical modalities remain the keystone to treatment and control of leptomeningeal tumor. However, neurosurgical consultation is often required for placement of Ommaya reservoirs and ventriculoperitoneal shunts. In this chapter, the utility of these devices, as well as their common complications will be presented. The aim will be to familiarize non-surgical providers with the indications for and complications associated with these neurosurgical procedures as a component of the adjuvant treatment of leptomeningeal disease. PMID- 16211887 TI - Current treatment of leptomeningeal metastases: systemic chemotherapy, intrathecal chemotherapy and symptom management. AB - Treatment of leptomeningeal metastases is multifaceted and includes symptomatic therapy, intrathecal and systemic chemotherapy, and radiotherapy. As the majority of patients have widespread incurable systemic tumor, treatment is predominantly palliative; however, some patients with leukemia, lymphoma or breast cancer may have prolonged remissions and the possibility of cure. PMID- 16211888 TI - Radiation therapy for leptomeningeal cancer. AB - Radiotherapy has multiple roles in the treatment of leptomeningeal cancer. While it is uncommon for patients to experience regression of neurologic deficits due to leptomeningeal cancer, focal radiotherapy often provides significant palliation of pain, increased intracranial pressure and other focal symptoms. Focal radiotherapy may also be used to eliminate blockages of cerebrospinal fluid (CSF) and allow for safe administration of intrathecal chemotherapy. Craniospinal irradiation (CSI) is most often used as prophylaxis for patients at high risk of leptomeningeal tumor dissemination, but may result in symptom palliation and prolonged disease control for patients with active leptomeningeal tumor. PMID- 16211889 TI - Animal models of leptomeningeal cancer. AB - Animal models are a critical tool for our understanding of pathogenic mechanisms and the development of therapeutic strategies. Since the 1970's, numerous syngeneic and allogeneic rodent models of leptomeningeal cancer have been developed; in this chapter, we present representative models and discuss their clinical and translational implications. PMID- 16211890 TI - Improving the outcome of patients with leptomeningeal cancer: new clinical trials and experimental therapies. AB - Current therapy for leptomeningeal metastases is predominantly palliative. In an effort to improve disease control and patient outcome, new strategies are being developed to target the cerebrospinal space. These include new intrathecal formulations of systemic chemotherapy as well as the development of radiolabeled immunoconjugates and antitumor antibodies. Furthermore, there is debate as to the optimal strategy of drug delivery for leptomeningeal tumor. PMID- 16211891 TI - [Importance of estimation of unconjugated bilirubin not linked to albumin in the new born and premature jaundiced baby]. PMID- 16211892 TI - [Results and duration of treatment by light therapy of jaundice in new born and premature infants. Effects of light on unbound bilirubin]. PMID- 16211893 TI - [Toxicity of oxygen on lung tissue in the new born]. PMID- 16211894 TI - [Technics of detection of urinary infection in children]. PMID- 16211895 TI - [Centrifugal annular erythema of Colcott Fox type (erythema gyratum perstans)]. PMID- 16211896 TI - [Treatment and rehabilitation of myopathies linked to sex in relation to prognosis]. PMID- 16211897 TI - [Children "healthy carriers" of pathogenic Escherichia coli in nurseries]. PMID- 16211898 TI - [Mesenteric cysts and cysts of the mesocolon in the child. Report of three cases of cystic lymphangioma]. PMID- 16211899 TI - [Association of pigmented retinopathy and Duchenne's disease in one family]. PMID- 16211900 TI - [The risk of demedicalisation of maternity homes, clinics and centres]. PMID- 16211901 TI - [Intra uterine major dwarfism with dysmorphia and severe encephalopathy. Bird head dwarfism (Virchow-Seckel type)]. PMID- 16211902 TI - [Arachnoid cyst of the IIIrd ventricle and early puberty. Clinical and neuroradiological study and treatment with progestagens]. PMID- 16211903 TI - [Association of congenital hypoaldosteronism and steatorrhoea]. PMID- 16211904 TI - [Familial hydranencephaly]. PMID- 16211905 TI - [Necrotising enterocolitis in the new born. Comments in 64 cases]. PMID- 16211906 TI - [Amyotrophic paralysis of the shoulder]. PMID- 16211907 TI - [Oesophageal atresia (1972-1976)]. PMID- 16211908 TI - [Macroscopic hematuria during treatment with phenylbutazone derivatives. Positive lymphocyte transformation test for these drugs. Two cases in children]. PMID- 16211909 TI - [Xanthine-oxidase deficiency (congenital xanthinuria). Family study]. PMID- 16211910 TI - [Myotonic chondrodystrophy (or Schwartz-Jampel syndrome). Study of siblings and review of the literature]. PMID- 16211911 TI - [Fabry's disease in children. Clinical and biological study of one family. Structure and ultrastructure of the kidney in a hemizygote and a heterozygote]. PMID- 16211912 TI - [Multiple endocrine adenomas type II b. (Syndrome "Marfanoid aspect, multiple mucosal neuromatosis, medullary carcinoma of the thyroid, pheochromocytoma"). Family study and review of the literature]. PMID- 16211913 TI - [Sawyer James MacLeod syndrome in the child]. PMID- 16211914 TI - [Neonatal leucinosis with favourable course. Therapeutic problems]. PMID- 16211915 TI - [Isochromosome long arm 18]. PMID- 16211916 TI - [Pharmacokinetic and clinical study of cefazoline in children]. PMID- 16211917 TI - [Chronic juvenile rheumatoid arthritis: introduction]. PMID- 16211918 TI - [Enquiry concerning the incidence of chronic juvenile rheumatoid arthritis in France in 1972]. PMID- 16211919 TI - [Clinical forms of chronic arthritis in children]. PMID- 16211920 TI - [Course and prognosis of systemic forms of chronic juvenile rheumatoid arthritis]. PMID- 16211922 TI - [Serous involvement during chronic arthritis in children]. PMID- 16211921 TI - [Course and prognosis of the non systemic forms of chronic juvenile arthritis]. PMID- 16211923 TI - [Modern technics of clinical neurophysiology: introduction]. PMID- 16211924 TI - [The place of electromyography in the diagnosis of myopathy in children]. PMID- 16211925 TI - [EMG study of swallowing in the young child]. PMID- 16211926 TI - [149 cases of convulsions during the first 18 months of life. Statistical analysis and electro-clinical correlations]. PMID- 16211927 TI - [Electro-clinical physionomy of a population of children followed up for more than ten years for convulsions]. PMID- 16211928 TI - [The value of E.E.G changes in head injuries in childhood]. PMID- 16211929 TI - [The evoked potential, cortical manifestation of sensory perception]. PMID- 16211930 TI - [Visual evoked potentials in children. Interest and indications]. PMID- 16211931 TI - [Enquiry on the diagnostic and prognostic value of auditory evoked potentials in children]. PMID- 16211932 TI - [A cause of post-natal anemia, urinary infection]. PMID- 16211933 TI - [Subacute necrotising hepatitis with repeated attacks of fever]. PMID- 16211935 TI - [Suction difficulties in a two month old child confirmed by E.M.G]. PMID- 16211934 TI - [Postoperative bacterial jaundice in a child]. PMID- 16211936 TI - [Neuro-meningeal cryptococcosis in the child. Apropos of 2 cases]. PMID- 16211938 TI - [Child abandoned by the doctor]. PMID- 16211937 TI - [Trials with maternal milk in the low birth weight infant (30 cases)]. PMID- 16211939 TI - [Rare complications of rheumatoid purpura. Pathogenic discussion]. PMID- 16211940 TI - [Attempt at objective evaluation of neonatal infection]. PMID- 16211941 TI - [Plasma red cell and urinary magnesium in the healthy new-born child during the first three weeks of life]. PMID- 16211942 TI - [Skin angioma and heart failure]. PMID- 16211943 TI - [An exceptional cause of neonatal intestinal obstruction: pre-duodenal portal vein. Report of two cases]. PMID- 16211945 TI - [Severe neonatal hypoglycemia due to pancreatic nesidioblastosis. Methods of diagnosis and treatment]. PMID- 16211944 TI - [Salt losing syndrome in the new-born. New diagnostic approach. Report of our experience between 1960 and 1976]. PMID- 16211946 TI - [Delayed growth with raised circulating growth hormone and incapacity to produce somatomedin (dwarfism of Laron type)]. PMID- 16211947 TI - [Multinodular goiter and hyperthyroidism in children]. PMID- 16211948 TI - [Anorexia in hospital--1965-1975]. PMID- 16211949 TI - [A new rapid and reliable test for demonstration of streptococcal anti-exoenzyme antibodies]. PMID- 16211950 TI - [Study of early neonatal mortality in the Ille et Vilaine department (1972-1973 1974)]. PMID- 16211952 TI - [Breast-feeding in Brittany]. PMID- 16211951 TI - [The fate of new born admitted to hospital for prematurity and resuscitation in Rennes]. PMID- 16211953 TI - [20 cases of trisomy 18. Sex-ratio in relation to age of the mother]. PMID- 16211954 TI - [Congenital lipomatous hypoplasia of the exocrine pancreas in a child. Report of a new case]. PMID- 16211955 TI - [Aorto-renal vascular lesions during neurofibromatosis]. PMID- 16211956 TI - [Recent progress in biochemical and histological research during Morquio's disease. Report of three new cases]. PMID- 16211957 TI - [Deafness in children and the use of an hearing aid]. PMID- 16211958 TI - [Indications for stool culture and antibiotics in the treatment as an out patient, of diarrhea due to acute gastro-enteritis in infants]. PMID- 16211959 TI - [The treatment of enuresis by a group educational technique]. PMID- 16211960 TI - [The opinion of parents on the risks of accidental poisoning in their children: enquiry]. PMID- 16211961 TI - [Thoracic dystrophy and nephropathy]. PMID- 16211962 TI - [A case of cystic lymphangioma of the mesentery in the child]. PMID- 16211963 TI - [Glomus tumour with neurological signs. First observation in a child]. PMID- 16211964 TI - [Primary IgA deficiency (pure or in association). Report of 15 cases]. PMID- 16211965 TI - [Secretory IgA in children with urinary infection]. PMID- 16211966 TI - [Interest of estimation of immunoglobulins in pediatrics]. PMID- 16211967 TI - [Congenital non-bullous ichthyosiforme erythroderma. Two brothers with a particularly severe form]. PMID- 16211968 TI - [530 cases of poisoning at the Enfants Malades Hospital]. PMID- 16211969 TI - [Abdominal pain and spasmophilia in children]. PMID- 16211971 TI - [Acromegalogigantism]. PMID- 16211970 TI - [Congenital bullous epidermolysis. Report of 8 cases]. PMID- 16211972 TI - [Plethysmography in the child]. PMID- 16211973 TI - [Dermatomyositis in children. Report of 5 cases]. PMID- 16211974 TI - [Presentations of trichinosis in children. A recent epidemic]. PMID- 16211975 TI - [Benign rheumatoid nodules. Report of one case]. PMID- 16211976 TI - [Epidermoid cysts of the spleen. Report of one case]. PMID- 16211977 TI - [Foreign bodies within the bronchus in children]. PMID- 16211978 TI - [Intestinal obstruction in the new born]. PMID- 16211979 TI - [Acute poisoning by colchicine in children. Report of a 4 cases]. PMID- 16211980 TI - [Acute lead poisoning during the perinatal period. A case of encephalopathy in a six week old child treated by peritoneal dialysis]. PMID- 16211981 TI - [Digestive intolerance to cow's milk protein in the infant. Clinical study of 15 cases]. PMID- 16211982 TI - [Cystinosis in the child. Semiological aspects and course in 3 cases]. PMID- 16211983 TI - [Interstitial pneumonia due to cholesterol in the child]. PMID- 16211984 TI - [Subacute encephalopathy of early onset during treatment of relapse of lymphoblastosis in a young infant]. PMID- 16211985 TI - [Study of an anxiolytic drug in infancy: clobazam]. PMID- 16211986 TI - [Regarding postgraduate education in the School of Medical Sciences]. PMID- 16211987 TI - [Risk factors associated to upper gastrointestinal bleeding in non intensive care units]. AB - Upper gastrointestinal bleeding--UGB-, as a complication, is well studied at intensive care units (ICU), but is less known in non ICU settings. OBJECTIVES: To determine incidence and risk factors of this entity at clinical hospitalization. MATERIALS AND METHODS: A case-control study of patients with gastric ulcer disease diagnosed by endoscopy who presented with melena and hematemesis. Ten controls were taken for each case, matching sex, age and prophylaxis for gastric hemorrhage. Demographic data and other know risks factors were analyzed. RESULT: We found ten bleeding case among 35070 discharges (incidence: 2.8/10000 discharges). Mortality was not increased but the number of transfusion was higher in the bleeding group. We found an assocciation betwen UGB and systemic inflammatory response syndrome--SIRS-(OR: 9.22 IC 95% 2.98-28.17) and diabetes (OR: 7.8 IC 95% 2.3-26.8). The rest of the factors studied did not rich a statistical significance. CONCLUSIONS: UGB during clinical hospitalization is a rare complication that requires an increased staying at hospital and a great number of transfusions. It may be probably associated in a positive way with diabetes and SIRS. PMID- 16211988 TI - Renoprotective effect of high periprocedural doses of oral N-acetylcysteine in patients scheduled to undergo a same-day angiography. AB - BACKGROUND: Few studies that have assessed the effect of abbreviated oral N acetylcysteine (NAC) regimens in radiocontrast-induced nephropathy (RCIN) yield mixed results. OBJECTIVE: To evaluate the renoprotective effect of high periprocedural oral doses (HPOD) of NAC in patients with chronic renal impairment undergoing a same-day angiography. METHODS: Sixty one patients with renal impaired function scheduled to undergo a same-day angiography were randomly assigned to NAC 1200 mg orally 3 hours before and 3 after the procedure, or a placebo. All patients received 0.9% saline intravenous. RCIN was defined as an increase in SCC > 0.5 mg/dl 48 hours after the procedure. RESULTS: The mean baseline SCC for all patients was 1.44 +/- 0.42 mg/dl. A significant difference in SCC change at 48 hours after the angiography was found (-0.07 mg/dl NAC, 0.09 mg/dl placebo, P = 0.04). RCIN occurred in 1 (3%) patient of NAC group and in 2 (7.1%) patients of placebo group (P = 0.59). Adverse effects were similar in both groups. CONCLUSIONS: In patients with mild renal impairment patients undergoing angiographic procedures, HPOD of NAC were more effective than placebo in preventing SCC change 48 hours. A non significant benefit in RCIN incidence was found. PMID- 16211989 TI - [Body training and insulin resistance in two rural communities of Cordoba - Argentina]. AB - From the AF, the smaller risk is controversial. The influence of the general and environmental components on IR is not know in a satisfactory way. OBJECTIVES: The influence of AF on the prevail IR in two rural communities of Cordoba with different fenotipical configuration and IR risk. MATERIAL AND METHODS: Transverse study (survey, clinical and biochemical checkup) over 1143 people of Oncativo community (ON) and Dean Funes (DF), inclusion: n = 673 (30-60 years old) Normal ECG, no diabetics. Sorts AF according to the spend of calories (Kcal/sem): a) sedentary (< 500), b) low (5001-1000), c) regular (1001-2500), and d) High (> 2500). IR is HOMA > 2.5; where [HOMA = (Glucemia * Insulina)/ 4.5]. RESULTS: AF: 31.4% sedentary, 20.2% low, 26.3% regular and 22.1% high. 16.5% IR in ON opposite 34.8% en DF (P < 0.001), IR in ON 8.2% with AF high opposite 18.8% the rest (P = 0.032), 36.8% and 34.2% like in DF (P = 0.67). With the adjusted results for age and community it was found less IMC (P = 0.002) and waist circumference (P = 0.042) than the most category of AF. The blood pressure had not a meaningful change, lipid profile or HOMA depending of AF. CONCLUSION: if a benefit of AF over IR exists, it will be conditioned for genetics and environmental primaries determiners. PMID- 16211990 TI - [Proliferation and apoptosis in tongue epithelium of mice bearing salivary tumors induced by dimetilbenzantraceno (DMBA) and modulated by dietary lipids]. AB - INTRODUCTION: According to the concept of field defects during the carcinogenesis process, excessive epithelial proliferation/apoptosis may exist in areas near tumors. Proliferation or apoptosis could be modified by dietary lipids. PURPOSE: The present study was designed to analyze proliferation and apoptosis in tongue epithelium of mice fed diets based on different lipids followed by induction of salivary tumors with DMBA. MATERIALS AND METHODS: Forty-five days after weaning, ten BALB/c mice were assigned to two diets: corn oil (CO) and fish oil (cod liver, FO). Two weeks later, DMBA was injected in the submandibular area. Animals were sacrificed at the 13th post-injection week. Samples of tongue were fixed in formalin-ethanol and immunohistochemically stained for proliferation (Ki-67) and apoptosis (Bax). By light microscopy, the number of nuclei positive for these markers were counted out of three-hundred total interphase cells both in dorsal and in ventral tongue surfaces. Results were analyzed through Analysis of Variance and t Test. RESULTS: Cell proliferation was greater in dorsal than in ventral tongue surfaces (p < 0.0001) with no diet difference. Apoptosis was significantly greater in mice fed FO than CO, particularly in tongue dorsal epithelia (p < 0.018). CONCLUSIONS: This study shows that FO diet induces higher levels of apoptosis in tongue epithelia suggesting a tissue defensive mechanism when exposed to a carcinogenic-tumoral agent. PMID- 16211991 TI - [Assessment of health and record of disease in the promotion of health oral]. AB - BACKGROUND: At present, epidemiologic dentistry has numerous indexes to measure illness such as DMFT, dmft and S-OHI which are broadly used for illness diagnosis in different population groups. On the other hand, the index INSAO proposed differs from the approach focused on the disease itself. INSAO offers a parameter to measure the degree of health of teeth and gums and thus provides the patient with the information about their health. OBJECTIVE: The aim of this study is to analyze the usefulness of the information provided by the traditional indexes and those recorded by INSAO, to the oral health evaluation in rural schools of the north of Cordoba, Argentina. MATERIAL AND METHODS: A cross-sectional study was carried out in school children, aged 5 to 14 attending 8 rural primary schools. Data was collected from a clinical examination. Caries experience was measured according WHO indications. The dmf-t and DMF-T indexes were determined. Oral hygiene was evaluated by means S-OHI. The total amount of teeth present in the mouth, sound teeth number, and the state of the gum were registered to built INSAO index. RESULTS: Inverse correspondence is observed between dental health level through INSAO and the disease through proportion of permanent and temporary teeth with cavities active recorded by dmf-t and DMF-T indexes. CONCLUSION: The application of INSAO is presented as a valuable instrument in the promotion of oral health in communities that belong to excluded social groups due to their special characteristics. PMID- 16211993 TI - [Permanence of the piamontese culture on food practices in third generation descendants in Piamonte (Province of Santa Fe, Argentina)]. AB - This paper describes the influence of the piemontese culture on food practices of students living in Piamonte, Santa Fe, Argentina. Food practices of 96 students with Piamontese ancestry (PA) (n = 57) and without Piamontese ancestry (No-PA) (n = 39) were studied along 2002 using a self-administered questionnare. Data were analysed by Chi square test, Fisher's exact test, multiple correspondance analysis and logistic regression. Consumption of bagna cauda (p < 0.05) and polenta (p < 0.1) were higher among PA. Differences on the elaboration of polenta and pasta were found: PA add them cheese (p < 0.05) and cream (p < 0.05) whereas no-PA make use of meat (p < 0.05) and tomato sauce (p < 0.05), respectively. The frequency of consumption of traditional Piamontese meals and the role of the mother in the purchase, the elaboration and the serving of the food were similar on both groups. In conclusion, food practices of Piamontese's descendants recall the food culture of their ancestry with some reasonable adaptations to the local context. PMID- 16211992 TI - [Oral histology and embryology clinically integrated as a teaching tool in an undergraduated dentistry course of studies]. AB - The teaching of Oral Histology and Embryology clinically integrated was designed as a pilot experience to be developed during the 2005 academic year at the Division of Histology and Embryology (Chair "A") of the National University of Cordoba School of Dentistry. This experience, in which the members of the faculty of the Department of Clinical and Basic Sciences have an active participation, is based on a systemic conception of the learning-teaching process and on the recommendations made by the OPS/OMS. This approach will allow us to optimize the quality of our undergraduate programs through better teacher training and the gradual integration of basic and clinical sciences. Our aim is to provide a better education with clinical relevance in basic sciences and scientific basis in clinical assistance. PMID- 16211994 TI - [The Library of Medicine of Cordoba, witness and protagonist of knowledge from the end of the 19th Century to the beginning of 21st Century]. AB - In the nineteenth century, was founded the Medicine's Library of the National University Faculty of Medicine, center of knowledge, new ideas and teaching. The history library sector hold the valuable inheritance that represent the collection of Teachers Doctors, Pablo Mirizzi, Temistocles Castellano, Pedro Ara, Jose F. Verna, Juan Martin Allende located with the books and thesis of the XIX century and beginning of XX century. In 1986 it integrates the National Health Science Information Network and also, the Latino American and the Caribbean Health Science Information Network. The health libraries are in front of a new paradigm. The technologies of information and communication require new methods for administration of the information's resources and services, to answer the society information needs in the knowledge age. The traditional and virtual library is one unit that complement sources in different supports, electronic publications and interactive networks. It is the nexus between the scientific inheritance and the society, and it's mission is: Give local and virtual information's services and management the information's resources, basis of the knowledge society, support of teaching, investigation and extension of the Medical Sciences Faculty of the Cordoba National University. PMID- 16211995 TI - [Medicine and art. Comment about a gallery of paintings]. PMID- 16211996 TI - [Castleman disease]. AB - A 66 years female, who was since last year under astenia, arthralgias, pimply lesions in spread plates and tests showing eritrosedimentation over 100 mm, anemi, leucocitosis with neutrofilia, policlonal hypergammaglobulinemia, slight proteinuria and IgE on 900. This patient was sporadically treated with corticoids. When made the medical consult had lost 34lb., was under anorexy, as well as dyspepsia. Hemoglobyn 6.9 gr/dl, leucocytes 20000/mm3, neutrofils at 90%, proteinogram the same as former, with hypoalbuminemia. She was taking prednisona, 16 mg/day. When examined showed depress of conscience, astenia, and dermic lesions already quoted. 4 cm nonpainful right axillary adenopaty adhered to deep planes. Medulogram with increased iron, hyperegenerative. Ganglionar biopsia: linfoid hyperplasic process linked to inmune response. Toracoabdominal tomography with adenomegalia in torax and retroperitoneo. Skin biopsia: neutrofilic vasculitis. The patient suspends the 16 mg of prednisona and fever as well as generalized adenopatias come up. After laying aside other ethiologies, and understanding as Castleman Multicentric disease, it is started to supply prednisona 1 mg/kg of weight with a clinical and biochemical fast and outstanding response. After 7 months it was progressively suspended the esteroids and 60 days later, the process fall back; for that, corticoids are restarted, with a good evolution. The illness of Castleman although it is not very frequent, it should be considered as differential diagnosis in those clinical cases that are accompanied with important general commitment, linphadenopaties and respons to steroid therapy. PMID- 16211997 TI - [Brown tumor in hyperparathyroidism secondary to chronic renal failure]. AB - Brown tumor (BT) is an uncommon condition that represents the terminal stage of the cystic osteitis fibrosa and have been increasingly reported in hyperparathyroidism secondary to renal failure, due to the increase of survival in patient with hemodialysis. The fine needle aspiration diagnosis is of great importance in the recognition of the BT, although it can be difficult to distinguish it of lesions as the aneurysmal bone cyst and giant-cell tumor. We describe the case of 20-year-old female with chronic renal failure undergoing hemodialysis during six years. Both x-rays and computer tomography revealed a tumor in head of right humerus and lytic images in scapula of the same side, clavicles and ribs. The patient was subjected to a fine needle aspiration biopsy of the tumor of humerus head and the sample was processed with the habitual technique of inclusion in paraffin and stained with hematoxilina and eosina. Histological preparations showed several multinucleate giant cells and spindly or fibrillary cells, feature that was pointed out as compatible, in a context of secondary hyperparathyroidism to chronic renal failure, with a BT. We consider that the radiological and tomographyc finds, besides the history of chronic renal failure with a long history of hemodialysis, were enough to link, with great approach, the histopathology with the diagnosis of BT. PMID- 16211998 TI - [Thoracoscopic resection of mediastinal hypersecretant parathyroid gland]. AB - In general experienced hands, the inferior parathyroid glands, localized in the thymus or low extrathymus are the main cause or surgical therapeutic failure. When they could not be approached through cervicotomy, or when they were diagnosed after surgery the sternotomy or the angiographic ablation were chosen as treatment. The limited number of thoracoscopic approach whit good results published so far, moved us to use this method. MATERIAL AND METHOD: Patient of 49 years with a 7 year secondary hiperprathyroidism due to CRI. After a sub total parathyroidectomy with bilateral thimectomy, the patient did not present clinical or laboratory improvement. Through a thoracic centellogram sixth MIBI, NMR and CAT, a tumor in the middle mediastinum was fond. The mediastinal structures have been easily identified through a left thoracoscopic approach. At the level of the aortopulmonary window, a tumoral mass is located and, with a simple dissection, et is easily extracted in bag. The biopsy through freezing confirms the parathyroid etiology of the gland. RESULT: Clinical and laboratory evolution has been favorable, with hospital discharged at the 3 day, and being asymptomatic after two years. CONCLUSION: The hipersecretant parathyroid glands, located in middle mediastinum, that can not be approach through cervicotomy, can be successfully approach through thoracoscopic technique. PMID- 16211999 TI - [Neuroendocrine carcinoma of the prostate]. AB - Small cell of undifferenciated tumors are present in almost all organs, and it impose the need of performing a differential diagnosis between undifferenciated tumors with residual differentiation according to the type of organ, and the carcinoma of small cells of neuroendocrine origin. The concept of neuroendocrine differentiation (NED) in the prostatic adenocarcinoma has reached considerable attention due to its prognostic and therapeutic implies. Here it is presented a new neuroendocrine prostatic carcinoma case taking care of its hystopathologic diagnosis and evolution. PMID- 16212000 TI - [Foie gras: mea culpa]. PMID- 16212001 TI - [Celiac disease in disguise]. AB - Contrarily to a widely prevalent opinion, celiac disease frequently affects adults, and only rarely reveals itself by the classical triad of diarrhea--weight loss--nutritional deficiency. In addition to isolated deficiencies, most frequently iron and calcium-vitamin D, celiac disease is commonly associated with atypical, sometimes very commonplace manifestations, such as abdominal symptoms reminiscent of those of irritable bowel syndrome, or type I diabetes. The diagnostic process is now made easier by the availability of antitransglutaminase antibodies dosage, a simple, trustworthy, sensitive and specific test. This review article discusses the many clinical pictures which should prompt the clinician to rule out celiac disease, and provides practical guidelines as to the use and interpretation of serologic tests. PMID- 16212002 TI - [New aspect in the treatment of chronic hepatis B]. AB - The pegylated interferon is now the first choice of treatment for patients without a counter-indication. The association of this treatment with lamivudine does not increase the effectiveness. For patients non-responders to the PEG-IFN or presenting counter-indications, the long-term administration of lamivudine is limited by the frequent appearance of mutations, so that escape from the treatment requires the use of other antivirals. Adefovir is currently the treatment of choice in the event of resistance to lamivudine. Its effectiveness is confirmed by many studies and the risk of emergence of resistance is very low. Entecavir is a selective inhibitor of polymerase HBV and shows a better efficacy than lamivudine. It is well tolerated and is associated only with a weak risk of resistance, even after a prolonged treatment. PMID- 16212003 TI - [Wilson's disease: Clinical presentations]. AB - Wilson's disease is a rare genetic condition, transmitted on a recessive autosomal mode, which involves a disturbance of copper metabolism. Its prevalence is 1: 30000. It is treatable but may be lethal if not managed early and treated adequately. It is caused by the loss of function of an adenosine triphosphatase (ATP 7B), which is due to a mutation in the ATP 7B gene on chromosome 13. This leads to a decrease or absence of copper transport to the bile and its accumulation within certain organs, particularly the liver and the brain. In this article we present two cases of Wilson's disease in two young male patients. We also briefly review the pathophysiology of the illness, discuss the latest guidelines for diagnosis and treatment and outline the recent genetic discoveries. PMID- 16212004 TI - [Alcoholic and nonalcoholic steatohepatitis: the same disease! I. Diagnosis and mechanisms]. AB - Alcoholic steatohepatitis includes steatosis, inflammatory changes and hepatocellular damage. In severe form, jaundice and hepatic failure persist for several weeks, while non severe alcoholic steatohepatitis may follow an insidious course towards cirrhosis. Except for alcohol consumption, nonalcoholic steatohepatitis shares histological features and pathogenic mechanisms with alcoholic steatohepatitis, and is associated to the development of cirrhosis over time. Thus, given the increasing epidemics of the metabolic syndrome in industrialized countries, it is likely that alcoholic cirrhosis has been overdiagnosed in the past years. Obesity, insulin resistance and the oxidative stress including iron-mediated oxidative stress are involved both in alcoholic and non-alcoholic steatohepatitis. PMID- 16212005 TI - [Alcoholic and nonalcoholic steatohepatitis: the same disease! II. Management]. AB - Both alcoholic and non-alcoholic steatohepatitis share common histological findings and the risk of developing cirrhosis. Liver biopsy is required for the diagnosis, and the aim of treatment is to prevent cirrhosis. In alcoholic steatohepatitis, abstinence from alcohol is associated to prednisone in a severe form, as defined by a Maddrey's score > 32. Due to common pathogenic mechanisms, we recommend that coexisting metabolic alterations that are commonly observed in non-alcoholic steatohepatitis should also be treated in alcoholics. Thus, weight reduction, physical exercise, improvement of insulin sensitivity and reduction of oxidative stress, all may be beneficial both in alcoholic and non-alcoholic steatohepatitis. PMID- 16212006 TI - [Barrett's esophagus: epidemiology diagnosis and treatment in 2005]. AB - Barrett's oesophagus is a complication of gastro-oesophageal reflux disease and should be considered as a major risk factor for the development of adenocarcinoma of the oesophagus. Each and every patient with Barrett's oesophagus should be enrolled in a endoscopic surveillance program. During the last years, the endoscopic treatment of high grade dysplasia or intramucosal adenocarcinoma (by mucosectomy) has been reported to be successful in relatively large series of patients. This therapeutic option is particularly useful for patients with a high operative risk. It can be applied for lesions detected at an early stage only (i.e., usually detected in the setting of a surveillance program). Therefore, non operability is not considered as an exclusion criteria of such programs anymore. PMID- 16212007 TI - [Interventional analgesia justification in the era of evidence-based medicine]. PMID- 16212008 TI - [Treatment of congential heart disease with interventional catheterization]. AB - The development in pediatric interventional cardiology has changed the therapeutic strategy for many patients with congenital heart disease. Interventional catheterization became the first choice for valvuloplasty, angioplasty and collateral vessel occlusion. Effective and safe transcatheter interventions exist for closure of atrial or ventricular septal defects and for patent ductus arteriosus. The progress in the technology used in the catheterization laboratory will permit to continue the expansion of the range of interventions performed without surgery. Pediatric cardiologists and congenital heart surgeons must understand each other's interventional techniques and how they can be used in a coordinated fashion. This interaction is essential for the optimal management of patients with both simple and complex congenital heart disease. PMID- 16212009 TI - [Physician-patient relations and the capitalism of health care]. PMID- 16212010 TI - [Bird flu, this new menace that glides in our skies]. PMID- 16212011 TI - [Mad cow disease: towards a blood screening test]. PMID- 16212012 TI - [Patient advocacy]. PMID- 16212013 TI - Progress is not our most important product. PMID- 16212014 TI - Discrete mathematics in deaf education: a survey of teachers' knowledge and use. AB - The study documents what deaf education teachers know about discrete mathematics topics and determines if these topics are present in the mathematics curriculum. Survey data were collected from 290 mathematics teachers at center and public school programs serving a minimum of 120 students with hearing loss, grades K-8 or K-12, in the United States. Findings indicate that deaf education teachers are familiar with many discrete mathematics topics but do not include them in instruction because they consider the concepts too complicated for their students. Also, regardless of familiarity level, deaf education teachers are not familiar with discrete mathematics terminology; nor is their mathematics teaching structured to provide opportunities to apply the real-world-oriented activities used in discrete mathematics instruction. Findings emphasize the need for higher expectations of students with hearing loss, and for reform in mathematics curriculum and instruction within deaf education. PMID- 16212015 TI - Interviews with deaf children about their experiences using cochlear implants. AB - Within the framework of a longitudinal study of deaf children with cochlear implants, 11 children with implants were interviewed. The objective was to shed light on what it is like for a child to use a cochlear implant, based on these children's own experience with implants, which ranged from 5.0 to 7.5 years. Six of the children were in schools for the deaf, five in regular classes. All but one used an implant daily. The children appreciated that an implant enabled them to perceive sounds in the environment. Some of the children in regular classes could take part in one-to-one conversations with teachers but had difficulty following teaching and discussions. This observation was consistent with what the children's parents and teachers had maintained. Peer interaction was said to be best when other children had the use of at least some signs. PMID- 16212016 TI - Characteristics of the population of deaf and hard of hearing students with emotional disturbance in Illinois. AB - The study summarizes a database for the years 1994-1999 on deaf and hard of hearing students in Illinois with a diagnosis of emotional disturbance (N = 115). Data are reported on the group's demographic, domestic, etiologic, communication related, and intervention-related characteristics. These dually diagnosed students differed from Illinois's general population of deaf and hard of hearing students in many ways, including higher incidences of prematurity, prenatal trauma, and perinatal trauma. They were more likely to have had a later onset of hearing loss, to live in single-parent homes, to belong to an ethnic minority, to live in an urban or suburban area, and to qualify for low-income health care. Many had histories of abuse, 50% were regularly medicated, and 15% were assigned to surrogate parents. PMID- 16212017 TI - How physical education teacher education majors should be prepared to teach students with hearing loss: a national needs assessment. AB - A national needs assessment survey is described that gathered information on current practices in physical education in both center-based schools for the deaf and mainstream programs serving deaf and hard of hearing students, grades K-12. The manner in which deaf and hard of hearing students are being served in physical education programs, the depth and breadth of curricula, and the credentials needed to teach are described. The study compares similarities and differences among physical education programs in center-based deaf institutions and mainstream schools. In summary, the study identifies areas of concentration needed in curriculum, and methods of teaching appropriate for student teacher candidates. This information has value for physical education programs that are considering revising their curricula to prepare teacher candidates who wish to work with deaf students. PMID- 16212018 TI - Obstacles faced by deaf people in the criminal justice system. AB - Deaf people, especially those who are not well educated, are at risk for serious injustices when they enter the criminal justice system. The present study describes these risks at all stages of the legal process, including arrest, trial, probation, prison, and parole. These dangers are greatest for those who are poorly educated, read at a fourth-grade level or lower, have poor communication skills (American Sign Language and English), and lack awareness of their legal rights. Primitive personality disorder (PPD) is the term mental health professionals use to describe this set of characteristics. The risks that the segment of the deaf population with PPD faces when its members run afoul of the law are described, a case history provided, and some relevant legal and interpreting issues are discussed. A case is made for applying the concept of linguistic incompetence to deaf individuals with PPD. PMID- 16212019 TI - The effectiveness of international development assistance from American organizations to deaf communities in Jamaica. AB - American organizations bringing assistance to deaf people in developing countries unintentionally create relationships of dependency or oppression rather than relationships of support. Using qualitative methods, the author examined the effectiveness of development assistance provided to the Jamaican Deaf community by two American churches, one American nongovernmental organization, and one U.S. federal agency. Documents were reviewed and observations were made. Interviews were conducted with more than 60 deaf and hearing people involved with the American organizations, the Jamaican organizations, and deaf Jamaican beneficiaries. The author concludes that the Jamaican Deaf community was often excluded in planning, designing, or evaluating programs, and was unsatisfied with the American assistance it received. Results also indicate that the American organizations were poorly prepared to work with the Deaf community. Suggestions for American organizations wishing to strengthen and empower deaf people through development assistance in developing countries are proposed. PMID- 16212021 TI - ISMAs (involved safety meeting activities). PMID- 16212022 TI - Bored with keys? PMID- 16212020 TI - Reading comprehension and its relation to the quality of functional hearing: evidence from readers with different functional hearing abilities. AB - Three groups of students--19 hard of hearing, 20 deaf, and a control group of 36 typically developing hearing readers--were compared on their ability to process written words at the lexical level and on their comprehension of words within the structure of a sentence. Findings generally suggested that severe prelingual hearing loss does not prevent the development of word processing strategies adequate for efficient processing of written words at the lexical level, although such hearing loss seems to put individuals at risk of failure in internalizing syntactic knowledge crucial for proper processing of words at the sentence level. Evidence further indicated that neither the amount of functional hearing (deaf vs. hard of hearing), the hearing status of their parents (hearing impaired vs. hearing), nor the use of sign language as a primary communication mode was a direct cause in this regard. PMID- 16212023 TI - Facing reality. PMID- 16212025 TI - NFPA 70E: Performing the electrical flash hazard analysis. AB - Arc flash hazards pose a serious risk to employee safety; proper safe work practices must be utilized. Electrical equipment > or = 50 volts must be deenergized and locked out/tagged out prior to servicing and maintenance unless doing so would increase hazards or is infeasible. Remember, convenience is not an acceptable reason for keeping equipment energized during servicing and maintenance. If electrical equipment must remain energized during Servicing and maintenance, NFPA 70E should be consulted to determine flash hazard boundaries as well as required PPE. Finally, circuit breakers and electrical disconnects must be marked to warn qualified employees of potential arc flash hazards. PMID- 16212024 TI - Encouraging signs. PMID- 16212026 TI - Overcoming experience. PMID- 16212027 TI - Radiation detection. PMID- 16212028 TI - Confined space monitoring: is a PID sensor really necessary? PMID- 16212029 TI - Present and future opportunities. PMID- 16212030 TI - Mandibular third molar removal: patient preferences, assessments of oral surgeons and patient flows. AB - Mandibular third molar removal is one of the most common treatments conducted at oral and maxillofacial surgery clinics in Sweden. During the 1980's and 1990's, 20-25,000 mandibular third molars were removed annually which represents about 60% of the total operation volume. Removals performed in private specialist clinics and general dental clinics are not included in these figures. The aims of the present studies on mandibular third molars were to: 1) study values that reflect patients' preferences about possible outcomes of removal and non-removal; 2) make comparisons between Sweden and Wales with respect to patient's preferences; 3) study assessments of oral surgeons' indications for molars to be removed; 4) describe patient flows in the care process of removal. The multi attribute utility (MAU) method was used to quantify patients' preferences about outcomes following removal and non-removal. Whilst there were clear cultural and economic differences between the Swedish and the Welsh, there was a high degree of correlation in patients' ranking of the different outcomes for patients from the two countries (rs = 0.93, P<0.001). Generally, situations describing the outcomes of non-removal had a higher ranking than those describing the outcomes of removal i.e. patients seemed to prefer non-removal. Oral surgeons at seven specialist clinics registered data for 666 patients i.e. patient age and sex, the angular position and extent of eruption of the molar and whether or not there was an associated disease related to the molar proposed for removal. The indication for the removal was assessed on a Visual Analogue Scale (VAS), and the recorded results found to show a great variety. The mean VAS for removal of molars without disease was significantly lower than that for molars with associated disease. The differences between the mean VAS for molars with one disease compared with molars with two or three diseases were not significant. The patients' age was the only factor that had a significant effect on the assessment of the indication for molars without disease. The indication was higher for patients of the youngest age group than for patients of the oldest age group (P<0.05). In four specialist units in southern Sweden, the patient flows (the number of visits and what the visits comprise of) was registered for 361 patients. All details were recorded from arrival of the referral to the unit to performed mandibular third molar surgery. Eight different patient flows were found. The number of patient visits varied from one to three. For about 60 percent of the patients, attached radiographs to the referral were considered not appropriate and had to be completed, e.g. to be retaken. For a minority of the patients, the radiographic examination was completed at the radiological clinic included in the specialist unit and, in the oral and maxillofacial clinic for the others. The number of patient visits seemed not to depend on whether the attached radiographs were judged to be appropriate or not. IN CONCLUSION: Patient preferences seem to be more stable than the preferences of oral surgeons across the boundaries. Patients prefer outcomes of third molar non-removal as compared to outcomes following removal. Different patient flows may influence the cost-effectiveness in mandibular third molar surgery. PMID- 16212031 TI - [Calcium as a regulator of intracellular processes in actinomycetes: a review]. AB - Data on the effects of calcium ions (Ca2+) on processes of morphological and physiological differentiation in cultures of actinomycetes have been reviewed, with emphasis on representatives of the genus Streptomyces. Evidence accumulated thus far, of the regulatory role of serine-threonine protein kinases in the differentiation and of the possible involvement of Ca2+-dependent protein kinases in secondary metabolism (including antibiotic biosynthesis) are analyzed. The possibility that regulatory elements of apoptosis (including Ca2+-dependent) function in actinomycetes is discussed. A hypothesis is advanced, according to which determinants of antibiotic resistance play a key role in the network of signal transduction systems of actinomycetes. PMID- 16212032 TI - [Inhibition of peroxidase-catalyzed oxidation of chromogenic substrates by propyl gallate and its polydisulfide]. AB - Peroxidase-catalyzed oxidation of 2,2'-azino-di-(3-ethyl-2,3 dihydrobenzthiazoline-6-sulfonate) (ABTS) was competitively inhibited by propyl gallate (PG) and its polydisulfide (PGPDS) at 20 degrees C in 0.015 M phosphate citrate buffer (pH 6.0). Under these conditions, the values of the inhibition constant (Ki) were equal to 62 and 5.6 microM, respectively, for PG and PGPDS. The stoichiometric inhibition factor (f; the number of radicals extinguished per molecule of an inhibitor) equaled 2.0 and 14.7, respectively, for PG and PGPDS. Peroxidase-catalyzed oxidation of o-phenylenediamine was barely affected by PG or PGPDS. PGPDS may be used as a stop-reagent of peroxidase-catalyzed ABTS oxidation, whereas PG may serve as a calibrating inhibitor in test systems for measurement of total antioxidant activity (in human biological fluids, natural preparations, juices, wines, and other objects). PMID- 16212033 TI - [Polymerization of horseradish peroxidase in the presence of inorganic adsorbents]. AB - Efficiencies of binding between horseradish peroxidase (HRP) and its polymers (HRPp) with inorganic adsorbents (precipitated and coprecipitated) were studied. In aqueous solutions, HRP efficiently adsorbed to aluminum oxide and the coprecipitated sorbent (composed of calcium orthophosphate, magnesium hydroxide, and aluminum hydroxide). HRP readily bound to zinc hydroxide but not to aluminum hydroxide in 25.0 mM bicarbonate buffer (pH 9.0). Several variants of HRP polymerization and HRPp modification with diamines in the presence of Al2O3 and Zn(OH)2 were compared. Synthesis of HRPp according to the scheme comprising HRP activation in solution followed by its polymerization in the presence of Zn(OH)2 appeared the most efficient. HRP and HRPp bound to Zn(OH)2 displayed a high catalytic activity in the presence of high H2O2 concentrations. PMID- 16212034 TI - [Proteinase inhibitors as antistress proteins in higher plants]. AB - Physicochemical and functional characteristics of plant protein proteinase inhibitors as antistress biopolymers were studied to determine the mechanisms for plant resistance to phytopathogens and to obtain disease-resistant cereal and leguminous cultures. The activity of trypsin, chymotrypsin, and subtilisin inhibitors varied in monocotyledonous and dicotyledonous cultures. Study varieties of leguminous and cereal cultures were shown to contain endogenous inhibitors specific to proteinases of phytopathogenic fungi Fusarium, Colletotrichum, Helminthosporium, and Botrytis. These inhibitors were characterized by species specificity and variety specificity. Protease inhibitors from buckwheat seeds inhibited proteases of fungal pathogens and suppressed germination of spores and growth of the fungal mycelium. Our results suggest that proteinaceous inhibitors of proteinases are involved in the protective reaction of plants under stress conditions. PMID- 16212036 TI - [Metabolites of marine organisms as regulators of O-glycosylhydrolases]. AB - The ability of metabolites contained in culture liquid of 62 strains of marine fungi to affect the activity of two digestive enzymes of marine mollusks--endo 1,3-beta-D-glucanase of Spisula sachalinensis and beta-D-glucosidase of Littorina kurila--was studied. It was found that 66 and 71% of specimens activated, 18 and 7% inhibited, and 16 and 22% did not affect the activity of endo-1,3-beta-D glucanase and beta-D-glucosidase, respectively. It is demonstrated that the metabolites of brown algae and marine sponges can be used for a targeted regulation of enzyme biosynthesis by marine fungi. The protein inhibitor of endo 1,3-beta-D-glucanases isolated from the brown alga Laminaria cichorioides blocked the biosynthesis of almost all O-glycosylhydrolases in five strains of marine fungi studied. The presence in culture medium of halistanol sulfate from the marine sponge of the family Halichondriidae either did not affect or activated the biosynthesis of enzymes involved in carbohydrate metabolism by marine fungi. PMID- 16212035 TI - Purification of bovine milk lactoperoxidase and investigation of antibacterial properties at different thiocyanate mediated. AB - Bovine lactoperoxidase (LPO) was purified with amberlite CG 50 H+ resin, CM sephadex C-50 ion-exchange chromatography, and sephadex G-100 gel filtration chromatography from skim milk. The activity of lactoperoxidase was measured by using 2.2-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) diammonium salt (ABTS) as a choromogenic substrate at pH 6.0. Purification degree for the purified enzyme was controlled with SDS-PAGE and Rz value (A412/A280). Rz value for the purified LPO was 0.8. Km value at pH 6.0 at 20 degrees C for the LPO was 0.20 mM. Vmax value was 7.87 micromol/ml min at pH 6.0 at 20 degrees C. Bovine LPO showed high antibacterial activity in 100 mM thiocyanate--100 mM H2O2 medium for some pathogenic bacteria, such as Aeromonas hydrophila ATCC 7966, Micrococcus luteus LA 2971, Mycobacterium smegmatis RUT, Bacillus subtilis IMG 22, Pseudomonas pyocyanea, Bacillus subtilis var. niger ATCC 10, Pseudomonas aeruginosa ATCC 27853, Enterococcus faecalis ATCC 15753, Bacillus brevis FMC3, Klebsiella pneumoniae FMC 5, Corynebacterium xerosis UC 9165, Bacillus cereus EU, Bacillus megaterium NRS, Yersinia enterocolytica, Listeria monocytogenes scoot A, Bacillus megaterium EU, Bacillus megaterium DSM32, Klebsiella oxytocica, Staphylococcus aerogenes, Streptococcus faecalis, Mycobacterium smegmatis CCM 2067 and compared with well known antibacterial substances such as penicilline, ampicilline, amoxicillin-clavulanate and ceftriaxon. The LPO--100 mM thiocyanate- 100 mM H2O2 system was purposed as an effective agent against many of the diseases causing organisms in human and animals. PMID- 16212037 TI - [Optimizing the conditions of dextran synthesis by the bacterium Leuconostoc mesenteroides grown in a molasses-containing medium]. AB - Maximal dextran production (54-55 g/l) by the bacterium Leuconostoc mesenteroides strain V-2317D was observed in molasses-containing media in the presence of 17.5% glucose at pHinit 6.75. The beginning of dextran production depended on the amount of inoculate; maximum yield was observed at a shaker rate of 200 rpm. The dextran produced by L. mesenteroides grown in the molasses-containing medium was representative of three fractions differing in the molecular weight and composition: the high- (approximately 54.5%), medium- (approximately 27.9%), and low-molecular-weight (approximately 2.85%) fractions. PMID- 16212038 TI - [Structural characteristics and biological properties of Pseudomonas fluorescens lipopolysaccharides]. AB - The structure and biological properties of lipopolysaccharides (LPSs) from strains IMB 4125 (=ATCC 13525) and IMB 7769 of the bacterium Pseudomonas fluorescens (biovar I) were studied in vitro. LPSs were similar in the composition of lipid A and the core lipid but differed in the structure of O specific polysaccharide chains, which was corroborated by the absence of serological relationships between them. The toxicity (LD50) of LPSs of P. fluorescens with respect to D-glucosamine-sensitized mice was 40-50 times lower than the toxicity of the classic endotoxins, LPSs of E. coli. The LPSs studied stimulated the production of tumor necrosis factor (TNF) and nitric oxide (NO) by mouse peritoneal macrophages. The rates of TNF and NO synthesis induced by the LPSs of interest were eight to nine and three to five times lower, respectively, than the corresponding parameters of the control LPSs of E. coli 055:B5 and 026:B6. Additionally, LPS preparations of the P. fluorescens strains induced TNF synthesis by monocytes of human whole-blood preparations. Certain differences in biological properties of these strains have been revealed, which could be due to the characteristic features of LPS structure and composition in different cultures. PMID- 16212040 TI - [Determination of minimal concentrations of biocorrosion inhibitors by a bioluminescence method in relation to bacteria, participating in biocorrosion]. AB - By using a bioluminescence ATP assay, we have determined the minimal concentrations of some biocorrosion inhibitors (Katon, Khazar, VFIKS-82, Nitro-1, Kaspii-2, and Kaspii-4) suppressing most common microbial corrosion agents: Desulfovibrio desulfuricans, Desulfovibrio vulgaris, Pseudomonas putida, Pseudomonas fluorescens, and Acidithiobacillus ferrooxidans. The cell titers determined by the bioluminescence method, including not only dividing cells but also their dormant living counterparts, are two- to sixfold greater than the values determined microbiologically. It is shown that the bioluminescence method can be applied to determination of cell titers in samples of oil-field waters in the presence of iron ions (up to 260 mM) and iron sulfide (to 186 mg/l) and in the absence or presence of biocidal corrosion inhibitors. PMID- 16212039 TI - [Anaerobic microbial associations degrading aromatic amino acids]. AB - Anaerobic microbial associations have been isolated that degrade aromatic amino acids to methane and carbon dioxide at high rates. Significant differences between the morphological, cytological, and physiological traits of cultures isolated from samples of adapted and unadapted sludge are shown. The effects of cultivation temperature, illumination, and presence of mineral nitrogen and bicarbonate in the medium upon adaptation of enrichment cultures to substrates and subsequent behavior of the anaerobic associations have been studied. Intermediate and final products of degradation of aminoaromatic compounds and the sequence of their formation in the cultures have been determined. We have also studied the effects of exogenous electron acceptors and additional carbon sources on the degradation of aminoaromatic compounds. PMID- 16212041 TI - [Monitoring of microbial degraders in manned space stations]. AB - Samples of microorganisms from the surface of constructions of Mir Space Station (Mir SS) were taken and examined after 13 years of operation. The following microorganisms were isolated and identified: 12 fungal species belonging to the genera Penicillium, Aspergillus, Cladosporium, and Aureobasidium; 3 yeast species belonging to the genera Debaryomyces, Candida, and Rhodotorula; and 4 bacterial species belonging to the genera Bacillus, Myxococcus, and Rhodococcus. The predominant species in all samples was Penicillium chrisogenum. It was shown that the fungi isolated could damage polymers and induce corrosion of aluminum magnesium alloys. We commenced a study of microbial degraders on constructions of the Russian section of the International Space Station (RS ISS). Twenty-six species of fungi, bacteria, yeasts, and actinomycetes, known as active biodegraders, were identified in three sample sets taken at intervals. We founded a collection of microorganisms surviving throughout space flights. This collection can be used to test spacecraft production materials, in order to determine their resistance to biodegradation. PMID- 16212042 TI - [Extracellular proteolytic enzymes of Azospirillum brasilensis strain Sp7 and regulation of their activity by a homologous lectin]. AB - It was found that Azospirillum brasilensis strain Sp7 is able to produce extracellular proteolytic enzymes. The enzymes were active within a broad range of pH values, with two peaks of activity being located in the acid and alkaline pH areas; required calcium ions; and exhibited substrate specificity with respect to azogelatin. Zymography allowed at least four proteolytic enzymes with molecular weights of 32, 45, 52, and 174 kDa to be detected in A. brasilense Sp7 culture liquid. It was shown that the lectin from A. brasilense Sp7 can inhibit proteolytic enzymes. PMID- 16212043 TI - [Lipid composition of cells of heterothallic strains in the developmental cycle of Blakeslea trispora]. AB - Lipid compositions in mycelium and spores of Blakeslea trispora heterothallic strains were studied. Distinctions between the strains in the ability to synthesize linolenic acid and in optimal growth temperature were demonstrated. The (-) strain grew at a higher temperature and was unable to synthesize linolenic acid, whereas the (+) strain accumulated this acid up to 20% of total fatty acids. The distinctions between the strains remained at different developmental stages (mycelium and spores). A higher thermophilicity of the (-) strain correlated with a high sterol content, which is typical of thermophilic fungi. The lipid compositions of heterothallic strains studied differed in lipid content, their fractional composition, the degree of unsaturation, and carotenoid composition. PMID- 16212044 TI - [Immobilized yeast membranes as biocatalysts for sucrose inversion]. AB - Yeast membranes were obtained by autolysis of various strains with relatively high invertase activity. Heterogeneous biocatalysts for sucrose inversion were made of the yeast membranes and granulated carbon-containing supports made of common natural materials: expanded clay aggregate (ECA), sapropel, and lignin. The properties of these biocatalysts were studied. It was shown that the biocatalyst activity and stability of the immobilized yeast membranes increased with reference to the initial ECA, independent of the structure of the carbon layer synthesized on the support surface. Heterogeneous biocatalysts prepared by adsorption of yeast membranes on sapropel had the greatest activity and stability, whereas lignin-based biocatalysts were relatively unstable. PMID- 16212045 TI - [Sterols of mulberry leaves and small leaf curl disease]. AB - Free and bound sterols of leaves of five mulberry cultivars differing in their susceptibility to small leaf curl disease have been studied. The total content of sterols in all samples is similar and is not correlated with the resistance of the cultivars. The qualitative composition of particular sterols is also identical. They are represented by cholesterol, campesterol, stigmasterol, sitosterol, and two 4alpha-methylsterols. The leaves of the most sensitive cultivar are characterized by high cholesterol content. The ratio sitosterol : stigmasterol decreased in proportion to the resistance level of a cultivar. PMID- 16212046 TI - [Sorption of components from a mixture of essential oils by cryotextured cornstarches]. AB - Sorption by cryotextured cornstarches of components of the aqueous phase of a mixture of essential oils was studied by capillary gas chromatography. The amount of cryotexture-sorbed substances depended linearly on their concentration in the initial gel. The sorption of components from the mixture by starch polysaccharides was mainly associated with hydrophobic cooperative interactions, which resulted in the formation of supramolecular structures and inclusion complexes. The structure of the compounds was a major factor determining the degree of sorption. Sorption of monoterpene carbohydrates was the most pronounced. We revealed a synergistic increase in the degree of sorption from the mixture as compared to binding of individual compounds. PMID- 16212047 TI - [Molecular logic: complementarity, learning, and comparison of images in molecular world]. AB - It was shown that, using intramolecular transformations and based on the complementarity principle, it is possible to carry out the learning of a molecular system (memorizing of a complex input influence) and subsequent multiple comparison of a new signal with this signal. PMID- 16212048 TI - [Low-reactive free radicals inhibiting nonbranched chain processes of addition]. AB - The kinetics of inhibition of nonbranched chain processes of addition of reactive free radicals at double bonds O=O, C=O, and C=C of molecules in liquid homogeneous binar systems from saturated and unsaturated components by low reactive free radicals (o-CH3C6H4CH2O*4, HO*4; HC=O; CH2=C(CH3)CH2, and CH2=CHCHOH) was studied. It was assumed that as the concentration of the unsaturated component (as a source of low-reactive free radicals) increases (after the optimum concentration is attained, which corresponds to the maximum rate of the process) the phenomena of increasing inhibition of these processes may act as the elements of self-regulation of similar processes in nature, which returns them into the stationary state. The energetics of the key radical molecular gas phase reactions was analyzed. The thermal effects of total dissociation reactions of simple alkylperoxyl (exo effect) and alkoxyl (endo effect) reactive free radicals in the gas phase were compared. The kinetic equations (with one to three parameters being directly determined), obtained by the quasi-steady-state treatment, are presented, which make it possible to describe nonmonotonic (with a maximum) dependencies of the rates of formation of molecular 1:1-adducts on the concentration of the unsaturated component. PMID- 16212049 TI - [The superoxide-scavenging activity of some amino acids in water solutions]. AB - The superoxide-scavenging activity of a number of amino acids was revealed. The activity is comparable in effectiveness with known antioxidants. An original method for the estimation of antioxidant properties of amino acids was proposed. Quantum mechanical calculation of the interaction of lysine with superoxide anion was made. It was assumed that the mechanisms of the biological action of some physical factors, such as weak magnetic fields adjusted to the cyclotron frequency of ions of some amino acids, are associated with the modulation of the antioxidant activity of these molecules. PMID- 16212050 TI - [The peculiarities of fluorescence of natural steroid hormones, excited by a laser source of the UV range]. AB - The data of studies were summarized that demonstrated for the first time the fluorescence of the members of all classes of steroid hormones. The fluorescence spectra were characterized, the relative fluorescence quantum yields were estimated, and the dependence of the intensity and spectrum of fluorescence on the presence and position of side groups and double bonds in the molecule of the hormone was established. PMID- 16212051 TI - [Structure-function organization of neurokinin A and neurokinin B molecules. III. A conformational study of glycine-monosubstituted analogues of neurokinins A and B]. AB - The conformational features of some glycine-monosubstituted analogues of neurokinins A and B were investigated by the method of theoretical conformational analysis. The calculated geometry and energy parameters permitted one to determine the structural role of each of these substituted amino acids in the mechanism of folding of the low-energy conformational states of neuropeptides. On the basis of the calculated data and the results of biological tests of these analogues, the structure-function relationships of neurokinins A and B were discussed. PMID- 16212052 TI - [The component analysis of tryptophan fluorescence spectra of melittin during its oligomerization]. AB - The oligomerization of melittin with increasing ionic strength and protein concentration was investigated using the methods of decomposition of its tryptophan fluorescence spectra into "elementary" log-normal components. At high ionic strength (up to 2 M KCl), the emission spectra of tetrameric melittin are well described as the sum of two log-normal components, suggesting the presence of tryptophan residues in two sorts of environment with greatly differing polarity. Measurements of fluorescence spectra by iodide showed that these two spectral components possess different Stern-Volmer constants, that is, the tryptophans emitting them have different solvent accessibility, which does not correlate with the crystallographic structure of tetrameric melittin. Moreover, in the oligomerization transition induced by ionic strength, the tetrameric intermediate is formed, which has log-normal spectral components with relative contributions differing from those in 2 M KCl. PMID- 16212053 TI - [The mechanism of oxymyoglobin oxidation catalyzed by ferrocyanide ions: chemically modified and mutant sperm whale myoglobins]. AB - A comparative study of the rate of ferrocyanide-catalyzed oxidation of native sperm whale MbO2, its chemically modified derivative in which all accessible His residues are alkylated by sodium bromoacetate, (CM-MbO2), and mutant sperm whale MbO2 with His119 replaced by Asp residiue, [MbO2(His119-->Asp)] was carried out. The influence of pH, ionic strength, and [Fe(CN)6]4- concentration on the oxidation rate was investigated, as well as the effect of complexing MbO2 with redox-inactive Zn2+ ion, which, at the equimolar Zn2+ concentration, forms a stable complex with His119(GH1) on the protein surface. It was shown that the mechanism of the catalysis involves specific binding of [Fe(CN)6]4- to the protein at the His119(GH1) region, which is in agreement with a large positive electrostatic potential and the presence at this site of Mb of a cavity large enough to accommodate [Fe(CN)6]4- anion. The protonation of nearby His113 and His116 residiues (especially of the latter) plays a very important role in the catalysis, promoting the fast oxidation of bound [Fe(CN)6]4- by dissolved oxygen. Only the presence of these both necessary conditions in MbO2 structure provides its effective oxydation catalyzed by ferrocyanide. PMID- 16212055 TI - [The properties of sequences involved in insertion and deletion mutagenesis in rodents]. AB - We analyzed nucleotide sequences that were inserted to or deleted from genomic DNA during the divergence from common ancestor. The median length of these sequences is 3 nucleotides; they are enriched with palindromes and often repeat in adjacent DNA. PMID- 16212054 TI - [A study of interactions of carbonic anhydrase B with water and urea. I. Spin diffusion parameters that determine individual and cooperative properties of molecules]. AB - The rigidity parameter (G), which is characteristic of protein compactness, was studied in native globular carbonic anhydrase B. The dependence of parameter G on power and excitation time of spin-diffusion was expressed analytically. We found out that native carbonic anhydrase B is able to form water-protein units that are probabilistically distributed with respect to their sizes. Large water-protein units can be detected by analyzing the spin-diffusion spectra. The excitation frequencies of spin-diffusion spectra were shifted far away from typical 1H NMR spectra of carbonic anhydrase B. PMID- 16212056 TI - [A comparative study of the lipid phase in native and circulating multiple modified low-density lipoproteins of human blood by the use of the spin probe method]. AB - Different approaches based on the spin probe method were used to compare the physical state of the surface lipid monolayer in subfractions of low-density lipoproteins: in native low-density lipoproteins constituting the bulk of human blood low-density lipoproteins and in circulating multiple-modified low-density lipoproteins whose portion is minor in healthy persons but significantly increases in atherosclerotic patients. The data obtained in in vitro experiments suggest that circulating multiple-modified low-density lipoproteins possess atherogenic properties. The order parameter S, rotational correlation time tau, and hydrophobicity parameter h were calculated from electron spin resonance spectra of a series of spin probes whose paramagnetic groups are located at different depths of the lipid monolayer. These parameters characterize the molecular packing, fluidity, and polarity in the microenvironment of paramagnetic groups. The kinetics of the reduction of paramagnetic groups by ascorbate and oxidation by hypochlorite were obtained for the spin probe whose paramagnetic group is located deeply in the lipid monolayer at the level of the terminal segments of phospholipid acyl chains. No difference between native low-density lipoproteins and circulating multiple-modified low-density lipoproteins was revealed in respect of the physical properties of the lipid domain of surface proteolipid layer, as sampled by spin probes. PMID- 16212057 TI - [A correlation between respiration and synthesis of ATP in mitochondria at different degree of uncoupling of oxidative phosphorylation]. AB - It is known that mitochondrial respiration in state 3 is due to three simultaneous and independent processes: synthesis of ATP (1), endogenous passive proton leakage (2), and proton leakage by protonophoric uncoupler (3). The total rate of processes (2) and (3) is equal to the product of respiration rate in state 4 and coefficient KR, which is defined as the ratio of the deltamuH+ value in state 3 to that in state 4. It is shown that it is possible to calculate both the rates of processes (1), (2) and (3) separately and the protonophoric activity of uncoupler using the coefficient KR and other coefficients, which are determined as the ratio of deltamuH+ values in state 3 or in state 4 to its maximal value. Simple methods of determination of these coefficients were developed, which are based on the study of the dependence of respiration rate in states 3 and 4 on the concentration of protonophoric uncoupler. It was found that the uncoupling action of palmitate, a natural uncoupler of oxidative phosphorylation, unlike classic uncoupler-protonophores DNP and FCCP, depends not only on its protonophoric activity but also on the inhibition of the process (1). PMID- 16212058 TI - [High-order derivative spectroscopy of infrared absorption spectra of the reaction centers from Rhodobacter sphaeroides]. AB - The infrared absorption spectra of reduced and chemically oxidized reaction center preparations from the purple bacterium Rhodobacter sphaeroides were investigated by means of high-order derivative spectroscopy. The model Gaussian band with a maximum at 810 nm and a half-band of 15 nm found in the absorption spectrum of the reduced reaction center preparation is eliminated after the oxidation of photoactive bacteriochlorophyll dimer (P). This band was related to the absorption of the P(+)y excitonic band of P. On the basis of experimental results, it was concluded that the bleaching of the P(+)y absorption band at 810 nm in the oxidized reaction center preparations gives the main contribution to the blue shift of the 800 nm absorption band of Rb. sphaeroides reaction centers. PMID- 16212059 TI - [The influence of new hybrid antioxidants ichphans on the kinetics of ascorbate induced reduction of spin probe radical centers in liposomes]. AB - The influence of new hybrid antioxidants ichphans on the structure of liposome lipid bilayer was studied. The analysis of the kinetics of the ascorbate-induced reduction of spin probe radical centers incorporated in a membrane revealed that ichphans produce a modifying effect on the structure of liposome membrane. It was established that derivates with distinct hydrophobic properties transform different sites of the membrane that are the sites of localization of ichphans in the intramembraneous space. The data obtained suggest that lipid components play an important role in the effects of ichphans on biological membranes. PMID- 16212060 TI - [Membrane proton conductivity and energy-dependent fluxes of hydrogen ions in bacteria Enterococcus hirae grown in media with different pH values]. AB - It was shown that the proton conductivity of Enterococcus hirae ATCC9790 membrane increases three times as pH of the growth medium decreases from 8.5 to 5.5. The changes in proton conductivity are interrelated to the values of membrane and redox potentials of the cell, which in turn vary depending on the pH value of growth medium. The energy-dependent H+ efflux for cells fermenting sugar (the glucose) decreases 1.5 times as pH decreases from 8.5 to 5.5; in this case, the N,N'-dicyclohexylcarbodiimide at lower pH values suppresses the H+ efflux more intensively than at higher pH values, the H+ efflux nonsensitive to N,N' dicyclohexylcarbodiimide being practically unchanged. The H+ efflux in the ATPase mutant MS116 is significantly (approximately 3 times) lower than that in the precursor strain and does not depend on pH. The results show that the proton conductivity of the membrane of this bacterium depends on pH of the growth medium. It is possible that the energy-dependent H+ efflux through F1F0-ATPase is interrelated with membrane proton conductivity. PMID- 16212061 TI - [The stochastic resonance of magnetosomes fixed in the cytoskeleton]. AB - The rotation of microscopic magnetic particles, magnetosomes, embedded into the cytoskeleton and subjected to a magnetic field and thermal noise was considered. The dynamics of magnetosome is shown to comply with the conditions of the stochastic resonance under not too tight constraints on the character of particle fastening. The excursion of regular rotations attains the value of the order of radian, which facilitates explaining the biological effects of low-frequency weak magnetic fields and geomagnetic fluctuations. PMID- 16212062 TI - [Indirect and repeated electromagnetic irradiation of extremely high freguency of bacteria Escherichia coli]. AB - It has been shown that separate irradiation of distilled water and tris-phosphate buffer containing some inorganic ions, with Escherichia coli K12 grown in anaerobic conditions upon fermentation of sugar (glucose) with "noise" electromagnetic radiation of extremely high frequencies (53.5-68 gHz) or millimeter waves (wavelength of 3 to 8 mm) with low flux capacity (0.01 mW) for 10, 30 and 60 min caused opposite effects, changing the growth of these bacteria. The irradiation of water has a bactericide effect, whereas the irradiation of the buffer stimulates bacterial growth although the buffer itself inhibits the growth. These results point out the role of water in the bactericide action of "noise" electromagnetic radiation of extremely high frequencies, and confirm the significance of membranotropic effects. The bactericide action disappeared after repeated irradiation for 10 and 30 min with 2-h intervals. This indicates the operation of some compensatory mechanisms in bacteria. PMID- 16212063 TI - [On the mechanism of the nonspecific cell response to the action of damaging agents and the nature of hormesis]. AB - A conception of the molecular mechanism of the universal nonspecific cell response (NCR) to the action of all damaging factors of physical and chemical nature is described. The mechanism is shown to be closely related to the general principles of the structural-functional organization of cells and to the properties of biological macromolecules. Among them are the phenomenon of nonspecific regulation of the enzyme activity by low-molecular substances and the compartmentation of the latter within the cell, which is provided by the system of intracellular membranes. Special attention is paid to the role of disturbance of the barrier function, of membranes in the development of the nonspecific cell response, and the activation of repair of cell injuries. Based on the theory of nonspecific cell response, the stimulating effect of weak action of agents that damage the cells at high doses (the phenomenon named as hormesis) is explained. PMID- 16212064 TI - [Synchronization of mechanochemical auto-oscillations within the Physarum polycephalum plasmodium by periodical external actions]. AB - Amoeboid locomotion of huge unicellular organism, the Physarum polycephalum plasmodium, is stipulated by endoplasmic flow, which is produced by spatially highly coordinated rhythmic contractions of the ectoplasm. To describe the self organization of the plasmodial contractile activity, we proposed a mathematical model, which is based on the hypothesis of positive feedback between the deformation of the cytoskeleton and release of a chemical regulator of the active contraction. A nonautonomous analogue of this model was used to study the synchronization of mechanochemical auto-oscillations by periodic gradient of the external pressure. Numerical computations of the system of differential equations obtained revealed a dependency of the synchronization band on the amplitude of the external pressure oscillations. On the basis of this dependence and experimental data on the band of synchronization of the shuttle endoplasmic flow by the periodic gradient of temperature obtained with the help of the laser Doppler anemometer, relative efficiency of external synchronizing action of temperature and pressure was evaluated. PMID- 16212066 TI - [Theoretical aspects of evaluation of the plasma membrane time constant in myocardial tissue using a model of three-dimensional RC-medium of finite thickness]. AB - An analytical solution of the differential equation describing the distribution of the electrotonic potential in the three-dimensional RC-medium of finite thickness when the current source is hemispheric was obtained. Computerized programs were developed, and families of curves of T1/2 (time during which the electrotonic potential reaches half of the stationary amplitude) dependence on the size of current electrode (R0), the distance between the current electrode and the potential recording site (R), and medium thickness (D) were established. It is shown that function T1/2 = f(R) is not linear in close vicinity of the current electrode, and the application of linear dependence T1/2 = R/2 + const for the determination of the plasmic membrane time constant (taum) in myocardial tissue would lead to errors of -70% / +20%. PMID- 16212065 TI - [The role of blood plasma porphyrins in the effects of He-Ne-laser on human leukocytes]. AB - The He-Ne-laser induced effects in human blood leukocytes in the presence of autologic plasma were investigated. Experiments were performed in two ways: (1) He-Ne-laser irradiation of cells in the presence of autologic plasma or (2) laser irradiation and subsequent addition of autologic plasma to the cell suspension. The concentration dependencies of plasma additions were evaluated. To obtain different concentrations of porphyrins in plasma samples, we either diluted the samples with PBS or selected patients with different porphyrin plasma content. The effects of He-Ne-laser irradiation were characterized by the maximum effect dose (Dmax) of irradiation and the degree of maximum cell activation (Amax, priming index). In the first series of experiments, we irradiated leukocytes in the presence of autologic plasma taken from patients with pneumonia and bronchial asthma. It was found that Dmax decreased with increasing porphyrin concentration in plasma. It was observed that, at low porphyrin concentrations, Amax increased severalfold with increasing photosensitizer concentration. At a porphyrin concentration of 0.46 pmol a decrease in Amax was detected as the porhyrin concentration increased. The same effects were revealed at high doses of laser irradiation. Very similar effects were found in experiments with the addition of irradiated plasma to cells. However, the Amax value was considerably less compared to that after irradiation in the presence of plasma (160% vs. 230 - 270% upon combined irradiation of cells and plasma). The Dmax value was higher in the series of experiments in which plasma was irradiated separately. The results suggest that laser-induced leukocyte activation can be mediated by blood plasma porphyrins and the products of lipid peroxidation formed as a result of porphyrin photosensitized lipid oxidation. PMID- 16212067 TI - [Shaping processes in drying drops of serum in norm and pathology]. AB - The results of physical and mathematical simulation of the drying process for colloid liquids under different initial conditions were considered. It was shown that the concentration and stability of a colloid solution play an important role in the shaping of drying drops. Possible mechanisms of the formation of morphological differences in dried drops of serum in healthy and ill people are discussed. PMID- 16212068 TI - [The mechanism of verbal memory and scenario of the development of speech V rhythms]. AB - It was shown that the Feigenbaum scenario for the transition of bifurcation of period doubling to chaos, which explains the singularities of V-rhythm disorders in the neighborhood of the critical point, is a good model of the development of phonetics in children speech. It was also shown that the singularities of the dynamics of V-rhythms in the bifurcation lacuna in the zone of chaos intrinsic to the Pomeau-Manneville scenario for route of bifurcations of period 3 to chaos are capable, basically, to describe some features of both remembrance at the extraction from memory during speech and remembering in memory during perception of speech. PMID- 16212069 TI - [A new mathematical model of the dynamics of an age cohort population]. AB - A new generalized conception of an organism is given. Based on this conception, a new mathematical model of ontogenesis of an individual and the survival of the age cohort of population was proposed. By using real data on the dynamics of the survival of the age cohort of population, the model enables one to determine the parameters characterizing the relationship man-environment in the context of survival and calculate the dynamics (from birth to death) of the model variables of the state of the organism. PMID- 16212070 TI - [The effect of biota on global climate]. AB - A model consisting of two blocks (equations) was proposed for the analytical study of the biosphere-climate system over great periods of time. The first equation describes the balance of carbon dioxide in the atmosphere and represents the biological block of the model. The second equation is the equation of the energy balance or the physical block of the system. The model is based on the most general conceptions of living matter and the evolution process. A possible interpretation of some events and phenomena in the earth history in terms of the model is given. PMID- 16212071 TI - [The apology of bioinformatics]. AB - The discussion about adequate understanding of the term "bioinformatics" is continued. The relationships between bioinformatics and experimental molecular biology are considered. The list of the main branches and achievements of modern bioinformatics is presented. PMID- 16212072 TI - Current perspective. Diabetic vascular disease: from endothelial dysfunction to atherosclerosis. AB - Cardiovascular complications are the leading cause of death in diabetes. Over the past decade a number of studies have addressed the underlying mechanisms. Derangements of endothelial function, also referred to as endothelial dysfunction, have emerged to be the crucial early step in the development of atherosclerosis and are also involved in plaque progression and clinical emergence. Endothelial dysfunction is a condition of impaired endothelium dependent vasodilation and most important of "endothelial activation", characterized by a proinflammatory, proliferative, and procoagulatory milieu that promotes initiation and complications of atherogenesis. A synergistic cross-talk among the conventional cardiovascular risk factors associated with diabetes contributes to disruption of endothelial integrity and accelerated atherosclerosis. This review will focus on the multifactorial nature of endothelial dysfunction in diabetes, the relationship between endothelial dysfunction, conventional cardiovascular risk factors and atherosclerosis, and the therapeutic options to improve endothelial function. PMID- 16212073 TI - Prognostic role of non-sustained ventricular tachycardia in a large cohort of patients with idiopathic dilated cardiomyopathy. AB - BACKGROUND: The identification of patients with idiopathic dilated cardiomyopathy (IDC) at higher risk of sudden death (SD) is still an unsolved issue, and the role of non-sustained ventricular tachycardia (NSVT) uncertain. METHODS: The effect of NSVT on total mortality, SD and life-threatening arrhythmias was evaluated in 554 patients with IDC on optimal medical treatment and at long-term follow-up (81 +/- 58 months). RESULTS: At diagnosis, 240 patients (43%) had NSVT at Holter monitoring and 314 (57%) did not. During follow-up, 189 patients (5/100 patients-year) died or underwent heart transplantation; SD occurred in 53 patients (1.4/100 patients-year); SD + non-fatal ventricular arrhythmias occurred in 75 patients (2/100 patients-year). Patients with and without NSVT at diagnosis had the same 5-year transplant-free survival rate (76 vs 76%, p = NS) and a similar incidence of SD (10 vs 7%, p = NS). The length and rate of NSVT did not show any significant relationship with the outcome. Only heart failure symptoms (NYHA class III-IV) (hazard ratio [HR] 1.9, p = 0.015) and severe left ventricular impairment (left ventricular ejection fraction < or = 0.30 and left ventricular end-diastolic diameter > or = 70 mm) (HR 2.7, p < 0.0001) were independently associated with higher SD risk. At multivariate analysis the presence of frequent NSVT episodes (> or = 3 runs/day) was associated with an increased risk of total mortality (HR 1.68, p = 0.041) and of major ventricular arrhythmias (HR 2.11, p = 0.037), but only in the subgroup of patients with severe left ventricular impairment. CONCLUSIONS: Patients with advanced heart failure symptoms, severe left ventricular dysfunction and dilation had a higher risk of SD independently of NSVT. The finding of more frequent NSVT was associated with an increased risk of all-cause mortality and of major ventricular arrhythmias in patients with severe left ventricular impairment. PMID- 16212074 TI - DDD versus VVIR versus VVI mode in patients with indication to dual-chamber stimulation: a prospective, randomized, controlled, single-blind study. AB - BACKGROUND: The aim of this study was to compare VVI, VVIR and DDD modes in patients with indication to dual-chamber stimulation, depending on left ventricular function. METHODS: Two groups of patients were implanted with a DDD pacemaker: Group I with ejection fraction > 40% and Group II with ejection fraction < 40%. Patients with a history of atrial arrhythmia or retrograde conduction were excluded. At follow-up (1 month each) quality of life (QoL), patient preference and echo parameters were collected. At hospital discharge all patients were programmed in DDD for 1 month and then randomized to VVI or VVIR mode. At the end of the period in VVI or VVIR mode each patient underwent a control period in DDD and then was programmed in VVIR or VVI mode. RESULTS: Seventeen patients out of 23 preferred DDD mode and 6 did not perceive any subjective difference among DDD, VVI and VVIR modes (4/9 in Group I and 2/14 in Group II, p = 0.0017). QoL was significantly different between the two groups and at each follow-up showed the best values in DDD. The correlation between QoL and Tei index was 0.62 in Group I (p < 0.001) and 0.35 in Group II (p = 0.001). Neither ejection fraction nor fractional shortening showed any significant difference during the three phases of the study. CONCLUSIONS: Most patients preferred the DDD mode. The Tei index showed a good correlation with QoL and both QoL and Tei index significantly improved with DDD mode as compared to VVI and VVIR. PMID- 16212075 TI - Predictive parameters of left ventricular reverse remodeling in response to cardiac resynchronization therapy in patients with severe congestive heart failure. AB - BACKGROUND: Cardiac resynchronization therapy (CRT) is useful for the treatment of severe congestive heart failure. Unfortunately up to 30% of patients could be non-responders. The aim of our study was to find parameters to predict responsiveness to CRT. METHODS: Fifteen patients (9 males, 6 females, mean age 67.3 +/- 7.8 years, range 52-83 years) with dilated cardiomyopathy, NYHA functional class III-IV, left ventricular (LV) ejection fraction < 35% and QRS > or = 110 ms, underwent CRT. All the patients had echocardiographic evidence of systolic dys-synchrony. RESULTS: One patient died of electromechanical dissociation. The remaining 14 patients maintained biventricular stimulation at 6 months; mean QRS width decreased from 156 to 132 ms (p < 0.001). Ten patients (71%) were considered responders because of a reduction in LV end-systolic volume > 15%. In non-responders (4 patients, 29%) LV end-systolic volume was stable in 3 patients and increased in 1. LV ejection fraction significantly increased only in responders (p < 0.001). Responders had more severe pre-pacing dyssynchrony than non-responders (p < 0.001). Inter- (p = 0.002) and intraventricular dyssynchrony (p = 0.003) did significantly reduce after CRT only in responders. On multiple regression analysis there were two independent predictors of reverse remodeling after pacing: the baseline mitral QS-tricuspid QS (QSm-QSt) time (B = -1.7, p = 0.005) and the intraventricular dyssynchrony index (B = -1.55, p = 0.007). Pre implant QSm-QSt of 38 ms correctly identified the two groups: responders had a value > 38 ms and non-responders < 38 ms. The pre-implant intraventricular dyssynchrony index of 28 ms was the cut-off value: responders had an index > 28 ms, non-responders < 28 ms. CONCLUSIONS: In the literature a tissue Doppler imaging index of intraventricular dyssynchrony evaluated before implantation is used to select responders to CRT. In our work we studied interventricular and intraventricular dyssynchrony, and both the QSm-QSt time and the standard deviation of the 12 LV segment QS time were correctly able to identify responders. PMID- 16212076 TI - Off-pump versus on-pump coronary artery bypass: does number of grafts performed represent a selection bias in comparative studies? Results from a matched cohort comparison. AB - BACKGROUND: Several retrospective studies comparing off-pump and on-pump coronary surgery and the largest randomized studies published to date showed a lower number of grafts performed in patients submitted to off-pump coronary artery bypass surgery (OPCAB). These findings bring about the question of the general applicability of the results. We eliminated the selection bias correlated with the number of grafts per patient by comparing the short-term outcomes of patients undergoing OPCAB and standard coronary artery bypass grafting (CABG) matched for number of grafts. METHODS: Eighty-seven consecutive patients undergoing OPCAB (group A) were selected from the database of our Institution during a 2-year period. Matching was performed by iterative selection prioritizing, in the following sequence: number of grafts, EuroSCORE, and age. A total of 87 patients operated upon with the on-pump technique represented the control group (group B). RESULTS: There were no significant differences in preoperative characteristics between the two groups. The number of grafts per patient was 2.2 +/- 0.5 in group A and 2.2 +/- 0.5 in group B. Early mortality did not differ between the two groups and it was 2.2% (2 patients) in group A and 3.4% (3 patients) in group B (p = NS). The incidence of myocardial infarction did not differ between the two groups. No patient in either group had stroke or coma. Five (5.7%) patients in group A and 7 (8.0%) patients in group B had atrial fibrillation (p = NS). CONCLUSIONS: We were unable to demonstrate any significant differences in short term mortality or morbidity outcome between OPCAB and standard CABG patients Our findings suggest that excellent results can be obtained with both surgical approaches. PMID- 16212077 TI - Impact of pulmonary regurgitation and age at surgical repair on textural and functional right ventricular myocardial properties in patients with tetralogy of Fallot. AB - BACKGROUND: The aim of this study was to identify non-invasively the potential impact of pulmonary regurgitation and age at surgical repair on the right ventricular (RV) textural and functional myocardial properties in patients operated on for tetralogy of Fallot (TOF). METHODS: We assessed the average intensity (Int.(1B)) and the cyclic variation (CV(IB)) of the echocardiographic backscatter curve in 30 TOF patients (mean age 16.2 +/- 8.3 years), who had undergone corrective surgery (mean age at repair 3.2 +/- 2.6 years, range 0.2-11 years). They were divided into three age- and body surface area (BSA)-matched subgroups according to the results of the surgical repair: 12 patients had no significant postsurgical sequelae (group I), 12 patients had isolated moderate severe pulmonary regurgitation (group II), and 6 patients had pulmonary regurgitation associated with significant (> 30 mmHg) RV outflow tract obstruction (group III). In addition, 30 age-, sex- and BSA-matched normal subjects were identified as the control group. RESULTS: In our study population, CV(IB) was lower (7.86 +/- 2.5 vs 10.6 +/- 1.4 dB, p < 0.001) and Int.IB higher ( 18.6 +/- 4.1 vs -21 +/- 2.8 dB, p = 0.01) compared to the control group. Comparison between the control group and each subgroup of TOF patients showed: a) comparable values of CV(IB) and Int.(IB) in group I (10.6 +/- 1.4 vs 9.4 +/- 2.3 dB, p = 0.07; and -21 +/- 2.8 vs -21.4 +/- 2.3 dB, p = 0.7, respectively); b) Int.(IB) was significantly different only in group III (-21 +/- 2.8 vs -13.3 +/- 4.6 dB, p < 0.0001), c) CV(IB) was different either in group II or III (10.6 +/- 1.4 vs 7.42 +/- 2, p < 0.001; and 10.6 +/- 1.4 vs 5.56 +/- 1.8, p < 0.001, respectively). In addition, comparison of integrated backscatter indexes among the TOF subgroups revealed significant differences of CV(IB) between group I and II (9.4 +/- 2.4 vs 7.4 +/- 2, p = 0.03) and between group I and III (9.4 +/- 2.4 vs 5.56 +/- 1.8, p = 0.004), and of Int.(IB) between group I and III (-21.4 +/- 2.3 vs -13.3 +/- 4.66, p < 0.001) and between group II and III (-21.4 +/- 2.3 vs 18.6 +/- 2.8, p = 0.006). Group III patients, who had the most significant RV dilation, expressed as the ratio between RV and left ventricular end-diastolic diameter (0.55 +/- 0.8) compared to group II (0.67 +/- 0.11, p = 0.038) and group I (0.55 +/- 0.87, p < 0.001), showed the lowest values of CV(IB) (5.56 +/- 1.8 dB) and the highest values of Int.(IB) (-13.3 +/- 4.6 dB) Finally, in our study population, both the degree of RV dilation and the age at surgical repair significantly correlated with Int.(IB) (r = 0.49 and r = 0.4, p = 0.06 and p = 0.033, respectively) and inversely correlated with CV(IB) (r = -0.55 and r = 0.53, p = 0.002 and p = 0.003, respectively). CONCLUSIONS: In patients operated on for TOF: a) integrated backscatter analysis may identify patients with significant RV myocardial abnormalities related to postsurgical sequelae; b) residual pulmonary regurgitation, particularly if associated with pulmonary stenosis, appears to affect RV myocardial properties; c) an earlier repair of TOF may result in better preservation of myocardial characteristics. PMID- 16212078 TI - Ventricular capture management in pediatric pacing: efficacy and safety. AB - BACKGROUND: The Ventricular Capture Management (VCM) of Medtronic Kappa 700 series pacemakers (PM) performs automatic threshold detection and optimization of pacing output that may enhance generator longevity. We evaluated efficacy and safety of this algorithm in children. METHODS: The study was prospective, non randomized, involving 50 consecutive patients (mean age 5.6 +/- 6.6 years, median 4 years), enrolled at first PM implant. VCM was active from the implant, with nominal values of safety margin, minimum adapted pulse amplitude and width. Leads were endocardial and epicardial, all unipolar. Thresholds and pacing outputs were registered with telemetric PM interrogation. Endocardial and epicardial thresholds and outputs were also compared. Follow-up duration was 27 +/- 13 months (range 6-49 months). RESULTS: A significant reduction in pulse amplitude was evident since the sixth month. Thresholds and outputs were lower in endocardial than in epicardial pacing. A false negative capture detection occurred during the "acute phase" in 3 patients (6.0%), with incorrect automatic output increase to 5 V/1 ms. After this phase, the problem was still detected in 2 patients (4.0%). VCM correctly identified threshold increases in 2 patients (1%). No pacing defect was documented. VCM was not performed in 4 infants (8.0%) for pacing rate > or = 100 b/min. CONCLUSIONS: VCM function is safe and effective in reducing pacing output in pediatric patients; this may increase PM longevity. Epicardial pacing shows higher thresholds and outputs than endocardial pacing. PMID- 16212079 TI - Initial energy for biphasic external electrical cardioversion of atrial fibrillation. AB - BACKGROUND: No international guidelines indicate the initial energy in biphasic external electrical cardioversion of atrial fibrillation (AF) actually. The aim of this study was to determine this value in order to find a reasonable compromise between the necessity of limiting tissue damage and of quickly restoring sinus rhythm as well. METHODS: Fifty-six consecutive patients with AF candidate to external electrical cardioversion were treated using adhesive anterior-posterior paddles and biphasic wave defibrillator Lifepack 12, with steps of 50 J. After 6 hours troponin I levels were measured. RESULTS: Thirty four patients were cardioverted by 50 J (group A), 18 by 100 J (group B) and 3 by 150 J (group C). One patient was not cardioverted (success rate 98%). No significant differences were noted between groups A and B with regard to age, sex, weight, height, thoracic circumference, body mass index, body surface area, impedance, NYHA class, left ventricular ejection fraction, left atrial diameter, causes of heart disease, antiarrhythmic medications, and duration of current AF episode. No increase of troponin I levels occurred. CONCLUSIONS: An initial shock of 100 J in the biphasic external elective cardioversion of AF is a valid and highly effective option. An initial shock of 50 J was effective in 61% of our population, and it is probably appropriated in patients with a lower weight and body mass index. PMID- 16212080 TI - Brugada syndrome and neurally mediated susceptibility. AB - The risk of sudden death in patients with Brugada syndrome (BS) is still unclear. Moreover, particular clinical conditions may have a confounding effect on the diagnostic and therapeutic approach. We report the case of a 27-year-old man with a clinical history of suspected neurally mediated syncope and typical ECG features of BS. The tilt table test showed a type I, mixed, positive response. The electrophysiological study (EPS) disclosed a peculiar ventricular irritability with the induction of a life-threatening arrhythmia. After the implantation of a cardioverter-defibrillator an episode of ventricular fibrillation during sleep at night was correctly identified and treated by the device. The association between neurally mediated susceptibility and the typical ECG abnormalities of BS is not an unexpected event in young subjects. The misjudgment of the pathophysiological mechanism of syncopal episodes may lead, on one hand, to overlook the risk of sudden death and, on the other, to pursue inappropriate therapeutic measures. The application of a tailored diagnostic work up based on currently available guidelines may be useful to overcome the clinical and therapeutic dilemma. PMID- 16212081 TI - Two unusual cases of coincident atrioventricular nodal reentrant tachycardia and ventricular tachycardia. AB - The simultaneous occurrence of narrow and wide QRS complex tachycardias was observed in 2 patients evaluated at our electrophysiological centers. Electrophysiological testing revealed the coexistence of two types of arrhythmia (atrioventricular nodal reentrant tachycardia and verapamil-sensitive left ventricular tachycardia) in one patient and of three types of arrhythmia (atrioventricular nodal reentrant tachycardia, ventricular tachycardia originating from the right ventricular outflow tract, and left ventricular tachycardia) in the other. Both patients underwent successful radiofrequency ablation of all the types of tachycardia. PMID- 16212082 TI - A difficult diagnosis: right unilateral cardiogenic pulmonary edema. Usefulness of biochemical markers of heart failure for the correct diagnosis. AB - We describe a case of unilateral pulmonary edema occurring in a young woman affected by hypertrophic cardiomyopathy complicated by acute worsening of mitral regurgitation. The relevant role of biochemical markers of heart failure, such as brain natriuretic peptide and carbohydrate antigen 125, in clarifying the final diagnosis of cardiogenic pulmonary edema and modifying treatment accordingly is emphasized. PMID- 16212083 TI - Percutaneous treatment of the left main coronary artery ostial obstruction following aortic valve replacement. AB - Iatrogenic left main coronary artery ostial stenosis is a rare and late life threatening complication of aortic valve replacement. The exact causes of this critical condition, despite being still nowadays elusive, are possibly related to the insertion of perfusion catheters into the left coronary system for cardioplegia delivery. We describe the case of a 69-year-old man, with normal coronary arteries documented by preoperative coronary angiography before surgery, who developed 1 year after aortic valve replacement worsening effort angina. A second coronary angiography revealed a severe left main ostial stem stenosis, which was successfully treated by sirolimus-eluting stent deployment. This case demonstrates a new percutaneous approach of this poorly understood, yet potentially fatal complication following aortic valve replacement. PMID- 16212084 TI - Congestive heart failure secondary to right ventricular metastasis of colon cancer. A case report and review of the literature. AB - Although metastatic tumors of the heart occur more frequently than primary ones, infiltration of the right heart by a metastatic colon cancer has rarely been reported. We report the case of a woman previously operated on for colon cancer, presenting with symptoms of congestive heart failure due to metastatic invasion of the right ventricular cavity. Both transthoracic and transesophageal echocardiography were useful in detecting the mass, but not in defining its nature. The patient underwent a palliative surgical resection of the neoplastic mass but died soon after the intervention. PMID- 16212085 TI - [Detection of Aedes (Stegomyia) Aegypti L. mosquitoes in Sochi city]. AB - Few Aedes aegypti females were found when collecting the mosquitoes attacking human beings in the Central District of Sochi in August to September 2001-2004. Ae. aegypti, a vector of dangerous causative agents of diseases, such as yellow and Aden fevers, appeared on the Black Sea coast of the Caucasus is recorded after its long absence. By taking into account the potential epidemic value of Ae. aegypti, it is necessary to make a monitoring in the cities, towns, and settlements to establish the spread, number, and the breading sites of mosquitoes in the given area and to prevent their mass reproduction. The effectiveness of Ae. albopictus as a vector of Aden fever has been established in different regions of the world. Entomological surveys for Ae. albopictus should be made in the areas of Russia where Ae. aegypti mosquitoes were distributed early in the past century, particularly in the southern port towns and settlements of Russia. Ae. albopictus is potentially able to spread to the north further than is Ae. aegypti. PMID- 16212086 TI - [RAPD analysis of the populations of the malaria mosquitoes Anopheles superpictus and A. pulcherrimus in the malaria foci of the central Asia]. AB - RAPD-analysis was used to estimate intraspecies-specific variability in malaria mosquitoes, the major vectors of malaria in Central Asia. The high heterogeneity of RAPD-loci was found in 7 populations of An. superpictus and 2 populations of An. pulcherrimus. Genetic distances between the populations were determined on the basis of analysis of the frequencies of 15 RAPD-loci. There was a genetic similarity of An. superpictus populations in the malaria foci of Tadjikistan with those in the neighboring Surkhandariya Region of Uzbekistan. PMID- 16212087 TI - [Distribution and control of acari as vectors of human disease in urban population of Moscow]. AB - Human ectoparasites, such as itch mites and the follicular mites Demodex brevis and D. folliculorum are notified on the territory of Moscow. The bird and rodent mites Ornythonyssus bacoti and the larvae of harvest mites may attack human beings indoors. The ticks Ixodes ricinus and I. persulcatus begin to inhabit the tree belts and environs of Moscow and other cities and towns. In the cities and towns of more southern regions of the Russian Federation, the red hen-mites Dermanyssus gallinae are also encountered and the ticks Rhipicephalus sanguineus and Ornithodoros papillipes begin to inhabit urban structures. A differential system of their controlling measures is proposed in relation to the habitats of mites and ticks on the urban territories and the results of use of a number of currently available disinfectants are presented. PMID- 16212088 TI - [Efficiency of control of Ixodes ticks, vectors of natural and focal infections in the south of the Tyumen Region]. AB - There are synchronous undulating variations in the number of ticks and in the incidence of tick-borne encephalitis and Ixodes tick-borne borreliosis in the south of the Tyumen Region. Since 1990s, there has been an increase in the incidence of tick-borne infections and in the number of ticks. The highest epidemic risk areas of 1418 have been treated with acaricides due to the additional financing made by the Tyumen Region in 2002-2004. The treatments with the agents Sipaz-super and Briz (25% c.e. cipermethrin) have ensured 98-100% freedom of the areas from ticks. In Tyumen and in the south of the Tyumen Region, the number of human suction of ticks has reduced from 601 and 1231 per 100,000 in 2001 to 424 and 658 cases in 2004, respectively. The authors have developed guidelines for tick-controlling measures in large areas of the Tyumen Region. PMID- 16212090 TI - [Comparative assessment of the diagnostic value of the laboratory diagnostic methods for trichomoniasis]. AB - The authors compared the sensitivity and specificity of currently available methods for laboratory diagnosis of trichomoniasis, by examining 971 persons. The highest frequency of T. vaginalis was detected by studies of a stained smear (37.4%), culture tests (19.0%); polymerase chain reaction (PCR) (17.1%), immunofluorescence tests (12.7%), wet smear test (2.7%). Enzyme immunoassay yielded positive results in 36.2% of cases. The use of PCR and culture tests frequently provided similar results (92.0%). The authors have proposed an algorithm of a laboratory study including wet smear microscopy, PCR, and culture tests as basic methods. They have noted the higher detection rates of T. vaginalis when analyzing during draining physiotherapeutic procedures (endocervical vibratory massage with vacuum aspiration in females and pneumovibratory massage of the prostate with endourethral chemotrypsin electrophoresis in males). PMID- 16212089 TI - [Organization and tactics of the ground tick control in the south of the Tyumen Region]. AB - Measures for protection of the general population from the attacks of vectors of natural and focal infections were implemented by OOO "Spetsbioservis" at the expense of the regional budget in the involving foci oftick-borne encephalitis (TBE) and Ixodes tick-borne encephalitis in the south of the Tyumen Region during 2002-2004. The organizational and tactical issues of ground antitick treatments were perfected during this work. The paper describes the maximum effective completeness of teams of disinfectors and their supply with materials and equipment, as applied to local conditions. This scheme of organizational and tactical measures for acaricidal treatments in the south of the Tyumen Region in 2002-2004 has indicated their high effectiveness and mobility. In 2002-2003, tick bites were not recorded in children's health facilities. In these years, the incidence of TBE in the population of Tyumen reduced by 39.3% as compared with 2001. The qualitative implementation of acaricidal treatments, their extended scopes, successively intensified sanitary educational work among the population, and introduced personal prophylactic agents (repellents and protective clothes) will substantially affect the epidemiological situation associated with tick borne infections in the region. PMID- 16212091 TI - [Toxoplasma gondii antibodies in domestic animals in the town of Vologda]. PMID- 16212092 TI - [Role of toxoplasmas in pathology of the vision organ]. AB - The present paper presents data on the major clinical symptoms ofToxoplasma oculopathy in different forms of Toxoplasma infection. It also gives the results of the authors' own laboratory studies of specimens taken from individuals with pathology of the vision organ. They confirm the data available in the literature on that toxoplasmosis frequently occurs in combination with other infections, particularly with Herpes and Cytomegalovirus, Chlamydia, Ureaplasma, and Mycoplasma infections. The signs of eye lesions and a clinical case of an eleven year-old girl diagnosed as having central recurrent multifocal exudative and hemorrhagic neurochoriorenitis of the right eye are given. PMID- 16212093 TI - [The specific features of development of Echinococcus granulosus metacestode (exogenous blastogenesis) in man]. AB - The paper describes a case of abnormal development of a parasitic cyst by the type characteristic of alveolar echinococcosis (exogenous blastogenesis) detected in a female patient operated on for echinococcus. The fact that the exogenous blastogenesis has been found allows the mechanism of a postoperative recurrence of human echinococcosis to be explained from basically new standpoints. A parasitological analysis of the punctates of cystic masses and cysts themselves, taken from patients operated on for cystic echinococcosis has revealed one of the signs of malignant exogenous proliferation of elements of a parasytic metacestode, namely, the development of acephalocysts from the germinative cells of the pedicle of protoscoleces. The capacity of cystic echinococcosis metacestodes for exogenous blastogenesis by the type of human alveolar echinococcosis is an early unknown species-specific feature of E. granulosus. PMID- 16212094 TI - [Opisthorchiasis in the Republic of Mordovia in 2000 to 2003]. PMID- 16212095 TI - [Prevalence of toxocariasis in rural and urban areas]. AB - The prevalence of toxocariasis was studied among different age groups of the population in eight municipal entities of the Sverdlovsk Region. Since 1995, the number of infected patients was shown to have annually increased by 1.4-2 times and amounted to 2.9 per 100,000. The 2003 serological surveys indicated that Toxocara antibodies were found in 8.9% of the children examined in different areas of the region. The epidemic process more actively involved rural children. Numerous tests for Toxocara eggs were more positive in the cities and towns with every amenity than in rural areas. Serodiagnostic tests demonstrated that children aged 3-6 and 7-14 years were involved into the epidemic process. The similar data were obtained in different age groups of adults. Nevertheless, among the adult population, the number of adult seropositive persons was significantly 1.9 times higher than that of children, which may reflect certain regularities in the formation of an immune stratum. The titers of serum Toxocara antibodies, being 1:800 or higher, may suggest that more than 50% of the total number of the examinees might be, in terms of clinical signs, patients with toxocariasis. PMID- 16212096 TI - [Gastrointestinal function in patients with psoriasis concurrent with chronic opisthorchiasis before and after antiparasitic therapy]. AB - The impact of antihelminthic therapy on gastrointestinal function was studied in 150 patients with psoriasis concurrent with chronic opisthorchiasis, 100 patients with psoriasis without helminthiasis, and 30 healthy individuals. A fractional study of gastric secretion was conducted in its both phases, by using histamine stimulation; carbohydrate absorption was investigated by the 5-g D-xylose test; fat absorption by the Kamer procedure; the bioelectrical activity of the small bowel was examined by electric enterography. These indices were followed up for 2 3 years. There were negative changes in gastric function in patients with 2-year persistent Opisthorchis invasion as compared with those recovered from this condition, in the latter, the pathological process becoming stable. After 3 years of the follow-up, negative changes were found in the values of fat and D-xylose absorption in patients with psoriasis as compared to the cured patients in whom the indices did not significantly become worse, However, in the cured patients, the absorption of fat and Dxylose did not normalize. The 3-year follow-up indicated that the patients with psoriasis concurrent with chronic opisthorchiasis had significantly decreased amplitudes of propulsive and non propulsive waves (p < 0.05 and p < 0.01, respectively) while the latter indices were significantly increased in the cured patients (p < 0.01 and p < 0.05, respectively), but failing to reach the normal values. Effective dehelminthization in patients with psoriasis concurrent with chronic opisthorchiasis improved gastrointestinal function, making the course ofpsoriasis better (there were no exacerbations for 2 years). PMID- 16212097 TI - [Use of microwave ovens in the prevention of opisthorchiasis]. AB - The studies were performed to evaluate the impact of microwave radiation on the larval stages of parasites and particularly on the survival of Opisthorchis metacercariae when fish was cooked in domestic microwave ovens. It was found to take less time to free fish and its products from Opisthorchis metacercariae in the microwave oven than that to cook the product completely. The performed experiments have shown that the use of microwave ovens to domestically prepare carps from unfavorable opisthorchiasis water reservoirs is an effective mean of freeing fishes from Opisthorchis metacercariae. PMID- 16212098 TI - [Intestinal parasitic diseases in HIV-infected patients in Uzbekistan]. AB - Intestinal parasitic diseases were diagnosed in 100 HIV-infected patients at different stages of disease (its asymptomatic form, persistent generalized lymphoadenopathy, pre-AIDS, and AIDS) (Group 1), 100 Tashkent residents (Group 2), and 349 patients with gastrointestinal diseases, allergic dermatoses, and skin depigmentation foci (Group 3). The HIV-infected patients were found to have virtually all parasites, such as Giardia lamblia, Cryptosporidium parvum, Chilomastix mesnili, Entamoeba coli, Iodamoeba butschlii, Entamoeba histolytica/dispar, Endolimax nana, Blastocystis hominis, Enlerobius vermicularis, Ascaris lumbricoides, Hymenolepis nana, detectable in the population of Tashkent. The highest infestation with intestinal protozoa, including nonpathogenic amoebas and helmninths, was found in Groups 1 and 3. However, in all the forms of HIV infection, the infestation with E. histolytical/dispar was 10 times greater than that in Groups 2 and 3 (1% and 0.8%, respectively). G. lamblia was detected in 16, 21, and 45.2% in Groups 1, 2, and 3, respectively. In all the HIV-infected patients, the content of CD8 lymphocytes was increased, but that of CD20 lymphocytes was normal. Parasites were detectable with different levels of CD4 lymphocytes, but C. parvum was found only if its count was > 200/ml. In the HIV-infected patients, the hyperproduction of IgE was caused mainly by helminths rather than protozoa. In these patients, the increased level of IgE was also noted in the absence of parasites. PMID- 16212099 TI - [Acaricidal activity of "Medifox" against the human itch mites Sarcoptes scabiei]. AB - The acaricidal activity of Medifox was evaluated in terms of the itch mites to lend support to the validity of its regimen to treat human scabies. Experiments using mainly females, eggs, and larvae withdrawn from the burrows on the skin of patients with scabies were performed. Permethrin contained in 0.4% aqueous Medifox emulsion was found to affect an itch mite by contact and intestinal routes, without a marked knock-down effect; however, the signs of intoxication are irreversible and lead to the death of Medifox-treated mites. In an in vitro experiment, 100% death of specimens occurred 24-48 hours after immersion of the active stages in the aqueous emulsion for 1 min, depending on the stage of their development. Visual inspection of the course of embryogenesis and the hatch of larvae from eggs under a light microscope established that 20-40-min exposure to Medifox emulsion did not prevent the final formation of a larva and its hatch if the egg was at the late stages of embryogenesis. Long-term (180-min) exposure yielded a 100% ovicidal effect. An in vivo experiment indicated that there were no viable mites in the scabies burrows after single-to-double treatment of patients. Repeated applications are required due to their incomplete ovicidal effect. As a matter of fact, the third treatment is additional to create the "safety factor" of therapy. It is concluded that there is an agreement of the results of in vitro and in vivo experiments, by studying Medifox as an example. The findings suggest that the chosen Medifox treatment regimen including three consecutive treatments results in complete eradication of all the stages of the causative agent on the patient. PMID- 16212100 TI - [External respiratory function in echinococcosis]. AB - The paper presents the results of a study of the parameters of external respiratory function in 33 patients with echinococcosis. Restrictive respiratory dysfunction was detected in 85.71% of the examinees. After inhalation tests, better parameters of external respiratory function were documented. PMID- 16212101 TI - Effects of tamoxifen dose and nutrition scheme during growth on stimulation of the reproductive system in cornish breed cocks. AB - The effects of tamoxifen (TAM) antiestrogen dose as well as protein and energy content in feed on stimulation of the reproductive system development in meat type cocks were studied. The experiment was conducted on males of the Dominant White Cornish chicken breed. The optimum TAM dose for stimulation of reproductive system development was at the level of 5 mg per kg body weight. The effects of TAM administration on the following traits were investigated: body weight, breast muscle thickness, age at sexual maturity, egg fertilization and hatchability. No effect of TAM on breast muscle thickness was found. It was also noted that semen obtained from 11 to 12 week old males of the Dominant White Cornish breed could be used successfully in the insemination of females. PMID- 16212102 TI - Effects of Bacillus thuringiensis kurstaki on Malpighian tubule cells of Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae) larvae. AB - In this study effects of Bacillus thuringiensis kurstaki (Btk) on Malpighian tubule cells of Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae) larvae was investigated by electron microscopy. 3 mg/l Btk was given with food. After Btk administration, the Malpighian tubule cells were investigated and compared with a control group. 3 and 6 hrs after Btk administration swelling in Malpighian tubule cells was observed. Swelling of mitochondria and separation of their cristae was seen after 12 hrs. After 24 hrs dissolution of the basal cytoplasm, swelling and vacuolization of all mitochondria, partial dissolution of the nucleoplasm, and swelling and separation ofmicrovilli was documented. A membrane-body in the nucleus was seen after 48 hrs. The nucleoplasm was completely dissolved after 72 hrs and after 96 hrs large vacuoles appeared in the cytoplasm and shortening of microvilli was observed. PMID- 16212103 TI - Assessment of the effect of alpha-galactosides injected during embryogenesis on selected chicken traits. AB - The effect of different doses of alpha-galactoside (RFOs) preparations from Pisum sativum L. cv. Opal, injected into eggs during embryogenesis, on maintaining a high number of bifidobacteria, selected chicken broiler traits and the lipoprotein level of blood were studied. Two independent experiments were conducted. In the first, Ringer water solution containing 1.763 mg/egg of fructooligosaccharides (FOS) (I group), 2.1158 mg of pea RFO preparation containing 20% sucrose (II group) and 0.4232 mg of sucrose (III group) were injected into Hubbard broiler breeder eggs containing 12-day old embryos. Only Ringer water solution was applied to the eggs of the control group (IV group). The number of bifidobacteria determined in faeces of two-day old chicken of groups I and II was significantly higher in comparison with the sucrose and control groups. The high level ofbifidobacteria of groups I and II was maintained during 6 weeks. The dose of both preparations had no influence on the body weight, carcass, breast muscle, leg and abdominal fat ratio, total cholesterol, HDL and LDL serum concentrations. Broiler mortality and breast muscle cholesterol concentration was highest (P < 0.05) for the control group. On the other hand, the European Production Index, as well as serum triglycerides, were the lowest for this group. The second experiment was performed on Hybro G chicken breeder eggs. 0.69, 3.43 and 6.87 mg/egg of pea RFO preparation doses containing 20% sucrose were injected into the experimental groups. The number of bifidobacteria in the caecum and selected meat traits of broilers were determined. The results of this experiment confirmed that RFO injection in ovo causes the long-time maintenance of a high level ofbifidobacteria. The dose of the preparations does not have any effect on the selected broiler meat traits, except that the highest dose increases the percent of carcase in body weight. However, this dose reduced the hatchability of the treated embryos. PMID- 16212104 TI - Comparison of proliferation and motile activity between human keratinocytes isolated from skin and oral mucosa. AB - OBJECTIVE: Epithelial wound repair assures the recovery of the epithelial barrier after wounding. During wound healing epithelial cells migrate to cover the wound surface. For healing of skin wounds the skin keratinocytes can be replaced by oral mucosa epithelial cells grown in vitro. The presented experiments were carried out in order to compare the proliferation, morphology, and migration between human keratinocytes isolated from human skin and oral mucosa. MATERIALS AND METHODS: Human epidermal and oral mucosa keratinocytes from primary culture were used in all experiments. Cell motility and shape were determined using computer-aided methods. RESULTS AND CONCLUSIONS: It was demonstrated that although both cell types exhibit the same typical epithelial morphology, oral mucosa keratinocytes locomote significantly faster than skin keratinocytes. They also differ in proliferation activity. Oral mucosa keratinocytes exhibited faster growth and different actin cytoskeleton organisation than skin keratinocytes under in vitro conditions. Autologous oral mucosa keratinocytes may be expanded in vitro and used for skin wound healing in vivo. PMID- 16212105 TI - Myotomal myogenesis of axial muscle in the sturgeon Acipenser baeri (Chondrostei, Acipenseriformes). AB - Compared to teleost fishes, a unique character of the myogenesis of the plesiomorphic A. baeri is the fusion of myoblasts derived from the somite, leading to the formation of multinucleate muscle lamellae. Then, the lamellae are converted into cylindrical muscle fibres. The mechanism of transformation of lamellae into fibres is still debatable. Early embryonic muscle growth is mainly due to the hypertrophy of somite-cell derived stock. After hatching, hypertrophic growth occurs parallel to hyperplastic growth. Proliferatively active mesenchymal cells, which migrate from the intermyotomal space into the myotomes, participate in both processes. PMID- 16212106 TI - The origin of syncytial muscle fibres in the myotomes of Xenopus laevis--a revision. AB - During the early stages of myogenesis in X. laevis, the primary myoblasts (of mesodermal origin) differentiate simultaneously, in each myotome, into mononucleate myotubes. At later stages mesenchymal cells appear in intermyotomal fissures and then in the myotomes between myotubes and contribute to the formation ofsyncytial muscle fibres. The pathway of mesenchymals cell during myogenesis was described in X laevis by monitoring the incorporation of 3H thymidine. 3H-thymidine was incorporated in the nuclei of mesenchymal cells in intermyotomal fissures of younger myotomes and then in those of older myotomes between the myotubes revealing the proliferation of mesenchymal cells. As expected, nuclei of differentiating mononucleate myotubes did not incorporate 3H thymidine. At later stages of myogenesis the myotubes were found to contain two classes of nuclei: large nuclei of the primary myoblasts (of myotomal origin) and smaller nuclei originating from secondary myoblasts ofmesenchymal origin. TEM and autoradiographic analyses confirm that mulinucleate myotubes in X. laevis arise through fusion of secondary myoblasts with mononucleate myotubes. PMID- 16212107 TI - Ultrastructural studies of the ovary of Palaeococcus fuscipennis (Burmeister) (Insecta, Hemiptera, Coccinea: Monophlebidae). AB - Ovaries of Palaeocoocus fuscipennis are composed of about 100 telotrophic ovarioles that are devoid of terminal filaments. In the ovariole a tropharium ( = trophic chamber) and vitellarium can be distinguished. The tropharium contains 7 trophocytes. A single oocyte develops in the vitellarium. The oocyte is surrounded by follicular cells that do not undergo diversification into subpopulations. The obtained results are discussed in a phylogenetic context. PMID- 16212108 TI - Structure of the ovary and the differentiation of follicular epithelium in the pig louse, Haematopinus suis (Insecta: Phthiraptera). AB - The female reproductive system of the pig louse, Haematopinus suis (Insecta: Phthiraptera) is composed of paired ovaries, lateral oviducts, and a common oviduct that leads into a vagina. Clusters of mycetocytes (= cells filled with symbiotic organisms) are associated with lateral oviducts. Each ovary is composed of five loosely arranged ovarioles of the polytrophic-meroistic type. An individual ovariole is covered by a basal lamina and is composed of a terminal filament, germarium, and vitellarium. The terminal filament is composed of large, disc-shaped cells that are orientated perpendicularly to the long axis ofthe ovariole. The basal part of the terminal filament is separated from the germarium by a well-developed transverse septum. The germarium is short and filled with clusters of oogonial cells. In each cluster the cells arejoined by intercellular bridges, filled with fusomal material. Within the cluster, only one cell, the future oocyte, enters the prophase of the first meiotic division; the other cells differentiate into nurse cells. The basal part ofthe germarium is filled with the somatic prefollicular cells. The boundary between the germarium and the vitellarium is not distinct. The vitellarium contains linearly arranged ovarian follicles in subsequent stages of oogenesis (previtellogenesis, vitellogenesis and choriogenesis). Each follicle consists of an oocyte and 7 nurse cells and is surrounded by follicular cells. During oogenesis the follicular cells diversify, so that ultimately, five morphologically distinct subpopulations of these cells can be distinguished: (1) cells in contact with the nurse cells, (2) anterior cells, (3) mainbody cells, (4) posterior cells, and (5) interfollicular cells. Interestingly, the follicular cells associated with the anterior part of the oocyte, i.e. located in space at the oocyte/nurse cell border (fold cells) are mitotically active throughout previtellogenesis. It might be suggested, in this context, that the separation of the oocyte from the nurse cell compartment is brought about by mitotic divisions, consequent multiplication and centripetal migration of these cells. PMID- 16212109 TI - Recent data on the occurrence of species of the Paramecium aurelia complex in Europe. AB - Among 15 species of the Paramecium aurelia complex known world-wide, 10 have been found in Europe, namely: P. primaurelia, P. biaurelia, P. triaurelia, P. tetraurelia, P. pentaurelia, P. sexaurelia, P. septaurelia, P. novaurelia, P. dodecaurelia, and P. tredecaurelia. Recent data on the frequency of occurrence of the species in Europe are given in the paper. PMID- 16212110 TI - Evidence of B-chromosomes in the karyotype of Barypeithes pellucidus Boheman 1834 (Coleoptera, Curculionidae, Entiminae) from Central Europe. AB - B-chromosomes were observed in spermatogonial mitotic metaphases, meiotic metaphases I and II of Barpeithes pellucidus from one population in Slovakia. The number of B-chromosomes ranged from one to six per cell and they paired with the sex heterochromosomes in the first meiotic metaphase and rarely with the autosomes. In metaphase I one B-chromosome was always associated with X chromosome forming a tripartite complex. The XyBp was easily recognizable as a complex of three chromosomes in a parachute association The size of the B chromosomes was approximately the same or a little smaller than that of the y heterochromosome which was the smallest element of the regular chromosome set. Their staining intensity seems to be similar to that of the autosomes and sex chromosomes, respectively. The behaviour of B-chromosomes during mitosis and meiosis in weevils is briefly discussed. PMID- 16212111 TI - Karyotype study on pseudoscorpions of the genus Lasiochernes Beier (Pseudoscorpiones, Chernetidae). AB - Karyotypes of the genus Lasiochernes (Pseudoscorpiones, Chernetidae) are studied for the first time. The diploid chromosome numbers of males were found to be 2n = 61 in L. pilosus, 2n = 69 in L. siculus and 2n = 73 in L. cretonatus. Karyotypes of all species mainly consist of biarmed chromosomes; the sex chromosome system is XO. Remarkably, the X chromosome displays partial (L. cretonatus), or even total (L. pilosus), negative heteropycnosis during the spermatogonial metaphase. PMID- 16212112 TI - Intra individual variation in the number of B chromosomes in the yellow-necked mouse, Apodemus flavicollis (Mammalia, Rodentia). AB - The yellow-necked mouse, Apodemus flavicollis, is characterized by a frequent occurrence of B chromosomes. The frequency of intra individual mosaicism of Bs was studied in 995 animals collected at six localities in Serbia. It was found that 329 (33.06%) possessed B chromosomes. Among these, 87 animals (26.44%) were mosaics. A total of 32 mosaic animals with more than one B chromosome were analyzed for distribution of Bs which was found to be quite different between groups of animals with different numbers of Bs and increases with their number. The frequency of mosaics differs between localities and ranges from 0.22 to 0.55. PMID- 16212113 TI - The uptake and accumulation of iron by the intestinal bacterium Desulfotomaculum acetoxidans DSM 771. AB - Sulfate-reducing bacteria (e.g. Desulfotomaculum acetoxidans) exist in animal intestine. These bacteria are able to bind heavy metals (e.g. cadmium or lead). Comparative investigations on the composition of cellular walls of Desulfotomaculum acetoxidans--depending on the initial Fe2+ supplement in the medium (7.5, 57.5 and 507.5 M) were performed. Iron(II) was cumulated as FeS or as pyrite (FeS2). However, if the initial amount of iron was higher, its majority (46% 85%) was transported onto the membrane. It was determined that the siderophore found in Desulfotomaculum acetoxidans was deferroxamine as in animals. PMID- 16212114 TI - RAPD-PCR analysis of various goose populations. AB - The aim of this study was to genetically analyse by the RAPD-PCR method four indigenous Polish goose breeds, Kartuska (Ka), Lubelska (Lu), Kielecka (Ki) and Podkarpacka (Pd), in order to determine the band-sharing frequency as well as bands characteristic of the evaluated breeds. The birds were maintained as conservative flocks, accounting for a reserve of genetic resources. A total of 102 scorable bands were obtained, their number ranging from 0 to 8, depending on one of seven primers used and the group of birds analyzed, within a mean of 3.64. For each genetic group specific bands with given primers were obtained, suggesting their potential for use as population-specific markers, especially in ex-situ conservation methods. The results also suggest that keeping endangered geese as separate flocks is relevant for their preservation. PMID- 16212115 TI - Molecular polymorphism of strains within Paramecium septaurelia (Ciliophora, Oligohymenophorea). AB - RAPD-PCR fingerprinting and ARDRA riboprinting revealed polymorphism within P. septaurelia strains from Russia (4 strains from Lower Volga Basin), and one strain from USA, Florida. However, the first method showed the existence of four RAPD genotypes while the second revealed only two groups of strains with different band patterns. All studied strains had a high percentage of surviving hybrid clones in the inter-strain crosses, with little differentiation of strains within species. Intra-species differentiation of strains in RAPD band patterns may be connected with the degree of inbreeding for the studied species. Species of the P. aurelia complex can be arranged according to the degree of inbreeding characteristic for each, which is correlated with the degree of DNA polymorphism revealed by the RAPD method from extreme inbreeders (e.g. P. sexaurelia), moderate inbreeders (e.g. P. triaurelia) to weak inbreeders (e.g. P. pentaurelia). P. septaurelia of the P. aurelia complex should be included in the group of extreme inbreeder. PMID- 16212116 TI - A study of the anatomy, histology and ultrastructure of the digestive tract of Orthrias angorae Steindachner, 1897. AB - The anatomy, histology and ultrastructure of the digestive tract of Orthrias angorae (Steindachner, 1897) were investigated using light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The histological structure consists of four layers: mucosa, submucosa, muscularis and serosa. The esophageal mucosa consists of undifferentiated basal epithelial cells, mucous cells and surface epithelial cells. It was observed that the J shaped stomach had a meshwork of folds in the cardiac region, and longitudinal folds in the fundic and pyloric regions. A single layer of columnar cells, PAS positive only in their apical portions, forms the epithelium. The convoluted tube shape intestine is lined by simple columnar epithelial cells, which have microvilli at the apical surface. The wall of the esophagus and stomach are thicker than that of the intestine because of the thick muscle layer. There were numerous goblet cells in the intestine. There were numerous gastric glands in the submucosa layer ofthe cardiac stomach, but none were present in the pyloric region of the stomach. There were no pyloric caeca between the stomach and intestine. The enterocytes with microvilli contained rough endoplasmic reticulum, ribosomes and rounded bodies, and the gastric cells contained a well-developed Golgi apparatus. PMID- 16212117 TI - Effect of electric power network frequency magnetic field on embryonic development of Ascaris suum (Nematoda). AB - Fertilised Ascaris suum eggs were subjected to an alternating electromagnetic field of frequency 50 Hz and density 2 mT for 60 days. The developing embryos in both control and experimental cultures were examined daily under a microscope. The experiment resulted in an accelerated rate of embryogenesis in the eggs incubated in the electromagnetic field, higher rates of malformed embryos as well as much higher mortality rate of L2 larvae. PMID- 16212118 TI - Resistin: can we resist its role in insulin resistance? PMID- 16212119 TI - Serum resistin, adiposity and insulin resistance in Saudi women with type 2 diabetes mellitus. AB - BACKGROUND: The role of adipocyte hormones in modulating insulin sensitivity and glucose tolerance are of increasing interest and importance in studies of type 2 diabetes mellitus. Recently a unique signaling molecule, resistin, has been proposed as playing a role in the pathogenesis of obesity-related insulin resistance, but its relevance to human diabetes remains uncertain. Therefore, we assessed the relationship between serum resistin concentrations and insulin resistance in lean, overweight and obese (OW/OB) non-diabetic and diabetic Saudi women. SUBJECTS AND METHODS: We measured fasting serum resistin levels in 44 diabetic women with a mean body mass index (BMI) of 31.82 +/- 4.35 kg/m2, 21 OW/OB non-diabetic women with a mean BMI 30.71 +/- 3.42 kg/m2 and in 24 lean women with a mean BMI of 23.33 +/- 1.24 kg/m2. Insulin resistance was assessed using the homeostasis model assessment for insulin resistance formula derived from fasting insulin and glucose levels. RESULTS: The concentrations of fasting serum resistin showed significant differences among the three groups (P<0.001). Mean serum resistin concentrations increased from lean (11.59 +/- 2.08) to OW/OB non-diabetic (16.29 +/- 2.29) to diabetic (19.42 +/- 3.60 ng/mL) women. Significantly higher levels of glucose (P<0.001) and values for the homeostasis model assessment ratio (HOMA-R) (P<0.01) occurred in the diabetic compared to the lean and OW/OB non-diabetic subjects. Furthermore, resistin correlated significantly and positively with hip circumferences (r=0.39, P=0.039), weight (r=0.51, P=0.005), insulin (r=0.40, P=0.033), HOMA-R (r=0.49, P=0.007) and glucose (r=0.39, P=0.038) in diabetic women. In OW/OB non-diabetic subjects, resistin correlated with insulin (r=0.59, P=0.015) and HOMA-R (r=0.616, P=0.011). No correlation was observed with glucose, height, hip, waist, weight, and waist hip ratio (WHR) in the lean and OW/OB non-diabetic groups. CONCLUSION: Resistin concentrations are elevated in patients with type 2 diabetes and are associated with obesity and insulin resistance. These data indicate that resistin might be involved in the development of diabetes in humans. PMID- 16212120 TI - Primary breast lymphoma: a pooled analysis of prognostic factors and survival in 93 cases. AB - BACKGROUND: Primary breast lymphoma is a rare disease. The small number of patients and the paucity of data make large-series studies difficult. We conducted a pooled analysis to evaluate the treatment outcome and prognostic factors in patients with primary breast lymphoma. METHODS: In a search of PUBMED and MEDLINE we found 7 observational studies with 93 patientsthatwere eligible for inclusion. Treatments included single therapy or combined surgery, chemotherapy and radiotherapy. We analyzed the correlation between treatment protocols, tumor relapse and survival. Histopathology and cancer stage were analyzed to evaluate their significance in treatment outcome. RESULTS: All 93 patients were female, with a mean age of 57 years. The histopathology of 63 patients (68%) was diffuse large cell lymphoma. According to Ann Arbor classification, 57% were stage I, 23% were stage II, 4% were stage III, and 16% were stage IV. Thirteen percent received surgery alone, 27% received chemotherapy alone, 7% received radiotherapy alone, 10% received surgery and chemotherapy, 10% received surgery and radiotherapy, 22% received chemotherapy and radiotherapy, and 11% received surgery combined with chemotherapy and radiotherapy. With a median follow-up duration of 34 months (mean, 53 months), 48% had relapse of disease, 50% had no relapse, while 2% had disease progression. The mean time to first tumor relapse after treatment was 20 months. The 3-year and 5-year overall survival rates were 70% and 56%, respectively. Radiotherapy was a significant prognostic factor predicting tumor relapse (P=0.044). Tumor stage was a significant prognostic factor affecting overall survival, disease-free survival and disease-specific survival (P=0.0231, 0.0015, 0.0124, respectively). CONCLUSION: With a 3-year overall survival rate of 70%, the high relapse rate of 48% is a cause for concern. Patients who received chemotherapy and radiotherapy had better survival outcome and a lower relapse rate. We suggestthat chemotherapy and radiotherapy be the initial treatment for patients with primary breast lymphoma. PMID- 16212121 TI - Knowledge of healthy diets among adolescents in eastern Saudi Arabia. AB - BACKGROUND: Bad dietary habits, such as eating high-fat/high-energy food, can contribute to obesity in adolescents, which tends to persist into adulthood. The objective of this study was to determine the level and sources of knowledge about foods and healthy diets among male and female adolescents. METHODS: This was a cross-sectional study of a sample of male (n=1240) and female (n=1331) adolescents from third grade, intermediate and all three grades of secondary school students in the Al-Khobar Area, eastern province of Saudi Arabia. A self administered questionnaire was used to collect information. RESULTS: Approximately 51% of the male and 65% of the female students recognized unsaturated fats as healthy foods. However, 10% of the males and 8% of females reported saturated fats as healthy food items. About 49% of the males and 66% of the females correctly defined cholesterol. Dietary knowledge of both male and female students on the dangers of unhealthy foods and the benefits of fiber-rich diets was found to be unsatisfactory. The main sources of knowledge about health and disease reported by the male and female respondents were television (58% and 61%, respectively), magazines (31% and 39%) and daily newspaper (33% and 34%). Primary health care centers (PHCCs) staff were the least source of knowledge (17% and 16%). CONCLUSIONS: Knowledge of healthy diets among school students was inadequate. It is recommended that health education and information about healthy eating habits and lifestyle be included in school curricula. PMID- 16212123 TI - Prevalence of methicillin-resistant and methicillin-sensitive Staphylococcus aureus nasal colonization among patients at the time of admission to the hospital. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is an important agent of hospital-acquired infection. The mode of entry of MRSA in the hospital might be on admission of patients with MRSA infection or nasal colonization. The present study was undertaken to determine the prevalence of MRSA nasal colonization among patients on admission to hospital. METHODS: Six hundred patients were screened for nasal colonization of MRSA on admission to hospital. Nasal swabs were cultured on salt mannitol agar and blood agar. Age, sex, previous admission to hospital and antibiotic therapy were recorded. RESULTS: S. aureus was isolated from the nasal swabs of 122 patients (20.2%) on admission to hospital. MRSA was isolated from 7 patients (1.1%) and methicillin-sensitive S. aureus (MSSA) from 115 patients (19.1%). Nasal colonization of S. aureus was higher in younger and elderly patients and significantly higher colonization was observed among females. The MRSA strains isolated from nasal swabs had a different antibiotic susceptibility pattern than those isolated from patients having hospital-acquired MRSA infection. Previous admissions to hospital, underlying disease antibiotic therapy were not risk factors for MRSA nasal colonization. CONCLUSION: MRSA nasal colonization of patients on admission to hospital is low in this region. The screening of every new admission would not be cost effective, but patients transferred form other institutions should be screened for MRSA. Standard infection control precautions should be strictly implemented to prevent the spread and control of MRSA infections. PMID- 16212122 TI - Seroprevalence of Helicobacter pylori among children in Sana'a, Yemen. AB - BACKGROUND: Helicobacter pylori infection is primarily acquired in early childhood. Its transmission routes are debated. The aims of this study were to determine the seroprevalence of anti-H. pylori immunoglobulin G (IgG) in Yemeni children under 10 years of age, the potential risk factors for contracting H. pylori infection and co-infection of H. pylori with intestinal parasites. METHODS: Enzyme-labeled immunosorbent assay was used to determine the H. pylori prevalence rate among 572 healthy volunteers aged less than 10 years. Formalin ether concentration methods were used to test the prevalence of intestinal parasites (intestinal roundworms and tapeworms). In addition, we interviewed participants regarding potential risk factors for contracting H. pylori infection. RESULTS: The seroprevalence of H. pylori antibodies was 9%. The prevalence according to age varied from 0% in children under 2 years to 12.5% in age group 9-10 years. There was a correlation between the amounts of positive antibodies and increasing age. The prevalence rate of H. pylori antibodies was also significantly associated with the practice of drinking water from reused plastic jerry cans, with poor mouth hygiene and with co-infection by intestinal parasites. CONCLUSION: The prevalence of H. pylori antibodies in Yemen among children under 10 years of age is higher than that reported from other regions for the same age groups. Yemen shares some but not all potential risk factors for H. pylori infection with countries in which similar socioeconomic conditions are found. A possible way of eliminating H. pylori from the population would be via public health measures, i.e. preventing the reuse of plastic jerry cans, and improving sanitation and the standard of living. PMID- 16212124 TI - Rodent control operations against zoonotic cutaneous leishmaniasis in rural Iran. AB - BACKGROUND: Zoonotic cutaneous leishmaniasis (ZCL) is a serious and increasing public health problem in many rural areas of Iran. Previous studies showed that rodent control operations are effective in destroying rodents and reducing the incidence of ZCL on a small scale and in special circumstances. The objective of this study was to determine suitable timing for rodent control operations in reducing the incidence of ZCL in an endemic area in Badrood, Iran. METHODS: We conducted rodent control operations by baiting rodent holes using zinc phosphide once a month in May, June, July and September in 1997 within a 500-meter circle of houses in the intervention area. From 1999 to 2002, the numbers of active rodent holes were counted in May and October in the intervention and control areas. When the numbers of rodent holes increased to 30% or more of the number before the first baiting in May 1997, holes in the intervention area were baited again with zinc phosphide in the intervention area; no baiting was done in the control village. Case findings were done by house-to-house visits once every season during 2000 to 2002. RESULTS: Changes in the number of rodent holes over time in the intervention and control villages were statistically significant (P<0.000001). There were also significant differences in the incidence of ZCL between the intervention and control villages (P<0.005) during 2000 to 2002. The incidence of ZCLwas unchanged in the years 2000 and 2001, but increased in one village in 2002 after an increase in the number of rodent holes. CONCLUSION: We suggest that rodent control operations using zinc phosphide be done within a 500 meter circle of houses once every two years before the beginning of the active season of sandflies, which begins in late April. PMID- 16212125 TI - Drug-resistant pulmonary tuberculosis in western Turkey: prevalence, clinical characteristics and treatment outcome. AB - BACKGROUND: Although high antituberculosis (anti-TB) drug resistance rates have been reported in Turkey, the clinical characteristics and implications for the outcome of anti-TB treatment have not been fully investigated. We determined the prevalence of anti-TB drug resistance and examined demographic data, clinical characteristics and treatment outcome in relation to patterns of resistance. METHODS: From the TB case registry of a university hospital and the two largest dispensaries in Manisa city, we identified all pulmonary TB cases with a culture proven definitive diagnosis and antimicrobial susceptibility results for a 7-year period. We collected and analyzed demographic and clinical data and information on treatment outcome for those cases in relationship to anti-TB drug resistance. RESULTS: Of 355 M. tuberculosis strains, 71.5% were susceptible to streptomycin, isoniazid, rifampicin and ethambutol. Any drug resistance and multi-drug resistance (MDR) rates were 21.1% and 7.3% and were higher in males (53% and 9%, respectively) than in females (22% and 1%, respectively). Drug resistance was significantly higher in old cases (acquired drug resistance) vs new cases (primary drug resistance), and was associated with treatmentfailure (P<0.001). The prevalence of MDR was significantly higher in the old cases (22.4%) than in the new cases (4.4%) (P<0.001). Symptoms, radiographic findings, associated diseases, and sputum smear positivity were unrelated to the development of resistance. The prevalence of any drug resistance and MDR was significantly higher in those with treatment failure than in patients with treatment success. CONCLUSION: High resistance rates, particularly for acquired MDR, indicate a need for improvement in the TB control programme in our region. PMID- 16212126 TI - Histiocytic necrotising lymphadenitis (Kikuchi-Fujimoto disease) in Saudi Arabia: clinicopathology and immunohistochemistry. AB - BACKGROUND: Kikuchi-Fujimoto disease (KFD) is a rare entity of uncertain cause that commonly presents as a benign self-limiting disease of unknown origin. The objective of this study was to document the clinical features, mode of presentation, histopathological and immunohisto-chemical (IHC) features of KFD at our institutions since little is known about this disease in our region. METHODS: We reviewed the histopathological reports of all lymph nodes resected at or referred to King Abdulaziz University Hospital between 1990 and 2003 and King Faisal Specialist Hospital & Research Center, Jeddah, Kingdom of Saudi Arabia between 2000 and 2003. All cases diagnosed as KFD were identified and the histological slides and clinical data were reviewed. IHC was performed for the proliferative marker Ki-67 and the apoptosis-related markers Bcl-2 and p53. RESULTS: In 2500 lymph node biopsies, 15 cases were diagnosed as KFD. The female to male ratio was 2.7:1. One patient presented with axillary lymphadenopathy and the others presented with cervical lymphadenopathy. Ages averaged 29 years and ranged from 13 to 46 years. There was no recurrence of the lymphadenopathy over 1 to 10 years of follow up. Bcl-2 and p53 were negative and Ki-67 was positive in 11 of 15 cases. CONCLUSION: The results support earlier findings that KFD is a self-limiting disorder that requires no specific management. We suggest a clinical follow-up for several years. The female predominance was striking. Apoptosis-regulating proteins are not helpful in the diagnosis. KFD usually expressed the proliferation-associated nuclear antigen Ki-67. Increased awareness of KFD will minimize the risk of confusing this entity with malignant lymphoma or other serious conditions. PMID- 16212127 TI - Measles in Saudi Arabia: from control to elimination. AB - This article describesthe tremendous efforts made in the field of measles immunization in Saudi Arabia in the past 20 years, from the control phase to the elimination phase. Mandatory measles vaccination with one-dose Schwartz vaccine was introduced in 1982 by a royal decree, a step aimed at increasing vaccine coverage. In 1991, a two-dose schedule was implemented using Edmonston-Zagreb measles vaccine, with a first dose at 6 months to protect children younger than 9 months and a second dose of MMR at 12 months of age to protect those who did not respond to the first dose. A marked reduction in the epidemic peak and a shift of infection to older age were noticed. But the same data showed that 50% of measles cases in the 1- to 4-year age group occurred in vaccinated children. In 1998, with the start of elimination phase, an MMR campaign was launched in two phases, targeting school children in 1998 (secondary schools) and in 2000 (primary and intermediate schools). Evaluation of the MMR campaign and surveillance data was reflected in the measles immunization policy by shifting the age of measles immunization to 12 months and to preschool using the two-dose MMR schedule. PMID- 16212129 TI - Pulmonary edema complicating ovarian hyperstimulation syndrome: low-pressure edema, high-pressure edema, or mixed edema? PMID- 16212130 TI - Unusual management of a non-communicating uterine horn. PMID- 16212128 TI - Unresolved issues and current concepts in management of primary glomerulonephritis. AB - The successful treatment of primary glomerulonephritis (GN) presenting with nephrotic syndrome in adults depends heavily on an accurate diagnosis. A successful diagnosis depends on a correct approach, combining light microscopy, immunofluorescence, and other special staining of renal biopsy material examined by a trained nephropathologist. A good clinical history and serological tests easily rule out possible secondary causes (for example, infection, autoimmune, metabolic or toxic) in most cases. Unfortunately, these procedures are not put into practice in most cases in developing countries, resulting in missed diagnosis and unnecessary steroid and immunosuppressant therapy with its inherent morbidity. Following the emergence of IgA and IgM nephropathies as very common forms of glomerular disorders in some countries, immunofluorescence has become absolutely necessary for their diagnosis. Moreover, a recent meta-analysis has defined different treatment protocols for minimal change nephropathy, focal segmental glomerulosclerosis, membranous nephropathy, and IgA nephropathy for a better outcome. This article emphasizes and elaborates on these issues for proper management of primary GN. PMID- 16212131 TI - Post-traumatic cutaneous mucormycosis in an immunocompetent patient. PMID- 16212132 TI - Blind-ending ureteral duplication with calculi. PMID- 16212133 TI - A three-year old girl with Down's syndrome and an abnormal finding in the chest. PMID- 16212134 TI - Re: EIF2AK3 mutations in patients with Wolcott-Rallison syndrome. PMID- 16212135 TI - Isolated double gastric rupture as a result of blunt abdominal trauma. PMID- 16212136 TI - Sir Charles Sherrington--"The William Harvey of the nervous system". PMID- 16212137 TI - Cardiovascular risk linked to chronic kidney disease--but who actually has chronic kidney disease? PMID- 16212138 TI - Cardiovascular risk factor profiles and kidney function stage in the US general population: the NHANES III study. AB - OBJECTIVE: To determine the prevalence of cardiovascular risk factors in the general population based on kidney function. SUBJECTS AND METHODS: We retrospectively analyzed data from the Third National Health and Nutrition Examination Survey of noninstitutionalized US adults, which was conducted from 1988 to 1994. Data were gathered on 9 cardiovascular risk factors (smoking; obesity; hypertension; high total cholesterol, C-reactive protein, glycosylated hemoglobin, and homocysteine levels; low hemoglobin level; and high urinary albumin-to-creatinine ratio) and estimated glomerular filtration rate (GFR). RESULTS: For the 15,837 subjects, the estimated GFR was at least 90 mL/min per 1.73 m2 (normal) in 65.4%, 60 to 89.9 mL/ min per 1.73 m2 (stage 2 kidney function) in 27.9%, 30 to 59.9 mL/min per 1.73 m2 (stage 3 kidney function) in 6.2%, and less than 30 mL/min per 1.73 m2 (stages 4 and 5 kidney function) In 0.5%. The number of cardiovascular risk factors Increased with stage of kidney dysfunction. Of subjects with a normal GFR, 30.4% had no risk factors, 34.9% had 1 risk factor, and 34.7% had 2 or more risk factors. Of subjects with stage 2 kidney function, 24.8% had no risk factors, 30.3% had 1 risk factor, and 44.9% had 2 or more risk factors. Of subjects with stage 3 kidney function, 1.4% had no risk factors, 14.9% had 1 risk factor, and 83.6% had 2 or more risk factors. All subjects with stages 4 and 5 kidney function had 2 or more risk factors. After covariate adjustment, odds ratios for having an estimated GFR lower than 60 mL/min per 1.73 m2 were 1, 3.7 (95% confidence interval, 1.2-11.3), and 10.4 (95% confidence interval, 3.9-27.8) times greater in subjects with 0, 1, and 2 or more cardiovascular risk factors, respectively (P<.001). CONCLUSION: Persons with chronic kidney disease are much more likely to need multiple cardiovascular risk factor interventions than those without chronic kidney disease. PMID- 16212139 TI - Upper gastrointestinal tract safety of daily oral risedronate in patients taking NSAIDs: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate the frequency of upper gastrointestinal (GI) tract adverse events associated with risedronate during two (2-year) randomized, double-blind, parallel-group, placebo-controlled studies. PATIENTS AND METHODS: Male and female patients aged 40 to 80 years with mild to moderate medial-compartment knee osteoarthritis were enrolled. Data were pooled and analyzed for risedronate at 5 mg and at 15 mg once daily and compared with placebo. The results of the once weekly dosages (35 or 50 mg) were assessed separately. RESULTS: A total of 2483 patients were randomized: 622 to placebo, 628 to risedronate at 5 mg/d, 609 to risedronate at 15 mg/d, 310 to risedronate at 35 mg once weekly, and 314 to risedronate at 50 mg once weekly. During the study, 77% of patients were regular nonsteroidal anti-inflammatory drug (NSAID) and/or analgesic users (defined as those who took medication 23 days per week), and 68% were regular NSAID users. The number of upper GI tract adverse events was similar between treatment groups, with no dose-related response: 161 for placebo, 176 for rlsedronate at 5 mg/d, and 150 for risedronate at 15 mg/d. The time to the first upper GI tract adverse event was similar between treatment groups. There was no difference in the frequency of upper GI tract adverse events In risedronate-treated patients compared with patients who were regular users of NSAIDs or NSAIDs and/or analgesics. Findings were similar for those in the once-weekly risedronate groups. CONCLUSION: The results of this study show that risedronate regimens at 5 mg/d or 15 mg/d as well as once weekly at 35 mg or 50 mg are not associated with an increased frequency of upper GI tract adverse events, even in patients who have an increased risk for such events. PMID- 16212140 TI - Cellular telephone interference with medical equipment. AB - OBJECTIVE: To assess the potential electromagnetic interference (EMI) effects that new or current-generation cellular telephones have on medical devices. MATERIAL AND METHODS: For this study, performed at the Mayo Clinic in Rochester, Minn, between March 9, 2004, and April 24, 2004, we tested 16 different medical devices with 6 cellular telephones to assess the potential for EMI. Two of the medical devices were tested with both new and old interface modules. The 6 cellular telephones chosen represent the different cellular technology protocols in use: Code Division Multiple Access (2 models), Global System for Mobile communications, Integrated Digital Enhanced Network, Time Division Multiple Access, and analog. The cellular telephones were tested when operating at or near their maximum power output. The medical devices, connected to clinical simulators during testing, were monitored by observing the device displays and alarms. RESULTS: Of 510 tests performed, the incidence of clinically important interference was 1.2%; EMI was Induced in 108 tests (21.2%). Interference occurred in 7 (44%) of the 16 devices tested. CONCLUSIONS: Cellular telephones can interfere with medical equipment. Technology changes in both cellular telephones and medical equipment may continue to mitigate or may worsen clinically relevant interference. Compared with cellular telephones tested in previous studies, those currently in use must be closer to medical devices before any interference is noticed. However, periodic testing of cellular telephones to determine their effects on medical equipment will be required. PMID- 16212141 TI - Sustained efficacy and safety of vardenafil for treatment of erectile dysfunction: a randomized, double-blind, placebo-controlled study. AB - OBJECTIVE: To evaluate the reliability, efficacy, and safety of vardenafil, 10 mg, for patients with erectile dysfunction. PATIENTS AND METHODS: Vardenafil naive patients completed a 4-week treatment-free run-in phase and a 1-week single dose vardenafil (10 mg) open-label challenge phase. Responders to vardenafil in the challenge phase were randomized to 12 weeks of double-blind, fixed-dose treatment with vardenafil at 10 mg or placebo. Diary responses to Sexual Encounter Profile (SEP) questions about erections and attempts at sexual activity were collected after 4, 8, and 12 weeks of randomized treatment. Adverse events were monitored throughout the study. RESULTS: During the open-label challenge phase, the proportions of patients with a first-time success for penetration (SEP2) and maintenance of erection (SEP3) were 87% and 74%, respectively. Of 600 patients challenged with a single dose of vardenafil at 10 mg, 260 were randomized to vardenafil and 263 to placebo. During the double-blind phase, the reliability of penetration and maintenance rates for patients successful during the challenge phase were significantly greater with vardenafil compared with placebo (83.4% vs 55.8% [SEP2] and 76.6% vs 42.1% [SEP3], respectively). At week 12, patients in the vardenafil group had a consistently higher least squares mean (SE) on the erectile function domain score of the International Index of Erectile Function than patients in the placebo group (23.5 [0.4] vs 15.8 [0.4], respectively [last observation carried forward]) and a greater proportion of positive responses to the Global Assessment Question (80.8% vs 32.3%, respectively [last observation carried forward]) at each assessment (Pc.001). Vardenafil was generally well tolerated; most adverse events were mild to moderate, with headache and flushing reported most frequently. CONCLUSION: During this 12-week study, vardenafil produced consistently higher reliability of penetration and maintenance of erection rates compared to placebo and was generally well tolerated in patients with erectile dysfunction. PMID- 16212142 TI - Chronic nitrofurantoin-induced lung disease. AB - OBJECTIVE: To reassess the clinical and radiological features of chronic nitrofurantoin-induced lung disease and eventual clinical outcome. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 18 patients with chronic nitrofurantoin-induced lung disease who were seen at the Mayo Clinic in Rochester, Minn, from January 1, 1997, to December 31, 2002. RESULTS: The median age of the 18 patients was 72 years (range, 47-90 years) at the time of diagnosis; 17 (94%) were women. Onset of symptoms occurred after a median interval of 23 months (range, 10-144 months) following the initiation of nitrofurantoin therapy for the prevention of recurrent urinary tract infections. All patients presented with persistent dyspnea and cough associated with lung infiltrates detected on chest radiography. Ten computed tomograms were available for review and revealed bilateral areas of ground-glass opacities in all cases and showed subpleural Irregular linear opacities and patchy consolidation in some cases. Nitrofurantoin therapy was discontinued in all patients, and most improved subsequently; 9 patients received corticosteroid therapy. CONCLUSIONS: Chronic nitrofurantoin-induced lung disease is seen predominantly in older women who present with respiratory symptoms after a year or more of nitrofurantoin therapy. Associated radiological features are relatively nonspecific but usually include bilateral areas of ground-glass opacities on computed tomography of the chest. Cessation of nitrofurantoin therapy leads to improvement and suffices in the management of some patients, although corticosteroid therapy may be helpful in those more severely affected. PMID- 16212143 TI - Spectrum of pneumonia in the current era of liver transplantation and its effect on survival. AB - OBJECTIVE: To examine the frequency and microbial pattern of pneumonia and its effect on survival in the current era of orthotopic liver transplantation (OLT). PATIENTS AND METHODS: At the Mayo Clinic in Jacksonville, Fla, the medical records of consecutive patients who underwent their first OLT between February 1998 and January 2001 were retrospectively reviewed through the end of the first year posttransplantation. RESULTS: Of 401 study patients, 20 developed pneumonia; estimates of incidence with corresponding 95% confidence interval (CI) at 1 and 12 months were 3% (1%-5%) and 5% (3%-7%), respectively. Pseudomonas aeruginosa was the predominant microorganism identified (in 8 of 14 patients) during the first month after transplantation. Between the second and sixth months, 2 of the 4 cases of pneumonia were due to fungal infections of Aspergillus fumigatus. Cytomegalovirus was associated with Aspergillus in 1 patient. No other viral or Pneumocystis carnil pneumonia was diagnosed. There were only 2 cases of pneumonia between 7 months and 1 year after transplantation, neither of which was fungal. Approximately 40% (95% CI, 14%-58%) of patients with pneumonia died within 1 month after diagnosis. The relative risk of mortality in the first month after onset of pneumonia was estimated to be 24 (95% CI, 10-54), which is strong evidence of increased risk of mortality with pneumonia (P<0.001). CONCLUSIONS: Pneumonia appears to occur less often after OLT than previously reported but still has a substantial negative effect on survival. In the early period after OLT, P. aeruginosa continues to be the predominant organism causing pneumonia. PMID- 16212144 TI - Cardiovascular toxicities of performance-enhancing substances in sports. AB - Athletes commonly use drugs and dietary supplements to improve athletic performance or to assist with weight loss. Some of these substances are obtainable by prescription or by illegal means; others are marketed as supplements, vitamins, or minerals. Nutritional supplements are protected from Food and Drug Administration regulation by the 1994 US Dietary Supplement Health and Education Act, and manufacturers are not required to demonstrate proof of efficacy or safety. Furthermore, the Food and Drug Administration lacks a regulatory body to evaluate such products for purity. Existing scientific data, which consist of case reports and clinical observations, describe serious cardiovascular adverse effects from use of performance-enhancing substances, including sudden death. Although mounting evidence led to the recent ban of ephedra (ma huang), other performance-enhancing substances continue to be used frequently at all levels, from elementary school children to professional athletes. Thus, although the potential for cardiovascular injury is great, few appropriately designed studies have been conducted to assess the benefits and risks of using performance-enhancing substances. We performed an exhaustive OVID MEDLINE search to Identify all existing scientific data, review articles, case reports, and clinical observations that address this subject. In this review, we examine the current evidence regarding cardiovascular risk for persons using anabolic-androgenic steroids including 2 synthetic substances, tetrahydrogestrinone and androstenedione (andro), stimulants such as ephedra, and nonsteroidal agents such as recombinant human erythropoietin, human growth hormone, creatine, and beta-hydroxy-beta-methylbutyrate. PMID- 16212145 TI - Psychological effect, pathophysiology, and management of androgenetic alopecia in men. AB - Androgenetic alopecia In men, or male pattern baldness, is recognized increasingly as a physically and psychologically harmful medical condition that can be managed effectively by generalist clinicians. This article discusses the clinical manifestations, epidemiology, physical and psychosocial importance, pathophysiology, diagnosis, and management of androgenetic alopecia in men. Androgenetic alopecia affects at least half of white men by the age of 50 years. Although androgenetic alopecia does not appear to cause direct physical harm, hair loss can result in physical harm because hair protects against sunburn, cold, mechanical injury, and ultraviolet light. Hair loss also can psychologically affect the balding individual and can Influence others' perceptions of him. A progressive condition, male pattern baldness is known to depend on the presence of the androgen dihydrotestosterone and on a genetic predisposition for this condition, but its pathophysiology has not been elucidated fully. Pharmacotherapy, hair transplantation, and cosmetic aids have been used to manage male pattern baldness. Two US Food and Drug Administration approved hair-loss pharmacotherapies-the potassium channel opener minoxidil and the dihydrotestosterone synthesis inhibitor finasteride--are safe and effective for controlling male pattern baldness with long-term daily use. Regardless of which treatment modality is chosen for male pattern baldness, defining and addressing the patient's expectations regarding therapy are paramount in determining outcome. PMID- 16212146 TI - Neuron-astrocyte interactions: partnership for normal function and disease in the central nervous system. AB - Interactions between neurons and astrocytes are critical for signaling, energy metabolism, extracellular ion homeostasis, volume regulation, and neuroprotection in the central nervous system. Astrocytes face the synapses, send end-foot processes that enwrap the brain capillaries, and form an extensive network interconnected by gap junctions. Astrocytes express several membrane proteins and enzymes that are critical for uptake of glutamate at the synapses, ammonia detoxification, buffering of extracellular K+, and volume regulation. They also participate in detection, propagation, and modulation of excitatory synaptic signals, provide metabolic support to the active neurons, and contribute to functional hyperemia in the active brain tissue. Disturbances of these neuron astrocyte interactions are likely to play an important role in neurologic disorders including cerebral ischemia, neurodegeneration, migraine, cerebral edema, and hepatic encephalopathy. PMID- 16212147 TI - Principles and process in the development of the Mayo Clinic's individual and institutional conflict of interest policy. AB - In 1995, federal regulations required all academic medical centers to implement policies to manage individual financial conflict of interest. At the Mayo Clinic, all staff are salaried, and all medically related intellectual property from the staff belongs to the clinic. Hence, it was necessary to develop a policy for institutional conflict of interest to complement the policy for individual conflicts of interest. This article addresses the principles and process that led to the development of the Mayo Clinic's policies that guide the management of conflict of interest of individuals and of the institution. Empowered by the Bayh Dole Act, the Mayo Clinic participates in technology transfer through its entity Mayo Medical Ventures. Individual conflicts of interest arising from such technology transfer are associated with Institutional conflicts because all individual intellectual property belongs to the institution, per clinic policy. This policy addresses conflicts of interest that arise in research, leadership, clinical practice, investments, and purchasing. Associated with the statutory annual disclosure on personal consulting and other relationships with Industry, which are guided by federal regulations, all research protocols or grant applications require financial disclosure on initial submission and in annual progress reports. The clinic's Conflict of Interest Review Board was established to review each disclosure and recommend management of individual and institutional conflicts of interest according to policy. PMID- 16212148 TI - The promise and risk of academic medicine. PMID- 16212149 TI - Evaluation and medical management of benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is common among aging men. Untreated BPH may lead to complications including urinary tract infection, acute urinary retention, and obstructive nephropathy. Diagnosing BPH can be challenging because lower urinary tract symptoms are found in conditions other than BPH, and prostate size correlates poorly with symptoms of obstruction. Nonetheless, a careful medical history and physical examination, along with prudent use of diagnostic tests, can yield an accurate diagnosis. We review the evaluation of men with suspected BPH and indications for referral to a urologist for invasive therapy. We also review supporting evidence and treatment considerations for saw palmetto and the 2 major classes of prescription medications, alpha1-adrenergic antagonists and 5alpha reductase inhibitors. PMID- 16212150 TI - 61-year-old woman with knee pain and confusion. PMID- 16212151 TI - Clopidogrel hypersensitivity syndrome with rash, fever, and neutropenia. AB - We report a case of a severe hypersensitivity syndrome reaction to clopidogrel that resolved with dechallenge and recurred on rechallenge. The reaction included neutropenia, rash, fever, tachycardia, nausea, and vomiting. This presentation is very similar to the frequently reported hypersensitivity syndrome reactions to the structurally similar drug ticlopidine and is very similar to a case involving clopidogrel reported previously in this Journal. We could find no other reports of similar cases related to clopidogrel. PMID- 16212153 TI - Endovascular aortic aneurysm repair in a patient with coronary artery disease. PMID- 16212152 TI - Multiple myeloma: diagnosis and treatment. AB - Major advances have occurred in our understanding of the biology of multiple myeloma (MM) and in its treatment in the past decade. New diagnostic criteria have been developed, and an international Staging System has replaced the Durle Salmon Staging System. It is now possible to classify MM as standard risk or high risk on the basis of specific Independent prognostic factors. The role of single and double autologous stem cell transplantation has been clarified by randomized trials. Most importantly, thalidomide, bortezomib, and lenalidomide have emerged as new active agents and are being incorporated rapidly into the treatment of both newly diagnosed and relapsed MM. The current approach to the diagnosis, prognosis, and management of MM is reviewed. PMID- 16212154 TI - How to interpret and pursue an abnormal complete blood cell count in adults. PMID- 16212156 TI - Paroxysmal nocturnal hemoglobinuria after chloramphenicol therapy. PMID- 16212155 TI - How to interpret and pursue an abnormal complete blood cell count in adults. PMID- 16212157 TI - Accuracy of the Mayo Clinic Patient/Family History Questionnaire. PMID- 16212158 TI - [Mass size distribution of Beijing particulate matters and its inorganic water soluble ions in winter and summer]. AB - To understand one of the important characteristics of Beijing particulate matter pollution, the mass size distribution of particulate matters and its water soluble ions, the particle samples were collected by a MOUDI cascade impactor in Beijing during July 2001, March 2002, July 2002 and January 2003 and every sampling period lasted a week. By analysis, the results indicate that there is "two modes" in the diameter range of the fine particle, one called "condensation mode" and another called "droplet mode", and the reasons of the peak of the accumulation mode appearing in the diameter range of 1 - 1. 8 microm was discussed in this paper. And there was a relatively strong correlation between PM1.8 and PM10 (R2 > 80%), the same to PM(1.8-10) (PM10 minus PM1.8); the ratio of PM1.8s/PM10 varietied between 40% and 60% during the sampling periods; as a result, the mass concentration of PM10 can be decreased by decreasing the concentration of coarse particles; sulfate, nitrate and ammonium are the main component of the inorganic water-soluble ions and the percentage of these three ions in PM1.8 was above 70% when the concentration of fine particle was more than 70 microg x m(-3) and they are the main reason increasing particle concentration. PMID- 16212159 TI - [Dynamics of CO2, CH4 and N2O emission fluxes from mires during freezing and thawing season]. AB - In the Sanjiang Plain, the freezing and thawing phase have 7 - 8 months and play important role in the greenhouse gases emission. The characters of the greenhouse gases emission during freezing and thawing season in the Sanjiang Plain were studied, using the static chamber and gas chromatogram method. The results observed show that there were obvious CH4 and CO2 emissions in winter and the CH4 emission made a relatively large contribution to the total CH4 flux from the different type mires during the winter in the Sanjiang Plain. And there were significantly CH4 and CO2 emission peak values during thawing time. The CH4 and CO2 emissions fluxes from seasonal flooded mire were larger than that from continuously flooded mire during thawing time. On the contrary, the CH4 and CO2 emissions from continuously flooded mire were larger than that from seasonal flooded mire in winter. During thawing, there was exponential relationship between CO2 fluxes and the soil temperature (5cm) (R2 = 0.912, p< 0.001). Meanwhile, CO2 fluxes was obviously correlated with the CH4 emission fluxes (R2 = 0.751, p < 0.001). The mires are N2O sink in winter and gradually become N2O source, with increasing temperature during thawing time. The characters of the greenhouse gases emission, during freezing and thawing in the Sanjiang Plain respond to the microbial activity in winter and the effects of thawing on soil carbon mineralization, nitrification and denitrification. PMID- 16212160 TI - [Contribution of anaerobic oxidation of methane to whole methane oxidation]. AB - The contribution of anaerobic methane oxidation to the whole methane oxidation is investigated by monitoring the activities of anaerobic and aerobic oxidation of methane in paddy soils composed mechanically of <0.02 mm and > 0.02 mm soil granules, contained different water contents, and long-term fertilized organic manure or inorganic nitrogen, phosphorus and potassium fertilizers. The results obtained from various treated soils are relatively identical, which shows that most of the activities of anaerobic methane oxidation is obviously lower than those of aerobic methane oxidation. The contribution of anaerobic methane oxidation was generally lower than 10% of total methane oxidation in paddy soils with different treatments, however, higher than 30% in the soils emerged in water as the soils became anaerobic and methane diffused difficultly into soil, leading to lower activity of aerobic methane oxidation. The long-term fertilization has a significant effect on the activity of aerobic methane oxidation in paddy soil, with the / t / > t0.05, P < 0.05, but a slightly one with ( / t / < t 0.05, p > 0.05), leading to the obvious influence on the emission flux of methane. The contribution rates of anaerobic methane oxidation were lower from 1.31% to 4.14% in the whole methane oxidation. PMID- 16212161 TI - [Measurement method research on trace volatile organic compounds in atmosphere]. AB - An automated accumulation sampling system and a method of two-step Cryo Concentrated System combined with Gas Chromatography/Mass Spectrometry (CCS GC/MS) are introduced. The method is evaluated by a set of special experiments and the results are demonstrated. The lowest limit volume concentrations of the measurement is expanded from 10 (-6) to 10(-12) using CCS-GC/ MS instead of the normal method of GC/MS and the average response factor of 39 object compounds is 2.9 x 10(-12) x A(-1). When injection volume of air sample reaches 1000 cm3, the lowest limit is 7 x 10(-12) - 40 x 10(-12); CCS-GC/MS may identify all the objective chemical species in the atmosphere samples, with average 2.5s bias of retention time for 39 GC peaks. Within 0 - 400 x 10(-9), the concentration of all the objective compounds can be calculated by standard curve individually (average r2 above 0.99) and accurately. The recollect rate is 88% - 111%, at average of 100.8% +/- 5.6%; and the bias of precision is 2% - 14%, at average of 6.6%. PMID- 16212162 TI - [Removal of CO2 from simulated flue gas of power plants by membrane-based gas absorption processes]. AB - Three typical absorbents such as aqueous of aminoacetic acid potassium (AAAP), monoethanolamine (MEA) and methyldiethanolamine(MDEA) are selected to investigate the performance of CO2 separation from flue gas via membrane contactors made of hydrophobic hollow fiber polypropylene porous membrane. Impacts of absorbents, concentrations and flow rates of feeding gas and absorbent solution, cyclic loading of CO2 on the removal rate and the mass transfer velocity of CO2 are discussed. The results demonstrate that the mass transfer velocity was 7.1 mol x (m2 x s)(-1) for 1 mol x L(-1) MEA with flow rate of 0.1 m x s(-1) and flue gas with that of 0.211 m x s(-1). For 1 mol L(-1) AAAP with flow rate of 0.05 m x s( 1) and flue gas of 0.211 m x s(-1), CO2 removal rate (eta) was 93.2 % and eta was 98% for 4 mol x L(-1) AAAP under the same conditions. AAAP being absorbent, eta was higher than 90% in a wider range of concentrations of CO2. It indicates that membrane-based absorption process is a widely-applied and promising way of CO2 removal from flue gas of power plants, which not only appropriates for CO2 removal of flue gas of widely-used PF and NGCC, but also for that of flue gas of IGCC can be utilized widely in future. PMID- 16212163 TI - [Clay ratio as a tracer to assess the origin of soil-derived dust]. AB - The objectives of this research are to characterise the mineralogy of soil derived dust in Northern China and to set up the mineralogical signature to trace their origin. Mineral composition of aerosol particles is investigated at five sites (Aksu, Dunhuang, Yulin, Tongliao and Changwu) during the intensive field campaign period of ACE-Asia. The results show that the Kaolinite (K) to Chlorite (C) ratio is sensitive to the regional origin of Asian dust. Western source areas (represent by Aksu) displayed the lowest K/ C ratio of 0.3 (in average), while it was found that to increase up to 0.70 (in average) as moving towards north source areas (represent by Yulin). By studying transported dust in a depositional area representative of the Chinese Loess Plateau, the usefulness of the K/C ratio to retrieve the origin of the dust by associating it with back air-mass trajectories is checked. Compared the mineralogical data between Asian dust and Sahara dust, it is shown that K/C ratio is also a good signature to identify the source areas on the global scale. PMID- 16212164 TI - [Photocatalytic degradation of low level formaldehyde on TiO2 porous film]. AB - Photocatalytic degradation of low level formaldehyde in the gas phase was studied in a intermittent recirculation photocatalytic reaction system with TiO2 porous film photocatalyst coated on stainless steel mesh. The effects of carrier of the photocatalyst, coating cycle of the photocatalyst, the initial concentration of formaldehyde and the illumination source were investigated respectively. The study of the aging stability of the photocatalyst was carried out as well. Results show that the degradation efficiency of formaldehyde increased with the increase of mesh number. The degradation efficiency also increased with the increase of carrier number, but the influence of carrier number on the degradation efficiexcy was gradually weakened with increasing carrier number. The activity of the photocatalyst firstly increased and then decreased when coating cycle increased from 1 cycle to 6 cycles. When initial concentration was in the range of 1.34 - 10.72 mg/m3, the change of formaldehyde concentration markedly increased with the increase of initial concentration, but the degradation efficiency increased and subsequently decreased with increasing initial concentration. The degradation efficiency with germicidal light was approximately 19.0% higher than that with black light holding same power, and the enhancement in light intensity could approximately increase the degradation efficiency by 20.5%. Finally, the experiment on the longevity of the photocatalyst indicated that the photocatalyst had the good photocatalytic activity after being used for 4 cycles. PMID- 16212165 TI - [Photodegradation of bisphenol A in presence of Suwannee River fulvic acid]. AB - Under simulated solar irradiation, the degradation of bisphenol A (BPA) was studied in the presence of Suwannee River Fulvic Acid (SRFA). The results demonstrate that the photodegradation of BPA followed a pseudo-first-order kinetics and the photodegradation rate increased rapidly with increasing initial concentration of SRFA. Hydroxyl radical, singlet oxygen were found in the SRFA solutions of BPA with molecular probes and the technique of electronic spin resonance. The electronic energy transfer of triplet state fulvic acid was also studied with various aerated conditions. The results showed that the photodegradation of BPA was related with triplet state fulvic acid. The photodegradation products of BPA in the presence of SRFA were identified with GC/MS methods. The photodegradation pathways of BPA were also discussed. PMID- 16212166 TI - [Photoreduction of Se (VI) by marine algae-transitional metals-light system]. AB - Seven marine phytoplankton, including five green algae (Tetraselmis levis, Chlorella autotrophica, Dunaliella salina, Nannochloropsis sp. and Tetraselmis subcordiformis), one diatom (Phaeodactylum tricornutum), one red alga (Porphyridium purpureum), and three usual transitional metals (Fe(III), Cu(II), Mn(II)) were used to make up marine phytoplankton-light or transitional metals light or marine phytoplankton-transitional metals-light system. In such system, Se(VI) could be transformed into Se(IV) by photoreduction. The species transformation of selenium could be photo-induced by redox reaction of transitional metals. The photochemical activity of marine phytoplankton was confirmed for the first time, because marine phytoplankton could adsorb and concentrated of selenium, transitional metals and organic substances (including the exudation of algae, as reducing agent) which redox potentials were changed. The ratios of Se(VI) to Se(IV) were dominated by the species, the concentration of marine phytoplankton and transitional metals, and it could be enhanced through increasing the concentration of marine algae or the combined effect from marine algae and transitional metals. After photoreduction by ternary system, the ratio of Se(VI) to Se(IV) ranges from 1.17 to 2.85, which is close to the actual value in euphotic layer of seawater. The photochemical process that is induced by marine algae and transitional metals dominative the leading effects on the distribution of oxidation states of selenium. PMID- 16212167 TI - [Electrochemical reduction characteristics and mechanism of chlorinated hydrocarbon at the copper electrode]. AB - The electrochemical reduction characteristics of chlorinated hydrocarbons were investigated by applying cyclic voltammetry technique. The reduction mechanism and reactivity of the chlorinated hydrocarbons at the copper electrodes were explored. The relation between the reductive reactivity at the copper electrode and the structures of this kind of compounds was discussed. The experimental results show that chlorinated paraffin hydrocarbons and a portion of chlorinated aromatic hydrocarbons could be reduced directly at the copper electrode; however, chlorinated aromatic hydrocarbons aren't easy to reduced directly at the copper electrode. The results provide a theoretical basis for the catalyzed iron inner electrolysis method. PMID- 16212168 TI - [Reductive dechlorination of trichloroacetic acid by bioelectrochemically catalytic method]. AB - Direct electrochemical behaviors of hemoglobin (Hb) immobilized on carbon nanotube (CNT) modified carbon paste electrode with adsorption were investigated. Cyclic voltammetry of Hb-CNT-modified electrode showed a pair of well-defined and nearly reversible peaks for HbhemeFe(III) /Fe(II) redox couple in pH = 7 PBS buffers. The electrocatalytic behaviors of Hb-CNT-modified electrode for the reductive dechlorination of trichloroacetic acid (TCA) were studied by cyclic voltammmetry and fixed-potential electrolysis technique, and the reductive mechanism of TCA was discussed by analysis of reduction products. The results showed that Hb-CNT-modified electrode possessed good electro-catalytic activity for reduction of TCA and the dechlorination of TCA was stepwise, following the pathway of trichloroacetic--> dichloroacetic--> monochloroacetic--> acetic. The dechlorination of TCA in waster water was investigated using a two-compartment flow reactor with working electrode compartment packed with Hb-CNT-modified graphite electrode. The conversion of TCA was 40.13% with electrolysis for 180 min at - 0.60V (vs. SCE). PMID- 16212169 TI - [Study on dechlorination of p-chlorophenol by Ni/Fe bimetallic particles]. AB - Dechlorination property of p-chlorophenol (p-CP) by Ni/Fe bimetallic particles was studied in this work. The experimental results show that adsorbed hydrogen atom is the main reductive agent for dechlorination of p-CP and chemical reaction at the surface of catalyst is the rate-determing step in this process. The catalyst with mass ratio of 2.96% Ni had the largest specific surface area and also had the highest dechlorination efficiencies (64% in 90 min) under the same conditions. The apparent kinetics of p-CP dechlorination by Ni/Fe was the first order reaction and apparent rate constant (kappa) was proportional to the specific surface of catalyst. The apparent rate constant per specific surface area (kappa') was also calculated (7.61 x 10(-4) min(-1) m(-2)). Temperature was also an important parameter in this system. The dechlorination efficiency was proportional to temperature when it is below 43 degrees C, when temperature exceeded 43 degrees C, increasing temperature had inverse effect on dechlorination efficiency. PMID- 16212170 TI - [Modelling pollutant loads and management alternatives in Jiulong River watershed with AnnAGNPS]. AB - The modelling package Annualized Agricultural Nonpoint Source Model (AnnAGNPS) was used to predict pollutant loads, and simulate catchment processes and management practices in Jiulong River watershed, a medium-sized mountainous watershed in southeast of China. Four typical sub-watersheds were primarily chosen to calibrate AnnAGNPS model by data collected from storm events during the period of April to September, 2003. The model was further validated in the two biggest branches of Jiulong River watershed, i.e. West river and North river by the data regarding climate, and land using condition in 2002 - 2003. The simulation results show that annual total nitrogen load was 24.76kg/(hm2 x a) and 10.28kg/(hm2 x a) in the West river and North river, respectively, and annual total phosphorus load was 0.67 kg/(hm2 x a) and 0.40 kg/(hm2 x a) in the West river and North river, respectively. With the support of AnnAGNPS model, several management alternatives were separately simulated in the typical sub-watersheds, West river and North river. In the specific cell with cell-ID of 92 in Tianbao and Xiandu sub-watershed, after reforesting in sloping field, runoff surface, sediment yield, total nitrogen load and total phosphorus load cut down with 21.6%, 25.9%, 96% and 79.2%, respectively. In West river, with the cultivation plant changing from banana into rice, the total nitrogen, dissolved nitrogen, total phosphorus and dissolved phosphorus cut down with 23.83%, 25.44%, 9.08% and 19.84%, respectively. In North river, when removing all the hoggerys, nitrogen and dissolved nitrogen cut down with 63.54% and 76.92% , respectively. PMID- 16212171 TI - [Phosphorus sorption capacities of the pond sediments in a headstream agricultural watershed]. AB - The land uses in watersheds have important influence on the sediments of lakes and wetlands. Field and laboratory experiments were conducted to research the phosphorus adsorption by the pond sediments in the pond system in a headstream watershed. Being scattered in different ambient land uses and receiving soil particles from various sources, the pond system provides an excellent research example for the phosphorus sorption characteristics. It was found that the pond sediments had strong phosphorus sorption capacities. The phosphorus adsorption maximum (Smax) ranged from 228 to 974 mg x kg(-1), equilibrium phosphorus concentration (EPC0) ranged from 0.004 to 0.032mg L(-1). The mean degree of phosphorus saturation (DPS) of the pond sediments was only 9.5%. The situation of the ponds can influence the physico-chemical properties of sediments and their sorption characteristics. For Smax, Hill pond >Nonirrigation pond> Rice pond> River pond> Village pond. On the contrary, for EPC0, Hill pond< Nonirrigation pond< Rice pond < River pond < Village pond. The phosphorus adsorption maximum (Smax) was significantly correlated ly correlated with the KCl-extractable phosphorus (KCl-P, r2 = 0.83, p < 0.001). Multiple regression analysis shows that Fe(ox) and KCl-P are the key factors, which can influence phosphorus sorption of the pond sediments. PMID- 16212172 TI - [Chemical characteristics of nitrogen and phosphorus in the sediments of the typical bays in Dianchi Lake and calculation of their dredging layers]. AB - The sediments of Haidong bay and Macun bay in Dianchi Lakes' experimental area were studied. The sediment column was cut in every 3cm, and the following items are tested, such as total phosphorus, phosphorus species, which include labile phosphate, Fe,Al-P, Ca-P and residual-P, total nitrogen, nitrate nitrogen, nitrite nitrogen, ammonium nitrogen. The results showed that the loads of nitrogen and phosphorus of these two bays increased quickly in recent years. The Fe, Al-P was the largest component in all the P-forms and the content of ammonium nitrogen in these two bays less than that of other lakes. The layer of 9 to 12 centimeters of the two bays was polluted less than the rests. It indicated that these years more and more wastewater was inflow in these two bays. At the same time, the distribution of each testing items showed another low value between 24 to 39 centimeters. So considering feasibility and economic restriction, we can set the 24 - 39 centimeters depth as a dredging layer. PMID- 16212174 TI - [Optimal process combination for control of disinfection by-products and precursors]. AB - It is compared that the efficiency of control disinfection by-products and their precursors by sequential-chlorination disinfection and traditional free chlorination disinfection following different processes combination. Compared with traditional chlorination, sequeutial chlorination, i. e. short-term free chlorine plus chloramine disinfection can lower 35.8% - 77.0% of trihalomethane formation and 36.6% - 54.8% of haloacetic acids formation. Moreover, the poorer the influent quality is, the more advantages sequential chlorination disinfection have over free chlorination disinfection. The formation of trihalomethane and haloacetic acids by sequential chlorination following the simplest traditional process is even less than their formation by free chlorination following the most complex process, pre-ozonization plus traditional one and ozonization-active carbon process. The results show that ozonization-active carbon process and pre ozonization have better efficiency on control of disinfection by-products and their precursors. We recommend that water plants apply the sequential chlorination disinfection and ozonization-active carbon process. PMID- 16212173 TI - [Effects of plants on nitrogen/phosphorus removal in subsurface constructed wetlands]. AB - Effects of plants on nitrogen/phosphorus removal was studied in pilot-scale in subsurface constructed wetland, the main contents included nutrient uptake, effects of harvesting and roots on hydraulic condition. The result show that the amounts of nitrogen and phosphorous removed by plant harvesting is about 5% of the total removed nutrients in SFS wetlands. The best harvesting periods is between 9-10 month every year. Plant harvesting may induce fluctuation of outflow; aboveground biomass can stabilize micro-environmental of roots. The roots can also improve hydraulic condition of SFS system, decreasing dead area 5 % - 10 % and extending hydraulic retention time. PMID- 16212175 TI - [Relationship between algal growth and nutritious materials absorbability in the Three-Gorges Valley]. AB - According to measuring data of various relative pollution materials such as various nitrides, phosphides and chlorophyll along the Three-Gorges valley, and with introducing a special controlling function of new strong nonlinear coupling dynamics model, in which could used to describe complex behavior and process of green algal bloom to exist in slow flow, we studied algal growth mechanism and found some strong interactions and inherence rules when N and P concentration absorbated by algae body varied with the different N and P concentration in the valley. The ratio factor (wi/cx) of the nutritious absorption coefficient of algae wi to nutritious material concentration cx not only reveals the influence relation about nutrition content absorbated by the algae at different algal concentrations, but also characterizes the some correlations between algal growth and various effects of different nutritious materials absorbated by the algae. PMID- 16212176 TI - [Experiment study on real-time controlling rules of A/O nitrogen removal process]. AB - The aim of this work is operational optimization of an A/O process in the lab plant with synthetic wastewater for improving nitrogen removal efficiency. Ammonia control aims at maintaining the required concentration of ammonia in the effluent by manipulating the dissolved oxygen (DO) set point and aeration volumes. Nitrate control aims at the optimal use of the denitrification potential at any moment by continuously adjusting the internal recirculation flow or (and) external carbon addition flow in order to maintain a desired nitrate set point in the anoxic zone. The control strategies have been based on a hierarchical structure where a high level or supervisory control selects the set point of the low level or conventional controllers. Results indicated that it was possible to increase nitrogen removal efficiency, improve the effluent water quality, save energy, and reduce operating costs. PMID- 16212177 TI - [Full-scale two-phase anaerobic process treating traditional Chinese medicine wastewater]. AB - A navel two-phase anaerobic-aerobic treatment technology was developed to treat high concentration and refractory organic wastewater discharge from Harbin Second Traditional Chinese Medicine Factory. The function of two-phase anaerobic process in the whole bio-treatment system was analyzed. Now the status in bio-technique of treating wastewater from traditional Chinese medicine production was that all the influent COD concentration was under 5 000 mg/L and its quality was easy to be bio-treated. The full-scale experiment has changed this status. Full-scale experiment results showed that when influent COD concentration was at 7 000 - 40 000 mg/L and its BOD5 /COD was lower than 0.2, the volume-loading rate and COD removal rate of acidogenic phase, a continuous-flow stirred tank reactor, could reach 20- 30 kg/(m3 x d) and 47.1% respectively. At the same time, it only took the mathanogenic phase 27 days to set-up. The volume-loading rate and COD removal rate of mathanogenic phase, an up-flow anaerobic sludge blanket-anaerobic filter reactor, could reach 6.0 - 7.0 kg/(m3 d) and 94.06% respectively. It was also demonstrated the COD removal rate of two-phase anaerobic process was over than 93.0%, which offer endurance for the effluent from the whole treatment system reaching relative standard performed in China. PMID- 16212178 TI - [Preparation and characterization of a novel adsorbent made by cellulose acetate]. AB - A novel adsorbent made by cellulose acetate was developed. Preparation method, structure, and adsorbing characterization of the adsorbent were discussed. SEM results showed that the surface of round adsorbent was stable membrane of cellulose acetate, free from obvious cracks, holes, or other defects; while the cross section of the adsorbent was meshy and a lot of cavities were found. The adsorption results of 4 organochlorinated pesticides, such as Dieldrin, Aldrin, Endrin and Heptachlor, show that the adsorbent has higher efficient for organic pollutants, the adsorption rate is about 85% after 12 h. The adsorption rate is faster with the higher lgKow, and the removal efficiency of Heptachlor and Aldrin is up to 99% after 0.5h. The adsorbent can be used to remove persistent organic pollutants effectively. PMID- 16212179 TI - [Preparation of triolein-embedded CA membrane and its characteristics]. AB - Triolein is successfully embedded into cellulose acetate (CA) by phase inversion. This prepared flat membrane can effectively remove trace lipophilic organic pollutants from water. Structure of hybrid membrane is mainly observed by scanning electron microscope (SEM). Triolein dispersion by mechanical rabbling and ultrasound are investigated. Ultrasound can more effectively strengthen triolein dispersion than mechanical rabbling. Effect of casting membrane temperature (room temperature, 0 degrees C) shows low temperature can help to forming smaller triolein droplets. In addition, interaction between triolein and CA belongs to physical mixing by the observation of FT-IR, accordingly triolein structure is not changed and adsorptive capacity for persistent organic pollutants is not affected. Triolein in hexane is analyzed by fluorometric measure. The results show that triolein is completely embedded into membrane, so it is impossible that triolein leaks into water in the process of the adsorption. PMID- 16212180 TI - [Fractal characteristics and scale dependence of mature granular sludge in anaerobic baffled reactor (ABR)]. AB - Based on the SEM-images of mature granular sludge in ABR, the research process on fractal characteristics of these granules was built. The treatment and fractal dimensions calculation of the images were carried on through Photoshop, Newscan and Fips programs. The results showed that the boxing-counting dimensions of the mature granular sludge were more than 1.85, and the granules formed in the third and fifth compartments were more compact. The boundaries of the same granular sludge were fractal with the fractal dimensions of approximate 1.10, demonstrating the irregularity of the boundaries or surfaces, furthermore the surface of these granules formed in the first and fifth compartments was more irregular. It seemed that the compass dimensions of the granules boundary were a proof of irregularity of their surface. Moreover, there existed two scale scopes in calculating the compass dimensions of each granule, especially some of them with a range of one order of magnitude. A little discrepancy was appeared between box-counting and compass ways to calculating fractal dimensions. It will be important to build relationship between fractal dimensions with different scales and composition, structure, physico-chemical characteristics of granular sludge. PMID- 16212181 TI - [Improvement of the activity of activated sludge by low intensity ultrasound]. AB - Aerobic activated sludge from domestic wastewater treatment plant was used as an experimental material. Oxygen Uptake Rate(OUR) was detected as the index to indicate the changes of sludge activity induced by ultrasound(US) at 35kHz in 0 - 40 min with the US intensities of 0 - 1 .2 W/cm2. The best results were achieved at a US intensity of 0.3 W/cm2 and irradiation time of 10 min. Improper US intensity and irradiation time lead to a decrease of sludge activity. Therefore, the stimulating effects of US on the sludge activity could be enhanced at an optimized US intensity and irradiation time. In addition, changes of sludge activities (within 48 hours) after US irradiation at 0.3 W/cm2 in 10 min were examined. The activity of sludge took 8 hours after irradiation to reach peak level, and the activity was as double as that of the initial level after irradiation. In 24 hours after irradiation, the enhancement effects induced by ultrasound disappeared. Therefore, to improve the treatment efficiency of biological reactor, ultrasound with intensity of 0.3W/cm2 could be employed to irradiate part of the sludge in the biological treatment system for 10 minutes every 8 hours. It also presented the hypothetical explanation of the mechanism of biological activity enhancement stimulated by ultrasound. PMID- 16212182 TI - [Distribution and sources of polycyclic aromatic hydrocarbons in sediments from rivers of Pearl River Delta and its nearby South China Sea]. AB - Polycyclic aromatic hydrocarbons (PAHs) are measured in surface sediments from rivers and estuary of Pearl River Delta and its nearby South China Sea. Total PAH concentration varied from 255.9 - 16 670.3 ng/g and a moderate to low level compare to relevant areas worldwide. The order of PAHs concentration in sediments was: rivers of Pearl River Delta > estuary > South China Sea, and the most significant PAH contamination was at Guangzhou channel of Zhujiang river. A decrease trend for PAHs concentration with distance from estuary to open sea can be sees in South China Sea. Coal and biomass combustion is the major source of PAHs in nearshore of South China Sea, and petroleum combustion is the main source of pyrolytic PAHs in rivers and estuary of Pearl River Delta according to PAHs diagnostic ratios. Petroleum PAHs are revealed have a high contribution to PAHs in Xijiang River, estuary and some stations in Zhujiang River. A comparison of data from study in 1997 with data from present study indicates that there is no clear change in the PAH concentration over time but the source of PAHs in Pearl River Delta have been change from a main coal combustion to petroleum combustion and being reflect in the sediments in rivers and estuary of Pearl River Delta where there have high sedimentation rate. PMID- 16212183 TI - [Matrix-bound phosphine (PH3) distribution characteristics in the sediments of Jiaozhou Bay, China]. AB - The distribution of matrix-bound phosphine in sediments was investigated in some selective sea areas of Jiaozhou Bay on Sep.6, 2003. Results show that the peak concentration of phosphine reachs to 143.75 ng/kg (dry weight). Moreover, an obvious regularity of distribution is observed that phosphine concentration in coastal sediments is higher than that in offshore's, and concentration within the Bay is higher than that of outside Bay. Results also reveal that phosphine concentration in such seriously polluted areas as culture farm and estuary is significantly higher than that in other investigated places. On the other hand, the relationship between phosphine and its sedimentary environment suggest that the phosphine concentration and distribution are mainly affected by the organic phosphorous, the redox potential and the sediment composition. PMID- 16212184 TI - [Fate of matrix-bound phosphine during acidification with anaerobic bacteria]. AB - Matrix-bound phosphine of anaerobic granular sludge was studied during methanogenic and acidogenic stage of anaerobic reactors. Results showed that at methanogenic stage, the concentration of matrix-bound phosphine of anaerobic sludge decreased as the height of reactor increased. But during acidogenic stage, matrix-bound phosphine disappeared when pH of the solution declined to 4-5. Further anaerobic static experiments showed that increased acidification and lower pH value promoted the decrease of matrix-bound phosphine. After cultivation with 2 000 mg x L(-1) glucose synthetic wastewater for two days, the matrix-bound phosphine fell from 1.76 ng x kg(-1) to 0.09 ng x kg(-1); At the same time, the highest disappearing rate of phosphine was reached 0.84 ng x (kg x d)(-1). While the glucose synthetic wastewater was 400 mg x L(-1), the disappearing rate of phosphine was only 0.27 ng x (kg x d)(-1). With further cultivation under acidogenic stage, the phosphine decreasing rate slowed down. PMID- 16212185 TI - [Forming mycelium pellet and decolorization of dye wastewater under opening conditions]. AB - Removal of three water-soluble dyes through biosorption of four fungal strains was investigated. Some key factors affecting the formation of pellets and removal of dye under opening conditions was examined and the possibility of repeated inoculation of pellets was evaluated. The results show that removal of acid brilliant red B by 4 strains was almost 100%; the removal of all three tested dyes by Trichoderma sp. was more than 99%. Trichoderma sp. formed pellets well under opening conditions when its fragmentized mycelium obtained by 3 min grinding pellets at high concentration was inoculated, as 100% removal of acid brilliant red B was achieved by biosorption of newly growing pellets. Repeated inoculation of pellets accelerated dye removal, time for 100% decolorization reduced from first inoculation 72h to third inoculation 12h. PMID- 16212186 TI - [Improvement of heavy metal removing strain by protoplast combination mutant]. AB - Ultraviolet and HNO2 were selected as the mutagens to perform the single factor and multi-factor induction mutation towards Candida utilis CR-001. Six mutant strains which possessed high heavy metal removal efficiency and high resistance to Cr6+ were obtained through combined induction with UV and HNO2. After they were subcultured for 10 generations, the diameter of bacteriostatic circle of CRC2811-1 and CRC7-2 was reduced to 1.7 mm and 1.2 mm respectively, while the Cr6+ removal efficiency of CRC2811-1 was increased from 80.2% to 95.2%, and that of CRC7-2 was from 81.2% to 94.7%. The stability of the other 4 mutant strains was rather stable. Furthermore, precipitation of chromium outside or inside the cell was studied by using combined technique of scanning electron microscopy(SEM), transmission electron microscopy (TEM), and atomic force microscopy(AFM), and the mechanism of chromium removal improvement by the mutant strains was discussed. PMID- 16212187 TI - [Enantioselective degradation of 2, 4-dichlorprop methyl ester by sediment bacteria]. AB - Enantioselective Degradation of 2,4-Dichlorprop methyl ester was studied by sediment bacteria. One DCPPM-degrading bacteria stain was isolated from contaminated sediments. The EF values were determined by chiral GC. The enantioselectivity of DCPPM biodegradation in aquatic phases were evaluated. Incubation with DCPPM-degrading bacteria show that the R enantiomer was preferentially degraded over the corresponding S enantiomer of DCPPM. The EF values show a much higher deviation. It indicate that the microbial degradation of DCPPM was enantioselectivety. PMID- 16212188 TI - [Biodegradation of butylbenzyl phthalate by acclimated activated sludge]. AB - The results of degradation of butyl benzyl phthalate(BBP) by activated sludge demonstrate that the activated sludge is a high efficiency flora for degrading BBP, which can degrade BBP quickly and more than 95 % BBP is degraded in one day. The degradation of BBP can be described by the first-order dynamics model, the concentration in range of 100,300,500 mg x L(-1), the dynamics equation of degradation respectively is Inc = - 0.0699x + 5.6171, nc = 0.0683x + 8.4083, and Inc = - 0.0583x + 7.2806, t1/2 is 9.92, 10.15, 11.89 h, the constant of degrading rate kb is 0.0699, 0.0683 and 0.0583 d(-1). The constant of degrading rate decreases as the concentration of BBP increases, which shows that BBP of high concentration inhibits its biodegradation. In addition, two degraded productions, monobutyl phthalate and methanoyal phthalate, are identified from the degradation of BBP with HPLC-MSn. The route of degradation is that BBP produces mono-ester firstly, and then the product turns into phthalic acid, finally it oxidizes into CO2 and H2O. PMID- 16212189 TI - [New method for spatial structure quantification of biofilm with GFP tagged bacteria]. AB - Green fluorescent protein (GFP) gene was applied to mark E. coli JM109 by the method of CaCl2 transformation. The GFP-tagged cells was inoculated into the LB medium containing 50 microg/mL of Ampicillin to develop biofilm on the surface of lava by mix-culture with lava particles in flask under 37 degrees C, 120 r/min for 16 h. The fluorescent images stack of different layer at specific area (250 microm x 250 microm) on lava covered with biofilm was obtained by confocal scanning laser microscopy. The quantifying parameters of biofilm spatial structure could be acquired after the image information was calculated by COMSTAT program, i. e., the average thickness of specific area of biofilm after 16 h was 0.120 844 microm, the maximum thickness was 10.5 microm, the volume was 0.136 986 microm3 /microm2, the surface area was 21 338.1 microm2, and the surface area per volume biofilm was 3.36 854 microm2 /microm3. This method might be expanded to reflect the spatial structure of biofilm with any other GFP tagged bacteria. PMID- 16212190 TI - [Metabolic properties of the microbial community in the biofilters using biolog microplates]. AB - It is very important to know the structure and metabolic function of the microbial community in a bioreactor in order to improve its performance. In this study, two biofilters, packed with wood chips and granular activated carbons respectively, were operated for 160 days to treat toluene gas. The metabolic profiles of the microbial communities in the biofilters were monitored using Biolog microplates periodically during the experiments. The metabolic activities of the microorganisms in both biofilters were observed to decrease during long term operation. According to the results of principle components analysis, the metabolic profiles of the microbial communities did not change much in the former period of the operation, but they changed in the inlet layers on day 103 and changed throughout the filter beds on day 160. The variation of the metabolic profiles in both biofilters showed little difference, which suggested that the packing media had little effect on them during long-term operation. Among the 95 carbon sources in Biolog microplate, carboxylic acids and amino acids were much easier to be utilized by the microorganisms in the biofilters than the other carbon PMID- 16212191 TI - [Microbial community structure analyzed by single-strand conformation polymorphism technique in sulfate-reducing reactor]. AB - Analyses of microbial community structure and the relationships between Sulfate Reducing Bacteria (SRBs) and Acidogenic Bacteria (ABs) in a completely stirred sulfate-reducing reactor were carried out by modified polymerase chain reaction single-stranded conformation polymorphism (PCR-SSCP) targeted eubacterial 16S ribosomal RNA gene. A total of 13 bands were obtained and 6 of them (A1, A3, A4, A5, A9, A10) were sequenced. The sequences are similar to Leuconostoc mesenteroides (GenBank Access No. AY453065), some uncultured bacteria (AJ318147, AF227834, AJ576427), Ethanologenbacterium (AY434722), Clostridiaceae (AB084627), etc. In order to investigate the SRBs in the reactor,the active sludge was cultured on SRB-selected media and also did SSCP with the compound cultured bacteria. Two new bands appeared, one similar to Bacteroidetes (AB074606) and another similar to Desulfovibrio (Y12254, U42221). The experimental results indicate that the proportion of SRBs in the reactor is probably less than 1.5 percent. But the few SRBs play a very important role during the course of sulfate reduction via cooperation with acidogenic PMID- 16212192 TI - [Uptake HTO from seawater by seashell and formative dynamics of bound tritium in seashell]. AB - Uptake HTO from seawater by five kinds seashell and formative dynamics of bound tritium in organ of seashell are studied by using the isotope-tracer techniques in order to get a better understanding of the marine environmental behavior of HTO. The results show that the tritium in seashell exists in free water tritium and bound tritium form. The free water tritium is main form; its content observed was more than 97.4% of total tritium. The bound tritium content was very lower, only being 0.4% - 2.6% in total tritium. The speed of HTO untaken by seashell is very rapid, specific activity of HTO in three kinds seashell had reached their maximum only 2 h after treatment. The bound tritium in seashell increases slowly with time. The results of analysis of concentration factor (CF) values indicate that HTO would not concentrate in the seashell. PMID- 16212193 TI - [Application of biosurfactant in composting of agricultural waste]. AB - The biosurfactant of rhamonolipid (Rh) obtained from Pseudomonas aeruginosa was used in co-composting of rice straw and wheat bran, the efficiency of the promoter was studied under temperature control during the first stage of composting. Composting was done in an aerobic static bed of composting during which moisture content of mixture materials was controlled between 60% 70%, and the control of aeration adopted the timer by turns of 0.18m3/h for 20 minutes and pause for 40 minutes. Changes were studied including pH, organic matter, dissolved organic carbon (DOC), the biomass of microorganism, hemicellulase activity, carboxymethyl cellulose enzyme (CMCase) activity, hemi-cellulose and cellulose contents. The experimental results show that the degradation rate of organic matter is 13.4% higher than that of the control, and the average of DOC is improved by 2.2g/kg. The degradation contents of hemi-cellulose and cellulose are increased by 5.7%, 10.7% compared with the control. This indicate that the addition of Rh can improve microenvironment, strengthen polymers hydration, accelerate the co-composting process and improve the quality of composting production. PMID- 16212194 TI - [Construction and function of a high-efficient complex microbial system to degrade cellulose and lindane in compost]. AB - A complex microbial system capable of degrading cellulose and lindane with high efficiency was isolated from four compost heaps. It was selected and domesticated through two methods and by combination of different microbial communities. The results show that the complex microbial system can decompose filter paper, absorbent cotton, rice straw powder and sawdust effectively, especially has high degrading activity for the materials with higher native cellulose such as filter paper and absorbent cotton. As for both of them, the CMC saccharification activity is more than 40U and the degradation efficiency is more than 95% on the 5th day of inoculation. The complex microbial system can also keep a higher degrading capability in a wider range of pH. Filter paper and lindane can be degraded effectively by the complex microbial system during pH 7.0 - 9.0, and the degradation rates are more than 90 % and 45% respectively. Under pH 6.0 - 9.0, there is a good consistency between the degradation of Lindane and the decomposition of filter paper. PMID- 16212195 TI - [Heavy metal stabilization in municipal solid waste incineration fly ash using soluble phosphate]. AB - The physico-chemical characteristics of municipal solid waste incineration fly ashes was analyzed. It indicated that the main elements of fly ashes are Ca, Cl, K, S and Si, and many heavy metals such as Pb, Zn, Cu, Mn and Cr can be found in fly ashes, and the heavy metal leaching toxicity such as Pb, Cu, Zn and Cd is much higher than the standards. A new kind of stabilization agent-soluble phosphate is chosen to treat with fly ash, different influence factors including agent dosage, curing time and pH are taken into consideration to test its stabilization for the stabilized fly ash. The experimental results indicate that fly ash treated with soluble phosphate can have excellent stabilization effect, and the leaching toxicity for Pb, Cd and Zn can be reduced by 97.5%, 91.6% and 95.5% at a phosphate dosage of 3%; curing time can not influence its stabilization; and, stabilized fly ash using soluble phosphate can keep long-term stabilization at a broad pH value. PMID- 16212196 TI - [Modification of MSWI fly ash by using anionic chelating surfactant]. AB - This paper aims at the reutilization and stabilization of municipal solid waste incineration (MSWI) fly ash to produce a high value added product. The effects of anionic chelating surfactants on the surface modification and fixing capacity of MSWI fly ash were primarily explored. Based on the indexes of active ratio and analysis of IR-spectrometer, the active ratio of ACS1 modified fly ash can be found higher than 95% under the condition of surfactant dosage at 7.0mL/100g fly ash, modification time at 15min and temperature at 75 degrees C . Moreover, Anionic chelating surfactant shows a strong fixing capacity for heavy metals by the United States Environmental Protection Agency's Toxicity Characteristic Leaching Procedure (TCLP). PMID- 16212197 TI - [Ecological footprint in building green university]. AB - Ecological footprint is one of the best indexes to evaluate the level of green university. The approaches of ecological footprint are made up of compound approach and component approach. This paper introduces the basic principle and algorithm of the componential approach of ecological footprint, taking Northeastern University as an example and using this approach in the research of campus. Result show that the ecological footprint of Northeastern University 2003 was 24 787hm2, needing the productive land of ecology about 25 000hm2 support all kinds of consumption of the school and absorb the offal. The ecological efficiency of the school was 0.94cap/hm2. In the ecological footprints, the energy's footprints is the largest, it accounted for more than 2/3 of the total footprints, the next are food consumption and solid rubbish. PMID- 16212198 TI - [Structure and ecological environmental efficiency of urban forestation]. AB - It is proposed that a new pattern of forestation, arbor-shrub-lawn composite construction, based on analyzing the form and the actual state of the urban forestation. The arbor-shrub-lawn is superior by comparing with the urban lawn as for ecological environmental function including landscape. It is suggested that forestation's ecological saving efficiency and management, maintaining and reducing cost should be thought over simultaneously. PMID- 16212199 TI - Road safety: the potholes of neglect. PMID- 16212201 TI - Patterns of antenatal care in low-versus high-risk pregnancies in Lebanon. AB - We studied patterns of antenatal care in low- versus high-risk pregnancies in Lebanon comparing 538 women after delivery in urban Beirut with rural Baka'a. Most women had 9 antenatal care visits with an obstetrician, starting in the first trimester. Care for high-risk and low-risk pregnancies was similar in terms of type of provider, number of visits and timing of first visit. More high-risk women had advice about special diets, supplements and laboratory tests. Maternal and fetal outcomes showed that, controlling for area and pregnancy risk, more antenatal visits were associated with fewer preterm deliveries, more caesarean sections and fewer cases of postpartum depression. Overall, differences between risk groups were small. PMID- 16212200 TI - Contraception continuation rates and reasons for discontinuation in Zahedan, Islamic Republic of Iran. AB - We evaluated contraception continuation rates and discontinuation reasons in Zahedan among 1741 women from 1998-2000. By Kaplan-Meier technique continuation rates were 92% for low dose combined hormonal oral contraceptives (OC), 86% for a levonorgestrel-releasing implant, 82% for intrauterine devices (IUD) and 53% for medroxyprogesterone acetate at the first year. After 3 years, continuation was 78% for levonorgestrel implant, 70% for OC, 60% for IUD and 44% for medroxyprogesterone acetate. The commonest reason for discontinuing OC and medroxyprogesterone acetate was changing method; for IUD and levonorgestrel releasing implant, the commonest reason was side-effects. By Cox regression model, continuation rate and contraceptive type were significantly related to health centre. PMID- 16212202 TI - Ceftriaxone versus ampicillin/cloxacillin as antibiotic prophylaxis in elective caesarean section. AB - We carried out a prospective, randomized, controlled clinical trial to evaluate the clinical efficacy of ceftriaxone and ampicillin/cloxacillin prophylaxis in decreasing the frequency of post-caesarean section infection-related morbidity. Two hundred patients randomly received either ceftriaxone (single dose) or ampicillin/cloxacillin (3 doses) intravenously at induction of anaesthesia. There was no statistical difference in incidence of endometritis (P = 0.34), wound infection (P = 0.44), or other febrile morbidity (P = 0.5). Eleven babies had a low Apgar score (< 8) in the ceftriaxone group and 13 in the ampicillin/cloxacillin group (P = 0.82). There were 2 perinatal deaths in each group. One dose of ceftriaxone was as effective as ampicillin/ cloxacillin in preventing post-caesarean section complications and is easier to administer. PMID- 16212203 TI - Effect of exclusive breastfeeding and complementary feeding on infant growth and morbidity. AB - A cohort study was conducted in the Islamic Republic of Iran between January 1997 and February 1998 to compare the growth and morbidity of 100 infants who were exclusively breastfed for 6 months and 100 who received breast milk and complementary foods between 4-6 months. Infants' feeding pattern, weight and height were assessed and recorded. There were no significant differences in infants' weight and height gain between 4 and 6 months. The rate of diarrhoea between ages 4 and 6 months was significantly lower in exclusively breastfed infants than in complementary food-fed infants (11% versus 27%) and respiratory infections were also lower (23% versus 35%). We conclude that exclusive breastfeeding is superior at least until an infant is 6 months of age. PMID- 16212204 TI - Socioeconomic predictors of unconstrained child growth in Muscat, Oman. AB - We conducted a study to identify socioeconomic factors associated with unconstrained growth among preschoolers in Muscat, Oman. A sample of children born in 1995 and aged 28-43 months was drawn from the Child Health Registers of 2 health care centres. Sociodemographic data were collected by oral interview and maternal and child anthropometry measured. Regression analysis was used to identify socioeconomic indicators and cut-offs associated with unconstrained growth. Children from households with a monthly income > or = 800 Omani rials and mother's education > or = 4 years attained height-for-age levels comparable to the current international growth reference. By screening using this criterion, we could obtain a suitable sample for the World Health Organization Multicentre Growth Reference Study in Muscat. PMID- 16212205 TI - Cost-effectiveness of audiometric screening of first-year preparatory pupils in Dhofar Region, Oman. AB - Audiometric screening was conducted in Dhofar region to study the magnitude of ear problems and cost-effectiveness of screening first-year preparatory-school children in Oman. None of the 1894 pupils had otitis media with effusion or sensory neuronal hearing loss. Six children (0.32%) had impacted wax, 4 (0.21%) chronic suppurative otitis media and 2 (0.11%) dry perforation of eardrum. In all, 14 children (0.74%) with suspected hearing impairment were referred to a specialist but only 2 attended. Physicians and nurses spent 8-10 minutes for ear examination per child for a yield of less than 1%. The screening expenditure was US$ 5 per pupil. As the prevalence of serious ear conditions was low, we conclude that expanding the audiometric screening of schoolchildren to first-year preparatory pupils is not cost-effective. PMID- 16212207 TI - Influenza surveillance in the Islamic Republic of Iran from 1991 to 2001. AB - To better understand the annual distribution of influenza virus in our country, we isolated and typed 45 viruses from 1043 patients with acute respiratory illnesses in a 10-year study conducted by the National Influenza Centre of the Islamic Republic of Iran. The seasonal distribution of influenza typically ran from November to April. Type A influenza was most common during the winters of 1991-92, 1997-98 and 2000-01 and type B influenza was most common during 1992-5 and 1996-97. Both type A and type B viruses circulated in 1995-96 and 1998-2000. A serological survey based on haemagglutination inhibition test confirmed our findings. The annual pattern of strains isolated was similar to the worldwide pattern during the same interval. PMID- 16212206 TI - Efficacy of sulfadoxin pyrimethamine for uncomplicated Plasmodium falciparum malaria in a small sample of Sudanese children. AB - A prospective clinical trial was carried out to determine in vivo efficacy of sulfadoxine/pyrimethamine for the treatment of uncomplicated Plasmodium falciparum malaria in children in New Halfa. Forty patients were enrolled; 31 completed the 28-day follow-up. Six (19.4%) patients showed recurrence of parasitaemia during follow-up, while the rest (80.6%) cleared the parasites and responded fully to treatment. All the failures were late treatment failures. Parasite genotyping showed that 1 (16.7%) of the 6 cases of late parasitaemia was due to reinfection while the rest (83.4%) were due to true recrudescence. During the follow-up period 22.6% of patients showed gametocytaemia. The high level of treatment failure as well as gametocytaemia necessitates the introduction of artesunate in this combination therapy. PMID- 16212208 TI - Antimicrobial resistance and plasmid profiles of urinary Escherichia coli isolates from Jordanian patients. AB - We investigated antimicrobial resistance patterns and plasmid profiles of uropathogenic Escherichia coli isolates from inpatients and outpatients at Jordan University Hospital in 2000 and 2001. E. coli accounted for 32.4% and 37.4% of all isolates respectively. The lowest susceptibility was for ampicillin (11%), cotrimoxazole (23%) and tetracycline (26%). The relative incidence of resistant isolates of E. coli to nalidixic acid, gentamicin, norfloxacin, cefuroxime and nitrofurantoin was significantly greater for inpatients than for outpatients (P< 0.05). A large, transferable R-plasmid of 28 kb was found in most E. coli isolates (67%) that were resistant to at least ampicillin, cotrimoxazole and tetracycline. This R-plasmid reservoir may contribute to the spread of multiple antibiotic resistance in our Region. PMID- 16212209 TI - Evaluation of the COBAS AMPLICOR MTB test for the detection of Mycobacterium tuberculosis complex. AB - We evaluated the COBAS AMPLICOR polymerase chain reaction (PCR) based test for the detection of Mycobacterium tuberculosis complex in 866 respiratory and non respiratory samples. Acid-fast staining and culture on Lowenstein-Jensen medium were also performed on all samples. Of the 866 samples tested, 87 (10.0%) were PCR-positive compared to 94 (10.9%) culture positive. There were no false positive results but 7 PCR-negative, culture-positive samples were, considered false negatives after reviewing medical records of patients. A PCR inhibitory rate of 2.0% (17/866) was observed in respiratory samples only. Sensitivity, specificity, and positive and negative predictive values for this test were 92.5%, 100%, 100% and 99.1% respectively. This test is a valuable diagnostic tool for today's mycobacteriology laboratory. PMID- 16212210 TI - Molecular evidence of mixed P. vivax and P. falciparum infections in northern Islamic Republic of Iran. AB - This study compared basic microscopy with molecular detection of Plasmodium species. According to thick-film microscopy, 100% of 142 malaria cases in Pars Abad, Ardebil province, were infected with a single species, P vivax. However, nested polymerase chain reaction (PCR) detected mixed species infections of both P. vivax and P. falciparum in 7.0%. In Mazanderan province, 2/20 blood films were diagnosed with only P. falciparum and 18/20 with only P. vivax. However, nested PCR detected 17/20, 2/20 and 1/20 with P. vivax only, P. falciparum only and mixed species respectively. The unexpected presence of P. falciparum urges prompt investigation and immediate treatment of malaria cases in this region. PMID- 16212211 TI - [Prevalence of infection by intestinal parasites in north Lebanon: 1997-2001]. AB - We determined the prevalence of intestinal parasites in the north of Lebanon between 1997 and 2001. We analysed the parasitology records of 17126 patients and evidence of parasitic infections was found in 5 713 (33.35%) cases. There was no significant difference in prevalence for males or females for any of the parasites. The most prevalent parasites were Entamoeba coli (38.45%), Ascaris lumbricoides (37.14%), Giardia lamblia (15.39%), Ent. histolytica (4.57%) and Taenia sp. (3.3%). A comparison between our data and results of previous studies in Lebanon in 1937, 1939, 1956, 1967 and 1993 showed an increase in the prevalence of A. lumbricoides and G. lamblia in the period 1997-2001, with less marked changes in the prevalence of the other parasites. PMID- 16212212 TI - A retrospective hospital study of human cystic echinococcosis in Egypt. AB - We performed a retrospective study to determine annual clinical incidence of human cystic echinococcosis (CE) in 14 Egyptian hospitals between January 1997 and December 1999. From 492 353 records examined, 133 (0.027%) new human CE cases were recorded. Of these, 50 (37.6%) were from Alexandria and Matrouh hospitals, 33 (24.8%) from Giza Chest Hospital and 50 from other regions. Matrouh governorate had the highest annual clinical incidence (1.34-2.60 per 100 000) followed by Giza governorate (0.80-1.16 per 100 000). About a third of those affected were aged < or = 20 years. Liver and lungs were the organs most affected. Although human CE is of low endemicity in Egypt, it may-represent a public health concern in Matrouh and Giza governorates. PMID- 16212213 TI - Inpatient satisfaction with physician services at King Khalid University Hospital, Riyadh, Saudi Arabia. AB - The satisfaction of 400 inpatients with physician services at King Khalid University Hospital, Riyadh was evaluated. Patient characteristics and ward of admission were collected and a questionnaire based on the standardized Likert scale was used. The highest mean satisfaction score was for admission and the lowest for communication. Among service items, the highest mean score was for physicians enquiring about patient conditions and opinions when planning care and the lowest for physicians asking for opinions about care quality and problems. Female and less educated patients were more satisfied with their care than male and educated patients. Male surgical and medical ward patients were the most dissatisfied with physicians'services. These findings offer hospital management information about shortcomings requiring remedial intervention. PMID- 16212214 TI - Primary health care services utilization and satisfaction among the elderly in Asir region, Saudi Arabia. AB - Use of primary health care (PHC) services and satisfaction among elderly people (60 + years) in Asir was studied in 26 PHC centres. They visited PHC centres significantly less often than younger adults but they were referred significantly more often to secondary and tertiary care and for more laboratory tests. A random sample of 253 elderly people attending the centres was interviewed about accessibility, continuity, humaneness, informativeness and thoroughness of care. Overall, 79.0% were satisfied with the services provided. The leading 3 items of dissatisfaction were: not enough audiovisual means for health education (65.1%), long time spent in the centre (46.4%), and not enough specialty clinics (42.5%). PMID- 16212215 TI - Performance of health providers in primary health care services in Jordan. AB - The study analysed time utilization by a sample of 111 physicians, nurses and midwives in 62 primary health care facilities in Jordan. The providers' activity over each shift was coded at 3-minute intervals using an activity sampling technique and patient-provider contact times were recorded. Overall, health providers spent the nearly half their time (48.7%) as down time' (waiting, breaks and non-work related activities), with 29.1% as clinical activities and 22.1% as non-clinical work-related activities. Physicians had higher clinical and down times than nurses and midwives; waiting for patients accounted for half the down time. The mean physician-patient contact time was 3.08 minutes. An appointment system is recommended to reduce down times for health providers and increase consultation times. PMID- 16212216 TI - Reducing health care costs by rationalizing staffing in primary care settings. AB - Jordan spends around 9% of its GDP on health care services, a high figure compared with similar developing countries. This study assessed staffing patterns in relation to Ministry of Health expenditures in a nationally representative sample of 97 primary care facilities. The economic costs of primary care facilities amounted to Jordanian dinar (JD) 42.3 million. Personnel costs consumed 53.8% of recurrent costs and in monetary terms the amount of down time (time not being used effectively) amounted to JD 9.7 million (about US$ 13.7 million). The Ministry should consider changing the functioning of its primary care facilities to obtain a more cost-effective use of staff time. PMID- 16212217 TI - Using the job characteristics model to compare patient care assignment methods of nurses. AB - The aim of this study was to use Hackman and Oldman's job characteristics model to compare 2 methods of patient care assignment as perceived by nurses in 12 inpatient units of Alexandria Main University Hospital. The job diagnostic survey was used to determine nurses' perceptions toward the components of the model in relation to their performance in utilizing the case and functional methods of patient care assignment. The jobs of intensive care unit nurses who utilized the case method were more enriched than those who utilized the functional method of assignment in the general care units, in terms of their perception scores towards all parts of the model (core job characteristics, critical psychological states, affective and personal outcomes, context satisfaction and individual growth-need strength). PMID- 16212218 TI - Evaluation of availability, accessibility and prescribing pattern of medicines in the Islamic Republic of Iran. AB - Prescribing, dispensing, availability and affordability of drugs were evaluated in 100 primary health care centres in 5 provinces of the Islamic Republic of Iran using WHO indicators. On average, 92% of the 12 essential drugs monitored were available in the health centre pharmacies and 95% of the drugs prescribed by the physician were dispensed by the health centre pharmacy. The stock-out duration was less than 1 month on average. A complete treatment for pneumonia cost only 2% of the lowest weekly government salary. The national average number of drugs per prescription was 3.4. Prescription of antibiotics and injectable drugs was very high (58% and 41% respectively). Although availability and affordability of essential drugs is good in this country, rational use of drugs needs to be emphasized. PMID- 16212219 TI - Prevalence of skin diseases among students with disabilities in Mansoura, Egypt. AB - Children with disabilities may be particularly susceptible to skin disorders, therefore the aim of this study was to estimate the prevalence of skin disease among such children in Mansoura, Egypt. A total of 636 students with disabilities (76 blind, 446 deaf-mute and 114 mentally retarded) and 720 sex and age matched students (control) who did not have these disabilities were given a thorough dermatological examination. We found 89.5% of blind students, 99.3% of deaf students and 100% of mentally retarded students had 1 or more skin diseases (both infectious and non-infectious) in comparison to 24.2% of the control group. Strict hygienic measures, periodic skin examination and health education of persons caring for students with disabilities are recommended. PMID- 16212220 TI - The presence of dental fluorosis in the permanent dentition in Doha. AB - The main purpose of this study is to draw attention to the presence and the severity of dental fluorosis in Doha (Qatar) through the high level of registered cases of fluorosis found among the groups examined. Of 4800 people aged 8-50 years, 2654 (55.29%) had dental fluorosis of some degree. This problem urgently requires more studies throughout the country to provide an accurate assessment. PMID- 16212221 TI - [Causes of drinking-water contamination in rain-fed cisterns in three villages in Ramallah and Al-Bireh District, Palestine]. AB - We studied the biological characteristics of drinking-water in three villages in Ramallah and al-Bireh district, by testing the total coliforms. Water samples were collected from rain-fed cisterns between October and November 2001. The results show that 87% of tested samples of drinking-water were highly contaminated and in need of coagulation, filtration and disinfection based on the World Health Organization guidelines for drinking-water, and 10.5% had low contamination and were in need of treatment by disinfection only. Only 2.5% of the tested samples were not contaminated and were suitable for drinking without treatment. The main cause of drinking-water con tamination was the presence of cesspits, wastewater and solid waste dumping sites near the cisterns. PMID- 16212222 TI - Normal uterine size in women of reproductive age in northern Islamic Republic of Iran. AB - To determine the efficacy of ultrasonographic assessment of uterus size in women of reproductive age, we conducted a cross-sectional analytic study of 231 women aged 15-45 years in Babol, northern Islamic Republic of Iran. Mean uterus size was 86.6 mm x 49.6 mm x 40.6 mm overall, 72.8 mm x 42.8 mm x 32.4 mm for nulliparous women and 90.8 mm x 51.7 mm x 43.0 mm for multiparous women. Mean age was 31.7 +/- 9.6 years and mean body mass index (BMI) was 24.7 +/- 4.0 kg/m2. Uterus size was significantly associated with parity and age; but not with BMI. Our findings show a greater mean uterus size than reported by others. Ultrasonographic measurement of uterus size is valuable for predicting pathologies associated with abnormal uterine size. PMID- 16212223 TI - Postage stamps as a health promotion tool in the Nepalese community. AB - Over the last 15 years the government of Nepal has issued postage stamps as a way of raising awareness of health issues in the general population and especially in remote communities. The topics covered by 8 different stamps are good child health care practices, combating drug abuse, hazards of smoking, prevention of AIDS, prevention of blindness due to cataract, fighting cancer and rehabilitation of disabled people. PMID- 16212224 TI - Successful outcome of pregnancy in a case of aplastic anaemia in Jordan. PMID- 16212225 TI - Managing a patient with thrombotic thrombocytopenic purpura requiring large amounts of blood in Oman. PMID- 16212226 TI - Pseudotumour due to retained surgical sponge (gossypiboma). PMID- 16212227 TI - Pharmacological action of Panax ginseng on the behavioral toxicities induced by psychotropic agents. AB - Morphine-induced analgesia has been shown to be antagonized by ginseng total saponins (GTS), which also inhibit the development of analgesic tolerance to and physical dependence on morphine. GTS is involved in both of these processes by inhibiting morphine-6-dehydrogenase, which catalyzes the synthesis of morphinone from morphine, and by increasing the level of hepatic glutathione, which participates in the toxicity response. Thus, the dual actions of ginseng are associated with the detoxification of morphine. In addition, the inhibitory or facilitated effects of GTS on electrically evoked contractions in guinea pig ileum (mu-receptors) and mouse vas deferens (delta-receptors) are not mediated through opioid receptors, suggesting the involvement of non-opioid mechanisms. GTS also attenuates hyperactivity, reverse tolerance (behavioral sensitization), and conditioned place preference induced by psychotropic agents, such as methamphetamine, cocaine, and morphine. These effects of GTS may be attributed to complex pharmacological actions between dopamine receptors and a serotonergic/adenosine A2A/ delta-opioid receptor complex. Ginsenosides also attenuate the morphine-induced cAMP signaling pathway. Together, the results suggest that GTS may be useful in the prevention and therapy of the behavioral side effects induced by psychotropic agents. PMID- 16212228 TI - Synthesis of new 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine derivatives with an incorporated thiazolidinone moiety and testing their possible serine protease and cercarial elastase inhibitory effects with a possible prospective to block penetration of Schistosoma mansoni cercariae into the mice skin. AB - 5-Substituted 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine were synthesized by interaction of 4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-sulfonylhydrazide with some aldehydes to give the corresponding Schiff-bases, which after cyclization gave corresponding thiazolidinones. For some of the thiazolidinones, Mannich bases reaction was carried out. All the derivatives were tested for their possible inhibitory effect on Schistosoma mansoni cercarial elastase (CE). Only, N-(4-methylbenzyledine)-4-oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine-5 sulfonylhydrazide was found to have potent inhibitory effect on the CE activity with IC50 = 264 microM. Upon its use as a paint for mice tails before infection with S. mansoni cercariae, the compound formulated in jojoba oil caused a significant reduction (93%; P-value = 0.0002) in the worm burden. IgG & IgM in mice sera were measured by using several S. mansoni antigens by ELISA. Sera from treated infected mice (TIM) 2, 4, and 6 weeks (W) post infection (PI) showed 1.2 folds lower, 1.2 folds higher, 1.7 folds lower IgM reactivity against soluble cercarial antigenic preparation (CAP), respectively, when compared with sera collected from infected untreated mice (IUM). Sera from TIM 2, 4, and 6WPI showed 1.3, 1.6, and 1.7 folds higher IgG reactivity, respectively against CAP than the IgG reactivity from IUM. Sera from TIM 2, 4 and 6WPI showed 1.5, 1.2 folds lower and 1.4 folds higher IgM reactivity, respectively against soluble worm antigenic preparation (SWAP) when compared with sera collected from IUM. Sera from TIM 2, 4, and 6WPI showed 1.4, 1 folds lower and 1 fold higher IgG reactivity, respectivley to SWAP when compared with sera from IUM. Sera from TIM 2, 4, and 6WPI had generaly lower IgM and IgG reactivities against soluble egg antigen (SEA) when compared with sera from IUM. PMID- 16212229 TI - Synthesis and antiinflammatory activity of 1,5-diarylimidazoles. AB - A number of 1,5-diarylimidazoles has been synthesized and evaluated for their inhibitory activities of COX-2 catalyzed PGE2 production. 1,5-Diarylimidazoles were obtained from imimes and p-toluenesulfonylmethyl cyanide (TosMIC). Imines were prepared from commercially available amines and aldehydes. Among the compounds tested, 1-(2,4-difluorophenyl)-5-(4-methylsulfonylphenyl)imidazole (2r) showed strong inhibitory activity, however, most diarylimidazoles exhibited little to low inhibitory activities against COX-2 catalyzed PGE2 production. PMID- 16212230 TI - An efficient synthesis of risperidone via stille reaction: antipsychotic, 5-HT2, and dopamine-D2-antagonist. AB - Risperidone has been reported to have neuroleptic activity. In this study, risperidone was synthesized using a new method involving a stille reaction, in which 2-methyl-3-vinyl-6,7,8,9-tetrahydropyrido[1,2-a]pyrimidin-4-one was synthesized (5). The chemical synthesis process was found to be simple and produced risperidone in a high yield. In addition, can be easily scaled up for large scale synthesis. PMID- 16212231 TI - A new furofuran lignan with antioxidant and antiseizure activities from the leaves of Petasites japonicus. AB - A new furofuran lignan (1) was isolated from the n-butanol fraction of the methanolic extract of the leaves of Petasites japonicus (Sieb. et Zucc.) Maxim. (Compositae). The structure of compound 1 was determined to be 2alpha-(4'-hydroxy 3'-methoxyphenyl)-6alpha-(4"-hydroxy-3"-methoxyphenyl)-8alpha-hydroxy-3,7 dioxabicyclo[3.3.0octane 4'-O-(beta-D-glucopyranoside) by spectroscopic methods including 2D-NMR. In further studies, it was found that the compound 1 expressed an antioxidant activity in DPPH radical scavenging assay, and moreover, ameliorated the seizure in kainic acid-treated mice. PMID- 16212232 TI - Antidiabetic stilbene and anthraquinone derivatives from Rheum undulatum. AB - The antidiabetic-activity-guided fractionation and isolation of the 80% EtOH extracts obtained from cultivated Korean Rhubarb rhizomes (Rheum undulatum, Polygonaceae) led to the isolation and characterization of one stilbene, desoxyrhapontigenin (1) and two anthraquinones, emodin (2) and chrysophanol (3). Their structures were established by chemical and spectroscopic methods. Compounds 1, 2, and 3 inhibited postprandial hyperglycemia by 35.8, 29.5, 42.3%, respectively. PMID- 16212233 TI - Antimicrobial property of (+)-lyoniresinol-3alpha-O-beta-D-glucopyranoside isolated from the root bark of Lycium chinense Miller against human pathogenic microorganisms. AB - (+)-Lyoniresinol-3alpha-O-beta-D-glucopyranoside (1) was isolated from an ethyl acetate extract of the root bark from Lycium chinense Miller, and its structure was determined using 1D and 2D NMR spectroscopy including DEPT, HMQC, and HMBC. (+)-Lyoniresinol-3alpha-O-beta-D-glucopyranoside exhibited potent antimicrobial activity against antibiotic-resistant bacterial strains, methicillin-resistant Staphylococcus aureus (MRSA) isolated from patients, and human pathogenic fungi without having any hemolytic effect on human erythrocytes. In particular, compound 1 induced the accumulation of intracellular trehalose on C. albicans as stress response to the drug, and disrupted the dimorphic transition that forms pseudo-hyphae caused by the pathogenesis. This indicates that (+)-lyoniresinol 3alpha-O-beta-D-glucopyranoside has excellent potential as a lead compound for the development of antibiotic agents. PMID- 16212234 TI - Anticoagulant 1,2,3,4,6-pentagalloyl-beta-D-glucopyranose isolated from geranium (Pelargonium inquinans Ait). AB - Geranium (Pelargonium inquinans Ait) leaves were extracted with 80% MeOH, and partitioned into n-hexane, ethyl acetate, BuOH and H2O to isolate the anticoagulant principles. The EtOAc fraction was found to be the most active, and was further purified using silica and octadecylsilane column chromatography employing a bioassay-guided fractionation method. The active compound was isolated and identified as 1,2,3,4,6-pentagalloyl-beta-D-glucopyranose (PGG) (compound I). The isolated anticoagulant significantly prolonged the activated partial thrombin time (APTT) and thrombin time (TT) using normal human plasma. One microgram of 1,2,3,4,6-pentagalloyl-beta-D-glucopyranose showed 0.063 heparin units in the APTT and 2.73 heparin units in the TT for anti-thrombosis. This is the first report of the isolation of PGG from geranium plants. PMID- 16212235 TI - Trichodermamide A and aspergillazine A, two cytotoxic modified dipeptides from a marine-derived fungus Spicaria elegans. AB - Two known modified dipeptides, trichodermamide A (1) and aspergillazine A (2), were isolated from an ethyl acetate extract of the metabolite of a marine-derived fungus Spicaria elegans, and were found to have a weak cytotoxic effect on three cancer cell lines P388, A-549, and HL-60 agreed. To our knowledge, this is the first report on the isolation of compounds 1 and 2 from the fungus Spicaria elegans and their cytotoxic effect. PMID- 16212236 TI - The cytotoxicity of eutigosides from Eurya emarginata against HL-60 promyelocytic leukemia cells. AB - Two phenolic glucosides, eutigoside B and eutigoside C were isolated from the fresh leaves of Eurya emarginata. These two phenolic glucosides exerted a significant inhibitory effect on the growth of HL-60 promyelocytic leukemia cells. Furthermore, when the HL-60 cells were treated with eutigoside C, several apoptotic characteristics such as DNA fragmentation, morphologic changes, and increase of the population of sub-G1 hypodiploid cells were observed. In order to understand the mechanism of apoptosis induction by eutigoside C, we examined the changes of Bcl-2 and Bax expression levels. The eutigoside C reduced Bcl-2 protein and mRNA levels, but slightly increased Bax protein and mRNA levels in a time-dependent manner. When we examined the activation of caspase-3, an effector of apoptosis, the eutigoside C increased the expression of active form (19-kDa) of caspase-3 and the increase of their activities was demonstrated by the cleavage of poly (ADP-ribose) polymerase, a substrate of caspase-3, to 85-kDa. The results suggest that the inhibitory effect of eutigoside C from E. emarginata on the growth of HL-60 appears to arise from the induction of apoptosis via the down-regulation of Bcl-2 and the activation of caspase. PMID- 16212237 TI - Cytotoxic saponins from the root of Dipsacus asper Wall. AB - Cytotoxic activitiy of seven hederagenin saponins isolated from the root of Dipsacus asper were investigated in vitro against L1210, HL-60 and SK-OV-3 tumor cell lines by the MTT method. 3-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L arabinopyranosyl hederagenin (2), 3-O-beta-D-xylopyranosyl-(1-->3)-alpha-L Rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl hederagenin (6) and 3-O-beta-D glucopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl hederagenin (7) exhibited the potent cytotoxicity against the three tumor cell lines with IC50 values ranging from 4.7 to 8.7 microg/mL, with the exception of compound 7, which exhibited weak cytotoxic activity against SK-OV-3 (IC50 22.5 microg/mL). Other compounds did not exhibit any cytotoxic activity (IC50 > 30 microg/mL). PMID- 16212238 TI - Pathophysiological implication of ganglioside GM3 in early mouse embryonic development through apoptosis. AB - Apoptosis may occur in early embryos where the execution of essential developmental events has failed, and gangliosides, sialic acid-conjugated glycosphingolipids, are proposed to regulate cell differentiation and growth. To evaluate the regulatory roles of ganglioside GM3 in early embryonic development, this study examined its expressional patterns in apoptotic cells during early embryonic development in mice. Pre-implanted embryos were obtained by in vitro fertilization, which were treated at the 4-cell stage with three the apoptosis inducers, actinomycin D, camptothecin and cycloheximide, for 15 h. All three inducers significantly increased the percentage of apoptotic cells, as measured using a TUNEL method, but remarkably reduced the total cell numbers. The numbers of morula and blastocyst stages were significantly decreased by treatment of the embryos with the three apoptosis inducers compared with the control, with a similar result also observed in the number of blastomeres. Staining of early embryos with Hoechst 33342 revealed a significant percentage of apoptotic nuclei. Prominent immunofluorescence microscopy revealed a significant difference in the ganglioside GM3 expression in apoptotic embryos compared with the control, and RT PCR also demonstrated a dramatic increase in ganglioside GM3 synthase mRNA in the apoptotic embryos. These results suggest that ganglioside GM3 may be pathophysiologically implicated in the regulation of early embryonic development through an apoptotic mechanism. PMID- 16212239 TI - Reductive depolymerization of bovine thyroglobulin multimers via enzymatic reduction of protein disulfide and glutathionylated mixed disulfide linkages. AB - The nascent thyroglobulin (Tg) multimer molecule, which is generated during the initial fate of Tg in ER, undergoes the rapid reductive depolymerization. In an attempt to determine the depolymerization process, various types of Tg multimers, which were generated from deoxycholate-treated/reduced Tg, partially unfolded Tg or partially unfolded/reduced Tg, were subjected to various GSH (reduced glutathione) reducing systems using protein disulfide isomerase (PDI), glutathione reductase (GR), glutaredoxin or thioredoxin reductase. The Tg multimers generated from deoxycholate-treated/reduced Tg were depolymerized readily by the PDI/GSH system, which is consistent with the reductase activity of PDI. The PDI/GSH-induced depolymerization of the Tg multimers, which were generated from either partially unfolded Tg or partially unfolded/reduced Tg, required the simultaneous inclusion of glutathione reductase, which is capable of reducing glutathionylated mixed disulfide (PSSG). This suggests that PSSG was generated during the Tg multimerization stage or its depolymerization stage. In particular, the thioredoxin/thioredoxin reductase system or glutaredoxin system was also effective in depolymerizing the Tg multimers generated from the unfolded Tg. Overall, under the net GSH condition, the depolymerization of Tg multimers might be mediated by PDI, which is assisted by other reductive enzymes, and the mechanism for depolymerizing the Tg multimers differs according to the type of Tg multimer containing different degrees and types of disulfide linkages. PMID- 16212240 TI - Mutant presenilin 2 increases acetylcholinesterase activity in neuronal cells. AB - A presenilin 2 mutation is believed to be involved in the development of Alzheimer's disease. In addition, transgenic mice with a presenilin 2 mutation have been reported to have learning and memory impairments. In this study, exposing PC12 cells expressing mutant presenilin 2 to 50 microM AP25-35, 30 mM L glutamate and 50 microM H2O2 caused a significant increase in acetylcholine esterase activity. An in vivo study revealed high levels of this enzyme activity in the mutant presenilin 2 transgenic brains compared with the wild type presenilin 2 transgenic and nontransgenic samples. These results suggest that a mutant presenilin 2-induced neurodegeneration in Alzheimer's disease might be involved in the increase in acetylcholinesterase activity. These findings might help in the development of an appropriate therapeutic intervention targeting mutant presenilin 2-induced Alzheimer's disease. PMID- 16212241 TI - Chitosan oligosaccharide inhibits 203HgCl2-induced genotoxicity in mice: micronuclei occurrence and chromosomal aberration. AB - The purpose of this study was to investigate the safety of chitosan oligosaccharide and the effects of chitosan oligosaccharide on mercury induced genotoxicity in mice using the micronuclei and chromosome aberration. The micronuclei test was performed by microscopic examination (x1,000, stained using a May-Grunwald solution) after administering 0.01, 0.1, and 1% (10 mg/mL) chitosan oligosaccharide for 7, 60, and 180 days ad libitum in mice. Total micronuclei of 1,000 polychromatic erythrocytes were recorded for each group. There was no difference between the untreated and experimental groups. The intake periods and concentrations of chitosan oligosaccharide did not affect the occurrence of micronuclei in bone marrow cells (P>0.05). The chromosomal aberration test was performed by microscopic examination (x1,000, stained using a 4% Giemsa solution) after administering the same concentration of chitosan oligosaccharide to mice, in F1, F2, F3 generations and parents. The frequency of chromosomal aberrations was defined as [Ydr = (D+R)/total number of counted lymphocytes]. Similar to the micronuclei test, there was no difference between the untreated and treated groups. These results showed that the intake periods and concentrations of chitosan oligosaccharide did not affect chromosomal aberrations in bone marrow cells (P>0.05). To investigate the effect of chitosan oligosaccharide on mercury-induced chromosome aberration, mice in each condition were supplied with 203HgCl2 and chitosan oligosaccharide ad libitum. Chitosan oligosaccharide significantly inhibited 203HgCl2-induced chromosome aberration in mice. Based on the results of this study, it may be concluded that the chitosan oligosaccharide is a nontoxic material that could be used as a suppressor of heavy metal-induced genotoxicity. PMID- 16212242 TI - Simultaneous determination of cannabidiol, cannabinol, and delta9 tetrahydrocannabinol in human hair by gas chromatography-mass spectrometry. AB - An analytical method was developed for evaluating the cannabidiol (CBD), cannabinol (CBN), and delta9-tetrahydrocannabinol (delta9-THC) level in human hair using gas chromatography-mass spectrometry (GC-MS). Hair samples (50 mg) were washed with isopropyl alcohol and cut into small fragments (< 1 mm). After adding a deuterated internal standard, the hair samples were incubated in 1.0 M NaOH for 10 min at 95 degrees C. The analytes from the resulting hydrolyzed samples were extracted using a mixture of n-hexane-ethyl acetate (75:25, v/v). The extracts were then evaporated, derivatized, and injected into the GC-MS. The recovery ranges of CBD, CBN, and delta9-THC at three concentration levels were 37.9-94.5% with good correlation coefficients (r2 >0.9989). The intra-day precision and accuracy ranged from -9.4% to 17.7%, and the inter-day precision and accuracy ranged from -15.5% to 14.5%, respectively. The limits of detection (LOD) for CBD, CBN, and delta9-THC were 0.005, 0.002, and 0.006 ng/mg, respectively. The applicability of this method of analyzing the hair samples from cannabis abusers was demonstrated. PMID- 16212243 TI - Preparation and in vivo evaluation of huperzine A-loaded PLGA microspheres. AB - Huperzine A-loaded microspheres composed of poly(D,L-lactide-co-glycolide) were prepared by an ONV emulsion solvent evaporation method. The characterization of the microspheres such as drug loading, size, shape and release profile was described. The in vitro release in the initial 7 days was nearly linear with 10% released per day. Thereafter drug release rate became slow gradually and about 90% drug released at day 21. The in vitro release rate determined by dialysis bag method had a good correlation with the in vivo release rate. Huperzine A aqueous solution was intramuscularly injected (i.m.) at 0.4 mg/kg and microspheres were intramuscularly injected at 8.4 mg eq huperzine A/kg in rats. The maxium plasma concentration (Cmax ) after i.m. microspheres was only 32% of that after i.m. solution. Drug in plasma could be detected until day 14 and about 5% of administered dose was residued at the injection site at day 14. The relative bioavailability of huperzine A microspheres over a period of 14 days was 94.7%. Inhibition of acyecholinesterase activity (AchE) in rat's cortex, hippocampus and striatum could sustain for about 14 days. In conclusion, huperzine A-loaded microspheres possessed a prolonged and complete drug release with significant inhibition of AchE for 2 weeks in rats. PMID- 16212244 TI - Formulation of microemulsion systems for transdermal delivery of aceclofenac. AB - An ONW microemulsion system was developed to enhance the skin permeability of aceclofenac. Of the oils studied, Labrafil M 1944 CS was chosen as the oil phase of the microemulson, as it showed a good solubilizing capacity. Pseudo-ternary phase diagrams were constructed to obtain the concentration range of oil, surfactant, Cremophor ELP, and co-surfactant, ethanol, for micoemulsion formation. Eight different formulations with various values of oil of 6-30%, water of 0-80%, and the mixture of surfactant and co-surfactant (at the ratio of 2) of 14-70%. The in vitro transdermal permeability of aceclofenac from the microemulsions was evaluated using Franz diffusion cells mounted with rat skin. The level of aceclofenac permeated was analyzed by HPLC and the droplet size of the microemulsions was characterized using a Zetasizer Nano-ZS. Terpenes were added to the microemulsions at a level of 5%, and their effects on the skin permeation of aceclofenac were investigated. The mean diameters of the microemulsions ranged between approximately 10-100 nm, and the skin permeability of the aceclofenac incorporated into the microemulsion systems was 5-fold higher than that of the ethanol vehicle. Of the various terpenes added, limonene had the best enhancing ability. These results indicate that the microemulsion system studied is a promising tool for the percutaneous delivery of aceclofenac. PMID- 16212245 TI - Study of the excess noise associated with demodulation of ultra-short infrared pulses. AB - The demodulation of ultra-short light pulses with photodetectors is accompanied by excess phase noise at the pulse repetition rate and harmonics in the spectrum of the photocurrent. The major contribution to this noise is power fluctuations of the detected pulse train that, if not compensated for, can seriously limit the stability of frequency transfer from optical to microwave domain. By making use of an infrared femtosecond laser, we measured the spectral density of the excess phase noise, as well as power-to-phase conversion for different types of InGaAs photodetectors. Noise measurements were performed with a novel type of dual channel readout system using a fiber coupled beam splitter. Strong suppression of the excess phase noise was observed in both channels of the measurement system when the average power of the femtosecond pulse train was stabilized. The results of this study are important for the development of low-noise microwave sources derived from optical "clocks" and optical frequency synthesis. PMID- 16212246 TI - Does the light shift drive frequency aging in the rubidium atomic clock? AB - Frequency aging in the rubidium (Rb) vapor-cell atomic clock plays a significant role in the device's timekeeping ability. Though many researchers have speculated on the physical mechanism(s) driving the linear, deterministic frequency change (i.e., deltaf(t)/fo = At), there is little unambiguous experimental data regarding the phenomenon. Here, long-term data were used from on-orbit global positioning system (GPS) Rb clocks to examine one postulated mechanism for frequency aging (i.e., the light-shift effect). Defining the light shift of the clock's fractional frequency as alphaI/Io, where alpha is the light-shift coefficient, we find that temporal variations of the relative light intensity, I/Io, cannot account for frequency aging. However, for the population of clocks considered here, we obtain the intriguing result that alpha/A = 1.7 +/- 1.5. Thus, it may be that frequency aging is driven by the light-shift effect through temporal variations of the light-shift coefficient. PMID- 16212247 TI - Effect of electromechanical coupling on static deformations and natural frequencies. AB - A two-way coupled electromechanical theory is used to study static deformations and free vibrations of a laminated hybrid rectangular plate comprised of either piezoceramic (PZT) layers or patches embedded at arbitrary locations in graphite/epoxy layers. A first-order shear deformation theory is used to develop equations for the plate which are solved by the finite-element method (FEM) using eight-node isoparametric elements. Static deflections and natural frequencies computed with open-circuited PZT layers are found to differ significantly from those of grounded PZT layers. PMID- 16212248 TI - Microwave de-embedding techniques applied to acoustics. AB - This paper describes the use of the microwave techniques of time domain reflectometry (TDR) and de-embedding in an acoustical application. Two methods of calibrating the reflectometer are presented to evaluate the consistency of the method. Measured and modeled S-parameters of woodwind instruments are presented. The raw measured data is de-embedded to obtain an accurate measurement. The acoustic TDR setup is described. PMID- 16212249 TI - Acoustic droplet vaporization for temporal and spatial control of tissue occlusion: a kidney study. AB - Acoustic droplet vaporization (ADV) has been introduced with the potential application of tumor treatment via occlusion and subsequent necrosis. New Zealand White rabbits were anesthetized, and their left kidney was externalized. An imaging array and single-element transducer were positioned in a tank with direct access to the kidney's vasculature and renal artery. Filtered droplet emulsions (diameter <6 microm) were injected intra-arterially (IA) into the left heart during insonification of the renal artery, and the extent of blood flow reduction by ADV was compared to the untreated right kidney. Flow cytometry (using colored microspheres) of kidney tissue samples and reference blood from the femoral artery allowed the quantitative estimation of regional blood flow. A maximum regional blood flow reduction in the treated region of >90% and an average organ perfusion reduction of >70% was achieved using ADV. After treatment of the left kidney, the control kidney on the contralateral side showed a maximum decrease in regional blood flow of 18% relative to the pre-ADV baseline. Image-based hyper echogenicity from ADV of IA injections was monitored for approximately 90 minutes, and cortex perfusion was reduced by >60% of its original value for more than 1 hour. This could be enough time for the onset of cell death and possible tumor treatment via ischemic necrosis. Moreover, currently used radiofrequency tissue ablation-based tumor treatment could benefit from ADV due to the decreased heat loss via vascular cooling. PMID- 16212250 TI - Evaluation of ultrasound tissue damage based on changes in image echogenicity in canine kidney. AB - Sufficiently high intensity ultrasound can create hyperechoic regions in an ultrasound image due to local bubble generation. We explore the link between the temporal extent of these hyperechoic regions and tissue damage caused by ultrasound therapy. The decay rate of increased echogenicity from the focal zone in insonated live exteriorized canine kidney was quantified and correlated to the spatial extent of tissue damage. The decay half-time, t(half), defined as the time for echogenicity enhancement to decay by a factor of 2, was observed in all cases to be greater than 41 s in spatial zones in which extensive histological damage was observed. In cases in which the measured thalf was less than 11 s, the damage was limited to minor hemorrhage, or it was not detected. These t(half) discrimination boundaries of 41 and 11 s were not statistically different for cases in which contrast agent was used to enhance therapeutic efficiency. This was true even though contrast agent infusion significantly reduced the therapy pulse duration threshold for damage production. PMID- 16212251 TI - Effects of contrast agent infusion rates on thresholds for tissue damage produced by single exposures of high-intensity ultrasound. AB - Stabilized microbubble ultrasound contrast agents (UCA) have potential to aid tissue ablation during ultrasound surgery by enhancing both cavitational and thermal damage mechanisms. Previously, we showed UCA infused at a rate of 1 microL/kg/min prior to ultrasound exposure could reduce the total energy required to produce tissue damage by up to two orders of magnitude. In this paper, we evaluate thresholds for macroscopic tissue damage with UCA infusion rates (IR) of 0.1, 0.3, 1, 3, and 10 microL/kg/min to determine IR potentially effective for ultrasound therapy. Canine kidneys were surgically externalized and insonified with single exposures of focused ultrasound. Incident exposures were 1.44 MHz tone bursts, either 250 ms in duration with intensity between 500 W/cm2 and 3200 W/cm2, or 100 micros to 1 s in duration with intensity equal to 3200 W/cm2. Probabilities of tissue damage occurrence were determined for each set of exposure conditions (intensity, duration, and IR). A threshold intensity and threshold duration, defined as the quantities for which tissue damage occurred with probability equal to 0.5, were estimated for each IR. Results show that, as IR increased from 0.1 to 10 microL/kg/min, the threshold intensity decreased by up to a factor of 3, and threshold duration decreased by up to a factor of 200. Microbubble introduction at IR up to 10 microL/kg/min thus may be effective in aiding ultrasound therapy. PMID- 16212252 TI - Adaptive imaging and spatial compounding in the presence of aberration. AB - Spatial compounding reduces speckle and increases image contrast by incoherently averaging images acquired at different viewing angles. Adaptive imaging improves contrast and resolution by compensating for tissue-induced phase errors. Aberrator strength and spatial frequency content markedly impact the desirable operating characteristics and performance of these methods for improving image quality. Adaptive imaging, receive-spatial compounding, and a combination of these two methods are presented in contrast and resolution tasks under various aberration characteristics. All three imaging methods yield increases in the contrast-to-noise ratio (CNR) of anechoic cysts; however, the improvements vary depending on the properties of the aberrating layer. Phase correction restores image spatial frequencies, and the addition of compounding opposes the restoration of image spatial frequencies. Lesion signal-to-noise ratio (SNR), an image quality metric for predicting lesion detectability, shows that combining spatial compounding with phase correction yields the maximum detectability when the aberrator strength or spatial frequency content is high. Examples of these modes are presented in thyroid tissue. PMID- 16212253 TI - Designed strength identification of concrete by ultrasonic signal processing based on artificial intelligence techniques. AB - This paper presents a pattern recognition method to identify the designed strength of concrete by evidence accumulation based on artificial intelligence techniques with multiple feature parameters. Concrete specimens in this experiment, which were designed to have the strengths of 180, 210, 240, 300, and 400 kg/cm2, respectively, have been considered. Variance, zero-crossing, mean frequency, autoregressive (AR) model coefficients, and linear cepstrum coefficients are extracted as feature parameters from ultrasonic signals of concretes. Pattern recognition is carried out through the evidence accumulation procedure using the distances measured with reference parameters. A fuzzy mapping function is introduced to transform the distance for the application of the evidence accumulation method. Results are presented to support the feasibility of the suggested approach for concrete pattern identification. PMID- 16212254 TI - Evaluation of glass materials by using the line-focus-beam ultrasonic-material characterization system. AB - We developed experimental procedures to evaluate glass materials using the line focus-beam ultrasonic-material-characterization (LFB-UMC) system. We prepared 28 specimens of a commercial borosilicate glass from random lots, and measured the velocities of leaky-surface acoustic waves (LSAWs) and leaky-surface-skimming compressional waves (LSSCWs), VLSAW and VLSSCW, using V(z) curve measurements at 225 MHz and 23 degrees C. The velocities for VLSAW ranged from 3121.83 m/s to 3149.77 m/s, with a maximum deviation of 27.94 m/s. The velocities for VLSSCW ranged from 5547.7 m/s to 5585.0 m/s, with a maximum deviation of 37.3 m/s. To investigate these observed variations in VLSAW and VLSSCW, we measured the bulk acoustic wave (BAW) properties, viz., longitudinal and shear velocities, then the densities and the chemical compositions of 8 of the 28 specimens. The LFB-UMC measurements confirmed that decreases in VLSAW and VLSSCW occur mainly with the B2O3 dopant concentrations, corresponding to the decrease of shear-wave and longitudinal-wave velocities that are caused by the decrease of the stiffness constants c44 and c11, respectively, rather than with decreased densities. The sensitivities are -6.36 x 10(-2) wt%/(m/s) for VLSAW and -4.87 x 10(-2) wt%/(m/s) for VLSSCW. This demonstrates that the LFB-UMC system is effective for evaluating glass materials and controlling production processes, by analyzing variations in chemical composition through the super-accurate velocity measurements of LSAWs and LSSCWs. PMID- 16212255 TI - Analysis of SAW properties in ZnO/AlxGa1-xN/c-Al2O3 structures. AB - Piezoelectric thin films on high acoustic velocity nonpiezoelectric substrates, such as ZnO, AlN, or GaN deposited on diamond or sapphire substrates, are attractive for high frequency and low-loss surface acoustic wave devices. In this work, ZnO films are deposited on AlxGa1-xN/c-Al2O3 (0 < or = chi < or = 1) substrates using the radio frequency (RF) sputtering technique. In comparison with a single AlxGa1-xN layer deposited on c-Al2O3 with the same total film thickness, a ZnO/AlxGa1-xN/c-Al2O3 multilayer structure provides several advantages, including higher order wave modes with higher velocity and larger electromechanical coupling coefficient (K2). The surface acoustic wave (SAW) velocities and coupling coefficients of the ZnO/AlxGa1-xN/c-Al2O3 structure are tailored as a function of the Al mole percentage in AlxGa1-xN films, and as a function of the ZnO (h1) to AlxGa1-xN (h2) thickness ratio. It is found that a wide thickness-frequency product (hf) region in which coupling is close to its maximum value, K(2)max, can be obtained. The K(2)max of the second order wave mode (h1 = h2) is estimated to be 4.3% for ZnO/GaN/c-Al2O3, and 3.8% for ZnO/AlN/c-Al2O3. The bandwidth of second and third order wave modes, in which the coupling coefficient is within +/- 0.3% of K(2)max, is calculated to be 820 hf for ZnO/GaN/c-Al2O3, and 3620 hf for ZnO/AlN/c-Al2O3. Thus, the hf region in which the coupling coefficient is close to the maximum value broadens with increasing Al content, while K(2)max decreases slightly. When the thickness ratio of AlN to ZnO increases, the K(2)max and hf bandwidth of the second and third higher wave modes increases. The SAW test devices are fabricated and tested. The theoretical and experimental results of velocity dispersion in the ZnO/AlxGa1 xN/c-Al2O3 structures are found to be well matched. PMID- 16212257 TI - A flexible piezoelectric transducer design for efficient generation and reception of ultrasonic Lamb waves. AB - This paper describes the development of a flexible piezoelectric transducer for the generation and detection of ultrasonic symmetrical Lamb waves in plate-like structures. This piezoplatelet transducer structure comprises an array of miniature piezoceramic plates embedded within a soft setting polymer filler material, combining the efficiency of the active piezoceramic phase with a degree of flexibility, which is a function of the platelet/polymer dimensions. For many condition-monitoring applications, the generation of ultrasonic Lamb waves is often appropriate, and this was achieved by incorporating interdigital design techniques via the transducer electrode pattern. The performance of the piezoplatelet transducer structure was evaluated using a combination of linear systems and finite-element modeling, substantiated by experimental results. Importantly, the transducer is shown to operate as an ensemble of platelets, each operating in the thickness mode and well decoupled from neighboring piezoelectric elements. Using this transducer configuration, an unimodal s1 Lamb wave, at 1.45 MHz, has been generated and detected in a 3-mm thick steel plate. Furthermore, a propagation distance of almost 1 m was recorded for s0 Lamb wave generation/detection in a fiber-reinforced composite plate. PMID- 16212256 TI - Synthesis of C-axis-oriented AlN thin films on high-conducting layers: Al, Mo, Ti, TiN, and Ni. AB - Thin piezoelectric polycrystalline films such as AlN, ZnO, etc., are of great interest for the fabrication of thin film bulk/surface acoustic resonators (TFBARs or TFSARs). It is well-known that the degree of c-axis orientation of the thin films correlates directly with the electromechanical coupling. However, the degree of c-axis orientation of the piezoelectric film is, in turn, influenced by other parameters such as the structure of the substrate material, the matter of whether the c-axis is up or down (polarity), and the growth parameters used. The correlation of these three aspects with the electromechanical coupling of the AlN thin films, is studied here. Thin AlN films, prepared in a magnetron sputtering system, have been deposited onto thin Al, Mo, Ni, Ti, and TiN films. Such thin high-conducting layers are used to form the bottom electrode of TFBAR devices as well as to define a short-circuiting plane in TFSAR devices. In both cases, they serve as a substrate for the growth of the piezoelectric film. It has been found that the degree of orientation and the surface roughness of the bottom metal layer significantly affects the texture of the AlN films, and hence its electroacoustic properties. For this reason, the surface morphology and texture of the metal layers and their influence on the growth of AlN on them has been systematically studied. Finally, FBARs with both Al and Ti electrodes have been fabricated and evaluated electroacoustically. PMID- 16212258 TI - Different effects of microbubble destruction and translation in Doppler measurements. AB - In flow measurements in which microbubbles are involved, the amplitude and phase of the received echo signal are noticeably influenced by the transmitted ultrasound intensity. Previous studies have shown that, when such intensity is progressively increased, the Doppler spectrum is accordingly distorted, i.e., it is asymmetrically broadened toward the negative frequency side. Such deformation has been attributed to radiation force, which pushes the microbubbles into the sound propagation direction, thus yielding additional phase delays in the received echoes. However, the possible contribution of microbubble destruction to this spectral deformation has not been considered yet. In this paper, this issue is investigated by analyzing the experimental spectra produced by two different types of microbubbles suspended in a moving fluid and insonified in pulsed wave (PW) mode at programmable pulse repetition frequency (PRF) and pressure. Conditions are created in which either the radiation force or the destruction mechanism is expected to be dominant. Effects produced by the two phenomena on the Doppler spectrum are shown to be different. When the PRF is low (2 kHz), so that, according to theoretical simulations, the radiation force effect is negligible, a 26 dB noise floor increase is observed for a 13 dB pressure increment. For a higher PRF (16 kHz), the same pressure increase not only affects the noise floor, but also causes the bubbles to deviate from their original streamlines, yielding a Doppler bandwidth increase by a factor of 5. It is concluded that asymmetrical spectral broadening is mainly due to radiation force, and microbubble destruction mainly results in an increased noise floor without affecting the spectral shape. PMID- 16212259 TI - Suppression of spurious lateral modes in thickness-excited FBAR resonators. AB - Thin film bulk acoustic wave resonators (FBAR) utilize thickness-excited modes in which the resonant frequency is determined by the thickness of the structure and the wave velocity of the mode used. Unfortunately, other resonant modes also may be excited in the device. Some of these correspond to low-frequency, laterally excited modes and, although a relatively small amount of the total energy is absorbed by these modes, their harmonics may produce an undesirable response around the fundamental resonance frequency of the desired thickness mode. This work explores various ways of suppressing the spurious effects caused by lateral excited modes by studying their dependence of the electrode geometry. The origin of the lateral-excited modes is discussed in detail, and the results from a number of different electrode geometries are compared. A new elliptical electrode shape for suppression of spurious modes is developed and demonstrated. PMID- 16212260 TI - [Homologous protein domains in superkingdoms Archaea, Bacteria, and Eukaryota and the problem of the origin of eukaryotes]. AB - The distribution of protein domains was analyzed in superkingdoms Archaea, Bacteria, and Eukaryota. About a half of eukaryotic domains have prokaryotic origin. Many domains related to information processing in the nucleocytoplasm were inherited from archaea. Sets of domains associated with metabolism and regulatory and signaling systems were inherited from bacteria. Many signaling and regulatory domains common for bacteria and eukaryotes were responsible for the cellular interaction of bacteria with other components of the microbial community but were involved in coordination of the activity of eukaryotic organelles and cells in multicellular organisms. Many eukaryotic domains of bacterial origin could not originate from ancestral mitochondria and plastids but rather were adopted from other bacteria. An archaeon with the induced incorporation of alien genetic material could be the ancestor of the eukaryotic nucleocytoplasm. PMID- 16212262 TI - [Activity of lectins of winter wheat seedlings infected with mycoplasma and treated with salicylic acid]. AB - Changes in activity of soluble and cell wall lectins have been revealed in seedlings of winter wheat Triticum aestivum L. cultivar Mironovskaya 808 after infection with mycoplasma Acholeplasma laidlawii 118. The protective effect of salicylic acid was manifested as negating the bursts of lectin activity induced by mycoplasma infection. PMID- 16212261 TI - [Activation of deoxyribonucleotide synthesis by radioprotectants and antioxidants as a key stage in formation of body resistance to DNA-damaging factors]. AB - The responses of the systems of synthesis of deoxyribonucleotides (dNTPs), DNA, and proteins in hematopoietic organs and liver of animals to gamma-radiation, administration of radioprotectants and antioxidants as well as the dependence of these responses on the doses of radiation and drugs were studied. Radioprotectants of acute (indralin) and durable effects (indomethaphen) as well as natural (alpha2-tocopherol) and synthetic anti-oxidants (ionol or 2,6-di-tert butyl-4-methylphenol) efficient in survival test were used. Three stages could be recognized in the standard unspecific response of the studied systems to radiation: (1) immediate increase in ribonucleotide reductase activity in the tissues within the first 30 min as a part of the integrated SOS response to DNA damage, which activates dNTP synthesis; (2) inhibition of the synthesis of dNTPs, DNA, and and (3) restoring ribonucleotide reductase activity and integral increase in the production of dNTPs, DNA, and total protein, which is essential for the development of compensatory and restorative responses of the organism. The radioprotectants significantly increased ribonucleotide reductase activity, which increased intracellular concentrations of the four dNTP types in organs during radiation exposure and three following days. Within this period, ribonucleotide reductase activity was inhibited by 40-50% in animals not treated with radioprotectants as compared to control. Balanced high pools of dNTPs in the organs of radioprotectant-treated animals provided for high-performance repair of DNA damage. The radioprotectant-induced activation of dNTP synthesis during the development of compensatory and restorative responses provides for an earlier restoration of the cellular composition and functioning of the organs. Antioxidants stimulated the synthesis of dNTPs, DNA, and proteins in animal tissues in a strict dose interval. Their effect on the studied syntheses was dose dependent: single or multiple long-term administration of high antioxidant doses inhibited synthesis of dNTPs, DNA, and proteins. Radioprotectants and antioxidants affected the pool of blood protein Fe3+-transferrin controlling the synthesis of iron-containing ribonucleotide reductase activity in hematopoietic organs, and hence, the iron-dependent stage in DNA synthesis--dNTP synthesis. Activation of protein synthesis in organs by the studied substances increased the pools of Fe3+-transferrin and Cu2+-ceruloplasmin in the blood, which activated dNTP and DNA synthesis. Activated synthesis of dNTP, DNA, and proteins in the organs and increased pools of studied plasma proteins underlay the formation of body resistance to DNA-damaging factors. PMID- 16212263 TI - [Comparative morphological properties of different hepatoid glands: histological variants and the morphofunctional unit]. AB - Three groups can be recognized in the type of hepatoid glands: glands synthesizing protein alone, glands synthesizing protein and hydrophobic lipids and/or melanin, and intermediate glands. Histological variants (seven in total) can be recognized within each group. Different possible combinations of hepatoid glands with other derivatives of hair follicles are considered. The first definition of the morphofunctional unit of the hepatoid land is proposed. PMID- 16212264 TI - [A paradox of a parasite prolonging the life of its host. Pearl mussel can cancel the accelerated aging program in salmon]. AB - A unique case is analyzed when the accelerated aging program (progeria) in salmons (Salmonidae) can be canceled by larval parasite of the gills--freshwater pearl mussel Margaritifera margaritifera. As a result, the maximum age of Salmo fishes hosting the mussel can be as high as 13 years. The mollusk-fish system made it possible to demonstrate that the parasite can inhibit aging of the host and stimulate nonspecific resistance to stress, i.e., can control longevity. The mussel proved to increase the resistance to epitheliomata and cutaneous mycoses. The parasite is perceived to neutralize the senile changes in the regulatory system hypothalamus-pituitary-peripheral endocrine glands-hypothalamus of salmon. PMID- 16212265 TI - [Intraspecific genetic and morphological variation of earthworm Eisenia foetida (Sav.)]. AB - Polyacrylamide gel electrophoresis was used to study the allelic frequency distribution of four polymorphic loci (Pox, Lap, Pgm, and Odh) in 22 samples of Eisenia foetida (Sav.) from distant parts of the range: European Russia, Ukraine, Kazakhstan, and Kyrgyzstan. A hierarchical population structure was demonstrated for this species: local populations are integrated into spatial groups. Statistical analysis of morphological variation demonstrated that recognition of the spatial groups on the basis of biochemical and genetic characters as individual taxa of any rank is not justified. PMID- 16212266 TI - [Development of the method of magnetic neutron capture therapy of cancer]. AB - The method of magnetic neutron capture therapy (MNTC) of cancer can be described as a combination of two methods: the targeted delivery of drugs using magnetic carriers and the proper neutron capture therapy which consists in tumor irradiation with thermal neutrons following the delivery of 10B compounds to the tumor site. Two-component ultradispersed particles containing Fe and C were tested as magnetic adsorbents of boron phenylalanine and borax. The quantities of absorbed borax proved sufficient for high concentration of boron atoms at the tumor site. The kinetics of boron release to saline substantiates the application of Fe-B (10%) ultradispersed particles for efficient MNTC. Both particle types have high magnetization and magnetic homogeneity, can form stable magnetic suspensions, and have low toxicity. PMID- 16212267 TI - [Contribution of the endogenous opioid system to regulation of heart resistance to the arrhythmogenic effect of short-term ischemia and reperfusio]. AB - Preliminary selective blockade of micro, delta1, delta2, kappa1, and kappa2 opioid receptors proved to have no effect on the incidence of ventricular arrhythmias during a 10-min coronary occlusion and subsequent reperfusion in ketamine-anesthetized rats. We propose that the endogenous opioid system has no considerable role in regulation of heart resistance to the arrhythmogenic effect of short-term local ischemia and subsequent reperfusion. PMID- 16212268 TI - [Effect of modification of the N-terminal region of molecule on the expression of neotropic effect of semax analogues]. AB - A comparative study of neotropic activity of semax (MEHFPGP), an analogue of the ACTH(4-10), and some of its derivatives in which the N-terminal methionine was modified or substituted with other amino acid residues was performed. The effect of these peptides on learning of albino rats in tests with positive (alimentary) and negative (pain) reinforcement was studied. In the case of modification of methionine by attachment of the gluconic-acid residue or substitution of methionine with lysine, the neotropic effect of the peptide was retained. The substitution of methionine with tryptophan or serine resulted in a decrease in the neotropic activity. The substitution of methionine with glycine, threonine, or alanine caused a complete loss of the neotropic activity of the peptide. Therefore, the amino acid residue located at position 1 of the heptapeptide analogue semax, plays a key role in retaining the neotropic effects of the peptide and determines the degree of their expression. PMID- 16212269 TI - [Variation of ecological status of Norwegian Sea water determined from hydrolytic enzyme activities]. AB - The capacity of biopolymer transformation involving efficient and highly specific natural enzyme mechanisms was studied in seawater of the dynamic zone of the Norwegian Sea (the Voring Plateau region). Vertical and spatial variation of proteinase and amylase activities was demonstrated in seawater and the potential rates of degradation of specific substrates, azocasein and Procion-5CX-modified starch, were calculated. High proteolytic activity was demonstrated for the upper photic layer (0-10 m) in the southwestern part of the polygon (up to 88 U/l; v(pr) = 7.04 mg/l/h). Proteolytic activity in the abyssal layer (1500 m and below) sharply decreased to 8-16 U/1; v(pr) = 0.64-1.28 mg/l/h. Similar to other regions of the ocean, the pattern of amylase activity in seawater included low rates of polysaccharide destruction (0-4 U/I; v(st) = 0-0.2 mg/l/h) in water with high proteolytic capacity and, conversely, the top amylase activity (up to 246 490 U/l; v(st) 12.3-24.5 mg/l/h) in seawater layers with undetectable or low proteolytic activity. The spatial distribution of the enzyme activities can indicate the presence of water bodies of different genesis. In the southwestern part of the polygon, statistical analysis demonstrated high correlation between hydrophysical indices (temperature, salinity, and salinity gradient) and proteinase and amylase activities. The ecological evaluation based on express enzyme-substrate tests demonstrated a stressful situation for destruction of organic compounds of protein nature in both the photic layer and the layers below 1000 m (t(pr) > or = 10 h). PMID- 16212270 TI - [Functional significance and weight properties of the shell in certain mollusks]. AB - Functional, morphological, and weight properties of the shell are discussed for many gastropods and bivalves living under various conditions. PMID- 16212271 TI - [Application of fungal metabolites for evaluation of pine stand vigor in conditions of industrial pollution]. AB - Two methods for evaluation of pine stand vigor were tested on permanent sample plots near Krasnoyarsk City: visual evaluation and exposure to fungal metabolites. In the first case, forest vigor was estimated using the six-point system of the Sanitary Regulations of the Russian Federation as the mean score for 200 trees on the sample plots. In the second case, the vigor was evaluated from the size of the necrotic spot on the tree cortex after inoculation of 22-25 randomly chosen pines with the extract of fungus Ceratocystis laricicola Red-fern and Miter. The necrotic spot size was significantly larger in pines from polluted forest as compared to background one. This points to the decrease in infection protection of trees affected by pollution, although visual evaluation of the stand vigor could not distinguish the polluted and background stands. PMID- 16212272 TI - Morphometric comparison of human nerve cells: pyramidal motor system. AB - We compared morphological and morphometric data on various motor neurons in the human pyramidal system using the modified Kluver-Barrera staining method with extremely minimized shrinkage ratio and an image-analyzer. We classified motor neurons in the human pyramidal system into three groups according to the measurement data. This report may be of interest to better understand the process of nerve conduction in the human pyramidal system. PMID- 16212273 TI - Morphometric evaluation of posterior funiculus nerve fibers in relation to aging. AB - We morphologically evaluated the size of axons in the posterior funiculus in different age groups and examined the changes due to aging. In the past, such studies have been conducted at the cervical spinal cord (C6) level, and a decrease in the size and number of axons due to aging has been noted. The current study was conducted at the lower lumbar spinal cord (L2) level. PMID- 16212274 TI - Morphological characteristics of nerve cells in the human general sensory system. AB - We conducted a study to compare morphological characteristics of various nerve cells in the sensory system. This sort of evaluation is indispensable for a better understanding of the nervous system in relation to clinical physiology and neurology. PMID- 16212275 TI - Morphometric comparison of human spinal cord anterior horn cells between the cervical and lumbar levels. AB - Morphometric comparisons of anterior horn cells (AHCs) were conducted between cervical and lumbar spinal cords in 29 cadavers. The sizes of the AHCs were always larger at the lumbar level than at cervical level irrespective of the age of the spicemens, but a decrease in size with age was observed at both levels. The correlation coefficient shows a larger decrease in numbers and volumes at lumbar level than at cervical level. PMID- 16212276 TI - NADH dehydrogenase activity and expression of mRNA of complex I (ND1, 51kDa, and 75kDa) in heart mitochondria of klotho mouse. AB - Mitochondrial enzyme activities and ultrastructure of mitochondria prepared from klotho mutant mice were compared with those in wild-type mice. We also measured the levels of expression of ND1, 51kDa, and 75kDa mRNA associated with the genes encoding NADH dehydrogenase and complex I and that of alpha cardiac myosin heavy chain mRNA in both groups. Mitochondrial NADH oxidoreductase activity was higher in klotho mutant mice during aging than that in wild-type mice. The area of mitochondria per unit area (300 microm2) of cell was almost constant from 4 to 7 weeks of age in both groups. A few large mitochondria were scattered between numerous small mitochondria with compact cristae and myofibrils in klotho mice from 5 weeks of age. The levels of ND1 and 75kDa mRNA were slightly high from 7 weeks of age in klotho mutant mice, whereas they were almost constant in wild type mice, in spite of reduced expression of alpha cardiac myosin heavy chain mRNA. Our results indicate that klotho protein indirectly plays a role in diminished functional adaptability of enzymes in aged heart muscle, and is required for hypertrophy of cardiac mitochondria. PMID- 16212277 TI - Induction of corneal lesion and nerve fiber sprouting by neonatal capsaicin application depends on the dose of the drug and survival time after treatment. AB - The effects of capsaicin on the rat cornea and its NsAchE (non specific acetylcholinesterase)-positive nerve fibers were investigated after long and short survival periods following subcutaneous (s.c) injection of the drug. Sixteen rats were injected once s.c with capsaicin on postnatal day 2 at a dose of 50 mg/kg. Age-matched rats were injected only vehicle, and served as control (n = 16). After 4 (n = 6), 8 (n = 6) and 12 (n = 4) mos., both sides of the corneas were examined under a binocular microscope to look for corneal abnormalities. Immediately after the enucleation, bilateral corneas were excised with a thin scleral margin and their ciliary body and iris were removed in DPBS solution. Then, they were fixed in a 4% paraformaldehyde solution containing 8% sucrose for 1 h at 4 degrees C, and processed for staining by the NsAchE method. For comparison, 15 rats were injected 3 times with capsaicin (total dose: 150 mg/kg) at a dose of 50 mg/kg on days 1, 2 and 3 after birth. Age-matched rats (n = 16) were injected vehicle as controls. Five (n = 4), 17 (n = 4), 45 (n = 3) and 75 (n = 4) days later, their corneas were similarly handled as described above. Corneal lesions and sprouting of the NsAchE-positive subepithelial nerve fibers appeared 4 mos. after the treatment with capsaicin (50 mg/kg). In particular, all the treated corneas (8/8) at 12 mos. showed corneal abnormalities. Contrary to the results from the single injection of capsaicin, corneal changes had already appeared on day 17 after treatment with capsaicin at a high dose (150 mg/kg). These data reinforce the suggestion that yielding of corneal lesions is closely associated with the longer survival time at lower doses and with the short survival time at a high dose of the drug. Further, the relationship between sprouting of corneal nerve fibers and corneal wounds was discussed on the basis of the previous reports. PMID- 16212278 TI - Morphological classification of mandibular dental arch forms by correlation and principal component analyses. AB - To evaluate the morphology of dental arches, 53 (male: 29, female: 24) paired casts having normal dentitions and occlusion were selected from 396 (age: 18 to 26 years old; male: 257, female: 139) sets of dental study models. The mandibular dentitions were preliminarily classified as square, round-square, round and round V-shaped arches based on the conventional morphological descriptions. Midpoints of the incisor edge (I1R, I1L, I2R, & I2L), summits of the cuspids (CR & CL), buccal cusps of the premolars (P1R, P1L, P2R, & P2L), mesial buccal cusps of the first and second molars (M1R, M1L, M2R, & M2L), and the midpoint (A) of line I1R I1L were designated as reference points. From A, let a vertical line intersected line M2R-M2L at reference point B. The line A-B intersected CR-CL at reference point E. We evaluated 1) the protrusion of the cuspids by 1. angle I2R-CR-P1R (angle R) + angle I2L-CL-P1L (angle L); 2) the curvature of the anterior teeth by 2. (A-B)/(CR-CL), 3. (180 degrees-angle(CR-A-CL), and 4. (A-E)/(CR-CL); 3) the length to width ratio of the dental arch by 5. (A-B)/(M2R-M2L); 4) the degree of roundness of the mandibular arch by estimation of 6. (rtheta5 - rtheta4)R + (rtheta5 - rtheta4)L; and 5) an item 7. for the differentiation of type I and type II round-square arches by relating the bilateral contour and position of break line P1-P2-M1-M2 (i) to line P1-M2 (ii). The data of items 1., 2., 3., 4., 5., and 6. were further standardized and summarized into three essential principal components: 1) the curvature of the anterior teeth, 2) the curvilinear contour of the dental arch, and 3) the length-to-width ratio of the dental arch. The results indicated that: 1) 36 cases (67.9%) of the mandibular dentitions were round-square arches which showed no prominent principal component. 11 cases (20.8%) were square arches and 6 cases (11.3%) were round V-shaped arches; no round arches was found in mandibular dentitions. 2) Statistical analysis indicated significant differences of items 3., 4. and 6. in various mandibular arches (Student's t-test). 3) By examination of the three principal components, significant differences of item 5. between the round V-shaped arches and square and round-square mandibular arches were evident (Student's t-test). The present study elucidated that morphology of the mandibular arch was determined by a parameters representing the curvature of anterior teeth (composed of items 2., 3. and 4., and another parameter (item 6.) representing roundness of the mandibular arch. PMID- 16212279 TI - [Animals (Animalia) in the system of organisms. 1. Typological systems]. AB - Since times of Aristotle animals were considered as a group, opposing to plants. The last were distinguished by two characters. Plants as distinct from animals live the attached way of a life and all nutrients receive from a substratum on which live and from the surrounding air. Animals live an active way of life and exist due to digestion. Fungi at such definition belong to plants. Only in second half of XX centuries due to works of Whittaker and of Tachtadjan fungi have received the separate status equally with plants and animals. In this new system of a plant embraced either oxygenic phototrophs, or photosynthetic eukaryotes. The traditional characters distinguishing animals from plants and fungi are in detail analysed. Many of them appeared formal, not reflecting the structure of relationship. Comparing heterotrophs some authors saw in absorptive nutrition the main difference of fungi from animals. However on mechanisms of receipt of substances in a cell fungi, animals and plants do not differ. Phagocytosis and pinocytosis (clathrin-mediated endocytosis), considered as the most characteristic feature of animals, are revealed both in fungi, and in plants. On photosynthetic activity plants form heterogeneous group, differing on primary and secondary plastids. The last besides have the various origin connected to symbiogenesis of the host cell with red or green algae. Heterotrophy cannot be considered as a uniting attribute of fungi and animals. It is essentially different and focused on diverse food sources. Evolution of animals is connected to perfection of structure of a plasmatic membrane and saturation by its molecules allowing a cell, and through it all organism to be guided in an environment and adequally to be up to external irritants. At a cellular level animals use the various mechanisms of cellular activity connected to moving of cells, their combination in aggregates and complexes or, on the contrary, separation in new cellular configurations. The complex of cellular adaptations connected to the analysis of external signals and adequate response to them of cells, underlies the phenomenon of irritability. At a cellular level irritability is mediated through work of the actin apparatus. Lamarck in "Philosophie zoologique" considered irritability as the main distinctive feature of animals. Evolution of plants and fungi went in a direction of development of a secondary metabolism. The secondary metabolism, concerning synthesis of protective substances, is peculiar to all sedentary organisms, including the animals. PMID- 16212280 TI - [Evolution as resistance to entropy. II. A conservative role of the sexual reproduction]. AB - It is claimed that biological meaning of the sex and meiotic genetic recombination is a creation of a barrier for evolution. The transition to sexuality is not merely a change in reproduction mode but a leap to a new known as cohesion. In a sexual population, the lineages of different individuals become tangled into multidimensional net, resulting in a creation of gene pool and new superindividual entity--biological species. A sexual individual can not reproduce its particular genocopy and its fitness is sacrificed to some extent for the fitness of the species. The competition between individuals is replaced with the competition between gene alleles given that the competitors have after all the common offspring. The genotype of the "outstanding" individuals with a highest fitness are not transmitted to next generation, being scattered and shuffled in new combination after unavoidable crossing with the "ordinary" partners. So, the sexual reproduction can evolve only as a whole. Genetic recombination in meiosis changes a character of mutations distribution among gametes enhancing the classes with mutation load both lower and higher than average. By this, an efficiency of the truncated selection (elimination of the individuals with multiple mutations) is enhanced and an ability to restore the initial genotype appears. Evolution within the species becomes reversible, which is equivalent to its virtual cessation. The species acquires an evolution resistance that can be overcome by rare concurrence of circumstances. PMID- 16212281 TI - [Genetic mechanisms of realization of the law of limiting factor in Saccharomyces cerevisiae]. AB - The law of limiting factor establishes the dependence organism functioning on the environmental conditions. The molecular basis of this adaptation determined with differentional gene's activity. The expression of more than 6000 genes in response to the deficiency in nitrogen, carbon or phosphorus in growth medium has been studied in yeast Saccharomyces cerevisiae. The model of integral yeast cell response on limiting factor was suggested on the basis of this data and the data concerning mechanisms of metabolic pathways regulation. According to this model, phosphoprotein kinase (Tor1p, Tor2p, Snf1p or Pho85p) that regulates corresponding metabolic pathway receives a signal about the limiting factor and become dominated kinase. Dominated kinase regulates activators and repressors proteins of its own metabolic pathway but it is also able to affect the activity of transcription regulators of other metabolic pathways. Proteins could be activators of one set of metabolic pathways and in the same time serve as repressors of others. That allows the regulation of cell metabolism according to the limiting factor. Dominated kinases can be involved in phosphorylation of DNA polymerases and affect the level of spontaneous mutations. Mutations that increase that viability of cells on the background of limiting factor are selected. Thus the relationship between inheritable and non-inheritable variability is not occasional and take place on the genetic level and the limiting factor is the reason for modification and inheritable variabilities. PMID- 16212282 TI - [The validity of the Willis Law for taxonomy]. AB - Different explanations of Willis' law are discussed. The common view on the observed relationship between species and genus frequencies suggests that Willis' rule is artifact of two independent processes. However, actual data on similarity of the diversity patterns in different taxa contradicts with this of view and show that there are intrinsic laws regulating biological diversity. The analysis of statistical parameters of diagnostic keys reveals that distribution of differential features according to the frequency of their usage appeas to be ranked. Considering taxon size as extensional characteristic and the set of differential features as intensional characteristic one can conclude that nature of disctribution is identical in both cases. Existed data indicates that biological diversity is self-organizated and self-ordered. It allow to reconsider the conception of Natural system which could be considered as fractal with the properties of the whole composed of parts (taxa) similar by their properties to each other and to the whole. The lowest parts, units of Natural system are taxa of generic rank, their characteristics is subjected to the Willis' rule. PMID- 16212283 TI - [Acoustic communication and sexual selection in Orthoptera (Insecta)]. AB - Comparison of calling and courtship songs and mating strategies in different groups of Orthoptera shows that acoustic signals in bush crickets (Tettigonioidea) are used for searching conspecific mates at a distance, and song evolution is primarily driven by the acoustic surroundings and simultaneously singing other species. The role of sexual selection in the evolution of acoustic signals in bush crickets is much less important than in the evolution of other nonacoustic signals employed during direct contact. Acoustic signals of crickets (Grylloidea) are involved in both distant and short-range communication. Acoustic courtship signals, along with other courtship components, may offer the material for sexual selection, although, essentially, evolution of acoustic signals in crickets is determined by the acoustic surroundings. Acoustic communication in the grasshoppers of the subfamily Gomphocerinae is mainly a short-distance communication. Acoustic signals of Gomphocerinae are highly variable and elaborate in temporal parameters, and therefore can serve for evaluation of the mate "quality". Song evolution in this group is to a greater extent driven by sexual selection than by the acoustic surroundings and, therefore, could proceed faster than in other groups of Orthoptera and play a major role in speciation. PMID- 16212284 TI - [Structure, occurrence and functional significance of nonlinear phenomena in sounds of terrestrial mammals]. AB - Subharmonics, deterministic chaos, biphonations, sidebands and frequency jumps, known under joining name nonlinear phenomena, represent acoustical appearances that occur in calls of various mammals, from insectivorous to humans. Although the physical basis for appearances of the nonlinear phenomen is known in principle, such aspects as occurrence and functional significance of nonlinear phenomena, are poorly understood. We described here the structural peculiarities of these appearances based on domestic dogs' whines demonstrating all kinds of the nonlinear phenomena. The nonlinear phenomena result directly from the work of mammalian vocal apparatus--lunges, vocal folds, and vocal tract, which are responsible for its inherent functional characteristics. The mammalian vocal folds represent a system of two coupled oscillators functioning in different vibratory regimes depending on degree of synchronization in their vibrations and occurrence of coupling between the vocal folds. This functioning does not need in direct neural control for turning on the complex regimes of vocal folds' vibration and switching from one regime to another. Besides, many mammals possess anatomical structures, such as vocal membranes, pads on vocal folds, or laryngeal air sacs, that are to participate in sound production as additional oscillators extend the range of possibilities for arising of nonlinear phenomena in vocalization. On the basis of published and own data, we provide and overview of the occurrence of nonlinear phenomena in sounds of humans, nonhuman primates, canids, and rodents, and discuss the supposed functional significance of these acoustical appearances in mammalian vocal communication systems. From one side, nonlinear phenomena may be related to various physiological disorders in humans and animals. In such cases, their appearance in calls may be nonadaptive, because they permit conspecifics' to avoid owners of such "ill" voices, and points easy prey to predators. From another side, in some cases the nonlinear phenomena may arise especially for performing of some signal functions in species communication system: to enhance reliability of individual recognition, to transmit information about size of a caller over the large distance, to permit individuals of not great size mimicry acoustically under more larger animal, to introduce variety into monotonous vocal sequences in order to force other group members to pay attention to a caller, and to facilitate distance estimation to a caller and direction of its movement. PMID- 16212285 TI - Optimized prolate ablations with the NIDEK CXII excimer laser. AB - PURPOSE: To describe a novel excimer laser ablation algorithm, termed optimized prolate ablation (OPA), that uses both topography and ocular aberrometry and maintains or improves the natural corneal shape postoperatively. METHODS: A descriptive article outlining the theory behind this OPA algorithm. RESULTS: The theoretical changes to the ablation algorithm described in this article will produce a prolate cornea postoperatively. CONCLUSIONS: OPA treated eyes will have equal or better visual quality than preoperatively because age-related changes from lenticular spherical aberration are measured and treated. PMID- 16212286 TI - Optimized prolate corneal ablation: case report of the first treated eye. AB - PURPOSE: To present the first patient treated with an optimized prolate ablation algorithm (OPA) with the NIDEK Advanced Vision Excimer laser platform (NAVEX; NIDEK, Gamagori, Japan). METHODS: A patient with moderate myopia and large scotopic pupils received contralateral LASIK treatment, treating one eye with OPA and the other with conventional treatment. RESULTS: At 3 months postoperatively, corneal topography and wavefront analysis show a prolate result with the OPA treatment and an oblate result with the conventional treatment. CONCLUSIONS: The creation of prolate cornea over the entire scotopic pupil, the reduction of spherical aberrations, and an ablation architecture that compensate for age related changes was achieved immediately after LASIK. The OPA treatment may result in better visual quality over the long-term compared to conventional LASIK. PMID- 16212287 TI - Objective quality of vision in presbyopic and non-presbyopic patients after pseudoaccommodative advanced surface ablation. AB - PURPOSE: To analyze the objective quality of vision at 6 months postoperatively after pseudoaccommodative (presbyopic) advanced surface ablation (PASA). METHODS: The study comprised 62 eyes of 35 patients with 6-month follow-up that underwent primary or secondary treatments using PASA. Pre- and postoperative results of distance and near uncorrected visual acuity (UCVA), spherical aberration (coefficient of the Z12 Zernike polynomial), and the asphericity (Q) index were reviewed. The corresponding wavefront maps (total, low, and high order aberrations) and the corresponding point spread function and modulation transfer function (MTF) were also calculated. RESULTS: Our results show that PASA improves distance and near mean UCVA, increases negative spherical aberration and negative asphericity index, and improves the corresponding MTF. CONCLUSIONS: Pseudoaccommodative advanced surface ablation is a promising approach for the surgical correction of presbyopia with distance refractive error (myopia and hyperopia with or without astigmatism). This PASA technique could theoretically be used in non-presbyopic patients with refractive error or post cataract patients with monofocal intraocular lenses. The increase in negative spherical aberration and asphericity/eccentricity index seems to increase the depth of focus of the eye, improving the near vision and compensating the age-related lens changes. Rather than creating a multifocal cornea, PASA appears to create an improved aspheric (prolate) ablation profile. PMID- 16212288 TI - Combined flap undersurface and bed LASIK for high myopia. AB - PURPOSE: To discuss the predictability, stability, and safety of LASIK for high myopia using laser treatment on both the flap undersurface and bed (both-sided LASIK). METHODS: One hundred twenty-one eyes of 63 patients underwent flap undersurface and bed LASIK for high myopia. Postoperative uncorrected visual acuity (UCVA), refraction, and topography were evaluated up to 12 months. The safety and injury indices in both-sided LASIK were compared with that of conventional LASIK. RESULTS: In both-sided LASIK, the largest diopter correction on the undersurface of the corneal flap was -6.00 diopters (D), correlating to an ablation of 44.62 microm. The mean thickness of residual corneal stroma bed was 283.87+/-19.83 microm after the laser treatment. Postoperative UCVA was close to the preoperative best spectacle-corrected visual acuity (BSCVA). The mean BSCVA was significantly better (P<.05) than preoperatively at each time point (1 week, 1, 3, and 6 months, and 1 year) after surgery. Postoperative corneal topography was normal in all cases. The safety and injury indices for conventional LASIK in both-sided LASIK eyes were calculated by programming the excimer laser as though conventional treatments would be performed to calculate the theoretical bed ablation. The safety factor of both-sided LASIK was higher than that calculated for conventional LASIK (P<.05); the injury index was lower than that calculated for conventional LASIK (P<.05). CONCLUSIONS: Surgery on high myopia by both-sided LASIK is safe and effective. PMID- 16212289 TI - LASIK enhancement using excimer laser ablation on the back of the flap. AB - PURPOSE: To report our surgical technique for LASIK enhancement by ablating the underside of the flap. METHODS: Surgical case report of clinical outcomes in three eyes of two patients who had an initial laser ablation in the bed with subsequent enhancements on the underside of the flap. RESULTS: Following LASIK enhancement on the back of the flap, two eyes gained one line of best spectacle corrected visual acuity (BSCVA) after treatment compared to preoperatively, going from 20/15 to 20/12, and one eye maintained BSCVA at 20/20. CONCLUSIONS: Excimer laser ablation of the underside of the flap is a viable alternative to treat corneas with minimal residual tissue whether primary or enhancement procedures are needed. A surgical learning curve exists with this technique. PMID- 16212290 TI - Features of hyperopic LASIK in children. AB - PURPOSE: To evaluate the results of hyperopic LASIK in children and adolescents with accommodative strabismus and refractive amblyopia. METHODS: The procedures were conducted by standard methods of LASIK using a Technolas 217C excimer laser (Bausch & Lomb, Rochester, NY). RESULTS: LASIK resolved the accommodative components of strabismus by achieving stabilization of visual acuity and decrease (or disappearance) of amblyopia degree. All patients achieved binocular vision. CONCLUSIONS: Excimer laser correction of hyperopia and hyperopic astigmatism in children and adolescents has the potential to correct refractive errors, improve visual function in amblyopic eyes, correct accommodative strabismus, and improve binocular vision. PMID- 16212291 TI - Screening patients with the corneal navigator. AB - PURPOSE: To present a corneal topography screening device for the detection of corneal ectasias and various refractive procedures based on corneal topography patterns. METHODS: A database of corneal topography patterns were analyzed and used to "train" a neural network on nine different corneal topography patterns using nineteen corneal topography indices of corneal shape and power. RESULTS: Sample normal and corneal topographies were recognized correctly. CONCLUSIONS: The use of the corneal navigator to screen various corneal topographies aids clinical diagnosis. PMID- 16212292 TI - Retinal nerve fiber layer changes after LASIK evaluated with optical coherence tomography. AB - PURPOSE: To determine whether the increase in intraocular pressure (IOP) during LASIK suction can induce a decrease in retinal nerve fiber layer thickness assessed by optical coherence tomography (OCT). METHODS: Nineteen patients (38 eyes) were enrolled in the study. Intraocular pressure was normal at all pre- and postoperative examinations. Retinal nerve fiber layer thickness was measured using OCT-3 Stratus prior to and 1 week and 3 months after LASIK. Laser in situ keratomileusis was performed using the Bausch & Lomb Hansatome microkeratome and the NIDEK EC-5000 excimer laser. Optical coherence tomography mean retinal nerve fiber layer thickness values before and after LASIK were compared using the Student paired t test. RESULTS: Mean patient age was 27.8 years (range: 18 to 33 years). Mean preoperative spherical equivalent refractive error was -4.9 diopters (D) (range: -2.0 to -8.5 D). Mean time of microkeratome suction was 30 seconds (range: 20 to 50 seconds). Preoperatively, the mean retinal nerve fiber layer thickness obtained by OCT was 104.2+/-9.0 microm; at 1 week postoperatively the mean thickness was 101.9+/-6.9 microm, and 106.7+/-6.1 microm at 3 months postoperatively. Mean retinal nerve fiber layer thicknesses obtained by OCT were not significantly different between preoperative and 1 week and 3 months after LASIK (P > or = .05). CONCLUSIONS: Laser in situ keratomileusis performed on young myopic patients does not have a significant effect on retinal nerve fiber layer thickness determined by OCT. Further studies are required to reveal the risk of possible optic nerve or retinal nerve fiber layer damage by elevated IOP during LASIK. PMID- 16212293 TI - Five techniques for improving outcomes of hyperopic LASIK. AB - PURPOSE: To determine whether five changes to our hyperopic LASIK protocol produced better outcomes. METHODS: Five changes, both technical and surgical, were instituted for the treatment of hyperopia. These five changes were nomogram refinements accounting for accommodation, use of a 7.0-mm optical zone and a 9.5 mm transition zone, a targeted mean flap diameter of 10.5 mm, sequential interruption of the laser ablation, and cleaning of the interface. The study comprised 43 eyes undergoing LASIK for hyperopia or hyperopic astigmatism with a mean preoperative spherical equivalent refraction of +2.28 diopters (D), a mean sphere of +1.93 D (range: +4.25 to +0.25 D), and mean cylinder of +1.10 D (range: +4.50 to +0.25 D). The NIDEK CXII excimer laser was used for all treatments. RESULTS: From 1 day to 3 months postoperatively, the mean hyperopic shift was <0.50 D. Postoperatively, the number of patients achieving a distance best spectacle-corrected visual acuity (BSCVA) of 20/20 gradually increased to 93% by 3 months. The number of eyes that achieved 20/15 increased by 11.7% compared to preoperatively. Eighty-eight percent of eyes maintained or gained lines of BSCVA. Four patients lost 1 line of BSCVA. One patient lost >1 line of vision due to visually significant microstriae. CONCLUSIONS: The outcomes support the observation that five surgical and technical modifications to the hyperopic LASIK procedure result in excellent visual quality and refractive outcomes and a low rate of regression. PMID- 16212294 TI - Efficacy and safety of LASIK in 10,052 eyes of 5081 myopic Chinese patients. AB - PURPOSE: To analyze the outcomes and incidence of postoperative complications in a large series of patients undergoing LASIK for myopia. METHODS: All 5081 patients (10,052 eyes) diagnosed with myopia at The Third Hospital of Handan, China, from September 2003 through March 2005 were studied. All study eyes underwent LASIK with 1-month follow-up. Spherical equivalent refraction, best spectacle-corrected visual acuity (BSCVA), and uncorrected visual acuity (UCVA) were measured before and after surgery and intra- and postoperative complications were recorded. RESULTS: Uncorrected visual acuity at 1-month follow-up of 9555 (95.1%) eyes reached or exceeded the preoperative BSCVA. Hemorrhage of corneal limbus during surgery occurred in 1060 (10.5%) eyes, Sands of Sahara syndrome occurred in 232 (2.3%) eyes, interface infection responsive to treatment occurred in 4 (0.04%) eyes, and epithelial ingrowth occurred in 1 (0.01%) eye. CONCLUSIONS: LASIK is a safe and effective method for the treatment of myopia. PMID- 16212295 TI - Clinical outcomes of CATz versus OPDCAT. AB - PURPOSE: To compare refractive outcomes, wavefront outcomes, and corneal asphericity indices (Q values) for patients treated with wavefront-guided or topography-guided custom ablations using the NIDEK Advanced Vision Excimer Laser System (NAVEX). METHODS: A total of 196 eyes of 98 patients underwent wavefront guided or topography-guided LASIK. A contralateral study of 28 eyes of 14 patients who underwent customized aspheric treatment zone (CATz) ablation in one eye and optical path difference customized aspheric treatment (OPDCAT) in the fellow eye comprised one part of the study. The second part of the study was a retrospective review of myopic LASIK using CATz in 84 eyes and LASIK using OPDCAT in 84 eyes. In the CATz-treated eyes, the preoperative mean spherical equivalent refraction was -4.06+/-1.69 diopters (D). In OPDCAT-treated eyes, the preoperative mean spherical equivalent refraction was -3.67+/-2.17 D. RESULTS: In the CATz-treated eyes, 95% of eyes achieved > or = 20/20 best spectacle-corrected visual acuity (BSCVA) with 15% gaining lines of BSCVA. In the OPDCAT-treated eyes, 94.5% of eyes achieved > or = 20/20 vision with 4.5% gaining lines of BSCVA. The difference in lines gained between CATz and OPDCAT in the retrospective component was statistically significant (P<.005). OPDCAT-treated eyes showed a statistically significant worsening in OPD root-mean-square (OPD RMS), higher order wavefront error, and corneal asphericity (P<.005) compared to preoperative in the retrospective portion of the study. In the contralateral arm of the study, OPD-RMS value was significantly higher (P<.005) in the OPDCAT eyes than the fellow CATz-treated eyes. OPDCAT-treated eyes show a larger change in the Strehl ratio compared to the CATz-treated eyes (P<.005). CONCLUSIONS: The refractive outcomes were excellent and showed no clinically significant difference between CATz and OPDCAT treatments. The higher induction of aberrations with the OPDCAT ablations may be due to the fact that these treatments are based on Zernike polynomials to drive the ablation. PMID- 16212296 TI - Wavefront-guided LASIK with the NIDEK NAVEX platform for the correction of myopia and myopic astigmatism with 6-month follow-up. AB - PURPOSE: To assess safety, efficacy, predictability, stability, and change in higher order aberrations after wavefront-guided LASIK for myopia and myopic astigmatism using the NIDEK NAVEX platform. METHODS: Wavefront-guided LASIK was performed in 93 eyes in a 6-month trial with a goal of emmetropia. Treated eyes had a mean subjective manifest spherical equivalent refraction of -4.08+/-1.99 D diopters (D), with a range of -9.50 to -0.38 D of myopia and -4.50 to 0.00 D of astigmatism. An early nomogram with the OPDCAT software program provided by the manufacturer was used in all procedures. Safety, efficacy, predictability, stability, and change in higher order aberrations were evaluated at 6 months. RESULTS: At 6 months, 100% eyes were within +/-1.0 D of emmetropia and 95% eyes were within +/-0.5 D. Uncorrected visual acuity (UCVA) of > or = 1.0 was achieved in 89% of eyes, and 38% of eyes achieved UVCA of > or = 1.2. No eyes lost > or = 2 lines of best spectacle-corrected visual acuity (BSCVA), 25% of eyes gained 1 line, 3% gained 2 lines, and 1% gained > or = 2 lines of BSCVA. Higher order root mean-square (RMS) values increased by 19% on average between pre- and postoperative measurements. Eyes treated with higher order aberrations of < 0.3 RMS showed on average an increase of 40% on preoperative values, whereas eyes with significant aberrations showed a decrease in aberrations following wavefront treatment. CONCLUSIONS: Wavefront-guided ablation using the NIDEK NAVEX platform is safe, effective, and predictable. However, patients with low amounts of aberrations showed an increase in aberrations following wavefront treatment, therefore, wavefront treatment may not be indicated or beneficial to every patient. PMID- 16212297 TI - Comparison of NIDEK CATz wavefront-guided LASIK to traditional LASIK with the NIDEK CXII excimer laser in myopia. AB - PURPOSE: To compare the visual and refractive outcomes in myopic LASIK between wavefront-guided and traditional treatment. METHODS: The study included 162 eyes of 81 myopic patients undergoing LASIK by one surgeon. The right eye of each patient underwent customized LASIK with the NIDEK NAVEX laser system (NIDEK, Gamagori, Japan) (study group); the left eye of each patient underwent traditional LASIK with the NIDEK CXII excimer laser system (control group). RESULTS: No significant differences were noted in refractive error, uncorrected visual acuity, and best spectacle-corrected visual acuity after surgery between the two groups (t test, P>.05). Postoperatively, 3.7% of patients in the study group reported glare, which was significantly lower than 12.4% in the control group (chi-square test, .01Ge=C(PPh2=NSiMe3)2] (1) with CpMn(CO)2(THF) (Cp = eta5-C5H5) in THF afforded [(Me3SiN=PPh2)2C=Ge- >Mn(CO)2Cp] (2). Similar reaction of 1 with (cod)RhCl (cod = 1,5-cyclooctadiene) in THF gave [(Me3SiN=NPPh2)2{(cod)Rh}C-GeCl] (3). The results suggested that reactive germavinylidene may exist in solution. The X-ray structures of 2 and 3 have been determined. PMID- 16212346 TI - Novel structural and magnetic properties of a 1-D iron(II)-manganese(II) LIESST compound bridged by cyanide. AB - The 1-D compound [Fe(L)(CN)2][Mn(hfac)2] (1), which adopts the -NC-Fe-CN-Mn- heterometallic structure, has been shown to exhibit light-induced excited spin state trapping effects. After illumination, anti-ferromagnetic coupling was observed between the iron(II) (S = 2) and manganese(II) (S = 5/2) ions. PMID- 16212347 TI - {Zn3(Et2O)2[(EtO)PO2(C6H5NH)]6.2THF}: a trinuclear zinc amidophosphate with an hourglass structure. AB - The reaction of ZnMe2 and the N-substituted phosphoramidic monoester [Et2NH2][(EtO)PO2(C6H5NH)] produces the trinuclear zinc cluster Zn(3)(Et2O)2[(EtO)PO2(C6H5NH)]6.2THF, demonstrating that the P-N bond can survive under mild solvothermal reaction conditions. PMID- 16212348 TI - Spin-crossover cobalt(II) compound with banana-shaped structure. AB - A banana-shaped spin-crossover (SCO) cobalt(II) complex [Co(C16-terpy)2](BPh4)2 (1) with long alkyl chains, based on a terpyridine frame, was synthesized. Compound 1 exhibited very gradual SCO behavior and changes in the dielectric constant. This shows a way in which SCO materials can be used in electronic devices. PMID- 16212349 TI - Novel 3-D framework nickel(II) complex with azide, nicotinic acid, and nicotinate(1-) as coligands: hydrothermal synthesis, structure, and magnetic properties. AB - The hydrothermal reaction of Ni(NO3)2, NaN3, and nicotinic acid (Hnic) yielded a 3-D supramolecular framework complex, [Ni(1.5)(N3)(nic)2(Hnic)]n (1) with unusual magnetic properties. In 1, Hnic and nic-1 coexist and show different coordination modes (mu2-Hnic-N,O, mu2-nic-N,O, and mu3-nic-N,O,O bridging NiII ions), and azide groups adopt mu-1,1 (EO) bridging mode linking NiII ions with an unusual Ni N-Ni angle. PMID- 16212350 TI - Cuprophilicity-induced cocrystallization of [Cu2(4,4'-bpy)(CN)2]n sheets and [Cu(SCN)]n chains into a 3-D pseudopolyrotaxane. AB - The cocrystallization of [Cu(SCN)]n chains and [Cu2(4,4'-bpy)(CN)2]n (4,4'-bpy = 4,4'-bipyridine) layers generated a 3-D pseudopolyrotaxane compound, [Cu2(4,4' bpy)(CN)2].[Cu(SCN)] (1), which can also be viewed as a 3-D network constructed by linkages of 2-D sheets and 1-D chains via unsupported CuI-CuI interactions. The CuI-CuI contact of 2.651(4) A in 1 is the shortest unsupported CuI-CuI distance documented to date, indicating cuprophilic attractions. PMID- 16212351 TI - Imidomolybdenum(IV) porphyrin complexes: synthesis, characterization, and intermetal imido transfer reactivity. AB - The imido(meso-tetra-p-tolylporphyrinato)molybdenum(IV) complexes, (TTP)Mo=NR, where R = C6H5 (1a), p-CH3C6H4 (1b), 2,4,6-(CH3)3C6H2 (1c), and 2,6-(i-Pr)2C6H4 (1d), can be prepared by the reaction of (TTP)MoCl2 with 2 equiv of LiNHR in toluene. Upon treatment of the imido complexes with pyridine derivatives, NC5H4-p X (X = CH3, CH(CH3)2, C[triple bond]N), new six-coordinate complexes, (TTP)Mo=NR.NC5H4-p-X, were observed. The reaction between the molybdenum imido complexes, (TTP)Mo=NC6H5 or (TTP)Mo=NC6H4CH3, and (TTP)Ti(eta2-PhC[triple bond]CPh) resulted in complete imido group transfer and two-electron redox of the metal centers to give (TTP)Mo(eta2-PhC[triple bond]CPh) and (TTP)Ti=NC6H5 or (TTP)Ti=NC6H4CH3. PMID- 16212352 TI - Trans-fused macrobicycle "betweenanene" around a planar Cu2(mu-CH3O)2 core showing remarkable antiferromagnetic interaction. AB - Complexation of 4,6-bis{3-(2-pyridyl)-1H-pyrazol-1-yl}pyrimidine (bppp) and copper(II) tetrafluoroborate in methanol gave a self-assembled [Cu2(CH3O)2(bppp)2]2+ ion having a betweenanene-type structure. Each bppp spans across the common Cu2O2 plane in a trans fashion to ligate each copper center. The antiferromagnetic interaction observed in the Cu2O2 core is ascribable to the superexchange along the Cu-O-Cu linkage and found to be one of the largest among the analogous compounds, owing to the large Cu-O-Cu angle perturbed with the two bppp molecular tweezers. PMID- 16212353 TI - Preference for nitrogen versus oxygen donor coordination in uranyl- and neptunyl(VI) complexes. AB - The first actinyl phosphinimine complexes have been synthesized and, in the case of uranium, exhibit strong U-N interactions. Competition reactions clearly demonstrate a surprising preference for R3P=NH ligands over R3P=O in the system [AnO2Cl2(R3PX)2] (An = U(VI), Np(VI); R = Ph, Cy; X = O, NH). Spectroscopic evidence for N-donor coordination to [NpO2]2+ in solution indicates chemical similarities to the [UO2]2+ moiety. PMID- 16212354 TI - Inclusion of a Cu2+ ion by a large-cavity crown ether dibenzo-24-crown-8 through supramolecular interactions. AB - A supramolecular copper-aqua-crown ether complex, [Cu(II)(H2O)4(dibenzo-24-crown 8]2+ (1c) is stabilized with a Lindqvist-type polyoxometalate anion, [Mo(VI)6O19]2- (1a), in an ion-pair compound [Cu(II)(H2O)4(dibenzo-24-crown 8][Mo(VI6O19] identical with [1c][1a] identical with 1. In the crystal, 1c and 1a assemble to a chainlike structure in which each polyoxoanion 1a is sandwiched by two 1c cations. 1c is a structurally characterized dibenzo-24-crown-8 (a larger cavity crown ether) supramolecular complex that shows encapsulation of a small cation at the center of its internal cavity, and compound 1 represents a unique example of a first-row transition metal-crown ether inclusion complex that interacts with a polyoxometalate anion. PMID- 16212355 TI - Niobium and tantalum tris(methimazolyl)borate complexes [M(=NC6H3iPr2 2,6)Cl2{HB(mt)3}] (M = Nb, Ta; mt = methimazolyl). AB - The first early transition metal tris(methimazolyl)borate com-plexes [M(=NR)Cl2{HB(mt)3}] (M = Nb, Ta; R = C6H3(i)Pr(2)-2,6; mt = methimazolyl) have been obtained from the reactions of [Nb(=NR)Cl3(DME)] or [Ta(=NR)Cl3(THF)2] (DME = dimethyl ether; THF = tetrahydrofuran) with Na[HB(mt)3] and structurally characterized, illustrating that the HB(mt)3 ligand can indeed be compatible with "hard" metals in high oxidation states. PMID- 16212356 TI - Heterobimetallic reductive cross-coupling of benzonitrile with carbon dioxide, pyridine, and benzophenone. AB - Described herein are heterobimetallic radical cross-coupling reactions between the benzonitrile adduct of the molybdenum(III) complex Mo(N[t-Bu]Ar)3 (Ar = 3,5 C6H3Me2) and titanium(III) complexes with carbon dioxide, pyridine, and benzophenone. The titanium(III) system employed was either Ti(N[t-Bu]Ar)3 (Ar = 3,5-C6H3Me2) or Ti(N[t-Bu]Ph)3. Crystal structure studies are described for the Mo/PhCN/CO2/Ti coupled system and for an analogue of the Mo/PhCN/Ph2CO/Ti coupled system in which PhCN is replaced with 2,6-Me2C6H3CN. In the case of the couplings involving pyridine and benzophenone, C-C bond formation takes place with dearomatization, with the new C-C bond being formed between the nitrile carbon of PhCN and the para carbon of pyridine or one of the benzophenone phenyl groups. Of the radical metal complex/substrate adducts invoked in this work, that between titanium(III) and CO2 is the only one not directly observable. In all cases, the selective cross-coupling reactions are interpreted as arising by heterodimerization of titanium(III) substrate complexes (substrate = CO2, py, or Ph2CO) with the persistent molybdenum-PhCN radical adduct. All of the heterobimetallic coupling products are diamagnetic, and the metal ions Ti and Mo in them both are assigned to the formal 4+ oxidation state. PMID- 16212357 TI - Fluorinated tris(pyrazolyl)borate ligands without the problematic hydride moiety: isolation of copper(I) ethylene and copper(I)-tin(II) complexes using [MeB(3 (CF3)Pz)3]-. AB - The thallium derivative of a fluorinated, B-methylated, tris(pyrazolyl)borate ligand, [MeB(3-(CF3)Pz)3]-, has been synthesized via a two-step process using the corresponding pyrazole, Li[MeBH3], and thallium(I) acetate. Reaction of [MeB(3 (CF3)Pz)3]Tl with CuBr in the presence of ethylene leads to [MeB(3 (CF3)Pz)3]Cu(C2H4). It is a thermally stable solid. [MeB(3-(CF3)Pz)3]Cu(C2H4) reacts with [(Bn)2ATI]SnCl to yield [MeB(3-(CF3)Pz)3]Cu<--Sn(Cl)[(Bn)2ATI], featuring an unsupported Cu(I)-Sn(II) bond [2.4540(4) A]. PMID- 16212358 TI - Polymer building blocks: self-assembly of silver(I) cyclotriphosphazene cationic columns. AB - The multimodal ligand hexakis(2-pyridyloxy)cyclotriphosphazene (L) and its 4 methyl-2-pyridyloxy analogue (MeL) react with Ag(I) to afford {[AgL]+}infinity supramolecular cationic columns via self-assembly, with the anions occupying the intercolumnar channels. In contrast, the reaction of MeL with Cu(I) yields a dimetallic Cu(II) complex containing mu-OH and mu-4-methyl-2-pyridyloxylato bridges. PMID- 16212359 TI - Chemistry of transition-metal clusters with mixed Sb/S ligands: evidence for a terminal Sb=S double bond in Cp*3Rh3Sb2S5 (Cp* = C5Me5). AB - The reaction of [Cp2*Rh2Cl4] (Cp* = C5Me5) with a slight excess of K(3)SbS(3) in boiling THF gave the neutral clusters [Cp*4Rh4S5] (1), [Cp*3Rh3Sb2S5] (2), and after salt metathesis [Cp*3Rh3SbSn]PF6 (3; n = 5 and 6). The structures of 1-3 are heterocubane clusters with CpRh, S, and Sb vertices but with sulfur inserted into one (1 and 2) or two (3) edges. X-ray diffraction analysis of 2 additionally reveals a very short Sb-S distance of 2.297(1) A within the novel mu3-Sb2S4 ligand. Density functional theory calculation of the model compounds [SSbS]3-, [HSSbS]2-, and [HSSbH2S]0 provided strong evidence for the existence of a stable terminal Sb=S double bond in 2. PMID- 16212360 TI - Electronic structures of six-coordinate ferric porphyrin complexes with weak axial ligands: usefulness of 13C NMR chemical shifts. AB - 1H NMR, (13)C NMR, and EPR spectra of six-coordinate ferric porphyrin complexes [Fe(Por)L2]ClO4 with different porphyrin structures are presented, where porphyrins (Por) are planar 5,10,15,20-tetraphenylporphyrin (TPP), ruffled 5,10,15,20-tetraisopropylporphyrin (TiPrP), and saddled 2,3,7,8,12,13,17,18 octaethyl-5,10,15,20-tetraphenylporphyrin (OETPP), and axial ligands (L) are weak oxygen ligands such as pyridine-N-oxide, substituted pyridine-N-oxide, DMSO, DMF, MeOH, THF, 2-MeTHF, and dioxane. These complexes exhibit the spin states ranging from an essentially pure high-spin (S = 5/2) to an essentially pure intermediate spin (S = 3/2) state depending on the field strength of the axial ligands and the structure of the porphyrin rings. Reed and Guiset reported that the pyrrole-H chemical shift is a good probe to determine the spin state in the spin admixed S = 5/2,3/2 complexes (Reed, C. A.; Guiset, F. J. Am. Chem. Soc. 1996, 118, 3281 3282). In this paper, we report that the chemical shifts of the alpha- and beta pyrrole carbons can also be good probes to determine the spin state because they have shown good correlation with those of the pyrrole-H or pyrrole-C(alpha). By putting the observed or assumed pyrrole-H or pyrrole-C(alpha) chemical shifts of the pure high-spin and pure intermediate-spin complexes into the correlation equations, we have estimated the carbon chemical shits of the corresponding complexes. The orbital interactions between iron(III) and porphyrin have been examined on the basis of these chemical shifts, from which we have found that both the d(xy)-a(2u) interaction in the ruffled Fe(T(i)PrP)L2+ and d(xy)-a(1u) interaction in the saddled Fe(OETPP)L2+ are quite weak in the high-spin and probably in the intermediate-spin complexes as well. Close inspection of the correlation lines has suggested that the electron configuration of an essentially pure intermediate-spin Fe(T(i)PrP)L2+ changes from (d(xy), d(yz))3(d(xy))1(d(z)2)1 to (d(xy))2(d(xz), d(yz))2(d(z)2)1 as the axial ligand (L) changes from DMF to MeOH, THF, 2-MeTHF, and then to dioxane. Although the DFT calculation has indicated that the highly saddled intermediate-spin Fe(OETPP)(THF)2+ should adopt (d(xy), d(yz))3(d(xy))1(d(z)2)1 rather than (d(xy))2(d(xz), d(yz))2(d(z)2)1 because of the strong d(xy)-a(1u) interaction (Cheng, R.-J.; Wang, Y.-K.; Chen, P.-Y.; Han, Y.-P.; Chang, C.-C. Chem. Commun. 2005, 1312-1314), our 13C NMR study again suggests that Fe(OETPP)(THF)2+ should be represented as (d(xy))2(d(xz), d(yz))2(d(z)2)1 because of the weak d(xy)-a(1u) interaction. The contribution of the S = 3/2 state in all types of the spin admixed S = 5/2,3/2 six-coordinate complexes has been determined on the basis of the (13)C NMR chemical shifts. PMID- 16212361 TI - Bis(mu-oxo)dicopper(III) complexes of a homologous series of simple peralkylated 1,2-diamines: steric modulation of structure, stability, and reactivity. AB - We have synthesized and characterized bis(mu-oxo)dicopper(III) dimers 1b-4b (Os) based on a core family of peralkylated trans-(1R,2R)-cyclohexanediamine (CD) ligands, self-assembled from the corresponding [LCu(MeCN)]CF3SO3 species 1a-4a and O2 at 193 K in aprotic media; additional Os based on peralkylated ethylenediamine and tridentate polyazacyclononane ligands were synthesized analogously for comparative purposes (5b-7b and 8b-9b, respectively). Trigonal planar [LCu(MeCN)]1+ species are proposed as the active O precursors. The 3 coordinate Cu(I) complexes [(L(TE))Cu(MeCN)]CF3SO3 (4a) and [(L(TB))Cu(MeCN)]CF3SO3 (10a) were structurally characterized; the apparent O2 inertness of 10a correlates with the steric demands of its four benzyl substituents. The rate of O formation, a multistep process that likely proceeds via associative formation of a 1:1 [LCu(O2)]1+ intermediate, exhibits significant dependence upon ligand sterics and solvent: oxygenation of 4a-the slowest reacting O precursor of the CD series-is first-order with respect to [4a] and proceeds at least 300 times faster in tetrahydrofuran than in CH2Cl2. The EPR, UV vis, and resonance Raman spectra of 1b-9b are all characteristic of the diamagnetic bis(mu-oxo)dicopper(III) core. The intense ligand-to-metal charge transfer absorption maxima of CD-based Os are red-shifted proportionally with increasing peripheral ligand bulk, an effect ascribed to a slight distortion of the [Cu2O2] rhomb. The well-ordered crystal structure of [(L(ME))2Cu2(mu O)2](CF3SO3)2.4CH2Cl2 ([3b. 4CH2Cl2]) features the most metrically compact [Cu2O2]2+ core among structurally characterized Os (av Cu-O 1.802(7) A; Cu...Cu 2.744(1) A) and exemplifies the minimal square-planar ligation environment necessary for stabilization of Cu(III). The reported Os are mild oxidants with moderate reactivity toward coordinating substrates, readily oxidizing thiols, certain activated alkoxides, and electron-rich phenols in a net 2e-, 2H+ process. In the absence of substrates, 1b-9b undergo thermally induced autolysis with concomitant degradation of the polyamine ligands. Ligand product distribution and primary kinetic isotope effects (kobsH/kobsD approximately 8, 1b/d24-1b, 293 K) support a unimolecular mechanism involving rate-determining C-H bond cleavage at accessible ligand N-alkyl substituents. Decomposition half-lives span almost 3 orders of magnitude at 293 K, ranging from approximately 2 s for 4b to almost 30 min for d(24)-1b, the most thermally robust dicationic O yet reported. Dealkylation is highly selective where ligand rigidity constrains accessibility; in 3b, the ethyl groups are attacked preferentially. The observed relative thermal stabilities and dealkylation selectivities of 1b-9b are correlated with NC(alpha)-H bond dissociation energies, statistical factors, ligand backbone rigidity, and ligand denticity/axial donor strength. Among the peralkylated amines surveyed, bidentate ligands with oxidatively robust NC(alpha)-H bonds provide optimal stabilization for Os. Fortuitously, the least sterically demanding N-alkyl substituent (methyl) gives rise to the most thermally stable and most physically accessible O core, retaining the potential for exogenous substrate reactivity. PMID- 16212362 TI - New palladium(II) and platinum(II) complexes with the model nucleobase 1 methylcytosine: antitumor activity and interactions with DNA. AB - Palladium and platinum complexes with the model nucleobase 1-methylcytosine (1 Mecyt) of the types [Pd(N-N)(C6F5)(1-Mecyt)]ClO4 [N-N = bis(3,5-dimethylpyrazol-1 yl)methane (bpzm), bis(pyrazol-1-yl)methane (bpzm), N,N,N',N' tetramethylethylenediamine (tmeda), or 2,2'-bipyridine (bpy)] and [M(dmba)(L')(1 Mecyt)]ClO4 [dmba = N,C-chelating 2-(dimethylaminomethyl)phenyl; L' = PPh(3) (M = Pd or Pt), DMSO (M = Pt)] have been obtained. Palladium and platinum complexes of the types cis-[M(C6F5)2(1-Mecyt)2] (M = Pd or Pt) and cis-[Pd(L')(C6F5)(1 Mecyt)2]ClO4 (L' = PPh(3) or t-BuNC) have also been prepared. The crystal structures of [Pd(bpzm)(C6F5)(1-Mecyt)]ClO4, [Pt(dmba)(DMSO)(1-Mecyt)]ClO4, cis [Pd(C6F5)2(1-Mecyt)2], and cis-[Pd(t-BuNC)(C6F5)(1-Mecyt)2]ClO4 have been established by X-ray diffraction. There is extensive hydrogen bonding (N-H...O, C H...F or C-H...O) in all the compounds. There are also intermolecular pi-pi interactions between pyrimidine rings of adjacent chains in [Pd(C6F5)2(1 Mecyt)2]. DNA adduct formation of the new complexes synthesized was followed by circular dichroism and electrophoretic mobility. Atomic force microscopy images of the modifications caused by the complexes on plasmid DNA pBR322 were also obtained. Values of IC(50) were also calculated for the new complexes against the tumor cell line HL-60. At a short incubation time (24 h) almost all new complexes were more active than cisplatin. PMID- 16212363 TI - Photophysical properties of Ru(II) bipyridyl complexes containing hemilabile phosphine-ether ligands. AB - Emission and absorbance spectra, along with low-temperature excited-state lifetimes, were obtained for the hemilabile complexes, [Ru(bpy)2L](PF6)2 [L = (2 methoxyphenyl)diphenylphosphine (RuPOMe) (1) and (2 ethoxyphenyl)diphenylphosphine (RuPOEt) (2)] in solid 4:1 ethanol/methanol solution. Spectral data were evaluated with ground-state reduction potentials using Lever parameters. Lifetime data for these complexes were collected from 77 to 160 K, and the rate constant for the combined radiative and nonradiative decay process, k, the thermally activated process prefactor, k'(0), the rate constant for the MLCT --> d-d transition, k', and the activation energy, DeltaE', were calculated from a plot of ln(1/tau) versus 1/T for both (1) and (2). The low temperature luminescence lifetimes of (1) were observed to decrease with increases in water concentration. The photophysical and kinetic data of (1) and (2) are compared to literature data for [Ru(bpy)3](PF6)2. The emission maxima of (1) and (2) are blue-shifted relative to [Ru(bpy)3](PF6)2 due to the presence of the strong-field phosphine ligand, which enhances pi back-bonding to the bipyridyl ligands. The thermal activation energy, DeltaE', is significantly larger for [Ru(bpy)3](PF6)2 than for (1) and (2) resulting in a faster MLCT --> d d transition for (1) and (2). These results are discussed in the context of radiationless decay through thermally activated ligand-field states on the metal complex. PMID- 16212364 TI - Solvothermal synthesis of nanocrystalline copper nitride from an energetically unstable copper azide precursor. AB - Nonaqueous solvothermal chemical reactions have found extensive utility in the growth of inorganic non-oxide materials. This report describes the successful use of organic solvothermal environments to synthesize energetically unstable copper azide precursors that are then decomposed in situ to crystalline metastable copper nitride at temperatures below 200 degrees C. A comparison of Cu3N products formed from nonpolar (toluene) and coordinating (THF) solvents is described. The cubic Cu3N products are nanocrystalline with aggregated particle-like extended structures and were characterized by X-ray diffraction, electron microscopy, IR spectroscopy, and mass spectrometry. The thermal stability and composition of Cu3N was examined by thermogravimetric analysis and bulk elemental analysis. The particle surfaces contain bound residual solvent species that can be removed by heating. The poorly coordinating solvent, toluene, lead to a more crystalline product containing less residual organic content. Benchtop reactions were performed to follow the temporal formation and decomposition of metal azide intermediates. These studies provided more detailed information on the progression of metal azide to metal nitride materials in a solvothermal environment. PMID- 16212365 TI - Tribasic lead maleate and lead maleate: synthesis and structural and spectroscopic characterizations. AB - We report on the synthesis and structure of tribasic lead maleate hemihydrate ([Pb4O3]C2H2(CO2)2.(1/2)H2O, TRIMAL) and lead maleate (PbC2H2(CO2)2, PBMAL). The structure of [Pb4O3]C2H2(CO2)2.(1/2)H2O, solved ab initio from X-ray powder diffraction data, consists of infinite slabs of edge-sharing OPb4 tetrahedra, of composition [Pb4O3], running along the c axis and linked together into a three dimensional network by tetradentate maleate anionic ligands. The structure of PbC2H2(CO2)2, solved from single crystal diffraction data, is lamellar and contains double layers of heptacoordinated lead atoms, bonded only to the oxygen atoms of the maleate ligands. In both compounds, lead is in the oxidation state 2+ and the coordination polyhedra around the Pb2+ exhibit a hemidirected geometry and are strongly distorted as a result of the lone pair of electrons. The absence of protons on the acidic portion of the maleate moieties was confirmed by Raman spectroscopy and by 1H MAS and 1H-13C CP MAS NMR experiments. The two compounds were further characterized using chemical and thermogravimetric analyses. PMID- 16212366 TI - Uranium(III)/(IV) nitrile adducts including UI4(N[triple bond]CPh)4, a synthetically useful uranium(IV) complex. AB - The synthesis of complexes used to elucidate an understanding of fundamental An(III) and An(IV) coordination chemistry requires the development of suitable organic-soluble precursors. The reaction of oxide-free uranium metal turnings with 1.3 equivalents of elemental iodine in acetonitrile provided the U(III)/U(IV) complex salt, [U(N[triple bond]CMe)9][UI6][I] (1), in which the U(III) cation is surrounded by nine acetonitrile molecules in a tricapped trigonal prismatic arrangement, a [UI6]2- counterion, and a noncoordinating iodide. The U-N distances for the prismatic and capping nitrogens are 2.55(3) and 2.71(5) A, respectively. The same reaction performed in benzonitrile afforded crystalline UI4(N[triple bond]CPh)4 (3) in 78% isolated yield. In the solid state, 3 shows an eight-coordinate U(IV) atom in a "puckered" square antiprismatic geometry with U-N and U-I distances of 2.56(1) and 3.027(1) A, respectively. This benzonitrile UI4 adduct is a versatile U(IV) synthon that is soluble in methylene chloride, benzonitrile, and tetrahydrofuran, and moderately soluble in toluene and benzene, but decomposes in benzonitrile at 198 degrees C to [UI(N[triple bond]CPh)8][UI]6 (4), a U(III)/U(IV) salt analogous to 1. A toluene slurry of 3 treated with 2.2 equiv of Cp*MgCl.THF (Cp* = pentamethylcyclopentadienide) provided Cp*2UI2(N[triple bond]CPh) (5) in low yields. Single-crystal X-ray structure determination shows that the iodide ligands in 5 are in a rare cis configuration with an acute I-U-I angle of 83.16(7) degrees . Treatment of a methylene chloride solution of 3 with KTp* (Tp* = hydridotris(3,5-dimethylpyrazolylborate)) formed green TpUI3 (6) which was converted to yellow Tp*UI3(N[triple bond]CMe) (7) by rinsing with acetonitrile. Addition of 2.2 equiv of KTp* to a toluene solution of 3 followed by heating at 95 degrees C, filtration, and crystallization led to the isolation of the dinuclear species [Tp*UI(dmpz)]2[mu-O] (9) (dmpz = 3,5-dimethylpyrazolide), presumably formed by hydrolytic cleavage of excess KTp* by adventitious water. The Tp* complexes 6, 7, and 9 were characterized by single-crystal X-ray diffraction, NMR, FT-IR, and optical absorbance spectroscopies. PMID- 16212367 TI - Influence of ligand rigidity and ring substitution on the structural and electronic behavior of trivalent iron and gallium complexes with asymmetric tridentate ligands. AB - Species 1-6 are [M(III)(L)2]ClO4 complexes formed with the PhO--CH=N-CH2-Py imines, (L(I))- and (L(tBuI))-, and PhO--CH2-NH-CH2-Py amines, (L(A))- and (L(tBuA))-, in which PhO- is a phenolate ring and Py is a pyridine ring and the prefix tBu indicates the presence of tertiary butyl groups occupying the positions 4 and 6 of the phenol ring. Monometallic species with d5 high-spin iron (1, 2, 3, 4) and d10 gallium (5, 6) were synthesized and characterized to assess the influence of the ligand rigidity and the presence of tertiary butyl substituted phenol rings on their steric, electronic, and redox behavior. Characterization by elemental analysis, mass spectrometry, IR, UV-visible, and EPR spectroscopies, and electrochemistry has been performed, and complexes [FeIII(L(tBuI))2]ClO4 (2), [FeIII(L(tBuA))2]ClO4 (4), and [Ga(III)(L(tBuI))2]ClO4 (5) have been characterized by X-ray crystallography. The crystal structures show the imine ligands meridionally coordinated to the metal centers, whereas the amine ligands are coordinate in a facial mode. Cyclic voltammetry shows that the complexes with the ligands (L(tBuI))- and (L(tBuA))- were able to generate ligand based phenoxyl radicals, whereas unsubstituted ligands displayed ill-defined redox processes. EPR spectroscopy supports high-spin configurations for the iron complexes. UV-visible spectra are dominated by charge-transfer phenomena, and imine compounds exhibit dramatic hyperchromism when compared to equivalent amines. The tertiary butyl groups on the phenolate ring enhance this trend. Detailed B3LYP/6-31G(d)-level calculations have been used to account for the results observed. PMID- 16212368 TI - KBi(2-x)Pbx (0 < x or = 0.6, these tetrahedra become alternately elongated and contracted. The distortion of the framework lowers the space-group symmetry to F43m (KBi(1.2)Pb(0.8), F43m, Z = 8, a = 9.572(1) A). Magnetometer measurements show that KBi2 (x = 0) is metallic and goes through a superconducting transition below 3.5 K. First principles calculations reveal that the Fd3m --> F43m distortion is largest for KBiPb (x = 1.0), which at the same time turns into a semiconductor. Thus, F43m KBiPb corresponds to a proper charge-balanced Zintl phase, K+[BiPb]-, with separated polyanionic tetrahedra, (Bi2Pb2)2-. However, it was not possible to prepare F43m KBiPb. Syntheses attempting to increase the Pb content in KBi(2-x)Pbx above x = 0.8 yielded additional, not yet characterized, ternary phases. PMID- 16212369 TI - Structural and spectroscopic study of reactions between chelating zinc-binding groups and mimics of the matrix metalloproteinase and disintegrin metalloprotease catalytic sites: the coordination chemistry of metalloprotease inhibition. AB - To understand the coordination chemistry of zinc-binding groups (ZBGs) with catalytic zinc centers in matrix metalloproteinases (MMPs) and disintegrin metalloproteases (ADAMs), we have undertaken a model compound study centered around tris(3,5-methylphenypyrazolyl)hydroboratozinc(II) hydroxide and aqua complexes ([Tp(Ph,Me)ZnOH] and [Tp(Ph,Me)Zn(OH2)]+, respectively, wherein (Tp(Ph,Me))- = hydrotris(3,5-methylphenylpyrazolyl)borate) and the products of their reactions with a class of chelating Schiff's base ligands. The results show that the protic ligands, HL (HL = N-propyl-1-(5-methyl-2-imidazolyl)methanimine (5-Me-4-ImHPr), N-propyl-1-(4-imidazolyl)methanimine (4-ImHPr), and N-propyl-1-(2 imidazolyl)methanimine (2-ImHPr)), react with [Tp(Ph,Me)ZnOH] and give products with the general formula [Tp(Ph,Me)ZnL], whereas reactions with neutral aprotic ligands, L' (L' = N-propyl-1-(1-methyl-2-imidazolyl)methanimine (1-Me-2-ImPr) and N-propyl-1-(2-thiazolyl)methanimine (2-TaPr)), yield the corresponding [Tp(Ph,Me)ZnL]+ complexes. Although the phenol group of N-propyl-1-(2 hydroxyphenyl)methanimine (2-HOPhPr) is protic, this ligand forms a cationic four coordinate complex containing an intraligand hydrogen bond. The solid-state structures of these complexes were determined by single-crystal X-ray diffraction, and the results showed that the protic ligands form five-membered chelates of the Zn2+ ion. All ligands displace the aqua ligand in [Tp(Ph,Me)Zn(OH2)]+ to yield complexes having 1H NMR spectra consistent with the formation of five membered chelates. The 1H resonance frequencies of the chelating ligands typically shift upfield upon coordination to the zinc center, due to ring current effects from the pendant phenyl groups of the (Tp(Ph,Me))- ligand. Thus, the 1H NMR spectra provide a convenient and sensitive means of tracking the solution reactions by titration. The resulting series of spectra showed that the stabilities of the chelates in solution depend on the propensity of the ligands to deprotonate upon chelation of the zinc center. The behaviors of these bidentate ZBGs provide insight into the structural and electronic factors that contribute to the stabilities of inhibited MMPs and ADAMs and suggest that the proton acidity of the coordinated ZBG may be a crucial criterion for inhibitor design. PMID- 16212370 TI - Evidence of incipient bond-stretching isomerism in Sr2.04(1)Ca0.96(1)Sn5 from variable-temperature structural studies. AB - The title compound was found to crystallize in the Pu3Pd5 structure type (SG Cmcm) with cell dimensions of a = 10.5179(9) A, b = 8.4789(8) A, and c = 10.7623(10) A. The structure consists of isolated Sn5(6-) square-pyramidal units surrounded by cations that seem to play a crucial role in stabilizing the Zintl polyanions. The square pyramids contract at low temperatures (100 K) leading to the shortening of the basal intracluster Sn-Sn bond (2.74 A), while the intercluster bonds become very large, indicating features of bond stretching isomerism as is known for Ba3Ge4. A study of different crystals shows a slight variation in the lattice parameters, suggesting the presence of a definite phase width which was substantiated by the successful synthesis of monophasic samples of Sr(3-x)CaxSn5 (0.5 1 unit lower than for related cis complexes. PMID- 16212374 TI - Trilacunary heteropolytungstates functionalized by organometallic ruthenium(II), [(RuC6H6)2XW9O34]6- (X = Si, Ge). AB - The benzene-Ru(II)-supported trilacunary heteropolytungstates [(RuC6H6)2XW9O34]6- (X = Si, 1; Ge, 2) have been synthesized and characterized by multinuclear solution NMR (183W, 13C, 1H, 29Si), UV-vis and IR spectroscopy, electrochemistry, and elemental analysis. Single-crystal X-ray analysis was carried out on Rb2Na4[(RuC6H6)2SiW9O34].21H2O (RbNa-1), which crystallizes in the triclinic system, space group P, with a = 11.9415(2) A, b = 13.3123(2) A, c = 19.4927(4) A, alpha = 96.6460(10) degrees , beta = 95.1570(10) degrees , gamma = 98.2560(10) degrees , and Z = 2 and on Cs2Na4[(RuC6H6)2GeW9O34].19.5H2O (CsNa-2), which crystallizes also in the triclinic system, space group P, with a = 11.930(4) A, b = 13.353(4) A, c = 19.586(6) A, alpha = 95.982(5) degrees , beta = 95.414(6) degrees , gamma = 98.142(5) degrees , and Z = 2. The novel polyanion structure consists of two (RuC6H6) units linked to a trilacunary (XW9O34) Keggin fragment via Ru-O(W) and Ru-O(X) bonds resulting in an assembly with Cs symmetry. Polyanions 1 and 2 were synthesized by reaction of [RuC6H6Cl2]2 with [A-alpha XW9O34]10- in aqueous buffer medium (pH 6.0). Both 1 and 2 are stable in solution as indicated by the expected 5-line pattern (2:1:2:2:2) in the 183W NMR and the expected (3C, 1H, and 29Si spectra. Descriptions of the respective electrochemical behaviors of the W centers and the Ru centers in 1 and 2 are given in media where these processes are clearly defined. In a pH = 3 acetate medium, the cyclic voltammetry of the W centers shows the known fingerprint of the trilacunary alpha-[XW9O34]n- (X = Ge, Si) moieties. The presence of the (RuC6H6) substituents imparts a good stability to these fragments in solution. Stepwise oxidation of the Ru centers was suspected in pH = 5 acetate medium, but only the first step was well-separated from a large current composite wave. The stepwise oxidation was finally observed clearly in a DMF-water (90/10 v/v) mixture and shows two well-behaved Ru oxidation processes. A short comparison is made with DMSO-bearing Ru polyoxometalates. PMID- 16212375 TI - Oxygen atom transfer reactivity from a dioxo-Mo(VI) complex to tertiary phosphines: synthesis, characterization, and structure of phosphoryl intermediate complexes. AB - The oxygen atom transfer (OAT) reactivity of TpMoO2Cl with PMe3, PEt3, and PPhMe2 (where Tp = hydrotris(3,5-dimethylpyrazol-1-yl)borate) has been investigated. The OAT reactions proceed through a diamagnetic Mo(IV) phosphoryl intermediate complex of general formula TpMoOCl(OPR3) (OPR3 = OPMe3, OPEt3, OPPhMe2), which have been isolated and characterized by 1H and 31P NMR, UV-visible, and infrared spectroscopies and electrospray ionization mass spectrometry. Solid-state crystal structures of TpMoOCl(OPMe3) and TpMoOCl(OPPhMe2) are also reported, the oxygen to-phosphorus distances agree with a double-bond formulation and a single bond between the metal and the phosphoryl oxygen atom. The stability of the phosphoryl intermediate complexes depends on the steric properties of the coordinated phosphine-oxides. These intermediate complexes have been converted to solvent coordinated species, TpMoOCl(solv) (solv = acetonitrile or dmf), and the coordinated solvents exchange with the bulk solvent. PMID- 16212376 TI - Role of anions on the crystal structures of copper(II) and zinc(II) complexes of a tunable butterfly cyclophane macrocycle. AB - Three crystal structures of a ditopic cyclophane ligand (L) in which two 1,5,8,12 tetraamine molecules have been attached through methylene spacers to the ortho positions of a benzene ring are reported. The first one (1) corresponds to the tetraprotonated free macrocycle (H4L4+) having two tetrachlorozincate(II) counteranions (C24H54O2N8Cl8Zn2, a = 9.1890(2) A, b = 14.0120(3) A, c = 15.3180(3) A, alpha = 89.2320(7) degrees , beta = 82.0740(6) degrees , gamma = 83.017(1) degrees , Z = 2.00, triclinic, P); the second one (2) is of a binuclear Cu2+ complex having coordinated chloride anions and perchlorate counteranions (C24H58O14N8Cl4Cu2 a = 9.9380(2) A, b = 30.2470(6) A, c = 53.143(1) A, orthorhombic, F2dd, Z = 18), and the third one (3) corresponds to an analogous Zn2+ complex that has been crystallized using triflate as counteranion (C26H(51.2)O(6.6)N8Cl2F6S2Zn2 a = 8.472(5) A, b = 9.310(5), c = 13.745(5) A, alpha = 84.262(5) degrees , beta = 77.490(5) degrees , gamma = 73.557(5) degrees , triclinic, P, Z = 2). The analysis of the crystallographic data clearly shows that the conformation of the macrocycle and, in consequence, the overall architecture of the crystals are controlled by the anions present in the moiety, pi-pi-stacking associations, and hydrogen bonding interactions. The protonation and stability constants for the formation of the Cu2+ and Zn2+ complexes in aqueous solution have been determined potentiometrically in 0.15 mol dm(-3) NaClO4 at 298.1 K. Intramolecular hydrogen bonding defines the protonation behavior of the compound. Positive cooperativity is observed in the formation of the Cu2+ complexes. PMID- 16212377 TI - NMR investigation of the interaction of vanadate with carbasilatranes in aqueous solutions. AB - Reaction of vanadate with carbasilatranes [methoxy{N,N',N' '-2,2',3-[bis(1 methylethanolato)(propyl)]amino}silane (1), methoxy{N,N',N' '-2,2',3-[bis(1 ethanolethanolato)(propyl)]amino}silane (2), and {N,N',N' '-2,2',2 [bis(ethanolato)(glycolpropyl ether)]amino}silane (3)] in aqueous solution results in the formation of vanadosilicates and five-coordinated chelate vanadium(V) complexes as evidenced by 51V, 1H, and 13C NMR spectroscopy. Chiral carbasilatrane S,S-1 was characterized in the solid state by X-ray diffraction, revealing a trigonal bipyramidal geometry around the metal ion, with one unidentate methoxy group and one atrane nitrogen atom at the axial positions and one carbon and two atrane oxygen atoms at the equatorial plane of the bipyramid. Crystal data (Mo Kalpha; 100(2) K) are as follows: orthorhombic space group P2(1)2(1)2(1); a = 8.8751(6), b = 9.7031(7), c = 14.2263(12) A; Z = 4. The complexation of vanadium either with 1 or 2 is stereoselective yielding approximately 94% of the complex containing ligand in the S,R-configuration. The lower ability of the S,S- and R,R-diastereoisomers of 1 and 2 to ligate vanadate was attributed to stereochemical factors, dictating a square pyramidal geometry for the chelated complexes. A dynamic process between the vanadium chelate complexes and the respective carbasilatranes was evaluated by 2D {1H} EXSY NMR spectroscopy. These spectra show that the vanadate complexes with the open carbasilatranes exchange more slowly with the free ligand compared to the respective alcohol aminate complexes. PMID- 16212378 TI - Magnetic properties of isostructural BaCoP2O7, BaNiP2O7, and BaCuP2O7 studied with dc and ac magnetization and specific heat. AB - Magnetic properties of three isostructural compounds BaMP2O7 (M = Co, Ni, and Cu) were investigated by dc and ac magnetization and specific heat measurements. BaCuP2O7 was shown to be an excellent quasi-one-dimensional linear-chain Heisenberg antiferromagnet with an exchange constant (J/kB) of 103.8 K (Hamiltonian H = J Sigma SiS(i+1)) and a temperature for the long-range magnetic order (TN) of 0.81 K giving the ratio kBTN/J = 0.78%. BaCoP2O7 and BaNiP2O7 exhibited long-range antiferromagnetic order at TN = 10.4 and 10.1 K, respectively. BaCoP2O7 and BaNiP2O7 showed a large contribution of the short range correlation above TN. BaNiP2O7 remained in the antiferromagnetic state up to 90 kOe at 2 K, whereas BaCoP2O7 demonstrated two metamagnetic phase transitions at about 52 and 71 kOe at 2 K if the magnetic field was parallel to the easy direction. BaMP2O7 melted incongruently at 1323 K (M = Co), 1344 K (M = Ni), and 1338 K (M = Cu). PMID- 16212379 TI - Structure and magnetic properties of (meso-tetraphenylporphinato)manganese(III) bis(dithiolato)nickelates. AB - The crystal structures of [MnTPP]{Ni[S2C2H(CN)]2} [MnTPP = (meso tetraphenylporphinato)manganese(III)] and [MnTPP]{Ni[S2C2(CN)2]2} have been determined. These salts possess trans-mu-coordination of S = 1/2 {Ni[S2C2H(CN)]2}*- and {Ni[S(2)C(2)(CN)(2)](2)}*- to Mn(III) and form parallel 1 D coordination polymer chains exhibiting nu(CN) at 2210 and 2200 and 2220 and 2212 cm(-1), respectively. The bis(dithiolato) monoanions are planar and bridge two cations with MnN distances of 2.339(16), and 2.394(3) A, respectively, which are comparable to related MnN distances observed for [MnTPP][TCNE].x(solvates). In addition, [MnTP'P]{Ni[S2C2(CN)2]2} {H2TP'P = meso-tetrakis[3,5-di-tert-butyl-4 hydroxyphenyl)porphyrin] and [MnTP'P(OH2)]{Ni[S2C2(CN)2]2} were prepared. The latter forms isolated paramagnetic ions. The room-temperature values of chiT for 1-D [MnTPP]{Ni[S2C2H(CN)]2}, [MnTPP]{Ni[S2C2(CN)2]2}, and [MnTP'P]{Ni[S2C2(CN)2]2} are 2.55, 3.28, and 2.86 emu K/mol, respectively. Susceptibility (chi) measurements between 2 and 300 K reveal weak antiferromagnetic interactions with theta= -5.9 and -0.2 K for [MnTPP]{Ni[S(2)C(2)H(CN)](2)} and [MnTPP]{Ni[S2C2(CN)2]2}, respectively, and stronger antiferromagnetic coupling of -50 K for [MnTP'P]{Ni[S2C2(CN)2]2} from fits of chi(T) to the Curie-Weiss law. The 1-D intrachain coupling, J(intra), of [MnTPP]{Ni[S2C2H(CN)]2} and [MnTPP]{Ni[S2C2(CN)2]2} was determined from modeling chiT(T) by the Seiden expression (H = -2JSi.Sj) with J/kB = -8.00 K (-5.55 cm( 1); -0.65 meV) for [MnTPP]{Ni[S2C2H(CN)]2}, J/kB = -3.00 K (-2.08 cm(-1); -0.25 meV) for [MnTP'P]{Ni[S2C2(CN)2]2}, and J/kB = -122 K (-85 cm(-1)) for [MnTP'P]{Ni[S2C2(CN)2]2}. These observed negative J(intra)/kB values are indicative of antiferromagnetic coupling. These materials order as ferrimagnets at 5.5, 2.3, and 8.0 K, for [MnTPP]{Ni[S2C2H(CN)]2}, [MnTPP]{Ni[S2C2(CN)2]2}, and [MnTP'P]{Ni[S2C2(CN)2]2}, respectively, based upon the temperature at which maximum in the 10 Hz chi'(T) data occurs. [MnTP'P]{Ni[S2C2(CN)2]2} has a coercivity of 17,700 Oe and remanent magnetizations of 7250 emu Oe/mol at 2 K and 17 Oe and 850 emu Oe/mol at 5 K; hence, upon cooling it goes from being a soft magnet to being a very hard magnet. PMID- 16212380 TI - Counteranion-controlled water cluster recognition in a protonated octaamino cryptand. AB - Structural aspects of binding of water cluster and halides in the octaamino cryptand L (1,4,11,14,17,24,29,36 octaazapentacyclo[12.12.12.2.(6,9)2.(19,22)2(31,34)]tetratetraconta 6(43),7,9(44),19(41),20,22(42),31(39),32,34(40)-nonaene, N(CH2CH2NHCH2-p-xylyl CH2NHCH2CH2)3N) in a protonated state were examined. Crystallographic results show binding of the acyclic quasiplanar water tetramer [H4L(H2O)4](I)4.2.57H2O (1) in a tetraprotonated cryptand L having an iodide counteranion, where two water molecules reside inside the two tren-based cavity, bridged by a third water molecule, and a fourth external water molecule is hydrogen bonded to the bridged water molecule. In the case of complexes [H6L(Br)][(Br)6H].4H2O.2HBr (2) and [H6L(Cl)][(Cl)6H].10.86H2O (3), a single bromide and chloride occupied, respectively, the inside of the cryptand cavity, where L is in a hexaprotonated state. Monotopic recognition of bromide/chloride was observed at the center of the cryptand cavity where halides show C-H...halide interactions instead of the N H...halide interactions reported in the ditopic complexes of halides with the same cryptand, 5 and 6. Thermal analyses on 1-3 were carried out, and the data obtained distinctly differentiate water cluster complex 1 from the anion encapsulated cryptates 2 and 3. This study represents the first example of anion controlled cluster formation inside the cavity of a cryptand. PMID- 16212381 TI - Photon-, electron-, and proton-induced isomerization behavior of ferrocenylazobenzenes. AB - 3-, 4-, and 2-ferrocenylazobenzenes, 1, 2, and 3, respectively, and several derivatives of 1 were synthesized, and their photoisomerization behaviors were examined. The molecular structures of 1 and its derivatives, 2-chloro-5 ferrocenylazobenzene (5) and 3-ferrocenyl-4'-hydroxylazobenzene (11), were determined by X-ray diffraction analysis. 3-Ferrocenyl compound 1 undergoes reversible trans-to-cis isomerization with a single green light source and the Fe(III)/Fe(II) redox change. 4- and 2-Ferrocenyl compounds, 2 and 3, also respond to green light in addition to UV light, exciting the pi-pi* transition, but the cis molar ratio in the photostationary state (PSS) is lower than that of 1. The response to green light in 2 and 3 is caused by the MLCT (from Fe d orbital to azo pi* orbital) band excitation, while the character of the MLCT band, as estimated by time-dependent density functional theory calculations, differs between 1 and 2. The oxidized form of 2 undergoes facile cis-to-trans thermal isomerization. Both 1 and 2 undergo facile protonation and show proton-catalyzed cis-to-trans isomerization. Among the derivatives of 1, 2-chloro-5 ferrocenylazobenzene (5) exhibits the highest cis molar ratio (47%) in the PSS of green light irradiation. PMID- 16212382 TI - Phosphate triester hydrolysis promoted by an N2S(thiolate)Zn complex: mechanistic implications for the metal-dependent reactivity of peptide deformylase. AB - The zinc(II) complex (PATH)ZnOH, where PATH is an N2S(thiolate) ligand, has been investigated for its ability to promote the hydrolysis of the phosphate triester tris(4-nitrophenyl) phosphate (TNP). The hydrolysis of TNP was examined as a function of PATH-zinc(II) complex concentration, substrate concentration, and pH in a water/ethanol mixture (66:33 v/v) at 25 degrees C. The reaction is first order in both zinc(II) complex and substrate, and the second-order rate constants were derived from linear plots of the observed pseudo-first-order rate constants versus zinc complex concentration at different pH values. A pH-rate profile yielded a kinetic pK(a) of 8.52(5) for the zinc-bound water molecule and a pH independent rate constant of 16.1(7) M(-1) s(-1). Temperature-dependent studies showed linear Eyring behavior, yielding the activation parameters DeltaH++ = 36.9(1) kJ mol(-1) and DeltaS++ = -106.7(4) J mol(-1) K(-1). Interpretation of the kinetic data leads to the conclusion that hydrolysis of TNP takes place through a hybrid mechanism, in which the metal center plays a dual role of providing a nucleophilic hydroxide and activating the substrate through a Lewis acid effect. The synthesis and structural characterization of the related nickel(II) and iron(II) complexes [(PATH)2Ni2]Br2 (2) and (PATH)2Fe2Cl2 (3) are also described. Taken together, these data suggest a possible explanation for the low reactivity of the zinc(II) form of peptide deformylase as compared to the iron(II) form. PMID- 16212384 TI - Chelate ring size variations and their effects on coordination chemistry and catechol dioxygenase reactivity of iron(III) complexes. AB - The catechol dioxygenase reactivity of iron(III) complexes using tripodal ligands was investigated. Increasing, as well as decreasing, chelate ring sizes in the highly active complex [Fe(tmpa)(dbc)]B(C6H5)4 (tmpa = tris[(2 pyridyl)methyl]amine; dbc = 3,5-di-tert-butylcatecholate dianion), using related ligands, only resulted in decreased reactivity of the investigated compounds. A detailed low-temperature stopped-flow investigation of the reaction of dioxygen with [Fe(tmpa)(dbc)]B(C6H5)4 was performed, and activation parameters of DeltaH++ = 23 +/- 1 kJ mol(-1) and DeltaS++ = -199 +/- 4 J mol(-1) K(-1) were obtained. Crystal structures of bromo-(tetrachlorocatecholato-O,O')(bis((2-pyridyl)methyl) 2-pyridylamine-N,N',N'')-iron(III), (mu-oxo)-bis(bromo)(bis((2-pyridyl)methyl)-2 pyridylamine-N,N',N' ',N''')-diiron(III), dichloro-((2-(2-pyridyl)ethyl)bis((2 pyridyl)methyl)amine-N,N',N' ',N''')-iron(III) and (tetrachlorocatecholato O,O')((2-(2-pyridyl)ethyl)bis((2-pyridyl)methyl)amine-N,N',N' ',N''')-iron(III) are reported. PMID- 16212383 TI - Sulfate complexation of trivalent lanthanides probed by nanoelectrospray mass spectrometry and time-resolved laser-induced luminescence. AB - Sulfate complexation of lanthanides is of great interest for predicting speciation of radionuclides in natural environments. The formation of LaSO4+(aq) in HNO3/H2SO4 aqueous solutions of low ionic strength (I) was studied by nanoelectrospray ionization mass spectrometry (nanoESI-MS). Several gaseous species containing LaSO4+ were detected. The formation constant of LaSO4+(aq) was determined and extrapolated to I = 0 (log = 3.5 +/- 0.3) by using a simple specific ion interaction theory (SIT) formula. This value supports the potential of nanoESI-MS for the study of kinetically labile species. The species La(SO4)(2 ) was also detected. In addition, time-resolved laser-induced luminescence (TRLIL) was used to study Eu(III) speciation under ionic conditions of 0.02-0.05 M H+ (H2SO4/HClO4) and 0.4-2.0 M Na+ (Na2SO4/NaClO4). The data were interpreted with the species EuSO4+ (log = 3.7(8) +/- 0.1) and Eu(SO4)(2-) (log = 1.5 +/- 0.2). For extrapolating to I = 0, all of the major ions were taken into account through several SIT ion-pair parameters, epsilon. Most of the epsilon values were estimated by analogy to known parameters for similar ion-pair interactions using linear correlations, while epsilon(Eu)3+,SO4(2-) = 0.8(6) +/- 0.5 was fitted to the experimental data because, to date, SIT coefficients between multicharged species are not reported. The formation constants obtained here confirm some of those previously measured for Ln(III) and An(III) by various experimental techniques, and conversely do not give credit to the idea that in equilibrium conditions TRLIL and other spectroscopic techniques would provide stability constants of only inner-sphere complexes. The fluorescence lifetimes measured for EuSO4+ and Eu(SO4)(2-) were consistent with the replacement of one H2O molecule in the first coordination sphere of Eu3+ for each added SO4(2-) ligand, suggesting a monodentate SO4(2-) coordination. PMID- 16212385 TI - Vibrational spectra and gas-phase structure of N-methyl-S,S bis(trifluoromethyl)sulfimide, CH3N=S(CF3)2. AB - The molecular structure of N-methyl-S,S-bis(trifluoromethyl)sulfimide, CH3N=S(CF3)2, was determined by gas electron diffraction and quantum chemical calculations [B3LYP and MP2 with 6-31+G(2df,p) basis sets]. Furthermore, vibrational spectra, IR (gas) and Raman (liquid), were recorded. These spectra were assigned by comparison with analogous molecules and with calculated frequencies and intensities (HF, B3LYP, and MP2 with 6-311G basis sets). All experimental data and computational methods result in a single conformer with syn orientation of the CH3 group relative to the bisector of the two CF3 groups. The molecule possesses C1 symmetry, slightly distorted from CS symmetry. The N=S bond length in this compound [1.522(10) A] is longer than that in imidosulfur difluorides RN=SF2 [1.476(4) A - 1.487(5) A]. PMID- 16212386 TI - A deprotonated intermediate in the amide methanolysis reaction of an N4O-ligated mononuclear zinc complex. AB - Treatment of [(ppbpa)Zn](ClO4)2 (1(ClO4)2, ppbpa = N-((6-(pivaloylamido)-2 pyridyl)methyl)-N,N-bis((2-pyridyl)methyl)amine) with 1 equiv of Me(4)NOH.5H(2)O in methanol-acetonitrile solution results within minutes in the stoichiometric formation of a complex having a deprotonated amide, [(ppbpa-)Zn]ClO4 (3). Complex 3 has been characterized by 1H and 13C NMR, FTIR, and elemental analysis. Notably, upfield shifts of specific 1H NMR resonances of the amide-appended pyridyl moiety in 3, versus those found for 1(ClO4)2, indicate delocalization of the anionic charge within the amide-appended pyridyl donor of this complex. Heating of analytically pure 3 in methanol-acetonitrile results in amide alcoholysis. Overall, this alcoholysis reaction is second-order, with a first order dependence on both 3 and methanol. Analysis of the rate of decay of 3 as a function of temperature yielded activation parameters consistent with an intramolecular amide cleavage process (DeltaH++ = 15.0(3) kcal/mol, DeltaS++ = 33(1) eu). A possible reaction mechanism for amide alcoholysis is presented which involves reaction of the deprotonated amide intermediate 3 with methanol to produce a Lewis activated-type structure from which amide cleavage may be initiated. Additional support for this mechanistic pathway has been obtained through examination of the analogous ethanolysis reaction and via evaluation of the effect of varying steric hindrance near the amide carbonyl unit. PMID- 16212387 TI - The synthesis, structural, and spectroscopic characterization of uranium(IV) perrhenate complexes. AB - We report the synthesis, structural, and spectroscopic characterization of a series of uranium(IV)-perrhenato complexes. Three isostructural complexes with general formula [U(ReO4)4(L)4] (where L = tri-n-butylphosphine oxide/TBPO (2), triethyl phosphate/TEP (3), or tri-iso-butyl phosphate/TiBP (4)), have been synthesized, both through the photoreduction of ethanolic {UO2}2+ solutions and also via a novel U(IV) starting material, U(ReO4)4.5H2O (1). Compound 1 has also been used in the preparation of [U(ReO4)4(TPPO)3(CH3CN)].2CH3CN (5) and [U(ReO4)(DPPMO2)3(OH)][ReO4]2.2CH3CN (6), where TPPO represents triphenylphosphine oxide and DPPMO2 represents bis(diphenylphosphino)methane dioxide. All six complexes have been spectroscopically characterized using NMR, UV-vis-NIR, and IR techniques, with 2, 3, 5, and 6 also fully structurally characterized. The U atoms in compounds 2-6 all exhibit eight-coordinate geometry with up to four perrhenate groups in addition to three (DPPMO2 and TPPO) or four (TEP, TiBP, TBPO) coordinated organic ligands. In the case of compounds 5 and 6, the coordination of eight ligands to the U(IV) center is completed by the binding of a solvent molecule (CH3CN) and OH-, respectively. Solid-state physical analysis (elemental and thermogravimetric) and infrared spectroscopy are in agreement with the structural studies. The crystallographic data suggest that the strength of the U(IV)-O-donor ligand bonds decreases across the series R3PO > [ReO4]- > (RO)3PO. Solution-state IR and 31P NMR spectroscopy appear to be in agreement with these solid-state results. PMID- 16212388 TI - NMR, PGSE diffusion, and X-ray diffraction studies of lithium and potassium salts derived from diphenylphosphino(o-cyanophenyl)aniline and their crown ether complexes. AB - 1H, 31P, and 7Li pulsed-gradient spin-echo (PGSE) diffusion and variable temperature NMR results for THF solutions of the lithium and potassium salts derived from diphenylphosphino(o-cyanophenyl)aniline are reported and compared to the solid-state results obtained via X-ray diffraction studies. The solution results favor mononuclear salts, sometimes strongly ion paired, whereas the solid state data reveal dinuclear species. The structures of the products from reactions of these salts with crown ethers are determined via PGSE and 1H Overhauser NMR methods. PMID- 16212389 TI - Effect of pressure on proton-coupled electron transfer reactions of seven coordinate iron complexes in aqueous solutions. AB - For the first time, the effect of pressure on proton-coupled electron-transfer reactions of two selected seven-coordinate FeIII/II(H2L)(H2O)2 systems [where H2L = 2,6-diacetylpyridine-bis(semicarbazone) and 2,6-diacetylpyridine bis(semioxamazide), respectively] was examined. The acid-base equilibria of the different Fe(III/II) systems were investigated by spectrophotometric, potentiometric, and electrochemical titrations. On the basis of the obtained species distributions, the pH intervals in which the different protonated forms of the two studied systems exist were defined. In different pH ranges, a different number of protons (from 0 to 3 protons per electron) can be transferred during the redox process, which affects the change in the overall charge on the complexes. For all the different protonation forms of the studied complexes, the change in the redox potentials with pressure was measured and the redox reaction volume was obtained by high-pressure cyclic voltammetry. The results show that in the case of proton-coupled electron transfer, the reaction volume for the neutralization of protons contributes to the overall reaction volume. A linear correlation between Deltaz2 (change in the square of the charge) and the overall reaction volume of the complexes upon reduction, DeltaVcomplex0, was found. The average value of the intrinsic volume change for the selected seven-coordinate iron complexes was estimated from the intercept of the plot of DeltaVcomplex0 versus Deltaz2 to be 9.2 +/- 0.7 cm3 mol(-1). For the combined redox and protonation processes, the data are discussed in terms of linear correlations between Deltaz2 and the redox and neutralization reaction volumes reported in the literature. PMID- 16212390 TI - Infrared spectra of CH3-CrH, CH3-WH, CH2=WH2, and CH[triple bond]WH3 formed by activation of CH4 with Cr and W atoms. AB - Laser-ablated W atoms react with CH4 in excess argon to form the CH3-WH, CH2=WH2, and CH[triple bond]WH3 molecules with increasing yield in this order of product stability. These molecules are identified from matrix infrared spectra by isotopic substitution. Tungsten methylidene and methylidyne hydride molecules are reversibly interconverted by alpha-H transfers upon visible and ultraviolet irradiations. Matrix infrared spectra and DFT/B3LYP calculations show that CH[triple bond]WH3 is a stable molecule with C3v symmetry, but other levels of theory were required to describe agostic distortion for CH2=WH2. Analogous reactions with Cr gave only CH3-CrH, which is calculated to be by far the most stable product. PMID- 16212391 TI - Exploring the universality of unusual conformations of the 17-membered Pt(d(G*pG*)) macrochelate ring. Dependence of conformer formation on a change in bidentate carrier ligand from an sp3 to an sp2 nitrogen donor. AB - Early studies on cis-PtA2(d(G*pG*)) (A2 = diamine or two amines, G = N7 platinated G) and cis-Pt(NH3)2(d(G*pG*)) models for the key cisplatin-DNA cross link suggested that they exist exclusively or mainly as the HH1 conformer (HH1 = head-to-head G bases, with 1 denoting the normal direction of backbone propagation). These dynamic models are difficult to characterize. Employing carrier A2 ligands designed to slow dynamic interchange of conformers, we found two new conformers, DeltaHT (head-to-tail G* bases with a Delta chirality) and HH2 (with 2 denoting the backbone propagation direction opposite to normal). However, establishing that the non-HH1 conformations exist as an intrinsic feature of the 17-membered Pt(d(G*pG*)) ring requires exploring a range of different carrier ligands. Here we employ the planar aromatic sp(2) N-donor 5,5' Me(2)bipy (5,5'-dimethyl-2,2'-bipyridine) ligand, having a shape very different from those of previously used nonplanar sp(3) N-donor bidentate carrier ligands, which often bear NH groups. The 5,5'-Me(2)bipy H6 and H6' protons project toward the d(G*pG*) moiety and hinder the dynamic motion of 5,5'-Me(2)bipyPt(d(G*pG*)). We again found HH1, HH2, and DeltaHT conformers with typical properties, supporting the conclusions that the new DeltaHT and HH2 conformers exist universally in dynamic cis-PtA2(d(G*pG*)) adducts, including cis Pt(NH3)2(d(G*pG*)), and that the carrier ligand typically has little influence on the overall structure of the Pt(d(G*pG*)) macrocyclic ring of a given conformer. The sizes of the G H8 to H6/H6' NOE cross-peaks indicate little base canting in all 5,5'-Me(2)bipyPt(d(G*pG*)) conformers, suggesting that carrier-ligand NH groups favor the canting of one G base in the HH1 and HH2 conformers of typical cis-PtA2(d(G*pG*)) adducts. PMID- 16212392 TI - An electrochiroptical molecular switch: mechanistic and kinetic studies. AB - Previously, we reported the synthesis and preliminary characterization of Cu(I/II) complexes of N,N-bis(2-quinilylmethyl)-(l)-methionine (Zahn, S.; Canary, J. W. Science 2000, 288, 1404-7). The chemically oxidized and reduced forms of the complexes gave nearly mirror image circular dichroism (CD) spectra as a result of reorganization of the inner coordination sphere of the copper atom. The reorganization involved exchange of oxygen for sulfur in proceeding from the Cu(II) to Cu(I) oxidation state and corresponding ligand conformational changes required to accommodate this exchange. In this paper, we demonstrate that the complex can be triggered by electrochemical means. The electrochemical and stereochemical details of the redox-induced ligand reorganization were probed by independent synthesis of alternative chemical intermediates, CD spectroelectrochemistry, curve fitting of cyclic voltammograms, CD titration, and scanning electrochemical microscopy (SECM). A square-type mechanism was most consistent with the data: the Cu(II) complex is reduced followed by a ligand reorganization step to give the stable reduced form of the complex. The Cu(I) complex is then oxidized in a fast step followed by another ligand reorganization. A millisecond time scale rate was found by SECM for one of the key conformational conversion steps. PMID- 16212393 TI - General synthesis of di-mu-oxo dimanganese complexes as functional models for the oxygen evolving complex of photosystem II. AB - A series of complexes with the formula [Mn(III/IV)2(mu-O)2(L)2(X)2]3+ have been prepared in situ from Mn(II)LCl2 precursors by a general preparative method (L = terpy, Cl-terpy, Br-terpy, Ph-terpy, tolyl-terpy, mesityl-terpy, t Bu3-terpy, EtO terpy, py-phen, dpya, Me2N-terpy, or HO-terpy, and X = a labile ligand such as water, chloride, or sulfate). The parent complex, where L = terpy and X = water, is a functional model for the oxygen-evolving complex of photosystem II (Limburg, et al. J. Am. Chem. Soc. 2001, 123, 423-430). Crystals of Mn(II)(dpya)Cl2, Mn(II)(Ph-terpy)Cl2, Mn(II)(mesityl-terpy)Cl2, and an organic-soluble di-mu-oxo di-aqua dimanganese complex, [Mn(III/)(IV)2(mu-O)2(mesityl-terpy)2(OH2)2](NO3)3, were obtained and characterized by X-ray crystallography. Solutions of the in situ-formed di-mu-oxo dimanganese complexes were characterized by electrospray mass spectrometry, EPR spectroscopy, and UV-visible spectroscopy, and the rates of catalytic oxygen-evolving activity were assayed. The use of Mn(II)LCl2 precursors leads to higher product purity of the Mn dimers while achieving the 1:1 ligand to Mn stoichiometry appropriate for catalytic activity assay. These methods can be used to screen the catalytic activity of other di-mu-oxo dimanganese complexes generated by using a ligand library. PMID- 16212394 TI - Iron(III) coordination chemistry of alterobactin A: a siderophore from the marine bacterium Alteromonas luteoviolacea. AB - Alterobactin A is a siderophore produced by the oceanic bacterium Alteromonas luteoviolacea. The thermodynamic stability constant of the ferric alterobactin A (Alt-A) complex was estimated from electrochemical measurements on the basis of a previously reported linear relationship between the reduction potentials and the pH-independent stability constants for known iron(III) complexes. The reduction potential of the ferric alterobactin A complex determined by square wave voltammetry is -0.972 V vs SCE and reversible, corresponding to a thermodynamic stability constant of 10(51+/-2). Potentiometric titration of Fe(III)-Alt-A shows the release of six protons on complexation of Fe(III) to Alt-A. The 1H NMR resonances of the Ga(III)-Alt-A complex show that the C-4, C-5, and C-6 catecholate protons and the C(alpha) and C(beta) protons of both beta hydroxyaspartate moieties are shifted downfield relative to the free ligand, which along with the potentiometric titration data is consistent with a complex in which Fe(III) is coordinated by both catecholate oxygen atoms and both oxygen atoms of each beta-hydroxyaspartate. The UV-vis spectrum of Fe(III)-Alt-A is invariant over the pH range 4-9, indicating the coordination does not change over a wide pH range. In addition, in the absence of a coordinated metal ion, the serine ester of Alt-A hydrolyzes forming Alt-B. PMID- 16212395 TI - [Ru(phen)2(PHEHAT)]2+ and [Ru(phen)2(HATPHE)]2+: two ruthenium(II) complexes with the same ligands but different photophysics and spectroelectrochemistry. AB - The properties of two mononuclear Ru(II) complexes formed with the extended planar ligand PHEHAT depend drastically on the chelation site by the metallic ion. When the chelation takes place on the HAT site of the ligand (case of the novel complex [Ru(phen)2(HATPHE)]2+), the emission behavior is quite similar to that of [Ru(phen)2(HAT)]2+. In contrast, when the chelation is on the phen motif of the ligand (case of [Ru(phen)2(PHEHAT)]2+), the spectroscopic (absorption and emission) and electrochemical data for the complex do not obey the linear spectroelectrochemical correlation and the emission behavior is comparable to that of the extensively studied dppz complex ([Ru(phen/bpy)2(dppz)]2+). Thus, for [Ru(phen)2(PHEHAT)]2+, the emission lifetimes and intensities as a function of temperature exhibit a maximum for nitrile solvents. However, in contrast to the dppz case, at least three different states (two emitting and one dark) participate in the deactivation with different contributions depending on the temperature. These different contributions explain the observed maximum. Moreover, the fact that the solvent is liquid or frozen also influences the nature of the luminescent species. PMID- 16212398 TI - Selection of biomarkers by a multivariate statistical processing of composite metabonomic data sets using multiple factor analysis. AB - We introduce a statistical approach for integrating data from several analytical platforms. We illustrate this approach using (1)H-(13)C Heteronuclear Multiple Bond Connectivity nuclear magnetic resonance spectroscopy ((1)H-(13)C HMBC NMR) and Pyrolysis Metastable Atom Bombardment Time-of-Flight mass spectrometry (Py MAB-TOF-MS) to perform metabolic fingerprinting on cattle treated with anabolic steroids. Multiple factor analysis (MFA) integrates complementary aspects from NMR and MS data into a unique metabolic signature describing the biomarkers related to the dose-response. This work also indicates that, from a practical point of view, metabonomics and other "-omics" biotechnologies can benefit significantly from a generalized multi-platform integrative approach using multiple factor analysis. PMID- 16212399 TI - Homeostatic signature of anabolic steroids in cattle using 1H-13C HMBC NMR metabonomics. AB - We used metabonomics to discriminate the urinary signature of different anabolic steroid treatments in cattle having different physiological backgrounds (age, sex, and race). (1)H-(13)C heteronuclear multiple bonding connectivity NMR spectroscopy and multivariate statistical methods reveal that metabolites such as trimethylamine-N-oxide, dimethylamine, hippurate, creatine, creatinine, and citrate characterize the biological fingerprint of anabolic treatment. These urinary biomarkers suggest an overall homeostatic adaptation in nitrogen and energy metabolism. From results obtained in this study, it is now possible to consider metabonomics as a complementary method usable to improve doping control strategies to detect fraudulent anabolic treatment in cattle since the oriented global metabolic response provides helpful discrimination. PMID- 16212400 TI - Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues. AB - Renal cell carcinoma (RCC) tissue is composed of a mixture of neoplastic and normal cells, which complicate proteome analysis. The aim of our study was to investigate whether it is feasible to establish primary cell cultures of RCC and of renal cortex maintaining the tissue phenotype along with a more homogeneous and enriched cytological material. Fourteen (82.3%) primary cultures from 17 surgical cases were established and characterized by morphology, growth rate, immunocytochemistry, and molecular analysis performed by Real-time PCR, Western blotting, two-dimensional electrophoresis (2-DE), and mass spectrometry. Cultures showed >90% cytokeratine-positive epithelial cells. In primary tumor cultures, the molecular phenotype of manganese superoxide dismutase and heat shock protein 27 was the same as that found in tumor tissues with overexpression and increased number of isoforms. Moreover, 27 out 28 specific proteins and their isoforms, present in spots excised from 2-DE gel of cortex or RCC cultures, corresponded to those identified on the 2-DE tissue cortex reference map, suggesting that these primary cultures retain the proteomic profile of the corresponding tissues. PMID- 16212401 TI - Automated prescan function for scanning fluorescently stained 2D-PAGE gels. AB - To automate the acquisition of images from fluorescently stained gels, the power of the excitation laser(s) must be optimized for each sample to prevent spot saturation (or to allow unimportant spots to saturate) yet still retaining sensitivity. In this work, we describe the implementation and effectiveness of a pre-scan function in a robotic solution for the automation of 2D gel scanning. PMID- 16212402 TI - The platelet microparticle proteome. AB - Platelet-derived microparticles are the most abundant type of microparticle in human blood and contribute to many biologically significant processes. Here, we report the first proteomic analysis of microparticles generated from activated platelets. Using 1D SDS-PAGE and liquid chromatography coupled to a linear ion trap mass spectrometer, the identification of 578 proteins was accomplished using a minimum of 5 MS/MS detections of at least two different peptides for each protein. These microparticles displayed many proteins intrinsic to and well characterized on platelets. For example, microparticles in these experiments were found to contain membrane surface proteins including GPIIIa, GPIIb, and P selectin, as well other platelet proteins such as the chemokines CXCL4, CXCL7, and CCL5. In addition, approximately 380 of the proteins identified were not found in two previous studies of the platelet proteome. Since several of the proteins detected here have been previously implicated in microparticle formation and/or pathological function, it is hoped that this study will help fuel future work concerning the possible role of microparticles in various disease states. PMID- 16212403 TI - Reversed-phase high-performance liquid chromatographic prefractionation of immunodepleted human serum proteins to enhance mass spectrometry identification of lower-abundant proteins. AB - Serum analysis represents an extreme challenge due to the dynamic range of the proteins of interest, and the high structural complexity of the constituent proteins. In serum, the quantities of proteins and peptides of interest range from those considered "high abundance", present at 2-70% by mass of total protein, to those considered "low abundance", present at 10(-12) M or less. This range of analytical target molecules is outside the realm of available technologies for proteomic analysis. Therefore, in this study, we have developed a workflow toward addressing the complexity of these samples through the application of multidimensional separation techniques. The use of reversed-phase methods for the separation and fractionation of protein samples has been investigated, with the goal of developing an optimized serum separation for application to proteomic analysis. Samples of human serum were depleted of the six most abundant proteins, using an immunoaffinity LC method, then were separated under a variety of reversed-phase (RP) conditions using a macroporous silica C18 surface modified column material. To compare the qualities of the RP separations of this complex protein sample, absorbance chromatograms were compared, and fractions were collected for off-line SDS-PAGE and 2D-LC-MS/MS analysis. The column fractions were further investigated by determination of protein identities using either whole selected fractions, or gel bands excised from SDS-PAGE gels of the fractions. In either case samples underwent tryptic fragmentation and peptide analysis using MALDI-MS or LC-MS/MS. The preferred conditions for RP protein separation exhibited reproducibly high resolution and high protein recoveries (>98%, as determined by protein assay). Using the preferred conditions also permitted high column mass load, with up to 500 microg of protein well tolerated using a 4.6 mm ID x 50 mm column, or up to 1.5 mg on a 9.4 mm ID x 50 mm column. Elevated column temperature (80 degrees C) was observed to be a critical operational parameter, with poorer results observed at lower temperatures. The combination of sample simplification by immunoaffinity depletion combined with a robust and high recovery RP-HPLC fractionation yields samples permitting higher quality protein identifications by coupled LC-MS methods. PMID- 16212404 TI - Data-dependent electron capture dissociation FT-ICR mass spectrometry for proteomic analyses. AB - Electron capture dissociation (ECD) offers many benefits for the analysis of peptides and proteins, and consequently shows great potential for the field of proteomics. Recent developments have reduced the time scale required for ECD to milliseconds resulting in the technique's compatibility with on-line separation techniques, e.g., HPLC. Here, we demonstrate incorporation of ECD into a high throughput data-dependent LC-MS/MS approach for the analysis of proteomic samples. The approach is applied to analysis of the protein Fc-ROR2 isolated from chondrocytes and is the first example of LC-ECD-MS/MS of such a sample. Protein sequence coverage was 29%. Within that coverage, fifteen peptides were isolated and subjected to ECD. In most cases, the sequence tag generated by ECD was over 70% (in terms of the number of peptide backbone cleavages). The ECD data were searched against the nonredundant human NCBI database using the SEQUEST algorithm. Protein ROR2 was assigned, as was IgG (Fc domain). The results demonstrate the suitability of ECD as an integral technique in high-throughput proteomic strategies. PMID- 16212405 TI - Enrichment of phosphoproteins for proteomic analysis using immobilized Fe(III) affinity adsorption chromatography. AB - We described an efficient protocol to strongly enrich phosphoproteins from mixtures of total cellular proteins using homemade, recyclable Fe(III)-affinity columns. An integral feature of the method is the use of a detergent cocktail that allows use of different pHs for total protein extraction (pH 6.8) and for subsequent affinity capture of phosphoproteins (pH 3.4). Affinity captured proteins from rat fibroblasts were fractionated on 2D gels and random selection was identified by mass spectrometry. More than 85% of identified proteins were previously known to be phosphorylated. The specificity of the method was further validated by isolating proteins from (32)P labeled cells. Our comparison of the clusters of acidic residues in the captured proteins with acidic clusters in proteins of the rat genome indicates that affinity for phosphate groups dominates over adsorption of proteins with acidic clusters. PMID- 16212406 TI - Direct identification of ubiquitination sites on ubiquitin-conjugated CHIP using MALDI mass spectrometry. AB - The study of protein ubiquitination, a post-translational modification by ubiquitin, has emerged as one of the most active areas in biology because of the important role of this type of modification on the regulation of various cellular proteins. Advances in techniques for the determination and site mapping of protein ubiquitination can facilitate the elucidation of molecular mechanisms of this modification. We have recently described a novel method for identifying peptides containing ubiquitinated amino acid residues, based on the MALDI-MS/MS analysis of tryptic peptide derivatives. In particular, we have utilized N terminal sulfonation of these peptides to provide a unique fragmentation pattern that leads to the direct identification and sequencing of ubiquitin modified peptides. Here we present an application of this new method on the characterization of ubiquitin conjugated C-terminal Hsc70-interacting protein (CHIP), a recently identified U-box containing E3 enzyme. Three peptides bearing ubiquitination sites have been identified from the digest of ubiquitinated CHIP; one of these was a site on CHIP, while the other two were found on the ubiquitin molecules, demonstrating that sulfonation of tryptic peptides is a general and efficient method for characterizing protein ubiquitination. PMID- 16212407 TI - Investigation of the mouse serum proteome. AB - With the rapid assimilation of genomic information and the equally impressive developments in the field of proteomics, there is an unprecedented interest in biomarker discovery. Although human biofluids represent increasingly attractive samples from which new and more accurate disease biomarkers may be found, the intrinsic person-to-person variability in these samples complicates their discovery. One of the most extensively used animal models for studying human disease is mouse because, unlike humans, they represent a highly controllable experimental model system. Unfortunately, very little is known about the proteomic composition of mouse serum. In this study, a multidimensional fractionation approach on both the protein and the peptide level that does not require depletion of highly abundant serum proteins was combined with tandem mass spectrometry to characterize proteins within mouse serum. Over 12 300 unique peptides that originate from 4567 unique proteins-approximately 16% of all known mouse proteins-were identified. The results presented here represent the broadest proteome coverage in mouse serum and provide a foundation from which quantitative comparisons can be made in this important animal model. PMID- 16212408 TI - Ultra fast trypsin digestion of proteins by high intensity focused ultrasound. AB - Proteolytic digestion of proteins in seconds under an ultrasonic field provided by high-intensity focused ultrasound (HIFU) has been achieved. Successful in solution and in-gel tryptic digestion of proteins in 60 s or less was demonstrated by either MALDI-TOF mass spectrometry or liquid chromatography electrospray ion trap mass spectrometry (RP-HPLC-ESI-IT-MS/MS). The efficiency of this new procedure for protein digestion compared favorably with those attained using conventional overnight incubation methods. The performance of the method was also demonstrated by the specific identification of three proteins in a whole proteome in less than 1 h. The method greatly reduces the time needed for protein digestion, is of easy implementation, environmental friendly, and economic. Adaptation of this method to on-line procedures and robotic platforms could have promising applications in the proteomics field. PMID- 16212409 TI - Proteome approach to characterize the methylmalonate-semialdehyde dehydrogenase that is regulated by gibberellin. AB - Proteins regulated by gibberellin (GA) in rice were determined by proteome analysis. Proteins extracted from suspension culture cells of slr1, a constitutive GA response mutant of rice, were separated by two-dimensional polyacrylamide gel electrophoresis, and three proteins were greatly accumulated in the mutant. The most up-regulated protein was methylmalonate-semialdehyde dehydrogenase (MMSDH), and the amount of protein was 7-fold that of wild type. In this study, the function of MMSDH in rice was analyzed. MMSDH gene expression in suspension culture cells, roots, and leaf sheaths ofslr1 was higher than that in its wild-type. MMSDH expression in wild-type roots was increased by exogenous GA(3). Analyzed by in situ hybridization, MMSDH mRNA was expressed in root primordia of slr1, where cells are undergoing growth. MMSDH gene expression in the root zone of tissue differentiation was higher than in the elongation zone or meristem. Transgenic rice expressing antisense MMSDH showed that its seminal roots were thinner than that of control, and that the leaf sheath elongation was slightly inhibited compared to control. Concentrations of TCA cycle metabolites were decreased in the antisense plants as compared with the control plants, suggesting that acetyl-CoA was reduced in the antisense plants. These results suggest that one of the regulations by GA signal transduction including SLR1 is the expression of MMSDH, and that MMSDH may play a role in root development and leaf sheath elongation in rice. PMID- 16212410 TI - Proteomic evidence for roles for nucleolin and poly[ADP-ribosyl] transferase in drug resistance. AB - One-hundred twenty-four proteins have been identified in the soluble nuclear protein mixture from MCF-7 human breast cancer cells, of which more than 90% are classically categorized as nuclear proteins. Proteins were also studied from three drug resistant MDF-7 lines, selected previously from the same parent line by exposure to etoposide, to mitoxantrone, or to adriamycin in the presence of verapamil. Both quantitative gel comparisons and stable isotope labeling were used to identify a total of fourteen proteins whose abundances are altered by more than 2-fold in the three resistant lines. Several cytoskeleton proteins, cytokeratin 8, cytokeratin 19, septin 2, and alpha tropomyosin, are decreased in common across the three resistant cell lines. PARP-l (poly[ADP ribosyl]transefrase or connexion) is found to be less abundant in all three resistant lines. Nucleolin is more abundant in lines resistant to etoposide and mitoxantrone, while the mitotic checkpoint protein BUB 3 is more abundant in the line resistant to adriamycin/verapamil. PMID- 16212411 TI - Comprehensive phosphoproteome analysis in rice and identification of phosphoproteins responsive to different hormones/stresses. AB - Phosphoproteins in rice were detected by in vitro protein phosphorylation followed by two-dimensional polyacrylamide gel electrophoresis. Forty-four phosphoproteins were detected on a 2D-gel after in vitro protein phosphorylation of the crude extract from rice leaf sheath. Among the phosphoproteins detected, 42 were identified through analysis by Q-TOF MS/MS and/or MALDI-TOF MS. The largest percentage of the identified phosphoproteins are involved in signaling (30%), while 18% are involved in metabolism. When rice seedlings were treated with various hormones and stresses, it was observed that the phosphorylation of 13 proteins was enhanced differentially by different hormone and stress treatments. Furthermore, when the hormone/stress regulated phosphoproteins are compared in rice leaf sheath, leaf blade and root, only cytoplasmic malate dehydrogenase was found to be phosphorylated in all the tissues. Results suggest that in the phosphorylation cascade of rice, glycolytic metabolism processes and Ca(2+)-signaling seem to be important targets in response to hormones and stresses. Furthermore, the direct visualization of phosphoproteins by (32)P labeling and their mass spectrometric identification provides an accurate and reliable method of analyzing the rice phosphoproteome. PMID- 16212412 TI - Interresidue contacts in proteins and protein-protein interfaces and their use in characterizing the homodimeric interface. AB - The environment of amino acid residues in protein tertiary structures and three types of interfaces formed by protein-protein association--in complexes, homodimers, and crystal lattices of monomeric proteins--has been analyzed in terms of the propensity values of the 20 amino acid residues to be in contact with a given residue. On the basis of the similarity of the environment, twenty residues can be divided into nine classes, which may correspond to a set of reduced amino acid alphabet. There is no appreciable change in the environment in going from the tertiary structure to the interface, those participating in the crystal contacts showing the maximum deviation. Contacts between identical residues are very prominent in homodimers and crystal dimers and arise due to 2 fold related association of residues lining the axis of rotation. These two types of interfaces, representing specific and nonspecific associations, are characterized by the types of residues that partake in "self-contacts"--most notably Leu in the former and Glu in the latter. The relative preference of residues to be involved in "self-contacts" can be used to develop a scoring function to identify homodimeric proteins from crystal structures. Thirty-four percent of such residues are fully conserved among homologous proteins in the homodimer dataset, as opposed to only 20% in crystal dimers. Results point to Leu being the stickiest of all amino acid residues, hence its widespread use in motifs, such as leucine zippers. PMID- 16212413 TI - Uncovering the unfoldome: enriching cell extracts for unstructured proteins by acid treatment. AB - A method to enrich cell extracts in totally unfolded proteins was investigated. A literature search revealed that 14 of 29 proteins isolated by their failure to precipitate during perchloric acid (PCA) or trichloroacetic acid (TCA) treatment where also shown experimentally to be totally disordered. A near 100 000-fold reduction in yield was observed after 5% or 9% PCA treatment of total soluble E. coli protein. Despite this huge reduction, 158 and 142 spots were observed from the 5% and the 9% treated samples, respectively, on silver-stained 2-D SDS-PAGE gels loaded with 10 microg of protein. Treatment with 1% PCA was less selective with more visible spots and a greater than 3-fold higher yield. A substantial yield of unprecipitated protein was obtained after 3% TCA treatment, suggesting that the common use of TCA precipitation prior to 2-D gel analysis may result in loss of unstructured protein due to their failure to precipitate. Our preliminary analysis suggests that treating total protein extracts with 3-5% PCA and determining the identities of soluble proteins could be the starting point for uncovering unfoldomes (the complement of unstructured proteins in a given proteome). The 100 000-fold reduction in yield and concomitant reduction in number of proteins achieved by 5% PCA treatment produced a fraction suitable for analysis in its entirety using standard proteomic techniques. In this way, large numbers of totally unstructured proteins could be identified with minimal effort. PMID- 16212414 TI - Modeling biological variability in 2-D gel proteomic carcinogenesis experiments. AB - We propose a statistical method to model the underlying distribution of protein spot volumes in 2-D gels using a generalized model (GM). We apply this approach to discover mechanisms of chemical carcinogenesis in a rodent model. We generated 247 protein spots that were common to all gels (n = 18). Traditional statistical methods found 6.5% (13 out of 247) significant protein spots, our GM approach yielded a total of 53 (22.5%) differentially expressed protein spots. PMID- 16212415 TI - Phosphatase-directed phosphorylation-site determination: a synthesis of methods for the detection and identification of phosphopeptides. AB - Phosphopeptides can be difficult to detect and sequence by mass spectrometry (MS) due to low ionization efficiency and suppression effects in the MS mode, and insufficient fragmentation in the tandem MS (MS/MS) mode, respectively. To address this problem, we have developed a technique called Phosphatase-directed Phosphorylation-site Determination (PPD), which combines on-target phosphatase reactions, MALDI MS/MS of IMAC beads on target, and hypothesis-driven MS (HD-MS). In this method, on-target dephosphorylation experiments are conducted on IMAC bound phosphopeptides, because dephosphorylated peptides have, in general, higher MS sensitivities than the corresponding phosphopeptides. The detected dephosphorylated peptides are sequenced by MS/MS, which identifies the potentially phosphorylated peptide and the total number of Ser, Thr, or Tyr residues that could hypothetically be phosphorylated within that peptide. On the basis of this information, a mass list containing every possible phosphorylation state of each observed peptide (where 1 HPO(3) = 80 Da) is used to direct MALDI MS/MS on the phosphorylated peptides bound to IMAC beads at each theoretical mass from the list. If the peptide is present, the resulting MS/MS spectrum reveals the exact site(s) of phosphorylation in the peptide. We have demonstrated the applicability of PPD to the detection of in vivo phosphorylation sites on the Drosophila Stem Loop Binding Protein (dSLBP), and the complementarity of this new technique to conventional MS phosphorylation site mapping methods, since the phosphorylation sites in dSLBP could not be detected by other methods. PMID- 16212416 TI - Characterization of the Drosophila melanogaster mitochondrial proteome. AB - We have combined high-resolution two-dimensional (2-D) gel electrophoresis with mass spectrometry with the aim of identifying proteins represented in the 2-D gel database of Drosophila melanogaster mitochondria. First, we purified mitochondria from third instar Drosophila larvae and constructed a high-resolution 2-D gel database containing 231 silver-stained polypeptides. Next, we carried out preparative 2-D PAGE to isolate some of the polypeptides and characterize them by MALDI-TOF analysis. Using this strategy, we identified 66 mitochondrial spots in the database, and in each case confirmed their identity by MALDI-TOF/TOF analysis. In addition, we generated antibodies against two of the mitochondrial proteins as tools for characterizing the organelle. PMID- 16212417 TI - Biosensor-based micro-affinity purification for the proteomic analysis of protein complexes. AB - A biosensor-based micro-affinity purification method to recover protein binding partners and their complexes for down stream proteomics analysis has been developed using the BIAcore 3000 fitted with a prototype Surface Prep Unit (SPU). The recombinant GST-intracellular domain of E-cadherin or the recombinant GST beta-catenin binding domain of Adenomatous Polyposis Coli (APC) were immobilized onto the SPU and used to affinity purify binding partners from chromatographically enriched SW480 colon cancer cell lysates. A GST- immobilized surface was used as a control. Samples recovered from the SPU were subjected to SDS-PAGE with sensitive Coomassie staining followed by automated in-gel digestion and LC-MS/MS. The results obtained using the SPU were compared with similar experiments performed using Sepharose beads. PMID- 16212418 TI - Insights from modeling the 3D structure of DNA-CBF3b complex. AB - A 3-dimensional model for a DNA-protein interaction has been developed. The protein component is the 61-residue fragment (res. 11-71) of subunit b of the yeast centromeric DNA binding factor 3, CBF3b. The CBF3b fragments bind to the 17 base pairs (5'-CGGAGGACTGTCCTCCG-3') as a symmetric homodimer, with each folded into three distinct conformations: a compact, zinc-binding domain (res. 11-44); an extended linker (res. 45-57); and an alpha-helical dimerization element (res. 58-71). The DNA fragment in the complex is featured by a relatively straight conformation with only slight deviation from a standard B-structure, and a large part of the major groove not blocked by the protein. The large DNA open area provides the necessary space for the other subunits of CBF3 to bind coordinately with CBF3b, fully consistent with the observation that the cooperation of all four CBF3 components is absolutely required to constitute an activity that specifically interacts with centromere DNA. The model also provides a footing for further considering the possible binding arrangements of the other three subunits, namely CBF3a, CBF3c, and CBF3d. PMID- 16212419 TI - Phosphoproteome analysis of HeLa cells using stable isotope labeling with amino acids in cell culture (SILAC). AB - Identification of phosphorylated proteins remains a difficult task despite technological advances in protein purification methods and mass spectrometry. Here, we report identification of tyrosine-phosphorylated proteins by coupling stable isotope labeling with amino acids in cell culture (SILAC) to mass spectrometry. We labeled HeLa cells with stable isotopes of tyrosine, or, a combination of arginine and lysine to identify tyrosine phosphorylated proteins. This allowed identification of 118 proteins, of which only 45 proteins were previously described as tyrosine-phosphorylated proteins. A total of 42 in vivo tyrosine phosphorylation sites were mapped, including 34 novel ones. We validated the phosphorylation status of a subset of novel proteins including cytoskeleton associated protein 1, breast cancer anti-estrogen resistance 3, chromosome 3 open reading frame 6, WW binding protein 2, Nice-4 and RNA binding motif protein 4. Our strategy can be used to identify potential kinase substrates without prior knowledge of the signaling pathways and can also be applied to profiling to specific kinases in cells. Because of its sensitivity and general applicability, our approach will be useful for investigating signaling pathways in a global fashion and for using phosphoproteomics for functional annotation of genomes. PMID- 16212420 TI - Identification of protein fragments as pattern features in MALDI-MS analyses of serum. AB - The use of matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) to acquire spectral profiles has become a common approach to detect proteomic biomarkers of disease. MALDI-MS signals may represent both intact proteins as well as proteolysis products. Liquid chromatography-tandem mass spectrometry (LC MS/MS) analysis can tentatively identify the corresponding proteins Here, we describe the application of a data analysis utility called FragMint, which combines MALDI-MS spectral data with LC-MS/MS based protein identifications to generate candidate protein fragments consistent with both types of data. This approach was used to identify protein fragments corresponding to spectral signals in MALDI-MS analyses of unfractionated human serum. The serum also was analyzed by one-dimensional SDS-PAGE and bands corresponding to the MALDI-MS signal masses were excised and subjected to in-gel digestion and LC-MS/MS analysis. Database searches mapped all of the identified peptides to abundant blood proteins larger than the observed MALDI-MS signals. FragMint identified fragments of these proteins that contained the MS/MS identified sequences and were consistent with the observed MALDI-MS signals. This approach should be generally applicable to identify protein species corresponding to MALDI-MS signals. PMID- 16212421 TI - Coupling interaction between thromboxane A2 receptor and alpha-13 subunit of guanine nucleotide-binding protein. AB - G protein-coupled receptors (GPCRs) form a large superfamily of membrane proteins that play an essential role in modulating many vital physiological events, such as cell communication, neurotransmission, sensory perception, and chemotaxis. Understanding of the 3D (dimensional) structures of these receptors and their binding interactions with G proteins will help in the design of drugs for the treatment of GPCR-related diseases. By means of the approach of structural bioinformatics, the 3D structures of human alpha-13 subunit of guanine nucleotide binding protein (G alpha 13) and human thromboxane A2 (TXA2) receptor were developed. The former plays an important role in the control of cell growth that may serve as a prototypical G protein; the latter is a target for nitric oxide mediated desensitization that may serve as a prototypical GPCR. On the basis of the 3D models, their coupling interactions were investigated via docking studies. It has been found that the two proteins are coupled with each other mainly through the interaction between the minigene of G alpha 13 and the 3rd intracellular loop of the TXA2 receptor, consistent with the existing deduction in the literatures. However, it has also been observed via a close view that some residues of the TXA2 receptor that are sequentially far away but spatially quite close to the loop region are also involved in forming hydrogen bonds with the minigene of G alpha 13. These findings may provide useful information for conducting mutagenesis and reveal the molecular mechanism how the human TXA2 receptor interact with G alpha 13 to activate intracellular signaling. The findings may also provide useful insights for stimulating new therapeutic approaches by manipulating the interaction of the receptor with the relevant G proteins. PMID- 16212422 TI - Comparison of probability and likelihood models for peptide identification from tandem mass spectrometry data. AB - We evaluate statistical models used in two-hypothesis tests for identifying peptides from tandem mass spectrometry data. The null hypothesis H(0), that a peptide matches a spectrum by chance, requires information on the probability of by-chance matches between peptide fragments and peaks in the spectrum. Likewise, the alternate hypothesis H(A), that the spectrum is due to a particular peptide, requires probabilities that the peptide fragments would indeed be observed if it was the causative agent. We compare models for these probabilities by determining the identification rates produced by the models using an independent data set. The initial models use different probabilities depending on fragment ion type, but uniform probabilities for each ion type across all of the labile bonds along the backbone. More sophisticated models for probabilities under both H(A) and H(0) are introduced that do not assume uniform probabilities for each ion type. In addition, the performance of these models using a standard likelihood model is compared to an information theory approach derived from the likelihood model. Also, a simple but effective model for incorporating peak intensities is described. Finally, a support-vector machine is used to discriminate between correct and incorrect identifications based on multiple characteristics of the scoring functions. The results are shown to reduce the misidentification rate significantly when compared to a benchmark cross-correlation based approach. PMID- 16212423 TI - From dormant to germinating spores of Streptomyces coelicolor A3(2): new perspectives from the crp null mutant. AB - The complete understanding of the morphological differentiation of streptomycetes is an ambitious challenge as diverse sensors and pathways sensitive to various environmental stimuli control the process. Germination occupies a particular position in the life cycle as the good achievement of the process depends on events occurring both during the preceding sporulation and during germination per se. The cyclic AMP receptor protein (crp) null mutant of Streptomyces coelicolor, affected in both sporulation and germination, was therefore presented as a privileged candidate to highlight new proteins involved in the shift from dormant to germinating spores. Our multidisciplinary approach-combining in vivo data, the analysis of spores morphological properties, and a proteome study-has shown that Crp is a central regulatory protein of the life cycle in S. coelicolor; and has identified spores proteins with statistically significant increased or decreased expression that should be listed as priority targets for further investigations on proteins that trigger both ends of the life cycle. PMID- 16212424 TI - Simple and robust two-layer matrix/sample preparation method for MALDI MS/MS analysis of peptides. AB - Recent advances in MALDI MS/MS instrumentation allow a high degree of automation in the efficient detection of peptide fragment ions that can be used for protein identification. However, the performance of the technique is dependent on the MALDI sample preparation. We present a simple and robust two-layer sample preparation method tailored for sensitive and reproducible generation of MALDI MS/MS data. This method produces a strong and uniform crystal layer which allows acquisition of high quality MS/MS spectra over the entire sample surface area. Furthermore, due to its crystal strength, the matrix/sample layer can be washed extensively on target, enabling direct analysis of samples containing impurities, such as salts and surfactants. This method is demonstrated to be very useful in routine analysis of in-gel tryptic digests of silver-stained protein gel spots, without the need of desalting steps or hunting for "hot" spots. As an example, seven threonine-phosphorylated proteins involved in signal transduction in response to growth factor stimulation within the lipid raft fractions of the IMR5 neuroblastoma cells have been identified using differential gel display, in-gel digestion and MALDI MS/MS with the new two-layer sample preparation method. Some of these proteins have the functions of maintaining raft structure or cell signaling. PMID- 16212425 TI - Detection and identification of protein interactions of S100 proteins by ProteinChip technology. AB - The aim of this work was to establish an approach for identification of protein interactions. This assay used an anti-S100A8 antibody coupled on beads and incubated with cell extract. The bead eluates were analyzed using ProteinChip technology and subsequently subjected to an appropriate digestion. Molecular masses of digestion fragments were determined by SELDI-MS, and database analysis revealed S100A10 as interacting protein. This result was confirmed by co immunoprecipitation and immunocapturing. Using S100A10 as new bait, a specific interaction with S100A7 was detectable. PMID- 16212426 TI - Effect of immunoaffinity depletion of human serum during proteomic investigations. AB - Controversy exists regarding the proper mining of the human serum proteome. Because of the analytical challenges of accurately measuring samples containing a very large dynamic range of protein concentrations, current practices have employed depletion of the highly abundant housekeeping serum proteins, such as albumin and immunoglobins. There is question as to the selectivity of depletion, namely, is there loss of other non abundant serum proteins along with albumin, haptoglobin and other commonly depleted proteins. In this study, human serum was analyzed with and without immunoaffinity depletion of the six most abundant proteins by multidimensional liquid chromatography tandem mass spectrometry. Two replicates of each experiment were conducted and compared against one another. In both depleted and nondepleted replicates there was a 73% and 72% overlap of identified peptides and a 64% and 78% overlap of identified proteins, respectively. Of 262 unique proteins identified in the four experiments, 82 were found in common to all four experiments, 142 unique to the depleted serum, and 38 unique to the nondepleted serum. Although serum depletion of highly abundant proteins significantly increased the number of proteins identified, both the degree of sample complexity and this depletion method resulted in a nonselective loss of other proteins. PMID- 16212427 TI - Identification and analysis of multivalent proteolytically resistant peptides from gluten: implications for celiac sprue. AB - Dietary gluten proteins from wheat, rye, and barley are the primary triggers for the immuno-pathogenesis of Celiac Sprue, a widespread immune disease of the small intestine. Recent molecular and structural analyses of representative gluten proteins, most notably alpha- and gamma-gliadin proteins from wheat, have improved our understanding of these pathogenic mechanisms. In particular, based on the properties of a 33-mer peptide, generated from alpha-gliadin under physiological conditions, a link between digestive resistance and inflammatory character of gluten has been proposed. Here, we report three lines of investigation in support of this hypothesis. First, biochemical and immunological analysis of deletion mutants of alpha-2 gliadin confirmed that the DQ2 restricted T cell response to the alpha-2 gliadin are directed toward the epitopes clustered within the 33-mer. Second, proteolytic analysis of a representative gamma-gliadin led to the identification of another multivalent 26-mer peptide that was also resistant to further gastric, pancreatic and intestinal brush border degradation, and was a good substrate of human transglutaminase 2 (TG2). Analogous to the 33 mer, the synthetic 26-mer peptide displayed markedly enhanced T cell antigenicity compared to monovalent control peptides. Finally, in silico analysis of the gluten proteome led to the identification of at least 60 putative peptides that share the common characteristics of the 33-mer and the 26-mer peptides. Together, these results highlight the pivotal role of physiologically generated, proteolytically stable, TG2-reactive, multivalent peptides in the immune response to dietary gluten in Celiac Sprue patients. Prolyl endopeptidase treatment was shown to abolish the antigenicity of both the 33-mer and the 26-mer peptides, and was also predicted to have comparable effects on other proline-rich putatively immunotoxic peptides identified from other polypeptides within the gluten proteome. PMID- 16212428 TI - Characterization of proteins in human pancreatic cancer serum using differential gel electrophoresis and tandem mass spectrometry. AB - The purpose of this study was to develop techniques for identifying cancer biomarkers in human serum using differential in-gel electrophoresis (DIGE), and characterizing the protein biomarkers using tandem mass spectrometry (MS/MS). A major problem in profiling protein expression by DIGE comes from the presence of high concentrations of a small number of proteins. Therefore, serum samples were first chromatographed using an immunoaffinity HPLC column (Agilent Technologies), to selectively remove albumin, immunoglobulins, transferrin, haptoglobin, and antitrypsin. Serum samples from three individuals with pancreatic cancer and three individuals without cancer were compared. Serum samples were processed using the immunoaffinity column. Differential protein analysis was performed using DIGE. A total of 56 protein spot-features were found to be significantly increased and 43 significantly decreased in cancer serum samples. These spot features were excised, trypsin digested, and analyzed by MALDI/TOF/TOF (4700 Proteomics Analyzer, Applied Biosystems). We identified 24 unique proteins that were increased and 17 unique proteins that were decreased in cancer serum samples. Western blot analysis confirmed increased levels of several of these proteins in the pancreatic cancer serum samples. In an independent series of serum samples from 20 patients with pancreatic cancer and 14 controls, increased levels of apolipoprotein E, alpha-1-antichymotrypsin, and inter-alpha-trypsin inhibitor were found to be associated with pancreatic cancer. These results suggest that affinity column enrichment and 2-D DIGE can be used to identify numerous proteins differentially expressed in serum from individuals with pancreatic cancer. PMID- 16212429 TI - I-DIRT, a general method for distinguishing between specific and nonspecific protein interactions. AB - Isolation of protein complexes via affinity-tagged proteins provides a powerful tool for studying biological systems, but the technique is often compromised by co-enrichment of nonspecifically interacting proteins. We describe a new technique (I-DIRT) that distinguishes contaminants from bona fide interactors in immunopurifications, overcoming this most challenging problem in defining protein complexes. I-DIRT will be of broad value for studying protein complexes in biological systems that can be metabolically labeled. PMID- 16212430 TI - Human tissue profiling with multidimensional protein identification technology. AB - Profiling of tissues and cell types through systematic characterization of expressed genes or proteins shows promise as a basic research tool, and has potential applications in disease diagnosis and classification. We used multidimensional protein identification protein identification technology (MudPIT) to analyze proteomes for enriched nuclear extracts of eight human tissues: brain, heart, liver, lung, muscle, pancreas, spleen, and testis. We show that the method is approximately 80% reproducible. We address issues of relative abundance, tissue-specificity, and selectivity, and the significance of proteins whose expression does not correlate with that of the corresponding mRNA. Surprisingly, most proteins are detected in a single tissue. These proteins tend to fulfill specialist (and potentially tissue-specific) functions compared to proteins expressed in two or more tissues. PMID- 16212431 TI - AUDENS: a tool for automated peptide de novo sequencing. AB - We present AUDENS, a new platform-independent open source tool for automated de novo sequencing of peptides from MS/MS data. We implemented a dynamic programming algorithm and combined it with a flexible preprocessing module which is designed to distinguish between signal and other peaks. By applying a user-defined set of heuristics, AUDENS screens through the spectrum and assigns high relevance values to putative signal peaks. The algorithm constructs a sequence path through the MS/MS spectrum using the peak relevances to score each suggested sequence path, i.e., the corresponding amino acid sequence. At present, we consider AUDENS a prototype that unfolds its biggest potential if used in parallel with other de novo sequencing tools. AUDENS is available open source and can be downloaded with further documentation at http://www.ti.inf.ethz.ch/pw/software/audens/ . PMID- 16212432 TI - Characterization of vacuolar membrane proteins changed in rice root treated with gibberellin. AB - Rice vacuolar membrane proteins changed by gibberellin (GA) were analyzed using a proteome approach. Vacuolar membrane fractions were isolated using a discontinuous sucrose/sorbitol system and 10 proteins increased in vacuolar membrane of the root, treated with GA(3) as compared with control. Fructose-1,6 bisphosphate aldolase C-1 and vacuolar H(+)-ATPase (V-ATPase) increased in root vacuolar membrane by GA(3) interacted in rice roots. It suggests that aldolase C 1 regulates the V-ATPase mediated control of cell elongation that determines root growth. PMID- 16212433 TI - Dietary effects of copper and iron deficiency on rat intestine: a differential display proteome analysis. AB - Copper and iron are cofactors of many metallo-proteins that accomplish vital functions, such as oxygen and electron transport. Specific metabolic pathways have been selected through evolution, although still not fully elucidated, to confine the dangerous reactivity of their free ionic forms. Inadequate supply of both metals can severely affect basic physiological functions. A differential analysis of the rat intestinal proteome evidenced the following dietary copper- and iron-deficiencies, i.e., significant changes in the levels of proteins belonging to different functional classes (glucose and fatty acid metabolism, molecular chaperones, cytoskeleton plasticity, vitamin transporters). The presented results bring new perspectives to understand the role of copper and iron in the metabolic pathways and provide novel diagnostic tools to characterize the effects of subclinical deficiencies of both metals in unbalanced nutritional disorders. PMID- 16212434 TI - Identification and characterization of the Sulfolobus solfataricus P2 proteome. AB - Via combined separation approaches, a total of 1399 proteins were identified, representing 47% of the Sulfolobus solfataricus P2 theoretical proteome. This includes 1323 proteins from the soluble fraction, 44 from the insoluble fraction and 32 from the extra-cellular or secreted fraction. We used conventional 2 dimensional gel electrophoresis (2-DE) for the soluble fraction, and shotgun proteomics for all three cell fractions (soluble, insoluble, and secreted). Two gel-based fractionation methods were explored for shotgun proteomics, namely: (i) protein separation utilizing 1-dimensional gel electrophoresis (1-DE) followed by peptide fractionation by iso-electric focusing (IEF), and (ii) protein and peptide fractionation both employing IEF. Results indicate that a 1D-IEF fractionation workflow with three replicate mass spectrometric analyses gave the best overall result for soluble protein identification. A greater than 50% increment in protein identification was achieved with three injections using LC ESI-MS/MS. Protein and peptide fractionation efficiency; together with the filtration criteria are also discussed. PMID- 16212435 TI - Characterization of pepsinogen C as a potential biomarker for gastric cancer using a histo-proteomic approach. AB - We analyzed 74 cryostat sections of central gastric tumor, tumor margin, and normal gastric epithelium using ProteinChip Arrays and SELDI-TOF MS. One peak was significantly down-regulated in tumor tissue (P = 1.43 x 10(-6)) and identified as pepsinogen C using MS/MS analysis and immunodepletion. This signal was further characterized by immunohistochemistry. This work demonstrates that differentially expressed signals can be identified and assessed using a proteomic approach comprising tissue-microdissection, protein profiling, and immunohistochemistry. PMID- 16212436 TI - Chemically modified, immobilized trypsin reactor with improved digestion efficiency. AB - Tryptic digestion followed by identification using mass spectrometry is an important step in many proteomic studies. Here, we describe the preparation of immobilized, acetylated trypsin for enhanced digestion efficacy in integrated protein analysis platforms. Complete digestion of cytochrome c was obtained with two types of modified-trypsin beads with a contact time of only 4 s, while corresponding unmodified-trypsin beads gave only incomplete digestion. The digestion rate of myoglobin, a protein known to be rather resistant to proteolysis, was not altered by acetylating trypsin and required a buffer containing 35% acetonitrile to obtain complete digestion. The use of acetylated trypsin beads led to fewer interfering tryptic autolysis products, indicating an increased stability of this modified enzyme. Importantly, the modification did not affect trypsin's substrate specificity, as the peptide map of myoglobin was not altered upon acetylation of immobilized trypsin. Kinetic digestion experiments in solution with low-molecular-weight substrates and cytochrome c confirmed the increased catalytic efficiency (lower K(M) and higher k(cat)) and increased resistance to autolysis of trypsin upon acetylation. Enhancement of catalytic efficiency was correlated with the number of acetylations per molecule. The favorable properties of the new chemically modified trypsin reactor should make it a valuable tool in automated protein analysis systems. PMID- 16212437 TI - Comparative plasma proteome analysis of lymphoma-bearing SJL mice. AB - In SJL mice, growth of RcsX lymphoma cells induces an inflammatory response by stimulating V(beta)16+ T cells. During inflammation, various serum protein levels can increase (e.g., acute phase reactants) or decrease (e.g., albumin), and most of these altered proteins are thus potential biomarkers. Although blood plasma is a valuable and promising sample for biomarker discovery for diseases or for novel drug targets, its proteome is complex. To address this, we have focused on a comprehensive comparison of the plasma proteomes from normal and RcsX-tumor bearing SJL mice using the 1D-Gel-LC-MS/MS method after removing albumin and immunoglobulins. This analysis resulted in the identification of a total of 1079 nonredundant mouse plasma proteins; more than 480 in normal and 790 in RcsX-tumor bearing SJL mouse plasma. Of these, only 191 proteins were found in common. The molecular weights ranged from 2 to 876 kDa, covering the pI values between 4.22 and 12.09, and included proteins with predicted transmembrane domains. By comparing the plasma proteomic profile of normal and RcsX-tumor-bearing SJL mice, we found significant changes in the levels of many proteins in RcsX-tumor-bearing mouse plasma. Most of the up-regulated proteins were identified as acute-phase proteins (APPs). Also, several unique proteins i.e., haptoglobin, proteosome subunits, fetuin-B, 14-3-3 zeta, MAGE-B4 antigen, etc, were found only in the tumor-bearing mouse plasma; either secreted, shed by membrane vesicles, or externalized due to cell death. These results affirm the effectiveness of this approach for protein identification from small samples, and for comparative proteomics in potential animal models of human disorders. PMID- 16212438 TI - STEM: a software tool for large-scale proteomic data analyses. AB - We describe the software, STEM (STrategic Extractor for Mascot's results), which efficiently processes large-scale mass spectrometry-based proteomics data. V (View)-mode evaluates the Mascot peptide identification dataset, removes unreliable candidates and redundant assignments, and integrates the results with key information in the experiment. C (Comparison)-mode compares peptide coverage among multiple datasets and displays proteins commonly/specifically found therein, and processes data for quantitative studies that utilize conventional isotope tags or tags having a smaller mass difference. STEM significantly improves throughput of proteomics study. PMID- 16212439 TI - Methods for the detection of paxillin post-translational modifications and interacting proteins by mass spectrometry. AB - Methods for the simultaneous identification of interacting proteins and post translational modifications of the focal adhesion adapter protein, paxillin, are presented. The strategy includes (1) lower-level, transient transfection of FLAG GFP-Paxillin into HEK293 cells, (2) incubation of cells with phosphatase inhibitors prior to lysis, (3) purification of paxillin by anti-FLAG immunoprecipitation, (4) analysis of peptides generated from on-beads digestion using LTQ-FT or LTQ-ETD mass spectrometry, and (5) enrichment of phosphopeptide methyl esters with IMAC. Using the above strategies, we identify 29 phosphorylation sites (19 novel and 10 previously reported) and a novel glycosylation site on Ser 74. Furthermore, with this method, we simultaneously detect 10 co-purifying proteins which are present in focal adhesion complexes. Extension of these methods to other substrates should facilitate generation of global phosphorylation maps and protein-protein interactions for any protein of interest. PMID- 16212440 TI - Parallel tandem: a program for parallel processing of tandem mass spectra using PVM or MPI and X!Tandem. AB - A method for the rapid correlation of tandem mass spectra to a list of protein sequences in a database has been developed. The combination of the fast and accurate computational search algorithm, X!Tandem, and a Linux cluster parallel computing environment with PVM or MPI, significantly reduces the time required to perform the correlation of tandem mass spectra to protein sequences in a database. A file of tandem mass spectra is divided into a specified number of files, each containing an equal number of the spectra from the larger file. These files are then searched in parallel against a protein sequence database. The results of each parallel output file are collated into one file for viewing through a web interface. Thousands of spectra can be searched in an accurate, practical, and time effective manner. The source code for running Parallel Tandem utilizing either PVM or MPI on Linux operating system is available from http://www.thegpm.org. This source code is made available under Artistic License from the authors. PMID- 16212442 TI - Prediction of functional class of the SARS coronavirus proteins by a statistical learning method. AB - The complete genome of severe acute respiratory syndrome coronavirus (SARS-CoV) reveals the existence of putative proteins unique to SARS-CoV. Identification of their function facilitates a mechanistic understanding of SARS infection and drug development for its treatment. The sequence of the majority of these putative proteins has no significant similarity to those of known proteins, which complicates the task of using sequence analysis tools to probe their function. Support vector machines (SVM), useful for predicting the functional class of distantly related proteins, is employed to ascribe a possible functional class to SARS-CoV proteins. Testing results indicate that SVM is able to predict the functional class of 73% of the known SARS-CoV proteins with available sequences and 67% of 18 other novel viral proteins. A combination of the sequence comparison method BLAST and SVMProt can further improve the prediction accuracy of SMVProt such that the functional class of two additional SARS-CoV proteins is correctly predicted. Our study suggests that the SARS-CoV genome possibly contains a putative voltage-gated ion channel, structural proteins, a carbon oxygen lyase, oxidoreductases acting on the CH-OH group of donors, and an ATP binding cassette transporter. A web version of our software, SVMProt, is accessible at http://jing.cz3.nus.edu.sg/cgi-bin/svmprot.cgi . PMID- 16212441 TI - Francisella tularensis proteome: low levels of ASB-14 facilitate the visualization of membrane proteins in total protein extracts. AB - Proteomic analysis of bacterial pathogens isolated from in vivo sources, such as infected tissues, provides many challenges not the least of which is the limited quantity of sample available for analysis. It is, therefore, highly desirable to develop a one-step cellular lysis and protein solubilization method that minimizes protein losses and allows the maximum possible coverage of the proteome. Here, we have used standard sample buffer constituents including urea, thiourea and DTT, but varied the detergent composition of the buffers in order to achieve the best quality of gels and the greatest spot resolution. We found that the most efficient solubilizing solution in this case consisted of 7 M urea, 2 M thiourea, 1% DTT, 0.5% amidosulfobetaine-14 (ASB-14) and 4% 3-[(3 Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). Inclusion of low levels of ASB-14 in solutions allowed visualization of a subset of 24 new protein spots in the Live Vaccine Strain (LVS) of Francisella tularensis and 21 spots in a virulent A-strain of the pathogen. Further investigation showed that 15 of the 24 enriched LVS spots were membrane or membrane-associated proteins suggesting that the optimized lysis and solubilization solution aids in the detection of more hydrophobic proteins. This methodology is now being applied to the analysis of Francisella obtained from in vivo sources. PMID- 16212443 TI - Optimizing thiophosphorylation in the presence of competing phosphorylation with MALDI-TOF-MS detection. AB - Thiophosphorylation provides a metabolically stable, chemically reactive phosphorylation analogue for analyzing the phosphoproteome in vitro and in vivo. We developed a MALDI-TOF-MS based assay for optimizing thiophosphopeptide production by a kinase even in the presence of Mg(2+) and ATP. We found that Abl kinase thiophosphorylation rates can be "rescued" using Mn(2+) in the presence of Mg(2+). Under our ideal conditions, titration of Mn(2+) and ATPgammaS in the presence of Mg(2+) allowed relatively rapid, highly specific thiophosphorylation by Abl tyrosine kinase, both as purified enzyme and in complex cell extracts. PMID- 16212444 TI - Impact of replicate types on proteomic expression analysis. AB - In expression proteomics, the samples utilized within an experimental design may include technical, biological, or pooled replicates. This manuscript discusses various experimental designs and the conclusions that can be drawn from them. Specifically, it addresses the impact of mixing replicate types on the statistical analysis which can be performed. This study focuses on difference gel electrophoresis (DiGE), but the issues are equally applicable to all quantitative methodologies assessing relative changes in protein expression. PMID- 16212446 TI - Intensive behavioral treatment for children with autism: four-year outcome and predictors. AB - Twenty-four children with autism were randomly assigned to a clinic-directed group, replicating the parameters of the early intensive behavioral treatment developed at UCLA, or to a parent-directed group that received intensive hours but less supervision by equally well-trained supervisors. Outcome after 4 years of treatment, including cognitive, language, adaptive, social, and academic measures, was similar for both groups. After combining groups, we found that 48% of all children showed rapid learning, achieved average posttreatment scores, and at age 7, were succeeding in regular education classrooms. Treatment outcome was best predicted by pretreatment imitation, language, and social responsiveness. These results are consistent with those reported by Lovaas and colleagues (Lovaas, 1987; McEachin, Smith, & Lovaas, 1993). PMID- 16212447 TI - Longitudinal assessment of stereotypic, proto-injurious, and self-injurious behavior exhibited by young children with developmental delays. AB - Twelve children (CA, 12 to 32 months) with developmental delay were observed in their homes during monthly analogue functional analysis probes to document patterns of emerging self-injurious behavior. Two patterns of emerging self injury were observed for 5 participants: (a) The topography and functional analysis pattern remained the same, but the behavior eventually caused tissue damage; or (b) a new topography emerged that was similar to an established stereotypic motor behavior. Functional analysis results were inconclusive for the majority of target behaviors across participants due to undifferentiated responding across conditions. One participant exhibited two topographies that appeared to become sensitive to positive reinforcement over time. Results are discussed in terms of implications for future research on early intervention and prevention of self-injury. PMID- 16212448 TI - Life course impacts of mild intellectual deficits. AB - We identified 201 individuals who obtained IQs of 85 or below in high school and participated in the Wisconsin Longitudinal Study (a prospective longitudinal study that followed sample members from age 18 through age 53). Their life course development was contrasted with their siblings who obtained IQs above 100. Life course outcomes were assessed in five domains: Education and Occupational Attainment, Family Formation, Social Participation and Support, Physical Health, and Psychological Well-Being. Compared to their higher IQ sibling, low-IQ individuals completed less schooling, had less prestigious occupations, rated themselves less physically healthy, and reported lower levels of psychological well-being. Differences were not evident, however, in patterns of family formation, social support, job satisfaction, or objective measures of physical health. PMID- 16212449 TI - Psychometric evaluation of a self-report measure of depression for individuals with mental retardation. AB - The psychometric properties of the Self-Report Depression Questionnaire (SRDQ) were evaluated, extending a previous assessment of this instrument. Data from two independent studies (Esbensen, 2004; Seltzer & Krauss, 1989) were pooled to generate a sample of 192 individuals with primarily mild or moderate mental retardation. Reliability estimates of this questionnaire were good to excellent and corroborated prior findings. In addition, the measure was found to have sound validity as evidenced by convergent validity, discriminant validity, and predictive validity and by comparing individuals with and without clinical diagnoses of depression. Thus, the SRDQ appears to be a reliable and valid measure of depressive symptomatology for individuals with mild or moderate mental retardation. Screening procedures are suggested. PMID- 16212450 TI - Repetition priming in adults with Williams syndrome: age-related dissociation between implicit and explicit memory. AB - We examined implicit and explicit memory in adults with Williams syndrome. An age related dissociation was found; repetition priming (reflecting implicit memory) did not show change with age, but free recall (reflecting explicit memory) was markedly reduced. We also compared the performance of adults with Williams syndrome to adults with Down syndrome and those with unspecified mental retardation. A similar dissociation was observed in adults with Down syndrome but not in adults with unspecified mental retardation. An IQ-related dissociation was also found. Implicit and explicit memory, therefore, show different degrees of association with age and IQ, supporting theories of these memory processes. Results also suggest that Williams syndrome, similar to Down syndrome, may be associated with precocious aging, resulting in the loss of some cognitive abilities. PMID- 16212452 TI - Feasibility trial of carotid stenting with and without an embolus protection device. AB - PURPOSE: To evaluate the feasibility of stenting and cerebral protection in patients with carotid bifurcation disease. METHODS: Carotid stenting with self expanding nitinol stents was evaluated in 261 patients (157 men; mean age 70.9+/ 9.7 years, range 41-90) treated at 33 clinical sites. Among these patients, 141 (54.3%) were symptomatic, and 71 (27.2%) had postendarterectomy restenotic lesions. An embolus protection device became available in the latter third of the study. The primary endpoint of the study was the 30-day composite occurrence of major adverse clinical events (all deaths, strokes, or myocardial infarctions [MI]). At 1 year, the endpoint included all deaths, strokes, or MIs at 30 days, plus all deaths or ipsilateral strokes between 31 days and 1 year. RESULTS: SMART stents were placed in 147 (56%) patients and Precise stents in 107 (41%); 7 (3%) patients were treated with non-study stents. Lesion success, defined as <30% postprocedural residual stenosis, was achieved in 249 (95%). An AngioGuard filter was used in 85 patients; of 90 filters employed, 78 (87%) were successfully deployed and retrieved. Visible material was noted in 42 (54%) retrieved filters. One (0.4%) stent thrombosis occurred at <30 days, and no restenotic lesions >80% were noted in up to 1 year of follow-up. At 30 days, stroke occurred in 16 (6.1%) patients, MI in 1 (0.4%), and 2 (0.8%) patients died. The 30-day rate of stroke, MI, or death was 6.9%. Major ipsilateral stroke occurred in 4 (1.5%) patients between 0 and 30 days; no new major ipsilateral strokes were noted up to 1 year. By Kaplan-Meier analysis, the 1-year risk of stroke, MI, or death was 10.9%+/ 2.0%. The 1-year risk of major ipsilateral stroke was significantly lower in patients treated with embolus protection devices (0.0% versus 2.3%, p=0.05). CONCLUSIONS: The use of nitinol self-expanding stents in the carotid position appears to be a safe and efficacious alternative for the treatment of carotid bifurcation disease. The addition of embolus protection devices may diminish the risk of postprocedural major ipsilateral strokes. PMID- 16212453 TI - Procedure-related complications and early neurological adverse events of unprotected and protected carotid stenting: temporal trends in a consecutive patient series. AB - PURPOSE: To report procedure-related complications and neurological adverse events of unprotected over-the-wire (OTW) and protected rapid exchange (RX) carotid artery stenting (CAS) in a single-center patient series during an 8-year period. METHODS: Between 1997 and 2004, 651 consecutive patients (445 men; median age 72 years, interquartile range 64-77) were enrolled in a registry database of carotid stent procedures; from 1997 to 2002, 471 patients underwent unprotected CAS using an OTW technique, while the other 180 patients were treated with protected CAS using RX monorail systems from 2003 to 2004. Technical success and complications (neurological, hemodynamic instability, and access site) up to 30 days post intervention were analyzed. RESULTS: Technical success improved from 95% with unprotected OTW CAS to 99% with protected RX CAS (p=0.025). Procedure related complications occurred in 86 (18.3%) of 471 unprotected OTW CAS versus 18 (10.0%) of 180 protected RX CAS procedures (p=0.010). Transient ischemic attacks (3.2% versus 2.8%), minor stroke (1.7% versus 0.6%), and major stroke (2.1% versus 0.6%) showed a trend toward a difference between unprotected OTW and protected RX CAS (p=0.076); combined 30-day stroke/death rates were 3.8% for OTW versus 1.2% for RX CAS (p=0.073). During the 8-year period from 1997 to 2004, the annual rates of procedure-related complications (p=0.002), neurological events (p=0.040), and stroke and death (p=0.14) markedly decreased. CONCLUSIONS: Carotid stenting became substantially safer in the era of protected RX technology. In addition to a reduction in neurological complications, which may be due to cerebral protection devices, the improved technical success and reduced non neurological procedure-related complications are likely due to recent technical advances unrelated to cerebral protection. PMID- 16212454 TI - Endovascular treatment of ruptured abdominal aortic aneurysms: a shift of the paradigm? AB - PURPOSE: To compare endovascular aneurysm repair (EVAR) of ruptured abdominal aortic aneurysms (rAAA) to the results with open surgery. METHODS: Between May 2001 and January 2004, 50 patients were diagnosed with rAAA. Fifteen (30%) patients (14 men; median age 73 years, range 58-85) underwent EVAR, while 26 (52%) patients (23 men; median age 75 years, range 60-84) had open surgery. Nine (18%) patients (5 men; median age 86 years, range 77-91) were not operated upon. Circulatory shock was defined as systolic blood pressure<80 mmHg. Mortality was defined as death within 30 days after operation; in cases where hospital stay exceeded 30 days, in-hospital mortality was registered. Five risk factors (age>76 years, loss of consciousness, hemoglobin<90 g/L, creatinine>190 micromol/L, and electrocardiographic ischemia) were analyzed. RESULTS: In the EVAR group, 93% (14/15) of the aneurysms were excluded from the bloodstream; there were 2 (13%) conversions: 1 intraoperatively for stent-graft migration and another owing to dissection prior to hospital discharge. Mortality after open surgery was 46% (12/26) versus 13% (2/15) in the EVAR group (p>0.05). Univariate analysis without considering variables other than mortality resulted in OR 5.4 (95% CI 0.9 to 58; p=0.07). Considering risk factors and shock, multivariate analysis resulted in OR 6.5 (95% CI 0.8 to 96; p=0.08). In the EVAR group, 60% (9/15) had complications; in the group with open surgery, the complication rate was 85% (22/26; p=0.13). CONCLUSIONS: It is possible to treat rAAA with EVAR. Hypotensive patients can, at least initially, be operated under local anesthesia to stabilize blood pressure utilizing a percutaneously inserted occlusion balloon. There was a trend in our study for reduced mortality and morbidity with EVAR, but further studies are required to conclude whether EVAR significantly increases survival and reduces complications. PMID- 16212455 TI - Balloon occlusion of the aorta during endovascular repair of ruptured abdominal aortic aneurysm. AB - PURPOSE: To describe a technique of aortic clamping during endovascular aneurysm repair (EVAR) in patients with ruptured abdominal aortic aneurysms (AAA) and circulatory collapse. TECHNIQUE: A balloon catheter is inserted percutaneously from the femoral artery and inflated in the suprarenal aorta. An introducer sheath must support the balloon. The stent-graft is passed from the contralateral groin and deployed beneath the balloon. The sheath makes it possible to retrieve the balloon after the endograft has been deployed. Carbon dioxide facilitates angiography while the aortic blood flow is arrested. CONCLUSIONS: The aortic stent-graft can be deployed while the aorta is continuously "clamped" from a transfemoral approach. This may allow EVAR in patients with circulatory collapse due to aneurysm rupture. PMID- 16212456 TI - Computed tomography versus color duplex ultrasound for surveillance of abdominal aortic stent-grafts. AB - PURPOSE: To compare the ability of computed tomography (CT) and color duplex ultrasound (CDUS) to detect endoleak and accurately measure aortic aneurysm diameters after endovascular repair. METHODS: Between February 2000 and October 2004, 178 consecutive patients (156 men; mean age 74 years, range 49-89) were treated with aortic stent-grafts (86 Ancure, 55 AneuRx, and 37 Excluder). The follow-up protocol included serial CT and CDUS at 1 month and every 6 months thereafter. Sensitivity, specificity, positive predictive value, negative predictive value, and Kappa statistics (kappa) were calculated using CT as the gold standard; Bland-Altman analysis was used to determine the 95% limits of agreement. Paired and unpaired t tests and correlation coefficients were used to compare the methods. RESULTS: Follow-up ranged from 1 to 53 months (mean 16), during which 367 paired CT and CDUS studies were acquired. The mean diameter of the AAA sac after repair was 5.15 cm by CT versus 4.99 cm by CDUS (p=0.07); 93% of paired studies were somewhat similar (or=70% diameter stenosis were treated with stents selected at the discretion of the operator. Eighteen patients (32 lesions) treated with 9 different types of stents were excluded from the analysis, leaving 125 patients (69 men; mean age 67 years, range 42 to 90) with 147 lesions who received either a Radix carbofilm coated stent in 78 (53%) lesions (68 [54%] patients) or a Palmaz Genesis bare stainless steel stent in 69 (47%) lesions (57 [46%] patients). The target vessel diameter ranged from 5 to 7 mm. RESULTS: Baseline characteristics were similar in both groups except the mean stent diameter, which was larger in the bare stent cohort (6.4+/-0.7 versus 5.9+/-0.5 mm, p<0.001). Primary success was 100% in both groups; the initial mean diameter stenosis was reduced from 79%+/-14% and 80%+/ 14% in the coated versus bare stent groups to 3+/-5% and 2+/-6%, respectively. After a mean follow-up of 22+/-5 months, the restenosis rate was 6.4% for the coated stent and 5.8% for the bare stent (p=0.87). For the entire cohort, restenosis rates varied significantly (p<0.05) according to stent diameter: 19% (5/26) for 5 mm, 4% (3/81) for 6 mm, and 2.5% (1/40) for 7 mm. In a binary logistic regression analysis including carbofilm coating, stent diameter, gender, diabetes, smoking status, and body mass index, stent diameter was the only independent predictor of restenosis (odds ratio 0.12, 95% CI 0.03 to 0.48 [p=0.003] for a 1-mm increase in vessel diameter). CONCLUSIONS: Using modern low profile stent devices, carbofilm coating does not significantly reduce the restenosis rate compared to a bare metal stent. With contemporary stent devices, the restenosis rate has been decreasing compared to earlier reports in the literature. PMID- 16212463 TI - Stent fracture after endoluminal repair of a carotid artery pseudoaneurysm. AB - PURPOSE: To present a case in which a covered stent fractured 7 months after implantation in the internal carotid artery (ICA). CASE REPORT: A 59-year-old man presented with a large traumatic pseudoaneurysm of the left ICA. Seven months after successful treatment with a covered Symbiot stent, clinical complaints recurred because the pseudoaneurysm recanalized due to fracture of the stent at its midsection. Surgical stent explantation was needed, with polytetrafluoroethylene graft reconstruction of the ICA. No complications occurred in the periprocedural period. A control duplex scan 12 months later showed exclusion of the pseudoaneurysm and no graft-related complications. CONCLUSIONS: This case demonstrates an as yet unreported complication of stent grafting in a carotid artery. PMID- 16212464 TI - Endovascular treatment of an internal iliac artery aneurysm using a nitinol vascular occlusion plug. AB - PURPOSE: To report endovascular occlusion of an internal iliac artery (IIA) aneurysm with an Amplatz nitinol vascular occlusion plug. CASE REPORT: A 71-year old asymptomatic man who had previously undergone open aortic aneurysm repair presented for annual follow-up. A bifurcated Dacron graft had been inserted 12 years ago from the infrarenal aorta to the left common femoral artery and the right common iliac artery. The left common iliac artery was ligated proximally, and the left external iliac artery (EIA) provided retrograde flow into the IIA. Magnetic resonance imaging (MRI) revealed a 7.4-cm aneurysm of the left IIA. After transfemoral calibrated catheter angiography was performed, the proximal EIA was occluded with an Amplatz nitinol vascular occlusion plug. In addition, microcoils were placed distal to the vascular plug to achieve complete thrombosis of the vessel. One day after treatment, the patient was discharged free of symptoms after MRI had shown complete obliteration of the IIA aneurysm. At 6 months, the patient was free from symptoms, and angiography confirmed exclusion of the IIA aneurysm. CONCLUSIONS: This case illustrates the technical feasibility and successful short-term follow-up of a novel embolization approach to IIA aneurysms in patients with an aortofemoral graft. PMID- 16212465 TI - Re: "extension of dissection in stent-graft treatment of type B aortic dissection: lessons learned from endovascular experience". PMID- 16212466 TI - Role of multidetector computed tomography in stent-graft repair of type B aortic dissection in a patient with an aberrant right subclavian artery. PMID- 16212467 TI - Dynamic anatomy of the popliteal artery: might culture affect the outcome of endovascular therapy? PMID- 16212468 TI - "Rapid-impact interventions": how a policy of integrated control for Africa's neglected tropical diseases could benefit the poor. PMID- 16212469 TI - Diabetic retinopathy screening model for rural population: awareness and screening methodology. AB - CONTEXT: Worldwide, the prevalence of diabetic retinopathy is increasing at an alarming rate. WHO has predicted that in India the number of adults with diabetes will be the highest in the world: from 19 million in 1995 to 80 million in 2030. Although originally thought to be a disease of an urban population, the prevalence of diabetes mellitus is increasing in rural areas as well. The socioeconomic burden resulting from visual impairment or blindness caused by diabetic retinopathy, particularly in the working age group, is a serious concern. ISSUE: In order to combat diabetic retinopathy related blindness, Sankara Nethralaya, the premier eye institute of India, in collaboration with the Lions Clubs International Foundation (LCIF) and the RD Tata Trust, Mumbai, India launched a major diabetic retinopathy screening program in the rural community of South India. The objectives were to create awareness among the rural population of diabetic retinopathy with emphasis on early detection, to conduct diabetes and diabetic retinopathy screening camps, and to bring to the base hospital patients who have sight-threatening diabetic retinopathy, for ancillary investigations such as fluorescein angiography, ultrasound and to perform laser photocoagulation or vitreous surgery, or both. Other objectives included training general ophthalmologists and general physicians in order to develop an integrated diabetic retinopathy model. To address the question as to why certain individuals run the risk of developing sight threatening diabetic retinopathy, biochemical and genetic factors were also studied. The program was launched in June 2003 and 3 rural districts have been screened. To the time of writing, 128 screening camps had been organized, 103 awareness meetings conducted, 23 ophthalmologists trained and 43 general physicians attended the continuing medical education program on diabetic retinopathy. LESSONS: The key elements in the successful implementation of this program have been a team approach, involvement of community leaders and voluntary organizations, and support of the district and state administrators. PMID- 16212486 TI - Mechanisms of apoptosis through structural biology. AB - Apoptosis plays a central role in the development and homeostasis of metazoans. Research in the past two decades has led to the identification of hundreds of genes that govern the initiation, execution, and regulation of apoptosis. An earlier focus on the genetic and cell biological characterization has now been complemented by systematic biochemical and structural investigation, giving rise to an unprecedented level of clarity in many aspects of apoptosis. In this review, we focus on the molecular mechanisms of apoptosis by synthesizing available biochemical and structural information. We discuss the mechanisms of ligand binding to death receptors, actions of the Bcl-2 family of proteins, and caspase activation, inhibition, and removal of inhibition. Although an emphasis is given to the mammalian pathways, a comparative analysis is applied to related mechanistic information in Drosophila and Caenorhabditis elegans. PMID- 16212485 TI - In awe of subcellular complexity: 50 years of trespassing boundaries within the cell. AB - In this review I describe the several stages of my research career, all of which were driven by a desire to understand the basic mechanisms responsible for the complex and beautiful organization of the eukaryotic cell. I was originally trained as an electron microscopist in Argentina, and my first major contribution was the introduction of glutaraldehyde as a fixative that preserved the fine structure of cells, which opened the way for cytochemical studies at the EM level. My subsequent work on membrane-bound ribosomes illuminated the process of cotranslational translocation of polypeptides across the ER membrane and led to the formulation, with Gunter Blobel, of the signal hypothesis. My later studies with many talented colleagues contributed to an understanding of ER structure and function and aspects of the mechanisms that generate and maintain the polarity of epithelial cells. For this work my laboratory introduced the now widely adopted Madin-Darby canine kidney (MDCK) cell line, and demonstrated the polarized budding of envelope viruses from those cells, providing a powerful new system that further advanced the field of protein traffic. PMID- 16212487 TI - Regulation of protein activities by phosphoinositide phosphates. AB - Phosphoinositide phosphates (PIPs) correspond to phosphorylated derivatives of phosphatidylinositol (PI). Despite their relatively low abundance in the plasma membrane, PIPs play a crucial role as precursors of second messengers and are themselves important signaling and targeting molecules. Indeed, modulation of levels of PIPs affects, for example, cortical actin organization, membrane dynamics, and cell migration. The focus of this review is on selected interesting targets of PIPs. Those proteins that bind PIPs and are involved in regulation of actin assembly, actin membrane linkage, and actin contractility are discussed, as well as those that are involved in signaling, such as small GTPases, protein kinases, and phosphatases, or in regulation of membrane dynamics. PMID- 16212488 TI - Principles of lysosomal membrane digestion: stimulation of sphingolipid degradation by sphingolipid activator proteins and anionic lysosomal lipids. AB - Sphingolipids and glycosphingolipids are membrane components of eukaryotic cell surfaces. Their constitutive degradation takes place on the surface of intra endosomal and intra-lysosomal membrane structures. During endocytosis, these intra-lysosomal membranes are formed and prepared for digestion by a lipid sorting process during which their cholesterol content decreases and the concentration of the negatively charged bis(monoacylglycero)phosphate (BMP)- erroneously also called lysobisphosphatidic acid (LBPA)--increases. Glycosphingolipid degradation requires the presence of water-soluble acid exohydrolases, sphingolipid activator proteins, and anionic phospholipids like BMP. The lysosomal degradation of sphingolipids with short hydrophilic head groups requires the presence of sphingolipid activator proteins (SAPs). These are the saposins (Saps) and the GM2 activator protein. Sphingolipid activator proteins are membrane-perturbing and lipid-binding proteins with different specificities for the bound lipid and the activated enzyme-catalyzed reaction. Their inherited deficiency leads to sphingolipid- and membrane-storage diseases. Sphingolipid activator proteins not only facilitate glycolipid digestion but also act as glycolipid transfer proteins facilitating the association of lipid antigens with immunoreceptors of the CD1 family. PMID- 16212489 TI - Cajal bodies: a long history of discovery. AB - This review surveys what is known about the structure and function of the subnuclear domains called Cajal bodies (CBs). The major focus is on CBs in mammalian cells but we provide an overview of homologous CB structures in other organisms. We discuss the protein and RNA components of CBs, including factors recently found to associate in a cell cycle-dependent fashion or under specific metabolic or stress conditions. We also consider the dynamic properties of both CBs and their molecular components, based largely on recent data obtained thanks to the advent of improved in vivo detection and imaging methods. We discuss how these data contribute to an understanding of CB functions and highlight major questions that remain to be answered. Finally, we consider the interesting links that have emerged between CBs and alterations in nuclear structure apparent in a range of human pathologies, including cancer and inherited neurodegenerative diseases. We speculate on the relationship between CB function and molecular disease. PMID- 16212490 TI - Assembly of variant histones into chromatin. AB - Chromatin can be differentiated by the deposition of variant histones at centromeres, active genes, and silent loci. Variant histones are assembled into nucleosomes in a replication-independent manner, in contrast to assembly of bulk chromatin that is coupled to replication. Recent in vitro studies have provided the first glimpses of protein machines dedicated to building and replacing alternative nucleosomes. They deposit variant H2A and H3 histones and are targeted to particular functional sites in the genome. Differences between variant and canonical histones can have profound consequences, either for delivery of the histones to sites of assembly or for their function after incorporation into chromatin. Recent studies have also revealed connections between assembly of variant nucleosomes, chromatin remodeling, and histone post translational modification. Taken together, these findings indicate that chromosome architecture can be highly dynamic at the most fundamental level, with epigenetic consequences. PMID- 16212491 TI - Planar cell polarization: an emerging model points in the right direction. AB - Polarization is a feature common to many cell types. Epithelial cells, for example, exhibit a characteristic apical-basolateral polarity that is critical for their function. In addition to this ubiquitous form of polarity, whole fields of cells are often polarized in a plane perpendicular to the apical-basal axis. This form of polarity, referred to as planar cell polarity (PCP), exists in all adult Drosophila cuticular tissues, as well as in numerous vertebrate tissues, including the mammalian skin and inner ear epithelia. Recent advances in the study of PCP establishment are beginning to unravel the molecular mechanisms underlying this cellular process. This review discusses new developments in the molecular understanding of PCP in Drosophila and vertebrates and integrates the current data in a model to illustrate how interactions between PCP factors might function to generate planar polarity. PMID- 16212492 TI - Molecular mechanisms of steroid hormone signaling in plants. AB - Brassinosteroids (BRs), the polyhydroxylated steroid hormones of plants, regulate the growth and differentiation of plants throughout their life cycle. Over the past several years, genetic and biochemical approaches have yielded great progress in understanding BR signaling. Unlike their animal counterparts, BRs are perceived at the plasma membrane by direct binding to the extracellular domain of the BRI1 receptor S/T kinase. BR perception initiates a signaling cascade, acting through a GSK3 kinase, BIN2, and the BSU1 phosphatase, which in turn modulates the phosphorylation state and stability of the nuclear transcription factors BES1 and BZR1. Microarray technology has been used extensively to provide a global view of BR genomic effects, as well as a specific set of target genes for BES1 and BZR1. These gene products thus provide a framework for how BRs regulate the growth of plants. PMID- 16212493 TI - Anisotropic expansion of the plant cell wall. AB - Plants shape their organs with a precision demanded by optimal function; organ shaping requires control over cell wall expansion anisotropy. Focusing on multicellular organs, I survey the occurrence of expansion anisotropy and discuss its causes and proposed controls. Expansion anisotropy of a unit area of cell wall is characterized by the direction and degree of anisotropy. The direction of maximal expansion rate is usually regulated by the direction of net alignment among cellulose microfibrils, which overcomes the prevailing stress anisotropy. In some stems, the directionality of expansion of epidermal cells is controlled by that of the inner tissue. The degree of anisotropy can vary widely as a function of position and of treatment. The degree of anisotropy is probably controlled by factors in addition to the direction of microfibril alignment. I hypothesize that rates of expansion in maximal and minimal directions are regulated by distinct molecular mechanisms that regulate interactions between matrix and microfibrils. PMID- 16212495 TI - Rho GTPases: biochemistry and biology. AB - Approximately one percent of the human genome encodes proteins that either regulate or are regulated by direct interaction with members of the Rho family of small GTPases. Through a series of complex biochemical networks, these highly conserved molecular switches control some of the most fundamental processes of cell biology common to all eukaryotes, including morphogenesis, polarity, movement, and cell division. In the first part of this review, we present the best characterized of these biochemical pathways; in the second part, we attempt to integrate these molecular details into a biological context. PMID- 16212496 TI - Spatial control of cell expansion by the plant cytoskeleton. AB - The cytoskeleton plays important roles in plant cell shape determination by influencing the patterns in which cell wall materials are deposited. Cortical microtubules are thought to orient the direction of cell expansion primarily via their influence on the deposition of cellulose into the wall, although the precise nature of the microtubule-cellulose relationship remains unclear. In both tip-growing and diffusely growing cell types, F-actin promotes growth and also contributes to the spatial regulation of growth. F-actin has been proposed to play a variety of roles in the regulation of secretion in expanding cells, but its functions in cell growth control are not well understood. Recent work highlighted in this review on the morphogenesis of selected cell types has yielded substantial new insights into mechanisms governing the dynamics and organization of cytoskeletal filaments in expanding plant cells and how microtubules and F-actin interact to direct patterns of cell growth. Nevertheless, many important questions remain to be answered. PMID- 16212494 TI - RNA transport and local control of translation. AB - In eukaryotes, the entwined pathways of RNA transport and local translational regulation are key determinants in the spatio-temporal articulation of gene expression. One of the main advantages of this mechanism over transcriptional control in the nucleus lies in the fact that it endows local sites with independent decision-making authority, a consideration that is of particular relevance in cells with complex cellular architecture such as neurons. Localized RNAs typically contain codes, expressed within cis-acting elements, that specify subcellular targeting. Such codes are recognized by trans-acting factors, adaptors that mediate translocation along cytoskeletal elements by molecular motors. Most transported mRNAs are assumed translationally dormant while en route. In some cell types, especially in neurons, it is considered crucial that translation remains repressed after arrival at the destination site (e.g., a postsynaptic microdomain) until an appropriate activation signal is received. Several candidate mechanisms have been suggested to participate in the local implementation of translational repression and activation, and such mechanisms may target translation at the level of initiation and/or elongation. Recent data indicate that untranslated RNAs may play important roles in the local control of translation. PMID- 16212497 TI - RNA silencing systems and their relevance to plant development. AB - RNA silencing refers to a broad range of phenomena sharing the common feature that large, double-stranded RNAs or stem-loop precursors are processed to ca. 21 26 nucleotide small RNAs, which then guide the cleavage of cognate RNAs, block productive translation of these RNAs, or induce methylation of specific target DNAs. Although the core mechanisms are evolutionarily conserved, epigenetic maintenance of silencing by amplification of small RNAs and the elaboration of mobile, RNA-based silencing signals occur predominantly in plants. Plant RNA silencing systems are organized into a network with shared components and overlapping functions. MicroRNAs, and probably trans-acting small RNAs, help regulate development at the posttranscriptional level. Small interfering RNAs associated with transgene- and virus-induced silencing function primarily in defending against foreign nucleic acids. Another system, which is concerned with RNA-directed methylation of DNA repeats, seems to have roles in epigenetic silencing of certain transposable elements and genes under their control. PMID- 16212498 TI - Quorum sensing: cell-to-cell communication in bacteria. AB - Bacteria communicate with one another using chemical signal molecules. As in higher organisms, the information supplied by these molecules is critical for synchronizing the activities of large groups of cells. In bacteria, chemical communication involves producing, releasing, detecting, and responding to small hormone-like molecules termed autoinducers . This process, termed quorum sensing, allows bacteria to monitor the environment for other bacteria and to alter behavior on a population-wide scale in response to changes in the number and/or species present in a community. Most quorum-sensing-controlled processes are unproductive when undertaken by an individual bacterium acting alone but become beneficial when carried out simultaneously by a large number of cells. Thus, quorum sensing confuses the distinction between prokaryotes and eukaryotes because it enables bacteria to act as multicellular organisms. This review focuses on the architectures of bacterial chemical communication networks; how chemical information is integrated, processed, and transduced to control gene expression; how intra- and interspecies cell-cell communication is accomplished; and the intriguing possibility of prokaryote-eukaryote cross-communication. PMID- 16212499 TI - Pushing the envelope: structure, function, and dynamics of the nuclear periphery. AB - The nuclear envelope (NE) is a highly specialized membrane that delineates the eukaryotic cell nucleus. It is composed of the inner and outer nuclear membranes, nuclear pore complexes (NPCs) and, in metazoa, the lamina. The NE not only regulates the trafficking of macromolecules between nucleoplasm and cytosol but also provides anchoring sites for chromatin and the cytoskeleton. Through these interactions, the NE helps position the nucleus within the cell and chromosomes within the nucleus, thereby regulating the expression of certain genes. The NE is not static, rather it is continuously remodeled during cell division. The most dramatic example of NE reorganization occurs during mitosis in metazoa when the NE undergoes a complete cycle of disassembly and reformation. Despite the importance of the NE for eukaryotic cell life, relatively little is known about its biogenesis or many of its functions. We thus are far from understanding the molecular etiology of a diverse group of NE-associated diseases. PMID- 16212500 TI - Integrin structure, allostery, and bidirectional signaling. AB - Alphabeta heterodimeric integrins mediate dynamic adhesive cell-cell and cell extracellular matrix (ECM) interactions in metazoa that are critical in growth and development, hemostasis, and host defense. A central feature of these receptors is their capacity to change rapidly and reversibly their adhesive functions by modulating their ligand-binding affinity. This is normally achieved through interactions of the short cytoplasmic integrin tails with intracellular proteins, which trigger restructuring of the ligand-binding site through long range conformational changes in the ectodomain. Ligand binding in turn elicits conformational changes that are transmitted back to the cell to regulate diverse responses. The publication of the integrin alphaVbeta3 crystal structure has provided the context for interpreting decades-old biochemical studies. Newer NMR, crystallographic, and EM data, reviewed here, are providing a better picture of the dynamic integrin structure and the allosteric changes that guide its diverse functions. PMID- 16212501 TI - Centrosomes in cellular regulation. AB - Centrosomes, spindle pole bodies, and related structures in other organisms are a morphologically diverse group of organelles that share a common ability to nucleate and organize microtubules and are thus referred to as microtubule organizing centers or MTOCs. Features associated with MTOCs include organization of mitotic spindles, formation of primary cilia, progression through cytokinesis, and self-duplication once per cell cycle. Centrosomes bind more than 100 regulatory proteins, whose identities suggest roles in a multitude of cellular functions. In fact, recent work has shown that MTOCs are required for several regulatory functions including cell cycle transitions, cellular responses to stress, and organization of signal transduction pathways. These new liaisons between MTOCs and cellular regulation are the focus of this review. Elucidation of these and other previously unappreciated centrosome functions promises to yield exciting scientific discovery for some time to come. PMID- 16212502 TI - Endoplasmic reticulum-associated degradation. AB - Secretory and transmembrane proteins enter the secretory pathway through the protein-conducting Sec61 channel in the membrane of the endoplasmic reticulum. In the endoplasmic reticulum, proteins fold, are frequently covalently modified, and oligomerize before they are packaged into transport vesicles that shuttle them to the Golgi complex. Proteins that misfold in the endoplasmic reticulum are selectively transported back across the endoplasmic reticulum membrane to the cytosol for degradation by proteasomes. Depending on the topology of the defect in the protein, cytosolic or lumenal chaperones are involved in its targeting to degradation. The export channel for misfolded proteins is likely also formed by Sec61p. Export may be powered by AAA-ATPases of the proteasome 19S regulatory particle or Cdc48p/p97. Exported proteins are frequently ubiquitylated prior to degradation and are escorted to the proteasome by polyubiquitin-binding proteins. PMID- 16212503 TI - The lymphatic vasculature: recent progress and paradigms. AB - The field of lymphatic research has been recently invigorated by the identification of genes and mechanisms that control various aspects of lymphatic development. We are beginning to understand how, starting from a subgroup of embryonic venous endothelial cells, the whole lymphatic system forms in a stepwise manner. The generation of genetically engineered mice with defects in different steps of the lymphangiogenic program has provided models that are increasing our understanding of the lymphatic system in health and disease. This knowledge, in turn, should lead to the development of better diagnostic methods and treatments of lymphatic disorders and tumor metastasis. PMID- 16212504 TI - Regulation of root apical meristem development. AB - The establishment of the Angiosperm root apical meristem is dependent on the specification of a stem cell niche and the subsequent development of the quiescent center at the presumptive root pole. Distribution of auxin and the establishment of auxin maxima are early formative steps in niche specification that depend on the expression and distribution of auxin carriers. Auxin specifies stem cell niche formation by directly and indirectly affecting gene activities. Part of the indirect regulation by auxin may involve changes in redox, favoring local, oxidized microenvironments. Formation of a QC is required for root meristem development and elaboration. Many signals likely pass between the QC and the adjacent root meristem tissues. Disappearance of the QC is associated with roots becoming determinate. Given the many auxin feedback loops, we hypothesize that roots evolved as part of an auxin homeostasis mechanism. PMID- 16212505 TI - Phagocytosis: at the crossroads of innate and adaptive immunity. AB - Phagocytosis, the process by which cells engulf large particles, requires a substantial contribution of membranes. Recent studies have revealed that intracellular compartments, including endocytic organelles and the endoplasmic reticulum (ER), can engage in fusion events with the plasma membrane at the sites of nascent phagosomes. The finding that ER proteins are delivered to phagosomes, where degraded peptides are loaded onto major histocompatibility complex (MHC) class II molecules, has significantly enhanced our understanding of the immune functions associated with these organelles. Although it is well known that pathogens are killed in phagosomes, the contribution of ER proteins to phagosomes has provided a novel pathway for the loading of exogenous peptides onto MHC class I molecules, a process known as cross-presentation. Thus, phagocytosis has evolved from a nutritional function in unicellular organisms to play key roles in both innate and adaptive immunity in vertebrates. PMID- 16212506 TI - Protein translocation by the Sec61/SecY channel. AB - The conserved protein-conducting channel, referred to as the Sec61 channel in eukaryotes or the SecY channel in eubacteria and archaea, translocates proteins across cellular membranes and integrates proteins containing hydrophobic transmembrane segments into lipid bilayers. Structural studies illustrate how the protein-conducting channel accomplishes these tasks. Three different mechanisms, each requiring a different set of channel binding partners, are employed to move polypeptide substrates: The ribosome feeds the polypeptide chain directly into the channel, a ratcheting mechanism is used by the eukaryotic endoplasmic reticulum chaperone BiP, and a pushing mechanism is utilized by the bacterial ATPase SecA. We review these translocation mechanisms, relating biochemical and genetic observations to the structures of the protein-conducting channel and its binding partners. PMID- 16212507 TI - Retinotectal mapping: new insights from molecular genetics. AB - The sensory and motor components of nervous systems are connected topographically and contain neural maps of the external world. The paradigm for such maps is the precisely ordered wiring of the output cells of the eye to their synaptic targets in the tectum of the midbrain. The retinotectal map is organized in development through the graded activity of Eph receptor tyrosine kinases and their ephrin ligands. These signaling proteins are arrayed in complementary expression gradients along the orthogonal axes of the retina and tectum, and provide both input and recipient cells with Cartesian coordinates that specify their location. Molecular genetic studies in the mouse indicate that these coordinates are interpreted in the context of neuronal competition for termination sites in the tectum. They further suggest that order in the retinotectal map is determined by ratiometric rather than absolute difference comparisons in Eph signaling along the temporal-nasal and dorsal-ventral axes of the eye. PMID- 16212508 TI - In vivo imaging of lymphocyte trafficking. AB - Over the past decades, intravital microscopy (IVM), the imaging of cells in living organisms, has become a valuable tool for studying the molecular determinants of lymphocyte trafficking. Recent advances in microscopy now make it possible to image cell migration and cell-cell interactions in vivo deep within intact tissues. Here, we summarize the principal techniques that are currently used in IVM, discuss options and tools for fluorescence-based visualization of lymphocytes in microvessels and tissues, and describe IVM models used to explore lymphoid and non-lymphoid organs. The latter will be introduced according to the physiologic itinerary of developing and differentiating T and B lymphocytes as they traffic through the body, beginning with their development in bone marrow and thymus and continuing with their migration to secondary lymphoid organs and peripheral tissues. PMID- 16212509 TI - Stem cell niche: structure and function. AB - Adult tissue-specific stem cells have the capacity to self-renew and generate functional differentiated cells that replenish lost cells throughout an organism's lifetime. Studies on stem cells from diverse systems have shown that stem cell function is controlled by extracellular cues from the niche and by intrinsic genetic programs within the stem cell. Here, we review the remarkable progress recently made in research regarding the stem cell niche. We compare the differences and commonalities of different stem cell niches in Drosophila ovary/testis and Caenorhabditis elegans distal tip, as well as in mammalian bone marrow, skin/hair follicle, intestine, brain, and testis. On the basis of this comparison, we summarize the common features, structure, and functions of the stem cell niche and highlight important niche signals that are conserved from Drosophila to mammals. We hope this comparative summary defines the basic elements of the stem cell niche, providing guiding principles for identification of the niche in other systems and pointing to areas for future studies. PMID- 16212510 TI - Docosahexaenoic acid, fatty acid-interacting proteins, and neuronal function: breastmilk and fish are good for you. AB - In contrast to other tissues, the nervous system is enriched in the polyunsaturated fatty acids (PUFAs): arachidonic acid (AA, 20:4 n-6) and docosahexaenoic acid (DHA, 22:6 n-3). Despite their abundance in the nervous system, AA and DHA cannot be synthesized de novo by mammals; they, or their precursors, must be ingested from dietary sources and transported to the brain. During late gestation and the early postnatal period, neurodevelopment is exceptionally rapid, and substantial amounts of PUFAs, especially DHA, are critical to ensure neurite outgrowth as well as proper brain and retina development. Here, we review the various functions of DHA in the nervous system, the proteins involved in its internalization and metabolism into phospholipids, and its relationship to several neurological disorders, including Alzheimer's disease and depression. PMID- 16212511 TI - Specificity and versatility in tgf-beta signaling through Smads. AB - The TGF-beta family comprises many structurally related differentiation factors that act through a heteromeric receptor complex at the cell surface and an intracellular signal transducing Smad complex. The receptor complex consists of two type II and two type I transmembrane serine/threonine kinases. Upon phosphorylation by the receptors, Smad complexes translocate into the nucleus, where they cooperate with sequence-specific transcription factors to regulate gene expression. The vertebrate genome encodes many ligands, fewer type II and type I receptors, and only a few Smads. In contrast to the perceived simplicity of the signal transduction mechanism with few Smads, the cellular responses to TGF-beta ligands are complex and context dependent. This raises the question of how the specificity of the ligand-induced signaling is achieved. We review the molecular basis for the specificity and versatility of signaling by the many ligands through this conceptually simple signal transduction mechanism. PMID- 16212512 TI - The great escape: when cancer cells hijack the genes for chemotaxis and motility. AB - The combined use of the new technologies of multiphoton-based intravital imaging, the chemotaxis-mediated collection of invasive cells, and high sensitivity expression profiling has allowed the correlation of the behavior of invasive tumor cells in vivo with their gene expression patterns. New insights have resulted including a gene expression signature for invasive cells and the tumor microenvironment invasion model. This model proposes that tumor invasion and metastasis can be studied as a problem resembling normal morphogenesis. We discuss how these new insights may lead to a better understanding of the molecular basis of the invasive behavior of tumor cells in vivo, which may result in new strategies for the diagnosis and treatment of metastasis. PMID- 16212513 TI - Comparative effectiveness of total population versus disease-specific neural network models in predicting medical costs. AB - The objective of this research was to compare the accuracy of two types of neural networks in identifying individuals at risk for high medical costs for three chronic conditions. Two neural network models-a population model and three disease-specific models-were compared regarding effectiveness predicting high costs. Subjects included 33,908 health plan members with diabetes, 19,264 with asthma, and 2,605 with cardiac conditions. For model development/ testing, only members with 24 months of continuous enrollment were included. Models were developed to predict probability of high costs in 2000 (top 15% of distribution) based on 1999 claims factors. After validation, models were applied to 2000 claims factors to predict probability of high 2001 costs. Each member received two scores-population model score applied to cohort and disease model score. Receiver Operating Characteristic (ROC) curves compared sensitivity, specificity, and total performance of population model and three disease models. Diabetes specific model accuracy, C = 0.786 (95%CI = 0.779-0.794), was greater than that of population model applied to diabetic cohort, C = 0.767 (0.759-0.775). Asthma specific model accuracy, C = 0.835 (0.825-0.844), was no different from that of population model applied to asthma cohort, C = 0.844 (0.835-0.853). Cardiac specific model accuracy, C = 0.651 (0.620-0.683), was lower than that of population model applied to cardiac cohort, C = 0.726 (0.697-0.756). The population model predictive power, compared to the disease model predictive power, varied by disease; in general, the larger the cohort, the greater the advantage in predictive power of the disease model compared to the population model. Given these findings, disease management program staff should test multiple approaches before implementing predictive models. PMID- 16212515 TI - Using visual displays as a tool to demonstrate disease management program effectiveness. AB - With the rapid introduction of new medical and information technologies, there are much more data available today than ever before. Translating data into useful information for a wide variety of audiences is a challenge for health care in general and disease management programs in particular. This paper addresses these issues by introducing several visual displays that illustrate important data elements in an unencumbered fashion. Examples are provided using the various stages of a hypothetical congestive heart failure (CHF) disease management program (ie, patient identification, program enrollment, intervention process, and outcomes evaluation). PMID- 16212514 TI - Patient satisfaction measurement in the disease management industry. AB - In mid-2004, the Disease Management Association of America (DMAA) Patient Satisfaction Workgroup in association with J.D. Power and Associates (JDPA) conducted a literature review and a member survey to gain an understanding of the nature of patient satisfaction measurement as it pertains to disease management (DM) programs within the DM industry. A review of the relevant literature indicates that perhaps, with the exception of diabetes disease management, there are no prevalent, systematic, or statistically validated approaches for measuring patient satisfaction within the disease management industry. Most existing studies tend to focus on the effectiveness of a disease management program on clinical outcomes, with patient satisfaction measured only as a part of a battery of "outcome" measures. However, many of these studies do find positive associations between patient satisfaction and clinical outcomes. A majority of the 49 respondents who completed the member feedback survey hold relatively high positions in their organizations. The vast majority of respondents indicate their organizations conduct patient satisfaction surveys that assess overall satisfaction, satisfaction with materials and information provided, and with staff members. Patient satisfaction surveys are most common among the five common chronic diseases: diabetes, asthma, congestive heart failure (CHF), coronary artery disease (CAD), and chronic obstructive pulmonary disease (COPD). More than three in four respondents agree that patient satisfaction measurement is important to the long-term success of their programs. Respondents also indicate that along with intelligence on patients' overall satisfaction with the program, they would also like to gain an understanding of whether or not their programs actually help manage the patient's medical condition. Eight survey instruments currently in use and submitted by study participants were also reviewed. Most of these instruments are relatively short and basic, typically administered by mail, and vary in the types of questions and response categories presented to respondents. This research concludes that there exists an implied need for patient satisfaction measurement in the DM industry and an opportunity to develop and leverage a standardized measurement approach assessing patient satisfaction. Additionally, the authors suggest that there may be value to conceptualizing "patient satisfaction" not as an outcome in itself, but as a means to increase compliance, which, in turn, can improve medical outcomes. PMID- 16212516 TI - Outcomes of American Lung Association-Indiana Lung Centers asthma program. AB - The American Lung Association of Indiana (ALA-I), in conjunction with participating Indiana hospitals, developed the Lung Center concept as a mechanism to provide standardized delivery of lung health education. The goal of this pilot study was to evaluate initial experience with the Lung Center program "Overcoming Your Asthma," a two-session asthma education program, and identify areas needing improvement. A total of 305 participants responded to a 31-item questionnaire at baseline (immediately prior to program exposure) and again at 1 month (n = 75) and 6 months (n = 30) after participation. Overall, delivery of the ALA-I Lung Center asthma education program improved respondents' experience with asthma. At one month after the educational session, the program improved participant knowledge about asthma. This was associated with modest improvements in treatment behaviors, economic outcomes and asthma symptoms such as reduced breathing difficulties, wheezing and asthma exacerbations, and improvement in sleep. Improvements were not uniformly sustained at 6 months. In summary, the Lung Center asthma education program appears to benefit patients with asthma. The results provide preliminary evidence to support continued delivery of asthma education in Lung Centers. Future efforts should emphasize education to improve treatment attitudes and behaviors. PMID- 16212517 TI - Helping patients improve their health-related behaviors: what system changes do we need? AB - A large gap separates actual clinical outcomes of patients with chronic disease from optimal outcomes. This gap may be attributable to physician problems (physicians unaware of practice guidelines), patient problems (patients who choose not to follow medical advice) or system problems (physicians lack time to assist patients in managing their chronic condition). A goal for chronic illness care is to assist people with chronic conditions to become informed, activated patients. Patients who are informed and activated (activated = participating in health-related decisions) have better health-related behaviors and clinical outcomes. Primary care is often conducted within a 15-minute, multi-agenda visit between physician and patient. Such a structure has led to (1) patients being inadequately informed about their chronic conditions and (2) patients being passive recipients of medical advice rather than active participants in medical decisions. The result has been poorly informed, passive patients. This situation constitutes a system problem. Three redesign elements have the potential to address this system problem: (1) Pre-activating patients prior to the clinical visit appears to encourage more active patients and--in one study--improved clinical outcomes. (2) Planned visits, with a care manager spending time with patients, individually or in groups, providing education and medical management, have been shown to improve clinical outcomes. (3) Regular sustained follow-up, by face-to-face visits, telephone, or electronic means, is associated with healthier behaviors. Not all patients receiving disease management through redesigned primary care will adopt healthier behaviors because many factors outside the medical care system influence personal choices. However, until the medical care system regularly offers adequate information and encourages collaborative decision-making, it is improper to place the responsibility for unhealthy behaviors onto patients. PMID- 16212520 TI - Regulation of microbial immunity: the suppressor cell renaissance. AB - Regulatory/suppressor cells have become rehabilitated and respectable. They returned as regulators of autoimmunity, but are now acknowledged to critically influence immunity to foreign antigens, such as those found on microbes. This review describes the principal types of regulatory cells that influence immunity to microbes, focusing on viruses. We discuss both the merits and downside of the Treg response during infection. The mechanisms by which Treg are induced, recognize microbes, and exert their function is also discussed. Finally, we examine approaches that might prove useful to manipulate regulatory cell response against infections. PMID- 16212521 TI - Dendritic cell biology in herpesvirus infections. PMID- 16212522 TI - Viral interference with antigen presentation to CD8+ T cells: lessons from cytomegalovirus. AB - Cytomegaloviruses (CMV), in common with other Herpesviruses, establish lifelong persistence in their hosts. These highly host-specific viruses each encode viral genes that interfere with antigen presentation to CD8+ T cells, although the molecular mechanisms by which this end is achieved differ for human and murine CMVs. In each case, there has been a presumption that these genes are necessary for virus persistence in the host. However, recent data in the murine model casts doubt on that presumption. Here, we review the molecular mechanisms of interference with the class I pathway, and the published data regarding functional significance, with a focus on the murine model. PMID- 16212523 TI - Immune mechanisms in murine gammaherpesvirus-68 infection. AB - The murine gamma-herpesvirus-68 (MHV-68) is a relative of the Kaposi's sarcoma associated herpesvirus (KSHV) and Epstein-Barr virus (EBV) that infects mice. All these gamma-herpesviruses are subject to immune control, but limit the impact of this control through immune evasion. Molecular evasion mechanisms have been described in abundance. However, we can only speculate what EBV and KSHV immune evasion contributes to the viral lifecycle. With MHV-68, we can analyze in vivo the contribution of immunological and virological gene expression to pathogenesis. While the physiology of infection seems quite well conserved between these viruses, the pathologies associated with immune suppression are obviously very different. MHV-68 is therefore more suited to uncovering the basic biology of gamma-herpesvirus infection than to testing disease interventions. Nevertheless, it may make some useful predictions about effective strategies of vaccination and infection control. This review aims to outline our current state of knowledge and to highlight some limitations of the MHV-68 model as it stands, in the hope of stimulating constructive progress. PMID- 16212524 TI - Herpes simplex virus and toll-like receptors. AB - The earliest interactions between viruses and host cells are critical to determining the outcome of infections. The interactions between a virus and its host cells that lead to the production of inflammatory cytokines are essential to the development of T cells and antibodies that provide long-term defense against the invaders. At the same time, however, these same cytokines (or sometimes other mediators or chemokines) stimulate inflammation, which may lead to localized tissue damage and/or systemic circulatory collapse and death. This review focuses on the interaction between Toll-like receptors, a critical component of the innate immune system of mammals, and their role in herpes simplex pathogenesis and immunity. PMID- 16212525 TI - Immune control of HSV-1 latency. AB - A hallmark of the herpes family of viruses is their ability to cause recurrent disease. Upon primary infection, Herpes Simplex virus (HSV) establishes a latent infection in sensory neurons that persists for the life of the individual. Reactivation of these latent viral genomes with virion formation is the source of virus for most HSV recurrent disease. This review details recent exciting findings supporting a role for the host immune system, particularly CD8+ T cells in maintaining HSV-1 in a latent state. PMID- 16212526 TI - Statins could be used to control replication of some viruses, including HIV-1. AB - Statins are mainly known for their plasma cholesterol-lowering properties and are widely used for the prevention of cardiovascular diseases. They however also exert pleiotropic effects through a variety of mechanisms, among which several immunosuppressive effects that are unrelated to their cholesterol-lowering activity. Interestingly, there has been recent evidence of antiviral effects, including preliminary studies on the efficacy of statins against HIV-1. This paper more particularly focuses on the specific inhibition of the binding of leukocyte function-associated antigen-1 (LFA-1) to intercellular adhesion molecule (ICAM-1) by statins, independently of the inhibition of HMGCoA reductase. Targeting the statin-binding site within LFA-1 or regulating LFA-1 affinity by inhibiting prenylation of the small GTPases could prove useful to treat inflammatory, autoimmune diseases and possibly viral infections, including HIV-1. PMID- 16212528 TI - HLA-B polymorphism in Japanese HIV-1-infected long-term surviving hemophiliacs. AB - Approximately 30% of patients with hemophilia in Japan were infected with human immunodeficiency virus (HIV) in early 1980s through contaminated blood products. In 1995, a cohort of HIV-infected, asymptomatic patients with hemophilia was set up for follow-up study. Although the patients met the criteria for long-term non progressor (LTNP) at the entry to the cohort, some of them later developed lymphopenia during five more years of observation. We collected blood samples from 80 long-term survivors; 42 of them did not require antiviral therapy, but the rest were under treatment. Analysis of HLA-B genotype revealed that carriers of known HIV-resistant alleles such as HLA-B*5701, B*5801, and alleles of B27 antigenic group were not increased in frequency, but that HLA-B*1507 was increased in the cohort (6.25% vs. 1.03%, OR = 6.40, p = 0.039). We also observed the decrease in carriers of HLA-B*5401 (3.75% vs. 14.95%, OR = 0.22, p = 0.016). HLAB* 5401 is a relatively common allele in East Asian populations and belongs to the same B22 antigenic group as B55 and B56 which were reported to associate with rapid progression. Our data indicated that HLA class I is one of the host factors involved in the retardation of HIV disease progression as also reported in the previous studies; however, the alleles associated with this resistance were not the same because of divergent host genetic background. PMID- 16212527 TI - Gammadelta lymphocyte response to porcine reproductive and respiratory syndrome virus. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be one of the most important diseases facing swine industry today. Following PRRSV infection pigs develop both humoral and cell-mediated responses following PRRSV exposure; however, the relative importance in protection and clearance of the virus is not yet completely understood. Swine contain a large percentage of gammadelta T-lymphocytes in peripheral circulation capable of responding to various pathogens in both an innate and specific immune response. The objectives of this study were to determine whether gammadelta lymphocytes functionally respond to PRRSV upon initial exposure and re-exposure. Four month old PRRSV free gilts were intranasally inoculated with a field isolate MN-30100 then assessed at various time points post infection. On day 120, pigs were re-exposed with MN 30100 PRRSV strain and subsequently were bled on days 0, 7, and 14 post re exposure. Lymphocyte subpopulations, antigen specific proliferation, and IFN gamma production were evaluated throughout the study. Circulating gammadelta lymphocytes in PRRSV exposed animals expanded between days 14 to 70 (d14-d70, p = 0.016); following antigen stimulation, gammadelta lymphocyte proliferated by day 14 (d0-d14, p = 0.001) continuing through day 60. gammadelta lymphocytes produced IFN-gamma by day 14 pi continuing through day 50 (d0-d50, p = 0.004). Following re-exposure both gammadelta+ and CD4+ lymphocytes increased in IFN-gamma production. These results are not fully conclusive on the role of gammadelta lymphocytes against PRRSV; the data indicate that gammadelta lymphocytes specifically respond to PRRSV. PMID- 16212529 TI - In vitro susceptibility of macrophages to porcine reproductive and respiratory syndrome virus varies between genetically diverse lines of pigs. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be responsible for financial losses in the swine industry worldwide. It remains undetermined whether genetic variability of the host in susceptibility to PRRSV exists and if this variability can be exploited to help control this important disease. The objective of this study was to determine if an in vitro flow cytometry (FACS) assay that detects the percentage of monocyte-derived macrophages (MDM) infected with PRRSV could be utilized to demonstrate genetic variability in the susceptibility between distinct lines of pigs. Over 400 growing pigs from six genetic lines maintained in a single commercial breeding herd were screened using an in vitro FACS assay. From this initial screening, two genetically diverse lines of pigs that were also divergent in their FACS results were selected for further study. An additional 264 pigs from these two lines were subsequently tested for in vitro susceptibility to PRRSV. As in the preliminary screening, the Large White line had significantly higher average percent positive MDM over the Duroc-Pietrain synthetic line. This report suggests a genetic component for susceptibility to PRRSV exists and that the in vitro assay may be useful in predicting the relative susceptibility to PRRSV in large groups of animals. PMID- 16212530 TI - Expression of leukocyte immunoglobulin-like receptors and natural killer receptors on virus-specific CD8+ T cells during the evolution of Epstein-Barr virus-specific immune responses in vivo. AB - Antigen-primed cytotoxic T lymphocytes (CTL) may express leukocyte immunoglobulin like receptors (LILRs) and natural killer receptors (NKRs). Published work suggests that expression of some of these receptors confers survival advantage, leading to the idea that cells expressing such receptors may accumulate as an antigen-specific response evolves. Here we tested this hypothesis by analyzing expression of CD85j (also known as LILRB1 or ILT2), KIRs, CD94, and CD161 by Epstein- Barr virus (EBV)-specific CTL during the primary and persistent phases of EBV infection in humans. During primary infection, few EBV-specific CTL expressed these receptors and this proportion was equally low in early persistent infection. Thus, expression of these molecules does not influence capacity to survive downregulation of the primary response. However, in donors persistently infected with EBV for many years, a significantly higher proportion of EBV specific CTL expressed CD85j and NKRs, suggesting that cells expressing these receptors can accumulate with time. Using FACS analysis, we confirmed, at a single cell level, that expression of CD85j, defined by staining with the antibody VMP55, was associated with reduced capacity of EBV-specific CD8+ T cells to respond to antigen. Thus, in the later stages of persistent infection, protective immunity to EBV may be reduced due to the preferential accumulation of hyporesponsive EBV-specific CD8+ T cells. PMID- 16212531 TI - Human parainfluenza virus 3 neuraminidase activity contributes to dendritic cell maturation. AB - Mechanisms of dendritic cells (DCs) immunomodulation by parainfluenza viruses have not been characterized. We analyzed whether the human parainfluenza 3 (HPF3) virus hemagglutinin-neuraminidase glycoprotein (HN) might influence DC maturation. HN possesses a receptor binding function and a neuraminidase or desialidating activity. To assess whether the neuraminidase activity of HN affects DC maturation, human myeloid DCs were exposed to either live or UV inactivated HPF3 viruses containing wild type or a mutated form of HN with decreased neuraminidase activity. Exposure of human DCs to either UV-inactivated or live virus induced up-regulation of CD83 and CD86 surface markers, morphological changes, and a cytokine expression pattern consistent with maturation. However, the level of maturation was found to be lower in DCs infected with the neuraminidase deficient variant as compared to the wild type. These results suggest that during the course of viral infection, HN's neuraminidase activity may play an important role contributing to maturation and activation of DCs. PMID- 16212532 TI - Visualization and microscopic quantification of HCMV-peptide-specific cytotoxic T lymphocytes using tetramer binding. AB - To monitor the frequencies of virus-specific cytotoxic T lymphocytes (CTLs), FACS analyses were performed detecting lymphocyte-specific surface molecules and tetramer binding, as marker for peptide-specificity. Aim of this investigation was to establish an alternative protocol for the quantification of virus-specific CTLs using tetramer binding and microscopic analyzing. The frequencies of HCMV pp65-peptide-specific CTLs in the blood of eight different HLA-A*0201-positive, HCMV-IgG antibody-positive donors were analyzed with both methods. Using FACS analyses, a median of 0.8% and, using the microscopic analyses, a median of 3.0% was detected in the CD3+CD8+ cells. After enrichment of HCMV-pp65-peptide specific CTLs using the interferon-gamma secretion assay followed by expansion in cell culture, a median of 90.6% using FACS analyses and a median of 87.1% using the microscopic analyses was detected. Thus, the staining protocol presented in this investigation is an alternative approach to detect and to quantify virus specific CTLs in low as well as in high frequencies. PMID- 16212533 TI - DNA prime followed by protein boost enhances neutralization and Th1 type immunity against FMDV. AB - Prime-boost strategy has been exhibited its potency to enhance immune responses, which would be important to the success to develop a vaccine against the foot-and mouth disease virus (FMDV). An eukaryotic expression construct encoding the FMDV capsid VP1 protein with a recombinant VP1 protein or a commercial FMDV vaccine were tested in the prime-boost strategy in mice and cattle trials. The levels of induced specific antibodies, T cell proliferations, and DTH activities were significantly higher in the prime-boost groups than in those vaccinated with DNA, protein or FMDV vaccine alone. More importantly, the levels of neutralizing antibodies in the former groups were significantly higher than others and could last for at least four months in cattle trials. This study suggests that the prime-boost strategy significantly improves the effective immunity and may provide a longer protection against FMDV infection. PMID- 16212534 TI - Identification of neutralizing epitopes on the VP2 protein of infectious bursal disease virus by phage-displayed heptapeptide library screening and synthetic peptide mapping. AB - Infectious bursal disease virus (IBDV) is the causative agent of infectious bursal disease, which is one of the most important and widespread infectious diseases in commercial chickens. Conformational epitopes have been reported in the highly variable region of the VP2 protein of IBDV. In the present study, a random heptapeptide library was screened by using monoclonal antibodies (mAbs), YNW17 and YNW29, directed to the VP2 of IBDV and two peptide motifs, D-X-P-R and A-R-G, were identified. The motifs are present on the N and C terminal sequences of the highly variable region of VP2. Synthetic overlapping peptides covering the motifs on VP2 were analyzed by Dot- ELISA with the mAbs and two epitopes 197CDSSDRPRVYTIT209 and 329ARGSLAVTI337 identified. The above epitopes were also recognized by chicken anti-IBDV sera and shown to inhibit the binding of their mAbs to recombinant VP2. Both mAbs and sera from mice immunized with the conjugated epitope-peptides were able to neutralize serotype I IBDV. These results indicated that the epitopes are two neutralizing linear B-cell epitopes and would be useful for the development of peptide-based IBD vaccines. PMID- 16212535 TI - Neutralizing monoclonal antibodies against Russian strain of the West Nile virus. AB - We have developed a panel of 16 hybridomas secreting neutralizing monoclonal antibodies (Nt- MAbs) to Russian isolate (LEIV-Vlg99-27889-human) of the West Nile virus (WNV). Most of the Nt-Mabs were either IgG1 or IgG3 subtypes. Nine of the 16 neutralizing MAbs detected WNV protein E in Western blot. According to their Nt-activities, Western blot results and cross-reactivity, the MAbs were divided into four groups. Monoclonal antibodies from group I were able to neutralize WNV strains Vlg99-27889, Vlg00-27924, Hp-94, A-1640, A-72, Tur-2914, and Eg101. The Nt-activity of MAbs from groups II-IV towards these WNV strains was variable. Recombinant fragments E(1-180), E(1-321), and E(260-466) of protein E were used for preliminary mapping of domains recognized by Nt-MAbs. Only five Nt-MAbs were able to react with the recombinant polypeptides. The MAbs 9E2, 7G9, 11G3, and 7E6 from group Ia recognized Nt-epitope(s) between amino acids 321 and 466 of protein E and Nt-MAb 4F11 (group III) reacted with residues 1-180. This demonstrates that two discrete regions of protein E are involved in neutralization of WNV. Our data on immunochemical, biological activities of Nt MAbs and mapping of Nt-epitopes using recombinant polypeptides suggest at least 13 different Nt-epitopes for WNV. PMID- 16212537 TI - Viral rebound syndrome in two HIV-1-positive patients after structured treatment interruption. PMID- 16212536 TI - Detection of infectious bronchitis virus and specific anti- viral antibodies using a Concanavalin A-Sandwich-ELISA. AB - Concanavalin A-Sandwich ELISA (Con A-S-ELISA) was developed for the detection of infectious bronchitis virus (IBV) or chicken specific anti-viral antibodies. The antigen detection limit for the Con A-S-ELISA was 10(5,1) EID(50)/mL. Three homologous and four heterologous IBV strains were similarly detected. This assay was highly effective in detecting the virus after infected tissue homogenates were passed once in embryonated chicken eggs, showing a good agreement with virus isolation technique. The Con A-S-ELISA was also used to measure anti-IBV chicken antibodies and showed a high coefficient of correlation (r = 0.85) and an agreement of k = 0.80 with the commercially available Indirect-ELISA. The relative sensitivity and specificity between these two tests were, respectively, 92.86% and 95.65% with an accuracy of 93.39%. Thus, the Con A-S-ELISA proved to be able to detect alternatively homologous and heterologous IBV strains or specific chicken anti- IBV antibodies, using the Con A as capture reagent of this assay. PMID- 16212538 TI - Quantifying structural and functional restraints on amino acid substitutions in evolution of proteins. AB - One of Oleg Ptitsyn's most important papers (Shakhnovich, E., Abkevich, V., and Ptitsyn, O. (1996) Nature, 379, 96-98) describes how knowledge of structure and function can be used to understand better the nature of amino acid substitutions in families and superfamilies of proteins. The selective advantages of retaining structure and function during evolution can be expressed as restraints on the amino acid substitutions that are accepted. PMID- 16212539 TI - Molecular mechanisms of regulation of functional activity of mononuclear phagocytes by leptin. AB - Leptin is a peptide hormone synthesized by adipocytes. The main function of leptin is associated with regulation of the body energetic balance and restriction of excess accumulation of fat. This review considers in detail the involvement of leptin in regulation of fundamental effector functions of mononuclear phagocytes, which express receptors for this hormone. Possible molecular mechanisms of modulation by leptin of phagocytic activity, oxygen dependent microbicidity, and nitric oxide generation by mononuclear phagocytes are analyzed, as well as the role of leptin in the formation of the produced cytokine pattern. The data presented suggest that the regulation of mononuclear phagocytes by leptin is associated with activation of the JAK/STAT signaling pathway, which leads to stimulation of phagocytosis, production of oxygen and nitrogen reactive species, and also to increase in secretion of pro-inflammatory cytokines. PMID- 16212540 TI - Effect of mutation of the conservative glycine residues Gly100 and Gly147 on stability of Escherichia coli inorganic pyrophosphatase. AB - Sequence alignment of inorganic pyrophosphatases (PPases) isolated from the different organisms shows that glycine residues Gly100 and Gly147 are conservative. These residues are located in flexible segments of a polypeptide chain that have similar structure in the different PPases. To elucidate the possible role of these segments in the functioning of PPase, the mutant variants Gly100Ala and Gly147Val in conservative loops have been obtained. In this work, the influence of these mutations on stability of PPase globular structure has been studied. Differential scanning calorimetry has been used to determine the apparent enthalpy of thermal denaturation for the native PPase and its mutant variants Gly100Ala and Gly147Val. Guanidine hydrochloride-induced chemical denaturation of PPase has also been studied. It is shown that the substitutions of Gly100 and Gly147 result in overall destabilization of the globular structure. PMID- 16212541 TI - Substitutions of glycine residues Gly100 and Gly147 in conservative loops decrease rates of conformational rearrangements of Escherichia coli inorganic pyrophosphatase. AB - Escherichia coli inorganic pyrophosphatase (PPase) is a one-domain globular enzyme characterized by its ability to easily undergo minor structure rearrangements involving flexible segments of the polypeptide chain. To elucidate a possible role of these segments in catalysis, catalytic properties of mutant variants of E. coli PPase Gly100Ala and Gly147Val with substitutions in the conservative loops II and III have been studied. The main result of the mutations was a sharp decrease in the rates of conformational changes required for binding of activating Mg2+ ions, whereas affinity of the enzyme for Mg2+ was not affected. The pH-independent parameters of MgPP(i) hydrolysis, kcat and kcat/Km, have been determined for the mutant PPases. The values of kcat for Gly100Ala and Gly147Val variants were 4 and 25%, respectively, of the value for the native enzyme. Parameter kcat/Km for both mutants was two orders of magnitude lower. Mutation Gly147Val increased pH-independent Km value about tenfold. The study of synthesis of pyrophosphate in the active sites of the mutant PPases has shown that the maximal level of synthesized pyrophosphate was in the case of Gly100Ala twofold, and in the case of Gly147Val fivefold, higher than for the native enzyme. The results reported in this paper demonstrate that the flexibility of the loops where the residues Gly100 and Gly147 are located is necessary at the stages of substrate binding and product release. In the case of Gly100Ala PPase, significant impairment of affinity of enzyme effector site for PP(i) was also found. PMID- 16212542 TI - Structural studies on pectin from marsh cinquefoil Comarum palustre L. AB - Pectin with [alpha]D(20) +192 degrees (c 0.1; water), named comaruman, was isolated from marsh cinquefoil Comarum palustre L., which is widespread in the European North. The sugar chain of comaruman contains residues of D-galacturonic acid (64%), D-galactose (13%), L-rhamnose (12%), L-arabinose (6%), and trace amounts of xylose and glucose. Partial acid hydrolysis and digestion with pectinase demonstrated that comaruman composed of the backbone comprised regions of linear alpha-1,4-D-galactopyranosyl uronan interconnected by numerous residues of alpha-1,2-L-rhamnopyranose. In addition to the backbone (core of the macromolecule), ramified regions are involved in comaruman and comprise alpha-2,4 L-rhamno-alpha-4-D-galacturonan with side chains consisting mainly of beta-1,4 linked residues of D-galactopyranose. The ramified region contains additionally residues of 5-O-substituted arabinofuranose and 3- and 6-O-substituted galactopyranose. The present 3,4- and 4,6-di-O-substituted residues of galactopyranose appear to be branching points of the side chains. Some galactopyranose residues were found to occupy the terminal positions of the side chains or appeared to be single sugar residues attached to the side chains. Methylation analysis data indicated that comaruman contains residues of terminal, 3- and 3,4-di-O-substituted galactopyranosyl uronic acid, which appeared to be constituents of the side chains, and the latter represented additionally branching points of the backbone. PMID- 16212544 TI - Selection and characterization of phage miniantibodies to actins of different origin. AB - Single-chain miniantibodies (scFv's) to actin were obtained by the phage display method. A naive combinatorial phage display library of murine scFv's (containing 2x10(8) independent recombinant clones) was used to select miniantibodies. After three rounds of selection two clones producing miniantibodies to chicken smooth muscle actin with affinity constants of 1.4x10(7) and 1.2x10(6) M(-1) were chosen. The isolated miniantibodies could specifically detect various plant and animal actins. PMID- 16212543 TI - Cloning, purification, and characterization of diaminobutyrate acetyltransferase from the halotolerant methanotroph Methylomicrobium alcaliphilum 20Z. AB - L-2,4-Diaminobutyrate (DAB) acetyltransferase (DABAcT) catalyzes one of the key reactions of biosynthesis of the bacterial osmoprotectant ectoine--acetylation of L-2,4-DAB yielding Ngamma-acetyl-2,4-DAB. Gene ectA encoding DABAcT was cloned from DNA of the halotolerant methanotroph Methylomicrobium alcaliphilum 20Z and expressed in Escherichia coli with an additional six His residues at the C terminus. Homogeneous enzyme preparation with specific activity 200 U/mg was obtained by affinity metal-chelating chromatography. DABAcT was found to be a homodimer with molecular mass 40 kD. The enzyme is most active at pH 9.5 and 20 degrees C, and its activity increased threefold in the presence of 0.1-0.2 M NaCl or 0.2 M KCl. The Km values of recombinant DABAcT measured at the optimal pH and temperature in the presence of 0.2 M KCl were 460 and 36.6 microM for L-2,4-DAB and acetyl-CoA, respectively. The enzyme is specific for L-2,4-DAB and acetyl-CoA and is also active against propionyl-CoA (20%). Zn2+ and Cd2+ at 1 mM concentration completely inhibit the recombinant enzyme; 10 mM ATP inhibits 26% of the enzyme activity, whereas EDTA, o-phenanthroline, ADP, NAD(P), and NAD(P)H do not significantly effect the enzyme activity. The possible participation of DABAcT in regulation of ectoine biosynthesis in M. alcaliphilum 20Z is discussed. PMID- 16212545 TI - Structural-functional relationships between terminal deoxynucleotidyltransferase and 5'-triphosphates of nucleoside analogs. AB - Substrate properties of nucleoside 5'-triphosphate (NTP) analogs, namely, 5' triphosphates of L- and D-arabinonucleosides (D-FIAUTP, D-FMAUTP, and L-FMAUTP), D- and L-enantiomers of ddCTP analogs (D-ddCTP, L-ddCTP, D-FOddCTP, L-OddCTP, and L-SddCTP), and acyclic guanosine analogs (acyclovir and penciclovir) towards terminal deoxynucleotidyltransferase (TdT, EC 2.7.7.31) were studied. TdT can polymerize 5'-triphosphates of arabinonucleoside analogs (D-FIAUTP and D-FMAUTP). In contrast, L-FMAUTP is not recognized by TdT as a substrate. Kinetic parameters of D- and L-enantiomers of ddCTP analogs and 5'-triphosphates of acyclic nucleosides were evaluated. It is shown that stereospecificity of dNTP analogs and structure of the furanose residue play crucial roles in the interaction with TdT:L-enantiomers are much less potent as substrates compared to their D counterparts. 5'-Triphosphates of acyclovir (ACVTP) and penciclovir (PCVTP) are about two orders of magnitude less effective as substrates than nucleosides bearing furanose residues, with PCVTP being a better substrate than ACVTP. It can be assumed that the hydroxyl group of PCVTP mimics the 3'-hydroxyl group of the ribose residue and plays an important role in the interaction with TdT. PMID- 16212547 TI - Inhibition of family II pyrophosphatases by analogs of pyrophosphate and phosphate. AB - Imidodiphosphate (the pyrophosphate analog containing a nitrogen atom in the bridge position instead of oxygen) is a potent inhibitor of family II pyrophosphatases from Streptococcus mutans and Streptococcus gordonii (inhibition constant Ki approximately 10 microM), which is slowly hydrolyzed by these enzymes with a catalytic constant of approximately 1 min(-1). Diphosphonates with different substituents at the bridge carbon atom are much less effective (Ki = 1 6 mM). The value of Ki for sulfate (a phosphate analog) is only 12 mM. The inhibitory effect of the pyrophosphate analogs exhibits only a weak dependence on the nature of the metal ion (Mn, Mg, or Co) bound in the active site. PMID- 16212546 TI - Specific proteolysis of neuronal protein GAP-43 by calpain: characterization, regulation, and physiological role. AB - The mechanism of specific proteolysis of the neuronal protein GAP-43 in axonal terminals has been investigated. In synaptic terminals in vivo and in synaptosomes in vitro GAP-43 is cleaved only at the single peptide bond formed by Ser41; this is within the main effector domain of GAP-43. Proteolysis at this site involves the cysteine calcium-dependent neutral protease calpain. The following experimental evidences support this conclusion: 1) calcium-dependent proteolysis of GAP-43 in synaptosomes is insensitive to selective inhibitor of micro-calpain (PD151746), but it is completely blocked by micro- and m-calpain inhibitor PD150606; 2) GAP-43 proteolysis in the calcium ionophore A23187-treated synaptosomes is activated by millimolar concentration of calcium ions; 3) the pattern of fragmentation of purified GAP-43 by m-calpain (but not by micro calpain) is identical to that observed in synaptic terminals in vivo. GAP-43 phosphorylated at Ser41 by protein kinase C (PKC) is resistant to the cleavage by calpain. In addition, calmodulin binding to GAP-43 decreases the rate of calpain mediated GAP-43 proteolysis. Our results indicate that m-calpain-mediated GAP-43 proteolysis regulated by PKC and calmodulin is of physiological relevance, particularly in axonal growth cone guidance. We suggest that the function of the N-terminal fragment of GAP-43 (residues 1-40) formed during cleavage by m-calpain consists in activation of neuronal heterotrimeric GTP-binding protein G(o); this results in growth cone turning in response to repulsive signals. PMID- 16212548 TI - Chaperone-like activity of Mycobacterium tuberculosis Hsp16.3 does not require its intact (native) structures. AB - Small heat shock proteins (sHsps) were found to exhibit efficient chaperone-like activities under stress conditions although their native structures are severely disturbed. Here, using an alternative approach (site-directed mutagenesis), we obtained two structurally and functionally distinct Mycobacterium tuberculosis Hsp16.3 single-site mutant proteins. The G59W mutant protein (with Gly59 substituted by Trp) is capable of exhibiting efficient chaperone-like activity even under non-stress conditions although its secondary, tertiary, and quaternary structures are very different from that of the wild type protein. By contrast, the G59A mutant protein (with Gly59 substituted by Ala) resembles with the wild type protein in structure and function. These observations suggest that the Gly59 of the Hsp16.3 protein is critical for its folding and assembly. In particular, we propose that the exhibition of chaperone-like activity for Hsp16.3 does not require its intact (native) structures but requires the disturbance of its native structures (i.e., the native structure-disturbed Hsp16.3 retains its chaperone like activity or even becomes more active). In addition, the behavior of such an active mutant protein (G59W) also strongly supports our previous suggestion that Hsp16.3 exhibits chaperone-like activity via oligomeric dissociation. PMID- 16212549 TI - Rapid identification of Quox-1 homeodomain DNA-binding sequence using SAAB. AB - Quox-1 is the only gene in the hox family whose expression occurs throughout the development of the central nervous system. Using the Quox-1 homeodomain produced in a bacterial expression system, we were able to identify DNA-binding targets of the Quox-1 protein from a library of randomly generated oligonucleotides by the selection and amplification binding (SAAB) technique. The results indicated that the Quox-1 protein recognizes a new consensus sequence, 5'-CAATC-3', which has not been reported for any other Hox family homeoprotein. In addition, electromobility shift assay further confirmed that the Quox-1 homeoprotein preferentially binds to the 5'-CAATC-3' sequence, but not to the binding sites for other Hox class homeoprotein (TAAT) or NKX class homeoprotein (CAAG). Based on mutation analyses of the DNA sequences, we found that the 5'-CAATC-3' core sequences are required for high affinity binding by the Quox-1 protein. Furthermore, mutation analyses of the Quox-1 homeodomain showed that one of the major determinants participating in recognition of a minor groove is the Gln6 and Thr7 in the N-terminal arm of the homeodomain. PMID- 16212550 TI - Effects of cystine and hydrogen peroxide on glutathione status and expression of antioxidant genes in Escherichia coli. AB - Cysteine or cystine was earlier shown to multiply enhance the toxic effect of hydrogen peroxide on Escherichia coli cells. In the present work, the treatment of E. coli with H2O2 in the presence of cystine increased fivefold the level of extracellular oxidized glutathione (GSSG(out)) and decreased fivefold the GSH/GSSG(out) ratio (from 16.8 to 3.6). The same treatment of cells with deficiency in glutathione oxidoreductase (GOR) resulted in even more severe oxidation of GSH(out), so that the level of oxidized glutathione exceeded that of reduced glutathione and the GSH/GSSG(out) ratio decreased to 0.4. Addition of cystine to the GOR deficient cells resulted in significant oxidation of extracellular glutathione even in the absence of oxidant and in tenfold increase in intracellular oxidized glutathione along with a decrease in the GSH/GSSG(out) ratio from 282 to 26. However, in the cytoplasm of wild type cells, the level of oxidized glutathione (GSSG(in)) was changed insignificantly and the GSH/GSSG(in) ratio increased by 26% (from 330 to 415). Data on glutathione status and cystine reduction in the E. coli gsh and gor mutants suggested that exogenous cystine at first should be reduced with extracellular GSH outside the cells and then imported into them. The high toxicity of H2O2 in the presence of cystine resulted in disorders of membrane functions and inhibition of the expression of genes including those responsible for neutralization of oxidants and DNA repair. PMID- 16212551 TI - Characteristics of thiol:protein disulfide oxidoreductase from wheat (Triticum aestivum L.) grain. AB - Biochemical properties of a homogenous preparation of thiol:protein disulfide oxidoreductase (TPDO, EC 1.8.4.2) isolated for the first time from mature wheat (Triticum aestivum L.) grain were studied. According to polyacrylamide gel electrophoresis data, the molecular weight of TPDO is around 167 kD, the enzyme consisting of two subunits of 77 and 73 kD, which differentiates TPDO from known enzymes of SH/SS-metabolism of wheat caryopses. In substrate specificity and enzymatic characteristics (pH and temperature optima) TPDO is similar to analogous enzymes of animal tissues. Inhibition of disulfide reductase activity by alkylating agents and heavy metal ions suggests the participation of active center SH-groups in the catalytic act and classes the enzyme as a member of the thioredoxin superfamily. The SS-reductase reduces aggregating capacity of acetic acid-soluble fraction of wheat storage proteins. The proposed physiological role of TPDO is participation in creation and regulation of SH/SS-status of wheat endosperm proteins and formation of the rheological properties of gluten. PMID- 16212552 TI - Affinity modification of the restriction endonuclease SsoII by 2'-aldehyde containing double stranded DNAs. AB - Properties of 2'-aldehyde-containing double stranded DNAs (dsDNAs) have been studied for the first time as substrate analogs of the restriction endonuclease SsoII. These reactive oligonucleotides were successfully cross-linked to the restriction endonuclease SsoII by reductive amination, and conditions for DNA protein conjugate trypsinolysis followed by the oligonucleotide-peptide conjugate purification were optimized. Use of MALDI-TOF mass spectrometry revealed that covalent linkage forms between the sugar moiety of the central pyrimidine nucleoside of the SsoII recognition site and Lys173 of the enzyme. The latter is probably involved in initial steps of enzyme-substrate recognition during dsDNA readout. PMID- 16212553 TI - Inhibition of Naja naja venom hyaluronidase by plant-derived bioactive components and polysaccharides. AB - The inhibitory effect of several bioactive compounds on the activity of hyaluronidase enzyme purified from Naja naja venom was investigated in vitro. Compounds were found to inhibit the hyaluronidase activity dose dependently. Among glycosaminoglycans, heparin, heparan sulfate, and dermatan sulfate showed maximum inhibition compared to chondroitin sulfates. Different molecular forms of chitosan inhibit the enzyme, and inhibition appears to depend on the chain length. In addition, plant-derived bioactive compounds also inhibited the activity of hyaluronidase dose dependently. Among those tested, aristolochic acid, indomethacin, quercetin, curcumin, tannic acid, and flavone exhibited inhibition, with aristolochic acid and quercetin completely inhibiting the enzyme activity. It is concluded that the inhibitors of hyaluronidase could be used as potent first aid agents in snakebite therapy. Furthermore, these inhibitors not only reduce the local tissue damage but also retard the easy diffusion of systemic toxins and hence increase survival time. PMID- 16212554 TI - Herpes simplex virus type 1-encoded glycoprotein C contributes to direct coagulation factor X-virus binding. AB - The HSV1 (herpes simplex virus type 1) surface has been shown recently to initiate blood coagulation by FVIIa (activated Factor VII)-dependent proteolytic activation of FX (Factor X). At least two types of direct FX-HSV1 interactions were suggested by observing that host cell-encoded tissue factor and virus encoded gC (glycoprotein C) independently enhance FVIIa function on the virus. Using differential sedimentation to separate bound from free 125I-ligand, we report in the present study that, in the presence of Ca2+, FX binds directly to purified wild-type HSV1 with an apparent dissociation constant (K(d)) of 1.5+/ 0.4 muM and 206+/-24 sites per virus at saturation. The number of FX-binding sites on gC-deficient virus was reduced to 43+/-5, and the remaining binding had a lower K(d) (0.7+/-0.2 microM), demonstrating an involvement of gC. Engineering gC back into the deficient strain or addition of a truncated soluble recombinant form of gC (sgC), increased the K(d) and the number of binding sites. Consistent with a gC/FX stoichiometry of approximately 1:1, 121+/-6 125I-sgC molecules were found to bind per wild-type HSV1. In the absence of Ca2+, the number of FX binding sites on the wild-type virus was similar to the gC-deficient strain in the presence of Ca2+. Furthermore, in the absence of Ca2+, direct sgC binding to HSV1 was insignificant, although sgC was observed to inhibit the FX-virus association, suggesting a Ca2+-independent solution-phase FX-sgC interaction. Cumulatively, these data demonstrate that gC constitutes one type of direct FX HSV1 interaction, possibly providing a molecular basis for clinical correlations between recurrent infection and vascular pathology. PMID- 16212555 TI - Identification, cloning and characterization of a plasma membrane zinc efflux transporter, TrZnT-1, from fugu pufferfish (Takifugu rubripes). AB - An orthologue of the mammalian ZnT-1 (zinc transporter-1) gene was cloned from the intestine of the torafugu pufferfish (Takifugu rubripes), demonstrating that this gene predates the evolution of land-living vertebrates. TrZnT-1 (T. rubripes ZnT-1) shares overall topology with other members of the ZnT-1 family of zinc transporters, with six TMs (transmembrane domains) including a large histidine rich intracellular loop between TM IV and V and intracellular C- and N-termini. Expression of TrZnT-1 in a metallothionein acquiescent cell line suggested that this protein reduces intracellular Zn2+ levels. Manipulation of the transporting media showed that several externally applied hydrominerals had no effect on TrZnT 1 activity. However, addition of N-ethylmaleimide increased TrZnT-1-mediated transport, possibly by increasing intracellular free Zn2+ levels by Zn2+ release from carrier proteins. Generation of a specific antibody and subsequent immunocytochemistry on fixed cells overexpressing TrZnT-1 indicated that the protein is localized to the plasma membrane in these cells. The genomic organization of TrZnT-1 is the same as that in mammals with two exons. The upstream regulatory region of the TrZnT-1 gene contains several putative cis acting elements, including metal-response elements and an Sp1 site. Analysis of the DNA contigs surrounding the TrZnT-1 gene reveal limited synteny between corresponding regions in the rat, mouse and human; however, this was very low, with only two syntenic genes, ZnT-1 and NEK2 (never in mitosis gene A-related kinase). PMID- 16212559 TI - Editorial. PMID- 16212557 TI - Regulation of mitosis by poly(ADP-ribosyl)ation. AB - The spindle is a dynamic, microtubule-based structure responsible for chromosome segregation during cell division. Spindles in mammalian cells contain several thousand microtubules that are arranged into highly symmetric bipolar arrays by the actions of numerous microtubule-associated motor and non-motor proteins. In addition to these protein constituents, recent work has demonstrated that poly(ADP-ribose) is a key spindle component. Of the multitude of poly(ADP-ribose) polymerase proteins encoded in the genome, tankyrase 1 appears to be the primary enzyme responsible for building poly(ADP-ribose) in spindles during mitosis. In this issue of the Biochemical Journal, Susan Smith and co-workers show that the primary target of tankyrase 1 in dividing cells is NuMA (nuclear mitotic apparatus protein), a protein that cross-links microtubule ends at spindle poles. The impact of poly(ADP-ribosyl)ation on the biochemical function of NuMA remains murky at this time, but these new results represent the first step to clearing the view as to how poly(ADP-ribosyl)ation regulates cell division. PMID- 16212560 TI - Frequency and characteristics of side effects associated with antidepressant drugs. AB - Depression is among the most common of chronic health problems. WHO report predicts that depression will be the leading cause of disability in the industrial world by the year 2020. To be successful, treatment for the patients suffering from depression must be continued until complete recovery, but most patients do not stay on their antidepressant medication long enough. One of the most frequent reasons for break down is appearance of unpleasant side effects. In this study we followed up dynamics of the characteristic side effects of antidepressant therapy, with the major goal to assess their frequency and characteristics. The sample was all female patients taking antidepressant drugs in the Department of Psychiatry of Clinical Centre of University in Sarajevo. The treatment with antidepressants was efficient in most of the patients. A major advantage of SSRI over TCA was less pronounced side effects. The most intensive side effects of TCA (amitriptyline) were dry mouth, tremor and tachycardia while the most frequent side effects included blurred vision, tachycardia, dry mouth, tremor and sedation. Side effects of SSRI (fluoxetine/fluvoxamine) were mild, and the most frequent were nausea, tachycardia, swelling, dry mouth. PMID- 16212558 TI - Biochemical characterization of androgen receptor-interacting protein 4. AB - ARIP4 [AR (androgen receptor)-interacting protein 4] is a member of the SNF2-like family of proteins. Its sequence similarity to known proteins is restricted to the centrally located SNF2 ATPase domain. ARIP4 is an active ATPase, and dsDNA (double-stranded DNA) and ssDNA (single-stranded DNA) enhance its catalytic activity. We show in the present study that ARIP4 interacts with AR and binds to DNA and mononucleosomes. The N-terminal region of ARIP4 mediates interaction with AR. Kinetic parameters of the ARIP4 ATPase are similar to those of BRG-1 and SNF2h, two members of the SNF2-like protein family, but the specific activity of ARIP4 protein purified to >90% homogeneity is approximately ten times lower, being 120 molecules of ATP hydrolysed by an ARIP4 molecule per min in contrast with approx. 1000 ATP molecules hydrolysed per min by ATP-dependent chromatin remodellers. Unlike other members of the SNF2 family, ARIP4 does not appear to form large protein complexes in vivo or remodel mononucleosomes in vitro. ARIP4 is covalently modified by sumoylation, and mutation of six potential SUMO (small ubiquitin-related modifier) attachment sites abolished the ability of ARIP4 to bind DNA, hydrolyse ATP and activate AR function. We conclude that, similar to its closest homologues in the SNF2-like protein family, ATRX (alpha-thalassemia, mental retardation, X-linked) and Rad54, ARIP4 does not seem to be a classical chromatin remodelling protein. PMID- 16212561 TI - Isolation and immunochemical characterization of human cystitis C. AB - Cystatin C is a natural inhibitor of the cysteine proteinases papain, and mammalian lysosomal cathepsins B, H, L and S. This protein is thought to serve an important physiological role as a local regulator of enzyme activity. The changes of levels of cystatin C in extracellular fluids have shown themselves having potential clinical importance. We have purified cystatin C from urine of patients with chronic renal failure by procedure using affinity chromatography on CM papain Sepharose, gel filtration on Sephacryl S-200, and ion exchange chromatography on CM-cellulose. After isolation we obtained three inhibitory peaks (pI's from 7.8 to 9.2) which represent isoforms of the same protein. These isoforms are immunologically identical and differ in N-terminal sequence of the molecule. The form with pI 9.2 represents the intact inhibitor form, whereas the form with pI 7.8 is shortened for 8 amino-acid residues at N-terminal end. Purified cystatin C pI 9.2 was used for immunization of rabbits. Polyclonal antibodies, produced in rabbits, were isolated from rabbit sera by affinity chromatography on Protein A Sepharose. Enzyme immunoassay (ELISA) for cystatin C is developed on the basis of purified antibodies. Using ELISA test we determined amount of cystatin C in urine and serum samples of patients with chronic renal failure. The concentration of the inhibitor in the urine of these patients was approximately 100-fold more than in normal urine. In the serum from the same patients we found concentrations of cystatin C to be five times higher in comparison with the serum of healthy individuals. PMID- 16212556 TI - c-Cbl and Cbl-b ubiquitin ligases: substrate diversity and the negative regulation of signalling responses. AB - The activation of signalling pathways by ligand engagement with transmembrane receptors is responsible for determining many aspects of cellular function and fate. While these outcomes are initially determined by the nature of the ligand and its receptor, it is also essential that intracellular enzymes, adaptor proteins and transcription factors are correctly assembled to convey the intended response. In recent years, it has become evident that proteins that regulate the amplitude and duration of these signalling responses are also critical in determining the function and fate of cells. Of these, the Cbl family of E3 ubiquitin ligases and adaptor proteins has emerged as key negative regulators of signals from many types of cell-surface receptors. The array of receptors and downstream signalling proteins that are regulated by Cbl proteins is diverse; however, in most cases, the receptors have a common link in that they either possess a tyrosine kinase domain or they form associations with cytoplasmic PTKs (protein tyrosine kinases). Thus Cbl proteins become involved in signalling responses at a time when PTKs are first activated and therefore provide an initial line of defence to ensure that signalling responses proceed at the desired intensity and duration. PMID- 16212562 TI - Frequency of the antipsychotic therapy acute side effects in the treatment of acute psychosis. AB - Antipsychotic drugs produce a wide spectrum of physiological actions. Some of these effects differ among the various classes of antipsychotics. This medications have indications in the treatment of acute psychotic disorders. The main goal of this investigation was to determine the incidence and prevalence of the neuroleptic therapy acute side effects. The reason for this epidemiological investigation performing was the lack of knowledge of the exact neuroleptic therapy side effects incidence. Qualitative study on this problem has not been performed yet. Antipsychotic therapy side effects prevalence rate according to the literature data is ranging from 24% to 74%. Different prevalence rate is a consequence of different antipsychotic drug usage, different drug administration method and different side effects identification. On account of all these facts, we put the hypothesis on the correlation between the antipsychotic therapy and occurred side effects. Our experiment included all patients hospitalised from December 31st 1999 to January 31st 2000 in Intensive Care Unit of Biological Psychiatry Department of Psychiatric Clinic in Sarajevo. All patients were divided in three groups according to the applied therapy. All of them met ICD-10 criteria for schizophrenia (F20-29). During our study the following examinations were performed: psychiatric interview, BRPS, scale of side effects, psychophysiological tests, general clinical impression, scale of appetite, carbon hydrate needs scale. Psychiatric and statistical evaluations were done as well. The evaluation of our examination is showing successful results in all groups of patients. The improvement of psychopathological symptoms was insignificant. Reported side effects were minimal with low incidence rate and relatively high prevalence rate. Statistical tests were calculated from the obtained data after what the null hypothesis was rejected. Consequently, an alternative hypothesis was confirmed and it indicated that the acute side effects incidence and prevalence were within the range of expectation. Intensity of the recorded side effects was moderate to mild. On the basis of the obtained data, it has been concluded that applied antipsychotic agents did not induce more psychophysiological function impairments in the treated patients. Psychophysiological functions remained in physiological range limits and their changes were not significant. Neuroleptic therapy side effects were minimal, meaning no toxic signs or therapy discontinuations were recorded. PMID- 16212563 TI - Assessment of chronic neuropsychological effects of mercury vapour poisoning in chloral-alkali plant workers. AB - A prospective case study was conducted in the Department of Occupational Medicine, Tuzla. The purpose of this study was to indicate negative effects from occupational exposure to mercury on behavioural and mental health, memory and psychomotor function that was tested in 46 chloral-alkali plant workers (mean age was 38. 8+/- 5. 7 years; mean age of occupational history 16. 5+/- 6. 0 years). Data on toxicological monitoring on atomic absorption spectrometer, and data on mental health were collected, psychiatric and other subjective symptoms, and behavioural, psychomotor and memory function tested. The data were compared to control group, 32 healthy non exposed workers. The study was designed to assess blood and urine mercury levels and length of occupational exposure and investigate its relationships to effects on the mental health. The mean air mercury levels were 0.23 mg/m3, the mean blood mercury concentrations was 3. 6 mg/ dl and the mean urine mercury concentrations were 151.7 +/- 180.4 mg/l. In 25 (53%) workers exposed to mercury vapour was identified Depression-Hypochondrias Syndrome (p trend < 0. 001) with higher scores for scales: Hysteria (p trend <0. 001), Schizoid and Psychoastenia (MMPI). All psychological parameters were in highly significantly correlations with mercury levels and length of occupational exposure. Pathological parameters were possible general identified if the concentration of blood mercury levels are >2. 9 mg/ dl, or urine mercury levels > 87 mg/l workers exposed to mercury vapour knew that toxic effects in body resulted in loosing some of intellectual abilities, and that people who handle chemicals had an increased health risk (ESW questionnaire). The occupational mercury exposed workers had introvert behaviour (EPQ). Aggressiveness was found in 71.7% workers. The cognitive disturbances: short-term memory loss, difficult to concentrate on tasks which require attention and thinking, were significantly differed compared to those of controls (p trend < 0. 001). In 24 (52%) exposed to mercury workers we have determined ego strength loss and regressive defensive mechanisms (LB). Handwriting disturbances-micrography we have identified in 27 (58.7%) workers. PMID- 16212564 TI - Assessment of health effects in workers at gasoline station. AB - The aim of this study was to made assessment of health effects in 37 workers exposed to gasoline, and its constituents at gasoline stations between 1985 and 1996. Thirty-seven persons who had been exposed to gasoline for more than five years were examined. The evaluation included a medical / occupational history, haematological and biochemical examination, a physical exam, standardized psychological tests, and ultrasound examination of kidneys and liver. The groups were identical in other common parameters including age, gender (all men), and level of education (P<0. 05). The data were compared to two control groups: 61 healthy non-exposed controls and 25 workers at gasoline stations exposed to organic lead for only nine months. Peripheral smear revealed basophilic stippling and reticulocytosis. We found in chronic exposed gasoline workers haematological disorders: mild leukocytosis (7 of 37), lymphocytosis (20 of 37), mild lymhocytopenia (3 of 37), and decrease of red blood cells count (11 of 37). Results indicated that they have suffered from liver disorders: lipoid degeneration of liver (14 of 37), chronic functional damages of liver (3 of 37), cirrhosis (1 of 37). Ultrasound examination indicated chronic kidney damages (8 of 37). These results significantly differed from those of controls (P< 0.05). In 13 out of 37 workers at gasoline stations exposed to gasoline for more than 5 years the symptom of depression and decreased reaction time and motor abilities were identified. The summary of diseases of workers exposed to organic lead and gasoline are discussed. PMID- 16212565 TI - Mitochondrial medicine - a key to solve pathophysiology of xxi century diseases. AB - Over the past 13 years mitochondrial defects have been involved in wide variety of degenerative diseases - Parkinson disease, Alzheimer dementia, arteriosclerosis, ageing and cancer. Mitochondria are believed to control apoptosis or programmed cell death. Disturbance in mitochondrial metabolism has also been implicated in many common diseases such as congestive hart failure, diabetes and migraine. Scientific investigations have showed complexities in mitochondrial genetics, but at the same time, pathophysiology of mitochondrial diseases is still enigma. Mitochondria and their DNAs are opening the era of "mitochondrial medicine". What we today call "a mitochondrial medicine" is only a part of the whole panorama of diseases based on disordered mitochondrial function. PMID- 16212566 TI - Mathematical methods for quantification and comparison of dissolution testing data. AB - In recent years, drug release/dissolution from solid dosage forms has been the subject of intense and profitable scientific developments. Whenever a new solid dosage form is developed or produced, it is necessary to ensure that drug dissolution occurs in an appropriate manner. The pharmaceutical industry and the registration authorities do focus, nowadays, on drug dissolution studies. The quantitative analysis of the values obtained in dissolution/release tests is easier when mathematical formulas that express the dissolution results as a function of some of the dosage forms characteristics are used. This work discusses the analysis of data obtained for dissolution profiles under different media pH conditions using mathematical methods of analysis described by Moore and Flanner. These authors have described difference factor (f1) and similarity factor (f2), which can be used to characterise drug dissolution/release profiles. In this work we have used these formulas for evaluation of dissolution profiles of the conventional tablets in different pH of dissolution medium (range of physiological variations). PMID- 16212567 TI - Frequency of rh phenotypes in relation to the outcome of pregnancy in the two groups of pregnant women. AB - Lethal and sublethal genetical factors, including Rh factor, represent endogenous risk factors of the pregnancy outcome. These factors are most frequently inherited in recessive way and they often lead to the negative outcome of pregnancies. They represent pregnancy (a prirori) risk of various degrees. Inheritance of Rh system blood groups is linked to chromosome 1 and it could be explained by two alternative theories; molecular Rh system genetics has not yet been completely explained. The first formal-genetic theory postulates three closely linked gene sites (loci C, D and E) while the second theory has a monogenic character (one locus with several allele genes). Data on 755 pregnancies, which were (for various reasons) estimated as increased risk pregnancies, were registered at Gynaecology Clinic, Clinical Centre of University of Sarajevo, during the period from 1989 to 1992. These data were collected from pregnant women who, according to the certain indications from their familiar and personal anamnesis, demanded genetic consultations. The result of investigation of the basic Rh system phenotype distribution shows no statistically significant difference between monitored pregnant women. This result is assumed as valid for both pregnant women and their partners. The same result is suggesting that the observed increased risk pregnancy samples do not significantly differ from the previously studied population samples. Therefore, it has been concluded that Rh factor is not closely related to the increased risk of individual pregnancy outcomes, that is, it does not have relevant influence on the observed reproduction parameters. This result is very interesting and deserves particular medical attention and further evaluation in the future, particularly considering known immunological phenomena resulting from relations between reproduction partners belonging to the basic Rh system phenotypes. PMID- 16212568 TI - Posttraumatic stress disorder (ptsd) and co-morbidity. AB - Posttraumatic Stress Disorder (PTSD) very often occurs accompanied with other psychiatric disorders such as: Alcohol and Drug abuse, Personality Disorder, General Anxiety Disorder, Obsessive Compulsive Disorder, Schizophrenia etc. Sometimes it might be a problem for clinicians to differ PTSD symptoms from symptoms of coexisting psychic disorders. The aim of this study was to present the most common PTSD coexisting psycho-disorders. This research was conducted during the period from April 1998 to October 1999. Participants were divided in two groups each containing 30 examinees. The first group consisted of 30 participants with symptoms of PTSD only while the second group included participants who suffered from both PTSD and other psychic disorders (co morbidity). Both groups were quite similar regarding participants gender and age. The scientific tools used in the research were: Standard Psychiatric Interview, Harvard Trauma Questionnaire (HTQ), Hamilton Anxiety Rating Scale, Hamilton Depression Rating Scale, and Drug and Alcohol Abuse Checklist. Our research results are indicating that PTSD symptoms are most common in middle-aged persons, regardless of their gender and age. We have found following coexisting psychic disorders: personality disorder 46.6% (from which 13.3% is permanent personality disorder after the traumatic experience); depression 29.9% (depression without psychotic symptoms 23.3% and depression with coexisting psychotic symptoms 6.6%); drug abuse 13.3; alcohol abuse 6.7% and dissociative (conversion) disorder 3.3%. The results of our work are suggesting that co-morbid psychic symptoms have significant regressive influence on PTSD course and prognosis. PMID- 16212569 TI - The new trends in theophylline therapy. AB - Sustained-release theophylline pellets formulation for once-daily evening administration significantly improved patients compliance and adjusted serum levels profile of the drug. The patients conversion from i.v. to p.o. therapy is one of the most critical steps in the treatment of asthma according to its chronopathophysiological character. In our study we have examined safety and efficiency of this conversion in twelve hospitalised asthmatic patients who were given the new sustained-release theophylline pellets formulation for once-daily evening administration. The lung function parameters (FEV1, VC, RV, and Rt) and serum theophylline concentrations were monitored. So, the values obtained for the last day of i.v. therapy and the fifth day of p.o. therapy were compared. We found that 75% of the patients had no change or improved lung function on the conversion. Our results indicate that this conversion from i.v. to p.o. theophylline therapy is safe and could be efficacious. Also, the maximum theophylline serum levels could safely be predicted by measuring only one serum concentration in p.o. therapy with sustained-release theophylline pellets formulation for once-daily evening administration. PMID- 16212570 TI - Transcranial Doppler sonography as diagnostic method. AB - Having in mind the fact that cerebrovascular disease (CVB) takes today in medicine, in spite of diagnostic and therapeutic modernisation, the third place of mortality causes in the world (behind cordial and malignant diseases, but in front of depressions), and the second place of invalidity cause (right after trauma) as well as the second place of dementia cause (after Alzheimer disease), it urges primary prophylaxis. Developing countries, but before all countries of East and Middle Europe, where is our country, are highly risked areas where CV disease has trend of incidence and total frequency increase. In the neighbouring Croatia today CV disease is at the first place of mortality causes. In the world today 5 million people annually suffer CV disease, in Europe about 700.000, but frequency of suffering on Balkan is about 5 prom. Age of CV disease effecting unfortunately moved towards young age, and today 49% of effected by CV disease are of 46 to 59 years of age. Early detection and treatment of risk factors (before hypertension, smoking, diabetes mellitus, hyperlipidaemia, stress and physical non-activities) are the first aspect of CV disease prophylaxis. Together with this aspect of primary prophylaxis is early detection of complications of mentioned risk factors on the walls of blood vessels, before all changing in sense of arteriosclerosis, with consequence of disorder of cerebral haemodynamics. With that objective - verification of circulator and total haemodynamic disorders, there is obvious disclose of non-invasive diagnostic methods, and one of them is Transcranial Doppler Sonography (TCD). TCD is method comfortable for patient, reliable and rather precise, dynamic, and can be repeated several times, without side effects and in comparison with others rather cheap. PMID- 16212571 TI - Osteonecrosis of the jaws in periodontal patients with a history of bisphosphonates treatment. AB - BACKGROUND/AIM: Osteonecrosis of the jaws is being increasingly reported in patients with bone metastasis from a variety of solid tumours and disseminated multiple myeloma receiving intra-venous bisphosphonates. The signs and symptoms that may occur before the appearance of clinical evident osteonecrosis include changes in the health of periodontal tissues, non-healing mucosal ulcers, loose teeth and unexplained soft-tissue infection. A series of nine periodontally involving patients showing osteonecrosis of the jaws that appeared following the intra-venous use of bisphosphonates is reported. MATERIAL AND METHODS: Nine consecutive patients with osteonecrosis of the jaws were prospectically studied. Patients' past medical histories and the drugs that they had received for their malignant disease were systematically documented. Clinical, histopathological and radiographic features and proposal for treatment modalities of osteonecrosis are also reported. RESULTS: Of the nine patients (six women and three men) observed, all had osteonecrosis in the mandible; two had maxillary involvement as well. All nine patients had a history of extraction of periodontally hopeless teeth preceding the onset of osteonecrosis. In two patients, the lesions also appeared in edentulous areas spontaneously. All the patients had received intra-venous bisphosphonates as treatment for their disseminated haematological neoplasms or metastatic bone disease. The duration of bisphosphonate therapy at presentation ranged from 10 to 70 months (median: 33 months). CONCLUSIONS: Jaw osteonecrosis appears to be associated with the intra-venous use of bisphosphonates. Dental professionals should be aware of this potentially serious complication in periodontal patients receiving long-term treatment with bisphosphonates. PMID- 16212572 TI - Relationship between bleeding on probing and periodontal disease progression in community-dwelling older adults. AB - OBJECTIVE: The main objective of this study was to determine the relationship between bleeding on probing (BOP) and periodontal disease progression in community-dwelling older adults. METHODS: A 3-year longitudinal study was carried out in 229 non-smoking healthy older adults aged 70 years. Using pressure controlled periodontal probes, BOP, pocket depth and attachment level at 13,289 sites were measured annually. Periodontal disease progression was defined as an increase in attachment loss of >or=3 mm from the baseline to the final examination. The backward stepwise logistic regression analysis was performed to assess the relationship between the total number of sites with BOP in the four examinations and periodontal progression. RESULTS: Logistic regression analysis showed that the odds ratios of BOP frequency for periodontal disease progression ranged from 1.4 to 6.2 after controlling for pocket depth >or=4 mm at baseline, number of missing teeth, jaw type and tooth site. CONCLUSION: Increasing frequencies of bleeding might increase the probability of periodontal disease progression in community-dwelling older adults. PMID- 16212573 TI - Psychosocial factors as risk indicators of periodontitis. AB - OBJECTIVE: Cross-sectional investigation of associations between psychosocial and periodontal parameters. METHODS: One hundred and ten patients 18-76 years of age were examined clinically and 57 radiographically at the Department of Conservative Dentistry of the University Hospital of Heidelberg: probing depths (PDs) and attachment level (PAL-V) were obtained at six sites per tooth. Inter proximal bone loss was assessed in 57 patients on panoramic radiographs. Further chemical and general environmental sensitivity, somatization, and smoking status were assessed by several questionnaires. RESULTS: Significant correlations between severe bone loss and age (r=0.38, p=0.004) were observed. PAL-V and PD also correlated with age (r=0.45, p<0.001; r=0.37, p<0.001) and pack years (r=0.21, p=0.031; r=0.3, p=0.002). After adjustment for age, smoking, and sex a negative correlation between chemical odour sensitivity and bone loss, PD=4 mm as well as PAL-V=4 mm was observed. Further, a negative correlation was observed between gastrointestinal sensitivity and PAL-V=4 mm. Psychological stress correlated positively with bone loss. CONCLUSION: The results give evidence for associations of psychosocial factors and periodontal disease. Some environmental traits seem to be related to more favourable periodontal status. PMID- 16212574 TI - Specification and analysis of the effects of coping in research on the relationship between psychosocial stress and periodontal disease. PMID- 16212576 TI - Removal of Streptococcus mutans biofilm by bubbles. AB - BACKGROUND: Air bubbles had been shown to remove particles and bacteria from surfaces, but they had not yet been studied regarding the removal of mature biofilm from a surface. METHODS: Streptococcus mutans were grown as a biofilm on glass coverslips and were exposed to a fluid stream with or without bubbles. Three parameters (stream velocity, gas fraction, and bubble size) were varied in the bubble stream to determine which conditions best remove the biofilm. RESULTS: At low velocities bubbles enhance biofilm removal compared with the liquid alone. Stream conditions that were shown to be the most effective in removing biofilm were large bubbles at low gas fractions. CONCLUSIONS: These results suggest that flowing bubble streams may be a desirable feature to incorporate into oral hygiene products to remove accumulated biofilms such as dental plaque. PMID- 16212577 TI - The crossover design to evaluate the efficacy of plaque removal in tooth-brushing studies. AB - OBJECTIVES: To evaluate the crossover clinical trial design to assess plaque removal efficacy of the Sonicare Elite. MATERIAL AND METHODS: A single-cohort, 12 week, two-treatment, single-blind, crossover clinical trial recruited 45 subjects. Plaque was recorded using the modified Quigley and Hein index plaque index (PI). After screening, subjects used the toothbrush for 2 weeks and were reminded to abstain from tooth cleaning 12-18 h prior to appointments. At visit two, subjects were randomized to 2 or 2(1/2) min. brushing time. PIs was recorded pre- and post-brushing. Subjects brushed for the allocated time for a further 2 weeks. At visit 3, PIs were recorded pre- and post-brushing. Two weeks later, at visit 4, the subjects crossed over and the protocol was repeated. RESULTS: There was no evidence of a learning effect within each arm of the crossover. A significant period effect was detected; however, no significant treatment by period effect was found. 2(1/2) min. brushing removed more plaque at full mouth (p=0.037), smooth (p=0.012) and lingual (p=0.002) sites compared with 2 min. CONCLUSION: The crossover design is a valid model for assessing plaque removal efficacy in tooth-brushing studies where no carry-over effect is clinically plausible. PMID- 16212575 TI - Modulation of clinical expression of plaque-induced gingivitis: effects of personality traits, social support and stress. AB - BACKGROUND: Studies have shown an association between an acute stressful event and gingivitis. However, the possible effects of personality traits associated with stress resistance/susceptibility and current level of stress on the clinical expression of plaque-induced inflammation remain to be examined. The aim of this study was to characterize the subject-based clinical behaviour of the gingiva during experimental gingivitis in relation to personality profile, psychological stress and coping behaviour. METHODS: Ninety-six systemically and periodontally healthy subjects (mean age: 23.6+/-1.7 years), 46 males and 50 females, non smokers, participated in a randomized, split-mouth, localized experimental gingivitis trial. Prior to the trial, subjects were asked to complete self administered questionnaires evaluating personality traits (Hardiness scale and Courtauld Emotional Control Scale), subjective stress (Visual Analogue Scale Total Distress), social support (Multidimensional Scale of Perceived Social Support, MSPSS) and life events (Life Experiences Survey (LES)). The influence of psychosocial factors was investigated in the overall population as well as in two sub-populations with different inflammatory response to plaque accumulation. RESULTS: No significant relationships were found between gingival inflammation variables and psychological measures. No significant differences were detected between subjects with different susceptibilties to plaque-associated gingivitis for any considered psychological variable. A significant association between plaque variables and LES (negative) or MSPSS (positive) was found; however, the variance explained by the model was low. CONCLUSIONS: Differences in the current level of stress and psychosocial variables indicative of stress susceptibility do not account for variability in plaque accumulation and gingival inflammation during experimental gingivitis in young adults. PMID- 16212579 TI - Effect of the menstrual cycle on pain experience associated with periodontal therapy: randomized, pilot study. AB - OBJECTIVES: The aim of this pilot cross-over study was to compare preliminarily the pain perception of female patients undergoing periodontal debridement during menstrual or pre-menstrual phases (peri-menstrual period) with that observed during mid-menstrual phase (post-menstrual period). MATERIALS AND METHODS: Twenty women with moderate-to-advanced chronic periodontitis and regular menstrual cycles were asked to complete Corah's Dental Anxiety Scale (DAS) during the first debridement visit. Patients were randomly assigned to receive their first debridement visit during either their peri-menstrual or post-menstrual period. Debridement was performed in bilateral quadrants of patients during the periods. Pain levels for each quadrant were assessed with a Visual Analogue Scale (VAS), after each debridement visit. RESULTS: There was no significant correlation of order of treatment in the intensity of perceived pain during the periods (p<0.05). The median VAS scores were 22.0 and 15.2 mm in the peri-menstrual and the post-menstrual period, respectively. Increase in pain perception among females during their peri-menstrual period was significantly greater than their post-menstrual period (p<0.05). CONCLUSION: No clinical conclusions can be drawn at this stage as this pilot study did not have a sufficiently broad population to generalize these observations to all female periodontal patients. Providing clinicians with information about patients' menstrual cycle during debridement can alter the pain experience. PMID- 16212578 TI - Long-term treatment with sub-antimicrobial dose doxycycline has no antibacterial effect on intestinal flora. AB - AIM: The purpose of this study was to determine if a 9-month regimen of sub antimicrobial doxycycline (20 mg, bid) had an effect on either the intestinal or the vaginal microflora. MATERIAL AND METHODS: A total of 69 periodontally diseased subjects were randomized to receive drug or placebo control for a 9 month period. Stool specimens and vaginal swabs were collected at baseline and after 3 and 9 months of therapy. Samples were examined for total anaerobic counts, opportunistic pathogens, and doxycycline-resistant (>or=4 microg/ml) bacteria. All isolates that survived sub-culture were identified and their susceptibilities determined to six antibiotics. Analyses were performed to determine if treatment differences were present. RESULTS: The only statistically significant differences (p<0.05) between the two treatment groups occurred in the doxycycline-resistant counts at the baseline sample period for the faecal samples. This imbalance was before treatment initiation and the administration of the study drug. No between-treatment differences were detected at either the 3- or 9-month sample period either in the predominant bacterial taxa present or in their antibiotic susceptibilities. CONCLUSIONS: There was no evidence that sub antimicrobial doxycycline treatment exerted an effect on the composition or doxycycline resistance level of either the faecal or the vaginal microflora. PMID- 16212580 TI - Host response to microbial challenge following resective/non-resective periodontal therapy. AB - BACKGROUND: The host response to microbial challenge depends on the recruitment of homing leucocytes and may be related to the experience to infectious insults over years. PURPOSE: The aim of this study was to investigate the soft tissue reactions to de novo plaque formation at sites treated with either open flap debridement or with the use of resective means during periodontal therapy. MATERIAL AND METHODS: Fifteen patients, who had been treated for periodontal disease (severe generalized chronic periodontitis), participated in the study. Surgical therapy was performed using either gingivectomy (GV) or open flap debridement (OFD) procedures in a split mouth design. After 6 months of healing (day 0), two gingival biopsies were obtained, one from the GV- and one from the OFD-treated sites. The experimental gingivitis model was applied and plaque accumulation was allowed for 3 weeks. New biopsies were obtained from the remaining quadrants on day 21 of plaque formation. The biopsies were snap frozen and prepared for immunohistochemical analysis. RESULTS: Following 3 weeks of plaque accumulation, the size of the lesion in OFD sites was more than twice as large than that in GV sites (0.42 versus 0.19 mm2). In the GV units, the lesion was characterized by almost similar proportions of T cells (CD3+, 6.0%) and B cells (CD19+, 6.6%), while the ICT in OFD sites was dominated by B cells (13.8%). During the 3-week period of plaque formation the increase in cell densities of T and B cells was three times larger in OFD than in GV sites. The proportion of ELAM-1 (CD62+ cells) decreased in GV (-0.4%) and increased in OFD (0.9%) sites. CONCLUSIONS: The host response that occurred in the gingival sites treated with OFD was more pronounced than the reaction that under similar experimental conditions took place in the regenerated gingiva at sites treated by resective means. PMID- 16212581 TI - Coronally advanced flap with or without enamel matrix derivative for root coverage: a 2-year study. AB - AIM: The purpose of this study was to assess the ability of enamel matrix derivative (EMD) to improve root coverage with a coronally advanced flap (CAF) during a 2-year follow-up. METHODS: Fifteen patients each with two single and similar bilateral Miller Class I or II gingival recessions (30 recessions) were selected. Each recession was randomly assigned to the test group (CAF+EMD) or the control group (CAF only). Clinical parameters recorded at baseline and at 6, 12 and 24 months were recession depth (R), recession width (WR), probing depth (PD), clinical attachment level (CAL) and keratinized tissue (KT). RESULTS: Reduction of R resulted in a significant CAL gain in both groups, whereas PD was not altered. In the test group, R decreased from 4.07 mm (SD+/-0.59) at baseline to 0.47 mm (SD+/-0.74) at 24 months, corresponding to a mean root coverage (MRC) of 90.67%, whereas in the control group R shrank from 4.13 mm (SD+/-0.74) at baseline to 0.60 mm (SD+/-0.83) at 24 months (MRC=86.67%). Complete root coverage was achieved at 24 months in 73.33% and 60% of the two groups. A significant KT increase was observed in both groups. CONCLUSIONS: Root coverage outcomes were similar in both groups and no statistically significant differences were found at all between them. Hence, the additional use of EMD to CAF is not justified for clinical benefits of root coverage, but as an attempt of achieving periodontal regeneration rather than repair. PMID- 16212583 TI - Surgical approaches to recipient vessels of the fore- and hindlimbs for microvascular free tissue transfer in dogs. AB - OBJECTIVE: To develop and evaluate surgical approaches to the arteries and veins of the fore- and hindlimbs for use as potential recipient vessels for free tissue transfer. STUDY DESIGN: Experimental anatomic study. SAMPLE POPULATION: Canine cadavers (11): 2 preserved and 9 fresh cadavers. METHODS: Fore- and hindlimbs from 1 preserved cadaver injected with a pigmented silicone/barium mixture, through the common carotid artery and external jugular vein, were cut in 1 cm cross-sections. Tissue sections were used to identify the location of vessels >1 mm that could be used as recipient vessels for free tissue transfer. The other preserved cadaver was used to develop surgical approaches to these vessels. Three surgeons evaluated the written descriptions and illustrations for these approaches using fresh cadavers. Modifications to the surgical approaches were made based on recommendations from these surgeons. RESULTS: Six approaches were developed to isolate forelimb recipient vessels: palmar access, distal medial antebrachial, mid-antebrachial, proximal antebrachial, distal humeral, and mid humeral vascular access. Twelve approaches were developed to isolate recipient vessels of the hindlimb: plantar access, dorsal tarsal, cranial distal tibial, craniomedial distal tibial, lateral distal tibial, medial distal tibial, medial femorotibial, lateral distal femoral, medial femoral, proximal medial femoral, groin, and proximal lateral femoral vascular access. CONCLUSIONS: Six forelimb and 12 hindlimb sites were identified for surgical access to recipient vessels (>1 mm diameter) suitable for use in free tissue transfer for wound reconstruction. CLINICAL RELEVANCE: For reconstruction of complex wounds of the extremities of dogs, surgeons should consider use of readily accessible recipient vessels that would allow for free tissue transfer to the fore- and hindlimbs. PMID- 16212584 TI - Synovial membrane microarthroscopy of the equine midcarpal joint. AB - OBJECTIVE: To evaluate the value of microarthroscopy in the equine midcarpal joint using the vital stains methylene blue, trypan blue, neutral red, and Janus green B to observe components of the synovial lamina propria, vascular architecture, and synoviocytes. STUDY DESIGN: Experimental. ANIMALS: Ten horses. METHODS: Microarthroscopy of left and right midcarpal joints was performed with and without vital staining of the synovium. Four vital stains (methylene blue, trypan blue, neutral red, and Janus green B) were evaluated, with each stain used in 5 joints. Synovial biopsy specimens were collected from the dorsomedial and dorsolateral aspects of the joint. RESULTS: All dyes were biocompatible. At x 60 without vital staining, synovial surface topography, vascular network, and translucency were observed. Intra-articular vital dyes improved evaluation of synovial surface topography. At x 150 with vital staining, individual synoviocytes were clearly identified with all dyes, except neutral red. Although methylene blue provided the best in vivo microscopic differentiation of the structure of the intima, trypan blue had superior retention in conventionally processed synovial biopsies. CONCLUSIONS: Methylene blue, trypan blue, neutral red, and Janus green B stains can be used safely for microarthroscopy. Good visualization of cells and vascular network can be obtained by microarthroscopy, and microarthroscopic evaluation of the synovium compares favorably with conventional histologic evaluation of biopsy specimens. CLINICAL RELEVANCE: Microarthroscopy may be beneficial in both research and clinical diagnosis of equine articular diseases. PMID- 16212585 TI - Arthroscopic verification of ultrasonographic diagnosis of meniscal pathology in dogs. AB - OBJECTIVE: To determine the clinical usefulness of ultrasonography for diagnosis of meniscal pathology in dogs. STUDY DESIGN: Double-blind prospective study. ANIMALS: Dogs (n=10) with lameness localized to the stifle. MATERIALS AND METHODS: Sonographic examination of each affected stifle was performed by 1 ultrasonographer unaware of specific historical and clinical data for the dog. Sonographic findings were recorded, but not reviewed until study completion. Arthroscopic examination of the affected stifle was performed within 48 hours of ultrasonography by 1 surgeon unaware of ultrasonographic findings. Arthroscopic findings were recorded, but not reviewed until study completion. Two investigators compared the ultrasonographic and arthroscopic findings at study completion to determine the sensitivity, specificity, positive predictive value, and negative predictive value. RESULTS: Two of the 10 dogs had bilateral stifle evaluation. Twenty-four lateral and medial menisci, with normal and abnormal findings, were examined. The sensitivity and specificity for ultrasonographic diagnosis were 90.0% and 92.9%, respectively; positive and negative predictive values were 90.0% and 92.9%, respectively. CONCLUSIONS: Ultrasonography is highly specific and sensitive for diagnosing bucket handle tears of the medial meniscus in dogs. CLINICAL RELEVANCE: Ultrasonographic evaluation of menisci in dogs is a noninvasive method for accurately and efficiently diagnosing pathology, determining the need for surgical intervention, and providing comprehensive information to clients. PMID- 16212586 TI - Canine sacroiliac luxation: anatomic study of dorsoventral articular surface angulation and safe corridor for placement of screws used for lag fixation. AB - OBJECTIVE: To define a safe corridor in the dorsoventral plane to facilitate placement of screws inserted in lag fashion within the sacral body for fixation of sacroiliac fracture-luxation injuries in dogs. STUDY DESIGN: Anatomic study. SAMPLE POPULATION: Cadaveric canine sacra. METHODS: Canine sacra (n=45) were used for a radiographic study to define a safe corridor in the dorsoventral plane for placement of screws inserted in lag fashion for fixation of sacroiliac luxation in the dog. The defined safe corridor allowed drilling to a depth of 65% of the sacral width to ensure screw purchase of > or =60%. Effects of positioning and measurement techniques were evaluated. RESULTS: Eighty-seven safe corridors were measured. The mean articular surface was 100+/-4.52 degrees from horizontal. Mean maximum, optimum, and minimum safe corridor drill angles were 111+/- 4.57 degrees, 100+/-4.70 degrees, and 89+/-5.17 degrees, respectively, from the articular surface. Predicted surgeon error of +/-4 degrees was used to define the safe corridor for use clinically. CONCLUSIONS: In 91% of sacra, a drill angle of 100+/-4 degrees would remain ventral to the vertebral canal. Twelve sacra (14%) were at risk of penetration of the pelvic canal. A drill angle of 97+/-4 degrees avoids penetration of the vertebral canal in all sacra measured but risks ventral exit from the body in 30% of sacra studied. CLINICAL RELEVANCE: A drill angle of 97 degrees from the articular surface is recommended for insertion of screws for lag fixation of canine sacroiliac luxation. PMID- 16212587 TI - The effect of tibial plateau leveling osteotomy position on cranial tibial subluxation: an in vitro study. AB - OBJECTIVES: To compare centered versus distal tibial plateau leveling osteotomy (TPLO) position on cranial tibial subluxation, postoperative tibial plateau angle (TPA), and tibial long axis shift (TLAS). STUDY DESIGN: In vitro biomechanical evaluation. ANIMALS: Six pairs of canine cadaveric hind limbs. METHODS: One limb of each pair was randomly assigned to the distal (TPLO-D) or centered (TPLO-C) osteotomy group. Cranial tibial subluxation (CTS) under load was quantified sequentially under 3 conditions: intact, after cranial cruciate ligament transection, and after TPLO; a corrected CTS value was also calculated. Postoperative TPA and TLAS were measured. Comparisons were made using 1-way repeated measures ANOVA with a Tukey's multiple comparison post hoc test for CTS, and a Wilcoxon's sign rank test for TPA and TLAS. Significance was set at P<.05. RESULTS: TPLO-C had a significantly lower mean CTS than TPLO-D (P<.01). Corrected CTS was also significantly lower in TPLO-C than in TPLO-D (P<.001). Postoperative TPA and TLAS were less in TPLO-C than in TPLO-D (P=.0312). CONCLUSION: Our results confirm that distal centering of the TPLO leads to craniodistal translation of the tibial plateau, TLAS, and a postoperative TPA that is greater than expected. This geometric effect has the biomechanical effect expected of inadequate tibial plateau leveling, namely incomplete neutralization of cranial tibial thrust. CLINICAL RELEVANCE: The centered osteotomy position is geometrically more precise, and biomechanically more effective than the distal position. PMID- 16212588 TI - Comparison of perioperative analgesic protocols for dogs undergoing tibial plateau leveling osteotomy. AB - OBJECTIVE: To compare effects of 3 commonly used perioperative analgesic protocols (epidural injection, intra-articular injection, and intravenous [IV] injection) for management of postoperative pain in dogs after tibial plateau leveling osteotomy (TPLO). STUDY DESIGN: Prospective, randomized clinical trial. ANIMALS: Fifty-six healthy dogs with naturally occurring cranial cruciate ligament rupture. METHODS: Dogs were premedicated with IV hydromorphone and acepromazine and were randomly assigned to receive either E (preoperative epidural injection with morphine and bupivacaine), IA (pre- and postoperative intra-articular injections of bupivacaine), or C (neither epidural morphine and bupivacaine, nor intra-articular bupivacaine). All dogs were administered hydromorphone (0.05 mg/kg IV) at extubation and as needed to maintain comfort postoperatively. Patients were observed and monitored continuously for 24 hours and discomfort was assessed using visual analog pain scores (VASs), multifactorial pain scores (MPSs), and response to a pressure nociceptive threshold (PNT) measuring device. Time to 1st dose and the total doses of hydromorphone required to achieve adequate comfort for each dog were recorded. RESULTS: No differences in measured indices of postoperative pain were observed between dogs of each treatment group; VAS (P=.190), MPS (P=.371), and PNT (P=.160). Time to 1st analgesic intervention was longer for Group E compared with Group C (P=.005) and longer for Group IA compared with Group C (P=.032). Although time to 1st intervention between Groups E and IA were longer for Group E, differences were not significant. To provide an adequate level of comfort, more analgesic interventions were administered to dogs in Group C compared with dogs in group E (P=.015). On average, more hydromorphone was administered to Group C compared with Group IA (P=.072) and to Group IA compared with Group E (P=.168), but statistical significance was not reached for these data. CONCLUSIONS: In this study population, significant differences were seen in time to 1st hydromorphone dose between Groups E and IA compared with Group C. As well, more supplemental analgesia was administered to Group C compared with Group E to maintain the same level of postoperative comfort. Although differences between Groups E and IA tended to favor the epidural group, differences were minimal and not statistically significant. CLINICAL RELEVANCE: Our results suggest that regardless of analgesic protocol, measured indices of pain in dogs after TPLO can be minimized if dogs are continuously observed and appropriately supplemented with parenteral opioids. However, the frequency of postoperative opioid dosing can be minimized and may be a factor when contemplating supplementary use of epidural or intra-articular injections as part of a balanced analgesic approach. PMID- 16212589 TI - Short-term aseptic loosening of the femoral component in canine total hip replacement: effects of cementing technique on cement mantle grade. AB - OBJECTIVES: To evaluate the effects of different cementing techniques on radiographic cement mantle grade and short-term aseptic loosening of the femoral component in canine total hip replacement (THR). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Radiographs of 284 dogs that had THR. METHODS: Immediate postoperative radiographs of 284 dogs that had cemented THR were reviewed by 4 surgeons and 1 radiologist and assigned a cement mantle grade using a 4-grade classification system. Dog age and weight at surgery, cementing technique (1st, 2nd, 3rd generation), complications (type and timing), and follow up time were retrieved and analyzed. RESULTS: Mean cement mantle score for 2nd generation technique was significantly higher than 3rd generation technique and both were significantly higher than 1st generation technique (P<.001). Aseptic loosening was the 2nd most common complication observed with an incidence of 2.1%. Mean time from surgery to last follow-up examination was 122 days. No statistically significant difference in incidence of aseptic loosening was identified among different cement mantle grades or cementing techniques. CONCLUSIONS: Advanced cementing techniques resulted in better cement mantles based on grading of immediate postoperative radiographs, however grading did not predict short-term aseptic loosening. Cementing technique seemingly did not affect the incidence of short-term aseptic loosening of the femoral component for dogs in this study. CLINICAL RELEVANCE: Our study suggests that advances in cementing technique may result in improvements in the radiographic grade of cement mantles. With respect to aseptic loosening of the femoral component, our data only suggest that short-term (3 months-3 years) loosening cannot be predicted by immediate postoperative radiographic evaluation of cement mantle. PMID- 16212590 TI - Neodymium:yttrium aluminum garnet surgical laser versus bipolar electrocoagulation for laparoscopic ovariectomy in dogs. AB - OBJECTIVE: To compare use of neodymium:yttrium aluminum garnet (Nd:YAG) surgical laser and bipolar electrocoagulation (BEC) for laparoscopic ovariectomy (OVE) in dogs. STUDY DESIGN: Prospective clinical trial. ANIMALS: Female dogs (n=72). METHODS: Laparoscopic OVE by Nd:YAG laser (600 microm optical fiber, contact mode) in 36 dogs was compared with laparoscopic OVE by bipolar electro coagulating grasping forceps. Dogs were paired (laser, electrocoagulation) matched for breed, age, body weight, obesity, and number of heat cycles. Duration of predetermined surgery times and total surgical time were compared between groups. Occurrence of intra- and postoperative complications and their effect on surgical duration were evaluated. RESULTS: Laser surgery resulted in a higher incidence of intraoperative mesovarial bleeding (12 times; 9 dogs) compared with electrosurgery (4 times; 3 dogs). Use of laser caused a 2 minute delay for transection of the left ovary compared with electrosurgery. Postoperative complication rates and convalescence were similar for both groups. CONCLUSIONS: BEC reduced surgical time and intraoperative mesovarial bleeding compared with laser resection. CLINICAL RELEVANCE: Although the laser was effective for laparoscopic OVE, bipolar electrosurgical laparoscopic OVE remained the method of choice. PMID- 16212591 TI - Antebrachial fractures in four captive polar bears (Ursus maritimus). AB - OBJECTIVE: To identify common risk factors for antebrachial fractures of captive polar bears and to evaluate outcome after fracture repair. STUDY DESIGN: Retrospective study. ANIMALS: Four captive polar bears. METHODS: United States zoological collections were surveyed to determine the prevalence of fractures in captive polar bears. Medical records of captive polar bears that had antebrachial fractures were reviewed for signalment, history, physical and radiographic findings, fracture management, postoperative care, and outcome. Serum samples from healthy bears and bears with antebrachial fractures were assayed for 25 hydroxyvitamin D (25-OHD) concentrations. RESULTS: Nineteen fractures (12 polar bears) occurred from 1974 to 2002; 12 fractures involved the antebrachium. Management of 4 antebrachial fractures was reviewed; 3 were repaired by internal fixation and 1 by external coaptation. Fractures healed and bears were returned to exhibit on average 3 months postfracture. Of 11 serum samples assayed for 25 OHD concentrations, 6 were below normal, 1 was low normal and 4 were within normal reference intervals. The 7 bears with subnormal or low normal values were housed in 2 zoos. Subnormal vitamin D concentrations were identified in 2 of 3 bears with fractures. CONCLUSIONS: Fracture disease is not uncommon in captive polar bears. Additional research is necessary to explore the role of nutrition in polar bear fracture disease. CLINICAL RELEVANCE: Internal fixation of antebrachial fractures is feasible and reasonably well tolerated in captive polar bears. PMID- 16212592 TI - Teat tip reconstruction by supernumerary teat autotransplantation in cattle. AB - OBJECTIVE: Evaluation of use of a supernumerary teat as a pedicle graft to repair experimental teat tip damage in cattle. STUDY DESIGN: Experimental study. ANIMALS: Mature, non-gravid hybrid cows with > or =1 supernumerary teat. METHODS: The main teat and adjacent supernumerary teat were examined for patency by ultrasound with a 8 MHz linear array transducer. The tip of the main teat was resected at the junction of cistern and canal. The supernumerary teat was partially resected semi-circumferentially at the cranial aspect of the base to maintain vascular supply and transplanted by end-to-end anastomosis. The pedicle of the supernumerary teat graft was amputated in stages starting at 14 days, again at 16 days and separated at 18 days. Reconstructed teat patency was evaluated at intervals by ultrasonography and confirmed at 50 days by histopathology. Repeated measures ANOVA was used to compare the measurements of luminal diameter between days; significance was set at P<.05. RESULTS: All reconstructed teats were patent. Decreased luminal diameters were detected at 25, 32, 39, and 46 days by ultrasonography. On histopathology, granulation and epithelial (mucosal) proliferation contributed to luminal reduction. CONCLUSIONS: In this experimental model of an acute damage to a teat tip, supernumerary teat grafting was successfully used to restore teat anatomy. CLINICAL RELEVANCE: Supernumerary teat pedicle grafting may be considered as a salvage technique for reconstruction of a damaged bovine teat in cows that have an adjacent patent supernumerary teat. Function of the reconstructed teat in a lactating cow remains unknown. PMID- 16212593 TI - Comparison of three methods for arthrodesis of the distal intertarsal and tarsometatarsal joints in horses. AB - OBJECTIVE: To evaluate the effects of diode laser surgery (LS), surgical drilling (SD), and intraarticular sodium monoiodoacetate (MIA) as methods for fusing the distal intertarsal (DIT) and tarsometatarsal (TMT) joints in horses. STUDY DESIGN: Experimental study. ANIMALS: Adult horses (15) without radiographic signs of osteoarthritis (OA) of the DIT and TMT joints. METHODS: Group 1 (n=3) had LS performed bilaterally on DIT and TMT joints; 1 horse was evaluated for 1 week and 2 horses were evaluated for 2 weeks. Group 2 (n=6) had LS on DIT and TMT joints of 1 tarsus and MIA administration into the contralateral DIT and TMT joints and were evaluated for 6 months. Group 3 (n=6) had LS performed on DIT and TMT joints of 1 tarsus and SD of the contralateral DIT and TMT joints and were evaluated for 12 months. Postoperative comfort, lameness, radiography, microradiography, and histology scores were compared using repeated measures ANOVA, and paired or 2 sample t-tests; significance was set at P<.05. RESULTS: LS caused the least postoperative morbidity. In group 2, horses were less lame in 4 LS-treated limbs and 2 MIA-treated limbs at 6 months when compared with the contralateral limb. In group 3, horses were less lame in 5 LS-treated limbs and 1 SD-treated limb at 6 and 12 months compared with the contralateral limb. On microradiography, 11 MIA joints and 2 LS joints had bone bridging the joint at 6 months whereas 8 SD joints and 5 LS joints had bone bridging at 12 months. Significantly more joint space was bridged by bone in MIA- (51.4%) and SD (46.2%)-treated joints compared with LS joints at 6 (30.6%) and 12 (28.5%) months, respectively (P<.05). CLINICAL RELEVANCE: SD and MIA resulted in more bone bridging of the distal 2 tarsal joints, than LS. However, LS seemingly caused less pain and discomfort to horses in the immediate postoperative period; horses were generally less lame in the LS limb. More laser energy may need to be applied to these joints to promote fusion; however, it may also have beneficial effects beyond fusion. Further research on horses with OA of the distal 2 tarsal joints is needed to determine whether LS can cause soundness without facilitating bony fusion. PMID- 16212594 TI - Laser fenestration of the mesial septum for treatment of guttural pouch chondroids in a pony. AB - OBJECTIVE: To describe removal of guttural pouch chondroids through a laser fenestrated mesial septum in a pony with an obstructed pharyngeal ostium of the left guttural pouch. STUDY DESIGN: Case report. ANIMALS: Pony. METHODS: Transendoscopic laser neodymium:yttrium aluminum garnet (Nd:YAG) fenestration of the mesial guttural pouch septum was made by contact method (bare fiber, 15-25 W, exposure time 4 seconds). Dissected tissue was removed by transendoscopic monopolar electrosurgery (loop electrode, cutting blade) and multiple chondroids of varying size were removed from the left guttural pouch through the fenestration. RESULTS: After removing all chondroids and repeated flushing of the left guttural pouch via the right side, inflammation of the guttural pouch membranes decreased remarkably. The pony was discharged with instructions for transendoscopic lavage once weekly and was reevaluated at 2 months. A permanent septal fenestration was observed and the pony was free of abnormal clinical signs and being used for carriage driving competitions. CONCLUSION: Nd:YAG laser fenestration of the mesial septum is an approach for access to guttural pouch chondroids. CLINICAL RELEVANCE: When guttural pouch empyema and/or chondroids is associated with adhesion of the pharyngeal ostium, laser fenestration of the mesial septum from the opposite guttural pouch should be considered as an alternative approach for access, rather than classic external surgical approaches. PMID- 16212595 TI - Experimental use of polyamide bands in combination with intramedullary pinning for repair of oblique femoral fractures in rabbits. AB - OBJECTIVE: To evaluate the use of polyamide bands, manufactured for securing electrical cables, for repair of oblique femoral fractures in rabbits. STUDY DESIGN: Experimental study. ANIMALS: Twelve nulliparous, 21-25-week-old, California female rabbits. METHODS: Rabbits were divided into 3 groups (n=4) and studied for 14, 28, or 56 days. A Z-shaped mid-diaphyseal femoral osteotomy was repaired with a 2.5 mm intramedullary pin and a polyamide 6.6 cerclage band. Healing was evaluated at intervals by physical examination, limb use, femoral radiographs, and callus histology. RESULTS: Rabbits had early limb use with good wound healing. From the 1st day, movement of the hip and stifle joints was satisfactory. Radiographically, fractures were healed at 28 days. Histologically, there was no foreign body reaction and bone healing was normal. CONCLUSION: Nylon cerclage band application was accomplished easily, maintained reduction, and resulted in good healing and limb use. CLINICAL RELEVANCE: Sterilized nylon bands, manufactured for electrical use, can be used for cerclage in rabbits. PMID- 16212596 TI - Ureteral papilla implantation as a technique for neoureterocystostomy in cats. AB - OBJECTIVE: To evaluate ureteral papilla implantation for neoureterocystostomy in cats. STUDY DESIGN: Experimental study. ANIMALS: Five domestic shorthair cats. METHODS: The ureteral papilla was resected and the ureter isolated to the level of the renal hilus. The ureter was reimplanted into the apex of the bladder and the contralateral kidney was removed. Serum creatinine concentrations were measured daily for 7 days, then every other day from days 7-14, then every 3rd day until day 35. Ultrasound examination (once during the first 4 days, then again on day 35) and intravenous pyelography (day 35) were performed to assess ureteral patency. Histologic examination of the anastomosis was performed on day 35. RESULTS: Serum creatinine concentration remained within reference range for all cats except for a transient (<24 hour) increase (2.2, 2.3, and 3.6 mg/dL respectively) in 3 cats. Ultrasound examination and intravenous pyelography revealed no evidence of ureteral obstruction. Histologic examination revealed complete mucosal epithelialization of the anastomosis, moderate inflammation localized to residual suture material, and no evidence of vascular compromise of the ureteral papilla. CONCLUSION: Implantation of the ureteral papilla is a viable technique for neoureterocystostomy in cats. CLINICAL RELEVANCE: Advantages of ureteral papilla neoureterocystostomy include the technical ease of suturing the ureteral papilla, reduced risk of ureteral obstruction because sutures are not placed directly within the ureteral lumen and a more secure anastomosis because a complete 2-layer closure is performed. PMID- 16212597 TI - Multiple rib fracture in a neonatal foal using a nylon strand suture repair technique. AB - OBJECTIVES: To report rib fracture repair using the Securos Cranial Cruciate Ligament Repair System (SCCLRS; Securos Veterinary Orthopedics, Charlton, MA) in a neonatal foal. STUDY DESIGN: Case report. ANIMALS: A 1-day-old Standardbred foal with fracture of left ribs 2-9. METHODS: Four days after admission the foal was anesthetized and rib fractures were repaired using open reduction and the SCCLRS. RESULTS: Rib fractures were successfully stabilized and the foal was discharged 7 days postoperatively without further complications. CONCLUSIONS: The SCCLRS provided a straightforward, effective method of rib fracture repair in neonatal foals. CLINICAL RELEVANCE: Effective rib fracture repair in neonatal foals can be achieved with the SCCLRS. PMID- 16212598 TI - Use of autogenous fascia lata graft for perineal herniorrhaphy in dogs. AB - OBJECTIVE: To evaluate the effectiveness of a herniorrhaphy technique, using an autogenous fascia lata graft (FLG) for perineal hernia (PH) repair in dogs. STUDY DESIGN: Prospective clinical study. ANIMALS: Twelve dogs with PH. METHOD: PHs were repaired with FLG harvested from the dog's ipsilateral thigh and sutured directly into the perineal defect. Correction of associated conditions, and castration were performed. Surgical time, pain, inflammation, pattern of defecation, lameness, hospitalization time, postoperative complications, and owner satisfaction were recorded. Histopathologic examination was performed in 1 dog euthanatized 10 months after repair. RESULTS: Hernia did not recur (mean follow-up, 5.8 months). Lameness was the most frequent minor complication, and was resolved within a few days. Transient rectal prolapse occurred in 2 dogs with bilateral PH. The mean (+/-SD) hospitalization was 1.8+/-0.9 days, and the surgical time was 76.5+/-9.8 minutes. Histopathologic examination in 1 dog revealed perfect integration of FLG into adjacent tissues without substantial tissue reaction. CONCLUSIONS: FLG reconstruction of PH is a simple, effective method of treatment. CLINICAL RELEVANCE: FLG can be used without major complications for primary repair of PH, as an augmentation procedure when the internal obturator muscle is thin or friable, or when herniation has recurred after another repair technique. PMID- 16212599 TI - Cellular efflux of auxin catalyzed by the Arabidopsis MDR/PGP transporter AtPGP1. AB - Directional transport of the phytohormone auxin is required for the establishment and maintenance of plant polarity, but the underlying molecular mechanisms have not been fully elucidated. Plant homologs of human multiple drug resistance/P glycoproteins (MDR/PGPs) have been implicated in auxin transport, as defects in MDR1 (AtPGP19) and AtPGP1 result in reductions of growth and auxin transport in Arabidopsis (atpgp1, atpgp19), maize (brachytic2) and sorghum (dwarf3). Here we examine the localization, activity, substrate specificity and inhibitor sensitivity of AtPGP1. AtPGP1 exhibits non-polar plasma membrane localization at the shoot and root apices, as well as polar localization above the root apex. Protoplasts from Arabidopsis pgp1 leaf mesophyll cells exhibit reduced efflux of natural and synthetic auxins with reduced sensitivity to auxin efflux inhibitors. Expression of AtPGP1 in yeast and in the standard mammalian expression system used to analyze human MDR-type proteins results in enhanced efflux of indole-3 acetic acid (IAA) and the synthetic auxin 1-naphthalene acetic acid (1-NAA), but not the inactive auxin 2-NAA. AtPGP1-mediated efflux is sensitive to auxin efflux and ABC transporter inhibitors. As is seen in planta, AtPGP1 also appears to mediate some efflux of IAA oxidative breakdown products associated with apical sites of high auxin accumulation. However, unlike what is seen in planta, some additional transport of the benzoic acid is observed in yeast and mammalian cells expressing AtPGP1, suggesting that other factors present in plant tissues confer enhanced auxin specificity to PGP-mediated transport. PMID- 16212600 TI - Nod factors and a diffusible factor from arbuscular mycorrhizal fungi stimulate lateral root formation in Medicago truncatula via the DMI1/DMI2 signalling pathway. AB - Legumes form two different types of intracellular root symbioses, with fungi and bacteria, resulting in arbuscular mycorrhiza and nitrogen-fixing nodules, respectively. Rhizobial signalling molecules, called Nod factors, play a key role in establishing the rhizobium-legume association and genes have been identified in Medicago truncatula that control a Nod factor signalling pathway leading to nodulation. Three of these genes, the so-called DMI1, DMI2 and DMI3 genes, are also required for formation of mycorrhiza, indicating that the symbiotic pathways activated by both the bacterial and the fungal symbionts share common steps. To analyse possible cross-talk between these pathways we have studied the effect of treatment with Nod factors on mycorrhization in M. truncatula. We show that Nod factors increase mycorrhizal colonization and stimulate lateral root formation. The stimulation of lateral root formation by Nod factors requires both the same structural features of Nod factors and the same plant genes (NFP, DMI1, DMI2, DMI3 and NSP1) that are required for other Nod factor-induced symbiotic responses such as early nodulin gene induction and cortical cell division. A diffusible factor from arbuscular mycorrhizal fungi was also found to stimulate lateral root formation, while three root pathogens did not have the same effect. Lateral root formation induced by fungal signal(s) was found to require the DMI1 and DMI2 genes, but not DMI3. The idea that this diffusible fungal factor might correspond to a previously hypothesized mycorrhizal signal, the 'Myc factor', is discussed. PMID- 16212601 TI - The Rpi-blb2 gene from Solanum bulbocastanum is an Mi-1 gene homolog conferring broad-spectrum late blight resistance in potato. AB - The necessity to develop potato and tomato crops that possess durable resistance against the oomycete pathogen Phytophthora infestans is increasing as more virulent, crop-specialized and pesticide resistant strains of the pathogen are rapidly emerging. Here, we describe the positional cloning of the Solanum bulbocastanum-derived Rpi-blb2 gene, which even when present in a potato background confers broad-spectrum late blight resistance. The Rpi-blb2 locus was initially mapped in several tetraploid backcross populations, derived from highly resistant complex interspecific hybrids designated ABPT (an acronym of the four Solanum species involved:S. acaule, S. bulbocastanum, S. phureja and S. tuberosum), to the same region on chromosome 6 as the Mi-1 gene from tomato, which confers resistance to nematodes, aphids and white flies. Due to suppression of recombination in the tetraploid material, fine mapping was carried out in a diploid intraspecific S. bulbocastanum F1 population. Bacterial artificial chromosome (BAC) libraries, generated from a diploid ABPT-derived clone and from the resistant S. bulbocastanum parent clone, were screened with markers linked to resistance in order to generate a physical map of the Rpi-blb2 locus. Molecular analyses of both ABPT- and S. bulbocastanum-derived BAC clones spanning the Rpi blb2 locus showed it to harbor at least 15 Mi-1 gene homologs (MiGHs). Of these, five were genetically determined to be candidates for Rpi-blb2. Complementation analyses showed that one ABPT- and one S. bulbocastanum-derived MiGH were able to complement the susceptible phenotype in both S. tuberosum and tomato. Sequence analyses of both genes showed them to be identical. The Rpi-blb2 protein shares 82% sequence identity to the Mi-1 protein. Significant expansion of the Rpi-blb2 locus compared to the Mi-1 locus indicates that intrachromosomal recombination or unequal crossing over has played an important role in the evolution of the Rpi blb2 locus. The contrasting evolutionary dynamics of the Rpi-blb2/Mi-1 loci in the two related genomes may reflect the opposite evolutionary potentials of the interacting pathogens. PMID- 16212602 TI - Genetic analysis of two Arabidopsis DNA polymerase epsilon subunits during early embryogenesis. AB - Accurate DNA replication is one of the most important events in the life of a cell. To perform this task, the cell utilizes several DNA polymerase complexes. We investigated the role of DNA polymerase epsilon during gametophyte and seed development using forward and reverse genetic approaches. In Arabidopsis, the catalytic subunit of this complex is encoded by two genes, AtPOL2a and AtPOL2b, whereas the second largest regulatory subunit AtDPB2 is present as a unique complete copy. Disruption of AtPOL2a or AtDPB2 resulted in a sporophytic embryo defective phenotype, whilst mutations in AtPOL2b produced no visible effects. Loss of AtDPB2 function resulted in a severe reduction in nuclear divisions, both in the embryo and in the endosperm. Mutations in AtPOL2a allowed several rounds of mitosis to proceed, often with aberrant planes of division. Moreover, AtDPB2 was not expressed during development of the female gametophyte, which requires three post-meiotic nuclear divisions. Since a consensus binding site for E2F transcription factors was identified in the promoter region of both genes, the promoter-reporter fusion technique was used to show that luciferase activity was increased at specific phases of the cell cycle in synchronized tobacco BY-2 cells. Our results support the idea that fertilization may utilize the mechanisms of cell cycle transcriptional regulation of genes to reactivate the divisions of the oosphere and central cell. PMID- 16212603 TI - The tobacco plastid accD gene is essential and is required for leaf development. AB - Angiosperm plastid genomes typically encode approximately 80 polypeptides, mainly specifying plastid-localized functions such as photosynthesis and gene expression. Plastid protein synthesis and expression of the plastid clpP1 gene are essential for development in tobacco, indicating the presence of one or more plastid genes whose influence extends beyond the plastid compartment. The plastid accD gene encodes the beta-carboxyl transferase subunit of acetyl-CoA carboxylase and is present in the plastids of most flowering plants, including non photosynthetic parasitic plants. We replaced the wild-type accD gene with an aadA disrupted mutant allele using homologous recombination. Persistent heteroplasmy in the presence of antibiotics indicated that the wild-type accD allele was essential. The phenotype of the accD knockout was revealed in plastid transformants grown in the absence of antibiotics. Leaves contained pale green sectors and lacked part or all of the leaf lamina due to arrested division or loss of cells. Abnormal structures were present in plastids found in mutant plants, indicating that accD might be required to maintain the plastid compartment. Loss of the plastid compartment would be expected to be lethal. These results provide genetic evidence showing the essential role of plastid ACCase in the pathway leading to the synthesis of products required for the extra plastidic processes needed for leaf development. PMID- 16212605 TI - A duplicated pair of Arabidopsis RING-finger E3 ligases contribute to the RPM1- and RPS2-mediated hypersensitive response. AB - The Arabidopsis RPM1 protein confers resistance to disease caused by Pseudomonas syringae strains delivering either the AvrRpm1 or AvrB type III effector proteins into host cells. We characterized two closely related RPM1-interacting proteins, RIN2 and RIN3. RIN2 and RIN3 encode RING-finger type ubiquitin ligases with six apparent transmembrane domains and an ubiquitin-binding CUE domain. RIN2 and RIN3 are orthologs of the mammalian autocrine motility factor receptor, a cytokine receptor localized in both plasma membrane caveolae and the endoplasmic reticulum. RIN2 is predominantly localized to the plasma membrane, as are RPM1 and RPS2. The C-terminal regions of RIN2 and RIN3, including the CUE domain, interact strongly with an RPM1 N-terminal fragment and weakly with a similar domain from the Arabidopsis RPS2 protein. RIN2 and RIN3 can dimerize through their C-terminal regions. The RING-finger domains of RIN2 and RIN3 encode ubiquitin ligases. Inoculation with P. syringae DC3000(avrRpm1) or P. syringae DC3000(avrRpt2) induces differential decreases of RIN2 mobility in SDS-PAGE and disappearance of the majority of RIN2. A rin2 rin3 double mutant expresses diminished RPM1- and RPS2-dependent hypersensitive response (HR), but no alteration of pathogen growth. Thus, the RIN2/RIN3 RING E3 ligases apparently act on a substrate that regulates RPM1- and RPS2-dependent HR. PMID- 16212604 TI - Regulation of plant defense responses in Arabidopsis by EDR2, a PH and START domain-containing protein. AB - We have identified an Arabidopsis mutant that displays enhanced disease resistance (edr2) to the biotrophic powdery mildew pathogen Erysiphe cichoracearum. Inhibition of fungal growth on edr2 mutant leaves occurred at a late stage of the infection process and coincided with formation of necrotic lesions approximately 5 days after inoculation. Double-mutant analysis revealed that edr2-mediated resistance is suppressed by mutations that inhibit salicylic acid (SA)-induced defense signaling, including npr1, pad4 and sid2, demonstrating that edr2-mediated disease resistance is dependent on SA. However, edr2 showed normal responses to the bacterial pathogen Pseudomonas syringae pv. tomato strain DC3000. EDR2 appears to be constitutively transcribed in all tissues and organs and encodes a novel protein, consisting of a putative pleckstrin homology (PH) domain and a steroidogenic acute regulatory protein-related lipid-transfer (START) domain, and contains an N-terminal mitochondrial targeting sequence. The PH and START domains are implicated in lipid binding, suggesting that EDR2 may provide a link between lipid signaling and activation of programmed cell death mediated by mitochondria. PMID- 16212606 TI - hca: an Arabidopsis mutant exhibiting unusual cambial activity and altered vascular patterning. AB - By screening a T-DNA population of Arabidopsis mutants for alterations in inflorescence stem vasculature, we have isolated a mutant with a dramatic increase in vascular tissue development, characterized by a continuous ring of xylem/phloem. This phenotype is the consequence of premature and numerous cambial cell divisions in both the fascicular and interfascicular regions that result in the loss of the alternate vascular bundle/fiber organization typically observed in Arabidopsis stems. The mutant was therefore designated high cambial activity (hca). The hca mutation also resulted in pleiotropic effects including stunting and a delay in developmental events such as flowering and senescence. The physiological characterization of hca seedlings in vitro revealed an altered auxin and cytokinin response and, most strikingly, an enhanced sensitivity to cytokinin. These results were substantiated by comparative microarray analysis between hca and wild-type plants. The genetic analysis of hca indicated that the mutant phenotype was not tagged by the T-DNA and that the hca mutation segregated as a single recessive locus, mapping to the long arm of chromosome 4. We propose that hca is involved in mechanisms controlling the arrangement of vascular bundles throughout the plant by regulating the auxin-cytokinin sensitivity of vascular cambial cells. Thus, the hca mutant is a useful model for examining the genetic and hormonal control of cambial growth and differentiation. PMID- 16212607 TI - Tobacco BY-2 cells expressing fission yeast cdc25 bypass a G2/M block on the cell cycle. AB - The mitotic inducer gene from Schizosaccharomyces pombe, Spcdc25, was used as a tool to investigate regulation of G2/M in higher plants using the BY-2 (Nicotiana tabacum) cell line as a model. Spcdc25-expressing BY-2 cells exhibited a reduced mitotic cell size through a shortening of the G2 phase. The cells often formed isodiametric double files both in BY-2 cells and in cell suspensions derived from 35S::Spcdc25 tobacco plants. In Spcdc25-expressing cells, the tobacco cyclin dependent kinase, NtCDKB1, showed high activity in early S phase, S/G2 and early M phase, whereas in empty vector cells CDKB1 activity was transiently high in early S phase but thereafter remained lower. Spcdc25-expressing cells also bypassed a block on G2/M imposed by the cytokinin biosynthetic inhibitor lovastatin (LVS). Surprisingly, cytokinins were at remarkably low levels in Spcdc25-expressing cells compared with the empty vector, explaining why these cells retained mitotic competence despite the presence of LVS. In conclusion, synchronised Spcdc25-expressing BY-2 cells divided prematurely at a small cell size, and they exhibited premature, but sustained, CDKB1 activity even though endogenous cytokinins were virtually undetectable. PMID- 16212608 TI - ELF4 is a phytochrome-regulated component of a negative-feedback loop involving the central oscillator components CCA1 and LHY. AB - Evidence has been presented that a negative transcriptional feedback loop formed by the genes CIRCADIAN CLOCK ASSOCIATED (CCA1), LATE ELONGATED HYPOCOTYL (LHY) and TIMING OF CAB (TOC1) constitutes the core of the central oscillator of the circadian clock in Arabidopsis. Here we show that these genes are expressed at constant, basal levels in dark-grown seedlings. Transfer to constant red light (Rc) rapidly induces a biphasic pattern of CCA1 and LHY expression, and a reciprocal TOC1 expression pattern over the first 24 h, consistent with initial induction of this synchronous oscillation by the light signal. We have used this assay with wild-type and mutant seedlings to examine the role of these oscillator components, and to determine the function of ELF3 and ELF4 in their light regulated expression. The data show that whereas TOC1 is necessary for light induced CCA1/LHY expression, the combined absence of CCA1 and LHY has little effect on the pattern of light-induced TOC1 expression, indicating that the negative regulatory arm of the proposed oscillator is not fully functional during initial seedling de-etiolation. By contrast, ELF4 is necessary for light-induced expression of both CCA1 and LHY, and conversely, CCA1 and LHY act negatively on light-induced ELF4 expression. Together with the observation that the temporal light-induced expression profile of ELF4 is counter-phased to that of CCA1 and LHY and parallels that of TOC1, these data are consistent with a previously unrecognized negative-feedback loop formed by CCA1/LHY and ELF4 in a manner analogous to the proposed CCA1/LHY/TOC1 oscillator. ELF3 is also necessary for light-induced CCA1/LHY expression, but it is neither light-induced nor clock regulated during de-etiolation. Taken together, the data suggest (a) that ELF3, ELF4, and TOC1 all function in the primary, phytochrome-mediated light-input pathway to the circadian oscillator in Arabidopsis; and (b) that this oscillator consists of two or more interlocking transcriptional feedback loops that may be differentially operative during initial light induction and under steady-state circadian conditions in entrained green plants. PMID- 16212609 TI - Immediate-early and delayed cytokinin response genes of Arabidopsis thaliana identified by genome-wide expression profiling reveal novel cytokinin-sensitive processes and suggest cytokinin action through transcriptional cascades. AB - Cytokinins are hormones that regulate many developmental and physiological processes in plants. Recent work has revealed that the cytokinin signal is transduced by two-component systems to the nucleus where target genes are activated. Most of the rapid transcriptional responses are unknown. We measured immediate-early and delayed cytokinin responses through genome-wide expression profiling with the Affymetrix ATH1 full genome array (Affymetrix Inc., Santa Clara, CA, USA). Fifteen minutes after cytokinin treatment of 5-day-old Arabidopsis seedlings, 71 genes were upregulated and 11 genes were downregulated. Immediate-early cytokinin response genes include a high portion of transcriptional regulators, among them six transcription factors that had previously not been linked to cytokinin. Five plastid transcripts were rapidly regulated as well, indicating a rapid transfer of the signal to plastids or direct perception of the cytokinin signal by plastids. After 2 h of cytokinin treatment genes coding for transcriptional regulators, signaling proteins, developmental and hormonal regulators, primary and secondary metabolism, energy generation and stress reactions were over-represented. A significant number of the responding genes are known to regulate light (PHYA, PSK1, CIP8, PAT1, APRR), auxin (Aux/IAA), ethylene (ETR2, EIN3, ERFs/EREBPs), gibberellin (GAI, RGA1, GA20 oxidase), nitrate (NTR2, NIA) and sugar (STP1, SUS1) dependent processes, indicating intense crosstalk with environmental cues, other hormones and metabolites. Analysis of cytokinin-deficient 35S:AtCKX1 transgenic seedlings has revealed additional, long-lasting cytokinin-sensitive changes of transcript abundance. Comparative overlay-analysis with the software tool mapman identified previously unknown cytokinin-sensitive metabolic genes, for example in the metabolism of trehalose-6-phosphate. Taken together, we present a genome-wide view of changes in cytokinin-responsive transcript abundance of genes that might be functionally relevant for the many biological processes that are governed by cytokinins. PMID- 16212610 TI - Virus-induced gene silencing is an effective tool for assaying gene function in the basal eudicot species Papaver somniferum (opium poppy). AB - Virus-induced gene silencing (VIGS) is an attractive method for assaying gene function in species that are resistant to conventional genetic approaches. However, VIGS has been shown to be effective in only a few, closely related plant species. Tobacco rattle virus (TRV), a bipartite RNA virus, has a wide host range and so in principle could serve as an efficient vector for VIGS in a diverse array of plant species. Here we show that a vector based on TRV sequences is effective at silencing the endogenous phytoene desaturase (PapsPDS) gene in Papaver somniferum (opium poppy). We show that this vector does not compromise the growth or reproduction of poppy and the plants did not display viral symptoms. The silencing of PapsPDS resulted in a significant reduction in PapsPDS mRNA and a concomitant photobleached phenotype. The ability to rapidly assay gene function in P. somniferum will be valuable in manipulation of the opiate pathway in this pharmaceutically important species. We suggest that our vacuum infiltration method used to deliver TRV-based vectors into poppy is a promising approach for expanding VIGS to diverse angiosperm species in which traditional delivery methods fail to induce VIGS. Furthermore, these studies demonstrate the utility of TRV-VIGS for probing gene function in a basal eudicot species that is phylogenetically distant from model plant species. PMID- 16212611 TI - Sensitive detection and localization of hydroxyl radical production in cucumber roots and Arabidopsis seedlings by spin trapping electron paramagnetic resonance spectroscopy. AB - As reactive oxygen species are important for many fundamental biological processes in plants, specific and sensitive techniques for their detection in vivo are essential. In particular, the analysis of hydroxyl radical (OH*) formation in biological reactions has rarely been attempted. Here, it is shown that spin trapping electron paramagnetic resonance (EPR) spectroscopy allows the detection and quantitative estimation of OH* production in vivo in one single cucumber seedling root. It is possible to localize the OH* production site to the growth zone of the root by varying the position of the intact seedling inside the resonator cavity of the EPR spectrometer. Moreover, the demonstration of impaired OH* formation in the root of the Arabidopsis mutant rhd2 impaired in a superoxide producing Nicotimamide adenine dinucleotide phosphate (NADPH) oxidase has been accomplished. Spin trapping EPR provides a valuable tool for analyzing the production of OH*in vivo with high resolution in small tissue samples. PMID- 16212612 TI - Basal resistance against bacteria in Nicotiana benthamiana leaves is accompanied by reduced vascular staining and suppressed by multiple Pseudomonas syringae type III secretion system effector proteins. AB - Basal resistance in plants is induced by flagellin and several other common bacterial molecules and is implicated in the immunity of plants to most bacteria and other microbes. However, basal resistance can be suppressed by effector proteins that are injected by the type III secretion system (TTSS) of pathogens such as Pseudomonas syringae. This study demonstrates that basal resistance in the leaves of Nicotiana benthamiana is accompanied by reduced vascular flow into minor veins. Reduced vascular flow was assayed by feeding leaves, via freshly excised petioles, with 1% (weight in volume, w/v) neutral red (NR) and then observing differential staining of minor veins or altered levels of extractable dye in excised leaf samples. The reduced vascular staining was localized to tissues expressing basal resistance and was observable when resistance was induced by either the non-pathogen Pseudomonas fluorescens, a TTSS-deficient mutant of P. syringae pv. tabaci, or flg22 (a flagellin-derived peptide elicitor of basal resistance). Nicotiana benthamiana leaf areas expressing basal resistance no longer elicited the hypersensitive response when challenge inoculated with P. syringae pv. tomato DC3000. The reduced vascular staining effect was suppressed by wild-type P. syringae pv. tabaci and P. fluorescens heterologously expressing a P. syringae TTSS and AvrPto1(PtoJL1065). TTSS proficient P. fluorescens was used to test the ability of several P. syringae pv. tomato DC3000 effectors for their ability to suppress the basal resistance associated reduced vascular staining effect. AvrE(PtoDC3000), HopM1(PtoDC3000) (formerly known as HopPtoM), HopF2(PtoDC3000) (HopPtoF) and HopG1(PtoDC3000) (HopPtoG) suppressed basal resistance by this test, whereas HopC1(PtoDC3000) (HopPtoC) did not. In summary, basal resistance locally alters vascular function and the vascular dye uptake assay should be a useful tool for characterizing effectors that suppress basal resistance. PMID- 16212615 TI - Management of tuberculosis in the transplant recipient. PMID- 16212616 TI - A positive crossmatch and treatment with IvIg. PMID- 16212617 TI - Understanding the influence of ethnicity on renal allograft survival. PMID- 16212618 TI - Small-for-size syndrome after partial liver transplantation: definition, mechanisms of disease and clinical implications. AB - Widespread application of cadaveric split or living donor liver transplantation bears considerable potential to increase the pool of available organs and thus alleviate the problem of organ shortage. Although splitting of a cadaveric liver into two grafts for adult recipients can be performed successfully, sufficient function of undersized grafts is a major concern. To minimize the risk for living donors, transplant surgeons aim at procuring the least necessary liver volume, also leading to potentially small grafts. When small partial grafts are unable to meet the functional demands, the recipients can develop a so-called small-for size syndrome (SFSS). There is currently limited data on the pathogenesis of SFSS, with clinical studies mainly focusing on portal hyperperfusion. Additional aspects include graft-related factors such as functional and regenerative capacity, as well as recipient-related factors, such as overall health status and severity of cirrhosis. However, there is currently no consensus on the definition of SFSS. We propose a novel definition, based on simple clinical criteria, which divides the syndrome into either nonfunction or dysfunction of a small graft after the exclusion of other causes. This definition should ease comparability of future clinical trials, and thus improve understanding of the pathogenesis of SFSS. PMID- 16212619 TI - Humoral rejection of organ allografts. AB - In recent years, the deleterious clinical consequences of recipient de novo alloantibody production against HLA antigens from human organ allografts have been extensively investigated. In kidney transplantation, the identification of the complement C4d fragment in peritubular capillaries as a specific marker for humoral rejection has helped to define and characterize distinct clinical alloantibody-mediated syndromes. This knowledge is relevant for patient management as new therapeutic strategies to remove and control anti-donor antibody production, particularly in the setting of acute humoral rejection, have been reported. For recipients of nonrenal organ allografts such as heart transplant recipients, de novo anti-HLA alloantibody may also be important, although more studies are needed before clear guidelines can be proposed. PMID- 16212620 TI - More on parental living liver donation for children with fulminant hepatic failure: addressing concerns about competing interests, coercion, consent and balancing acts. AB - Parental living liver donation for children with fulminant hepatic failure raises complex ethical issues. According to a recent editorial in this journal, these include contradictory interests, the possibility of coercion and compromised consent and the need to balance the risks to the donor against the potential benefits for the recipient. Here I argue that in this setting, interests are often aligned rather than conflicted, that coercion of parental donors is rare, that consent may sometimes be valid even when it is not fully informed and that the correct balance to consider is the relative weights of risks and benefits for the donor. I conclude that living liver donation by parents of children with fulminant hepatic failure is consistent with societal norms of parental behavior, ethically acceptable and should be permitted regardless of the efficiency of the deceased donor organ recovery program. PMID- 16212622 TI - Composite islet-endothelial cell grafts: a novel approach to counteract innate immunity in islet transplantation. AB - An instant blood-mediated inflammatory reaction (IBMIR) is elicited when islets come in contact with blood after intraportal transplantation. In contrast, endothelial cells (EC) readily tolerate contact with blood. A conceivable strategy to overcome IBMIR would be to create composite islet-EC grafts. Human islets were co-cultured with primary human aortic endothelial cells (HAEC) for 2 7 days to obtain 50-90% coverage. HAEC-coated islets were exposed to ABO identical blood and analyzed with regard to clotting time, signs of inflammation and cell infiltration. Composite islet-HAEC graft survival was assessed after transplantation to athymic (nu/nu) nude mice. Exposed to blood, HAEC-coated islets induced less activation of coagulation and complement compared to control islets. Also, platelet and leukocyte consumption in blood was decreased. Clots with entrapped HAEC-coated islets showed less infiltration of CD11b+ cells. The extent of protection correlated to the level of HAEC coverage. Transplanted composite grafts stained positive for insulin and PECAM-1 demonstrating presence of both islets and HAEC within the islet graft 7 weeks after transplantation. Composite islet-HAEC grafts reduce all components of IBMIR. Refinement of the technique will allow introduction of composite islet-EC grafts in clinical islet transplantation, using autologous EC expanded in vitro and kept frozen until allogeneic islets become available for that specific recipient. PMID- 16212621 TI - Signaling T-cell survival and death by IL-2 and IL-15. AB - Interleukin 2 (IL-2) and interleukin 15 (IL-15) bind to common T-cell surface receptors comprised of unique alpha (IL-2R alpha or IL-15R alpha) and shared beta/gamma chain subunits. Ligation of this receptor by IL-2 can lead to apoptosis whereas IL-15 ligation favors cell survival. Our study examined intra cellular signaling events associated with IL-2- and IL-15-induced apoptosis and survival in human T cells. We found IL-2 and IL-15 could both induce apoptosis and survival; the outcome depended on cytokine concentration. No qualitative differences in Jak/Stat, Ras/MAPK or PI3K/AKT signaling were seen over a wide range of IL-2 and IL-15 concentrations. These findings suggest that, like T-cell receptor signaling, IL-2R beta/gamma chain signaling is regulated, or "tuned," by the concentration of cytokine. PMID- 16212623 TI - Endothelial cell calpain activity facilitates lymphocyte diapedesis. AB - Lymphocyte infiltration of tissue is a cardinal feature of solid-organ allograft rejection. Vascular endothelial cells (EC) participate in lymphocyte recruitment through the display of adhesion molecules and chemokines to promote leukocyte extravasation. Moreover, EC reorganize the cytoskeleton and cytoskeleton associated structures during leukocyte diapedesis. We examined the role of EC (Ca+2)i and the calcium-sensitive protease, calpain, during lymphocyte diapedesis through a human EC monolayer under physiologic shear stress in vitro. We observed that lymphocyte transendothelial migration (TEM) was inhibited by chelating EC cytosolic calcium, or depleting EC endoplasmic reticulum calcium stores by inhibition of the endoplasmic reticulum Ca ATPase. Further, inhibition of EC phospholiase C also decreased lymphocyte TEM. We determined that EC constitutively exhibit calpain activity, using fluorescence generation from a calpain substrate to report calpain activity in individual live cells. Moreover, EC adjacent to a transmigrating lymphocyte showed increased calpain activity. Further, lymphocyte TEM was inhibited by agents that block calpain activity. Inhibition of lymphocyte TEM occurs at the lumenal EC surface and correlates with impaired development of intercellular adhesion molecule 1 (ICAM-1)-rich docking structures by the EC. We conclude EC calcium and calpain activity facilitates lymphocyte TEM, and participates in the assembly of the docking structure. PMID- 16212624 TI - The sphingosine-1-phosphate receptor agonist FTY720 modulates dendritic cell trafficking in vivo. AB - The pro-drug FTY720 is undergoing phase III clinical trials for prevention of allograft rejection. After phosphorylation, FTY720 targets the G protein-coupled sphingosine-1-phosphate receptor 1 (S1PR1) on lymphocytes, thereby inhibiting their egress from lymphoid organs and their recirculation to inflammatory sites. Potential effects on dendritic cell (DC) trafficking have not been evaluated. Here, we demonstrate the expression of all five S1PR subtypes (S1PR1-5) by murine DCs. Administration of FTY720 to C57BL/10 mice markedly reduced circulating T and B lymphocytes within 24 h, but not blood-borne DCs, which were enhanced significantly for up to 96 h, while DCs in lymph nodes and spleen were reduced. Numbers of adoptively transferred, fluorochrome-labeled syngeneic or allogeneic DCs in blood were increased significantly in FTY720-treated animals, while donor derived DCs and allostimulatory activity for host naive T cells within the spleen were reduced. Administration of the selective S1PR1 agonist SEW2871 significantly enhanced circulating DC numbers. Flow analysis revealed that CD11b, CD31/PECAM-1, CD54/ICAM-1 and CCR7 expression on blood-borne DCs was downregulated following FTY720 administration. Transendothelial migration of FTY720-P-treated immature DCs to the CCR7 ligand CCL19 was reduced. These novel data suggest that modulation of DC trafficking by FTY720 may contribute to its immunosuppressive effects. PMID- 16212625 TI - Calcineurin inhibitor effects on growth and phenotype of human airway epithelial cells in vitro. AB - Calcineurin inhibitors (CIs) cyclosporin and tacrolimus form the basis for immunosuppression in lung transplantation, yet also exert biological effects on nonlymphoid tissue. With the advent of inhaled cyclosporin, we hypothesize that the airway epithelium is also subject to CI effects at high doses. The aim of this study was to identify human tracheobronchial epithelial cell (hTBEC) calcineurin gene expression and quantify effects of CIs on hTBEC growth, interleukin-1-beta stimulated IL-8 production and hTBEC phenotype. Cyclophillin B and FK-associated binding protein, calcineurin A (alpha and beta), and NFATC3 and NFAT5 were detected in hTBEC cultures by RT-PCR. Acute and chronic cyclosporine treatment 1000 ng/mL significantly inhibited hTBEC proliferation, while tacrolimus did not (range of 10 ng/mL to 1000 ng/mL for acute treatment, 50 ng/mL for chronic treatment). Cyclosporin at 10,000 ng/mL significantly increased LDH release by well-differentiated hTBEC cultures (n = 6) and trended towards significance at 1000 ng/mL. IL1-beta stimulated IL-8 production was significantly increased in rapidly growing hTBEC cultures (n = 8) treated with cyclosporin (p = 0.049). Prolonged treatment of well-differentiated hTBECs at air-liquid-interface (ALI) with cyclosporin 1000 ng/mL significantly reduced intact multilayered mucociliary epithelium (p = 0.009). Inhibition of hTBEC growth, stimulation of IL 8 production and long-term effects on mucociliary phenotype and intact multi layered epithelium suggest that cyclosporin may have a direct toxic effect on airway epithelium after transplantation. PMID- 16212626 TI - Harmful delayed effects of exogenous isolation enzymes on isolated human islets: relevance to clinical transplantation. AB - The isolation process exposes human pancreatic islets to exogenous isolation enzymes. Exposure to these enzymes, as a result of intraductal injection in the pancreas or simple contact of islets with enzyme components, causes internalization into the islet cells of enzymes and their by-products. Human islets exposed to Liberase-HI exhibit a decreased insulin secretory ability that correlates with the time of exposure. This phenomenon is paralleled by increased expression of adhesion molecules (CD106 and CD62p) and activation of apoptotic pathways (Bax and Bcl-2) in islet cells. Increased functional impairment is also observed after islet transplantation in diabetic immunodeficient mice. Experimental exposure of islet grafts to exogenous isolation enzymes causes intense inflammation (CD11b positive cells) at the transplant site and it was associated with sickness behavior and eventually death of mouse recipients. The extent of these adverse effects likely deceives the standard qualitative protocols currently in use to assess islet quality in vitro. Reducing the secondary effects of exogenous isolation enzymes on isolated human islets may be crucial to enhance the quality of islets as tissue grafts. PMID- 16212627 TI - Kidney transplant in black recipients: are African Europeans different from African Americans? AB - Despite recent improvement, significant racial disparities in outcome still persist after renal transplantation among African American patients in the United States. This study evaluated the association of race and ethnicity with allograft outcomes in a French population of 952 Caucasian (Cauc) patients and 140 African European (AE) patients who underwent renal transplantation in our center between 1987 and 2003. Demographic characteristics were similar for the two cohorts other than cause of end-stage renal failure (more hypertension among AE and more polycystic kidney disease among Cauc) and cold ischemia time (significantly longer for AE). Immunosuppressive treatment was comparable between groups. There were no significant differences between AE and Cauc in the incidence of acute rejection (31% vs. 30%). At 5 years post-transplant, patient survival (93% vs. 92%), graft survival (83% in both groups) and graft function (creatinine clearance 48 mL/min vs. 45 mL/min) were also similar among the AE and Cauc patients. We demonstrate that ethnic origin does not affect outcome after renal transplantation in France. Therefore, differences observed in the United States cannot be only related to immunologic or pharmacologic factors. The results of renal transplantation in patients of African origin could be improved with universal immunosuppressive drug coverage. PMID- 16212628 TI - An evidence-based analysis of simultaneous pancreas-kidney and pancreas transplantation alone. AB - While pancreas transplantation has evolved within two decades from a frustrating and poorly-accepted therapeutic option to a highly successful procedure, the respective benefits of the successive surgical and immunosuppressive developments have remained unclear. The aim of this study was to determine using an evidence based methodology, which novel approaches have contributed to the current results and whether pancreas transplantation is cost-effective. Out of 2481 articles, 102 analyzed either surgical or immunosuppressive aspects of pancreas transplantation. Urological complications were more frequent in bladder over enteric drainage (range: 62-63% vs. 12-20%, p = 0.0001), but without significant difference in patient or graft survival. Portal drainage was associated with a trend toward fewer complications and better hyperinsulinemia control over systemic drainage in retrospective studies. Immunosuppression combining induction therapy, a calcineurin inhibitor, mycophenolate mophetil (MMF) and corticosteroids were associated with a 40% decreased incidence of rejection (p = 0.01) and an increase in graft survival above 90% at 1 year (p < 0.05). Pancreas transplantation is highly cost-effective compared to conservative alternatives. We conclude that despite a paucity of large studies, enteric drainage should be recommended but the benefits of portal venous drainage remain debated. Quadruple immunosuppression protocols including induction therapy should be the standard regimen. PMID- 16212629 TI - Predicting glomerular filtration rate in kidney transplantation: are the K/DOQI guidelines applicable? AB - The kidney disease outcomes quality initiative (K/DOQI) guidelines introduced a classification of chronic kidney disease (CKD) based on the level of kidney function. In order to predict the glomerular filtration rate (GFR), they specifically recommended the use of the modification of diet in renal disease (MDRD) study and Cockcroft-Gault (C-G) equations. Since the performance of these estimates has been questioned, we sought to determine whether these recommendations might be applicable in renal transplantation. Following the K/DOQI methodology, we compared the GFR estimated by the MDRD and C-G equations with 476 inulin clearances performed in 284 renal transplant recipients. Even though the MDRD equations provided a better prediction than C-G formula, none of them reached the level of accuracy required by the K/DOQI standards. At least, 25% of the calculated GFR gave a prediction beyond 30% of the corresponding inulin clearance value. In addition, when classified according to their predicted GFR, less than two-thirds of the transplant patients turned out to be assigned to the correct stage of CKD. We conclude that, in renal transplantation, the predictive performance of both C-G and MDRD study equations appears to be particularly impaired and may potentially compromise the validity of the K/DOQI guidelines if implemented in their current form. PMID- 16212630 TI - Outcome of nonheart-beating donor kidneys with prolonged delayed graft function after transplantation. AB - Nonheart-beating donor (NHBD) kidneys are frequently associated with delayed graft function (DGF), with a deleterious effect on kidney function and allograft survival. The influence and the duration of DGF on the outcome of NHBD kidneys are assessed. All recipients of an NHBD kidney in the period 1993-2003 were reviewed. Excluded from analysis were patients with primary nonfunction (PNF). One hundred and five patients with a functioning NHBD graft were reviewed: 23 (22%) had immediate function (group 1), 40 (38%) had DGF < or = 2 weeks (group 2), 31 (30%) had DGF 15 days to 4 weeks (group 3) and 11 (10%) had DGF for > 4 weeks (group 4). Creatinine clearance at 3 months was higher in groups 1 and 2 versus group 4 (p = 0.015 and p = 0.006, respectively) and was higher in group 2 versus group 4, at 1 year (p = 0.01). Graft survival was 95%, 98%, 97% and 89%, respectively, at 1 year and 95%, 85%, 77% and 89%, respectively, at 5 years, which was not significantly different. The duration of DGF in NHB kidneys has a negative effect on creatinine clearance, but no effect on graft survival. PMID- 16212631 TI - Obesity is associated with worsening cardiovascular risk factor profiles and proteinuria progression in renal transplant recipients. AB - Obesity is associated with adverse cardiovascular (CV) parameters and may be involved in the pathogenesis of allograft dysfunction in renal transplant recipients (RTR). We sought the spectrum of body mass index (BMI) and the relationships between BMI, CV parameters and allograft function in prevalent RTR. Data were collected at baseline and 2 years on 90 RTR (mean age 51 years, 53% male, median transplant duration 7 years), categorized by BMI (normal, BMI < or = 24.9 kg/m2; pre-obese, BMI 25-29.9 kg/m2; obese, BMI > or = 30 kg/m2). Proteinuria and glomerular filtration rate (eGFR(MDRD)) were determined. Nine percent RTR were obese pre-transplantation compared to 30% at baseline (p < 0.001) and follow-up (25 +/- 2 months). As BMI increased, prevalence of metabolic syndrome and central obesity increased (12 vs 48 vs 85%, p < 0.001 and 3 vs 42 vs 96%, p < 0.001, respectively). Systolic blood pressure, fasting blood glucose and lipid parameters changed significantly with BMI category and over time. Proteinuria progression occurred in 65% obese RTR (23 (13-59 g/mol creatinine) to 59 (25-120 g/mol creatinine)). BMI was independently associated with proteinuria progression (beta 0.01, p = 0.008) but not with changing eGFR(MDRD.) In conclusion, obesity is common in RTR and is associated with worsening CV parameters and proteinuria progression. PMID- 16212632 TI - BK virus-specific antibodies and BKV DNA in renal transplant recipients with BKV nephritis. AB - We evaluated twenty renal transplant subjects at various stages of BKV nephritis (BKVN) for BKV-specific IgG and IgM antibodies using ELISA technique and BKV-DNA using PCR. They were divided as early onset (n = 7), stabilizing (n = 3), resolved (n = 8) and late onset (n = 2) BKVN. BKV-specific antibodies and BKV-DNA were simultaneously determined. The mean BKV-specific IgG level in early onset and stabilizing BKVN were 64 and 39 EIA units, and were significantly lower than 138 EIA units seen in resolved BKVN, P = 0.007, P = 0.008. The mean BKV-specific IgM levels in stabilizing BKVN was higher than resolved BKVN (130 vs 51 EIA units), P = 0.006. Mean plasma BKV loads for each group were 955,925, 5642 and 42 copies/mL of plasma, respectively. Prospective study in six BKVN cases revealed mean IgG, IgM levels and BKV-DNA at the time of diagnosis of BKVN as 39, 110 EIA units and 586,758 copies/mL of plasma, respectively. After a mean period of 5.2 months, IgG level increased to 120 EIA units (p = 0.0058) and had no detectable viral copies in circulation. Recovery from BKVN and elimination of BKV is associated with the development of BKV-specific IgG antibodies and this provides insight into the role of humoral immunity to BKV in the pathogenesis of BKVN. PMID- 16212633 TI - Improved long-term outcomes after renal transplantation associated with blood pressure control. AB - Hypertension has a negative impact on long-term outcomes after renal transplantation. We investigated the effect of a recent decline in blood pressure among renal transplant patients in the Collaborative Transplant Study (CTS) database on long-term graft and patient survival. CTS data were used to evaluate transplant outcomes in relation to recipient systolic blood pressure (SBP) for 24,404 first cadaver kidney recipients transplanted between 1987 and 2000. Patients whose SBP was > 140 mmHg at 1 year posttransplantation but controlled to < or = 140 mmHg by 3 years had significantly improved long-term graft outcome compared with patients with sustained high SBP to 3 years (RR 0.79; CI 0.73-0.86; p < 0.001). Additional examination at 5 years showed that SBP lowering after year 3 was associated with improved 10-year graft survival (RR 0.83; CI 0.72-0.96; p = 0.01), whereas even a temporary increase in SBP at 3 years was associated with worse survival (RR 1.37; CI 1.19-1.58; p < 0.001). Changes in SBP were paralleled by changes in the incidence of cardiovascular death among recipients younger than 50 but not in older recipients. Lowering SBP, even after several years of posttransplantation hypertension, is associated with improved graft and patient survival in renal allograft recipients. PMID- 16212634 TI - Effect of pregnancy on long-term kidney function in renal transplant recipients treated with cyclosporine and with azathioprine. AB - In order to investigate the effect of different immunosuppressive regimens and the time interval between transplantation and pregnancy on long-term outcome, we performed a case-control study in pregnant renal allograft recipients. Eighty-one pregnancies of kidney transplanted recipients were identified [cyclosporine (CYA): n = 40; azathioprine (AZA): n = 41]. Controls were matched with respect to important prognostic factors. Posttransplant follow-up was 91.3 +/- 5 months. Graft and patient survival were similar in both groups and there was no apparent effect of immunosuppression. A total of 28 recipients (33%) delivered within 2 years and 6 (8%) subjects within 1 year after transplantation, but these short transplantation-to-pregnancy intervals had no apparent adverse effect on long term outcome. In contrast to AZA-treated patients, CYA-treated patients experienced an increase in serum creatinine postpartum (1.15 +/- 0.2 mg/dL vs. 1.61 +/- 0.1 mg/dL; p < 0.05). Whole blood CYA levels decreased transiently during pregnancy from 115.9 +/- 8 ng/mL to 80.7 +/- 7 ng/mL leading to a gradual increase in drug dose from 240 +/- 14 mg/day to 324 +/- 21 mg/day (p < 0.05). Following delivery, there was an increase in CYA concentrations to 173 +/- 5.4 ng/mL, requiring rapid dose tapering to baseline of 246 +/- 15 mg/day. Pregnancies in renal recipients do not affect long-term patient and graft survival, independent of the immunosuppression. No detrimental effect of short transplantation-to-pregnancy intervals on long-term graft function was detected. PMID- 16212635 TI - Multivariate analysis of risk factors for acute rejection in early corticosteroid cessation regimens under modern immunosuppression. AB - The purpose of this study was to define risk factors for acute rejection with early corticosteroid withdrawal (CSWD; within 7 days posttransplant) in renal transplantation. Data from prospective, IRB-approved early CSWD trials were analyzed. Overall acute rejection rate in 308 patients was 17.1%. Acute rejection rates and observed risks (OR) in patients with individual risk factors were: repeat transplants 38.6%; current PRA >25%; 29.4%; African Americans 23.5%; delayed graft function (DGF) 26.1%; HLA DR mismatches >0 17.9%; female gender 19.7%; Thymoglobulin induction 15.3%; type 1 diabetes 30.8%; type 2 diabetes 11.1%; deceased donor recipients 21%; and living donor recipients 14%. Logistic regression analysis provided the following risks (OR) for acute rejection: repeat transplant 2.51; current PRA > 25% 1.53; African Americans 1.47; DGF 1.58; HLA DR mismatches > 0 1.61; female gender 1.43; Thymoglobulin induction 0.61; type 1 diabetes 2.23, type 2 diabetes 0.5, deceased donor recipients 1.11, and living donor recipients 0.9. Risk factors for acute rejection under early corticosteroid withdrawal are similar to those previously defined under chronic corticosteroid therapy. These observations provide implications for future CSWD trials including: use of T cell depleting antibody induction therapy (thymoglobulin) to reduce acute rejection risk, 2) enrollment stratification for high risk groups, and 3) modified immunosuppression for high risk groups. PMID- 16212636 TI - Expanded criteria donor kidney allocation: marked decrease in cold ischemia and delayed graft function at a single center. AB - Expanded criteria donor (ECD) kidney allocation aims to increase utilization and facilitate placement. We implemented an ECD program for pre-consented candidates and studied whether ECD allocation decreased cold ischemia time and delayed graft function (DGF). We compared donor, recipient and transplant data for ECD transplants performed during the first year of our program to those performed in the preceding 5 1/2 years. Logistic regression identified risk factors for DGF. Of 356 candidates, 107 (30%) consented, 32 (9%) completed evaluation and 20 (6%) underwent ECD transplantation during the program's first year. The recent and historical ECD cohorts had similar donor and recipient characteristics, except that recent ECD recipients were older. The rate of donor kidney biopsy dropped from 85% to 24% (p < 0.001). Cold ischemia time decreased from 16.4 to 7.4 h (p < 0.001), as did the incidence of DGF from 43% to 15% (p = 0.031). Three independent risk factors for DGF emerged: recipient height (OR 1.21/10 cm; p = 0.008), >4 HLA mismatches (OR 20.46; p = 0.0033) and cold ischemia time (OR 1.24/h; p = 0.0036). We conclude the ECD designation provides a description of kidney quality that may obviate biopsy. ECD allocation decreased cold ischemia time and DGF, which may improve graft survival. PMID- 16212637 TI - Living donor liver transplantation for pediatric patients with inheritable metabolic disorders. AB - Forty-six pediatric patients who underwent living donor liver transplantation (LDLT) using parental liver grafts for inheritable metabolic disorders (IMD) were evaluated to determine the outcomes of the surgery, decisive factors for post transplant patient survival and the impact of using donors who were heterozygous for the particular disorder. Disorders included Wilson disease (WD, n = 21), ornithine transcarbamylase deficiency (OTCD, n = 6), tyrosinemia type I (TTI, n = 6), glycogen storage disease (GSD, n = 4), propionic acidemia (PPA, n = 3), methylmalonic acidemia (MMA, n = 2), Crigler-Najjar syndrome type I (CNSI, n = 2), bile acid synthetic defect (BASD, n = 1) and erythropoietic protoporphyria (EPP, n = 1). The post-transplant cumulative patient survival rates were 86.8 and 81.2% at 1 and 5 years, respectively. Post-transplant patient survival and recovery of the growth retardation were significantly better in the liver oriented diseases (WD, OTCD, TTI, CNSI and BASD) than in the non-liver-oriented diseases (GSD, PPA, MMA and EPP) and pre-transplant growth retardation disadvantageously affected post-transplant outcomes. Although 40 of 46 donors were considered heterozygous for each disorder, neither mortality nor morbidity related to the heterozygosis has been observed. LDLT using parental donors can be recommended as an effective treatment for pediatric patients with IMD. In the non liver-oriented diseases, however, satisfactory outcomes were not obtained by hepatic replacement alone. PMID- 16212638 TI - Analysis and outcomes of right lobe hepatectomy in 101 consecutive living donors. AB - The shortage of deceased organ donors has created a need for right lobe living donor liver transplantation (RLDLT) in adults. Concerns regarding donor safety, however, necessitate continuous assessment of donor acceptance criteria and documentation of donor morbidity. We report the outcomes of our first 101 donors who underwent right lobectomy between April 2000 and November 2004. The cohort comprised 58 men and 43 women with a median age of 37.8 years (range: 18.6-55 years); median follow-up is 24 months. The middle hepatic vein (MHV) was taken with the graft in 55 donors. All complications were recorded prospectively and stratified by grade according to Clavien's classification. Overall morbidity rate was 37%; all complications were either grade 1 or 2, and the majority occurred during the first 30 days after surgery. Removal of the MHV did not affect morbidity rate. There were significantly fewer complications in the later half of our experience. All donors are well and have returned to full activities. With careful donor selection and specialized patient care, low morbidity rates can be achieved after right hepatectomy for living donor liver transplantation. PMID- 16212639 TI - Living donor adult liver transplantation: a longitudinal study of the donor's quality of life. AB - We report the results of a prospective, longitudinal quality of life survey on our adult right lobe (RL) liver donors. A total of 47 donors were enrolled; a standard SF-36 form and 43 questions developed by our team were completed before donation, at 1 week, and 1, 3, 6 and 12 months after donation. There were no donor deaths. Twenty-nine complications occurred in 16 patients. Major complication rate was 12.8%. Employment status and personal finances were identified as major stressors. All donors who wished to return to work did so by 1 year (mean 3.4 months). Individuals reported between 0 dollars and 25,000 dollars in losses (wages, travel, lodging, etc.). Relationships with recipients and other family members were not altered significantly. Anticipated pain (predonation) was greater than actual pain reported. Donors indicated satisfaction with the donation process regardless of recipient outcome. Physical complaints were significant at 1 week and 1 month, but returned to baseline. Donor mental health remained stable. In conclusion, RL donors found the experience to be a positive one throughout the first postdonation year. The study identified areas (finances, employment and expected recipient outcomes) to be stressed as future donors are evaluated. PMID- 16212641 TI - IVIG and HLA antibodies. Evidence for inhibition of complement activation but not for anti-idiotypic activity. AB - The immediate effects of IVIG can be due to the presence of anti-idiotypic antibodies or inhibition of complement, but there is limited data about these possible mechanisms specifically on HLA antibodies (HLA Abs). Potential blocking activity of IVIG on HLA Ab binding and complement activation was investigated by flow cytometry. IVIG did not inhibit the IgG binding of any of 23 sera from sensitized patients containing Abs to several different HLA specificities. In contrast, IVIG produced significant dose-dependent complement inhibition. Low IVIG concentrations could be inhibitory if there was little C3 activation, but high concentrations were needed when C3 was activated more efficiently. The data do not support any significant contribution of anti-idiotypic antibodies against HLA Abs to the activity of IVIG. The results also highlight a relationship between the magnitude of C activation and the C inhibitory effect of IVIG. PMID- 16212640 TI - Antibody-mediated rejection in human cardiac allografts: evaluation of immunoglobulins and complement activation products C4d and C3d as markers. AB - Antibody-mediated rejection (AMR) in human heart transplantation is an immunopathologic process in which injury to the graft is in part the result of activation of complement and it is poorly responsive to conventional therapy. We evaluated by immunofluorescence (IF), 665 consecutive endomyocardial biopsies from 165 patients for deposits of immunoglobulins and complement. Diffuse IF deposits in a linear capillary pattern greater than 2+ were considered significant. Clinical evidence of graft dysfunction was correlated with complement deposits. IF 2+ or higher was positive for IgG, 66%; IgM, 12%; IgA, 0.6%; C1q, 1.8%; C4d, 9% and C3d, 10%. In 3% of patients, concomitant C4d and C3d correlated with graft dysfunction or heart failure. In these 5 patients AMR occurred 56-163 months after transplantation, and they responded well to therapy for AMR but not to treatment with steroids. Systematic evaluation of endomyocardial biopsies is not improved by the use of antibodies for immunoglobulins or C1q. Concomitant use of C4d and C3d is very useful to diagnose AMR, when correlated with clinical parameters of graft function. AMR in heart transplant patients can occur many months or years after transplant. PMID- 16212642 TI - Complications related to dapsone use for Pneumocystis jirovecii pneumonia prophylaxis in solid organ transplant recipients. AB - Dapsone, used for prevention of Pneumocystis jirovecii infections, has been reported to cause hemolytic anemia and methemoglobinemia; its tolerability in solid organ transplant recipients is not well described. We investigated dapsone related adverse events in patients undergoing solid organ transplantation from 1999 to 2004. Transplant providers identified patients for the investigators who then reviewed the patients' hospital and outpatient records. Sixteen solid organ transplant recipients fit case definitions for dapsone-related hemolytic anemia (n = 11) or methemoglobinemia (n = 5). Median time from event to dapsone discontinuation was 15 days; all patients improved after drug discontinuation. G6PD enzyme activity was normal in all patients whose test results were available. Dapsone may be associated with hemolytic anemia or methemoglobinemia, even with normal G6PD levels. These events are often not promptly recognized, and drug discontinuation is delayed. Dapsone-related hemolytic anemia or methemoglobinemia should be considered in solid organ transplant recipients with unexplained anemia or hypoxia. PMID- 16212643 TI - Is liver transplantation advisable for isoniazid fulminant hepatitis in active extrapulmonary tuberculosis? AB - Antituberculous treatment is a well-known cause of fulminant hepatic failure (FHF). This could lead to liver transplantation as the only possible treatment, which on the other hand could be contraindicated due to active tuberculosis. The risk of aggressive dissemination of the disease after transplantation is not clearly determined by the current second-line antituberculous therapies. We report a case of vertebral tuberculosis treated with rifampin, isoniazid and pyrazinamide. He developed an FHF that was treated with urgent liver transplantation. Despite the immunosuppression, the disease was well controlled with ciprofloxacin, ethambutol and streptomycin and the patient is in good health 23 months after transplantation. In conclusion, active extrapulmonary tuberculosis should perhaps be considered for liver transplantation when FHF develops due to anti-tuberculous drugs. PMID- 16212644 TI - Sirolimus-induced ulceration of the small bowel in islet transplant recipients: report of two cases. AB - Sirolimus (SRL) has been used for most islet recipients over the past 5 years. It provides balanced immunosuppression in combination with low-dose calcineurin inhibitors, while avoiding corticosteroids. This regimen decreases the risk of nephrotoxicity, neurotoxicity and diabetogenicity. SRL has also been used selectively in clinical liver and kidney transplantation. A number of common side effects including anemia, leucopenia, thrombocytopenia, hypercholesterolemia, mouth ulceration, joint pain, extremity edema and impaired wound healing have been associated with the use of SRL. As SRL is used more frequently, evidence has been gathered on its rare but severe side effects. We report 2 patients who underwent islet transplantation and developed symptomatic small bowel ulceration that resolved after complete withdrawal of SRL. Although small bowel ulceration is rare, it can potentially progress to more serious complications if not treated adequately. Our experience highlights an uncommon but potentially serious adverse effect of high-dose SRL in islet recipients. PMID- 16212647 TI - Uterine response to multiple inoculations with Arcanobacterium pyogenes and Escherichia coli in nulliparous ewes. AB - PROBLEM: Uterine infections seem more severe in nulliparous animals. Our objective was to determine whether intrauterine inoculation of nulliparous ewes with Arcanobacterium pyogenes and Escherichia coli would produce an antibody response and reduce the severity of subsequent infections. METHOD OF STUDY: Nulliparous ewes (n = 9/treatment) received (i) 'primary intrauterine inoculation' with phosphate-buffered saline (PBS) and 'secondary intrauterine inoculation' with PBS; (ii) primary PBS-secondary 75 x 10(7) cfu of A. pyogenes and 35 x 10(7) cfu of E. coli (PBS-Bacteria); (iii) primary bacteria-secondary PBS; or (iv) primary bacteria-secondary bacteria (Bacteria-Bacteria). RESULTS: Inoculations evoked an antibody response. Postmortem examinations 6 days after the secondary inoculation indicated that PBS-treated ewes did not develop uterine infections, but all bacteria-treated ewes did. Infections were either less severe or closer to resolution in Bacteria-Bacteria than they were in PBS-Bacteria ewes. CONCLUSIONS: Intrauterine inoculation of nulliparous ewes with A. pyogenes and E. coli evokes an antibody response that may help the uterus reduce the severity of subsequent infections. PMID- 16212646 TI - Perforin and Fas/FasL cytolytic pathways at the maternal-fetal interface. AB - The immunogenetic enigma of maternal acceptance of the fetal semiallograft has been termed an immunological paradox. The first trimester decidua is heavily infiltrated with CD56(bright) CD16- uterine natural killer (uNK) cells which must be prepared to respond to potential pathogen challenges and still be able to control immune responses that allow the development of the fetus. The significant presence of cytolytic mediators, perforin and Fas/Fas ligand (FasL), at the maternal-fetal interface raises a question of their role(s) in the immunological interrelations between maternal tissues and trophoblast cells. As uNK cells in vitro lyse target cell lines (K562, P815 and P815Fas) using these effector molecules, it seems that, although immunocompetent, their cytotoxicity is not directed against trophoblast during normal pregnancy. Therefore, it is generally believed that the hormonal and Th1/Th2 cytokine balance plays an important role in the tolerance and maintenance of pregnancy. This paper gives an overview of the recent findings on the complex immunological events that occur at the maternal-fetal interface. PMID- 16212648 TI - IgG, IgA and IgM antibodies against FSH: serological markers of pathogenic autoimmunity or of normal immunoregulation? AB - PROBLEM: Autoimmune mechanisms are often involved in causing infertility. Among the possible targets of autoantibodies, the follicle-stimulating hormone (FSH) which regulates the follicular maturation in human ovary is a promising candidate. We aimed to study whether anti-FSH-antibodies might be involved in different clinical types of infertility. METHOD OF STUDY: The study group consisted of 178 patients (75 with polycystic ovary syndrome (PCOS), 103 with endometriosis) and 75 pregnant women. Female blood donors formed the control group (n = 85). Indirect enzyme-linked immunosorbent assay tests were performed using purified FSH as antigens and a synthetic peptide corresponding to the 78-93 region (V14D) of the human FSH beta-chain. CONCLUSION: We showed that anti-FSH antibodies were present in controls and their production decreased during pregnancy. Endometriosis and PCOS were associated with higher values of anti-FSH immunoglobulin (Ig)A, anti-V14D-IgA, and endometriosis with anti-V14D-IgG. Our data suggest that anti-FSH-IgA could be a marker of ovarian disorders that cause infertility. PMID- 16212649 TI - The chemokine SDF-1/CXCL12 contributes to T lymphocyte recruitment in human pre ovulatory follicles and coordinates with lymphocytes to increase granulosa cell survival and embryo quality. AB - We investigated the production and the role of the chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) in pre-ovulatory follicles of women undergoing in vitro fertilization. We detected CXCL12 and its receptor CXCR4 by flow cytometry, western blotting and RT-PCR. We tested cell migration in Transwell experiments. We measured apoptosis using delta psi m-sensitive fluorescent probe DiOC6(3) and we screened apoptosis-related gene expression with macro-arrays. Granulosa cells from follicular aspirates produce CXCL12 that contributes to T lymphocytes recruitment. CXCL12 reduces early apoptosis of granulosa cells. This effect is accompanied by a shift of bcl2/bax ratio, and decreased expression of p53 targeted genes (pig7, pig8, p21, gadd45). Removal of lymphocytes disables CXCL12 mediated anti-apoptotic effect on granulosa cells. Anti-apoptotic activity of CXCL12 is positively correlated to high quality of embryos. In conclusion, CXCL12 is locally produced by luteinizing granulosa cells. It specifically contributes to T lymphocytes recruitment and coordinates with local lymphocytes to increase granulosa cell survival and embryo quality. PMID- 16212650 TI - Lack of soluble TNF-receptors in women with recurrent spontaneous abortion and possibility for its correction. AB - PROBLEM: Tumor necrosis factor (TNF) and soluble TNF receptors (sTNF-Rs) system related with Th1 and Th2 and activity of NF-kappaB/IkappaB regulatory system. This study was designed to compare sTNF-R1 and sTNF-R2 production (shedding) and levels of late activated CD8+ T-lymphocytes in non-pregnant (n = 30) and pregnant (n = 20) normal women and non-pregnant (n = 20) and pregnant (n = 30) RSA women. Effects of progesterone (natural structure) injections in RSA women were studied. METHODS OF STUDY: Levels of sTNF-R1, sTNF-R2, TNF in peripheral blood serum were detected by enzyme-linked immunosorbent assay. Lymphocyte subsets were estimated by multicolor flow cytometry. NK cell cytotoxic activity of peripheral blood lymphocytes (PBL) in whole blood against K562 targets was determined using Europium-release cytotoxicity assay. Mitogen-induced proliferative response of PBL to PHA-P, Con A and PWM were determined by standard 3H-thymidine incorporation assay. RESULTS: Levels of soluble TNF-R1 and TNF-R2 in normal pregnancy were elevated when compared with non-pregnant normal women and pregnant RSA women. Levels of late activated CD8+ T-lymphocytes in normal pregnancy were decreased but no changes were detected in RSA women. After progesterone therapy (i.m. injections of 2.5% oil solution) in RSA women elevation of sTNF-R1 and sTNF R2 to normal pregnancy ranges was observed. No changes in levels of late activated CD8+ T-lymphocytes after progesterone treatment were detected. CONCLUSIONS: Elevation of levels of sTNF-R1, sTNF-R2 and decrease of late activated cytotoxic T-lymphocytes are pronounce markers of normal human pregnancy. In RSA women there are no elevation of sTNF-R1 and sTNF-R2 levels during pregnancy. This deficiency may be restored by progesterone treatment. PMID- 16212651 TI - Immune complex deposition in adult male Sprague-Dawley rats chronically immunized with GnRH. AB - PROBLEM: This study was undertaken to evaluate whether the anti-GnRH antibodies and immune complexes (IC) generated by immunization with GnRH-TT cause cellular damage within the animal. METHOD OF STUDY: Chronic immunization of rats with GnRH TT injected i.m. was followed by tissue/organ analysis for immune complex deposition by immunofluorescence microscopy. Two groups were studied: (1) those immunized throughout the experiment until their ultimate demise, and (2) those given a chance to recover from the effects of chronic immunization before final analysis. RESULTS: GnRH-TT was effective in stopping spermatogenesis, which resumed after withdrawal of the immunogen. Most tissues from chronically immunized animals were not significantly different than controls, however the kidneys of treated animals exhibited a higher accumulation of IC. Despite increased IC deposition, pathologic effects were not detected at the cellular level. CONCLUSIONS: GnRH-TT is an effective immunocontraceptive although the accumulation of glomerular IC represents a potential deleterious side effect. PMID- 16212652 TI - Comparative immunohistochemical study of M-CSF and G-CSF in feto-maternal interface in a multiparity mouse model. AB - PROBLEM: Multiparity status has been found to bring beneficial effects both to the maintenance of pregnancy and to the offspring; however, these effects have not been fully explained. We have previously reported that placentae obtained from multiparous females belonging to a syngeneic mouse crossbreeding showed an important increase in the number of placental macrophages, suggesting that they might constitute a protective subpopulation. Taking into account that macrophage colony stimulating factor (M-CSF) and granulocyte-colony stimulating factor (G CSF) have proved to modulate macrophage activity and that both factors and/or their receptors have been found at feto-maternal interface, in this paper we analyzed the presence of M-CSF and G-CSF in placental tissue employing the same multiparity mouse model in order to investigate the influence of parity status on local immunoregulation factors of macrophage activity. METHOD OF STUDY: Three groups of mice (CBA/J x CBA/J) were analyzed: Primiparous Young, 3.0 +/- 0.5 months old (PY); Primiparous Old, 8.5 +/- 0.5 months old (PO) and Multiparous Old, 8.5 +/- 0.5 months old, with three to four previous pregnancies (MO). The presence of M-CSF and G-CSF in placental tissue was analyzed by immunohistochemistry. Cytokeratin (CK) and vimentin (VIM) expression and PAS staining were also studied. RESULTS: The three groups showed a similar immunostaining pattern for M-CSF in the whole placental trophoblast, while the expression of G-CSF was significantly higher only in the spongy zone in the MO group. Furthermore, all the MO placentae showed 5-11 layers of cells adjacent to the decidua, where G-CSF and M-CSF were highly detected. Conversely, they constituted a thin layer in PY and PO placentae. These cells were proved to be CK(+) and VIM(-) thus demonstrating their trophoblast origin. In addition, the layers closer to the decidua were also PAS+ suggesting that they could be interstitial cells, a type of invading trophoblast. CONCLUSIONS: In our mouse model, we observed an increase in the expression of G-CSF in placental spongiotrophoblast cells in multiparous females, which have been previously proposed as progenitors of the interstitial cells. Furthermore, this is the first report that indicates that parity status increases trophoblast invasion inducing a proliferative effect of the invading cells on the maternal tissue. We suggest that M-CSF and G-CSF secreted by these invading cells could favor the recruitment of macrophages to the trophoblast and might modulate their activity inducing a switch to a protective, non-inflammatory population. PMID- 16212653 TI - Non-classical protein secretion in bacteria. AB - BACKGROUND: We present an overview of bacterial non-classical secretion and a prediction method for identification of proteins following signal peptide independent secretion pathways. We have compiled a list of proteins found extracellularly despite the absence of a signal peptide. Some of these proteins also have known roles in the cytoplasm, which means they could be so-called "moon lightning" proteins having more than one function. RESULTS: A thorough literature search was conducted to compile a list of currently known bacterial non classically secreted proteins. Pattern finding methods were applied to the sequences in order to identify putative signal sequences or motifs responsible for their secretion. We have found no signal or motif characteristic to any majority of the proteins in the compiled list of non-classically secreted proteins, and conclude that these proteins, indeed, seem to be secreted in a novel fashion. However, we also show that the apparently non-classically secreted proteins are still distinguished from cellular proteins by properties such as amino acid composition, secondary structure and disordered regions. Specifically, prediction of disorder reveals that bacterial secretory proteins are more structurally disordered than their cytoplasmic counterparts. Finally, artificial neural networks were used to construct protein feature based methods for identification of non-classically secreted proteins in both Gram-positive and Gram-negative bacteria. CONCLUSION: We present a publicly available prediction method capable of discriminating between this group of proteins and other proteins, thus allowing for the identification of novel non-classically secreted proteins. We suggest candidates for non-classically secreted proteins in Escherichia coli and Bacillus subtilis. The prediction method is available online. PMID- 16212655 TI - Needle and syringe sharing practices of injecting drug users participating in an outreach HIV prevention program in Tehran, Iran: a cross-sectional study. AB - HIV infection rates have reached epidemic proportions amongst injecting drug users (IDUs) in Iran. Although a number of community-based interventions have being implemented in the country, there is little information on the risk behaviors of IDU participants in these programs. This cross-sectional report aimed to compare the risk behaviors of injecting drug users with differential exposure rates to an HIV outreach program in Tehran, Iran. Results indicated that shared use of needle/syringe in the past month was significantly lower among IDUs who received estimated > or = 7 syringes per week than those who did not [adjusted odds ratio (OR) = 14.36, 95% confidence interval (CI) 2.30-89.56]. While the effectiveness of this outreach program needs further evaluation through a longitudinal investigation, our preliminary findings suggest that the outreach program in Tehran may have been beneficial in reducing direct sharing among those who received more than several needles/syringes from the program. PMID- 16212654 TI - Avidin related protein 2 shows unique structural and functional features among the avidin protein family. AB - BACKGROUND: The chicken avidin gene family consists of avidin and several avidin related genes (AVRs). Of these gene products, avidin is the best characterized and is known for its extremely high affinity for D-biotin, a property that is utilized in numerous modern life science applications. Recently, the AVR genes have been expressed as recombinant proteins, which have shown different biotin binding properties as compared to avidin. RESULTS: In the present study, we have employed multiple biochemical methods to better understand the structure-function relationship of AVR proteins focusing on AVR2. Firstly, we have solved the high resolution crystal structure of AVR2 in complex with a bound ligand, D-biotin. The AVR2 structure reveals an overall fold similar to the previously determined structures of avidin and AVR4. Major differences are seen, especially at the 1-3 subunit interface, which is stabilized mainly by polar interactions in the case of AVR2 but by hydrophobic interactions in the case of AVR4 and avidin, and in the vicinity of the biotin binding pocket. Secondly, mutagenesis, competitive dissociation analysis and differential scanning calorimetry were used to compare and study the biotin-binding properties as well as the thermal stability of AVRs and avidin. These analyses pinpointed the importance of residue 109 for biotin binding and stability of AVRs. The I109K mutation increased the biotin-binding affinity of AVR2, whereas the K109I mutation decreased the biotin-binding affinity of AVR4. Furthermore, the thermal stability of AVR2(I109K) increased in comparison to the wild-type protein and the K109I mutation led to a decrease in the thermal stability of AVR4. CONCLUSION: Altogether, this study broadens our understanding of the structural features determining the ligand-binding affinities and stability as well as the molecular evolution within the protein family. This novel information can be applied to further develop and improve the tools already widely used in avidin-biotin technology. PMID- 16212656 TI - Clinical use of coping in affective disorder, a critical review of the literature. AB - BACKGROUND: The relationship between life stressors, coping and affective disorder is interesting when predicting onset of a affective disorder and relapse of mood episodes. METHODS: A literature review of cross-sectional and longitudinal studies concerning coping and affective disorder in adults including a Medline and Embase search was conducted. RESULTS: 11 cross-sectional studies and 17 longitudinal studies concerning affective disorder and coping were found, among these, two studies include patients with bipolar disorder exclusively. Only four studies elucidate whether emotion-oriented and/or avoidance coping styles are associated with a higher risk of developing affective disorder, so this hypothesis remains unclear. Most studies shows that emotion-oriented and avoidance coping strategies are associated with relapse of depressive episodes. Conversely, problem-focused and task-oriented coping seem to be associated with a good outcome. CONCLUSION: There is a gap between coping theory and clinical use of coping and the clinical relevance of coping is, though promising, still unclear. In future research it is recommended to concentrate on development of a semi-structured interview combining coping style, life events and personality traits. PMID- 16212657 TI - Work ethics and general work attitudes in adolescents are related to quality of life, sense of coherence and subjective health - a Swedish questionnaire study. AB - BACKGROUND: Working life is an important arena in most people's lives, and the working line concept is important for the development of welfare in a society. For young people, the period before permanent establishment in working life has become longer during the last two decades. Knowledge about attitudes towards work can help us to understand young people's transition to the labour market. Adolescents are the future workforce, so it seems especially important to notice their attitudes towards work, including attitudes towards the welfare system. The aim of this study was to describe and analyse upper secondary school students' work attitudes, and to explore factors related to these attitudes. METHODS: The sample consisted of 606 upper secondary school students. They all received a questionnaire including questions about quality of life (QOL), sense of coherence (SOC), subjective health and attitudes towards work. The response rate was 91%. A factor analysis established two dimensions of work attitudes. Multivariate analyses were carried out by means of logistic regression models. RESULTS: Work ethics (WE) and general work attitudes (GWA) were found to be two separate dimensions of attitudes towards work. Concerning WE the picture was similar regardless of gender or study programme. Males in theoretical programmes appeared to have more unfavourable GWA than others. Multivariate analyses revealed that good QOL, high SOC and good health were significantly related to positive WE, and high SOC was positively related to GWA. Being female was positively connected to WE and GWA, while studying on a practical programme was positively related to GWA only. Among those who received good parental support, GWA seemed more favourable. CONCLUSION: Assuming that attitudes towards work are important to the working line concept, this study points out positive factors of importance for the future welfare of the society. Individual factors such as female gender, good QOL, high SOC and good health as well as support from both parents, positive experience of school and work contacts related positively to attitudes towards work. Further planning and supportive work have to take these factors into account. PMID- 16212658 TI - The key role for local base order in the generation of multiple forms of China HIV-1 B'/C intersubtype recombinants. AB - BACKGROUND: HIV-1 is a retrovirus with high rate of recombination. Increasing experimental studies in vitro indicated that local hairpin structure of RNA was associated with recombination by favoring RT pausing and promoting strand transfer. A method to estimate the potential to form stem-loop structure by calculating the folding of randomized sequence difference (FORS-D) has been used to investigate the relationship between secondary structure and evolutionary pressure in some genome. It showed that gene regions under strong positive "Darwinian" selection were associated with positive FORS-D values. In the present study, the sequences of HIV-1 subtypes B' and C, both of which represent the parent strains of CRF07_BC, CRF08_BC and China URFs, were selected to investigate the relationship between natural recombination and secondary structure by calculating the FORS-D values. RESULTS: The apparent higher negative FORS-D value region appeared in the gag-pol gene region (nucleotide 0-3000) of HIV-1 subtypes B' and C. Thirteen (86.7%) of 15 mosaic fragments and 17 (81%) of 21 recombination breakpoints occurred in this higher negative FORS-D region. This strongly suggested that natural recombination did not occur randomly throughout the HIV genome, and that there might be preferred (or hot) regions or sites for recombination. The FORS-D analysis of breakpoints showed that most breakpoints of recombinants were located in regions with higher negative FORS-D values (P = 0.0053), and appeared to have a higher negative average FORS-D value than the whole genome (P = 0.0007). The regression analysis also indicated that FORS-D values correlated negatively with breakpoint overlap. CONCLUSION: High negative FORS-D values represent high, base order determined stem-loop potentials and influence mainly the formation of stem-loop structures. Therefore, the present results suggested for the first time that occurrence of natural recombination was associated with high base order-determined stem-loop potential, and that local base order might play a key role in the initiation of natural recombination by favoring the formation of stable stem-loop structures. PMID- 16212659 TI - The role of pro- and anti-inflammatory responses in silica-induced lung fibrosis. AB - BACKGROUND: It has been generally well accepted that chronic inflammation is a necessary component of lung fibrosis but this concept has recently been challenged. METHODS: Using biochemical, histological, immunohistochemistry, and cellular analyses, we compared the lung responses (inflammation and fibrosis) to fibrogenic silica particles (2.5 and 25 mg/g lung) in Sprague-Dawley rats and NMRI mice. RESULTS: Rats treated with silica particles developed chronic and progressive inflammation accompanied by an overproduction of TNF-alpha as well as an intense lung fibrosis. Dexamethasone or pioglitazone limited the amplitude of the lung fibrotic reaction to silica in rats, supporting the paradigm that inflammation drives lung fibrosis. In striking contrast, in mice, silica induced only a limited and transient inflammation without TNF-alpha overproduction. However, mice developed lung fibrosis of a similar intensity than rats. The fibrotic response in mice was accompanied by a high expression of the anti inflammatory and fibrotic cytokine IL-10 by silica-activated lung macrophages. In mice, IL-10 was induced only by fibrotic particles and significantly expressed in the lung of silica-sensitive but not silica-resistant strains of mice. Anti inflammatory treatments did not control lung fibrosis in mice. CONCLUSION: These results indicate that, beside chronic lung inflammation, a pronounced anti inflammatory reaction may also contribute to the extension of silica-induced lung fibrosis and represents an alternative pathway leading to lung fibrosis. PMID- 16212660 TI - Callose (beta-1,3 glucan) is essential for Arabidopsis pollen wall patterning, but not tube growth. AB - BACKGROUND: Callose (beta-1,3 glucan) separates developing pollen grains, preventing their underlying walls (exine) from fusing. The pollen tubes that transport sperm to female gametes also contain callose, both in their walls as well as in the plugs that segment growing tubes. Mutations in CalS5, one of several Arabidopsis beta-1,3 glucan synthases, were previously shown to disrupt callose formation around developing microspores, causing aberrations in exine patterning, degeneration of developing microspores, and pollen sterility. RESULTS: Here, we describe three additional cals5 alleles that similarly alter exine patterns, but instead produce fertile pollen. Moreover, one of these alleles (cals5-3) resulted in the formation of pollen tubes that lacked callose walls and plugs. In self-pollinated plants, these tubes led to successful fertilization, but they were at a slight disadvantage when competing with wild type. CONCLUSION: Contrary to a previous report, these results demonstrate that a structured exine layer is not required for pollen development, viability or fertility. In addition, despite the presence of callose-enriched walls and callose plugs in pollen tubes, the results presented here indicate that callose is not required for pollen tube functions. PMID- 16212661 TI - Two previously proposed P1/P2-differentiating and nine novel polymorphisms at the A4GALT (Pk) locus do not correlate with the presence of the P1 blood group antigen. AB - BACKGROUND: The molecular genetics of the P blood group system and the absence of P1 antigen in the p phenotype are still enigmatic. One theory proposes that the same gene encodes for both the P1 and Pk glycosyltransferases, but no polymorphisms in the coding region of the Pk gene explain the P1/P2 phenotypes. We investigated the potential regulatory regions up- and downstream of the A4GALT (Pk) gene exons. RESULTS: P1 (n = 18) and P2 (n = 9) samples from donors of mainly Swedish descent were analysed by direct sequencing of PCR-amplified 5'- and 3'-fragments surrounding the Pk coding region. Seventy-eight P1 and P2 samples were investigated with PCR using allele-specific primers (ASP) for two polymorphisms previously proposed as P2-related genetic markers (-551_-550insC, 160A>G). Haplotype analysis of single nucleotide polymorphisms was also performed with PCR-ASP. In approximately 1.5 kbp of the 3'-untranslated region one new insertion and four new substitutions compared to a GenBank sequence (AL049757) were found. In addition to the polymorphisms at positions -550 and -160, one insertion, two deletions and one substitution were found in approximately 1.0 kbp of the 5'-upstream region. All 20 P2 samples investigated with PCR-ASP were homozygous for -550insC. However, so were 18 of the 58 P1 samples investigated. Both the 20 P2 and the 18 P1 samples were also homozygous for -160G. CONCLUSION: The proposed P2-specific polymorphisms, -551_-550insC and -160G, found in P2 samples in a Japanese study were found here in homozygous form in both P1 and P2 donors. Since P2 is the null allele in the P blood group system it is difficult to envision how these mutations would cause the P2 phenotype. None of the novel polymorphisms reported in this study correlated with P1/P2 status and the P1/p mystery remains unsolved. PMID- 16212662 TI - Serous borderline tumor of the fallopian tube presented as hematosalpinx: a case report. AB - BACKGROUND: Compared with their ovarian counterparts, serous borderline tumors of the fallopian tube are uncommon, with limited experience about their clinical behaviour. We present a case of serous borderline tumor of the fallopian tube with unusual presentation and summarise all the published cases to date. CASE PRESENTATION: A case of serous borderline tumor of the fallopian tube in a 34 year old patient is presented, incidentally found during routine gynecologic examination. At laparoscopy the tumor was unusually presented as hematosalpinx and was treated by salpingectomy. Cell-cycle analysis of the tumor tissue revealed a diploid DNA content and a low S-phase fraction. There was no evidence of the disease during the follow-up period of 4.6 years. CONCLUSION: The current case and review of the literature suggest salpingectomy as the optimal treatment for patients with serous borderline tumor of the fallopian tube. PMID- 16212663 TI - Branching projections of ventrolateral reticular neurons to the medial preoptic area and lumbo-sacral spinal cord. AB - Different findings indicate that rostral ventrolateral reticular nucleus (RVL) is neuronal substrate of integration and regulation of the cardiovascular functions. Some efferent RVL neurons project to the thoraco-lumbar spinal cord and excite preganglionic sympathetic neurons, to the spinal phrenic motor neurons involved in inspiratory function and increase the activity of vasoconstrictor fibres innervating blood vessels in the skin and skeletal muscle. Our study was aimed at revealing presence of neurons within RVL supplying branching collateral input to the medial preoptic area (MPA) and to the lumbo-sacral spinal cord (SC-L) in the rat. All animal experiments were carried out in accordance with current institutional guidelines for the care and use of experimental animals. We have employed double fluorescent-labelling procedure: the projections were defined by injections of two retrograde tracers: Rhodamine Labelled Bead (RBL) and Fluoro Gold (FG) in the MPA and SC-L, respectively. Our results showed the presence of few single FG neurons and single RBL neurons in the RVL. The size of FG-neurons and RBL-neurons was medium (25-30 microm) and large (50 microm). Few double projecting neurons were distributed in the middle third of RVL nucleus, their size was 30-40 microm. The results demonstrate that pools of neurons in the RVL have collateral projections to the MPA and SC-L and they are involved in ascending and descending pathway. These data suggest that these neurons could play a role in maintaining activity of central and peripheral blood flow. PMID- 16212664 TI - Focal glial activation coincides with increased BACE1 activation and precedes amyloid plaque deposition in APP[V717I] transgenic mice. AB - BACKGROUND: Inflammation is suspected to contribute to the progression and severity of neurodegeneration in Alzheimer's disease (AD). Transgenic mice overexpressing the london mutant of amyloid precursor protein, APP [V717I], robustly recapitulate the amyloid pathology of AD. METHODS: Early and late, temporal and spatial characteristics of inflammation were studied in APP [V717I] mice at 3 and 16 month of age. Glial activation and expression of inflammatory markers were determined by immunohistochemistry and RT-PCR. Amyloid deposition was assessed by immunohistochemistry, thioflavine S staining and western blot experiments. BACE1 activity was detected in brain lysates and in situ using the BACE1 activity kit from R&D Systems, Wiesbaden, Germany. RESULTS: Foci of activated micro- and astroglia were already detected at age 3 months, before any amyloid deposition. Inflammation parameters comprised increased mRNA levels coding for interleukin-1beta, interleukin-6, major histocompatibility complex II and macrophage-colony-stimulating-factor-receptor. Foci of CD11b-positive microglia expressed these cytokines and were neighbored by activated astrocytes. Remarkably, beta-secretase (BACE1) mRNA, neuronal BACE1 protein at sites of focal inflammation and total BACE1 enzyme activity were increased in 3 month old APP transgenic mice, relative to age-matched non-transgenic mice. In aged APP transgenic mice, the mRNA of all inflammatory markers analysed was increased, accompanied by astroglial iNOS expression and NO-dependent peroxynitrite release, and with glial activation near almost all diffuse and senile Abeta deposits. CONCLUSION: The early and focal glial activation, in conjunction with upregulated BACE1 mRNA, protein and activity in the presence of its substrate APP, is proposed to represent the earliest sites of amyloid deposition, likely evolving into amyloid plaques. PMID- 16212665 TI - Directed evolution strategies for improved enzymatic performance. AB - The engineering of enzymes with altered activity, specificity and stability, using directed evolution techniques that mimic evolution on a laboratory timescale, is now well established. However, the general acceptance of these methods as a route to new biocatalysts for organic synthesis requires further improvement of the methods for both ease-of-use and also for obtaining more significant changes in enzyme properties than is currently possible. Recent advances in library design, and methods of random mutagenesis, combined with new screening and selection tools, continue to push forward the potential of directed evolution. For example, protein engineers are now beginning to apply the vast body of knowledge and understanding of protein structure and function, to the design of focussed directed evolution libraries, with striking results compared to the previously favoured random mutagenesis and recombination of entire genes. Significant progress in computational design techniques which mimic the experimental process of library screening is also now enabling searches of much greater regions of sequence-space for those catalytic reactions that are broadly understood and, therefore, possible to model. Biocatalysis for organic synthesis frequently makes use of whole-cells, in addition to isolated enzymes, either for a single reaction or for transformations via entire metabolic pathways. As many new whole-cell biocatalysts are being developed by metabolic engineering, the potential of directed evolution to improve these initial designs is also beginning to be realised. PMID- 16212666 TI - Differing mental health practice among general practitioners, private psychiatrists and public psychiatrists. AB - BACKGROUND: Providing care for mental health problems concerns General Practitioners (GPs), Private Psychiatrists (PrPs) and Public Psychiatrists (PuPs). As patient distribution and patterns of practice among these professionals are not well known, a survey was planned prior to a re-organisation of mental health services in an area close to Paris METHODS: All GPs (n = 492), PrPs (n = 82) and PuPs (n = 78) in the South-Yvelines area in France were informed of the implementation of a local mental health program. Practitioners interested in taking part were invited to include prospectively all patients with mental health problem they saw over an 8-day period and to complete a 6-month retrospective questionnaire on their mental health practice. 180 GPs (36.6%), 45 PrPs (54.9%) and 63 PuPs (84.0%) responded. RESULTS: GPs and PrPs were very similar but very different from PuPs for the proportion of patients with anxious or depressive disorders (70% v. 65% v. 38%, p < .001), psychotic disorders (5% v. 7% v. 30%, p < .001), previous psychiatric hospitalization (22% v. 26 v. 61%, p < .001) and receiving disability allowance (16% v. 18% v. 52%, p < .001). GPs had fewer patients with long-standing psychiatric disorders than PrPs and PuPs (52%, 64% v. 63%, p < .001). Time-lapse between consultations was longest for GPs, intermediate for PuPs and shortest for PrPs (36 days v. 26 v. 18, p < .001). Access to care had been delayed longer for Psychiatrists (PrPs, PuPs) than for GPs (61% v. 53% v. 25%, p < .001). GPs and PuPs frequently felt a need for collaboration for their patients, PrPs rarely (42% v. 61%. v. 10%, p < .001). Satisfaction with mental health practice was low for all categories of physicians (42.6% encountered difficulties hospitalizing patients and 61.4% had patients they would prefer not to cater for). GPs more often reported unsatisfactory relationships with mental health professionals than did PrPs and PuPs (54% v. 15% v. 8%, p < .001). CONCLUSION: GP patients with mental health problems are very similar to patients of private psychiatrists; there is a lack of the collaboration felt to be necessary, because of psychiatrists' workload, and because GPs have specific needs in this respect. The "Yvelines-Sud Mental Health Network" has been created to enhance collaboration. PMID- 16212667 TI - Postoperative IOP prophylaxis practice following uncomplicated cataract surgery: a UK-wide consultant survey. AB - BACKGROUND: In order to minimise postoperative intraocular pressure (IOP) rise, after routine uncomplicated cataract surgery, prophylaxis may be adopted. Currently, there are no specific guidelines in this regard resulting in wide variation in practice across the UK. We sought to document these variations through a questionnaire survey. METHODS: A questionnaire was sent to all consultant ophthalmic surgeons in the UK. RESULTS: 62.6% of surgeons did not use any IOP lowering agents. 37.4% surgeons routinely prescribed some form of medication. The majority (86.8%) used oral diamox. 20.6% of surgeons said they based their practice on evidence, 43.3% on personal experience, and 17.6% on unit policy. Surprisingly, among the two groups of surgeons (those who gave routine prophylaxis, and those who did not) the percentages of surgeons quoting personal experience, unit policy, or presence of evidence was strikingly similar. The timing of the first postoperative IOP check varied from the same day to beyond 2 weeks. Only 20.2% of surgeons had ever seen an adverse event related to IOP rise; this complication is thus very rare. CONCLUSION: This survey highlights a wide variation in the practice and postoperative management of phacoemulsification cataract surgery. What is very striking is that there is a similar proportion of surgeons in the diametrically opposite groups (those who give or do not give routine IOP lowering prophylaxis) who believe that there practice is evidence based. The merits of this study suggests that consideration must be given to drafting a uniform guideline in this area of practice. PMID- 16212669 TI - Evaluation of school absenteeism data for early outbreak detection, New York City. AB - BACKGROUND: School absenteeism data may have utility as an early indicator of disease outbreaks, however their value should be critically examined. This paper describes an evaluation of the utility of school absenteeism data for early outbreak detection in New York City (NYC). METHODS: To assess citywide temporal trends in absenteeism, we downloaded three years (2001-02, 2002-03, 2003-04) of daily school attendance data from the NYC Department of Education (DOE) website. We applied the CuSum method to identify aberrations in the adjusted daily percent absent. A spatial scan statistic was used to assess geographic clustering in absenteeism for the 2001-02 academic year. RESULTS: Moderate increases in absenteeism were observed among children during peak influenza season. Spatial analysis detected 790 significant clusters of absenteeism among elementary school children (p < 0.01), two of which occurred during a previously reported outbreak. CONCLUSION: Monitoring school absenteeism may be moderately useful for detecting large citywide epidemics, however, school-level data were noisy and we were unable to demonstrate any practical value in using cluster analysis to detect localized outbreaks. Based on these results, we will not implement prospective monitoring of school absenteeism data, but are evaluating the utility of more specific school-based data for outbreak detection. PMID- 16212668 TI - Patients' perspectives on how idiopathic pulmonary fibrosis affects the quality of their lives. AB - BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a debilitating lung disease with a survival of only three to five years from the time of diagnosis. Due to a paucity of studies, large gaps remain in our understanding of how IPF affects the quality of patients' lives. In only one other study did investigators ask patients directly for their perspectives on this topic. Further, currently there is no disease-specific instrument to measure health-related quality of life (HRQL) in patients with IPF. A carefully constructed measurement instrument, sensitive to underlying change, is needed for use in clinical trials and longitudinal studies of patients with IPF. Before developing such an instrument, researchers must improve their understanding of the relevant effects of IPF on patients' lives. On a broader scale, to provide the best care for people with IPF, clinicians must appreciate--from patients' perspectives--how this disease affects various aspects of their lives. METHODS: We used focus groups and individual in-depth interviews with 20 IPF patients to collect their perspectives on how IPF affects their lives (with a focus on the quality of their lives). We then analyzed these perspectives and organized them into a conceptual framework for describing HRQL in patients with IPF. Next, we examined how well certain existing measurement instruments--which have been administered to IPF patients in prior studies--covered the domains and topics our patients identified. RESULTS: In our framework, we identified 12 primary domains: symptoms, IPF therapy, sleep, exhaustion, forethought, employment and finances, dependence, family, sexual relations, social participation, mental and spiritual well-being, mortality. Each domain is composed of several topics, which describe how IPF affects patients' lives. When we compared the content of our conceptual framework with the existing instruments, we found the coverage of the existing instruments to be inadequate for several reasons, including they may tap general areas of QOL or HRQL but not some areas that appear to be most directly affected by IPF, and they include items that are relevant to symptoms and effects of other respiratory diseases but not IPF. CONCLUSION: Collecting patients' perspectives and developing an organized inventory of the relevant effects of IPF on patients' lives provides valuable information for improving our understanding of the impact of this disease on patients and their loved ones. We believe our findings will help alert clinicians and researchers to IPF patients' experiences and concerns. Based on the comparison or our conceptual framework with the content of four existing instruments, it would appear that developing an IPF-specific measurement instrument is justified. Our conceptual framework for describing health-related quality of life in patients with IPF lays a solid foundation for constructing such an instrument. PMID- 16212670 TI - Modeling the effect of age in T1-2 breast cancer using the SEER database. AB - BACKGROUND: Modeling the relationship between age and mortality for breast cancer patients may have important prognostic and therapeutic implications. METHODS: Data from 9 registries of the Surveillance, Epidemiology, and End Results Program (SEER) of the United States were used. This study employed proportional hazards to model mortality in women with T1-2 breast cancers. The residuals of the model were used to examine the effect of age on mortality. This procedure was applied to node-negative (N0) and node-positive (N+) patients. All causes mortality and breast cancer specific mortality were evaluated. RESULTS: The relationship between age and mortality is biphasic. For both N0 and N+ patients among the T1-2 group, the analysis suggested two age components. One component is linear and corresponds to a natural increase of mortality with each year of age. The other component is quasi-quadratic and is centered around age 50. This component contributes to an increased risk of mortality as age increases beyond 50. It suggests a hormonally related process: the farther from menopause in either direction, the more prognosis is adversely influenced by the quasi-quadratic component. There is a complex relationship between hormone receptor status and other prognostic factors, like age. CONCLUSION: The present analysis confirms the findings of many epidemiological and clinical trials that the relationship between age and mortality is biphasic. Compared with older patients, young women experience an abnormally high risk of death. Among elderly patients, the risk of death from breast cancer does not decrease with increasing age. These facts are important in the discussion of options for adjuvant treatment with breast cancer patients. PMID- 16212671 TI - Retroperitoneal inflammatory myofibroblastic tumor. AB - BACKGROUND: Inflammatory myofibroblastic tumor (IMT) is a neoplasm of unknown etiology occurring at various sites. By definition, it is composed of spindle cells (myofibroblasts) with variable inflammatory component, hence the name is IMT. CASE PRESENTATION: The present case is of a 46 years old woman presented with a history of flank pain, abdominal mass and intermittent hematuria for last 6 months. The initial diagnosis was kept as renal cell carcinoma. Finally, it turned out to be a case of retroperitoneal IMT. The patient was managed by complete surgical resection of the tumor. CONCLUSION: IMT is a rare neoplasm of uncertain biological potential. Complete surgical resection remains the mainstay of the treatment. PMID- 16212672 TI - Iranian staff nurses' views of their productivity and human resource factors improving and impeding it: a qualitative study. AB - BACKGROUND: Nurses, as the largest human resource element of health care systems, have a major role in providing ongoing, high-quality care to patients. Productivity is a significant indicator of professional development within any professional group, including nurses. The human resource element has been identified as the most important factor affecting productivity. This research aimed to explore nurses' perceptions and experiences of productivity and human resource factors improving or impeding it. METHOD: A qualitative approach was used to obtain rich data; open, semi-structured interviews were also conducted. The sampling was based on the maximum variant approach; data analysis was carried out by content analysis, with the constant comparative method. RESULTS: Participants indicated that human resources issues are the most important factor in promoting or impeding their productivity. They suggested that the factors influencing effectiveness of human resource elements include: systematic evaluation of staff numbers; a sound selection process based on verifiable criteria; provision of an adequate staffing level throughout the year; full involvement of the ward sister in the process of admitting patients; and sound communication within the care team. Paying attention to these factors creates a suitable background for improved productivity and decreases negative impacts of human resource shortages, whereas ignoring or interfering with them would result in lowering of nurses' productivity. CONCLUSION: Participants maintained that satisfactory human resources can improve nurses' productivity and the quality of care they provide; thereby fulfilling the core objective of the health care system. PMID- 16212673 TI - Patterns and predictors of place of cancer death for the oldest old. AB - BACKGROUND: Cancer patients increasingly are among older age groups, but to date little work has examined the trends in cancer among older people, particularly in relation to end of life care and death. This study describes the older population who die of cancer and the factors which may affect their place of death. METHODS: A Cross-sectional analysis of national data was performed. The study included all people aged 75 and over dying of cancer in England and Wales between 1995 and 1999. The population was divided into exclusive 5 year age cohorts, up to 100 years and over. Descriptive analysis explored demographic characteristics, cancer type and place of death. RESULTS: Between 1995 and 1999, 315,462 people aged 75 and over were registered as dying from cancer. The number who died increased each year slightly over the 5 year period (1.2%). In the 75-79 age group, 55 % were men, in those aged 100 and over this fell to 16%. On reaching their hundreds, the most common cause of death for men was malignancies of the genital organs; and for women it was breast cancer. The most frequent place of death for women in their hundreds was the care home; for men it was hospitals. Those dying from lymphatic and haematopoietic malignancies were most likely to die in hospitals, those with head and neck malignancies in hospices and breast cancer patients in a care home. CONCLUSION: The finding of rising proportions of cancer deaths in institutions with increasing age suggests a need to ensure that appropriate high quality care is available to this growing section of the population. PMID- 16212674 TI - Australian health policy on access to medical care for refugees and asylum seekers. AB - Since the tightening of Australian policy for protection visa applicants began in the 1990s, access to health care has been increasingly restricted to asylum seekers on a range of different visa types. This paper summarises those legislative changes and discusses their implications for health policy relating to refugees and asylum seekers in Australia. Of particular concern are asylum seekers on Bridging Visas with no work rights and no access to Medicare. The paper examines several key questions: What is the current state of play, in terms of health screening and medical care policies, for asylum seekers and refugees? Relatedly, how has current policy changed from that of the past? How does Australia compare with other countries in relation to health policy for asylum seekers and refugees? These questions are addressed with the aim of providing a clear description of the current situation concerning Australian health policy on access to medical care for asylum seekers and refugees. Issues concerning lack of access to appropriate health care and related services are raised, ethical and practical issues are explored, and current policy gaps are investigated. PMID- 16212675 TI - Asthma and COPD in cystic fibrosis intron-8 5T carriers. A population-based study. AB - BACKGROUND: Carriers of cystic fibrosis intron-8 5T alleles with high exon-9 skipping could have increased annual lung function decline and increased risk for asthma or chronic obstructive pulmonary disease (COPD). METHODS: We genotyped 9131 individuals from the adult Danish population for cystic fibrosis 5T, 7T, 9T, and F508del alleles, and examined associations between 11 different genotype combinations, and annual FEV1 decline and risk of asthma or COPD. RESULTS: 5T heterozygotes vs. 7T homozygous controls had no increase in annual FEV1 decline, self-reported asthma, spirometry-defined COPD, or incidence of hospitalization from asthma or COPD. In 5T/7T heterozygotes vs. 7T homozygous controls we had 90% power to detect an increase in FEV1 decline of 8 ml, an odds ratio for self reported asthma and spirometry-defined COPD of 1.9 and 1.7, and a hazard ratio for asthma and COPD hospitalization of 1.8 and 1.6, respectively. Both 5T homozygotes identified in the study showed evidence of asthma, while none of four 5T/F508del compound heterozygotes had severe pulmonary disease. 7T/9T individuals had annual decline in FEV1 of 19 ml compared with 21 ml in 7T homozygous controls (t-test: P = 0.03). 6.7% of 7T homozygotes without an F508del allele in the cystic fibrosis transmembrane conductance regulator gene reported asthma vs. 11% of 7T/9T individuals with an F508del allele (chi2: P = 0.01) and 40% of 7T homozygotes with an F508del allele (P = 0.04). 7T homozygotes with vs. without an F508del allele also had higher incidence of asthma hospitalization (log-rank: P = 0.003); unadjusted and adjusted equivalent hazard ratios for asthma hospitalization were 11 (95%CI: 1.5-78) and 6.3 (0.84-47) in 7T homozygotes with vs. without an F508del allele. CONCLUSION: Polythymidine 5T heterozygosity is not associated with pulmonary dysfunction or disease in the adult Caucasian population. Furthermore, our results support that F508del heterozygosity is associated with increased asthma risk independently of the 5T allele. PMID- 16212676 TI - Development of depression: sex and the interaction between environment and a promoter polymorphism of the serotonin transporter gene. AB - Previous research has demonstrated that a polymorphism in the serotonin transporter gene (5-HTTLPR) and adverse psychosocial circumstances interact to predict depression. The purpose of the present study was to explore the extent to which sex modulates these effects. Eighty-one boys and 119 girls (16-19 years old) were interviewed about psychosocial background variables and genotyped for the 5-HTT promoter polymorphism. There were two main results. First, boys and girls carrying the short 5-HTTLPR allele react to different kinds of environmental factors. Whereas males were affected by living in public housing rather than in own owned homes and by living with separated parents, females were affected by traumatic conflicts within the family. Second, the responses of males and females carrying the short 5-HTTLPR allele to environmental stress factors go in opposite directions. Thus, whereas females tend to develop depressive symptoms, males seem to be protected from depression. The results suggest that both the molecular and the psychosocial mechanisms underlying depression may differ between boys and girls. PMID- 16212677 TI - The impact of irrelevant dimensional variation on rule-based category learning in patients with Parkinson's disease. AB - This study examined the impact of irrelevant dimensional variation on rule-based category learning in patients with Parkinson's disease (PD), older controls (OC), and younger controls (YC). Participants were presented with 4-dimensional, binary valued stimuli and were asked to categorize each into 1 of 2 categories. Category membership was based on the value of a single dimension. Four experimental conditions were administered in which there were zero, 1, 2, or 3 randomly varying irrelevant dimensions. Results indicated that patients with PD were impacted to a greater extent than both the OC and YC participants when the number of randomly varying irrelevant dimensions increased. These results suggest that the degree of working memory and selective attention requirements of a categorization task will impact whether PD patients are impaired in rule-based category learning, and help to clarify recent discrepancies in the literature. PMID- 16212678 TI - MS patients with depressive symptoms exhibit affective memory biases when verbal encoding strategies are suppressed. AB - As many as 50% of multiple sclerosis (MS) patients experience clinical or subclinical depression. A voluminous literature has documented affective memory biases (AMB) among depressed individuals. Despite this, little is known regarding how depressive symptoms may affect MS patients' ability to recall positive and negative material. The present study employed an affective list-learning task that increased cognitive load and inhibited the use of higher order encoding strategies. The purpose of the study was twofold: to determine whether MS patients exhibit AMB and to examine whether subvocal repetition and other higher order encoding strategies are essential to the formation of AMB among people experiencing depression. Results indicated a strong relationship between depression and AMB in MS. The results are discussed in relation to existing biological research that indicates limbic and/or other subcortical systems may play a role in the formation of AMB. PMID- 16212679 TI - Neurological, neuropsychological, and functional outcome following treatment for unruptured intracranial aneurysms. AB - The objective of this study was to carry out a detailed investigation of the neurological, neuropsychological, and return-to-work status of treatment for unruptured intracranial aneurysms (UIAs). A prospective design was used to evaluate the outcome of UIA treatment in a group of 26 UIA patients. Over a 24 month period UIA patients were assessed prior to treatment, during hospitalization, at three months and at six months following treatment. Their performance was compared to a group of 20 matched controls. Neurological morbidity as a result of the UIA treatment was 5%, as assessed by the Glasgow Outcome Scale (GOS) or Rankin at 3 months. The Telephone Interview for Cognitive Status (TICS) proved to be unreliable as a measure of cognitive change. Reliability of change analysis was more sensitive than group analysis, and revealed a pattern of cognitive deficits in 10% of patients as a result of the UIA treatment. In addition, 25% of patients reported a change in work role as a result of the UIA treatment. While 10% of patients sustained mild to moderate neurological and cognitive impairments 3 to 6 months following UIA treatment, their deficits were not as wide-ranging nor as severe as those sustained by patients who survive a subarachnoid hemorrhage (SAH). PMID- 16212680 TI - Anxiety and depression, attention, and executive functions in hypothyroidism. AB - BACKGROUND: Divergences in cognitive disturbances in hypothyroidism reported in the literature are a result of a methodological bias. METHODS: By using a precise methodology, we examined attention and executive functions in hypothyroidism, verified the presence of anxiety and depressive symptoms in hypothyroidism, and examined the possible link between these symptoms and the cognitive disturbances (searching for attentional bias for words with a negative emotional valence). We administered a battery of cognitive tests to 23 participants who had undergone thyroidectomy for thyroid carcinoma: for the first time in an euthyroid state, then 3 weeks later (still in the euthyroid state) to assess the test/retest effect, and finally 4 weeks later in an hypothyroid state. We compared their performance with that of a group of 26 control participants who were also administered the same cognitive tests, also 3 times. RESULTS: In hypothyroidism, the thyroid participants were more anxious and depressed than the controls and presented attentional and executive disturbances that reflected general slowing and difficulties in using their capacities of inhibition. However, they did not exhibit an attentional bias for words with a negative emotional valence. CONCLUSIONS: Contrary to what was expected, symptoms of anxiety and not symptoms of depression interfered with the cognitive performance of participants in hypothyroidism. PMID- 16212681 TI - Further evidence for a comparable memory advantage of self-performed tasks in Korsakoff's syndrome and nonamnesic control subjects. AB - Two experiments were carried out to examine memory in persons with amnesia using self-performed tasks. In Experiment 1, persons with Korsakoff's syndrome and nonamnesic participants with alcoholism learned action phrases not involving real objects by either self-performed tasks or verbal tasks. As indexed by free recall and recognition tests, a memory advantage favoring self-performed tasks was confirmed in both participant groups. In Experiment 2, persons with Korsakoff's syndrome, nonamnesic alcoholic participants, and young control participants learned object names under three different study conditions that differed from one another as to whether actions for each name were verbally generated and whether actions actually were performed. Verbal generation with or without performing the action facilitated recognition, whereas recall advantage was found only in the verbal generation-plus performance condition. These findings confirm a comparable memory advantage of self-performed tasks for a group with Korsakoff's syndrome and a group of nonamnesic comparison participants. Action memory therefore has potential therapeutic implications for memory rehabilitation. PMID- 16212682 TI - Visuomotor control in survivors of childhood acute lymphoblastic leukemia treated with chemotherapy only. AB - Treatment for childhood acute lymphoblastic leukemia (ALL), which includes CNS prophylaxis, is associated with central and peripheral neurotoxicity. The purpose of the present study was to analyze the effects of chemotherapy on various levels of visuomotor control in survivors of childhood ALL treated without cranial irradiation, and to identify risk factors for possible deficits. Visuomotor function was compared between children after treatment for ALL (n = 34), children after treatment for Wilms tumor, which consists of non-CNS directed chemotherapy (n = 38), and healthy controls (n = 151). Three tasks were administered: a simple visual reaction time task and two tasks measuring visuomotor control with one requiring a higher level of cognitive control than the other. Visuomotor deficits were detected only in the ALL group, with poorer performance restricted to the condition requiring the highest level of control. Significant risk factors for poorer performance were female gender and a short time since end of treatment, and a trend was found for a young age at diagnosis. A high cumulative methotrexate dose was an adverse predictive factor in girls. The results indicate that chemotherapy-induced central neurotoxicity in childhood ALL treatment is associated with higher order visuomotor control deficits. Girls appear to be particularly vulnerable. PMID- 16212683 TI - Priming for novel between-word associations in patients with organic amnesia. AB - Ten amnesic patients of various etiologies and 10 matched normal controls participated in this study. On 2 consecutive days, subjects studied 30 novel word word associations 6 times. Using a cued recall task, we assessed episodic learning and delayed retention of the study material immediately after each study phase and again 24 hr after the final study phase. Further, we evaluated implicit memory for new between-word associations by means of an automatic relational priming paradigm immediately after the delayed cued recall trial. Amnesic patients performed poorly on the cued recall task. Moreover, in the overall group of amnesics the priming effect failed to reach statistical significance. When the overall group of amnesics was split according to mean performance on the cued recall task, those in the low performer subgroup--comprised of 6 patients with direct or indirect involvement of the hippocampi--were particularly poor at episodically remembering the associations and did not reveal any relational priming. These data support the hypothesis of similar impairment of new episodic and implicit learning in amnesic patients and suggest that the hippocampus is crucial for both kinds of new learning. PMID- 16212684 TI - Interpreting patient/informant discrepancies of reported cognitive symptoms in MS. AB - Although numerous studies have shown that brain-damaged patients tend to underestimate neuropsychological (NP) impairment when self-ratings are compared to informant ratings, the meaning of such discrepancies is not well studied in multiple sclerosis (MS). We compared patient self- and informant-report questionnaire ratings of NP functioning in 122 MS patients and 37 age- and education-matched normal controls. In addition to completing the Multiple Sclerosis Neuropsychological Questionnaire (MSNQ), participants underwent NP testing and assessment of depression, personality, and neuropsychiatric symptoms. Based on the normal distribution of discrepancy scores, patients were classified according to whether they overestimated or underestimated their cognitive ability, relative to informant ratings. ANOVAs comparing test scores derived from overestimators, underestimators, and accurate estimators were significant for multiple measures of cognitive function, depression, personality, and neuropsychiatric symptoms. Overestimators were characterized by less depression and conscientiousness, and greater degrees of cognitive impairment, euphoric behavioral disinhibition, and unemployment as compared to underestimators. We conclude that patient/informant discrepancy scores on the MSNQ are associated with the aforementioned neuropsychiatric features, and that the MSNQ has potential utility for predicting euphoria and disinhibition syndromes in MS. PMID- 16212685 TI - Cognitive ability predicts degree of genetic abnormality in participants with 18q deletions. AB - One of the most common chromosomal deletions is a loss of genetic material from the long arm of chromosome 18. Most individuals with this condition exhibit mental retardation (68%), yet previous attempts to link cognitive status to deletion size have not shown an association, possibly because cases with additional genetic abnormalities were included. We studied 46 participants ranging from 3 to 35 years of age who had a pure genetic abnormality by excluding those with mosaicism or complex genetic rearrangements. Our patients had terminal deletions ranging from a proximal breakpoint at 18q21.1 (greater genetic abnormality, larger deletion size) to a more distal breakpoint at 18q23 characterized with molecular genetic techniques. Cognitive ability, assessed with the age-appropriate measure (Bayley, 1993 , Differential Ability Scale, Wechsler Scales), ranged from IQ = 49 to 113, with a predominance of mild and moderate mental retardation. Using multivariate regression, deletion size breakpoint rank order was predicted by cognitive ability, age, and adaptive behavior (Vineland Adaptive Behavior Scales), accounting for 36% of the variance in deletion size. However, lower cognitive ability (beta = .34, p = .032) and younger age (beta = .296, p = .024) predicted a larger deletion size, but adaptive behavior (beta = .225, p = .15) did not. An additional multivariate regression showed that cognitive ability and age together accounted for 33% of the variance in deletion size, whereas univariate regression showed that cognitive ability accounted for 26% of the variance and age accounted for 11% of the variance. These findings suggest that degree of cognitive impairment is associated with genetic abnormality when a large sample of individuals with "pure" deletions of genetic material from chromosome 18 is examined. PMID- 16212686 TI - Parkinsonian signs and cognitive function in old age. AB - Studies have shown that parkinsonian signs are related to cognitive function in aging. What remains unclear is whether this association is stronger for some cognitive domains than it is for others, and precisely how much variability in global and specific cognitive functions is explained by the motor signs. We examined the associations between four parkinsonian signs (gait, rigidity, bradykinesia, tremor) and five cognitive domains (episodic memory, semantic memory, working memory, perceptual speed, visuospatial ability) in a large cohort of older persons who were free of Parkinson's disease and dementia and were participating in the Rush Memory and Aging Project. In a series of regression equations that controlled for age, sex, and education, higher levels of three signs (gait, rigidity, and bradykinesia) were related to lower levels of cognitive function, but they accounted for less than 5% of the variance in most measures. The results did not change when the presence of depressive symptoms, diabetes, and hypertension were added to the models. The cross-sectional association between parkinsonian signs and cognitive function did not vary substantially across specific cognitive domains or specific cognitive tests. The results suggest that parkinsonian signs have a modest, but statistically reliable, association with level of cognitive function in old age. PMID- 16212687 TI - The Stick Design test: a new measure of visuoconstructional ability. AB - Visuoconstructional ability is an important domain for assessment in dementia. Use of graphomotor measures dominate this area; however, participants with low education produce results that cannot be easily interpreted. Our objective was to develop and validate a nongraphomotor assessment of visuoconstructional ability for use in dementia evaluations in persons with low or no education. In a longitudinal, population-based study of dementia among Yoruba residents of Ibadan, Nigeria aged 65 years and older, participants underwent clinical assessment with a battery of cognitive tests and consensus diagnosis. Performance on two visuoconstructional tests, Constructional Praxis and Stick Design, were compared. Gender, age, and education affected performance on both tests. The Stick Design test was more acceptable than Constructional Praxis as measured by the number of participants with total test failure (3.9% vs. 15.1%). The Stick Design test was significantly more sensitive to cognitive impairment and dementia than the Constructional Praxis test. We conclude that Stick Design is a reasonable test of visuoconstructional ability in older cohorts with very limited educational exposure and literacy. PMID- 16212688 TI - Assessment of executive dysfunction following traumatic brain injury: comparison of the BADS with other clinical neuropsychological measures. AB - Traditional neuropsychological measures of executive dysfunction (ED) are widely believed to lack adequate sensitivity and selectivity. This may indicate that existing measures are poorly designed and constructed, although an alternative explanation is that executive cognition is multifactorial, such that its assessment necessarily requires administration of multiple measures. This possibility led to the development of a test battery, the Behavioural Assessment of the Dysexecutive Syndrome (BADS). To investigate the sensitivity of the BADS to ED, it and various other measures of ED were administered to 64 persons who had sustained traumatic brain injury. The treating clinical neuropsychologist and occupational therapist for each participant also completed a behavioural rating scale, the Dysexecutive Questionnaire (DEX). Four factors were found to underlie scores on the neuropsychological measures, but few tests were sufficiently powerful to make a significant unique contribution to predicting scores on the DEX. This confirms that multiple tests, drawn from both the BADS and other sources, may be necessary to detect ED in a clinical population. PMID- 16212689 TI - Perceived cognitive dysfunction and observed neuropsychological performance: longitudinal relation in persons with multiple sclerosis. AB - The relation between self-reported cognitive dysfunction and neuropsychological performance over 24 weeks was assessed in a sample of 53 multiple sclerosis patients. Subjects were assessed at Weeks Zero and 24 as part of a clinical trial to enhance cognition. At baseline, subjects had at least mild cognitive impairment on the Rey Auditory Verbal Learning Test and an absence of depression. Neuropsychological performance was assessed with a modification of the well standardized Brief Repeatable Battery. The 5-item Perceived Deficits Questionnaire and a 2-item memory and attention/concentration questionnaire assessed self-perceived cognitive impairment. Self-assessed cognition did not correlate with neuropsychological performance at either baseline or 24 weeks. However, changes in the self-assessment measures did correlate with changes in neuropsychological performance. Patients accurately perceived some changes in their level of cognitive dysfunction, though they were insensitive to the degree of their current dysfunction. Possible explanations of this pattern of results are discussed. PMID- 16212690 TI - Criterion-referenced validity of a neuropsychological test battery: equivalent performance in elderly Hispanics and non-Hispanic Whites. AB - This study examined the validity of the Spanish and English Neuropsychological Assessment Scales (SENAS) in comparison with clinical diagnosis of normal cognition versus cognitive impairment, not demented (CIND) versus demented in elderly Hispanics and Whites. Relationships between SENAS scales and diagnosis were essentially the same in Hispanics and Whites. Verbal memory measures were most strongly related, with more than 35% of the variance in these measures accounted for by diagnosis independent of effects of education, age, gender, and language. Diagnosis accounted for more than 10% of the variance (19% on average) in 11 of the 17 measures examined in this study. Logistic regressions showed that verbal memory was important both for distinguishing normal from CIND and CIND from demented. Object naming improved discrimination of CIND from demented beyond that of verbal memory alone. These results provide evidence of equivalent validity across Hispanics and Whites. PMID- 16212692 TI - Differences in attention, executive functioning, and memory in children with and without ADHD after severe traumatic brain injury. AB - Although the development of Attention Deficit Hyperactivity Disorder (ADHD) after traumatic brain injury (TBI) has been described, it is unknown whether children with TBI and ADHD have greater neuropsychological impairments than children with TBI alone. This study examines attention, executive functioning, and memory in children with TBI-only and TBI + ADHD. Caregivers of 82 children with severe TBI completed structured psychiatric interviews at enrollment to diagnose premorbid ADHD and one-year after injury to diagnose post-injury ADHD. Children underwent neuropsychological testing one year after injury. One memory measure significantly differentiated children with TBI-only from children with newly developed ADHD [secondary ADHD (S-ADHD)] and those with premorbid ADHD that persisted after injury [persisting ADHD (P-ADHD)]. Compared with the TBI-only group, children with TBI + ADHD had worse performance on measures of attention, executive functioning, and memory. Results reveal that in children with severe TBI, the behavioral diagnosis of ADHD is associated with more difficulty in attention, executive functioning, and memory. Additionally, results suggest greater deficits in memory skills in the S-ADHD group compared with the P-ADHD group. Although findings provide preliminary support for distinguishing P-ADHD from S-ADHD, further research is needed to investigate neuropsychological differences between these subgroups of children with severe TBI. PMID- 16212691 TI - fMRI reveals alteration of spatial working memory networks across adolescence. AB - Recent studies have described neuromaturation and cognitive development across the lifespan, yet few neuroimaging studies have investigated task-related alterations in brain activity during adolescence. We used functional magnetic resonance imaging (fMRI) to examine brain response to a spatial working memory (SWM) task in 49 typically developing adolescents (25 females and 24 males; ages 12-17). No gender or age differences were found for task performance during SWM. However, age was positively associated with SWM brain response in left prefrontal and bilateral inferior posterior parietal regions. Age was negatively associated with SWM activation in bilateral superior parietal cortex. Gender was significantly associated with SWM response; females demonstrated diminished anterior cingulate activation and males demonstrated greater response in frontopolar cortex than females. Our findings indicate that the frontal and parietal neural networks involved in spatial working memory change over the adolescent age range and are further influenced by gender. These changes may represent evolving mnemonic strategies subserved by ongoing adolescent brain development. PMID- 16212696 TI - Remembering Bob Rodieck: 1937-2003. AB - In this special issue of Visual Neuroscience , we present a series of papers to honor the life and career of Robert William Rodieck, who passed away at his home in Seattle on September 30, 2003. Rodieck held the E.K. Bishop Professorship in Ophthalmology at the University of Washington Medical Center from 1978-1997. Known to everyone as "Bob," he leaves behind an intellectual legacy often admired by his colleagues and friends for its scope, intensity, and empathy for what was beautiful in the object of his studies. PMID- 16212697 TI - Wide-field ganglion cells in macaque retinas. AB - To describe the wide-field ganglion cells, they were injected intracellularly with Neurobiotin using an in vitro preparation of macaque retina and labeled with streptavidin-Cy3. The retinas were then labeled with antibodies to choline acetyltransferase and other markers to indicate the depth of the dendrites within the inner plexiform layer (IPL) and analyzed by confocal microscopy. There were eight different subtypes of narrowly unistratified cells that ramified in each of the 5 strata, S1-5, including narrow thorny, large sparse, large moderate, large dense, large radiate, narrow wavy, large very sparse, and fine very sparse. There were four types of broadly stratified cells with dendritic trees extending from S4 to S2. One type resembled the parvocellular giant cell and another the broad thorny type described previously in primates. Another broadly stratified cell was called multi-tufted based on its distinctive dendritic branching pattern. The fourth type had been described previously, but not named; we called it broad wavy. There was a bistratified type with its major arbor in S5, the same level as the blue cone bipolar cell; it resembled the large, bistratified cell with blue ON-yellow OFF responses described recently. Two wide-field ganglion cell types were classified as diffuse because they had dendrites throughout the IPL. One had many small branches and was named thorny diffuse. The second was named smooth diffuse because it had straighter dendrites that lacked these processes. Dendrites of the large moderate and multi-tufted cells cofasciculated with ON starburst cell dendrites and were, therefore, candidates to be ON- and ON-OFF direction-selective ganglion cells, respectively. We concluded that there are at least 15 morphoplogical types of wide-field ganglion cells in macaque retinas. PMID- 16212698 TI - Mosaic properties of midget and parasol ganglion cells in the marmoset retina. AB - We measured mosaic properties of midget and parasol ganglion cells in the retina of a New World monkey, the common marmoset Callithrix jacchus . We addressed the functional specialization of these populations for color and spatial vision, by comparing the mosaic of ganglion cells in dichromatic ("red-green color blind") and trichromatic marmosets. Ganglion cells were labelled by photolytic amplification of retrograde marker ("photofilling") following injections into the lateral geniculate nucleus, or by intracellular injection in an in vitro retinal preparation. The dendritic-field size, shape, and overlap of neighboring cells were measured. We show that in marmosets, both midget and parasol cells exhibit a radial bias, so that the long axis of the dendritic field points towards the fovea. The radial bias is similar for parasol cells and midget cells, despite the fact that midget cell dendritic fields are more elongated than are those of parasol cells. The dendritic fields of midget ganglion cells from the same (ON or OFF) response-type array show very little overlap, consistent with the low coverage of the midget mosaic in humans. No large differences in radial bias, or overlap, were seen on comparing retinae from dichromatic and trichromatic animals. These data suggest that radial bias in ganglion cell populations is a consistent feature of the primate retina. Furthermore, they suggest that the mosaic properties of the midget cell population are associated with high spatial resolution rather than being specifically associated with trichromatic color vision. PMID- 16212699 TI - Horizontal cell morphology in nocturnal and diurnal primates: a comparison between owl-monkey (Aotus) and capuchin monkey (Cebus). AB - Horizontal cell morphology was studied in the retina of the nocturnal owl-monkey, Aotus, and compared with that of its diurnal, close relative, the capuchin monkey, Cebus . Cells were initially labeled with DiI and the staining was later photoconverted in a stable precipitated using DAB as chromogen. The sizes of cell bodies, dendritic fields, and axon terminals, number of dendritic clusters, intercluster spacing, and intercone spacing were measured at increasing eccentricities. Two distinct morphological classes of horizontal cells were identified, which resembled those of H1 and H3 cells described in diurnal monkeys. A few examples of a third class, possibly corresponding to the H2 cells of diurnal monkeys, were labeled. Both H1 and H3 cells increased in size and had increasing numbers of dendritic clusters with eccentricity. H3 cells were larger and had a larger number of dendritic clusters than H1 cells. Owl-monkey H1 cells had larger dendritic fields than capuchin monkey H1 cells at all quadrants in the central and midperipheral retinal regions, but the difference disappeared in the far periphery. Owl-monkey and capuchin monkey H1 cells had about the same number of dendritic clusters across eccentricity. As owl-monkey H1 cells were larger than capuchin monkey H1 cells, the equal number of clusters in these two primates was due to the fact that they were more spaced in the owl-monkey cells. H1 intercluster distance closely matched intercone spacing for both the owl-monkey and capuchin monkey retinas. On the other hand, H3 intercluster distance was larger than intercone spacing in the retina of both primates. Owl-monkey H1 axon terminals had 2-3 times more knobs than capuchin monkey H1 axon terminals in spite of having about the same size and, consequently, knob density was 2-3 times higher for owl-monkey than capuchin monkey H1 axon terminals across all eccentricities. The differences observed between owl-monkey and capuchin monkey horizontal cells, regarding the morphology of their dendritic trees and axon terminals, may be related to the differences found in the cone-to-rod ratio in the retina of these two primates. They seem to represent retinal specializations to the nocturnal and diurnal life styles of the owl-monkey and capuchin monkey, respectively. PMID- 16212701 TI - Response of the difference-of-Gaussians model to circular drifting-grating patches. AB - Forty years ago R.W. Rodieck introduced the Difference-of-Gaussians (DOG) model, and this model has been widely used by the visual neuroscience community to quantitatively account for spatial response properties of cells in the retina and lateral geniculate nucleus following visual stimulation. Circular patches of drifting gratings are now regularly used as visual stimuli when probing the early visual system, but for this stimulus type the mathematical evaluation of the DOG model response is significantly more complicated than for moving bars, full-field drifting gratings, or circular flashing spots. Here we derive mathematical formulas for the DOG-model response to centered circular patch gratings. The response is found to be given as the difference between two summed series, where each term in the series involves the confluent hypergeometric function. This function is available in commonly used mathematical software, and the results should thus be readily applicable. Example results illustrate how a strong surround suppression in area-summation curves for iso-luminant circular spots may be reversed into a surround enhancement for circular patch gratings. They also show that the spatial-frequency response changes from band-pass to low-pass when going from the full-field grating situation to the situation where the patch covers only the receptive-field center. PMID- 16212700 TI - Toward a unified model of vertebrate rod phototransduction. AB - Recently, we introduced a phototransduction model that was able to account for the reproducibility of vertebrate rod single-photon responses (SPRs) (Hamer et al., 2003). The model was able to reproduce SPR statistics by means of stochastic activation and inactivation of rhodopsin (R*), transducin (G alpha ), and phosphodiesterase (PDE). The features needed to capture the SPR statistics were (1) multiple steps of R* inactivation by means of multiple phosphorylations (followed by arrestin capping) and (2) phosphorylation dependence of the affinity between R* and the three molecules competing to bind with R* (G alpha, arrestin, and rhodopsin kinase). The model was also able to account for several other rod response features in the dim-flash regime, including SPRs obtained from rods in which various elements of the cascade have been genetically disabled or disrupted. However, the model was not tested under high light-level conditions. We sought to evaluate the extent to which the multiple phosphorylation model could simultaneously account for single-photon response behavior, as well as responses to high light levels causing complete response saturation and/or significant light adaptation (LA). To date no single model, with one set of parameters, has been able to do this. Dim-flash responses and statistics were simulated using a hybrid stochastic/deterministic model and Monte-Carlo methods as in Hamer et al. (2003). A dark-adapted flash series, and stimulus paradigms from the literature eliciting various degrees of light adaptation (LA), were simulated using a full differential equation version of the model that included the addition of Ca2+-feedback onto rhodopsin kinase via recoverin. With this model, using a single set of parameters, we attempted to account for (1) SPR waveforms and statistics (as in Hamer et al., 2003); (2) a full dark-adapted flash-response series, from dim flash to saturating, bright flash levels, from a toad rod; (3) steady-state LA responses, including LA circulating current (as in Koutalos et al., 1995) and LA flash sensitivity measured in rods from four species; (4) step responses from newt rods ( Forti et al., 1989) over a large dynamic range; (5) dynamic LA responses, such as the step-flash paradigm of Fain et al. (1989), and the two-flash paradigm of Murnick and Lamb (1996); and (6) the salient response features from four knockout rod preparations. The model was able to meet this stringent test, accounting for almost all the salient qualitative, and many quantitative features, of the responses across this broad array of stimulus conditions, including SPR reproducibility. The model promises to be useful in testing hypotheses regarding both normal and abnormal photoreceptor function, and is a good starting point for development of a full-range model of cone phototransduction. Informative limitations of the model are also discussed. PMID- 16212702 TI - Gradients of cone differentiation and FGF expression during development of the foveal depression in macaque retina. AB - Cones in the foveola of adult primate retina are narrower and more elongated than cones on the foveal rim, which in turn, are narrower and more elongated than those located more eccentric. This gradient of cone morphology is directly correlated with cone density and acuity. Here we investigate the hypothesis that fibroblast growth factor (FGF) signaling mediates the morphological differentiation of foveal cones--in particular, the mechanism regulating the elongation of foveal cones. We used immunoreactivity to FGF receptor (R) 4, and quantitative analysis to study cone elongation on the horizontal meridian of macaque retinae, aged between foetal day (Fd) 95 and 2.5 years postnatal (P 2.5 y). We also used in situ hybridization and immunohistochemistry to investigate the expression patterns of FGF2 and FGFR1-4 at the developing fovea, and three other sample locations on the horizontal meridian. Labeled RNA was detected using the fluorescent marker "Fast Red" (Roche) and levels of expression in cone inner segments and in the ganglion cell layer (GCL) were compared using confocal microscopy, optical densitometry, and tested for statistical significance. Our results show that morphological differentiation of cones begins near the optic disc around Fd 95, progressing toward the developing fovea up until birth, approximately. Levels of FGF2 and FGFR4 mRNAs expression are low in foveal cones, compared with cones closer to the optic disc, during this period. There is no similar gradient of FGF2 mRNA expression in the ganglion cell layer of the same sections. Maturation of foveal cones is delayed until the postnatal period. The results suggest that a wave of cone differentiation spreads from the disc region toward the developing fovea during the second half of gestation in the macaque. A gradient of expression of FGFR4 and FGF2 associated with the wave of differentiation suggests that FGF signalling mediates cone narrowing and elongation. PMID- 16212703 TI - Regularity and packing of the horizontal cell mosaic in different strains of mice. AB - The present study describes the relationships between mosaic regularity, intercellular spacing, and packing of horizontal cells across a two-fold variation in horizontal cell density in four strains of mice. We have tested the prediction that mosaic patterning is held constant across variation in density following our recent demonstration that intercellular spacing declines as density increases, by further examination of that dataset: Nearest-neighbor and Voronoi domain analyses were conducted on multiple fields of horizontal cells from each strain, from which their respective regularity indices were calculated. Autocorrelation analysis was performed on each field, from which the density recovery profile was generated, and effective radius and packing factor were calculated. The regularity indexes showed negative correlations with density rather than being held constant, suggesting that the strong negative correlation between intercellular spacing and density exceeded that required to produce a simple scaling of the mosaic. This was confirmed by the negative correlation between packing factor and density. These results demonstrate that the variation in the patterning present in the population of horizontal cells across these strains is a consequence of epigenetic mechanisms controlling intercellular spacing as a function of density. PMID- 16212704 TI - Localization of ionotropic glutamate receptors to invaginating dendrites at the cone synapse in primate retina. AB - The separation of OFF pathways that signal light decrements from ON pathways that signal light increments occurs at the first retinal synapse. The dendrites of OFF bipolar cells abut the cone pedicle at basal positions distal to the site of glutamate release and express ligand-gated or ionotropic glutamate receptors (GluR). The dendrites of ON bipolar cells penetrate narrow invaginations of the cone pedicle proximal to the site of release and express the G-protein-coupled, metabotropic glutamate receptor, mGluR6. However, recent studies demonstrating the expression of GluR subunits in the rodent rod bipolar cell, known to yield an ON response to light, call this basic segregation of receptors into question. The light-microscopic distribution of many glutamate receptors in the primate retina is now well established. We reexamined their ultrastructural localization in the outer retina of Macaca fascicularis to test systematically whether invaginating dendrites at the cone synapse, presumably from ON bipolar cells, also express one or more ionotropic subunits. Using preembedding immunocytochemistry for electron microscopy, we quantified the distribution of the AMPA-sensitive subunits GluR2/3 and GluR4 and of the kainate-sensitive subunits GluR6/7 across 207 labeled dendrites occupying specific morphological loci at the cone pedicle. We report, in agreement with published investigations, that the majority of labeled processes for GluR2/3 (70%) and GluR4 (67%) either occupy basal positions or arise from horizontal cells. For GluR6/7, we find a significantly lower fraction of labeled processes at these positions (47%). We also find a considerable number of labeled dendrites for GluR2/3 (10%), GluR4 (21%), and GluR6/7 (18%) at invaginating positions. Surprisingly, for each subunit, the remainder of labeled processes corresponds to "fingers" of presynaptic cytoplasm within the cone invagination. PMID- 16212705 TI - Spatial coding and response redundancy in parallel visual pathways of the marmoset Callithrix jacchus. AB - Many neurons in the primary visual cortex (area V1) show pronounced selectivity for the orientation and spatial frequency of visual stimuli, whereas most neurons in subcortical afferent streams show little selectivity for these stimulus attributes. It has been suggested that this transformation is a functional sign of increased coding efficiency, whereby the redundancy (or overlap in response properties) is reduced at consecutive levels of visual processing. Here we compared experimentally the response redundancy in area V1 with that in the three main dorsal thalamic afferent streams, the parvocellular (PC), koniocellular (KC), and magnocellular (MC) divisions of the dorsal lateral geniculate nucleus (LGN) in marmosets. The spatial frequency and orientation tuning of single cells in the LGN and area V1 were measured, using luminance contrast sine-wave gratings. A joint spatial frequency-orientation response selectivity profile was calculated for each cell. Response redundancy for each population was estimated by cross-multiplication of the joint selectivity profiles for pairs of cells. We show that when estimated in this way, redundancy in LGN neurons is approximately double that of neurons in cortical area V1. However, there are differences between LGN subdivisions, such that the KC pathway has a spatial representation that lies between the redundant code of the PC and MC pathways and the more efficient sparse spatial code of area V1. PMID- 16212707 TI - Photoreceptors in the rat retina are specifically vulnerable to both hypoxia and hyperoxia. AB - The current study aims to assess the vulnerability of photoreceptors in rat retina to variations in tissue oxygen levels. Young adult Sprague-Dawley rats were exposed to air with the concentration of oxygen set at 10% (hypoxia), 21% (room air, normoxia), and four levels of hyperoxia (45%, 65%, 70%, and 75%), for up to 3 weeks. Their retinas were then examined for cell death, using the TUNEL technique. Hypoxia (10% oxygen) for 2 weeks caused a limited but significant rise in the frequency of TUNEL+ (dying) cells in the retina, the great majority (>90%) being located in the outer nuclear layer (ONL). Hyperoxia also induced an increase in the frequency of TUNEL+ cells, again predominantly in the ONL. The increase rose with duration of exposure, up to 2 weeks. At 2 weeks exposure, the increase was limited yet significant at 45% oxygen, and maximal at 65%. Where the frequencies of TUNEL+ cells were high, it was evident that photoreceptor death was maximal in the midperipheral retina. The adult retina is vulnerable to maintained shifts in oxygen availability to the retina, both below and above normal. The vulnerability is specific to photoreceptors; other retinal neurons appeared resistant to the exposures tested. Shifts in retinal oxygen levels caused by variations in ambient light, by the persistence of light through the normally dark (night) half of the day-night cycle, or by depletion of the photoreceptor population, may contribute to photoreceptor death in the normal retina. PMID- 16212706 TI - Spatiotemporal integration of light by the cat X-cell center under photopic and scotopic conditions. AB - Visual responses to stimulation at high temporal frequency are generally considered to result from signals that avoid light adaptive gain adjustment, simply reflecting linear summation of luminance. Under conditions of high photopic illuminance, the center of the receptive field of the cat X-cell has been shown to expand in size when stimulated at high temporal frequency, raising the possibility that there is spatiotemporal interaction in luminance summation. Here we show that this expansion maintains constant the product of the center's luminance summing area and the temporal period of luminance modulation, implying that spatial and temporal integration of luminance can be traded for one another by the X-cell center. As such the X-cell has a spatiotemporal window for luminance integration that fuses the classical concepts of a spatial window of luminance integration (Ricco's Law) with a temporal window of luminance integration (Bloch's Law). We were interested to determine whether this tradeoff between spatial and temporal summation of luminance occurs also at lower light levels, where the temporal-frequency bandwidth of the X-cell is narrower. We found that it does not. Center radius does not expand with temporal frequency under either low photopic or scotopic conditions. These results are discussed within the context of the known retinal circuitry that underlies the X-cell center for photopic and scotopic conditions. PMID- 16212708 TI - "Small-tufted" ganglion cells and two visual systems for the detection of object motion in rabbit retina. AB - Small-tufted (ST) ganglion cells of rabbit retina are divided into eight types based upon morphology, branching pattern, level of dendritic stratification, and quantitative dimensional analysis. Only one of these types has been previously characterized in Golgi preparations, and four may be discerned in the work of others. Given their small dendritic-field size, and assuming uniform mosaics of each across the retina, ST cells comprise about 45% of all rabbit ganglion cells, and are therefore of major functional significance. Four ST cells occur as two paramorphic (a/b) pairs, and thus belong to class III, as previously defined. Four branch in sublaminae a and b of the inner plexiform layer (IPL) and therefore belong to class IV. ST cells have small cell bodies 10-15 microm in diameter, small axons 0.7-1.3 microm in diameter, and small dendritic-field diameters, 40-110 microm in mid-visual streak. The dendrites of ST cells are highly branched, and bear spines and appendages of varying length, but vary from type to type. Class III.2 cells and class III.3 cells are partly bistratified. Class IV small-tufted cells differ characteristically in multiple features of dendritic branching and stratification. Class III small-tufted cells apparently have concentric (ON-center and OFF-center) receptive fields and may have "sluggish-transient" (class III.2) and "sluggish-sustained" (class III.3) physiology. Class IV cells include the "local-edge-detector" (LED) (class IVst1), and are all expected to give ON-OFF responses to small, centered, slowly moving visual stimuli. Based upon systematic variation in dendritic-field size across the retina, ST cells may be divided into two groups. In this "universal prey" species, they may belong to two systems of motion detection, typified by ON-OFF directionally selective and LED ganglion cells, respectively, specialized for detection of rapid motion at the horizon for land-based predators, and slow motion for airborne predators. PMID- 16212710 TI - A review of controlled trials in the pharmacological treatment of premature ejaculation. AB - Premature ejaculation is a common sexual problem which presents to genitourinary (GU) medicine services. Five main treatment approaches have been used in clinical trials: behavioural therapy, antidepressants, phosphodiesterase-5 (PDE5) inhibitors, topical anaesthetic agents and alpha-blockers. We have carried out a systematic review of published pharmacological trials. All antidepressants appeared to delay ejaculation to some extent at all doses. Anaesthetic creams appeared to be as successful in slowing ejaculation as antidepressants without systemic side-effects, although some patients did experience erectile problems or unpleasant local symptoms. Anecdotally, behavioural therapy is effective and appears to have long-lasting efficacy. There is a need for quality comparative trial of behavioural therapy, topical anaesthetic agents and antidepressants, including appropriate measures of relapse, follow-up and acceptability of continuing long-term treatment. PMID- 16212711 TI - The psychosexual and psychosocial profile of male genitourinary medicine patients with a history of sexual abuse/assault or unwanted sexual experiences. AB - One hundred and forty-one male patients who attended a District General Hospital's department of genitourinary medicine completed a questionnaire including demographic indices, history of sexual abuse/assault, anxiety and depression, and sexual concerns and attitudes. Of these, 52 (36.9%) reported past unwanted sexual experiences (USE). A significantly higher proportion of the USE group consisted of homosexual men than the non-USE majority. The prevalence rate for USE involving other men was 15.6%, which is comparable to previous studies. Hospital Anxiety and Depression scores were significantly greater for patients with a history of USE (t = -2.82, P = 0.006, for anxiety and t = -3.01, P = 0.003 for depression). Patients with a history of USE also scored significantly higher on a measure of sexual concerns. However, there were no significant differences between the USE and non-USE groups on two brief measures of alcohol and drug consumption. Clinicians in sexual health settings need to be sensitive to the fact that a significant proportion of male patients may have an undisclosed history of USE. PMID- 16212709 TI - Stratification of alpha ganglion cells and ON/OFF directionally selective ganglion cells in the rabbit retina. AB - The correlation between cholinergic sensitivity and the level of stratification for ganglion cells was examined in the rabbit retina. As examples, we have used ON or OFF alpha ganglion cells and ON/OFF directionally selective (DS) ganglion cells. Nicotine, a cholinergic agonist, depolarized ON/OFF DS ganglion cells and greatly enhanced their firing rates but it had modest excitatory effects on ON or OFF alpha ganglion cells. As previously reported, we conclude that DS ganglion cells are the most sensitive to cholinergic drugs. Confocal imaging showed that ON/OFF DS ganglion cells ramify precisely at the level of the cholinergic amacrine cell dendrites, and co-fasciculate with the cholinergic matrix of starburst amacrine cells. However, neither ON or OFF alpha ganglion cells have more than a chance association with the cholinergic matrix. Z -axis reconstruction showed that OFF alpha ganglion cells stratify just below the cholinergic band in sublamina a while ON alpha ganglion cells stratify just below cholinergic b . The latter is at the same level as the terminals of calbindin bipolar cells. Thus, the calbindin bipolar cell appears to be a prime candidate to provide the bipolar cell input to ON alpha ganglion cells in the rabbit retina. We conclude that the precise level of stratification is correlated with the strength of cholinergic input. Alpha ganglion cells receive a weak cholinergic input and they are narrowly stratified just below the cholinergic bands. PMID- 16212712 TI - The role of illness perceptions: psychological distress and treatment-seeking delay in patients with genital warts. AB - This study examined the relationships between illness perceptions, psychological distress and treatment-seeking delay in genital warts patients. Sixty-six genital warts patients were approached while attending a sexual health clinic. They completed a questionnaire assessing their illness perceptions, psychological distress and treatment-seeking delay. Negative perceptions of illness consequences and control and a perceived cyclical timeline were associated with increased psychological distress. Perceived illness consequences maintained significance in a multiple regression equation, which accounted for 25% variance in distress. Depression was associated with treatment-seeking delay (r = 0.28, P = 0.03). In conclusion, illness perceptions may play an important role in the experience of psychological distress in genital warts patients. The implications of these findings for the design of health-care interventions are discussed. PMID- 16212713 TI - The risk of needle stick accidents during surgical procedures: HIV-1 viral load in blood and bone marrow. AB - Health-care workers are at risk to acquire HIV through occupational exposure to blood of HIV-infected patients. The mean risk after a percutaneous exposure is approximately 0.3%. A large inoculum and a source patient with a high plasma viral load increases the transmission risk. To ensure the safety of the operating team, we try to reduce HIV viral load in plasma prior to high-risk interventions (cardiothoracic and orthopaedic surgery). However, in 15.7% of the exposures occurring in the operating room, the possible source material is bone marrow. To make more accurate exposure risk assessments, we measured HIV-1 RNA in both plasma and bone marrow of five HIV-infected patients undergoing surgery. We found that the plasma viral load was not different from the viral load in bone marrow. PMID- 16212714 TI - Herpes zoster in HIV-1-infected patients in the era of highly active antiretroviral therapy: a prospective observational study. AB - Between June 1994 and May 2003, 93 of 716 (13.0%) HIV-infected patients with a median baseline cell differentiation CD4+ count of 61 x 10(6) cells/L (range, 1 1206 x 10(6) cells/L) developed 103 episodes of herpes zoster [HZ], with an incidence of 5.67 per 100 person-years (PY). The incidence of HZ in the pre highly active antiretroviral therapy (HAART) era (17.21 per 100 PY) was significantly higher than that in the post-HAART era (5.05 per 100 PY) (P < 0.0001). In the first six months of enrollment, the incidence of HZ was significantly higher than that between six and 12 months both in the pre-HAART (27.65 per 100 PY versus 8.43 per 100 PY, P = 0.02) and post-HAART era (17.79 per 100 PY versus 3.39 per 100 PY, P < 0.0001). In multivariate analyses, only baseline CD4+ count remained a significant risk factor associated with HZ. HZ did not increase mortality rate either in the pre-HAART or post-HAART era, although the risk for HIV progression was significantly higher in patients with HZ (adjusted odds ratio [OR], 1.747, 95% confidence interval, 1.037-2.943). We conclude that the incidence of HZ was highest in the first six months of enrollment in patients at late stage of HIV infection, which did not increase with the introduction of HAART. Baseline CD4+ lymphocyte count was the most significant risk factor associated with development of HZ. HZ was associated with increased risk for HIV progression, but not mortality. PMID- 16212715 TI - Atherogenic lipid profile and cardiovascular risk factors in HIV-infected patients (Netar Study). AB - We undertook a transverse study of 603 HIV outpatients to determine their atherogenic lipid profile (ALP) and cardiovascular risk (CVR) factors. CVR was estimated from the Framingham score. ALP was defined as a total cholesterol to high density lipoprotein (HDL)-cholesterol ratio > or =5 plus triglycerides > or =150 mg/dL and a CVR >10% at 10 years was considered high. The most frequent CVR factor was smoking. ALP was diagnosed in 26.9% and was related to sex (odds ratio [OR] 2.6; 95% confidence interval [CI], 1.3-5.0; P = 0.0047), protease inhibitor use (OR 3.8; 95% CI, 1.8-7.8; P = 0.0002) and sexual HIV risk (OR 2.4; 95% CI, 1.4-4.0; P = 0.0004). The mean 10-year CVR was 6.2%, was high in 20.4% and was related to sexual HIV-risk (OR 3.8; 95% CI, 2.1-6.8; P < 0.00001) and nadir cell differentiation factor (CD4) (OR 1.0; 95% CI, 1.0-1.003; P = 0.0026). Although the current CVR of our patients is not high, the contribution to the lipid profile of highly active antiretroviral therapy (HAART)-associated factors and the high prevalence of some risk factors may lead to an increased future CVR. PMID- 16212716 TI - Are women with chlamydia infection who self-refer to genitourinary medicine clinics different? AB - The objective of this study was to explore whether patients with Chlamydia trachomatis infection who self-refer to genitourinary medicine clinics have different demographic characteristics to those who initially attend other agencies. This study took place in three genitourinary medicine clinics from Birmingham, Nottingham and Sheffield. Demographic and post-code data were collected from female patients diagnosed with genital chlamydia infection in 2000. Townsend scores, as an index of socioeconomic status, were derived from post-codes from a subset of the cohort (from Birmingham). Comparison was made between those who were diagnosed by genitourinary medicine clinics and those diagnosed in the community and referred to genitourinary medicine clinics for further management. Data were collected from 1047 genitourinary medicine and 816 non-genitourinary medicine women, of whom 686 (84.1%) attended genitourinary medicine clinics following referral. After excluding those with incomplete data, 1614 (987 genitourinary medicine and 627 non-genitourinary medicine) patients were included in the study. Using logistic regression analysis, we were unable to demonstrate any significant differences in age or Townsend scores between genitourinary medicine and non-genitourinary medicine patients. However, significantly more Black Caribbean (odds ratio [OR] = 2.72, 95% confidence interval [CI]: 2.22, 3.20) and single women (OR = 1.97, 95% CI: 1.64, 2.29) self referred to genitourinary medicine clinics compared with other health-care settings. This trend was consistent between Birmingham and Nottingham. In Sheffield, there was no difference in marital status. Ethnicity was not a factor as there were no Black Caribbean patients in the Sheffield cohort. Women who were diagnosed with genital chlamydia infection in genitourinary medicine clinics have some different demographic characteristics to those who were diagnosed in the community. PMID- 16212717 TI - Diagnostic utility of bone marrow sampling in HIV-infected patients since the advent of highly active antiretroviral therapy. AB - We investigated the diagnostic value of bone marrow (BM) sampling in investigation of HIV-infected patients presenting to a major London HIV treatment centre between 1999 and 2004. One hundred and fourteen consecutive patients underwent 130 BM samplings. The majority of BM aspirates were normal or showed non-diagnostic changes; microscopy revealed lymphoma in one and mycobacterial infection in two. Subsequent culture identified mycobacterial infection in nine samples. BM trephine had a diagnostic yield of 26% in patients with fever and cytopaenia (including mycobacteriosis in 14%, lymphoma in 6%, Castleman disease in 3% and "drug effect" in 3%), a yield of 20% in patients with fever, but no cytopaenia (mycobacteriosis in each case), and a yield of 19% in patients with cytopaenia in the absence of fever (lymphoma in 5% and "drug effect" in 14%). In investigation/staging of lymphoma, the diagnostic yield was 36%. The overall yield from BM sampling was 30% in patients receiving highly active antiretroviral therapy (HAART) and 23% in those not receiving HAART. In this study, BM sampling was of most diagnostic value in HIV-infected patients where fever and cytopaenia coexisted in the absence of localizing signs of infection, and in the staging/investigation of lymphoma. BM sampling had less diagnostic value in the investigation of fever without cytopaenia or cytopaenia without fever. PMID- 16212718 TI - Sociodemographic and clinical characteristics of clients presenting for HIV voluntary counselling and testing in Moshi, Tanzania. AB - HIV voluntary counselling and testing (VCT) reduces high-risk sexual behaviour. Factors associated with HIV infection in VCT clients have not been well characterized in northern Tanzania. We prospectively surveyed 813 VCT clients in Moshi, Tanzania. Clients were administered a questionnaire on sociodemographic characteristics, sexual behaviour, and health status. Blood was taken for rapid HIV antibody testing. Factors associated with HIV seropositivity were identified using multivariate logistic regression analysis. Of 813 clients, the seroprevalence was 16.7%. The strongest associations with seropositivity were reporting diarrhoea (odds ratio [OR] 10.4, 95% confidence interval [CI] 3.6 29.9), an ill sexual partner (OR 6.3, 95% CI 3.0-12.9), or being a woman (OR 3.5, 95% CI 2.0-6.3). In a separate regression, the number of symptoms also predicted HIV infection (OR 2.1, 95% CI 1.6-2.6). VCT clients who tested positive had more HIV-related symptoms suggesting presentation at a later stage of HIV infection. PMID- 16212719 TI - Clinicians' perspective of a training programme in syndromic management of sexually transmitted infections in Northern Tanzania. AB - This process evaluation study was designed to investigate clinicians' perception of a training programme on syndromic management of sexually transmitted infections (STIs) and their experiences in applying the skills learnt during the course. Out of 136 eligible course participants from Arusha and Kilimanjaro regions, Tanzania, 47 were invited to take part in the study and 40 agreed to participate (85%). The research instruments consisted of a structured interview including open- and close-ended questions, a self-administered questionnaire, and an observation checklist. While the clinicians reported to be satisfied with the course itself, the results revealed insufficient practical training. The clinicians reported problems with condom promotion and partner notification, and frequent shortage of equipment and drugs in the clinics was observed. Future courses on syndromic management of STI should focus on skills training related to health education and counselling, and have a stronger emphasis on practical, clinical work. There is need to address barriers to condom promotion and partner referral. PMID- 16212720 TI - Patients unaware of their HIV infection until AIDS diagnosis in Sweden 1996-2002- a remaining problem in the highly active antiretroviral therapy era. AB - Our objective was to analyse the characteristics of patients who were unaware of their HIV infection until they developed AIDS, in the period after introduction of highly active antiretroviral therapy. The complete national register of HIV and AIDS cases reported to the Department of Epidemiology at the Swedish Institute for Infectious Disease Control 1996-2002 was searched for cases diagnosed with HIV less than three months before AIDS diagnosis (so-called "late testers"). Of a total of 487 patients with AIDS, reported during the seven-year period, 219 (45%) were late testers. Their proportion of all AIDS cases increased from 22% in 1996 to 58% in 2002. Heterosexual route of transmission, age greater than 40 years, and foreign origin were all significant risk factors for being a late tester. Intravenous drug users were associated with a highly significant reduced risk. The group without previously known HIV infection represents an increasing part of all cases of AIDS. From a disease control and from a medical perspective, it is important to study this group further and discover what measures are needed for earlier identification and access to medical care. PMID- 16212721 TI - Knowledge of human papillomavirus and effects on sexual behaviour of gay/bisexual men: a brief report. AB - The objective of this study was to examine the knowledge and misinformation about human papillomavirus (HPV) and differences in sexual risk practices. Self-report surveys assessing the history of HPV/genital warts and sexual practices were completed by 1065 gay/bisexual men in New York City. Of the men reporting a history of HPV, genital warts, or both, the majority reported having warts but not HPV, demonstrating inaccurate knowledge. A significantly greater percentage of men who reported both HPV and warts (HPV+) reported having unsafe sex compared with men reporting neither (HPV-) and men reporting warts but not HPV (HPV+ uninformed). HPV+ and HPV+ uninformed men reported significantly more men non primary sex partners in the past three months than HPV- men. Findings indicate that many gay/bisexual men, a population at risk for HPV, are misinformed about its various permutations. Men who are HPV+ report increased sexual risk practices and more sexual partners. Comprehensive gay men's health programmes must include HPV education. PMID- 16212722 TI - Pharmacological treatment for premature ejaculation. AB - Premature ejaculation is a common male sexual dysfunction. Treatment modalities as recommended by the British Association of Sexual Health and HIV include behavioural therapy, tricyclic antidepressants, selective serotonin reuptake inhibitors (SSRIs) and local anaesthetic creams. We audited the clinical cohort from our dedicated sexual dysfunction clinic to determine the success of prescribed treatment and co-existing prostatitis/male pelvic pain, erectile dysfunction, phosphodiesterase-5 (PDE5) inhibitor use and anxiety. The use of SSRIs was successful in the treatment of premature ejaculation with or without the use of local anaesthetic cream. Co-existing prostatitis/male pelvic pain, erectile dysfunction, PDE5 inhibitor use and anxiety were high. PMID- 16212723 TI - A multi-district audit of the management of HIV infection in pregnant women and the prevention of mother-to-child transmission: Yorkshire HIV pregnancy audit. AB - A regional audit was performed to compare clinical practice against the British HIV Association guidelines for the management of HIV infection in pregnancy. Data were collected from 2000 to 2002 from eight clinics across Yorkshire using a questionnaire. There were 22 live births to 22 HIV + pregnant women. There were no cases of transmission of HIV from mother to child. The majority (20/22) of mothers received therapy as recommended in the guidelines, with only two initially receiving inappropriate dual therapy. In all, 16/22 (73%) had elective caesarean sections, 5/22 (23%) emergency sections after the onset of labour and one had a vaginal delivery. Also, 12/22 (55%) definitely received intravenous zidovudine during delivery. Added to these, 19/22 infants received appropriate antiretroviral medication for four to six weeks after birth. No mothers were known to have breast-fed. PMID- 16212725 TI - Making health policy: networks in research and policy after 1945. AB - Science and policy in health and medicine have interacted in new ways in Britain since 1945. The relationship between research and policy has a history. The changing role of social medicine, the rise of health services research and "customer contractor" policies in government have been important. The relationship between research and policy has been analysed by different schools of thought. This chapter categorises them as several groups: "evidence-based", "journalism", "sociology of scientific knowledge" and "science policy studies". The chapters in the book illuminate aspects of these changing relationships. The role of chronic disease epidemiology, of new networks in public health, of media focussed activism, and of health technology and its advocates have been more important than political interest. PMID- 16212726 TI - "Association or causation?" The debate on the scientific status of risk factor epidemiology, 1947-c. 1965. AB - In the second half of the twentieth century, epidemiology came to shape public health discourses and practices to an unprecedented extent. The chapter explores the transformation of the discipline after World War Two and analyses the crucial debate on the notion of "causation" that sprung from the growing interest in non transmissible, chronic diseases. A landmark in this history was the controversy over the interpretation of the statistical relationship between smoking and lung cancer prompted by American and British publications in 1950. This sometimes heated debate also provided Austin Bradford Hill with the opportunity to set out his "pragmatics" of risk factor epidemiology. PMID- 16212727 TI - Who makes alcohol policy? Science and policy networks 1950-2000. AB - This chapter examines the interaction between alcohol policy networks, ideological "frames" of understanding of the alcohol problem and the production and use of research evidence. The emergence and growth of policy networks and the production of research based "evidence" as the rationale for change is traced from the mid-nineteenth century over major shifts in understanding of the problem, from a "moral" model to a "disease" model to a "public health" approach and, finally, to an approach which brings together health and criminal justice perspectives. The chapter challenges policy discourse which presents a rational model of policy making supported by "scientific" research. PMID- 16212728 TI - Issue network versus producer network? ASH, the Tobacco Products Research Trust and UK smoking policy. AB - Policy science studies of networks in smoking policy segment the smoking arena into a "producer network" of industrial and retail interests and an "issue network" of anti-smoking organisations. Case studies of ASH (Action on Smoking and Health) and the Tobacco Products Research Trust (TPRT) indicate that networks in smoking policy were more complex and overlapping. ASH pioneered a new style of media-conscious health activism in the 1970s with an anti-industry line. Nevertheless the strategy of harm reduction remained an objective for industry and government, and also for some public health interests through the work of the TPRT. First safer smoking and then nicotine were the focus of these activities. PMID- 16212729 TI - British expert advice on diet and heart disease C. 1945-2000. AB - This chapter looks at the evolution over fifty years of government recommendations for reducing the risk of coronary heart disease amongst the British population. It also explores how non-governmental organisations such as the BMA established their own nutritional advice giving mechanisms and tried to influence such recommendations--and how networks of scientists constituted advisory committees. Scientists tended to favour recommendations which would lead to interventions within the scope of their own disciplines. It was also the case that consensus was lacking for a long while; this was only arrived at during the end of the period under consideration. PMID- 16212730 TI - Peer pressure and imposed consensus: the making of the 1984 Guidelines of Good Clinical Practice in the Treatment of Drug Misuse. AB - The role of evidence and "expert" opinion in forming drug treatment policies is explored though the case of the first clinical guidelines on drug misuse (1984). Developed to secure the ascendancy of one particular treatment model and impose this on all doctors, they cited no supporting research evidence. The experience of an expert committee was deemed sufficient by many of those involved for determining "good practice". This chapter considers the motives and alliances of the different factions of the state and medical profession responsible for the guidelines and how each succeeded or failed in achieving their goals. PMID- 16212731 TI - Evidence, experts and committees: the shaping of hospital pharmacy policy in Great Britain 1948 to 1974. AB - This chapter illustrates the rise of research-based evidence by reference to two post-war enquiries into the hospital pharmaceutical service in Great Britain. The first, by a sub-committee of the Central Health Services Council, resulted in the Linstead report of 1955; the second, by a working party appointed by the Minister of Health, in the Noel Hall report of 1970. The former had little impact, whilst the latter was the catalyst for monumental change. This chapter explores the reasons why. It demonstrates that by the late 1960s greater use was being made of statistics and research-based evidence, and that the Noel Hall working party represented a new style of expert committee that emerged at this time. PMID- 16212732 TI - Renal dialysis: counting the cost versus counting the need. AB - The relatively "lean and mean" provision of renal dialysis in Britain is a notorious example of "covert rationing", apparently achieved by a lack of central policy. Then in its first experiment in "target-setting" in the NHS in 1984, central government used the profession's preferred measure of need, thus promoting expansion of renal services, at almost exactly the time when the "quality-adjusted life year" (QALY) developed by health economists indicated that renal dialysis scored low in cost-benefit terms. This chapter examines these conundrums in terms of centre-periphery relations, clinical autonomy versus collective direction, and the politics of competing ways of counting need and cost. PMID- 16212733 TI - Intensive care: measurement and audit in an expensive growth area of medicine. AB - Intensive care historically shared with renal dialysis a relative shortfall in the UK and widely (wildly?) divergent provision in the regions. However, research on measurement of need was very different, with a focus on clinical audit. Priority was given to assessing how much the performance of different units could be attributed to differing intakes of patients through complex measuring tools such as the APACHE II score. While policy bodies welcomed this approach, policy guidelines in the 1990s relied most heavily on dissemination of "best practice", in any case raising standards through emulation within the UK rather than raising funding to European levels. PMID- 16212734 TI - Publicity as policy: the changing role of press and public relations at the BMA, 1940s-80s. AB - Relationships between policy and publicity or public relations (PR) have been questioned since the emergence of professional public relations in the early twentieth century. In the field of health and medicine organised PR activity began to flourish in the decades following World War Two. Its presence became evident in government departments, in professional associations, voluntary bodies and campaigning groups. Increasingly, policy decisions had to be publicly performed through rituals like the press conference. This chapter documents the development of press and PR activity at the British Medical Association (BMA) from the 1950s to the 1980s. The BMA provides a well-documented case, which can be used to suggest broader shifts in the association between policy and publicity. PMID- 16212735 TI - Networks of mass communication: reporting science, health and medicine in the 1950s and '60s. AB - Policy analysts routinely identify the media as a key player in the post-war policy process. However, we know little about the way this role has developed over time and in relation to different fields of interest. This chapter focuses on the emergence of institutions, mechanisms and professional groups which have served to manage the flow of public communication in relation to science, health and medicine. These are closely connected areas of practice and knowledge production. However, when they are examined as media sources or areas of media interest, each has a different cargo of historical associations. PMID- 16212737 TI - Emblematic monsters: unnatural conceptions and deformed births in early modern Europe. PMID- 16212750 TI - New medical challenges during the Scottish enlightenment. PMID- 16212761 TI - The effect of NMDA receptors on gain modulation. AB - The ability of individual neurons to modulate the gain of their input-output function is important for information processing in the brain. In a recent study, shunting inhibition was found to modulate the gain of cerebellar granule cells subjected to simulated currents through AMPA receptor synapses. Here we investigate the effect on gain modulation resulting from adding the currents mediated by NMDA receptors to a compartmental model of the granule cell. With only AMPA receptors, the changes in gain induced by shunting inhibition decreased gradually with the average firing rate of the afferent mossy fibers. With NMDA receptors present, this decrease was more rapid, therefore narrowing the bandwidth of mossy fiber firing rates available for gain modulation. The deterioration of gain modulation was accompanied by a reduced variability of the input current and saturation of NMDA receptors. However, when the output of the granule cell was plotted as a function of the average input current instead of the input firing frequency, both models showed very similar response curves and comparable gain modulation. We conclude that NMDA receptors do not directly impair gain control by shunting inhibition, but the effective bandwidth decreases as a consequence of the increased total charge transfer. PMID- 16212762 TI - Oscillatory synchronization requires precise and balanced feedback inhibition in a model of the insect antennal lobe. AB - In the insect olfactory system, odor-evoked transient synchronization of antennal lobe (AL) projection neurons (PNs) is phase-locked to the oscillations of the local field potential. Sensory information is contained in the spatiotemporal synchronization pattern formed by the identities of the phase-locked PNs. This article investigates the role of feedback inhibition from the local neurons (LNs) in this coding. First, experimental biological results are reproduced with a reduced computational spiking neural network model of the AL. Second, the low complexity of the model leads to a mathematical analysis from which a lower bound on the phase-locking probability is derived. Parameters involved in the bound indicate that PN phase locking depends not only on the number of LN-evoked inhibitory postsynaptic potentials (IPSPs) previously received, but also on their temporal jitter. If the inhibition received by a PN at the current oscillatory cycle is both perfectly balanced (i.e., equal to the mean inhibitory drive) and precise (without any jitter), then the PN will be phase-locked at the next oscillatory cycle with probability one. PMID- 16212764 TI - Incremental online learning in high dimensions. AB - Locally weighted projection regression (LWPR) is a new algorithm for incremental nonlinear function approximation in high-dimensional spaces with redundant and irrelevant input dimensions. At its core, it employs nonparametric regression with locally linear models. In order to stay computationally efficient and numerically robust, each local model performs the regression analysis with a small number of univariate regressions in selected directions in input space in the spirit of partial least squares regression. We discuss when and how local learning techniques can successfully work in high-dimensional spaces and review the various techniques for local dimensionality reduction before finally deriving the LWPR algorithm. The properties of LWPR are that it (1) learns rapidly with second-order learning methods based on incremental training, (2) uses statistically sound stochastic leave-one-out cross validation for learning without the need to memorize training data, (3) adjusts its weighting kernels based on only local information in order to minimize the danger of negative interference of incremental learning, (4) has a computational complexity that is linear in the number of inputs, and (5) can deal with a large number of-possibly redundant-inputs, as shown in various empirical evaluations with up to 90 dimensional data sets. For a probabilistic interpretation, predictive variance and confidence intervals are derived. To our knowledge, LWPR is the first truly incremental spatially localized learning method that can successfully and efficiently operate in very high-dimensional spaces. PMID- 16212763 TI - Response properties of an integrate-and-fire model that receives subthreshold inputs. AB - A computational technique is described for calculation of the interspike interval and poststimulus time histograms for the responses of an integrate-and-fire model to arbitrary inputs. The effects of the model parameters on the response statistics were studied systematically. Specifically, the probability distribution of the membrane potential was calculated as a function of time, and the mean interspike interval and PST histogram were calculated for arbitrary inputs. For stationary inputs, the regularity of the output was studied in detail for various model parameters. For nonstationary inputs, the effects of the model parameters on the output synchronization index were explored. The results show that enhanced synchronization in response to low-frequency stimuli required a large number (n > 25) of weak inputs. Irregular responses and a linear input output rate relationship required strong (but subthreshold) inputs with a small time constant. A model cell with mixed-amplitude synaptic inputs can respond to stationary inputs irregularly and have enhanced synchronization to nonstationary inputs that are phase-locked to low-frequency inputs. Both of these response properties have been reported for some cells in the ventral cochlear nucleus in the auditory brainstem. PMID- 16212765 TI - On the nonlearnability of a single spiking neuron. AB - We study the computational complexity of training a single spiking neuron N with binary coded inputs and output that, in addition to adaptive weights and a threshold, has adjustable synaptic delays. A synchronization technique is introduced so that the results concerning the nonlearnability of spiking neurons with binary delays are generalized to arbitrary real-valued delays. In particular, the consistency problem for N with programmable weights, a threshold, and delays, and its approximation version are proven to be NP-complete. It follows that the spiking neurons with arbitrary synaptic delays are not properly PAC learnable and do not allow robust learning unless RP = NP. In addition, the representation problem for N, a question whether an n-variable Boolean function given in DNF (or as a disjunction of O(n) threshold gates) can be computed by a spiking neuron, is shown to be coNP-hard. PMID- 16212766 TI - A novel model-based hearing compensation design using a gradient-free optimization method. AB - We propose a novel model-based hearing compensation strategy and gradient-free optimization procedure for a learning-based hearing aid design. Motivated by physiological data and normal and impaired auditory nerve models, a hearing compensation strategy is cast as a neural coding problem, and a Neurocompensator is designed to compensate for the hearing loss and enhance the speech. With the goal of learning the Neurocompensator parameters, we use a gradient-free optimization procedure, an improved version of the ALOPEX that we have developed, to learn the unknown parameters of the Neurocompensator. We present our methodology, learning procedure, and experimental results in detail; discussion is also given regarding the unsupervised learning and optimization methods. PMID- 16212767 TI - A robust information clustering algorithm. AB - We focus on the scenario of robust information clustering (RIC) based on the minimax optimization of mutual information (MI). The minimization of MI leads to the standard mass-constrained deterministic annealing clustering, which is an empirical risk-minimization algorithm. The maximization of MI works out an upper bound of the empirical risk via the identification of outliers (noisy data points). Furthermore, we estimate the real risk VC-bound and determine an optimal cluster number of the RIC based on the structural risk-minimization principle. One of the main advantages of the minimax optimization of MI is that it is a nonparametric approach, which identifies the outliers through the robust density estimate and forms a simple data clustering algorithm based on the square error of the Euclidean distance. PMID- 16212768 TI - Learning curves for stochastic gradient descent in linear feedforward networks. AB - Gradient-following learning methods can encounter problems of implementation in many applications, and stochastic variants are sometimes used to overcome these difficulties. We analyze three online training methods used with a linear perceptron: direct gradient descent, node perturbation, and weight perturbation. Learning speed is defined as the rate of exponential decay in the learning curves. When the scalar parameter that controls the size of weight updates is chosen to maximize learning speed, node perturbation is slower than direct gradient descent by a factor equal to the number of output units; weight perturbation is slower still by an additional factor equal to the number of input units. Parallel perturbation allows faster learning than sequential perturbation, by a factor that does not depend on network size. We also characterize how uncertainty in quantities used in the stochastic updates affects the learning curves. This study suggests that in practice, weight perturbation may be slow for large networks, and node perturbation can have performance comparable to that of direct gradient descent when there are few output units. However, these statements depend on the specifics of the learning problem, such as the input distribution and the target function, and are not universally applicable. PMID- 16212769 TI - Information geometry of interspike intervals in spiking neurons. AB - An information geometrical method is developed for characterizing or classifying neurons in cortical areas, whose spike rates fluctuate in time. Under the assumption that the interspike intervals of a spike sequence of a neuron obey a gamma process with a time-variant spike rate and a fixed shape parameter, we formulate the problem of characterization as a semiparametric statistical estimation, where the spike rate is a nuisance parameter. We derive optimal criteria from the information geometrical viewpoint when certain assumptions are added to the formulation, and we show that some existing measures, such as the coefficient of variation and the local variation, are expressed as estimators of certain functions under the same assumptions. PMID- 16212771 TI - Hairpins in bookstacks: information retrieval from biomedical text. AB - Current advances in high-throughput biology are accompanied by a tremendous increase in the number of related publications. Much biomedical information is reported in the vast amount of literature. The ability to rapidly and effectively survey the literature is necessary for both the design and the interpretation of large-scale experiments, and for curation of structured biomedical knowledge in public databases. Given the millions of published documents, the field of information retrieval, which is concerned with the automatic identification of relevant documents from large text collections, has much to offer. This paper introduces the basics of information retrieval, discusses its applications in biomedicine, and presents traditional and non-traditional ways in which it can be used. PMID- 16212772 TI - Text mining and ontologies in biomedicine: making sense of raw text. AB - The volume of biomedical literature is increasing at such a rate that it is becoming difficult to locate, retrieve and manage the reported information without text mining, which aims to automatically distill information, extract facts, discover implicit links and generate hypotheses relevant to user needs. Ontologies, as conceptual models, provide the necessary framework for semantic representation of textual information. The principal link between text and an ontology is terminology, which maps terms to domain-specific concepts. This paper summarises different approaches in which ontologies have been used for text mining applications in biomedicine. PMID- 16212773 TI - Information retrieval and knowledge discovery utilising a biomedical Semantic Web. AB - Although various ontologies and knowledge sources have been developed in recent years to facilitate biomedical research, it is difficult to assimilate information from multiple knowledge sources. To enable researchers to easily gain understanding of a biomedical concept, a biomedical Semantic Web that seamlessly integrates knowledge from biomedical ontologies, publications and patents would be very helpful. In this paper, current research efforts in representing biomedical knowledge in Semantic Web languages are surveyed. Techniques are presented for information retrieval and knowledge discovery from the Semantic Web that extend traditional keyword search and database querying techniques. Finally, some of the challenges that have to be addressed to make the vision of a biomedical Semantic Web a reality are discussed. PMID- 16212774 TI - Extraction of biological interaction networks from scientific literature. AB - Biology can be regarded as a science of networks: interactions between various biological entities (eg genes, proteins, metabolites) on different levels (eg gene regulation, cell signalling) can be represented as graphs and, thus, analysis of such networks might shed new light on the function of biological systems. Such biological networks can be obtained from different sources. The extraction of networks from text is an important technique that requires the integration of several different computational disciplines. This paper summarises the most important steps in network extraction and reviews common approaches and solutions for the extraction of biological networks from scientific literature. PMID- 16212775 TI - Online tools to support literature-based discovery in the life sciences. AB - In biomedical research, the amount of experimental data and published scientific information is overwhelming and ever increasing, which may inhibit rather than stimulate scientific progress. Not only are text-mining and information extraction tools needed to render the biomedical literature accessible but the results of these tools can also assist researchers in the formulation and evaluation of novel hypotheses. This requires an additional set of technological approaches that are defined here as literature-based discovery (LBD) tools. Recently, several LBD tools have been developed for this purpose and a few well motivated, specific and directly testable hypotheses have been published, some of which have even been validated experimentally. This paper presents an overview of recent LBD research and discusses methodology, results and online tools that are available to the scientific community. PMID- 16212776 TI - The next generation of literature analysis: integration of genomic analysis into text mining. AB - Text-mining systems are indispensable tools to reduce the increasing flux of information in scientific literature to topics pertinent to a particular interest in focus. Most of the scientific literature is published as unstructured free text, complicating the development of data processing tools, which rely on structured information. To overcome the problems of free text analysis, structured, hand-curated information derived from literature is integrated in text-mining systems to improve precision and recall. In this paper several text mining approaches are reviewed and the next step in development of text-mining systems, which is based on a concept of multiple lines of evidence, is described: results from literature analysis are combined with evidence from experiments and genome analysis to improve the accuracy of results and to generate additional knowledge beyond what is known solely from literature. PMID- 16212777 TI - Get ready to GO! A biologist's guide to the Gene Ontology. AB - The Gene Ontology (GO) project provides a controlled vocabulary to facilitate high-quality functional gene annotation for all species. Genes in biological databases are linked to GO terms, allowing biologists to ask questions about gene function in a manner independent of species. This tutorial provides an introduction for biologists to the GO resources and covers three of the most common methods of querying GO: by individual gene, by gene function and by using a list of genes. [For the sake of brevity, the term 'gene' is used throughout this paper to refer to genes and their products (proteins and RNAs). GO annotations are always based on the characteristics of gene products, even though it may be the gene that is cited in the annotation.]. PMID- 16212778 TI - Phenolic acids analysis in ligusticum chuanxiong using HPLC. AB - A reversed-phase high-performance liquid chromatographic method with diode array UV detection is developed for the determination of five kinds of phenolic acids common in herbal medicines. Based on this method, ferulic acid and caffeic acid are found to be two main phenolic acids in Chuanxiong (one of the important crude drugs in traditional Chinese medicine). More important, ferulic acid is found to exist in free form, and caffeic acid--a previously unreported component--is found to exist in esterified or insoluble-bound form. PMID- 16212779 TI - Modified Diatomaceous earth as a principal stationary phase component in TLC. AB - Modified natural diatomaceous earth (DE) is a principal component of the stationary phase in normal thin-layer chromatography (TLC) applications and is mixed with commercial silica gel 60GF254 (Si-60GF254). Modification is carried out by flux calcination and refluxing with acid. Natural DE, modified DEs [flux calcinated (FC)DE and FCDE-I), and Si-60GF254 are characterized by scanning electron microscopy and Fourier-transform-IR spectroscopy. Particle size, specific surface area, pore distribution, pore volume, and surface hydroxyl group density parameters of materials are determined by various techniques. FCDE-I and Si-60GF254 are investigated for their usefulness in the stationary phase of TLC both individually and in composition. Commercially available red and blue ink samples are run on layers of Si-60GF254 and FCDE-I individually, and on various FCDE-I and Si-60GF254 mixtures. Butanol-ethanol-2M ammonia (3:1:1, v/v) and butanol-acetic acid-water (12:3:5, v/v) mixtures are used as mobile phases. The polarities of stationary phases decrease, and the retention factor (Rf) values of ink components increase when the FCDE-I content of the stationary phase increases. The properties of the stationary phase can be optimized by adding FCDE I to Si-60GF254. This study may be useful in understanding both the systematic effects of stationary phase properties [e.g., specific surface area and surface hydroxyl group density, aOH(s)] and those of the mobile phase (e.g., polarity and acidity) on Rf values and the separability of components. PMID- 16212780 TI - Identification of unknown quaternary ammonium compounds in corneal epithelium and aqueous humor. AB - Bovine corneal epithelium and bovine aqueous humor are investigated for their content of quaternary ammonium compounds. In total, four compounds are found. Three of these are identified. For the fourth compound, a proposal for its structure is made on the basis of tandem mass spectrometry fragmentation spectra. The compounds investigated have m/z values of 146, 160, and 174. The compounds with m/z 146 are confirmed as acetylcholine (in corneal epithelium) and (3 carboxypropyl)-trimethylammonium (in both corneal epithelium and aqueous humor). The compound with m/z 174 is identified as butyrylcholine (in corneal epithelium). The compound with m/z 160 is probably acetyl-g-homocholine (in both corneal epithelium and aqueous humor). For both butyrylcholine and acetyl-g homocholine, it is the first time the presence of these compounds in corneal epithelium or aqueous humor (or both) is described. Both acetylcholine and butyrylcholine are unstable compounds, which are probably susceptible to enzymatic degradation by acetylcholine-esterase and butytrylcholine- esterase, respectively. PMID- 16212781 TI - The ICH guidance in practice: stress degradation studies on stavudine and development of a validated specific stability-indicating HPTLC assay method. AB - A sensitive, selective, precise, and stability-indicating high-performance thin layer chromatographic (HPTLC) method for the analysis of stavudine both as a bulk drug and in formulations is developed and validated. The solvent system consisted of toluene-methanol-chloroform-acetone (7.0:3.0:1.0:1.0, v/v/v/v). Densitometric analysis of stavudine is carried out in the absorbance mode at 270 nm. This system is found to give compact spots for stavudine (retention factor value of 0.45 +/- 0.05) following development of chromatoplates with the mobile phase. Stavudine is subjected to acid and alkali hydrolysis, oxidation, dry-heat and wet heat treatment, and photo and UV degradation. The drug undergoes degradation under acidic and basic conditions, oxidation, and wet-heat degradation. Linearity is found to be in the range of 30-1000 ng/spot with a significantly high value of correlation coefficient. The linear regression analysis data for the calibration plots show a good linear relationship with r2 = 0.9997 +/- 0.05 in the working concentration range of 300 to 1000 ng/spot. The mean value of slope and intercept are 0.10 +/- 0.06 and 22.12 +/- 1.08, respectively. The method is validated for precision, robustness, and recovery. The limits of detection and quantitation are 10 and 30 ng/spot, respectively. The proposed HPTLC method is utilized to investigate the kinetics of the acid degradation process. Arrhenius plot is constructed and activation energy is calculated. PMID- 16212782 TI - RP-HPLC method for the quantitative analysis of naturally occurring flavonoids in leaves of Blumea balsamifera DC. AB - A selective and sensitive reversed-phase (RP) high-performance liquid chromatographic method is developed for the quantitative analysis of five naturally occurring flavonoids of Blumea balsamifera DC, namely dihydroquercetin 7,4'-dimethyl ether (DQDE), blumeatin (BL), quercetin (QN), 5,7,3',5' tetrahydroxyflavanone (THFE), and dihydroquercetin-4'-methyl ether (DQME). These compounds have been isolated using various chromatographic methods. The five compounds are completely separated within 35 min using an RP C18, Nucleosil column and with an isocratic methanol-0.5% phosphoric acid (50:50, v/v) mobile phase at the flow rate of 0.9 mL/min. The separation of the compounds is monitored at 285 nm using UV detection. Identifications of specific flavonoids are made by comparing their retention times with those of the standards. Reproducibility of the method is good, with coefficients of variation of 1.48% for DQME, 2.25% for THFE, 2.31% for QN, 2.23% for DQDE, and 1.51% for BL. The average recoveries of pure flavonoids upon addition to lyophilized powder and subsequent extraction are 99.8% for DQME, 99.9% for THFE, 100.0% for BL, 100.6% for DQDE, and 97.4% for QN. PMID- 16212783 TI - Determination of four active saponins of Panax notoginseng in rat feces by high performance liquid chromatography. AB - A method is developed for the determination of ginsenoside Rg1, Rb1, Rd, and notoginsenoside R1 of Panax notoginseng (PNS) in rat feces after oral and intravenous administration of total saponins of PNS. The fecal samples are treated with organic extraction and solid-phase extraction prior to high performance liquid chromatography. The calibration curves for the four saponins are linear in the given concentration ranges. The precision of the method is in the range of 1.0-10.0% (relative standard deviation), and the accuracy is between 80.0% and 110%. The recoveries of this method are all over 75%. This method is successfully applied to the analyses of fecal samples of rats treated with PNS. PMID- 16212784 TI - HPLC determination and pharmacokinetics of osthole in rat plasma after oral administration of Fructus Cnidii extract. AB - A simple and sensitive high-performance liquid chromatographic (HPLC) method is developed for the determination of osthole in rat plasma and applied to a pharmacokinetic study in rats after administration of Fructus Cnidii extract. After addition of fluocinonide as an internal standard, plasma samples are extracted with diethyl ether. HPLC analysis of the extracts is performed on a Hypersil ODS2 analytical column, using methanol-0.4% acetic acid (65:35, v/v) as the mobile phase. The UV detector is set at 322 nm. The standard curve is linear over the range 0.0520-5.20 microg/mL (r = 0.9979). The mean extraction recoveries of osthole at three concentrations were 81.0%, 91.2%, and 90.7%, respectively. The intra- and interday precisions have relative standard deviations from 1.9% to 4.9%. The limit of quantitation is 0.0520 microg/mL. The HPLC method developed can easily be applied to the determination and pharmacokinetic study of osthole in rat plasma after the animals are given the Fructus Cnidii extract. The plasma concentration of osthole from six rats showed a Cmax of 0.776 +/- 0.069 microg/mL at Tmax of 1.0 +/- 0.3 h. PMID- 16212785 TI - Determination of meropenem by capillary electrophoresis using direct injection of serum. AB - Concentration determination of meropenem, a carbapenem antibiotic, using a capillary electrophoresis method by direct injection of serum samples without any pretreatment is described herein. Sodium tetraborate (25 mM)-sodium hydroxide (0.1 M) containing sodium dodecyl sulfate (90 mM) is used as a run buffer (pH 10.0). Meropenem is detected at its absorption maximum at 297 nm. Migration time of meropenem is approximately 7.2 min, and the detection limit of the assay is 2.0 mg/L at a signal-to-noise ratio of 3.0. The relative standard deviations of intra- and interassay accuracies are 3.43-8.87% and 4.28-8.54%, respectively, at a nominal concentration of 6.3-100.0 mg/L, and their recovery rates are 94-111% and 92-105%, respectively. PMID- 16212786 TI - Validation of an HPLC method for quantitation of MDMA in tablets. AB - An isocratic reversed-phase high-performance liquid chromatography (HPLC) method is developed and validated for the quantitation of 3,4 methylenedioxymethamphetamine (MDMA) in tablets. The chromatographic separation is achieved with potassium phosphate buffer (pH 3.2)-acetonitrile (9:1, v/v) as mobile phase, a Chromspher B column, and UV detection at 210 nm. The calibration curve is linear from 1.4 to 111 microg/mL. The percent relative standard deviation for intra- and interday precision studies is 2.7% each. The measurement uncertainty is estimated to 9%. The method is specific and successfully used for routine quantitation of MDMA in tablets. PMID- 16212787 TI - Urine steroid profile of judo competitors affected by acute physical exercises. AB - A heterogeneous group of 10 male and 15 female judo players are utilized in this study. The subjects complete a standardized maximal treadmill exercise test. Urine samples are collected at the pre- and postexercise stages. The urine steroids are measured using a gas chromatography-mass spectrometry instrument. In rest and after exercise, significantly higher testosterone and epitestosterone concentrations in males (p < 0.01) are found. The etiocholanolone dehydroepiandrosterone (DHEA) ratio is significantly lower in males than females (p < 0.05). In both males and females, etiocholanolone concentration significantly decreases with the effect of exercise (p < 0.05). 11-OH etiocholanolone concentration also significantly decreases, but only in females (p < 0.05). Positive correlation is found between the changes of the etiocholanolone and epitestosterone concentration caused by exercise. PMID- 16212788 TI - A validated HPLC method for the quantitative analysis of trans-resveratrol and trans-piceid in Hungarian wines. AB - A new, simple procedure for the determination of phenolic components of wines, such as resveratrol and piceid isomers, has been elaborated and validated. A set of 70 red wines and 3 white wines from two wineries (Polgar Winery and Bock Winery, Villany, Hungary) are analyzed by reversed-phase high-performance liquid chromatography. The samples are injected without pretreatment and UV-vis and mass spectrometric (MS) detection has been applied. The detection limit for trans resveratrol and for trans-piceid is found to be 0.9 and 0.6 pmol for the UV-vis detection method and 0.3 and 0.2 pmol for the MS detection method. Trans resveratrol and trans-piceid are found in red wines from 0.1 to 14.3 mg/L and from 3.8 to 16.4 mg/L concentrations, respectively. PMID- 16212790 TI - HPLC characterization of betalains from plants in the amaranthaceae. AB - HPLC characterization of reversed-phase (RP) high-performance liquid chromatography (HPLC) has been widely used in separation and identification of plant pigments. An effective RP-HPLC-based method is established to systematically isolate, identify, and quantitate the betalain pigments in the plants of 37 species of eight genera in the Amaranthaceae. A total of 16 betacyanins and three betaxanthins are characterized mainly using the RP-HPLC method and also with the aid of mass spectroscopy. The identified betacyanins include eight amaranthine-types, six gomphrenin-types, and two betanin-types. They are also divided into six simple (nonacylated) betacyanins and 10 acylated betacyanins. Acylated betacyanins are identified as betanidin 5-O-beta glucuronosylglucoside or betanidin 6-O-beta-glucoside acylated with ferulic, p coumaric, or 3-hydroxy-3-methylglutaric acids. Three betaxanthins were separated from Celosia species in the Amaranthaceae and identified to be immonium conjugates of betalamic acid with dopamine, 3-methoxytyramine, and (S) tryptophan; the latter two are found to be new betaxanthins from plants. PMID- 16212789 TI - Simultaneous determination of vincristine, vinblastine, catharanthine, and vindoline in leaves of catharanthus roseus by high-performance liquid chromatography. AB - A simple reversed-phase liquid chromatographic method is developed for the simultaneous quantitation of the anticancerous drugs vincristine, vinblastine, and their precursors catharanthine and vindoline using a Merck Chromolith Performance reversed-phase high-performance liquid chromatography column. A better resolution is obtained in comparison with available particulate-type C18 columns. The column provides good reproducibility and peak symmetry. Chromatography is carried isocratically with a mobile phase of acetonitrile-0.1M phosphate buffer containing 0.5% glacial acetic acid (21:79, v/v; pH 3.5) at a flow rate of 1.2 mL/min and UV detection at 254 nm. Parameters such as linearity, limits of quantitation (LOQ) and detection (LOD), precision, accuracy, recovery, and robustness are studied. The method is selective and linear for alkaloid concentration in the range 0.25 microg-25 microg/mL. The LOQ and LOD are 25, 46, 56, and 32 microg/mL and 8, 14, 18, and 10 microg/mL, respectively. The results of accuracy studies are good. Values for coefficient of variation are 2.50, 1.82, 1.33, and 1.13, respectively. The percent recovery of the alkaloids was found to be 96%, 97%, 98%, and 98%, respectively. Peak purity and homogeneity of these compounds in plant extract is studied using a photodiode-array detector. This simple and rapid method of analysis is applied for the determination of these alkaloids in a large number of leaf extracts of Catharanthus roseus.. PMID- 16212791 TI - Simultaneous determination of Sudan dyes and carotenoids in red pepper and tomato products by HPLC. AB - To simultaneously detect Sudan dyes and carotenoids in red pepper and tomato products, high-performance liquid chromatography (HPLC) methods with photodiode array detection are developed and validated. The methods include the use of end capped and nonend-capped adsorbents with a gradient elution system starting with water containing methanol. Water content of 9% in the starting mobile phase is found to be necessary to ensure sufficient separation of Sudan dyes and to avoid overlapping or interference with the carotenoids of considerable content. The data of the validation reveal the accuracy and precision of the developed methods. A limit of Sudan dyes detection of 1-5 microg/g in red pepper or tomato sauce could be approached. The methods provide excellent separation of the carotenoids from the unsaponified extracts of red pepper and the tomato products. PMID- 16212793 TI - Chromatographic determination of the effect of storage on lycopene. AB - A reverse-phase high-performance liquid chromatography (HPLC) method for the determination of lycopene in an alkaline lipid phase is described, and pigment stability in stored tomato sausage is reported. To avoid and replace the use of nitrite, lycopene from tomato products is added to minced meat and a tomato sausage with natural color is produced. Tomato sausage with and without nitrite were smoked in a smoking compartment and stored (4 degrees C and 8 degrees C) for 25 and 17 days, respectively. Among other factors, the quality of the tomato sausage depends on stability of lycopene during process and storage. Lycopene, being lipophilic, is extracted together with the polar and neutral fat in food. Efforts to purify lycopene from the fatty content will result in loss of pigment. The triacylglycerides obstruct the detection of lycopene by spectrophotometry or by HPLC with diode-array detection. To solve this problem, the triacylglycerides are hydrolyzed to free fatty acids just before analyzing lycopene on a column tolerating alkaline samples. At the end of the storage, loss of pigment in the sausage without nitrite was 26% stored at 4 degrees C and 19% at 8 degrees C. Corresponding results for the sausage with nitrite added as well as tomato paste show the loss of pigment is 20% and 45%. For each type of fatty food extracted, it is important to minimize the use of alkaline solutions because the HPLC equipment may be susceptible to alkaline conditions. PMID- 16212792 TI - Chromatographic determination of changes in pigments in spinach (Spinacia oleracea L.) during processing. AB - The content of individual chlorophyll and carotenoid pigments is determined in three spinach varieties (Lorelei, Springfield, and Ballet) after processing. Raw spinach and spinach that is steam-blanched for 3, 9, or 15 min is stored frozen at -24 degrees C for 6 months. In addition, spinach is air-dried at 75 degrees C, packed in atmospheric air or nitrogen, and stored at ambient temperature for 6 months. Processing has a significant effect on the content of individual chlorophyll and carotenoid pigments; however, there are no differences between varieties in their content of total and individual pigments in raw, frozen spinach. Increasing blanching time resulted in decreased contents of chlorophyll a and b and increased contents of chlorophyll a' and b' and pheophytin a and b because of pheophytinization. Changes in the color because of pheophytinization are only detected after 15 min blanching. The carotenoid pigments are more stable than the chlorophyll pigments during blanching. (all-E)-Violaxanthin is significantly reduced, caused by degradation to other xanthophylls, such as neochrome, during blanching. There are no significant differences in the content of chlorophyll a and b of dried spinach and blanched, frozen spinach. Formation of chlorophyll a' and b', pheophytin a and b, and chlorophyll a-1 and b-1 is observed after drying. The content of pheophytin a and b is significantly lower in dried versus blanched frozen samples. In dried spinach that is stored in atmospheric air, the content of beta-carotene [599 mg/kg dry matter (DM)] is significantly lower compared with nitrogen (766 mg/kg DM), and the content of (all-E)-lutein is lower than in blanched frozen spinach. Neochrome is not detected in raw spinach but in steam-blanched and dried spinach. No differences are observed in the content of (all-E)-neoxanthin, (9'Z)-neoxanthin, (all-E) violaxanthin, (all-E)-lutein epoxide, or neolutein A and B between spinach that is stored frozen after 3 min blanching and dried spinach. PMID- 16212794 TI - HPLC determination of the composition and stability of blackcurrant anthocyanins. AB - High-performance liquid chromatography with UV and mass spectrometry detectors are used to monitor the composition and stability of anthocyanins in blackcurrants harvested from different Ribes nigrum breeds at various ripeness phases. The highest amounts of pigments are found in overripe berries. The concentration of anthocyanins is higher in the berries of late blackcurrant breeds (Vakariai and Ben Alder). Delphinidin- 3-rutinoside is the dominant component in the reddish color berries (onset of ripening), and cyanidin-3 rutinoside is a major pigment in the black ones (ripe berries). Studies of the effect of temperature and light on the stability of the main pigments in blackcurrants show that aqueous solution prepared from a dry colorant is more stable when compared with the liquid water and ethanol extracts of coloring substances. Cyanidin-3-rutinoside is found to be the most thermally stable anthocyanin. PMID- 16212795 TI - Characterization of anthocyanin extracts from maize kernels. AB - The aim of the present work is to characterize the pigments present in the kernel of four native maize varieties related to the races Arrocillo, Conico, Peruano, and Purepecha to determine their possible use as natural dyes. Total anthocyanin content is determined by a conventional spectrophotometric method, and anthocyanin analysis is done by high-performance liquid chromatography. The stability of the pigment at pH is also evaluated. The four maize samples contained anthocyanin in both the pericarp and aleurone layer. Total anthocyanin content among samples ranged from 54 mg/100 g of sample to 115 mg/100 g of sample. Anthocyanin profiles are almost the same among the four samples. Differences are observed only in the relative percentage of each anthocyanin. The anthocyanins identified are cyanidin-3-glucoside, pelargonidin- 3-glucoside, peonidin-3-glucoside, cyanidin-3-(6" malonylglucoside), and cyanidin-3-(3",6" dimalonylglucoside). Anthocyanin extracts showed similar behavior in solutions with different pH. From pH 1-6 lambdamax values are maintained almost constant; however, above this pH value, a marked increase is observed in the bathochromic shifts, but the bluish color did not continue to change above pH 8. PMID- 16212796 TI - Isolation of flavanol-anthocyanin adducts by countercurrent chromatography. AB - Pigments of the flavanol-anthocyanin (F-A+) type detected earlier in wine are synthesized using a protocol adapted from the synthesis of procyanidin dimers. The F-A+ adduct thus obtained is purified by countercurrent liquid-liquid partition, currently referred to as countercurrent chromatography (CCC). The solvent system consists of tert-butyl methyl ether-n-butanol-acetonitrile-water (2:2:1:5, acidified with 0.1% trifluoroacetyl) with the light organic phase acting as a stationary phase and the aqueous phase as the mobile phase. Four fractions are recovered and analyzed by high-performance liquid chromatography coupled to a diode-array detector and electrospray ionization mass spectrometer. The multilayer CCC method allowed the separation of pigments in three different groups. The first group consists of hydrosoluble pigments present in fraction 1; the second group consists of the F-A+ adducts [catechin-malvidin 3 glucoside (Mv3glc), along with some (catechin)2-Mv3glc]; and the third group is their anthocyanin precursor, Mv3glc. PMID- 16212797 TI - Analysis of conjugated linoleic acids as 9-anthrylmethyl esters by reversed-phase high-performance liquid chromatography with fluorescence detection. AB - A simple and highly sensitive method for determining the fatty acid composition of food lipids containing conjugated linoleic acid (CLA) is described. The method is based on the separation of the 9-anthrylmethyl ester derivatives of saturated and unsaturated (conjugated and non-conjugated) fatty acids by reversed-phase high-performance liquid chromatography with fluorescence detection. Just like the other fatty acids, CLA reacts readily with 9-anthryldiazomethane at room temperature to produce 9-anthrylmethyl esters without isomerization and decomposition of the conjugated double bonds. Clear resolution of the individual fatty acids as their 9-anthrylmethyl esters is achieved on a highly efficient octadecylsilylated silica column (150- x 3-mm i.d., 3-microm particle size) using a stepwise gradient elution with methanol-water. The method is standardized with commercially available CLA isomers (cis-9, trans-11 and trans-10, cis-12 octadecadienoic acids, and their cis,cis and trans,trans isomers) and applied for determination of the fatty acid compositions of milk and sdairy products. PMID- 16212798 TI - High-level cerebellar expression of cytokines and adhesion molecules in fatal, paediatric, cerebral malaria. AB - Although the roles played by systemic tumour necrosis factor (TNF) and interleukin 1beta (IL-1beta), and their upregulation of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and E-selectin, in the pathogenesis of human cerebral malaria (CM) are well established, the role of local cytokine release, in the brain, remains unclear. Immunohistochemistry was therefore used to compare the expression of ICAM-1, VCAM-1, E-selectin, IL 1beta, TNF and transforming growth factor beta (TGF-beta) at light-microscope level, in cryostat sections of cerebral, cerebellar and brainstem tissues collected, post-mortem, from Ghanaian children. Among the 21 children investigated were 10 cases of CM, five of severe malarial anemia (SMA), one of purulent bacterial meningitis (PBM), two of non-central-nervous-system infection (NCNSI) and three children who had no infection (NI) when they died. Parasitised erythrocytes were detected in all of the sections from the cases of fatal malaria (CM and SMA), and sequestered leucocytes were present in most of the sections from the CM cases (but none of the sections from the SMA cases). Significantly elevated vascular expression of all three adhesion molecules investigated was detected in the brains of the 15 cases of fatal malaria and one of the cases of NCNSI (a child with Salmonella septicaemia), and in the malaria cases this showed highly significant co-localization with the areas of erythrocyte sequestration. In terms of the levels of expression of ICAM-1, VCAM-1 and E-selectin, there were, however, negligible differences between the CM and SMA cases. Although TGF beta showed intravascular and perivascular distribution in all the subjects, its expression was most intense in the PBM case and the CM group. Only in the sections from the PBM and CM cases did TNF and IL-1beta show prominent brain parenchymal staining, in addition to the intravascular and perivascular staining seen in all subjects. The highest observed expression of each of the six antigens studied was in the cerebellar sections of the malaria cases. Endothelial activation in the brain therefore appears to be a feature of fatal malaria and Salmonella sepsis, and in cases of fatal malaria is closely associated with leucocyte sequestration. In the present study, IL-1beta and TNF were only up regulated in the brains of children with neurodegenerative lesions, whereas TGF beta was present in all cases. PMID- 16212799 TI - The types and timing of the degenerative changes seen in the cysts during and after benzimidazole treatment of cystic echinococcosis. AB - In a large-scale Bulgarian study, 122 patients with abdominal and/or lung echinococcosis were randomly selected for treatment with albendazole or mebendazole. The main aims were to evaluate the effect of each drug on the hydatid cysts, to follow the changes in cyst morphology during and after treatment, and to determine how quickly each drug produced the first, detectable, degenerative changes in the cysts. Follow-up was based on periodic ultrasonography, chest radiography and computed tomography. The abdominal cysts were categorised as small (<5 cm in diameter) or large. As albendazole treatment had effects that were almost identical to those of mebendazole treatment, the results for the two drugs were combined. In the lungs and, particularly, in the abdomen, the size of the cysts being treated influenced the character and timing of the degenerative changes seen in them. The initial change seen in each abdominal cyst was detachment of the endocyst, which occurred 1-3 months (small cysts) or 2-5 months (large cysts) after the initiation of treatment (P<0.05). The abdominal cysts then developed a hyper-echoic/hyper-dense appearance, became smaller, and finally disappeared 3.3-9.3 months (small cysts) or 5.6-13.9 months (large cysts) after treatment began (P<0.05). The first degenerative change noted in the lung cysts was cyst rupture, which occurred as early as day 10 of therapy but was generally observed 1 or 2 months after treatment began. After their complete evacuation, the ruptured lung cysts shrank and became deformed, some disappearing within 5-9 months of the initiation of treatment. The degenerative changes recorded, which began significantly earlier in the lung cysts than in abdominal cysts, indicate serious damage to the cysts and the parasiticidal, curative effect of each of the two benzimidazoles employed. PMID- 16212800 TI - Tick modulation of the in-vitro expression of adhesion molecules by skin-derived endothelial cells. AB - As a tick feeds, its saliva induces innate and acquired immune responses in the host, including leucocyte infiltration into the bite site. Tick salivary glands produce molecules, however, that counteract many host defences against blood feeding. The effects of salivary-gland extracts (SGE) of Dermacentor andersoni and Ixodes scapularis on the expression of various adhesion molecules [E selectin, P-selectin, intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1)] by the sEND.1 cell line (which is based on cells from the subcutaneous tissue of mice) have now been investigated in vitro. The effects were found to differ with the tick species. The SGE of D. andersoni significantly down-regulated the expression of ICAM-1, whereas a similar extract prepared from I. scapularis significantly reduced the expression of P-selectin and VCAM-1. Tick salivary proteins therefore appear to have direct effects on adhesion-molecule expression, in addition to their previously established roles in down-regulating the pro-inflammatory cytokines that activate endothelial cells. It remains unclear exactly how the reduction of adhesion-molecule expression in the host's endothelial cells benefits the feeding tick but it may alter leucocyte migration to the bite site and/or reduce antigen presentation by the endothelial cells. It may also modulate the interactions between the host's leucocytes and any tick-borne pathogens, during initial infection of the endothelium. PMID- 16212801 TI - Lipase and antibacterial activities of a recombinant protein from the accessory glands of female Phlebotomus papatasi (Diptera: Psychodidae). AB - The reproductive accessory glands of the adult female have a functional versatility in insects, contributing to reproduction in various ways. The major protein secreted by the accessory glands of female Phlebotomus papatasi (Diptera, Psychodidae) has already been characterised and named PhpaLIP (for P. papatasi lipase) because, in terms of its amino-acid sequence, it is very similar to a wide range of vertebrate lipases. The gene coding for PhpaLIP has now been cloned into a pQE30 vector and expressed in Escherichia coli. When the recombinant PhpaLIP was tested in vitro, it was found to have not only lipase-like activity (when p-nitrophenyl caprylate was used as the substrate) but also specific antibacterial activity against some Gram-positive and Gram-negative bacteria. The possible physiological roles of PhpaLIP in P. papatasi are discussed, in the light of these results. PMID- 16212802 TI - Phlebotomine sandflies (Diptera: Psychodidae) of the Atlantic forest in Recife, Pernambuco state, Brazil: the species coming to human bait, and their seasonal and monthly variations over a 2-year period. AB - In a study of the phlebotomine sandflies (Diptera: Psychodidae) in a forest reserve in Recife, Pernambuco state, north-eastern Brazil, the sandflies landing on human bait between 1.00 and 1.42 h after sunset were collected weekly for 2 years. Although 10,287 sandflies of 10 Lutzomyia species were collected, almost all (96.5%) of the sandflies caught were Lu. umbratilis. This species and several others caught are potential vectors of some of the Leishmania parasites that cause human disease. The recorded landing rate for Lu. umbratilis peaked, at the high level of 333.3 flies/person-hour, during the collections made in May 2003.The relative rarity in the collections of males of some of the species caught probably indicates that these species do not lek on their bloodmeal sources. It is likely that the sizes of the local populations of species that are not very anthrophilic, such as Lu. flaviscutellata, are much larger than indicated by the collections made on human bait. PMID- 16212803 TI - Laboratory and field tests of the effectiveness of the lemon-eucalyptus extract, Citridiol, as a repellent against land leeches of the genus Haemadipsa (Haemadipsidae). AB - Citridiol is an extract of the leaves of Corymbia citriodora (Myrtaceae), the lemon eucalyptus, and mostly consists of p-menthane-3,8-diol isomers. The effectiveness of this extract as a repellent against land leeches of the genus Haemadipsa (Haemadipsidae), primarily H. sylvestris, was tested in the laboratory and field, in Peninsular Malaysia. The formulation tested, Mosi-guard Natural spray, contained 40% (w/w) Citridiol in a base of ethanol, water and isopropanol. In the laboratory test, specimens of H. sylvestris that were placed within moist, untreated arenas enclosed by treated paper rings made numerous attempts to cross the rings but were prevented or delayed from crossing over, in a dose-dependent manner. Mortality was high among the leeches that attempted to cross over the paper rings that had been sprayed to saturation point but low among the leeches that attempted to cross over paper rings that had only been partially treated, with a droplet-spray. The field study was carried out using indices that were formulated to reflect the severity of leech attack and the degree of repellency. Heavy or moderate spraying of footwear and trouser legs (tucked into socks) not only gave complete protection against bites by H. sylvestris and H. picta but also provided high enough repellency to keep the treated footwear virtually free of leeches. Even a light spray greatly reduced the numbers of leeches on footwear and delayed their progression toward biting the test subjects, although it failed to prevent bites completely. There was no decline in the repellency of the Citridiol when hourly assessments were made over a 6-h test period in the field. The results of the study show that Citridiol is highly repellent as well as toxic to leeches, and can be effectively used to prevent leech bites in the field. PMID- 16212804 TI - Imported malaria in pregnancy: a retrospective study of 18 cases in Marseilles, France. PMID- 16212805 TI - A 'pseudo outbreak' of the contamination of blood cultures with Volvox globator. PMID- 16212807 TI - GNOSIS: guidelines for neuro-oncology: standards for investigational studies reporting of phase 1 and phase 2 clinical trials. AB - We present guidelines to standardize the reporting of phase 1 and phase 2 neuro oncology trials. The guidelines are also intended to assist with accurate interpretation of results from these trials, to facilitate the peer-review process, and to expedite the publication of important and accurate manuscripts. Our guidelines are summarized in a checklist format that can be used as a framework from which to construct a phase 1 or 2 clinical trial. PMID- 16212809 TI - Scatter factor/hepatocyte growth factor in brain tumor growth and angiogenesis. AB - The multifunctional growth factor scatter factor/hepatocyte growth factor (SF/HGF) and its receptor tyrosine kinase c-Met have emerged as key determinants of brain tumor growth and angiogenesis. SF/HGF and c-Met are expressed in brain tumors, the expression levels frequently correlating with tumor grade, tumor blood vessel density, and poor prognosis. Overexpression of SF/HGF and/or c-Met in brain tumor cells enhances their tumorigenicity, tumor growth, and tumor associated angiogenesis. Conversely, inhibition of SF/HGF and c-Met in experimental tumor xenografts leads to inhibition of tumor growth and tumor angiogenesis. SF/HGF is expressed and secreted mainly by tumor cells and acts on c-Met receptors that are expressed in tumor cells and vascular endothelial cells. Activation of c-Met leads to induction of proliferation, migration, and invasion and to inhibition of apoptosis in tumor cells as well as in tumor vascular endothelial cells. Activation of tumor endothelial c-Met also induces extracellular matrix degradation, tubule formation, and angiogenesis in vivo. SF/HGF induces brain tumor angiogenesis directly through only partly known mechanisms and indirectly by regulating other angiogenic pathways such as VEGF. Different approaches to inhibiting SF/HGF and c-Met have been recently developed. These include receptor antagonism with SF/HGF fragments such as NK4, SF/HGF, and c-Met expression inhibition with U1snRNA/ribozymes; competitive ligand binding with soluble Met receptors; neutralizing antibodies to SF/HGF; and small molecular tyrosine kinase inhibitors. Use of these inhibitors in experimental tumor models leads to inhibition of tumor growth and angiogenesis. In this review, we summarize current knowledge of how the SF/HGF:c-Met pathway contributes to brain tumor malignancy with a focus on glioma angiogenesis. PMID- 16212811 TI - Expression of hypoxia-inducible carbonic anhydrases in brain tumors. AB - Malignant brain tumors exhibit distinct metabolic characteristics. Despite high levels of lactate, the intracellular pH of brain tumors is more alkaline than normal brain. Additionally, with increasing malignancy, brain tumors display intratumoral hypoxia. Carbonic anhydrase (CA) IX and XII are transmembrane isoenzymes that are induced by tissue hypoxia. They participate in regulation of pH homeostasis by catalyzing the reversible hydration of carbon dioxide. The aim of our study was to investigate whether brain tumors of different histology and grade of malignancy express elevated levels of CA IX and XII as compared to normal brain. We analyzed 120 tissue specimens from brain tumors (primary and metastatic) and normal brain for CA IX and XII expression by immunohistochemistry, Western blot, and in situ hybridization. Whereas normal brain tissue showed minimal levels of CA IX and XII expression, expression in tumors was found to be upregulated with increased level of malignancy. Hemangioblastomas, from patients with von Hippel-Lindau disease, also displayed high levels of CA IX and XII expression. Comparison of CA IX and XII staining with HIF-1alpha staining revealed a similar microanatomical distribution, indicating hypoxia as a major, but not the only, induction factor. The extent of CA IX and XII staining correlated with cell proliferation, as indicated by Ki67 labeling. The results demonstrate that CA IX and XII are upregulated in intrinsic and metastatic brain tumors as compared to normal brain tissue. This may contribute to the management of tumor-specific acid load and provide a therapeutic target. PMID- 16212810 TI - The role of pericytes in blood-vessel formation and maintenance. AB - Blood vessels are composed of two interacting cell types. Endothelial cells form the inner lining of the vessel wall, and perivascular cells--referred to as pericytes, vascular smooth muscle cells or mural cells--envelop the surface of the vascular tube. Over the last decades, studies of blood vessels have concentrated mainly on the endothelial cell component, especially when the first angiogenic factors were discovered, while the interest in pericytes has lagged behind. Pericytes are, however, functionally significant; when vessels lose pericytes, they become hemorrhagic and hyperdilated, which leads to conditions such as edema, diabetic retinopathy, and even embryonic lethality. Recently, pericytes have gained new attention as functional and critical contributors to tumor angiogenesis and therefore as potential new targets for antiangiogenic therapies. Pericytes are complex. Their ontogeny is not completely understood, and they perform various functions throughout the body. This review article describes the current knowledge about the nature of pericytes and their functions during vessel growth, vessel maintenance, and pathological angiogenesis. PMID- 16212812 TI - Neural stem cell migration toward gliomas in vitro. AB - Various in vivo studies demonstrated a migration tendency of neural stem cells (NSCs) toward gliomas, making these cells a potential carrier for delivery of therapeutic genes to disseminated glioma cells. We analyzed which factors determine NSC migration and invasion in vitro. Conditioned media prepared from 10 different human glioma cell lines, as well as 13 different tumor-associated growth factors, were analyzed for their chemotactic effects on murine C17.2 NSCs. The growth factor receptor status was analyzed by reverse transcriptase polymerase chain reaction. Invasion of NSCs into multicellular tumor spheroids generated from 10 glioma cell lines was quantified. NSCs displayed a heterogeneous migration pattern toward glioma spheroids as well as toward glioma cell-conditioned medium. Chemotactic migration was stimulated up to fivefold by conditioned medium as compared to controls. In coculture assays, NSC invasion varied from single cell invasion into glioma spheroids to complete dissemination of NSCs into glioma spheroids of different cell lines. Among 13 different growth factors, scatter factor/hepatocyte growth factor (SF/HGF) was the most powerful chemoattractant for NSCs, inducing a 2.5-fold migration stimulation. An antibody against SF/HGF inhibited migratory stimulation induced by conditioned media. NSC migration can be stimulated by various growth factors, similar to glioma cell migration. The extent to which NSCs infiltrate three-dimensional glioma cell aggregates appears to depend on additional factors, which are likely to include cell-to-cell contacts and interaction with extracellular matrix proteins. PMID- 16212813 TI - Modeling prognosis for patients with malignant astrocytic gliomas: quantifying the expression of multiple genetic markers and clinical variables. AB - The disparate lengths of survival among patients with malignant astrocytic gliomas (anaplastic astrocytomas [AAs] and glioblastoma multiforme [GBM]) cannot be adequately accounted for by clinical variables (patient age, histology, and recurrent status). Using real-time quantitative reverse transcription-polymerase chain reaction, we quantified the expression of four genes that were putative prognostic markers (CDK4, IGFBP2, MMP2, and RPS9) in a set of 43 AAs, 41 GBMs, and seven adjacent normal brain tissues. We previously explicated the expression and prognostic value of PAX6, PTEN, VEGF, and EGFR in these glioma tissues and established a comprehensive prognostic model (Zhou et al., 2003). This study attempts to improve that model by including four additional genetic markers, which exhibited a differential expression (P < 0.001) among tumor grades and between tumor and normal tissues. By including eight log-scaled gene expression variables, three clinical variables, and interaction terms among the eight genes, we established a prognostic model that accounted for two thirds of the variation (R2) in survival for this set of patients. To improve the R2 of the model without compromising its clinical utility, our data demonstrated that incorporating genes from different pathways markedly strengthens the model. Spearman rank correlation analysis of gene expression demonstrated a statistically significant positive correlation (P < 0.01) between the expression of IGFBP2-MMP2 and IGFBP2-VEGF in GBMs, but not in AAs. This finding suggests that the expression of IGFBP2 is associated with pathways activated specifically in GBMs that result in enhancing invasiveness and angiogenesis. PMID- 16212816 TI - Outreach teaching--coming soon to a practice near you? PMID- 16212814 TI - ERCC1 and ERCC2 polymorphisms and adult glioma. AB - ERCC2 and ERCC1 are important in DNA nucleotide excision repair and lie on chromosome 19q13.3 near a putative glioma suppressor region. We genotyped constitutive variants ERCC1 C8092A and ERCC2 K751Q and R156R in approximately 450 adults with glioma and 500 controls from two independent population-based series, uniformly reviewed patients' tumors to determine histopathologic category, and determined a variety of tumor markers among astrocytic tumors. Odds ratios (ORs) for glioblastoma for those carrying two ERCC1 A alleles versus none or one were 1.67 in series 1 and 1.64 in series 2, which yielded a combined OR of 1.67 (95% CI, 0.93-3.02; P = 0.09), adjusted for age, gender, ethnicity, and series. Odds ratios for the ERCC2 variants were not consistently elevated or reduced for the two series in all cases versus controls. However, among whites, for those with ERCC2 K751Q genotype QQ versus QK/KK, the OR for nonglioblastoma histologies versus controls was 1.82 (95% CI, 0.97-3.44; P = 0.06). Also, among whites, glioma patients were significantly more likely than controls to be homozygous for variants in both ERCC1 C8092A and ERCC2 K751Q (OR, 3.2; 95% CI, 1.1-9.3). Given the numbers of comparisons made, these findings could be due to chance. However, the results might warrant clarification in additional series in conjunction with the nearby putative glioma suppressor genes (GLTSCR1 and GLTSCR2). PMID- 16212815 TI - Combined immunoradiotherapy induces long-term remission of CNS relapse of peripheral, diffuse, large-cell lymphoma after allogeneic stem cell transplantation: case study. AB - Relapse of peripheral non-Hodgkin's lymphoma (NHL) in the central nervous system commonly has a poor prognosis. Graft-versus-leukemia effects (GvL) contribute substantially to eradication of hematological malignancies after allogeneic stem cell transplantation. Few data are available describing GvL activity within the brain. We report the case of a man allografted for peripheral NHL. On day +83 after transplantation a CNS relapse of the lymphoma occurred. The brain was irradiated with 44 Gy, anti-CD20 antibodies were given, and the immunosuppression was withdrawn. Subsequently, limited-stage, chronic graft-versus-host disease occurred. The lymphoma regressed completely, and the patient has been in continuous complete remission for 30 months. The favorable course suggests substantial contribution of immunomodulation to excellent outcome. PMID- 16212817 TI - Developing the first dental care pathway: the oral health assessment. AB - In this paper the authors define the concept of care pathways and explain what they are and how they have been applied in various areas of healthcare. The authors discuss the benefits of the care pathway approach and outline the first national dental NHS (clinical) care pathway and its link to National Institute for Clinical Excellence (NICE) guidance on dental recall intervals. The authors go on to explain that as a result of one of the key recommendations of Options for Change, the concept of care pathways will be extended to the primary dental services of the NHS. The first care pathway will be the Oral Health Assessment (OHA). The authors describe the development of, and rationale for, the OHA and provide details of its planned introduction. The authors conclude that the OHA should act as the initial interface between the primary care team and the dental patient. In so doing it should enable a change in the focus of care, from treatment-based to more prevention-led, personalised care appropriate to the specific needs of individual patients. PMID- 16212820 TI - Pharmaceutical prescribing for children. Part 1. Introduction and general principles of prescribing for children. AB - This paper is the first in a series which will review the current literature and evidence upon which primary care dentists should base their decisions to prescribe medication for children. As such it will offer guidance and suggest current best practice. The series has been edited by Dr Nikolaus Palmer. This first paper deals with general principles. A second paper on prescribing for children with regard to pain and analgesia also appears in this edition of Primary Dental Care. Future papers will address prescribing for children with relation to: fungal and viral infections, the management of paediatric patients with cardiac problems, the management of immunocompromised patients, sedatives and hypnotics, dental trauma, emergency drugs, fluorides and fluoride supplements. PMID- 16212824 TI - Description of a system designed to assist primary dental care clinicians in decision-making with regard to specialist periodontal referrals and report of two clinical audits using the system. AB - INTRODUCTION: Identifying which patients are at risk of significant periodontal breakdown and may need specialist care is a process fraught with problems. This paper summarises factors that should be considered by referring clinicians, describes a system to assist them, and presents the results of two sequential audits carried out in general dental practice using the system. AIMS, MATERIALS AND METHODS: The author aims to present the system and the results of two sequential audits, carried out in the same eight general dental practices by 23 general dental practitioners (GDPs) after training from a specialist in periodontics. The system is based on the use of the Basic Periodontal Examination (BPE), a constant pressure periodontal probe, and a computer screen that prompts the clinician who is performing periodontal screening. A series of risk factors, including smoking, presence of relevant systemic disease and age, is used to modulate the score. Patients are then classified into one of three complexity groups. Those in the Complexity 3 group are deemed to be potentially at risk of significant periodontal breakdown and in need of referral for specialist advice and/or care, in accordance with the published British Society of Periodontology guidelines. In each audit a random sample of the records of ten dentate patients over the age of 20 years, who attended for recall or new examinations by each of the 23 GDPs over a calendar month, were assessed to see whether or not BPE scores had been recorded and whether or not those classified as Complexity 3 (and in need of referral) had been correctly identified and referred. RESULTS: In both audits the total random sample of patient records was 225 (ten for 22 dentists and five for the remaining dentist who only worked for a short time during the months of the audit). In the first audit cycle 139 (62%) of the random sample of records included a BPE score and 11 (8%) of the patients concerned were identified as Complexity 3 and referred for specialist advice and/or treatment. All 11 accepted the referral and when examined by the specialist in periodontics were confirmed as Complexity 3. In the second audit cycle, which took place seven months later, 179 (80%) of the random sample of patient records included a BPE score and 14 patients were assessed by their GDPs as being Complexity 3. All 14 accepted a referral and were deemed to be Complexity 3 when they were subsequently examined by the specialist in periodontics. CONCLUSION: Although the full potential of the system was not investigated during the two audits, these preliminary results suggest that it may assist referring clinicians in identifying which patients to refer and encourage clinicians to perform periodontal screening. PMID- 16212821 TI - Pharmaceutical prescribing for children. Part 2. Analgesia and prescribing for children in pain. AB - This paper, the second in a series to be published in Primary Dental Care, addresses the issues of prescribing analgesics for children and the control of pain. It is crucial to remember that in infants and children, as in adults, pain, anxiety and fear are often associated. All concerned with the care of children have a duty to anticipate and avoid, as well as to relieve pain. PMID- 16212825 TI - Launch of Key Skills in Primary Dental Care. AB - On the morning of Tuesday, 21st June 2005 an audience of approximately 150 attended the launch of the Faculty of General Dental Practice (UK)'s Key Skills in Primary Dental Care e-learning programme. This package consists of seven modules each of which deals with a skill that is fundamental to the safe practice of clinical dentistry, not just in primary care, as the title suggests, but all areas including primary and secondary care. The package is available as a series of seven CD-ROMs or online at a protected website. The seven key skills are: infection control, legislation and good practice guidelines, clinical record keeping, team training, radiography, risk management and communication, medical emergencies. The launch consisted of a series of short presentations followed by a question and answer session. PMID- 16212827 TI - An investigation into Merseyside General Dental Practitioners' interest in primary care research, their views on research and their training needs. AB - OBJECTIVES: The aims of this investigation were to evaluate the interests of primary care dental practitioners within the Mersey Postgraduate Deanery in research and their views on research, their experience and research training needs. METHOD: A questionnaire was sent to all practitioners in the Mersey Deanery asking for views on dental research, whether they had been involved in any research projects or had any research training, and whether they would be interested in research training and being part of a primary dental care research network. RESULTS: A total of 192 practitioners from the 1120 in the Mersey Deanery expressed an interest in being involved in primary care research. Most believed that primary care research was very important in providing a stronger evidence-base in dentistry and improved quality of dental care. Over 50% of respondents were interested in collaborative research, provided that their income and time could be protected and it was part of the normal working day. Almost 25% had some research experience and a number had undergone research training, ranging from informal training to part of a degree. CONCLUSIONS: A number of GDPs in the Mersey Deanery are interested in primary care research. With appropriate training, support and recognition within the new Personal Dental Services (PDS) contracts, there is a golden opportunity for more primary care dentists to participate in research. This, in time, will add to the evidence base in dentistry and should improve patient care. PMID- 16212831 TI - Telomere length among the elderly and oldest-old. AB - Human chromosomes terminate in a number of repeats of the sequence TTAGGG. At birth, each chromosome end is equipped with approximately 15 kb of telomere sequence, but this sequence is shortened during each cell division. In cell cultures telomere shortening is associated with senescence, a phenomenon that has also been observed in normal adult tissues, indicating that telomere loss is associated with organismal ageing. Previous work has established that the rate of telomere loss in humans is age dependent, and recent work shows a sex-specific difference in telomere length and shortening in individuals over the age span of 20 to 75 years. Here, terminal restriction fragment lengths on DNA purified from whole blood were measured to examine the mean telomere length in a cross sectional cohort of 816 Danish individuals of age 73 to 101 years. In this age group, females show a linear correlation between telomere length and age, whereas the pattern tends to be nonlinear (quadratic in age) for males. This difference in telomere length dynamics between the 2 sexes may be caused by several different mechanisms, including differences in selection by mortality, differences in leukocyte population or different telomerase expression pattern. PMID- 16212832 TI - The heritability of telomere length among the elderly and oldest-old. AB - A tight link exists between telomere length and both population doublings of a cell culture and age of a given organism. The more population doublings of the cell culture or the higher the age of the organism, the shorter the telomeres. The proposed model for telomere shortening, called the end replication problem, explains why the telomere erodes at each cellular turnover. Telomere length is regulated by a number of associated proteins through a number of different signaling pathways. The determinants of telomere length were studied using whole blood samples from 287 twin pairs aged 73 to 95 years. Structural equation models revealed that a model including additive genetic effects and non-shared environment was the best fitting model and that telomere length was moderately heritable, with an estimate that was sensitive to the telomere length standardization procedure. Sex-specific analyses showed lower heritability in males, although not statistically significant, which is in line with our earlier finding of a sex difference in telomere dynamics among the elderly and oldest old. PMID- 16212833 TI - Subjective wellbeing and sleep problems: a bivariate twin study. AB - The relationship between subjective wellbeing (SWB) and self-reported sleep problems was investigated in a cohort of Norwegian twins aged 18 to 31 years. Questionnaire data from 8045 same- and opposite-sex twins were analyzed using structural equation modeling to explore the relative effects of genetic and environmental influences on phenotypic variance and covariance. Special attention was paid to sex-specific effects. The correlation between the phenotypes was estimated to be -.43. Univariate analyses indicated considerable genetic influences for both SWB and sleep problems, for male and female twins alike. The best fitting bivariate model specified additive genetic and individual environmental factors for both phenotypes, and nonadditive genetic effects for sleep problems, with no sex-specific effects. Genetic and environmental effects accounted for 60% and 40% of the phenotypic correlation, respectively. Additive genetic factors affecting the two phenotypes were correlated (-.85), suggesting that part of the genetic effects that positively influence SWB also protect against sleep problems. In conclusion, the results indicate considerable overlap in genetic etiology for SWB and sleep problems, for males and females alike. PMID- 16212834 TI - Twin studies on obsessive-compulsive disorder: a review. AB - Genetic factors have historically been thought of as important in the development of obsessive-compulsive disorder (OCD). For the estimation of the relative importance of genetic and environmental factors, twin studies are an obvious approach. Twin studies of OCD have a long history, starting in 1929. In this review, over 70 years of twin research of OCD is presented, using four different approaches that represent the steps in the twin research of OCD from past to present. These steps include (1) case-studies of twins with OCD from the old literature; (2) twin studies of OCD using Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria; (3) twin studies of OCD using a dimensional approach, comparing resemblances in monozygotic and dizygotic twins; and (4) twin studies of OCD using a dimensional approach, analyzing the data with Structural Equation Modeling. It is concluded that only the studies using the last method have convincingly shown that, in children, obsessive-compulsive (OC) symptoms are heritable, with genetic influences in the range of 45% to 65%. In adults, studies are suggestive for a genetic influence on OC symptoms, ranging from 27% to 47%, but a large twin study using a biometrical approach with continuous data is still needed to provide conclusive evidence. Strategies for future twin studies of OCD are discussed. PMID- 16212835 TI - Comorbidity between ADHD and symptoms of bipolar disorder in a community sample of children and adolescents. AB - The prevalence and frequency of comorbidity of possible bipolar disorder was examined with attention-deficit hyperactivity disorder (ADHD) in a nonreferred population of twins. Children and adolescents aged 7 to 18 years with a history of manic symptoms were identified from a population-based twin sample obtained from state birth records (n = 1610). The sample was enriched for ADHD; however, there was also a random control sample (n = 466), which allowed a look at the population prevalence of the disorder. Juveniles with threshold or below threshold manic episodes were further assessed for comorbidity with Diagnostic and Statistical Manual of Mental Disorders (4th ed.; DSM-IV; American Psychiatric Association, 1994) and population-defined ADHD subtypes (from latent class analysis) using Fisher's exact test. Nine juveniles who exhibited DSM-IV manic (n = 1), hypomanic (n = 2) or below threshold episodes (n = 6) were identified. The population prevalence of broadly defined mania in the random sample was 0.2%. The possible manic episodes showed significant comorbidity with population-defined severe combined and talkative ADHD subtypes. It can be concluded that there is a significant association of bipolar symptoms with two population-defined subtypes of ADHD. Episodes of possible bipolar disorders as defined by DSM-IV are uncommon in this nonreferred sample. Children and adolescents with ADHD appear to be only modestly at increased risk for bipolar disorders. PMID- 16212837 TI - Genetic contributions to subtypes of aggression. AB - Boys and girls may display different styles of aggression. The aim of this study was to identify subtypes of aggression within the Child Behavior Checklist (CBCL) aggression scale, and determine their characteristics for both sexes. Maternal CBCL ratings of 7449 7-year-old twin pairs were analyzed using principal components analyses to identify subtypes of aggression, and structural equation modeling to carry out genetic analyses. Two aggression subtypes were identified: relational and direct aggression. The correlation between these subtypes was .58 for boys and .47 for girls. Boys had higher mean scores for both subtypes of aggression, but sex differences were largest for direct aggression. For relational aggression, 66% of the variance was due to additive genetic influences, 16% to shared environment and 18% to nonshared environment. For direct aggression, additive genetic effects accounted for 53% of the variance in males and 60% in females, shared environment explained 23% of the variance in males and 13% in females, and nonshared environmental effects explained 24% of the variance in males and 27% in females. Covariance between the aggression subtypes was mostly accounted for by additive genetic (55% for boys, 58% for girls) and shared environmental influences (33% for boys, 30% for girls). Direct and relational aggression were both influenced by one underlying set of shared environmental factors, but only partly by the same genes (the genetic correlation was .54 for boys and .43 for girls). These findings may have implications for how aggressive behavior should be assessed in boys and girls. PMID- 16212836 TI - Eating disorders: from twin studies to candidate genes and beyond. AB - Substantial effort has been put into the exploration of the biological background of eating disorders, through family, twin and molecular genetic studies. Family studies have shown that anorexia (AN) and bulimia nervosa (BN) are strongly familial, and that familial etiologic factors appear to be shared by both disorders. Twin studies often focus on broader phenotypes or subthreshold eating disorders. These studies consistently yielded moderate to substantial heritabilities. In addition, there has been a proliferation of molecular genetic studies that focused on Diagnostic and Statistical Manual of Mental Disorders (4th ed.; DSM-IV; American Psychiatric Association, 1994) AN and BN. Seven linkage regions have been identified in genome-wide screens. Many genetic association studies have been performed, but no consistent association between a candidate gene and AN or BN has been reported. Larger genetic association studies and collaborations are needed to examine the involvement of several candidate genes and biological pathways in eating disorders. In addition, twin studies should be designed to assist the molecular work by further exploring genetic determinants of endophenotypes, evaluating the magnitude of contribution to liability of measured genotypes as well as environmental risk factors related to eating disorders. In this manner twin and molecular studies can move the field forward in a mutually informative way. PMID- 16212838 TI - Heritabilities of lipids in young European American and African American twins. AB - Twin studies of lipids have almost exclusively involved Caucasians. People of African descent are known to show a healthier lipid profile, but relatively little is known about ethnic differences in genetic and environmental influences on lipids. One hundred and six African American (AA) and 106 European American (EA) twins (30 singletons and 91 complete pairs: 49 monozygotic, 21 dizygotic and 21 opposite-sex) from the south-eastern United States were studied (mean age 17.9 +/- 3.2 years; 79% fasting). Lipids were assayed with the Cholestech LDX system. Analyses were adjusted for fasting status. Generalized estimating equations were used to test for the effects of sex and ethnicity on means, controlling for the dependence within twin pairs. Structural equation modeling was used to estimate genetic and environmental effects on each lipid variable. Females showed higher high-density lipoprotein (HDL) values than males (p < .001) and AAs showed higher HDL values than EAs (p < .001). EA males had higher triglyceride values than other groups (p = .02). All parameter estimates could be set equal across sex. Parameter estimates for total cholesterol, triglycerides and HDL could be set equal across ethnicity. The best fitting model for low-density lipoprotein (LDL) showed higher heritability in AAs (.92) than EAs (.69). Heritabilities ranged from 69% to 92%, with remaining variation explained by nonshared environmental effects. Adjustment for body mass index had virtually no effect on the heritability estimates. In this first twin study on lipids to include AAs, no ethnic differences in heritability were found except for LDL, where AAs exhibited higher estimates. PMID- 16212839 TI - Heritability and stability of resting blood pressure. AB - We examined the contribution of genetic and environmental influences to variation in resting systolic (SBP) and diastolic (DBP) blood pressure in participants from 4 twin studies carried out between 1986 and 2003. A total of 1577 subjects (682 males, 895 females) participated. There were 580 monozygotic twins, 664 dizygotic twins and 333 of their siblings. The 4 studies sampled subjects in different age groups (average age 17, 32, 37, 44 years), allowing for comparison of the relative contribution of genetic and environmental factors across the first part of the life span. Blood pressure was assessed under laboratory conditions in 3 studies and by ambulatory monitoring in 1 study. Univariate analyses of SBP and DBP showed significant heritability of blood pressure in all studies (SBP h(2) 48% to 60%, DBP h(2) 34% to 67%). Overall, there was little evidence for sex differences in blood pressure heritability, and no evidence for differences in heritability due to measurement strategy (laboratory vs. ambulatory). For 431 subjects there were data from 2 or more occasions that allowed us to assess the tracking of blood pressure over time and to estimate the genetic and environmental contributions to blood pressure tracking. Correlations over time across an average period of 7.1 years (tracking) were between .41 and .70. Multivariate genetic analyses showed that blood pressure tracking was entirely explained by the same genetic factors being expressed across time. It was concluded that whole genome scans for resting blood pressure can safely pool data from males and females, laboratory and ambulatory recordings, and different age cohorts. PMID- 16212840 TI - Are twins at risk of cancer: results from the Swedish family-cancer database. AB - A few twin studies on cancer have addressed questions on the possible carcinogenic or protective effects of twining by comparing the occurrence of cancer in twins and singletons. The nationwide Swedish Family-Cancer Database of 10.2 million individuals and 69,654 0- to 70-year-old twin pairs were used to calculate standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) for all main cancers compared to singletons. The overall risk of cancer in same- or different-sex twins was at the same level as the risk for singletons. Testicular cancer, particularly seminoma, was increased among same-sex twins (1.54) and all twins to an SIR of 1.38. Among other tumors, neurinomas and non thyroid endocrine gland tumors were increased. Colorectal cancers and leukemia were decreased among all twins. Melanoma and squamous cell skin cancer were decreased in male same-sex twins. The data on this unselected population of twins suggest that twinning per se is not a risk factor of cancer. In utero hormonal exposures or postnatal growth stimulation may be related to the risk of testicular cancer and pituitary tumors. Protective effects against colorectal cancer may be related to a beneficial diet, and in melanoma and skin cancer, to socioeconomic factors. The study involved multiple comparisons, and internal consistency between the results was one of the main factors considered for their plausibility. The results should encourage others working on twin and singleton populations to examine the specific associations and emerging hypotheses. PMID- 16212841 TI - Variations in the maternal age effect on twinning rates: the Nordic experience. AB - Maternal age is the most important nongenetic factor influencing the twinning rate. Every study of the effect of other factors should consider the age distribution of the mothers. Besides standardizing techniques, the age-specific twinning rates are analyzed. Especially, the slope of the regression line is considered when the age-specific twinning rates are regressed against maternal age. How well the maximum age-specific twinning rate measures the variation in the maternal age effect is also studied. The two measures, which are strongly correlated, give consistent results. The proposed methods are applied to data from Finland (1866 to 2003) and Denmark (1855 to 2000). Comparisons with Sweden (1861 to 2000) and Norway (1855 to 2003) are also made. In general, marked decreasing trends in the twinning rates are discernible up to the middle of the 20th century. In Sweden the decline starts around the beginning of the 20th century and is stronger than in Denmark and Norway. The decrease in the twinning rate for Finland, which up to World War II was rather isolated, sets in around the middle of the 20th century and is not as strong as in other Nordic countries. After the minima around 1980, strong increasing trends are noted. This is mainly caused by the influence of the artificial reproduction technologies and particularly the use of fertility-enhancing drugs on the commonly noted dependence between maternal age and the twinning rate. Similar temporal trends can be observed in the slopes of the regression line and in the maximum age specific twinning rates. PMID- 16212842 TI - Risk of infant mortality among twins in relation to placental abruption: contributions of preterm birth and restricted fetal growth. AB - While preterm birth and restricted fetal growth are strongly associated with infant mortality, the extent to which these associations are modified by placental abruption remains unknown. A retrospective cohort study was carried out to examine the risk of infant mortality among twins in relation to abruption, and explore the independent contributions of preterm birth and restricted fetal growth to these associations. The study was restricted to women who had delivered twins at 22 weeks' gestation or more and fetuses weighing 500 grams or more in the United States (1989-2000). Risks of preterm birth (less than 37 weeks' gestation), fetal growth restriction and infant mortality, in relation to placental abruption, were evaluated. All analyses were adjusted for potential sociodemographic confounding factors. The association between restricted twin fetal growth and abruption was the strongest among the most severely growth restricted babies (i.e., less than 1 centile), with the strength of association diminishing with increasing birthweight centiles. The risk of preterm birth among pregnancies with and without abruption were 80.1% and 51.9%, respectively (relative risk [RR] 1.5, 95% confidence interval [CI] 1.4-1.6). The risk of small for-gestational-age (SGA; birthweight of less than the 10th centile for gestational age) among abruption and nonabruption births was 11.7% and 9.2%, respectively (RR 1.3, 95% CI 1.2-1.4). Compared with twins of the appropriate growth delivered at term, the relative risks for infant mortality in the presence of abruption were 9.9 (95% CI 5.4-18.2) for term-SGA, 25.0 (95% CI 22.3-28.1) for preterm-non-SGA, and 36.2 (95% CI 28.4-46.1) for preterm-SGA births. The association between infant mortality and abruption among twins appears largely mediated through preterm birth, and to a lesser extent, through SGA. The association between fetal growth and abruption is strongest among the severely growth-restricted babies, suggesting that the origins of placental abruption may develop in early pregnancy. PMID- 16212843 TI - The Hispanic paradox in twin pregnancies. AB - The objective of this study was to compare length of gestation, fetal growth, and birthweight by race/ethnicity and pregravid weight groups in twin pregnancies. Three thousand and thirty-six twin pregnancies of 28 weeks or more gestation were divided by race/ethnicity (White, Black and Hispanic), and pregravid body mass index (BMI) groups (less than 25.0 vs. 25.0 or more). Outcomes were modeled using multiple regression, controlling for confounders, with White non-Hispanic women as the reference group. Hispanic women had the highest average birthweight and the longest gestation, as well as the lowest proportions of low birthweight, very low birthweight, preterm and early preterm births of the 3 race/ethnicity groups. In the multivariate analyses, Hispanic women had significantly longer gestations (by 7.8 days) and faster rates of fetal growth midgestation (20 to 28 weeks, by 17.4 g/week) and late gestation (after 28 weeks, by 5.3 g/week), whereas Black women had significantly slower rates of fetal growth (by 5.7 g/week and by 4.5 g/week, respectively). These findings in twins reflect the racial and ethnic disparities previously shown in singletons, including the Hispanic paradox of longer gestations and higher rates of fetal growth. PMID- 16212844 TI - Twin transplants; twin study summary; human interest topics. AB - An overview of the nature and success of organ transplantation in monozygotic (MZ) twins is presented. Recent timely examples demonstrate the unique donor recipient relationship shared by MZ co-twins. Next, a recent analysis of MZ co twin differences in the epigenetic profile and associations with differences in life history factors is presented. This work has been the focus of considerable attention by scientists and the media. The final section of this column summarizes information and events concerning twins' longevity, screen careers, older parenting of twins and the origin of the word 'twin'. PMID- 16212846 TI - Profiling mental health needs: what about your Irish patients? PMID- 16212847 TI - Cancer survivorship: a challenge for primary care physicians. PMID- 16212848 TI - The impact of obesity on drug prescribing in primary care. AB - BACKGROUND: Healthcare costs attributable to obesity have previously involved estimations based on costs of diseases commonly considered as having obesity as an underlying factor. AIM: To quantify the impact of obesity on total primary care drug prescribing. DESIGN OF STUDY: Review of computer generated and handwritten prescriptions to determine total prescribing volume for all drug classes. SETTING: Twenty-three general practice surgeries in the UK. METHOD: Stratified random selection of 1150 patients who were obese (body mass index [BMI]>30 kg/m(2)) and 1150 age and sex-matched controls of normal weight (BMI 18.5-<25 kg/m(2)). Retrospective review of medical records over an 18-month period. RESULTS: A higher percentage of patients who were obese, compared with those of normal weight, were prescribed at least one drug in the following disease categories: cardiovascular (36% versus 20%), central nervous system (46% versus 35%), endocrine (26% versus 18%), and musculoskeletal and joint disease (30% versus 22%). All of these categories had a P-value of <0.001. Other categories, such as gastrointestinal (24% versus 18%), infections (42% versus 35%), skin (24% versus 19%) had a P-value of <0.01, while respiratory diseases (18% versus 21%) had a P-value of <0.05. Total prescribing volume was significantly higher for the group with obesity and was increased in the region of two- to fourfold in a wide range of prescribing categories: ulcer healing drugs, lipid regulators, beta-adrenoreceptor drugs, drugs affecting the rennin angiotensin system, calcium channel blockers, antibacterial drugs, sulphonylureas, biguanides, non-steroidal anti-inflammatories (NSAIDs) (P<0.001) and fibrates, angiotensin II antagonists, and thyroid drugs (P<0.05). The main impact on prescribing volumes is from numbers of patients treated, although in some areas there is an effect from greater dosage or longer treatment in those who are obese including calcium channel blockers, antihistamines, hypnotics, drugs used in the treatment of nausea and vertigo, biguanides, and NSAIDs (P<0.05) reflected in significantly increased defined daily dose prescribing. CONCLUSIONS: This large study of contemporary practice indicates that obesity more than doubled prescribing in most drug categories. PMID- 16212849 TI - A qualitative study of GPs' views of treating obesity. AB - BACKGROUND: Due to the increased prevalence of obesity GPs now have a key role in managing obese patients. AIM: To explore GPs' views about treating patients with obesity. SETTING: An inner London primary care trust. DESIGN OF STUDY: A qualitative study using semi-structured interviews. METHOD: Twenty-one GPs working in an inner London primary care trust were interviewed about recent obese patients and obesity in general. An interpretative phenomenological approach was used for data analysis. RESULTS: GPs primarily believed that obesity was the responsibility of the patient, rather than a medical problem requiring a medical solution. They also believed that in contrast to this, obese patients wanted to hand responsibility over to their doctor. This contradiction created conflict for the GPs, which was exacerbated by a sense that existing treatment options were ineffective. Further, this conflict was perceived as potentially detrimental to the doctor-patient relationship. GPs described a range of strategies that they used to maintain a good relationship including offering anti-obesity drugs, in which they had little faith, as a means of meeting patients' expectations; listening to the patients' problems, despite not having a solution to them; and offering an understanding of the problems associated with being overweight. CONCLUSION: GPs believe that although patients want them to take responsibility for their weight problems, obesity is not within the GP's professional domain. Until more effective interventions have been developed GPs may remain unconvinced that obesity is a problem requiring their clinical expertise and may continue to resist any government pressure to accept obesity as part of their workload. PMID- 16212850 TI - A qualitative study exploring how GPs decide to prescribe antidepressants. AB - BACKGROUND: To influence GPs' prescribing policies and practices it is necessary to have an understanding of how they make decisions. The limited evidence available suggests that not only do GPs find making decisions about diagnosing and prescribing for depression problematic, but that decisions are severely constrained by lack of resources. As a result, it might be thought that GPs, in line with current guidelines, will inevitably prescribe antidepressants for patients presenting with symptoms of anxiety and depression. This study examines the accuracy of this view. AIM: To explore how GPs decide to prescribe antidepressants. DESIGN: Focus groups with self-selected GPs. SETTING: Bristol and the surrounding district. METHOD: Qualitative study of five focus groups with 27 GPs. RESULTS: GPs' decisions about whether an antidepressant would be an appropriate form of management are shaped by a set of rules based on 'clinical' and 'social' criteria. The preferred strategy is to 'wait and see', but antidepressants are prescribed earlier when symptoms are perceived to be persistent, unresolving, severe and 'classic'. Decisions to prescribe are also shaped by organisational constraints of time, lack of accessible alternative management options, cost of prescribing and perceived patient attitude. CONCLUSION: The evidence from this study provides little support for the view that GPs take the easy option of prescribing antidepressants in the face of uncertainty. Evidence suggests that the GPs' prescribing was cautious, which indicates that GPs would support the initiative of recent draft guidelines regarding watchful waiting. This guidance, however, needs to be clear about what constitutes mild depression and address the question of prescribing to patients who are experiencing social adversity. Furthermore, alternatives to antidepressants such as counselling would need to be readily and equitably accessible. In addition, GPs need to be convinced that alternatives to antidepressants are at least as effective for patients with so-called 'mild depression'. PMID- 16212851 TI - Treatment of anxiety disorders in primary care practice: a randomised controlled trial. AB - BACKGROUND: Anxiety disorders are prevalent in primary care. Psychological treatment is effective but time-consuming, and there are waiting lists for secondary care. Interest has therefore grown in developing guidelines for treatment that would be feasible in primary care. AIM: To compare the effectiveness and feasibility of guided self-help, the Anxiety Disorder Guidelines of the Netherlands College of General Practitioners and cognitive behavioural therapy (CBT). DESIGN OF STUDY: Randomised controlled study lasting 12 weeks with follow-up at 3 and 9 months for primary care patients with panic disorder and/or generalised anxiety disorder. SETTING: The first two forms of treatment were carried out by 46 GPs who were randomly assigned to one or the other form. CBT was carried out by cognitive behaviour therapists in a psychiatric outpatient clinic. METHOD: Participants (n = 154) were randomly assigned to one of the three forms of treatment. The main outcome measure used was the state subscale of the Spielberger Anxiety Inventory. RESULTS: All three forms of treatment gave significant improvement between pre-test and post-test, and this improvement remained stable between post-test and the follow-ups. The results obtained with the three treatment forms did not differ significantly over time. The feasibility of the Anxiety Disorder Guidelines was low compared with that of guided self-help. CONCLUSION: Our results indicate that primary care patients with prevalent anxiety disorders for whom the GP does not find referral necessary can be adequately treated by the GP. Psychiatric outpatient clinic referral does not give superior results. Guided self-help is easier for the GP to carry out than a less highly-structured treatment like that laid down in the Anxiety Disorder Guidelines. PMID- 16212852 TI - Mental health in the Dutch population and in general practice: 1987-2001. AB - BACKGROUND: In the last 15 years, both the demand for and supply of specialised mental health care increased considerably in the Netherlands. Increased demand may reflect a change in psychological morbidity, but may also be a consequence of increased supply. Specialised health care in the Netherlands is accessible only through referral by a GP, and so it is important to consider the role of primary care in the diagnosis of mental health problems. AIM: The aim of this study is to achieve a better understanding of the development of mental health status in the Dutch population and the consequent help-seeking behaviour in primary care. METHOD: Using two comparable morbidity studies carried out in the Dutch population and in primary care, we compared data from 1987 and 2001 to assess the following: possible differences in mental health between 1987 and 2001; possible differences in prevalence of mental disorder as diagnosed by GPs in 1987 and 2001; possible differences in the sociodemographic determinants of mental health and mental disorder in primary care between 1987 and 2001. RESULTS: Our results show an increase in mental and social problems in the population between 1987 and 2001. However, GPs diagnosed fewer patients as having a mental disorder in 2001 than they did in 1987. The risk of mental disorders or social problems in several sociodemographic groups remained largely the same, as did the chance of receiving a psychological or social diagnosis. CONCLUSION: We conclude that, while mental disorder in the population is increasing, the role of primary care has changed. Although GPs diagnose a lower percentage of mental problems as such, they refer an increasingly larger proportion of these to secondary care. PMID- 16212854 TI - Sharing patient data: competing demands of privacy, trust and research in primary care. AB - BACKGROUND: Patient privacy may conflict with the advancement of knowledge through data sharing. The data contained in primary care records are uniquely comprehensive. AIM: To explore the knowledge and attitudes of patients and members of the primary healthcare team regarding the sharing of data held in primary care records, with particular reference to data sharing for research and the impact that this may have on trust between patients and health professionals. DESIGN OF STUDY: Qualitative study using quota sampled, semi-structured interviews. SETTING: Five general practices in Leicestershire, UK. METHOD: Grounded theory and framework methodology were used. Interviews were transcribed and analysed thematically. RESULTS: Twenty patients and 15 healthcare professionals and managers were interviewed. Patients had limited knowledge of the type of information held in their general practice records and the ways in which these data are shared, but appeared ready to form preliminary views on issues such as data sharing for audit and disease registration. In this climate of limited awareness, there was no suggestion that concern about data sharing for research adversely affects patient trust or leads patients to withhold relevant information from health professionals in primary care. Interviews carried out with staff suggested a lack of clear practice policies regarding data sharing. CONCLUSIONS: General practices may need to develop policies on data sharing, bring these to the attention of their patient population and improve patient awareness about the nature of the data contained in their records. Researchers should ensure that patients are adequately informed about the nature of data contained in patient records when seeking consent for data extraction. PMID- 16212853 TI - Urinary incontinence in older people living in the community: examining help seeking behaviour. AB - BACKGROUND: Only a small proportion of older people with urinary incontinence seek help, despite the availability of adequate treatment. AIM: To ascertain the patient- and disease-specific factors that determine whether medical care for urinary incontinence is sought by independently living older people with urinary incontinence. DESIGN OF STUDY: Qualitative and quantitative analyses of interview data. SETTING: All independently-living older patients aged 60 years or over from nine family practices involved in the Nijmegen Monitoring Project. METHOD: All the independently-living patients aged 60 years or over with uncomplicated urinary incontinence were interviewed at home using the Protection, Amount, Frequency, Adjustment, Body image (PRAFAB) assessment tool, the Incontinence Impact Questionnaire, and the Urogenital Distress Inventory (UDI). RESULTS: In total, 56 men and 314 women were interviewed. Half of the patients had sought help from a GP. Help-seeking was related to the duration of symptoms, the severity of incontinence, the impact experienced emotionally and/or physically, and the presence of concomitant symptoms, particularly of urinary obstruction. Only the presence of incontinence-related complaints as listed on the UDI (odds ratio = 2.74, 95% confidence interval = 1.42 to 5.29) was a significant predictor of help-seeking. Most of the patients who had not sought help did not do so because they considered incontinence as not very serious, or because of a lack of knowledge about cause and treatment options - comments such as 'incontinence is age-related', and 'there is nothing that can be done about incontinence', were reported. Major reasons for seeking help were perceived increase in severity or distress and the need for incontinence materials. CONCLUSIONS: Seeking help is particularly determined by the impact experienced and presence of concomitant symptoms. When patients perceive their incontinence as not very serious or distressing and have a lack of knowledge about cause and treatment options, they usually do not seek help. When they perceive an increase in severity or distress or require incontinence materials, they usually do seek help. PMID- 16212855 TI - The impact of NHS Direct on the demand for out-of-hours primary and emergency care. AB - In order to assess the impact of NHS Direct on out-of-hours primary and emergency care, we sought data on service demand from all GP cooperatives, ambulance services and emergency departments in England, Wales and Scotland. We analysed the impact of NHS Direct on demand, taking advantage of the fact that the service was introduced in waves over a period of 2 years. The results showed that the introduction of NHS Direct was associated with a reduction in calls to GP cooperatives, but with no evident effect on emergency services. PMID- 16212856 TI - Reflections on the doctor-patient relationship: from evidence and experience. PMID- 16212857 TI - The 'bulging fontanelle' to be included in primary care algorithms. PMID- 16212858 TI - All GPs are different and some are more different than others. PMID- 16212859 TI - When the drugs don't work - or do they? PMID- 16212860 TI - Homeopathy - a response. PMID- 16212864 TI - Turbulent times in Italian general practice. PMID- 16212865 TI - Registering organ donor preferences - a third way? PMID- 16212866 TI - Half a day at the movies: film studies in the VTS course. PMID- 16212869 TI - Is an NHS designed around the patient bad for your health? PMID- 16212874 TI - Dynamic and reversibility of heterochromatic gene silencing in human disease. AB - In eukaryotic organisms cellular fate and tissue specific gene expression are regulated by the activity of proteins known as transcription factors that by interacting with specific DNA sequences direct the activation or repression of target genes. The post genomic era has shown that transcription factors are not the unique key regulators of gene expression. Epigenetic mechanisms such as DNA methylation, post-translational modifications of histone proteins, remodeling of nucleosomes and expression of small regulatory RNAs also contribute to regulation of gene expression, determination of cell and tissue specificity and assurance of inheritance of gene expression levels. The relevant contribution of epigenetic mechanisms to a proper cellular function is highlighted by the effects of their deregulation that cooperate with genetic alterations to the development of various diseases and to the establishment and progression of tumors. PMID- 16212875 TI - The human leucocyte differentiation antigens (HLDA) workshops: the evolving role of antibodies in research, diagnosis and therapy. AB - The 8th International Workshop on Human Leucocyte Differentiation Antigens (chaired by Zola H and managed by Swart B) was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8th Workshop. We consider what remains to be achieved (including an estimate of the number of leucocyte surface molecules still to be discovered), and how the field can best move forward. PMID- 16212876 TI - p53-independent pRB degradation contributes to a drug-induced apoptosis in AGS cells. AB - The retinoblastoma (RB) tumor suppressor protein, pRB, plays an important role in the regulation of mammalian cell cycle. Furthermore, several lines of evidence suggest that pRB also involves in the regulation of apoptosis. In the present study, the degradation of pRB was observed in apoptotic gastric tumor cells treated with a new potent anti-tumor component, tripchlorolide (TC). The inhibition of pRB degradation by a general cysteine protease inhibitor IDAM resulted in the reduction of the apoptotic cells. Furthermore, the survival of the gastric tumor cells under the TC treatment was enhanced by an over-expression of exogenous pRB. These results suggest that the pRB degradation of the gastric tumor cells under the TC treatment involves in the apoptotic progression. In addition, the same extent of TC-induced pRB-degradation was detected in the gastric tumor cells containing a p53 dominant-negative construct, indicating that this kind of pRB degradation is p53-independent. PMID- 16212877 TI - Trichomonas vaginalis perturbs the junctional complex in epithelial cells. AB - Trichomonas vaginalis, a protist parasite of the urogenital tract in humans, is the causative agent of trichomonosis, which in recent years have been associated with the cervical cancer development. In the present study we analyzed the modifications at the junctional complex level of Caco-2 cells after interaction with two isolates of T. vaginalis and the influence of the iron concentration present in the parasite's culture medium on the interaction effects. Our results show that T. vaginalis adheres to the epithelial cell causing alterations in the junctional complex, such as: (a) a decrease in transepithelial electrical resistance; (b) alteration in the pattern of junctional complex proteins distribution as observed for E-cadherin, occludin and ZO-1; and (c) enlargement of the spaces between epithelial cells. These effects were dependent on (a) the degree of the parasite virulence isolate, (b) the iron concentration in the culture medium, and (c) the expression of adhesin proteins on the parasite surface. PMID- 16212878 TI - Long-term modifications of blood pressure in normotensive and spontaneously hypertensive rats by gene delivery of rAAV-mediated cytochrome P450 arachidonic acid hydroxylase. AB - Arachidonic acid cytochrome P-450 (CYP) hydroxylase 4A isoforms, including 4A1, 4A2, 4A3 and 4A8 in the rat kidney, catalyze arachidonic acid to produce 19/20 Hydroxyeicosatetraenoic acids (20-HETE), a biologically active metabolite, which plays an important role in the regulation of blood pressure. However, controversial results have been reported regarding the exact role of 20-HETE on blood pressure. In the present study, we used recombinant adeno-associated viral vector (rAAV) to deliver CYP 4A1 cDNA and antisense 4A1 cDNA into Sprague-Dawley (SD) rats and spontaneously hypertensive rats (SHR), respectively, to investigate the effects of long-term modifications of blood pressure and the potential for gene therapy of hypertension. The mean systolic pressure increased by 14.2+/-2.5 mm Hg in rAAV.4A1-treated SD rats and decreased by 13.7+/-2.2 mm Hg in rAAV.anti4A1-treated SHR rats 5 weeks after the injection compared with controls and these changes in blood pressure were maintained until the experiments ended at 24 weeks. In 4A1 treated animals CYP4A was overexpressed in various tissues, but preferentially in the kidney at both mRNA and protein levels. In anti-4A1 treated SHR, CYP4A mRNA in various tissues was probed, especially in kidneys, but 4A1 protein expression was almost completely inhibited. These results suggest that arachidonic acid CYP hydroxylases contribute not only to the maintenance of normal blood pressure but also to the development of hypertension. rAAV-mediated anti4A administration strategy has the potential to be used as targeted gene therapy in human hypertension by blocking expression of CYP 4A in kidneys. PMID- 16212879 TI - AtKP1, a kinesin-like protein, mainly localizes to mitochondria in Arabidopsis thaliana. AB - Kinesins and kinesin-like proteins (KLPs) constitute a large family of microtubule-based motors that play important roles in many fundamental cellular and developmental processes. To date, a number of kinesins or KLPs have been identified in plants including Arabidopsis thaliana. Here, a polyclonal antibody against AtKP1 (kinesin-like protein 1 in A. thaliana) was raised by injection the expressed AtKP1 specific C-terminal polypeptides in rabbits, and immunoblot analysis was conducted with the affinity-purified anti-AtKP1 antibody. The results indicated that this antibody recognized the AtKP1 fusion proteins expressed in E. coli and proteins of ~125 kDa in the soluble fractions of Arabidopsis extracts. The molecular weight was consistent with the calculated molecular weight based on deduced amino acids sequence of AtKP1. To acquire the subcellular localization of the protein, AtKP1 in Arabidopsis root cells was observed by indirect immunofluorescence microscopy. AtKP1 was localized to particle-like organelles in interphase or dividing cells, but not to mitotic microtubule arrays. Relatively more AtKP1 was found in isolated mitochondria fraction on immunoblot of the subcellular fractions. The AtKP1 protein could not be released following a 0.6 M KI washing, indicating that AtKP1 is tightly bind to mitochondria and might function associated with this kind of organelles. PMID- 16212880 TI - A novel in vitro system for gamete fusion in maize. AB - Various systems by using electric pulse, calcium, or polyethylene glycol have been developed in the past decade for the in vitro fusion of plant gametes. These in vitro systems provide a new way to study the fertilization mechanisms of plants. In this study, we developed a bovine serum albumin (BSA)-mediated fusion system for the in vitro fusion of maize gametes. The in vitro fusion of the isolated single egg cell and sperm cell of maize was observed microscopically in the BSA solution and the fertilized egg cell showed normal cell wall regeneration and nuclear division. The effects of the BSA concentration, pH value and calcium level on the efficiency of the maize gamete fusion were also assessed. BSA concentration and pH value did significantly affect the efficiency of the gamete fusion. Calcium was not necessary for the gamete fusion when BSA was present. The optimal solution for the gamete fusion contained 0.1% BSA, pH 6.0. The fusion frequency was as high as 96.7% in that optimal solution. This new in vitro fertilization system offers an alternative tool for the in vitro study of fertilization mechanisms with much simpler manipulating procedure than PEG system, and it will be especially useful for the in vitro study of the calcium dynamics during plant fertilization. PMID- 16212881 TI - Generation and characterization of novel stromal specific antibodies. AB - Rheumatoid synovial fibroblasts were used as an immunogen to produce monoclonal antibodies selected for their reactivity with stromal cell antigens. Mice were immunised with low passage whole cell preparations and the subsequent hybridomas were screened by immunohistochemistry on rheumatoid synovium and tonsil sections. The aim was to identify those antibodies that recognised antigens that were restricted to stromal cells and were not expressed on CD45 positive leucocytes. A significant number of antibodies detected antigen that identified endothelial cells. These antibodies were further characterised to determine whether the vessels identified by these antibodies were vascular or lymphatic. From five fusions clones were identified with predominant reactivity with: 1) fibroblasts and endothelial cells; or 2) broad stromal elements (fibroblast, endothelium, epithelium, follicular dendritic cells). A fibroblast-specific antibody that did not also identify vessels was not generated. Examples of each reactivity pattern are discussed. PMID- 16212882 TI - The Qa-1 dependent CD8+ T cell mediated regulatory pathway. AB - The immune system has evolved a variety of regulatory mechanisms to ensure the peripheral self-tolerance as well as the optimal capacity to elicit effective anti-infection immunity. At present, there is no satisfactory conceptual framework to explain how the peripheral immunity is regulated at a biological system level, which enables the immune system to perform its essential functions to mount effective immunity to virtually any foreign antigens but avoid harmful immune responses to self. In this regard, during the past few years, an "affinity/avidity model of peripheral T cell regulation" has been proposed and tested, which opens up a new paradigm to understand how the peripheral immunity, to both self and foreign antigens, is regulated. The paradigm is based on the discovery of a subset CD8+ T cells with TCRs which specifically recognize a unique set of self-peptides presented by the MHC class Ib molecule Qa-1 differentially expressed on T cells as a function of the affinity/avidity of T cell activation. These Qa-1 restricted CD8+ T cells represent an example of how the immune system utilizes a unified mechanism to regulate adaptive immunity to both self and foreign antigens. Thus, by selectively down-regulating T cells of intermediate affinity/avidity, to any antigens, the immune system controls the adaptive immunity without the necessity to distinguish self from non-self in the periphery at the level of T cell regulation. PMID- 16212883 TI - Antigen processing by autoreactive B cells promotes determinant spreading. AB - Acute primary immune responses tend to focus on few immunodominant determinants using a very limited number of T cell clones for expansion, whereas chronic inflammatory responses generally recruit a large number of different T cell clones to attack a broader range of determinants of the invading pathogens or the inflamed tissues. In T cell-mediated organ-specific autoimmune disease, a transition from the acute to the chronic phase contributes to pathogenesis, and the broadening process is called determinant spreading. The cellular components catalyzing the spreading reaction are not identified. It has been suggested that autoreactive B cells may play a central role in diversifying autoreactive T cell responses, possibly through affecting antigen processing and presentation. The clonal identity and diversity of the B cells and antibodies seem critical in regulating T cell activity and subsequent tissue damage or repair. Here, we use two autoimmune animal models, experimental autoimmune thyroiditis (EAT) and type 1 diabetes (T1D), to discuss how autoreactive B cells or antibodies alter the processing and presentation of autoantigens to regulate specific T cell response. PMID- 16212884 TI - Molecular characteristics of cockroach allergens. AB - Cockroaches, commonly found in urban dwellings worldwide, have long been considered vectors of various infectious diseases and cockroach allergens are one of the major etiologic risk factors for IgE-mediated allergic respiratory illness throughout the world. A high prevalence of cockroach hypersensitivity in atopic (20-55%) and asthmatic (49-60%) populations has been documented. Cockroach allergens with molecular weights ranging from 6 to 120 kD have been identified by various standard immunochemical techniques. This article covers the characteristics of major cockroach allergens that have been purified, sequenced, cloned, and produced as recombinant proteins. PMID- 16212885 TI - Immunomodulatory effects of esculetin (6,7-dihydroxycoumarin) on murine lymphocytes and peritoneal macrophages. AB - Coumarins belong to a diverse group of naturally occurring non-nutrient phytochemicals known as benzo-alpha-pyrones. In this study, esculetin, a 6,7 dihydroxy derivative of coumarin with pleiotropic biological activities, was found to have no significant cytotoxic effect on normal murine macrophages, but it could increase the in vivo migration of the thioglycollate-elicited macrophages in a dose-dependent manner. Moreover, esculetin significantly increased the endocytic activity, and augmented the nitric oxide production and iNOS gene expression in LPS-treated macrophages. In addition, in vivo administration of esculetin into mice was shown to increase the mitogenesis of splenic lymphocytes towards Con A and LPS stimulations, and induced the LAK activity of splenic lymphocytes. Collectively, our results indicate that esculetin could exert immunomodulatory effects on murine macrophages and lymphocytes, both in vitro and in vivo, and this might be one of the possible mechanisms by which coumarins can exert their chemopreventive and anti-tumor activities in vivo. PMID- 16212886 TI - Antisense-induced blockade of GATA-3 expression could inhibit Th2 excursion of tumor cells in vitro and in vivo. AB - Previous studies have shown that tumor cells predominantly express Th2 type cytokines and transcription factors. GATA-3, as a Th2-specific transcription factor, plays a central role in positive-regulating Th2 development. So whether the expression of GATA-3 in tumor cells has any effect on tumor development is a question of interest. In the present study, we inhibited the expression of GATA-3 in tumor cells through antisense RNA blockade technique, and observed its effects on tumor in vitro and in vivo. Our results showed that antisense GATA-3 treatment could inhibit the expression of TNF-alpha and Th2 cytokines in tumor cells, and antisense-induced blockade of GATA-3 could also depress tumor growth in tumor bearing mice. We suggest that the ratio of T-bet/GATA-3 can be evaluated as a more important marker of the status of Th1/Th2 type. And our results might provide some evidence about the molecular regulatory mechanisms in tumor cell development. PMID- 16212887 TI - Effect of protein kinase C on proliferation and apoptosis of T lymphocytes in idiopathic thrombocytopenic purpura children. AB - It is well-documented that T lymphocyte proliferation and apoptosis are abnormal in idiopathic thrombocytopenic purpura (ITP) children. However, the underlying regulation mechanisms especially in terms of signal transduction remain unknown. In this paper, we reported the changes of protein kinase C (PKC) activity in peripheral blood T lymphocytes and the effect of PKC on T lymphocyte proliferation and apoptosis. We demonstrated that in ITP children, the activator (PMA) and inhibitor (H-7) of PKC affected on T lymphocyte proliferation and apoptosis dramatically, but they altered little in healthy children. PKC activity was significantly enhanced in ITP children together with an increased expression of FasL on CD3+T, CD4+T and CD8+T cells, resulting in a positive correlation between PKC activity and the expression of FasL on T cells. While the PKC activity and the platelet count were negatively correlated. Taken together, our findings suggest that the PKC activation may enhance T lymphocytes activity, suppress T cell apoptosis and be involve in thrombocytes damage as a mechanism related to immune pathogenesis of ITP. PMID- 16212888 TI - The distribution of synaptotagmin II in RBL-2H3 and its regulation on exocytosis of lysosomes in RBL-2H3. AB - Synaptotagmin (Syt) constitutes a family of membrane-trafficking proteins, so far nearly 20 Syts have been discovered. Extensive work showed that synatotagmins were a potential Ca2+ sensor for regulated exocytosis. This study was to investigate the expression and location of synaptotagmin II (Syt2) in RBL-2H3 (RBL) and its role in regulating exocytosis of RBL. The expression of Syt2 in RBL was confirmed by Western blot. The recombinant expression vector pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation, the stable transfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated at plasma membrane with little detected in cytoplasm. To analyze the role of Syt2 during exocytosis of RBL, the release of cathepsin D was assayed by immunoblotting. Compared with control, the release of cathepsin D by RBL-Syt2-S was markedly decreased. The results indicated that Syt2 played a negative regulation in exocytosis of lysosomes in RBL. PMID- 16212889 TI - STAT1 antisense oligonucleotides attenuate the proinflammatory cytokine release of alveolar macrophages in bleomycin-induced fibrosis. AB - To investigate the effect of signal transducers and activators of transcription 1 (STAT1) antisense oligonucleotides (ASON) on concentrations of TNF-alpha, IL-8, NO secreted by alveolar macrophages (AMs) in bleomycin-induced rat pulmonary fibrosis, five adult female Wistar rats were intratracheally instilled with bleomycin. After 7 days, the rats were killed by right ventricle of heart exsanguinations under ketamine anaesthesia and bronchoalveolar lavage (BAL) was performed to obtain AMs. AMs were divided into four groups, treated with STAT1 ASON, STAT1 sense oligonucleotides (SON), dexamethasone (DEX) and medium alone (control), respectively. AMs and media were collected after culture for 36 h. The mRNA and protein expressions of STAT1 and ICAM-1 in AMs were detected by RT-PCR and ELISA, respectively. The concentrations of TNF-alpha, IL-8, NO in cultured medium were detected. The STAT1 mRNA expression by AMs in the STAT1 ASON group was lower than those of AMs in the STAT1 SON group, the DEX group and the control group (p<0.05). Moreover, the STAT1 mRNA expression by AMs in the DEX group was also lower than those of AMs in the STAT1 SON group and the control group (p<0.05), but the STAT1 mRNA expression by AMs in the STAT1 SON group was not different from that of the control group (p>0.05). The protein expressions of STAT1 and ICAM-1 and the mRNA expression of ICAM-1 showed similar changes to the STAT1 mRNA expression by AMs. The concentrations of TNF-alpha, IL-8, NO in cultured medium from STAT1 ASON group were lower than those from STAT1 SON, DEX and the control groups (p<0.05). Moreover, the concentrations of TNF-alpha, IL-8, NO in cultured medium from DEX group were also lower than those from the control and STAT1 SON group (p<0.05), but no difference between STAT1 SON group and the control (p>0.05). The results suggest that STAT1 ASON could inhibit the secretion of TNF-alpha, IL-8, NO in AMs, and STAT1 could become a target of treating pulmonary fibrosis. PMID- 16212890 TI - Identification and characterization of peptides mimicking the epitopes of metalloprotease of Schistosoma japonicum. AB - In an attempt to isolate and characterize peptides mimicking epitopes of metalloprotease and explore their immunological protection against Schistosoma japonicum (S. japonicum), polyclonal anti-metalloprotease sera was prepared to screen a 12-mer random peptide library to isolate phages binding specially to antisera IgG. Then, phage ELISA, animal immunization, DNA sequencing, Western blotting and enzymatic activity neutralizing analysis were used to characterize the selected phage clones. All of ten randomly picked clones were shown to be positive. Five peptides of different amino acid sequences deduced from DNA sequences were obtained and two of them (peptides 2 and 3) could induce significant reduction (31.0% and 31.8%, respectively) in worm burden and high reduction (52.6% and 54.9%, respectively) in liver eggs per gram (LEPG), while, unexpectedly, others (peptides 1, 4 and 5) could not elicit enough protection against infection of S. japonicum. Peptides 2 and 3 could be recognized by S. japonicum infected mouse sera (IMS) and could elicit neutralizing Abs. The results show that peptides 2 and 3 are antigenic and immunogenic. They are true mimics of epitopes of metalloprotease and useful as novel vaccine candidates against S. japonicum. PMID- 16212891 TI - Differentiation of human bone marrow stromal cells into neural-like cells induced by sodium ferulate in vitro. AB - Human marrow stromal cells (hMSCs) are multipotential stem cells, capable of differentiating into bone, cartilage, fat and muscle. Several recent reports demonstrated that hMSCs have been also differentiated into neural cells. However, only a few reported inducers are applicable for clinical use. This work is to explore the effects of sodium ferulate (SF) on differentiation of hMSCs into neural cells in vitro. We found that hMSCs could be induced to the cells with typical neural morphology when cultured with SF. The cells express neural proteins, such as nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP). About 30% of the hMSC-derived cells expressed nestin when cultured with SF for 3 h, but no expression was detected after 24 h. The percentages of positive cells for NSE or GFAP were about 67% and 39% separately at 6 h, and reached the plateau phage after treatment with SF for 3 days. The data suggest that SF can induce hMSCs to differentiate into neural-like cells in vitro. PMID- 16212892 TI - Preparation of ChIL-2 and IBDV VP2 fusion protein by baculovirus expression system. AB - This study aims to produce an effective subunit vaccine against infectious bursal disease virus (IBDV). The genes of chicken interleukin-2 (ChIL-2) and IBDV viral protein 2 (VP2) were amplified and fused by splice overlap extension-polymerase chain reaction (SOE-PCR). The fusion gene was digested by EcoR I/Kpn I and inserted into pBacPAK8 vector, resulting in recombinant transfer plasmid pBacPakVP2-IL2. The recombinant plasmid was transfected into Sf-9 cells accompanied with hybrid nuclear polyhedrosis virus (HyNPV) genome DNA and lipofectin. Plaque-purification indicated that we had got the recombinant Hy-VP2 IL2. Fusion protein VP2-IL2 was expressed effectively both in insect cells and bombyx mori. The expression of fusion protein was confirmed by ELISA, SDS-PAGE and Western blotting assay, respectively. This efficient system allows us to meet the need for inexpensive vaccines required by the poultry industry. PMID- 16212893 TI - CD8: adhesion molecule, co-receptor and immuno-modulator. AB - CD8 is a cell surface glycoprotein found in cytotoxic T lymphocytes, which are important components in cellular immunity, esp. in the immune response to cancer and chronic infections. There are two forms of CD8, either as an alphaalpha homodimer or alphabeta heterodimer. It acts as an "assistant" or co-receptor in the function of cytotoxic T cells where specific immunity is mediated by interaction of specific T cell receptor (alphabeta TCR) and its ligand peptide major histocompatibility complex (pMHC). CD8 also binds to pMHC but away from the interface of pMHC and TCR contact, thereof no influence on the specificity of this interaction. If the TCR and CD8 bind to the same pMHC at the same time, CD8 is defined as a co-receptor, functioning through its signalling via its cytoplasmic tyrosine phosphorylation pathway; if CD8 binds to pMHC independently of the TCR, it is defined as an adhesion molecule. At present, the co-receptor function theory is dominated in the field. Recent study has also shown that murine CD8 alphaalpha binds to TL antigen, an MHC homologue, therefore acts as an immuno-modulator. In this review, we discuss these current understandings of the three aspects of the CD8 functions and their structural basis. PMID- 16212894 TI - The pre-B cell receptor and its function during B cell development. AB - The process of B cell development in the bone marrow occurs by the stepwise rearrangements of the V, D, and J segments of the Ig H and L chain gene loci. During early B cell genesis, productive Ig H chain gene rearrangement leads to assembly of the pre-B cell receptor (pre-BCR), which acts as an important checkpoint at the pro-B/preB transitional stage. The pre-BCR, transiently expressed by developing precursor B cells, comprises the Ig muH chain, surrogate light (SL) chains VpreB and lambda5, as well as the signal-transducing heterodimer Ig alpha/Ig beta. Signaling through the pre-BCR regulates allelic exclusion at the Ig H locus, stimulates cell proliferation, and induces differentiation to small post-mitotic pre-B cells that further undergo the rearrangement of the Ig L chain genes. Recent advances in elucidating the key roles of pre-BCR in B cell development have provided a better understanding of normal B lymphopoiesis and its dysregulated state leading to B cell neoplasia. PMID- 16212895 TI - Chemokines and chemokine receptors: their manifold roles in homeostasis and disease. AB - Chemokines are a superfamily of small proteins that bind to G protein-coupled receptors on target cells and were originally discovered as mediators of directional migration of immune cells to sites of inflammation and injury. In recent years, it has become clear that the function of chemokines extends well beyond the role in leukocyte chemotaxis. They participate in organ development, angiogenesis/angiostasis, leukocyte trafficking and homing, tumorigenesis and metastasis, as well as in immune responses to microbial infection. Therefore, chemokines and their receptors are important targets for modulation of host responses in pathophysiological conditions and for therapeutic intervention of human diseases. PMID- 16212896 TI - Gene therapy of cancer: induction of anti-tumor immunity. AB - Many malignancies lack satisfactory treatment and new therapeutic options are urgently needed. Gene therapy is a new modality to treat both inherited and acquired diseases based on the transfer of genetic material to the tissues. Different gene therapy strategies against cancers have been developed. A considerable number of preclinical studies indicate that a great variety of cancers are amenable to gene therapy. Among these strategies, induction of anti tumor immunity is the most promising approach. Gene therapy with cytokines has reached unprecedented success in preclinical models of cancer. Synergistic rather than additive effects have been demonstrated by combination of gene transfer of cytokines/chemokines, costimulatory molecules or adoptive cell therapy. Recent progress in vector technology and in imaging techniques allowing in vivo assessment of gene expression will facilitate the development of clinical applications of gene therapy, a procedure which may have a notorious impact in the management of cancers lacking effective treatment. PMID- 16212897 TI - Development of dendritic cell system. AB - The dendritic cell system contains conventional dendritic cells (DCs) and plasmacytoid pre-dendritic cells (pDCs). Both DCs and pDCs are bone marrow derived cells. Although the common functions of DCs are antigen-processing and T lymphocyte activation, they differ in surface markers, migratory patterns, and cytokine output. These differences can determine the fate of the T cells they activate. Several subsets of mature DCs have been described in both mouse and human and the developmental processes of these specialized DC subsets have been studied extensively. The original concept that all DCs were of myeloid origin was questioned by several recent studies, which demonstrated that in addition to the DCs derived from myeloid precursors, some DCs could also be efficiently generated from lymphoid-restricted precursors. Moreover, it has been shown recently that both conventional DCs and pDCs can be generated by the Flt3 expressing hemopoietic progenitors regardless of their myeloid- or lymphoid-origin. These findings suggest an early developmental flexibility of precursors for DCs and pDCs. This review summarizes some recent observations on the development of DC system in both human and mouse. PMID- 16212898 TI - Immunological responses against SARS-coronavirus infection in humans. AB - Since the outbreak of a SARS epidemic last year, significant advances have been made on our understanding of the mechanisms of interaction between the SARS coronavirus (CoV) and the immune system. Strong humoral responses have been found in most patients following SARS-CoV infection, with high titers of neutralizing Abs present in their convalescent sera. The nucleocapsid (N) and spike (S) proteins of SARS-CoV appear to be the dominant antigens recognized by serum Abs. CD4+ T cell responses against the N protein have been observed in SARS patients and an HLA-A2-restricted cytotoxic T lymphocyte epitope in the S protein has been identified. It is likely that the immune responses induced by SARS-CoV infection could also cause pathological damage to the host, especially in the case of proinflammatory cytokines. There is also evidence suggesting that SARS-CoV might be able to directly invade cells of the immune system. Our understanding on the interaction between SARS-CoV, the immune system and local tissues is essential to future diagnosis, control and treatment of this very contagious disease. PMID- 16212899 TI - IL-2 and IL-15 exhibit opposing effects on Fas mediated apoptosis. AB - It has been shown that IL-2 and IL-15 can have opposing effects on life and death of T cells. However, the role of IL-2 and IL-15 in regulating the fate of other cell types is less clear. In the present study, we examined the impact of IL-2 and IL-15 on life and death of pre-B cells using the BAF-B03 line. We showed that BAF-B03 cells constitutively expressed the private IL-2R alpha chain and IL-15R alpha chain, and the shared IL-2R beta chain and gamma c chain. Stimulation of BAF-B03 cells in vitro with IL-2 and IL-15 induced vigorous cell proliferation in a dose-dependent fashion. Titration of IL-2 and IL-15 in the assay showed that the mitotic effects of IL-2 and IL-15 were remarkably similar. However, the sensitivities of BAF-B03 cells to Fas mediated apoptosis after IL-2 and IL-15 stimulation were strikingly different. Cells cultured in IL-2 readily underwent apoptotic cell death upon cross-linking of the Fas receptor whereas cells cultured in IL-15 were extremely resistant to Fas triggered cell death. The anti apoptotic effect of IL-15 in this model was associated with increased expression of Bcl-xL. FLIP expression, however, was comparable between IL-2 and IL-15 stimulated cells. We conclude that IL-2 and IL-15 have diametrically opposite effect on the fate of BAF-B03 cells, although both cytokines share similar receptor structure and exhibit similar mitotic activities. PMID- 16212900 TI - Human prolactin improves engraftment and reconstitution of human peripheral blood lymphocytes in SCID mice. AB - Recombinant human prolactin (rhPRL) was administered to huPBL-SCID mice to determine its effects on human immunologic reconstitution and function. The huPBL SCID mice were given 10 microg i.p. injection of rhPRL every other day for a total of 10 injections after huPBL were transferred. The results demonstrated that rhPRL improved the engraftment of lymphocytes into thymus, lymph nodes and spleens, showing that the cellularities of these organs increased although the cellularities tended to vary depending on the donor. The amounts of human T cells (HLA-ABC+/CD3+) increased greatly in thymus (14.2 folds), spleen (4.16 folds) and lymph nodes (40.18 folds) after rhPRL injections. The amounts of human B cells (HLA-ABC+/CD19+) also increased greatly in lymph nodes (42.5 folds) and spleen (5.78 folds). The lymph node cells from the rhPRL-treated huPBL-SCID mice were more sensitive to PHA stimulation ([3H] thymidine incorporation). The supernatant of PHA-stimulated PBL from rhPRL-treated huPBL/SCID chimerism contained more cytokines (IFN-gamma and IL-2). The natural cytotoxicity against human sensitive target cells, K562 cells, from spleen and bone marrow of hPBL/SCID chimerism was significantly enhanced by rhPRL administration. The lymph node cells were stimulated with LPS in vitro for 3 days and the lymphocytes from the rhPRL treated huPBL-SCID mice were more sensitive to mitogen stimulation. Both serum total IgG level and IgM level of rhPRL-treated huPBL/SCID chimerism were increased, and even without DT-rechallenge the base line of DT-specific IgG was elevated after rhPRL treatment in huPBL-SCID mice. Thus, rhPRL stimulation promotes reconstitution of human immune system in huPBL-SCID mice. PMID- 16212902 TI - Distinct overexpression of Fas ligand on T lymphocytes in aplastic anemia. AB - Increased expression of Fas by hematopoietic progenitors in aplastic anemia (AA) suggests that Fas/Fas ligand (FasL) system plays a key role in the formation of severe pancytopenia. To further confirm the above hypothesis, T cells from 8 patients with AA were systematically studied for their FasL's distribution pattern, releasing manner and proapoptotic activity, compared with normal resting T cells and artificially activated T cell blasts. The results demonstrated that AA T cells abnormally expressed low levels of membrane-bound FasL and contained high levels of intracellular FasL which could be triggered to release by high dose phytohemagglutinin (PHA) pulse-stimulation. The supernatants from the PHA stimulated AA T cells had apparent cytotoxicity against FasL-sensitive Jurkat cells, which could be significantly inhibited by monoclonal antibody against FasL in a dose-dependent manner, or nearly completely abrogated by ultracentrifugation. The above phenomena also appeared on artificially activated T cell blasts, but this was not the case on normal resting T cells. These results indicate that AA T cell is a type of "preactivated" T lymphocyte, characterized by overexpression of FasL, especially intracellular FasL which can be stimulated to release in bioactive exosomes-bound form. Taken together, our data provide further and direct evidence for the hypothesis that T cells might mediate the destruction of hematopoietic progenitor in AA through Fas/FasL system. PMID- 16212901 TI - Expression of intracellular domain of epidermal growth factor receptor and generation of its monoclonal antibody. AB - To prepare monoclonal antibody specific to epidermal growth factor receptor (EGFR) intracellular domain, its gene was amplified from total RNA of A431 cell by RT-PCR. Then the gene was cloned into prokaryotic vector pET30a(+). The recombinant plasmid was transformed into E. coli BL21 (DE3) strain for protein expression. Recombinant protein was induced with IPTG and purified using Ni2+-NTA agarose. Then the anti-EGFR monoclonal antibody (mAb) was prepared with classical hybridoma technique. Positive clones were selected using indirect enzyme-linked immunoabsorbent assay (ELISA). Totally 4 hybridoma clones were obtained and these mAbs were IgG1 (3 clones) and IgG2a (1 clone), respectively. Their light chains were all kappa chains. Western blotting analysis and confocal immunofluorescence assays demonstrated that mAbs could specifically recognize EGFR expressing on A431 carcinoma cell line. The mAbs will be useful in the study of EGFR-mediated signal transduction. PMID- 16212903 TI - CpG ODN enhances immunization effects of hepatitis B vaccine in aged mice. AB - Oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides in contexts of unique sequence (CpG motifs) is active as adjuvant in induction of cellular and humoral immune responses in young mice. To date, there are only limited reports about effect of CpG ODN on immune responses against hepatitis B (HB) infection in aged mice. Our studies demonstrated there were significant increases in secreting of total anti-HB IgG, IgG1 and IgG2a, as well as of IL-12 and IFN gamma, when CpG ODNs were injected together with hepatitis B antigen in aged mice. Moreover, CpG ODN could stimulate proliferation of spleen lymphocytes in a dose-dependent manner. Taken together, the results we obtained indicate that the adding of CpG ODN into the vaccine antigen might be useful in development of more effective vaccination for inducing anti-HB virus responses in the elderly. PMID- 16212904 TI - The inhibitory effects of mouse ICOS-Ig gene-modified mouse dendritic cells on T cells. AB - The main approach to reduce graft rejection has been focused on the development of immunosuppressive agents at present. Although these strategies have reportedly reduced graft rejection, there has been a reciprocal increase in more severe immunosuppression and lethal infections, as well as severe side effects. Blockade of costimulatory T cell response has been proved as one of useful strategies to reduce graft rejection. Furthermore, it has been shown that infusion of dendritic cells (DCs) with a potent negative regulatory ability for T cells could prolong allograft survival. In this study mouse DCs (mDCs) were transfected with the recombinant plasmid pcDNA3.0 containing mouse inducible costimulator-Ig (mICOS Ig) cDNA by electroporation. The transient expression of mICOS-Ig in mDC could be detected by ELISA and SDS-PAGE. Mouse ICOS-Ig fusion protein expressed in mDC and mICOS-Ig gene-modified mDC could inhibit lymphocyte proliferation in mixed lymphocyte culture (MLC) in vitro. Furthermore, mICOS-Ig gene-modified mDC could inhibit lymphocyte proliferation in recipient mice. These results suggested that mICOS-Ig gene-modified mDC exerted inhibitory effects on T cells, and might be suitable for treatment or prevention of graft rejection and immunopathologic diseases. PMID- 16212905 TI - Homeostasis of T cell diversity. AB - T cell homeostasis commonly refers to the maintenance of relatively stable T cell numbers in the peripheral lymphoid organs. Among the large numbers of T cells in the periphery, T cells exhibit structural diversity, i.e., the expression of a diverse repertoire of T cell receptors (TCRs), and functional diversity, i.e., the presence of T cells at naive, effector, and memory developmental stages. Although the homeostasis of T cell numbers has been extensively studied, investigation of the mechanisms underlying the maintenance of structural and functional diversity of T cells is still at an early stage. The fundamental feature throughout T cell development is the interaction between the TCR and either self or foreign peptides in association with MHC molecules. In this review, we present evidence showing that homeostasis of T cell number and diversity is mediated through competition for limiting resources. The number of T cells is maintained through competition for limiting cytokines, whereas the diversity of T cells is maintained by competition for self-peptide-MHC complexes. In other words, diversity of the self-peptide repertoire limits the structural (TCR) diversity of a T cell population. We speculate that cognate low affinity self-peptides, acting as weak agonists and antagonists, regulate the homeostasis of T cell diversity whereas non-cognate or null peptides which are extremely abundant for any given TCR, may contribute to the homeostasis of T cell number by providing survival signals. Moreover, self-peptides and cytokines may form specialized niches for the regulation of T cell homeostasis. PMID- 16212906 TI - Specific control of immunity by regulatory CD8 T cells. AB - T lymphocytes with dedicated suppressor function (Treg) play a crucial role in the homeostatic control of immunity in the periphery. Several Treg phenotypes have now been identified in the CD4 and CD8 T cell populations, suggesting their down-regulatory function in both human and animal models of autoimmunity, transplantation and tumor immunity. Here we will focus on the CD8 Treg population and their ability to specifically inhibit a pathogenic autoimmune response. This review will detail the current advances in the knowledge of CD8 Treg in the context of antigen specificity, phenotype, MHC restriction, mechanism of action, and priming. PMID- 16212907 TI - MAP kinases in immune responses. AB - MAP kinases are evolutionarily conserved signaling regulators from budding yeast to mammals and play essential roles in both innate and adaptive immune responses. There are three main families of MAPKs in mammals. Each of them has its own activators, inactivators, substrates and scaffolds, which altogether form a fine signaling network in response to different extracellular or intracellular stimulation. In this review, we summarize recent advances in understanding of the regulation of MAP kinases and the roles of MAP kinases in innate and adaptive immune responses. PMID- 16212908 TI - Dendritic cells in vivo and in vitro. AB - Dendritic cells (DC) are crucial cells of the immune system, and bridge the essential connection between innate and adaptive immunity. They reside in the periphery as sentinels where they take up antigens. Upon activation, they migrate to lymphoid organs and present there the processed antigens to T cells, thereby activating them and eliciting a potent immune response. Dendritic cells are bone marrow-derived cells, still big controversies exist about their in vivo development. In vitro, DC can be generated from multiple precursor cells, among them lymphoid and myeloid committed progenitors. Although it remains unknown how DC are generated in vivo, studying the functions of in vitro generated DC results in fundamental knowledge of the DC biology with promising applications for future medicine. Therefore, in this review, we present current protocols for the generation of DC from precursors in vitro. We will do this for the mouse system, where most research occurs and for the human system, where research concentrates on implementing DC biology in disease treatments. PMID- 16212909 TI - Novel role and regulation of the interleukin-1 receptor associated kinase (IRAK) family proteins. AB - The interleukin-1 receptor associated kinases (IRAKs) sit at the bottle neck for the Toll-like-receptor (TLR) mediated signal transduction process controlling host innate immune response. However, the exact role and regulation of IRAKs are still in the early stage and not fully understood. This review intends to summarize the recent advancement in this important topic and point out areas that need further intensive investigation. PMID- 16212910 TI - Hepatitis B virus X protein sensitizes primary mouse hepatocytes to ethanol- and TNF-alpha-induced apoptosis by a caspase-3-dependent mechanism. AB - It is well-documented that alcohol drinking together with hepatitis viral infection accelerates liver injury; however the underlying mechanisms remain unknown. In this paper, we demonstrated that primary hepatocytes from transgenic mice overexpressing hepatitis B virus X protein (HBX) were more susceptible to ethanol- and TNF-alpha-induced apoptotic killing. Compared to normal control mouse hepatocytes, ethanol and/or TNF-alpha treatment led to a significant increase in reactive oxygen species, mitochondrial permeability transition, cytochrome C release, caspase-3 activity, and poly (ADP-ribose) polymerase degradation in hepatocytes from HBX transgenic mice. Blocking caspase-3 activity antagonized ethanol- and TNF-alpha-induced apoptosis in primary hepatocytes from HBX transgenic mice. Taken together, our findings suggest that HBX sensitizes primary mouse hepatocytes to ethanol- and TNF-alpha-induced apoptosis by a caspase-3-dependent mechanism, which may partly explain the synergistic effects of alcohol consumption and hepatitis B virus infection on liver injury. PMID- 16212911 TI - Selection of proteins for human MHC class II presentation. AB - We investigated the predicted function of proteins eluded from human MHC class II molecules. Peptides that are presented by MHC class II were obtained from the SYFPEITHI database and the corresponding proteins were found in the SWISSPROT database. The functions of these proteins were predicted using the protfun server. Our analysis showed that human proteins presented by MHC class II molecules are likely to be in the cell envelope, be a receptor or involved in immune responses. Presented proteins from bacteria and virus, on the other hand, are more likely to be involved in regulatory functions, translation, transcription as well as replication. These results can lead to better understanding the autoimmunity and the response to infections. PMID- 16212912 TI - Novel constructs of tuberculosis gene vaccine and its immune effect on mice. AB - A novel tuberculosis (TB) gene vaccine containing mouse granulocyte macrophage colony stimulating factor (mGM-CSF) and a TB antigen (Ag85A) was developed in this study. The genes encoding Ag85A and mGM-CSF were amplified by PCR respectively from the Ag85A-containing pBSby5 and pC-mGM-CSF. The genes were then cloned into two different polylinker sites of plasmid pIRES, forming a novel TB gene vaccine construct pI85AGM. Following transfection of pI85AGM plasmid into 7721 cell line by Lipofectamine(TM), the expression of Ag85A and GM-CSF proteins was identified by Western blotting or RT-PCR. Then Balb/c mice were inoculated with the recombinant pI85AGM, pI85A, pIGM or plasmid alone, respectively. The activities of CTL, NK cells and the Ag85A-stimulated proliferation of spleen cells were measured by MTT method. The serum antibody against Ag85A was detected by ELISA. The results showed that the Ag85A and GM-CSF proteins could be expressed in 7721 cell line and the activity of CTLs and the proliferation of spleen cells were significantly increased in the pI85AGM-immunized mice, indicating that the pI85AGM-immunized mice could generate specific immune responses to Ag85A. This study might provide possibility for developing novel anti-TB gene vaccine. PMID- 16212914 TI - Analysis tissue expression of IFN-gamma in IL-12 and/or IL-18 gene ablated naive mice. AB - Interleukin 12 (IL-12) and/or interleukin 18 (IL-18) gene ablated mice were applied for the investigation of the tissue expression of interferon gamma (IFN gamma). For IL-12(-/-), IL-18(-/-), IL-12(-/-)/18(-/-) and wt mice, reproductive performance were recorded and IFN-gamma concentrations in heart, lung, liver, spleen, kidney and serum were quantified by ELISA. There were no significant differences of IFN-gamma in heart, lung and kidney between 4 strains although control group was higher. It was observed that for IL-12(-/-) mice, compared with other 3 groups, IFN-gamma in liver and spleen were decreased (p < 0.05) and reproductive performance appeared to be impaired. Serum IFN-gamma level of IL-12( /-)/18(-/-) mice was significantly higher (p < 0.05). It was showed that IFN gamma productions under the normal condition were independent upon IL-12 and IL 18, its expressions in various tissues were different, and optimal IFN-gamma is necessary for the normal growth and development of mammals. This study is helpful for clinical cytokines therapy. PMID- 16212913 TI - Effects of activin A on the activities of the mouse peritoneal macrophages. AB - Activin A is a kind of pre-inflammatory factor that belongs to the transforming growth factor-beta (TGF-beta) superfamily. To investigate the effect and mechanism of activin A on the activities of mouse macrophages, the secretion of NO in the supernatant of cultured mouse peritoneal macrophages was examined by NO assay kit, and the expression of iNOS, ActRIIA and ARIP2 mRNA in mouse peritoneal macrophages was analyzed by RT-PCR. The results showed that activin A stimulated the secretion of NO and the expression of iNOS mRNA in non-activated mouse macrophages in a time- and dose-dependent manner. In contrast, activin A in the same concentration inhibited the secretion of NO in LPS-activated mouse macrophages in a dose-dependent manner. ActRIIA was highly expressed on macrophages, and activin A upregulated the ActRIIA mRNA expression in macrophages. Anti-ActRIIA antibody can block the secretion of NO from the macrophages stimulated by activin A. Furthermore, RT-PCR analysis revealed that activin A enhanced the ARIP2 mRNA expression in macrophages. These results indicated that Activin A may be a weak activator compared with LPS to mouse macrophages, and activin A may modulate the secretion of NO through ActRIIA-ARIP2 signal pathway in mouse macrophages. PMID- 16212915 TI - The late stage of T cell development within mouse thymus. AB - After positive selection and lineage commitment, the TCR alphabeta+ CD4/CD8 SP medullary thymocytes migrate into and reside in thymic medulla, where they undergo an ordered program of late stage of T cell functional maturation and negative selection to delete self-reactive clones by apoptosis. Accomplishment of this final differentiation pathway, a physiological T cell repertoire is formed: T cells acquire immunocompetence to respond to foreign antigens and tolerance to self-antigens, ready for the emigration to homing to the T cell regions of peripheral lymphoid organs and tissues. In this review, emphases are put on introducing the approaches applied in this area and our own observations. Basically, we have analyzed the late stage of medullary thymocyte phenotypic differentiation pathways of both CD4 SP and CD8 SP medullary thymocytes and the concomitant functional maturation pathway, in particular, of CD4 SP thymocytes. It is to provide a standard to compare the functional capacity of the cells at the developmental stages induced by different conditions. The cellular and molecular basis of this differentiation process has been partially described. PMID- 16212916 TI - Natural killer cells: biology and clinical use in cancer therapy. AB - Natural killer (NK) cells have the ability to mediate both bone marrow rejection and promote engraftment, as well as the ability to elicit potent anti-tumor effects. However the clinical results for these processes are still elusive. Greater understanding of NK cell biology, from activating and inhibitory receptor functions to the role of NK cells in allogeneic transplantation, needs to be appreciated in order to draw out the clinical potential of NK cells. Mechanisms of bone marrow cell (BMC) rejection are known to be dependent on inhibitory receptors specific for major histocompatibility complex (MHC) molecules and on activating receptors that have many potential ligands. The modulation of activating and inhibitory receptors may hold the key to clinical success involving NK cells. Pre-clinical studies in mice have shown that different combinations of activating and inhibitory receptors on NK cells can reduce graft versus-host disease (GVHD), promote engraftment, and provide superior graft versus-tumor (GVT) responses. Recent clinical data have shown that the use of KIR ligand incompatibility produces tremendous graft-versus-leukemia effect in patients with acute myeloid leukemia at high risk of relapse. This review will attempt to be a synthesis of current knowledge concerning NK cells, their involvement in BMT, and their use as an immunotherapy for cancer and other hematologic malignancies. PMID- 16212917 TI - MHC class I antigen presentation--recently trimmed and well presented. AB - Presentation of antigenic peptide to T cells by major histocompatibility complex (MHC) class I molecules is the key to the cellular immune response. Non-self intracellular proteins are processed into short peptides and transported into endoplasmic reticulum (ER) where they are assembled with class I molecules assisted by several chaperone proteins to form trimeric complex. MHC class I complex loaded with optimised peptides travels to the cell surface of antigen presentation cells to be recognised by T cells. The cells presenting non-self peptides are cleared by CD8 positive T cells. In order to ensure that T cells detect an infection or mutation within the target cells the process of peptide loading and class I expression must be carefully regulated. Many of the cellular components involved in antigen processing and class I presentation are known and their various functions are now becoming clearer. PMID- 16212918 TI - Immunotherapy with agonistic anti-CD137: two sides of a coin. AB - CD137 (4-1BB), a member of the TNF receptor superfamily, is an inducible T cell costimulatory receptor primarily expressed on activated CD4+ and CD8+ T cells. Agonistic monoclonal antibodies (mAbs) against CD137 greatly enhance T cell mediated immune responses against many types of tumors and viruses. Surprisingly, these agonists also showed therapeutic effects in several autoimmune diseases. These findings suggest that in different disease environments, CD137 engagement with agonist mAb in vivo can diametrically modulate immune response outcomes. Therefore, CD137 agonists represent a promising immunotherapeutic approach to a wide array of disparate immune disorders. However, CD137's potency in modulating immune response necessitates caution when targeting CD137 clinically. PMID- 16212919 TI - T lymphocyte co-signaling pathways of the B7-CD28 family. AB - The past years have witnessed significant advance in our understanding of critical roles of T cell co-signals in B7-CD28 family molecules in regulating T cell activation and tolerance. New co-signaling molecules have been identified and their functions have been delineated. These co-signaling pathways play overlapping and distinct regulatory roles at various stages of a T cell response. By expressing in appropriate time and location, these pathways have different regulatory functions through independent receptors or on different subsets of lymphocytes. Precise understanding of these pathways will allow the development of novel approaches to treatment of human diseases such as cancer, viral infection, autoimmune diseases and transplantation rejection. PMID- 16212920 TI - Hydrodynamic gene delivery of interleukin-22 protects the mouse liver from concanavalin A-, carbon tetrachloride-, and Fas ligand-induced injury via activation of STAT3. AB - Interleukin-22 (IL-22) is a recently identified T cell-derived cytokine whose biological significance remains obscure. Previously, we have shown that IL-22 plays a protective role in T cell-mediated hepatitis induced by Concanavalin A (Con A), acting as a survival factor for hepatocytes. In the present paper, we demonstrate that hydrodynamic gene delivery of IL-22 cDNA driven either by a liver-specific albumin promoter or a human cytomegalovirus (CMV) promoter results in IL-22 protein expression, STAT3 activation, and expression of several anti apoptotic proteins, including Bcl-xL, Bcl-2, and Mcl-1 in the liver. Immunohistochemical analysis reveals that IL-22 protein expression is mainly detected in the cytoplasm of hepatocytes. Overexpression of IL-22 by hydrodynamic gene delivery significantly protects against liver injury, necrosis, and apoptosis induced by administration of Con A, carbon tetrachloride (CCl4), or the Fas agonist Jo-2 mAb. Western blot analyses show that overexpression of IL-22 significantly enhances activation of STAT3 and expression of Bcl-xL, Bcl-2, and Mcl-1 proteins in liver injury induced by Con A. In conclusion, hydrodynamic gene delivery of IL-22 protects against liver injury induced by a variety of toxins, suggesting the therapeutic potential of IL-22 in treating human liver disease. PMID- 16212922 TI - Kinetics of the phenotype and function of murine peritoneal macrophages following acute inflammation. AB - This study was undertaken to have a better understanding for the process and the underlying mechanisms to limit macrophage activation and population of activated macrophages. A comprehensive kinetics of cytokine production was performed in murine peritoneal macrophages recovered from Balb/c mice at various time during the course of an intraperitoneal injection with thioglycollate (TG). The expression of cell surface molecules such as MHC-I, MHC-II, B7-1 and B7-2 of these macrophages were also determined by flow cytometry. The present findings of our research suggested that the population of activated macrophages and the activation of macrophages (including cytokines production and expression of cell surface functional molecules) were strictly controlled during inflammation process. This is one of the important mechanisms to retain the host homeostasis. PMID- 16212921 TI - The grape component resveratrol interferes with the function of chemoattractant receptors on phagocytic leukocytes. AB - Resveratrol (3,5,4'-trihydroxystilbene) (RV) is a constituent of grape seeds with anti-inflammatory and anti-oxidant activities. In this study, we examined the capacity of RV to modulate the function of G protein-coupled chemoattractant receptors, which play important roles in inflammation and immune responses. RV, over a non-cytotoxic concentration range, inhibited chemotactic and calcium mobilization responses of phagocytic cells to selected chemoattractants. At low micromolar concentrations, RV potently reduced superoxide anion production by phagocytic leukocytes in response to the bacterial chemotactic peptide fMLF, a high affinity ligand for formylpeptide receptor FPR, and A beta42, an Alzheimer's disease-associated peptide and a ligand for the FPR variant FPRL1. In addition, RV reduced phosphorylation of extracellular signal-regulated kinase (ERK1/2) and the activation of nuclear factor NF-kappaB induced by formylpeptide receptor agonists. These results suggest that the inhibition of the function of chemoattractant receptors may contribute to the anti-inflammatory properties of RV. Thus, RV may be therapeutically promising for diseases in which activation of formylpeptide receptors contributes to the pathogenic processes. PMID- 16212923 TI - Type two cytokines predominance of human lung cancer and its reverse by traditional Chinese medicine TTMP. AB - Type 2 cytokines are usually predominant in tumor patients and associated with tumor progression. To explore whether reversing of type 2 predominance could be a promising strategy in tumor immunotherapy, PBMC of 35 lung cancer patients and 19 healthy subjects were prepared and subjected to be examined for cytokine secretion and gene expression. Tetra-Methylpyrazine (TTMP), extracted from a traditional Chinese medicinal herb which has been used in clinic to reverse the Th2 status of cancer patients in China, was added to PBMC culture. Determined by RT-PCR, the positive percentages of mRNA expression of type 1 cytokines (8.6% for IFN-gamma and 11.4% for IL-2) were lower than those of type 2 cytokines (71.4% for IL-4, 60% for IL-6 and 80% for IL-10) in patients' PBMCs. The potential of gene expressing (measured as relative intensity to the ratio of beta-actin) in the patients for type 1 cytokines was also in a low level (0.111 for IFN-gamma, 0.119 for IL-2) in comparison with a relative high level for type 2 cytokines (0.319 for IL-4, 0.303 for IL-6 and 0.377 for IL-10). Meanwhile, both positive percentage and relative intensity of gene expression were lower for a type 1 cytokine-related transcription factor T-bet (31.4% and 0.142, respectively) than those for type 2 cytokine-related GATA3 (85.7% and 0.378, respectively). The blood serum levels of IFN-gamma and IL-2 in the patients were slightly lower but not significantly when compared with healthy control. In contrast, the levels IL 4 and IL-6 in patients were significantly higher than those in healthy subjects by ELISA analysis. TTMP could enhance supernatant concentration and gene expression levels of IFN-gamma, IL-2 and T-bet, but reduce those of type 2 cytokines. These results demonstrate that the lung cancer patients had a predominant expression of type 2 cytokines and TTMP could reverse the type 2 dominant status, which might offer an alternative therapeutic regime for lung cancer patients. PMID- 16212925 TI - New fluorescence markers to distinguish co-infecting Trypanosoma brucei strains in experimental multiple infections. AB - Multiple-genotype infections are increasingly recognized as important factors in disease evolution, parasite transmission dynamics, and the evolution of drug resistance. However, the distinction of co-infecting parasite genotypes and the tracking of their dynamics have been difficult with traditional methods based on various genotyping techniques, leaving most questions unaddressed. Here we report new fluorescence markers of various colours that are inserted into the genome of Trypanosoma brucei to phenotypically label live parasites of all life cycle stages. If different parasite strains are labelled with different colours they can be easily distinguished from each other in experimental studies. A total of 10 T. brucei strains were successfully transfected with different fluorescence markers and were monitored in culture, tsetse flies and mice, to demonstrate stability of marker expression. The use of fluorescence activated cell sorting (FACS) allowed rapid and accurate identification of parasite strains labelled with different markers. Cell counts by FACS were virtually identical to counts by traditional microscopy (n=75, Spearman's rho: 0.91, p<0.0001) but were considerably faster and had a significantly lower sampling error (66% lower, d.f.=73, t=-17.1, p<0.0001). Co-infecting strains transfected with fluorescence genes of different colour were easily distinguished by eye and their relative and absolute densities were reliably counted by FACS in experimental multiple infections in mice. Since the FACS can simultaneously determine the population sizes of differently labelled T. brucei strains or subspecies it allows detailed and efficient tracking of multiple-genotype infections within a single host or vector individual, enabling more powerful studies on parasite dynamics. In addition, it also provides a simple way to separate genotypes after experimental mixed infections, to measure responses of the single strains to an applied treatment, thus eliminating the need for laborious cloning steps. The markers presented broaden the spectrum of tools available for experimental studies on multiple-genotype infections. They are fundamentally different from isoenzyme analysis and other genotyping approaches in that they allow the distinction of parasite genotypes based on an easily recognizable phenotypic trait. They will be of specific interest to researches addressing ecological, evolutionary and epidemiological questions using trypanosomes as an experimental system. PMID- 16212924 TI - Anti-CD137 monoclonal antibody promotes the direct anti-tumor effect mediated by peripheral blood-derived human dendritic cells in vitro. AB - CD137, a costimulatory factor of TNFR family, is expressed on activated T cells and freshly isolated mouse dendritic cells (DCs). To date, there are only limited data dealing with the expression and the effect of CD137 on human DCs. We report in this work that CD137 can coexpress with CD137L on immature peripheral blood derived human DCs (9.77%). The mature DCs stimulated by LPS showed a much higher level of CD137 expression (36.06%). Ligation of CD137 on the surface of DCs with anti-CD137 monoclonal antibody (mAb) could enhance the direct anti-tumor effect mediated by human DCs in vitro. The agonistic anti-CD137 mAb was able to elevate by about 20% of the DC-mediated inhibition of tumor growth in five tumor cell lines. These results indicate that the appliance of anti-CD137 mAb might be used as a new strategy for tumor immunotherapy. PMID- 16212926 TI - Combined methods for the study of water contact behavior in a rural schistosomiasis-endemic area in Brazil. AB - A new combined methodology consisting of direct observation and two types of interviews (internal and external interviews) was evaluated for use in exposure risk assessment in schistosomiasis. Specific objectives were to determine its usefulness in achieving equitable coverage of gendered exposure risk and its efficiency in identifying water contact behavior in a rural area in Brazil with different settlement patterns, land use and domestic water supplies. Of the 2476 water contacts recorded, 1223 (49.4%) were identified by direct observation, 946 (38.2%) by internal interviews and 307 (12.4%) by external interviews. Significantly longer mean durations of contacts were recorded for females and greater mean percentage of body surface exposed for males (P<0.01), reflecting differences in gendered water contact activities. Direct observation identified slightly more male contacts, external interviews significantly more male contacts (P<0.006), and internal interviews moderately more female contacts. The three methods recorded mean numbers of contacts and mean TBM (total body minutes) per person, declining with age. Significant differences were found between the three methods in regard to frequency and/or intensity of washing clothes, fetching water, washing utensils, washing multiple parts of the body, and bathing. The three methods also recorded differentially frequencies and exposure intensities in the three study communities, among different age groups, by gender and for individual study members. These activity-, locality-, age/gender- and person specific patterns reflect the relative efficiency and complementarity of the three methods in settlements with different land use, access to streams and water supplies. PMID- 16212927 TI - Resonance Rayleigh scattering method for the determination of tetracycline antibiotics with uranyl acetate and water blue. PMID- 16212928 TI - A series of bidirectional tetracycline-inducible promoters provides coordinated protein expression. AB - Coordinated control of protein expression is highly desirable for functional genomics. Here we show a widely applicable approach to construction and use of custom bidirectional promoters capable of reproducible coexpression of two proteins. The use of a bidirectional promoter system overcomes many of the limitations of current coexpression systems such as unpredictable upstream and downstream expression ratios mediated by an internal ribosome entry site. We present examples of tetracycline-inducible, bidirectional promoter systems that produce simultaneous and rapid coinduction of two separate reporters to predictable levels and ratios. Steric blocking of transcription initiation by simple tetracycline repressors, rather than the use of transcriptional activator/repressor domain fusions, makes the system described here superior for investigating downstream transcriptional consequences of protein expression. PMID- 16212929 TI - Whole genome amplification strategy for forensic genetic analysis using single or few cell equivalents of genomic DNA. AB - Evidentiary items sometimes contain an insufficient quantity of DNA for routine forensic genetic analysis. These so-called low copy number DNA samples (< 100 pg of genomic DNA) often fall below the sensitivity limitations of routine DNA analysis methods. Theoretically, one way of making such intractable samples amenable to analysis would be to increase the number of starting genomes available for subsequent STR (short tandem repeat) analysis by a whole genome amplification strategy (WGA). Although numerous studies employing WGA have focused primarily on clinical applications, few in-depth studies have been conducted to evaluate the potential usefulness of these methods in forensic casework. After an initial evaluation of existing methods, a modified WGA strategy was developed that appears to have utility for low copy number forensic casework specimens. The method employs a slight, but important, modification of the "improved primer extension preamplification PCR" method (I-PEP-PCR), which we term mIPEP (modified-I-PEP-PCR). Complete autosomal STR and Y-STR (Y chromosome short tandem repeat) profiles were routinely obtained with 5 pg of template DNA, which is equivalent to 1-2 diploid cells. Remarkably, partial Y- and autosomal STR profiles were obtained from mIPEP-treated DNA recovered from bloodstains exposed to the outside environment for 1 year whereas non-mIPEP-treated samples did not produce profiles. STR profiles were obtained from contact DNA from single dermal ridge fingerprints when the DNA was subjected to prior mIPEP amplification but not when the mIPEP step was omitted. PMID- 16212930 TI - ADP-ribosyl cyclase and GDP-ribosyl cyclase activities are not always equivalent: impact on the study of the physiological role of cyclic ADP-ribose. PMID- 16212931 TI - Injuries from slips and trips in construction. AB - Construction injuries preceded by a slip or trip were documented using data from the building of the Denver International Airport (Denver, Colorado, USA), the largest construction project in the world at the time. Slips and trips occurred at a rate of 5/200,000 h worked accounting for 18% of all injuries and 25% of workers' compensation payments, or more than $10 million. Slips contributed to the vast majority (85%) of same-level falls and over 30% of falls from height, as well as a significant number of musculoskeletal injures sustained after slipping or tripping but without falling. The injury burden would have been under recognized in analyses of most coded compensation records. In contrast to other types of injuries, the most common contributing factors were environmental in nature including conditions of walking and working surfaces, terrain and weather. Due to the very dynamic nature of construction work, reducing slips and trips will require a focus on environmental and organizational solutions that evolve as the site changes and the construction project evolves. PMID- 16212932 TI - The PufX protein of Rhodobacter capsulatus affects the properties of bacteriochlorophyll a and carotenoid pigments of light-harvesting complex 1. AB - A pufX gene deletion in the purple bacterium Rhodobacter capsulatus causes a severe photosynthetic defect and increases core light-harvesting complex (LH1) protein and bacteriochlorophyll a (BChl) levels. It was suggested that PufX interrupts the LH1 alpha/beta ring around the reaction centre, allowing quinone/quinol exchange. However, naturally PufX(-) purple bacteria grow photosynthetically with an uninterrupted LH1. We discovered that substitutions of the Rhodobacter-specific LH1 alpha seryl-2 decrease carotenoid levels in PufX( )R. capsulatus. An LH1 alphaS2F mutation improved the photosynthetic growth of a PufX(-) strain lacking the peripheral LH2 antenna, although LH1 BChl absorption remained above wild-type, suggesting that Rhodobacter-specific carotenoid binding is involved in the PufX(-) photosynthetic defect and LH1 expansion is not. Furthermore, PufX overexpression increased LH1-like BChl absorption without inhibiting photosynthetic growth. PufX(+) LH1 alphaS2-substituted mutant strains had wild-type carotenoid levels, indicating that PufX modulates LH1 carotenoid binding, inducing a conformational change that favours quinone/quinol exchange. PMID- 16212933 TI - Fast, efficient reconstitution of the cyclooxygenases into proteoliposomes. AB - To study the physical and catalytic properties of purified membrane proteins, it is often necessary to reconstitute them into lipid bilayers. Here, we describe a fast efficient method for the direct incorporation of cyclooxygenase-1 and -2 (COX-1 and -2) isozymes into liposomes without loss of activity. Purified COX-1 and -2 spontaneously incorporate into large unilamellar vesicles produced from a mixture of DOPC:DOPS (7:3) that has been doped with oleic acid. When incorporation was measured by comparing cyclooxygenase activity to total phospholipid in the proteoliposomes, molar reconstitution ratios of 1000:1 (phospholipid:COX) were obtained. Electron paramagnetic resonance spectroscopic spin counting analysis of proteoliposomes formed with nitroxide spin-labeled COX 2 gave a nearly identical phospholipid:COX ratio, confirming that incorporation had no effect on enzyme activity, and demonstrating that the efficiency of protein incorporation is sufficient for EPR spectroscopic analysis. The spontaneous incorporation of cyclooxygenase into intact liposomes allows only insertion into the outer leaflet for this monotopic enzyme, an orientation confirmed by immunogold staining of the proteoliposomes. This method of reconstitution into liposomes may be generally applicable to the class of monotopic integral membrane proteins typified by the cyclooxygenase isozymes. PMID- 16212934 TI - Metacognitions in patients with hallucinations and obsessive-compulsive disorder: the superstition factor. AB - On the basis of the analogy between intrusive thoughts and auditory hallucinations established by Morrison et al. [(1995). Intrusive thoughts and auditory hallucinations: a cognitive approach. Behavioural and Cognitive Psychotherapy, 23, 265-280], the present work compares the metacognitive beliefs and processes of five groups of patients (current hallucinators, never hallucinated people with a diagnosis of schizophrenia, recovered hallucinators, obsessive-compulsive disorder (OCD) patients, and a clinical control group) and a non-clinical group. The results show that of the five metacognitive factors considered in this study, two were found to be different in the current hallucinators group in comparison to any other group in the design. Likewise, it is found that the metacognitive beliefs of the current hallucinators coincide with those of the OCD patients in various factors, particularly that relating to superstition, and this is interpreted as lending support to the model of Morrison et al. (1995). Furthermore, the results are discussed in the light of existing research on Thought-Action Fusion, stressing the role that may be played by superstitious beliefs and magical thinking in auditory hallucinations and OCD. PMID- 16212935 TI - Candida albicans protein analysis during hyphal differentiation using an integrative HA-tagging method. AB - In order to understand the biological complexity inherent to the pathogenicity of Candida albicans, gene expressions need to be analyzed at the protein level. We employed an epitope-tagging technique in a set of C. albicans ORF, and constructed fifteen strains which expressed HA-tagged proteins. These efforts permitted us to identify differentially synthesized proteins during the hyphal differentiations. ICL1, MLS1, and WAP1, all of which are known to be hypha induced at the transcript level, were indeed found to be up-regulated at the protein level. We also identified CaeIF4G, CaTPO5, and CaZRT1, the protein levels of which were increased during hyphal transition, and CaERB1, the protein level of which was reduced consistently. The hypha-induced protein level of CaeIF4G was closely associated with the cellular hyphal phenotype. CaeIF4G overexpression was shown to result in hyperfilamentation in C. albicans. CaeIF4E, which was constitutively expressed during the hyphal development, exhibited no overexpression phenotype. HA-tagged strains were also utilized in our analysis of C. albicans proteins in a co-culture of macrophage and C. albicans. Five genes were found to be expressed differentially during the macrophage co-cultures. Our approaches proved to be rather useful under yeast culture conditions as well as in co-cultures of macrophage and C. albicans. PMID- 16212936 TI - The whcE gene of Corynebacterium glutamicum is important for survival following heat and oxidative stress. AB - In this study, we have analyzed an ORF from Corynebacterium glutamicum, which codes for a homologue of the Streptomyces coelicolor WhiB-family of proteins known to be involved in sporulation. This ORF encoded a putative protein which harbors a helix-turn-helix DNA-binding motif and a probable redox-sensing motif, and has been designated whcE. We constructed a whcE mutant strain and analyzed the strain under a variety of growth conditions. This mutant strain exhibited a prolonged lag phase and earlier death within the stationary phase, suggesting that the relevant gene may play a role in both growth adaptation and stress responses. Further analysis determined that the mutant strain was not only sensitive with regard to survival under heat stress, but was also markedly susceptible to thiol-specific oxidant diamide and redox cycling compounds, including menadione and plumbagin. The mutant strain also exhibited reductions in thioredoxin reductase activity, which indicates that the trxB gene encoding thioredoxin reductase is under the control of WhcE. Expression of whcE was stimulated during the stationary phase of cell growth and could be modulated by diamide. We also delineated the relationship between whcE and the sigH gene, which is located downstream of whcE, and has been shown to be involved in heat stress responses, via the encoding of an ECF sigma factor. In a sigH mutant strain, the whcE gene was no longer expressed, thereby suggesting that the sigmaH sigma factor is involved in whcE expression. Our results suggest that WhcE functions as a transcription factor which can activate the trxB gene, as well as other genes, possibly by sensing redox changes during the metabolic downshifting of cells from exponential growth to the stationary phase, whereas sigmaH appears to function as the sigma factor for these genes, including whcE. PMID- 16212937 TI - hCOX18 and hCOX19: two human genes involved in cytochrome c oxidase assembly. AB - We identified the human homologues of yCOX18 and yCOX19, two Saccharomyces cerevisiae genes involved in the biogenesis of mitochondrial respiratory chain complexes. In yeast, these two genes are required for the expression of cytochrome c oxidase: Cox18p catalyses the insertion of Cox2p COOH-tail into the mitochondrial inner membrane, and Cox19p is probably involved in metal transport to the intermembrane space. Both hCox18p and hCox19p present significant amino acid identity with the corresponding yeast polypeptides and reveal highly conserved functional domains. In addition, their subcellular localization is analogous to that of the yeast proteins. These data strongly suggest that the human gene products share similar functions with their yeast homologues. These two COX-assembly genes represent new candidates for mutational analysis in patients with isolated COX deficiency of unknown etiology. PMID- 16212938 TI - Caffeine-mediated enhancement of glucocorticoid receptor activity in human osteoblastic cells. AB - Caffeine-containing beverage consumption has been reported to be associated with an increased risk for osteoporosis. Since the glucocorticoid receptor (GR) is a major factor in the induction of osteoporosis we analyzed whether caffeine may act via altering GR function. Applying a reporter gene assay we provide evidence that caffeine drastically amplifies GR transcriptional activity in human osteoblastic cells. Substances that increase the intracellular cyclic AMP concentration also strengthen the transactivity of the GR and coincubation with caffeine further reinforces this potentiation, indicating that caffeine-mediated enhancement of GR transcriptional function is due to the inhibitory activity of caffeine on the cyclic AMP phosphodiesterase. Our data suggest evidence for a hitherto unrecognized crosstalk between caffeine-modulated signalling and GR initiated gene expression in human osteoblastic cells and could provide the molecular basis for the role of caffeine in osteoporosis. PMID- 16212939 TI - N-Glycosylation of secretion enhancer peptide as influencing factor for the secretion of target proteins from Saccharomyces cerevisiae. AB - hIL-1beta-derived polypeptide, when fused to the N-terminal end of target proteins, exerts a potent secretion enhancer function in Saccharomyces cerevisiae. We investigated the effect of N-glycosylation of the secretion enhancer peptide on the secretion of target proteins. The N-terminal 24 amino acids (Ser5-Ala28) of human interleukin 1beta (hIL-1beta) and interleukin 1 receptor antagonist (IL-1ra) were used as secretion enhancer for synthesizing recombinant human granulocyte-colony stimulating factor (rhG-CSF) from S. cerevisiae. The mutation of potential N-glycosylation site, by substituting Gln for either Asn7 of N-terminal 24 amino acids of hIL-1beta (Asn7Gln) or Asn84 of IL-1ra (Asn84Gln), resulted in a dramatic reduction of rhG-CSF secretion efficiency. In contrast, the mutant containing an additional N-glycosylation site on the N-terminal 24 amino acids of hIL-1beta (Gln15Asn) secreted twice as much rhG-CSF into culture media as wild type hIL-1beta. These results show that N glycosylation of the secretion enhancer peptide plays an important role in increasing the secretion efficiency of the downstream target proteins. The results also suggest that judicious choice of enhancer peptide and the control of its glycosylation could be of general utility for secretory production of heterologous proteins from S. cerevisiae. PMID- 16212941 TI - Regulation of tissue transglutaminase by prolonged increase of intracellular Ca2+, but not by initial peak of transient Ca2+ increase. AB - Tissue transglutaminase (tTGase) is a member of calcium-dependent transamidation enzyme family, but a detailed regulation mechanism of tTGase by intracellular Ca(2+) is not clearly understood. Arachidonic acid (AA) and maitotoxin (MTX) activated tTGase in a dose- and time-dependent manner. Transfection of tTGase siRNA largely inhibited tTGase expression and tTGase activation by MTX. AA induced an initial increase of intracellular Ca(2+) followed by a prolonged increase. Removal of extracellular Ca(2+) with EGTA blocked the prolonged Ca(2+) increase in response to AA, although the initial Ca(2+) increase remained. In contrast, EGTA completely blocked the increase of intracellular Ca(2+) by MTX. The activation of tTGase by AA or MTX was significantly inhibited by EGTA. Moreover, EGTA prevented the prolonged increase of intracellular Ca(2+) and tTGase activation by lysophosphatidic acid, but had no effect on the initial Ca(2+) increase. These results suggested that tTGase is regulated by the prolonged increase of intracellular Ca(2+) originated from Ca(2+) influx, rather than by the initial peak of transient Ca(2+) increase. PMID- 16212940 TI - Biochemical characterization of recombinant phosphoglucose isomerase of Mycobacterium tuberculosis. AB - Phosphoglucose isomerase (PGI) is a well-characterized ubiquitous enzyme involved in the glycolytic pathway. It catalyzes the reversible isomerization of D glucopyranose-6-phosphate and D-fructofuranose-6-phosphate and is present in all living cells. However, there is interspecies variation at the level of the primary structure which sometimes produces heterogeneity at the structural and functional levels. In order to evaluate and characterize the mycobacterial PGI, the gene encoding the PGI from Mycobacterium tuberculosis H37Rv was cloned in pET 22b(+) vector and expressed in Escherichia coli. The target DNA was PCR amplified from the bacterial artificial chromosome using specific primers and cloned under the control of T7 promoter. Upon induction with IPTG, the recombinant PGI (rPGI) expressed partly as soluble protein and partly as inclusion bodies. The rPGI from the soluble fraction was purified to near homogeneity by ion-exchange chromatography. Mass spectrum analysis of the purified rPGI revealed its mass to be 61.45 kDa. The purified rPGI was enzymatically active and the specific activity was 600 U/mg protein. The K(m) of rPGI was determined to be 0.318 mM for fructose-6-phosphate and the K(i) was 0.8 mM for 6-phosphogluconate. The rPGI exhibited optimal activity at 37 degrees C and pH 9.0, and did not require mono- or divalent cations for its activity. PMID- 16212942 TI - In vivo functional characterization of the SARS-Coronavirus 3a protein in Drosophila. AB - The Severe Acute Respiratory Syndrome-Coronavirus (SARS-CoV) 3a locus encodes a 274 a.a. novel protein, and its expression has been confirmed in SARS patients. To study functional roles of 3a, we established a transgenic fly model for the SARS-CoV 3a gene. Misexpression of 3a in Drosophila caused a dominant rough eye phenotype. Using a specific monoclonal antibody, we demonstrated that the 3a protein displayed a punctate cytoplasmic localization in Drosophila as in SARS CoV-infected cells. We provide genetic evidence to support that 3a is functionally related to clathrin-mediated endocytosis. We further found that 3a misexpression induces apoptosis, which could be modulated by cellular cytochrome c levels and caspase activity. From a forward genetic screen, 78 dominant 3a modifying loci were recovered and the identity of these modifiers revealed that the severity of the 3a-induced rough eye phenotype depends on multiple cellular processes including gene transcriptional regulation. PMID- 16212943 TI - Involvement of 5-HT1B receptors within the ventral tegmental area in ethanol induced increases in mesolimbic dopaminergic transmission. AB - Evidence suggests that 5-hydroxytriptamine-1B (5-HT1B) receptors play a role in modifying ethanol's reinforcing effects and voluntary intake, and that 5-HT1B receptors within the ventral tegmental area (VTA) are involved in regulation of mesolimbic dopaminergic neuronal activity. Since increased mesolimbic dopaminergic transmission has been implicated in ethanol's reinforcing properties, this study was designed to assess the involvement of VTA 5-HT1B receptors in mediating the stimulatory effects of ethanol on VTA dopaminergic neurons. Dual-probe microdialysis was performed in freely moving adult Sprague Dawley rats with one probe within the VTA and the other within the ipsilateral nucleus accumbens (NACC). Dopamine (DA) levels in dialysates from both areas, as the index of the activity of mesolimbic DA neurons, were measured simultaneously. The results showed that intraperitoneal injection of ethanol at the doses of 1 and 2 g/kg increased extracellular DA concentrations in both the VTA and the NACC, suggesting increased DA neuronal activity. These ethanol-induced increases of the DA release in the VTA and the NACC were significantly attenuated by intra tegmental infusion of SB 216641 (a 5-HT(1B) receptor antagonist), but not BRL 15572 (a 5-HT(1D/1A) receptor antagonist) or WAY 100635 (a 5-HT1A receptor antagonist). Administration of ethanol at the same doses did not significantly alter extracellular levels of GABA in the VTA. The results also showed that intra tegmental infusion of CP 94253, a 5-HT1B receptor agonist, significantly prolonged the effects of ethanol on NACC DA. The results suggest that blockade and activation of VTA 5-HT1B receptors attenuates and potentiates the neurochemical effects of ethanol, respectively, and support the suggestion that VTA 5-HT(1B) receptors may be involved in part in mediating the activating effects of ethanol on mesolimbic DA neurons. PMID- 16212944 TI - 17beta-Estradiol attenuates blood-brain barrier disruption induced by cerebral ischemia-reperfusion injury in female rats. AB - Disruption of blood-brain barrier (BBB), mediated through matrix metalloproteinases (MMPs), is a critical event during cerebral ischemia. While neuroprotective effects of estrogens have been well established in ischemic stroke models, the effects of estrogens on BBB integrity remain to be elucidated. In the present study, we determined effects of 17beta-estradiol (E2) on BBB disruption induced by transient focal cerebral ischemia and its effects on MMP2 and MMP9 activation. Transient cerebral ischemia was induced by middle cerebral artery (MCA) occlusion for 1 h followed by reperfusion in ovariectomized rats. E2 (100 microg/kg) or vehicle was administered 2 h before MCA occlusion. BBB integrity was determined by fluorescent detection of extravasated Evans blue. In separate experiments, effect of E2 on MMP2 and MMP9 expression and activation was determined by immunoblot and MMPs activity assay. E2 treatment prevented more than 50% and 30% of BBB disruption in the ischemic cortex and subcortex at 4 h after reperfusion, respectively. MMP2 and MMP9 expression was elevated at 2 h and peaked at 4 h after reperfusion in the ischemic cortex, which was markedly reduced by E2 treatment. E2 treatment also attenuated the increase of MMPs activity induced by ischemia-reperfusion injury. In conclusion, estrogens could attenuate BBB disruption induced by transient cerebral ischemia, by inhibition of MMP2 and MMP9 activation. Our results suggest an important role of estrogens as multiple targeting protectants against ischemic stroke on cellular as well as vascular components of central nervous system. PMID- 16212945 TI - Rivastigmine is a potent inhibitor of acetyl- and butyrylcholinesterase in Alzheimer's plaques and tangles. AB - Acetylcholinesterase and butyrylcholinesterase activities emerge in association with plaques and tangles in Alzheimer's disease. These pathological cholinesterases, with altered properties, are suggested to participate in formation of plaques. The present experiment assessed the ability of rivastigmine, a clinically utilized agent that inhibits acetylcholinesterase and butyrylcholinesterase activities, to inhibit cholinesterases in plaques and tangles. Cortical sections from cases of Alzheimer's disease were processed using cholinesterase histochemistry in the presence or absence of rivastigmine. Optical densities of stained sections were utilized as a measure of inhibition. The potency of rivastigmine was compared with those of other specific inhibitors. Optimum staining for cholinesterases in neurons and axons was obtained at pH 8.0. Cholinesterases in plaques, tangles and glia were stained best at pH 6.8. Butyrylcholinesterase-positive plaques were more numerous than acetylcholinesterase-positive plaques. Rivastigmine inhibited acetylcholinesterase in all positive structures in a dose-dependent manner (10( 6)-10(-4) M). However, even at the highest concentration, faint activity remained. In contrast, rivastigmine resulted in complete inhibition of butyrylcholinesterase in all structures at 10(-5) M. Rivastigmine was equipotent to the specific acetylcholinesterase inhibitor BW284C51 and more potent than the butyrylcholinesterase inhibitors iso-OMPA and ethopropazine. In conclusion, rivastigmine is a potent inhibitor of acetylcholinesterase and a more potent inhibitor of butyrylcholinesterase in plaques and tangles. Unlike other cholinesterase inhibitors tested, rivastigmine inhibited cholinesterases in normal and pathological structures with the same potency. Thus, at the therapeutic concentrations used, rivastigmine is likely to result in inhibition of pathological cholinesterases, with the potential of interfering with the disease process. PMID- 16212946 TI - Chronic morphine exposure affects the visual response properties of V1 neurons in cat. AB - Chronic opiate exposure leads to maladaptive changes in brain function. In view of the localization of opiate receptors in mammalian visual system, chronic opiate exposure is likely to affect the visual responses properties of V1 neurons. Using in vivo single-unit recording, we here showed that chronic morphine treatment resulted in the functional abnormality of primary visual cortical cells. When compared with saline-treated (as control) cats, cortical neurons in morphine-treated cats exhibited higher spontaneous activity, lower signal-to-noise ratios and weaker orientation and direction selectivity. However, re-exposure with morphine could significantly improve the function of V1 neurons in morphine-treated cats. These findings demonstrated that chronic morphine treatment could significantly degrade the response properties of V1 neurons and may lead to a function dependence on morphine in visual cortical cells. PMID- 16212947 TI - Neuroanatomical distribution and colocalisation of nuclear receptor corepressor (N-CoR) and silencing mediator of retinoid and thyroid receptors (SMRT) in rat brain. AB - The two structurally related nuclear receptor corepressor (N-CoR) and silencing mediator of retinoid and thyroid receptors (SMRT) proteins have been found to differentially affect the transcriptional activity of numerous nuclear receptors, such as thyroid hormone, retinoic acid and steroid receptors. Because of the numerous effects mediated by nuclear receptors in brain, it is of interest to extend these in vitro data and to explore the cellular distribution of both corepressors in brain tissue. We therefore examined, using in situ hybridisation, whether the relative abundance of these two functionally distinct corepressors differed in rat brain and pituitary. We find that although both N-CoR and SMRT transcripts are ubiquitously expressed in brain, striking differences in their respective levels of expression could be observed in discrete areas of brain stem, thalamus, hypothalamus and hippocampus. Using dual-label immunofluorescence, we examined in selected glucocorticoid sensitive areas involved in the regulation of the hypothalamus-pituitary-adrenal axis activity, the respective protein abundance of N-CoR and SMRT. Protein abundance was largely concurrent with the mRNA expression levels, with SMRT relatively more abundant in hypothalamus and brain stem areas. Colocalisation of N-CoR and SMRT was demonstrated by confocal microscopy in most areas studied. Taken together, these findings are consistent with the idea that the uneven neuroanatomical distribution of N-CoR and SMRT protein may contribute to the site-specific effects exerted by hormones, such as glucocorticoids, in the brain. PMID- 16212949 TI - Dynamic site heterogeneity in amorphous maltose and maltitol from spectral heterogeneity in erythrosin B phosphorescence. AB - We have used phosphorescence from erythrosin B (tetraiodofluorescein) dispersed in thin films of either maltose or maltitol to investigate the physical properties of these amorphous pure sugar matrixes. Intensity decays collected as a function of emission wavelength over the range from 640 to 720 nm were analyzed using a stretched exponential kinetic model in which the lifetime (tau) and the stretching exponent (beta) were the physically relevant parameters. The lifetimes varied systematically with emission wavelength in both matrixes. Analysis of the temperature dependence of the lifetime at each wavelength provided an estimate of the activation energy for nonradiative quenching of the triplet state; the activation energy also varied with emission wavelength. In addition, time resolved emission spectra exhibited a blue shift with time following excitation. These data support a photophysical model in which probes are distributed among sites that vary in terms of overall molecular mobility and in which sites with lower rates of dipolar relaxation also have lower rates of collisional quenching of the erythrosin triplet state. The amorphous matrix of both maltose and maltitol in both the glass and the melt state is thus characterized by dynamic site heterogeneity in which different sites vary in terms of their overall molecular mobility. PMID- 16212948 TI - Glycyl-glutamine in nucleus accumbens reduces ethanol intake in alcohol preferring (P) rats. AB - Opioid peptides and glycyl-glutamine (Gly-Gln) have been implicated in the control of ethanol consumption. A recognized beta-endorphin cleavage product, Gly Gln, inhibits voluntary alcohol consumption when microinjected into the nucleus accumbens (AcbSh) of P rats. To evaluate the site-specific efficacy of Gly-Gln on ethanol consumption following AcbSh application, ethanol preferring (P) rats were allowed to establish individual baseline ethanol/water consumption utilizing a voluntary self-administration paradigm. Subsequent to baseline ethanol consumption being established, bilateral guide cannulae were stereotaxically implanted +1 mm dorsal to the AcbSh for subsequent Gly-Gln (100 nmol/microl) or saline vehicle (1 microl) injections. Alcohol intake, body weight, and water intake were measured at 24 h post-injection intervals. Unilateral Gly-Gln injections reduced ethanol consumption 35.6% (P < 0.05) from pre-established baseline consumption (6.24 +/- 0.64 g/kg to 4.06 +/- 0.28 g/kg). Bilateral Gly Gln injections further reduced consumption to 51.9% (6.4 +/- 1.0 g/kg to 3.08 +/- 0.65 g/kg at 24 h (P < 0.01) below established baseline values within 24 h without significant changes in body weight or water consumption. Also, the amino acid constituents of the dipeptide had no influence on ethanol consumption behavior; however, Gly-Gln efficacy was shown to be comparable to central beta endorphin-(1-27) or intraperitoneal (i.p.) naltrexone-induced suppression of ethanol intake. These data indicate that the AcbSh exhibits a site-specific sensitivity to the suppressive actions of Gly-Gln or beta-endorphin-(1-27) injections that modulate voluntary ethanol consumption in P rats. These findings support the broader concept that select forebrain opioid-responsive neural sites may influence the development or expression of alcohol abuse syndromes in animal models or humans. PMID- 16212950 TI - Glycosyl trichloroacetylcarbamate: a new glycosyl donor for O-glycosylation. AB - Glycosyl trichloroacetylcarbamates, readily obtained by reacting 1-hydroxy sugars with trichloroacetylisocyanate, have been found as excellent glycosyl donors, and the corresponding O-glycosides are formed in good to excellent yields with a fairly good degree of selectivity. PMID- 16212951 TI - Biphasic effect of p21Cip1 on smooth muscle cell proliferation: role of PI 3 kinase and Skp2-mediated degradation. AB - OBJECTIVE: Proliferation of vascular smooth muscle cells (VSMC) is an important event in atherogenesis, in-stent restenosis and late vein-graft failure. Cell cycle progression is positively regulated by cyclin:cdk complexes and negatively regulated by cyclin-dependent kinase inhibitors, including p21Cip1. Here we investigate the mechanisms regulating p21Cip1 levels in VSMCs and its role in controlling VSMC proliferation. METHODS AND RESULTS: We studied the S-phase associated kinase protein-2 (Skp2), an F-box protein implicated in the ubiquitination of p21Cip1. Overexpression of wild-type Skp2 or dominant-negative Skp2 decreased or increased p21Cip1 levels, respectively. Interestingly, levels of endogenous p21Cip1 and Skp2 were both increased in a phosphoinositide PI 3 kinase-dependent manner in late G1 phase. Increased expression of p21Cip1 occurred despite significantly increased Skp2-mediated proteasomal degradation. To determine the role of p21Cip1 in regulating VSMC proliferation, we used adenovirus-mediated overexpression and small-interfering RNA to elevate or silence p21Cip1 expression, respectively. Overexpression of p21Cip1 significantly inhibited VSCM proliferation. p21Cip1 silencing also inhibited proliferation and increased apoptotic cell death. CONCLUSIONS: Taken together, this data demonstrates that a balance between PI 3-kinase-driven upregulation and Skp2 mediated degradation controls the level of p21Cip1, which regulates VSMC proliferation in a biphasic manner. Low levels of p21Cip1 are also essential to counter apoptosis during cell-cycle progression. PMID- 16212952 TI - Vasculoprotective effect of insulin in the ischemic/reperfused canine heart: role of Akt-stimulated NO production. AB - OBJECTIVES: The objectives of this study were to investigate the vasculoprotective effects of glucose-insulin-potassium (GIK) on ischemia/reperfusion-induced coronary endothelial functional injury and to elucidate the mechanism involved. METHODS: Dogs were subjected to 50 min of coronary occlusion and 4 h of reperfusion. Vehicle, GIK, or GK were intravenously infused 5 min before reperfusion, and the coronary vascular dysfunction and endothelial apoptosis were determined. In a separate study, cultured endothelial cells were subjected to simulated ischemia/reperfusion, and the signaling pathway involved in insulin's anti-apoptotic effect was investigated. RESULTS: In vivo ischemia/reperfusion caused significant coronary vascular endothelial dysfunction as evidenced by reduced endothelium-dependent vasorelaxation, decreased nitric oxide (NO) production, and endothelial cell apoptosis as determined by caspase 3 activation and TUNEL staining. Treatment with GIK, but not GK, markedly improved the endothelium-dependent coronary vasorelaxation (P<0.01 versus vehicle), increased total NO production (P<0.01), and attenuated endothelial apoptosis. In cultured endothelial cells, treatment with insulin also markedly increased NO production and reduced simulated ischemia/reperfusion-induced apoptosis. Moreover, pre-treatment with either Akt inhibitor or NO synthase inhibitor almost abolished the anti-apoptotic effect exerted by insulin but not by SNAP, an NO donor. CONCLUSION: These results demonstrate that in vivo treatment with GIK at reperfusion attenuates ischemia/reperfusion-induced coronary endothelial dysfunction and endothelial apoptosis in an Akt-dependent and NO-mediated fashion. The coronary vasculoprotective effect elicited by insulin may contribute to the previously observed cardiac protective effect of GIK. PMID- 16212953 TI - A computer program for the simulation of fiber deposition in the human respiratory tract. AB - As inhaled fibers may lead to a variety of lung diseases, detailed information on their deposition in the human respiratory tract is an indispensable requirement in medical science. In the work presented here, a Visual Basic((R)) computer program, termed FIBROS, is described which enables the simulation of fibrous particle deposition in both the extrathoracic region and different parts of the lung itself, including the results of published numerical studies on inertial/interceptional as well as diffusional and gravitational deposition. The input window of FIBROS includes the selection of specific breathing conditions by variation of the tidal volume and breathing cycle. Furthermore, the user is able to determine fiber properties such as diameter, aspect ratio, specific weight, and fiber orientation with respect to the air stream in the upper and lower airways of the lungs. Besides the offer of various deposition formulae for each region of the respiratory tract, thereby also allowing a distinction between mouth and nose breathing, the user may select between different morphometric datasets of the lung and respective airway scaling procedures. Analysis routines of FIBROS include the estimation of regional deposition fractions, thereby distinguishing between extrathoracic, bronchial, and acinar compartments, and a calculation of generation-by-generation deposition probabilities within tubular and alveolar structures. Preliminary results presented here should demonstrate the effects on fiber deposition due to variations of the breathing behaviour and the particle properties. PMID- 16212954 TI - Intrathecally-administered histamine facilitates nociception through tachykinin NK1 and histamine H1 receptors: a study in histidine decarboxylase gene knockout mice. AB - Intrathecal injection of histamine elicited behavioral responses consisting of scratching, biting and licking in conscious mice. To study the participation of histamine in pain perception, histidine decarboxylase knockout mice were examined for pain threshold by means of three different kinds of noxious stimuli: thermal nociception (hot-plate, tail-flick, and paw-withdrawal), mechanical nociception (tail-pressure), and chemical nociception (formalin test and capsaicin test). Mutant mice lacking histidine decarboxylase showed significantly fewer nociceptive responses to the hot-plate, tail-flick, paw-withdrawal, tail pressure, formalin and capsaicin tests. Sensitivity to noxious stimuli in the histidine decarboxylase knockout mice was significantly lower when compared to the wild-type mice. The intrathecally-administered histamine (400 pmol) significantly shortened the latency in the histidine decarboxylase knockout mice, but not in the wild-type mice in tail-flick tests. Pyrilamine, a histamine H(1) receptor antagonist, but not ranitidine, a histamine H(2) receptor antagonist, produced inhibition of the induced behavioral responses in the tail-flick test when co-administered with histamine. Sendide, a tachykinin NK(1) receptor antagonist, inhibited histamine-induced nociceptive behavior in the histidine decarboxylase knockout mice. In contrast, the treatment with D-(-)-2 amino-5 phosponovaleric acid (D-APV), an N-methyl-D-aspartate (NMDA) receptor antagonist, did not prevent the induction of the behavioral responses by histamine. These studies substantiate the evidence that nociceptive behavior induced by intrathecal injection of histamine is largely mediated through tachykinin NK(1) and histamine H(1) receptors in the spinal cord. PMID- 16212955 TI - Leishmanin skin test in guinea pig with a single purified protein of Leishmania major. AB - A series of hybridomas was produced by fusion of SP2/0 myeloma cells with spleen cells of mice immunized with Leishmania major (L. major). The reactivity of secreted monoclonal antibodies (mAbs) was evaluated against available leishmanin antigen by enzyme linked immunosorbent assay. Only one hybridoma designated as 7F9 secreted IgG1 mAb which was shown to be reactive with leishmanin. This mAb was further tested against four species of Leishmania (L. donovani, L. tropica, L. infantum, L. major) and a recombinant gp63. Among the four species tested it was shown to be only reactive with promastigotes of L. major. The antigen recognized by this mAb was purified and analyzed from both sonicated and supernatant cultures of L. major by immunoaffinity chromatography and reverse phase high performance liquid chromatography. The purified antigen, which gave a single band of 56kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis elicited a strong delayed-type hypersensitivity (DTH) reaction in guinea pigs sensitized with L. major. It was almost of the same degree as that produced by leishmanin. These results suggest that an L. major-specific antigen is an alternative as a specific diagnostic skin test reagent, which could lead to a better understanding of the mechanism of DTH in L. major. PMID- 16212956 TI - Schistosoma japonicum: a method for transformation by electroporation. AB - Despite of our knowledge of genetic make up of schistosomes, a number of genes have not been characterized largely due to lack of effective transformation protocols. Here we present electroporation as a strategy for effective introduction of plasmids DNA into schistosomula and adults. Using plasmids of pEGFP-C1 as an expression vector, we first verified that the CMV promoter could direct EGFP to express in primary culture cells from Schistosoma japonicum. Subsequently, the plasmids were introduced into schistosomula and adults by electroporation and EGFP expression was demonstrated using molecular and microscopical methods. Our findings indicate that electroporation is an effective method for transformation of S. japonicum. PMID- 16212957 TI - Elucidation of subfamily segregation and intramolecular coevolution of the olfactomedin-like proteins by comprehensive phylogenetic analysis and gene expression pattern assessment. AB - The categorization of genes by structural distinctions relevant to biological characteristics is very important for understanding of gene functions and predicting functional implications of uncharacterized genes. It was absolutely necessary to deploy an effective and efficient strategy to deal with the complexity of the large olfactomedin-like (OLF) gene family sharing sequence similarity but playing diversified roles in many important biological processes, as the simple highest-hit homology analysis gave incomprehensive results and led to inappropriate annotation for some uncharacterized OLF members. In light of evolutionary information that may facilitate the classification of the OLF family and proper association of novel OLF genes with characterized homologs, we performed phylogenetic analysis on all 116 OLF proteins currently available, including two novel members cloned by our group. The OLF family segregated into seven subfamilies and members with similar domain compositions or functional properties all fell into relevant subfamilies. Furthermore, our Northern blot analysis and previous studies revealed that the typical human OLF members in each subfamily exhibited tissue-specific expression patterns, which in turn supported the segregation of the OLF subfamilies with functional divergence. Interestingly, the phylogenetic tree topology for the OLF domains alone was almost identical with that of the full-length tree representing the unique phylogenetic feature of full-length OLF proteins and their particular domain compositions. Moreover, each of the major functional domains of OLF proteins kept the same phylogenetic feature in defining similar topology of the tree. It indicates that the OLF domain and the various domains in flanking non-OLF regions have coevolved and are likely to be functionally interdependent. Expanded by a plausible gene duplication and domain couplings scenario, the OLF family comprises seven evolutionarily and functionally distinct subfamilies, in which each member shares similar structural and functional characteristics including the composition of coevolved and interdependent domains. The phylogenetically classified and preliminarily assessed subfamily framework may greatly facilitate the studying on the OLF proteins. Furthermore, it also demonstrated a feasible and reliable strategy to categorize novel genes and predict the functional implications of uncharacterized proteins based on the comprehensive phylogenetic classification of the subfamilies and their relevance to preliminary functional characteristics. PMID- 16212958 TI - Characterization of the DNA-binding site in the ferric uptake regulator protein from Escherichia coli by UV crosslinking and mass spectrometry. AB - Ferric uptake regulator protein (Fur) is activated by its cofactor iron to a state that binds to a specific DNA sequence called 'Fur box'. Using mass spectrometry-based methods, we showed that Tyr 55 of Escherichia coli Fur, as well as the two thymines in positions 18 and 19 of the consensus Fur Box, are involved with binding. A conformational model of the Fur-DNA complex is proposed, in which DNA is in contact with each H4 [A52-A64] Fur helix. We propose that this interaction is a common feature for the Fur-like proteins, such as Zur and PerR, and their respective DNA boxes. PMID- 16212959 TI - RACK1 mRNA translation is regulated via a rapamycin-sensitive pathway and coordinated with ribosomal protein synthesis. AB - RACK1 has been shown to interact with several proteins, this suggesting that it may play a central role in cell growth regulation. Some recent articles have described RACK1 as a component of the small ribosomal subunit. To investigate the relationship between RACK1 and ribosome, we analyzed RACK1 mRNA structure and regulation. Translational regulation was studied in HeLa cells subjected to serum or amino acid deprivation and stimulation. The results show that RACK1 mRNA has a 5' terminal oligopyrimidine sequence and that its translation is dependent on the availability of serum and amino acids in exactly the same way as any other vertebrate ribosomal protein mRNA. PMID- 16212960 TI - Identification of plasma membrane associated mature beta-hexosaminidase A, active towards GM2 ganglioside, in human fibroblasts. AB - Mature beta-hexosaminidase A has been found associated to the external leaflet of plasma membrane of cultured fibroblasts. The plasma membrane association of beta hexosaminidase A has been directly determined by cell surface biotinylation followed by affinity chromatography purification of the biotinylated proteins, and by immunocytochemistry. The immunological and biochemical characterization of biotinylated beta-hexosaminidase A revealed that the plasma membrane associated enzyme is fully processed, suggesting its lysosomal origin. PMID- 16212961 TI - Analysis of glycoside hydrolase family 98: catalytic machinery, mechanism and a novel putative carbohydrate binding module. AB - Glycoside hydrolases (GHs) are diverse enzymes of biotechnological and medical importance. Bioinformatics contributes to our understanding of GH structure and function in various ways, including dissection of their typically modular structures and detection of the distant evolutionary relationships between families that often allow for prediction of catalytic sites. Here these twin strands are applied to the recently described GH98 family, the founder member of which is a blood group glycotope-cleaving endo-beta-galactosidase of potential medical importance from Clostridium perfringens. Three domains can be discerned including a central catalytic TIM barrel domain in which putative catalytic residues can be assigned. Distant homologies and domain contexts suggest that the N-terminal domain is a novel carbohydrate binding module. PMID- 16212962 TI - HIPK2 inhibits both MDM2 gene and protein by, respectively, p53-dependent and independent regulations. AB - We address here the involvement of the homeodomain-interacting protein kinase 2 (HIPK2)/p53 complex on MDM2 regulation following apoptotic DNA damage. Our results provide a plausible transcriptional (p53-dependent) and posttranscriptional (p53-independent) double mechanism by which HIPK2 accomplishes MDM2 downmodulation. First, in wtp53-carrying cells HIPK2-dependent p53Ser46 phosphorylation selectively inhibits MDM2 at transcriptional level. Secondly, HIPK2 interacts with MDM2 in vitro and in vivo and promotes MDM2 nuclear export and proteasomal degradation, in p53-null cellular context. This p53-independent effect is likely mediated by HIPK2 catalytic activity and we found that HIPK2 phosphorylates MDM2 in vitro. In response to DNA damage, depletion of HIPK2 by RNA-interference abolishes MDM2 protein degradation. We propose that HIPK2 contributes to drug-induced modulation of MDM2 activity at transcriptional (through p53Ser46 phosphorylation) and posttranscriptional (through p53-independent subcellular re-localization and proteasomal degradation) levels. PMID- 16212963 TI - Natriuretic peptides are negative modulators of adrenocortical cell function of the eastern fence lizard (Sceloporus undulatus). AB - Elucidation of the role of natriuretic peptides (NPs) in vertebrate adrenal steroidogenesis has been facilitated by the use of freshly dispersed adrenocortical cells. Our recent characterization of lizard adrenocortical cells [Carsia, R.V., John-Alder, H.B., 2003. Seasonal alterations in adrenocortical cell function associated with stress-responsiveness and sex in the Eastern Fence Lizard (Sceloporus undulatus). Horm. Behav. 43, 408-420] provided the opportunity to examine the influence of atrial natriuretic peptides (ANPs) and related NPs on reptilian adrenal steroidogenesis at the cellular level. In the present report, the action of NPs on lizard adrenal steroidogenesis was investigated using freshly dispersed adrenocortical cells derived from the Eastern Fence Lizard (Sceloporus undulatus). Basal production rates of aldosterone and corticosterone and maximal angiotensin II (ANG II)-induced production rates of these corticosteroids were inhibited with high efficacy (75-90%) by rat ANP at potencies of 0.4-0.7 nM. By contrast, rat ANP had no effect on maximal production rates of these corticosteroids in response to a maximal steroidogenic concentration of adrenocorticotropin (ACTH; 1 nM). However, rat ANP inhibited aldosterone and corticosterone production rates in response to a half-maximal steroidogenic concentration of ACTH (10 pM; approximately 50 pg/ml), albeit with less efficacy ( approximately 50%) and potency (approximately 6 nM) than for ANG II. Rat and eel ANP and rat and chicken brain natriuretic peptide (BNP) were equally efficacious at inhibiting maximal ANG II-induced aldosterone and corticosterone production but with different potencies. The order of inhibitory potency was rat ANP = chicken BNP > eel ANP > rat BNP. However, a specific peptide ligand for the NP clearance receptor was without effect. This study indicates that ANP and related NPs are efficacious inhibitors of lizard adrenal steroidogenesis by acting directly at the level of the adrenocortical cell. PMID- 16212967 TI - Normalizing temporal patterns to analyze sit-to-stand movements by using registration of functional data. AB - Functional data analysis techniques provide an alternative way of representing movement and movement variability as a function of time. In particular, the registration of functional data provides a local normalization of time functions. This normalization transforms a set of curves, records of repeated trials, yielding a new set of curves that only vary in terms of amplitude. Therefore, main events occur at the "same time" for all transformed curves and interesting features of individual recordings remain after averaging processes. This paper presents an application of the registration process to the analysis of the vertical forces exerted on the ground by both feet during the sit-to-stand movement. This movement is particularly interesting in functional evaluations related to balance control, lower extremity dysfunction or low-back pain. PMID- 16212965 TI - Overexpression of COX-2, Prostaglandin E synthase-1 and prostaglandin E receptors in blood mononuclear cells and plaque of patients with carotid atherosclerosis: regulation by nuclear factor-kappaB. AB - BACKGROUND AND OBJECTIVE: Prostaglandin E2 (PGE(2), a product of the cyclooxygenase 2 (COX-2) and membrane-associated Prostaglandin E Synthase (mPGES 1) pathway, has been implicated in the instability of atherosclerotic plaques. We have studied COX-2, mPGES-1 and PGE2 receptors (EPs) expression in peripheral blood mononuclear cells (PBMC) and atherosclerotic plaques of 29 patients with carotid stenosis as well as the effect of different nuclear factor-kappaB (NF kappaB) inhibitors on COX-2, mPGES-1 and EPs expression in cultured monocytic cells (THP-1). METHODS: COX-2, mPGES-1 and EP expression was analyzed by RT-PCR (PBMC), immunohistochemistry (plaques) and Western blot (THP-1). PGE2 levels were determined by ELISA (plasma and cell supernatants). RESULTS: In relation to healthy controls, COX-2, mPGES-1 and EP-3/EP-4 mRNA expression was increased in PBMC from patients. In the inflammatory region of atherosclerotic plaques, an increase of COX-2, mPGES-1 and EPs expression was also observed. Activated NF kappaB and COX-2, mPGES-1 and EPs proteins were colocalized in the plaque's cells. In cytokine-treated cultured THP-1, the NF-kappaB inhibitors parthenolide, Bay 11-7082 and PDTC reduced COX-2, mPGES-1 and EP-1/EP-3/EP-4 expression as well as PGE2 levels. By employing specific agonists and antagonists, we noted that the cytokine- and PGE2-induced metalloproteinase 9 (MMP-9) expression and activity occurs through EP-1/EP-3/EP-4, an effect downregulated by NF-kappaB inhibitors. CONCLUSIONS: Patients with carotid atherosclerosis depict an overexpression of COX-2, mPGES-1 and EPs simultaneously in the PBMC as well as in the vulnerable region of plaques. The studies in cultured monocytic cells suggest that NF-kappaB inhibitors and/or EPs antagonists could represent a novel therapeutic approach to the treatment of plaque instability and rupture. PMID- 16212968 TI - Classification of gait patterns in the time-frequency domain. AB - This paper describes the classification of gait patterns among descending stairs, ascending stairs and level walking activities using accelerometers arranged in antero-posterior and vertical direction on the shoulder of a garment. Gait patterns in continuous accelerometer records were classified in two steps. In the first step, direct spatial correlation of discrete dyadic wavelet coefficients was applied to separate the segments of gait patterns in the continuous accelerometer record. Compared to the reference system, averaged absolute error 0.387 s for ascending stairs and 0.404 s for descending stairs were achieved. The overall sensitivity and specificity of ascending stairs were 98.79% and 99.52%, and those of descending stairs were 97.35% and 99.62%. In the second step, powers of wavelet coefficients of 2 s time duration from separated segments of vertical and antero-posterior acceleration signals were used as features in classification. Our results proved a reliable technique of measuring gait patterns during physical activity. PMID- 16212969 TI - Real-time subject-specific monitoring of internal deformations and stresses in the soft tissues of the foot: a new approach in gait analysis. AB - No technology is presently available to provide real-time information on internal deformations and stresses in plantar soft tissues of individuals during evaluation of the gait pattern. Because internal deformations and stresses in the plantar pad are critical factors in foot injuries such as diabetic foot ulceration, this severely limits evaluation of patients. To allow such real-time subject-specific analysis, we developed a hierarchal modeling system which integrates a two-dimensional gross structural model of the foot (high-order model) with local finite element (FE) models of the plantar tissue padding the calcaneus and medial metatarsal heads (low-order models). The high-order whole foot model provides real-time analytical evaluations of the time-dependent plantar fascia tensile forces during the stance phase. These force evaluations are transferred, together with foot-shoe local reaction forces, also measured in real time (under the calcaneus, medial metatarsals and hallux), to the low-order FE models of the plantar pad, where they serve as boundary conditions for analyses of local deformations and stresses in the plantar pad. After careful verification of our custom-made FE solver and of our foot model system with respect to previous literature and against experimental results from a synthetic foot phantom, we conducted human studies in which plantar tissue loading was evaluated in real time during treadmill gait in healthy individuals (N = 4). We concluded that internal deformations and stresses in the plantar pad during gait cannot be predicted from merely measuring the foot-shoe force reactions. Internal loading of the plantar pad is constituted by a complex interaction between the anatomical structure and mechanical behavior of the foot skeleton and soft tissues, the body characteristics, the gait pattern and footwear. Real-time FE monitoring of internal deformations and stresses in the plantar pad is therefore required to identify elevated deformation/stress exposures toward utilizing it in gait laboratories to protect feet that are susceptible to injury. PMID- 16212970 TI - Analysis of triazolopyrimidine herbicides in soils using field-enhanced sample injection-coelectroosmotic capillary electrophoresis combined with solid-phase extraction. AB - In this work, a combined methodology using off-line solid-phase extraction (SPE), on-line field-enhanced sample injection (FESI) and coelectroosmotic capillary electrophoresis with UV detection (CE-UV) is developed for the trace analysis of five triazolopyrimidine sulfonanilide pesticides (i.e., flumetsulam, florasulam, cloransulam-methyl, diclosulam and metosulam). An adequate background electrolyte (BGE) was obtained for the separation of these pesticides using hexadimethrine bromide (HDB) as electroosmotic flow (EOF) modifier. This BGE consisted of 0.00042% HDB, 11 mM formic acid, 16 mM ammonium carbonate and 2.5 mM alpha-CD solution at pH 7.6. The use of this running buffer together with the FESI preconcentration method provided limits of detection (LODs) in the low microg/L range (i.e., between 13.0 and 31.5 microg/L). The optimized FESI-CE-UV method was combined with off-line SPE using C(18) cartridges and applied to the determination of the selected group of pesticides in soil samples. Recovery percentages ranged between 50 and 84% in these samples with LODs between 18 and 34 microg/kg. This work shows the great possibilities of the combined use of SPE FESI-CE-UV to improve CE sensitivity allowing the achievement of LODs similar to other analytical techniques as GC or HPLC. PMID- 16212971 TI - Field-amplified sample injection and in-capillary derivatization for sensitivity improvement of the electrophoretic determination of histamine. AB - The feasibility of the combination of field-amplified sample injection (FASI) and in-capillary derivatization was explored for improving sensitivity of histamine in capillary electrophoresis (CE). Naphthalene-2,3-dicarboxaldehyde (NDA) was used as derivatization reagent. The reagent and sample was introduced by tandem mode. The derivatization was accomplished by at-inlet mode with standing time of 1.5 min. The combination of FASI and in-capillary derivatization was successfully achieved with about 400-fold concentration sensitivity enhancement compared to pre-capillary derivatization at the same set-up. The detection limit of concentration for histamine reached 1.25 x 10(-11) M by CE and fluorescence detection with S/N = 3. Parameters affecting FASI and in-capillary derivatization process including sample matrix, buffer concentration and reagent injection amount, were investigated. PMID- 16212972 TI - Nano-fibriform production of silica from natural chrysotile. AB - Nano-fibriform silica was extracted from chrysotile by the acid-leaching method. The acid-leached residue of chrysotile has been studied by TEM, XRD, FT-IR, and thermal analysis techniques, etc. When the magnesium leaching degree (MLD) is over 90%, the nano-fibriform silica consists of hydrous silicon dioxide (above 90%) with small amount of magnesium trapped inside the SiO network. The amount of hydroxyl on surface of nano-fibriform silica is 6 unit nm(-2). This value is between the values of fumed and precipitated silica. This study shows that nano fibriform silica is a kind of amorphous matter with a high special surface area (368 m2/g), a high adsorption (330 cm3/g), and a larger pore volume (0.51 cm3/g). The diameter of a single silica fiber is 20-30 nm. The nitrogen adsorption isotherm is similar to Type IV curve. The nano-fibriform silica is one of mesopores materials. PMID- 16212973 TI - Coadsorption of Cd(II) and oxalate ions at the TiO2/electrolyte solution interface. AB - The study of the adsorptions of cadmium and oxalate ions at the titania/electrolyte interface and the changes of the electrical double layer (edl) structure in this system are presented. The adsorption of cadmium or oxalate ions was calculated from an uptake of their concentration from the solution. The concentration of Cd(II) or oxalate ions in the solution was determined by radiotracer method. For labeling the solution 14C and 115Cd isotopes were used. Coadsorption of Cd(II) and oxalic ions was determined simultaneously. Besides, the main properties of the edl, i.e., surface charge density and zeta potential were determined by potentiometer titration and electrophoresis measurements, respectively. The adsorption of cadmium ions increases with pH increase and shifts with an increase of the initial concentration of Cd(II) ions towards higher pH values. The adsorption process causes an increase of negatively charged sites on anatase and a decrease of the zeta potential with an increase of initial concentration of these ions. The adsorption of oxalate anions at the titania/electrolyte interface proceeds through the exchange with hydroxyl groups. A decrease of pH produces an increase of adsorption of oxalate ions. The processes of anion adsorption lead to increase the number of the positively charged sites at the titania surface. However, specific adsorption of bidenate ligand as oxalate on one surface hydroxyl group may form inner sphere complexes on the metal oxide surface and may overcharge the compact part of the edl. The presence of oxalate ions in the system affects the adsorption of Cd(II) ions on TiO2, increasing the adsorption at low pH range and decreasing the adsorption at high pH range. Using adsorption as a function of pH data, some characteristic parameters of adsorption envelope were calculated. PMID- 16212974 TI - Effect of a neutral water-soluble polymer on the lamellar phase of a zwitterionic surfactant system. AB - We have studied the effect of adding a water-soluble polymers (PEG) to the lamellar phases of the ternary system tetradecyldimethylaminoxide (C14DMAO) hexanol-water. The results of Freeze-Fracture Electron Microscopy (FFEM) and Small Angle X-ray Scattering (SAXS) experiments show that the addition of the polymer induces the spontaneous formation of highly monodisperse multilayered vesicles above a threshold polymer concentration. PMID- 16212975 TI - Development of in-situ hybridization for the detection of Mycoplasma haemosuis (Eperythrozoon suis) in formalin-fixed, paraffin wax-embedded tissues from experimentally infected splenectomized pigs. AB - Mycoplasma haemosuis DNA was detected in experimentally infected splenectomized pigs by in-situ hybridization (ISH) with a nonradioactive digoxigenin-labelled DNA probe. An 839 base pair DNA probe targeting a 16S rRNA gene was generated by the polymerase chain reaction. Eight 6-week-old pigs were inoculated intraperitoneally with 6 ml of M. haemosuis-infected pig blood and eight negative control pigs were inoculated intraperitoneally with 6 ml of M. haemosuis-free blood. Two pigs from each group were killed for examination at 3, 7, 15 and 30 days post-inoculation (dpi). Red blood cells infected with M. haemosuis were first detected by light microscopy at 3 to 7 dpi. No M. haemosuis was observed in negative control pigs. Hybridization signals were evident in blood from the infected pigs at 3 dpi. The ISH method developed in this study was useful for the detection of M. haemosuis DNA in formalin-fixed, paraffin wax-embedded tissues and may be valuable for studying the pathogenesis of M. haemosuis infection. PMID- 16212976 TI - Fluorescence based oxygen uptake analysis in the study of metabolic responses to apoptosis induction. AB - Mitochondrial activity has been shown to be centrally involved in the progression of apoptosis. The electron transport chain is a major player in this process and oxygen uptake analysis provides detailed information on its activity. Here we examined the ability of a fluorescence based oxygen uptake assay to inform on cellular responses to apoptosis induction. HL60 cells treated with camptothecin and UV light were used as a model and the ability of the assay to detect dose and time dependent decreases in respiratory activity analysed. The data obtained were compared to more specific markers of apoptosis including annexin V binding, and caspase-3 activity. Reductions in oxygen uptake rates were seen at lower doses than increases in annexin V binding or mitochondrial membrane potential depolarisation. These reductions were observed earlier than detectable caspase-3 activity and were unaffected by pre-treatment with the caspase-3 inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoro-methylketone (zVADfmk). PMID- 16212977 TI - Production and characterization of mouse ureteric bud cell-specific rat hybridoma antibodies utilizing subtractive immunization and high-throughput screening. AB - The highly branched collecting system of the kidney arises developmentally from the ureteric bud (UB) by a process of branching morphogenesis. This process is critical for the normal development of the collecting ducts and pelvis of the kidney, and is tightly controlled by the spatial and temporal expression of numerous proteins. To identify cell proteins that are differentially expressed by the UB relative to those expressed by the highly differentiated collecting ducts of the adult kidney, two mouse cell populations derived from either the early UB or the adult inner medullary collecting duct (IMCD) were used. A subtractive immunization strategy was performed in rats to generate monoclonal antibodies that preferentially reacted with antigens on UB, but not IMCD cells. In addition, the technique of antibody printing, a novel high-throughput antibody screening method for determining the specificities of a large number of monoclonal antibodies, is described. The methodologies outlined in this manuscript have broad applicability as they demonstrate that subtractive immunization can be performed in rats with cells derived from mice. Additionally, the high-throughput screening methods should facilitate the use of subtractive immunization for identifying antibodies that can distinguish differences in proteins expressed in closely related cell types. PMID- 16212978 TI - Enzyme-substrate complex structures of a GH39 beta-xylosidase from Geobacillus stearothermophilus. AB - Beta-D-Xylosidases are glycoside hydrolases that catalyse the release of xylose units from short xylooligosaccharides and are engaged in the final breakdown of plant cell-wall hemicelluloses. beta-D-Xylosidases are found in glycoside hydrolase families 3, 39, 43, 52 and 54. The first crystal structure of a GH39 beta-xylosidase revealed a multi-domain organization with the catalytic domain having the canonical (beta/alpha)8 barrel fold. Here, we report the crystal structure of the GH39 Geobacillus stearothermophilus beta-D-xylosidase, inactivated by a point mutation of the general acid-base residue E160A, in complex with the chromogenic substrate molecule 2,5-dinitrophenyl-beta-D xyloside. Surprisingly, six of the eight active sites present in the crystallographic asymmetric unit contain the trapped covalent glycosyl-enzyme intermediate, while two of them still contain the uncleaved substrate. The structural characterization of these two critical species along the reaction coordinate of this enzyme identifies the residues forming its xyloside-binding pocket as well as those essential for its aglycone recognition. PMID- 16212979 TI - Clinical and radiographic subtypes of vascular cognitive impairment in a clinic based cohort study. AB - BACKGROUND AND PURPOSE: There is a need for empirical studies to define criteria for vascular cognitive impairment (VCI) subtypes. In this paper, we report the predictive validity of a subtype classification scheme based on clinical and radiographic features. METHODS: Nine Canadian memory clinics participated in the Consortium to Investigate Vascular Impairment of Cognition. This cohort consisted of 1347 patients, of whom 324 had VCI, and was followed for up to 30 months. RESULTS: Clinical and neuroimaging features defined three subtypes: vascular cognitive impairment, no dementia, (n=97), vascular dementia (n=101) and mixed neurodegenerative/vascular dementia (n=126). Any ischemic lesion on neuroimaging increased the odds (odds ratio=9.31; 95% confidence interval 6.46, 13.39) of a VCI diagnosis. No VCI subtype, however, was associated with a specific neuroimaging abnormality. Compared to those with no cognitive impairment, patients with each VCI subtype had higher rates of death and institutionalization (hazard ratio for combined adverse events=6.08, p<0.001). CONCLUSIONS: Both clinical features and radiographic features help establish a diagnosis of VCI. The outcomes of VCI subtypes, however, are more strongly associated with clinical features than with radiographic ones. PMID- 16212981 TI - An evolutionary relationship between genetic variation and phenotypic fluctuation. AB - The relevance of phenotype fluctuations among clones (i.e., organisms with identical genes) to evolution has recently been recognized both theoretically and experimentally. By considering the stability of the distributions of genetic variations and phenotype fluctuations, we derive a general inequality between the phenotype variance due to genetic differences and the intrinsic phenotype variance of clones. For a given mutation rate, an approximately linear relationship between the two is obtained which elucidates the consistency between the fundamental theorem of natural selection by Fisher and the evolutionary fluctuation-response relationship (fluctuation dissipation theorem) proposed recently. A general condition for the error catastrophe is also derived as the violation of the inequality, which sets up the limit to the speed of stable evolution. All of these theoretical results are confirmed by a numerical evolution experiment of a cell that consists of a catalytic reaction network. Based on the relationships proposed here, relevance of the phenotypic plasticity to evolution as well as the genetic assimilation is discussed. PMID- 16212980 TI - Over-expression of p73beta results in apoptotic death of post-mitotic hNT neurons. AB - The p53-related p73 protein is an important mediator of apoptosis, development and tumorigenesis. Previously, we showed that over-expression of the p73beta isoform induced apoptosis in proliferating neuronal cells; however, the study did not address the effect of p73 in post-mitotic neurons. To address this question, we used post-mitotic hNT neurons, which have been used as a model of human central nervous system neurons. We found that over-expression of p73beta in hNT neurons resulted in apoptosis and an increase in the expression of p57Kip2 and Bax, but no increase in p53 expression. These results suggest that apoptosis of post-mitotic neurons induced by p73beta may involve these mediators. Understanding the regulation of p73 expression will be important for understanding the development of the nervous system and may have implications for the treatment of neurological diseases. PMID- 16212983 TI - Divergent role of calcium on Abeta- and MPTP-induced cell death in SK-N-SH neuroblastoma. AB - We attempted to clarify the role of Ca2+ in cell death caused by beta-amyloid protein (Abeta) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in SK-N SH neuroblastoma, respectively. Two insults both reduced cell viability in a concentration-dependent manner and induced equal cytotoxicity in the presence of 20 microM Abeta and 0.4 mM MPTP for 72 h, respectively (68+/-7 vs. 64+/-6% viability). Time-related study showed that Abeta evoked cell death occurred quickly at 24 h. Relatively, MPTP exhibited a delayed cell death significantly after 72 h of culture. Pretreating the cells with nimodipine and chelating of Ca2+ by EGTA plus 1,2-bis-(O-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM) successfully rescued Abeta-induced cell death but failed to prevent MPTP toxicity. ELISA determination of mono/oligonucleosomes accumulation showed the mode of cell death evoked by MPTP was presumably apoptosis while by Abeta was necrosis. SK-N-SH cells constitutively expressed the alpha(1C) subunit of L-type Ca2+ channel and exposure to Abeta or MPTP for 96 h did not further modify its expression. By contrast, alpha(1D) subunit was undetectable or low level expressed in basal condition, but was induced to express after Abeta and MPTP stimulation in a time-dependent manner. Functional assay revealed that KCl-evoked [Ca2+]i rise was significantly greater in Abeta-, but not in MPTP-treated cells when compared with control. Taken together, these results showed that Abeta and MPTP elicited different mode of cell death in SK-N SH. Nevertheless, Ca2+ overload seems to solely display a crucial role in Abeta induced cytotoxicity and over-expressed alpha(1D) may contribute to the disruption of cellular Ca2+ homeostasis. PMID- 16212982 TI - Hypertension induced by nitric oxide synthase inhibitor increases responsiveness of ventricular myocardium and aorta of rat tissue to adrenomedullin stimulation in vitro. AB - In this work, we aimed to observe the changes in adrenomedullin (ADM) and its receptor-calcitonin receptor-like receptor (CL), receptor activity-modifying protein (RAMP) 1, RAMP2 and RAMP3-in cardiac ventricles and aortas of hypertensive rats, and the responsiveness of injured cardiovascular tissue to ADM, then to illustrate the protective mechanism of ADM on the cardiovascular system. Male SD rats were subjected to treatment with chronic N(G)-nitro-L arginine (L-NNA), an inhibitor of nitric oxide synthase. The ADM contents and cAMP production in myocardia and aortas were measured by RIA. The mRNA levels of ADM, CL, and RAMP1-3 were determined by RT-PCR. L-NNA induced severe hypertension and cardiomegaly. The ir-ADM content in plasma, ventricles and aortas in L-NNA treated animals increased by 80%, 72% and 57% (all p<0.01), respectively. Furthermore, mRNA levels of ADM, CL, RAMP2 and RAMP3 were elevated by 91%, 33%, 50% and 72.5% (all p<0.01), respectively, in ventricles and by 95%, 177%, 74.7% and 85% (all p<0.01), respectively, in aortas. mRNA level of RAMP1 was elevated by 129% (p<0.01) in aortas but no significant difference in ventricles. The elevated mRNA levels of RAMP2 and RAMP3 were positively correlated with that of ADM in hypertrophic ventricles (r=0.633 and 0.828, p<0.01, respectively) and the elevated mRNA levels of CL, RAMP2 and RAMP3 were positively correlated with that of ADM in aortas (r=0.941, 0.943 and 0.736, all p<0.01, respectively). The response of ventricular myocardia and aortas to ADM administration potentiated, and the production of cAMP was increased by 41% and 68% (both p<0.01), respectively. ADM-stimulated cAMP generation in ventricular myocardia and aortas was blocked by administration of both ADM22-52, the specific antagonist of ADM receptor, and CGRP8-37, the antagonist of the CGRP1 receptor. The results showed an increased in cardiovascular ADM generation and an up-regulation of the gene expression of ADM and its receptor-CL, RAMP1-3 during hypertension, augmented responsiveness of ventricular myocardia and aortas of hypertensive rats to ADM, suggesting that these receptors may play a role in the cardiovascular adaptation in response to sub-chronic NO-inhibition. PMID- 16212984 TI - PCBs and PAHs in sea-surface microlayer and sub-surface water samples of the Venice Lagoon (Italy). AB - Polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) are two classes of micropollutants intensively monitored and regulated due to their toxicity, persistency and wide diffusion. Their concentrations have been investigated in sea-microlayer (SML) and sub-surface water (SSW) samples collected at two sites of the Venice Lagoon, a fragile ecosystem highly influenced by industrial and anthropogenic emissions. The total sigmaPCB concentration varies from 0.45 ng/l to 2.1 ng/l in SSW while a clear enrichment is observed in the SML, where it ranges from 1.2 ng/l to 10.5 ng/l. The total sigmaPAH concentration shows marked differences between the two stations and varies from 12.4 ng/l to 266.8 ng/l in SSW; in SML it is more uniform and ranges from 19.6 ng/l to 178.9 ng/l. The enrichment factors are not larger than 1 for both pollutants in the 'dissolved' phase, while they are most significant for the 'particulate' phase (sigmaPCB: 5-9; sigmaPAH: 4-14). PMID- 16212985 TI - Risk perception bias, self-reporting of illness, and the validity of reported results in an epidemiologic study of recreational water associated illnesses. AB - Epidemiologic studies of water associated illness often have to rely on self reported symptoms of the outcome illness(es) under study. Individual participant's perception of risk, in theory, can affect the validity of self reported symptoms. The magnitude and effect of possible "risk perception bias" was evaluated as part of a series of randomized trials designed to assess infectious disease transmission via exposure to marine recreational waters with modest sewage contamination. All study subjects were blinded to both their individual indices of exposure and the outcome illnesses under study. Of the five outcome illnesses studied, the effect of "risk perception bias" only effected one: skin ailments. Although analysis of crude rates of skin ailments showed the exposed group (bathers) to be 3.5 times more likely to report skin ailments relative to the non-exposed (non-bathers), when the data was stratified by any perceived health risk of bathing in such waters, this association was shown to be spurious in nature. Bathers having pre-conceived notions of any health risk due to the exposure were 10.63 times more likely to report skin ailments relative to the unexposed (non-bathers) (95% CI 2.36-47.8, P = 0.0002), while bathers without any pre-conceived notion of risk were no more likely to report skin ailments relative to non-bathers (OR = 0.60, 95% CI 0.11-3.24, P = 0.71). Further stratification by exposure grouping showed bathers with pre-conceived notions of excess risk to be 4.78 times more likely to report skin ailments relative to bathers without any notion of excess risk (95% CI 1.04-21.86, P = 0.03), while among non-bathers those with pre-conceived notions of risk were 3.70 times less likely to report skin ailments relative to non-bathers without any pre-conceived notion of risk (95% CI 0.70-19.60, P = 0.10). This study shows that "risk perception bias" can be strong enough to lead to spurious associations in the presence of self-reported symptoms, and should be controlled for in future epidemiologic studies of recreational water associated illnesses and other water associated environmental exposures where the use of self-reported symptoms cannot be avoided. PMID- 16212986 TI - The dependence of organohalogen compound concentrations on herring age and size in the Bothnian Sea, northern Baltic. AB - The concentrations and composition profiles of polychlorinated dibenzo-p-dioxins and dibenzofurans, polychlorinated biphenyls, polybrominated diphenyl ethers and polychlorinated naphthalenes were determined in herring individuals collected from the commercial catches of the Bothnian Sea, northern Baltic. The median age of herring was 5.0 years and in the muscle the median toxic equivalent concentration of PCDD/Fs was 5.6 WHO(PCDD/F)-TEQ pg/g fresh weight (fw) and that of PCBs 2.7 WHO(PCB)-TEQ pg/g fw. The median sum concentration of PBDEs was 1.4 ng/g fw and that of PCNs 0.1 ng/g fw. Differences in age-dependent accumulation between the organohalogen groups and individual congeners were major. In the Bothnian Sea the content of organohalogen compounds in herring is obviously elevated due to the availability and large proportion of Mysis crustaceans in their diet. More intensive fishing could reduce the concentrations of organohalogens, including the abundant, accumulative and toxic 2,3,4,7,8-PeCDF congener in the Baltic herring catch. PMID- 16212987 TI - Predicting summer hypoxia in the northern Gulf of Mexico: riverine N, P, and Si loading. AB - We conducted a statistical analysis to discern the relative strengths of the loading of various forms of nitrogen, phosphorus, dissolved silicate and their molar ratios on the variance in the size of the summertime low oxygen zone found off the Mississippi River, northern Gulf of Mexico. A stable statistical model that included Year and riverine nitrate+nitrite loading for the 2 months prior to the measurement of hypoxic zone size described 82% of its variation in size from 1978 to 2004. The usefulness of the term Year is consistent with the documented increase in carbon stored in sediments after the 1970s, which is when the hypoxic zone is predicted to have become a regular feature on the shelf and to have expanded westward. The increased carbon storage is anticipated to cause a sedimentary respiratory demand influencing the size of the zone, and whose temporal influence is cumulative and transcends the annual variations in nitrogen loading. The variable Year is negatively correlated with the TN:TP ratio in a way that suggests N, not P, has become more important as a factor limiting phytoplankton growth in the last 20 years. Nitrogen, in particular nitrate+nitrite, and not phosphorus, dissolved silicate, or their molar ratios, appears to be the major driving factor influencing the size of the hypoxic zone on this shelf. This conclusion is consistent with cross-system analyses that conclude that the TN:TP ratio in the Mississippi River, currently fluctuating around 20:1, is indicative of nitrogen, not phosphorus, limitation of phytoplankton growth. Nutrient management that places stronger emphasis on reducing nitrogen loading as compared to phosphorus loading, is justified. PMID- 16212988 TI - Distribution of manufactured floating marine debris in near-shore areas around Japan. PMID- 16212989 TI - Changes in octocoral communities and benthic cover along a water quality gradient in the reefs of Hong Kong. AB - Cover of the main reef benthic groups, and abundances and taxonomic richness of octocorals were surveyed in the reefs of Hong Kong, and related to spatial and water quality gradients. Nutrient and particle concentrations are high throughout the area, with concentrations declining from the south towards the north-eastern region. Regression tree analyses showed that hard coral cover was most strongly related to water clarity, that macroalgal cover was highest in areas with high wave action and high water clarity, and that crustose coralline algae were negatively related to sedimentation. Octocoral communities (42 species in 23 genera) were dominated by zooxanthellae-free taxa; those few species with zooxanthellae were restricted to reefs with low wave action and high water clarity in the north-eastern region. The water quality gradient spans from conditions that are marginal for zooxanthellate octocorals while still supporting diverse scleractinian communities, towards an estuarine endpoint where zooxanthellate octocorals cease to exist and hard coral communities are reduced to a few resilient colonies. The data suggest that the types, abundances and richness of zooxanthellate octocorals, and the shift from zooxanthellate to azooxanthellate octocoral communities, may act as useful indicators of water clarity in regions where long-term water quality data are unavailable. PMID- 16212990 TI - Expression of GFP-tagged neuronal glutamate transporters in cerebellar Purkinje neurons. AB - Of the five excitatory amino acid transporters (EAATs) identified, two genes are expressed by neurons (EAAT3 and EAAT4) and give rise to transporters confined to neuronal cell bodies and dendrites. At an ultrastructural level, EAAT3 and EAAT4 proteins are clustered at the edges of postsynaptic densities of excitatory synapses. This pattern of localization suggests that postsynaptic EAATs may help to limit spillover of glutamate from excitatory synapses. In an effort to study transporter localization in living neurons and ultimately to manipulate uptake at intact synapses, we have developed viral reagents encoding neuronal EAATs tagged with GFP. We demonstrate that these fusion proteins are capable of Na(+) dependent glutamate uptake, that they generate ionic conductances indistinguishable from their wild-type counterparts, and that GFP does not alter their glutamate dose-dependence. Two-photon microscopy was used to examine fusion protein expression in Purkinje neurons in acute cerebellar slices. Both EAAT3-GFP and EAAT4-GFP were observed at high levels in the dendritic spines of transfected Purkinje neurons. These findings indicate that functional EAAT fusion proteins can be synthesized and appropriately trafficked to postsynaptic compartments. Furthermore, they validate a powerful system for looking at EAAT function in situ. PMID- 16212991 TI - Rapid delivery of the dopamine transporter to the plasmalemmal membrane upon amphetamine stimulation. AB - The dopamine transporter, DAT, is a primary regulator of dopamine (DA) signaling at the synapse. Persistent stimulation with the substrate amphetamine (AMPH) promotes DAT internalization. AMPH rapidly elicits DA efflux, yet its effect on DAT trafficking at short times is unknown. We examined the rapid effect of AMPH on DAT trafficking in rat striatal synaptosomes using biotinylation to label surface DAT. Within 30s of treatment with 3 microM AMPH, synaptosomal DAT surface expression increased to 163% of control and remained elevated through at least 1 min before returning to control levels at 2.5 min. The increase in surface DAT was cocaine-sensitive but was not produced by DA itself. A 1-min preincubation with AMPH did not alter [(3)H]DA uptake, but did result in a higher basal DA efflux and efflux elicited in the presence of AMPH as compared to vehicle pretreatment. Reversible biotinylation experiments demonstrated that the AMPH stimulated rise in surface DAT is due to an increase in the delivery of DAT to the plasmalemmal membrane rather than a reduction of the endocytic process. These studies suggest that AMPH has a biphasic effect on DAT trafficking and acts rapidly to regulate DAT in the plasmalemmal membrane. PMID- 16212992 TI - Expression studies of naturally occurring human dopamine transporter variants identifies a novel state of transporter inactivation associated with Val382Ala. AB - Multiple, rare, human dopamine (DA) transporter (hDAT, SLC6A3) coding variants have been described, though to date they have been incompletely characterized. Here we present studies analyzing the function and regulation of five naturally occurring coding variants, V55A, R237Q, V382A, A559V and E602G, expressed in COS 7 and SH-SY5Y cells. All variants, except V382A, exhibited levels of surface protein expression and DA transport activity comparable to hDAT. V382A, divergent at the most highly conserved residue among reported hDAT variants, exhibited significantly diminished surface expression, likely derived from inefficient plasma membrane delivery. Moreover, a greater expression of V382A protein was required to achieve comparable levels of transport to hDAT, consistent with a loss of transport function. V382A displayed a decrease in sensitivity to phorbol ester (PMA)-induced internalization, as well as an altered substrate selectivity for DA versus norepinephrine (NE). Analysis of PMA-induced V382A internalization revealed a trafficking-independent action of PMA, consistent with the existence of a surface-localized, transport-inactive state. PMID- 16212994 TI - Audiogenic seizures associated with a cortical spreading depression wave suppress spike-wave discharges in rats. AB - To study the role of the cortex and sub-cortical structures in the generation of epileptic spike-wave discharges in more detail, cortical and striatal activity was eliminated by the induction of spreading depression in a non-invasive way. EEG and DC potentials were recorded from the cortex and striatum of WAG/Rij rats. Several of these rats show two forms of generalised epilepsy: spontaneously occurring non-convulsive absence seizures, together with convulsive audiogenic seizures. The latter can be evoked by a brief sound stimulation, provoking a fit of wild running, which is regarded as the first phase of an audiogenic seizure. In a majority of fits the cortical DC potential does not show main changes, while the spontaneously occurring spike-wave discharges are briefly suppressed for some minutes. In a minority of fits, however, audiogenic seizures are associated with a spreading depression wave, clearly expressed in the cortical DC potential. This wave is bilaterally initiated in the cortex and propagates to the caudate nucleus of the striatum. In these cases spontaneously occurring spike-wave discharges are fully suppressed for about 1 h. It is suggested that cortical spreading depression, triggered by a short audiogenic seizure, induces a long-lasting suppression of spike-wave discharges. These results are in line with the concept that spike-wave discharges are originally initiated in the cortex, as proposed by the 'cortical focus' theory. The precise role of the striatum remains less clear, although this structure seems not to play a pivotal role in spike-wave generation. PMID- 16212993 TI - Losartan blocks drinking and cFos expression induced by central ornithine vasotocin in rats. AB - We previously reported that an intracerebroventricular (icv) injection of the oxytocin receptor antagonist ornithine vasotocin (OVT) caused water and saline intakes, a pressor response, and Fos-like immunoreactivity (Fos-IR) in the median preoptic nucleus of the rat brain. In the present report, rats receiving an icv injection of isotonic saline vehicle followed by an icv injection of 10 microg of OVT 20 min later drank 5.5+/-1.1 ml of total water and saline intake in 60 min after the OVT; rats receiving 10 microg of losartan before the OVT drank only 0.9+/-0.3 ml of total fluid. In a separate study, rats were treated as above except that they were not allowed to drink and were perfused for analysis of Fos IR in the median preoptic nucleus at 90 min. Fos-IR in the dorsal part of the median preoptic nucleus was significantly suppressed from 2.69+/-0.57 cells per 10,000 square mum in vehicle-treated rats to 0.89+/-0.20 in losartan-treated rats. Losartan alone had no effect on Fos-IR. Losartan did not reduce intake of saccharin in a dessert test. This suggests that the OVT-induced drinking may result from an activation or disinhibition of angiotensin type AT1 receptors in the median preoptic nucleus. PMID- 16212995 TI - Meeting the information needs of the 'Health for all' challenge--lessons from Scotland. AB - OBJECTIVE: To create a public health data resource for Scotland that is based on a socio-ecological model of the determinants of health and would, therefore, meets the needs of the emerging public health agenda. DESIGN: Action research, in which the approach moved logically through stages of action (conception, feasibility study, pilot projects, leading to a national set of integrated health and well-being profiles). Each stage built on the results of the previous research. RESULTS: The conceptual stage identified the need for an approach to public health data that kept pace with the increasingly accepted socio-ecological models of the determinants of health. A feasibility study concluded that sufficient data were available to populate the health fields that represented the important determinants of health. At this time strengths and weakness in data were defined. This led to the articulation of a 'vision' for integrated public heath data in Scotland that was the subject of a wide consultation. Pilot studies provided local stakeholders with imaginatively presented data (on population demographics, health and function, behaviour, social environment, economy, physical environment, morbidity and mortality) for their local communities. The response to these was so positive that a demand was created for a comprehensive set of 'community profiles'. These, in addition to parliamentary constituency profiles, have now been created and widely disseminated. CONCLUSIONS: It has been possible, despite many difficulties, to develop approaches to public health information that are informed by the socio-ecological model of health and create outputs that represent a significant advance on previous approaches to public health data. This is a work in progress and many issues remain unresolved. Interaction with others engaged in parallel tasks will facilitate the next steps. PMID- 16212996 TI - The performance of RT-PCR compared with a rapid serological assay for acute dengue fever in a diagnostic laboratory. AB - The laboratory diagnosis of dengue has largely relied on serological assays, although many different RT-PCR protocols have been reported. Owing to its limited use, the value of RT-PCR in the clinical laboratory has not been fully evaluated. During the outbreak of severe acute respiratory syndrome (SARS) in Singapore in 2003, RT-PCR to detect dengue viral RNA was used as a rapid diagnostic tool to differentiate dengue from SARS among patients who presented to a hospital designated to manage and quarantine SARS cases. A total of 343 results for RT-PCR and 439 results for serology were analysed and compared with the final discharge diagnosis. Our experience indicates that RT-PCR for dengue can be set up rapidly in a clinical laboratory, with very sensitive and specific results for the diagnosis of dengue, particularly in the first 5 days from onset of symptoms. PMID- 16212997 TI - Decision-making in variable environments-- a case of group selection and inter generational conflict? AB - Suppose organisms need to engage in a particular action exactly once during some fixed period of time. Further suppose they can time this action to optimise their fitness based on the expected current payoff and the probability distribution of later payoffs. For an example we consider the timing of the annual nuptial flight in eusocial insects. Using two population genetics models, we ask whether stochasticity leads to evolutionary conflict between the queen and her offspring. We find that the winning phenotype is independent of who controls the timing. The best response to any non-equilibrium population strategy is the same in both control scenarios, a result that carries over to the diploid case. Although inter generational conflict is therefore ruled out, the models support a previous observation that at equilibrium some of the offspring have a lower expected payoff than others. By measuring fitness in terms of relative reproductive success, we show that all individuals are in fact equally well off making group selectionist arguments unnecessary. As such, the models should improve our understanding of the difficult conceptual problem of the unit of natural selection in stochastic environments. PMID- 16212998 TI - Pair statistics clarify percolation properties of spatially explicit simulations. AB - Dispersal is a fundamental control on the spatial structure of a population. We investigate the precise mechanism by which a mixed strategy of short- and long distance dispersal affects spatial patterning. Using techniques from pair approximation and percolation theory, we demonstrate that dispersal controls the extent to which a population is completely connected by modulating the proportion of neighboring sites which are simultaneously occupied. We show that near the percolation threshold this pair statistic, rather than other metrics proposed earlier, best explains clustering, and we suggest more general circumstances under which this may hold. PMID- 16212999 TI - Effects of interchanging hyperopic defocus and form deprivation stimuli in normal and optic nerve-sectioned chicks. AB - To test the hypothesis that the same mechanisms mediate form deprivation and lens induced myopia, the ocular growth responses of chicks alternately exposed to lenses and diffusers at regular intervals (3h) were compared to those of chicks exposed to either negative lenses or diffusers alone. In total, there were four experiments: (1) -15 D lenses and/or diffusers on normal birds, (2) -15 D lenses and/or diffusers on optic nerve-sectioned (ONS) birds, (3) -5/-10/-15 D lenses (sequentially applied) and/or diffusers on normal birds and (4) -5/-10/-15 D lenses and/or diffusers on ONS birds. All treatments were monocular. In all experiments, optical axial lengths (cornea-to-retina distances) in treated eyes were greater than in fellow eyes, irrespective of the optical device (diffuser, lens or switch), lens power (fixed or incremented) and optic nerve condition (intact or severed). In normal chicks, optical axial length responses in the switch group were significantly reduced relative to those of the diffuser but not to those of the -15 D lens group. For both groups of ONS birds, diffusers exaggerated the optical axial length changes. For all groups, the responses to the switch and lens groups were most similar. These results together suggest that the mechanisms mediating form deprivation- and lens-induced myopia are different. PMID- 16213001 TI - Contrast sensitivity for letter optotypes vs. gratings under conditions biased toward parvocellular and magnocellular pathways. AB - This study examined the extent to which letter optotypes and grating stimuli provide equivalent measures of contrast sensitivity under conditions designed to favor the magnocellular (MC) and parvocellular (PC) pathways. The contrast sensitivity functions (CSFs) of three visually normal observers were measured for Sloan letters and Gabor patches, using steady- and pulsed-pedestal paradigms to bias processing toward MC and PC pathways, respectively. CSFs for Gabor patches were low-pass for the steady-pedestal paradigm and band-pass for the pulsed pedestal paradigm, in agreement with previous reports. However, CSFs for letters were low-pass for both testing paradigms. CSFs for letters restricted in frequency content by spatial filtering were equivalent to those for Gabor patches for both testing paradigms. Results indicate that conventional letter optotypes can provide a misleading measure of contrast sensitivity, especially under conditions emphasizing the PC pathway. The use of spatially band-pass filtered letters can provide a more appropriate evaluation of spatial contrast sensitivity while maintaining some of the potential advantages of letters. PMID- 16213002 TI - Classification of bathing water quality based on the parametric calculation of percentiles is unsound. AB - Analyses of Irish bathing water quality data sets are reported to investigate whether the parametric calculations proposed in the draft Bathing Water Directive are valid. Faecal coliforms (assumed to be Escherichia coli) and faecal streptococci (assumed to be intestinal enterococci) have been analysed separately. It was noted that classifications based on the parametric 95th percentile calculations disagreed with those based on percentage compliance with the standards on 13.8% of occasions. When these disagreements were studied, it was found that the datasets frequently contained many censored data points (Result < 1). Also, the datasets were not log normally distributed on at least 85% of occasions. Both these findings fatally undermine the validity of using a parametric method for calculating 95th percentiles to classify bathing water quality. By contrast the non-parametric Hazen method is a better estimate of true population 95th percentiles, but essentially gives very similar classifications to the percentage compliance approach, fully agreeing on over 95% of occasions. The same is also true when considering 90th percentiles. A series of Monte Carlo studies were also conducted to determine the impact of small numbers. It was ascertained that small sample sizes are very undependable in determining bathing water classification and the parametric method in particular is particularly unreliable. In conclusion, the parametric method for calculating bathing water compliance is so severely and fatally flawed statistically that it should not form the basis of any legislation. The Hazen method gives a better estimate of true 90th or 95th percentiles, though as the resultant classifications agree with percentage compliance so closely it is doubtful that there is any statistical value in using a percentile approach over the long established and well understood percentage compliance approach. PMID- 16213000 TI - Phosphatase enzyme histochemistry for studying vascular hierarchy, pathology, and endothelial cell dysfunction in retina and choroid. AB - Phosphatase enzymes cleave an inorganic phosphate from a substrate. Phosphatase enzyme histochemistry followed by flat-embedding in glycol methacrylate is extremely useful in studying retinal and choroidal vascular development and loss, since only viable blood vessels have these enzyme activities. Sites of occlusion and remodeling can be identified and analysed, resulting in new insights into the cause of occlusion. The phosphatase activities are elevated in neovascularization making possible high resolution analysis of neovascularization, the feeder vessels, and the retinal milieu in which angiogenesis occurs. Adenosine diphosphatase (ADPase) catalyzes ADP to an inorganic phosphate plus adenosine monophosphate, preventing accumulation of ADP, one of the most potent stimuli for platelet aggregation. The ADPase technique can be used in any species but this report highlights its use in dog and human retinas. The ADPase technique has yielded important insights into vaso-occlusive and vasoproliferative processes in retinopathy of prematurity, sickle cell and diabetic retinopathies. The alkaline phosphatase flatembedding technique is useful in evaluating dog, cat, and human choroidal vasculatures. It has permitted quantification of the loss of choriocapillaris in diabetic choroidopathy and of the RPE and choriocapillaris in geographic atrophy and exudative age-related macular degeneration. PMID- 16213003 TI - Respirometric calibration and validation of a biological nitrite oxidation model including biomass growth and substrate inhibition. AB - The modelling of the nitrification process of high-strength ammonium wastewater must be designed to consider it as a two-step reaction with substrate inhibition. Consequently, kinetic and stoichiometric parameters of both steps are required. In this work, the second step in the nitrification process was studied: a biological nitrite oxidation model was formulated, calibrated and validated using only oxygen uptake rate (OUR) measurements. The model included biomass growth and substrate inhibition. First, the biomass yield coefficient for nitrite-oxidising biomass was determined. Then, a respirometric experiment with one nitrite pulse of 500 mg N-NO2- L(-1) was performed to estimate the rest of the model parameters. The practical identifiability study showed that the parameters were strongly correlated. Hence, a new experimental design consisting of two consecutive pulses and a delayed third one was designed to improve the parameter identifiability. Both experimental designs were compared using contour plots of the objective function and optimal experimental design criteria for parameter estimation. It was concluded that the parameter identifiability was improved with the new experimental design. Finally, the estimated parameters were validated and the pH effect on the inhibition coefficient was evaluated. PMID- 16213004 TI - Microbial community activities during establishment, performance, and decline of bench-scale passive treatment systems for mine drainage. AB - Permeable reactive barrier (PRB) technology, in which sulfate-reducing bacteria (SRB) facilitate precipitation of metal sulfides, is a promising approach for remediation of sulfate- and metal-laden mine drainage. While PRBs are easily established, they often decline for reasons not well understood. SRB depend on or compete with multiple dynamic microbial populations within a PRB; as a result, performance depends on the changing PRB chemical composition and on succession and competition within the microbial community. To investigate these interactions, we constructed and monitored eight bench-scale PRBs to define periods of establishment, performance, and decline. We then conducted short-term batch studies, using substrate-supplemented column materials, on Days 0 (pre establishment), 27 (establishment), 41 (performance), and 99 (decline) to reveal potential activities of cellulolytic bacteria, fermenters + anaerobic respirers, SRB, and methanogens. PRBs showed active sulfate reduction, with sulfate removal rates (SRR) of approximately 1-3 mol/m3/d, as well as effective removal of Zn2+. Potential activities of fermentative + anaerobic respiratory bacteria were initially high but diminished greatly during establishment and dropped further during performance and decline. In contrast, potential SRB activity rose during establishment, peaked during performance, and diminished as performance declined. Potential methanogen activity was low; in addition, SRB-methanogen substrate competition was shown not to limit SRB activity. Cellulolytic bacteria showed no substrate limitation at any time. However, fermenters experienced substrate limitation by Day 0, SRB by Day 27, and methanogens by Day 41, showing the dependence of each group on upstream populations to provide substrates. All potential activities, except methanogenesis, were ultimately limited by cellulose hydrolysis; in addition, all potential activities except methanogenesis declined substantially by Day 99, showing that long-term substrate deprivation strongly diminished the intrinsic capacity of the PRB community to perform. PMID- 16213005 TI - Phytoplankton size-distribution and community structure in relation to mucilage occurrence in the northern Adriatic Sea. AB - The spatial and temporal variations of pico-, nano- and microphytoplankton abundance and composition were investigated over a 37 month period, focusing on the ecological role of different size classes of phytoplankton, and on the changes of the community structure that might occur during periods when large mucilage macroaggregates appear. Samples were collected monthly from June 1999 to July 2002 at 11 stations, along three transects covering the northern Adriatic basin. Highest abundances were observed in late-winter/spring for microphytoplankton (mainly diatoms), in spring-summer for nanophytoplankton, and in summer for picophytoplankton. The autotrophic component was more abundant in the summers of 2000 and 2002 (when large mucilage aggregates occurred) than in the summers of 1999 and 2001 (when a massive phenomenon was not observed). This increase was statistically significant for pico-, nano- and, among microphytoplankton, only for dinoflagellates. Blooms of picophytoplankton were often observed at the bottom layer during mucilage summers. The microphytoplankton community during mucilage phenomena was characterized by a species composition (Chaetoceros spp., Cerataulina pelagica, Pseudo-nitzschia delicatissima, P. pseudodelicatissima, Cylindrotheca closterium, Dactyliosolen fragilissimus) comparable to that observed in summers without extensive mucilage occurrence. However, some species appeared with significantly higher densities in the summers of 2000 and 2002: Ceratium furca, C. closterium, Oxytoxum spp., Hemiaulus hauckii and Gonyaulax fragilis. Microscopic observation of aggregates revealed that the microphytoplankton species composition inside the aggregates was comparable to that observed in the water column, with an enrichment of opportunistic species such as C. closterium and P. delicatissima. The presence of mucilage aggregates affects the phytoplankton populations in the water column, even when aggregates are at early stages. It seems that there is a mutual relationship between phytoplankton and aggregates, i.e., several diatom and dinoflagellate species may contribute to the aggregate formation and enlargement, but mucilage aggregates themselves may also affect the phytoplankton populations, allowing the development of a rich diatom community and in general enhancing nanophytoplankton growth. PMID- 16213006 TI - Evaluation of DGT as a metal speciation tool in wastewater. AB - This paper aims to evaluate the performance of the diffusive gradient in thin film technique (DGT) as a speciation tool for metals in wastewater. The validity of metal sampling by DGT in wastewater was checked. DGT was used in parallel with the Daphnia magna acute toxicity test in order to obtain information on the speciation of copper and cadmium in diluted and spiked filtered wastewater (raw and treated) from two treatment plants. Combining the chemical (DGT) and the biological methods (D. magna toxicity test) allowed metal to be fractionated into inorganic, labile organic and inert organic metal. Copper was mainly found as inert organic complexes, whereas the major part of cadmium was found to be labile organic complexes. The proportion of inert organic copper complexes was higher in the presence of treated wastewater than in raw wastewater. The use of restricted gels in DGT devices discriminated more labile organic cadmium than labile organic copper, indicating that cadmium weak ligands have more complex structures than copper weak ligands. In our experimental conditions (i.e. a high metal to ligand ratio), DGT, even equipped with restricted gels, was able to accumulate labile organic complexes. This result highlights that the ecotoxicological interpretation of DGT measurement should be considered carefully. DGT is a reliable tool to assess the chemical characteristics of metals (i.e. reactivity) in wastewater, but it does not ensure that only inorganic metal is measured. PMID- 16213008 TI - Long-term survival after concomitant chemoradiotherapy prior to surgery in advanced cervical carcinoma. AB - OBJECTIVE: To report the long-term survival of 35 patients who underwent surgery after concomitant chemoradiation for locally advanced cervical cancers. METHODS: From 1988 to 1992, 20 bulky IB-IIB patients and 15 stage III-IVA patients underwent surgery after concurrent chemotherapy (CDDP and 5-FU) and radiotherapy. 26 had a hysterectomy, 8 had an exenteration, 1 had no tumor resection. 21 had a para-aortic lymphadenectomy. Endpoints were recurrence and distant metastasis rates, overall survival (OS) and disease-free survival (DFS) at 5 and 10 years. Analysis included FIGO stage, type of surgery (palliative or curative), response to chemoradiation or para-aortic lymphatic status. RESULTS: Surgery had been only palliative in 6 cases (17.1%). A pelvic control has been achieved in 31 patients (88.6%). Pelvic recurrences occurred after a median interval of 7 months. Distant metastases occurred in 10 patients (28.6%), after a median interval of 20 months. So far, 16 patients have died (45.7%). The 10-year DFS is 56.7% in the whole series. Only the type of surgery significantly affected the OS. Only the para aortic lymphatic status significantly affected the DFS. CONCLUSION: Associating chemoradiation with curative surgery, we obtained a 10-year DFS of 66.4% (OS 57.7%). Adjuvant surgery may increase the survival as it reduces the risk of local relapse. PMID- 16213007 TI - Adjuvant whole abdominal irradiation in clinical stages I and II papillary serous or clear cell carcinoma of the endometrium: a phase II study of the Gynecologic Oncology Group. AB - OBJECTIVES: To evaluate outcome in patients with clinical stage I/II papillary serous (PS) or clear cell (CC) endometrial carcinoma treated with whole abdominal radiotherapy. METHODS: After total abdominal hysterectomy with bilateral salpingo oophorectomy, pelvic/para-aortic lymph node sampling, and peritoneal washings, eligible patients received radiotherapy (RT) to the abdomen (3000 cGy at 150 cGy/day) with a pelvic boost (1980 cGy at 180 cGy/day). RESULTS: Among 21 PS patients (median age: 68 years), one refused therapy, and another received a non protocol vaginal boost. In total, eight patients died of disease (DOD) between 9.6 and 35.2 months. Five others died due to protocol treatment (1), toxicity from subsequent chemotherapy (1), intercurrent disease (1), and unknown cause (2). Five-year progression-free survival (PFS) was 38%. Among treated patients who DOD, sites of recurrence included lung (2), lung/vagina (1), abdomen/pelvis (1), vagina (1), and abdomen (2). Among 13 CC patients (median age: 63 years), one received pelvic RT only and died with intercurrent disease. Five others died due to DOD (3), intercurrent disease (1), and unknown cause (1). Five-year PFS was 54%. Among patients who DOD, sites of recurrence included lung (1), vagina (1), and unknown (1). Grade 3/4 toxicities for both histologic groups included gastrointestinal (three grade 4; three grade 3), hematologic (one grade 4), and cutaneous (one grade 3). CONCLUSIONS: Over half of the treatment failures were within the radiation field. Systemic chemotherapy, radiosensitizing chemotherapy, or sequential radiation and chemotherapy should be considered in future adjuvant trials for these patients. PMID- 16213009 TI - Human papillomavirus vaccines in development: if they're successful in clinical trials, how will they be implemented? AB - INTRODUCTION: There are some 30-40 genotypes of human papillomavirus (HPV) that cause anogenital lesions in humans, primarily cervical, vaginal, and vulvar dysplasias and cancer in women, anal dysplasias in some men, and genital warts in both sexes. METHODS: Both GlaxoSmithKline and Merck companies are developing vaccines for the prevention of HPV infection. RESULTS: Both vaccines stimulate an antibody response against HPV types 16 and 18; the Merck vaccine also include HPV types 6 and 11. CONCLUSIONS: This report focuses on progress with both prophylactic vaccines and introduces strategies for global immunization. PMID- 16213010 TI - MRP2 (ABCC2) and cisplatin sensitivity in hepatocytes and human ovarian carcinoma. AB - OBJECTIVE: The ABC transporter MRP2 (ABCC2) can mediate cisplatin efflux, and over-expression of MRP2 has been associated with cisplatin resistance in cancer cell lines. The aim of this study was to determine the role of MRP2 in modulating cisplatin cytotoxicity in normal cells as well as the relationship between MRP2 expression and clinical response to platinum-based agents in ovarian cancer. METHODS: The effect of absence of MRP2 expression on cisplatin sensitivity was investigated using primary hepatocyte cultures from the TR- rat strain, which is deficient in Mrp2. We also examined MRP2 expression immunohistochemically in human ovarian tumors exhibiting extremes of clinical response to platinum-based chemotherapy, either absolute platin resistance or patients with residual disease after surgery who experienced extremely long complete response to primary platinum-based chemotherapy. RESULTS: Primary hepatocyte cultures from Mrp2 deficient TR- rats were over threefold more sensitive to cisplatin and accumulated a twofold greater amount of platinum on DNA that wild-type rat hepatocytes. In human ovarian carcinomas, MRP2 was detected by immunohistochemistry in 3/13 (23%) tumors from patients with absolute platin resistance compared with 5/9 (56%) tumors from patients with prolonged survival following treatment including a platinum-based agent. CONCLUSION: These studies indicate that MRP2 may play an important role in modulating normal tissue response to cisplatin. However, MRP2 expression occurred only in a subset of primary ovarian cancers, was frequently aberrant in location and was not correlated with clinical response to platinum-based chemotherapy. PMID- 16213011 TI - Carcinosarcoma of the ovary-a case series. AB - OBJECTIVE: To evaluate our experience with ovarian carcinosarcoma and identify prognostic factors. METHODS: Thirty-one cases of ovarian carcinosarcoma were identified over a 6-year time period through tumor registry and pathology records. Fisher exact test and log rank using Kaplan-Meier method (P < 0.05) were used to compare variables with outcome. RESULTS: All 31 patients underwent initial surgical treatment with an appropriate staging procedure. Stage distribution: 1 stage I, 6 stage II, 23 stage III, and 1 stage IV. The median follow-up was 28 months. The median survival for the entire group was 21 months. Early vs. advanced stage significantly influenced progression-free interval, P = 0.05. Nineteen patients were found to have stage IIIC disease and required debulking procedures. In patients with stage IIIC disease, presence of residual disease was associated with decreased overall survival, P = 0.03. 29 patients received adjuvant chemotherapy with 11 patients receiving ifosfamide/cisplatin and 16 patients receiving carboplatin/taxol. Progression-free interval was improved with the use of ifosfamide/cisplatin vs. carboplatin/taxol. The median PFI was 12 months in the carbo/taxol group and has not been reached in the ifos/cisplatin group (P = 0.005). The overall survival was also significantly improved with the use of ifosfamide/cisplatin, P = 0.03. In advanced stage patients, overall survival was not significantly influenced by type of adjuvant chemotherapy administered, P = 0.13. CONCLUSIONS: Ovarian carcinosarcoma has a poor overall prognosis with median survival rates reported in the literature ranging from 7-10 months. Our series, although limited by a small number of patients, exhibits a more encouraging median survival of 21 months for the overall group. Aggressive debulking to eliminate residual disease and the use of ifosfamide/cisplatin chemotherapy seem to be factors in this improved outcome. PMID- 16213012 TI - The effect of long non-reproductive periods on the genital health in captive female white rhinoceroses (Ceratotherium simum simum, C.s. cottoni). AB - White rhinoceroses suffer from a low reproductive rate in captivity. Intensive efforts to propagate specifically the northern white rhinoceros have been very limited. The dismal outlook for this subspecies in the wild makes successful ex situ breeding programs paramount. In this context, this study examined 48 southern and 6 northern white rhinoceroses using ultrasound and faecal hormone analysis to elucidate causes for female reproductive failure and to determine whether long non-reproductive periods have a detrimental impact on genital health. Results showed that 76% of the nulliparous females had intact hymenal membrane indicating these females had never been bred, at an age when their wild counterparts have delivered multiple offspring. Fifty-six percent of the studied population had various reproductive pathology. Cystic endometrial hyperplasia; leiomyomas of the cervix, uterus and ovary, adenoma; para-ovarian cysts and hydromucometra represent the scope of lesions identified. The stages of the lesions in nulliparous females correlated with age (r = 0.4, P < 0.05). Due to the severity of the lesions, 28% of the study population was considered post reproductive. Therefore, the reproductive life span in some individuals was 10-20 years shorter than expected. However, in parous females the incidence of pathological lesions was significantly lower (P < 0.0001). Seventy-eight percent females studied had erratic or absent luteal activity. The hormone data corresponded with two ultrasonographic levels of ovarian activity, active and inactive, occurring within an age range of 3-19 years and 15-38 years, respectively. This suggests the lack of ovarian activity by reproductive mid-life in non-reproducing females. The accuracy of the ultrasound findings was validated by necropsy in nine animals showing a strong positive correlation (r2 = 0.9, p < 0.001). Our data suggests that the development of reproductive pathology and ovarian inactivity in white rhinoceros is an age-related consequence of long non reproductive periods. This asymmetric ageing process of the genital organs can be prevented with the achievement of at least one pregnancy. PMID- 16213013 TI - Changes in uterine protein secretion during luteal and follicular phases and detection of phosphatases during luteal phase of estrous cycle in buffaloes (Bubalus bubalis). AB - Changes in uterine proteins during different reproductive states and their functional significance though known in other species have not been established in buffaloes. An attempt has been made to unravel the changes in composition of buffalo uterine secretion with growth and regression of corpora-lutea during early, mid and late luteal and follicular phase of estrous cycle using gel filtration and electrophoresis techniques. Also the phosphatases activities in luteal phase uterine secretions have been studied. Gel filtration chromatography analysis revealed a protein peak in void volume of the column, the intensity of which was more in all the luteal phase samples than follicular phase samples. Alkaline phosphatase was also found eluted in the void volume. The other three uterus-specific peaks (Peaks V-VII) were detected below 13.7 kd molecular weight. There were at least five peaks of acid phosphatases activity in chromatogram. Silver staining of SDS-PAGE gel detected as many as 40 protein bands in the uterine fluid of which nine proteins were glycoproteins. Molecular weight (MW) comparison revealed the major protein band at 66 kd which could be serum albumin. Comparison of uterine proteins with serum protein bands revealed a 93.5 kd glycoprotein in buffalo serum that did not appear in uterine fluid and at least 11 uterus-specific protein bands (506, 470, 241, 114, 49, 38, 33, 26, 19.2, 16, and 14.3 kd). The 38 and 19.2 kd bands were luteal-stage specific. Intense periodic acid Schiff's (PAS) stained bands in uterine proteins compared to serum indicated glycosylation process in endometrial epithelial cells. The study suggested that buffalo uterine secretion contained mainly serum and several uterus-specific proteins of which few were luteal phase specific. Further study on characterizing the unique or most abundant proteins and defining their role in uterine functions would help to address the cause of low reproduction rate in buffaloes. PMID- 16213014 TI - Soluble osteogenic molecular signals and the induction of bone formation. AB - The induction of bone formation starts by erecting scaffolds of smart biomimetic matrices acting as insoluble signals affecting the release of soluble osteogenic molecular signals. The cascade of bone differentiation by induction develops as a mosaic structure singly initiated by the osteogenic proteins of the transforming growth factor-beta (TGF-beta) supergene family. The osteogenic signals when combined with an insoluble signal or substratum initiate de novo bone formation by induction and are deployed singly, synergistically and synchronously to sculpt the architecture of the mineralized bone/bone marrow organ. The osteogenic proteins of the TGF-beta superfamily are the common molecular initiators deployed for embryonic development and the induction of bone in postnatal osteogenesis, whereby molecules exploited in embryonic development are re-deployed in postnatal tissue morphogenesis as a recapitulation of embryonic development. The pleiotropy of the osteogenic proteins of the TGF-beta superfamily is highlighted by the apparent redundancy of molecular signals initiating bone formation by induction including the TGF-beta isoforms per se, powerful inducers of endochondral bone but in the primate only. Bone induction by the TGF-beta isoforms in the primate is site and tissue specific with substantial endochondral bone induction in heterotopic sites but with absent osteoinductivity in orthotopic calvarial sites on day 30 and only limited osteogenesis pericranially on day 90. Ebaf/Lefty-A, a novel member of the TGF-beta superfamily, induces chondrogenesis in calvarial defects of Papio ursinus and bone regeneration across the defect on day 30 and 90, respectively. The strikingly pleiotropic effects of the bone morphogenetic and osteogenic proteins (BMPs/OPs) spring from amino acid sequence variations in the carboxy-terminal domain and in the transduction of distinct signalling pathways by individual Smad proteins after transmembrane serine/threonine kinase complexes of type I and II receptors. Predictable bone regeneration in clinical contexts requires information concerning the expression and cross regulation of gene products of the TGF-beta superfamily. OP-1, BMP-3, TGF-beta1 and type IV collagen mRNAs expression correlates to the morphological induction and maintenance of engineered ossicles by the hOP-1 osteogenic devices in the non human primate P. ursinus. Amino-acid sequence variations amongst BMPs/OPs in the carboxy terminal domain confer the structure/activity profile responsible for the pleiotropic activity that controls tissue induction and morphogenesis of a variety of tissues and organs by different BMPs/OPs which are helping to engineer skeletal tissue regeneration in molecular terms. PMID- 16213016 TI - Transcriptional and post-transcriptional regulation of glutathione S-transferase P1 expression during butyric acid-induced differentiation of K562 cells. AB - Over-expression of glutathione S-transferase P1 is related to chemotherapeutic drug resistance as well as to differentiation of human erythroleukemia cells. In opposition to previously described differentiating inducers which enhance the GST resistance phenotype, time- and concentration-dependent activation of both erythroid and megakaryocytic differentiation pathways by butyric acid progressively diminished GSTP1 mRNA expression. GSTP1 mRNA expression decreased by 25% (p<0.01) and 64% (p<0.01) in 1mM and 2mM butyric acid-differentiated K562 cells, respectively. These results were associated to both a reduction of GATA-1 binding activity to the GSTP1 promoter and to a posttranscriptional destabilization of GSTP1 mRNA in a concentration dependent manner. Indeed, GSTP1 mRNA half-life decreased from 43.8 to 36.2 h and 12.6 h in 1mM- and 2mM-treated cells, respectively. PMID- 16213015 TI - Blockade of angiotensin II AT1 receptors inhibits pressor action of centrally administered interleukin-1beta in Sprague Dawley rats. AB - The aim of the present study was to evaluate the influence of intraventricular administration of recombinant rat interleukin-1beta (IL-1beta) on regulation of resting blood pressure and heart rate and to test the hypothesis that the brain angiotensinergic system is involved in regulation of hemodynamic parameters by centrally applied IL-1beta. The experiments were performed on Sprague Dawley rats, assigned to three series of experiments. In series 1 (control, n = 6), mean arterial pressure (MAP) and heart rate (HR) were recorded for 15 min under baseline conditions. This was followed by infusion of saline (0.9% sterile NaCl 5 microL/h) into the left cerebral ventricle (LCV). Measurements were continued during the next 60 min. In series 2 (n = 6) and 3 (n = 6) the experimental design was similar, except that in series 2 the animals were LCV infused with saline containing IL-1beta (100 ng/h) and in series 3 with saline containing IL-1beta (100 ng/h) and angiotensin type 1 (AT1) receptors antagonist (Losartan, 10 microg/h). LCV infusion of saline alone did not influence MAP and HR while administration of IL-1beta elicited significant increase in MAP, but not in HR. The pressor effect was absent during combined infusion of IL-1beta and Losartan. Results of the study provide evidence that centrally administered IL-1beta exerts pressor effect, and reveal that this effect is mediated by stimulation of the brain angiotensin system and requires activation of AT1 receptors. PMID- 16213017 TI - Interspecific variation in heavy metal body concentrations in biota of Sunderban mangrove wetland, northeast India. AB - The coastal environment of West Bengal, recognized as the most diversified and productive ecosystem among all the maritime states of India, faces organic pollution from domestic sewage and urban and industrial effluents leading serious impacts on biota. The present paper aims at providing information on concentration level of heavy metals among the tissues of benthic polychaetes, bivalve molluscs and finfishes collected from Sundarban mangrove wetland, northeast coast of India. An overall common trend in bioaccumulation was revealed with the following decreasing order: Zn>Mn>Cu>Cr>Se>Hg with few exceptions. Both species dependent variability and temporal variations were pronounced. A high degree of organ specificity was evident in the bivalves where gill and mantle exhibited higher metal accumulation due to ion exchange property of the mucous layer covering these organs. Variability between closely related species is a reflection of different uptake rates, physiology and impact of environmental factors. The results of this analysis suggest that mollusks can play a significant role in trace metal trophic transfer studies, especially as their representatives are intertidally and subtidally ubiquitous. PMID- 16213018 TI - Distribution and fate of persistent organochlorine pesticides in coastal marine environment of Mumbai. AB - Multi-compartment monitoring of residue levels of OCPs in coastal marine environment of Mumbai has been studied. The concentration of total HCHs in seawater varied from 0.16 to 15.92 ng/L and concentrations of total DDT varied from 3.01 to 33.21 ng/L. The total HCH concentration in the sediment samples was in the range of 3.8 to 16.2 ng/g. Gamma-HCH contributed almost 55% to the total HCH. In sediment samples the DDT has higher mean concentration in comparison to its metabolite DDE and DDD. The concentration of total HCHs in different marine species varied from 0.87 to 33.73 ng/g and concentrations of total DDT varied from 0.38 to 34.1 ng/g. The variation in the beta-HCH in different compartments is not significant and this could be due to the high persistence and metabolically inactive nature of this isomer. The alpha-HCH is found to be more dominant in fish samples whereas the gamma-HCH is major contributor in the sediment samples. PMID- 16213019 TI - Comparing environmental issues in Cuba before and after the Special Period: balancing sustainable development and survival. AB - Following the Earth Summit in 1992, Cuba designed and implemented a variety of programs, administrative structures, and public awareness activities to promote sound environmental management and sustainable development. This came shortly after the fall of the Soviet Union and the strengthening of the US blockade in 1990, which resulted in a 35% drop in Cuban GDP. This period, referred to as the Special Period, witnessed a decrease in many environmentally damaging activities both by choice and by necessity, but also resulted in many decisions to resuscitate the Cuban economy. The purpose of this work was to compare and rank the environmental risks Cuba faced before and during the Special Period (1990 2000) using two Comparative environmental risk assessments (CERAs). To do so, an ecosystem integrity risk assessment matrix was constructed with 42 risk end points. The matrix assessed the risk posed by 17 problem areas including air pollution, water contamination, solid waste sites, pesticides and ecosystem degradation. The risks were calculated using five criteria: area affected, vulnerability of affected population, severity of impact, irreversibility of effect and uncertainty. To construct this matrix, both literature reviews and expert interviews in Cuba were conducted in 2000. The results showed a general decrease in risk scores during the Special Period. Before the Special Period, high risks were posed by: terrestrial degradation and industrial wastewater and sludge, followed by freshwater degradation, surface water stressors, and pesticides. After the Special Period, industrial wastewater and sludge and pesticides were no longer high-risk areas, but municipal wastewater and marine coastal degradation ranked higher than previously. Also, the risk endpoints most stressed after 1990 were affected by activities controlled by the government, such as mining and tourism, and lack of infrastructure. Therefore, the claims that public environmental education is the main pathway to sustainable development in Cuba seem uninformed and other management practices should be evaluated. PMID- 16213020 TI - Using nematodes in soil ecotoxicology. AB - Nematodes represent a very abundant group of soil organisms and non-parasitic species are important for soil quality and in the soil food web. In recent years, it has been shown that nematodes are appropriate bioindicators of soil condition and they are also suitable organisms for laboratory toxicity testing. The aims of this paper are to overview and critically assess methods and approaches for researching soil nematode ecotoxicology. In natural ecosystems, nematode abundance and community structure analyses were proved to be sensitive indicators of stress caused by soil pollutants and ecological disturbance. Community structure analyses may be approached from a functional or ecological point of view; species are divided into groups according to their feeding habits or alternatively the maturity index is calculated according to their ecological strategy. Many environmental factors have the potential to affect nematode community, which consequently results in high space and time variability. This variance is major handicap in field ecotoxicological studies because pollutant nematode relationships are obscured. For prospective risk assessment of chemicals, several toxicity tests with nematodes were developed and are increasingly used. Sensitivity of these tests is comparable to tests with other soil species (e.g. enchytraeids, earthworms and springtails) while tests are less demanding to space and time. Most studies have focused on metal toxicity but organic compounds are almost overlooked. Endpoints used in tests were often mortality, reproduction or movement, but more sublethal endpoints such as feeding or biomarkers have been used recently too. Although there is an increasing amount of knowledge in soil nematode ecotoxicology, there is still a lot of various issues in this topic to research. PMID- 16213021 TI - Surfactant proteins SP-A and SP-D: structure, function and receptors. AB - Surfactant proteins, SP-A and SP-D, are collagen-containing C-type (calcium dependent) lectins called collectins, which contribute significantly to surfactant homeostasis and pulmonary immunity. These highly versatile innate immune molecules are involved in a range of immune functions including viral neutralization, clearance of bacteria, fungi and apoptotic and necrotic cells, down regulation of allergic reaction and resolution of inflammation. Their basic structures include a triple-helical collagen region and a C-terminal homotrimeric lectin or carbohydrate recognition domain (CRD). The trimeric CRDs can recognize carbohydrate or charge patterns on microbes, allergens and dying cells, while the collagen region can interact with receptor molecules present on a variety of immune cells in order to initiate clearance mechanisms. Studies involving gene knock-out mice, murine models of lung hypersensitivity and infection, and functional characterization of cell surface receptors have revealed the diverse roles of SP-A and SP-D in the control of lung inflammation. A recently proposed model based on studies with the calreticulin-CD91 complex as a receptor for SP-A and SP-D has suggested an anti-inflammatory role for SP-A and SP-D in naive lungs which would help minimise the potential damage that continual low level exposure to pathogens, allergens and apoptosis can cause. However, when the lungs are overwhelmed with exogenous insults, SP-A and SP-D can assume pro-inflammatory roles in order to complement pulmonary innate and adaptive immunity. This review is an update on the structural and functional aspects of SP-A and SP-D, with emphasis on their roles in controlling pulmonary infection, allergy and inflammation. We also try to put in perspective the controversial subject of the candidate receptor molecules for SP-A and SP-D. PMID- 16213022 TI - Human peripheral blood mononuclear cells express GABAA receptor subunits. AB - The polymerase chain reaction was used to screen human peripheral blood mononuclear cells (PBMC) and Jurkat cells for the presence of GABAA receptor subunit mRNAs. Positive signals were detected for the alpha1, alpha3, beta2, beta3, delta and epsilon subunit mRNAs in both cell populations, with the Jurkat cells giving a positive signal for some additional species. Real-time PCR was used to confirm that PBMC, lymphocytes and monocytes contained significant levels of the alpha1 subunit mRNA and that PBMC and lymphocytes contained low levels of beta2 mRNA. The alpha1 subunit was detected in PBMC and fractionated T-cell populations, as well as Jurkat and HL-60 cell lines, by Western blotting and immunofluorescence using a specific antibody. The application of 1mM GABA reduced the specific increase in intracellular PBMC Ca2+ levels produced by addition of 1 nM fMLP: this effect was mimicked by muscimol, but not glycine, and was blocked by bicuculline. The inhibitory effect of GABA was limited to a subset of PBMC. We conclude that cells within the human PBMC population, including lymphocytes, express functional GABAA receptors and these receptors may modulate immune responses. PMID- 16213023 TI - A trifluoromethyl substituted organoimido derivative of the hexametalate cluster: synthesis, crystal structure and bioactivity of [Mo6O17(NAr)2]2- (Ar=o-CF3C6H4). AB - The hexamolybdate cluster has been functionalized with fluorine-containing aromatic amine by the DCC (N,N'-dicyclohexylcarbodiimide) dehydrating protocol and a novel bifunctionalized arylimido derivative of hexamolybdate bearing the electron-withdrawing trifluoromethyl group, (Bu(4)N)(2)[Mo(6)O(17)(NAr)(2)] (Ar=o CF(3)C(6)H(4)), has been prepared. Complete assignments were achieved for the title compound by elemental analysis, IR, (1)H NMR, UV/visible and single-crystal X-ray diffraction analyses. The preliminary agricultural biological activity test indicated that it has some herbicidal and insecticidal activities. PMID- 16213024 TI - The mechanism of Compound I formation revisited. AB - The most recently proposed mechanisms for the formation of the Compound I intermediates of the peroxidases and catalases have been based on the crystallographic elucidation of the enzyme structures. It has been assumed that these mechanisms are compatible with an earlier proposal of the formation of a reversible enzyme-substrate intermediate called Compound 0, which was based on data that pre-dated the availability of the enzyme structures. However, it is argued here that this is not the case and some modifications of the existing mechanism are proposed which reconcile the structural, kinetic and energetic data for the reactions. This paper focuses attention on horseradish peroxidase isoenzyme C and particularly on the acid-base properties of the imidazole side chain of distal histidine 42. This imidazole group has an exceptionally low pK(a) value in the resting enzyme, which is higher in Compound I and higher still in Compound II. The pK(a) value must also be greatly increased following Compound 0 formation so that the imidazole can become an effective proton acceptor. An explanation is offered in a dielectric insertion (DI) model, in which the peroxide substrate, or fragments thereof, screens the influence of the positively charged heme iron on the pK(a) value of the imidazole group. It is proposed that Compound 0 is converted to a second intermediate, Compound 0*, by intramolecular proton transfer along a pre-existing hydrogen bond, a process which reduces the energy requirements of charge separation in the deprotonation of hydrogen peroxide. PMID- 16213025 TI - Use of systemic cyclosporin A in a case of severe Toxocara uveitis. AB - The report describes the case of an adult patient presenting a severe, serologically proven, Toxocara endophthalmitis, unresponsive to the common surgical and medical approach (vitrectomy, anti-parasitic treatment followed by high-dose corticosteroids). The association of oral cyclosporin A to a lower dose of prednisone was successful in achieving a long-term control of the ocular inflammation without systemic side effects. PMID- 16213026 TI - Ofloxacin induced Sweet's syndrome in a patient with Crohn's disease. AB - Sweet's syndrome is an acute febrile neutrophilic dermatosis. This syndrome can be idiopathic, para-inflammatory, paraneoplastic, drug-induced, or pregnancy related. In this paper, a case of Sweet's syndrome associated with ofloxacin therapy in a patient with Crohn's disease is reported. PMID- 16213027 TI - About being BOLD. AB - The last decade has seen an unprecedented increase in the use of functional magnetic resonance imaging (fMRI) to understand the neural basis of cognition and behavior. Being non-invasive and relatively easy to use, most studies relied on changes in the blood oxygenation level dependent (BOLD) contrast as an indirect marker of variations in brain activity. However, the fact that BOLD fMRI is dependent on the blood flow response that follows neural activity and does not measure neural activity per se is seen as an inherent cause for concern while interpreting data from these studies. In order to characterize the BOLD signal correctly, it is imperative that we have a better understanding of neural events that lead to the BOLD response. A review of recent studies that addressed several aspects of BOLD fMRI including events at the level of the synapse, the nature of the neurovascular coupling, and some parameters of the BOLD signal is provided. This is intended to serve as background information for the interpretation of fMRI data in normal subjects and in patients with compromised neurovascular coupling. One of the aims is also to encourage researchers to interpret the results of functional imaging studies in light of the dynamic interactions between different brain regions, something that often is neglected. PMID- 16213028 TI - Dopamine transporter availability in medication free and in bupropion treated depression: a 99mTc-TRODAT-1 SPECT study. AB - BACKGROUND: Bupropion is thought to exert its antidepressive effect by blocking the dopamine transporter (DAT). The purpose of this study was to evaluate the DAT activity in depressed patients by means of 99mTc-TRODAT-1 SPECT in relation to the efficacy of bupropion treatment. METHODS: In 12 healthy controls and 16 depressed patients the baseline DAT activity was examined. Nine of the 16 patients went through an additional second SPECT investigation, after 4 weeks of bupropion treatment. RESULTS: In the depressed patients, the baseline DAT striatum-occipital ratio (SOR) (1.04+/-.36, mean+/-SD) was not significantly different from that in the control group (1.12+/-.33) (p>.05). Correlation was found between baseline SOR and HAM-D score change (r=-.745, p=.02) of the bupropion treated patients. The average DAT occupancy due to the bupropion treatment was 20.84+/-27.7%. No significant correlation between the therapeutical effectiveness and the occupancy was observed. LIMITATIONS: One of the limiting factors of our study has been the lack of drug monitoring. CONCLUSIONS: In good agreement with other PET studies, we found 20.84% DAT occupancy during bupropion treatment. The lack of correlation between the efficacy of therapy and occupancy of DAT may raise the question as to whether other mechanisms are involved in the effect of bupropion. PMID- 16213029 TI - Impact of childhood abuse on the course of bipolar disorder: a replication study in U.S. veterans. AB - BACKGROUND: The association between early childhood abuse and the course of illness, including psychiatric comorbidities, in adults with bipolar disorder has not been examined in a predominantly male or veteran population. METHODS: As part of the VA Cooperative Study 430, "Reducing the Efficacy-Effectiveness Gap in Bipolar Disorder," 330 veterans (91% male) with bipolar I or II disorder who were enrolled in a 3-year prospective study were examined for baseline data obtained at study entry. Diagnoses were determined by the use of the SCID. A semistructured interview designed to elicit data about exposure to childhood physical, sexual, or combined abuse was conducted as part of baseline demographic and clinical information. Other reports from this data set have not addressed the issues of childhood adversity. RESULTS: Childhood abuse was reported by 48.3% of the subjects (47.3% of men). Any abuse (AA) was reported by 48.3%; sexual abuse without physical abuse (SA) was reported by 8%, physical abuse without sexual abuse (PA) by 20.7%, and both types of abuse (combined abuse, CA) by 18.7% of the male subjects. Female veterans reported more SA (27%) and less PA (6.7%). AA subjects were more likely to have current PTSD and lifetime diagnoses of panic disorder and alcohol use disorders. CA was associated with lower SF-36 Mental scores, higher likelihood of current PTSD and lifetime diagnoses of alcohol use disorders, as well as more lifetime episodes of major depression and higher likelihood of at least one suicide attempt. Younger age at study entry was associated with AA and PA. LIMITATIONS: Potential limitations include generalizability beyond the male, veteran population of patients with bipolar disorder and the methodology used to elicit abuse histories. CONCLUSIONS: Similar to studies of predominantly female nonveteran samples, this study extends the finding that a history of childhood abuse acts as a disease course modifier in male veterans with bipolar disorder. Clinicians should routinely seek information regarding abuse and be aware that these patients may be more difficult to treat than bipolar patients who have no abuse histories. PMID- 16213030 TI - Identification and gene expression of bovine C-type lectin dectin-2. AB - The C-type lectin receptor has been shown to recognize carbohydrate moieties of self and non-self antigens, thus serving as an innate immune receptor. Using bioinformatics and molecular cloning techniques, we isolated a bovine gene that encodes a polypeptide of 206 amino acids with structural features shared by mouse and human dectin-2, including a high homology with mouse dectin-2 (66%), a type II configuration, a short cytoplasmic domain without tyrosine-based signal motifs, a carbohydrate recognition domain, a putative N-glycosylation site, and an EPN motif involved in the Ca(2+)-dependent binding of hexose carbohydrates. These results reveal this bovine gene to be a counterpart of mouse dectin-2. Moreover, the bovine dectin-2 gene showed heterogeneity in mRNA (the generation of alternatively spliced transcript) and segmentation into six exons, which are also observed in mouse dectin-2. Inconsistent with mouse dectin-2 mRNA, the bovine counterpart is abundantly expressed by Langerhans cells compared to macrophages; however, lymph nodes showed the highest expression level of bovine dectin-2, while spleen and lung showed the highest expression levels of mouse and human dectin-2. In cattle, dectin-2 expressed by dendritic cells may be clinically involved in the recognition of invading antigens in lymph nodes. PMID- 16213031 TI - Peripheral T lymphocyte changes in neonatal piglets: Relationship with growth hormone (GH), prolactin (PRL) and cortisol changes. AB - Taking into account the role played by the neuroendocrine network in affecting the early development of the immune response, the present study aims to assess neonatal immunity in piglets by testing peripheral lymphocyte age-related changes in relationship to plasma levels of some relevant immunoregulatory hormones, such as growth hormone (GH), prolactin (PRL) and cortisol. For this purpose, we studied the peripheral lymphocyte age-related changes in relationship to plasma levels of GH, PRL and cortisol in conventional piglets from birth (day 0) to 41 days of age. A significant decrease was observed in the total number of lymphocytes at day 0, with a subsequent constant increment up to 41 days of age. Concomitantly, the number of T cell subsets (mainly CD8(+) cells and double positive CD4(+)CD8(+)) was low at birth, with strong increments between the 19th and 41st days of life. The CD4(+) T cell number subset was less diminished at birth than that of CD8(+), albeit with significant increments in the post-weaning period. Of interest, gammadelta T cells, which are more involved in innate immune efficiency, displayed the same trend as CD8(+) T cells from birth to the 41st day of life. From day 0 up to the 19th day, significant inverse correlations were found between T cell subsets and GH or PRL or cortisol, albeit with more significant inverse correlations with cortisol. The high levels of GH and PRL in the pre-weaning period may be due to the fact that they have to counteract the cortisol-mediated negative effect on lymphocyte production and development. These findings suggest that stress condition occurs at birth with decreases in the immune parameters, in the same way as in human newborns, with a subsequent gradual normalisation and immune development, as shown by decreased cortisol, GH and PRL normalisation and concomitant increments in T cell subsets. PMID- 16213032 TI - Cloning and comparison of bighorn sheep CD18 with that of domestic sheep, goats, cattle, humans and mice. AB - Previously, we have shown that CD18, the beta-subunit of beta(2)-integrins, serves as a receptor for leukotoxin (Lkt) secreted by Mannheimia (Pasteurella) haemolytica on bovine leukocytes. Anti-CD18 monoclonal antibodies (mAbs) inhibit Lkt-induced cytolysis of bighorn sheep (Ovis canadensis) leukocytes suggesting that CD18 may serve as a receptor for Lkt on the leukocytes of this species as well. Confirmation of bighorn sheep CD18 as a receptor for Lkt, and elucidation of the enhanced Lkt-susceptibility of bighorn sheep polymorphonuclear leukocytes (PMNs), necessitates the cloning and sequencing of cDNA encoding bighorn sheep CD18. Hence, in this study we cloned and sequenced the cDNA encoding CD18 of bighorn sheep, and compared with that of other animal species. The cDNA of bighorn sheep CD18 has an open reading frame (ORF) of 2310bp. CD18 sequences obtained individually from peripheral blood mononuclear cells (PBMCs) and PMNs were identical to each other. Comparison of the deduced 770-amino acid sequence of CD18 of bighorn sheep with that of domestic sheep, goats, cattle, humans and mice revealed 99, 98, 95, 82 and 80% identity, respectively. Availability of cloned bighorn sheep CD18 cDNA should allow the molecular characterization of M. haemolytica Lkt-receptor interactions in bighorn sheep and other ruminants that are susceptible to this disease. PMID- 16213033 TI - Rat ultrasonic vocalization in aversively motivated situations and the role of individual differences in anxiety-related behavior. AB - Our previous work has shown that male Wistar rats, although identical in breeder, age and housing conditions, can differ systematically in their anxiety-related behavior as measured in the elevated plus-maze. Since such individual dispositions can affect the responsiveness in other aversively motivated situations, we asked in a 1st experiment whether such rats might also differ in a test of conditioned fear. Based on their levels of spontaneous open arm avoidance in the elevated plus-maze, 20 adult male Wistar rats were divided into those with "high open arm" (HOA) versus "low open arm" (LOA) time. These rats were then tested in a standard fear conditioning paradigm. During the conditioning procedure, they received 6 tone (3 kHz, 20 s) and shock (0.5 mA, 0.5 s) pairings, each followed by a 60 s inter-stimulus interval. Conditioned responses to the tone were tested 24 h thereafter. During both days, freezing behavior and ultrasonic vocalization were measured. Differences in ultrasonic vocalization between HOA and LOA rats were detected during the conditioning day, where vocalization was more likely in LOA rats. Furthermore, LOA rats emitted calls with higher frequency components than HOA rats. On the subsequent day of testing, the number of animals vocalizing, and the rate of vocalization was decreased, and call differences between groups were no longer detectable. In freezing, differences between HOA and LOA rats were observed on the conditioning day, where LOA rats showed more freezing behavior during the tone/shock intervals. Also, on the test day, they showed more freezing behavior during the tone intervals compared to HOA rats. These results indicate that acute and conditioned responses of rats in a conventional fear conditioning paradigm can depend on individual dispositions of anxiety-related behavior as measured with the elevated plus-maze. In a 2nd experiment, we asked whether exposure to an elevated plus-maze would lead to ultrasonic vocalization, which we tested in rats which had been handled or non-handled prior to testing. Most importantly, we found that none of the animals displayed any vocalization in the plus-maze, neither during a 1st nor a repeated test 1 day later. These data are discussed with respect to the presumed role of ultrasonic vocalization in aversively motivated situations, and the mechanisms, which may account for the behavioral differences between HOA and LOA rats in such tests. PMID- 16213034 TI - Normal attention orienting but abnormal stimulus alerting and conflict effect in combined subtype of ADHD. AB - Recent pharmacological studies in animals and neuroimaging studies in normal humans suggest that the spatial and nonspatial cues in tasks measuring reflexive attention may be modulated by different neurotransmitter systems. The efficiency with which attention is oriented to explicit spatial cues may be altered by manipulating levels of brain acetylcholine, whereas reactions to nonspatial cues may be influenced by altering brain noradrenaline levels but not acetylcholine levels. In humans, however, previous attention studies have implicated dopamine when either explicit or implicit cueing is used. Some of the differences between animal and human work may be due to inadequate testing of nonspatial cues. To remedy this, we tested adult humans with ADHD that were primarily inattentive (ADHD/I) or combined inattentive/hyperactive (ADHD/C) and controls with the Attention Network Task that assesses both reflexive and voluntary attention and explicitly tests nonspatial cueing. Our results showed that spatial orienting in both subtypes was no different than controls. However, ADHD/C but not ADHD/I subjects had significantly slowed response times to nonspatial cues and cues with spatial conflict. Stimulant medication in a subset of ADHD/C subjects reduced these deficits to control levels. Based on these results, we conclude that ADHD/C subjects orient the focus of their attention normally but are impaired in their reactions both to abrupt visual cues and those that contain conflicting spatial cues. PMID- 16213035 TI - Aggression and vasotocin are associated with dominant-subordinate relationships in zebrafish. AB - Agonistic interactions are present throughout the animal kingdom as well as in humans. In this report, we present a model system to study neurological correlates of dominant-subordinate relationships. Zebrafish, Danio rerio, has been used as a model system for developmental biology for decades. We propose here that it is also an excellent model for studying social behavior. Adult male zebrafish were separated for 5 days and then pairs were formed and allowed to interact for 5 days. Under these conditions, aggression is prevalent and dominant subordinate relationships are quickly established. Dominant behavior is characterized by a repeated pattern of chasing and biting, whereas subordinates engage in retreats. By day 5, the dominant-subordinate relationship was firmly established and there were differences in behavior over time. Chases, bites and retreats were all less frequent on day 5 of the social interaction than on day 1. Arginine vasotocin is the teleostean homologue of arginine vasopressin, a neuropeptide whose expression has been linked to aggression and social position in mammals. Immunohistochemistry indicated differences in vasotocin staining between dominant and subordinate individuals. Dominant individuals express vasotocin in one to three pairs of large cells in the magnocellular preoptic area whereas subordinate individuals express vasotocin in 7-11 pairs of small cells in the parvocellular preoptic area. These results suggest that the vasotocinergic system may play a role in shaping dominant-subordinate relationships and agonistic behavior in this model organism. PMID- 16213036 TI - Metacaspases of Trypanosoma cruzi: possible candidates for programmed cell death mediators. AB - The genome of Trypanosoma cruzi, the Protozoan parasite causing the American Trypanosomiasis, Chagas disease, contains two genes, TcMCA3 and TcMCA5, with homology to those encoding metacaspases, distantly related to the caspases involved in programmed cell death (PCD) in higher eukaryotes. TcMCA3 is present in the CL Brener clone at 16 copies per haploid genome, arrayed in two tandems located in chromosomes of 0.54 and 0.98 Mbp. TcMCA5, on the other hand, is present as a single copy gene. The proteins encoded were expressed in Escherichia coli BL21 [DE3] cells, and used to generate antibodies, which allowed demonstrating that TcMCA3 is expressed in the four major developmental stages of the parasite, whereas TcMCA5 is expressed only in the epimastigote form. Moreover, recombinant TcMCA3, but not TcMCA5, was recognized by most sera from chronic Chagasic patients, showing that the protein is expressed during natural infections. All attempts to show processing and enzyme activity in the recombinant proteins have been unsuccessful so far; however, indirect evidence suggests that the metacaspases might be involved in PCD of the parasite. (1) Immunofluorescence experiments showed that both proteins change their subcellular localization during fresh human serum (FHS)-induced PCD migrating into the nucleus. (2) Epimastigotes over-expressing TcMCA5 were more sensitive to FHS induced PCD than the controls. (3) PCD was parallelled by an increase in peptidase activity against Z-YVAD-AFC, a typical caspase substrate, and the apoptotic nuclei cells were labeled in vivo with the pan-caspase fluorescent inhibitor SR-VAD-FMK. Further experiments will be required to complete the characterization of these proteins and elucidate their role in the parasite. PMID- 16213037 TI - Antral content, secretion and peripheral metabolism of N-terminal progastrin fragments. AB - OBJECTIVES: In addition to the acid-stimulatory gastrins, progastrin also release N-terminal fragments. In order to examine the cellular content, secretion and peripheral metabolism of these fragments, we developed an immunoassay specific for the N-terminal sequence of human progastrin. RESULTS: The concentration of N terminal progastrin fragments in human antral tissue was 6.7 nmol/g tissue (n=5), which was only half of that of acid-stimulatory gastrins (12 nmol/g tissue). Gel chromatography of antral extracts showed that the progastrin fragment 1-35 and 1 19 constitute the major part of the N-terminal progastrin fragments. The basal concentration of N-terminal fragments in normal human plasma was almost 30-fold higher than that of the amidated, acid-stimulatory gastrins (286 pmol/l versus 9.8 pmol/l, n=26, P<0.001). In contrast, the concentration of N-terminal fragments in hypergastrinemic plasma was only 2.7-fold higher than the concentration of amidated gastrins (540 pmol vs. 198 pmol/l, P=0.02). During meal stimulation, the plasma concentrations of N-terminal progastrin fragments and amidated gastrins increased in a correlated manner (r=0.97, P=0.005). The half life for progastrin 1-35 in circulation was 30 min, and a pig model revealed the kidneys and the vasculature to the head as the primary sites of degradation. CONCLUSION: The cellular and circulatory concentration profiles of N-terminal progastrin fragments differ markedly from those of the acid-stimulatory gastrins. The high basal plasma concentrations of N-terminal progastrin fragments cannot be explained by differences in elimination. PMID- 16213038 TI - Internalization of cloned pancreatic polypeptide receptors is accelerated by all types of Y4 agonists. AB - Internalization of cloned rat or human Y4 receptors expressed in Chinese hamster ovary (CHO) cells increased with concentration of all types of Y4 agonists, including human and rat pancreatic polypeptides, the Y1 receptor group co agonists possessing C-terminal TRPRY.NH2 pentapeptide, and a C-terminally amidated dimeric nonapeptide related to neuropeptide Y, GR231118. These peptides also inhibited forskolin-stimulated adenylyl cyclase activity in Y4 receptor expressing cells, and stimulated the binding of 35S-labeled GTP-gamma-S to pertussis toxin-sensitive G-proteins in particulates from these cells. Peptide VD 11 (differing from GR231118 only by C-terminal oxymethylation) acted as a competitive antagonist in all of the above processes. Agonist-induced stimulation of the Y4 receptor internalization persisted in the presence of allosteric inhibitors of hPP binding, N5-substituted amilorides, which also were relatively little active in G-protein stimulation and cyclase inhibition by Y4 agonists. Acceleration of Y4 receptor internalization by agonists apparently is related to relaxation of allosteric constraints to ligand attachment and sequestration of the receptor-ligand complex. PMID- 16213039 TI - Evaluation of in-hospital management of fracture risk in older patients: a chart review study of tertiary prevention. AB - Hip fractures are associated with considerable morbidity and mortality in the elderly. Both fall prevention strategies and bone integrity/osteoporosis assessment should be addressed in this population. This study's goal was to evaluate the management of potential re-fracture risk after a hip fracture in an acute care setting. This was a retrospective chart review of patients who were admitted with a hip fracture over the course of one year to the Ottawa Hospital, Civic Campus, Ottawa, Canada. The charts of 147 patients with hip fractures met the inclusion criteria. Use of sedatives on admission was significant (24.5%). Fifty (34%) had some form of osteoporosis management ordered during their hospital stay. The medication recommendations consisted of only 14% being prescribed Vitamin D and 15.6% being prescribed calcium supplementation. Merely 7 (4.8%) patients of the total sample were prescribed bisphosphonates at time of discharge. This study documents a significant care gap in re-fracture management at the time of acute hospitalization after an acute hip fracture. Interventions are required to increase the awareness that this problem is not being addressed at the time of hospitalization and that on discharge, patients will need follow up by the treating community physician. PMID- 16213041 TI - Control of BVDV-infection on common grassland--the key for successful BVDV eradication in Lower Austria. AB - A bovine viral diarrhea/mucosal disease (BVD/MD) control and eradication program was introduced in Lower Austria in 1996, according to the Swedish model. At present 9800 out of 17,000 herds are part of this program. An important risk factor for BVDV-transmission under local conditions is communal grazing. Approximately 3-4% of livestock share common pastures, in which susceptible pregnant cattle may be mixed with unrecognised persistently infected (PI) animals. Rules and regulations were defined to allow only herds free from BVDV infection on to common grassland. At the moment, 5067 herds are certified free from BVDV. The percentage of BVDV-free herds in regions with intensive pasture utilisation is higher (57.3%) than in the other regions (43.0%) of Lower Austria. With a reliable system for identification of PI-animals and a high certainty of prevention of PI-animals on common grassland, the main transmission of BVDV infection can be stopped, even if the animals are derived from infected herds and transiently infected animals cannot be excluded. PMID- 16213040 TI - Predicting readmissions and cardiovascular events in heart failure patients. AB - AIMS: To analyse measures of clinical data, functional capacity, left ventricular function and neurohormonal activation for the ability to predict mortality and morbidity in patients after a hospitalisation for heart failure. METHODS: In a prospective study, patients 60 years or above with systolic heart failure NYHA II IV were followed for at least 18 months. At study start, a physical examination, echocardiography, blood samples and measurements of quality of life (QoL) by Nottingham Health Profile were obtained. Data on mortality and readmission rates were collected. RESULTS: 208 patients, 58% men, with a mean age of 76 years, and an ejection fraction of 0.34 were included and followed for a mean of 1,122 days. In all, 74 (36%) patients died and 171 (82%) were readmitted. By univariate analysis, readmissions were predicted by poor QoL (169 +/- 118 vs. 83 +/- 100, p < 0.001), age, creatinine, haemoglobin (p < 0.01 all) and diabetes (p < 0.1). By multivariate analyses, QoL at study start was the only independent predictor of readmissions (chi(2) = 25.2, p < 0. 001). Mortality was univariately associated with QoL (183 +/- 117 vs. 142 +/- 115, p < 0.05) and in multivariate analyses to traditional variables: age, male gender, systolic function, BNP and serum creatinine (chi(2) = 48.9, p < 0.001). CONCLUSIONS: Measurements representing different aspects of the heart failure syndrome can easily be obtained to stratify long-term risks of mortality and morbidity in hospitalised heart failure patients. Poor QoL was a univariate predictor for mortality and a strong multivariate predictor for the important outcome of readmission, pointing to the need for a simple assessment of QoL. PMID- 16213042 TI - Pathophysiology of mental illness: a view from the fourth ventricle. AB - The concept of mental disorders as diseases of the brain is as old as the ancient Greek philosopher-physicians. However, for thousands of years, the majority of doctors, as well as laypersons, held strongly to the belief that epilepsy and "madness" (i.e., schizophrenia)--the major disorders discussed in this paper- were caused by demonic "possession." As always, the theory of causation led to apposite therapies: Cast out the devil by whatever means necessary. Later, more enlightened views of etiology led to less punitive "cures," which, to modern sensibilities, still seem barbaric. The 20th century saw the introduction of medications that provide symptomatic relief, if not cures, for seizure disorders and schizophrenia. In this paper, we consider the etiology of absence (petit mal) epilepsy and schizophrenia based on the pathophysiology underlying the shared symptom of impaired sustained attention. We emphasize the role of abnormal functioning of brainstem structures in the region of the fourth ventricle, whether caused by genetic or environmental factors or a combination thereof. Our theorizing relies on the findings of Lindsley, Magoun, and Moruzzi, who delineated the role of the brainstem reticular activating system in sleep, wakefulness, and consciousness. It also relies on the work of Penfield, Jasper, and Gloor, who sought to illuminate the role of brainstem-thalamus-cortical dysfunction in idiopathic generalized epilepsies. We consider evidence from recent studies that emphasize the phasic attentional functions supported by brainstem structures in the region of the fourth ventricle, and possible genetic links among disorders in which impaired attention is a prominent symptom. PMID- 16213044 TI - Oscillatory characteristics of face-evoked neuromagnetic responses. AB - To study the oscillatory activities during face processing, we recorded magnetoencephalographic responses in 8 healthy subjects to upright and inverted human faces, and obtained the time-frequency representation by using wavelet transform. Delta to beta activities were clearly increased at 140-210 ms after stimulus onset in the bilateral occipitotemporal (OT) areas (t(7)>5.5; p<0.001), with larger power for theta, alpha and beta over the right side. Notably, more increase alpha activity for inverted than upright face condition was observed in the right OT area. Our results suggest that 4-25 Hz oscillations are involved in face information processing, and the more activation over the right OT implies the right hemisphere advantage for face perception. Moreover, the alpha activity may reflect the differential cortical demands for processing inverted and upright face images. PMID- 16213045 TI - The transition from synchronization to continuation tapping. AB - The present study examined changes in timing at the transition from synchronization to continuation tapping and the role played by knowledge of the transition. Three experiments employed a pseudo-synchronization paradigm: At the transition, the pacing tones were replaced by identical feedback tones. In Experiment 1, participants were not informed when the transition would occur. Immediately following the transition, an acceleration of tapping was observed. In Experiment 2, participants were informed about the exact position of the transition, which was in addition marked by a pitch change. Nonetheless, the same acceleration occurred. Experiment 3 dissociated the actual from the expected position of the transition, without changing the results. In addition, the delay of the feedback tones was manipulated and was found to affect the rate of tapping in the continuation phase. A single delayed tone at the transition had no lasting effect, however. The results are interpreted in light of the synchronization continuation model of Vorberg and Wing [Vorberg, D., & Wing, A. (1996). Modelling variability and dependence in timing. In H. Heuer, & S. W. Keele (Eds.), Handbook of perception and action, Vol. 2 (pp. 181-262). San Diego: Academic Press], with the added assumption that the synchronization tapping strategy is maintained in the continuation phase. PMID- 16213043 TI - The hemodynamics of oddball processing during single-tone and two-tone target detection tasks. AB - Event-related potential (ERP) studies have shown that the neural systems engaged during performance of oddball tasks are sensitive to contextual manipulations, such as the number of stimulus classes. Some ERP components (i.e., N1) are modulated by the number of stimulus types, while others (i.e., P3) are not greatly affected. However, little is known about how these contextual manipulations affect the hemodynamics underlying oddball processing. The purpose of this study was to examine the hemodynamic correlates of target stimulus processing in single-tone (targets alone) and two-tone (targets and standard tones) auditory oddball tasks. The primary hypothesis was that processing of salient stimuli in both contextual conditions would engage the same spatially distributed cortical and subcortical networks observed in previous oddball fMRI studies. Results were consistent with this hypothesis and suggest that the brain engages many potentially useful brain regions during salient stimulus processing despite the low probability that said regions are necessary for task performance, which likely reflects a form of "adaptive reflexive processing". Results were also consistent with ERP data that shows that the N1 is larger for single-tone tasks by showing greater amplitude of hemodynamic response for single-tone targets, relative to two-tone targets, in bilateral temporal cortex and bilateral inferior lateral frontal cortex. The results are discussed as they relate to the understanding of neurocognitive function pertaining to contextual manipulations in general, and orienting processes in particular. PMID- 16213046 TI - Influence of swing leg movement on running stability. AB - The aim of this study was to investigate the role of the swing leg movement on running stability. A simple model was used describing a forward hopping motion. The model consisted of two sub-models, namely a spring-mass system for the stance phase and a functional control model for the swing phase (represented by a passive or actively driven pendulum). To verify the main simulation results, an experimental study on treadmill running was performed. The results of the model indicated that for certain running speeds and pendulum lengths, the behavior of the mechanical system was stable. The following characteristic dependencies between the model parameters were observed. (1) Pendulum length and hip muscle activity determined running height and therefore swing duration. (2) Horizontal velocity was inversely related to leg angle of attack. Increased speed corresponded to flatter leg angles at touch-down, which is in agreement with experimental studies and previous predictions of spring-mass running. It was shown that a biologically motivated control approach with oscillating leg movements is well capable of generating stable hopping movements. Due to its simplicity, however, the monopedal model failed to explain more detailed mechanisms like the swing-leg to stance-leg interaction or the functional role of the leg segmentation. This simple model is therefore considered as a functional mechanical template for legged locomotion, which could help to build more elaborate models in the future. PMID- 16213047 TI - Cloning and functional expression of the bovine GABA(C) rho2 subunit. Molecular evidence of a widespread distribution in the CNS. AB - GABA(C) receptors were first described as a non-desensitizing, bicuculline- and baclofen-insensitive component in Xenopus oocytes expressing bovine retina mRNA. However, the expression, tissue distribution and functional properties of GABA(C) receptors from other areas of the CNS still remain controversial. In previous experiments, the injection of rat cerebellum mRNA into Xenopus oocytes induced the expression of receptors that generated currents with both GABA(A) and GABA(C) characteristics; the latter component apparently being given by the rho2 subunit, suggesting the expression of GABA(C) receptors in the CNS and the formation of homooligomeric receptors. In this study, using RT-PCR, we found that the rho1 and rho2 subunits are widely expressed in the CNS including areas where they have not been previously described such as the bulb, pons and the caudate nucleus. To determine if the GABA(C) component of the GABA-currents elicited by oocytes expressing cerebellum mRNA was caused by activation of homomeric GABA rho2 receptors, we cloned the corresponding cDNA and expressed it in Xenopus oocytes. It was found that oocytes injected with rho2 cDNA, efficiently formed GABA-gated homooligomeric receptors. The GABA-dose-current response gave an EC50=1.19muM and the currents were resistant to bicuculline and reversibly antagonized by the specific GABA(C) receptor antagonist TPMPA. Altogether, our results indicate a widespread distribution of both rho1 and rho2 subunits in the bovine CNS and show further that the rho2 subunit cDNA isolated from cerebellum, forms fully functional receptors when expressed in Xenopus oocytes. PMID- 16213048 TI - Cortical mechanisms of unilateral voluntary motor inhibition in humans. AB - While motor control is very often a goal-oriented event, little is known about the mechanisms underlying the termination of motor performance. To investigate what type of cortical activation underlies the muscle relaxation required to terminate the act, we performed single- and double-pulse transcranial magnetic stimulation (TMS) studies during voluntary muscle relaxation in nine normal volunteers. Subjects maintained a weak isometric contraction of the right first dorsal interosseous muscle (FDI), and either increased the level of contraction (Contraction), terminated the contraction (Relaxation), or maintained it (No-go) depending on a visual cue. Motor evoked potentials (MEP) and the silent period (SP) were recorded from the FDI during motor activity. To measure intra-cortical inhibition (ICI), we also performed double-pulse TMS, applying subthreshold conditioning stimuli at interstimulus intervals of 2 ms. When single-pulse TMS was given just prior to muscle relaxation (-21 to -70 ms), the MEP was reduced while the SP was unchanged. Intra-cortical inhibition was smaller just prior to the muscle relaxation. Unilateral voluntary muscle relaxation may not be associated with activation of the intracortical inhibitory system, but rather with the possible excitation of the corticospinal system, which can inhibit motoneurons disynaptically. These findings suggest that multiple inhibitory mechanisms act in diverse ways to achieve motor inhibition. PMID- 16213049 TI - The effects of yeasts involved in the fermentation of Coffea arabica in East Africa on growth and ochratoxin A (OTA) production by Aspergillus ochraceus. AB - The effects of Pichia anomala, Pichia kluyveri and Hanseniaspora uvarum predominant during coffee processing on growth of Aspergillus ochraceus and production of ochratoxin A (OTA) on malt extract agar (MEA) and on coffee agar (CA) were studied. The three yeasts were able to inhibit growth of A. ochraceus when co-cultured in MEA and CA. Growth inhibition was significantly higher on MEA than on CA. Furthermore, P. anomala and P. kluyveri were found to have a stronger effect on growth of A. ochraceus than H. uvarum. The three yeasts were able to prevent spore germination of A. ochraceus in yeast glucose peptone (MYGP) broth. In yeast-free supernatant of MYGP broth after an incubation period of 72 h, spores of A. ochraceus were able to germinate with very short germ tubes, but further development of the germ tubes was inhibited. The three yeasts decreased the pH of MYGP broth from 5.6 to a range of 4.4-4.7, which was found to have no effect on spore germination of A. ochraceus. P. anomala, P. kluyveri and H. uvarum were able to prevent production of OTA by A. ochraceus when co-cultured on MEA. On CA medium, P. anomala and P. kluyveri prevented A. ochraceus from producing OTA. H. uvarum did not affect production of OTA by A. ochraceus on CA medium. PMID- 16213050 TI - Effects of yeasts and bacteria on the levels of folates in rye sourdoughs. AB - Fermentation of rye dough is often accompanied with an increase in folate content. In this study, three sourdough yeasts, Candida milleri CBS 8195, Saccharomyces cerevisiae TS 146, and Torulaspora delbrueckii TS 207; a control, baker's yeast S. cerevisiae ALKO 743; and four Lactobacillus spp., L. acidophilus TSB 262, L. brevis TSB 307, L. plantarum TSB 304, and L. sanfranciscensis TSB 299 originally isolated from rye sourdough were examined for their abilities to produce or consume folates. The microorganisms were grown in yeast extract peptone-d-glucose medium as well as in small-scale fermentations that modelled the sourdough fermentation step used in rye baking. Total folate contents were determined using Lactobacillus rhamnosus (ATCC 7469) as the growth indicator organism. The microorganisms studied did not excrete folates into the media in significant amounts. Yeasts increased the folate contents of sterilised rye flour water mixtures from 6.5 microg/100 g to between 15 and 23 microg/100 g after 19-h fermentation, whereas lactic acid bacteria decreased it to between 2.9 and 4.2 microg/100 g. Strains of Lactobacillus bulgaricus, L. casei, L. curvatus, L. fermentum, L. helveticus, Pediococcus spp., and Streptococcus thermophilus that were also tested gave folate contents after fermentation that varied between 2 and 10.4 microg/100 g. Although the four Lactobacillus spp. from sourdough consumed folates their effect on folate contents in co-cultivations was minimal. It was concluded that the increase of folate content during fermentation was mainly due to folate synthesis by yeasts. Fermentation of non-sterilised flour water mixtures as such resulted in three-fold increases in the folate contents. Two folate producing bacteria were isolated from the non-sterilised flour and identified as Enterobacter cowanii and Pantoea agglomerans. PMID- 16213051 TI - Effect of trisodium phosphate adaptation on changes in membrane lipid composition, verotoxin secretion, and acid resistance of Escherichia coli O157:H7 in simulated gastric fluid. AB - Escherichia coli O157:H7 (HEC), E. coli O157:H7 rpoS mutant (HEC-RM), and nonpathogenic E. coli (NPEC) were step-wise adapted to trisodium phosphate (TSP) by incubation in broths of increasing concentration, from 0% to 0.6%, at 37 degrees C for 24 h. After incubation at each concentration, each population was examined for acid resistance (D value) in simulated gastric fluid of pH 1.5, cell envelope membrane lipid composition, and intracellular and extracellular verotoxin concentrations. The ratio of cis-vaccenic acid (18:1omega7c) to palmitic acid (16:0) increased, indicating increased membrane fluidity with increasing TSP concentration up to 0.4%, but decreased at 0.6%. HEC and HEC-RM adapted at 0.4% TSP had the highest verotoxin concentrations of 1805 and 1879 ng/ml, respectively. In addition, with HEC the ratio of extracellular to intracellular verotoxin concentration decreased at higher TSP concentrations. In contrast, the ratio for HEC-RM increased at 0.4% TSP. HEC adapted to 0.4% TSP had the greatest survival in gastric fluid (58 min D value) among all treatments. For HEC, the increase in membrane fluidity was associated with increased acid resistance and extracellular verotoxin concentration for cells adapted to 0.4% TSP. In contrast, the increase in membrane fluidity was associated with decreased acid resistance of TSP adapted HEC-RM although the extracellular verotoxin concentration increased. Therefore, the deletion of the rpoS gene appeared to affect the changes in verotoxin concentration and acid resistance of TSP adapted E. coli O157:H7. PMID- 16213052 TI - Characterisation of the microflora of attieke, a fermented cassava product, during traditional small-scale preparation. AB - Attieke is a fermented cassava product consumed mainly in Cote d'Ivoire. The aim of this study was to characterise the attieke fermentation by examining products from 15 small-scale production sites at various stages of its preparation. For the preparation of attieke, fresh cassava is grated to a pulp and inoculated with 10% of a spontaneous traditional inoculum. The inocula contained aerobic mesophiles at mean numbers of 8.2 x 10(7) cfu/g and lactic and acetic acids at mean concentrations of 0.2% and 0.1%, respectively. The mean pH was 5.0. Lactic acid bacteria were the dominant microorganisms in cassava pulp throughout fermentation with the mean numbers being 1.2 x 10(9) cfu/g after 15 h. The identification to the species level of microorganisms from one representative attieke production of good quality showed that, at the start of fermentation, Leuconostoc mesenteroides subsp. mesenteroides was present in the highest numbers, accounting for 20% of all lactic acid bacteria. As the fermentation proceeded, this species was replaced by homofermentative lactic acid bacteria, Lactobacillus salivarius and Lactobacillus delbrueckii subsp. delbrueckii, present at 20% and 16%, respectively, and obligate heterofermentatives, Lactobacillus fermentum and Lactobacillus confusus at 12% and 10%, respectively, of total lactic acid bacteria in the flora at the end of fermentation. High numbers of acid-sensitive microorganisms, Bacillus circulans, Bacillus lentus, Enterobacter sakazakii, Enterobacter cloacae and Klebsiella pneumoniae subsp. pneumoniae, were transferred to the pulp in the inocula, but acidification to a mean pH of 4.4 with mean lactic and acetic acid concentrations of 0.59% and 0.2%, respectively, prevented their growth and reduced their numbers to less than 10(2) cfu/g at the end of fermentation. The mean numbers of Candida tropicalis, the main yeast present, remained relatively constant at about 10(5) cfu/g throughout attieke production. The mean numbers of aerobic mesophiles decreased to below 10(2) cfu/g as a result of the steaming process. The finished attieke had a mean pH of 4.4 and mean lactic and acetic acid concentrations of 0.6% and 0.1%, respectively. PMID- 16213053 TI - Functional meat starter cultures for improved sausage fermentation. AB - Starter cultures that initiate rapid acidification of the raw meat batter and that lead to a desirable sensory quality of the end-product are used for the production of fermented sausages. Recently, the use of new, functional starter cultures with an industrially or nutritionally important functionality is being explored. Functional starter cultures offer an additional functionality compared to classical starter cultures and represent a way of improving and optimising the sausage fermentation process and achieving tastier, safer, and healthier products. Examples include microorganisms that generate aroma compounds, health promoting molecules, bacteriocins or other antimicrobials, contribute to cured meat colour, possess probiotic qualities, or lack negative properties such as the production of biogenic amines and toxic compounds. The vast quantity of artisan fermented sausages from different origins represents a treasure chest of biodiversity that can be exploited to create such functional starter cultures. PMID- 16213054 TI - Resuscitation of the viable but non-culturable state of Salmonella enterica serovar Oranienburg by recombinant resuscitation-promoting factor derived from Salmonella Typhimurium strain LT2. AB - A gene encoding the resuscitation-promoting factor (Rpf) from Salmonella Typhimurium LT2 was cloned and characterized. The amino acid sequence encoded by S. Typhimurium LT2 rpf gene shares 24.2% homology with Micrococcus luteus Rpf, which is secreted by growing cells, and required to resuscitate from viable but non-culturable (VNC) state. The S. Typhimurium LT2 rpf gene is 696 bp long, and shared a conserved segment with Salmonella enterica serovar Oranienburg (99.4%). Recombinant Rpf (rRpf) proteins of S. Typhimurium LT2 after expression in E. coli BL21 harboring the pET15-b plasmid was approximately 25 kDa. Since S. Oranienburg cells are relatively quick to enter the VNC state just after incubating in the presence of 7% NaCl at 37 degrees C for 3 days, we evaluated the biological effect of rRpf by using S. Oranienburg VNC cells. The rRpf not only promoted proliferation but also induced resuscitation of VNC cells to the culturable state in a dose-dependent manner. Therefore, rRpf may be useful for detection of bacterial contaminants present in the VNC form in food samples and the environment. PMID- 16213055 TI - Structural and antigenic analysis of the yellow head virus nucleocapsid protein p20. AB - Yellow head virus (YHV) is an invertebrate nidovirus that is highly pathogenic for marine shrimp. Nucleotide sequence analysis indicated that the YHV ORF2 gene encodes a basic protein (pI = 9.9) of 146 amino acids with a predicted molecular weight of 16,325.5 Da. The deduced amino acid sequence indicated a predominance of basic (15.1%), acidic (9.6%) and hydrophilic polar (34.3%) residues and a high proportion proline and glycine residues (16.4%). The ORF2 gene was cloned and expressed in Escherichia coli as a M(r) = 21 kDa His(6)-protein that reacted with YHV nucleoprotein (p20) monoclonal antibody. Segments representing the four linear quadrants of the nucleoprotein were also expressed in E. coli as GST fusion proteins. Immunoblot analysis using YHV polyclonal rabbit antiserum indicated the presence of linear epitopes in all except the V(37)-Q(74) quadrant. Immunoblot analysis of the GST-fusion proteins and C-terminally truncated segments of the nucleoprotein allowed mapping of YHV monoclonal antibodies Y19, Y20 and YII4 to linear epitopes in the acidic domain between amino acids I(116) and E(137). The full-length nucleoprotein was expressed at high level in E. coli and was easily purified in quantity from the soluble cell fraction by Ni(+)-NTA affinity chromatography. PMID- 16213056 TI - Paclitaxel-loaded PLGA nanoparticles: potentiation of anticancer activity by surface conjugation with wheat germ agglutinin. AB - PURPOSE: To potentiate the anticancer activity of paclitaxel-loaded PLGA nanoparticles through surface conjugation with wheat germ agglutinin (WGA). METHODS: PLGA nanoparticles loaded with paclitaxel and isopropyl myristate (IPM) as release modifier were prepared by a solvent evaporation method. WGA was conjugated to the nanoparticle surface to give novel WIT-NP of 330+/-3 nm. In vitro cytotoxicity of WIT-NP against malignant (A549 and H1299) and normal (CCL 186) pulmonary cell lines was evaluated alongside control formulations. IC50 doses were determined by the MTT assay, while cellular apoptosis was detected by cell nuclei staining and DeadEndtrade mark Fluorometric TUNEL assay. Cell cycle arrest was confirmed by flow cytometry. Cellular uptake of 3[H]-paclitaxel from the test and control formulations was also quantified. In vivo anticancer efficacy was evaluated in the SCID mice model engrafted with the A549 tumor nodule. RESULTS: WIT-NP had superior anti-proliferation activity against the A549 and H1299 cell lines compared with conventional paclitaxel formulations as measured by IC50 doses. This was attributed to a more efficient intracellular accumulation of paclitaxel via WGA-receptor-mediated endocytosis and IPM facilitated intracellular paclitaxel release. WIT-NP activity was associated with paclitaxel-induced apoptosis and cell arrest in the G2/M phase. A single intratumoral injection of WIT-NP at paclitaxel dose of 10 mg/kg inhibited the growth of A549 tumor nodules without inducing significant weight loss in the SCID mice over a period of 25 days. Tumor doubling time was greater than 25 days, compared with 11 days for nodules treated with conventional paclitaxel formulation. CONCLUSION: The formulation of WIT-NP, in which WGA is conjugated to the surface of paclitaxel and IPM-loaded PLGA nanoparticles, significantly potentiates the anticancer activity of paclitaxel. PMID- 16213058 TI - Genome size is negatively correlated with effective population size in ray-finned fish. AB - A recent theory suggesting that genome size and complexity can increase as a passive consequence of small effective population size has generated much controversy. In this article, we demonstrate that freshwater fish species, which have smaller effective population sizes than marine fish species, have larger genomes. We show that genome size is negatively correlated with genetic variability, independent of phylogeny, body size and generation time. Genome duplication is also observed predominantly in freshwater fish. These results suggest that the raw materials of complexity originate under conditions of reduced selection efficiency. PMID- 16213057 TI - Impact of the intragastric location of extended release tablets on food interactions. AB - Gastrointestinal motility and transport as well as concomitant food intake are factors that are known to influence pharmacokinetics derived after intake of extended release dosage forms. However, the mechanisms behind these influencing factors are mostly unknown. In this study the gastrointestinal transit and the in vivo drug release of magnetically labelled extended release tablets containing felodipine were monitored together with the drug absorption phase of pharmacokinetics under fasting and fed conditions in six healthy volunteers using Magnetic Marker Monitoring. It was found that the in vivo drug release profiles of the tablets compared well under fasting and fed conditions. However, the plasma concentration profiles were strongly influenced by concomitant food intake. This could be attributed to elongated residence of the tablets in proximal parts of the stomach, resulting in delayed drug absorption and the occurrence of late high plasma peak concentrations. The lag time until the first appearance of felodipine in plasma and the residence time of the tablets in the proximal stomach, were found to be directly correlated. The study shows that increased plasma peak drug concentrations after intake of extended release formulations together with food can be explained by poor mixing in the proximal part of the stomach and are not necessarily due to failure of the formulation to control drug release (dose dumping). PMID- 16213059 TI - Stereotactic hypofractionated radiotherapy for stage I non-small cell lung cancer -mature results for medically inoperable patients. AB - Medically inoperable patients with stage I NSCLC are mainly offered conventionally fractionated radiotherapy with a limited chance of local control and some toxicity. A technique for stereotactic precision therapy for extracranial tumors using a linear accelerator and a body frame for patient immobilization was applied in an attempt to improve the local control and decrease toxicity for consecutive patients with inoperable stage I NSCLC at Sahlgrenska University hospital since 1998. A hypofractionated schedule with three fractions of 15Gy to a total of 45 Gy during 1 week was used which represents a biological equivalent dose (BED) of 112.5 Gy. Planning target volume (PTV) was a 5mm margin around the tumor in the transversal plane and 10mm in the cranial-caudal direction and the dose was prescribed in the periphery of the PTV. Forty-five patients were treated between September 98 and March 03, 25 men and 20 women, median age 74 years (58-84) and median Karnofsky 80 (100-60). TNM: 18 T1N0, 27 T2N0. HISTOLOGY: 18 squamous cell carcinoma, 15 adenocarcinoma, 3 NSCLC and histology was missing in nine patients. The majority, 51%, did not experience any toxicity at all, four had esophagitis grade I, nine had skin reactions, four had transient chest pain and four had infections. Late toxicity was two rib fractures and three patients with atelectasias. After a median follow-up of 43 months had nine patients developed local recurrence or never achieved local control, two had regional recurrence and nine distant metastases. The 1-, 2-, 3- and 5-year overall survival was 80, 71, 55 and 30%, respectively, with a median survival of 39 months. No prognostic factor for survival could be identified among histology, tumor stage and size, gender and age. We think this hypofractionated stereotactic radiotherapy shows encouraging survival and a relatively low toxicity in this elderly population with substantial comorbidity. A multicenter randomized trial comparing this treatment with conventional fractionated radiotherapy is under way. PMID- 16213060 TI - Design of aquifer remediation systems: (1) describing hydraulic structure and NAPL architecture using tracers. AB - Aquifer heterogeneity (structure) and NAPL distribution (architecture) are described based on tracer data. An inverse modelling approach that estimates the hydraulic structure and NAPL architecture based on a Lagrangian stochastic model where the hydraulic structure is described by one or more populations of lognormally distributed travel times and the NAPL architecture is selected from eight possible assumed distributions. Optimization of the model parameters for each tested realization is based on the minimization of the sum of the square residuals between the log of measured tracer data and model predictions for the same temporal observation. For a given NAPL architecture the error is reduced with each added population. Model selection was based on a fitness which penalized models for increasing complexity. The technique is demonstrated under a range of hydrologic and contaminant settings using data from three small field scale tracer tests: the first implementation at an LNAPL site using a line-drive flow pattern, the second at a DNAPL site with an inverted five-spot flow pattern, and the third at the same DNAPL site using a vertical circulation flow pattern. The Lagrangian model was capable of accurately duplicating experimentally derived tracer breakthrough curves, with a correlation coefficient of 0.97 or better. Furthermore, the model estimate of the NAPL volume is similar to the estimates based on moment analysis of field data. PMID- 16213061 TI - Amygdala kindling differentially regulates the expression of the elements involved in TRH transmission. AB - Subthreshold electrical stimulation of the amygdala (kindling) activates neuronal pathways increasing the expression of several neuropeptides including thyrotropin releasing-hormone (TRH). Partial kindling enhances TRH expression and the activity or its inactivating ectoenzyme; once kindling is established (stage V), TRH and its mRNA levels are further increased but TRH-binding and pyroglutamyl aminopeptidase II (PPII) activity decreased in epileptogenic areas. To determine whether variations in TRH receptor binding or PPII activity are due to regulation of their synthesis, mRNA levels of TRH receptors (R1, R2) and PPII were semi quantified by RT-PCR in amygdala, frontal cortex and hippocampus of kindled rats sacrificed at stage II or V. Increased mRNA levels of PPII were found at stage II in amygdala and frontal cortex, and of pro-TRH and TRH-R2, in amygdala and hippocampus. At stage V, pro-TRH mRNA levels increased and those of PPII, decreased in the three regions; TRH-R2 mRNA levels diminished in amygdala and frontal cortex and of TRH-R1 only in amygdala. In situ hybridization analyses revealed, at stage II, enhanced TRH-R1 mRNA levels in dentate gyrus and amygdala while decreased in piriform cortex; those of TRH-R2 increased in amygdala, CA2, dentate gyrus, piriform cortex, thalamus and subiculum and of PPII, in CAs and piriform cortex. In contrast, at stage V decreased expression of TRH-R1 occurred in amygdala, CA2/3, dentate gyrus and piriform cortex; of TRH-R2 in CA2, thalamus and piriform cortex, and of PPII in CA2, and amygdala. The magnitude of changes differed between ipsi and contralateral side. These results support a trans synaptic modulation of all elements involved in TRH transmission in conditions that stimulate the activity of TRHergic neurons. They show that reported changes in PPII activity or TRH-binding caused by kindling relate to regulation of the expression of TRH receptors and degrading enzyme. PMID- 16213062 TI - Fitness, aging and neurocognitive function. AB - In this manuscript we provide a brief review of the recent literature that has examined the relationship among fitness training, cognition and brain. We began with a discussion of the non-human animal literature that has examined the relationship among these factors. Next we discuss recent epidemiological studies of the relationship between physical activity and fitness and cognition and age related disease such as Alzheimer's dementia. We then discuss the results of randomized clinical trials of fitness training on human cognition. Finally, we conclude with a review of the nascent literature that has begun to employ neuroimaging techniques to examine fitness training effects on human brain. In general, the results are promising and suggest that fitness may serve a neuroprotective function for aging humans. PMID- 16213063 TI - Investigating spiritual care perceptions and practice patterns in Hong Kong nurses: results of a cluster analysis. AB - AIM: Nurses' spiritual care perceptions and practices are explored by identifying profiles of nurses studying in a part-time baccalaureate course in a local Hong Kong university. Relationships between nurses' spiritual care perceptions and their practices are explored. RESEARCH METHOD: Hundred and ninety three nurses completed a structured questionnaire. OUTCOME MEASURES: Spiritual care perceptions and practices. RESULTS: Two-step cluster analysis yielded three clusters. Clusters A, B, and C consisted of 15.0% (n = 29), 44.6% (n = 86), and 40.4% (n = 78), respectively. Cluster A nurses were characterized by relatively negative spiritual care perceptions and practices. Cluster C nurses reported positive perceptions, but negative practices; they mainly chose 'uncertain' for most items on both scales. Cluster B was a large group of nurses holding both positive spiritual care perceptions and practices. Significant differences towards spiritual care were found among clusters. Nurses' perceptions were significant positively correlated with practices (r = 0.62). High positive correlations were found between the two scales (r = 0.83) for nurses in Cluster A, for nurses in Clusters B and C, low positive correlations (r = 0.37) were found. CONCLUSION: Three clusters of Hong Kong nurses were differentiated. They showed differences in the level of their spiritual care perceptions and practices. Despite their level of spiritual care perceptions, nurses seldom incorporated spiritual care practices into their daily nursing care, and the level of spiritual care awareness of some nurses was low. Findings may be used to improve support of nurses, to ensure sensitive spiritual care in their daily practices, and to enhance nursing curricula. PMID- 16213064 TI - Malnutrition determined by the patient-generated subjective global assessment is associated with poor outcomes in acute stroke patients. AB - BACKGROUND & AIMS: The extent of malnutrition in hospitalised stroke patients and its influence on outcomes including hospital complications, length of stay and discharge destination are important issues. The aim of this study was to determine the nutritional status of patients admitted to an acute stroke unit and the association between nutritional status and health outcomes. METHODS: Nutritional status was determined prospectively using the scored patient generated subjective global assessment (PG-SGA) in patients (n=73) admitted to an acute stroke unit within 48 h of admission to an Australian private hospital. Outcome data were collected by retrospective audit. RESULTS: On admission, 19.2% of patients were malnourished and this was associated with a significantly greater PG-SGA score (15 vs. 5) and lower body weight (59.8 kg vs. 75.8 kg) compared to well-nourished patients. In terms of health outcomes, malnourished patients had longer length of stay (13 vs. 8 days), increased complications (50% vs. 14%), increased frequency of dysphagia (71% vs. 32%) and enteral feeding (93% vs. 59%). No association was found between nutritional status and serum albumin level or discharge destination. CONCLUSIONS: Malnutrition on admission to hospital after acute stroke is associated with poor outcomes including increased length of stay and increased prevalence of dysphagia and complications. The scored PG-SGA is a nutrition assessment tool that allows quick identification of malnourished stroke patients. PMID- 16213065 TI - Antibody response to influenza vaccination in the elderly: a quantitative review. AB - We performed a quantitative review of 31 vaccine antibody response studies conducted from 1986 to 2002 and compared antibody responses to influenza vaccine in groups of elderly versus younger adults. We did a weighted analysis of the probability of vaccine response (measured as seroconversion and seroprotection) for each vaccine component (H1, H3 and B antigens). Using a multiple regression model, we adjusted for factors that might affect the vaccine response. The adjusted odds-ratio (OR) of responses in elderly versus young adults ranged from 0.24 to 0.59 in terms of seroconversion and seroprotection to all three antigens. The CDC estimates of 70-90% clinical vaccine efficacy in young adults and these estimates suggest a corresponding clinical efficacy in the elderly of 17-53% depending on circulating viruses. We conclude that the antibody response in the elderly is considerably lower than in younger adults. This highlights the need for more immunogenic vaccine formulations for the elderly. PMID- 16213066 TI - A randomized controlled trial of a replication defective (gH deletion) herpes simplex virus vaccine for the treatment of recurrent genital herpes among immunocompetent subjects. AB - BACKGROUND: A replication incompetent herpes virus lacking the glycoprotein H gene has been developed as a potential therapeutic vaccine for genital herpes. GOAL: To determine vaccine efficacy on reducing HSV reactivation and clinical disease among immunocompetent persons with recurrent genital HSV-2 infection. STUDY DESIGN: Randomized multicenter placebo-controlled trial. Healthy volunteers who had six or more recurrences of genital herpes per year were randomized to receive injections of vaccine at 0 and 8 or 0, 4, and 8 or 0, 2, 4, and 8 weeks or placebo and were followed for subsequent recurrences for 1 year. RESULTS: The median times to first recurrence of genital herpes (40 days versus 30 days versus 37 days versus 42 days, respectively), mean number of recurrences (3 versus 3 versus 2.4 versus 1.9, respectively), and time to lesion healing of the first recurrence (8 days versus 7.8 days versus 7.4 days versus 7.5 days, respectively), were similar for all treatment groups. Asymptomatic viral shedding was detected by PCR in 61/74 (82%) persons performing daily sample collection following completion of the vaccination series. No differences were noted in the proportion of days with shedding between treatment groups (11.9% versus 17.2% versus 13.1% versus 16.4%, respectively). CONCLUSION: This replication incompetent HSV-2 vaccine lacking the glycoprotein H gene was safe but had no clinical or virologic benefit in the amelioration of genital HSV-2 disease among immunocompetent men and women. PMID- 16213067 TI - Resuspension and redistribution of radionuclides during grassland and forest fires in the Chernobyl exclusion zone: part I. Fire experiments. AB - Controlled burning of experimental plots of forest or grassland in the Chernobyl exclusion zone has been carried out in order to estimate the parameters of radionuclide resuspension, transport and deposition during forest and grassland fires and to evaluate the working conditions of firemen. An increase of several orders of magnitude of the airborne radionuclide concentration was observed in the territory near the fire area. The resuspension factor for (137)Cs and (90)Sr was determined to range from 10(-6) to 10(-5) m(-1), and for the plutonium radionuclides from 10(-7) to 10(-6) m(-1) (related to the nuclides in the combustible biomass). These values are 2 orders of magnitude lower if they are calculated relatively to the total contamination density (including the nuclides in the soil). The radionuclide fallout along the plume axis is negligible in comparison to the existing contamination. However, the additional inhalation dose for firemen exposed in the affected area can reach the level of the additional external irradiation in the period of their mission. The plutonium nuclides constitute the dominating contribution to the inhalation dose. PMID- 16213068 TI - Vertical profiles and enrichment pattern of natural radionuclides in monazite areas of coastal Kerala. AB - Detailed studies on radionuclides concentration in different environmental matrices of high background areas were undertaken in the coastal areas of Karunagapalli, Chavara, Neendakara and Kollam to study the distribution and enrichment of the radionuclides in the region. The sand samples collected at different distances from sea waterline and at different depths, were analysed for primordial radionuclides by gamma spectrometry. The activity of primordial radionuclides was determined for the different size fractions of sand to study the enrichment pattern. The highest activity was found confined in 125-63 microm particle size fraction in sand. The minimum (232)Th activity was 9.4 Bq kg(-1), found in Kollam at a depth of 10-20 cm, 40 m away from waterline in 500-250 microm particle size fraction and maximum activity of 136,811.2 Bq kg(-1) was observed in Chavara in grains of size 125-63 microm at a depth of 0-10 cm for a sample collected 20 m away from waterline. The lowest (226)Ra activity observed was 29.6 Bq kg(-1) at Kollam beach for a sample 40 m away from waterline in grains of size 1000-500 microm and at a depth of 20-30 cm and the highest activity observed was 10,309 Bq kg(-1) in grains of size 125-63 microm for a sample collected at a distance 20 m away from waterline and at a depth of 0-10 cm. The activity of (40)K was below detectable level in most of the samples collected from the high background monazite area. The (232)Th, (226)Ra activities decrease with depth for the samples collected 20 m away from the waterline and increase with depth for the samples collected 40 m away from the waterline at Chavara and Kollam beaches. No definite correlation was found between variation of the concentrations of (232)Th and (226)Ra with depth at Karunagapalli and Neendakara beach sands. There exists a strong correlation between (226)Ra and (232)Th activities in the region. The results of these investigations are presented and discussed in this paper. PMID- 16213069 TI - Symptomatic venous thromboembolism after orthognathic operations: an audit. AB - We report the incidence of symptomatic venous thromboembolism confirmed by venography after 129 consecutive orthognathic operations between 1998 and 2002. Two patients developed deep vein thromboses (DVT) and there were no symptomatic pulmonary emboli. PMID- 16213070 TI - Kimura disease in an African patient. AB - We operated on a 23-year-old black Nigerian man with a 4-year history of a tumour on the left cheek associated with IgE hypergammaglobulinaemia and peripheral eosinophilia. The lesion recurred. PMID- 16213071 TI - Recovery of the histogram of hourly ozone distribution from weekly average concentrations. AB - A simple method is presented for estimating hourly distribution of air pollutants, based on data collected by passive sensors on a weekly or bi-weekly basis with no need for previous measurements at a site. In order for this method to be applied to locations where no hourly records are available, reference data from other sites are required to generate calibration histograms. The proposed procedure allows one to obtain the histogram of hourly ozone values during a given week with an error of about 30%, which is good considering the simplicity of this approach. This method can be a valuable tool for sites that lack previous hourly records of pollutant ambient concentrations, where it can be used to verify compliance with regulations or to estimate the AOT40 index with an acceptable degree of exactitude. PMID- 16213073 TI - Stable isotope dynamics of nitrogen sewage effluent uptake in a semi-arid wetland. AB - Our objectives were to determine (1) how much N is transferred into the food web via plants from a wetland receiving not only inputs of treated sewage effluent, but also containing contaminants such as polychlorinated biphenyls (PCBs), (2) how birds, as consumers, utilize exogenous N and uptake PCBs in relation to the food web of the wetlands, (3) the feasibility of using isotopic analysis in estimating trophic levels in a semi-arid system. Our results demonstrate that there is very high spatial variability in the N isotopic composition of primary producers. Birds had lower variability in delta15N, despite feeding at multiple trophic levels. In very high spatial variability in delta15N of primary producers, it is difficult to use N isotope techniques to define trophic levels relevant to the bioaccumulation of organic pollutants, but it is possible to track the flow of exogenous N through the food web. PMID- 16213072 TI - Effect of chromium on the fatty acid composition of two strains of Euglena gracilis. AB - The effect of hexavalent chromium on fatty acid composition was studied in two strains of Euglena gracilis; UTEX 753 (from the Culture Collection of Algae of Texas University, USA) and MAT (isolated from a highly polluted River). Both were grown in photoauxotrophic and photoheterotrophic conditions and exposed to two metal concentrations, one below and one above IC50. The high malondialdehyde (MDA) levels (3 to 7-fold) obtained with chromium concentration above IC50, suggested the existence of metal-induced lipid peroxidation. Total lipid content increased only with concentration below IC50, whereas it was inhibited by higher metal concentration. Photoheterotrophic control strains exhibited a significantly higher proportion of saturated and polyunsaturated fatty acids. Polyunsaturated acids were most affected by chromium, especially those related to chloroplast structures. Ultra-structure studies showed clear thylakoid disorganization in all treated cells. The results indicate that hexavalent chromium affects levels of fatty acids, especially those related to photosynthetic activity. PMID- 16213074 TI - Application of the threshold of toxicological concern approach to ingredients in personal and household care products. AB - In the absence of chemical-specific data, the threshold of toxicological concern (TTC) provides a method to determine a conservative estimate of a chronic oral exposure below which there is a very low probability of risk. The TTC approach was originally developed to support exposures to indirect food additives and was based on linear low-dose risk estimates to assure protection in the event that the chemical was later determined to be a carcinogen. Subsequently, TTC values based on noncancer endpoints were proposed for chemicals without structural alerts for genotoxicity. The original database supporting the TTC values for noncancer endpoints includes >600 structurally diverse chemicals. The objectives of this work were to evaluate the applicability of the TTC database to ingredients used in consumer products based on a comparison of the diversity of chemical structures with those in the original TTC database and to confirm that the range of NOELs for these ingredients is consistent with the range of NOELs in the original database. The results show good coverage of the product ingredient structures and confirm that the NOELs for the ingredient chemicals are similar in range to the original dataset, supporting the use of the TTC for ingredients in consumer products. PMID- 16213075 TI - The role of science in international trade law. AB - While the General Agreement on Tariffs and Trade addressed overt barriers to international trade, the current focus of international trade rules has shifted to less obvious, but in many cases no less restrictive, barriers to trade, such as protectionist measures adopted under the guise of health and safety standards. The new agreements established under the World Trade Organization ("WTO"), including the Agreement on the Application of Sanitary and Phytosanitary Measures ("SPS Agreement"), the Agreement on Technical Barriers to Trade ("TBT Agreement"), provide important tools that can be invoked by governments and used by stakeholders to address regulatory barriers that were once thought outside the purview of international trade rules. Non-science based regulations can be and have been successfully challenged under the SPS and TBT Agreements, which prohibit WTO Members from maintaining laws or regulations that adversely affect trade unless such measures are scientifically justified. Stakeholders should use to the fullest extent possible international trade rules to eliminate non-science based regulations that adversely affect trade in the goods that they produce. PMID- 16213076 TI - Science and norms in policies for sustainable development: assessing and managing risks of chemical substances and genetically modified organisms in the European Union. AB - Use of chemical substances and genetically modified organisms cause complex problems characterised by scientific uncertainty and controversies. Aiming at sustainable development, policies for assessment, and management of risks in the two areas are under development in the European Union. The article points out that both science and norms play a central role in risk assessment as well as risk management and suggests that the precautionary principle, the principle of public participation, and the polluter pays principle, all adopted in the European Union, offer a way to operationalise the concept of sustainable development. It is shown, however, that a number of steps ought to be taken to better implement the principles through different policy measures. In doing so, and by recognising the role of both science and norms, the decision-making on risks related to the use of chemicals or genetically modified organisms can be improved to better promote sustainable development. PMID- 16213077 TI - Parents' experiences of sharing neonatal information and decisions: consent, cost and risk. AB - This paper is about the care of babies with confirmed or potential neurological problems in neonatal intensive care units. Drawing on recent ethnographic research, the paper considers parents' experiences of sharing information and decisions with neonatal staff, and approaches that support or restrict parents' involvement. There are growing medico-legal pressures on practitioners to inform parents and involve them in their babies' care. Data are drawn from observations in four neonatal units in southern England, and interviews with the parents of 80 babies and with 40 senior staff. The paper compares standards set by recent guidance, with parents' views about their share in decision-making, their first meetings with their babies, 'minor' decision-making, the different neonatal units, being a helpless observer and missed opportunities. Parents' standards for informed decisions are summarised, with their reported views about two-way decision-making, and their practical need to know. Whereas doctors emphasise distancing aspects of the consent process, parents tend to value 'drawing together' aspects. PMID- 16213079 TI - Defining stress as a prelude to mapping its neurocircuitry: no help from allostasis. AB - The way in which researchers conceptualise and thus define stress shapes the way in which they approach the task of mapping the brain's stress control pathways. Unfortunately, much of the research currently being done on stress neurocircuitry is occurring within a poorly developed conceptual framework, a framework that limits the depth of the questions that our studies ask, and even our ability to fully appreciate and make use of the data that they yield. Consequently, any attempt to improve our conceptual framework merits close attention. In that regard it is notable that in recent years it has been argued that the concept of homeostasis should be supplemented by the concepts of allostasis (literally 'stability through change') and allostatic load (in effect, the cost of allostasis). One of the purported benefits of this change has been that it will clarify the concept of stress. A close review of the arguments leads us to conclude that the introduction of the concept of allostasis has largely occurred as a result of misunderstandings and misapprehensions concerning the concept of homeostasis. In terms of understanding how the organism operates, it is not clear that the concepts of 'allostasis' or 'allostatic load' offer us anything that was not already apparent, or at least readily derivable, from an accurate reading of the original concept of homeostasis. Not surprisingly then, these more recently proposed concepts also offer little help in clarifying our understanding of stress. Indeed, rather than clarifying the concept of stress, the primary effort appears to be directed at subsuming the concept of stress within the concept of allostasis, which has the inadvertent effect of collapsing the study of homeostatic responses and stress responses together. This seems to be out of step with the fact that there is now considerable evidence that the brain does indeed possess certain pathways that merit the title of 'stress neurocircuitry'. The attempt to subsume the concept of stress within the concept of allostasis is also counter-productive in that it distracts stress researchers from the important task of developing conceptual frameworks that allow us to tackle fundamental issues such as how the organism differentiates stressful from non-stressful challenges. PMID- 16213078 TI - Associations of poverty, substance use, and HIV transmission risk behaviors in three South African communities. AB - The majority of the world's HIV infections occur in communities ravished by poverty. Although HIV/AIDS and poverty are inextricably linked, there are few studies of how poverty-related stressors contribute to HIV risk behavior practices. In this study, surveys were conducted in three South African communities that varied by race and socio-economic conditions: people living in an impoverished African township (N = 499); an economically impoverished but well infrastructured racially integrating township (N = 995); and urban non impoverished neighborhoods (N = 678). Results showed that HIV/AIDS risks were closely related to experiences of poor education, unemployment, discrimination, violence, and crime. Although poverty-related stressors were associated with a history of alcohol and drug use, substance use did not moderate the association between poverty-related stressors and HIV risk behaviors. The findings suggest that HIV prevention strategies should not treat AIDS as a singled out social problem independent of other social ills. PMID- 16213080 TI - Application of support vector machine (SVM) for prediction toxic activity of different data sets. AB - As a new method, support vector machine (SVM) were applied for prediction of toxicity of different data sets compared with other two common methods, multiple linear regression (MLR) and RBFNN. Quantitative structure-activity relationships (QSAR) models based on calculated molecular descriptors have been clearly established. Among them, SVM model gave the highest q(2) and correlation coefficient R. It indicates that the SVM performed better generalization ability than the MLR and RBFNN methods, especially in the test set and the whole data set. This eventually leads to better generalization than neural networks, which implement the empirical risk minimization principle and may not converge to global solutions. We would expect SVM method as a powerful tool for the prediction of molecular properties. PMID- 16213082 TI - Ion hydration: Implications for cellular function, polyelectrolytes, and protein crystallization. AB - Only oppositely charged ions with matching absolute free energies of hydration spontaneously form inner sphere ion pairs in free solution [K.D.Collins, Ions from the Hofmeister series and osmolytes: effects on proteins in solution and in the crystallization process, Methods 34 (2004) 300-311.]. We approximate this with a Law of Matching Water Affinities which is used to examine the issues of (1) how ions are selected to be compatible with the high solubility requirements of cytosolic components; (2) how cytosolic components tend to interact weakly, so that association or dissociation can be driven by environmental signals; (3) how polyelectrolytes (nucleic acids) differ from isolated charges (in proteins); (4) how ions, osmolytes and polymers are used to crystallize proteins; and (5) how the "chelate effect" is used by macromolecules to bind ions at specific sites even when there is a mismatch in water affinity between the ion and the macromolecular ligands. PMID- 16213083 TI - Neurophysiological correlates of age-related changes in working memory capacity. AB - Cognitive abilities such as working memory (WM) capacity decrease with age. To determine the neurophysiological correlates of age-related reduction in working memory capacity, we studied 10 young subjects (<35 years of age; mean age=29) and twelve older subjects (>55 years of age; mean age=59) with whole brain blood oxygenation-level dependent (BOLD) fMRI on a 1.5 T GE MR scanner using a SPIRAL FLASH pulse sequence (TE=24 ms, TR=56 ms, FA=60 degrees , voxel dimensions=3.75 mm(3)). Subjects performed a modified version of the "n" back working memory task at different levels of increasing working memory load (1-Back, 2-Back and 3 Back). Older subjects performed as well as the younger subjects at 1-Back (p=0.4), but performed worse than the younger subjects at 2-Back (p<0.01) and 3 Back (p=0.06). Older subjects had significantly longer reaction time (RT) than younger subjects (p<0.04) at all levels of task difficulty. Image analysis using SPM 99 revealed a similar distribution of cortical activity between younger and older subjects at all task levels. However, an analysis of variance revealed a significant group x task interaction in the prefrontal cortex bilaterally; within working memory capacity, as in 1-Back when the older subjects performed as well as the younger subjects, they showed greater prefrontal cortical (BA 9) activity bilaterally. At higher working memory loads, however, when they performed worse then the younger subjects, the older subjects showed relatively reduced activity in these prefrontal regions. These data suggest that, within capacity, compensatory mechanisms such as additional prefrontal cortical activity are called upon to maintain proficiency in task performance. As cognitive demand increases, however, they are pushed past a threshold beyond which physiological compensation cannot be made and, a decline in performance occurs. PMID- 16213084 TI - Functional characteristics of H+ -dependent nicotinate transport in primary cultures of astrocytes from rat cerebral cortex. AB - In the present study, we report the characteristics of H(+)-coupled nicotinate transport in primary cultures of astrocytes from rat cerebral cortex. The [(3)H]nicotinate transport in rat astrocytes increased up to a pH 5.5. The nicotinic acid uptake at pH 6.0 was both energy-dependent and saturable with a Michaelis constant (K(t)) of 2.8+/-0.4 mM and the maximal uptake rate (V(max)) of 31+/-3.2 nmol/mg protein/10 min. This process was reduced by a protonophore, carbonylcyanide p-trifluoromethoxyphenylhydrazone, and a typical monocarboxylate transporter (MCT) inhibitor, alpha-cyano-4-hydroxycinnamic acid, suggesting that nicotinate uptake by rat astrocytes is mediated by H(+)-coupled monocarboxylate transport system. [(3)H]Nicotinate transport in rat astrocytes was significantly inhibited by various monocarboxylic acids such as l-lactic acid and pyruvic acid with a relatively low affinity (K(i)>10 mM). On the other hand, the uptake process of l-lactic acid was also saturable with a high-affinity component (K(t)=0.27 mM) and a low-affinity component (K(t)=35.9 mM). Reverse transcription PCR and Western blot analyses revealed that three MCT subtypes, MCT1/Slc16a1, MCT2/Slc16a7, and MCT4/Slc16a3, were expressed in these cells. Because l-lactate reduced to 67% of the nicotinate uptake even at 10mM, it is unlikely that nicotinate uptake in rat astrocytes is mediated by MCT1 and/or MCT2. These results provide biochemical evidence of a H(+)-coupled and saturable transport system, presumed to be a low-affinity monocarboxylate transporter MCT4 or other unknown H(+)-coupled monocarboxylate transport system, for nicotinate in rat cerebrocortical astrocytes. PMID- 16213085 TI - TRPV4 mediates pain-related behavior induced by mild hypertonic stimuli in the presence of inflammatory mediator. AB - The ligand-gated ion channel, TRPV4, functions as a transducer of hypotonic stimuli in primary afferent nociceptive neurons and contributes to inflammatory and neuropathic pain. Hypertonic saline also stimulates primary afferent nociceptors and the injection of mild hypertonic saline (2-5%) is widely used as an experimental model of pain in humans. Therefore, we tested whether TRPV4 participates in the transduction of hypertonic stimuli. Intradermal injection of 2% (607 mOsm) or 10% (3,250 mOsm) saline solution in the hind paw of rats induced a concentration-dependent pain-related behavior, flinching. Sensitization with prostaglandin E(2) (PGE(2)) caused a 7-fold increase in the number of flinches induced by 2% saline but failed to increase those caused by 10% saline. Spinal administration of antisense oligodeoxynucleotides to TRPV4 caused a 46% decrease in the number of flinches induced by 2% saline, but there was no change in flinching induced by 10% saline. Similarly, only the nociceptive behavior caused by 2% saline was reduced in TRPV4(-/-) knockout mice. The TRPV4-mediated nociceptive behaviors induced by hyper- and hypotonic stimuli were dependent on Src tyrosine kinase. We suggest TRPV4 is a transducer in primary afferents that mediates nociceptive behavior induced by small increases or decreases in osmolarity. Such changes in osmolarity might contribute to pain in inflammatory and neuropathic states. PMID- 16213086 TI - Neuropathic pain associated with non-surgical treatment of breast cancer. PMID- 16213087 TI - Relationship of pain and symptoms to pubertal development in adolescents. AB - Temporomandibular disorder (TMD) pain, abdominal pain, migraine and tension-type headache are more prevalent in women than in men. This study assessed the relationship of back pain, headache, abdominal pain, TMD pain, and the presence of multiple pain conditions to gender and pubertal development in a cross sectional, population-based survey of adolescents. We also examined the association between pubertal development and depressive and somatic symptoms, factors often associated with pain in adults. We hypothesized that prevalence of all pain conditions, as well as rates of other symptoms, would increase as puberty progresses in females, but not males. Subjects (3,101 boys and girls, 11 17 years old, selected from an HMO population) reported on the presence of each pain condition in the prior 3 months and completed scales assessing pubertal development, and depressive and somatic symptoms. Data were analyzed using descriptive statistics and multivariate logistic regression. Prevalence rates were weighted for factors affecting response. Prevalence of back pain, headache and TMD pain increased significantly (odds ratios, OR=1.4-2.0, P<0.001) and stomach pain increased marginally with increasing pubertal development in girls. Rates of somatization, depression and probability of experiencing multiple pains also increased with pubertal development in girls (P<0.0001). For boys, prevalence of back (OR=1.9, P<0.0001) and facial pain (OR=1.5, P<0.01) increased, stomach pain decreased somewhat and headache prevalence was virtually unchanged with increasing maturity. For both sexes, pubertal development was a better predictor of pain than was age. Thus it appears that pain, other somatic symptoms and depression increase systematically with pubertal development in girls. PMID- 16213088 TI - The feasibility of color Doppler ultrasonography for caudal epidural steroid injection. AB - Although it entails a radiation hazard risk, the use of fluoroscopy during caudal epidural steroid injection has increased to help place the medication more accurately and allowed physicians to maximize the procedure's therapeutic success rate. To investigate the feasibility of using real-time high resolution ultrasonography for guiding the epidural needle into the caudal epidural space and to confirm any vascular intake of medication, we performed color Doppler ultrasonography while medication was being injected into the caudal epidural space of 53 patients with low back pain and sciatica. We defined the injection as being successful if unidirectional flow (observed as one dominant color) of the solution was observed with color Doppler ultrasonography through the epidural space beneath the sacrococcygeal ligament, with no flows being observed in other directions (observed as multiple colors). The correct placement of the medication was then confirmed by fluoroscopy. In 52 of the 53 subjects, the medications were successfully injected into the caudal epidural space with ultrasonography assistance. In fluoroscopy, of these 52 patients, 50 revealed correct placement of the medicine into the epidural space. In conclusion, ultrasonography may be a reliable imaging modality for caudal epidural steroid injection, and its several advantages such as its convenience and the lack of a radiation hazard, make it preferable to fluoroscopy. PMID- 16213089 TI - A cannabinoid agonist, WIN 55,212-2, reduces neuropathic nociception induced by paclitaxel in rats. AB - Paclitaxel is an effective antineoplastic drug treatment used as an anti-tumoral therapy. Unfortunately its use is associated with unwanted side effects, which include the development of peripheral neuropathies and neuropathic pain, greatly affecting the quality of life of patients. It is well known that agonists of the cannabinoid receptor are able to reduce hyperalgesia and allodynia that develop after nerve injury. Our aim was to evaluate the efficacy of the cannabinoid agonist WIN 55,212-2 to reduce the thermal hyperalgesia and the tactile allodynia induced by administration of paclitaxel in rats. Present results demonstrate that WIN 55,212-2 (1 mg/kg i.p.) significantly reduced the heat (P<0.0001) and the mechanical (P=0.0003) withdrawal thresholds, the dose being smaller than that required to reach similar effects in the sciatic nerve constriction model (1.5 mg/kg). When the cannabinoid tetrad test was evaluated to measure behavioral modifications, it was found that WIN 55,212-2 (1mg/kg) did not induce changes either in body temperature or in immobility time, and only a reduction in spontaneous motility was recorded. This effect was antagonized by SR 141716A, suggesting the involvement of the CB1 receptor, although the participation of CB2 receptors cannot be excluded from this study. When WIN 55,212-2 was administered intraplantar, no differences were observed between the injected paw and the contralateral paw, suggesting that systemic mechanisms are needed to reach effectiveness. From these results we suggest that cannabinoids may be an interesting alternative to reduce neuropathic symptoms induced by paclitaxel, however more work is required to assess this possibility. PMID- 16213091 TI - Varicella zoster virus induces neuropathic changes in rat dorsal root ganglia and behavioral reflex sensitisation that is attenuated by gabapentin or sodium channel blocking drugs. AB - Reactivation of latent varicella zoster virus (VZV) within sensory trigeminal and dorsal root ganglia (DRG) neurons produces shingles (zoster), often accompanied by a chronic neuropathic pain state, post-herpetic neuralgia (PHN). PHN persists despite latency of the virus within human sensory ganglia and is often unresponsive to current analgesic or antiviral agents. To study the basis of varicella zoster-induced pain, we have utilised a recently developed model of chronic VZV infection in rodents. Immunohistochemical analysis of DRG following VZV infection showed the presence of a viral immediate early gene protein (IE62) co-expressed with markers of A- (neurofilament-200; NF-200) and C- (peripherin) afferent sensory neurons. There was increased expression of neuropeptide Y (NPY) in neurons co-expressing NF-200. In addition, there was an increased expression of alpha2delta1 calcium channel, Na(v)1.3 and Na(v)1.8 sodium channels, the neuropeptide galanin and the nerve injury marker, Activating Transcription Factor 3 (ATF-3) as determined by Western blotting in DRG of VZV-infected rats. VZV infection induced increased behavioral reflex responsiveness to both noxious thermal and mechanical stimuli ipsilateral to injection (lasting up to 10 weeks post-infection) that is mediated by spinal NMDA receptors. These changes were reversed by systemic administration of gabapentin or the sodium channel blockers, mexiletine and lamotrigine, but not by the non-steroidal anti-inflammatory agent, diclofenac. This is the first time that the profile of VZV infection-induced phenotypic changes in DRG has been shown in rodents and reveals that this profile appears to be broadly similar (but not identical) to changes in other neuropathic pain models. PMID- 16213092 TI - Multilevel somatosensory system disinhibition in children with migraine. AB - Although migraine is characterised by an abnormal cortical excitability level, whether the central nervous system is hyper- or hypo-excitable in migraine still remains an unsolved problem. The aim of our study was to compare the somatosensory evoked potential (SEP) recovery cycle, a marker of the somatosensory system's excitability, in a group of 15 children suffering from migraine without aura (MO) (mean age 11.7+/-1.6 years, five males, 10 females) and 10 control age-matched subjects (CS) (mean age 10.9+/-2.1 years, six males, four females). We calculated the SEP's latency and amplitude modifications after paired electrical stimuli at 5, 20 and 40 ms interstimulus intervals (ISIs), comparing it with a single stimulus condition assumed as the baseline. In MO patients, the amplitudes of the cervical N13 and of the cortical N20, P24 and N30 responses at 20 and 40 ms ISIs showed a higher recovery than in CS (two-way ANOVA, P<0.05). Since, the SEP recovery cycle depends on the inhibitory interneuron function, our findings suggest that a somatosensory system disinhibition takes place in migraine. This is a generalized phenomenon, not limited to the cerebral cortex, but concerning also the cervical grey matter. The SEP recovery cycle reflects the intracellular concentration of Na(+), therefore, the shortened recovery cycle in our MO patients suggests a high level of intracellular Na(+) and a consequent depolarized resting membrane potential, possibly due to an impaired Na(+) -K(+) ATPase function in migraine. PMID- 16213093 TI - Heart rate mediation of sex differences in pain tolerance in children. AB - Despite evidence supporting the existence of important sex-related differences in pain, the mechanisms underpinning such differences are not well understood. The aim of this study is to examine the relationship between sex and pubertal differences in autonomic arousal and pain tolerance to laboratory pain stimuli in healthy children. We tested the following specific hypotheses: (1) females would have greater autonomic arousal and less pain tolerance than males, and (2) this sex difference in pain tolerance would be mediated by autonomic arousal. Participants were 244 healthy children (50.8% female, mean age 12.73+/-2.98 years, range 8-18 years). Separate 4-trial blocks of cutaneous pressure and thermal pain stimuli were presented in counterbalanced order. Heart rate (HR) was recorded during 2-3 min periods preceding each block and a 1-min period between trials. Results indicated lower tolerance in females for cutaneous pressure, but not thermal pain, compared to males. In addition, pre-trial HR was greater for females than males. Mediation analyses suggested that sex differences in pressure pain tolerance were accounted for by sex differences in pre-trial HR. There were also significant effects for puberty, but these did not vary by sex. Overall, early pubertal children had greater pre-trial HR and less pain tolerance than those in late puberty for both cutaneous pressure and thermal pain across sex. These results suggest that autonomic arousal may be a mediator of sex-related differences in pain responses in children. PMID- 16213094 TI - Improving immunoassays based on the analysis of scanning force microscopy images. AB - Scanning force microscopy has been demonstrated to be an effective binding event detection step for immunoassays. In its simplest form--analysing small area images--the detection limit of the scanning force microscopic immunoassay (SFMIA) has been shown to be comparable to existing techniques. In the present work, we have examined how the performance of image analysis-based SFMIA can be improved. Firstly, we have used a surface analysis parameter that increases linearly with the concentration of binding events. This parameter--the surface area ratio--is the ratio of the surface area after antigen binding to the surface area of the original biospecific surface. With this parameter, SFMIA images can be rapidly analysed and converted into assay units. Secondly, we have demonstrated that by using silicon wafer supports that carry fiducial marks we can relocate to very high accuracy onto the biospecific surfaces and identify the changes due to antigen binding. By relocating in this manner the signal to noise ratio of the technique is enhanced. Thirdly, from simulations we have determined the SFM tip size and image area that optimizes the immunoassay sensitivity. PMID- 16213095 TI - East Coast fever and multiple El Nino Southern oscillation ranks. AB - East Coast fever (ECF), a tick-borne disease of cattle, is a major constraint to livestock development in Africa in general and southern Zambia in particular. Understanding the transmission patterns of this disease complex is very difficult as shown by previous studies in southern and eastern Zambia due to the interplay of risk factors. In this long-term study, we investigated whether global weather changes had any influence on disease transmission in traditionally kept cattle in southern Zambia. The results from this study show a strong association between increased Theileria parva contacts in cattle and the presence of El Nino, clearly linking a simple climatic index to disease outbreaks. We therefore propose that in southern Zambia, the simple and readily available multiple El Nino Southern oscillation index (MEI) ranks be used in planning ECF control programmes and early warning. PMID- 16213096 TI - Anthelmintic resistance of nematode parasites of small ruminants in eastern Ethiopia: exploitation of refugia to restore anthelmintic efficacy. AB - Faecal egg count reduction tests (FECRT) were conducted in May 2003 to determine the efficacy of anthelmintics used for treatment against nematode parasites in separately managed sheep and goat flocks at Alemaya University in eastern Ethiopia. These tests revealed high levels of anthelmintic resistance to albendazole, tetramisole, the combination of these two drugs, and to ivermectin in the goat flock (predominantly infected by Haemonchus contortus and Trichostrongylus spp.), whereas all drugs were highly efficacious in the sheep flock. A second FECRT confirmed these observations. Following this, a new management system was implemented on the goat flock for a period of 9 months (January-September 2004) in an attempt to restore the anthelmintic efficacy. This involved a combination of measures: eliminating the existing parasite infections in the goats, exclusion from the traditional goat pastures, and introducing communal grazing of the goats with the university sheep flock and livestock owned by neighbouring small-holder farmers. A second series of FECRTs (Tests 3 and 4) conducted 7 months after this change in management, showed high levels of efficacy to all three drugs (albendazole, tetramisole and ivermectin) in the goat flock. This is the first field study to demonstrate that anthelmintic efficacy in the control of nematode parasites of small ruminants can be restored by exploiting refugia. PMID- 16213097 TI - A survey on anthelmintic resistance in nematode parasites of sheep in the Slovak Republic. AB - The prevalence of anthelmintic resistance on 27 sheep farms in Slovakia was investigated in 2003 and 2004 using the faecal egg count reduction test (FECRT) according to the WAAVP guidelines. Resistance to albendazole was detected on one farm (3.7%) and suspected on two farms (7.4%) out of 27 sheep flocks. Resistance to ivermectin was tested on 26 farms. On six (23.1%) farms, results indicated the presence of ivermectin resistance. Resistance to ivermectin was suspected on eight farms (30.8%). However, it is also possible that generic ivermectin anthelmintics used in survey have a lower efficacy against sheep nematodes. PMID- 16213098 TI - Conformation and stability of elastase from Atlantic cod, Gadus morhua. AB - Metal binding and conformational stability characteristics of psychrophilic elastase (ACE) from Atlantic cod (Gadus morhua) has been investigated. Chelation to Ca(2+) was found to be important for maintaining the biologically active conformation and for the thermal stability of the enzyme. However, presence of metal ions such as Zn(2+), Fe(3+) and Cu(2+) was found to inhibit its hydrolytic activity and so did the chelating agent EDTA. Both pH and guanidinium chloride induced denaturation of the enzyme was followed by monitoring the changes in the tryptophan fluorescence. ACE exhibited a simple two-state unfolding pattern in both acidic and basic conditions with the midpoint of transition at pH values 4.08 and 10.29, respectively. Guanidinium chloride and heat induced denaturation of the enzyme was investigated at two pH values, 5.50 and 8.00, wherein the enzyme possesses similar tertiary structure but differ in its hydrolytic activity. Guanidinium chloride induced denaturation indicated that the enzyme unfolds with a C(m) of 1.53 M at pH 8.0 and a DeltaG(H2O) of 6.91 kJ mol(-1) (28.65 J mol(-1) residue(-1)) which is the lowest reported for psychrophilic enzymes investigated till-date. However, at pH 5.50, DeltaG(H2O) value is slightly lowered by 0.65 kJ mol(-1) consistent with the observed increase in the apparent quenching constant obtained with acrylamide. On the other hand, increase in T(m) by 38.45 degrees C was observed for the enzyme at acid pH (5.50) in comparison to the heat induced unfolding at pH 8.0. The increase in the apparent T(m) has been attributed to the possible weak intermolecular association of the enzyme molecules at moderately high temperatures that is favoured by the increase in the accessible surface area / dynamics under acidic conditions. The stability characteristics of ACE have been compared with the available data for mesophilic porcine pancreatic elastase and possible mechanism for the low temperature adaptation of ACE has been proposed. PMID- 16213099 TI - Expression of tcaA and mprF and glycopeptide resistance in clinical glycopeptide intermediate Staphylococcus aureus (GISA) and heteroGISA strains. AB - Two genes recently associated with glycopeptide intermediate resistance in Staphylococcus aureus (GISA) are mprF and tcaA, with inactivation causing shifts in vancomycin resistance. This study reveals that expression levels of both genes are similar in groups of clinical GISA, heteroGISA and glycopeptide susceptible strains, suggesting no association with clinical isolates. PMID- 16213100 TI - Histone-deacetylase inhibitors may accelerate the aging process in stem cell dependent mammals: stem cells, Ku70, and Drosophila at the crossroads. AB - The exact contribution of the sirtuin family of NAD+-dependent histone deacetylases to longevity in metazoans is, at present, not completely understood but nonetheless regarded as significant. Despite the rapidly accreting evidence solidifying the role of NAD+-dependent histone deacetylase activity in longevity promoting experimental interventions, the utility of histone-deacetylase inhibitors in the management of a diverse group of oncologic conditions draws question to the notion of universally beneficial effects of experimental interventions designed to promote deacetylase activity. The recent determination that overexpression of any one of the seven human sirtuin deacetylases fails to extend replicative lifespan in differentiated human cells calls attention to the possibility of unforeseen complexity in the determinants of human lifespan. Furthermore, inhibitors of histone deacetylases have been shown to actually increase lifespan in Drosophila. Delineation of the disparate effects of histone deacetylase activity in stem cells, progenitor cells, and fully differentiated cells may confirm initial findings suggesting that histone-deacetylase inhibitors push malignant cells towards terminal differentiation, while simultaneously exerting a proliferative and differentiating effect on normal stem cells. This effect may ultimately exert an accelerating influence on the aging of the stem cell population and consequently produce detrimental alterations in stem and progenitor cell populations that compromise organismal-level longevity in mammals, in contrast to findings in Drosophila. This opens the possibility of a new side effect to a widely used chemotherapeutic, as well as the possibility of the generation of novel experimental systems that could leverage the putatively pro-aging influence of histone-deacetylase inhibitors to explore aging. PMID- 16213101 TI - Polyarticular osteoarthritis--two major phenotypes hypothesized. AB - Osteoarthritis is the commonest form of arthritis, at least amongst Caucasians and is frequently polyarticular. Genetic factors are now considered pivotal in the aetiopathogenesis of polyarticular osteoarthritis (POA). This document proposes a nexus between the gene most commonly mutated amongst Caucasian peoples, notably the HFE gene and an appreciable subset of POA patients who have a clinically recognisable OA phenotype. It is hypothesised that there are at least 2 major POA phenotypes each of which is associated with discrete genotypes. Type 1 POA characterized by Heberden's or Bouchard's nodes with prominent DIP, PIP, knee joint (medial compartment) and Great toe MTP joint involvement corresponds to the putative nodal generalized form of OA or NGOA (proposed Type 1 POA phenotype). As yet no genetic marker has been defined for this POA subset. The second is a hitherto less well recognized phenotype characterized by involvement of the index and/or middle finger metacarpophalangeal (MCP2,3) joints and the elbows, ankles and possibly the intertarsal and tarsometatarsal joints. The hip and knee joints may sometimes also be involved. This different joint distribution corresponds closely to the pattern observed in the arthropathy that often accompanies hereditary haemochromatosis. It is predicted that mutations in the HFE gene will associate strongly with the proposed Type 2 POA phenotype and serve as a genetic marker for this clinically recognisable subset. PMID- 16213102 TI - Oxygen-induced reperfusion-injury is caused by ROS: Amelioration is possible by recombinant-DNA antioxidant enzymes and mimics in selected tissues. AB - Billions of years of photolytic cleavage of the water molecule has led to a build up at sea-level of a "toxic" oxygen concentration in the atmosphere (to 20%). Unfortunately, this dioxygen is likely to be converted in the mitochondria (cell organelles) during cellular respiration to generate reactive oxygen species (ROS); including free-radicals such as superoxide anion (.O2') and hydroxyl radical (.OH) (peroxide O2") a ROS is not a free radical because its electrons are paired). Development of recombinant-DNA improved isoenzyme forms (or mimics) of antioxidant enzymes such as superoxide dismutases is predicted in this hypothesis to be utilised to ameliorate reperfusion-injury (and other oxygen induced molecular pathology). Introduction into the human genome of the genes for expression of antioxidant enzymes, to order in particular tissues, is imminent. Furthermore, O2 itself will be recognised universally as a harmful gas that can subject the cell to oxidative stress; because it produces ROS, such as the superoxide anion when it acts as the terminal electron acceptor in cellular respiration in the production of water from hydrogen ions in mitochondria. In conclusion, therefore, oxygen-induced injury in humans can no longer be accepted during medical techniques such as reperfusion procedures, because this is associated with reperfusion-injury that can be the cause of several serious medical conditions arising from biomolecular pathology. Some of this is caused by isoforms of cytochromes P450 (CYP; EC 1.14.14.1) such as 3A4, 2D6 and 2C19. These can generate ROS in the liver at low substrate concentrations by futile recycling of oxygen. PMID- 16213103 TI - A prospective evaluation of the impact of 18-F-fluoro-deoxy-D-glucose positron emission tomography staging on survival for patients with locally advanced esophageal cancer. AB - PURPOSE: To determine the impact of 18-F-fluoro-deoxy-D-glucose positron emission tomography (FDG-PET) in the staging and prognosis of patients with locally advanced esophageal cancer (LAEC). METHODS AND MATERIALS: Between January 2000 and October 2004, all patients with LAEC evaluated in the Department of Radiation Oncology were considered for enrollment into a Phase II trial of preoperative chemoradiation. Entry required a staging whole-body FDG-PET scan. RESULTS: One hundred ten consecutive patients were evaluated; 38 were ineligible for reasons including treatment elsewhere, prior malignancy, or refusal of treatment. After conventional staging (clinical examination, endoscopic ultrasound, and chest/abdominal computerized tomography), 33 patients were ineligible because of metastatic disease or poor performance status. Of the remaining 39 patients, 23 were confirmed to have LAEC after FDG-PET staging and were treated in the Phase II trial (Cohort I). Sixteen patients, however, had FDG-PET findings consistent with occult metastatic disease and were deemed ineligible for the trial but were treated with curative intent (Cohort II). The 2-year survival rate for the 23 patients in Cohort I was 64%, compared with 17% (p=0.003) for patients in Cohort II (FDG-PET positive). CONCLUSIONS: More than one-third of patients determined to have LAEC with conventional staging were upstaged with the use of FDG-PET. Despite comparable therapy, upstaging with FDG-PET predicts poor 2-year survival. PMID- 16213104 TI - Long-term outcome of concurrent chemotherapy and reirradiation for recurrent and second primary head-and-neck squamous cell carcinoma. AB - PURPOSE: To define favorable pretreatment characteristics for overall survival (OS), progression-free survival (PFS), locoregional control, and freedom from distant metastasis for patients with recurrent and second primary head-and-neck cancer treated with concomitant chemotherapy and reirradiation. METHODS AND MATERIALS: Our study population comprised a subset of 115 previously irradiated patients without overt metastases from 304 poor-prognosis head-and-neck cancer patients treated in seven consecutive phase I-II protocols. Of the 115 patients, 49, who had undergone surgical resection, were treated with a median of four cycles of concurrent chemotherapy and reirradiation and 66, who had not undergone surgical resection, were treated with a median of five cycles. The following regimens were used: 5-fluorouracil and hydroxyurea concurrent with reirradiation (FHX) (n=14), cisplatin plus FHX (n=23), paclitaxel plus FHX (n=42), gemcitabine plus paclitaxel and 5-fluorouracil concurrent with reirradiation (n=26), and irinotecan plus FHX (n=10). RESULTS: The median lifetime radiation dose was 131 Gy. The median follow-up for surviving patients was 67.4 months (range, 18.5 158.7). The median OS and PFS was 11 and 7 months (range, 0.2-158.7), respectively. The 3-year OS, PFS, locoregional control, and freedom from distant metastasis rate was 22%, 33%, 51%, and 61%, respectively. Multivariate analysis identified reirradiation dose, triple agent (cisplatin-, paclitaxel-, or gemcitabine-containing chemotherapy), and surgery before protocol treatment as independently prognostic for OS, PFS, and locoregional control. Triple-agent chemotherapy was prognostic for freedom from distant metastasis. Nineteen patients died of treatment-related toxicity, five of these of carotid hemorrhage. CONCLUSION: For recurrent and second primary head-and-neck cancer, trimodality therapy with surgery, concurrent chemotherapy, and reirradiation for a full second dose offers potential for long-term survival. Owing to the substantial toxicity and lack of an optimal regimen, reirradiation of recurrent head-and-neck cancer should be limited to clinical trials. PMID- 16213105 TI - Preliminary results of radiation dose escalation for locally advanced nasopharyngeal carcinoma. AB - PURPOSE: To study the safety and efficacy of dose escalation in tumor for locally advanced nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: From September 2000 to June 2004, 50 patients with T3-T4 NPC were treated with intensity modulated radiotherapy (IMRT). Fourteen patients had Stage III and 36 patients had Stage IVA-IVB disease. The prescribed dose was 76 Gy to gross tumor volume (GTV), 70 Gy to planning target volume (PTV), and 72 Gy to enlarged neck nodes (GTVn). All doses were given in 35 fractions over 7 weeks. Thirty-four patients also had concurrent cisplatin and induction or adjuvant PF (cisplatin and 5 fluorouracil). RESULTS: The average mean dose achieved in GTV, GTVn, and PTV were 79.5 Gy, 75.3 Gy, and 74.6 Gy, respectively. The median follow-up was 25 months, with 4 recurrences: 2 locoregional and 2 distant failures. All patients with recurrence had IMRT alone without chemotherapy. The 2-year locoregional control rate, distant metastases-free and disease-free survivals were 95.7%, 94.2%, and 93.1%, respectively. One treatment-related death caused by adjuvant chemotherapy occurred. The 2-year overall survival was 92.1%. CONCLUSIONS: Dose escalation to 76 Gy in tumor is feasible with T3-T4 NPC and can be combined with chemotherapy. Initial results showed good local control and survival. PMID- 16213106 TI - COX-2 overexpression in pretreatment biopsies predicts response of rectal cancers to neoadjuvant radiochemotherapy. AB - PURPOSE: To determine the utility of COX-2 expression as a response predictor for patients with rectal cancer who are undergoing neoadjuvant radiochemotherapy (RCT). METHODS AND MATERIALS: Pretreatment biopsies (PTB) from 49 patients who underwent RCT were included. COX-2 and proliferation in PTB were assessed by immunohistochemistry (IHC) and apoptosis was detected by TUNEL stain. Response to treatment was assessed by a 5-point tumor-regression grade (TRG) based on the ratio of residual tumor to fibrosis. RESULTS: Good response (TRG 1+2), moderate response (TRG 3), and poor response (TRG 4+5) were seen in 21 patients (42%), 11 patients (22%), and 17 patients (34%), respectively. Patients with COX-2 overexpression in PTB were more likely to demonstrate moderate or poor response (TRG 3+4) to treatment than were those with normal COX-2 expression (p=0.026, chi square test). Similarly, poor response was more likely if patients had low levels of spontaneous apoptosis in PTBs (p=0.0007, chi-square test). CONCLUSIONS: COX-2 overexpression and reduced apoptosis in PTB can predict poor response of rectal cancer to RCT. As COX-2 inhibitors are commercially available, their administration to patients who overexpress COX-2 warrants assessment in clinical trials in an attempt to increase overall response rates. PMID- 16213107 TI - Oxygen-modifying treatment with ARCON reduces the prognostic significance of hemoglobin in squamous cell carcinoma of the head and neck. AB - PURPOSE: To evaluate the prognostic significance of hemoglobin (Hb) levels measured before and during treatment with accelerated radiotherapy with carbogen and nicotinamide (ARCON). METHODS AND MATERIALS: Two hundred fifteen patients with locally advanced tumors of the head and neck were included in a phase II trial of ARCON. This treatment regimen combines accelerated radiotherapy for reduction of repopulation with carbogen breathing and nicotinamide to reduce hypoxia. In these patients, Hb levels were measured before, during, and after radiotherapy. RESULTS: Preirradiation and postirradiation Hb levels were available for 206 and 195 patients respectively. Hb levels below normal were most frequently seen among patients with T4 (p < 0.001) and N2 (p < 0.01) disease. Patients with a larynx tumor had significantly higher Hb levels (p < 0.01) than other tumor sites. During radiotherapy, 69 patients experienced a decrease in Hb level. In a multivariate analysis there was no prognostic impact of Hb level on locoregional control, disease-free survival, and overall survival. Primary tumor site was independently prognostic for locoregional control (p = 0.018), and gender was the only prognostic factor for disease-free and overall survival (p < 0.05). High locoregional control rates were obtained for tumors of the larynx (77%) and oropharynx (72%). CONCLUSION: Hemoglobin level was not found to be of prognostic significance for outcome in patients with squamous cell carcinoma of the head and neck after oxygen-modifying treatment with ARCON. PMID- 16213108 TI - A clinical staging system and treatment guidelines for maxillary osteoradionecrosis in irradiated nasopharyngeal carcinoma patients. AB - PURPOSE: To develop a clinical staging system for maxillary osteoradionecrosis (ORN) in irradiated nasopharyngeal carcinoma (NPC) patients. METHODS AND MATERIALS: The data of maxillary ORN cases among 1,758 irradiated NPC patients were analyzed. A staging system based on the degrees of bone exposure (E), infection (I), and bleeding (B) was developed. Correlations between various clinical parameters and stages of maxillary ORN and relationships between treatment modalities and outcomes at each stage were evaluated. Cumulative success of treatment and risk factors that affect treatment outcomes were analyzed. RESULTS: The incidence of maxillary ORN was 2.7% (48/1,758). TNM stage of NPC (p < 0.001), radiation dose (p = 0.029), and tooth extraction (p < 0.001) appeared to have significant influences on disease severity. Success rates between conservative therapy and surgical treatment were not significantly different for Stage I ORN but differed significantly for Stage II (p = 0.013) and Stage III (p = 0.008) lesions. Grade 3 infection and bleeding significantly jeopardized treatment success (p = 0.043 and 0.015, respectively). The risk ratios of treatment failure for Grade 3 infection and bleeding were 2.523 (p = 0.034) and 3.141 (p = 0.027), respectively. CONCLUSIONS: More serious maxillary ORN tended to occur in cases with more advanced NPC, higher radiation dose, and history of tooth extraction. Surgical treatment was usually required in Stage II and III ORN. The grades of infection and bleeding are important factors in guidance of treatment and prediction of outcomes. PMID- 16213109 TI - Response to cocaine, alone and in combination with methylphenidate, in cocaine abusers with ADHD. AB - Attention deficit hyperactivity disorder (ADHD) is prevalent in adult cocaine abusers. Yet, it remains to be determined how the response to cocaine differs in cocaine abusers with ADHD compared to cocaine abusers without ADHD. Further, since ADHD is commonly treated with stimulants, such as methylphenidate (MPH), it is important to examine whether MPH maintenance alters the response to cocaine in cocaine abusers with ADHD. Thus, the first phase of this study compared the response to cocaine in adult cocaine abusers with ADHD to those without ADHD. The second phase assessed the effects of oral sustained-release methylphenidate (MPH SR) maintenance (40 and 60 mg) on the response to cocaine only in those with ADHD. Cocaine abusers with ADHD (N=7) and without ADHD (N=7) who were not seeking treatment remained inpatient initially for 1 week, when the effects of cocaine alone were tested (Phase 1). Cocaine abusers with ADHD remained inpatient for an additional 3 weeks, during which the effects of cocaine during oral MPH-SR maintenance were tested (Phase 2). During cocaine fixed dosing sessions, participants received four injections of i.v. cocaine (0, 16 or 48 mg/70 kg), spaced 14 min apart. During cocaine choice sessions, participants had a choice between receiving i.v. cocaine (16 or 48 mg/70 kg) or two tokens, each exchangeable for 2 US dollars. Subjective effects related to ADHD symptoms (e.g. ratings of "Able to Concentrate") were significantly lower in cocaine abusers with ADHD compared to those without ADHD when placebo cocaine was administered. Active cocaine produced similar increases in cardiovascular and positive subjective effects in both groups and there was no difference in cocaine choice between the two groups. These data suggest that the response to cocaine is not different between cocaine abusers with ADHD compared to those without ADHD. When the cocaine abusers with ADHD were maintained on MPH-SR, cardiovascular effects were increased, however, this did not warrant termination of any test session. Maintenance on MPH-SR decreased some of the positive subjective effects of cocaine. Further, maintenance on a high dose of MPH-SR decreased cocaine choice. Thus, oral MPH-SR is safe in combination with repeated cocaine doses and decreases some of the positive and reinforcing effects of cocaine in cocaine abusers with ADHD. PMID- 16213110 TI - Role of the increased noradrenergic neurotransmission in drug self administration. AB - Psychostimulants increase extracellular monoamine concentrations in the CNS. While the contributions of dopamine (DA) and serotonin (5-HT) to the reinforcing effect of psychostimulants have been examined, less is known about the involvement of norepinephrine (NE). In the present study, cocaine, desipramine (DMI) and JZ-III-84 were made available to rhesus monkeys (n=4) responding under a progressive-ratio (PR) schedule. These compounds vary in their in vitro selectivities for blocking NE uptake relative to DA from high (DMI) to modest (JZ III-84) to non-selective (cocaine). Additionally, cocaine mixed with DMI in mg/kg dose-ratios of 1:1 to 1:3 was made available for self-administration. NE uptake inhibition by the mixture of cocaine and DMI at a ratio of 1:3 was evaluated in an ex vivo uptake assay. Cocaine (0.01-0.1 mg/(kg injection)) and JZ-III-84 (0.001-0.1 mg/(kg injection)) functioned as positive reinforcers with sigmoidal or biphasic dose-response functions, whereas DMI failed to do so. The addition of DMI to cocaine did not systemically alter self-administration of cocaine. In the ex vivo uptake assay, the addition of DMI to cocaine significantly increased the NE uptake inhibition compared to cocaine. These results support the conclusion that CNS NE is not involved in the reinforcing mechanism of psychostimulants. PMID- 16213111 TI - On the coevolution of genes and genetic code. AB - The canonical genetic code acts efficiently in minimizing the effects of mistranslations and point mutations. In the work presented we have also considered the effects of single nucleotide insertions and deletions on the optimality of the genetic code. Our results suggest that the canonical genetic code compensates for the ins/del mutations as well as mistranslations and point mutations. On the other hand, we highlighted the point that ins/del mutations have a lesser impact on the selected genes of Saccharomyces cerevisiae compared to randomly generated ones. We hypothesized that the codon usage preferences in S. cerevisiae genes are responsible for the higher efficiency of translation machinery in this organism. Our results support the conjecture that codon usage preferences render the genetic code more effective in minimizing the effects of ins/del mutations. PMID- 16213112 TI - Regulation of translation via mRNA structure in prokaryotes and eukaryotes. AB - The mechanism of initiation of translation differs between prokaryotes and eukaryotes, and the strategies used for regulation differ accordingly. Translation in prokaryotes is usually regulated by blocking access to the initiation site. This is accomplished via base-paired structures (within the mRNA itself, or between the mRNA and a small trans-acting RNA) or via mRNA-binding proteins. Classic examples of each mechanism are described. The polycistronic structure of mRNAs is an important aspect of translational control in prokaryotes, but polycistronic mRNAs are not usable (and usually not produced) in eukaryotes. Four structural elements in eukaryotic mRNAs are important for regulating translation: (i) the m7G cap; (ii) sequences flanking the AUG start codon; (iii) the position of the AUG codon relative to the 5' end of the mRNA; and (iv) secondary structure within the mRNA leader sequence. The scanning model provides a framework for understanding these effects. The scanning mechanism also explains how small open reading frames near the 5' end of the mRNA can down regulate translation. This constraint is sometimes abrogated by changing the structure of the mRNA, sometimes with clinical consequences. Examples are described. Some mistaken ideas about regulation of translation that have found their way into textbooks are pointed out and corrected. PMID- 16213113 TI - Amplification and sequencing of Brachyspira spp. specific portions of nox using paraffin-embedded tissue samples from clinical colitis in Austrian pigs shows frequent solitary presence of Brachyspiramurdochii. AB - Brachyspira infections are significant causes of enterocolitis in pigs. In order to differentiate pathogenic species (Brachyspira (Br.) hyodysenteriae, Brachyspira pilosicoli) from less pathogenic or non-pathogenic species (Brachyspira intermedia, Brachyspira innocens, Brachyspira murdochii) in paraffin embedded tissue samples a polymerase chain reaction (PCR) protocol allowing identification of Brachyspira at species level in archival material was developed. This approach was complemented by sequencing of the PCR amplification products. All seven cases presented with clinical and morphological Brachyspira associated enterocolitis. Br. hyodysenteriae was not identified in any of the cases, while Br. pilosicoli was identified in a single case in conjunction with Br. murdochii. One case each was found positive for Br. innocens and Br. intermedia. Interestingly, the majority of cases presented as single or double infections with Br. murdochii. In some of the pigs other pathogens, like porcine circovirus-2 or Lawsonia intracellularis were present. These observations point at the possibility that under certain conditions even Brachyspira species of low pathogenicity can multiplicate extensively and lead to Brachyspira-associated enterocolitis. PMID- 16213114 TI - Available medical choices for the management of menopause. AB - The age at which menopause occurs is a critical factor in the magnitude of its consequences. Most of the medium-to-long-term effects of oestrogen deprivation depend on their duration. The timing of the last menstruation is therefore important, but hypoestrogenic amenorrhoea during the reproductive age is also a relevant factor in the evaluation of individual risks. In recent years, moving post-menopausal women from the lowest point of ovarian hypofunction has been the most important motivation for developing guidelines for the hormonal management of menopause. However, recent data suggest that this may be associated with an unacceptable increase in morbidity in a number of women. Concerns about long-term hormone replacement therapy (HRT) at menopause have recently enhanced interest in a group of molecules that act on the oestrogen receptor with selective effects, known as selective oestrogen receptor modulators (SERMs). Of these, Raloxifene has been approved for the treatment and prevention of osteoporosis, and exhibits a pattern of actions particularly well matched to the needs and concerns of post menopausal women. Further studies on SERMs may open up new vistas in patient specific management of post-menopausal health. Finally, debates on the specific health consequences of menopause deal mainly with the risk of chronic disease. Gynaecologists and other health professionals would be advised to develop intervention strategies at menopause according to the continuum of a woman's life, beginning at the post-menarche and extending into later life. PMID- 16213116 TI - Shifting the onset of a moving sound source: a Frohlich effect in spatial hearing. AB - When observers are presented with a visual target in motion, they typically remember its onset position to be displaced in the direction of motion. The present study investigated a similar effect in the auditory modality. In a dark anechoic environment, an auditory target (short noise pulses) appeared randomly at a peripheral or a central azimuthal position and moved from left to right or from right to left along the frontal horizontal plane. Relative judgments were made to determine the onset position of motion: Employing a two-alternative forced-choice task, listeners compared the onset position of the target to a 2-s visual reference stimulus presented at the left or right of the auditory onset position. In comparison with stationary targets, the onset positions of moving targets were localized as displaced in the direction of motion. The most prominent displacement occurred when the visual reference stimulus was presented after the auditory motion. With the reference stimulus presented before the auditory motion, the displacement was significantly reduced. Moreover, the displacement was stronger with peripheral than with central onset positions. These findings suggest the existence of a potential analogue of the Frohlich effect in the auditory modality. An auditory spatial attention mechanism is proposed that may have given rise to the observed pattern of results. PMID- 16213115 TI - Comparison of possible carcinogenic estradiol metabolites: effects on proliferation, apoptosis and metastasis of human breast cancer cells. AB - OBJECTIVES: Certain estradiol metabolites may play a pivotal role in breast carcinogenesis. Of special interest are the metabolites 2-hydroxyestradiol (2 OHE2), which can react anti-carcinogenically, and 4-hydroxyestradiol (4-OHE1) and 16a-hydroxyestrone (16-OHE1), which may have procarcinogenic potential. In the present study, we have compared for the first time the effect of these metabolites and their parent substance 17beta-estradiol (E2) on proliferation, apoptosis, apoptosis markers and markers of metastatic property of human breast cancer cells. METHODS: MCF-7 cells (human estrogen-receptor positive metastatic breast cancer cell line) were incubated with the estrogens at concentrations of 0.1-100 nM. Cell proliferation rate was measured by the ATP-assay. Apoptosis was measured by cell death assay and the apoptosis markers cytochrome C, Bcl-2, Fasl and p53 were determined in cell lysates by ELISAs. The markers of metastatic property of the cell line, VEGF and MCP-1 were measured in the cell supernatant by ELISAs. RESULTS: The estrogens E2, 4-OHE2 and 16-OHE1 display a proliferative effect on MCF-7 cells which is accompanied by a down-regulation of apoptosis. Various markers of apoptosis such as Bcl-2, cytochrome C and p53 appear to be involved. No significant effect was found for the metabolite 2-OHE2. VEGF and MCP 1 were up-regulated by E2 and 16-OHE1, whereas 2-OHE2 and 4-OHE2 did not show any effect. CONCLUSIONS: The most potent estrogen regarding proliferation, apoptosis and metastasis of breast cancer cells seems to be estradiol. However, the estradiol metabolites 4-OHE2 and 16-OHE1 elicit similar properties on cell proliferation, apoptosis and metastasis as compared to estradiol but only at higher concentrations. In contrast 2-OHE2 did not show any significant effect on these parameters. Thus, intracellular estradiol metabolism may determine an individual's risk for breast carcinogenesis. PMID- 16213117 TI - Enhancement of peritoneal macrophage phagocytic activity against Leishmania major by garlic (Allium sativum) treatment. AB - It has been shown that garlic (Allium sativum) extract modulates immune responses. Macrophage activation is required to establish control of the intracellular infection and progressive disease of leishmaniasis. In this study, we consider the effect of a garlic extract and a fraction isolated from it on the engulfment and destruction of Leishmania major by resident peritoneal macrophages of Balb/c mice. In regard to this study, the infiltration of macrophages in the peritoneal cavity after garlic treatment and the rate of amastigotes per macrophage were determined. The results show that a single dose of 20 mg/kg garlic extract intraperitoneally (i.p.) alters the number of peritoneal macrophages for at least 2 weeks. Intraperitoneal injection of garlic extract (20 mg/kg) or its protein fraction (0.04 mg/kg) augments parasite engulfment and destruction of intracellular amastigotes by macrophages. PMID- 16213118 TI - Anti-inflammatory effect of Houttuynia cordata injection. AB - Houttuynia cordata (Saururaceae) injection (HCI) is a traditional Chinese medicine used in China. It was chosen as one of eight types of traditional Chinese medicine that play a unique role in severe acute respiratory syndrome (SARS) owing to the effect of curbing inflammation. In order to validate this plausible anti-inflammatory property, the chemical composition of HCI has been analysed by GC/MS, 22 components were identified, and the inflammation induced by carrageenan in the rat pleurisy model and by xylene in the mice ear edema model was adopted to study the anti-inflammatory activity of HCI. Injection of carrageenan into the pleural cavity elicited an acute inflammatory response characterized by protein rich fluid accumulation and leukocyte infiltration in the pleural cavity. The peak inflammatory response was obtained at 24 h when the fluid volume, protein concentration, C-reactive protein and cell infiltration were maximums. The results showed that these parameters were attenuated by HCI at any dose and touched bottom at dose of 0.54 ml/100 g, although less strong than dexamethasone. This drug was also effective in inhibiting xylene induced ear edema, and the percentage of inhibition came to 50% at dose of 80 microl/20 g. The results clearly indicate that HCI have anti-inflammatory activity. PMID- 16213119 TI - Anti-diabetic effect of cinnamon extract on blood glucose in db/db mice. AB - The anti-diabetic effect of Cinnamomi cassiae extract (Cinnamon bark: Lauraceae) in a type II diabetic animal model (C57BIKsj db/db) was studied. Cinnamon extract was administered at different dosages (50, 100, 150 and 200 mg/kg) for 6 weeks. It was found that blood glucose concentration is significantly decreased in a dose-dependent manner (P<0.001) with the most in the 200 mg/kg group compared with the control. In addition, serum insulin levels and HDL-cholesterol levels were significantly higher (P<0.01) and the concentration of triglyceride, total cholesterol and intestinal alpha-glycosidase activity were significantly lower after 6 weeks of the administration. These results suggest that cinnamon extract has a regulatory role in blood glucose level and lipids and it may also exert a blood glucose-suppressing effect by improving insulin sensitivity or slowing absorption of carbohydrates in the small intestine. PMID- 16213120 TI - Hepatoprotective effects of rubiadin, a major constituent of Rubia cordifolia Linn. AB - The hepatoprotective effects of rubiadin, a major constituent isolated from Rubia cordifolia Linn., were evaluated against carbon tetrachloride (CCl4)-induced hepatic damage in rats. Rubiadin at a dose of 50, 100 and 200 mg/kg was administered orally once daily for 14 days. The substantially elevated serum enzymatic activities of serum glutamic oxaloacetic transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (SALP) and gamma-glutamyltransferase (gamma-GT) due to carbon tetrachloride treatment were dose dependently restored towards normalization. Meanwhile, the decreased activities of glutathione S-transferase and glutathione reductase were also restored towards normalization. In addition, rubiadin also significantly prevented the elevation of hepatic malondialdehyde formation and depletion of reduced glutathione content in the liver of CCl4 intoxicated rats in a dose dependent manner. Silymarin used as standard reference also exhibited significant hepatoprotective activity on post treatment against carbon tetrachloride induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that rubiadin has a potent hepatoprotective action against carbon tetrachloride induced hepatic damage in rats. PMID- 16213121 TI - Antiplasmodial diterpenoid from the leaves of Hyptis suaveolens. AB - Bioactivity-guided fractionation of the petroleum ether extract of the leaves of Hyptis suaveolens, widely used in Traditional Medicine, has led to the isolation of an abietane-type diterpenoid endoperoxide, 13 alpha-epi-dioxiabiet-8(14)-en-18 ol, displaying high antiplasmodial activity (IC(50) 0.1 microg/ml). PMID- 16213122 TI - Quinolinic acid reduces the antioxidant defenses in cerebral cortex of young rats. AB - Quinolinic acid (QA), the major metabolite of the kynurenine pathway, is found at increased concentrations in brain of patients affected by various common neurodegenerative diseases, including Huntington's disease and Alzheimer's disease. Recently, a role for QA in the pathophysiology of glutaric acidemia type I (GAI) was postulated. Considering that oxidative stress has been recently involved in the pathophysiology of the brain injury in these neurodegenerative disorders; in the present study, we investigated the in vitro effect of QA on various parameters of oxidative stress, namely total radical-trapping antioxidant potential (TRAP), total antioxidant reactivity (TAR), glutathione (GSH) levels, thiobarbituric acid-reactive substances (TBA-RS) measurement and chemiluminescence in cerebral cortex of 30-day-old rats. QA diminished the brain non-enzymatic antioxidant defenses, as determined by the reduced levels of TRAP, TAR and GSH. We also observed that QA significantly increased TBA-RS and chemiluminescence. Therefore, in vitro QA-treatment of rat cortical supernatants induced oxidative stress by reducing the tissue antioxidant defenses and increasing lipid oxidative damage, probably as a result of free radical generation. In addition, we examined the effect of QA on TBA-RS levels in the presence of glutaric acid (GA) and 3-hydroxyglutaric acid (3HGA), which are accumulated in GAI, as well as in the presence of 3-hydroxykynurenine (3HK), a tryptophan metabolite of the kynurenine pathway with antioxidant properties. It was verified that the single addition of QA or GA plus 3HGA to the incubation medium significantly stimulated in vitro lipid peroxidation. Furthermore, 3HK completely prevented the TBA-RS increase caused by the simultaneous addition of QA, GA and 3HGA. Taken together, it may be presumed that QA induces oxidative stress in the brain, which may be associated, at least in part, with the pathophysiology of central nervous system abnormalities of neurodegenerative diseases in which QA accumulates. PMID- 16213123 TI - Mancozeb exposure in vivo impairs mouse oocyte fertilizability. AB - Mancozeb is known to alter reproductive performance in exposed animals, but its specific mechanism of action is still unclear. We investigated whether in female mice of the F1 generation, mancozeb could affect oocyte ability to undergo complete meiotic maturation and fertilization. Female mice were treated with 50 and 500 mg/kg of mancozeb (or vehicle in the controls) from gestational day 2 to postnatal day 20. Results demonstrated that only at the highest dose, mancozeb induced a significant decrease in the number of ovulated eggs. Moreover, at this dose mancozeb caused a significant decrease of fertilizability related to a reduction of the formation of male and female pronuclei. PMID- 16213124 TI - Frequency of isolation and antimicrobial susceptibility of bacteria isolated from bloodstream infections at Children's Medical Center, Tehran, Iran, 1996-2000. AB - Antimicrobial susceptibility patterns of major bloodstream pathogens from Iran provide essential information regarding the selection of antibiotic therapy for patients with bloodstream infections (BSIs) living in Iran. Unfortunately, data regarding the isolation frequency and antimicrobial susceptibility patterns of endemic BSI pathogens are scarce in Iran. To shed some light on the susceptibility patterns of BSI pathogens endemic to Tehran, Iran, we investigated the antimicrobial susceptibility patterns of 2248 bloodstream isolates from patients in Children's Medical Center (CMC) Hospital in Tehran between January 1996 and December 2000. Microbiology reports of 24600 blood specimens collected from inpatients in CMC Hospital were retrospectively reviewed. Specimen culture, bacterial identification and disk diffusion susceptibility testing were performed according to National Committee for Clinical Laboratory Standards guidelines. Overall, Gram-positive bacteria comprised 72% (1627/2248) of recovered isolates and Gram-negative bacteria comprised 28% (621/2248). Coagulase-negative staphylococci (CoNS) comprised 48.4% of all isolates, followed by Staphylococcus aureus (16.7%) and Klebsiella spp. (8.5%). Among the 621 Gram-negative organisms, Klebsiella spp. (31%) were the most frequently isolated, followed by Escherichia coli (21%) and Pseudomonas aeruginosa (17%). The rates of oxacillin resistance for S. aureus and CoNS isolates were similar (60% versus 61%); however, the rate of S. aureus vancomycin resistance was almost twice that of CoNS resistance (21% versus 11%). Over 55% of S. pneumoniae were resistant to penicillin and co trimoxazole. Although all isolates of enterococci were susceptible to vancomycin, only 21% were susceptible to gentamicin. Among Gram-negative isolates, amikacin was shown to be very effective, with susceptibility rates of 84%. The susceptibility of Klebsiella spp. to ampicillin and co-trimoxazole was 1% and 39%, respectively. The susceptibility of Klebsiella spp., E. coli and Enterobacter spp. to ceftriaxone was 47%, 86% and 67%, respectively. There were notable differences in the order of the five most common organisms isolated from blood cultures, which can help set priorities for focused control efforts. Our findings highlight the importance of a nationwide surveillance programme to monitor the trends in isolation frequency of bacteria and their antimicrobial resistance patterns throughout Iran. PMID- 16213125 TI - Decreased agonist-stimulated mitochondrial ATP production caused by a pathological reduction in endoplasmic reticulum calcium content in human complex I deficiency. AB - Although a large number of mutations causing malfunction of complex I (NADH:ubiquinone oxidoreductase) of the OXPHOS system is now known, their cell biological consequences remain obscure. We previously showed that the bradykinin (Bk)-induced increase in mitochondrial [ATP] ([ATP](M)) is significantly reduced in primary skin fibroblasts from a patient with an isolated complex I deficiency. The present work addresses the mechanism(s) underlying this impaired response. Luminometry of fibroblasts from 6 healthy subjects and 14 genetically characterized patients expressing mitochondria targeted luciferase revealed that the Bk-induced increase in [ATP](M) was significantly, but to a variable degree, decreased in 10 patients. The same variation was observed for the increases in mitochondrial [Ca(2+)] ([Ca(2+)](M)), measured with mitochondria targeted aequorin, and cytosolic [Ca(2+)] ([Ca(2+)](C)), measured with fura-2, and for the Ca(2+) content of the endoplasmic reticulum (ER), calculated from the increase in [Ca(2+)](C) evoked by thapsigargin, an inhibitor of the ER Ca(2+) ATPase. Regression analysis revealed that the increase in [ATP](M) was directly proportional to the increases in [Ca(2+)](C) and [Ca(2+)](M) and to the ER Ca(2+) content. Our findings provide evidence that a pathological reduction in ER Ca(2+) content is the direct cause of the impaired Bk-induced increase in [ATP](M) in human complex I deficiency. PMID- 16213126 TI - Dorsolateral prefrontal cortex contribution to abnormalities of the P300 component of the event-related potential in schizophrenia. AB - Abnormalities of the P300 component of the event-related potential are a common finding in schizophrenia. It seems possible that the dysfunction in the dorsolateral prefrontal (DLPF) region that has been reported in schizophrenia contributes to this finding. To explore this possibility, we calculated the relationship between, on the one hand, P300 latency and amplitude and, on the other hand, the degree of DLPF atrophy (as measured by magnetic resonance imaging) and metabolic activity during an attentional task (as measured by positron emission tomography). Seventeen schizophrenia patients with a brief duration of illness and minimal exposition to treatment and 25 healthy controls were studied. Patients exhibited significantly lower metabolic activity in the DLPF region, but they did not show cortical atrophy. P300 amplitude was also significantly reduced in the schizophrenia patients compared with the controls. Right DLPF region metabolic activity correlated significantly with P300 amplitude. This pattern remained after partialling out the influence of activity in the hippocampus, superior temporal gyrus and parietal lobe. It is therefore suggested that the prefrontal cortex could be implicated in the P300 amplitude reduction in schizophrenia. PMID- 16213127 TI - In situ gelling, bioadhesive nasal inserts for extended drug delivery: in vitro characterization of a new nasal dosage form. AB - The purpose of this study was the preparation and characterization of sponge like, in situ gelling inserts based on bioadhesive polymers. Hydrophilic polymers (carrageenan, Carbopol, chitosan, hydroxypropyl methylcellulose (HPMC) K15M and E5, sodium alginate, sodium carboxy methylcellulose (NaCMC), polyvinyl pyrrolidone (PVP) 90, xanthan gum) were dissolved with/without the model drug oxymetazoline HCl in demineralized water and lyophilized into small inserts. The drug release, water uptake, mechanical properties, X-ray diffraction and bioadhesion potential of the nasal inserts were investigated. A sponge-like structure of nasal inserts was formed with amorphous, but not with crystalline polymers during the freeze-drying process. The insert hardness increased with the glass transition temperature of the polymer (PVP25 or = 17 mm. After 36 h, oocytes were retrieved and IVF was performed. Embryo transfer was done at the 4-8 cell embryo stage. Luteal support was initiated from the day of embryo transfer and continued for up to 14 days. Patients were randomised to luteal supplementation with either intravaginal micronised progesterone 200 mg three times daily (n=351) or oral dydrogesterone 10 mg twice daily (n=79). In cases of a positive pregnancy test, luteal support was continued for 12 weeks. Both dydrogesterone and micronised progesterone were associated with similar rates of successful pregnancies. Vaginal discharge or irritation were reported by 10.5% of patients given micronised progesterone. Significantly (p<0.05), more patients given dydrogesterone than micronised progesterone were satisfied with the tolerability of their treatment. There were no differences between the treatments with regard to liver function tests. PMID- 16213137 TI - Effects of storage conditions of Moringa oleifera seeds on its performance in coagulation. AB - Moringa oleifera is a plant whose seeds have coagulation properties for treating water and wastewater. In this study the coagulation efficiency of Moringa oleifera kept in different storage conditions were studied. The Moringa oleifera seeds were stored at different conditions and durations; open container and closed container at room temperature (28 degrees C) and refrigerator (3 degrees C) for durations of 1, 3 and 5 months. Comparison between turbidity removal efficiency of Moringa oleifera kept in refrigerator and room temperature revealed that there was no significant difference between them. The Moringa oleifera kept in refrigerator and room temperature for one month showed higher turbidity removal efficiency, compared to those kept for 3 and 5 months, at both containers. The coagulation efficiency of Moringa oleifera was found to be dependent on initial turbidity of water samples. Highest turbidity removals were obtained for water with very high initial turbidity. In summary coagulation efficiency of Moringa oleifera was found independent of storage temperature and container, however coagulation efficiency of Moringa oleifera decreased as storage duration increased. In addition, Moringa oleifera can be used as a potential coagulant especially for very high turbidity water. PMID- 16213138 TI - Selective and orally bioavailable phenylglycine tissue factor/factor VIIa inhibitors. AB - We describe the structure-based design and synthesis of highly potent, orally bioavailable tissue factor/factor VIIa inhibitors which interfere with the coagulation cascade by selective inhibition of the extrinsic pathway. PMID- 16213139 TI - Intermediate filament proteins participate in signal transduction. AB - How timely transport of chemical signals between the distal end of long axonal processes and the cell bodies of neurons occurs is an interesting and unresolved issue. Recently, Perlson et al. presented evidence that cleavage products of newly synthesized vimentin, an intermediate filament (IF) protein, interact with mitogen-activated protein (MAP) kinases at sites of axon injury. These IF fragments appear to be required for the transport of these kinases to the cell body along microtubule tracks. The truncated vimentin is instrumental in signal propagation as it provides a scaffold that brings together activated MAP kinases (such as Erk 1 and Erk2), as well as importin beta and cytoplasmic dynein. The authors propose that this all-in-one transport complex has the extraordinary ability to travel towards the cell body and enter the nucleus where the kinases activate and influence gene expression so that a neuron can generate a timely response to injury. PMID- 16213141 TI - Versatile synthesis and biological evaluation of 1,3-diamino-substituted 1alpha,25-dihydroxyvitamin D3 analogues. AB - A concise route to 1alpha,3beta-diamino-25-hydroxy-3-deoxyvitamin D(3) (5) and 1beta,3alpha-diamino-25-hydroxy-3-deoxyvitamin D(3) (6) has been developed starting from (R)- or (S)-carvone for the construction of the modified A-ring fragments. The conversion of the hydroxyl group to amine function with complete inversion of the configuration was efficiently accomplished by Mitsunobu reaction using phthalimide as nucleophile or activation of the hydroxyl group as mesylate followed by reaction with NaN(3). Diamino 5 and 6 as well as monoamino 3, 4, 30, and 31 vitamin D(3) derivatives have shown poor binding to VDR compared with 1alpha,25-dihydroxyvitamin D(3). The most active compound in the inhibition of MCF-7 cell proliferation and HL 60 cell differentiation was 1alpha-amino analogue 3. Also, very low in vivo calcemic effects of derivatives 3 and 4 were found. PMID- 16213140 TI - Effect of age on anticipatory postural adjustments in unilateral arm movement. AB - Aging is frequently accompanied by a deterioration in postural control. Accordingly, the elderly adopt postural strategies in order to maintain balance. The purpose of this study was to compare anticipatory postural adjustments in (healthy) 10 young and 10 elderly subjects using electromyography (EMG) and biomechanical parameters. While standing on a force platform, subjects performed voluntary, arm-raising movements under five conditions: self-paced at three different velocities, self-paced with load and an externally triggered, both at maximal velocity. The force platform provided information on vertical torque (T(z)) and center of pressure anteroposterior displacements (COP). EMG activity was recorded from the biceps femoris, quadriceps, tibialis anterior and soleus muscles. Voluntary movements were associated with an early COP backward shift and an anticipatory T(z). At low velocity, elderly subjects did not show any impairment in stability. At maximal velocity, T(z) was delayed in all conditions in the elderly group, whereas COP latency was reduced only in the self-paced condition without load. Despite this decrease in anticipation, the movement was performed at the same velocity as in younger subjects. The elderly adopted various muscle strategies in order to perform the same movement with less stability. In the self-paced condition, elderly subjects used a hip strategy, whereas young subjects used an ankle strategy. In the triggered condition, the strategy corresponded to increased activation of certain thigh muscles, rather than a sequence modification. Hence, local muscle strategies were used to counteract the overall delay in postural preparation revealed by biomechanical parameters. PMID- 16213142 TI - Synthesis of C-6 substituted pyrazolo[1,5-a]pyridines with potent activity against herpesviruses. AB - A novel series of potent C-6 substituted pyrazolo[1,5-a]pyridine inhibitors of herpes simplex viruses has been identified. A synthetic methodology was developed involving functionalization of a C-6 trifluoromethyl pyrazolo[1,5-a]pyridine to allow facile access to a diverse set of analogues from common late stage intermediates. The expansion of the SAR of this series at the 6 position allows for modifications to developability parameters such as clogP, while maintaining potency comparable to acyclovir. PMID- 16213143 TI - Exploration of acyl sulfonamides as carboxylic acid replacements in protease inhibitors of the hepatitis C virus full-length NS3. AB - The hepatitis C virus (HCV) NS3 protease has emerged as a promising anti-HCV drug target. Herein, we present an investigation of NS3 inhibitors comprising the acyl sulfonamide functionality. A series of tetra- and tripeptide based acyl sulfonamide inhibitors and their structure-activity relationships from both enzymatic and cell-based in vitro assays are presented. In summary, the acidity of the acyl sulfonamide functionality, the character of the P1 side chain, and the acyl sulfonamide substituent were found to be important for the inhibitory potencies. PMID- 16213144 TI - Development and optimization of a useful assay for determining Hsp90's inherent ATPase activity. AB - The Hsp90 molecular chaperone is responsible for the conformational maturation of nascent polypeptides and the rematuration of denatured proteins. Inhibition of Hsp90 represents a promising approach towards the treatment of cancer because numerous signaling cascades can be simultaneously targeted by disruption of the Hsp90-mediated process. Hsp90's ATPase activity is essential to the Hsp90 mediated protein folding process, consequently, a coupled assay was developed and optimized for determination of Hsp90's inherent ATPase activity. Using maltose phosphorylase, glucose oxidase, and horseradish peroxidase as components of this assay, a highly reproducible assay with a Z-factor of 0.87 has been produced. PMID- 16213145 TI - Design, synthesis, and evaluation of novel galloyl pyrrolidine derivatives as potential anti-tumor agents. AB - A series of novel galloyl pyrrolidine derivatives were synthesized as potential anti-tumor agents. Their inhibiting activities on gelatinase (MMP-2 and -9) were tested with succinylated gelatin as the substrate. Structure-activity analyses demonstrate that introduction of longer and more flexible side chains at the C(4) position of the pyrrolidine ring brings higher activity against gelatinase. Free phenol hydroxyl group is more favorable than the methylated one, which confirms the important role of the phenol hydroxyl group when inhibitors interact with gelatinase. In particular, (2S,4S)-4-(3-(3,4-dimethoxyphenyl)acrylamido)-N hydroxy-1-(3,4,5- trimethoxybenzoyl)pyrrolidine-2-carboxamide (18) stood out as the most attractive compound (IC(50) = 0.9 nM). The anti-metastasis model of mice bearing H(22) tumor cells was used to evaluate their anti-tumor activities in vivo. The assay in vivo revealed that most of these inhibitors displayed favorable inhibitory activities (inhibitory rate >35%) and no significant toxic effects were observed. The inhibition for 62.37% of 19 indicates the strategy used to design MMP inhibitors (MMPIs) of galloyl pyrrolidine derivatives as potential anti-tumor agents is promising. PMID- 16213146 TI - Crystallographic studies on two bioisosteric analogues, N-acetyl-beta-D glucopyranosylamine and N-trifluoroacetyl-beta-D-glucopyranosylamine, potent inhibitors of muscle glycogen phosphorylase. AB - Structure-based inhibitor design has led to the discovery of a number of potent inhibitors of glycogen phosphorylase b (GPb), N-acyl derivatives of beta-D glucopyranosylamine, that bind at the catalytic site of the enzyme. The first good inhibitor in this class of compounds, N-acetyl-beta-D-glucopyranosylamine (NAG) (K(i) = 32 microM), has been previously characterized by biochemical, biological and crystallographic experiments at 2.3 angstroms resolution. Bioisosteric replacement of the acetyl group by trifluoroacetyl group resulted in an inhibitor, N-trifluoroacetyl-beta-D-glucopyranosylamine (NFAG), with a K(i) = 75 microM. To elucidate the structural basis of its reduced potency, we determined the ligand structure in complex with GPb at 1.8 angstroms resolution. To compare the binding mode of N-trifluoroacetyl derivative with that of the lead molecule, we also determined the structure of GPb-NAG complex at a higher resolution (1.9 angstroms). NFAG can be accommodated in the catalytic site of T state GPb at approximately the same position as that of NAG and stabilize the T state conformation of the 280 s loop by making several favourable contacts to Asn284 of this loop. The difference observed in the K(i) values of the two analogues can be interpreted in terms of subtle conformational changes of protein residues and shifts of water molecules in the vicinity of the catalytic site, variations in van der Waals interaction, and desolvation effects. PMID- 16213147 TI - Studies on the chemistry of thienoanellated O,N- and S,N-containing heterocycles. Part 30: synthesis and pharmacological properties of thieno[2,3-b][1,4]thiazines with potential vasopressin receptor antagonistic activity. AB - A series of new nonpeptide vasopressin antagonists with a 6-ethyl-thieno[2,3 b][1,4]thiazine or 6-benzyl-thieno[2,3-b][1,4]thiazine skeleton and structural modifications of the aryl side chain were synthesized in this study. The effects on guinea pig heart and smooth muscle preparations were investigated. In the presence of AVP the compounds showed an antagonistic effect. The compounds did not change spontaneous rate in right atria and exerted a slight but not significant negative inotropic effect in papillary muscles. The relaxing effect on vascular smooth muscle and terminal ileum was far more pronounced. Generally the relaxing effect on terminal ilea was more potent maybe due to difference in V1a receptor density. Our results demonstrate that compounds with an ethyl group in position six on the thienothiazine ring (14, 16, 18 and 22) exerted the most potent relaxing activity in terminal ilea, whereas compounds with a phenyl ring in position six reduced this effect. PMID- 16213149 TI - The role of constitutively active Stat6 in leukemia and lymphoma. AB - Signal transducers and activators of transcription (STAT) are a family of transcription factors that regulate a broad range of cellular processes, such as proliferation, differentiation, and survival, in a large variety of cell types. Because of their regulation of diverse cellular functions, their aberrant activation is frequently associated with disease development, particularly oncogenic diseases. Much evidence exists to suggest that STAT proteins play a significant role in cellular transformation. However, which STAT proteins and to what extent they cause transformation and subsequent disease progression are topics currently being investigated. In this review, we will report on the findings concerning the involvement of Stat6 in the development of lymphoma and leukemia. Mounting evidence, in both patients and mouse models, supports a model where Stat6 is not a mere bystander, but rather, plays an active role in promoting a transformed phenotype. PMID- 16213148 TI - Myelin-associated mRNA and protein expression deficits in the anterior cingulate cortex and hippocampus in elderly schizophrenia patients. AB - Microarray and other studies have reported oligodendrocyte and myelin-related (OMR) deficits in schizophrenia. Here, we employed a quantitative approach to determine the magnitude of OMR gene expression deficits and their brain-region specificity. In addition, we examined how expression levels among the studied OMR genes are interrelated. mRNA of MAG, CNP, SOX10, CLDN11, and PMP22, but not MBP and MOBP, was reduced in the hippocampus and anterior cingulate cortex but not in the putamen of patients with schizophrenia. Expression of the only protein examined (CNP) was decreased in the hippocampus but not in the putamen. Correlation and factor analyses revealed that mRNA levels for genes that did exhibit differential expression in schizophrenia (MAG, CNP, SOX10, CLDN11, and PMP2), as opposed to those that did not (MOBP and MBP), loaded on separate factors. Thus, OMR gene and protein expression deficits in schizophrenia are brain-region specific, and the affected components may share regulatory elements. PMID- 16213150 TI - Monitoring of minimal residual disease in acute myeloid leukemia. AB - Monitoring minimal residual disease (MRD) becomes increasingly important in the risk-adapted management of patients with acute myeloid leukemia (AML). The two most sensitive and quantitative methods for MRD detection are multiparameter flow cytometry (MFC) and real-time polymerase chain reaction (QRT-PCR). Fusion gene specific PCR in AML is based on the RNA level, and thus in contrast to MFC expression levels rather than cell counts are assessed. For both methods independent prognostic values have been shown. The strong power of MFC has been shown mainly in the assessment of early clearance of the malignant clone. MRD levels in AML with fusion genes have the strongest prognostic power after the end of consolidation therapy. In addition, with QRT-PCR highly predictive initial expression levels can be assessed. With both methods early detection of relapse is possible. So far, validated PCR-based MRD was done with fusion genes that are detectable in only 20-25% of all AML MFC is superior since it is applicable for most AML. However, QRT-PCR is still more sensitive in most cases. Thus, it is desirable to establish new molecular markers for PCR-based studies. Large clinical trials will determine the role and place of immunologic and PCR-based monitoring in the prognostic stratification of patients with AML. PMID- 16213151 TI - Novel targeted therapies to overcome imatinib mesylate resistance in chronic myeloid leukemia (CML). AB - Imatinib mesylate (Gleevec) was developed as the first molecularly targeted therapy that specifically inhibits the BCR-ABL tyrosine kinase activity in patients with Philadelphia chromosome positive (Ph+) chronic myeloid leukemia (CML). Due to its excellent hematologic and cytogenetic responses, particularly in patients with chronic phase CML, imatinib has moved towards first-line treatment for newly diagnosed CML. Nevertheless, resistance to the drug has been frequently reported and is attributed to the fact that transformation of hematopoietic stem cells by BCR-ABL is associated with genomic instability. Point mutations within the ABL tyrosine kinase of the BCR-ABL oncoprotein are the major cause of resistance, though overexpression of the BCR-ABL protein and novel acquired cytogenetic aberrations have also been reported. A variety of strategies derived from structural studies of the ABL-imatinib complex have been developed, resulting in the design of novel ABL inhibitors, including AMN107, BMS-354825, ON012380 and others. The major goal of these efforts is to create new drugs that are more potent than imatinib and/or more effective against imatinib-resistant BCR-ABL clones. Some of these drugs have already been successfully tested in preclinical studies where they show promising results. Additional approaches are geared towards targeting the expression or stability of the BCR-ABL kinase itself or targeting signaling pathways that are chronically activated and required for transformation. In this review, we will discuss the underlying mechanisms of resistance to imatinib and novel targeted approaches to overcome imatinib resistance in CML. PMID- 16213152 TI - Modern diagnostics in acute leukemias. AB - Acute leukemias are a heterogeneous group of diseases. The different subtypes are characterized by certain clinical features and specific laboratory findings. Large clinical trials have confirmed the important impact of the underlying biology of each subtype for clinical outcome. Improvements in patient's treatment resulting in better survival rates are closely linked to the biological understanding of the disease subtypes, which is assessed by specific diagnostic approaches. Thus, several diagnostic techniques are mandatory at diagnosis for classification and for individual therapeutic decisions. Furthermore they are also needed for follow up studies focusing especially on minimal residual disease (MRD) to guide further treatment decisions based on the response of the disease to given treatment protocols. Only by using a comprehensive diagnostic panel including cytomorphology, cytochemistry, multiparameter flow cytometry (MFC), cytogenetics, fluorescence in situ hybridization (FISH) and molecular genetic methods the correct diagnosis in acute leukemias can be established today. The results serve as a mandatory prerequisite for individual treatment strategies and for the evaluation of treatment response using especially newly defined and highly specific MRD markers. PMID- 16213153 TI - Impact of comorbidity on treatment and prognosis of prostate cancer patients: a population-based study. AB - This study investigates the independent influence of serious comorbidity and age on treatment and survival of prostate cancer patients diagnosed between 1995 and 2002 in the southern part of the Netherlands. Eight percent of patients<60 years had two or more concomitant diseases versus 27% of those aged 80 years or older. The number of patients undergoing radical prostatectomy or curative radiotherapy decreased significantly with increasing age. The proportion of patients aged 60 69 years undergoing prostatectomy decreased significantly from 32% of patients without comorbidity to 17% of patients with two or more comorbid conditions and from 8% to 3%, respectively, of those aged 70-79 years. The risk of dying was significantly higher for patients with two or more comorbid conditions compared to patients without comorbidity. Serious comorbidity led to less aggressive treatment and negatively affected the prognosis of prostate cancer patients aged 60-79 years. PMID- 16213154 TI - Role of tumor necrosis factor receptors in an animal model of acute colitis. AB - TNF-alpha is known to play an important role in inflammatory bowel disease (IBD); however, the pathophysiological role of its receptors is still under study. Acute colitis was induced in rats by intracolonic administration of trinitrobenzene sulfonic acid (TNBS). Control rats received the ethanol vehicle. Rats were sacrificed 72 h later and samples of tissue and fluids were collected. There was a significant increase in the protein levels of sTNF-alpha, sTNFRI, and sTNFRII in the peritoneal fluid (PF) of experimental rats. TNF-alpha, TNFRI, and TNFRII mRNA expression was increased significantly in the colon of experimental animals compared to controls. TRAF3 and TRAF5 expression was also significantly higher, as was that of the adhesion molecules ICAM-1 and E-selectin. The increased expression of TNF-alpha, TNFRs, and the associated signaling factors in the colon of this rat model of IBD provides further evidence for their involvement in the promotion of inflammation and tissue damage. In addition, increased levels of sTNFRs in the PF of experimental rats--particularly sTNFRII--may be involved in the development of colitis by serving as a reservoir of TNF-alpha, and thus provide a novel therapeutic target for IBD. PMID- 16213155 TI - Contribution of the familial and genetic factors on monocyte chemoattractant protein-1 variation in healthy human pedigrees. AB - Monocyte chemoattractant protein-1 (MCP-1) is a chemokine whose circulating levels have been detected in the lesions of several diseases such as pulmonary fibrosis, rheumatoid arthritis and atherosclerosis. However, the factors involved in the regulation of its production remain largely unknown. The main aim of the present paper was to ascertain the contribution of the familial/genetic factors on the production of MCP-1 in apparently healthy individuals. We also tested the possible relationships between the plasma levels of MCP-1 and other cytokines involved in bone metabolism (receptor activator NF-kB ligand (RANKL), osteoprotegerin (OPG), interleukin-6, macrophage-colony stimulating factor, tumor necrosis factor-alpha). Using ELISA assays the cytokine levels were measured in 570 apparently healthy individuals belonging to ethnically homogeneous Caucasian families. We found that MCP-1 levels were significantly (P<0.01) correlated with RANKL (in both sexes) and with OPG only in women. The study showed that adjusted for potential covariates, 72% of the MCP-1 variance, was attributable to familial effects. About 49% was due to potential genetic factors and the rest was explained by common environmental sources shared by spouses within each family. In conclusion, our data provide reliable evidence for the substantial role of genetic factors in the determination of the phenotypic variability of MCP-1 plasma levels. The association between the osteoclastogenic cytokines and MCP-1 levels in healthy pedigrees is of special interest and might shed light on MCP-1 involvement in bone remodeling. PMID- 16213157 TI - Identification and characterization of a constitutively T-loop phosphorylated and active recombinant S6K1: expression, purification, and enzymatic studies in a high capacity non-radioactive TR-FRET Lance assay. AB - The p70 S6 ribosomal protein kinase 1 (S6K) is a substrate and effector of the mammalian target of rapamycin (mTOR). The mTOR/S6K pathway is implicated in cancer and metabolic disorders. To study the molecular regulation of S6K and identify specific inhibitors, availability of active recombinant S6K and robust enzyme assays are critically needed. To date, however, expression of active recombinant S6K has not been feasible as S6K activation requires a cascade of phosphorylation events. We have compared several engineered S6K enzymes. Expression of the Flag-S6KDeltaCT(T389E) in HEK293 cells resulted in a highly active S6K that was constitutively phosphorylated on T229 in the activation-loop (T-loop). The active enzyme was readily purified in large scale by anti-Flag affinity chromatography achieving a high purity. We developed a high capacity homogeneous time-resolved fluorescence resonance energy transfer. Lance assay for measurement of substrate phosphorylation and analysis of kinetic parameters. The Michaelis constant (Km) values of S6K for ATP and the Biotin-S6 substrate peptide were determined to be 21.4+/-0.29 and 0.9+/-0.48 microM, respectively. The Lance assay was further validated with a diverse panel of literature inhibitors, in which the PKC inhibitors staurosporine, Ro-318220, and the PKA inhibitor Balanol potently inhibited S6K. Dose-response and inhibition mechanism by these inhibitors were also studied. Our data provide a new simplified strategy to achieve rapid production of active S6K and demonstrate utility of the Lance assay for S6K enzyme screen in searching for specific inhibitors. PMID- 16213156 TI - Electrochemical molecular analysis without nucleic acid amplification. AB - Electrochemical biosensors have revolutionized glucose monitoring but have not yet fulfilled their promise of a low cost, direct detection replacement for genetic amplification tests such as PCR [K. Kerman, M. Kobayashi, E. Tamiya, Recent trends in electrochemical DNA biosensor technology, Meas. Sci. Technol. 15 (2004) R1-R11; A. Chaubey, B.D. Malhotra, Mediated biosensors. Biosens. Bioelectron. 17 (6-7) (2002) 441-456]. It has been anticipated that the integration of nanoscale chemical structures such as self-assembled monolayers with electrochemical biosensors would increase sensitivity by decreasing inherent system noise. We have designed a novel biosensing approach incorporating such integration and achieved rapid, ultra-low concentration sensitivities without target amplification. Raw samples are mixed with lysis buffer to allow hybridization of nucleic acid targets with anchor and signal probes before immobilizing a signaling enzyme proximate to the biosensor surface. A bias potential is subsequently applied and the secondary byproduct of a cyclic peroxidase reaction measured. Further studies have demonstrated the application of our approach in protein, clinical chemistry, and ionic assays. PMID- 16213158 TI - Partial characterization of mitogen-activated protein kinases (MAPK) from haemocytes of the common periwinkle, Littorina littorea (Gastropoda: Prosobranchia). PMID- 16213159 TI - Neuromuscular alterations during walking in persons with moderate knee osteoarthritis. AB - This paper compared the neuromuscular responses during walking between those with early-stage knee osteoarthritis (OA) to asymptomatic controls. The rationale for studying those with mild to moderate knee OA was to determine the alterations in response to dynamic loading that might be expected before severe pain, joint space narrowing and joint surface changes occur. We used pattern recognition techniques to explore both amplitude and shape changes of the surface electromyograms recorded from seven muscles crossing the knee joint of 40 subjects with knee OA and 38 asymptomatic controls during a walking task. The principal patterns for each muscle grouping explained over 83% of the variance in the waveforms. This result supported the notion that the main neuromuscular patterns were similar between asymptomatic controls and those with OA, reflecting the specific roles of the major muscles during walking. ANOVA revealed significant (p<0.05) differences in the principal pattern scores reflecting both amplitude and shape alterations in the OA group and among muscles. These differences captured subtle changes in the neuromuscular responses of the subjects with OA throughout different phases of the gait cycle and most likely reflected changes in the mechanical environment (joint loading, instability) and pain. The subjects with OA attempted to increase activity of the lateral sites and reduce activity in the medial sites, having minimal but prolonged activity during late stance. Therefore, alterations in neuromuscular responses were found even in this high functioning group with moderate knee OA. PMID- 16213160 TI - Distinguishing the evoked response from phase reset: a comment to Makinen et al. AB - Makinen et al. [Makinen, V., Tiitinen, H., May, P., 2005. Auditory event-related responses are generated independently of ongoing brain activity. Neuroimage 24, 961-968] suggest the use of amplitude variance to distinguish the evoked response from phase reset. Because their data do not exhibit a drop in amplitude variance, they conclude that ERPs are generated by 'processes separate from and additive to ongoing brain activity.' We argue that this conclusion is premature because of unrealistic assumptions about the processes underlying an event-related modulation of oscillations. A realistic phase reset model has to consider at least two parameters, degree of phase reset (or 'phase concentration') and amplitude change (event-related increase or decrease in amplitude). With simulated data, we show that a variable increase in amplitude size increases amplitude variance and masks the influence of a phase reset. On the other hand, an event-related decrease in amplitude without a phase reset leads to a sharp drop in variance. Furthermore, simulation of a frequency-specific phase reset shows a drop in variance that may be too small to be detected empirically. Thus, we conclude that amplitude variance is not capable of distinguishing the evoked response from phase reset. PMID- 16213161 TI - Network analysis of mild cognitive impairment. AB - We present a network analysis of a cross-sectional study of mild cognitive impairment (MCI). Network analysis, as opposed to univariate analysis, accounts for interactions among brain structures in explaining a clinical outcome. In this context, we analyze structural magnetic resonance (MR) data based on a Bayesian network representation of variables in the problem domain. The Bayesian network resulting from this analysis reveals complex, nonlinear multivariate associations among morphological changes in the left hippocampus and in the right thalamus and the presence of mild cognitive impairment. This Bayesian network could be used to predict the presence of mild cognitive impairment from structural MR scans. PMID- 16213162 TI - Abnormal brain activation during movement observation in patients with conversion paralysis. AB - Dissociative paralysis in conversion disorders has variably been attributed to a lack of movement initiation or an inhibition of movement. While psychodynamic theory suggests altered movement conceptualization, brain activation associated with observation and replication of movements has so far not been assessed neurobiologically. Here, we measured brain activation by functional magnetic resonance imaging during observation and subsequent imitative execution of movements in four patients with dissociative hand paralysis. Compared to healthy controls conversion disorder patients showed decreased activation of cortical hand areas during movement observation. This effect was specific to the side of their dissociative paralysis. No brain activation compatible with movement inhibition was observed. These findings indicate that in dissociative paralysis, there is not only derangement of movement initiation but already of movement conceptualization. This raises the possibility that strategies targeted at reestablishing appropriate movement conceptualization may contribute to the therapy of dissociative paralysis. PMID- 16213165 TI - Molecular insights on the taxonomic position of the paternal ancestor of the Squalius alburnoides hybridogenetic complex. PMID- 16213164 TI - Characterization of event-related designs using BOLD and IRON fMRI. AB - Despite many desirable characteristics, event-related (ER) stimulus designs for BOLD and IRON suffer from low detection power relative to block designs because the hemodynamic impulse response function (IRF) acts as a low-pass filter on neural activation to attenuate the size of differential responses to alternate stimuli. While the use of exogenous contrast agent (IRON technique) provides an alternative fMRI method in animal models to improve sensitivity and spatial localization, the inherently slower hemodynamic IRF causes IRON detection efficiency to decrease faster than BOLD efficiency as the interstimulus interval (ISI) is shortened. Using simulations based upon assumptions of stimulus-response linearity and experimental data obtained in awake, non-human primates, this study compared detection efficiencies for fixed, random and semi-random ISI distributions for BOLD and IRON techniques. A larger relative gain in detection efficiency at short ISI was obtained by randomized designs using IRON contrast relative to BOLD contrast due to the slower IRF of the IRON method. To quantify tradeoffs between detection efficiency and the predictability of stimulus presentation, the Shannon entropy was introduced as an objective measure of predictability. Small amounts of entropy can be traded for large gains in efficiency, particularly for the IRON method. PMID- 16213163 TI - Statistical diffusion tensor histology reveals regional dysmyelination effects in the shiverer mouse mutant. AB - Shiverer is an important model of central nervous system dysmyelination characterized by a deletion in the gene encoding myelin basic protein with relevance to human dysmyelinating and demyelinating diseases. Perfusion fixed brains from shiverer mutant (C3Fe.SWV Mbp(shi)/Mbp(shi)n = 6) and background control (C3HeB.FeJ, n = 6) mice were compared using contrast enhanced volumetric diffusion tensor magnetic resonance microscopy with a nominal isotropic spatial resolution of 80 mum. Images were accurately coregistered using non-linear warping allowing voxel-wise statistical parametric mapping of tensor invariant differences between control and shiverer groups. Highly significant differences in the tensor trace and both the axial and radial diffusivity were observed within the major white matter tracts and in the thalamus, midbrain, brainstem and cerebellar white matter, consistent with a high density of myelinated axons within these regions. The fractional anisotropy was found to be much less sensitive than the trace and eigenvalues to dysmyelination and associated microanatomic changes. PMID- 16213166 TI - On the Sigmodontinae radiation (Rodentia, Cricetidae): an appraisal of the phylogenetic position of Rhagomys. PMID- 16213168 TI - Systematics of the cyclostome subfamilies of braconid parasitic wasps (Hymenoptera: Ichneumonoidea): a simultaneous molecular and morphological Bayesian approach. AB - Phylogenetic relationships among 95 genera collectively representing 17 of the 18 currently recognized cyclostome braconid wasp subfamilies were investigated based on DNA sequence fragments of the mitochondrial COI and the nuclear 28S rDNA genes, in addition to morphological data. The treatment of sequence length variation of the 28S partition was explored by either excluding ambiguously aligned regions and indel information (28SN) or recoding them (28SA) using the 'fragment-level' alignment method with a modified coding approach. Bayesian MCMC analyses were performed for the separate and combined data sets and a maximum parsimony analysis was also carried out for the simultaneous molecular and morphological data sets. There was a significant incongruence between the two genes and between 28S and morphology, but not for morphology and COI. Different analyses with the 28SA data matrix resulted in topologies that were generally similar to the ones from the 28SN matrix; however, the former topologies recovered a higher number of significantly supported clades and had a higher mean Bayesian posterior probability, thus supporting the inclusion of information from ambiguously aligned regions and indel events in phylogenetic analyses where possible. Based on the significantly supported clades obtained from the simultaneous molecular and morphological analyses, we propose that a total of 17 subfamilies should be recognized within the cyclostome group. The subfamilial placements of several problematic cyclostome genera were also established. PMID- 16213167 TI - Comparison of evolutionary rates in the mitochondrial DNA cytochrome b gene and control region and their implications for phylogeny of the Cobitoidea (Teleostei: Cypriniformes). AB - It is widely accepted that mitochondrial DNA (mtDNA) control region evolves faster than protein encoding genes with few exceptions. In the present study, we sequenced the mitochondrial cytochrome b gene (cyt b) and control region (CR) and compared their rates in 93 specimens representing 67 species of loaches and some related taxa in the Cobitoidea (Order Cypriniformes). The results showed that sequence divergences of the CR were broadly higher than those of the cyt b (about 1.83 times). However, in considering only closely related species, CR sequence evolution was slower than that of cyt b gene (ratio of CR/cyt b is 0.78), a pattern that is found to be very common in Cypriniformes. Combined data of the cyt b and CR were used to estimate the phylogenetic relationship of the Cobitoidea by maximum parsimony, neighbor-joining, and Bayesian methods. With Cyprinus carpio and Danio rerio as outgroups, three analyses identified the same four lineages representing four subfamilies of loaches, with Botiinae on the basal-most clade. The phylogenetic relationship of the Cobitoidea was ((Catostomidae+Gyrinocheilidae)+(Botiinae+(Balitorinae+(Cobitinae+Nemacheilinae)) ), which indicated that Sawada's Cobitidae (including Cobitinae and Botiinae) was not monophyletic. Our molecular phylogenetic analyses are in very close agreement with the phylogenetic results based on the morphological data proposed by Nalbant and Bianco, wherein these four subfamilies were elevated to the family level as Botiidae, Balitoridae, Cobitidae, and Nemacheilidae. PMID- 16213169 TI - Determination of the rate of muscarinic M1 receptor plasma membrane delivery using a regulated secretion/aggregation system. AB - INTRODUCTION: In this study, we used the regulated secretion/aggregation technology (RPD) to determine the rate of human muscarinic M1 (hM1) receptor plasma membrane delivery. METHODS: hM1 receptors were expressed in CHO cells as C terminal fusion proteins to a conditional aggregation domain (CAD) consisting of four tandem mutant FKBP12 domains (F(m)). RESULTS: The CAD prevented the plasma membrane expression of hM1 receptors by causing the formation and intracellular retention of CAD-fused receptor aggregates as determined using intact cell [3H]NMS binding assays and epi-fluorescence microscopy, respectively. Aggregates of CAD-fused hM1 receptor could be disrupted in a concentration-dependent manner by the F(m)-selective ligand AP21998, resulting in an increased hM1 receptor plasma membrane expression. A furin cleavage site positioned between the CAD and the hM1 receptor sequence was cleaved by furin once aggregates of fusion protein were disrupted by AP21998, thus ensuring their irreversible dissolution. The plasma membrane delivery of hM1 receptors begins within 30 min of AP21998 exposure and the rate of delivery was constant for up to eight hours. In the continued presence of AP21998, hM1 receptor plasma membrane expression continued to increase for up to 18 h, then began to decrease toward basal levels as incubation continued out to 72 h. Using mathematical models, we determined the rate constants for the plasma membrane delivery of hM1 receptors from these data. Also, hM1 receptors elicited phosphoinositide hydrolysis to carbachol once expressed at the plasma membrane and the pharmacology of the response varied depending upon the concentration of AP21998 used to cause plasma membrane expression. DISCUSSION: Overall, our data indicate that the RPD can be used to characterize the kinetics of receptor plasma membrane delivery and to characterize functional responses elicited to different numbers of plasma membrane expressed receptor. PMID- 16213170 TI - Using solid-state 31P{19F} REDOR NMR to measure distances between a trifluoromethyl group and a phosphodiester in nucleic acids. AB - REDOR is a solid-state NMR technique frequently applied to biological structure problems. Through incorporation of phosphorothioate groups in the nucleic acid backbone and mono-fluorinated nucleotides, 31P{19F} REDOR has been used to study the binding of DNA to drugs and RNA to proteins through the detection of internuclear distances as large as 13-14 A. In this work, 31P{19F} REDOR is further refined for use in nucleic acids by the combined use of selective placement of phosphorothioate groups and the introduction of nucleotides containing trifluoromethyl (-CF3) groups. To ascertain the REDOR-detectable distance limit between an unique phosphorous spin and a trifluoromethyl group and to assess interference from intermolecular couplings, a series of model compounds and DNA dodecamers were synthesized each containing a unique phosphorous label and trifluoromethyl group or a single 19F nucleus. The dipolar coupling constants of the various 31P and 19F or -CF3 containing compounds were compared using experimental and theoretical dephasing curves involving several models for intermolecular interactions. PMID- 16213171 TI - Symmetric spatial encoding in ultrafast 2D NMR spectroscopy. AB - Single-scan multidimensional spectroscopy utilizes spatial dimensions for encoding the indirect-domain internal spin interactions. Various strategies have been hitherto demonstrated for fulfilling the encoding needs underlying this methodology; in analogy with their time-domain counterparts all of them have in common the fact that they proceed monotonically-starting at one end of the sample and concluding at the other. The present manuscript discusses another possibility that arises for the case of amplitude-modulated ultrafast nD NMR, whereby the spatial encoding progresses from both ends of the sample simultaneously towards the center. Such symmetric encoding is compatible with continuous or discrete excitations as well as with homonuclear or heteronuclear correlations, and exhibits a number of advantages vis-a-vis the unidirectional encodings that have been used so far: it originates echoes that are free from large first-order phase distortions, and yields nD peaks possessing a purely-absorptive character. It has the added advantage that for a given indirect-domain spectral resolution it can complete its task in half the time required by a conventional monotonic spatial encoding, leading to potentially important gains in sensitivity. The main features underlying this new spatially symmetric encoding protocol are derived, and its advantages are demonstrated with a series of amplitude-modulated homo- and hetero-nuclear 2D ultrafast NMR examples. PMID- 16213172 TI - Rectifying system-specific errors in NMR relaxation measurements. AB - 15N spin relaxation parameters provide a powerful tool for probing the internal dynamics and thermodynamics of proteins. The biological insight provided by these experiments often involves interpretation of small changes in relaxation parameters. This, in turn, requires careful data analysis, especially in the identification and treatment of systematic error. While progress continues on reduction of experiment-specific errors associated with pulse sequences, system specific sources of error have received far less attention. The impact of these errors varies between facilities, spectrometers, and biological samples. We demonstrate that performing a series of control experiments along with relaxation measurements can help identify, quantify, and isolate sources of system-specific error, and, in some cases, correct for systematic changes. We further demonstrate that control experiments can be performed without significant loss of spectrometer time, and lead to more accurate relaxation parameter values. PMID- 16213173 TI - Heterozygous mutations of growth hormone receptor gene in children with idiopathic short stature. AB - OBJECTIVE: The term idiopathic short stature (ISS) describes children: (a) whose height is more than two standard deviations below the mean; (b) with normal or slow height velocity; (c) of normal birth weight; (d) showing an absence of specific endocrine abnormalities; and (e) having no evidence of chronic physical or psychological illness. It has been suggested that partial growth hormone (GH) insensitivity due to heterozygous mutations of the GH Receptor gene may account for some cases of ISS. DESIGN AND METHODS: GHR gene was investigated (SSCP assay and direct sequencing) in 37 ISS patients. Fifty controls were recruited from the same geographic area as the patients; age and gender were stratified to match controls to patients. RESULTS: We observed the previously described transition A>G (GGA>GGG) of position 3 of codon 168, determining the synonymous change G168G in 22 of 37 patients (12 homozygous and 10 heterozygous) and in 23 of 50 controls (16 homozygous and 7 heterozygous). The relative allele frequency was similar in patients and in controls. In one ISS patient we identified a novel transition T>C (TGT>TGC) of position 3 of codon 94 , determining the synonymous change C94C. In another patient we demonstrated a novel heterozygous transition T>C (GTC>GCC) of the position 2 of codon 144, determining the missense mutation V144A, These mutations were not found in 100 control chromosomes. CONCLUSIONS: Heterozygous mutations of the GHR gene are uncommon in Italian ISS patients, who are selected for adequate GH levels. However the observed incidence of 2 mutations out of 37 ISS patients (i.e., 5%) is not different from the one previously reported in the literature. PMID- 16213174 TI - GH secretion in acute exercise may result in post-exercise lipolysis. AB - Exercise is a potent stimulator of growth hormone (GH) secretion. We hypothesised that after a short bout of intense exercise GH may increase lipolysis during recovery. In 7 moderately trained young male subjects (21.8 +/- 0.5 years) and 7 moderately trained older male subjects (56.0 +/- 1.0 years) [(2)H(5)] glycerol was infused for 370min to measure glycerol production rate (R(a)), a measure of lipolysis. At 130 min subjects exercised on a cycle ergonometer for 20 min at 70% V(O2 max), followed by rest for 220 min. On a separate occasion the study was repeated in the young subjects with a 1h GH infusion (4microgkg(-1)h(-1)) at 130 min instead of exercise. In response to exercise, catecholamines (p < 0.02) and glycerol R(a) (p < 0.01) increased, peaking during exercise. GH concentration increased in response to exercise (p < 0.01), peaking after exercise (150-160 min) in both groups with no significant difference in peak response between groups. A post-exercise rise in glycerol R(a) was demonstrated in both groups peaking at 265-295 min in the older group (p < 0.002, peak vs. basal) and continuing to rise until 370 min in the young group (p < 0.01, peak vs. basal). The timing and magnitude of this was reproduced with the GH infusion. There was a significant correlation between the peak GH response to exercise and the post exercise rise in glycerol R(a) measured as area under the curve (r=0.57, p < 0.04). In conclusion, this study provides evidence that the GH response to acute exercise may increase lipolysis during recovery. PMID- 16213175 TI - Vaccination equally enables both genetically susceptible and resistant mice to control infection with group A streptococci. AB - There is substantial evidence that host genetic factors are important in determining susceptibility to infection with group A streptococci (GAS). Several studies have revealed that, similarly to humans, a genetic component may be important in determining susceptibility to GAS infection in mice. Thus, C3H/HeN mice are much more susceptible to streptococcal infection than BALB/c mice. We have determined here whether vaccination makes genetically susceptible mice as capable as genetically resistant mice to control GAS infection. Resistant BALB/c and susceptible C3H/HeN mice were immunized either systemically with heat-killed GAS or through the mucosal route with an M protein-based subunit vaccine, and challenged with live bacteria. Vaccination elicited in both mouse strains similar levels of bactericidal anti-GAS IgG antibodies and also antigen-specific mucosal IgA. Vaccination provided mice of both strains with an increased and equal capacity to express immunity against GAS as indicated by the reduced level of bacteria in the organs and the ability of vaccinated mice to survive infection. Protection in vaccinated mice was dependent on the presence of T cell-dependent bactericidal antibodies as shown by the ability of serum elicited in immunocompetent mice but not of serum elicited in T cell-deficient nu/nu mice to passively transfer anti-GAS immunity. In conclusion, the results presented here demonstrated that the presence of anti-GAS specific, T cell-dependent bactericidal antibodies elicited after vaccination overcomes the innate genetic susceptibility of C3H/HeN mice and makes both resistant and susceptible mice equally capable of controlling GAS infection. PMID- 16213176 TI - Immune stimulation and malaria infection impose reproductive costs in Anopheles gambiae via follicular apoptosis. AB - The employment of defense mechanisms is recognized as a costly life-history trait. In the malaria vector Anopheles gambiae, reproductive costs have been associated with both humoral and cellular innate immune responses and also with malaria infection. The resorption of developing oocytes associated with malaria infection is preceded by the programmed cell death, or apoptosis, of follicular cells. Here we demonstrate that apoptosis in ovarian follicular epithelial cells also occurs when mosquitoes are subjected to artificial immune-elicitors that induce a melanization response or humoral antimicrobial activity. Caspases are key cysteine proteases involved in apoptosis. Caspase-like activity was detected in epithelial cells in approximately 4.0% of the developing ovarian follicles of untreated, blood-fed, mosquitoes. Lipopolysaccharide injection resulted in a significant increase in anti-Micrococcus luteus humoral activity and a significant increase of 257.7% of follicles exhibiting apoptosis compared to results after saline injections. Melanization also triggered follicular apoptosis, which increased by 106.25% or 134.37% in Sephadex C-25 or G-25 bead inoculated mosquitoes, respectively, compared to that in sham-injected ones. Ovaries from Plasmodium yoelii nigeriensis-infected mosquitoes exhibited a significant increase in follicular apoptosis of 440.9% compared to non-infected ones. Thus, at the time point investigated, infection had a much greater effect than artificial immune-elicitors. Death of follicular epithelial cells has been shown to lead to follicle resorption and hence a decrease in egg production. We propose the trade-off between reproductive fitness and immune defense in A. gambiae operates via the induction of apoptosis in ovarian follicles and that different immune responses impose costs via the same pathway. PMID- 16213178 TI - Anti-tumor DNA vaccines based on the expression of human papillomavirus-16 E6/E7 oncoproteins genetically fused with the glycoprotein D from herpes simplex virus 1. AB - DNA vaccines encoding the human papillomavirus type-16 (HPV-16) E6 and E7 oncoproteins genetically fused to the human herpes simplex virus type 1 (HSV-1) gD protein were tested in mice for induction of T cell-mediated immunity and protection against tumor cell challenge. Hybrid genes, generated after insertion of E6 or E7-encoding sequences into internal sites of the gD-encoding gene, were transcribed in vitro and the chimeric proteins were expressed at the surface of in vitro-transfected mammalian cells. Female C57BL/6 mice immunized with 4 intramuscular doses (100 microg of DNA/dose) of the DNA vaccines encoding E7 efficiently generated E7-specific CD8(+) T cells. Vaccination of mice with the DNA vaccines encoding the E7, or both E6 and E7, conferred complete protection to challenges from TC-1 tumor cells and partial therapeutic effect (40%) in mice inoculated with TC-1 cells on the same day or 5 days prior to the first vaccine dose. PMID- 16213179 TI - Melanin in the dimorphic fungal pathogen Paracoccidioides brasiliensis: effects on phagocytosis, intracellular resistance and drug susceptibility. AB - The fungal pathogen Paracoccidioides brasiliensis produces a melanin-like pigment in the presence of l-DOPA in vitro. We investigated whether melanization affected yeast uptake by alveolar and peritoneal macrophages, the intracellular resistance of fungal cells and their susceptibility to antifungal drugs. The interactions of melanized and nonmelanized P. brasiliensis with murine primary macrophages and J774.16 and MH-S macrophage-like cell lines were investigated. Melanized yeast cells were poorly phagocytosed by the cells even in the presence of complement. Melanization caused significant interference with the binding of cell wall components to lectin receptors on macrophages. Melanized cells were also more resistant than nonmelanized cells to the antifungal activity of murine macrophages. No difference in the susceptibilities of melanized and nonmelanized P. brasiliensis to antifungal drugs was observed using the minimum inhibitory concentration (MIC) method. However killing assays showed that melanization significantly reduced fungal susceptibility to amphotericin B and also protected against ketoconazole, fluconazole, itraconazole and sulfamethoxazole. The present results indicate that fungal melanin protects P. brasiliensis from phagocytosis and increases its resistance to antifungal drugs. PMID- 16213180 TI - Co-infection with two different Campylobacter jejuni strains in a patient with the Guillain-Barre syndrome. AB - Campylobacter jejuni is the predominant cause of antecedent infection in Guillain Barre syndrome (GBS) or Miller Fisher syndrome (MFS). C. jejuni probably triggers GBS or MFS through molecular mimicry between bacterial sialylated lipo oligosaccharides (LOS) and gangliosides in peripheral nerve tissue. We investigated whether co-infections with multiple C. jejuni strains occur in GBS or MFS patients and we further characterized these strains. PFGE analysis of 83 C. jejuni isolates from single primary colonies from stool cultures of 13 patients with GBS or MFS revealed co-infection with two different strains in one patient (8%). We showed that only strain GB5.1 contained an LOS biosynthesis gene locus that is associated with neuropathy. The patient serum strongly reacted with the LOS of strain GB5.1 and not with the LOS of strain GB5.2. Mass spectrometry revealed that both strains expressed a non-sialylated outer core structure in their LOS. The patient serum contained anti-asialo-GM2 antibodies that cross reacted with the LOS of strain GB5.1. This study demonstrates that co-infection with multiple C. jejuni strains occurs in GBS patients. Consequently, not all C. jejuni strains isolated from the faeces of a GBS patient are involved in the pathogenesis of GBS per se. Furthermore, this is the first report in which cross reactivity of antibodies to asialo-GM2 and to the LOS of a C. jejuni strain from a GBS patient has been demonstrated. This finding suggests that molecular mimicry with non-sialylated structures may also be involved in the pathogenesis of GBS. PMID- 16213181 TI - Criteria for selection of HIV vaccine candidates--general principles. AB - The generation of a vaccine against HIV/AIDS is extremely challenging, as evidenced by more than 20 years of attempts. Here are highlighted the strategies adopted within the AIDS Vaccine Integrated project (AVIP) to speed up the clinical evaluation of novel vaccine candidates and to increase the chances to get an effective preventive and/or therapeutic vaccine. PMID- 16213182 TI - The Toxoplasma surface protein SAG1 triggers efficient in vitro secretion of chemokine ligand 2 (CCL2) from human fibroblasts. AB - Chemokines play an important role in the physiopathology of toxoplasmosis in murine models. Infection of different human cell types by Toxoplasma gondii induces the secretion of these immune mediators. The aim of our study was to identify parasite molecules that could be involved in the triggering of chemokine ligand 2 (CCL2) secretion during T. gondii host cell invasion: surface, micronemal, rhoptry and dense granule proteins. The secretion of CCL2 was studied 1) after infection of human fibroblasts with mutants of Toxoplasma RH strain deficient either for GRA5, GRA2-GRA6, ROP1 or SAG1; 2) after stimulation by micronemal proteins or by the immunodominant surface antigen 1 of T. gondii. CCL2 secretion was quantified by ELISA at 3 h and/or 24 h after infection or stimulation. Infection by Deltagra2-Deltagra6, Deltagra5 or Deltarop1 mutants did not modify the level of CCL2, as compared with the level measured after infection with the wild-type strain. Moreover, stimulation with micronemal proteins did not increase the secretion of this chemokine. By contrast, the level of CCL2 was increased 3 h post-stimulation by purified or recombinant SAG1. Specificity of this effect was confirmed by the decrease in CCL2 secretion when human fibroblasts were infected with the Deltasag1 mutant (48%) as compared with the wild-type strain (100%). In conclusion, this major Toxoplasma surface protein SAG1, specific to the tachyzoite stage, is directly or indirectly involved in the cellular mechanisms triggering CCL2 secretion after T. gondii infection. These results could explain the parasitic mechanisms leading to cell infiltrates detected only in the presence of tachyzoites, a phenomenon observed in toxoplasmic reactivation. PMID- 16213183 TI - Rational vaccine strategies against AIDS: background and rationale. AB - New vaccine candidates exploiting the rational combination of regulatory and structural HIV gene products are being developed within the program of the AIDS Vaccine Integrated Project (AVIP) and will be tested in comparative preclinical and clinical trials with the ultimate goal of selecting proper candidates for advanced clinical testing in developing countries. PMID- 16213184 TI - Protection against Helicobacter pylori infection in the Mongolian gerbil after prophylactic vaccination. AB - Vaccines against Helicobacter pylori could circumvent the problem of increasing antibiotic resistance. They would be particularly useful in developing countries, where re-infection rates are high following standard eradication regimes. The Mongolian gerbil is a good model for H. pylori infection, as the gastric pathology induced by infection is similar to that in humans. The H. pylori induced inflammatory response in gerbils is considerably greater than in murine models. The aim of this study was to determine if gerbils could be vaccinated against H. pylori. Mongolian gerbils were vaccinated orally with an H. pylori whole cell sonicate preparation and cholera toxin adjuvant. Vaccinated gerbils and controls were challenged with the autologous H. pylori strain 42GX. All infection, and cholera toxin, control gerbils were H. pylori positive 6 weeks post-challenge. By contrast, a significant degree of protection was demonstrated in vaccinated gerbils. Only two of 10 of gerbils were H. pylori positive (P<0.001). Protection was associated with increased serum H. pylori IgG antibodies. Protected gerbils had histologically normal gastric mucosa and, in contrast to mice, no post-immunisation gastritis was evident. In the control groups, the degree of inflammation was variable, with some of the animals having corpus gastritis and corpus mucous metaplasia. The levels of gastric IL-12p40 and IFNgamma transcripts were significantly decreased in vaccinated animals compared to infection and cholera toxin controls (P<0.01). Gastric IL-10 and TGFbeta transcripts were found only at relatively low levels. These results demonstrate that Mongolian gerbils can be successfully vaccinated against H. pylori and protected from H. pylori-induced pathology. PMID- 16213185 TI - Troponin C/calmodulin chimeras as erythrocyte plasma membrane Ca2+-ATPase activators. AB - Calmodulin (CaM) and troponin C (TnC) are EF-hand proteins that play fundamentally different roles in animal physiology. TnC has a very low affinity for the plasma membrane Ca2+-ATPase and is a poor substitute for CaM in increasing the enzyme's affinity for Ca2+ and the rate of ATP hydrolysis. We use a series of recombinant TnC (rTnC)/CaM chimeras to clarify the importance of the CaM carboxyl-terminal domain in the activation of the plasma membrane Ca2+ ATPase. The rTnC/CaM chimera, in which the carboxyl-terminal domain of TnC is replaced by that of CaM, has the same ability as CaM to bind and transmit the signal to Ca2+ sites on the enzyme. There is no further functional gain when the amino-terminal domain is modified to make the rTnC/CaM chimera more CaM-like. To identify which regions of the carboxyl-terminal domain of CaM are responsible for these effects, we constructed the chimeras rTnC/3CaM and rTnC/4CaM, where only one-half of the C-terminal domain of CaM (residues 85-112 or residues 113-148) replaces the corresponding region in rTnC. Neither rTnC/3CaM nor rTnC/4CaM can mimic CaM in its affinity for the enzyme. Nevertheless, with respect to the signal transduction process, rTnC/4CaM, but not rTnC/3CaM, shows the same behaviour as CaM. We conclude that the whole C-terminal domain is required for binding to the enzyme while Ca2+-binding site 4 of CaM bears all the requirements to increase Ca2+ binding at PMCA sites. Such mechanism of binding and activation is distinct from that proposed for most other CaM targets. Furthermore, we suggest that Ala128 and Met124 from CaM site 4 may play a crucial role in discriminating CaM from TnC. PMID- 16213186 TI - The "Cinderella effect" is no fairy tale. PMID- 16213187 TI - Cytogenetic evaluation of arsenic trioxide toxicity in Sprague-Dawley rats. AB - Acute exposure to arsenic trioxide has been reported to induce death and/or multiple organ damage with symptoms including nausea, vomiting, diarrhea, gastrointestinal hemorrhage, cerebral edema, tachycardia, dysrhythmias and hypovolemic shock. Its toxic effects are due to its ability to bind to sulfhydryl groups of proteins and to inhibit energy production. Although the chronic exposure to arsenic trioxide has been linked to various types of cancer, such as skin, liver, lung, bladder and kidney neoplasms, studies of its carcinogenic potential in animals have not been conclusive. In this study, we investigated the genotoxic potential of arsenic trioxide in bone-marrow cells obtained from Sprague-Dawley rats; using chromosomal aberrations (CA), mitotic index (MI) and micronuclei (MN) formation as the toxicological endpoints. Four groups of six male rats each, weighing approximately 60+/-2 g per rat, were injected intraperitoneally, once a day for 5 days with doses of 5, 10, 15 and 20 mg/kg body weight (BW) of arsenic trioxide dissolved in distilled water. A control group was also made of six animals injected with distilled water without chemical. All the animals were sacrificed at the end of the treatment period. Chromosome and micronuclei preparation was obtained from bone-marrow cells following standard protocols. Arsenic trioxide exposure significantly increased the number of structural chromosomal aberrations, the frequency of micronucleated cells and decreased the mitotic index in treated groups when compared with the control group. Our results demonstrate that arsenic trioxide has a clastogenic/genotoxic potential as measured by the bone-marrow CA and MN tests in Sprague-Dawley rats. PMID- 16213188 TI - Modeling the dynamics and control of Schistosoma japonicum transmission on Bohol island, the Philippines. AB - We have investigated a mathematical model for the transmission of Schistosoma japonicum in the infested region of northeastern Bohol island in the Philippines. The development of transmission models is important for planning control strategies. Since S. japonicum has a complicated mode of transmission, the rates of transmission among its hosts cannot be measured directly by field observation. Instead, they have been estimated through model analysis. The model takes into account the seasonal variations and includes a function of control measures. In 1981, a project to eliminate schistosomiasis started on Bohol island. The prevalence decreased dramatically and has kept low level less than 1%. The simulations based on the model predicted that there is little probability of resurgence of an epidemic in the northeastern endemic villages of Bohol island due to the fact that the project has attained a high coverage of selective mass treatment based on stool examination accompanied by a successful snail control operation. PMID- 16213189 TI - Primary healthcare information system--the cornerstone for the next generation healthcare sector in Republic of Croatia. AB - At no time in the history of medicine has the growth in knowledge and technologies been so profound [Crossing the Quality Chasm: A New Health System for the 21st Century, Institute of Medicine (IOM), 2001. ISBN 0-309-07280-8]. However, healthcare delivery systems today are not able to keep up with the pace. Studies have shown that it takes an average of about 17 years for new knowledge generated by randomized trials to be incorporated into practice [B. Andrew, S. Boren, Managing clinical knowledge for health care improvement, in: Yearbook of Medical Informatics, National Library of Medicine, Bethesda, MD, 2000, pp. 65 70]. It is safe to say that today healthcare systems "have the data, but not information". In order to provide highest quality patient care, Republic of Croatia has started the process of introducing enterprise information systems to support business processes in the healthcare domain. Two major requirements are in focus: to provide efficient healthcare related data management in support of decision-making processes; and to support continuous process of healthcare resources spending optimization. The first initiated project refers to Primary Healthcare Information System (PHCIS) that provides domain of primary care with state-of-the-art enterprise information system that connects General Practitioners, Pediatricians and Gynecologists offices with the Croatian Institute for Health Insurance and Public Health Institute. In the years to come, PHCIS will serve as the main integration platform for connecting all other stakeholders and levels of healthcare (e.g. hospitals, pharmacies, laboratories) into single enterprise healthcare network. This article gives an overview of PHCIS, explains challenges that were faced in designing and implementing the system, and elaborates PHCIS role as the cornerstone for the next generation healthcare provisioning in Republic of Croatia. PMID- 16213190 TI - Application of 31P MRS to the analysis of phospholipid changes in plasma of patients with acute leukemia. AB - The aim of the experiment was to evaluate the changes of phospholipid concentrations in patients (n=30) with acute leukemia compared with reference group of healthy volunteers (n=21). The analysis focused on the following phospholipids (PL) collected from plasma: phosphatidylcholine (PC), plasmalogen of phosphatidylcholine (CPLAS), lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylethanolamine (PE), and phosphatidylinositol (PI). Phospholipid extracts were obtained by Folch's method from 4 ml of plasma. 31P MR spectra were obtained on an AMX Bruker 300 MHz (7.05 T) spectrometer. Calculation of concentration based on integral intensity of the phospholipid relative to an internal concentration standard of MDPA. For healthy volunteers, the following values of phospholipid concentrations were obtained: (5.18+/-1.615) mmol/l for PC+CPLAS; (0.364+/-0.178) mmol/l for LPC; (1.211+/-0.411) mmol/l for SM; (0.343+/ 0.124) mmol/l for PI+PE. PLs of patients were assayed at least twice: at the time of diagnosis and, when appropriate, at the time of complete remission from the disease (CR). At the time of diagnosis, the mean concentrations of studied compounds were: (1.602+/-0.716) mmol/l for PC+CPLAS; (0.041+/-0.048) mmol/l for LPC; (0.398+/-0.198) mmol/l for SM; (0.045+/-0.071) mmol/l for PI+PE. After attainment of complete remission (CR), the respective values were as follows: (4.094+/-1.886) mmol/l for PC+CPLAS; (0.295+/-0.139) mmol/l for LPC; (1.123+/ 0.634) mmol/l for SM; (0.230+/-0.125) mmol/l for PI+PE. All concentrations found in patients at the time of diagnosis were significantly lower than in reference group and in those benefited from complete remission (CR). By contrast the differences in concentrations of phospholipids in plasma between patients with complete remission (CR) and healthy volunteers were no statistically significant. PMID- 16213191 TI - Mesenchymal stem cell therapy to rebuild cartilage. AB - Disorders affecting cartilage touch almost the whole population and are one of the leading causes of invalidity in adults. To repair cartilage, therapeutic approaches initially focused on the implantation of autologous chondrocytes, but this technique proved unsatisfactory because of the limited number of chondrocytes obtained at harvest. The discovery that several adult human tissues contain mesenchymal stem cells (MSCs) capable of differentiating into chondrocytes raised the possibility of injecting MSCs to repair cartilages. The important data published recently on the factors controlling chondrocyte commitment must be thoroughly considered to make further progress towards this therapeutic approach. The potential application of MSC therapy provides new hope for the development of innovative treatments for the repair of cartilage disorders. PMID- 16213192 TI - Adenovirus-mediated gene transfer and lipoprotein-mediated protein delivery of plasma PAF-AH ameliorates proteinuria in rat model of glomerulosclerosis. AB - Oxidative stress has been proposed to play a crucial role in glomerulosclerosis, although its in vivo demonstration has proved taxing given the difficulty of inducing gene expression in specific renal cells. In this study, we examined whether the liver-directed expression of plasma platelet-activating factor acetylhydrolase (PAF-AH) would affect the glomerular pathophysiology in Imai rats, an animal model for glomerulosclerosis. Adenovirus-mediated liver-directed gene delivery of human PAF-AH resulted in a significant increase in plasma PAF-AH activity, which was detected almost exclusively on HDL. Histological examination of rats overexpressing PAF-AH showed not only the deposition of PAF-AH in mesangial cells, but also a reduction in hydroxynonenal and matrix protein content in the glomeruli. In situ hybridization analysis was negative for human PAF-AH mRNA in the kidney, while injection of HDL abundant in PAF-AH resulted in the deposition of PAF-AH in mesangial cells. Urine protein levels did not increase in rats overexpressing PAF-AH, while those of control rats increased significantly with age. This study provides direct evidence of the in vivo role of an enzyme that degrades lipid peroxides during the progression of glomerulosclerosis. Adenovirus-mediated extrarenal gene expression and lipoprotein-mediated glomeruli-targeted protein delivery promise to be a novel therapeutic approach to glomerulosclerosis. PMID- 16213193 TI - Influence of sympathetic tone on heart rate during vagal stimulation and nitroprusside induced hypotension in ovine fetus. AB - OBJECTIVE: To characterize effects of sympathetic tone on fetal heart rate (FHR) reflex responses and FHR variability in late gestation. DESIGN/METHODS: Changes in FHR and autonomic tones were studied (i) after electrical vagal stimulation and (ii) during nitroprusside-induced hypotension, in seven late gestation ovine fetus in control condition (ctrl), after dobutamine (beta1-activation) and atenolol (beta1-blockade). Results are expressed as mean +/- SEM. RESULTS: (i) Minimal FHR after vagal stimulation was not influenced by atenolol or dobutamine but dobutamine accelerated FHR normalization. (ii) During nitroprusside induced hypotension atenolol inhibited the initial increases in FHR and FHR variability (measured by SD and LFnu) but not the bradycardia occurring below a mean arterial pressure of 38 +/- 2 mmHg. Dobutamine did not abolish the depressor reflex. During hypotension the positive chronotropic effect of sympathetic tone increased from 15 +/- 2 to 42 +/- 7 bpm then decreased at a rate of -7.6 +/- 1.5 bpm mmHg( 1), vagal negative chronotropic influence steadily increased at a rate of 1.9 +/- 0.4 bpm mmHg(-1). Changes in FHR variability were not correlated with vagal or sympathetic chronotropic effects. CONCLUSIONS: beta1-stimulation does not affect sinus-node response to vagal stimulation but improves the speed of FHR normalization. FHR response to hypotension depends on an initial increase in both sympathetic and parasympathetic chronotropic effects that is associated with a sympathetic dependent increase in FHR variability and is followed by a withdrawal of sympathetic tone. PMID- 16213194 TI - Structure-function studies of DNA polymerase lambda. AB - DNA polymerase lambda is a member of the X family of polymerases that is implicated in non-homologous end-joining of double-strand breaks in DNA and in base excision repair of DNA damage. To better understand the roles of DNA polymerase lambda in these repair pathways, here we review its structure and biochemical properties, with emphasis on its gap-filling polymerization activity, its dRP lyase activity and its unusual DNA synthetic (in)fidelity. PMID- 16213195 TI - Development of specific Rho-kinase inhibitors and their clinical application. AB - Hexahydro-1-(isoquinoline-5-sulfonyl)-1H-1,4-diazepine, HA-1077, is a known selective inhibitor of Rho-kinase. Although its IC(50) value against Rho-kinase is more than 10 times lower than those for kinases such as PKA, PKB, PKC, PKG, MLCK, CaMKII and others, the molecule still retains relative potent inhibition activities against these kinases. In order to produce highly specific Rho-kinase inhibitors, several HA-1077 analogs were synthesized and their kinase inhibition properties evaluated. (S)-Hexahydro-1-(4-ethenylisoquinoline-5-sulfonyl)-2-methyl 1H-1,4-diazepine was found to be a potent Rho-kinase inhibitor. The IC50 value against Rho-kinase was 6 nM, while those against other kinases remained at almost the same level as that of HA-1077. Furthermore, we designed HA-1077 analogs on the basis of the complex structure of PKA and HA-1077. Amongst these, (S) hexahydro-4-glycyl-2-methyl-1-(4-methylisoquinoline-5-sulfonyl)-1H-1,4-diazepine and other glycine derivatives were found to be highly specific Rho-kinase inhibitors. These Rho-kinase specific inhibitors were applied to rabbit ocular hypertensive models and were shown to reduce intraocular pressure. These results demonstrate that the new 5-isoquinolinesulfonylamides are not only potent ROCK selective compounds, but are also useful compounds for clinical applications. PMID- 16213196 TI - Crystal structure of alkyl hydroperoxide-reductase (AhpC) from Helicobacter pylori. AB - The AhpC protein from H. pylori, a thioredoxin (Trx)-dependent alkyl hydroperoxide-reductase, is a member of the ubiquitous 2-Cys peroxiredoxins family (2-Cys Prxs), a group of thiol-specific antioxidant enzymes. Prxs exert the protective antioxidant role in cells through their peroxidase activity, whereby hydrogen peroxide, peroxynitrite and a wide range of organic hydroperoxides (ROOH) are reduced and detoxified (ROOH + 2e(-)-->ROH + H2O). In this study AhpC has been cloned and overexpressed in E. coli. After purification to homogeneity, crystals of the recombinant protein were grown. They diffract to 2.95 A resolution using synchrotron radiation. The crystal structure of AhpC has been determined using the molecular replacement method (R = 23.6%, R(free) = 25.9%). The model, similar in the overall to other members of the 2-Cys Prx family crystallized as toroide-shaped complexes, consists of a pentameric arrangement of homodimers [(alpha2)5 decamer]. The model of AhpC from H. pylori presents significant differences with respect to other members of the family: apart from some loop regions, alpha5-helix and the C-terminus is shifted, preventing the C-terminal tail of the second subunit from extending toward this region of the molecule. Oligomerization properties of AhpC have been also characterized by gel filtration chromatography. PMID- 16213197 TI - Protein kinase structure and function analysis with chemical tools. AB - Protein kinases are the largest enzyme superfamily involved in cell signal transduction and represent therapeutic targets for a range of diseases. There have been intensive efforts from many labs to understand their catalytic mechanisms, discover inhibitors and discern their cellular functions. In this review, we will describe two approaches developed to analyze protein kinases: bisubstrate analog inhibition and phosphonate analog utilization. Both of these methods have been used in combination with the protein semisynthesis method expressed protein ligation to advance our understanding of kinase-substrate interactions and functional elucidation of phosphorylation. Previous work on the nature of the protein kinase mechanism suggests it follows a dissociative transition state. A bisubstrate analog was designed against the insulin receptor kinase to mimic the geometry of a dissociative transition state reaction coordinate distance. This bisubstrate compound proved to be a potent inhibitor against the insulin receptor kinase and occupied both peptide and nucleotide binding sites. Bisubstrate compounds with altered hydrogen bonding potential as well as varying spacers between the adenine and the peptide demonstrate the importance of the original design features. We have also shown that related bisubstrate analogs can be used to potently block serine/threonine kinases including protein kinase A. Since many protein kinases recognize folded protein substrates for efficient phosphorylation, it was advantageous to incorporate the peptide-ATP conjugates into protein structures. Using expressed protein ligation, a Src-ATP conjugate was produced and shown to be a high affinity ligand for the Csk tyrosine kinase. Nonhydrolyzable mimics of phosphoSer/phosphoTyr can be useful in examining the functionality of phosphorylation events. Using expressed protein ligation, we have employed phosphonomethylene phenylalanine and phosphonomethylene alanine to probe the phosphorylation of Tyr and Ser, respectively. These tools have permitted an analysis of the SH2-phosphatases (SHP1 and SHP2), revealing a novel intramolecular stimulation of catalytic activity mediated by the corresponding phosphorylation events. They have also been used to characterize the cellular regulation of the melatonin rhythm enzyme by phosphorylation. PMID- 16213198 TI - Limited proteolysis in the investigation of beta2-microglobulin amyloidogenic and fibrillar states. AB - Amyloid fibrils of patients treated with regular haemodialysis essentially consists of beta2-microglobulin (beta2-m) and its truncated species DeltaN6beta2 m lacking six residues at the amino terminus. The truncated fragment shows a higher propensity to self-aggregate and constitutes an excellent candidate for the analysis of a protein in the amyloidogenic conformation. The surface topology and the conformational analysis of native beta2-m and the truncated DeltaN6beta2 m species both in the soluble and in the fibrillar forms were investigated by the limited proteolysis/mass spectrometry strategy. The conformation in solution of a further truncated mutant DeltaN3beta2-m lacking three residues at the N-terminus was also examined. This approach appeared particularly suited to investigate the regions that are solvent-exposed, or flexible enough to be accessible to protein protein interactions and to describe the conformation of transient intermediates. Moreover, proteolysis experiments can also be tailored to investigate amyloid fibrils by discriminating the protein regions constituting the unaccessible core of the fibrils and those still flexible and exposed to the solvent. Although native beta2-m and DeltaN3beta2-m shared essentially the same conformation, significative structural differences exist between the native and the DeltaN6beta2-m proteins in solution with major differences located at the end moiety of strand V and subsequent loop with strand VI and at both the N- and C termini of the proteins. On the contrary, an identical distribution of preferential proteolytic sites was observed in both proteins in the fibrillar state, which was nearly superimposible to that observed for the soluble form of DeltaN6beta2-m. These data revealed that synthetic fibrils essentially consists of an unaccessible core comprising residues 20-87 of the beta2-m protein with exposed and flexible N- and C-terminal ends. Moreover, proteolytic cleavages observed in vitro at Lys 6 and Lys 19 reproduce specific cleavages that have to take place in vivo to generate the truncated forms of beta2-m occurring in natural fibrils. On the basis of these results, a molecular mechanism for fibril formation has been proposed. PMID- 16213200 TI - Purification and characterization of recombinant Caulobacter crescentus Cu,Zn superoxide dismutase. AB - Recombinant Cu,Zn Superoxide Dismutase from Caulobacter crescentus has been expressed in Escherichia coli and characterized. The corresponding recombinant protein has a molecular weight typical of a homodimeric Cu,ZnSODs and an activity comparable to that of other prokaryotic enzymes. The copper active site is characterized by a peculiar axial geometry as evidenced by its electron paramagnetic resonance spectrum, moreover, the copper atom displays a low accessibility toward external chelating agents indicating a lower solvent accessibility when compared to other prokaryotic enzymes. Investigation of the enzyme thermal stability through differential scanning calorimetry indicates the occurrence of two transitions at low and higher temperature that are found to be due to the apo and holo protein, respectively, confirming that the metals have a crucial role in the stabilization of this class of enzymes. PMID- 16213199 TI - Coupling phosphoryl transfer and substrate interactions in protein kinases. AB - Protein kinases control cell signaling events through the ATP-dependent phosphorylation of serine, threonine and tyrosine residues in protein targets. The recognition of these protein substrates by the kinases relies on two principal factors: proper subcellular co-localization and molecular interactions between the kinase and substrate. In this review, we will focus on the kinetic role of the latter in conveying favorable substrate recognition. Using rapid mixing technologies, we demonstrate that the intrinsic thermodynamic affinities of two protein substrates for their respective kinases (Csk with Src and Sky1p with Npl3) are weak compared to their apparent affinities measured in traditional steady-state kinetic assays (i.e.--Km < Kd). The source of the high apparent affinities rests in a very fast and highly favorable phosphoryl transfer step that serves as a clamp for substrate recognition. In this mechanism, both Csk and Sky1p utilize this step to draw the substrate toward product, thereby, converting a high Kd into a low Km. We propose that this one form of substrate recognition employed by protein kinases is advantageous since it simultaneously facilitates high apparent substrate affinity and fast protein turnover. PMID- 16213201 TI - Perinatal management, counselling and outcome of fetuses with congenital heart disease. AB - Prenatal treatment options for fetal heart disease are still limited but pharmacological treatment of fetal tachyarrhythmias is usually effective. Prenatal catheter interventions are likely to be an option in selected fetal cardiac defects in the future. Delivery should be at a tertiary care centre if the need for immediate neonatal transport is anticipated. When a cardiac problem is diagnosed in a fetus, the parents should be counselled by a paediatric cardiologist specialized in fetal cardiology in close co-operation with the obstetric team. The rate of termination is influenced by gestational age at diagnosis, the severity of the heart defect and the presence of associated malformations. In fetuses with isolated cardiac malformations who are in sinus rhythm with good myocardial function and no or trivial atrioventricular valve regurgitation, the risk of spontaneous intra-uterine death is low. Prenatal echocardiography has the potential to improve postnatal survival in infants with critical heart defects, especially those with duct-dependent systemic or pulmonary circulations. PMID- 16213202 TI - Progression of fetal heart disease and rationale for fetal intracardiac interventions. AB - The outcome of cardiac disease diagnosed before birth is paradoxically worse than that diagnosed postnatally. In part, this is because fetal screening detects cases that are already showing failure of cardiac growth which are usually progressive with secondary damage to the myocardium, lungs and brain. Fetal valvuloplasty has been proposed for cases of critical aortic and pulmonary stenosis or atresia, and atrial septostomy for a restrictive oval foramen associated with aortic stenosis, hypoplastic left heart syndrome and transposition of the great arteries. The rationale for fetal therapy is to restore forward flow and reduce intraventricular pressure, thus improving coronary perfusion and minimizing ischaemic damage. Successful valvuloplasty has reduced systemic venous pressures and reversed fetal hydrops, thus prolonging pregnancy. It has resulted in improved ventricular growth in some cases and spontaneous opening of a closed oval foramen with normalization of pulmonary venous waveforms. These signs suggest better fetal cardiopulmonary development and improved surgical outcomes. PMID- 16213203 TI - Fetal brady- and tachyarrhythmias: new and accepted diagnostic and treatment methods. AB - Sustained bradyarrhythmias are typically the result of symptomatic sinus bradycardia, atrial bigeminy or complete atrioventricular (AV) block. Fetal tachyarrhythmias relate to sinus tachycardia, atrial flutter and supraventricular tachycardia as the main aetiology. Ultrasound is essential to understand the underlying arrhythmia mechanism, to study the impact on cardiac function, to exclude cardiac defects or tumours, and to survey the fetal heart rate and well being, e.g. during anti-arrhythmic treatment. Based on retrospective studies, several more or less safe, effective and well-tolerated anti-arrhythmic agents are currently available for the treatment of atrial and supraventricular tachycardia. Isolated congenital complete AV block is mainly related to maternal anti-Ro/La auto-antibodies. The rationale to treat a fetus at this irreversible stage of AV nodal damage is primarily to mitigate or prevent concomitant myocardial inflammation and to augment cardiac output. A recently published study demonstrated a significant improved outcome with transmaternal dexamethasone and beta-stimulation. PMID- 16213204 TI - Regional distinctions in cortical bone mineral density measured by pQCT can predict alterations in material property at the tibial diaphysis of the Cynomolgus monkey. AB - We examined whether regional differences in cortical bone mineral density (Ct.BMD) measured by peripheral quantitative computed tomography is related to the heterogeneity of bone tissue and whether regional Ct.BMD is a better indicator of changes in bone material properties. Bilateral tibiae were obtained from 17 female adult Cynomolgus monkeys (Macaca fascicularis; mean age 16.8 years). After determining that Ct.BMD was similar between the right and left tibiae, the left tibiae were used for bone histomorphometry and the right for a three-point bending test. The Ct.BMD in the posterior quadrant was significantly higher than that in the anterior quadrant. In the bone histomorphometric analysis, all parameters (i.e., average osteonal area, average osteonal bone area, osteon population density, percent osteonal area [%On.Ar], percent osteonal bone area [%On.B.Ar], percent osteonal area of initial remodeling [%Il.On.Ar], percent osteonal area of secondary remodeling [%Sd.On.Ar], porosity, and percent osteoid area in the posterior region) were significantly lower than those in the anterior region. The results indicated that in the same cross-section, bone tissue structure was heterogeneous. Both total- and posterior-Ct.BMD were positively correlated with breaking stress and negatively correlated with toughness, whereas anterior-Ct.BMD was positively correlated with elastic modulus. Backward stepwise multiple regression analyses indicated that posterior Ct.BMD and total-Ct.BMD were the best variables for predicting breaking stress and toughness, respectively, when age is taken into account. The %On.Ar, %On.B.Ar, and %Il.On.Ar in the posterior region were negatively correlated with elastic modulus. The %On.Ar, %On.B.Ar, and %Sd.On.Ar in the posterior region were positively correlated with toughness. These findings indicated that regional Ct.BMD measurement is useful to assess changes in the material properties of bone associated with the degree of mineralization. In particular, anterior-, posterior , and total-Ct.BMD can be used separately to predict changes in the material properties of the tibial diaphysis. PMID- 16213205 TI - Alternative splicing--when two's a crowd. AB - Various mechanisms have been proposed to explain mutually exclusive splicing of pairs of exons. A paper in this issue of Cell (Graveley, 2005) provides a fascinating insight into the perplexing question of how only one exon at a time is chosen from an array of 48 exons in the Drosophila Dscam gene. PMID- 16213206 TI - A cellular response to an internal energy crisis. AB - Lack of an appropriate energy supply has been thought to induce cell death in a nonspecific manner by causing a decline in metabolism and a gradual cessation of cellular function. In this issue of Cell, Nutt et al. (2005) describe a new mechanism that directly links nutrient availability to apoptosis in Xenopus oocytes and show that age-dependent changes in the nutritional state of a cell might lead to caspase activation and apoptotic cell death. PMID- 16213207 TI - VAV's got rhythm. AB - Biological rhythms with periods of less than a day are physiologically important but poorly understood. In this issue of Cell, Norman, Maricq, and colleagues (Norman et al., 2005) show that VAV-1, a guanine nucleotide exchange factor for Rho-family GTPases, is necessary for three rhythmic behaviors in the nematode Caenorhabditis elegans: feeding, defecation, and ovulation. PMID- 16213208 TI - The blood-brain barrier--lessons from moody flies. AB - Despite the importance of the blood-brain barrier (BBB), little is known about the molecular mechanisms that control its integrity. The identification of moody, a gene required for the formation and maintenance of the Drosophila BBB, provides new insight into how paracellular junctions are formed at the barrier. Meanwhile, moody also has been identified in a screen for fly mutants with altered sensitivity to cocaine, remarkably implicating the BBB in the physiological response to narcotics. PMID- 16213209 TI - Right place, right time, and only once: replication initiation in metazoans. AB - DNA replication is tightly regulated at the initiation step by both the cell cycle machinery and checkpoint pathways. Here, we discuss recent advances in understanding how replication is initiated in metazoans at the correct chromosome positions, at the appropriate time, and only once per cell cycle. PMID- 16213210 TI - Phosphodiesterase 4D deficiency in the ryanodine-receptor complex promotes heart failure and arrhythmias. AB - Phosphodiesterases (PDEs) regulate the local concentration of 3',5' cyclic adenosine monophosphate (cAMP) within cells. cAMP activates the cAMP-dependent protein kinase (PKA). In patients, PDE inhibitors have been linked to heart failure and cardiac arrhythmias, although the mechanisms are not understood. We show that PDE4D gene inactivation in mice results in a progressive cardiomyopathy, accelerated heart failure after myocardial infarction, and cardiac arrhythmias. The phosphodiesterase 4D3 (PDE4D3) was found in the cardiac ryanodine receptor (RyR2)/calcium-release-channel complex (required for excitation-contraction [EC] coupling in heart muscle). PDE4D3 levels in the RyR2 complex were reduced in failing human hearts, contributing to PKA hyperphosphorylated, "leaky" RyR2 channels that promote cardiac dysfunction and arrhythmias. Cardiac arrhythmias and dysfunction associated with PDE4 inhibition or deficiency were suppressed in mice harboring RyR2 that cannot be PKA phosphorylated. These data suggest that reduced PDE4D activity causes defective RyR2-channel function associated with heart failure and arrhythmias. PMID- 16213211 TI - The polyadenylation factor CPSF-73 is involved in histone-pre-mRNA processing. AB - During 3' end processing, histone pre-mRNAs are cleaved 5 nucleotides after a conserved stem loop by an endonuclease dependent on the U7 small nuclear ribonucleoprotein (snRNP). The upstream cleavage product corresponds to the mature histone mRNA, while the downstream product is degraded by a 5'-3' exonuclease, also dependent on the U7 snRNP. To identify the two nuclease activities, we carried out UV-crosslinking studies using both the complete RNA substrate and the downstream cleavage product, each containing a single radioactive phosphate and a phosphorothioate modification at the cleavage site. We detected a protein migrating at 85 kDa that crosslinked to each substrate in a U7-dependent manner. Immunoprecipitation experiments identified this protein as CPSF-73, a known component of the cleavage/polyadenylation machinery. These studies suggest that CPSF-73 is both the endonuclease and 5'-3' exonuclease in histone-pre-mRNA processing and reveal an evolutionary link between 3' end formation of histone mRNAs and polyadenylated mRNAs. PMID- 16213212 TI - Regulation of p53 translation and induction after DNA damage by ribosomal protein L26 and nucleolin. AB - Increases in p53 protein levels after DNA damage have largely been attributed to an increase in the half-life of p53 protein. Here we demonstrate that increased translation of p53 mRNA is also a critical step in the induction of p53 protein in irradiated cells. Ribosomal protein L26 (RPL26) and nucleolin were found to bind to the 5' untranslated region (UTR) of p53 mRNA and to control p53 translation and induction after DNA damage. RPL26 preferentially binds to the 5'UTR after DNA damage, and its overexpression enhances association of p53 mRNA with heavier polysomes, increases the rate of p53 translation, induces G1 cell cycle arrest, and augments irradiation-induced apoptosis. Opposite effects were seen when RPL26 expression was inhibited. In contrast, nucleolin overexpression suppresses p53 translation and induction after DNA damage, whereas nucleolin downregulation promotes p53 expression. These findings demonstrate the importance of increased translation of p53 in DNA-damage responses and suggest critical roles for RPL26 and nucleolin in affecting p53 induction. PMID- 16213213 TI - Mutually exclusive splicing of the insect Dscam pre-mRNA directed by competing intronic RNA secondary structures. AB - Drosophila Dscam encodes 38,016 distinct axon guidance receptors through the mutually exclusive alternative splicing of 95 variable exons. Importantly, known mechanisms that ensure the mutually exclusive splicing of pairs of exons cannot explain this phenomenon in Dscam. I have identified two classes of conserved elements in the Dscam exon 6 cluster, which contains 48 alternative exons--the docking site, located in the intron downstream of constitutive exon 5, and the selector sequences, which are located upstream of each exon 6 variant. Strikingly, each selector sequence is complementary to a portion of the docking site, and this pairing juxtaposes one, and only one, alternative exon to the upstream constitutive exon. The mutually exclusive nature of the docking site:selector sequence interactions suggests that the formation of these competing RNA structures is a central component of the mechanism guaranteeing that only one exon 6 variant is included in each Dscam mRNA. PMID- 16213214 TI - Centriolin anchoring of exocyst and SNARE complexes at the midbody is required for secretory-vesicle-mediated abscission. AB - The terminal step in cytokinesis, called abscission, requires resolution of the membrane connection between two prospective daughter cells. Our previous studies demonstrated that the coiled-coil protein centriolin localized to the midbody during cytokinesis and was required for abscission. Here we show that centriolin interacts with proteins of vesicle-targeting exocyst complexes and vesicle-fusion SNARE complexes. These complexes require centriolin for localization to a unique midbody-ring structure, and disruption of either complex inhibits abscission. Exocyst disruption induces accumulation of v-SNARE-containing vesicles at the midbody ring. In control cells, these v-SNARE vesicles colocalize with a GFP tagged secreted polypeptide. The vesicles move to the midbody ring asymmetrically from one prospective daughter cell; the GFP signal is rapidly lost, suggesting membrane fusion; and subsequently the cell cleaves at the site of vesicle delivery/fusion. We propose that centriolin anchors protein complexes required for vesicle targeting and fusion and integrates membrane-vesicle fusion with abscission. PMID- 16213215 TI - Metabolic regulation of oocyte cell death through the CaMKII-mediated phosphorylation of caspase-2. AB - Vertebrate female reproduction is limited by the oocyte stockpiles acquired during embryonic development. These are gradually depleted over the organism's lifetime through the process of apoptosis. The timer that triggers this cell death is yet to be identified. We used the Xenopus egg/oocyte system to examine the hypothesis that nutrient stores can regulate oocyte viability. We show that pentose-phosphate-pathway generation of NADPH is critical for oocyte survival and that the target of this regulation is caspase-2, previously shown to be required for oocyte death in mice. Pentose-phosphate-pathway-mediated inhibition of cell death was due to the inhibitory phosphorylation of caspase-2 by calcium/calmodulin-dependent protein kinase II (CaMKII). These data suggest that exhaustion of oocyte nutrients, resulting in an inability to generate NADPH, may contribute to ooctye apoptosis. These data also provide unexpected links between oocyte metabolism, CaMKII, and caspase-2. PMID- 16213216 TI - Common molecular pathways mediate long-term potentiation of synaptic excitation and slow synaptic inhibition. AB - Synaptic plasticity, the cellular correlate for learning and memory, involves signaling cascades in the dendritic spine. Extensive studies have shown that long term potentiation (LTP) of the excitatory postsynaptic current (EPSC) through glutamate receptors is induced by activation of N-methyl-D-asparate receptor (NMDA-R)--the coincidence detector--and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Here we report that the same signaling pathway in the postsynaptic CA1 pyramidal neuron also causes LTP of the slow inhibitory postsynaptic current (sIPSC) mediated by metabotropic GABA(B) receptors (GABA(B) Rs) and G protein-activated inwardly rectifying K(+) (GIRK) channels, both residing in dendritic spines as well as shafts. Indicative of intriguing differences in the regulatory mechanisms for excitatory and inhibitory synaptic plasticity, LTP of sIPSC but not EPSC was abolished in mice lacking Nova-2, a neuronal-specific RNA binding protein that is an autoimmune target in paraneoplastic opsoclonus myoclonus ataxia (POMA) patients with latent cancer, reduced inhibitory control of movements, and dementia. PMID- 16213217 TI - The Rho/Rac-family guanine nucleotide exchange factor VAV-1 regulates rhythmic behaviors in C. elegans. AB - Rhythmic behaviors are a fundamental feature of all organisms. Pharyngeal pumping, the defecation cycle, and gonadal-sheath-cell contractions are three well-characterized rhythmic behaviors in the nematode C. elegans. The periodicities of the rhythms range from subsecond (pharynx) to seconds (gonadal sheath) to minutes (defecation). However, the molecular mechanisms underlying these rhythmic behaviors are not well understood. Here, we show that the C. elegans Rho/Rac-family guanine nucleotide exchange factor, VAV-1, which is homologous to the mammalian Vav proto-oncogene, has a crucial role in all three behaviors. vav-1 mutants die as larvae because VAV-1 function is required in the pharynx for synchronous contraction of the musculature. In addition, ovulation and the defecation cycle are abnormal and arrhythmic. We show that Rho/Rac-family GTPases and the signaling molecule inositol triphosphate (IP(3)) act downstream of VAV-1 signaling and that the VAV-1 pathway modulates rhythmic behaviors by dynamically regulating the concentration of intracellular Ca(2+). PMID- 16213218 TI - GPCR signaling is required for blood-brain barrier formation in drosophila. AB - The blood-brain barrier of Drosophila is established by surface glia, which ensheath the nerve cord and insulate it against the potassium-rich hemolymph by forming intercellular septate junctions. The mechanisms underlying the formation of this barrier remain obscure. Here, we show that the G protein-coupled receptor (GPCR) Moody, the G protein subunits G alpha i and G alpha o, and the regulator of G protein signaling Loco are required in the surface glia to achieve effective insulation. Our data suggest that the four proteins act in a complex common pathway. At the cellular level, the components function by regulating the cortical actin and thereby stabilizing the extended morphology of the surface glia, which in turn is necessary for the formation of septate junctions of sufficient length to achieve proper sealing of the nerve cord. Our study demonstrates the importance of morphogenetic regulation in blood-brain barrier development and places GPCR signaling at its core. PMID- 16213219 TI - moody encodes two GPCRs that regulate cocaine behaviors and blood-brain barrier permeability in Drosophila. AB - We identified moody in a genetic screen for Drosophila mutants with altered cocaine sensitivity. Hypomorphic mutations in moody cause an increased sensitivity to cocaine and nicotine exposure. In contrast, sensitivity to the acute intoxicating effects of ethanol is reduced. The moody locus encodes two novel GPCRs, Moody-alpha and Moody-beta. While identical in their membrane spanning domains, the two Moody proteins differ in their long carboxy-terminal domains, which are generated by use of alternative reading frames. Both Moody forms are required for normal cocaine sensitivity, suggesting that they carry out distinct but complementary functions. Moody-alpha and Moody-beta are coexpressed in surface glia that surround the nervous system, where they are actively required to maintain the integrity of the blood-brain barrier in the adult fly. We propose that a Moody-mediated signaling pathway functions in glia to regulate nervous system insulation and drug-related behaviors. PMID- 16213220 TI - Quantitative and dynamic assessment of the contribution of the ER to phagosome formation. AB - Phagosomes were traditionally thought to originate from an invagination and scission of the plasma membrane to form a distinct intracellular vacuole. An alternative model implicating the endoplasmic reticulum (ER) as a major component of nascent and maturing phagosomes was recently proposed (Gagnon et al., 2002). To reconcile these seemingly disparate hypotheses, we used a combination of biochemical, fluorescence imaging, and electron microscopy techniques to quantitatively and dynamically assess the contribution of the plasmalemma and of the ER to phagosome formation and maturation. We could not verify even a transient physical continuity between the ER and the plasma membrane, nor were we able to detect a significant contribution of the ER to forming or maturing phagosomes in either macrophages or dendritic cells. Instead, our data indicate that the plasma membrane is the main constituent of nascent and newly formed phagosomes, which are progressively remodeled by fusion with endosomal and eventually lysosomal compartments as phagosomes mature into acidic, degradative organelles. PMID- 16213221 TI - Telling the liver (not) to make bile acids: a new voice from the gut? AB - The maintenance of adequate amounts of bile acids in the liver, biliary tract, and intestine requires a finely tuned control of their synthesis. A paper in this issue of Cell Metabolism by Inagaki et al. (2005) indicates that sensing of the levels of bile acids in the intestine may trigger the secretion of a hormone which regulates bile acid production in the liver. PMID- 16213222 TI - TORCs rev up gluconeogenesis. AB - Transducer of regulated CREB activity (TORC) proteins promote transactivation by the cAMP response element binding protein (CREB) and mediate effects of cAMP agonists on gene expression. Koo et al. now report that TORC phosphorylation and nuclear/cytoplasmic shuttling play a key role in the regulation of gluconeogenesis by cAMP. Control of TORC phosphorylation and function may integrate the effects of multiple factors involved in metabolic control, including cAMP agonists, insulin, and AMP kinases. TORCs, and kinases affecting TORC function, are promising new therapeutic targets for the treatment of diabetes mellitus. PMID- 16213223 TI - The marriage of glucose and blood vessels: it isn't all that sweet. AB - The secondary complications of diabetes are the major factors limiting quality of life and longevity in diabetic patients. Among these complications, cardiovascular disease is the major cause of morbidity and mortality. Based on a recent paper by Vikramadithyan et al. (2005), this preview addresses the role of aldose reductase as a contributor to the pathogenesis of diabetic macrovascular disease. PMID- 16213224 TI - Fibroblast growth factor 15 functions as an enterohepatic signal to regulate bile acid homeostasis. AB - The liver and intestine play crucial roles in maintaining bile acid homeostasis. Here, we demonstrate that fibroblast growth factor 15 (FGF15) signals from intestine to liver to repress the gene encoding cholesterol 7alpha-hydroxylase (CYP7A1), which catalyzes the first and rate-limiting step in the classical bile acid synthetic pathway. FGF15 expression is stimulated in the small intestine by the nuclear bile acid receptor FXR and represses Cyp7a1 in liver through a mechanism that involves FGF receptor 4 (FGFR4) and the orphan nuclear receptor SHP. Mice lacking FGF15 have increased hepatic CYP7A1 mRNA and protein levels and corresponding increases in CYP7A1 enzyme activity and fecal bile acid excretion. These studies define FGF15 and FGFR4 as components of a gut-liver signaling pathway that synergizes with SHP to regulate bile acid synthesis. PMID- 16213225 TI - The orphan nuclear receptor SHP regulates PGC-1alpha expression and energy production in brown adipocytes. AB - Brown adipocytes increase energy production in response to induction of PGC 1alpha, a dominant regulator of energy metabolism. We have found that the orphan nuclear receptor SHP (NR0B2) is a negative regulator of PGC-1alpha expression in brown adipocytes. Mice lacking SHP show increased basal expression of PGC-1alpha, increased energy expenditure, and resistance to diet-induced obesity. Increased PGC-1alpha expression in SHP null brown adipose tissue is not due to beta adrenergic activation, since it is also observed in primary cultures of SHP(-/-) brown adipocytes that are not exposed to such stimuli. In addition, acute inhibition of SHP expression in cultured wild-type brown adipocytes increases basal PGC-1alpha expression, and SHP overexpression in SHP null brown adipocytes decreases it. The orphan nuclear receptor ERRgamma is expressed in BAT and its transactivation of the PGC-1alpha promoter is potently inhibited by SHP. We conclude that SHP functions as a negative regulator of energy production in BAT. PMID- 16213226 TI - Cdk4 promotes adipogenesis through PPARgamma activation. AB - Cell cycle regulators such as E2F1 and retinoblastoma (RB) play crucial roles in the control of adipogenesis, mostly by controlling the transition between preadipocyte proliferation and adipocyte differentiation. The serine-threonine kinase cyclin-dependent kinase 4 (cdk4) works in a complex with D-type cyclins to phosphorylate RB, mediating the entry of cells into the cell cycle in response to external stimuli. Because cdk4 is an upstream regulator of the E2F-RB pathway, we tested whether cdk4 was a target for new factors that regulate adipogenesis. Here we find that cdk4 inhibition impairs adipocyte differentiation and function. Disruption of cdk4 or activating mutations in cdk4 in primary mouse embryonic fibroblasts results in reduced and increased adipogenic potential, respectively, of these cells. We show that the effects of cdk4 are not limited to the control of differentiation; cdk4 also participates in adipocyte function through activation of PPARgamma. PMID- 16213227 TI - Elevated stearoyl-CoA desaturase-1 expression in skeletal muscle contributes to abnormal fatty acid partitioning in obese humans. AB - Obesity and type 2 diabetes are strongly associated with abnormal lipid metabolism and accumulation of intramyocellular triacylglycerol, but the underlying cause of these perturbations are yet unknown. Herein, we show that the lipogenic gene, stearoyl-CoA desaturase 1 (SCD1), is robustly up-regulated in skeletal muscle from extremely obese humans. High expression and activity of SCD1, an enzyme that catalyzes the synthesis of monounsaturated fatty acids, corresponded with low rates of fatty acid oxidation, increased triacylglycerol synthesis and increased monounsaturation of muscle lipids. Elevated SCD1 expression and abnormal lipid partitioning were retained in primary skeletal myocytes derived from obese compared to lean donors, implying that these traits might be driven by epigenetic and/or heritable mechanisms. Overexpression of human SCD1 in myotubes from lean subjects was sufficient to mimic the obese phenotype. These results suggest that elevated expression of SCD1 in skeletal muscle contributes to abnormal lipid metabolism and progression of obesity. PMID- 16213228 TI - Full intracellular retention of GLUT4 requires AS160 Rab GTPase activating protein. AB - Insulin controls glucose flux into muscle and fat by regulating the trafficking of GLUT4 between the interior and surface of cells. Here, we show that the AS160 Rab GTPase activating protein (GAP) is a negative regulator of basal GLUT4 exocytosis. AS160 knockdown resulted in a partial redistribution of GLUT4 from intracellular compartments to the plasma membrane, a concomitant increase in basal glucose uptake, and a 3-fold increase in basal GLUT4 exocytosis. Reexpression of wild-type AS160 restored normal GLUT4 behavior to the knockdown adipocytes, whereas reexpression of a GAP domain mutant did not revert the phenotype, providing the first direct evidence that AS160 GAP activity is required for basal GLUT4 retention. AS160 is the first protein identified that is specially required for basal GLUT4 retention. Our findings that AS160 knockdown only partially releases basal GLUT4 retention provides evidence that insulin signals to GLUT4 exocytosis by both AS160-dependent and -independent mechanisms. PMID- 16213229 TI - The Pediatric Quality of Life Inventory: an evaluation of its reliability and validity for children with traumatic brain injury. AB - OBJECTIVES: To assess the reliability and validity of the Pediatric Quality of Life Inventory, version 4.0 (PedsQL), and to compare it with that of the Behavior Rating Inventory of Executive Function (BRIEF) among children with traumatic brain injury (TBI). DESIGN: Prospective cohort study that documented the health related quality of life of 391 children at 3 and 12 months postinjury. SETTING: Four level I pediatric trauma centers. PARTICIPANTS: Children (age range, 5-15 y) hospitalized with a TBI or an extremity fracture. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Parent-reported PedsQL and BRIEF scale scores. RESULTS: Both the PedsQL and BRIEF scales showed good internal consistency (PedsQL alpha range, .74-.93; BRIEF alpha range, .82-.98) and test-retest reliability (PedsQL r range, .75-.90; BRIEF r range, .82-.92), respectively. Factor analysis revealed that most PedsQL items loaded most highly on their conceptually derived scale. The PedsQL cognitive function scale detected the largest differences among groups of children with varying severities of TBI as well as parents' assessment of change in cognition postinjury. CONCLUSIONS: Although the reliability of the 2 instruments is comparable, the PedsQL discriminates better among children with TBI. The PedsQL is a promising instrument for measuring the health of children after TBI. PMID- 16213230 TI - Phantom pain, residual limb pain, and back pain in amputees: results of a national survey. AB - OBJECTIVES: To describe the prevalence of amputation-related pain; to ascertain the intensity and affective quality of phantom pain, residual limb pain, back pain, and nonamputated limb pain; and to identify the role that demographics, amputation-related factors, and depressed mood may contribute to the experience of pain in the amputee. DESIGN: Cross-sectional survey. SETTING: A sample of persons who contacted the Amputee Coalition of America from 1998 to 2000 were interviewed by telephone. PARTICIPANTS: A stratified sample by etiology of 914 persons with limb loss. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Prevalence, intensity, and bothersomeness of residual, phantom, and back pain, depressed mood as measured by the Center for Epidemiologic Study Depression Scale, characteristics of the amputation, prosthetic use, and sociodemographic characteristics of the amputee. RESULTS: Nearly all (95%) amputees surveyed reported experiencing 1 or more types of amputation-related pain in the previous 4 weeks. Phantom pain was reported most often (79.9%), with 67.7% reporting residual limb pain and 62.3% back pain. A large proportion of persons with phantom pain and stump pain reported experiencing severe pain (rating 7-10). Across all pain types, a quarter of those with pain reported their pain to be extremely bothersome. Identifiable risk factors for intensity and bothersomeness of amputation-related pain varied greatly by pain site. However, across all pain types, depressive symptoms were found to be a significant predictor of level of pain intensity and bothersomeness. CONCLUSIONS: Chronic pain is highly prevalent among persons with limb loss, regardless of time since amputation. A common predictor of an increased level of intensity and bothersomeness among all pain sites was the presence of depressive symptoms. Further studies are needed to elucidate the relationship between pain and depressive symptoms among amputees. PMID- 16213231 TI - Inpatient rehabilitation following burn injury: patient demographics and functional outcomes. AB - OBJECTIVE: To identify demographic characteristics of burn patients referred for inpatient rehabilitation, the benefits of rehabilitation in this population, and factors that influence functional outcome. DESIGN: Retrospective chart review. SETTING: Free-standing rehabilitation hospital. PARTICIPANTS: Patients (N=129) admitted for inpatient rehabilitation after a burn injury. INTERVENTION: Comprehensive inpatient rehabilitation. MAIN OUTCOME MEASURES: Demographic data and the FIM instrument on admission and discharge for all patients. RESULTS: Linear measures of functional status derived by Rasch analysis of the FIM showed significant improvements from admission to discharge for all patients. There was no correlation between total body surface area (TBSA), premorbid psychiatric alcohol or drug abuse history, and change between admission and discharge FIM score. There was a significant correlation between TBSA burn and age and between TBSA and length of stay. CONCLUSIONS: Burn patients referred for inpatient rehabilitation are either older or have large TBSA burns. All patients made significant functional improvements and consequently inpatient rehabilitation can be considered an important component of care after a burn injury. PMID- 16213232 TI - Benefits of cardiac rehabilitation in patients with implantable cardioverter defibrillators: a patient survey. AB - OBJECTIVES: To determine whether participation in an outpatient cardiac rehabilitation (OCR) program increases exercise levels and decreases shocks in patients with implantable cardioverter-defibrillators (ICDs). DESIGN: Retrospective comparative survey. SETTING: University tertiary-care ICD clinic. PARTICIPANTS: Patients with ICDs and coronary artery disease. INTERVENTION: Participation in OCR (nonrandomized). MAIN OUTCOME MEASURES: Exercise levels and OCR participation were determined through a telephone survey of patients with ICDs. The incidence of shock-treated arrhythmia was determined by review of charts and ICD data storage disks. RESULTS: Of 82 patients (85% men; mean age, 61+/-8 y), 28 (34%) participated in OCR after receiving an ICD. There was no difference in age, sex, ejection fraction, or length of follow-up between OCR and non-OCR groups. Median intensity of regular exercise was 5.3 metabolic equivalents (METS) for OCR patients versus 3.5 METS for non-OCR patients (P<.02). In follow-up (mean, 48+/-3 mo), non-OCR patients were more likely to receive any shock, shocks during exercise, or shocks for ventricular arrhythmia during exercise than OCR patients (all P<.05). Non-OCR remained a predictor of shock after adjustment for exercise limitation (P<.05). CONCLUSIONS: OCR patients exercised more and had fewer shocks. Physicians and health plans should encourage ICD patients to participate in OCR. PMID- 16213233 TI - Application of a virtual reality-enhanced exercise protocol in patients after coronary bypass. AB - OBJECTIVE: To determine whether, by using training enhanced by virtual reality (VR) displays, patients who had undergone coronary artery bypass graft (CABG) surgery could more effectively attain long-term cardiopulmonary results than those not using VR technology. DESIGN: Randomized controlled trial. SETTING: Exercise testing laboratory in a medical center. PARTICIPANTS: Consecutive sample of 32 patients who had undergone CABG surgery. INTERVENTIONS: All subjects were randomly assigned to 1 of 2 submaximal endurance programs, with or without simulated training. In all other respects, the 2 programs were identical. Each section lasted for 30 minutes and was conducted twice a week for about 3 months. Graded exercise tests, conducted before and after training, evaluated cardiorespiratory changes. MAIN OUTCOME MEASURES: The cardiorespiratory and hemodynamic parameters were evaluated at both peak and submaximal exertion. RESULTS: At the follow-up exercise tests (performed at 5.4 mo after surgical intervention), the VR group achieved significantly higher value in peak oxygen consumption (VO2peak), peak metabolic equivalents (METS), and amount of VO2 at anaerobic threshold than the non-VR group. Significant gain was attained in VO2peak and peak MET value accrued to those who participated in simulation-based aerobic training. CONCLUSIONS: Our study outcomes clearly support the perceived benefits of rehabilitation programs that incorporate VR to augment patients' recovery of their physical capacity. PMID- 16213234 TI - Exploring the content of physiotherapeutic home-based stroke rehabilitation in New Zealand. AB - OBJECTIVE: To address the paucity of information on the content of home interventions for people with stroke by reporting on the practice of physiotherapeutic home-based stroke rehabilitation in New Zealand. DESIGN: Qualitative research methodology comprising a series of semi-structured interviews. SETTING: Community setting in 6 cities in New Zealand. PARTICIPANTS: A purposeful sampling strategy recruited 20 physiotherapists working in home based stroke rehabilitation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Not applicable. RESULTS: Participants described patients as being fatigued, frustrated, depressed, and scared once discharged home and said that the primary aim of rehabilitation in the home environment is preparation for life after stroke. Physiotherapists aimed at optimal independent functioning by building patients' confidence, self-responsibility, and problem-solving skills while ensuring patient safety. Participants, illustrating the complexities of stroke rehabilitation, described a wide range of interventions. We identified a number of factors that influenced the practice decisions made by participants. The success of intervention was measured more by the successful attainment of carefully set patient-centered goals than by the use of validated outcome measures. CONCLUSIONS: This study presents a conceptual model or framework for physiotherapy practice for people with stroke living in the community. PMID- 16213235 TI - Cost-effectiveness of screening for deep vein thrombosis by ultrasound at admission to stroke rehabilitation. AB - OBJECTIVE: This project was a cost-effectiveness analysis of the practice of routine Doppler ultrasound screening for deep vein thrombosis (DVT) in patients with ischemic stroke at the time of admission to rehabilitation. DESIGN: A decision-analysis model was created to compare 2 approaches for detecting DVT in a stroke population: (1) screening all patients with acute ischemic stroke at admission to stroke rehabilitation for DVT by Doppler ultrasound with subsequent treatment; or (2) clinical surveillance for signs of DVT and treatment after confirmation by Doppler ultrasound. The prevalence of DVT, risk of complication from DVT, and risk of complication from treatment were obtained from published reports. Costs are in 2004 dollars and the effectiveness was measured in quality adjusted life-years (QALYs) gained. We conducted these analyses from a societal perspective. SETTING: Inpatient stroke rehabilitation unit. PARTICIPANTS: Not applicable. INTERVENTION: Screening all patients with acute ischemic stroke at admission to stroke rehabilitation for DVT by Doppler ultrasound with subsequent treatment. MAIN OUTCOME MEASURE: Cost in 2004 dollars per QALY gained by screening all patients with acute ischemic stroke at admission to stroke rehabilitation for DVT by Doppler ultrasound with subsequent treatment. RESULTS: The expected utility of screening patients with ischemic stroke for DVT by Doppler ultrasound on admission to rehabilitation is 1.875 QALYs and that of not screening is 1.872 QALYs. The expected gain is .0026 QALYs (23 h). Obtaining this increase in quality-adjusted life incurs additional cost of 168 dollars per stroke patient and a marginal cost-effectiveness of 67,200 dollars for each QALY gained. CONCLUSIONS: This study estimates that the cost-effectiveness ratio is considerably higher than that reported in other rehabilitation conditions and higher than the commonly stated level for an intervention to be considered cost effective. The difference from previous reports primarily relates to the shorter life expectancy following stroke, the prevalence of occult DVT at admission, rate of complications of anticoagulation, and the estimates of the screening's test characteristics used in our study. Further study of these areas is likely to contribute to improving our understanding of the most appropriate care of these patients. PMID- 16213236 TI - Inpatient rehabilitation of patients requiring hemodialysis: improving efficiency of care. AB - OBJECTIVE: To determine if reducing missed therapy sessions by patients requiring hemodialysis will reduce the length of stay (LOS) and improve the efficiency of care in an inpatient rehabilitation unit. DESIGN: Retrospective study. SETTING: Inpatient rehabilitation unit at a university medical center. PARTICIPANTS: All patients discharged from the Albany Medical Center rehabilitation unit between January 1, 2003, and June 30, 2004. The total number of patients was 955, 40 of whom required hemodialysis. INTERVENTION: On January 1, 2003, Albany Medical Center began providing hemodialysis in the late afternoon, allowing patients to complete a full 3-hour program without missing therapy sessions to attend dialysis. We compared the outcomes of 915 patients who did not require hemodialysis with the outcomes of 40 patients who required hemodialysis. We also compared the outcomes of patients treated on the rehabilitation unit in 2003-2004 to the outcomes of patients treated on the rehabilitation unit before dialysis was available at times that did not conflict with therapy (calendar year 2001). MAIN OUTCOME MEASURES: LOS, improvement on the FIM instrument, and FIM efficiency score. RESULTS: The LOS of the hemodialysis patients changed from 16.0 in 2001 to 12.1 in 2003-2004 (P<.05), and the FIM efficiency score of the hemodialysis patients improved from 1.8 in 2001 to 2.9 in 2003-2004 (P<.05). The FIM efficiency score of the nondialysis group in 2003-2004 was 4.0. This was significantly greater (P<.05) than that of dialysis patients in 2003-2004. CONCLUSIONS: A program to reduce conflicts between medical treatments such as hemodialysis and therapy sessions may result in reduced LOS and improved efficiency on an inpatient rehabilitation unit. PMID- 16213237 TI - Effects on function and quality of life of postoperative home-based physical therapy for patients with hip fracture. AB - OBJECTIVE: To evaluate the effects of a 3-month home-based physical therapy (PT) program for patients with hip fracture after surgery. DESIGN: Randomized controlled trial. SETTING: Home. PARTICIPANTS: Twenty-five patients recently discharged from an acute orthopedic department. INTERVENTIONS: Patients were randomized to the home-based PT group (n=13), where they received home-based PT 8 times from discharge to month 3 postdischarge, or to the control group (n=12). The home-based PT program included exercises for muscle strengthening, range of motion (ROM), balance, and functional training. Patients in the control group were instructed to practice the exercise program given at bedside before discharge. MAIN OUTCOME MEASURES: Patients were evaluated for hip ROM, strength, walking velocity, Harris hip score, and health-related quality of life (HRQOL) at the week of discharge and at 1, 3, and 6 months after discharge. RESULTS: The baseline characteristics showed no difference between the 2 groups. Harris score of the home-based PT group progressed from 58.6+/-8.5 to 90.1+/-5.4 at month 3, whereas Harris score of the control group progressed from 54.6+/-14.5 to 77.4+/ 10.0 (P<.01). Scores of the psychologic domain of HRQOL for the home-based PT group were significantly better at month 1 (P<.05) and month 3 (P<.01) after discharge. Moreover, the physical domain score of the home-based PT group was also significantly better (P<.05) at 3 months after discharge. CONCLUSIONS: Home based PT programs could help patients regain function and HRQOL earlier. PMID- 16213238 TI - Altering asymmetric limb loading after hip arthroplasty using real-time dynamic feedback when walking. AB - OBJECTIVE: To evaluate a walking program incorporating real-time biofeedback to reduce asymmetric limb loading after total hip arthroplasty (THA). DESIGN: Within subject clinical intervention. SETTING: Biomechanics laboratory. PARTICIPANTS: Volunteers were screened for confounding disorders that could affect their gait other than unilateral THA. Participants included 28 subjects who were evaluated a minimum of 2 months after surgery and ambulatory without assistive devices. INTERVENTIONS: THA subjects were assigned to a feedback, no-feedback, or control group. The feedback group walked on a treadmill 15 minutes, 3 times a week for 8 weeks while matching step-to-step reaction forces. Subjects walking without feedback had equal time. The control group did not train. MAIN OUTCOME MEASURES: Symmetry indices for peak limb-loading force, rate of rise of loading force, and impulse calculated from vertical foot-ground forces. Symmetry index changes were evaluated using 2-factor, repeated-measures analyses of variance with a Tukey post hoc test. RESULTS: Loading rate and impulse equalization improved for the feedback group (P<.01). Loading rate equalization improved for the no-feedback group (P=.01). There were no changes for the control group. CONCLUSIONS: This preliminary study suggests that a treadmill walking program may help persons with a THA achieve a more symmetric gait. Additional investigation of the potential benefits of a rehabilitation program incorporating treadmill walking with and without biofeedback is recommended. PMID- 16213239 TI - Fasting serum levels of adiponectin, ghrelin, and leptin in men with spinal cord injury. AB - OBJECTIVES: To measure serum levels of adiponectin, ghrelin, and leptin in men with spinal cord injury (SCI) and to investigate possible correlations between these serum levels and various factors, such as body mass index (BMI), age, injury level, and duration of injury. DESIGN: Cross-sectional. SETTING: A university hospital that is a tertiary referral center. PARTICIPANTS: Eighty-nine men with traumatic neurologically complete SCI (30 with tetraplegia, 59 with paraplegia) and 37 age- and BMI-matched male controls. Subjects with SCI were injured at the mean age +/- standard error of 28.5+/-1.0 years (range, 14.7-59.1 y) and the mean injury duration was 10.8+/-0.7 years (range, 1.1-27.7 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Serum levels of adiponectin, ghrelin, and leptin and BMI. RESULTS: Serum leptin levels in subjects with SCI (mean, 7.0+/-0.5 mg/mL) [corrected] were significantly higher than those in able-bodied controls (mean, 4.7+/-0.6 mg/mL) [corrected] (P<.01). The group with tetraplegia had higher serum leptin levels than the group with paraplegia, but this did not reach a statistically significant level (8.2+/-1.1 ng/mL vs 6.4+/-0.5 mg/mL [corrected] P=.097). There were significant differences in serum leptin levels among the 3 groups by 1-way analysis of variance (P=.008). Serum adiponectin levels in subjects with SCI (7.1+/-0.5 mg/mL) [corrected] were higher than those in able-bodied controls (5.6+/-0.5 mg/mL) [corrected] but this was not statistically significant (P=.08). In contrast, serum levels of ghrelin in subjects with SCI (302.0+/-17.5 pg/mL) were similar to those in the controls (264.0+/-27.0 pg/mL) (P=.24). Serum leptin levels correlated positively with BMI (SCI, r=.698, P<.001; controls, r=.782, P<.001), whereas serum adiponectin (SCI, r=-.527, P<.001; controls, r=-.315, P=.057) and ghrelin (SCI, r=-.368, P<.001; controls, r=-.447, P=.006) levels correlated negatively with the BMI in both subjects with SCI and controls. CONCLUSIONS: Men with SCI have significantly higher serum leptin levels than able-bodied controls, and serum leptin levels correlated with the degree of neurologic deficit. Men with SCI had a tendency toward higher serum adiponectin level than able-bodied controls. Serum levels of ghrelin in men with SCI were similar to those of controls. PMID- 16213240 TI - Bone mineral density after spinal cord injury: a reliable method for knee measurement. AB - OBJECTIVES: To test the interrater reliability of a standardized method to analyze knee bone mineral density (BMD) using dual-energy x-ray absorptiometry (DXA); to compare spine, hip, and knee BMD of people with spinal cord injury (SCI) with able-bodied controls; and to determine the relation between hip BMD and knee BMD in SCI and able-bodied subjects. DESIGN: Criterion standard and masked comparison. SETTING: Primary care university hospital. PARTICIPANTS: A convenience sample of 11 subjects with complete SCI was age and sex matched with 11 able-bodied control subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Four raters analyzed regions of interest according to operational definitions recently developed to standardize the analysis of BMD of the knee. Subjects with chronic SCI and matched controls underwent conventional DXA scans of the spine and hips and "less conventional" scans of the distal femurs and proximal tibias. The relation between hip and knee BMD was analyzed. RESULTS: The knee measurements were highly reliable (femur intraclass correlation coefficient model 2,1 [ICC(2,1)]=.98; tibia ICC(2,1)=.89). Subjects with SCI had lower BMD values than controls at all hip and knee sites (P<.05). Lumbar spine BMD did not differ between groups. Hip BMD was moderately predictive of distal femur BMD (R2=.67), but less correlated with the proximal tibia (R2=.38). CONCLUSIONS: Knee BMD can be reliably analyzed using DXA with this protocol. Subjects with SCI have diminished knee and hip BMD. Low hip BMD is associated with low distal femur BMD. PMID- 16213241 TI - Respiratory pressures and thoracoabdominal motion in community-dwelling chronic stroke survivors. AB - OBJECTIVE: To investigate and characterize the muscular strength and components of breathing patterns in chronic stroke survivors compared with age-matched subjects. DESIGN: Descriptive case-control study. SETTING: Research laboratory. PARTICIPANTS: Sixteen community-dwelling stroke survivors and 19 age-matched healthy subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Maximal inspiratory (MIP) and expiratory pressure (MEP), tidal volume, respiratory rate, percentage of inspiratory time related to total respiratory cycle, mean inspiratory flow, and rib cage and abdominal contributions to tidal volume. RESULTS: Stroke subjects presented significantly lower MIP (P<.01) and MEP (P=.01) and a tendency for predominance of rib cage contributions (P=.05) during tidal breathing. There were no significant differences between the other respiratory variables. CONCLUSIONS: Decreases in strength of respiratory muscles and lower abdomen contributions were shown during the respiratory cycle in community-dwelling stroke subjects. Based on these findings, it can be hypothesized that the association of specific respiratory training could optimize the gains associated with cardiovascular training in stroke survivors. PMID- 16213242 TI - Activity monitoring for assessment of physical activities in daily life in patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: To investigate the degree of agreement between different methods of assessing physical activities in daily life in patients with chronic obstructive pulmonary disease (COPD): video recordings (criterion standard), the DynaPort Activity Monitor (DAM), and patient self-report. DESIGN: Study A: outcomes from video recordings were compared with DAM outcomes and with patient estimation of time spent on each activity after a 1-hour protocol including walking, cycling, standing, sitting, and lying. Study B: DAM outcomes and patient self-report were compared during 1 day in real life. SETTING: Outpatient clinic in a university hospital. PARTICIPANTS: Study A: 10 patients with COPD (mean age, 62+/-6 y; forced expiratory volume in the first second [FEV1]=40%+/-16% of predicted). Study B: 13 patients with COPD (mean age, 61+/-8 y; FEV1=33%+/-10% of predicted). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Time spent on different activities and movement intensity during walking and cycling. RESULTS: Study A: time estimated by the patients in the sitting position was significantly lower than the time showed by the video recordings and the DAM (both P<.001). For the other variables, there were no statistically significant differences (all P>.05). However, Bland and Altman plots and intraclass correlation coefficients showed large disagreement between video recordings and patients' estimations, in contrast to the high degree of agreement between video recordings and DAM. Changes in walking speed correlated highly to changes in DAM movement intensity (r=.81, P<.01). Study B: patients significantly overestimated walking time (22+/ 47 min, P=.04) and underestimated standing time (-45+/-71 min, P=.04). CONCLUSIONS: The DAM showed high accuracy in objectively assessing time spent on different activities and changes in walking speed in patients with COPD. Patients' estimations of time spent on physical activities in daily life disagreed with objective assessment. PMID- 16213243 TI - Influence of mastication and salivation on swallowing in stroke patients. AB - OBJECTIVE: To determine differences of the oral swallowing functions between stroke and normal subjects and to identify those factors affecting dysphagia of stroke patients. DESIGN: Case-control study. SETTING: University hospital. PARTICIPANTS: Ten stroke and 10 healthy subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Resting saliva flow rate, the total number of chews, and the duration of the oral phase until onset of pharyngeal swallow, and food viscosities both before and after the oral phase using diluted barium, pudding, thick rice gruel, and curd type yogurt. RESULTS: The viscosity of all test foods was significantly reduced after the oral phase in both groups (P<.01). Resting saliva flow rate and the viscosity of thick rice gruel after the oral phase in the stroke group were significantly lower than in the control group (P<.01). However, there were no differences in the viscosities of the other foods between both groups. And a longer oral phase and a greater number of chews prior to pharyngeal swallow were revealed in the stroke group than in the control group (P<.01). CONCLUSIONS: This study shows that the food requiring mastication demands a longer oral phase and a greater number of chews, and has more altered rheologic characteristics during the oral phase in stroke patients. These findings suggest that masticatory function is impaired in stroke patients, which may contribute to their swallowing dysfunction. PMID- 16213244 TI - Development of a new method for objective assessment of spasticity using full range passive movements. AB - OBJECTIVE: To develop a method for assessment of spasticity, in which the whole range of motion (ROM) at a wide variation of speeds is applied. DESIGN: Cross sectional design to study construct validity. SETTING: Research department affiliated with a rehabilitation hospital in The Netherlands. PARTICIPANTS: Nine patients with complete spinal cord injury recruited from the rehabilitation hospital. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Thirty to 45 stretches over the whole ROM were applied to the triceps surae muscle at varying velocities measuring from 30 degrees to 150 degrees/s. Electromyographic responses were measured in order to assess reflex excitability. The torque over the ankle joint was measured during the whole stretch. The angle and velocity at which the reflex was initiated was also determined. RESULTS: The electromyographic responses increased significantly at increasing stretch velocities (P<.001). The applied maximum angles are reproducible (intraclass correlation coefficient, .81) and provide representative torque responses. CONCLUSIONS: The assessment method of spasticity using full range passive movements provides objective outcomes. The angular velocity is responsible for an exponential increase in amplitude of the electromyographic response. PMID- 16213245 TI - Reliability and validity of static knee strength measurements obtained with a chair-fixed dynamometer in subjects with hip or knee arthroplasty. AB - OBJECTIVES: To evaluate the reliability (intertrial, interevaluator) and the concurrent validity of strength measurements obtained with a chair-fixed dynamometer and to recommend a clinical protocol that minimizes standard error of measurement (SEM). DESIGN: Within-session repeated measures of maximal static strength of knee flexors and extensors at 30 degrees and 60 degrees of flexion on the chair-fixed and Cybex dynamometers. SETTING: Ambulatory physiotherapy department of a rehabilitation hospital. PARTICIPANTS: Convenience sample of 50 subjects with total hip (n=25) or knee (n=25) arthroplasty. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Reliability was quantified by indices of dependability and corresponding SEMs estimated with the generalizability theory, whereas coefficients of correlation were used to assess the validity. RESULTS: Indices of dependability confirmed excellent intertrial (0.98-1.00) and a very good interevaluator (.92-.99) reliability for the strength measures obtained for different movements and positions. An average of 3 trials minimized the magnitude of the SEMs (>2 Nm for all measurements). When comparing the strength values obtained with the Cybex dynamometer to those measured with the chair-fixed dynamometer, strongest relations were attained when the tested knee was positioned at 60 degrees compared with 30 degrees for subjects with a total knee (.78-.92 vs .87-.93) or hip (.73-.85 vs .86-.91) arthroplasty. CONCLUSIONS: A clinical protocol averaging 3 trials with the knee positioned at 60 degrees after a familiarization period, both for knee flexors and extensors, performed by a trained therapist is recommended to minimize measurement errors on strength values measured with the chair-fixed dynamometer. PMID- 16213246 TI - Endurance times of trunk muscles in male intercollegiate rowers in Hong Kong. AB - OBJECTIVES: To establish isometric endurance times of trunk muscles and their ratios in a group of healthy intercollegiate rowers in Hong Kong for clinical assessment reference, and to compare the trunk endurance profile of the rowers in the current study with that of nonrowers in another study. DESIGN: Isometric endurance times were measured in 4 different positions in a cross-sectional manner. A subset of 5 subjects was tested 3 times 2 days and 1 week apart to evaluate reliability. SETTING: Sports medicine department of a national sports institute. PARTICIPANTS: Thirty-two subjects selected from a group of 42 male intercollegiate rowers reported to have more than 6 months of rowing experience and without history of low back pain. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Trunk muscle endurance times in seconds and ratios of endurance times normalized to that of the extensor muscle. RESULTS: The trunk flexor (mean +/- standard deviation, 176.56+/-88.58 s) had the best endurance times among all the trunk muscles tested (extensor mean, 114.28+/-34.62 s; left lateral flexor mean, 94.53+/-32.97 s; right lateral flexor mean, 98.13+/-41.38 s). No significant difference was found between the left and right lateral flexors (P<.05). The lateral flexor and the flexor endurance times were 85% and 154% of that of the extensor, respectively. The testing protocol in this group of rowers showed good to excellent reliability (intraclass correlation coefficient range, .76-.93). CONCLUSIONS: Intercollegiate rowers in Hong Kong have better endurance in their trunk flexor than the extensor; the lateral flexors are of similar endurance capacity. These findings are different from the endurance profile reported for nonrowers in a previous study. Such differences should be considered when evaluating trunk endurance times in rowers for rehabilitation and training. PMID- 16213247 TI - Nonuniform activity of human calf muscles during an exercise task. AB - OBJECTIVES: To determine the distribution of leg muscle activity during heel raises using magnetic resonance imaging (MRI) with special emphasis on quantifying activity across multiple axial sections and to determine if there are differences among portions of active muscles. DESIGN: Pre- and postexercise (heel raise) T2-weighted time measurements were assessed by using repeated-measures analysis of variance (ANOVA) and t tests. SETTING: Laboratory and MRI suites. PARTICIPANTS: Eight healthy volunteers. INTERVENTION: Unilateral heel raises every 2 seconds for at least 60 seconds. MAIN OUTCOME MEASURES: Percentage changes from T2-weighted magnetic resonance images of the lateral gastrocnemius, medial gastrocnemius, peroneus longus, soleus, and tibialis anterior muscles, across 10 axial sections, exercise bouts, and a pre-exercise condition. RESULTS: The lateral gastrocnemius, medial gastrocnemius, peroneus longus, and soleus had significantly larger changes in T2 time from pre-exercise times than did the tibialis anterior for whole muscles as determined by using repeated-measures ANOVA and post hoc analyses. The medial gastrocnemius had a significantly greater change in T2 time than the lateral gastrocnemius. Proximal axial sections of the lateral gastrocnemius, medial gastrocnemius, and soleus had significantly larger changes in T2 time from pre-exercise than did distal sections. CONCLUSIONS: This work reconfirms that multiple muscles contribute to plantarflexor forces and additionally shows an apparent proximal versus subvolume organization of activity within the gastrocnemius, medial gastrocnemius, and soleus but not the peroneus longus. This proximal versus distal organization of muscle activity needs further investigation. There may be clinical implications for therapeutic interventions that require accurate placement of electrodes such as biofeedback. PMID- 16213248 TI - Modulation of the sleep state-dependent P50 midlatency auditory-evoked potential by electric stimulation of acupuncture points. AB - OBJECTIVE: To determine if the P50 midlatency auditory evoked potential, a sleep state-dependent waveform thought to be generated by the reticular activating system, is modulated after surface stimulation of acupuncture points (ie, electroacupuncture). DESIGN: P50 potential recordings were carried out before, during, and after electroacupuncture. SETTING: A clinical research center. PARTICIPANTS: Eighty healthy subjects ages 25 to 55 were recorded in 7 investigations. INTERVENTIONS: Stimulation of 3 specific acupuncture points (Pericardium 6, Heart 3, Liver 3) was compared with no stimulation or with stimulation of control points (Gall Bladder 34, Large Intestine 11, Small Intestine 3). We compared different frequencies of stimulation (5, 60, 100 Hz), unilateral versus bilateral stimulation, and the effects of repeated episodes of stimulation. MAIN OUTCOME MEASURES: P50 auditory evoked potential latency, amplitude (measure of level of arousal), and habituation (measure of sensory gait) at interstimulus interval of 250 ms. RESULTS: Electroacupuncture at specific points decreased P50 potential amplitude versus electroacupuncture at control points (P=.006) or versus no stimulation (P<.001). The optimal effective frequency was 5 Hz (P<.05 at 5 Hz, P>.05 at 60 and 100 Hz), and unilateral electroacupuncture was not as effective as bilateral electroacupuncture (P=.007). Repeated episodes of bilateral electroacupuncture showed additive effects (P<.05). There were no differences in responsiveness across sexes (P=.79), and electroacupuncture did not affect P50 potential habituation (P>.05). CONCLUSIONS: Electroacupuncture may be effectively used to decrease arousal levels, perhaps as adjunct therapy for disorders of hypervigilance. PMID- 16213249 TI - Motor strategies responsible for maintaining standing posture after deafferentation of the unilateral leg. AB - OBJECTIVE: To identify the motor strategies responsible for maintaining standing posture after deafferentation of the unilateral leg. DESIGN: Pretest-posttest, repeated-measures design. SETTING: A Japanese university hospital rehabilitation facility. PARTICIPANTS: Nine healthy subjects aged 25 to 32 years. INTERVENTION: Two separate sessions that consisted of prolonged standing (control task) and standing until the H-reflex disappeared through the application of an inflated pneumatic cuff above the right knee. MAIN OUTCOME MEASURES: Center of pressure (COP), hip and ankle joint positions, and electromyographic activities. RESULTS: In the control task, no time-related change during prolonged standing was found in any of the measured parameters. After deafferentation, the right soleus activity decreased significantly (P<.01), so the mean velocities of the COP in the anteroposterior and mediolateral directions were greater and the average COP position under the right foot shifted backward compared with those in the previous periods (P<.01). Also, the left tibialis anterior and soleus were activated, as was the bilateral gluteus medius. CONCLUSIONS: The unilateral loss of leg and foot sensory information necessitated additional regulatory activities to stabilize the standing posture. The newly organized posture appeared to partly simulate the standing posture in patients with sensory disturbance of a unilateral leg. PMID- 16213250 TI - Reliability of traditional and fractal dimension measures of quiet stance center of pressure in young, healthy people. AB - OBJECTIVES: To assess reliability of traditional and fractal dimension measures of quiet stance center of pressure (COP). DESIGN: Cross-sectional study. SETTING: University laboratory. PARTICIPANTS: Thirty young healthy men (n=20) and women (n=10) (mean age, 23 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: COP was recorded for 3 trials across 4 conditions: eyes open and eyes closed standing on firm and foam surfaces. Traditional COP variables--peak sway velocity and range of sway, both in the anteroposterior (AP) and mediolateral (ML) directions, and total excursion area, and fractal dimension of the COP in the AP and ML directions--were calculated. Reliability statistics were calculated. RESULTS: Range of sway (AP) was the most reliable traditional variable (intraclass correlation coefficient model 2,1 [ICC(2,1)] range -.28 to .72.). Peak sway velocity (AP) had poorest reliability (ICC(2,1) range, .05-.29). Only 1 of the traditional variables had excellent reliability; total excursion area (firm, eyes closed) (ICC(2,1)=.95). All bar 1 fractal dimension measures had excellent ICCs. Relative technical error of measurement ranged from 4% to 7% for the fractal dimension measures. Coefficients of variation were also very good, ranging from 1.8% to 6.7%. CONCLUSIONS: Fractal dimension measures were more reliable than traditional measures of COP. Although traditional measures are used extensively to assess COP, their reliability is questionable. Fractal dimension measures show promise to reliably quantify COP and warrant further investigation. PMID- 16213251 TI - The reliability and validity of the physiological cost index in healthy subjects while walking on 2 different tracks. AB - OBJECTIVE: To investigate the reliability and validity of the Physiological Cost Index (PCI) scores, as a measure of energy expenditure, when healthy subjects walk on 2 different tracks (20-m and 12-m figure eight tracks). DESIGN: Intra- and interrater reliability and construct validity. SETTING: Physiotherapy division of a university in London, UK. PARTICIPANTS: Forty healthy subjects (15 men, 25 women; mean age +/- standard deviation, 34.5+/-12.6 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Heart rate (in beats/min) and speed (in m/min) were used to calculate the PCI (in beats/m). Rate of oxygen consumption (VO2, in mL x kg(-1) x min(-1)) and oxygen cost (EO2, in mL x kg(-1) x m(-1)) were used as criterion estimates of energy cost EO2. Pearson correlation coefficients between the PCI, components of the PCI, EO2, and VO2 were used to quantify validity. Intrarater reliability was assessed in all participants and interrater reliability was assessed on a subset of 13 subjects using intraclass correlation coefficients and Bland-Altman plots. RESULTS: Intrarater (r=.73, r=.79) and interrater (r=.62, r=.66) reliability were acceptable between PCI scores from 20-m and 12-m tracks, respectively. Correlations between VO2 and EO2 with PCI were weak. PCI scores from the 20-m track were significantly lower than those on the 12-m track (P=.002). Subjects walked significantly faster on the 20 m track (P<.001). Results suggest a large difference in PCI scores would be necessary to indicate a "true" alteration in performance (52% for 20-m track, 43.4% for the 12-m track). CONCLUSIONS: The PCI is reliable but not valid as a measure of the energy cost of walking in healthy subjects, on either track. The 20-m track is recommended for clinical use because it enables subjects to walk at a faster pace. PMID- 16213252 TI - Interexaminer repeatability of antidromic ulnar sensory nerve conduction velocity measurements. AB - OBJECTIVE: To determine the interexaminer repeatability of the ulnar antidromic sensory nerve conduction velocity (NCV). DESIGN: Test-retest design. Based on a randomization list of various combinations and sequences from 2 of a total of 3 examiners, the measurement was repeated within half an hour by a second examiner blinded to the results of the first examiner. SETTING: Outpatient department. PARTICIPANTS: Twenty-four consecutive healthy subjects (mean age, 38 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The intraclass correlation coefficient (ICC) and the coefficient of repeatability (CR) were determined for the below elbow to wrist (BE-to-W), above elbow to below elbow (AE-to-BE), and axilla to above elbow (AX-to-AE) segments. RESULTS: The ICC was .42 for the BE-to W, .15 for the AE-to-BE, and -.05 for the AX-to-AE segment. The CR was 12.2m/s for the BE-to-W, 16.2m/s for the AE-to-BE, and 21.4m/s for the AX-to-AE segment. CONCLUSIONS: During the assessment of the antidromic sensory NCV of the ulnar nerve, a moderate amount of interexaminer variability must be taken into account for the BE-to-W segment. More proximally, an extremely large amount of interexaminer variability must be taken into account. This calls into question the usefulness of the antidromic ulnar sensory NCV for the AX-to-AE and AE-to-BE segments. PMID- 16213253 TI - A systematic review of the outcomes of cardiovascular exercise programs for people with Down syndrome. AB - OBJECTIVE: To determine if cardiovascular exercise programs are beneficial and safe for people with Down syndrome. DATA SOURCES: Electronic databases were searched from the earliest time available through to October 2004 using the following key words: Down syndrome or trisomy 21 in combination with physical fitness, exercise, physical activity, exercise therapy, exercise training, physical training, and aerobic. Additional articles were identified by manual searching and citation tracking. STUDY SELECTION: Two reviewers independently assessed the articles identified in the initial search for the following inclusion criteria: (1) participants with Down syndrome, (2) an exercise program that conformed with the American College of Sports Medicine guidelines for increasing cardiovascular fitness, (3) assessed changes in body structure or function, activity limitation, or societal participation, and (4) used a prospective clinical controlled research design with or without random allocation to groups. Trials of low methodologic quality were excluded (PEDro score, <4). Of the 156 articles initially identified, only 4 met the inclusion criteria and underwent detailed review. DATA EXTRACTION: Data relating to changes in body structure and function, activity limitation, participation restriction, and contextual factors from the included studies were independently extracted by the reviewers on a standardized form. Study quality was assessed using the PEDro scale. DATA SYNTHESIS: Meta-analyses found that cardiovascular exercise programs were effective in increasing peak oxygen consumption (d=.75; 95% confidence interval [CI], 0.34-1.15), peak minute ventilation (d=.71; 95% CI, 0.15-1.28), the maximum workload achieved (d=.96; 95% CI, 0.45-1.45), and the time to exhaustion (d=.72; 95% CI, 0.29-1.15) in people with Down syndrome. No changes were found for body weight (d=.09; 95% CI, -.39 to .57). No adverse effects were reported in any of the studies. CONCLUSIONS: The result of our review support the use of programs designed to improve cardiovascular fitness among people with Down syndrome. However, because only 4 studies were included, the findings need to be interpreted cautiously. High-quality randomized controlled trials should be completed in the future to determine the effect of these programs on activity and social participation. PMID- 16213254 TI - Cardiac rehabilitation after heart transplantation in a patient with Becker's muscular dystrophy: a case report. AB - Becker's muscular dystrophy (BMD) is associated with abnormal cardiac findings in 75% of cases; up to one third will develop ventricular dilatation leading to congestive heart failure, at times necessitating cardiac transplant. Candidates are selected from a base of heart failure patients who are usually New York Heart Association (NYHA) class III or IV. Treatment in a phase II cardiac rehabilitation program after transplantation is associated with functional improvement in patients without BMD, but there are no reports of patients with this disorder. We present the case of a 38-year-old man diagnosed with BMD with associated dilated cardiomyopathy. The patient was a NYHA class IIIa and underwent orthotopic cardiac transplantation for intractable heart failure followed by treatment in a phase II cardiac rehabilitation program. At the end of cardiac rehabilitation, his 12-minute walking distance had improved from 716.28 to 929.64 m (30% improvement), he had increased his conditioning metabolic equivalent level from 3.5 to 5.5 (55% improvement), he had a weight loss from 81.65 to 78.93 kg, and his body mass index changed from 23 to 22 kg/m2. The patient now has returned to work, is using a stationary bicycle once a day for 30 minutes, and is walking 1 hour a day. This suggests that treatment in a cardiac rehabilitation program is effective in patients with BMD after cardiac transplant. PMID- 16213255 TI - Saphenous nerve injury following medial knee joint injection: a case report. AB - Knee joint injection is a procedure commonly performed for pain management of osteoarthritis. Although several techniques have been described, it is usually performed by either medial or lateral approach with the lower limb extended on the examination table. We present the case of a patient who developed saphenous neuropathy following knee joint injection via medial approach. The clinical picture suggests that the needle pierced the nerve during the procedure. The patient was moderately obese. This is the first case report in the literature of saphenous nerve injury following medial knee joint injection. Practitioners should be aware of this complication in choosing an approach to the knee joint injection, especially when the patient is obese and the anatomic landmarks are obscured. PMID- 16213256 TI - A self-contained, mechanomyography-driven externally powered prosthesis. AB - The measurement of the low-frequency (5-50 Hz) "sounds" or vibrations produced by contracting muscles is termed mechanomyography (MMG). As a control signal for powered prostheses, MMG offers several advantages over conventional myoelectric control, including, nonspecific sensor placement, distal signal measurement, robustness to changing skin impedance, and reduced sensor costs. The objectives of this study were to demonstrate 2-function prosthesis control based on a triplet of distally recorded, normalized root mean square MMG signals and to identify necessary future research toward full clinical implementation of MMG signals in upper-limb externally powered prostheses. A novel self-contained MMG driven prosthesis for below-elbow amputees was designed, implemented, and preliminarily tested on 2 subjects. This prosthesis was composed of specialized software and hardware modules that emulate a 2-site electromyography sensing system. Although the use of MMG signals for prosthesis control has been shown previously, we report, for the first time, successful control within a self contained unit in unconstrained environments. Specifically, essential requirements for practical use, such as standardized sensor attachment, basic noise elimination, and miniaturization of the system, have been achieved. Both subjects were able to voluntarily open and close the prosthesis hand with no significant delays from intention to action (approximately 120 ms). Quantitative analyses revealed 88% and 71% control accuracy for subjects 1 and 2, respectively. PMID- 16213257 TI - The history and activities of transplantation in Turkey. AB - The cornerstone events of transplantation history in Turkey are summarized herein. In 1975, we performed the first living-related renal transplant in Turkey. This was followed in 1978, by the first deceased donor kidney transplantation, using an organ supplied by Eurotransplant. In 1979 the law on harvesting, storage, grafting, and transplantation of organs and tissues was enacted; later that year, the first local deceased donor kidney transplantation was performed by our team. In 1988, another groundbreaking event in Turkey was successfully achieved; the first cadaveric liver transplantation; and in 1990, the first pediatric living-related segmental liver transplantation in Turkey and in the region by our team. One month later, an adult-to-adult living-related liver transplantation was successfully performed. Until now, we have performed 1506 kidney and since 1988, 121 liver transplantations. During 29 years of solid organ transplantation history in Turkey, 6686 kidney transplants have been performed nationwide in 28 different centers; 696 livers; 13278 corneas; 2883 bone marrow; 132 hearts; and 15 pancreas transplants. In 2001, the health ministry established the National Coordination Center as an umbrella organization to promote transplantation activities, especially for deceased donor organ procurement. Transplantation activities are accelerating day by day all around the country, but deceased donors are still far below the desired rates. Improvements in the fields of education and coordination should increase the quality and the quantity of transplantation activities. PMID- 16213258 TI - Improving transplantation programs and patient care. AB - Organ transplantation is the preferred mode of replacement therapy. Currently acceptable 5-year posttransplantation survival rates are 85% for renal grafts, 70% for liver, and 65% for heart. Organ donation, however, remains a significant factor in organ transplantation, as humans are the only possible donors and the numbers of patients on waiting lists remain high. Several factors affect organ transplantation in countries in the Middle East Society for Organ Transplantation (MESOT) region, including inadequate preventive medicine, differing health infrastructures, poor awareness within the medical community and lay public regarding the importance of organ donation and transplantation, a high level of ethnicity, and poor government support of organ transplantation. Moreover, there is lack of team spirit among transplant physicians, a lack of coordination between groups that manage organ procurement and the transplant centers, and a lack of effective health insurance coverage for many persons. Three models of organ donation and transplantation exist in the MESOT region-the Saudi, Iranian, and Pakistani models. Living-organ donation, the most widely practiced form of donation in countries in the MESOT region, includes kidney and partial liver. Cadaveric organ donation has significant potential in the MESOT region. However, numerous obstacles must be overcome in MESOT countries. Resolution of these obstacles will require continuous work on many fronts. Experiences from all sources must be incorporated into new and improved models that can overcome current inadequacies. Solutions will require continued focus within the medical community, steady support from the lay public and religious institutions, as well as governmental assistance. PMID- 16213259 TI - Low tacrolimus dose requirements in renal transplant recipients in the omani population: implications for pharmacogenetics? AB - Clinical observations suggested that the dose requirements for some immunosuppressive drugs might be lower in some Eastern populations. We performed a study of 50 among 162 patients who were converted from a cyclosporine-based immunosuppression regimen to one based on tacrolimus. The reasons for conversion were as follows: acute rejection (n = 23), hirsutism and gingival hypertrophy (n = 12), chronic allograft nephropathy (n = 4), hypertension (n = 3), and other reasons (n = 8). Target levels were 10-15 ng/mL for patients with acute rejection and 5-8 ng/mL for patients after the first year of transplantation. We measured 180 tacrolimus blood levels in 44 patients using the IMx Abbott system of mean weight 64.5 kg and mean daily dose of 4.89 +/- 2.56 mg with mean blood levels of 11.31 +/- 5.22 ng/mL. Thus, to achieve the above-mentioned levels the average dose was 0.076 mg/kg, which is lower than that generally recommended to achieve 5 8 ng/mL levels, namely, 0.15 mg/kg. In this study, we did not address the reasons for these findings. The possibilities include dietary factors influencing the absorption of tacrolimus, and genetic polymorphisms in the Omani population that affect drug levels through several possible pharmacogenetic/genomic mechanisms. We intend to perform such studies in the future. PMID- 16213260 TI - Relationship between chronic inflammation and cardiovascular risk factors in children on maintenance hemodialysis. AB - Cardiovascular disease is one of the most important causes of morbidity and mortality in children with end-stage renal failure. Chronic inflammation and malnutrition have been suggested to be risk factors for cardiovascular disease. However, to date, biomarkers of inflammation have not been well studied in children. The aim of this study was to investigate the relation between chronic inflammation and cardiovascular risk factors in children on hemodialysis therapy. Twenty-seven patients on hemodialysis (14 girls, 13 boys) of mean age 15.3 +/- 2.4 years and 20 healthy children (13 girls, 7 boys) of mean age 14.3 +/- 2.7 years were included the study. C-reactive protein (CRP), albumin, prealbumin, transferrin, ferritin, and fibrinogen were measured as the markers of inflammation. The levels of CRP, ferritin, and erythrocyte sedimentation rate among hemodialysis patients were significantly higher than those of control subjects (P < .001 for all). Albumin and transferrin levels were found to be lower than those of control group (P = .02 and P < .001, respectively). CRP levels were negatively correlated with albumin, prealbumin, apoprotein A1, HDL, and hemoglobin levels, and positively correlated with erythropoietin/Htc ratios. This study suggests that hemodialyzed children are exposed to chronic inflammation. In addition, CRP may be an indicator of chronic inflammation related to cardiovascular risk factors, such as malnutrition, dyslipidemia, and anemia. In conclusion, we suggest that the risk of cardiovascular disease could be reduced by defining markers of chronic inflammation and malnutrition in hemodialyzed children and by taking necessary measures at an early stage. PMID- 16213261 TI - Effects of smoking and blood eosinophil count on the development of arteriovenous fistulae thrombosis in hemodialysis patients. AB - Arteriovenous fistulae (AVF) failure is the most common cause of morbidity and hospitalization in hemodialysis (HD) patients. The purpose of this study was to determine the effects of smoking and blood eosinophil count on the development of AVF thrombosis in HD patients. This cross-sectional study included 141 patients (M/F 80/61; age 43.4 +/- 11.6 years, HD duration 7.7 +/- 4.4 years). The following were analyzed as possible risk factors for AVF failure for all patients: demographic features, dialysis time, smoking, medications, body mass index, comorbid diseases, and various laboratory parameters (whole blood count and serum levels of albumin, calcium, phosphorus, uric acid, C-reactive protein, ferritin, and parathyroid hormone). AVF thrombosis was detected in 60 patients; in contrast, 81 patients had no thrombosis. Distributions of age, gender, and HD duration were similar between both groups. Univariate analysis showed that snuffbox AVF location (P < .0001), higher blood eosinophil count (P < .0001), smoking (P < .01), and higher hematocrit level (P < .05) were all associated with AVF thrombosis. According to multivariate analysis by logistic regression models, eosinophil count (RR = 1.005, P < .05) and snuffbox location (RR = 5.970, P < .05) were predictors of AVF thrombosis. When AVF location was excluded from the analysis, smoking (RR = 4.140, P < .01) and high blood eosinophil count (RR = 1.006, P < .005) were independent risk factors for thrombosis. Our study indicates that smoking and high blood eosinophil count may contribute to the development of AVF thrombosis. PMID- 16213262 TI - Vitamin D receptor BsmI and TagI gene polymorphisms in a Turkish ESRD population: influences on parathyroid hormone response. AB - BACKGROUND: Clinical presentation and complications of end-stage renal disease patients are influenced by many environmental and genetic factors. In this study we sought to define the frequencies of BsmI and TagI vitamin D receptor gene polymorphisms and their influences on clinical presentations in the Turkish end stage renal disease population. METHODOLOGY AND PATIENTS: Hemodialyzed patients (n = 186; 111 male, 75 female) were genotyped for the insertion/deletion BsmI (B -> b, restriction site, exon VIII --> IX), TagI (T --> t, 352 exon IX) vitamin D receptor gene polymorphisms. The previous 12 months of laboratory values (C reactive protein, intact parathyroid hormone, albumin, calcium, phosphorus, CaxP product) and clinical findings (vitamin D requirement, body weight) were analyzed retrospectively. RESULTS: Mean age and follow-up periods were 42.1 +/- 12.6 years and 76.3 +/- 43.9 months, respectively. Polymorphism percentages were BsmI; BB/Bb/bb: 28.9/65.3/5.8% and TagI; TT/Tt/tt: 36.7/60.5/2.8%, respectively. Further analysis revealed that the TT variant of TagI was related to hyperparathyroidism (P < .05). Analysis of the data after regrouping patients according to iPTH levels (0 to 249; 250 to 499; > or =500 pg/mL) and hemodialysis duration (<60 versus > or =60 months) revealed an influence of TT variation on hyperparathyroidism as a function of increased hemodialysis duration and higher iPTH levels (P < .005). CONCLUSION: TT variants of the TagI vitamin D receptor gene influence the development of hyperparathyroidism in hemodialysis patients, an influence that becomes more evident in patients with longer hemodialysis duration. PMID- 16213263 TI - Apolipoprotein a polymorphism predicts lipoprotein a concentration in renal transplant recipients. AB - Increased serum lipoprotein(a) is an independent risk factor for atherosclerosis in renal transplant recipients. Higher levels may be due to genetic factors, for example, apolipoprotein A isoforms and/or environmental states such as drugs and diets. We evaluated 75 renal transplant recipients including 30 men and 45 women of overall mean age of 30 +/- 7 years and transplantation duration of 57 +/- 10 months as well as 30 healthy controls for apolipoprotein A isoforms, lipoprotein(a) concentrations, serum triglycerides, serum cholesterol, serum creatinine, and serum homocysteine concentrations. High- and low-molecular-weight apolipoprotein A isoforms (>35 and <35 kringle 4) were observed in 71% and 29% of renal transplant recipients and 83% and 17% of controls. Average lipoprotein(a) concentration ratios between high- and low-molecular-weight apolipoprotein A isoenzymes were significantly greater in renal transplant recipients than in controls. Lipoprotein A and cholesterol concentrations that did not correlate with each other were not higher among the eight renal transplant recipients with creatinine levels greater than 1.8 mg/dL. Absolute levels in renal transplant recipients with failed grafts also were not different regarding the various apolipoprotein A phenotypes. Homocysteine levels were significantly higher with high-molecular-weight apolipoprotein A isoenzymes. A relationship existed between lipoprotein(a) and triglycerides, but not cholesterol: higher triglyceride levels were associated more with high-molecular-weight isoforms of apolipoprotein A (P = .027). Lipoprotein(a) concentrations are higher in low-molecular-weight isoforms of apolipoprotein but triglyceride levels and homocysteine concentrations are higher among the high-molecular-weight isoforms of apolipoprotein A. This finding could be used as a guideline to select the most appropriate drug for different apolipoprotein A isoforms. PMID- 16213264 TI - Angiotensin-converting enzyme and endothelial constitutive nitric oxide synthase polymorphisms in Turkish renal transplant population and possible influence on renal artery atherosclerosis and graft survival. AB - BACKGROUND: Renal transplant recipients are prone to accelerated atherosclerosis secondary to immunosuppressants, which may decrease graft survival. We sought to analyze the effects on renal graft survival of atherosclerotic degeneration in the renal artery and the influence of angiotensin-converting enzyme (ACE) endothelial constitutive nitric oxide synthase (ecNOS) gene polymorphisms. METHODS AND PATIENTS: Thirty three renal transplant recipients (25 men) of mean age 28.4 +/- 9.6 years, received organs from 11 living related donors and were followed for at least 36 months. Genotyping was performed for the insertion/deletion ACE (I/D), angiotensin (AGT) (M-->T, 235), angiotensine 1 receptor (A-->C, 1166), angiotensin 2-receptor (A-->G, 1223), and ecNOS (b-->a, intron4) gene polymorphisms. Renal artery biopsies were performed during transplantation surgery to analyze the presence of atherosclerosis. RESULTS: Pathological examination indicated that 18 donor specimens and nine recipient specimens had atherosclerotic degeneration. Survival analysis (36 months) indicated that graft survival rates of recipients who had atherosclerosis in the renal artery and who received an organ from donors with an atherosclerotic renal artery were shorter than in their counterparts (P = .02, P = .04, respectively). Comparison of genetic variations of recipients revealed that CC/TC variation of AGT was higher in patients with atherosclerosis (81% vs 53%, P = .03). There was no significant difference between groups in means of other gene polymorphisms. CONCLUSION: Renin-angiotensin system gene polymorphism analysis of patients in renal transplantation waiting list may provide information about allograft survival and posttransplant atherosclerotic degeneration at graft vasculature of young transplant recipients. PMID- 16213265 TI - Simvastatin therapy in lymphocyte cross-match-positive kidney transplantation candidates. AB - OBJECTIVES: Recent identification of several mechanisms by which statins decrease recruitment of monocytes and T cells into the arterial wall and inhibit both T cell and B-cell activation and proliferation in vitro prompted us to study the immunomodulatory effects of statins. In this study, we examined the effect of simvastatin therapy on lymphocyte cross-match positivity in kidney transplantation candidates. METHODS: Simvastatin therapy (20 mg/d) was administered to 25 patients (18 men, 7 women of mean age 34 +/- 11.7 years who displayed positive lymphocyte cross-matches between July 2002 and October 2004. The etiologies of end-stage renal disease were vesicoureteral reflux (n = 5), urinary stone disease (n = 4), glomerulonephritis (n = 6), amyloidosis secondary to familial Mediterranean fever (n = 1), and unknown (n = 9). RESULTS: The lymphocyte cross-match became negative in 10 patients 4-9 months, and successful kidney transplantation was performed in 6 of them. The serum creatinine levels of these patients ranged between 0.8 and 1.4 mg/dL. Two patients required higher doses, but none suffered from adverse effects. The remaining 4 patients are still undergoing pretransplantation evaluation. CONCLUSION: Simvastatin therapy seems to be a cost-effective and useful method for lymphocyte cross-match-positive kidney transplantation candidates compared with immunoadsorption or intravenous immunoglobulin use. PMID- 16213266 TI - Potential risk factors for hypersensitization reflected by panel-reactive antibodies in dialysis patients. AB - INTRODUCTION: The panel-reactive antibody (PRA) test has been considered to be a routine index of sensitization to human leukocyte antigens (HLA) in kidney transplant candidates. This study investigated the effect of potential risk factors and the time of blood sampling on PRA tests. METHODS: A total of 98 patients at two dialysis centers in Tehran were tested for PRA levels before and after dialysis sessions. We evaluated their history of potential sensitizing events and patient interviews for their association with PRA levels. Also we compared PRA levels obtained before and after dialysis. RESULTS: The mean age of the patients was 58.33 +/- 15.85 years. Only age and kidney transplantation history were correlated with PRA levels (r = .246, P = .014 and P = .0001, respectively). Logistic regression analysis revealed an association between age and PRA level (P = .037). Transplantation history was weakly correlated with PRA level (P = .076). History of pregnancy and transfusion, dialysis duration, gender, donor relation, and kidney allograft duration were not associated with PRA. PRA before dialysis sessions was significantly lower than that after dialysis (P = .0003). However, no difference was seen when divided into groups of negative/positive (PRA < 10% as negative) and high/low (PRA < 60% as low). CONCLUSION: Many factors expose patients to HLA as sensitizing factors. However, it seems that PRA level is not always predictable by such conditions. Furthermore, dialysis as a confounding procedure impacts PRA results; thus, when to obtain a blood sample is a crucial question. PMID- 16213268 TI - Intra- and postoperative complications of donor nephrectomies. AB - INTRODUCTION: Living-donor renal transplantation has become common worldwide. However, living-donor nephrectomy is not a complication-free procedure. This retrospective study reviews the intra-and postoperative risks and complications of donor nephrectomies at our institution between 1994 and 2002. METHOD: Two hundred fifty-seven consecutive donor nephrectomy patients were evaluated using medical records and anesthetic charts. RESULTS: The mean age of living donors was 42 +/- 12 years: 19 were older than 61 years, and 143 (55.6%) were women. Anesthesia for donor nephrectomy included general anesthesia, combined spinal epidural anesthesia (CSE), general + CSE, and general + epidural anesthesia. We observed 51 intra- and postoperative complications in 26 patients. The minor complication rate was 10.1%. The duration of surgery was 3.56 +/- 0.26 hours (range, 2 to 5 hours). Serum creatinine levels were increased significantly (P < .05) on postoperative days 1, 3, and 5 compared with the preoperative levels (P < .05). There was no mortality. CONCLUSIONS: Like other surgical operations, living donor nephrectomy is associated with intra- and postoperative complications. Although these complications are minor, maximal efforts must be applied in the anesthetic approach to minimize donor complications, and donors should be informed about potential risks. PMID- 16213267 TI - Concomitant nephrectomy of massively enlarged kidneys and renal transplantation in autosomal dominant polycystic kidney disease. AB - OBJECTIVES: We compared perioperative and intraoperative data of patients with end-stage renal disease (ESRD) due to autosomal dominant polycystic kidney disease (ADPKD) who received a renal allograft without native nephrectomy with ADPKD patients who underwent concomitant native nephrectomy of massively enlarged kidneys and renal transplantation to determine whether the latter approach is reasonable and safe. PATIENTS AND METHODS: From January 1987 to December 2003, 13 patients with ESRD due to ADPKD were stratified as 6 patients who underwent bilateral and 7 patients who underwent unilateral native nephrectomy in conjunction with renal transplantation (group A), versus 20 patients with ESRD due to ADPKD underwent renal transplantation without native nephrectomy (group B). Operative time, need for intraoperative transfusion, time to oral intake, duration of hospital stay, serum creatinine level on the day of discharge, readmission rate, and postoperative complications were compared for both groups. RESULTS: Mean intraoperative duration was significantly longer for patients in group A, but there was no statistically significant difference in the findings between both groups. CONCLUSIONS: Concomitant native nephrectomy of massively enlarged kidneys at the time of renal transplantation is reasonable and safe for patients with ESRD due to ADPKD. PMID- 16213269 TI - A controlled sequential evaluation of open donor nephrectomy versus classical and modified laparoscopic donor nephrectomy: an update. AB - This study sought to determine the procedure of choice for kidney retrieval for transplantation by comparing open donor nephrectomy to laparoscopic donor nephrectomy and modified laparoscopic donor nephrectomy and by analyzing intraoperative donor and recipient graft function parameters. In this single center, controlled, sequential analysis, 100 consecutive donor-recipient pairs were recruited, grouped according to surgical procedure, and operated upon between 1997 to 2004, as follows: group 1, open donor nephrectomy (n = 30), performed from 1997 to 2000; group 2, laparoscopic donor nephrectomy (n = 28), performed from 2000 to 2002; and group 3, modified laparoscopic donor nephrectomy (n = 42), performed from 2002 to 2004. Data were analyzed by type of operative procedure, graft function, length of hospital stay, and donor recovery time. Operative time was similar for all three surgical approaches. Warm ischemia times for open donor nephrectomy and modified laparoscopic donor nephrectomy were similar. Acute tubular necrosis occurred in 7% of patients in all groups. Donor recovery and lengths of hospital stay were significantly shorter for laparoscopic approaches. Donor complications were similar in numbers, differing only in complication type. Graft function and survival were similar for all three surgical approaches. We conclude that modified laparoscopic donor nephrectomy is the procedure of choice for living kidney retrieval. PMID- 16213270 TI - First living related kidney transplantation results in excellent outcomes for small children. AB - OBJECTIVE: The aim of this study was to evaluated the outcomes of living related kidney transplantation in small children. MATERIALS AND METHODS: Ten pediatric patients with body weights less than 10 kg received parental kidney transplants (five mothers and five fathers). An intra-abdominal approach was used in nine children and a retroperitoneal approach in one child. Bilateral, left, or right nephrectomy was performed in seven, two, and one child, respectively. Immunosuppression consisted of either cyclosporine (n = 7) or tacrolimus (n = 3) with either mizoribine (n = 4) or mycophenolate mofetil (MMF) (n = 5) or azathioprine (n = 1), and methylprednisolone (n = 10). Antilymphocyte globulin was used in the first series of four children; basiliximab in the most recent five children. RESULTS: All renal allografts functioned immediately after transplantation despite the mismatched size of the large renal allografts. Nine of 10 children were alive with a functional allograft at 6 to 196 months posttransplantation. One child died of intra-abdominal bleeding 5 days posttransplantation. One child has suffered chronic allograft nephropathy 11 years posttransplantation (serum creatinine 3.3 mg/dL). The remaining eight children display good renal function (serum creatinine = 0.2 to 1.43 mg/dL). Steroids were withdrawn in eight of nine children; one child continues on alternative-day therapy. One child (LD55) exceeded the mean standard height. The most recent height standard deviation (SD) scores were superior (-1.75 +/- 1.39 [ 3.83 to 0.54]; P < .0082) to those at transplantation (-2.91 +/- 0.79 [-2.00 to 4.14]). CONCLUSIONS: The outcomes of living related kidney transplantation in small children were excellent despite the operative risks and the difficulty of cardiovascular and fluid management. Transplantation for small children appears to result in much better quality of life and growth than dialysis. PMID- 16213271 TI - Long-term results of pediatric kidney transplantation at one center in Turkey. AB - Kidney transplantation is considered the treatment of choice for children with end-stage renal disease (ESRD). From November 1975 to June 2004, 80 of a total of 1477 kidney transplantations were performed in 78 pediatric patients. We retrospectively reviewed the records of 42 boys and 36 girls. Patient age ranged from 8 to 16 years (mean, 14.9 +/- 2.2). Sixty-three (78.7) grafts were from living donors and 17 (21.3%) from deceased donors. Two patients (2.6%) underwent preemptive transplantation, while 76 had preoperative renal replacement therapy with either hemodialysis in 62 (79.5%) or peritoneal dialysis in 14 (17.9%). Although the cause of ESRD could not be established in 30 cases, the other 48 showed the most common etiologies to be reflux nephropathy and glomerulonephritis. In conclusion, despite relatively poor socioeconomic conditions and health care problems in our country, the overall outcomes for pediatric patients at our transplantation center are good. We seek to perform more preemptive kidney transplantations in children with ESRD, and to increase our efforts to educate the Turkish public about organ transplantation and donation. PMID- 16213272 TI - Role of donor age and acute rejection episodes on long-term graft survival in cadaveric kidney transplantations. AB - Cadaveric donors can provide an effective solution to the problem of organ shortage, and many factors that may affect the functioning and survival of cadaveric kidneys have been studied. We aimed to clarify the impact of donor age and acute rejection episodes on long-term graft and patient survival in patients receiving cadaveric renal transplants. We retrospectively evaluated the long-term outcomes of 207 patients who had received cadaveric renal transplants between 1985 and 2004. Mean recipient age, HLA mismatch, mean donor age, delayed graft function (DGF), mean cold ischemia time, acute rejection episodes in the first 6 months after transplantation, and 1-, 3-, and 5-year graft survivals were evaluated. Two study groups were created according to donor age: group 1 (n = 126) was composed of patients receiving kidneys from donors younger than 50 years, and group 2 (n = 81) was composed of patients receiving kidneys from donors 50 years of age or older. Mean recipient age, HLA mismatch, and mean cold ischemia time between groups were not different. The DGF rate in group 1 was 40% (n = 50) and in group 2 was 46% (n = 37) (P > .05). The 1-, 3-, and 5-year survival rates of patients without acute rejection within the first 6 months after transplantation in group 1 (58/126; 46%) versus those in group 2 (46/81; 57%) were 95% versus 90%, 65% versus 60%, and 40% versus 35%, respectively (P > .05). The 1-, 3-, and 5-year graft survival rates of patients with acute rejection within the first 6 months in group 1 (n = 68) versus those in group 2 (n = 35) were 93% versus 89%, 71% versus 55%, and 44% versus 28%, respectively (P = .005). There was no significant difference in 1-, 3-, and 5-year survival rates between patients with DGF in both groups. Acute rejection episodes within the first 6 months after cadaveric transplantation, especially in patients receiving kidneys from donors older than 50 years, were shown to affect 5-year survival of the kidney graft. However, cadaver age alone had no negative effect on 5-year graft survival rates. Cadaveric donors older than 50 years may be a solution to the organ shortage in the treatment of end-stage renal disease. PMID- 16213273 TI - The effect of graft nephrectomy on long-term graft function and survival in kidney retransplantation. AB - We retrospectively evaluated the long-term results of 53 (3.5%) recipients who received second allograft among 1486 kidney transplants between November 3, 1975 and June 30, 2004. Two study groups were patients in Group 1 (n = 21) who underwent allograft nephrectomy and those in Group 2 (n = 32) who did not. We assessed demographic features, rejection rates throughout the follow-up period, and serum creatinine levels at 12 months as well as graft and patient survival rates, postoperative complications, time interval between transplantations, and HLA matches. Forty-three patients who underwent retransplantation received kidneys from living-related donors and the remaining 10 from cadaveric donors. Mean serum creatinine levels of Group 1 versus Group 2 were 1.8 mg/dL (range, 0.8 to 6.6 mg/dL) versus 2.1 +/- 1.1 mg/dL (range, 1.1 to 7.1 mg/dL). HLA-AB and HLA DR mismatches were 1.9 +/- 1.1 versus 1 +/- 0.6, respectively (P = .01). Acute rejection rates were not significantly different between Groups 1 (9/21, 43%) and 2 (12/32, 38%) (P < .05). The average intervals between the first and the second transplantations were 62 +/- 26 months in Group 1 (P = .02) and 32 +/- 11 months in Group 2. One-, 3-, and 5-year graft survival rates in Group 1 versus Group 2 were 83% versus 89% (P > .05); 64% versus 79% (P > .05), and 45% versus 68% (P = .04), respectively. In conclusion, we did not observe any advantage of graft nephrectomy before retransplantation. The length of the interval between the first and the second transplantations may have a negative correlation with second graft survival. PMID- 16213274 TI - Kidney retransplantation in comparison with first kidney transplantation. AB - INTRODUCTION: The aim of this study was to depict the outcome of second and third kidney allografts in comparison with first kidney allografts. METHODS: Among 2150 kidney transplantations are 103 second and 5 third transplantations. Demographic characteristics and survivals of retransplanted patients were compared with a randomly selected group of first kidney recipients, consisting of two cases matched with each retransplanted patient for age, gender, and date of transplantation. RESULTS: Retransplanted patients consisted of 78 men and 30 women of mean age 32.63 +/- 11.92 years. They had received kidneys from 91 living unrelated and 17 living-related donors. Median followup was 27 months. One-, 2-, 3-, and 5-year graft survivals were 81.4%, 78.9%, 78.9%, and 73.7% among retransplants, versus 92.9%, 91.5%, 89.8%, and 85.3% in the control group, respectively (P = .0037). Patient survival was 96%, 94.6%, 92.4%, and 87.8% in the retransplant group versus 93.1%, 92.4%, 90.9%, 87.4% in the control group, respectively (P = .63). Also, graft survivals were slightly lower in female compared to male retransplant patients (P = .09). No significant difference in survival rates was seen in different age groups. CONCLUSION: It seems that kidney retransplantation can yield desirable outcomes, albeit relatively lower graft survivals. PMID- 16213275 TI - Results in 158 consecutive cadaveric renal transplantations. AB - OBJECTIVES: We investigated the outcome of deceased donor kidney transplantations performed in a single center in a developing country. MATERIALS AND METHODS: A total of 158 patients (69 male and 89 female patients, including 32 children) received kidney grafts obtained from deceased donors between March 1996 and October 2004. Cadaveric renal grafts were transplanted after a cold ischemia time of 4 to 24 hours (mean, 12.5 hours). Retransplantation was performed in 19 recipients. Induction immunosuppression was achieved with antithymocyte globulin. The diagnosis of acute graft rejection was based on histopathological findings. RESULTS: Primary graft function was observed in 77% of cases. Posttransplantation complications were: surgical (n = 60; 38%), systemic bacterial and viral infections (n = 33; 21%), acute rejection (n = 47; 30%), and malignancy (n = 2; 1.3%). Seventeen recipients died with a functioning graft, and 23 more grafts were lost. The 7-year actuarial survival rates were 89% and 75% for recipients and grafts, respectively. CONCLUSIONS: The kidney transplantation program in Kuwait is steadily growing. Kidney grafts obtained from deceased donors contributed 28% of the transplantation activity and were associated with a high rate of primary function. Overall actuarial recipient and graft survival rates were comparable to those reported by larger centers. PMID- 16213276 TI - Effect of amyloidosis on long-term survival in kidney transplantation. AB - Amyloidosis is characterized by the accumulation of an amorphous material in various organs and tissues secondary to a variety of inflammatory, immune, infectious, and hereditary diseases. Since 1975, our transplantation team has performed 1470 renal transplantations. Between 1985 and July 2004, among 1159 kidney transplantations, 953 (82.3%) were from living donors and 206 (17.7%) from cadaveric donors. There were 32 recipients (28 men, 4 women; mean age, 31.4 +/- 1.7 years; range, 21 to 48 years) with amyloidosis, including, 28 (87.5%) who received grafts from living donors and 4 (12.5%) from cadaveric donors. Amyloidosis was secondary to familial Mediterranean fever in 22 (68.7%) patients and rheumatoid arthritis in 1 (3.1%). The remaining 9 (28.1%) patients had primary amyloidosis. The mean follow-up time was 51.2 +/- 5.7 months (range, 2 124 months). Mean HLA mismatch rate was 2.2 +/- 1. Twenty-six (81.2%) patients are alive at this time with functioning grafts, and a mean serum creatinine value of 2.1 +/- 1.5 ng/dL. The 1- and 5-year patient and graft survival rates were 90.6% and 84.3%, and 81.2% and 68.7%, respectively. We conclude that patients with amyloidosis may undergo kidney transplantation safely expecting outcomes similar to those patients who receive transplantations for other reasons. PMID- 16213277 TI - Lack of impact of human leukocyte antigen matching in living donor kidney transplantation: experience at Akdeniz University. AB - Lack of expansion of the deceased donor supply has resulted in a severe shortage of organs worldwide. Spousal donors are one possible alternative organ source for patients on the kidney transplant waiting list. Despite human lymphocyte antigen (HLA) matching between recipients and unrelated donors being poor, the reported survival rates for these grafts, including spouses, are comparable to those for grafts from living related donors and higher than those for deceased donor kidneys. In 2000, our renal transplantation program began accepting living donor recipient pairs with one or zero HLA matches. The purpose of this study was to assess this policy for accepting living unrelated donors. The 3-year graft survival rates for the transplants from living unrelated donors were similar to that for transplants from living related donors (log-rank = 0.078). The number of HLA mismatches did not significantly influence the survival rates for either of these groups of living donor transplants. Multivariate analysis revealed that dialysis duration (P = .057) and recipient age (P = .066) negatively influenced patient survival in living donor kidney transplantation. The graft and patient survival rates for the donor transplantations were higher than those for deceased donor transplantations. In light of these findings and considering the increasing problem of organ shortage, we conclude that living unrelated kidney transplantation should be performed, with strict guidelines. Spousal donation is the most favorable form of living unrelated renal transplantation. PMID- 16213278 TI - Renal allograft accumulation of technetium-99m sulfur colloid as a predictor of graft rejection. AB - Differentiation between rejection (the most common cause) and many other possibilities for detrimental effects on graft function represents a difficult issue to diagnose the cause of renal allograft dysfunction. This study was designed to determine whether technetium-99m sulfur colloid (TSC) accumulation predicted graft rejection. We prospectively studied 54 episodes of allograft dysfunction in 53 kidney transplant recipients who underwent TSC scintiscanning and graft biopsy. Visual analysis of TSC uptake compared uptake, in the allograft with that in the marrow of the fifth lumbar vertebra (L5). A 3+ result meant that allograft uptake was greater than L5 marrow uptake; 2+, the same; 1+, less and finally 0, no allograft uptake. Transplant accumulation of 2+ or more was considered consistent with rejection (P = .01). Allograft biopsies interpreted based on the Banff Working Classification showed rejection in 45 of 54 renal biopsies with 42 the biopsy-proven rejection episodes showing at least 2+ graft uptake. Furthermore, this nuclear medicine technique had a sensitivity of 93.3%, a specificity of 44.4%, a positive predictive value of 89.3%, a negative value of 57.1% and an efficiency of 83.3% for the diagnosis of renal allograft rejection. PMID- 16213279 TI - Doppler ultrasonography before and 6 to 12 months after kidney transplantation. AB - INTRODUCTION: Ultrasound examination of the kidney is relatively inexpensive and provides a way to assess renal location, contour, and size. Doppler ultrasonography is a noninvasive tool for screening renal artery stenosis. It not only provides kidney morphology data, but also describes hemodynamic changes associated with renal artery stenosis, such as increased peak systolic velocity and decreased resistance index (RI). The aim of this study was to compare the Doppler ultrasonographic changes between the donor's kidney before transplantation and the recipient's kidney at 6 to 12 months after transplantation. METHODS: We compared the results of Doppler ultrasonography in 20 kidney donors and recipients before and 6 to 12 months after transplantation. For this purpose the size, cortical thickness, echogenicity, anastomosis, mean pulsatility index (MPI), and RI of the kidney were recorded in potential donors before transplantation and in recipients at 6 to 12 months after transplantation for statistical analysis. RESULTS: There was more than a 10-mm increase in transplanted kidney length 6 to 12 months after transplantation in 75% of recipients. There was also more than a 10-mm increase in the width of the transplanted kidney in 80% of recipients. There was no significant change in cortical thickness between the donor and the recipient of the kidney. MPI and RI increased slightly after transplantation. There was more than 50% anastomotic stenosis in only 10% of transplanted kidneys. CONCLUSION: There was significant enlargement of the kidney size with a nonsignificant increase in MPI and RI of the transplanted kidney. Anastomotic stenosis was also less significant in our study. PMID- 16213280 TI - Value of pretransplantation cytokine profiles for predicting acute rejection in renal transplant recipients. AB - Episodes of acute rejection may represent an important risk factor for the development of chronic allograft nephropathy. Various studies have shown that pretransplant cytokine profiles in recipient blood are associated with transplant outcome. Serum samples were collected 24 hours before transplantation from 57 patients (38 men and 19 women of age 36 +/- 5 years) receiving kidneys from unrelated living donors. Additional samples were collected at 1 and 2 weeks after transplantation, as well as during every rejection episode. The immunosuppression consisted of a cyclosporine, prednisolone, and mycophenolate mofetil. Among the transplanted patients, 19 (33.3%) individuals experienced an acute rejection episode based on an increased level of serum creatinine and blood urea nitrogen during the first 14 days after transplantation. TGF-beta, IL-2 and IFN-gamma serum levels were determined by an ELISA method using Bindermed system kits. The mean concentration of TGF-beta before transplantation tended to be lower among patients with acute rejection episodes compared to those with stable graft (75,265 versus 85,394 pg/mL; P = .34) and at 1 week after transplantation (77,558 versus 84,390 pg/mL), although the differences were not significant. Among patients with rejection the mean IL-2 concentration was significantly higher before, at 1 week, and at 2 weeks after transplantation (15.0 versus 6.8 pg/mL, P = .005; 19.0 versus 4.9 pg/mL, P = .001; and 21.1 versus 4.7 pg/mL, P = .0001). The mean concentration of IFN-gamma was significantly higher pre- and at 1 and 2 weeks posttransplantation in patients with acute rejection episodes (161.1 versus 65.2, 175.6 versus 66.5 and 173.7 versus 77.1 pg/mL, all P < .001). In conclusion, evaluation of Th1 cytokines before transplantation may represent valuable predictive marker for an acute rejection episode. PMID- 16213281 TI - TH1/TH2 cytokine analysis in Iranian renal transplant recipients. AB - The pretransplant cytokine profile of donor and recipient blood and tissues may be associated with transplant outcome. A Th1 response is generally associated with transplant rejection, while a Th2 response may lead to tolerance and stable graft survival. A total of 56 (37 male and 19 female) patients of mean 36 +/- 5 years were candidates for living unrelated kidney transplantation. Serum samples were collected 24 hours pretransplantation as well as at 1 and 2 weeks posttransplantation. Immunosuppression consisted of cyclosporine, prednisolone, and mycophenolate mofetil. Among the transplanted patients, 19 (33.9%) individuals experienced an acute rejection episode, as proven by biopsy, as well as an increased serum creatinine and blood urea nitrogen, within 14 days after transplantation. We determined serum concentrations of interleukin (IL) 2 and interferon (IFN)-gamma for Th1 and IL4 and IL10 for Th2 by an enzyme-linked immunosorbent assay method (Bender med system kits, Germany). Among Th1 cytokines, the mean concentration levels for groups with versus without acute rejection were: IL-2 pretransplant 15 pg/mL vs 6.8 pg/mL, respectively (P = .005); IL-2 at 1 week, 19 pg/mL vs 4.85 pg/mL, respectively (P = .001); IL-2 at 2 weeks, 21.1 pg/mL vs 4.65 pg/mL, respectively (P = .0001); IFN-gamma pretransplant 161.1 pg/mL vs 65.2 pg/mL, respectively (P = .001); IFN-gamma at 1 week, 175.6 pg/mL vs 66.5 pg/mL, respectively (P = .001); and IFN-gamma at 2 weeks, 173.7 pg/mL vs 77.1 pg/mL (P = .001). IL-2 and IFN-gamma levels were significantly higher in the group with acute rejection versus those without acute rejection. In conclusion, these data suggest that cytokine analysis, especially of Th1 cytokines, might be a valuable prognotic index of kidney transplant outcome. PMID- 16213283 TI - Pretransplantation and posttransplantation body mass indices and prognosis in renal transplant recipients: low versus normal. AB - Body mass index (BMI) is strongly associated with outcomes in renal transplantation, independent of other risk factors. The aim of this study was to evaluate the impact of low BMI on graft survival in renal transplant recipients. The demographic and laboratory data as well as presence of acute or chronic rejection were retrospectively obtained for 115 recipients (80 men, 35 women) of mean age 34.56 +/- 11.14 years with posttransplantation follow-up duration of 5 years. Pretransplantation and one year posttransplantation BMIs were calculated. Patients were stratified to 2 groups according to their posttransplantation BMIs: group 1 had BMIs <19 kg/m2 (17.8 +/- 1.0; n = 23), and group 2 had BMIs > or =19 kg/m2 (23.7 +/- 1.8; n = 92). Twenty (87.0%) of 23 patients had low pretransplantation BMIs (P <.0001). Although mean serum creatinine levels at discharge after transplantation were lower among the low-BMI group (P <.03), the fifth-year levels were significantly higher in this group than in the normal-BMI group (P = .01). Follow-up serum albumin, triglyceride, and cholesterol levels were lower in group 1. According to the 5-year data, the percentages of recipients who suffered from chronic rejection (73.9% vs 20.7%; P < .001) and graft loss (73.9% vs 31.5%; P <.001) were significantly higher among group 1 than group 2. Multivariate backward analysis disclosed that BMI was closely associated with chronic rejection (P < 0.0001; odds ratio = 14.5; 95% confidence interval 4.3-49.6). In conclusion, a low BMI is an adverse prognostic factor after transplantation. To improve graft outcome, we recommend pretransplantation evaluation of recipient metabolic status, as well as early intensive dietary advice and follow-up for normalization of BMI. PMID- 16213284 TI - Effect of body mass index at time of transplantation and weight gain after transplantation on allograft function in kidney transplant recipients in Shiraz. AB - Chronic renal allograft dysfunction is the most common cause of graft loss, for which there are multiple risk factors, including obesity before transplantation, which is believed to lower long-term renal allograft survival. One hundred eighty two kidney transplant recipients were studied. Body mass index (BMI) at the date of transplantation was calculated. BMI values were classified into 4 categories: (1) patients with BMI <20, (2) BMI between 20 and <25, (3) BMI between 25 and <30, and (4) BMI > or =30. The minimum follow-up period in this study was 3 years after transplantation. The link between categorized BMI and the presence of renal allograft dysfunction and mortality within 3 years posttransplantation was investigated using independent sample t test. BMI at the date of transplantation showed statistically significant association with presence of renal allograft dysfunction and mortality within 3 years posttransplantation (P = .008, P = .01, respectively). BMI at the date of transplantation has a strong association with outcomes after renal transplantation. The extremes of very high and very low BMI are important risk factors for chronic renal allograft dysfunction; therefore, weight adjustment before kidney transplantation can be useful in improving the function of a transplanted kidney and increasing patient's survival. PMID- 16213282 TI - Switchability of neoral and equoral according to Food and Drug Administration rules and regulations. AB - According to the US Food and Drug Administration (FDA), if a drug product contains a drug substance that is chemically identical and is delivered to the site of action at the same rate and extent as another drug product, then it is equivalent and can be substituted (switchable) for that drug product. Methods used to define bioequivalence as stated by the FDA rules (FDA 21 CFR 320, 24) are (1) pharmacokinetic (PK) studies in healthy volunteers, (2) comparative clinical trials, and (3) pharmacodynamic (PD) studies (bioactivity). We evaluated the switchability of Equoral (IVAX-USA) with Neoral (Novartis Switzerland using all FDA rules. In a single oral dose, we undertook a comparative bioavailability study of Equoral (IVAX, USA) Neoral (Novartis, USA), and Neoral (Novartis UK). The pharmacokinetics of Equoral and Neoral were determined with blood levels at 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 16, 24, 30, 36, 42, and 48 hours. The area under curve (AUC), AUC extrapolated to infinity (AUC0-inf), rate of absorption (Tmax), extent of absorption (Cmax), half time (t1/2) of Equoral and Neoral were all within the 90% confidence interval of 80% to 125% boundaries. A comparative multinational multicenter clinical trial in stable renal transplant patients included 70 patients (22 women and 48 men) of mean age of 33 years (range, 26 to 43) was performed in Turkey, Lebanon, and Pakistan. In this study the ratios of LSM and the 90% confidence intervals for the Nontransformed/Parameters (AUC0-t, AUCinf, Tmax, and Cmax) of Equoral and Neoral SGC were 98% and 95%, respectively, which are within the 80% to 125% FDA acceptance range. For immunosuppressive drugs, the site of action is the lymphocyte and the measurable response is the decrease in lymphocyte count caused by the relative concentration of the drug in the lymphocyte. In a controlled switch, fixed-dose study, both Equoral and Neoral achieved the same concentration in the lymphocytes and caused the same degree of lymphocyte count reduction. The results of the testing (bioavailability-bioequivalence, clinical studies, and pharmacodynamic-bioactivity) required by FDA for interchangeability ("switchability") of immunosuppressive agents suggests that Neoral and Equoral are switchable. PMID- 16213285 TI - Improved mental health status in the first 2 weeks after kidney transplantation. AB - There is controversy whether mental health actually improves among kidney transplant recipients. The aim of this study was to investigate whether there was any significant change in mental status during the first 2 weeks after kidney transplantation. This cohort study involved 97 individuals who were transplantation candidates between 2003 and 2004. During the pretransplantation assessment, each individual completed the General Health Questionnaire (GHQ-28) and the Minnesota Multiphasic Personality Inventory, short form (MMPI). Posttransplantation subjects completed the GHQ-28 again. There were significant differences between mean total GHQ raw scores before and at 2 weeks after transplantation. Significant correlations were detected between a history of referral to a psychiatrist and a change in each of the 4 GHQ subscale scores. Our findings suggest that most kidney recipients show improved mental health in the early phase (first 14 days) after surgery. However, the results were not as positive as expected. Patients with a history of psychiatrist referral require special psychological attention. PMID- 16213286 TI - Impact of Ramadan fasting on renal allograft function. AB - Fasting during the holy month of Ramadan is a religious duty for all healthy adult Muslims. They are only allowed to eat and drink between sunset and dawn. This study was designed to find the effect of Ramadan fasting on allograft function. We prospectively studied 19 kidney transplant recipients who voluntarily chose to fast during Ramadan versus 20 matched recipients, who had not fasted for 3 consecutive years. Data were recorded before, during, and after the fasting month. The mean posttransplant periods in the fasting and control groups were 52.6 +/- 30.3 and 56.6 +/- 30.0 months, respectively. A statistical analysis showed no significant changes in serum creatinine concentrations before and after Ramadan 1.07 +/- 0.24 versus 1.08 +/- 0.22 mg/dL (P > .05) and 1.00 +/- 0.24 versus 1.03 +/- 0.28 mg/dL (P > .05) in fasting and control groups, respectively. The results did not show any adverse effects of fasting in recipients with stable renal function. In conclusion, our study suggests that fasting during the month of Ramadan is safe and has no significant harmful effects on kidney transplant recipients with normal renal function. PMID- 16213287 TI - Sirolimus as primary immunosuppression agent in kidney transplant recipients: Akdeniz University experience. AB - OBJECTIVES: Recently usage of sirolimus as the primary immunosuppressant is widening among kidney transplant recipients. We reviewed the clinical follow-up of patients transplanted at our center using sirolimus protocols. METHODS: Sirolimus including primary immunosuppressive treatment protocols were begun in February 2002. Among the 21 patients (15 men, six women) who received sirolimus, six patients were prescribed sirolimus + prednisolone; seven, sirolimus + mycophenolate mofetil + prednisolone; and eight, sirolimus + cyclosporine + prednisolone. The mean age of the patients was 32.9 +/- 7.3 years and the mean posttransplantation follow-up, 13.2 +/- 4.5 months. RESULTS: Three patients experienced acute rejection episodes, which were treated successfully with steroids. None of the patients had either hematologic or wound healing problems. Lymphoceles developed in eight patients. Serum creatinine level was 1.4 +/- 0.5 mg/dL at 12 months. There was a serious increase in serum cholesterol and triglyceride levels starting from the first month posttransplant (total cholesterol levels pretransplant and at 1 month, respectively: 159.3 +/- 29.5 and 255.7 +/- 52.3 mg/dL, P = .0001; triglycerides pretransplant and at 1 month, respectively: 146.9 +/- 89.5 and 215.1 +/- 102.5 mg/dL, P = .001). Despite routine antihyperlipemic treatment those high levels were maintained for 12 months. CONCLUSIONS: We achieved 100% graft and patient survival rates for 1 year among patients who were using sirolimus. But the most important role in defining the morbidity and mortality in this group of patients is cardiovascular events; for this reason the abnormalities in the lipid profile must be taken seriously. PMID- 16213288 TI - Tacrolimus plus low-dose mycophenolate mofetil in renal transplant recipients: better 2-year graft and patient survival than with a higher mycophenolate mofetil dose. AB - OBJECTIVES: Mycophenolate mofetil (MMF) has become more widely prescribed in recent years, but its adverse effects on the gastrointestinal system and bone marrow restrict its use in certain settings. The aim of this study was to compare the demographic features and clinical data for 173 renal transplant recipients who received tacrolimus (TAC) plus 1 g/d MMF (group I, n = 112) versus TAC plus 2 g/d MMF (group II, n = 61 patients) over a 2-year period. Each patient received similar TAC doses. METHODS: We compared demographic data and clinical data for each case: acute rejection (AR) episodes, chronic rejection (CR) episodes, death, graft loss, development of posttransplantation diabetes mellitus (PTDM), and posttransplantation hypertension rates. RESULTS: Demographic features were similar. There were also no significant differences between groups I and II with respect to number of AR episodes (17/112 vs 12/61, respectively), number of CR episodes (4/112 vs 1/61, respectively), PTDM, and hypertension rate (P > .05). Kaplan-Meier survival analysis revealed 2-year graft survival rates of 94% in group I versus 83% in group II. The corresponding 2-year patient survival rates were 100% in group I versus 91% in group II. The graft survival and patient survival rates in group I were significantly higher than those in group II (log rank 0.005 and 0.001, respectively). CONCLUSIONS: The 2-year graft and patient survival rates for the renal transplant recipients in this study suggest that the combination of a full TAC dose with 1 g/d MMF is a better choice than 2 g/d MMF. PMID- 16213289 TI - Mycophenolate mofetil in pediatric renal transplantation. AB - INTRODUCTION: Since kidney transplantation is the therapy of choice for children with end-stage renal disease (ESRD), we investigated the effects of mycophenolate mofetil (MMF) in pediatric renal transplantation. METHODS AND SUBJECTS: Two hundred sixteen children received renal transplants between 1985 and 2003: 100 patients received MMF with cyclosporine and prednisolone (cases), and 116 patients, azathioprine with cyclosporine and prednisolone (controls). RESULTS: The MMF group (100 patients) showed better graft survival and function than the AZA group (116 patients). Patients who received MMF immediately after transplantation experienced less graft loss and acute rejection episodes in the first 3 months after transplantation (P < .05). Patients who received MMF at the time of diagnosis of chronic rejection had stable renal function and remarkably better graft survival than those with chronic rejection who received AZA instead of MMF (P < .05). CONCLUSION: This study suggests that MMF may stop persistent graft dysfunction in chronic rejection, improving graft survival in the short and long terms posttransplantation. PMID- 16213290 TI - OKT3 treatment for steroid-resistant acute rejection in kidney transplantation. AB - Orthoclone (OKT3, Ortho Biotech Inc, USA) monoclonal antilymphocyte antibody is a powerful T-cell-specific immunosuppressive agent. OKT3 has been used for induction therapy in kidney and liver transplantation, as well as to treat acute or steroid-resistant acute rejection episodes (ARE). This study was a retrospective analysis of 43 renal transplant recipients who developed steroid resistant ARE and were treated with OKT3 between September 1994 and June 2004. The recipients were 36 men and 7 women of mean age 32.7 +/- 11.6 years (range, 19 to 48 years). The mean time from transplantation to OKT3 treatment was 7.2 +/- 6.7 months. Thirty-four episodes (79.1%) responded to OKT3 therapy with improved graft function, but the remaining 9 (20.9%) grafts did not respond. Among the 34 OKT3 responders, the mean serum creatinine decreased from 3.96 +/- 2.5 mg/dL to 2.45 +/- 1.77 mg/dL after treatment. Eleven (25.6%) of the 43 patients experienced minor side effects: fever, dyspnea, tachycardia, bradycardia. One patient (2.3%) developed acute pulmonary edema; one (2.3%), cytomegalovirus infection; and eight (18.6%), bacterial infections. The 1-, 3-, and 5-year graft survival rates for the 34 patients who responded to OKT3 therapy were 96%, 93%, and 85%, respectively. All patients are currently alive. The results indicate that OKT3 is a safe, effective treatment choice for steroid-resistant ARE in kidney transplantation. PMID- 16213291 TI - The area under the concentration-time curve versus trough and peak blood level monitoring in renal transplant recipients on cyclosporine. AB - OBJECTIVE: The area under the concentration-time curve of cyclosporine microemulsion is the best measure of the absorption and beneficial effects in renal transplant recipients. We sought to determine the best method of monitoring cyclosporine levels in these patients. METHODS: Prospective evaluation of peak cyclosporine blood levels and area under the curve monitoring were performed for 1 year in 65 renal transplant recipients (study group). Cyclosporine trough levels and peak cyclosporine blood levels were correlated with the calculated area under the curve. Cyclosporine trough levels were monitored in equal numbers of matched controls. RESULTS: There were no significant differences in the incidence of acute rejection, cyclosporine nephrotoxicity, proteinuria, serum creatinine levels, or graft and patient outcomes between the groups (P = .1). Peak cyclosporine blood levels guided by calculating the area under the curve were found to be 27% to 32% lower than previously reported. The correlation coefficient was <70% for cyclosporine trough levels (P < .02) and >90% for peak cyclosporine blood levels (P < .001) when related to the calculated area under the curve. The calculated area under the curve was approximately 6000 ng/mL/h following transplantation, gradually decreasing to approximately 3000 ng/mL/h at 1 year. Both appeared to the acceptable therapeutic values. CONCLUSION: Calculating the area under the curve using trough and peak blood levels versus using isolated readings for either of these levels alone is the most effective method of monitoring cyclosporine in recipients undergoing renal transplant. PMID- 16213292 TI - Single daily dose administration of cyclosporine in renal transplant recipients: a preliminary report. AB - INTRODUCTION: Cyclosporine microemulsion has been the mainstay immunosuppressive agent in renal transplantation for years. Since single daily dosing of cyclosporine is rarely used, the objective of this investigation was to evaluate the efficacy of a single daily dose versus twice daily dosing of cyclosporine in renal transplant recipients. METHODS: Retrospective evaluation of single-dose cyclosporine use was conducted for 15 renal transplant recipients for 12 months (study group). Equal numbers of matched renal transplant recipients were selected for age, sex, human leukocyte antigen mismatch, donor type, and immunosuppressive regimen (control group). Cyclosporine trough level and peak cyclosporine blood levels, 12-hour cyclosporine profile, and the area under the concentration-time curve were measured. RESULTS: There was a significant difference in cyclosporine peak blood level and calculated area under the curve after shifting to single dose cyclosporine (P = .001). In the study group, the mean area under the curve was significantly below the average therapeutic range before (3154 ng/mL/ho) versus 5532 ng/mL/h after shifting to the single-dose regimen (which was therapeutic). This value was 5749 ng/mL/h in the control group. Total daily cyclosporine dose was lower in the study group when compared with the control group at 6 and 12 months (P = .01). There were significantly fewer adverse effects in patients in the study group than in patients in the control group. CONCLUSION: We conclude that although cyclosporine dose should be individualized in renal transplant recipients, a single dose of cyclosporine has the added advantage of decreasing dosages and cyclosporine-related adverse effects while maintaining optimal graft function. PMID- 16213293 TI - Tacrolimus (FK506) versus cyclosporine microemulsion (neoral) as maintenance immunosuppression therapy in kidney transplant recipients. AB - The effects of the calcineurin inhibitors tacrolimus (FK506) and cyclosporine (Neoral) on graft survival, function, and metabolic profile were evaluated in 69 patients receiving Neoral (group 1) and 54 patients receiving FK506 (group 2) for maintenance immunosuppression following kidney transplantation. Recipient and donor demographics and induction therapy were comparable, except for a higher number of sensitized patients in group 2 (n = 13). Acute rejection timing, severity, and infection rates and types were similar in both groups. During hospitalization, at 6 months, and at 1 year following transplantation, no significant differences were noted between groups in fasting glucose, serum cholesterol levels, triglyceride levels, or need for insulin or antihypertensive therapy. Mean serum creatinine levels on discharge (1.42 mg/dL +/- 0.14 vs 1.68 mg/dL +/- 0.3), at 1 month (1.45 mg/dL +/- 0.1 vs 1.39 mg/dL +/- 0.11), 3 months (1.46 mg/dL +/- 0.09 vs 1.32 mg/dL +/- 0.14), and 1 year (1.29 mg/dL +/- 0.08 vs 1.19 mg/dL +/- 0.09), but not at 6 months (1.42 +/- 0.37 vs 1.10 +/- 0.07; P = .001), were comparable between groups. The 1-year patient and graft survival rates were 98.3% for group 1 and 94.5% for group 2. When evaluated for acute rejection, infection, and metabolic differences, we conclude that both tacrolimus and cyclosporine are effective and safe calcineurin inhibitors for short-term use in kidney transplantation. A similar study is proposed to evaluate the long-term effects of both agents. PMID- 16213294 TI - Early acute rejection episodes in renal transplantation in relation to immunosuppression protocols: an audit of 100 cases. AB - INTRODUCTION: Early acute rejection episodes (ARE) have deleterious effects on graft outcomes. The incidence of ARE in the first 3 months has been reported to be <20%. In a recent audit of ARE among 100 renal transplants, we observed the rates to be high (30%). We retrospectively collected details of donor type, induction therapy, immunosuppression medications, drug levels, HLA mismatches, acute tubular necrosis (ATN), and delayed graft function (DGF) to correlate with ARE and response to therapy. RESULTS: Thirty rejection episodes occurred after a mean period of 14.3 days after transplantation. Ninety-one patients had induction treatment with either antithymocyte globulin (ATG) or interleukin 2 receptor antibodies (IL2 Rab). The drugs included cyclosporine, mycophenolate, sirolimus, azathioprine, and prednisolone in these patients. There was no significant difference in ARE among the different drug protocols (30.7%-35.2%). Subjects with 4 or more HLA mismatches displayed more ARE (40.3%) compared with those with 3 or less (23%). Subjects with ATN or DGF immediately posttransplantation had a higher incidence of ARE (39.2%) than those without them (26.3%). Deceased donor recipients had a higher episode of ARE (45.1%) compared with live related donor recipients (25%). On stratifying the known risk factors for ARE, subjects with no risk factors had the least (22.2%) ARE compared with those with one (32.5%) or two (47.6%) risk factors. Subjects who failed to achieve adequate cyclosporine (C2) levels showed significantly higher rates of ARE (86.9%) than those with adequate or higher levels (8.6%). CONCLUSION: Higher HLA mismatches, DGF, deceased donor, and failure to achieve adequate cyclosporine levels were observed to be major risk factors for the development of ARE. PMID- 16213295 TI - Observations on quadruple immunosuppression maintenance therapy using rapamycin, low-dose cyclosporine, mycophenolate mofetil, and prednisone following ATG induction. AB - INTRODUCTION: We prospectively evaluated an immunosuppressive regimen consisting of rapamycin (Rapa), low-dose cyclosporine (CsA), low-dose mycophenolate mofetil (MMF), and prednisone (group 1) versus a regimen of CsA, MMF, and prednisone (group 2) in mismatched living related donor (LRD) and living unrelated donor (LUD) kidney transplantation. METHODS: Group 1 included 24 transplant recipients of eight mismatched LRD and 16 LUD, treated with Rapa, low-dose MMF, CsA, and prednisone. Group 2 included 53 transplant recipients (25 LRD, 27 LUD, and one cadaveric donor), treated with MMF, CsA, and prednisone. All patients in group 1 received a single bolus of rabbit-anti-human T-lymphocyte immune serum (ATG Fresenius 4 to 6 mg/kg). In group 2, patients received either a single ATG or an extended ATG course (3 to 5 days postoperatively). RESULTS: Acute rejection occurred in one patient in group 1 (4.2%) and in five patients (9.4%) in group 2, all of which resulted in graft loss. Serum creatinine was not significantly different between the two groups. CONCLUSION: The immunosuppressive protocol of Rapa, CsA, MMF, and prednisone with single-bolus induction ATG achieves excellent immunosuppression and graft survival with no apparent risks in the short and intermediate term. PMID- 16213296 TI - Invasive fungal infections in living unrelated renal transplantation. AB - In many developing countries, renal transplantation from paid, unrelated donors constitutes the main type of renal transplantation. Several medical and social problems are likely to occur in association with this practice. Among the many medical complications, invasive fungal infections are the most feared. In this report, we describe our experience with 3 patients who underwent living unrelated renal transplantation (LURTX) and developed this complication. One patient developed disseminated mucormycosis, 1 developed invasive aspergillosis (IA), and the third developed central nervous system (CNS) infection with Ramichloridium mackenziei. Two died within a few months after the diagnosis, whereas the third developed hemiplegia and is debilitated. PMID- 16213297 TI - Safety of caspofungin for treating invasive nasal sinus aspergillosis in a kidney transplant recipient. AB - INTRODUCTION: Invasive fungal sinusitis is a rare but often fatal infection in immunocompromised patients. Aggressive antifungal treatment is mandatory, but is not without risk. Caspofungin, an antifungal agent that is a member of the echinocandin family, an inhibitor of glucan synthesis in the fungal wall, is active against Aspergillus and Candidae infections. Although it works on the fungal wall, it does not affect mammalian cells; hence, its toxicity is minimal. CASE SUMMARY: This report describes a case of invasive Aspergillus sinusitis in a kidney transplant recipient with diabetes mellitus. The infection involved the apex of the right orbit causing optic nerve compression. The patient was treated with transnasal endoscopic decompression of the optic nerve and intravenous AmBisome (liposomal amphotericin B) for 2 weeks without clinical improvement. The combination of caspofungin and AmBisome administered for another 2 weeks yielded partial improvement. The AmBisome had to be discontinued due to deterioration of renal and hepatic function, but the patient completed a further 7-week course of caspofungin alone. Retro-orbital biopsy confirmed a complete response to treatment; the patient's renal and hepatic function returned to normal. CONCLUSION: This case indicates that caspofungin is effective to treat invasive Aspergillus sinusitis in kidney transplant recipients. This agent is well tolerated and safe with respect to renal and hepatic function. PMID- 16213298 TI - Preliminary report of a nationwide case-control study for identifying risk factors of tuberculosis following renal transplantation. AB - BACKGROUND: Tuberculosis (TB) is an important infection encountered posttransplantation, especially among patients in developing countries, where there are high incidences of morbidity and mortality. MATERIALS AND METHODS: One hundred and twenty subjects (1%) from 15 major kidney transplantation centers in Iran from 1984 to 2003 were compared with 440 controls who were matched for operative time, treatment center, and surgical team. RESULTS: Mean ages of research subjects and controls were 38.6 and 36.6 years (P = .04), respectively. The mean duration of pretransplantation hemodialysis was 29 months (range, 2 to 192 months) in research subjects and 20 months (range, 1 to 180 months) in controls (P = .003). Positive past history of tuberculosis was detected in 4 (3.3%) research subjects and in 7 (1.5%) controls (P = .2). Fifty-two research subjects (43.3%) and 241 controls (54.8%) had pretransplantation purified protein derivative of tuberculin less than 5 mm (P = .02). Mean dosages of initial and maintenance immunosuppressive drugs in research subjects and in controls were not significantly different. Sixty research subjects (50%) and 152 controls (34.5%) had rejection prior to diagnosis of TB (P = .03). CONCLUSION: To our knowledge, this is the first study that demonstrates an increased risk of posttransplant TB by prolonged duration of pretransplant hemodialysis and number of posttransplant rejection episodes. Further study is needed to clarify these findings specifically with respect to various immunosuppressive regimens. PMID- 16213299 TI - Outcome of hepatitis B and C virus infection on graft function after renal transplantation. AB - INTRODUCTION: Chronic liver disease resulting from hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is still a major concern in kidney recipients. It is unclear whether HCV antibody status and markers of HBV infection are associated with renal dysfunction. Thus, we designed a study to investigate the incidence of HBV and HCV infection after renal transplantation and whether these infections alter graft function. METHODS: Fifty-eight patients who underwent renal transplantation participated in the study. Serum creatinine and aminotransferase levels were measured with standard automated analyzers. Anti-HCV antibodies were detected with an enzyme immunoassay, and a reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to test for HCV-RNA. Serological markers for HBV (HBsAg and anti-HBc antibody) were detected by enzyme immunoassay. All samples from patients who were seropositive for HBsAg or anti HBc antibody were PCR-tested for HBV-DNA. A serum sample collected from living donors was tested for anti-HCV antibodies and serological markers for HBV. Serum creatinine and aminotransferase levels were also measured in living donors. RESULTS: Anti-HCV was not detected in serum samples of any cases before transplantation. However, 10 (17.2%) tested positive after transplantation. HCV RNA was detected in 2 of the 10 patients (3.4% of all patients). None of the pretransplantation serum samples tested positive for HBsAg. However, anti-HBc antibody was identified in 8 (13.8%) of the 58 patients.. No HBV DNA was detected in serum samples of the patients with anti-HBc or HBsAg-positive. HBsAg was only detected in 1 (1.7%) recipient after transplantation. None of the 58 patients showed clinical signs or symptoms of renal dysfunction during the study period. CONCLUSION: Our data suggest that, neither HBV nor HCV infection appears to cause or contribute to renal dysfunction in the early period (1 year) after renal transplantation. Nevertheless, a long-term consequence of chronic HBV or HCV liver disease or graft loss is not impossible in renal transplant recipients. PMID- 16213301 TI - Incidence of reactive antibodies against epstein-barr in a group of renal transplant patients. AB - Epstein-Barr virus (EBV) infection which is common among immunocompromised patients, may lead to life threatening lymphoproliferative diseases. In this study we examined the incidence and serologic status of EBV infection in 116 renal transplant patients including 84 males and 32 females as well as 72 normal volunteers. The time interval between transplantation and sampling was 1 month to 10 years. Twenty-two patients had a history of rejection. All cases were first transplants except for 3 second transplants. Four patients and no normals showed a positive PCR by a qualitative method. VCA IgM was positive in 11/116 patients (0.09%) and 3 of 72 (0.04%) normal volunteers. 99% (115/116) and 98% (65/72) of patients and normal controls were positive for VCA IgG. EA IgG was positive in 36/116 (31%) and 13/72(18%) of patients and normals, respectively. EBNA IgG was positive in 113/116 (97%) and 100% of patients versus normal controls, respectively. In all except one case with a positive VCA IgM there was a history of infectious mononucleosis-like syndrome. According to our previous data in more than 1000 renal transplant patients during more than 10 years, only one case of PTLD has been diagnosed (0.1%) which is lower than that reported. The high incidence of EBV seropositivity may contribute to this low incidence. The rate of EBV seropositivity in renal transplant patients was greater than in the normal population (P = .05). No association was observed between PCR and seropositivity and rejection or the type of treatment. After this study we began routine PCR and antibody testing in all renal transplant patients both pre- and posttransplant to determine the exact rate of reactivation versus primary infection which we plan to evaluate after 2 to 3 years. In conclusion we believe that the best easiest method to detect EBV infection in immunocompromised patients is VCA IgM ELISA; a qualitative PCR alone is not sufficient for this evaluation. PMID- 16213300 TI - BK virus nephropathy in renal transplant recipients in Kuwait: a preliminary report. AB - INTRODUCTION: BK virus nephropathy (BKVN) is a significant cause of graft loss among renal transplant recipients. The treatment outcomes of BKVN have been variably reported in the literature. PATIENTS AND METHODS: We prospectively investigated BKV infection and BKVN among a population of renal transplant recipients with suspected BKV infection. The 42 subjects who all had acute allograft dysfunction, were categorized in three groups: those with clinical, laboratory, and histological findings that did not suggest acute rejection, drug toxicity, or obstruction (group 1, n = 24); those with findings that suggested probable acute cellular rejection but did not respond to antirejection treatment (group 2, n = 10); and those whose renal histology suggested BKVN (group 3, n = 8). Polymerase chain reaction analysis was done to detect BKV DNA in urine and blood samples from each subject. BKV DNA was detected in 19 (45%) urine samples with 11 of these subjects (26.1% of total) having BK viremia as well. RESULTS: No evidence of BKVN was detected histologically in seven subjects with isolated BK viruria, while the others proved to be JC virus infections. Among the 11 subjects with BK viremia, eight had BKVN based on renal histology at the time of diagnosis with BKV infection, while the other three subsequently developed histological features of BKVN. BKVN developed after 5.3 +/- 2.5 (2 to 44) months after transplantation. The serum creatinine at time of BKVN diagnosis was 158.9 +/- 58 (87 to 285) micromol/L. All subjects were initially treated with a 50% reduction in immunosuppressive drug doses. Further decreases in immunosuppression were performed in all patients with close monitoring of renal function. All subjects were followed up for a of 18.2 +/- 5 (12 to 26) months. Two grafts were lost not due to BKVN, and one patient was lost to follow-up during this period. The latest serum creatinine in eight recipients is 113 + 20 (81 to 138) micromol/L, which is better than the renal function at diagnosis. CONCLUSION: The prevalence of BKVN in suspected BKV infection was 26%. Although the study period was short (30 months), BK viremia strongly correlated with BKVN, which seemed to be successfully treated with reduction in immunosuppression. PMID- 16213303 TI - Signs and symptoms of cytomegalovirus disease in kidney transplant recipients. AB - PURPOSE: To investigate the range of clinical presentations of cytomegalovirus (CMV) disease in kidney transplant recipients. MATERIALS AND METHODS: We retrospectively reviewed the records of hundred kidney recipients who developed CMV disease between 1984 and December 2002 for demographic characteristics, laboratory findings, and presenting signs and symptoms. RESULTS: The most common presentations were elevated serum creatinine in 74 patients, fever in 71, thrombocytopenia in 43, nausea in 32, vomiting in 25, elevated alkaline phosphatase in 24, leukocytosis in 22, and leukopenia in 21. Tissue involvement was relatively rare, but six patients had pneumonia, two had conjunctivitis, and one had vascular dermatitis. Four percent of the patients had received intravenous ganciclovir prophylaxis, and 7% had received oral ganciclovir prophylaxis. Fever was associated with number of hospitalizations (P = .006), elevated creatinine (P = .006), nausea (P = .017), vomiting (P = .031), and previous posttransplantation infections (P < .001). All the patients with conjunctivitis, pneumonia, pulmonary symptoms, and abnormal heart sounds and most of those with arthralgia, nausea, and vomiting were febrile during their CMV disease course. CONCLUSION: Our findings showed that leukocytosis should be considered as much as leukopenia when CMV disease is suspected. CMV-induced pneumonia is not common in renal transplant recipients compared to other organ transplant recipients. CMV invasion to other tissues is also rare. Finally, fever is a common symptom and important in assessing the severity and prognosis of the disease. PMID- 16213302 TI - A randomized prospective trial of oral versus intravenous ganciclovir for prophylaxis of cytomegalovirus infection and disease in high-risk kidney recipients. AB - PURPOSE: This study was designed to compare the efficacy and safety of oral versus intravenous ganciclovir in high-risk kidney recipients. METHODS: Thirty four, cytomegalovirus (CMV) seropositive recipients of kidneys from seropositive donors who had undergone antilymphocytic immunosuppressive therapy were assigned randomly to oral (1000 mg, three times a day, 12 weeks) versus intravenous (5 mg/kg, 2 weeks) ganciclovir prophylaxis. Follow-up was performed for 12 months. The patients were evaluated for clinical and laboratory outcomes regarding CMV serostatus, CMV disease, graft outcome, and ganciclovir side effects. RESULTS: Sixteen patients in the oral group and 14 in the intravenous group completed the study. CMV infection occurred in 6 (37.5%) and 5 (35.7%) cases in the oral and intravenous groups, respectively (P = NS). The mean interval between prophylaxis initiation and the first positive CMV Ag result was 3 +/- 2.19 months, with no significant difference between the two groups. Only two patients in the intravenous group experienced CMV diseases, which were not tissue-invasive. Acute rejection episodes were observed in nine out of 30 recipients, but it did not show any association with the prophylaxis regimen or CMV serostatus. The patients tolerated oral ganciclovir well; the compliance percent was 81.6%. No complication was reported. CONCLUSION: Oral and intravenous ganciclovir showed no significant difference to reduce the rate of CMV infection among high-risk kidney recipients. Oral ganciclovir was also effective and safe for the prevention of CMV disease. Moreover, it seems that CMV infection was not associated with acute rejection episodes. PMID- 16213304 TI - The role of early colonoscopy in CMV colitis of transplant recipients. AB - Cytomegalovirus (CMV)-associated diseases remain a major problem in transplant recipients. Early diagnosis is critical. Presentation of early CMV colitis can be mild and nonspecific in transplant recipients. Although serology is helpful in the diagnosis, sometimes it is inadequate. Because the endoscopic features of CMV colitis are specific, colonoscopy facilitates the histopathologic examination. We present the clinical properties and advantages of early colonoscopy in transplant recipients with CMV colitis. The study group included seven patients (six men, one woman of mean age, 36.7 years (range, 22 to 64 years) whose mean transplant duration was 12.3 months (range, 1 to 72 months). Six of the seven patients experienced an acute graft rejection treated with high doses of steroids; one patient had a herpes simplex virus infection. All patients were on steroid treatment with a various combinations of immunosuppressive agents, including cyclosporine, mycophenolate mofetil, and tacrolimus. All patients presented with mild diarrhea without any blood or mucous discharge. Four patients had fever exceeding 38 degrees C; two had abdominal pain. Stool examinations revealed normal findings in six patients, while one patient had white blood cells and amoebic cysts. Serum CMV IgM and CMV pp65 antigenemia were negative in five of seven patients and two had positive results. All patients showed typical colonoscopic and histopathologic findings compatible with CMV colitis. Standard ganciclovir treatment was successful in all patients. Early and rapid colonoscopy is beneficial for the early diagnosis and management of CMV colitis in transplant recipients. PMID- 16213305 TI - Outcome of Kaposi's sarcoma and graft following discontinuation of immunosuppressive drugs in renal transplant recipients. AB - PURPOSE: Owing to the use of immunosuppressive drugs, renal transplant recipients are at risk for malignancies including Kaposi's sarcoma (KS). Following the diagnosis, physicians tend to decrease the doses of immunosuppressive drugs to lower tumor progression rate. On the other hand, those who receive lower doses of immunosuppressive drugs are at a higher risk for acute rejection. In this study, we evaluated the outcome of KS on renal allografts following discontinuation or decrease in the doses of drugs. METHODS: Since 1984, 14 (nine men and five women) among 2000 cases of renal transplantation have been diagnosed as KS. In 11 patients, cyclosporine was completely discontinued, the dosage was decreased to half of the initial dose in other cases. Except one case, we discontinued either azathioprine or mycophenolate mofetil. RESULTS: During 57 months of follow-up on average, the serum creatinine level remained normal in 10 but increased in four cases. Kidney function deteriorated in two of these four patients at the beginning of study. Three patients died with normal serum creatinine levels. Discontinuation of immunosuppressive drugs caused complete remission of KS in all patients except one who received chemotherapy. CONCLUSION: Discontinuation of immunosuppressants following the diagnosis of KS caused complete remission of this cancer in almost all patients and seemed to be relatively safe for kidney graft function. PMID- 16213306 TI - Development of malignancy following living donor kidney transplantation. AB - BACKGROUND: Malignancy following renal transplantation is an important medical problem during the long-term follow-up. We studied some features of the cancers that developed in our patients. METHODS: We retrospectively reviewed all patients who underwent renal transplantation and developed malignancy from July 1984 to July 2004. RESULTS: The 2117 patients who underwent living donor kidney transplantation during the 19-year period had a mean follow-up of 81.1 +/- 61 months. During the follow-up, 38 patients (1.8%) developed cancer: 14 Kaposi's sarcomas, 11 lymphoproliferative diseases, four squamous cell carcinomas of the skin, two basal cell carcinomas, one breast, one ovary, one melanoma, one seminoma, one lung, and one ovary. Mean age at transplantation in the malignancy cases was higher than the other recipients (43.5 +/- 12.1 vs 32 +/- 13.9 years) (P = .000). A Kaposi's sarcoma occurred earlier compared with the other cancers (23 +/- 22 vs 62 +/- 44 months P < .05); most of these patients were over 40 years at transplantation (P < .05). We also observed that patients treated with mycophenolate mofetil developed cancer earlier than the others (19 vs 52 months; P = .001). None of the cases with lymphoma had a history of antilymphocytic agent therapy. The 10-year patient survival was 73%. CONCLUSION: The prevalence of cancer (1.8%) was among the lowest compared with other studies possibly due to implementing a living donor kidney transplantation program that required a low frequency of induction therapy. PMID- 16213307 TI - Malignancy in renal recipients. AB - BACKGROUND: Immunosuppressed organ transplant recipients are more susceptible to cancer than are persons in the general population. If malignancies of the skin are excluded for geographic variation, a cancer incidence of 4% to 7% in transplant recipients is usual. OBJECTIVES: We aimed to find the incidence, histopathological types, and outcome of malignancy in kidney transplant recipients in Kuwait. PATIENTS AND METHODS: Between 1972 and October 2004, more than 1500 kidney recipients were followed. After excluding recipients who left the country soon after transplantation, we reviewed the medical records of the remaining 1171 kidney recipients (724 male and 447 female patients of ages 3 to 76 years) at the time of transplantation. Kidney grafts were obtained from 968 living and 203 deceased donors. Records were retrospectively reviewed for the incidence, clinical presentation, histopathological patterns, and outcome of cancer. RESULTS: Fifty-six malignant lesions (4.8%) were diagnosed in 51 recipients (28 men and 23 women, aged 15 to 66 years), who had received grafts from 44 living and seven cadaveric donors. Malignancy was diagnosed 4 to 288 months after transplantation. The most commonest types were posttransplantation lymphoma and Kaposi's sarcoma. Posttransplantation cancer presented earlier in female and in adult recipients and following decreased donor transplantation. Kaposi's sarcoma appeared earlier than posttransplantation lymphoma or squamous cell carcinoma. Less than 40% of recipients with malignancy are alive. PMID- 16213309 TI - Urologic complications after renal transplantation. AB - BACKGROUND: Renal transplantation is associated with several nonimmunological problems. Although urologic complications may be serious and carry a high risk of graft loss, they are amenable to successful treatment if diagnosed early and treated properly. Their incidence in the literature varies from 2.5% to 15%. OBJECTIVE: We sought to assess the incidence, pattern, management options, and outcomes of urologic complications in 560 consecutive renal transplantations performed at a single center between November 1993 and October 2004. PATIENTS AND METHODS: Twenty-one (16 male and 5 female) recipients developed posttransplantation urinary complications at 2 days to 76 months after renal transplantation. Their kidney grafts were obtained from 13 living and eight deceased donors. Complications included ureteric stricture in 11 and urine leak in 10 recipients. Ultrasonography and isotope renal scanning were the main diagnostic tools. Complications were treated either conservatively, by percutaneous dilatation and stenting, or by surgical reconstruction. RESULTS: The incidence of urologic complications following renal transplantation in the present series was 3.7%. All cases were successfully treated with no graft loss secondary to these complications. CONCLUSIONS: Posttransplantation urologic complications are associated with a good prognosis if diagnosed early and properly treated. Percutaneous transluminal dilatation of ureteric stenosis in renal transplant patients has good initial success, low morbidity, few recurrences, and long-term effectiveness. PMID- 16213308 TI - Outcome of renal transplantation in children with low urinary tract abnormality. AB - INTRODUCTION: Patients with end-stage renal disease and lower urinary tract abnormality are often considered high risk for renal transplantation. METHODS AND SUBJECTS: To examine the degree of risk, we studied patients who received renal transplants between 1985 and 2003. Forty eight patients had congenital lower urinary tract anomalies and 168 patients comprised a control group without these anomalies. RESULTS: Mean age and distribution of sex were not significantly different between the case and the control group. Among patients with anomalies, 8% had delayed graft function; 75%, acute rejection; and 39.5%, chronic rejection. Among the controls 2.3% had delayed graft function; 59%, acute rejection; and 35%, chronic rejection. None of these differences was significant. Mean survival time was 6 years in affected patients and 7.3 years in the control group (P = .7). Among patients with anomalies the rate of graft survival in the first year after transplantation was 90%; and those in the third, fifth, and seventh years, 76%, 65%, and 40%, respectively. For the controls, the graft survivals were 88% at 1 year; 73% at 3 years; 70% at 5 years; and 49% at 7 years after transplantation. CONCLUSION: This study showed that a history of lower urinary tract anomalies had no effect on graft function. Graft survival was not different among these patients compared with patients free of these anomalies. PMID- 16213310 TI - Comparing Taguchi and anterior Lich-Gregoir ureterovesical reimplantation techniques for kidney transplantation. AB - Urologic complications are common in renal transplant surgery. Numerous innovations have been developed to circumvent ureterovesical anastomotic failure. In addition to the popular modified Lich-Gregoir technique, we evaluated Taguchi's method which is both quick and easy to perform. One hundred forty four patients were prospectively compared using Taguchi (n = 44) or the modified Lich Gregoir (n = 100) for anastomotic time, which differed significantly (10.2 minutes for Taguchi, vs. 24.6 minutes to Lich-Gregoir; P < .005). Minor complications, however, were less among Lich-Gregoir patients (P < .02). We concluded to continue using the modified Lich-Gregoir ureteroneocystostomy despite taking longer time to perform. PMID- 16213311 TI - Spontaneous kidney allograft rupture. AB - Spontaneous renal allograft rupture is one of the most dangerous complications of kidney transplantation, which can result in graft loss. This condition needs immediate surgical intervention. Conservative management has dismal results. Its prevalence varies from 0.3% to 3%. Rupture occurs in first few weeks after transplantation. Predisposing factors for graft rupture are acute rejection, acute tubular necrosis, and renal vein thrombosis. There are growing reports about successful results of repairing these ruptured kidneys. In this study, we reviewed the medical records of 1682 patients who received kidney allografts from living donors from 1986 through 2003. There were six (0.35%) cases of renal allograft rupture. All were preceded by acute graft rejection. They were treated with antirejection medications. In first three cases, the kidney allografts were removed because the procedure of choice in this situation is graft nephrectomy; but in three next cases we repaired the ruptured grafts with good results in two of them. In conclusion, the procedure of choice for kidney allograft rupture is graft repair. PMID- 16213312 TI - Male infertility after renal transplantation: achievement of pregnancy after intracytoplasmic sperm injection. AB - Chronic renal failure, dialysis, and immunosuppression after transplantation may cause reproductive failure. Although transplantation may reverse most sperm functions in the male, there is sufficient evidence in the literature that sperm motility may not be restored. Herein we present three cases with male factor infertility who underwent intracytoplasmic sperm injection (ICSI). There is a lack of evidence for use of ICSI for couples in whom the infertile husband had undergone renal transplantation. Case 1, a 25-year-old woman with a 7-year history of infertility had a 33-year-old husband, with asthenoteratospermia status-post renal transplantation 1 year prior. The wife delivered healthy twins at 36 weeks of gestation after three embryos were transferred. Case 2, a 34-year old woman with infertility of 6 years, had a 34-year-old husband transplanted 2 years prior and hospitalized for chronic rejection during ICSI treatment. He had severe oligoasthenoteratozoospermia. She is currently 20 weeks pregnant with a singleton after transfer of two embryos. Case 3, a 31-year-old woman with a previous spontaneous abortion and 3 years of failure to conceive, had a 41-year old husband status-post renal transplantation 16 years ago, currently in chronic renal failure treated with continuous ambulatory peritoneal dialysis. His sperm analysis showed oligoasthenoteratozoospermia. However, pregnancy did not occur after transfer of two embryos. Health status after renal transplantation influences sperm function. Because healthy sperm is required for fertilization and embryonic development, ICSI candidates after renal transplantation must be in optimum health. PMID- 16213313 TI - Unwanted pregnancy among kidney transplant recipients in Iran. AB - To investigate the incidence of unwanted pregnancy among kidney transplant recipients, we studied 86 pregnancies in 64 women with a transplanted kidney. Twenty-five pregnancies were unwanted (29.1%). Pregnancy was terminated by induced abortion in seven patients, and four pregnancies were lost due to spontaneous abortion with one intrauterine fetal death. Only 13 (52%) pregnancies resulted in a live birth. Most of the unwanted pregnancies occurred in women using coitus interruptus (92%) as the only method of contraception. It is concluded that because fertility greatly improves after kidney transplantation, it is necessary to have a family planning counseling session before surgery. If a patient is not interested in future pregnancy, an effective method of contraception should be offered. A woman who has decided against childbearing in the future may decide to have a tubal ligation at the time of transplantation surgery. PMID- 16213314 TI - Erectile function in end-stage renal disease before and after renal transplantation. AB - INTRODUCTION: Erectile function in end-stage renal disease (ESRD) and renal transplant patients is a challenging issue. In this study we evaluated the prevalence of erectile dysfunction (ED) according to standard questionnaires and paraclinical tests including Rigiscan. MATERIALS AND METHODS: We conducted a prospective, interventional, nonrandomized study of 15 consecutive male patients who underwent living donor renal transplants from March 2003 to June 2004. Before and after living donor transplantation we did hormone assays, blood ionogram and biochemistry, complete blood counts, u/a, international index of erection function 5 (IIEF-5), erection dysfunction intensive score (EDIS) tests as well as Rigiscan. RESULTS: The patient ages were between 21 and 50 (average 35.26) years, with an average length of ESRD of 4.31 years. Of the patients, 73.33% were smokers; 46.66% had ED; and 40%, hypertension. The most common blood groups were B-positive and O-positive (33.3% each). Mean testosterone and prolactin levels showed significant decreases after renal transplantation (P = .001 and P = .005, respectively). Mean blood glucose also decreased significantly (P = .035), despite previous reports that immunosuppressive drugs cause pseudodiabetes mellitus, mean cholesterol and triglyceride levels decreased after renal transplantation (P = .013, P = .0668, respectively). Urinalysis did not differ significantly after renal transplantation. Mean urea and creatinine levels were decreased significantly by renal transplantation (P = .000 and P = .003, respectively), but neither the mean values of uric acid nor the blood cell count were significantly different (P = .374). Mean hemoglobin and hematocrit levels were increased by renal transplantation, but it was not significant (P = .297 and P = .187, respectively). Mean potassium and phosphorus level were significantly decreased (P = .049 and P = .047, respectively), but mean sodium and calcium levels were not significantly altered (P = .773 and P = .536, respectively). Mean total and direct bilirubin and liver enzymes and alkaline phosphatase and LDH also did not change significantly. IIEF-5 was improved in 11 cases, unchanged in two cases, and worsened in another two cases. Nocturnal penile tumescence (Rigiscan test) was also improved in 11 cases, unchanged in three cases, and worsened in one case. The prevalence of erectile function was increased according to the EDIS question. CONCLUSION: Erectile function was improved after successful live donor renal transplant. PMID- 16213315 TI - Pregnancy in kidney transplant recipients. AB - PURPOSE: Our aim was to investigate kidney allograft, obstetric, and maternal outcomes in pregnant women undergoing kidney transplantation in our center. METHODS: Retrospective data on 74 pregnancies in 60 patients were reviewed and completed through phone interviews were compared with information on a control group of female kidney recipients. RESULTS: Mean age of patients at transplantation was 26.55 +/- 4.72 years and the median interval between transplantation and pregnancy was 27.5 months. Gestational period was 8 months. Live birth was the outcome in 43.2% of pregnancies; 9.5% led to still birth, 24.3% were aborted, and obstetrical data of the remaining were unavailable. Among the 11 patients who became pregnant within 12 months after transplantation, we observed seven live births and four abortions. None of pregnancies that were accompanied by acute rejection episodes (ARE) were successful. Twenty-six patients experienced at least one ARE versus 23 patients of the control group (P = NS). However, the first ARE occurred later in the pregnant group (P = .028). Chronic rejection and graft loss were seen in 24 and 18 study group cases and 17 and 17 control cases, respectively (P = NS). One-, 3-, 5-, and 10-year graft survivals were 100%, 96.5%, 94.5%, and 77.1% in the pregnant group versus 93.2%, 85.7%, 81%, and 64.7% in the control group, respectively (P = .07). CONCLUSION: Pregnancy in kidney recipients seems to be safe for kidney allograft recipients even within the first year posttransplant. Nonetheless, the outcomes of pregnancy in this group of patients is not always favorable, especially when rejection occurs simultaneously. PMID- 16213316 TI - High incidence of early posttransplantation diabetes mellitus in an eastern population. AB - The prevalence of diabetes mellitus (DM) in the Gulf region is among the highest in the world. In the general population of Oman, the rate is approximately 11.7% with an additional 6.1% of the population having an abnormal glucose tolerance. This study reviewed the data for 162 adults who received kidney transplants between 2001 and 2004. The immunosuppression regimen was cyclosporine, mycophenolate mofetil (MMF), and steroids. The mean age of the group was 40.2 years. Twenty-two patients (13.6%) had DM prior to transplantation. Within the first 2 months after transplantation, 45 (32%) of the remaining 140 patients required insulin, and 10 (7.1%) required oral agents. A further 16 patients (11.4%) displayed blood glucose levels >11 mmol/L, but required only a special diet for control. The data indicate that 50% of recent adult kidney recipients in Oman receiving cyclosporine develop posttransplantation DM. This major problem in our transplant population requires special attention. Protocols with minimal steroid use and/or steroid withdrawal may be beneficial for the Oman kidney recipient population. PMID- 16213317 TI - Incidence of posttransplantation diabetes mellitus in kidney transplantation: a single-center experience. AB - Posttransplantation diabetes mellitus (PTDM) is a well-recognized complication of renal transplantation. PTDM is reported to contribute to major infections and cardiovascular complications and leads to increased posttransplantation morbidity. The present study was conducted to evaluate the frequency of PTDM in our center, to identify the role of immunosuppressive therapy and other risk factors in the genesis of PTDM, and to assess the impact of PTDM on graft and patient survival. From December 1998 to December 2003 we followed-up 1200 renal transplant recipients, including 121 recipients with pretransplantation diabetes mellitus and 1079 recipients without diabetes. PTDM occurred in 203 patients (mean age, 35.4 +/- 5.9 years); 131 (64.5%) were male. Graft loss and mortality were significantly higher in patients with PTDM versus those without. The overall reported incidence of PTDM worldwide varies from 3.4% to 46%. The incidence in our center is 18.8%. We also found a significantly higher incidence of PTDM among patients receiving grafts from living-related donors. This may be related to the higher cumulative doses of immunosuppressive drugs administered (in part, due to the greater number of acute rejections) in these patients. PMID- 16213318 TI - Is HLA-DR6 a protective factor against posttransplantation diabetes mellitus? AB - BACKGROUND: Posttransplant diabetes mellitus (PTDM) has several pre- and posttransplant risk factors. METHODS: The incidence and risk factors of PTDM were retrospectively evaluated in 2117 kidney allograft recipients from June 1984 to March 2004. Type and dosage of immunosuppressive agents, pretransplant weight and human leukocyte antigen (HLA) phenotypes in PTDM patients were compared with 61 matched controls. RESULTS: Sixty-one cases (2.8%) developed PTDM requiring insulin or oral hypoglycemic therapy, out of which 47.5% were men and 52.5% were women, although only 35% of our overall recipients are women. Onset occurred at a mean of 489 days following transplantation. Patients receiving more than 15 mg/d prednisolone developed PTDM more often than those on less than 15 mg/d (P = .000). Similarly PTDM was more frequent among patients who received more than 300 mg/d cyclosporine compared with those on less than 300 mg/d (P = .015). Mean weight in PTDM cases and controls was 65 +/- 13.4 kg and 57 +/- 13.6 kg, respectively (P = .005). HLA-DR6 was observed in 12.2% of nonaffected subjects but in none of the PTDM group (P = .002). Conversely, HLA-DR8 was seen only in PTDM patients (P = .012). In addition HLA-A26 was more common among PTDM patients (P = .02) and HLA-DR52 more frequent in nonaffected subjects (P = .025). CONCLUSION: Our findings suggest that female sex, dosages of prednisolone and cyclosporine, pretransplant weight, and genetic factors are associated with an increased risk of PTDM. The rate of PTDM appeared to be independent of weight gain in the first year posttransplant. Protection against PTDM may be afforded by HLA-DR6 and possibly HLA-DR52. Conversely and higher incidence of diabetes has been associated with HLA-DR8 and HLA-A26. PMID- 16213319 TI - Erythrocytosis after renal transplantation: review of 101 cases. AB - Posttransplant erythrocytosis is increasingly recognized as a complication of kidney transplantation. In a retrospective analysis of 500 recipients, we observed 101 patients (20.2%) with persistent elevation of hematocrit value. It was more frequent in men (82.2%) than women (17.8%). It occurred 2 to 50 months after engraftment (mean value was 11.2 +/- 8.9 months), but most often developed in the first 24 months (86%). Spontaneous remission of established erythrocytosis was observed in all cases within 3 to 93 months. It frequently occurred in patients with a well-functioning renal graft; in 82.2% of cases the serum creatinine concentration was less than 1.5 mg/dL. It was 1.5 to 2 mg/dL in 15.8% of patients. There was no correlation between diabetes mellitus and erythrocytosis, compared with a control group. It was more common in patients who received cyclosporine compared to those who were not on cyclosporine. Predisposing factors included male gender, retention of native kidneys, cyclosporine use, and a rejection-free course with a well-functioning renal graft. In conclusion, posttransplantation erythrocytosis, a frequent problem in renal transplant patients, is a self-limited complication that can result in thromboembolic disease. PMID- 16213320 TI - Relationship of posttransplantation erythrocytosis to hypercalcemia in renal transplant recipients. AB - Experimental studies have demonstrated that calcium is an essential molecule in modulation of erythropoiesis. The aim of this study was to investigate the role of serum calcium levels on the development of posttransplantation erythrocytosis (PTE) among renal transplant recipients. We enrolled 155 patients (36 females/119 males; mean age, 34.9 +/- 9.7 years) with normal graft function who underwent renal transplantation between 1999 and 2002. All of the demographic features and various laboratory parameters were retrospectively analyzed as possible factors associated with erythrocytosis. PTE appeared in 43 (27.7%) patients during the follow-up period. Sixty-three (40.6%) patients developed hypercalcemia (corrected serum calcium level > or =10.2 mg/dL). Serum calcium levels tended to increase in patients with PTE, but significantly decreased in patients without PTE (10.6 +/- 0.6 vs 9.8 +/- 0.5 mg/dL; P < .0001). Similarly, hypercalcemia was more common among patients with PTE compared with patients without PTE (74.4% vs 27.7%; P < .0001). Hypercalcemic patients had a significantly higher frequency of PTE than normocalcemic patients (50.7% vs 11.9%; P < .0001). There were no differences in other laboratory and demographic data between the patients with and without PTE (P > .05). These findings suggest that hypercalcemia may lead to increased PTE in renal transplant recipients. PMID- 16213321 TI - Relationship between leptin and bone mineral density in renal transplant recipients. AB - INTRODUCTION: Leptin plays an important role in regulating appetite and energy expenditure and also functions in the neuroendocrine, hematopoietic, and immune systems, among others. Leptin may be involved in modulating bone mineralization. The relationship between leptin and bone mineral density (BMD) is not clear. This study examined the relationship between BMD and serum leptin levels in renal transplant recipients. MATERIALS AND METHODS: Forty-one patients (28 men and 13 women; age 16 to 55 years) were grouped according to percentile of serum leptin level hypoleptinemic (<5th percentile, n = 14), normoleptinemic (between the 5th and 95th percentiles, n = 19), or hyperleptinemic (>95th percentile, n = 8). The patients also were grouped according to lumbar z score) and total femur z scores (>-2 vs <-2 for both). RESULTS: The groups with different leptin statuses were compared with respect to age, sex distribution, and body mass index. Mean lumbar z score and mean lumbar BMD were higher in the hyperleptinemic group than in the normo- and hypoleptinemic groups (P < .05 for all). Considering the 42 patients overall, those with lumbar z scores >-2 had higher mean serum leptin/BMI than those with lumbar z scores <-2 (0.55 +/- 0.65 vs 0.18 +/- 0.23, respectively, P < .05). Serum leptin/BMI ratio was correlated with lumbar z score (r = .38, P < .05) and lumbar BMD (r = .32, P < .05). CONCLUSION: In conclusion, the data indicate that elevated leptin level is associated with increased bone mass at lumbar sites in renal transplant recipients. This suggest that increased leptin has a bone-sparing effect, especially in the lumbar region, in this patient group. PMID- 16213322 TI - Potential effects of 1,25-dihydroxyvitamin D3 in renal transplant recipients. AB - Besides its effects on bone metabolism, calcitriol has an important immunomodulatory effect, which may be protective for a renal allograft. Therefore, we evaluated the effects of oral calcitriol administration in renal transplant recipients. One hundred ten renal transplant recipients (78 men, 32 women) of mean age 35.2 +/- 11.4 years and mean posttransplantation follow-up of 50.7 +/- 22.9 months were entered into the study. Patients in group 1 (n = 57) received calcitriol therapy and patients in group 2 (n = 53) did not. The mean start of calcitriol therapy was 22.4 +/- 19.1 months posttransplantation. We restrospectively collected pretransplantation and posttransplantation laboratory and clinical data as well as creatinine levels before and after the initiation of calcitriol therapy at 6-month intervals for 2 successive years. There were no significant differences in terms of age, gender, immunosuppression, bone mineral densitometry, and follow-up. Our results showed that patients in group 1 had lower pretransplantation and postransplantation body mass index (P < .03; P < .03, respectively), lower posttransplantation third year parathyroid hormone levels (P < .02), and lower requirements for pulse steroid doses (P < .04). Using Friedman repeated measures variance test to analyze the effect of calcitriol, the increase in creatinine levels was significantly lower in group 1 (P < .04). There was no significant difference between follow-up time and calcitriol dose (P > .05). In conclusion, calcitriol therapy may reduce the rate of loss of renal function among patients receiving renal transplants. PMID- 16213323 TI - Assessment of bone structure in renal transplant recipients: comparison of phalangeal qualitative ultrasound and dual x-ray absorptiometry. AB - According to the WHO criteria many renal transplant patients display osteopenia or osteoporosis. Dual-energy X-ray absorptiometry (DXA), the standard method to assess bone mineral density (BMD), is not always available. Quantitative ultrasound (QUS) of the phalanx is an inexpensive, mobile, and radiation-free diagnostic alternative. Few data address the correlation of this method with DXA in renal transplant patients. This study assessed the value of QUS compared with DXA to detect changes in bone structure among renal transplant recipients. This cross-sectional study of 42 patients (22 women), of mean age 40.2 +/- 11.9 years, mean time since transplantation of 2.8 +/- 2.9 years, and mean dialysis time of 8.55 +/- 10.26 months, included. DXA for bone mineral densitometry of the hip (neck and total femur) and spine as well as QUS to measure the amplitude dependent speed of sound (Ad-SOS) in the phalanx. Using DXA, osteoporosis was observed in 19% of all patients: 9.5% in femoral neck, 9.5% in total region of the femur, and 9.5% in the spinal region. The sensitivity of Ad-SOS for osteoporosis diagnosis in the above regions were 100%, 75%, and 25%, respectively; its specificity was 45%, 43%, and 37%, respectively. There was no significant relation between the two methods for diagnosis of osteoporosis in any region. QUS of phalanx can be recommended for osteoporosis screening in renal transplant patients. Those suspected of osteoporosis should be examined by additional DXA measurements in order to establish the diagnosis. PMID- 16213324 TI - Risk factors for osteoporosis in young renal transplant recipients. AB - Osteoporosis is a frequent complication after renal transplantation. Although several risk factors have been defined in its pathogenesis, the parameters influencing this complication in young patients with functioning grafts have not been well defined. We sought to determine the possible risk factors for osteoporosis among 55 male and 27 female renal transplant patients of mean age 35.5 +/- 11.7 years with 68.8% recipients of living-related grafts. Bone mineral densitometry was performed in all patients at 1 year after transplantation. The study utilized the World Health Organization recommendations that define normal (group 1, n = 19), osteopenic (group 2, n = 24), or osteoporotic (group 3, n = 39) patients according to T-score values. Pre- and posttransplant data included gender, age at dialysis onset; age at transplantation; pretransplant dialysis duration; body mass index (BMI); serum calcium, albumin, phosphorus, parathyroid hormone (PTH), and C-reactive protein levels; lipid profile, cumulative doses of immunosuppressive drugs, and pulse steroid dose. Differences between groups 1 and 3 showed that patients with osteoporosis were younger (P < .003) and had started dialysis and underwent transplantation at a younger age than those without osteoporosis (P < .01, P < .003). In addition, pretransplant body weight (P < .02), posttransplant BMI (P < .01), and pretransplant PTH (P < .04), posttransplant total cholesterol, and high-density lipoprotein cholesterol levels were lower among group 3 (P < .004, P < .003). Young adults who started dialysis and underwent transplantation at a younger age were prone to osteoporosis. Additionally, high BMI and cholesterol levels seemed to be preventative for bone loss after transplantation. PMID- 16213325 TI - Influence of cyclosporine and tacrolimus on serum uric acid levels in stable kidney transplant recipients. AB - Although hyperuricemia is a well-known adverse effect of cyclosporine (CsA) treatment, there are contradictory data regarding the effect of tacrolimus on uric acid levels. The aim of this study was to examine the influences of CsA and tacrolimus-based treatment regimens on serum uric acid levels in 155 renal transplant recipients with normal allograft function who underwent renal transplantation between 1999 and 2002. Serum uric acid levels were recorded at 1, 6, 12, 18, and 24 months follow-up. The patients were treated with CsA-based (n = 73), tacrolimus-based (n = 47), or conversion from CsA-based to tacrolimus-based (n = 35) immunosuppressive regimens. Serum uric acid levels for patients in the CsA and tacrolimus groups were 6.3 +/- 1.6 versus 7.9 +/- 1.9 mg/dL and 6.5 +/- 1.8 versus 8.0 +/- 1.8 mg/dL at the study outset and 24 months, respectively. Both of the treatment regimens showed progressively increasing serum uric acid levels (P < .001). Serum uric acid levels of patients with treatment conversion from CsA to tacrolimus were 8.6 +/- 2.8 mg/dL before conversion and 8.1 +/- 1.9 mg/dL after conversion. There was no alteration in serum uric acid levels after the change of treatment (P > .05). These findings indicate that, compared with CsA, tacrolimus offers no advantage for serum uric acid levels in renal transplant recipients. PMID- 16213326 TI - Findings of Doppler sonography do not correlate with serum lipoprotein and total homocysteine concentrations in renal transplant recipients. AB - Atherosclerosis may be evaluated by structural or functional changes of the main arteries. We sought to investigate the probable associations of static and dynamic arterial changes with lipoprotein (a) and homocysteine levels, the two risk factors for atherosclerosis. Intima-media thickening and vasodilatory responses to nitroglycerine of the common carotid artery and the renal transplant artery were studied by color Doppler sonography in 75 renal transplant recipients and 30 controls. At 3, 5, and 10 minutes after 0.4 mg of sublingual nitroglycerine are measured resistive index and peak systolic velocity of the common carotid artery and renal transplant artery. Intima-media thickening in renal transplant recipients and controls were 0.86 +/- 0.34 mm and 0.74 +/- 0.14 mm (P = .05), respectively. Although intima-media thickness did not correlate with the duration of renal transplantation, it was significantly higher in older renal transplant recipients. Peak systolic velocity of common carotid artery was significantly decreased by nitroglycerine in the controls (81.8 +/- 16.7 m/s to 73.2 +/- 12.8 m/s, P = .03). This decrement was more obvious in renal transplant recipients, especially at 10 minutes (69.6 +/- 18.5 m/s vs 59.3 +/- 2 m/s, P = .01). These reductions did not correlate with intima-media thickening, latter of which also did not correlate with homocysteine concentrations, which were higher among renal transplant patients with creatinine more than 1.8 mg/dL. Basal resistive indices of the common carotid artery and renal transplant artery were higher among graft recipients with dysfunction than recipients with good function, (0.7 vs 0.59, P = .003). In conclusion, neither homocysteine nor lipoprotein(a) concentrations predict static and dynamic vascular properties. PMID- 16213327 TI - The significance of Tc-99m DTPA renal scintigraphy for the demonstration of progressive functional changes that accompany chronic allograft nephropathy. AB - OBJECTIVES: Chronic allograft nephropathy (CAN) is usually progressive; its natural course can only be modified in the initial stages. In this study, we graded Tc-99m diethylenetriaminepentaacetic acid (DTPA) renogram curves with respect to the perfusion/uptake pattern and correlated these findings with biopsy results in patients with CAN. METHODS: This study included 63 renal allograft recipients with biopsy-proven CAN. The agent used for renal scintigraphy was Tc 99m DTPA. Quantitative evaluation of perfusion included calculation of the ratio of peak perfusion counts divided by plateau counts (P:PL). Deterioration of renal function was accompanied with a gradual loss of a peak and plateau pattern. For the evaluation of uptake in relation to perfusion pattern, we graded the renogram curves into four based on the presence of a peak and plateau pattern and the presence of an uptake peak. RESULTS: In patients with CAN, the mean P:PL was significantly lower than that of the control group. The serial changes in successive grades of CAN in respect to uptake-perfusion pattern was a gradual loss of peak and plateau pattern followed by a decline in uptake. In recipients with high-grade CAN, an uptake peak was absent. CONCLUSIONS: Evaluation of Tc-99m DTPA time-activity curves revealed a progressive change in perfusion-uptake pattern in patients with CAN. According to our results, deterioration of perfusion preceded the decline in uptake. Serial renogram changes are thought to reflect initial hypoperfusion followed by increased intraglomerular pressure and finally glomerulosclerosis. These findings have implications for the pathophysiology and management of CAN. PMID- 16213328 TI - Living-related liver transplantation in pediatric patients. AB - INTRODUCTION: Many developments in surgical technique, immunosuppression, and patient selection criteria have led to improved long-term patient and graft survival in pediatric patients receiving liver transplants. In this study, we examined the early results of 26 pediatric recipients who underwent 26 liver transplantations between January 2003 and December 2004 at our institution. MATERIALS AND METHODS: The most common indications for liver transplantation were cholestasis in 10 patients (38.5%) and Wilson's disease in 8 (30.8%). Other indications were fulminant hepatic failure (4 patients, 15.4%), tyrosinemia (2 patients, 7.7%), Caroli disease (1 patient, 3.8%), and cryptogenic cirrhosis (1 patient, 3.8%). One recipient with Byler disease and two with tyrosinemia also had incidental hepatocellular carcinoma. RESULTS: Of 26 patients, 24 (92.3%) underwent living-related liver transplantation and 2 (7.7%) underwent cadaveric transplantation. The medical records of all patients were retrospectively reviewed. Twenty-two of 26 survived with excellent graft function, showing 91.2%, 86.4%, and 81.6% at 3, 12, and 24 months graft and patient survival rates, respectively. Sixteen patients (61.5%) developed various morbidities with biliary and vascular complications being the most common. Four patients (15.3%) developed bile leaks. Four patients (15.3%) developed hepatic artery thromboses. Five patients (19.2%) developed life-threatening infections. Four patients (15.4%) died during the study period, three owing to infectious complications. The other patient died due to acute respiratory distress syndrome. CONCLUSION: Despite technical difficulties and a donor organ shortage, the results of liver transplantation in pediatric patients with end-stage liver disease have demonstrated promising results at our institution. PMID- 16213329 TI - Duct-to-duct biliary anastomosis with a "corner-saving suture" technique in living-related liver transplantation. AB - OBJECTIVES: Biliary complications after living-related liver transplantation (LRLT) remain a major source of morbidity for recipients. We describe our technique and early results with 32 recipients who underwent LRLT with duct-to duct anastomoses during the last 2 years. METHODS: Between January 2003 and December 2004, 50 patients underwent liver transplantation in our center with overall patient and graft survival rate of 86.4% and 86.4%. Of 50 patients, 41 (82.0%; 17 adult and 24 pediatric) underwent LRLT, 32 (78.0%) of whom had duct-to duct biliary anastomoses with a "corner-saving suture" technique. RESULTS: Of 32 patients in whom duct-to-duct biliary anastomoses were performed, 4 (12.5%) had an anastomotic leak with 2 eventually developing bile duct strictures within 3 months. One patient required reoperation for a bile leak. All other anastomotic leaks and strictures were treated with percutaneous drainage and balloon dilatation with excellent outcomes. There was no long-term morbidity and no graft loss owing to biliary complications. Seven patients died during follow-up (0.5 to 25 months); 43 are doing well with optimal liver function in the early posttransplantation period. CONCLUSION: According to our early results, we recommend duct-to-duct anastomosis in LRLT when calibration of the ducts show suitable results and when there is no tension on the anastomosis site. Otherwise, Roux-en-Y hepaticojejunostomy should be performed to decrease risk of biliary complications. PMID- 16213330 TI - Living related liver transplantation for hepatocellular carcinoma in Egypt. AB - BACKGROUND: Living related liver transplantation (LRLT) for hepatocellular carcinoma (HCC) in cirrhotic patients has emerged as a rewarding therapy for a cure. Extensions of the Milan criteria have been proposed with encouraging results. PATIENTS AND METHODS: From October 2001 to June 2004, 47 adult patients with end-stage liver disease (ESLD) have been treated using LRLT, including 11 (9 males and 2 females) with HCC superimposed on hepatitis C virus (HCV)-related (n = 10) or hepatitis B virus-related (n = 1) cirrhosis. Their mean age was 50 years (range, 40-61). HCC was confirmed preoperatively in 9 subjects whereas it was an incidental finding in 2 cases. Alpha fetoprotein (AFP) levels were elevated in 5 of them. Radiologically, tumor number and sizes ranged from 1 to 2 nodules and from 1.5 to 7 cm, respectively. Five of the 11 subjects underwent pretransplantation tumor control therapy. RESULTS: Nine patients are alive, all of them being disease free during follow-up periods ranging from 6 to 30 months. Two subjects died: one of HCC recurrence at 1 year posttransplantation, and another of a pulmonary embolism on day 7. AFP levels decreased to normal values in 4 cases. Excluding the 2 incidental tumors, pathological examination of the explants revealed a higher number and larger size of the nodules in 3 and 5 cases, respectively. Microvascular invasion was documented in 3 explants, 1 of which experienced HCC recurrence and the other 2 received 6 cycles of Doxorubicin following normalization of their liver profile. Postoperative complications included the following: recurrent HCC (n = 1), recurrent HCV (n = 2), acute cellular rejection (n = 3), anastomotic biliary stricture (n = 1), and subphrenic collection (n = 1). CONCLUSION: Our current data confirm the efficacy of LRLT for treatment of HCC superimposed on liver cirrhosis. PMID- 16213331 TI - Twenty cases of adult-to-adult living-related liver transplantation: single center experience in Saudi Arabia. AB - BACKGROUND: The deceased donor organ shortage has forced surgeons to implement innovations, including living-related liver transplantation (LRLT). OBJECTIVE: To present the first 20 cases of adult LRLT in a single center in Saudi Arabia. METHODS: From November 2000 to May 2004, we performed 20 cases of LRLT. Eighteen donors were men and 2 were women. Their median age was 27 years. Seventeen of the recipients were men and 3 were women of median age 55 years. One patient received combined liver and kidney grafts. RESULTS: All cases had liver cirrhosis. Seven had hepatitis C; six, hepatitis B and C; three, hepatitis B; one, alcoholic cirrhosis; one, Bylar disease, one hepatic schistosomiasis, and one cryptogenic cirrhosis. Three cases had associated hepatocellular carcinomas. There was no donor mortality. In the recipients, the overall patient and graft survival was 85%. While 10 donors presented uneventful postoperative courses, 8 experienced minor complications and 2, major complications: biliary stricture and portal vein thrombosis. Recipients complications included biliary complications (35%), acute rejection (20%), hepatitis C reactivation (20%), hepatic vein stenosis (10%), hepatic artery stenosis (5%), and hepatocellular carcinoma recurrence (5%). CONCLUSIONS: LRLT has become a standard option in adults with end-stage liver failure in our center. PMID- 16213332 TI - Donor outcomes in right lobe adult living donor liver transplantation: single center experience in Egypt. AB - INTRODUCTION AND OBJECTIVES: Living donor liver transplantation (LDLT) is an alternative source of organs for patients with end-stage liver disease (ESLD) in absence of deceased donor LT. In LDLT the greatest concern is donor safety. Our objective was to evaluate the outcome of donors after right lobe liver donation in a single LT center in Egypt. PATIENTS AND METHODS: Fifty LDL resections were performed from 2001 to 2004. The mean donor age was 29.2 +/- 6.4 years. Residual liver volume was 41.1 +/- 4.5%. Mean operative time was 560 +/- 62.2 minutes; mean ICU stay, less than 24 hours; mean hospital stay, 15.4 +/- 7.7 days; and mean follow-up period, 6 months. RESULTS: There was no mortality. The overall complication rate was 68% (34 donors). Major complications included intraoperative bleeding in one, biliary leak in two, and pneumonia in three donors. Minor complications included mild pleural effusion in 13 donors, transient ascites in 10, mild depression in 7, intra-abdominal collections in 3, and wound infections in 1 donor. Residual liver volume did not affect the complication rate. None required reoperation. Return to predonation activity occurred within 6 to 8 weeks. No liver impairment occurred during follow-up. CONCLUSION: Right lobe adult LDLT is a safe procedure with regard to donor outcome. Major complications occurred in only 10% of our series. PMID- 16213333 TI - Influence of HLA compatibility on success with living-related pediatric liver transplantation. AB - It is not clear how HLA compatibility influences acute rejection and postoperative complications in cadaveric liver transplantation. Even less is known about this factor in pediatric living-related liver transplantation (LRLT). This research assessed HLA compatibility relative to rejection rates and complications in pediatric LRLT. The study retrospectively investigated data from 14 pediatric LRLTs in which the donor and recipient HLA genotypes were determined preoperatively. Three recipients (21.4%) developed biliary complications (two biliary leakage, one bile duct stenosis). Three others (21.4%) developed vascular complications (two hepatic artery thrombosis, one hepatic artery stenosis). Eight recipients (57.1%) were diagnosed with acute rejection. The incidence of acute rejection was not correlated with the number of HLA mismatches (P > .05), or with the number of HLA class I mismatches (P > .05); however, it was negatively correlated with number of HLA class II mismatches (P = .02). Arterial and biliary complications were not correlated with any of these categories of HLA compatibility. In conclusion, the data from this small group of patients provided no evidence that closeness of donor-recipient HLA matching influences outcome in pediatric LRLT. PMID- 16213334 TI - Liver transplantation and tacrolimus monotherapy for hepatocellular carcinoma with expanded criteria. AB - Liver transplantation is the only curative treatment option for patients with cirrhosis and unresectable hepatocellular carcinoma (HCC) without extrahepatic dissemination. Criteria for transplantation in HCC are controversial. In this study, we evaluate the early results of liver transplantation for unresectable HCC. Between 2003 and 2004, 10 patients (three woman, seven men; aged 1.1 to 64 years) with occult or incidental HCC underwent liver transplantation. The inclusion criteria (independent of tumor size and number of tumor nodules) were: no invasion of major vascular structures and no evidence of extrahepatic disease, including that based on hilar lymph node biopsy and cytopathological examination of intraperitoneal fluid. Eight patients (80%) received tacrolimus and two patients (20%) received rapamycin monotherapy with early withdrawal of the corticosteroid. Four patients had neoadjuvant chemoembolization before transplantation. None of the patients received adjuvant chemotherapy. Two patients with hepatitis B virus cirrhosis underwent antiviral prophylaxis with anti-HBs antibody and lamivudine. During follow-up (range, 8 to 19 months), all patients did well with excellent graft function. There was no evidence of tumor recurrence on imaging studies, and there were no elevations in alpha fetoprotein or carcinoembryonic antigen levels. Low-dose immunosuppression and expanded criteria for liver transplantation for HCC appear to have beneficial effects on disease recurrence and patient outcomes, especially in regard to living donation. PMID- 16213336 TI - Survival outcome after hepatic retransplantation for hepatitis C virus-positive and -negative recipients. AB - INTRODUCTION: Hepatitis C virus (HCV)-related liver disease is the most common indication for liver transplantation in the United States. Recurrence of HCV infection in these recipients is almost uniform. The currently available antiviral treatment is known to cause significant side effects, and the rate of sustained viral response is low. There is still controversy about whether such patients should undergo subsequent transplantations for HCV disease. This study compared outcomes for hepatic retransplantation performed in HCV(+) and HCV(-) recipients at a single center. PATIENTS AND METHODS: From December 1994 through November 2003, 68 patients at our institution received a second liver allograft. Nineteen of the recipients were HCV(+) (group A) and 49 were HCV(-) (group B). All patients were followed until January 2004. The mean follow-up time after initial retransplantation was 37 +/- 29 months. Patient and graft survival for the two groups were compared. RESULTS: Seven recipients in group A (36.8%) and 22 recipients in group B (44.9%) died during follow-up. The actuarial 3-year patient survival after initial retransplantation for groups A and B were 61.7% and 51.6%, respectively. Nine patients required a second retransplantation, 3 (15.8%) in group A and 6 (12.2%) in group B. The actuarial 3-year graft survival from initial retransplantation for groups A and B were 56.3% and 45.7%, respectively. CONCLUSION: We observed slightly better patient and graft survivals at 3 years from initial retransplantation in HCV(+) recipients compared to HCV(-) recipients. This may be due to younger donor age and better selection of HCV(+) recipients in this series. PMID- 16213335 TI - Results of liver transplantation: analysis of 140 cases at a single center. AB - The Shiraz Organ Transplant Center in southern Iran has been performing all liver transplantations in Iran and certain neighboring countries for 12 years. This study evaluated the 140 operations performed from April 1993 through November 2004. Sixty-one percent of the recipients were men and 39% were women. The average recipient age was 29.9 +/- 14.0 years. One hundred twenty-eight patients has a full-size cadaveric transplant. Most frequent causes of cirrhosis were cryptogenic and viral. An acute rejection episode occurred in 47.5% of cases, and two episodes in 8%. Most frequent short-term complications included respiratory, neurologic, and biliary problems. The 1-, 2-, and 3-year patient survival rates were 92%, 89%, and 85%, respectively. The experience that the Shiraz Organ Transplant Center has had with liver transplantation indicated success comparable to that noted in other reports. The calculated trend suggests that a goal of 100 transplantations for 2005 is within reach. PMID- 16213337 TI - Transplantation of liver grafts from older donors: impact on recipients with hepatitis C virus infection. AB - INTRODUCTION: Older donor allografts are being accepted for liver transplantation (LTx) due to shortage of organs. Hepatitis C virus (HCV) infection-related disease is presently the most common indication of LT in the United States. We studied the impact of donor age on patient and graft survivals in patients with HCV infection. PATIENTS AND METHODS: One hundred fifty four consecutive HCV(+) LTx recipients (117 men, 37 women) were studied. The mean follow-up period was 41.0 +/- 30.2 months. The population was divided into four groups according to donor age: group I (< or =20 years); group II (21 to 40 years); group III (41 to 60 years); group IV (>60 years). RESULTS: Thirty-two (20.8%) patients died during follow-up and 16 patients (10.4%) required retransplantation. The actuarial 7 year patient survivals for groups I, II, III, and IV were 87.1%, 73.7%, 69.3%, and 68.5%, respectively (P = .4). Patient survivals for donor age groups III + IV (n = 95) and groups I + II (n = 59) were 68.9% and 77.2%, respectively (P = .19). The 7-year graft survivals for groups I, II, III, and IV were 82.7%, 71.8%, 65.8%, and 62.5%, respectively (P = .17). Graft survivals for groups III + IV and groups I + II were 58.4% and 76.2%, respectively (P = .03). CONCLUSION: Patient and graft survivals for HCV-positive liver transplant recipients in this study decreased progressively as the donor age increased. Patient and graft survivals were best for group I recipients. There were significant differences in graft survivals when recipients were grouped with a cutoff donor age of 40 years. PMID- 16213339 TI - Pulse contour cardiac output system use in pediatric orthotopic liver transplantation: preliminary report of nine patients. AB - Anesthetic management of orthotopic liver transplantation (OLT) in pediatric patients is challenging in terms of intraoperative bleeding, fluid management, and hemodynamic monitoring. The pulse contour cardiac output (PiCCO) system, a relatively new device based on the single-indicator transaortic thermodilution technique, may be useful for intraoperative hemodynamic monitoring in pediatric patients. This is a preliminary report of PiCCO use in nine children (aged 9.8 +/ 4.7 years) undergoing OLT. Hemodynamic volumetric parameters monitored by the PiCCO system were mean arterial pressure (MAP), cardiac index (CI), intrathoracic blood volume index (ITBVI), extravascular lung water index (EVLWI), systemic vascular resistance index (SVRI), and stroke volume variability (SVV). All parameters were recorded at anesthesia induction (T0), at the end of the anhepatic phase (Tanhepatic), and at the end of operation (Tend). The PiCCO system revealed similar MAP, CI, EVLWI, SVV, and SVRI values at all measurement intervals. Despite similar central venous pressure measurements, ITBVI values indicated significantly lower values at Tanhepatic than at T0 (627 +/- 160 mL/m2 and 751 +/- 151 mL/m2, respectively, P = .013). There were no PiCCO catheter related complications in any patient. These findings demonstrate that the PiCCO system is a safe, continuous, multiparameter invasive monitoring device for use in pediatric patients undergoing OLT. This system may provide valuable data during pediatric OLT and appears to be a promising monitoring tool in these patients. PMID- 16213338 TI - Change in oral absorption of tacrolimus in a liver transplant recipient after reversal of jejunoileal bypass: case report. AB - INTRODUCTION: Jejunoileal bypass (JIB) was, at one time, a popular surgical technique for the treatment of morbid obesity. However, this operation was also associated with major complications. Consequently, many such procedures were eventually reversed. One of the most serious of these complications was liver failure. For those patients who developed cirrhosis, liver transplantation was one therapeutic alternative. Tacrolimus is one of the primary immunosuppressive agents used in liver transplantation. It is effective to prevent acute rejection episodes, but shows a narrow therapeutic index and can cause nephrotoxicity and neurotoxicity. This report describes the change in tacrolimus absorption that was observed after JIB reversal in a 57-year-old female liver transplant recipient. RESULTS: Prior to JIB reversal, the mean tacrolimus dose was 7 mg twice daily with a whole-blood tacrolimus concentration ranging from 5.2 to 6.4 ng/mL. There was no appreciable peak in tacrolimus concentration, and the area under the concentration-time curve (AUC) was 10.9 ng/mL/h. After reversal, the daily tacrolimus dose was decreased to 5 mg twice daily, with a now-discernable peak concentration at 3 hours postdose. Furthermore, the AUC increased 90% to 20.7 ng/mL/h. CONCLUSION: After JIB reversal, the patient showed higher systemic levels of tacrolimus and required lower steady-state doses. It is therefore imperative that such patients be monitored closely to avoid tacrolimus-related toxicity. PMID- 16213340 TI - Internal jugular versus subclavian vein catheterization for central venous catheterization in orthotopic liver transplantation. AB - The aim of this study was to compare incidence rates of mechanical and infectious complications associated with central venous catheterization via the internal jugular vein (IJV) versus the subclavian vein (SV) among 45 consecutive patients undergoing orthotopic liver transplantation (OLT) between January 2000 and June 2004. The subjects were divided into two groups according to the site of central venous catheterization (IJV or SV). We recorded each patient's physical characteristics, international normalized ratio (INR), partial thromboplastin time, platelet levels, number of puncture attempts, success/failure of central venous catheterization, duration of catheter placement, occurrence of catheter tip misplacement, arterial puncture, incidence of hematoma or pneumothorax, catheter-related infection, or bacterial colonization of the catheter. Senior staff anesthesiologists performed 22 SV and 23 IJV catheterizations for the 45 OLT procedures. The SV and IVJ groups both had minor coagulation abnormalities with slightly increased INR values at the time of catheterization. There were no significant differences between the groups with respect to success of central venous catheterization (100% for both), numbers of attempted punctures, duration of catheter placement, and incidence rates of mechanical and infectious complications. Both groups showed high frequencies of catheter tip misplacement, with right atrium as the site of misplacement in all cases. Two patients in the IJV group (8.7%) developed hematomas after accidental carotid artery puncture. The results suggest that, when performed by experienced anesthesiologists, central venous catheterization via the SV is an acceptable alternative to IJV catheterization for patients undergoing OLT. PMID- 16213341 TI - Biliary complications and their management in pediatric liver transplantations: one center's experience. AB - Following living-donor liver transplantation, biliary complications are more prevalent among pediatric patients (<18 years old), with reported rates varying between 15% and 30%. In this study, we retrospectively analyzed biliary complications observed in 21 pediatric liver transplant patients (16 boys [76.2%], 5 girls [23.8%] of ages 1 to 18 years (mean, 8.3 +/- 5.05 years) between September 2001 and June 2004. Biliary reconstruction was accomplished via a duct to-duct anastomosis in 12 (57.1%) and a Roux-en-Y hepaticojejunostomy in 9 (42.9%) patients. Postoperative biliary complications were encountered in six (28.5%) patients. Four of the biliary leaks were from a duct-to-duct anastomosis and two from an hepaticojejunostomy. One (4.7%) patient who experienced biliary leakage after a duct-to-duct anastomosis developed stenosis after the leak healed; five (23.8%) had the leakage treated successfully. One patient had biliary leakage from the duct-to-duct anastomosis subsequent to a hepatic artery thrombosis. All patients with biliary leakage were treated without surgery. Mean follow-up time was 10.2 +/- 9.6 months (range, 1 to 26 months). Three patients died during follow-up; however, these deaths were not related to the biliary complications. Interventional radiologic approaches are effective to biliary complications, even when the anastomoses are heavily disrupted. In cases of biliary complication, percutaneous combined with internal drainage may prevent biliary sepsis and provide long-term patency. PMID- 16213342 TI - Biliary tract complications after liver transplantation in a single center. AB - Biliary tract complications, which occur in 5.8% to 24.5% of adult liver transplant recipients, remain one of the most common problems following transplantation. The aim of this study was to determine the incidence of biliary complications and analyze methods of treatment. From 1993 to 2004, 14 cases (10%) among 140 patients who had undergone liver transplantation developed biliary complications, third to respiratory and neurologic complications. In addition to biliary leakage in six cases, obstruction/stenosis occurred in three cases. One case of biliary fistula and one vanishing bile duct syndrome were observed. There was no death or need for retransplantation; all cases were treated surgically without recurrence. Biliary complications remain an important problem in liver transplantation. Endoscopic and radiologic management are effective in the majority of cases. Surgical intervention is obligatory and safe in selected cases. PMID- 16213343 TI - Incidence and management of biliary complications after orthotopic liver transplantation: ten years' experience at King Fahad National Guard Hospital. AB - BACKGROUND: Despite technical modifications and application of various surgical techniques, biliary tract complications remain a major source of morbidity after orthotopic liver transplantation. We sought to assess the incidence and management of biliary complications at a single liver transplant unit. METHODS: Among 184 consecutive deceased donor liver transplants performed between February 1994 and July 2004, 66 were female patients and 118 male patients of age range 2(1/2) to 69 years. We retrospectively reviewed the data regarding biliary complications in liver transplant recipients, after 115 duct-to-duct anastomoses and 65 hepaticojejunostomy. We analyzed the incidence and type of biliary complications, management sequence, and success rate. We analyzed the correlation between the modality of biliary reconstruction and the type/incidence of biliary complications. RESULTS: Thirty-two patients developed biliary complications, giving an overall incidence of 17.4%. There was a higher incidence of complications among patients in the hepaticojejunostomy group (21.5%) than the duct-to-duct technique (15.1%). Bile leakage occurred in 12 patients, including eight successful cases (66.6%) of endoscopic stent insertion/radiological techniques and surgery in four cases (33.3%). Among the 12 patients with initial leaks, six developed a subsequent stricture (50%). There were 26 cases of biliary stricture, including 22 (84.6%) who were initially managed using nonsurgical techniques with a success rate of 59%. CONCLUSION: Biliary complications remain an important cause of morbidity after orthotopic liver transplantation. They can usually be managed percutaneously or endoscopically; however, tight strictures and major leaks frequently required surgical intervention. PMID- 16213344 TI - Does valganciclovir hydrochloride (valcyte) provide effective prophylaxis against cytomegalovirus infection in liver transplant recipients? AB - INTRODUCTION: Cytomegalovirus (CMV) infection after solid organ transplantation is one of the most common viral infections, causing significant morbidity and mortality if not treated promptly. Ganciclovir has proven to be effective for the prophylaxis and treatment of CMV. However, oral absorption of ganciclovir is poor. Recently, oral administration of valganciclovir hydrochloride (Valcyte) has been observed to display 10-fold better absorption than oral ganciclovir. Valganciclovir has increasingly been used as prophylaxis against CMV after solid organ transplantation. The purpose of this study was to examine the efficacy of valganciclovir prophylaxis therapy after primary liver transplantation. PATIENTS AND METHODS: Between July 2001 and May 2003, 203 consecutive liver transplant recipients, including 129 men and 74 women of overall mean age 53 +/- 11 years, received valganciclovir (900 mg/d or 450 mg every other day depending on renal function) for 3 to 6 months after primary liver transplantation. All patients were followed up for a minimum of 6 months. Mean follow-up was 19 +/- 5.8 months. CMV DNA in peripheral blood was tested using polymerase chain reaction (PCR) amplification. Symptomatic CMV was stratified according to the CMV immunoglobulin (Ig)G status of the donor and recipient at the time of liver transplantation. Donors and recipients were classified preoperatively into groups according to the presence or absence of CMV as follows: group 1 (n = 73; donor CMV+, recipient CMV+); group 2 (n = 41; donor CMV-, recipient CMV+); group 3 (n = 54; donor CMV+, recipient CMV-; high-risk group); and group 4 (n = 35; donor CMV-, recipient CMV ). RESULTS: Twenty-nine patients (14.3%) developed symptomatic CMV disease at 169 +/- 117 days after liver transplantation: group 1, 16.4% versus group 2, 7.3% versus group 3, 25.9% versus group 4, 0%. Of these patients, 5 also had invasive CMV on liver biopsy, which was performed owing to abnormal liver functions. All 29 patients were treated with intravenous ganciclovir. One patient died owing to disseminated CMV, whereas the remaining 28 patients responded to treatment. Interestingly, 8 patients, including 1 who had invasive CMV hepatitis, developed symptomatic CMV within 90 days of liver transplantation even while on prophylactic valganciclovir. CONCLUSION: Valganciclovir failed to provide adequate prophylaxis following liver transplantation in our patients. The overall rate of CMV in seropositive donors and/or recipients was 17%, and in the high risk group was 26%. Further prospective studies with measurement of ganciclovir concentrations are needed to elucidate the reasons for this unexpected failure. PMID- 16213345 TI - Use of hepatitis B core antibody-positive liver allograft in hepatitis C virus positive and -negative recipients with use of short course of hepatitis B immunoglobulin and Lamivudine. AB - INTRODUCTION: With the shortage of donor organs, increasing number of hepatitis B core antibody (HBcAb)-positive [HBcAb(+)] liver allografts are being used for liver transplantation (LTx) in patients who are HBcab-negative [HBsAb(-)]. This study was aimed at assessing outcomes for hepatitis C virus (HCV)-positive [HCV(+)] and HCV-negative [HCV(-)] patients who received HBcAb(+) liver grafts from deceased donors and also received a short course of hepatitis B immunoglobulin (HBIg) with long-term lamivudine therapy after LTx. MATERIALS AND METHODS: From February 1995 through February 2003, 28 patients (mean age 53.8 +/- 10.2 years, 19 men and nine women, 16 HCV[-]; 12 HCV[+]) received HbcAb(+) liver allografts. All recipients received a short course of HBIg prophylaxis (10,000 units/day for 4 days) and long-term lamivudine 100 mg/d after LTx in addition to a tacrolimus-based immunosuppressive regimen. RESULTS: Seven (25%) of the 28 recipients died during follow-up and three recipients required retransplantation. Three recipients (10.7%) developed HBV infection during follow-up, one of whom died 36 months after LTx and the other two had YMDD mutant HBV. The overall 6 year actuarial patient survival after transplantation was 74.4% and those for HCV(-) and HCV(+) recipients were 81.3% and 66.6%, respectively (P = .46). The overall 6-year actuarial graft survival was 63.9% and those for HCV(+) and HCV(-) recipients were 68.8% and 57.1%, respectively (P = .6). CONCLUSION: We conclude that HBcAb(+) liver grafts can be used for both HCV(+) patients and HCV(-) patients who are critically ill, have early hepatocellular carcinoma, or have been exposed to HBV in the past. A short course of HBIg-lamivudine combination therapy provides effective prophylaxis against HBV infection in 89% of recipients of HBcAb(+) grafts. PMID- 16213346 TI - Ribavirin levels in post liver transplant patients treated for recurrent hepatitis C viral infection. AB - Hepatitis C virus (HCV) infection is the most common indication for liver transplantation (LTx) in the United States. Ribavirin with pegylated interferon is the only treatment option for HCV recurrence in post-LTx patients. In clinical practice, for more than 50% of patients, ribavirin dose needs to be modified. AIM: The aim of this study was to examine the role of ribavirin level and its relevance in the management of post-LTx patients in terms of renal dysfunction, efficacy, toxicity, and potential drug interactions. PATIENTS AND METHODS: Thirty four blood samples were available from 22 post-LTx patients. Ribavirin concentrations in plasma (all samples) and whole blood concentrations (16 samples) were examined. The dose of ribavirin ranged from 400 mg/d to 1000 mg/d, but concentrations were normalized to 800 mg/d. RESULTS: There was a wide variation in plasma concentration of ribavirin, ranging from 1.8 to 122.1 mg/mL. The concentrations were similar in whole blood and plasma. Dose-normalized concentration with creatinine clearance below 70 mL/min were significantly higher when compared with creatinine clearance above 70 mL/min (P = .015). Eleven patients required erythropoietin; their mean ribavirin dosage was higher but mean ribavirin concentration was lower compared to the 11 patients who did not require erythropoietin factor. There was no difference in mean ribavirin concentration in patients who cleared the virus (n = 7) compared and who did not clear the virus (n = 9). Three patients were on nucleoside reverse transcriptase inhibitors (NRTI) had significantly higher concentration (mean 87.1 microg/mL) compared to those who did not receive NRTI (mean 34.4 microg/mL, P = .00) CONCLUSION: Ribavirin concentration in plasma and whole blood were similar, with a wide variation. Patients with impaired renal function and those who were on NRTI had significantly higher concentrations of ribavirin. The ribavirin concentrations did not predict either the clearance of HCV RNA or the need for erythropoitin factor. PMID- 16213347 TI - Neurological complications of first 100 orthotopic liver transplantation patients in southern Iran. AB - Neurological complications which are responsible for significant mortality and morbidity after orthotopic liver transplantation have been reported in 8.3% to 47% of cases in various series. This prospective study was performed to investigate the prevalence and characteristics of neurological complications in the first 100 patients who have undergone orthotopic liver transplantation from 1993 to 2004. Neurological symptoms and signs as well as routine laboratory tests, including complete blood cell counts, electrolyte levels, drug levels, microbiologic and serological studies, brain computed tomography scans, magnetic resonance imaging, and electrodiagnostic studies were reviewed in all patients. Follow-up periods were from 2 months to 10 years. Nineteen patients of mean age of 34.9 years developed neurological complications after orthotopic liver transplantation. The most common neurological symptoms and signs were confusion (42.1% of cases with neurological complications), convulsions (36.8%), and hallucinations (31.6%). Hepatic encephalopathy (31.6%) and drug toxicity (26.2%) were the most common neurological syndromes. The mortality rate was significantly higher among patients with neurological complications. Compared to other centers, neurological complications were less common in our center. PMID- 16213348 TI - Vascular complications after liver transplantation in pediatric patients. AB - Vascular complications are the major cause of morbidity and mortality after liver transplantation, particularly in pediatric patients, owing to their smaller vascular diameters. Between September 2001 and June 2004, among 21 (16 boys and 5 girls) pediatric liver transplantations of mean age 8.3 +/- 5.1 years, hepatic arterial thrombosis (HAT) was diagnosed in 2 (9.5%) patients, and hepatic arterial stenosis (HAS) in 4 (19.4%). Vascular patency was evaluated with Doppler ultrasonography every 12 hours in the first postoperative week and daily in the second postoperative week. When occlusion was suspected, conventional angiography was performed. Thrombectomy was performed in one patient, and thrombectomy and reanastomosis were performed in another patient with HAT. Two patients with HAS were treated with balloon angioplasty. A third patient was treated with balloon angioplasty and endoluminal stent placement at the same time. The last patient with HAS had an intimate dissection, which occurred 24 hours after balloon angioplasty, that was treated with subsequent endoluminal stent placement. Mean follow-up for the patients with vascular complications was 9.5 +/- 5.7 months (range, 4 to 18 months). The overall mortality rate was 14.1% (3/21); however, no deaths were caused by vascular complication. Routine Doppler ultrasonographic evaluation is an effective choice for diagnosing vascular complications seen after liver transplantation. Immediate surgical intervention is required for acute vascular complications, whereas late complications may be treated with balloon angioplasty and/or endoluminal stent placement. PMID- 16213349 TI - Kidney-pancreas transplantation: single-center experience at a university hospital in Turkey. AB - INTRODUCTION: One treatment option for patients with type 1 diabetes mellitus with end-stage nephropathy is combined pancreas-kidney transplantation, which can be performed either simultaneously (SPK) or following kidney transplantation (PAK). PATIENTS AND METHODS: Between February 2003 and November 2004, 14 patients, including 10 males and 4 females of overall mean age of 31.3 +/- 6.1 years (range, 23-44 years), presented with end-stage renal disease secondary to type 1 diabetes mellitus. Five patients (35.7%) received SPK; 7 patients (50%) received PAK; and 2 patients (14.3%) received simultaneous pancreas and living related kidney (SPLK) transplantations. RESULTS: Two among 14 pancreas grafts were lost in the early postoperative period secondary to venous thrombosis despite anticoagulation including 1 with poor portal drainage. Insulin therapy was reinitiated in 1 patient after a second rejection episode in the seventh postoperative month. By the ninth median follow-up month (range, 1-21 months), all kidney grafts were functioning. CONCLUSION: Our single-center short-term experience with 14 consecutive kidney-pancreas transplantations suggests that while the pancreas transplant is effective and safe to reestablish normoglycemia, this transplant creates additional surgical and immunosuppressive stresses on the patient. PMID- 16213350 TI - Risk of viral transmission via bone marrow progenitor cells versus umbilical cord blood hematopoietic stem cells in bone marrow transplantation. AB - Hematopoietic stem cell transplantation (HSCT) is the treatment of choice for children and certain adults with malignant and nonmalignant hematologic disease. Since viral infections are the major problem, this study examined those that might potentially be transmitted to HSCT recipients via bone marrow (BM) versus umbilical cord blood (UCB). BM progenitor cells, peripheral blood leukocytes, and plasma samples were collected from 30 allogenic BM donors. Umbilical cord blood hematopoietic stem cells and plasma samples were also collected from 34 UCB donors. Viral DNA extracted and purified from collected specimens was processed using nested polymerase chain reactions (PCR) to detect human parvovirus B19 (HPV B19), human herpesvirus-6 (HHV-6), varicella-zoster virus (VZV), human cytomegalovirus (HCMV), and Epstein-Barr virus (EBV). The prevalences of HCMV DNA in collected BM progenitor cells versus UCB hematopoietic stem cells were 73% versus 23%, respectively. Conversely, HHV-6 DNA was not detected in any collected specimen by simple PCR. Distribution of the other investigated virus DNAs except EBV DNA was similar in specimens collected from both groups. EBV DNA was not determined in UCB hematopoietic stem cells. The results indicate that the risk of viral transmission to BM transplant recipients via UCB hematopoietic stem cells is less than that with BM progenitor cells. PMID- 16213351 TI - Antineutrophil cytoplasmic antibody-associated glomerulonephritis in chronic graft-versus-host disease after allogenic hematopoietic stem cell transplantation. AB - Graft-versus-host disease (GVHD) is one of the most frequent complications that occur after hematopoietic stem cell transplantation (HSCT). Recently, renal involvement, including membranous nephropathy, focal segmental glomerulosclerosis, and minimal change disease, has been described as a manifestation of chronic GVHD. This case report describes a patient who developed antineutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis after HSCT. Following preparation with chemotherapy, a 29-year-old man with chronic myeloid leukemia underwent allogenic peripheral blood stem cell (PBSC) transplantation, after which first acute and then chronic GVHD developed. Treatment with prednisone resulted in improvement in the patient's GVHD. After the termination of steroid therapy and about 10 months after PBSC transplantation, nephritic syndrome appeared and the patient's serum creatinine value increased to 1.7 mg/dL. Laboratory evaluation revealed perinuclear antineutrophilic cytoplasmic antibody (p-ANCA) in the serum. Histological examination of renal biopsy tissue showed focal segmental proliferative glomerulonephritis with glomerulosclerosis in 20% of available glomeruli, large cellular crescents in 6% of glomeruli, and no staining of immunoglobulins or complement along the capillary walls. Electron microscopy revealed no immune deposits. After treatment with prednisone 60 mg/d, diltiazem 120 mg/d, and enalapril 10 mg/d, the proteinuria gradually decreased, and p-ANCA was undetectable. These findings suggest that in this patient the ANCA-associated glomerulonephritis was associated with renal involvement that occurred during the course of chronic GVHD. PMID- 16213352 TI - Low-dose histidine-tryptophan-ketoglutarate solution for myocardial protection. AB - The effect of histidine-tryptophan-ketoglutarate (HTK) solution for myocardial protection has been shown in experimental and clinical studies using long ischemic times and high dosages. In our study we compared myocardial protection in isolated coronary bypass with a short period of ischemia using low dosage HTK and cold crystalloid cardioplegia. Each group contained 21 coronary artery disease patients. Cardioplegic solutions were administered antegrade in 10 to 15 mL/kg in one shot. This dosage of HTK was lower than that mentioned in the literature. We measured malondialdehyde, lactate, creatine kinase, creatine kinase-MB, and troponin-I levels. Aortic clamping time in the HTK group 33.9 +/- 8.2 minutes, versus 36.2 +/- 11.3 minutes in the crystalloid cardioplegia group (P > .05). Levels of creatine kinase and malondialdehyde were lower in HTK group at 24 hours and 2 minutes, respectively. Lactate levels were lower in the crystalloid cardioplegia group at 2 minutes in the coronary sinus serum sample, but there were no statistically differences among ischemic serum markers in both groups. Only intervals between aortic clamping and cardiac arrest were statistically meaningful (HTK 63.3 +/- 14.7 seconds versus crystalloid cardioplegia 53.6 +/- 15.6 seconds, P = .044). Our study shows that use of low dose HTK for short clamping time operations is as successful for myocardial protection as crystalloid cardioplegia. Longer times for fibrillation can be explained with the low levels of potassium in HTK solution, but this length did not cause a biochemical or clinical difference. PMID- 16213353 TI - Cobalt-protoporphyrin induced heme oxygenase overexpression and its impact on liver regeneration. AB - Cobalt-protoporphyrin (CoPP)-dependent induction of heme oxygenase (HO)-1 has been shown to protect from ischemia-reperfusion injury, which remains a major source of graft loss after liver transplantation. The impact of HO-1 on liver regeneration, especially in reduced-size grafts, has not yet been evaluated. Using an experimental model, we investigated HO-1 induction by CoPP treatment on postoperative recovery of ischemically injured livers following partial (70%) hepatectomy. Wistar rats underwent partial hepatectomy under temporary inflow occlusion (30 minutes). One group of animals received CoPP (5 mg/kg body weight i.p.) 24 hours prior to surgery to induce high levels of HO-1 at the time of surgery, and the second group served as nontreated controls. At postoperative days 1, 4, 7, and 10, animals were exsanguinated, and blood and liver samples were stored for enzymatic (serum AST and ALT levels) and histologic (mitotic index) analyses (n = 5 each day). Additionally, postoperative body weight and weight of the remnant liver were measured. Although serum AST and ALT levels as well as remnant liver weight were comparable between both groups, CoPP-treated animals recovered from surgery more quickly as indicated by postoperative body weight. Moreover, the number of mitotic cells was significantly increased in this group at day 1 (33 +/- 5 versus 20 +/- 5 per 2000 hepatocytes) as compared with nontreated animals. Liver regeneration of ischemically injured livers following partial hepatectomy was improved by HO-1 overexpression following preoperative CoPP administration. Thus, it is conceivable that prevention of ischemia reperfusion injury by HO-1 overexpression also might be beneficial for reduced size liver grafts without affecting their proliferative capacity. PMID- 16213355 TI - The human Penumbra gene is mapped to a region on chromosome 7 frequently deleted in myeloid malignancies. AB - We previously cloned the murine Penumbra gene based on its differential expression in proerythroblasts/erythroblasts. Subsequently, we identified human Penumbra cDNA from a human bone marrow cDNA library and the human Penumbra gene from a BAC library. Penumbra is a new member of the tetraspanin protein family and exhibits growth-suppressive activity in vitro. In this study, we designed a human Penumbra probe contig and used fluorescent in situ hybridization (FISH) to analyze seven cases of myeloid malignancies with 7q deletions. Five patients with cytogenetic deletions involving 7q31.2 approximately q32 also showed deletions of Penumbra by FISH; these were not present in two patients with cytogenetic deletions not involving 7q31.2 approximately q32. Our findings provide the first FISH evidence supporting the mapping of human Penumbra to 7q31.2 approximately q32 and demonstrate the potential of the Penumbra probe in the detection of 7q31 approximately q32-related deletions in myeloid malignancies. PMID- 16213354 TI - Effect of cyclosporine, mycophenolate mofetil, and their combination with steroids on apoptosis in a human cultured monocytic U937 cell line. AB - Transplant patient plasma produces an increased rate of mononuclear cell apoptosis despite a normal serum creatinine value. Immunosuppressive medications may be one factor that causes an altered apoptotic pattern. We evaluated the in vitro effects of various doses of cyclosporine, mycophenolate mofetil, and steroids on apoptosis of a cultured human monocytic U937 cell line, using estimates by fluorescence microscopy and annexin V assays. Increasing cyclosporine concentrations (100 to 800 ng/mL) progressively increased apoptosis rates (16% to 32%). The combination of steroid (0.01 microg/mL) and cyclosporine increased the apoptosis rate to 45%. Mycophenolate mofetil alone (0.3 microg/mL) led to an apoptosis rate of 34%. Therapeutic levels of mycophenolate mofetil from 3 to 7 microg/mL led to apoptosis rates from 56% to 67%. The combination of cyclosporine, steroid, and mycophenolate mofetil increased the rate of apoptosis to 95%. Immunosuppressive therapy may contribute to the high rate of apoptosis observed among mononuclear cells of transplanted patients. This effect may alter patient susceptibility to infections and contribute to a unique mechanism of immunosuppression. PMID- 16213356 TI - Characterization of genomic instability in ulcerative colitis neoplasia leads to discovery of putative tumor suppressor regions. AB - Ulcerative colitis (UC) is an inflammatory disease of the colon that is associated with increased risk of colorectal cancer associated with genomic instability. We have previously demonstrated that genomic instability is present in UC patients with colonic neoplasia, and hypothesized that the chromosomal alterations may be taking place in regions that are susceptible to mutation or that provide a growth advantage to a cell undergoing neoplastic transformation. In this study, we used two polymerase chain reaction (PCR)-based DNA fingerprinting techniques (arbitrarily primed PCR and inter-simple-sequence repeat PCR) to study the process of genomic instability. The two techniques of DNA fingerprinting cross-validate the instability observed in these studies. We analyzed the molecular basis of 10 commonly altered DNA bands obtained from DNA fingerprints of biopsies from various histologic grades of UC patients with dysplasia or cancer (UC Progressors). We determined that the band changes in the fingerprint truly represent changes in DNA sequence, and that the fingerprinting provides highly reproducible results. Furthermore, our investigation revealed that 40% of alterations involve repetitive sequences. Two frequently deleted sequences in 6q27 and 2q14 were studied further because they were frequently abnormal in the dysplastic and nondysplastic tissue of UC Progressors. The losses from 6q27 and 2q14 were confirmed by loss of heterozygosity and real-time PCR analysis. Both of these regions in chromosomes 6 and 2 are surrounded by highly repetitive and mobile LINE-1 elements, possibly making the region susceptible to mutational change. These regions were affected (lost) in UC Progressors but not in UC patients who were neoplasia free. Loss of heterozygosity at 6q27 has been described in ovarian and other cancers, while the 2q14 region has been implicated in prostate and sporadic colon cancers. Both regions are likely to contain tumor suppressor genes. In conclusion, the genomic instability in UC Progressors can occur in regions that are susceptible to change and are locations of putative tumor-suppressor genes. PMID- 16213357 TI - Chromosomal aberrations in follicular non-Hodgkin lymphomas of Japanese patients, detected with comparative genomic hybridization and polymerase chain reaction analysis. AB - The t(14;18)(q32.3;q21), hallmark of follicular lymphoma (FL), is less frequently observed in Asian patients than in Westerners. Little is known about additional chromosomal aberrations in Asian FL patients. We applied comparative genomic hybridization (CGH) to screen for genomic imbalances in 32 biopsy samples from 23 Japanese patients with nodal FL. The t(14;18) was assessed with polymerase chain reaction (PCR) using primers for the major (MBR), 3'MBR, minor (mcr), 5'mcr, and intermediate breakpoint cluster regions (icr). In 17 of 23 patients, CGH analyses detected genomic imbalances. Gains frequently affected chromosomes 18p (6 of 23), X (5 of 23), 5 (4 of 23), 12 (4 of 23), 2 (3 of 23), and 16 (3 of 23). The only recurrent loss affected chromosome 6q (2 of 23). A t(14;18) was detected in 13 of 23 patients (56.5%). Breakpoints were located in the MBR (10 patients), in the 3'MBR, in the mcr, and in the icr (1 patient each). The frequency of aberrations detected by CGH as well as relapse-free survival were not distinctly different between patients with and without a t(14;18). In summary, no significant difference in the overall frequency of aberrations compared with Westerners was found. Despite the extended primer set used, the frequency of PCR-detected t(14;18) remained low. Additional studies are required to assess the reasons for geographic variation in FL. PMID- 16213358 TI - Alterations of 9p in squamous cell carcinoma and adenocarcinoma of the lung: association with smoking, TP53, and survival. AB - Tobacco smoke is well recognized as the major etiological contributor to lung cancer, yet the relationship between tobacco smoke exposure and a specific pattern of molecular abnormalities at somatic loci is less well characterized. We analyzed 100 primary tumors from patients undergoing surgical resection of squamous cell carcinoma and adenocarcinoma of the lung for loss of heterozygosity (LOH) and homozygous deletions at two microsatellite markers in a recombinogenic region of 9p13. We describe the relationship of alterations at these markers with tumor characteristics (both clinical and molecular), patient demographics, survival, and measures of tobacco-smoke exposure. Homozygous deletions in this region occurred in 25% (21/85) and LOH in 33% (28/85) of informative tumors examined. These alterations occurred more often in tumors with intense TP53 protein staining by immunohistochemistry, suggesting that inactivation of the TP53 pathway may contribute to these LOH events. Duration of smoking was greatest in patients with the homozygous deletion, intermediate in patients with LOH, and shortest in patients whose tumor did not demonstrate loss in these markers. Unexpectedly, LOH at 9p13 was a significant predictor of improved survival in patients, while the homozygous deletion was associated with the poorest patient survival. Together, these results suggest that TP53 alteration and long-term tobacco smoke exposure may contribute to genetic alterations at 9p13, and that the mechanism and biologic consequences of allele loss reflect individual biologic differences that determine the extent of loss (LOH or homozygous deletion), such that those patients with the deletion of this region face a more aggressive and deadly disease. PMID- 16213359 TI - 3'CBFbeta deletion associated with inv(16) in acute myeloid leukemia. AB - Recent reports have shown that concomitant submicroscopic deletions can occur in association with chromosomal translocations/inversions in several leukemia subtypes. Detectable by fluorescence in situ hybridization (FISH), these losses of sequence include deletion of the 5' region of the ABL gene and the 3' region of BCR in chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL), as well as the 5' region of ETO in acute myeloid leukemia (AML) French-American British type M2 associated with t(8;21), 3'MLL in AML and ALL, and 3' core binding factor beta (CBFbeta) in AML associated with inv(16). While it has been widely reported that submicroscopic deletions of the derivative 9 in CML have an adverse prognostic impact, the clinical significance, if any, of deletions associated with t(8;21), inv(16)/t(16;16), or MLL rearrangement is yet to be determined. We analyzed a series of 39 patients diagnosed with AML who had cytogenetically detectable inv(16)/t(16;16) by using a FISH probe for the CBFbeta region to determine the incidence of the 3'CBFbeta deletion. Deletions were detected in three patients (8%), all associated with inv(16), bringing the number of cases reported so far to seven. The prognostic significance of this finding remains unclear. PMID- 16213361 TI - Inactivation patterns of NF2 and DAL-1/4.1B (EPB41L3) in sporadic meningioma. AB - The molecular basis of tumorigenesis and tumor progression in meningiomas is not fully understood. The neurofibromatosis 2 (NF2) locus is inactivated in 50-60% of sporadic meningiomas, but the genetic basis of sporadic meningiomas not inactivated at the NF2 locus remains unclear. Specifically, there is conflicting data regarding the role of the tumor suppressor gene DAL-1/4.1B. Using microsatellite markers, we studied 63 sporadic meningiomas to determine loss of heterozygosity (LOH) at the NF2 and DAL-1/4.1B loci. Array comparative genomic hybridization analysis of 52 of these tumors was performed to determine copy number changes on chromosomes 18 and 22. Forty-one of 62 informative tumors showed LOH at the NF2 locus (66%) while only 12 of 62 informative tumors (19%) showed LOH of DAL-1/4.1B. Eleven of 12 (92%) tumors with DAL-1/4.1B LOH also had NF2 LOH. Monosomy or large deletions of chromosomes 18 and 22 were the main mechanism for LOH in these tumors. These studies implicate the DAL-1/4.1B locus in sporadic meningiomas less commonly than reported previously, and suggest that it is a progression rather than an initiation locus. Furthermore, we found the majority of meningiomas developed monosomy rather than isodisomy at the NF2 and DAL-1/4.1B loci as the mechanism for LOH. PMID- 16213360 TI - Mutations of the FLT3 gene in adult acute myeloid leukemia: determination of incidence and identification of a novel mutation in a Thai population. AB - FLT3 mutations have been reported to be the most frequent mutation in acute myeloid leukemia (AML). No data currently exist regarding FLT3 mutations in Southeast Asian patients. In this study, the incidence and type of FLT3 mutation in a large series of Thai AML patients were determined. FLT3 internal tandem duplication (ITD) mutations were observed in 24.6%, FLT3 tyrosine kinase domain mutations in 3.1%, and dual mutations in 2.7% of 256 newly diagnosed Thai AML patients. ITD mutations were mostly restricted to the juxtamembrane domain, and the in-frame ITD length varied from 21 to 201 base pairs. Six types of point mutations were identified, including Asp835Tyr, Asp835His, Asp835Glu, Asp835Ala, Ile836, and a novel mutation, Asp835Del/Ile836Val, which resulted in the loss of aspartic acid and substitution of isoleucine by valine. A rare leukemia karyotype, trisomy 11, was found to coexist with this novel FLT3 mutation, whereas the majority of patients with FLT3 mutations had a normal karyotype. Overall, FLT3 mutation was associated with a significantly higher white blood cell count and older age than the wild-type FLT3. In conclusion, the incidence of FLT3 mutation in Thailand is as high as that of western countries. The clinical significance of the novel mutation requires further studies in a larger population. PMID- 16213362 TI - Detection of ETV6 and RUNX1 gene rearrangements using fluorescence in situ hybridization in Mexican patients with acute lymphoblastic leukemia: experience at a single institution. AB - The t(12;21) produces the gene fusion ETV6/RUNX1 and is a frequent rearrangement in childhood ALL, associated with a good prognosis. In Mexico its prevalence has not been reported. This study evaluated a group of consecutive Mexican children with newly diagnosed ALL, to detect the fusion using fluorescence in situ hybridization (FISH). Seventy-one bone marrow samples were analyzed with FISH, using ETV6/RUNX1 DNA probes. Abnormalities of ETV6, RUNX1, or both were found in 31 of the 71 (44%) patients. Six showed ETV6/RUNX1 fusion and 17, with extra RUNX1 copies, presented an additional chromosome 21 or dup(21)(q22). Five patients had structural changes in ETV6, and three patients showed extra copies of ETV6 and RUNX1 from polysomy of chromosomes 12 and 21. Our results revealed a fusion in 8.5% of the 71 cases analyzed. This frequency is lower than that observed in other populations (9.5-32%). The structural rearrangements resulting in RUNX1 extra copies were found in 9.8% of patients, which is close to the range reported (1.5-9.7%) by other authors. Due to the prevalence of RUNX1 overrepresentation in our population and its unknown prognostic significance, further studies should be conducted in consecutive children with ALL, to correlate this abnormality with the patients' follow-up. PMID- 16213363 TI - ABL1 amplification in T-cell acute lymphoblastic leukemia. AB - ABL1 amplification, due to a cryptic episomal translocation NUP214/ABL1, is a novel finding in T-cell acute lymphoblastic leukemia (ALL). Here we report on the incidence and clinical features of this genetic defect in a series of 30 consecutive adult T-cell ALL patients. Multiple copies of the ABL1 gene were detected in two patients (6.6%), one with the karyotype 46,XY,t(1;3)(p36;p21),del(6)(q23)/46,XY and the other without analyzable metaphases. Metaphase/interphase fluorescence in situ hybridization (FISH) detected multiple uncountable signals corresponding to ABL1 in mitotic cells and nuclei from both patients. In one patient, no signals corresponded with the 9p21 chromosomal region, which contains the p16INK4a gene, and in the other one signal was observed. Quantitative reverse-transcriptase polymerase chain reaction (RT PCR) demonstrated that in these patients ABL1 gene expression was 14- and 18-fold greater than in normal controls, and returned to normal levels only when complete remission was achieved. We reached the following conclusions: (1) FISH is the only technique that promptly identifies T-cell ALL patients with ABL1 amplification, (2) quick identification with FISH is fundamental in the clinic because this T-cell ALL subset is imatinib sensitive but may become resistant due to development of additional mutations, and (3) ABL1 quantitative RT-PCR may be easily applied to monitor minimal residual disease. PMID- 16213364 TI - Finer delineation and transcript map of the 7q31 locus deleted in myeloid neoplasms. AB - Acquired complete and partial deletions of chromosome 7 are associated with several malignancies. In acute myelogenous leukemia (AML) and preleukemic myelodysplasia (MDS), loss of chromosome 7 portends a poor clinical outcome. The identity of a classical leukemia suppressor gene, however, has been elusive. Previously, we defined a candidate suppressor locus of approximately 6 Mb in the 7q31 interval. Here we report an island of retention of heterozygosity within this interval in a case of monosomy 7. Allelotyping of AML cell lines revealed that ML3 and HEL cells, karyotypically diploid for chromosome 7, are hemizygous for all the 7q31 loci, implicating loss of the wild type and duplication of the remaining chromosome 7. Based on the completed genomic sequence of chromosome 7, we have generated a transcript map of the critical region of loss (between the D7S525 and D7S2502 loci). Notably, a recently characterized tumor suppressor gene, DOCK4, and an evolutionarily conserved zinc finger gene, ZNF277, localize to this interval, head to head, within <0.5 kb of each other. Thus, the reagents generated in this study will be valuable in elucidating the role of loss of 7q31 loci in the pathogenesis of AML. PMID- 16213365 TI - A der(13)t(7;13)(p13;q14) with monoallelic loss of RB1 and D13S319 in myelodysplastic syndrome. AB - Deletions or translocations of chromosome band 13q14, the locus of the retinoblastoma gene (RB1), have been observed in a variety of hematological malignancies including myelodysplastic syndrome (MDS). We describe here a novel unbalanced translocation der(13)t(7;13)(p13;q14) involving 13q14 in a patient with MDS. A 66-year-old woman was diagnosed as having MDS, refractory anemia with excess of blasts (RAEB-1) because of 7.4% blasts and trilineage dysplasia in the bone marrow cells. G-banding and spectral karyotyping analyses showed complex karyotypes as follows: 46,XX,der(6)t(6;7)(q11;?),der(7)del(7)(?p13)t(6;7)(q?;q11)t(6;13)(q?;q?),der(13)t 7;13)(p13;q14). Fluorescence in situ hybridization (FISH) analyses demonstrated that one allele of the RB1 gene and the microsatellite locus D13S319, located at 13q14 and telomeric to the RB1 gene, was deleted. Considering other reported cases, our results indicate that submicroscopic deletions accompanying 13q14 translocations are recurrent cytogenetic aberrations in MDS. The RB1 gene or another tumor suppressor gene in the vicinity of D13S319, or both, may be involved in the pathogenesis of MDS with 13q14 translocations by monoallelic deletion. PMID- 16213366 TI - Constitutional partial 1q trisomy mosaicism and Wilms tumor. AB - We report on a female patient with severe-profound mental retardation, multiple congenital anomalies, as well as a history of mosaicism for partial 1q trisomy in the amniotic fluid and a previous Wilms tumor specimen. Peripheral blood and fibroblasts were studied and did not demonstrate the mosaicism initially detected for 1q. Array comparative genomic hybridization yielded negative results. Additional cytogenetic studies helped clarify the previous findings and revealed evidence of partial 1q trisomy mosaicism in normal kidney tissue and in a kidney lesion. GTG-banded results showing low-percentage mosaicism for the structural rearrangement der(1)t(1;1)(p36.1;q23) in both tissues were corroborated by fluorescence in situ hybridization studies. We hypothesize that the partial 1q trisomy predisposed the target tissue (in this case kidney) to neoplasia. This study provides further support for the hypothesis that certain constitutional chromosomal abnormalities can predispose to cancer. As detection of a low percentage mosaicism may be hampered by the limits imposed by currently available technology and the constraint of a finite sample size, extra vigilance in monitoring other somatic tissues will be needed throughout the patient's lifetime. Anticipatory clinical guidance and prognostication are meaningful only if given accurate cytogenetic diagnoses. To the best of our knowledge, this is the first reported case of Wilms tumor associated with constitutional partial 1q trisomy, either in pure or mosaic form, with the particular 1q23 breakpoint in conjunction with a break on 1p36.1. PMID- 16213367 TI - Constitutional trisomy 8 mosaicism with myelodysplastic syndrome complicated by intestinal Behcet disease and antithrombin III deficiency. AB - Trisomy 8 is the most common acquired chromosomal abnormality associated with myeloid malignancy. As a constitutional trisomy 8 mosaicism (T8M), it exhibits an extremely variable phenotype. In addition, Behcet disease (BD) has been reported as an unusual complication of myelodysplastic syndrome (MDS). To our knowledge, 12 case reports of various hematologic malignancies in patients with T8M and 18 case reports of MDS with acquired trisomy 8 complicated by BD have been published to date. We report a case of constitutional T8M with MDS complicated by intestinal BD and antithrombin III deficiency. PMID- 16213369 TI - Cryptic MLL-AF10 fusion caused by insertion of duplicated 5' part of MLL into 10p12 in acute leukemia: a case report. AB - Chromosomal translocations involving the mixed lineage leukemia gene (MLL) located at 11q23 belong to common chromosomal abnormalities in both acute lymphoblastic (ALL) and acute myeloid leukemias (AML). It has been suggested that the mechanism of MLL leukemogenesis might be a result of a gain-of-function effect of the MLL fusion gene and simultaneous loss of function of one of the MLL alleles (haploinsufficiency). One of the recurrent translocations in AML-M5 involves chromosomal locus 10p12 and results in the MLL-AF10 fusion gene. Several mechanisms leading to MLL-AF10 fusion have been reported, and they have involved rearrangement of the 11q23 region. We present a detailed structural analysis of an AML case with an extra copy of the 5' part of MLL region and its insertion into the short arm of chromosome 10, resulting in an MLL-AF10 fusion without rearrangement of the MLL alleles on both chromosomes 11. Our observation supports a role for a simple MLL gain-of-function in leukemogenesis. PMID- 16213368 TI - The presence of TEL/AML1 rearrangement and cryptic deletion of the TEL gene in adult acute lymphoblastic leukemia (ALL). AB - TEL/AML1 (also known as ETV6/RUNX1) rearrangement is the most frequent genetic change in childhood B-acute lymphoblastic leukemia (ALL) and is associated with a favorable prognosis. Its presence in adult ALL, however, has been questionable, and any association between TEL/AML1 rearrangement and clinical prognosis is unknown. To reveal the presence and incidence of the TEL/AML1 rearrangement in adult ALL, we applied fluorescence in situ hybridization (FISH). We conducted extra-signal, dual-color fluorescence in situ hybridization (ES-FISH) for TEL/AML1 rearrangement on bone marrow cells from 74 adult ALL patients and analyzed the survival time. We demonstrated the TEL/AML1 rearrangement in 2 patients (2.7%) among 74 patients with ALL. Of 74 adult ALL patients, 3 (4.0%) showed deletion of the TEL gene without TEL/AML1 rearrangement. The mean survival time of patients with TEL/AML1+ALL and patients with cryptic TEL deletion was 33 and 5 months, respectively, whereas the mean survival time of 53 TEL/AML1-ALL patients was 16.7 months. TEL/AML1 rearrangement is not unique in childhood ALL, and cryptic TEL deletion without TEL/AML1 rearrangement was more frequent than the TEL/AML1 rearrangement in adult ALL. We recommend TEL/AML1 FISH in adult ALL patients because conventional cytogenetic studies so far have yielded uninformative results. PMID- 16213370 TI - IgM myeloma with t(4;14)(p16;q32). PMID- 16213373 TI - Risk factors for endophthalmitis following cataract surgery: a retrospective case control study. AB - To gain further knowledge of possible risk factors for the development of postoperative endophthalmitis (POE) following cataract surgery, a single-centre retrospective case-control study was conducted for 1994-2000. In total, 46292 cataract procedures were performed during the study period. Sixty cases of POE were noted and 240 control cases were selected at random. Parameters pertaining to patient history and to peri-operative technique and complications were analysed. The relative risk of POE was calculated using univariate analyses and multi-variate forward stepwise logistic regression. In the logistic regression analyses, three statistically significant parameters were found. The use of intracameral prophylaxis with cefuroxime as opposed to topical treatment alone, and performing phaco-emulsification instead of extra- or intracapsular cataract extraction appeared to be protective against POE. Silicone intra-ocular lenses carried a higher risk than heparin surface modified poly (methyl methacrylate) implants. In summary, the important finding of this study was the protective effect against POE of intracameral prophylaxis with cefuroxime compared with topical anti-infectives alone. PMID- 16213372 TI - An outbreak of Pantoea spp. in a neonatal intensive care unit secondary to contaminated parenteral nutrition. AB - Contaminated parenteral nutrition (PN) is an important source of infection in neonates. Many organisms have been reported to cause contamination that results in outbreaks in intensive care units. The objective of this study was to investigate an outbreak caused by Pantoea spp., which contaminates PN, in a neonatal intensive care unit (NICU). This was a descriptive study of an outbreak of sepsis in an NICU of a tertiary teaching hospital in Malaysia. Pantoea spp. infection was detected in eight patients over a three-day period from 24 to 27 January 2004 following the administration of PN. Seven of the eight patients died due to the infection. Extensive environmental samplings for culture were performed. PN solution from the NICU and the pharmacy were also cultured during the outbreak period. Pantoea spp. was isolated from blood cultures of all infected patients, and the unused PN from the pharmacy and the NICU. All the strains of Pantoea spp. had a similar antibiotic susceptibility pattern and biochemical reaction. From the results, we concluded that PN was the source of the outbreak and the contamination may have occurred during its preparation in the pharmacy. A thorough investigation has been carried out and, where possible, corrective measures have been taken to avoid similar outbreaks in the future. PMID- 16213376 TI - Orthobiologics: a survey of materials and techniques. AB - The modern study of the regenerative capabilities of skeletal tissues began in 1964, when Marshall Urist discovered that bone devoid of inorganic content and dried into a powder could be implanted into the muscle of a rabbit with resultant ectopic bone formation. Use of this technology is now commonplace, and a plethora of orthobiologic materials are available to the foot and ankle surgeon. The science has even become advanced to the point that custom bone replacements can be fabricated in vitro. This points toward an exciting future for the use of this technology by foot and ankle surgeons. PMID- 16213377 TI - Bioscaffolds in tissue engineering: a rationale for use in the reconstruction of musculoskeletal soft tissues. AB - Bioscaffolds derived from animal tissues can be an appealing substrate to induce the formation of functional tissue (histogenesis) within the context of tissue engineering. Bioscaffolds obtained from the extracellular matrix not only contain collagen, which can provide mechanical support, but also include the required biologically active molecules that provide a stimulus for active tissue remodeling. Manufacturing, processing, and the tissue source of the biological scaffold affect the biologic outcome and are important in predicting the clinical results. This article discusses the merits and limitations of using bioscaffolds in soft tissue engineering. PMID- 16213378 TI - The use of acellular biologic tissue patches in foot and ankle surgery. AB - A new area of orthobiologics is biologic patches. These are acellular tissue grafts providing a regenerative tissue matrix. The biologic patch serves two functions: they reinforce the soft tissues during the healing process, and they provide a scaffold of collagen for cellular ingrowth. They are excellent alternatives to autografts, allografts, and synthetic materials. PMID- 16213379 TI - Tendon repair and healing. AB - Tendon connects muscle to bone and functions to transmit muscular forces across joints to stabilize or move those joints. Tendons in the foot and ankle are subject to enormous loads and consequently make up a substantial portion of the body's tendon injuries. Understanding the mechanisms of these injuries requires an understanding of the relative rates of muscle, tendon, osteotendinous junction, and myotendinous junction adaptation. This article provides the practitioner with an overview of tendon anatomy, physiology, healing, and repair and correlates didactic and clinical aspects so that practitioners can better treat patients and get them back to normal functioning as quickly and as close to anatomic and physiologic capabilities as possible. PMID- 16213380 TI - The utilization of autologous growth factors for the facilitation of fusion in complex neuropathic fractures in the diabetic population. AB - A review of current knowledge of autologous growth factors as used in foot and ankle surgery is presented. This knowledge is clinically correlated with 50 Charcot's foot reconstruction patients who had diabetes and who were randomized to a platelet-rich plasma (PRP) concentration system (Symphony, DePuy, Warsaw, Indiana) or a hollow-fiber hemoconcentration system (Interpore Cross AGF, Interpore Cross, Irvine, California) trial. Although the literature supports the notion that Symphony produces a higher yield of intact platelets more consistently, clinically, a statistically significantly higher number of patients treated with Interpore Cross AGF went on to solid fusion. The findings may indicate that one type of PRP may be indicated for a particular clinical circumstance based on the patient's medical history and resultant local wound environment. PMID- 16213381 TI - Viscosupplementation (hyaluronans) in the treatment of ankle osteoarthritis. AB - This pilot study was undertaken to gather preliminary data on the efficacy and safety of viscosupplementation with sodium hyaluronate versus phosphate-buffered saline control for pain associated with osteoarthritis (OA) of the ankle. Results suggest that five weekly intra-articular injections of sodium hyaluronate in patients who have OA of the ankle are well tolerated, can provide sustained relief of pain, and improve ankle function. These findings are consistent with previously published studies using intra-articular injections of sodium hyaluronate in other articular joints but require confirmation in a large, randomized, saline-controlled study. If confirmed, these findings would provide a valuable nonoperative treatment option for patients who have OA of the ankle. PMID- 16213382 TI - Demineralized bone matrix: basic science and clinical applications. AB - Bone grafting with demineralized bone matrix (DBM) is useful in reconstructive surgery to ultimately provide anatomic alignment, restore function, or augment/change the biomechanics of the foot and ankle. DBM should be used in conjunction with a transplant or implant displaying mechanical strength. DBM can augment cortical grafts used for bridging gaps or defects and lengthening procedures by increasing the connectivity of the structural graft with the host bone. Another useful application is providing a biologic boost to patients who have less-than-ideal physiology. Because DBM has higher concentrations of available bone morphogenic proteins, it can aid in the incorporation of other grafts. Other uses include delayed unions, nonunions, packing joints for arthrodesis, filling resected cysts, and filling gaps of debrided infected bone. PMID- 16213383 TI - The bone morphogenic protein. AB - The molecular and cellular process to promote bone formation has been examined extensively in recent years in an attempt to minimize delayed unions and nonunions. Bone morphogenetic proteins (BMPs) have been determined to play an intricate role in the bone formation cascade. Over 14 BMPs have been isolated and more are being discovered as investigation progresses. BMP-2 and BMP-7 are being produced commercially for clinical use with specific indications. Continuing research is investigating the optimal carrier that will give the best results. This article reviews the most current information regarding BMPs. PMID- 16213384 TI - Bone graft substitutes: osteobiologics. AB - Future devolvement of osteobiologic materials will no doubt replace materials currently being used. As techniques to improve biointegration and manipulation of the healing environment proceed, future graft substitutes may exceed even autogenous bone in their reliability. The primary role of bone graft use in foot and ankle surgery has been to fill traumatic defects and benign tumors or to augment arthrodesis techniques. This article highlights the osteobiologics in use today, including calcium-based ceramics, hydroxyapatite, calcium phosphates and calcium composite materials, collagen composite materials, calcium sulfate, calcium cements, and allograft. PMID- 16213387 TI - Pediatric ophthalmology in South Africa. PMID- 16213385 TI - The future of bone healing. AB - With more than 5.5 million fractures and 1 million bone repair surgeries annually, bone graft plays a significant role in aiding fracture repair. In the United States alone, surgeons perform an estimated 500,000 to 600,000 bone grafting procedures annually. With the advent of possibilities from bone substitutes, growth factors, and stem cell research, the potential for enhancing bone healing is vast. This article attempts to survey current trends and to highlight upcoming techniques in the future of bone healing. PMID- 16213388 TI - Ophthalmologic findings in the Cornelia de Lange Syndrome. AB - BACKGROUND: Cornelia de Lange Syndrome (CdLS) is a disorder caused in many patients by a mutation in the NIPBL gene with a dominant pattern of inheritance characterized by mental retardation, prenatal and postnatal growth retardation, upper-limb abnormalities, and characteristic facies. Few data exist concerning the ophthalmic findings in this syndrome. METHODS: One hundred twenty individuals with CdLS underwent ophthalmic examination to ascertain the relative frequencies of oculofacial and ophthalmic abnormalities. RESULTS: We confirmed the frequent findings of synophrys (99%), long lashes (99%), hypertrichosis of the brows (96%), ptosis (44%), epiphora (22%), nasolacrimal duct obstruction (16%), blepharitis (25%), and myopia (58%). In addition, we found peripapillary pigment (83%), and microcornea (21%), which have infrequently been mentioned in the literature. CONCLUSION: Patients with CdLS can have multiple eye problems. Many of these problems can be readily treated, including myopia, blepharitis, nasolacrimal duct obstruction, and ptosis. Early examination is recommended for all children known or suspected to have CdLS. PMID- 16213389 TI - Vertical rectus muscle transposition for bilateral Duane syndrome. AB - BACKGROUND: Augmented transposition of the superior and inferior rectus muscles to the lateral rectus muscle is effective surgical treatment for esotropia in unilateral Duane syndrome. Medial rectus muscle recession in bilateral Duane syndrome may increase the risk of consecutive exotropia and cause limitation to adduction postoperatively. Vertical rectus muscle transposition may be useful in bilateral Duane syndrome with esotropia. METHODS: We undertook a retrospective review of 11 patients with bilateral Duane syndrome and esotropia in primary position. All patients had vertical rectus muscle transpositions. Six patients had unilateral vertical rectus transpositions (2 eyes with and 4 without suture augmentation). Twelve eyes from 7 children (2 unilateral and 5 bilateral) had transpositions augmented with posterior fixation sutures. Posterior fixation suture were added to large deviations in patients without prior medial rectus recessions. RESULTS: The preoperative esotropia at distance was 22.8 +/- 6.3 prism diopters (PD). It reduced to 2.0 +/- 6.7 PD postoperatively. (P < 0.001) Esotropia at near changed from 21.0 +/- 5.8 PD preoperatively to 1.2 +/- 8.1 PD postoperatively. (P < 0.001) One patient with a 10-degree face turn had complete resolution postoperatively. One patient had a small undercorrection and developed a vertical deviation requiring additional surgery. All patients had improvement in abduction. Nine of 11 patients did not develop any limitation to adduction. One patient developed a -1 adduction deficit 5 years later. Three patients achieved fusion with a mean stereovision of 67 seconds of arc (range, 80-40 seconds.). Follow-up averaged 22.2 months (range, 1-100 months). CONCLUSION: Vertical rectus muscle transposition in patients with bilateral Duane syndrome and esotropia is an effective procedure to improve ocular alignment and motility while preserving adduction. PMID- 16213390 TI - Strabismus in senile cataract patients. AB - PURPOSE: We sought to determine the prevalence of strabismus among patients with senile cataract. METHODS: Forty-seven consecutive patients who were scheduled for cataract surgery underwent complete eye examinations, including ocular alignment evaluation. Twenty-four normal adults of the same age range underwent also complete eye examinations and were used as a control group. All patients of the cataract group underwent unilateral cataract surgery. The angle of strabismus was measured by the prism and cover test or prism and corneal reflex test. RESULTS: Mean preoperative deviation of the study group was 17.8 +/- 9.7 prism diopters (PD) of exophoria/tropia. In this group, 70.2% had exophoria and 27.7% had exotropia. In the control group exophoria was found in 75% of the persons whereas none of them had any heterotropia. Mean deviation in the control group was 4.7 +/ 5.1 PD of exophoria. Postoperatively, the angle of exophoria/tropia improved to 12.8 +/- 8.5 PD, which was different from the preoperative measurements (P < 0.01). In this group 78.8% had exophoria and 19.1% had exotropia. CONCLUSIONS: Patients with cataract have greater tendency to develop exotropia or exophoria than noncataract persons of the same age. Cataract surgery improves the heterophoric status of these patients. PMID- 16213392 TI - The effect of bilateral horizontal rectus recession on visual acuity, ocular deviation or head posture in patients with nystagmus. AB - PURPOSE: This work was undertaken to evaluate the effect of bilateral horizontal rectus recession on visual acuity, ocular deviation and head posture in patients with nystagmus. METHODS: Twenty patients underwent recession of 4 horizontal rectus muscles to dampen the nystagmus. At least 2 muscles were recessed posterior to the equator. RESULTS: Monocular and binocular visual acuity improved from 1 to 3 Snellen lines after the procedure in 13 patients (76.5%). Visual changes in log MAR notations were statistically significant with Wilcoxon analysis in each and both eyes. Preoperative binocular visual acuity was 0.73 +/- 0.26 log MAR that reached to 0.62 +/- 0.32 log MAR after surgery (P = 0.02). The greatest improvement was observed in patients with congenital motor nystagmus. Ten patients had horizontal strabismus in addition to nystagmus before the surgery. One-millimeter additional recession of both medial rectus muscles caused an average reduction of 6 PD in esotropia. Corresponding figures for similar additional recessions of the lateral rectus muscles was 11 PD for correction of exotropia. Abnormal head posture decreased in all the cases and improved completely in most of them. CONCLUSIONS: Large horizontal rectus recession can improve visual acuity and decrease nystagmus in sensory and motor types. By revision in surgical planning, strabismus and abnormal head posture can also be corrected. PMID- 16213391 TI - Ocular complications after organ and bone marrow transplantation in children. AB - BACKGROUND: Organ and bone marrow transplantation commonly are performed in children. Ocular complications usually are described as secondary to post transplantation medications. The complication rate is unknown. METHODS: A retrospective chart review was performed of 93 children who were younger than 18 years of age and had transplantation surgery from 1989 to 2004. The rate and type of ocular complications, including those requiring ocular surgery, were analyzed. Medications and visual loss associated with adverse effects also were studied. RESULTS: Of the 93 patients, 74 patients met the entry criteria. Sixty-one patients had at least 1 year of follow-up, and the longest follow-up duration was 14 years. The 1-year post-transplantation complication rate was 16.0% (95% confidence interval 6.8-24.4%). Adverse effects included cytomegalovirus (CMV) retinitis, cataract, graft-versus-host disease, lymphoproliferative disorder, persistent strabismus, and transient visual loss. Four patients underwent eye surgery, including lensectomy for cataract, tarsorrhaphy for corneal ulcer, and iris biopsy. They had surgery 0.9 to 4.7 years after transplantation. Most patients were taking prednisone and cyclosporine when their complication was diagnosed. One patient's visual acuity deteriorated to no light perception in one eye and 20/250 in the other eye secondary to CMV retinitis. Most patients had a final visual acuity > or =20/40. CONCLUSION: Transplantation surgery in children produces a significant risk of ocular impairment. The 1-year complication rate was 16.0%. Eye surgery may be required within the first few years after transplantation. Although most patients maintained a final visual acuity of 20/40 or better, one patient became bilaterally legally blind. PMID- 16213393 TI - High-resolution magnetic resonance imaging demonstrates varied anatomic abnormalities in Brown syndrome. AB - INTRODUCTION: Although Brown syndrome classically is considered to be limited to the SO tendon sheath and trochlea, it does not always respond to SO surgery. We investigated mechanisms of Brown syndrome by magnetic resonance imaging (MRI). METHODS: Three patients with congenital and 8 with acquired Brown syndrome were compared with matched normal subjects under a prospective protocol of high resolution, multipositional orbital MRI using surface coils. Muscle size and contractility were determined using digital image analysis. RESULTS: Five of 8 patients with acquired Brown syndrome had a history of trauma or surgery and demonstrated extensive scarring, avulsion, or fracture of the trochlea. One of the 8 had a cyst in the SO tendon. One congenital and one acquired case demonstrated inferior displacement of the lateral rectus (LR) pulley in adduction, with a normal SO tendon-trochlear complex. Such cases of Brown syndrome responded to surgical stabilization of the LR pulley. Two congenital cases had clinical findings of ipsilateral SO palsy confirmed on MRI by atrophy or absence of the SO belly. In congenital absence of the SO belly, the anterior tendon was present but terminated directly on the trochlea. CONCLUSION: High resolution MRI demonstrates a variety of abnormalities in patients presenting with Brown syndrome, including atrophy or absence of the SO belly. Management in Brown syndrome should be tailored to the pathophysiology of the individual patient. PMID- 16213394 TI - Visual outcome and success of amblyopia treatment in unilateral small posterior lens opacities and lenticonus initially treated nonsurgically. AB - PURPOSE: We sought to assess the success of amblyopia treatment in patients with small posterior lens opacities as well as the factors associated with a good visual outcome. METHODS: This was a retrospective study of patients with posterior lens opacities that initially were thought to be too small in size to warrant cataract surgery. The following variables were examined: cataract type, location, diameter, persistent hyaloid vessel, anisometropia, strabismus, and age of detection. Success of treatment of amblyopia was defined as improvement by at least 0.3 logMAR units. Good visual outcome was defined as 20/40 or better. Amblyopia was treated by glasses, patching, and/or atropine. Patients who failed with conservative treatment or had an increase in cataract size underwent surgery. RESULTS: Forty-eight (91%) of 53 eyes were amblyopic. Thirty amblyopic eyes had pre- and post-treatment Snellen acuities. Twenty (67%) had their visual acuity (VA) improved by 0.3 logMAR units or greater. None of the measured variables were associated with successful amblyopia treatment. Twenty-five (49%) of 51 patients had a final VA of 20/40 or better. The only variable associated with good visual outcome was cataract type: 18 of 25 (72%) posterior subcapsular cataract and 6 of 23 (32%) posterior lenticonus eyes achieved VA of 20/40 or better (P = 0.008). Six patients who went on to have cataract surgery experienced a larger improvement in BCVA (4.50 logMar units +/- 2.52 lines) compared with patients treated without cataract surgery (2.36 logMar units +/- 3.11 lines). DISCUSSION: Amblyopia treatment was successful in most cases. A small group of patients who underwent cataract surgery experienced a greater VA improvement; however, it was not statistically significant. Further studies are needed to determine which patients would benefit from cataract surgery. PMID- 16213395 TI - The management of strabismus in adults--III. The effects on disability. AB - INTRODUCTION: This is the third article in a series on the various facets of the management of strabismus in adults. Here, we give a broad overview of the types and severity of disability and provide initial validation of an instrument (questionnaire) to assess these disability aspects. METHODS: After undergoing strabismus surgery, 101 patients from 6 centers completed a 6-item questionnaire in which they rated both the before-surgery and after-surgery severity of problems associated with their strabismus, ranging from specific health, daily functioning, social interaction, concerns about the future, and self-image to job related difficulties. RESULTS: The before-surgery outcomes showed significant variation across the 6 types of problems (P < 0.001), with "specific health" and "daily tasks" yielding the highest problem rating. Patients with diplopia reported more severe problems with "daily tasks" (P = 0.004) and "concerns about the future" (P = 0.026) than patients without diplopia. Overall, all problem ratings declined after surgery (P < 0.001), but patients who were not successfully aligned were left with higher problem ratings on "specific health" (P = 0.005), "daily tasks" (P = 0.003), and "social interaction" (P = 0.024). CONCLUSIONS: The results indicate a wide range of disability aspects in adult patients with strabismus, with moderate differences between patients with or without diplopia. Improvements in disability after surgery, as reflected by these ratings, should be taken into account when assessing the health value of adult strabismus management. PMID- 16213396 TI - Intermittent exotropia, overcorrecting minus lenses, and the Newcastle scoring system. AB - PURPOSE: We sought to determine whether the Newcastle Control Score (NCS) could be used to indicate a successful outcome in patients with intermittent exotropia that were treated with minus lenses. METHODS: We studied patients with an intermittent exotropia who were prescribed minus lens therapy in an effort to manage their strabismus conservatively. The NCS, which quantitatively estimates the control of an intermittent exotropia, was applied before treatment and 4 months after treatment. The results of minus lens therapy with a pretreatment NCS of > or =5 (group 1) were compared with those with a NCS of < or =4 (group 2). RESULTS: There were 24 patients (13 girls, 11 boys) treated with minus lenses. The mean age of the patients was 6.8 +/- 3.8 years (range, 2-17 years; median, 5). The mean pretreatment distance angle was 28.5 +/- 10 prism diopters (range, 6 45; median, 30) and the mean post-treatment distance angle was 18.3 +/- 8.9 prism diopters (range, 0-35; median, 18) P = 0.001. Using the NCS minus lenses significantly (P = 0.041) improved control of exotropia. In group 2 (n = 16) 75% had improved scores post-treatment compared with 62.5% in group 1 (n = 8), P = 0.68. CONCLUSIONS: These data suggest that the NCS is a useful method to indicate the success in the control of intermittent exotropia with conservative treatment with minus lens therapy. PMID- 16213397 TI - Balloon dilation of the nasolacrimal duct. AB - PURPOSE: Our purpose is to report our experience with balloon catheter dilation of the nasolacrimal duct after failed probing for congenital nasolacrimal duct obstruction. METHODS: We undertook a retrospective chart review. Patients who were enrolled had symptoms of persistent nasolacrimal duct obstruction after simple probing and irrigation. Balloon nasolacrimal ductoplasty was performed using the LacriCATH system (Quest Medical, Inc. An Atrion Company, Allen, TX). Outcomes were determined by postoperative examination and telephone follow-up. RESULTS: Thirty-nine lacrimal systems of 26 patients (age range, 10 months to 84 months) were treated. Of the obstructed ducts treated, 82% (32/39) showed resolution of epiphora. There were 9 children older than the age of 2, with 15 nasolacrimal systems studied. These had 11 of 15 successes for a success rate of 73%. There were 17 children between the ages of 10 months and 2 years with 24 nasolacrimal systems studied. These had 21 of 24 successes for a success rate of approximately 88%. The difference was not statistically significant (P = 0.28). CONCLUSION: Balloon dilation of the nasolacrimal duct is an alternative to silicone intubation in the treatment of congenital nasolacrimal duct obstruction after failed simple probing. Although our success rate was slightly lower than some published reports of nasolacrimal system intubation, this simple and atraumatic procedure was successful in 82% of cases for this specific situation. PMID- 16213398 TI - Contrast sensitivity in amblyopia: the fellow eye of untreated and successfully treated amblyopes. AB - PURPOSE: We sought to investigate contrast sensitivity on the fellow eyes of amblyopic and successfully treated amblyopic subjects. METHODS: Contrast sensitivity was tested monocularly on both eyes of 48 amblyopic patients (mean age, 11.51 years) and of 22 successfully treated amblyopic subjects (visual acuity 20/20 in each eye; mean age, 11.22 years). Inclusion criteria were visual acuity in the amblyopic eye 20/40 or better (mild amblyopia) and 20/20 or better in the fellow eye, steady fixation, no signs of congenital, latent or manifest/latent nystagmus on clinical examination. Twenty normal subjects (20 eyes) were used as age-matched controls. RESULTS: Contrast sensitivity functions from the fellow eye of the 48 amblyopic patients, even those who had never been treated with occlusion therapy before, were significantly decreased (P < 0.001) compared with control subjects. Both the previously amblyopic and the fellow eye of the 22 "cured" amblyopic subjects demonstrated significantly lower values (P < 0.001) compared with control patients. CONCLUSIONS: We suggest that the nonamblyopic, "normal" eye of amblyopic patients behaves abnormally when evaluated for contrast sensitivity functions. Neither the previously amblyopic nor the fellow eyes of successfully treated subjects were comparable with controls. Occlusion therapy may not be implicated for depressed contrast sensitivity of the fellow eye in amblyopia. The assessment of contrast sensitivity can provide important information on the visual function and the influence of occlusion therapy in amblyopia. PMID- 16213399 TI - Are norms based on the original Teller Acuity Cards appropriate for use with the new Teller Acuity Cards II? AB - PURPOSE: We sought to compare visual acuity results obtained with the original Teller Acuity Cards (TAC), which are no longer commercially available, and the modified, laminated Teller Acuity Cards II (TAC II), which recently became commercially available. METHODS: Sixty children were tested, 20 at each of 3 ages: 3.5 months, 11 months, and 30 months. Each subject's binocular grating acuity was measured once with the TAC and once with the TAC II, with the order of testing counterbalanced across subjects. Testers were aware that acuity cards were arranged in ascending order of spatial frequency, but they were masked to the absolute spatial frequencies of the gratings in the subset of cards used. Testers were also masked to acuity results until testing of the child was completed. RESULTS: Repeated-measures analysis of variance with age as a between subjects variable and card type as a within-subjects variable showed a significant effect of age (P < 0.001) and a significant effect of card type (P < 0.001), but no interaction between age and card type. Post hoc comparisons (with Bonferroni correction) showed that mean acuity score was significantly better with TAC than with TAC II at 3.5 months (0.2 octave, P < 0.05), 11 months (0.4 octave, P < 0.01), and 30 months (0.7 octave, P < 0.001). CONCLUSIONS: These results suggest that normative grating acuity data obtained with the original Teller Acuity Cards need to be adjusted toward lower acuity values by approximately 0.5 octave to be appropriate for use with the new Teller Acuity Cards II. PMID- 16213400 TI - Could personality traits modify the response to uncorrected high hyperopia? AB - PURPOSE: We aimed to measure temperament and investigate personality in children with high hyperopia considering that these could modify the individual response to uncorrected high hyperopia. METHODS: Fifteen children (age range, 5 to 12 years) with orthotropia and ametropic amblyopia in the presence of uncorrected high hyperopia were identified (group 1). Among the children with refractive accommodative esotropia, 15 children (age range, 5 to 12 years) were enrolled to form group 2. We measured the temperament by using the Children's Behavior Questionnaire (CBQ) Short Form. The Children's Apperception Test (CAT-H) was also administered to all subjects. RESULTS: Regarding the temperament scales measured by CBQ, themes of discomfort, fear, and shyness were more dominant in group 2; high-intensity pleasure and smiling-laughter were dominant in group 1. However the difference was statistically significant only for fear scale (P = 0.045). CAT H results revealed that aggression toward the parents was the most commonly encountered behavioral pattern in both groups. The children in group 1 were more likely to express passive-aggressive behavioral pattern. Obstinacy and anal period characteristics dominate in the children in group 2. Themes of narcissistic injury was more frequently expressed by these children. CONCLUSION: The temperament scales and personality traits could play a role in the modification of the individual response to uncorrected high hyperopia. This finding deserves more research in a larger study group. PMID- 16213401 TI - The effect of occlusive patching on visually-directed tasks. AB - PURPOSE: Occlusive patching is used to treat a variety of ocular conditions. A paucity of literature exists regarding the effects of occlusion on functional tasks and driving performance. We investigated the immediate effects of occlusive patching on a variety of visually dependent tasks as well as driving performance on a simulator. METHODS: Thirty normally sighted subjects were examined in a prospective randomized manner. All subjects underwent a complete medical history and eye examination. All subjects performed a variety of near tasks and drove a driving simulator with and without patching. The tasks included traditional clinical depth tests, tests of visual memory, scanning, and tracking, and gross and fine-motor abilities. All subjects filled out a questionnaire relating to their perception of the effects of patching on functional tasks and driving ability. RESULTS: Visual memory, scanning, tracking, and perceptual constancy were unaffected by patching. However, there were significant differences between the patched and unpatched conditions for four of five of the three-dimensional visual coordination tasks. Eight of 19 (42%) of the fine-motor tasks and 4 of 9 gross-motor tasks (44%) showed significant differences between the patched and unpatched conditions. A greater frequency of out of lane events and more abrupt braking profiles were seen when subjects drove monocularly rather than binocularly. CONCLUSIONS: Sudden occlusion immediately changes perceptual ability by decreasing visual field and eliminating stereoacuity. This has important implications for the performance of everyday tasks. PMID- 16213402 TI - Traumatic cataract in a 7-year-old boy caused by low-velocity impact with a soft core baseball. PMID- 16213403 TI - Extensive persistent pupillary membranes: conservative management. AB - Extensive persistent pupillary membranes (PPMS) may appear to occlude the pupil so completely as to preclude useful vision. They often motivate aggressive interventions. We present a 6-year-old girl with bilateral extensive densely pigmented pupillary membranes in whom good vision was obtained without pharmacologic, laser, or surgical intervention. There was associated ametropic amblyopia and exotropia in the patient, which were effectively treated with spectacles and occlusion therapy, with the patient responding conventionally. In view of our findings, we recommend a trial of conservative management before surgical intervention even in cases of dense PPMs. PMID- 16213404 TI - Secondary artisan phakic intraocular lens for correction of progressive high myopia in a pseudophakic child. AB - An 8-week-old infant underwent unilateral cataract extraction and posterior chamber intraocular lens implantation for total cataract in the left eye. After surgery, a residual progressive myopic error was observed, ranging from -4.5 diopters (D) 6 months after the operation to -14.0 D at the age of 3 years. Because of parental noncompliance for contact lens and spectacles use, secondary implantation of Artisan phakic IOL of -14.0 D power was performed. No intra- or postoperative complications were observed. Nine months after this second operation, myopia diminished to -3.25 D. PMID- 16213405 TI - Severe ocular surface disease and glaucoma in a newborn with aniridia. PMID- 16213406 TI - Type 3 Gaucher's disease in a three-year-old child: saccadic eye movements analysis. AB - Gaucher's disease (GD) is an autosomal-recessive disorder that leads to the storage of sphingolipid material (glucocerebroside) in different peripheral tissues and sometimes in the central nervous system. Among its three existing forms, the most frequent non-neurological form (type 1: GD1) is treatable with appropriate amounts of exogenous enzyme-replacement therapy (ERT), whereas in the type 3 form (GD3), progression of the neurological involvement may be slowed down or halted by much higher doses of ERT than those used in GD1 because of the inability of ERT to cross the blood-brain barrier. PMID- 16213407 TI - Electronic lab notebooks - do they work in reality? PMID- 16213408 TI - Rational design of peptide drugs: avoiding aggregation. PMID- 16213409 TI - Endocannabinoids mediate age-old pain suppression. PMID- 16213410 TI - Battle against malaria could involve anti-HIV drugs. PMID- 16213411 TI - Yike Guo and Jonathan Sheldon of InforSense discuss the impact of workflow technology on drug discovery. Interview by Christopher Watson. PMID- 16213412 TI - Florida focused on growing life sciences cluster. PMID- 16213413 TI - Evolution of biomarkers: drug discovery to personalized medicine. PMID- 16213414 TI - Keynote review: the adipocyte as a drug discovery target. AB - The adipocyte has pleiotropic functions beyond the storage of energy in times of nutrient abundance. Considerable efforts in adipocyte biology within the past ten years have emphasized the important role of adipose tissue in processes as diverse as energy metabolism, inflammation and cancer. Adipocytes are able to communicate with the brain and peripheral tissues implementing metabolic signals such as satiety, food intake and energy expenditure. Despite its huge pharmacological potential, only a small number of clinical applications interfere directly with adipocyte physiology. Here, we want to highlight various areas of adipocyte physiology that have not yet been explored pharmacologically and emphasize some of the limitations associated with these pharmacotherapies. PMID- 16213415 TI - Therapeutic applications of superantibodies. AB - Superantibody technology represents a method to enhance the potency and utility of monoclonal antibodies. The blueprint for superantibody technology is taken from rare naturally occurring superantibodies with unique sequence regions, conferring specific biological functions not detected on most antibodies. In superantibody technology, peptides with specific amino acid sequences are crosslinked to antibodies using affinity-site-specific chemistry. Three types of superantibodies have been engineered: dimerizing superantibodies with enhanced effector potency, superantibodies with the ability to penetrate living cells and superantibodies as vaccines with built-in molecular adjuvant. Collectively, superantibody technology generates a new class of antibodies with higher levels of therapeutic potency. PMID- 16213416 TI - BiTEs: bispecific antibody constructs with unique anti-tumor activity. AB - Bispecific T-cell engager molecules (BiTEs) constitute a class of bispecific single-chain antibodies for the polyclonal activation and redirection of cytotoxic T cells against pathogenic target cells. BiTEs combine a unique set of properties that have not yet been reported for any other kind of bispecific antibody construct, namely extraordinary potency and efficacy against target cells at low T-cell numbers without the need for T-cell co-stimulation. Here we review novel insights into the mechanism of BiTE action, which help to explain the unique features of BiTEs, as well as data from various animal models demonstrating the outstanding therapeutic potential of BiTEs for the treatment of malignant diseases. PMID- 16213417 TI - Using microarray gene signatures to elucidate mechanisms of antibiotic action and resistance. AB - Microarray analyses reveal global changes in gene expression in response to environmental changes and, thus, are well suited to providing a detailed picture of bacterial responses to antibiotic treatment. These responses are represented by patterns of gene expression, termed expression signatures, which provide insight into the mechanism of action of antibiotics as well as the general physiological responses of bacteria to antibiotic-related stresses. The complexity of such signatures is challenging the notion that antibiotics act on single targets and this is consistent with the concept that there are multiple targets coupled with common stress responses. A more detailed knowledge of how known antibiotics act should reveal new strategies for antimicrobial drug discovery. PMID- 16213418 TI - Artificial intelligence and robotics in high throughput post-genomics. AB - The shift of post-genomics towards a systems approach has offered an ever increasing role for artificial intelligence (AI) and robotics. Many disciplines (e.g. engineering, robotics, computer science) bear on the problem of automating the different stages involved in post-genomic research with a view to developing quality assured high-dimensional data. We review some of the latest contributions of AI and robotics to this end and note the limitations arising from the current independent, exploratory way in which specific solutions are being presented for specific problems without regard to how these could be eventually integrated into one comprehensible integrated intelligent system. PMID- 16213420 TI - Ruptured left cornual gestation in an unstable patient. PMID- 16213421 TI - Sigmoid penetration by an intrauterine device. PMID- 16213422 TI - Can you handle the truth (and know it when you see it)? Understanding sensitivity, specificity, predictive values, and ROC curves. AB - The interpretation of diagnostic and screening tests requires a basic knowledge of sensitivity, specificity, and predictive values. This article provides an overview of these measures of test performance, a brief description of receiver operating characteristic curves, and relevant illustrations from the medical literature. PMID- 16213423 TI - Intraoperative sigmoidoscopy in gynecologic surgery. AB - Intraoperative sigmoidoscopy is underused by the majority of practicing gynecologists and is not widely taught in obstetrics and gynecology training programs. In this report, a step-by-step approach is provided in order to perform sigmoidoscopy. Indications for use, along with various intraoperative applications, are discussed. Results from our center's experience with its use during laparoscopic treatment of adhesions, endometriosis, and associated disease of the bowel also are provided. Intraoperative sigmoidoscopy is a safe and efficacious procedure that can aid in the evaluation and treatment of pelvic pathology and facilitate identification and management of bowel injuries. It should be considered a valuable adjunct when such cases are encountered by gynecologic and pelvic surgeons. PMID- 16213424 TI - Diagnostic minihysteroscopy with vaginoscopic approach: rationale and advantages. AB - Diagnostic hysteroscopy still is considered an invasive and painful technique. In recent years, a new technique based on the employment of minihysteroscopes with fluid distention and vaginoscopic approach has been proposed in order to improve feasibility and reduce pain and side effects. Size reduction of the hysteroscope is of greatest importance for reducing pain and risk of vasovagal reaction. In conclusion, the best approach in terms of ease, reliability, acceptability, and safety to patients in whom intrauterine exploration is indicated should be simple diagnostic hysteroscopy using a small-diameter, rod lens hysteroscope with a single-flow diagnostic sheath, vaginoscopic approach, and fluid distention. In this way in about 90% of patients, a correct diagnosis and proper choice for eventual subsequent operative strategy can be obtained, without discomfort and risk. PMID- 16213425 TI - Clinical update of smooth muscle tumors of the uterus. AB - Smooth muscle tumors of the uterus represent a spectrum of diseases that range from benign leiomyoma to malignant leiomyosarcoma. The leiomyoma is the most common of these neoplasms. Clinically, it is important to fully understand the differences in clinical presentation, biologic behavior, and management for patients with benign leiomyoma, smooth muscle tumors of uncertain malignant potential, and leiomyosarcoma. The goal of this review is to present the most recent information about common smooth muscle tumors of the uterus including their etiology, histopathology, radiographic and clinical presentations, and available treatment options. PMID- 16213426 TI - Ovarian recovery after laparoscopic enucleation of ovarian cysts: insights from echographic short-term postsurgical follow-up. AB - STUDY OBJECTIVE: To evaluate damage to ovarian reserve following laparoscopic cystectomy of benign ovarian cysts. DESIGN: Prospective study (Canadian Task Force classification II-3). SETTING: Tertiary gynecologic endoscopic unit at a university-affiliated hospital. PATIENTS: Thirty-one patients who underwent excision of monolateral (n=25) or bilateral (n=6) benign ovarian cysts. INTERVENTIONS: Serial transvaginal ultrasound examinations during the first and third postsurgical menstrual cycles. The following ovarian echographic variables were evaluated: antral follicle count, ovarian volume, stromal blood flow, and side of ovulation. Two types of statistical analysis were performed: a paired analysis comparing operated and intact ovaries of the same patient and a prospective analysis comparing ecographic characteristics of the operated gonad at first and second evaluation. MEASUREMENTS AND MAIN RESULTS: Antral follicle count and stromal blood flow were not significantly affected by surgery. While ovarian volume was similar in the operated and in the contralateral intact gonad at the first ultrasound evaluation, the volume of the operated ovary was significantly reduced at the second assessment. The median (interquartile range) of the percentage of this reduction was 33% (18%-81%). This progressive reduction was confirmed by prospectively analyzing the operated ovaries. An increased probability of ovulation in the intact gonad was observed at both assessments. CONCLUSION: Laparoscopic excision of ovarian cysts is associated with damage to ovarian reserve, at least immediately after surgery. This effect does not appear to be consequent to an injury to ovarian vascularization. PMID- 16213427 TI - Oral and local anesthesia in the nonsurgical radiofrequency-energy treatment of stress urinary incontinence. AB - STUDY OBJECTIVE: To demonstrate the feasibility, safety, and patient comfort associated with nonsurgical radiofrequency-energy (RF) tissue micro-remodeling in women with stress urinary incontinence (SUI) given oral and local anesthesia. DESIGN: Prospective, open-label pilot clinical trial (Canadian Task Force classification II-2). SETTING: Department of urology in a major academic teaching hospital in Mexico City, Mexico. PATIENTS: Sixteen women with SUI and hypermobility (based on history and physical examination) with no history of previous definitive incontinence therapy. INTERVENTIONS: Nonsurgical, transurethral, outpatient RF tissue micro-remodeling with women given oral plus local anesthesia. MEASUREMENTS AND MAIN RESULTS: The women had a mean age of 49.7 years (range 30-76 years) and a mean duration of incontinence of 7.6 years (range 1-30 years). The nonsurgical RF micro-remodeling treatment, which was previously demonstrated to be of significant benefit when administered under intravenous (IV) sedation in an outpatient surgicenter setting, was successfully completed in all 16 women. Either the treating physician or the patient had the option to convert to IV sedation during the procedure if there was too much discomfort; however, this did not occur in any of the 16 patients. Thus neither the treating physician nor any patient determined that conversion to IV conscious sedation was required for treatment completion. The first six patients received an oral sedative and oral analgesic as well as a local periurethral anesthetic block with 10 mL of 2% lidocaine. The final 10 patients (63%) received only one oral sedative or analgesic and a total of 10mL lidocaine local anesthetic. Two women who received the maximum oral regimen (both oral sedation and analgesics) experienced nausea and emesis when drinking immediately after treatment, and one of these women also experienced urinary retention, which resolved after 24 hours of catheterization. Immediately before discharge, subjects classified their pain on a scale from zero ("no pain") to 10 ("terrible pain"). Mean score was 1.8, and 38% of subjects selected "zero." CONCLUSION: This pilot trial demonstrated the feasibility, safety, and patient comfort associated with performing a novel new successful technique of nonsurgical RF of the urethra for treatment of SUI, which was previously studied under IV sedation in an outpatient surgery center, on women in an office-based setting using oral plus local anesthesia. PMID- 16213428 TI - Feasibility and clinical significance of endoluminal assessment by transvaginal salpingoscopy during transvaginal hydrolaparoscopy in infertile women. AB - STUDY OBJECTIVE: To investigate the feasibility and clinical usefulness of transvaginal salpingoscopy (TS) during transvaginal hydrolaparoscopy (THL) in infertile women. DESIGN: Retrospective study (Canadian Task Force classification II-3). SETTING: Jichi Medical School, Tochigi, Japan. PATIENTS: One hundred thirty infertile women, aged 22 to 43 years, who underwent THL. INTERVENTIONS: Transvaginal salpingoscopy findings in comparison with THL findings. MEASUREMENTS AND MAIN RESULTS: Transvaginal salpingoscopy could be performed in 41.0% of the attempted tubes. In comparing rates of successful TS in relation to tubal patency, tubal morphology, and past Chlamydia trachomatis infection, patent tubes and normal-appearing tubes classified with normal morphology had a significantly higher TS success rate. However, past C. trachomatis infection did not affect the success of TS. There was a significant correlation between salpingoscopic and laparoscopic findings; however, there were no significant correlations between salpingoscopic findings and C. trachomatis antibody titers. CONCLUSIONS: These findings suggest that TS can be performed simultaneously in some infertile women undergoing THL especially those with patent tubes or with normal-appearing tubes. Further studies are required to show the usefulness of intratubal exploration in assessing tubal infertility. PMID- 16213429 TI - Development of a thinner and more flexible type of minihysteroscope with a controlled 90-degree bendable tip for vision-guided endometrium biopsy. AB - STUDY OBJECTIVE: Evaluation of the uterine cavity is limited with rigid 5-mm hysteroscopes because of the need for cervical dilatation, reduced movements inside the uterus, and no option for vision-guided biopsy. In cooperation with PolyDiagnost GmbH, Pfaffenhofen, Germany, a new type of flexible minihysteroscope with bendable tip was developed and evaluated. DESIGN: Prospective and parallel observational interindividual evaluation of flexible minihysteroscope and standard hysteroscope for diagnostic hysteroscopy (Canadian Task Force classification II-3). SETTING: Obstetrics and gynecology department of a university clinic. PATIENTS: Nine women, average age 65.0 years (range 46-89 years), with indications for diagnostic hysteroscopy. INTERVENTIONS: After defining requirements, a novel, thinner, and more flexible minihysteroscope, 18 cm long with a 2.67-mm outer diameter, was developed with straight zero-degree scope, 70-degree vision field, and 6000-pixel resolution. Two working channels, 1.2 mm and 0.55 mm, allow suction-irrigation and introduction of a 1.0-mm biopsy forceps or cytology brush. The tip of the instrument is 90-degree stageless bendable to both sides. Diagnostic hysteroscopy was performed with flexible minihysteroscope followed by standard rigid hysteroscopy to verify results. MEASUREMENTS AND MAIN RESULTS: From July 2003 through March 2004, both procedures were performed in nine patients with identical visual and histologic results. No complications occurred. No cervix-dilating instruments were necessary for introduction of the flexible minihysteroscope. Visualization of the entire uterine cavity is improved with the flexible scope because a bendable tip allows better peripheral vision (e.g., of the openings of the tubes). However, movement of the tip should be performed carefully due to potential risk of uterine perforation. CONCLUSION: This new flexible minihysteroscope is less invasive compared with standard rigid hysteroscopy, which supports performance of ambulatory hysteroscopy and makes increased movements and vision-guided biopsy inside the uterine cavity possible. PMID- 16213430 TI - The Hohl instrument for optimizing total laparoscopic hysterectomy procedures. AB - STUDY OBJECTIVE: To evaluate the feasibility of total laparoscopic hysterectomy (TLH) using the Hohl instrument in an initial cohort of patients. DESIGN: Retrospective cohort analysis (Canadian Task Force classification II-3). SETTING: Department of Obstetrics and Gynecology, Erlangen University Hospital, Erlangen, Germany. PATIENTS: Forty-four women underwent the new TLH procedure using the Hohl instrument from May 2004 through January 2005. The laparoscopic approach was used when the patient had undergone more than one previous pelvic abdominal operation and/or had a reduced vaginal capacity. The indications for hysterectomy were symptomatic leiomyoma in 25 patients and hypermenorrhea in 19 patients. INTERVENTION: Total laparoscopic hysterectomy using the Hohl instrument. MEASUREMENTS AND MAIN RESULTS: No ureteral or bladder injury occurred in any of the patients. The complication rate during surgery and in the postoperative period was zero. The mean loss of hemoglobin was 1.68+/-0.96 g/dL, the mean operating time was 108+/-21 minutes, and the mean uterine weight was 302+/-121 g. CONCLUSION: Total laparoscopic hysterectomy using the Hohl instrument simplifies the surgical procedure. The reported technique is an option comparable with laparoscopic-assisted vaginal hysterectomy and may be effective in preventing minor and major complications during TLH. PMID- 16213432 TI - Idiopathic postoperative ascites after laparoscopic salpingectomy for ectopic pregnancy. AB - Postoperative ascites is a rare and serious complication of laparoscopic procedures. Urinary tract injury, bowel injury, and lymph duct injury are the common causes; however, in some patients no definitive cause can be identified after an extensive diagnostic work-up. We present a case of postoperative ascites of unknown origin. The patient recovered with general treatments. We presume that diffuse peritoneal injury may have been caused by unidentified substances in our patient. PMID- 16213431 TI - Successful treatment of cervical stenosis with hysteroscopic canalization before embryo transfer in patients undergoing IVF: a case series. AB - The course of the transfer catheter through the cervical canal is one of the most important issues for a successful embryo transfer (ET) during in vitro fertilization (IVF) cycles. Technically difficult ETs due to cervical stenosis are associated with reduced chance of pregnancy after assisted reproductive procedures. In the current case series, we report on three patients with cervical stenosis who underwent IVF-ET cycles. These three patients, in whom ET was classified as "difficult," failed to conceive with previous ET attempts. An intervention to create a cervical tract was performed with operative hysteroscopy under general anesthesia before transcervical ET. After the hysteroscopic shaving procedure, we observed quite an improvement in access to the endometrial cavity during ET procedure. These patients had significantly easier ET procedures compared with previous attempts and achieved clinical pregnancies. Hysteroscopic revision of the cervical canal results in easier ET and improves pregnancy rates in patients with cervical stenosis and histories of difficult ET. PMID- 16213434 TI - Myomas and myomectomy. PMID- 16213441 TI - Comment on: effect of heated and humidified CO2 during gynecologic laparoscopic surgery on analgesic requirements and postoperative pain. PMID- 16213442 TI - Comment on: safe entry techniques during laparoscopy: left upper quadrant entry using the ninth intercostal space-a review of 918 procedures. PMID- 16213445 TI - The effect of the mobile phase additives on sensitivity in the analysis of peptides and proteins by high-performance liquid chromatography-electrospray mass spectrometry. AB - The study of the effect of mobile phases on sensitivity in the analysis of peptides and proteins by high-performance liquid chromatography (HPLC) electrospray mass spectrometry (ESI-MS) has been the aim of this review. Reversed phase chromatography (RPLC) is the chromatographic mode most suitable for coupling with ESI-MS since mobile phases containing organic modifiers are used. The analysis of proteins and peptides by RPLC mostly involves the use of trifluoroacetic acid (TFA) as an ion-pairing agent despite its being a strong suppressor of the MS signal. Different studies reporting the effects of using other ion-pairing agents (other perfluorinated acids, acetic acid, formic acid, etc.) and buffers (ammonium acetate, ammonium formate, ammonium bicarbonate, morpholine, etc.) in RPLC-ESI-MS of proteins and peptides did not yield a single strong candidate that could generally replace TFA. The enhancement in sensitivity with other reagents observed in some cases strongly depended on the analyte, the experimental conditions used, and the mass spectrometer and, usually, it did not compensate for the loss in separation resolution related to TFA. The examples of direct coupling of affinity, size-exclusion, or ion-exchange chromatography (IEC) to ESI-MS are very limited because of incompatibilities related to the use of mobile phases containing high salt concentrations. To overcome this problem, an intermediate desalting step is needed. Multidimensional chromatography, microdialysis, and ion-capture modules can be used to couple these chromatographic modes with ESI-MS. Multidimensional chromatography with RPLC as a second dimension has most often been used. Although most examples involve the trap and analysis in the second dimension of a certain part of the first separation, some comprehensive analyses of the entire sample in the second dimension have also appeared. PMID- 16213446 TI - Protein glycosylation analysis by liquid chromatography-mass spectrometry. AB - Liquid chromatography (LC)-mass spectrometry (MS) has developed into an invaluable technology for the analysis of protein glycosylation. This review focuses on the recent developments in LC and combinations thereof with MS for this field of research. Recently introduced methods for the structural analysis of released glycans (native or derivatised) as well as glycopeptides, on normal phase, reverse phase and graphitized carbon LC columns with online MS(/MS) will be reviewed. Performed on nano-scale or capillary-scale, these LC-MS methods operate at femtomole sensitivity and support the further integration of glycosylation analysis in proteomics methodology. PMID- 16213447 TI - Narrow-bore sample trapping and chromatography combined with quadrupole/time-of flight mass spectrometry for ultra-sensitive identification of in vivo and in vitro metabolites. AB - The identification of in vitro and in vivo metabolites is vital to the discovery and development of new pharmaceutical therapies. Analytical strategies to identify metabolites at different stages of this process vary, but all involve the use of liquid chromatography separations combined with detection via mass spectrometry (HPLC/MS). Reported here is the use of narrow-bore column (0.5-1.0 mm i.d.) trapping of metabolites, followed by back-flushing onto a matching analytical column. Separated metabolites were then identified using quadrupole time-of-flight mass spectrometry (MS) and tandem MS. Metabolites in human plasma and from low-level in vitro incubations, that were not identified using standard HPLC/MS approaches, were characterized using the instrumental configuration described here. PMID- 16213448 TI - Mass spectrometric analysis of integral membrane proteins at the subpicomolar level: application to rhodopsin. AB - Integral membrane proteins are among the most interesting molecules for biomedical research, as some of the most important cellular functions are inherently tied to biological membranes. One such example is the vast expanse of receptors on cell surfaces. However, due to difficulties in the biochemical purification and structure/function analysis of membrane proteins, caused by their hydrophobic or amphophilic nature, membrane proteins are still much less studied than soluble proteins. Our laboratory has successfully developed and applied a methodology for the mass spectrometric analysis of integral membrane proteins. Here, we present an improvement in the sensitivity of detection made possible by the advancement of mass spectrometric instrumentation and refinement of the chromatographic analysis. Subpicomolar samples of bovine rhodopsin purified from native membranes were successfully analyzed, obtaining complete sequence coverage and the detection and localization of posttranslational modifications. Therefore, it is demonstrated that the detection limits and sequence coverage for soluble and membrane proteins can be comparable. The methodology presented here allows mass spectrometric analysis of subpicomolar levels of photopigments or other integral membrane proteins either from their native membranes or as products of expression systems. PMID- 16213450 TI - 2-hydrazino-1-methylpyridine: a highly sensitive derivatization reagent for oxosteroids in liquid chromatography-electrospray ionization-mass spectrometry. AB - A derivatization reagent, 2-hydrazino-1-methylpyridine, was developed for the liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) of oxosteroids. The reagent quantitatively reacted with oxosteroids at 60 degrees C within 1h and the resulting derivatives of the mono-oxosteroids provided a 70 1600-fold higher sensitivity compared to intact steroids. However, HMP was unsuitable for di-oxosteroids, such as androstenedione and progesterone. The developed derivatization procedure was applied to the LC-ESI-MS analysis of 5alpha-dihydrotestosterone in human prostate, and allowed the reproducible quantification of nanogram/gram level of the androgen with a 10-mg sample. PMID- 16213449 TI - Simple means to alleviate sensitivity loss by trifluoroacetic acid (TFA) mobile phases in the hydrophilic interaction chromatography-electrospray tandem mass spectrometric (HILIC-ESI/MS/MS) bioanalysis of basic compounds. AB - Trifluoroacetic acid (TFA) is a commonly used additive in HPLC and LC-MS analysis of basic compounds. It is also routinely added to aqueous-organic mobile phases utilized in the hydrophilic interaction chromatography-electrospray tandem mass spectrometry (HILIC-ESI/MS/MS) technique used in our laboratories for bioanalysis. However, TFA is known to suppress the ESI signals of analytes due to its ability to form gas-phase ion pairs with positively-charged analyte ions. The most common method to overcome this problem involves the post-column addition of a mixture of propionic acid and isopropanol. However the post-column addition setup requires additional pumps and is not desirable for continuous analysis of large amounts of samples. In this paper we present a simple yet very effective means of minimizing the negative effect of TFA in bioanalysis by direct addition of 0.5% acetic acid or 1% propionic acid to mobile phases containing either 0.025 or 0.05% TFA. A factor of two- to five-fold signal enhancement was achieved for eight basic compounds studied. Furthermore, chromatography integrity was maintained even with the addition of acetic acid and propionic acid to existing TFA mobile phases. This method has been successfully applied to the HILIC ESI/MS/MS high-throughput analysis of extracted biological samples to support pre clinical and clinical studies. PMID- 16213451 TI - Simultaneous determination of norethindrone and ethinyl estradiol in human plasma by high performance liquid chromatography with tandem mass spectrometry- experiences on developing a highly selective method using derivatization reagent for enhancing sensitivity. AB - In the present work, for the first time, a liquid chromatographic method with tandem mass spectrometric detection (LC-MS/MS) for the simultaneous analysis of norethindrone, and ethinyl estradiol, was developed and validated over the concentration range of 50-10000pg/ml and 2.5-500pg/ml, respectively, using 0.5 ml of plasma sample. Norethindrone, ethinyl estradiol, and their internal standards norethindrone-(13)C2, and ethinyl estradiol-d4, were extracted from human plasma matrix with n-butyl chloride. After evaporation of the organic solvent, the extract was derivatized with dansyl chloride and the mixture was injected onto the LC-MS/MS system. The gradient chromatographic elution was achieved on a Genesis RP-18 (50 mm x 4.6 mm, 3 microm) column with mobile phase consisted of acetonitrile, water and formic acid. The flow rate was 1.0 ml/min and the total run time was 5.0 min. Important parameters such as sensitivity, linearity, matrix effect, reproducibility, stability, carry-over and recovery were investigated during the validation. The inter-day precision and accuracy of the quality control samples at low, medium and high concentration levels were <6.8% relative standard deviation (RSD) and 4.4% relative error (RE) for norethindrone, and 4.2% RSD and 5.9% RE for ethinyl estradiol, respectively. Chromatographic conditions were optimized to separate analytes of interest from the potential interference peaks, arising from the derivatization. This method could be used for pharmacokinetic and drug-drug interaction studies in human subjects. PMID- 16213452 TI - Unusual properties of water at hydrophilic/hydrophobic interfaces. AB - The behaviour of water at mosaic hydrophilic/hydrophobic surfaces of different silicas and in biosystems (biomacromolecules, yeast cells, wheat seeds, bone and muscular tissues) was studied in different dispersion media over wide temperature range using 1H NMR spectroscopy with layer-by-layer freezing-out of bulk water (close to 273 K) and interfacial water (180 < T < 273 K), thermally stimulated depolarization current (TSDC) (90 < T < 270 K), infrared (IR) spectroscopy, and quantum chemical methods. Bulk water and water bound to hydrophilic/hydrophobic interfaces can be assigned to different structural types. There are (i) weakly associated interfacial water (1H NMR chemical shift delta(H) = 1.1-1.7 ppm) that can be assigned to high-density water (HDW) with collapsed structure (CS), representing individual molecules in hydrophobic pockets, small clusters and interstitial water with strongly distorted hydrogen bonds or without them, and (ii) strongly associated interfacial water (delta(H) = 4-5 ppm) with larger clusters, nano- and microdomains, and continuous interfacial layer with both HDW and low-density water (LDW). The molecular mobility of weakly associated bound water is higher (because hydrogen bonds are distorted and weakened and their number is smaller than that for strongly associated water) than that of strongly associated bound water (with strong hydrogen bonds but nevertheless weaker than that in ice Ih) that results in the difference in the temperature dependences of the 1H NMR spectra at T < 273 K. These different waters are also appear in changes in the IR and TSDC spectra. PMID- 16213453 TI - Surface-enhanced Raman scattering on colloidal nanostructures. AB - Surface-enhanced Raman scattering combines extremely high sensitivity, due to enhanced Raman cross-sections comparable or even better than fluorescence, with the observation of vibrational spectra of adsorbed species, providing one of the most incisive analytical methods for chemical and biochemical detection and analysis. SERS spectra are observed from a molecule-nanostructure enhancing system. This symbiosis molecule-nanostructure is a fertile ground for theoretical developments and a realm of applications from single molecule detection to biomedical diagnostic and techniques for nanostructure characterization. PMID- 16213454 TI - A quality-controlled microarray method for gene expression profiling. AB - Gene expression profiling on microarrays is widely used to measure the expression of large numbers of genes in a single experiment. Because of the high cost of this method, feasible numbers of replicates are limited, thus impairing the power of statistical analysis. As a step toward reducing technically induced variation, we developed a procedure of sample preparation and analysis that minimizes the number of sample manipulation steps, introduces quality control before array hybridization, and allows recovery of the prepared mRNA for independent validation of results. Sample preparation is based on mRNA separation using oligo(dT) magnetic beads, which are subsequently used for first-strand cDNA synthesis on the beads. cDNA covalently bound to the magnetic beads is used as template for second-strand cDNA synthesis, leaving the intact mRNA in solution for further analysis. The quality of the synthesized cDNA can be assessed by quantitative polymerase chain reaction using 3'- and 5'-specific primer pairs for housekeeping genes such as glyceraldehyde-3-phosphate dehydrogenase. Second strand cDNA is chemically labeled with fluorescent dyes to avoid dye bias in enzymatic labeling reactions. After hybridization of two differently labeled samples to microarray slides, arrays are scanned and images analyzed automatically with high reproducibility. Quantile-normalized data from five biological replica display a coefficient of variation 45% for 90% of profiled genes, allowing detection of twofold changes with false positive and false negative rates of 10% each. We demonstrate successful application of the procedure for expression profiling in plant leaf tissue. However, the method could be easily adapted for samples from animal including human or from microbial origin. PMID- 16213455 TI - Investigations of the effect of DNA size in transient transfection assay using dual luciferase system. AB - In transient transfection assays deletions, insertions, or truncations are broadly used to define cis regulatory elements, such as promoters, enhancers, and silencers. This application is based on the assumption that size changes of a test fragment have little or no effect on the apparent activity of the reporter genes. While it is known that unusually large size of a construct, such as PAC DNA, substantially reduces transfection efficiency, the size effect of a DNA fragment ranging from a couple of hundred basepairs to a few thousand basepairs, within which most transient transfection assays are performed, has not been rigorously investigated. Here we report a systematic study on the effect of plasmid size on the measurement of promoter/enhancer activity in transient transfection assay based on the dual luciferase reporter system. By inserting various lengths of stuffer DNA into the luciferase parental plasmids, we found an inverse correlation between the relative luciferase activity and the construct size. Furthermore, this size effect can be only partially corrected by inserting the same size of stuffer DNA into both experimental and control plasmids. The results suggest that the effect of DNA size should be taken into consideration for evaluation of transcriptional elements using transient transfection analysis. PMID- 16213456 TI - Determination of methotrexate, several pteridines, and creatinine in human urine, previous oxidation with potassium permanganate, using HPLC with photometric and fluorimetric serial detection. AB - A novel HPLC method, using UV and fluorimetric serial detection, for the simultaneous determination of methotrexate (MTX), five disease marker pteridines, and the reference metabolic subproduct creatinine (CREA) in human urine was established. A previous oxidation process using 10(-3) M KMnO4 (pH 5.0) and 35min of oxidation time was necessary to transform the analytes in the highly fluorescent pteridinic rings. CREA was not affected by the oxidative medium. Using Tris-HCl/NaCl buffer solution (pH 6.6) as mobile phase, MTX and the assayed pteridines were monitored by fluorescence at lambda(em) = 444 nm and lambda(ex) = 280 nm and creatinine was monitored by absorption measurements at lambda(abs) = 230 nm. All components were well resolved in approximately 7 min. Detection limits, according the criteria of Clayton and co-workers, were 10 ng ml(-1) for MTX, less than 1 ng ml(-1) for all of the pteridines, and 4 microg ml(-1) for CREA. PMID- 16213458 TI - Flow-injection enzymatic analysis for glycerol and triacylglycerol. AB - A flow-injection enzymatic analytical system was developed for determination of glycerol and triacylglycerol based on enzymatic reactions in capillary followed by electrochemical detection. The hydrogen peroxide produced from the enzyme reaction was monitored by a platinum-based electrochemical probe. Different immobilization strategies on silica support were studied. The best and most effective configuration found for the measurement of glycerol and triacylglycerols in this system was the tandem connection of a lipase column and a silica-fused capillary column coimmobilized with glycerokinase (GK) and glycerol-3-phosphate oxidase (GPO). Lipase helps the breakdown of triacylglycerol to yield free fatty acids and glycerol, while glycerokinase catalyzes the adenosine-5-triphosphate-dependent phosphorylation of glycerol to yield alpha glycerol phosphate, which can subsequently be oxidized by 3-glycerol phosphate oxidase to produce hydrogen peroxide. Response-surface methodology (RSM) was applied to optimize the proposed system for glycerol. Experiment settings were designed by central composite design to investigate the combined effects of pH, flow rate, reaction temperature, and ATP concentration on collected signals. The fitted model, per RSM, showed that the optimum conditions of the system are 2 mM ATP in 0.1 M carbonate buffer (pH 11.0), flow rate of 0.18 mL/min, temperature of 35 degrees C, 20 microL of sample injection, and applied voltage of 0.650 V. The proposed biosensing system using lipase, GK, and GPO exhibited a flow-injection analysis peak response of 2.5 min and a detection limit of 5 x 10(-5) M glycerol (S/N = 3) with acceptable reproducibility (CV < 4.30%). It also had linear working ranges from 10(-4) to 10(-2) M for glycerol and from 10(-3) to 10(-2) M for triacylglycerol. The capillary enzyme reactor was stable up to 2 months in continuous operation, and it was possible to analyze up to 15 samples per hour. The present biosensing system holds promise for on-line detection of triacylglycerol in serum and glycerol content in fermented products. PMID- 16213457 TI - Identification of actin as quercetin-binding protein: an approach to identify target molecules for specific ligands. AB - The biological effect of flavonoids is commonly studied by assaying the activity of a protein of interest. Taking a reverse approach, we identified target proteins of the widely studied flavonol quercetin by exploiting the altered spectroscopic properties of target proteins and ligands on their molecular interaction. Nuclear extracts of human leukemia cells were fractionated by column chromatography and assayed for their ability to alter the fluorescence emission spectra, and finally the proteins present in fractions of interest were identified by mass spectrometry. Among the identified proteins, actin was shown to be a quercetin-binding nuclear protein. PMID- 16213459 TI - Quantitative atomic force microscopy image analysis of unusual filaments formed by the Acanthamoeba castellanii myosin II rod domain. AB - We describe a quantitative analysis of Acanthamoeba castellanii myosin II rod domain images collected from atomic force microscope experiments. These images reveal that the rod domain forms a novel filament structure, most likely requiring unusual head-to-tail interactions. Similar filaments are seen also in negatively stained electron microscopy images. Truncated myosins from Acanthamoeba and other model organisms have been visualized before, revealing laterally associated bipolar minifilaments. In contrast, the filament structures that we observe are dominated by axial rather than lateral polymerization. The unusually small features in this structure (1-5 nm) required the development of quantitative and statistical techniques for filament image analysis. These techniques enhance the extraction of features that hitherto have been difficult to ascertain from more qualitative imaging approaches. The heights of the filaments are observed to have a bimodal distribution consistent with the diameters of a single rod domain and a pair of close-packed rod domains. Further quantitative analysis indicates that in-plane association is limited to at most a pair of rod domains. Taken together, this implies that the filaments contain no more than four rod domains laterally associated with one another, somewhat less than that seen in bipolar minifilaments. Analysis of images of the filaments decorated with an anti-FLAG antibody reveals head-to-tail association with mean distances between the antibodies of 75 +/- 15 nm. We consider a set of molecular models to help interpret possible structures of the filaments. PMID- 16213460 TI - Titrimetric method for determination of O-desulfated heparin in physiological samples using protamine-sensitive membrane electrode as endpoint detector. AB - O-Desulfated heparin (ODSH) is a promising new anti-inflammatory agent for the prevention of reperfusion injury following myocardial infarction or stroke. This partially desulfated heparin derivative has less anticoagulant activity than unfractionated heparin but retains the inherent anti-inflammatory properties of heparin. Thus, ODSH could be administered at the high doses needed to achieve desired anti-inflammatory function without risk of hemorrhage. However, given the very low anticoagulant activity of this species, traditional methods for heparin determination in clinical samples might not be well suited for ODSH measurements. In this article, a novel titrimetric method for detection of ODSH in buffer and plasma is described using a protamine-sensitive polymer membrane electrode as the detector. Titrations of ODSH with the heparin antagonist protamine yield sharp endpoints with sensitivity to ODSH in the micrograms per milliliter range for plasma samples. The stoichiometry for protamine interaction with ODSH is determined to average 1.39 microg protamine/microg ODSH in plasma. This technology is further applied to a toxicokinetic study of ODSH in an animal model, demonstrating the ability to detect the changes in ODSH concentrations in biological samples. PMID- 16213461 TI - Epigenetic regulation of mammalian pericentric heterochromatin in vivo by HP1. AB - We developed a model system whereby HP1 can be targeted to pericentric heterochromatin in ES cells lacking Suv(3)9h1/2 histone methyltransferase (HMTase) activities. HP1 so targeted can reconstitute tri-methylated lysine 9 of histone H3 (Me(3)K9H3) and tri-methylated lysine 20 of histone H4 (Me(3)K20H4) at pericentric heterochromatin, indicating that HP1 can regulate the distribution of these histone modifications in vivo. Both homo- and hetero-typic interactions between the HP1 isotypes were demonstrated in vivo as were HP1 interactions with the ESET/SETDB1 HMTase and the ATRX chromatin remodelling enzyme. We conclude that HP1 not only "deciphers" the histone code but can also "encode it". PMID- 16213462 TI - Inhibition of Atm and/or Atr disrupts gene silencing on the inactive X chromosome. AB - ATM and ATR are well documented for their roles in maintaining the integrity of genomic DNA by responding to DNA damage and preparing the cell for repair. Since ATM and ATR have been reported to exist in complexes with histone deacetylases, we asked whether Atm and Atr might also uphold gene silencing by heterochromatin. We show that the Atm/Atr inhibitor 2-aminopurine causes the inactive X chromosome to accumulate abnormal chromatin and undergo unwanted gene reactivation. We provide evidence that this gene expression from the inactive X chromosome is not a byproduct of the accumulation of DNA breaks. Individually inhibiting Atm and Atr by either small interfering RNA or the expression of dominant-negative ATM and ATR constructs also compromised X-inactivation. Atm and Atr, therefore, not only function in responding to DNA damage but perhaps also are involved in gene silencing via the maintenance of heterochromatin. PMID- 16213463 TI - Overexpression of TLR2 and TLR4 susceptibility to serum deprivation-induced apoptosis in CHO cells. AB - We examined the effect of overexpression of TLR2 and TLR4 on apoptosis. TLR2 and TLR4 transfected CHO cells were subjected to serum deprivation for 0, 24, and 48 h. CHO cells served as control. The survival was 80.4% and 66.8% in CHO cells, 73.8% and 47.6% in TLR2/CHO, and 70.5% and 53.0% in TLR4/CHO, respectively. Flow cytometry examination suggested that apoptotic cells were 7.17% and 32.91% in control CHO cells, 29.0% and 64.6% in TLR2/CHO, and 41.4% and 64.6% in TLR4/CHO, respectively. The levels of FasL and caspase-8 activity in TLR2/CHO and TLR4/CHO cells were significantly higher than that of CHO cells. Transfection of dominant negative FADD into TLR2/CHO and TLR4/CHO cells significantly reduced apoptosis. Our results suggest that overexpression of TLR2 and TLR4 in CHO cells sensitizes the cells to serum deprivation-induced apoptosis and that the mechanisms are involved in the death receptor-mediated signaling pathway. PMID- 16213464 TI - Novel time-dependent vascular actions of Delta9-tetrahydrocannabinol mediated by peroxisome proliferator-activated receptor gamma. AB - Cannabinoids have widespread effects on the cardiovascular system, only some of which are mediated via G-protein-coupled cell surface receptors. The active ingredient of cannabis, Delta9-tetrahydrocannabinol (THC), causes acute vasorelaxation in various arteries. Here we show for the first time that THC also causes slowly developing vasorelaxation through activation of peroxisome proliferator-activated receptors gamma (PPARgamma). In vitro, THC (10 microM) caused time-dependent vasorelaxation of rat isolated arteries. Time-dependent vasorelaxation to THC was similar to that produced by the PPARgamma agonist rosiglitazone and was inhibited by the PPARgamma antagonist GW9662 (1 microM), but not the cannabinoid CB1 receptor antagonist AM251 (1 microM). Time-dependent vasorelaxation to THC requires an intact endothelium, nitric oxide, production of hydrogen peroxide, and de novo protein synthesis. In transactivation assays in cultured HEK293 cells, THC-activated PPARgamma, transiently expressed in combination with retinoid X receptor alpha and a luciferase reporter gene, in a concentration-dependent manner (100 nM-10 microM). In vitro incubation with THC (1 or 10 microM, 8 days) stimulated adipocyte differentiation in cultured 3T3L1 cells, a well-accepted property of PPARgamma ligands. The present results provide strong evidence that THC is a PPARgamma ligand, stimulation of which causes time dependent vasorelaxation, implying some of the pleiotropic effects of cannabis may be mediated by nuclear receptors. PMID- 16213465 TI - Momordin I, an inhibitor of AP-1, suppressed osteoclastogenesis through inhibition of NF-kappaB and AP-1 and also reduced osteoclast activity and survival. AB - Osteoclasts originating from hematopoietic precursor cells differentiate into multinucleated cells through multiple steps. The essential roles of NF-kappaB and AP-1 in osteoclast differentiation have been clearly demonstrated in numerous studies. c-Fos, a component of AP-1 transcription factor, plays a key role in osteoclast differentiation. Recently, we found a strong inhibitor of AP-1 transcriptional activity, named momordin I, based on the structure of oleanolic acid glycosides and originally isolated from Ampelopsis radix. So, we hypothesized that momordin I might be able to regulate osteoclast formation, activity, and survival. Here, we report the ability of momordin I to suppress osteoclastogenesis in a co-cultured system and a RANKL-induced osteoclast precursor system. Momordin I remarkably inhibited the activation of NF-kappaB as well as AP-1 in RANKL-induced RAW264.7 cells, in which momordin I appeared to target IkappaB degradation and c-Fos expression, respectively, but not MAPK signaling pathways. The ability of momordin I to change the ratio of RANKL and OPG in primary osteoblasts was partially responsible for the reduction of osteoclast formation. Furthermore, pit formation on dentin slices was suppressed by momordin I with stimulating actin ring disruption. Our results also showed that momordin I highly shortened osteoclast lifespan and induced osteoclast apoptosis. In conclusion, the present results demonstrate for the first time that momordin I is a potent inhibitor of osteoclast differentiation via the reduction of NF-kappaB and AP-1, and also suppresses osteoclast function and survival. PMID- 16213466 TI - Predicting protein subnuclear location with optimized evidence-theoretic K nearest classifier and pseudo amino acid composition. AB - The nucleus is the brain of eukaryotic cells that guides the life processes of the cell by issuing key instructions. For in-depth understanding of the biochemical process of the nucleus, the knowledge of localization of nuclear proteins is very important. With the avalanche of protein sequences generated in the post-genomic era, it is highly desired to develop an automated method for fast annotating the subnuclear locations for numerous newly found nuclear protein sequences so as to be able to timely utilize them for basic research and drug discovery. In view of this, a novel approach is developed for predicting the protein subnuclear location. It is featured by introducing a powerful classifier, the optimized evidence-theoretic K-nearest classifier, and using the pseudo amino acid composition [K.C. Chou, PROTEINS: Structure, Function, and Genetics, 43 (2001) 246], which can incorporate a considerable amount of sequence-order effects, to represent protein samples. As a demonstration, identifications were performed for 370 nuclear proteins among the following 9 subnuclear locations: (1) Cajal body, (2) chromatin, (3) heterochromatin, (4) nuclear diffuse, (5) nuclear pore, (6) nuclear speckle, (7) nucleolus, (8) PcG body, and (9) PML body. The overall success rates thus obtained by both the re-substitution test and jackknife cross-validation test are significantly higher than those by existing classifiers on the same working dataset. It is anticipated that the powerful approach may also become a useful high throughput vehicle to bridge the huge gap occurring in the post-genomic era between the number of gene sequences in databases and the number of gene products that have been functionally characterized. The OET-KNN classifier will be available at www.pami.sjtu.edu.cn/people/hbshen. PMID- 16213467 TI - Butyrylcholinesterase, paraoxonase, and albumin esterase, but not carboxylesterase, are present in human plasma. AB - The goal of this work was to identify the esterases in human plasma and to clarify common misconceptions. The method for identifying esterases was nondenaturing gradient gel electrophoresis stained for esterase activity. We report that human plasma contains four esterases: butyrylcholinesterase (EC 3.1.1.8), paraoxonase (EC 3.1.8.1), acetylcholinesterase (EC 3.1.1.7), and albumin. Butyrylcholinesterase (BChE), paraoxonase (PON1), and albumin are in high enough concentrations to contribute significantly to ester hydrolysis. However, only trace amounts of acetylcholinesterase (AChE) are present. Monomeric AChE is seen in wild-type as well as in silent BChE plasma. Albumin has esterase activity with alpha- and beta-naphthylacetate as well as with p-nitrophenyl acetate. Misconception #1 is that human plasma contains carboxylesterase. We demonstrate that human plasma contains no carboxylesterase (EC 3.1.1.1), in contrast to mouse, rat, rabbit, horse, cat, and tiger that have high amounts of plasma carboxylesterase. Misconception #2 is that lab animals have BChE but no AChE in their plasma. We demonstrate that mice, unlike humans, have substantial amounts of soluble AChE as well as BChE in their plasma. Plasma from AChE and BChE knockout mice allowed identification of AChE and BChE bands without the use of inhibitors. Human BChE is irreversibly inhibited by diisopropylfluorophosphate, echothiophate, and paraoxon, but mouse BChE spontaneously reactivates. Since human plasma contains no carboxylesterase, only BChE, PON1, and albumin esterases need to be considered when evaluating hydrolysis of an ester drug in human plasma. PMID- 16213468 TI - Stress differentially modulates fear conditioning in healthy men and women. AB - BACKGROUND: Stress and stress hormones modulate emotional learning in rats and might have similar effects in humans. Theoretic accounts of posttraumatic stress disorder (PTSD), for example, implicate the stress-induced modulation of fear conditioning in the development of intrusive emotional reactions. The present study examined the impact of acute stress and cortisol (CORT) on classically conditioned fear in men and women. METHODS: Ninety-four healthy undergraduates were exposed to a mild stressor (or control condition) while subjective anxiety and glucocorticoid stress responses (salivary CORT) were measured. One hour later, all participants participated in a differential fear conditioning procedure while conditioned skin conductance responses (SCR) were recorded. RESULTS: Exposure to the stressor increased subjective anxiety and elevated CORT levels. In men, stress exposure facilitated fear conditioning; whereas in women, stress appeared to inhibit fear conditioning. The impact of stress on differential conditioning in men was associated with increased CORT levels. CONCLUSIONS: Consistent with animal models, these results demonstrate that stress exposure can modulate classical conditioning in humans, possibly via hormonal mechanisms. The enhancing effects of stress on the formation of conditioned fear might provide a useful model for the formation of pathological emotional reactions, such as those found in PTSD. PMID- 16213469 TI - A polymorphism in the PDLIM5 gene associated with gene expression and schizophrenia. AB - BACKGROUND: Results of recent DNA microarray analyses of postmortem brains of patients with schizophrenia revealed that expression of the PDZ and LIM domain 5 gene (PDLIM5) is increased. In the present study, we examined whether polymorphisms in PDLIM5 are associated with schizophrenia. METHODS: We screened for mutations in PDLIM5 in 24 Japanese patients with schizophrenia and evaluated the associations of the identified polymorphisms with schizophrenia in a Japanese case-control population (total samples, 278 schizophrenia patients and 462 control subjects). Expression of PDLIM5 was quantified by real-time quantitative polymerase chain reaction (PCR) in postmortem prefrontal cortex of 34 schizophrenia patients. Electrophoretic mobility shift assay (EMSA) was performed to examine whether a polymorphism influences nuclear protein binding. RESULTS: We identified 27 polymorphisms in PDLIM5 and found associations between polymorphisms (rs2433320 and rs2433322) in the 5' region of the gene and schizophrenia (p = .004). Real-time quantitative PCR revealed that these polymorphisms influenced gene expression (p = .007). An EMSA showed that the different alleles of the rs2433320 polymorphism bound differently to nuclear proteins. CONCLUSIONS: These results suggest that PDLIM5 might play a role in genetic susceptibility to schizophrenia. PMID- 16213470 TI - Lack of effects on core obsessive-compulsive symptoms of tryptophan depletion during symptom provocation in remitted obsessive-compulsive disorder patients. AB - BACKGROUND: Pharmacological evidence support that enhancement of serotonin (5-HT) neurotransmission is critical for treatment efficacy in obsessive-compulsive disorder (OCD). Surprisingly, acute tryptophan depletion (ATD), a procedure known to reduce 5-HT neurotransmission, carried out in remitted OCD patients on selective serotonin reuptake inhibitors (SSRIs) failed to worsen obsessive compulsive (OC) symptoms. We hypothesized that the putative symptom exacerbation resulting from ATD would only be observed during symptom provocation but not at rest. METHODS: Double-blind placebo-controlled ATD study conducted in 16 OCD patients with stable improvement under either SSRI (n = 8) or specialized cognitive behavior therapy alone (n = 8), coupled with gradual symptom provocation, performed 5 hours after drink ingestion. RESULTS: Acute tryptophan depletion markedly reduced total and free plasma tryptophan levels but did not significantly increase obsessions or compulsions at rest or following symptom provocation. However, subjective distress in response to triggering situations was significantly higher during ATD; significant mood lowering was also present during ATD. CONCLUSIONS: These results are consistent with the view that relapses in OC core symptoms in remitted OCD patients may not depend solely on short-term changes in presynaptic 5-HT availability. In contrast to its apparent lack of effect on core OC symptoms, ATD affected the patient's mood and distress level resulting from provocation. PMID- 16213471 TI - Prefrontal cortex and amygdala volume in first minor or major depressive episode after cancer diagnosis. AB - BACKGROUND: Major and minor depressive episodes in cancer patients are frequent and are frequently seen as the first depressive episode in a patient's life. However, the neurological basis of these depressive episodes remains largely unknown. METHODS: Subjects were 51 breast cancer survivors (BCS) who had no history of any depressive episode before the cancer diagnosis (11 BCS with a history of a first minor depressive episode after cancer diagnosis, 11 BCS with a history of a first major depressive episode after cancer diagnosis, and 29 BCS with no history of any depressive episode after cancer diagnosis). We analyzed the prefrontal cortex (PFC) and amygdala volumes in a 1.5-Tesla Magnetic Resonance Imaging scanner. We characterized the structural correlates of depression using two complementary approaches. The first was voxel-based morphometry (VBM) that allowed us to scan the entire brain for reactive gray matter deficit. The second was classical volumetry focusing on the amygdala. RESULTS: Voxel-based morphometry revealed no brain region, including PFC, for which volume was significantly different among the three groups. There were trend level differences in the left amygdala volume in the manual tracing method among the three groups. The left amygdala volumes in the subjects with a first minor and/or major depressive episode were significantly smaller than in those with no history of any depressive episode. CONCLUSIONS: It might be suggested that amygdala volume was associated with a first minor and/or major depressive episode after cancer diagnosis. PMID- 16213472 TI - Event-related oscillations in offspring of alcoholics: neurocognitive disinhibition as a risk for alcoholism. AB - BACKGROUND: Event-related oscillations (EROs) are increasingly being used to assess neurocognitive functioning in normal and clinical populations. The current study compares different frequency activities in offspring of alcoholics (OA) and in normal control subjects (NC) to examine whether the OA group exhibits any abnormality in oscillatory activity while performing a Go/NoGo task. METHODS: The S-transform algorithm was employed to decompose the electroencephalographic (EEG) signals into different time-frequency bands, and the oscillatory responses in the P300 time window (300-700 milliseconds) were statistically analyzed in both groups. RESULTS: The OA group manifested significantly decreased activity in delta (1-3 Hz), theta (4-7 Hz), and alpha1 (8-9 Hz) bands during the NoGo condition, as well as reduced delta and theta activity during the Go condition. This reduction was more prominent in the NoGo than in the Go condition. CONCLUSIONS: The decreased response in delta, theta, and alpha1 oscillations, especially during the NoGo condition in high-risk individuals, is perhaps suggestive of cognitive and neural disinhibition and may serve as an endophenotypic marker in the development of alcoholism and/or other disinhibitory disorders. PMID- 16213473 TI - Acute occupancy of brain serotonin transporter by sertraline as measured by [11C]DASB and positron emission tomography. AB - BACKGROUND: In vivo determination of serotonin transporter (5-HTT) occupancy by selective serotonin reuptake inhibitors (SSRI) using positron emission tomography (PET) can aid in determination of dosing. Previous studies used chronic SSRI administration that may down-regulate 5-HTT and used the cerebellum as reference region despite measurable 5-HTT. We examine the reference region and occupancy after acute sertraline dosing. METHODS: We conducted autoradiography of human postmortem cerebellum to determine an optimal reference region. We quantified 5 HTT binding using [(11)C]DASB and arterial input functions in 17 healthy volunteers. Baseline PET scans were followed by a scan 4-6 days after 25 mg to 100mg of daily sertraline. Several modeling methods and outcome measures were assessed. RESULTS: Cerebellar gray matter is the optimal reference region. Occupation of 5-HTT sites saturates at low plasma sertraline levels (K(D) = 1.9 ng/ml) with maximal occupancies of 106.8 +/- 8.3% across all brain regions. There is a weak correlation between oral sertraline and plasma sertraline. Occupancy measures vary based on the reference region and outcome measure used. CONCLUSIONS: Occupancy studies and postmortem autoradiography can help define the optimal reference region. Reference tissue modeling using the optimal reference region returns the same occupancy measures as those determined using an arterial input function. PMID- 16213474 TI - Matrix metalloproteinase-2 and -9 are induced differently by doxorubicin in H9c2 cells: The role of MAP kinases and NAD(P)H oxidase. AB - OBJECTIVE: Dysregulation of myocardial metalloproteinases (MMPs) is now regarded as an early contributory mechanism for the initiation and progression of heart failure. Doxorubicin is a strongly cardiotoxic anticancer drug. This study investigates the effects of doxorubicin on myocardial MMP-2 and MMP-9 activation. METHODS: After pre-treatment with or without carvedilol or dexrazoxane, we exposed H9c2 cardiomyocytes to doxorubicin to evaluate reactive oxygen species (ROS) formation and MMP-2 and MMP-9 expression and activation. To investigate the signaling pathways leading to doxorubicin-induced MMP activation, we also examined the phosphorylation of three members of the MAPK family (ERK1/2, p38, and JNK), the effects of selective inhibitors of ERK1/2, p38, and JNK on MMP transcription and activity, the transcription of the NAD(P)H oxidase subunit Nox1, and the effects of the NAD(P)H oxidase inhibitor DPI on MMP activation. RESULTS: Doxorubicin induces a significant increase in ROS formation and a rapid increase of MMP expression and activation. Pre-treatment with carvedilol or dexrazoxane prevented these effects. We also found that p38 is the MAPK that is mainly responsible for MMP-9 activation through an NAD(P)H-independent mechanism. ERK and JNK modulate the transcription of the NAD(P)H oxidase subunit Nox1, while the JNK/ERK NAD(P)H oxidase cascade is an important pathway that mediates doxorubicin signaling to MMP-2. Inhibition of NAD(P)H oxidase attenuates the increase in MMP-2, but augments the doxorubicin-induced increase in MMP-9. CONCLUSIONS: Enhancement of MMP-2 and MMP-9 in cardiac myocytes in response to doxorubicin is mediated by the cooperation of ERK, JNK, and p38 kinase pathways, most of which are redox dependent. PMID- 16213475 TI - Autophagy in cardiac myocyte homeostasis, aging, and pathology. AB - Autophagy, an intralysosomal degradation of cells' own constituents that includes macro-, micro-, and chaperone-mediated autophagy, plays an important role in the renewal of cardiac myocytes. This cell type is represented by long-lived postmitotic cells with very poor (if any) replacement through differentiation of stem cells. Macroautophagy, the most universal form of autophagy, is responsible for the degradation of various macromolecules and organelles including mitochondria and is activated in response to stress, promoting cell survival. This process is also involved in programmed cell death when injury is irreversible. Even under normal conditions, autophagy is somewhat imperfect, underlying gradual accumulation of defective mitochondria and lipofuscin granules within aging cardiac myocytes. Autophagy is involved in the most important cardiac pathologies including myocardial hypertrophy, cardiomyopathies, and ischemic heart disease, a fact that has led to increasing attention to this process. PMID- 16213476 TI - Immune cell activation and subsequent epithelial dysfunction by Staphylococcus enterotoxin B is attenuated by the green tea polyphenol (-)-epigallocatechin gallate. AB - Bacterial superantigens (SAg) are potent T cell activators and when delivered systemically elicit a self-limiting enteropathy in mice. Also, SAg-stimulated human peripheral blood mononuclear cells (PBMC) increase enteric epithelial cell monolayer permeability in vitro. Epigallocatechin gallate (EGCG), the major polyphenol component of green tea (Camilla sinesis) leaf, has been presented as an anti-inflammatory agent. We tested the hypothesis that EGCG (10-100 microM) would block PBMC activation by the SAg, Staphylococcus aureus enterotoxin B (SEB, 1 microg/ml), thus preventing disruption of the epithelial barrier. Pretreatment or co-treatment of human PBMC or murine lymphnode cells with EGCG significantly reduced SEB-induced proliferation and IL-2, IFNgamma, and TNFalpha production. ConA-induced proliferation was also inhibited by EGCG (50 microM) co-treatment. These effects of EGCG were not due to induction of immune cell apoptosis, and were independent of EGCGs anti-oxidant activity, and inhibition of NF-kappaB or AP-1 activation. Moreover, addition of exogenous IL-2 (20 ng/ml) to the cultures could not overcome the immunosuppressive effect of EGCG. Culture supernatant from PBMC stimulated in the presence of EGCG failed to increase the permeability of T84 epithelial cell monolayers: a finding consistent with the reduced IFNgamma and TNFalpha production by SAg+EGCG treated PBMC. These data promote EGCG as a suppressor of T cell activation, and given the prominent role that bacteria and T cells play in inflammatory disease we suggest that EGCG could be a useful addition to current treatments for enteric immune disorders and T cell driven immunopathologies. PMID- 16213477 TI - Constitutive expression of functional CD40 on mouse renal cancer cells: induction of Fas and Fas-mediated killing by CD40L. AB - CD40, a member of the TNF receptor superfamily, is expressed on B cells, dendritic cells, and some tumor cells, including melanoma and bladder carcinoma. In this study, we report that both mouse and human renal carcinoma cells (RCC) also constitutively express functional CD40. Treatment of mouse RCC with CD40L induced strong expression of genes and proteins for ICAM-1 and Fas, and this expression was further enhanced by combining CD40L with IFN-gamma. Similar effects were demonstrated using an agonist anti-CD40 antibody. The increased levels of Fas expression on RCC after treatment with CD40L plus IFN-gamma resulted in potent killing by either FasL-positive effector cells or agonistic anti-Fas antibody. The combination of CD40L plus IFN-gamma also significantly enhanced killing of RCC by tumor-specific CTL lines. Our results demonstrate that constitutively expressed CD40 is functionally active and may provide a molecular target for the development of new approaches to the treatment of RCC. PMID- 16213478 TI - High cardiovascular risk in young Saudi males: cardiovascular risk factors, diet and inflammatory markers. AB - BACKGROUND: The relationship between coronary risk score (CRS), individual coronary risk factors and the serum inflammatory markers, high sensitivity C reactive protein (hsCRP), ceruloplasmin (Cp), and soluble intercellular adhesion molecule-1 (sICAM-1) was studied in 140 Saudi males without clinically evident coronary heart disease (CHD). METHODS: One hundred forty subjects without clinically evident CHD were categorized into age tertiles. Demographic data together with an estimate of CRS using Framingham and PROCAM algorithms were obtained, and serum lipid profile, glucose, hsCRP, sICAM-1, and Cp were measured. Macronutrient intake was assessed by a questionnaire. The relationship between CRS, biochemical markers and diet was assessed by univariate and multivariate analysis. RESULTS: There was no significant difference in median hsCRP, sICAM-1 or Cp between the age groups. Serum Cp was positively associated with age (r=0.224, p<0.01) and FRS score (r=0.174, p<0.05). Serum sICAM-1 was negatively associated with PROCAM score (r=-0.183, p<0.05). sICAM-1 was positively associated with HDL cholesterol (r=0.36, p<0.0001) among non-diabetics and negatively associated (r=-0.397, p<0.05) among diabetic subjects. Age and dietary intake of saturated fatty acids together explained 7.9% of the variation in serum Cp level in a stepwise multiple regression model. Similarly 6.5% of the variation in serum sICAM-1 level was explained by the total cholesterol/HDL-C ratio. The youngest tertile of the group (<30 y) had the highest dietary intake of energy, fat and saturated fatty acids (p<0.05), and also had a high prevalence of obesity, smoking and sedentary lifestyle. CONCLUSION: We have demonstrated that there is a high prevalence of coronary risk factors and poor dietary intake within a Saudi male population, and that dietary factors are associated with serum sICAM-1 and ceruloplasmin but not hsCRP concentrations in this group. PMID- 16213479 TI - Reductions in blood lead levels among school children following the introduction of unleaded petrol in South Africa. AB - Epidemiological studies have indicated that in the 1980s and early 1990s (a period in which petrol lead concentrations in South Africa ranged from 0.836 to 0.4 g/L), large proportions of urban South African children were at risk of excessive exposure to environmental lead. In 1991, when the maximum permissible petrol lead concentration in the country equaled 0.4 g/L, a study determined that the mean blood lead level among children attending inner city schools in the Cape Peninsula equaled 16 microg/dL, with well over 90% of children having blood lead levels equaling or exceeding the internationally accepted guideline level of 10 microg/dL. Socio economic status, housing conditions, and proximity of children's schools and homes to heavily trafficked roads were among the factors significantly associated with blood lead concentrations. In 1996, unleaded petrol was introduced in South Africa. A study undertaken in 2002 (at the same schools as in 1991), when unleaded petrol constituted around 30% of the market share of petrol in the country, has shown significant reductions in the mean blood lead concentration among Cape Peninsula inner city children and in the proportion of children with elevated blood lead levels. The mean blood lead level for the total sample (n = 429) of children whose mean age equaled 7 years (range: 5-11 years) was 6.4 microg/dL (range: 1.0-24.5 microg/dL) and 10% of children had blood lead levels equalling or exceeding 10 microg/dL. The mean blood lead levels among children attending schools in an inner city and in a less heavily trafficked periurban suburb were 6.9 and 4.8 microg/dL, respectively. PMID- 16213480 TI - (+)-Cyclazosin, a selective alpha1B-adrenoceptor antagonist: functional evaluation in rat and rabbit tissues. AB - To shed light on the discrepancy between reported binding and functional affinity and selectivity at alpha(1b/B)-adrenoceptors, the antagonist (+)-cyclazosin was reinvestigated in rat and rabbit tissues. It displayed a competitive antagonism at alpha(1A) and alpha(1D)-adrenoceptors of rat prostatic vas deferens and aorta with pA(2) values 7.75 and 7.27, respectively. In rabbit thoracic aorta (+) cyclazosin competitively antagonized noradrenaline-induced contractions at alpha(1B)-adrenoceptors with a pA(2) value of 8.85, whereas its affinity at alpha(1L)-adrenoceptors was markedly lower (pA(2) = 6.75-7.09). In conclusion, these data confirmed that (+)-cyclazosin is a selective alpha(1B)-adrenoceptor antagonist also in functional assays, showing 13- and 38-fold selectivity for the alpha(1B)-adrenoceptor over alpha(1A)- and alpha(1D)-subtypes, respectively. Furthermore, (+)-cyclazosin displayed a significant selectivity for alpha(1B) adrenoceptors relative to the alpha(1L)-subtype. PMID- 16213483 TI - Accelerated recovery of 5-fluorouracil-damaged bone marrow after rosiglitazone treatment. AB - Our preliminary data indicate that rosiglitazone may be myeloprotective. We investigated whether it can modify bone marrow recovery. Five-day pre-treatment with rosiglitazone significantly accelerated recovery of 5-fluorouracil-damaged bone marrow in mice. Frequency and femoral content of granulocyte-macrophage progenitors reached mean baseline faster in pre-treated groups than in 5 fluorouracil-treated controls. Consequently, neutropenia was milder. Five-day insulin pre-treatment had similar effects in vivo. Insulin supports in vitro hematopoiesis. The observed myeloprotection demonstrated the importance of insulin in vivo. Clinical use of insulin to moderate myelotoxicity is impractical but rosiglitazone, an insulin sensitizer, could offer hope. Although rosiglitazone tends to increase plasma insulin levels, the significant myeloprotection was partly due to direct effects on progenitors. In vitro rosiglitazone enhanced the survival of both murine progenitor and human mobilized blood stem cells in the presence of 5-fluorouracil, the effect of which was neutralized by a peroxisome-proliferator-activated receptor-gamma antagonist. PMID- 16213481 TI - Inhibitory effects of histamine H4 receptor antagonists on experimental colitis in the rat. AB - The histamine H(4) receptor is a G-protein coupled receptor with little homology to the pro-inflammatory histamine H(1) receptor, expressed on cells of the immune system with hematopoietic lineage such as eosinophils and mast cells. The effects of the recently described highly selective histamine H(4) receptor antagonists JNJ 10191584 and JNJ 7777120 have now been investigated on the acute colitis provoked by trinitrobenzene sulphonic acid over 3 days in the rat. Treatment with JNJ 10191584 (10-100 mg/kg p.o., b.i.d.) caused a dose-dependent reduction in macroscopic damage, inhibition of the TNBS-provoked elevation of both colonic myeloperoxidase and tumour necrosis factor-alpha (TNF-alpha), and a reduction in the histologically assessed increase in mucosal and submucosal thickness and neutrophil infiltration. JNJ 7777120 (100 mg/kg p.o., b.i.d.) likewise reduced the macroscopic injury and the increases in colonic myeloperoxidase and TNF-alpha levels. These findings indicate a pro-inflammatory role for the histamine H(4) receptor in this model and suggest a novel pharmacological approach to the treatment of colitis. PMID- 16213482 TI - Central administration of small interfering RNAs in rats: a comparison with antisense oligonucleotides. AB - To date there are only few reports of the use of small interfering RNA (siRNA) in whole animals and most of these are restricted to systemic application of siRNAs targeting the liver. In our present studies we have investigated whether siRNAs can be used against a central target after intracerebroventricular (i.c.v.) application and compared their effects with those of antisense oligonucleotides. For this purpose we designed different siRNA and antisense oligonucleotide molecules against the rat hypothalamic melanocortin MC(4) receptor and selected the siRNA and antisense oligonucleotide with the highest efficacy in vitro. We observed that siRNA, encompassing the same gene sequence as antisense oligonucleotides, induced a stronger inhibition of melanocortin MC(4) receptor expression than antisense oligonucleotides. When fluorescence-labeled siRNA were applied i.c.v. in rats no label was detected in brain tissue in spite of the use of cell detergents to improve the delivery. In contrast to these findings the i.c.v. administered fluorescence-labeled antisense oligonucleotides reached the brain structures expressing melanocortin MC(4) receptor and were taken up by the cells in these areas. In summary it seems as if 'naked' antisense oligonucleotides have an advantage over 'naked' siRNA for experiments in vivo. The development of optimized vector systems seems to be a prerequisite before siRNA can be regarded as a suitable experimental tool for in vivo studies. PMID- 16213484 TI - Morphological integration and functional assessment of transplanted neural progenitor cells in healthy and acute ischemic rat eyes. AB - We have functionally and morphologically characterized the retina and optic nerve after neural progenitor cell transplants to healthy rat eyes and eyes damaged by acute elevation of intraocular pressure (IOP). Green fluorescent protein expressing adult rat hippocampal progenitor cells (AHPCs) were transplanted by intravitreal injection into healthy eyes and eyes damaged with acute ocular hypertension. Pupil light reflexes (PLR) and electroretinograms (ERGs) were recorded preoperatively and postoperatively. Eyes were subsequently prepared for immunohistochemical analysis and confocal imaging. Transplanted AHPCs were found in 8 of 15 (53%) acute ischemic eyes 62 days after surgery and 5 of 10 (50%) healthy eyes 32 days after grafting. Analysis of PLR and ERG function in acute ischemic eyes revealed no statistically significant difference compared to controls after transplantation for all observed functional parameters. Transplant into healthy rat eyes revealed no PLR or ERG amplitude deficits between transplanted and non-transplanted (control) eyes. Morphological and immunohistochemical analysis revealed that transplanted AHPCs survived and differentiated in both normal and injured retinal environments. Morphological integration occurred primarily within the inner retinal layers of the acute ischemic eyes. AHPCs were found to express neuronal and glial markers following transplantation. Transplanted AHPCs have the ability to integrate and differentiate in ischemia damaged retinas. PLR and ERG analysis revealed no significant difference in functional outcome in transplant recipient eyes. PMID- 16213485 TI - Focus on molecules: lumican. PMID- 16213486 TI - Selective regulation of cellular processes via protein cascades acting as band pass filters for time-limited oscillations. AB - We show by mathematical modelling that a two-level protein cascade can act as a band-pass filter for time-limited oscillations. The band-pass filters are then combined into a network of three-level signalling cascades that by filtering the frequency of time-limited oscillations selectively switches cellular processes on and off. The physiological relevance for the selective regulation of cellular processes is demonstrated for the case of regulation by time-limited calcium oscillations. PMID- 16213488 TI - Identification of a novel bean alpha-amylase inhibitor with chitinolytic activity. AB - Zabrotes subfasciatus is a devastating starch-dependent storage bean pest. In this study, we attempted to identify novel alpha-amylase inhibitors from wild bean seeds, with efficiency toward pest alpha-amylases. An inhibitor named Phaseolus vulgaris chitinolytic alpha-amylase inhibitor (PvCAI) was purified and mass spectrometry analyses showed a protein with 33330 Da with the ability to form dimers. Purified PvCAI showed significant inhibitory activity against larval Z. subfasciatus alpha-amylases with no activity against mammalian enzymes. N terminal sequence analyses showed an unexpected high identity to plant chitinases from the glycoside hydrolase family 18. Furthermore, their chitinolytic activity was also detected. Our data provides compelling evidence that PvCAI also possessed chitinolytic activity, indicating the emergence of a novel alpha amylase inhibitor class. PMID- 16213487 TI - The origin of the high sensitivity of muscle fructose 1,6-bisphosphatase towards AMP. AB - Adenosine 5'-monophosphate (AMP) inhibits muscle fructose 1,6-bisphosphatase (FBPase) about 44 times stronger than the liver isozyme. The key role in strong AMP binding to muscle isozyme play K20, T177 and Q179. Muscle FBPase which has been mutated towards the liver enzyme (K20E/T177M/Q179C) is inhibited by AMP about 26 times weaker than the wild-type muscle enzyme, but it binds the fluorescent AMP analogue, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5' monophosphate (TNP-AMP), similarly to the wild-type liver enzyme. The reverse mutation of liver FBPase towards the muscle isozyme significantly increases the affinity of the mutant to TNP-AMP. High affinity to the inhibitor but low sensitivity to AMP of the liver triple mutant suggest differences between the isozymes in the mechanism of allosteric signal transmission. PMID- 16213489 TI - ADAM12-mediated focal adhesion formation is differently regulated by beta1 and beta3 integrins. AB - ADAM12, adisintegrin and metalloprotease, has been demonstrated to be upregulated in human malignant tumors and to accelerate the malignant phenotype in a mouse model for breast cancer. ADAM12 is a substrate for beta1 integrins and may affect tumor and stromal cell behavior through its binding to beta1 integrins. Here, we report that cells deficient in beta1 integrin or overexpressing beta3 integrin can bind to recombinant full-length human ADAM12 via beta3 integrin. Furthermore, cell binding to ADAM12 via beta3 integrin results in the formation of focal adhesions, which are not formed upon beta1 integrin-mediated cell attachment. We also show that RhoA is involved in beta3 integrin-mediated focal adhesion formation. PMID- 16213491 TI - X-linked adrenoleukodystrophy mice demonstrate abnormalities in cholesterol metabolism. AB - The neurodegenerative disorder X-linked adrenoleukodystrophy (X-ALD) is caused by ABCD1 mutations and characterized by very long-chain fatty acid (VLCFA) accumulation. Cholesterol-lowering normalized VLCFA in fibroblasts and plasma of X-ALD patients. We show that in cultured cells, cholesterol-loading induces ABCD1. In X-ALD mice, plasma cholesterol is elevated and not further increasable by cholesterol-feeding, whereas hepatic HMG-CoA reductase and Abcd2 are downregulated. Upon cholesterol modulation, brain VLCFA increased in X-ALD mice, but decreased in controls. In murine X-ALD fibroblasts, cholesterol-lowering did not normalize VLCFA. Thus, ALDP-deficiency and VLCFA are linked to cholesterol but species differences complicate evaluating cholesterol-lowering drugs in X-ALD mice. PMID- 16213490 TI - The adiponectin paralog CORS-26 has anti-inflammatory properties and is produced by human monocytic cells. AB - The adiponectin paralog CORS-26 (collagenous repeat-containing sequence of 26kDa protein) is a member of the C1q/TNF-alpha molecular superfamily. CORS-26 is a secreted protein and baculovirus-produced CORS-26 released in the supernatant of insect cells forms stable trimers. Adiponectin exerts anti-inflammatory effects in LPS-treated monocytic cells and CORS-26 also reduces IL-6 and TNF-alpha secretion but does not increase IL-10. Suppression of NFkappaB signalling may explain the anti-inflammatory actions of CORS-26. Furthermore CORS-26 protein was detected in human monocytic and dendritic cells. The present data demonstrate for the first time that CORS-26 forms trimers, exerts anti-inflammatory properties and that it is expressed in monocytic cells. Therefore CORS-26 may provide a new target for pharmacological drugs in inflammatory diseases like the metabolic syndrome. PMID- 16213492 TI - Production of a thermostable archaeal superoxide reductase in plant cells. AB - Pyrococcus furiosus superoxide reductase (SOR) is a thermostable archaeal enzyme that reduces superoxide without producing oxygen. When produced as a fusion protein with the green fluorescent protein in plant cells, P. furiosus SOR is located in the cytosol and nucleus. The recombinant SOR enzyme retains its function and heat stability when assayed in vitro. Importantly, expressing SOR in plant cells enhances their survival at high temperature indicating that it functions in vivo. The archaeal SOR provides a novel mechanism to reduce superoxide and demonstrates the potential for using archaeal genes to alter eukaryotic metabolism. PMID- 16213493 TI - Determinants of the nucleolar targeting of protein phosphatase-1. AB - The ubiquitously expressed protein Ser/Thr phosphatase-1 isoforms PP1alpha, PP1beta and PP1gamma1 are dynamically targeted to distinct, but overlapping cellular compartments by associated proteins. Within the nucleus of HeLa cells, EGFP-tagged PP1gamma1 and PP1beta were predominantly targeted to the nucleoli, while PP1alpha showed a more diffuse distribution. Using PP1 chimaeras and point mutants we show here that a single N-terminal residue, i.e., Gln20 for PP1alpha, Arg19 for PP1beta and Arg20 for PP1gamma1 accounts for their distinct subnuclear distribution. Our data also suggest that the N-terminus of PP1beta and PP1gamma1 harbours an interaction site for one or more nucleolar interactors. PMID- 16213494 TI - Expression of macrophage-selective markers in human and rodent adipocytes. AB - CD14, CD68 and/or mouse F4/80 or human epidermal growth factor module-containing mucin-like receptor 1 (EMR1) are widely used as macrophage-specific markers. Since macrophages infiltrate several tissues during inflammatory processes, CD14, CD68 and EMR1-F4/80 have been employed to discriminate between tissue-containing macrophages, like adipose tissue (AT), and other cells. Using real-time PCR experiments, we show that isolated adipocytes from humans and mice AT express high levels of CD14 and CD68 mRNA, whereas EMR1-F4/80 is mainly present in the macrophage-containing stroma-vascular fraction. Furthermore, fibroblasts-like cells (adipoblasts), preadipocytes and adipocytes from the murine cell lines, 3T3 F442A and BFC-1, express CD14 and CD68 mRNA and protein as determined by fluorescence-activated cell sorter, but not F4/80 which, as expected, is strongly expressed in the macrophage cell line RAW264.7. These results reinforce the view that EMR1-F4/80 is the best macrophage marker to date and show that CD14 and CD68 are not macrophage-specific proteins. PMID- 16213496 TI - Caspase activation is involved in chronic periodontitis. AB - Periodontitis, a common infectious disease, is initiated by various gram-negative bacteria and characterized by the destruction of the periodontal tissue. Here, we investigated the role of caspases, intracellular proteases that are the key mediators of apoptosis. We show that activation of caspase-3 and caspase-7 is considerably enhanced in gingival tissue from patients with periodontitis. We also demonstrate in in vitro experiments that various periodontopathic bacteria exert a direct growth-suppressing effect and, moreover, can trigger a host mediated cytotoxic activity involving the CD95 death receptor. Our data suggest that caspase activation is a prominent feature in periodontitis-associated tissue injury. PMID- 16213495 TI - Evidence for proprotein convertase activity in the endoplasmic reticulum/early Golgi. AB - Processing of precursor proteins by the proprotein convertases is thought to occur mainly in the trans-Golgi network or post-Golgi compartments. Such cleavage is inhibited by the prosegment of the convertases. During our studies of the use of the inhibitory prosegment of PC1, we noticed that a construct containing the prosegment fused to the C-terminal secretory granule sorting domain was cleaved in the endoplasmic reticulum (ER) at a pair of basic residues, best recognized by furin and PC7. This was further confirmed when this construct was fused at the C terminus with a KDEL ER-retention signal. This suggests that the convertases could cleave some substrates within the ER, possibly by displacing the inhibitory prosegment associated with them. PMID- 16213497 TI - Rhinovirus-stabilizing activity of artificial VLDL-receptor variants defines a new mechanism for virus neutralization by soluble receptors. AB - Members of the low-density lipoprotein receptor family possess various numbers of ligand binding repeats that non-equally contribute to binding of minor group human rhinoviruses. Using an artificial concatemer of five copies of repeat 3 of the human very-low density lipoprotein receptor, we demonstrate protection of HRV2 against low-pH mediated uncoating and inhibition of penetration of an RNA specific fluorescent dye into the intact virion. This indicates that the recombinant receptor inhibits viral breathing and irreversible conformational modifications of the capsid that precede RNA release, providing a new mechanism for rhinovirus neutralization by soluble receptor molecules. PMID- 16213498 TI - Inhibition of matrix metalloproteinase-9 by interferons and TGF-beta1 through distinct signalings accounts for reduced monocyte invasiveness. AB - Cytokines may provide signals for regulating human monocyte matrix metalloproteinase-9 (MMP-9) activity. In this study, we investigated the roles of interferons (IFN) type I/II and transforming growth factor-beta1 (TGF-beta1) in MMP-9-mediated invasiveness. MMP-9 antibody and inhibitor, IFNs and TGF-beta1 inhibited monocyte transmigration through Matrigel. IFNs and TGF-beta1 downregulated MMP-9 mRNA, protein and activity levels. The inhibitory action of IFNs was associated with the STAT1/IRF-1 pathway since the JAK inhibitor AG490 blocked STAT1 phosphorylation, IRF-1 synthesis and counteracted the blockade of MMP-9 release. TGF-beta1-mediated MMP-9 inhibition appeared STAT1/IRF-1 independent but reversed by the protein tyrosine kinase inhibitor tyrphostin 25. Our data point out the importance of IFNs and TGF-beta1 in the control of monocyte MMP-9-mediated extravasation. PMID- 16213500 TI - How can elongation factors EF-G and EF-Tu discriminate the functional state of the ribosome using the same binding site? AB - Elongation factors EF-G and EF-Tu are structural homologues and share near identical binding sites on the ribosome, which encompass the GTPase-associated centre (GAC) and the sarcin-ricin loop (SRL). The SRL is fixed structure in the ribosome and contacts elongation factors in the vicinity of their GTP-binding site. In contrast, the GAC is mobile and we hypothesize that it interacts with the alpha helix D of the EF-Tu G-domain in the same way as with the alpha helix A of the G'-domain of EF-G. The mutual locations of these helices and GTP-binding sites in the structures of EF-Tu and EF-G are different. Thus, the orientation of the GAC relative to the SRL determines whether EF-G or EF-Tu will bind to the ribosome. PMID- 16213499 TI - Seasonal environmental changes regulate the expression of the histone variant macroH2A in an eurythermal fish. AB - Adaptation to cold and warm conditions requires dramatic change in gene expression. The acclimatization process of the common carp Cyprinus carpio L. in its natural habitat has been used to study how organisms respond to natural environmental changes. At the cellular level, adaptation to cold condition is accompanied by a dramatic alteration in nucleolar structure and a down regulation of the expression of ribosomal genes. We show that the enrichment of condensed chromatin in winter adapted cells is not correlated with an increase of the heterochromatin marker trimethyl and monomethyl K20H4. However, the expression of the tri methyl K4 H3 and of the variant histone macroH2A is significantly increased during the winter season together with a hypermethylation of CpG residues. Taking into account the properties of macroH2A toward chromatin structure and dynamics and its role in gene repression our data suggest that the increased expression of macroH2A and the hypermethylation of DNA which occurs upon winter-acclimatization plays a major role for the reorganization of chromatin structure and the regulation of gene expression during the physiological adaptation to a colder environment. PMID- 16213501 TI - A single glutamine synthetase gene produces tissue-specific subcellular localization by alternative splicing. AB - Glutamine synthetase (GS) plays a key role in two major biochemical pathways: In liver GS catalyzes ammonia detoxification, whereas in neural tissues it also functions in recycling of the neurotransmitter glutamate. In most species the GS gene gives rise to a cytoplasmic protein in both liver and neural tissues. However, in species that utilize the ureosmotic or uricotelic system for ammonia detoxification, the enzyme is cytoplasmic in neural tissues, but mitochondrial in liver cells. Since most vertebrates have a single copy of the GS gene, it is not clear how tissue-specific subcellular localization is achieved. Here we show that in the ureosmotic elasmobranch, Squalus acanthias (spiny dogfish), two different GS transcripts are generated by tissue-specific alternative splicing. The liver transcript contains an alternative exon that is not present in the neural one. This exon leads to acquisition of an upstream in-frame start codon and formation of a mitochondrial targeting signal (MTS). Therefore, the liver product is targeted to the mitochondria while the neural one is retained in the cytoplasm. These findings present a mechanism in which alternative splicing of an MTS encoding exon is used to generate tissue-specific subcellular localization. PMID- 16213502 TI - Single chain antibody against the common epitope of mutant p53 restores wild-type activity to mutant p53 protein. AB - Here, we describe the biological activity of ME1, a mouse single chain Fv fragment (scFv) against the common epitope of mutant p53, which is efficiently expressed in mammalian cells. We found that in vivo interaction of the conformational p53 mutant R175H protein with the scFv resulted in the acquisition of wild-type p53 characteristics, manifested in trans-activation of p21, as well as induction of apoptosis. Moreover, antibody binding leads to abrogation of the mutant p53 mediated "gain of function" as estimated by downregulation of EGR-1, a transcriptional target of mutant p53. These findings suggest that the scFv restores wild-type properties to mutant p53. PMID- 16213503 TI - BH3-ligand regulates access of MCL-1 to its E3 ligase. AB - A genome wide search for new BH3-containing Bcl-2 family members was conducted using position weight matrices (PWM) and identified a large (480kDa), novel BH3 only protein, originally called LASU1 (now also known as Ureb-1, E3(histone), ARF BP1, and Mule). We demonstrated that LASU1 is an E3 ligase that ubiquitinated Mcl 1 in vitro and was required for its proteasome-dependent degradation in HeLa cells. Of note, the BH3 domain of LASU1 interacted with Mcl-1 but not with Bcl-2 or Bcl-Xl. A competing BH3-ligand derived from Bim interacted with Mcl-1 and prevented its interaction with LASU1 in HeLa cells, causing elevation of the steady-state levels of Mcl-1. This suggests that the unliganded form of Mcl-1 is sensitive to LASU1-mediated degradation of Mcl-1. PMID- 16213504 TI - Characterization of estrogen receptor betab in sea bream (Sparus auratus): phylogeny, ligand-binding, and comparative analysis of expression. AB - Estrogens control many physiological processes in both female and male vertebrates, mostly mediated by specific nuclear estrogen receptors (ER). Two ER subtypes (ERalpha and ERbeta) are present in most vertebrates, including the sea bream (Sparus auratus) a hermaphrodite teleost fish. In the present study several variant cDNAs encoding a second sea bream ERbeta (sbERbetab) is reported. Phylogenetic and Southern blot analysis indicate that sbERbetab and the previously cloned sbERbetaa (formerly sbERbeta) are encoded by different genes, which may have arisen by duplication of an ancestral ERbeta gene. Competitive binding assays show that sbERbetab has high affinity for 17beta-estradiol (K(d) = 1 nM) and specifically binds estrogen agonists (diethylstilbestrol and ethynylestradiol) and antagonists (ICI 182,780). In Northern blot sbERalpha, sbERbetaa, sbERbetab produce several different transcripts in a variety of tissues. RT-PCR showed a partially overlapping but differential tissue distribution in both male and female sea bream. PMID- 16213505 TI - Telemedicine and robotics: paving the way to the globalization of surgery. AB - The concept of delivering health services at a distance, or telemedicine is becoming an emerging tool for the field of surgery. For the surgical services, telepresence surgery through robotics is gradually being incorporated into health care practices. This article will provide a brief overview of the principles surrounding telemedicine and telepresence surgery as they specifically relate to robotics. Where limitations have been reached in laparoscopy, robotics has allowed further steps forward. The development of robotics in medicine has been a progression from passive to immersive technology. In gynecology, the utilization of robotics has evolved from the use of Aesop, a robotic arm for camera manipulation, to full robotic systems such as Zeus, and the daVinci surgical system. These systems have not only been used directly for a variety of procedures but have also become a useful tool for conferencing and the mentoring of surgeons from afar. As this mode of technology becomes assimilated into the culture of surgery and medicine globally, caution must be taken to carefully navigate the economic, legal and ethical implications of telemedicine. Despite the challenges faced, telepresence surgery holds promise for more widespread applications. PMID- 16213507 TI - Subject-specific finite element models of long bones: An in vitro evaluation of the overall accuracy. AB - The determination of the mechanical stresses induced in human bones is of great importance in both research and clinical practice. Since the stresses in bones cannot be measured non-invasively in vivo, the only way to estimate them is through subject-specific finite element modelling. Several methods exist for the automatic generation of these models from CT data, but before bringing them in the clinical practice it is necessary to assess their accuracy in the predictions of the bone stresses. Particular attention should be paid to those regions, like the epiphyseal and metaphyseal parts of long bones, where the automatic methods are typically less accurate. Aim of the present study was to implement a general procedure to automatically generate subject-specific finite element models of bones from CT data and estimate the accuracy of this general procedure by applying it to one real femur. This femur was tested in vitro under five different loading scenarios and the results of these tests were used to verify how the adoption of a simplified two-material homogeneous model would change the accuracy with respect to the density-based inhomogeneous one, with special attention paid to the epiphyseal and metaphyseal proximal regions of the bone. The results showed that the density-based inhomogeneous model predicts with a very good accuracy the measured stresses (R(2)=0.91, RMSE=8.6%, peak error=27%), while the two-material model was less accurate (R(2)=0.89, RMSE=9.6%, peak error=35%). The results showed that it is possible to automatically generate accurate finite element models of bones from CT data and that the strategy of material properties mapping has a significant influence on its accuracy. PMID- 16213508 TI - Enantiomeric separation of 1-phenylethylamine and 1-cyclohexylethylamine in capillary electrophoresis with contactless conductivity detection. AB - Contactless conductivity detection was employed for the detection of the enantiomers of 1-phenylethylamine and 1-cyclohexylethylamine which were separated in capillary electrophoresis with unprecedented high resolutions R(s) of 2.3 and 3.3, respectively, by using a combination of dimethyl-beta-cyclodextrin and the chiral crown ether 18C6H4 as chiral selectors in a citric acid buffer of pH 2.4. The conductivity measurement enabled the direct detection, i.e. without having to derivatize or resort to indirect methods, of all species including the non-UV absorbing enantiomers of cyclohexylamine. Detection limits of 0.5 microM were achieved and the determination of enantiomeric ratios of up to 99:1 was found possible. PMID- 16213509 TI - Determination and characterization of organic explosives using porous graphitic carbon and liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. AB - A new LC-MS method for the determination and characterization of three groups of commonly used organic explosives (nitroaromatic compounds, cyclic nitroamines and nitrate esters) was developed using a porous graphitic carbon (PGC) (Hypercarb) column. Twenty-one different explosive-related compounds--including 2,4,6 trinitrotoluene, its by-products and its degradation products--were chromatographically separated in a single analysis. This efficient separation facilitates the identification of the manufacturer of the explosive using the identified analytes as a fingerprint. A final, conclusive identification of the analytes can be obtained using LC-MS equipped with an atmospheric pressure chemical ionization (APCI) interface. Solvent effects on chromatographic behaviour were investigated, as were the effects of solvent mixtures and mobile phase additives. The number and the relative positions of the nitro groups within analyte molecules influence their order of elution; these effects were investigated. The data thus generated can be interpreted to support a hypothesis concerning the retention mechanism of nitro-containing compounds when using PGC. Limits of detection ranged from 0.5 to 41.2 ng. The new methodology described herein improves the sensitivity and selectivity of explosive detection. The effectiveness of the method is demonstrated by the analysis of soil samples containing explosives residue from test fields in Sweden and Afghanistan. PMID- 16213510 TI - Development of a large-volume, membrane-based injection system for gradient elution micro-liquid chromatography. AB - A membrane unit which can be used to inject a large volume of sample solution was developed to facilitate reproducible and accurate gradient elution in micro-high performance liquid chromatography (micro-HPLC). Since the membrane unit has a very low void volume, it facilitates the effective concentration of analytes in large sample volumes. Gradient elution micro-HPLC with the membrane unit allowed the efficient separation of n-alkyl benzoates, used as test samples, in a short time without marked gradient delay. In this study, the membrane unit could be loaded with up to 50 microg of n-hexyl benzoate, and more than 500 microL of sample solution could be applied. In about 50 chromatographic runs, the relative standard deviation (RSD) of the relative retention time of n-hexyl benzoate with respect to methyl benzoate was 0.530%. PMID- 16213511 TI - Optimisation and robustness analysis of a hydrophobic interaction chromatography step. AB - Process development, optimisation and robustness analysis for chromatography separations are often entirely based on experimental work and generic knowledge. The present study proposes a method of gaining process knowledge and assisting in the robustness analysis and optimisation of a hydrophobic interaction chromatography step using a model-based approach. Factorial experimental design is common practice in industry today for robustness analysis. The method presented in this study can be used to find the critical parameter variations and serve as a basis for reducing the experimental work. In addition, the calibrated model obtained with this approach is used to find the optimal operating conditions for the chromatography column. The methodology consists of three consecutive steps. Firstly, screening experiments are performed using a factorial design. Secondly, a kinetic-dispersive model is calibrated using gradient elution and column load experiments. Finally, the model is used to find optimal operating conditions and a robustness analysis is conducted at the optimal point. The process studied in this work is the separation of polyclonal IgG from BSA using hydrophobic interaction chromatography. PMID- 16213512 TI - Interactions between zwitterionic and conventional anionic and cationic surfactants. AB - Critical micelle concentration (cmc) values have been determined for the mixed zwitterionic/anionic surfactant systems of N-dodecyl-N,N-dimethyl-3-ammonio-1 propanesulfonate (ZW3-12)/sodium dodecyl sulfate (SDS), N-dodecyl-N,N (dimethylammonio)butyrate (DDMAB)/SDS, N-octyl-N,N-dimethyl-3-ammonio-1 propanesulfonate (ZW3-08)/sodium octyl sulfate (SOS), and the zwitterionic/cationic systems of ZW3-12/dodecyltrimethylammonium bromide (DTAB), DDMAB/DTAB. Conductivity studies and nuclear magnetic resonance (NMR) spectroscopy were the methods employed for cmc determinations. The degree of nonideality of the interaction in the micelle (beta(m)), for each system, was determined according to Rubingh's nonideal solution theory. Evidence was found for the existence of strong interactions between zwitterionic and anionic surfactants in each of the zwitterionic/anionic systems. The ZW3-08/SOS and DDMAB/SDS systems behaved synergistically at all mole fractions studied while the ZW3-12/SDS system exhibited synergistic behavior above mole fractions of 0.30. Greater negative deviations from ideal behavior were demonstrated in the DDMAB/SDS system than in the other two zwitterionic/anionic systems. The zwitterionic/cationic systems of ZW3-12/DTAB and ZW3-08/OTAB displayed only slight deviations from ideal behavior, therefore indicating near ideal mixing. PMID- 16213514 TI - Anisotropic Brownian diffusion near a nanostructured surface. AB - The Brownian diffusion of sub-micrometer-sized particles (diameter 0.52 microm) in the vicinity of a nanostructured surface was experimentally characterized. The surface consisted of a repeating pattern of rectangular grooves with depth 35 nm and pitch 400 nm. It was found that the one-dimensional particle diffusivity parallel to the nanogrooves was significantly higher than the diffusivity perpendicular to the grooves (1.518+/-0.274 (S.E.) microm2/s compared to 0.704+/ 0.090 microm2/s). No such anisotropy in the one-dimensional Brownian diffusivity was found for particles near a flat surface. PMID- 16213513 TI - Effect of surfactant sucrose ester on physical properties of dairy whipped emulsions in relation to those of O/W interfacial layers. AB - Dairy foams were manufactured on a pilot plant with various sucrose ester contents. Oil-in-water emulsions were produced by high-pressure homogenisation of anhydrous milk fat (20 wt%) with an aqueous phase containing skim milk powder (6.5 wt%), sucrose (15 wt%), hydrocolloids (2 wt%), and sucrose esters. Sucrose ester content was varied from 0 to 0.35 wt%. Firmness and stability of dairy foams were determined. The fraction of protein associated with emulsion fat droplets and the compression isotherms of those droplets were determined as a function of sucrose ester content. With less than 0.1 wt% sucrose ester, no foam could be produced. The most firm and stable foams were obtained with ca. 0.1 wt% sucrose ester. The fraction of protein associated with emulsion droplets suddenly falls from 60% at a sucrose ester content lower than 0.1125% down to ca. 10-20% for higher surfactant content. Compression isotherms of emulsion droplets at the air-water interface show that, in the presence of surfactant, emulsion droplets disrupt and spread at the interface whilst without surfactant they become dispersed. This means that the presence of sucrose ester causes some destabilisation of fat droplet interfacial layers. There is hence an optimal sucrose ester content that allows some destabilisation of the oil-water interface without concomitant protein displacement from that interface. Consequently, with the recipe and manufacturing process used to produce dairy foams, there exists a compromise in sucrose ester content with regards to manufacture and shelf-life of dairy foams. PMID- 16213515 TI - Experimental Trypanosoma evansi infection in the goat. I. Clinical signs and clinical pathology. AB - A strain of Trypanosoma evansi isolated from an equine case of surra in Mindanao, Philippines was used to infect intravenously two groups (A and B) of five male goats aged 8-10 months. Animals of groups A and B received 5000 and 50 000 trypanosomes, respectively, and five further animals (group C) served as uninfected controls. Four of the 10 infected goats died 8-78 days after inoculation. Group C goats gained weight (mean 22.8 g/day) while infected goats in groups A and B lost weight (means of 21.4 and 45.0 g/day, respectively). Parasitaemia fluctuated regularly between peaks and troughs, with repeated periods of about 6 days during which no trypanosomes were detected in the blood. Clinical signs and clinico-pathological changes in infected goats were not pathognomonic in the absence of parasites in the blood, and leucocytosis was not a reliable indicator of infection. It was concluded that in endemic areas fluctuating fever, progressive emaciation, anaemia, coughing, testicular enlargement and diarrhoea are suggestive of surra; confirmation, however, may necessitate examination of blood every few days for trypanosomes, and possibly other diagnostic tests. PMID- 16213517 TI - Immunohistochemical diagnosis of alimentary lymphomas and severe intestinal inflammation in cats. AB - Intestinal tissue samples were examined from 32 cats in which a histopathological diagnosis of alimentary lymphoma or multicentric lymphoma affecting the gastrointestinal tract had been made. These samples were re-evaluated histopathologically and serial sections were examined immunohistochemically with antisera specific for the lymphoid markers CD3, CD79a and BLA-36 and for class II molecules of the major histocompatability complex. The cats ranged in age from 4 16 years (median 10.5 years). The main presenting clinical signs were vomiting, diarrhoea and weight loss. The majority of alimentary lymphomas were of the B cell type (n=15), whereas cases of T-cell lymphoma were fewer in number (n=8). Four cats had lymphoma of a mixed T-and B-cell phenotype. In five of the cats, immunohistochemistry suggested an inflammatory process, in contradiction to the original histopathological diagnosis of lymphoma. Immunolabelling would appear to be a useful adjunct to histopathology in classifying cases of feline alimentary lymphoma, and may help in distinguishing lymphoma from severe intestinal inflammation. PMID- 16213518 TI - Roles of double-strand breaks, nicks, and gaps in stimulating deletions of CTG.CAG repeats by intramolecular DNA repair. AB - A series of plasmids harboring CTG.CAG repeats with double-strand breaks (DSB), single-strand nicks, or single-strand gaps (15 or 30 nucleotides) within the repeat regions were used to determine their capacity to induce genetic instabilities. These plasmids were introduced into Escherichia coli in the presence of a second plasmid containing a sequence that could support homologous recombination repair between the two plasmids. The transfer of a point mutation from the second to the first plasmid was used to monitor homologous recombination (gene conversion). Only DSBs increased the overall genetic instability. This instability took place by intramolecular repair, which was not dependent on RuvA. Double-strand break-induced instabilities were partially stabilized by a mutation in recF. Gaps of 30 nt formed a distinct 30 nt deletion product, whereas single strand nicks and gaps of 15 nt did not induce expansions or deletions. Formation of this deletion product required the CTG.CAG repeats to be present in the single stranded region and was stimulated by E.coli DNA ligase, but was not dependent upon the RecFOR pathway. Models are presented to explain the intramolecular repair-induced instabilities and the formation of the 30 nt deletion product. PMID- 16213516 TI - Experimental Trypanosoma evansi infection in the goat. II. Pathology. AB - Infection of male goats aged 8-10 months with 5000 or 50 000 organisms of a Mindanao strain of Trypanosoma evansi was observed over a period of 90 days. The infection induced clinical disease which was lethal, especially at the higher dose rate. Lesions were more acute in goats that received the higher dose. Gross and microscopical changes were not pathognomonic, except in the presence of demonstrable trypanosomes. At necropsy, a combination of lymphadenopathy, splenomegaly, hepatomegaly, testicular enlargement, anaemic signs and consolidation of the anterior lobes of the lungs was suggestive of surra. Testicular changes, especially aspermia, indicated probable infertility. The cytopathology of the lungs, liver, intestine, kidneys, testes, bone marrow, brain and other organs was immunological in nature, characterized by mononuclear infiltration of interstitial tissues, with minor cellular damage and the presence of trypanosomes. B- and T- cell responses were observed in the lymphatic system, but the findings indicated immunosuppression in the lymph nodes, spleen and bone marrow during the third month after infection. Exudative inflammatory changes were mild. It is suggested that the cytopathology of most haemophilic trypanosomal infections is predominantly an immunological process. PMID- 16213519 TI - Preprotein translocase of the outer mitochondrial membrane: reconstituted Tom40 forms a characteristic TOM pore. AB - Tom40 is the central pore-forming component of the translocase of the outer mitochondrial membrane (TOM complex). Different views exist about the secondary structure and electrophysiological characteristics of Tom40 from Saccharomyces cerevisiae and Neurospora crassa. We have directly compared expressed and renatured Tom40 from both species and find a high content of beta-structure in circular dichroism measurements in agreement with refined secondary structure predictions. The electrophysiological characterization of renatured Tom40 reveals the same characteristics as the purified TOM complex or mitochondrial outer membrane vesicles, with two exceptions. The total conductance of the TOM complex and outer membrane vesicles is twofold higher than the total conductance of renatured Tom40, consistent with the presence of two TOM pores. TOM complex and outer membrane vesicles possess a strongly enhanced sensitivity to a mitochondrial presequence compared to Tom40 alone, in agreement with the presence of several presequence binding sites in the TOM complex, suggesting a role of the non-channel Tom proteins in regulating channel activity. PMID- 16213520 TI - A GAF domain in the hypoxia/NO-inducible Mycobacterium tuberculosis DosS protein binds haem. AB - The majority of the Mycobacterium tuberculosis response to hypoxia and nitric oxide is through the DosRS (DevRS) two-component regulatory system. The N terminal input domain of the DosS sensor contains two GAF domains. We demonstrate here that the proximal GAF domain binds haem, and identified histidine 149 of DosS as critical to haem-binding; the location of this histidine residue is similar to the cGMP-binding site in a crystal structure of cyclic nucleotide phosphodiesterase 2A. GAF domains are frequently involved in binding cyclic nucleotides, but this is the first GAF domain to be identified that binds haem. In contrast, PAS domains (similar to GAF domains in structure but not primary sequence) frequently use haem cofactors, and these findings further illustrate how the functions of these domains overlap. We propose that the activation of the DosS sensor is controlled through the haem binding of molecular oxygen or nitric oxide. PMID- 16213521 TI - Cofilin increases the torsional flexibility and dynamics of actin filaments. AB - We have measured the effects of cofilin on the conformation and dynamics of actin filaments labeled at Cys374 with erythrosin-iodoacetemide (ErIA), using time resolved phosphorescence anisotropy (TPA). Cofilin quenches the phosphorescence intensity of actin-bound ErIA, indicating that binding changes the local environment of the probe. The cofilin concentration-dependence of the phosphorescence intensity is sigmoidal, consistent with cooperative actin filament binding. Model-independent analysis of the anisotropies indicates that cofilin increases the rates of the microsecond rotational motions of actin. In contrast to the reduction in phosphorescence intensity, the changes in the rates of rotational motions display non-nearest-neighbor cooperative interactions and saturate at substoichiometric cofilin binding densities. Detailed analysis of the TPA decays indicates that cofilin decreases the torsional rigidity (C) of actin, increasing the thermally driven root-mean-square torsional angle between adjacent filament subunits from approximately 4 degrees (C = 2.30 x 10(-27) Nm2 radian( 1)) to approximately 17 degrees (C = 0.13 x 10(-27) Nm2 radian(-1)) at 25 degrees C. We favor a mechanism in which cofilin binding shifts the equilibrium between thermal ErIA-actin filament conformers, and facilitates two distinct structural changes in actin. One is local in nature, which affects the structure of actin's C terminus and is likely to mediate nearest-neighbor cooperative binding and filament severing. The second is a change in the internal dynamics of actin, which displays non-nearest-neighbor cooperativity and increases the torsional flexibility of filaments. The long-range effects of cofilin on the torsional dynamics of actin may accelerate P(i) release from filaments and modulate interactions with other regulatory actin filament binding proteins. PMID- 16213523 TI - The biosynthesis of mycolic acids in Mycobacterium tuberculosis relies on multiple specialized elongation complexes interconnected by specific protein protein interactions. AB - Tuberculosis kills about two million people every year and remains one of the leading causes of mortality worldwide. As a result of the increasing antibiotic resistance of Mycobacterium tuberculosis (Mtb) strains, there is an urgent need for new antitubercular drugs. Several efficient antibiotics, including isoniazid, specifically target the fatty acid synthase-II (FAS-II) complex of mycolic acid biosynthesis. We have previously shown that there are protein-protein interactions between the components of FAS-II that are essential for mycobacterial survival. We have now looked at the potential partners of FAS-II, mtFabD, the methyltransferases MmaAs, and Pks13. A combination of yeast two hybrid and co-immunoprecipitation experiments showed that mtFabD interacts with each beta-ketoacyl-synthase (KasA, KasB and mtFabH) and with the core of FAS-II (InhA and MabA). The methyltransferases have a greater affinity for KasA and KasB than for mtFabH, suggesting that modifications on the meromycolic chains may occur during their elongation. Finally, Pks13, which catalyzes the final Claisen condensation of mycolic acids, interacts specifically with KasB. These data allowed us to determine the architecture of the multiple specialized FAS-II complexes, giving us insights into the organization of the complete mycolic acids biosynthesis. Our studies suggest a new and crucial interaction (KasB-Pks13) as a putative target for peptidomimetic antibiotics. PMID- 16213522 TI - The structure of the C5a receptor-blocking domain of chemotaxis inhibitory protein of Staphylococcus aureus is related to a group of immune evasive molecules. AB - The chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) is a 121 residue excreted virulence factor. It acts by binding the C5a- (C5aR) and formylated peptide receptor (FPR) and thereby blocks specific phagocyte responses. Here, we report the solution structure of a CHIPS fragment consisting of residues 31-121 (CHIPS31-121). CHIPS31-121 has the same activity in blocking the C5aR compared to full-length CHIPS, but completely lacks FPR antagonism. CHIPS31-121 has a compact fold comprising an alpha-helix (residues 38-51) packed onto a four-stranded anti-parallel beta-sheet. Strands beta2 and beta3 are joined by a long loop with a relatively well-defined conformation. Comparison of CHIPS31 121 with known structures reveals striking homology with the C-terminal domain of staphylococcal superantigen-like proteins (SSLs) 5 and 7, and the staphyloccocal and streptococcal superantigens TSST-1 and SPE-C. Also, the recently reported structures of several domains of the staphylococcal extracellullar adherence protein (EAP) show a high degree of structural similarity with CHIPS. Most of the conserved residues in CHIPS and its structural homologues are present in the alpha-helix. A conserved arginine residue (R46 in CHIPS) appears to be involved in preservation of the structure. Site-directed mutagenesis of all positively charged residues in CHIPS31-121 reveals a major involvement of arginine 44 and lysine 95 in C5aR antagonism. The structure of CHIPS31-121 will be vital in the further unraveling of its precise mechanism of action. Its structural homology to S.aureus SSLs, superantigens, and EAP might help the design of future experiments towards an understanding of the relationship between structure and function of these proteins. PMID- 16213524 TI - Molecular dynamics simulations of Alzheimer's beta-amyloid protofilaments. AB - Filamentous amyloid aggregates are central to the pathology of Alzheimer's disease. We use all-atom molecular dynamics (MD) simulations with explicit solvent and multiple force fields to probe the structural stability and the conformational dynamics of several models of Alzheimer's beta-amyloid fibril structures, for both wild-type and mutated amino acid sequences. The structural models are based on recent solid state NMR data. In these models, the peptides form in-register parallel beta-sheets along the fibril axis, with dimers of two U shaped peptides located in layers normal to the fibril axis. Four different topologies are explored for stacking the beta-strand regions against each other to form a hydrophobic core. Our MD results suggest that all four NMR-based models are structurally stable, and we find good agreement with dihedral angles estimated from solid-state NMR experiments. Asp23 and Lys28 form buried salt bridges, resulting in an alternating arrangement of the negatively and positively charged residues along the fibril axis that is reminiscent of a one-dimensional ionic crystal. Interior water molecules are solvating the buried salt-bridges. Based on data from NMR measurements and MD simulations of short amyloid fibrils, we constructed structural models of long fibrils. Calculated X-ray fiber diffraction patterns show the characteristics of packed beta-sheets seen in experiments, and suggest new experiments that could discriminate between various fibril topologies. PMID- 16213526 TI - The role of sales of new motorcycles in a recent increase in motorcycle mortality rates. AB - INTRODUCTION: The National Highway Traffic Safety Administration (NHTSA) has reported that mortality rates from crashes among motorcycle riders in the United States increased from 21.0 per 100 million motorcycle miles traveled in 1997 to 38.4 per 100 million motorcycle miles traveled in 2003. At the same time, annual domestic sales of new, on-road motorcycles increased from 247,000 in 1997 to 648,000 in 2003. METHOD: This study used data from the NHTSA Fatality Analysis Reporting System and annual sales figures for on-road motorcycles to determine if newer motorcycles were more likely to be involved in fatal crashes and if fatal crashes involving newer motorcycles could account for the mortality increase after 1997. RESULTS: Mortality rates were 7.9, 8.1, 5.4, and 2.9 per 10,000 motorcycles sold for motorcycles <1, 1-3, 4-6, and 7-11 years old, respectively, from 1994 to 2003. Assuming complete registration, the number of motorcycles sold during the 2000-2003 time period accounted for 42.4% of the total number of motorcycles registered in 2003. Motorcycles sold during 2000-2003 were associated with 52.5% of all motorcycle deaths in 2003. The increase in the number of deaths associated with motorcycles less than four years old between 1997 and 2003 accounted for 78.1% of the total increase in motorcyclist deaths over this time period. CONCLUSIONS: Two possible explanations for the association between high sales volumes and mortality rates are: (a) increased exposure from more extensive use of motorcycles when they are new; and (b) inexperience with motorcycle riding or with specific motorcycles. IMPACT ON INDUSTRY: This study suggests that the deaths of growing numbers of motorcyclists are a consequence of the financial success of the motorcycle industry. PMID- 16213525 TI - Androgen receptor binding sites identified by a GREF_GATA model. AB - Changes in transcriptional regulation can be permissive for tumor progression by allowing for selective growth advantage of tumor cells. Tumor suppressors can effectively inhibit this process. The PMEPA1 gene, a potent inhibitor of prostate cancer cell growth is an androgen-regulated gene. We addressed the question of whether or not androgen receptor can directly bind to specific PMEPA1 promoter upstream sequences. To test this hypothesis we extended in silico prediction of androgen receptor binding sites by a modeling approach and verified the actual binding by in vivo chromatin immunoprecipitation assay. Promoter upstream sequences of highly androgen-inducible genes were examined from microarray data of prostate cancer cells for transcription factor binding sites (TFBSs). Results were analyzed to formulate a model for the description of specific androgen receptor binding site context in these sequences. In silico analysis and subsequent experimental verification of the selected sequences suggested that a model that combined a GREF and a GATA TFBS was sufficient for predicting a class of functional androgen receptor binding sites. The GREF matrix family represents androgen receptor, glucocorticoid receptor and progesterone receptor binding sites and the GATA matrix family represents GATA binding protein 1-6 binding sites. We assessed the regulatory sequences of the PMEPA1 gene by comparing our model-based GREF_GATA predictions to weight matrix-based predictions. Androgen receptor binding to predicted promoter upstream sequences of the PMEPA1 gene was confirmed by chromatin immunoprecipitation assay. Our results suggested that androgen receptor binding to cognate elements was consistent with the GREF_GATA model. In contrast, using only single GREF weight matrices resulted in additional matches, apparently false positives. Our findings indicate that complex models based on datasets selected by biological function can be superior predictors as they recognize TFBSs in their functional context. PMID- 16213527 TI - Detection of 14-3-3 protein in the cerebrospinal fluid in mitochondrial encephalopathy with lactic acidosis and stroke-like episodes. AB - We describe a 13-year-old boy with mitochondrial encephalopathy with lactic acidosis and stroke-like episodes (MELAS) who experienced a stroke-like episode resulting in severe mental regression and quadriplegia. We tested 14-3-3 protein in the cerebrospinal fluid (CSF) of the patient four times around a stroke-like episode in a magnetic resonance imaging (MRI) study. Detection of the protein in the CSF was well correlated with the clinical course and range of damage of the brain lesion on MRI. Interestingly, 14-3-3 CSF protein was detected at the beginning of mitochondrial encephalopathy without new MRI abnormalities, suggesting that it is a sensitive brain marker. We conclude that 14-3-3 CSF protein is a useful biological marker of brain disruption in MELAS as well as other neurological disorders. PMID- 16213528 TI - A Western Australian kindred with Dutch cerebral amyloid angiopathy. AB - A family from the south of Western Australia is described with Dutch cerebral amyloid angiopathy (HCHWA-D). The proband died at age 60 from recurrent lobar haemorrhages in the brain, as did his sister and five other family members. The APP 693 mutation at position 22 of the Abetapeptide resulting in a glutamine for glutamic acid was identified in the proband and the affected sister. Pathologically lobar haemorrhages were found with cerebrovascular angiopathy; neuritic plaques were found but no neurofibrilary tangles. There was a leukoencephalopathy on MRI scanning. Dementia and cognitive decline has not been observed in this family. This is the first family reported outside of Europe and the Northern Hemisphere. The discovery highlights the importance of detecting this rare cause of fatal cerebral haemorrhage as it has implications for gene testing and general medical management. PMID- 16213529 TI - Individual discrimination capability and collective decision-making. AB - Amplification is the main component of many collective phenomena in social and gregarious insects. In a society, individuals face a mixed palette of odours coming from different groups (lines, strains) and individuals present discrimination capabilities. However, often at the collective level, different groups may cooperate and act together. To understand this apparent contradiction, we use a model of food recruitment where each group of foragers have its own blend of pheromone trail that is partly recognized by the others groups. The model shows that a low level of recognition between signals is sufficient to produce a collaborative pattern between groups and that beyond a critical value of recognition, only the aggregation of all the groups around the same food source is observed. The comparison between this model and one describing the site selection by gregarious insects (e.g. cockroach) suggests that such collective response is a generic property of social phenomena governed by amplification processes. PMID- 16213530 TI - Apparent diffusion coefficient in the aging mouse brain: a magnetic resonance imaging study. AB - Novel magnetic resonance imaging sequences have and still continue to play an increasing role in neuroimaging and neuroscience. Among these techniques, diffusion-weighted imaging (DWI) has revolutionized the diagnosis and management of diseases such as stroke, neoplastic disease and inflammation. However, the effects of aging on diffusion are yet to be determined. To establish reference values for future experimental mouse studies we tested the hypothesis that absolute apparent diffusion coefficients (ADC) of the normal brain change with age. A total of 41 healthy mice were examined by T2-weighted imaging and DWI. For each animal ADC frequency histograms (i) of the whole brain were calculated on a voxel-by-voxel basis and region-of-interest (ROI) measurements (ii) performed and related to the animals' age. The mean entire brain ADC of mice <3 months was 0.715(+/-0.016) x 10(-3) mm2/s, no significant difference to mice aged 4 to 5 months (0.736(+/-0.040) x 10(-3) mm2/s) or animals older than 9 months 0.736(+/ 0.020) x 10(-3) mm2/s. Mean whole brain ADCs showed a trend towards lower values with aging but both methods (i + ii) did not reveal a significant correlation with age. ROI measurements in predefined areas: 0.723(+/-0.057) x 10(-3) mm2/s in the parietal lobe, 0.659(+/-0.037) x 10(-3) mm2/s in the striatum and 0.679(+/ 0.056) x 10(-3) mm2/s in the temporal lobe. With advancing age, we observed minimal diffusion changes in the whole mouse brain as well as in three ROIs by determination of ADCs. According to our data ADCs remain nearly constant during the aging process of the brain with a small but statistically non-significant trend towards a decreased diffusion in older animals. PMID- 16213531 TI - Angiotensin II type 1 (AT1)-receptor blocker prevents impairment of endothelium dependent cerebral vasodilation by acute cigarette smoking in rats. AB - Our aim was to test for smoking-induced endothelial dysfunction in rat cerebral vessels, then to evaluate the effect of valsartan [angiotensin II type I (AT1) receptor blocker] on that impairment. In pentobarbital-anesthetized, mechanically ventilated Sprague-Dawley rats, we used a cranial window preparation to measure changes in pial vessel diameters following topical applications of acetylcholine (Ach) (before and after smoking or intravenous nicotine infusion; n = 6 in each group), and adenosine (n = 6 for before and after smoking). Then, after intravenous valsartan pretreatment we reexamined the pial vasodilator response to topical Ach (before and after cigarette smoking). Under control conditions, cerebral arterioles were dilated by 6.9 +/- 4.2% and 13.6 +/- 4.8% by topical Ach (10(-6) M and 10(-5) M, respectively) and by 6.4 +/- 2.5% and 12.2 +/- 3.1% by topical adenosine (10(-5) M and 10(-4) M, respectively). One hour after a 1-min inhalation of mainstream smoke (1-mg nicotine cigarette), 10(-5) M Ach constricted cerebral arterioles (-4.4 +/- 4.1%), while 10(-4) M adenosine dilated them by 13.4 +/- 3.4%. One hour after a 1-min nicotine infusion (0.05 mg), 10(-5) M Ach dilated cerebral arterioles by 9.9 +/- 2.4%. Thus, vasodilator response to topical Ach was impaired after smoking, whereas that to adenosine was unaffected. However, the vasodilator response to Ach was unaffected by intravenous nicotine. Valsartan prevented smoking from impairing Ach-induced vasodilation. In conclusion, acute single-cigarette smoking causes a dysfunction of endothelium dependent, but not endothelium-independent, vasodilation of rat cerebral vessels in vivo, and the effect was not mimicked by intravenous nicotine. AT1-receptor blockade prevented the above smoking-induced impairment of endothelium-dependent vasodilation. PMID- 16213532 TI - Screening the receptorome for plant-based psychoactive compounds. AB - Throughout time, humans have used psychoactive plants and plant-derived products for spiritual, therapeutic and recreational purposes. Furthermore, the investigation of psychoactive plants such as Cannabis sativa (marijuana), Nicotiana tabacum (tobacco) and analogues of psychoactive plant derivatives such as lysergic acid diethylamide (LSD) have provided insight into our understanding of neurochemical processes and diseases of the CNS. Currently, many of these compounds are being used to treat a variety of diseases, such as depression and anxiety in the case of Piper methysticum Kava Kava (Martin et al., 2002; Singh and Singh, 2002). G-protein coupled receptors (GPCRs) are the most common molecular target for both psychoactive drugs and pharmaceuticals. The "receptorome" (that portion of the genome encoding ligand reception) encompasses more than 8% of the human genome (Roth et al., 2004) and as such provides a large number of possible targets for psychoactive drug interactions. A systematic, comprehensive study is necessary to identify novel active psychoactive plant based compounds and the molecular targets of known compounds. Herein we describe the development of a high throughput system (HTS) to screen psychoactive compounds against the receptorome and present two examples (Salvia divinorum, the "magic mint" hallucinogen and Banisteriopsis caapi, the main component of Ayahuasca, a psychoactive beverage) where HTS enabled the identification of the molecular target of each compound. PMID- 16213534 TI - Partial colony mortality reflects coral community dynamics: a fringing reef study near a small river in Okinawa, Japan. AB - The relative performance of (i) percent live cover, (ii) colony density, (iii) generic richness, (iv) partial colony mortality, and (v) colony size of hard corals were evaluated to determine which variables best discriminated the coral communities near a small river in Okinawa, Japan. An analysis of their variance was undertaken across a combination of sites at three depths, at increasing distance from a river's influence (Zatsun River). The river provides a periodic and localized input of sediment and fresh water to the adjacent coral reef; the effects of which we assume attenuate at increasing distance from the river mouth. The mean frequency of partially dead coral colonies (i.e., the proportion of live coral colonies that were partially dead) presented the clearest and most reliable response to river affect, and the power to discriminate among sites improved steadily with increasing depth. Spatial examination of the prevalence of partial mortality, regardless of how long ago the infliction occurred, provides a clear window to long-term processes involving population and community change and indeed the reef building capacity of the communities. PMID- 16213533 TI - Psychopharmacology of the hallucinogenic sage Salvia divinorum. AB - At present, the Mexican mint Salvia divinorum is an unregulated hallucinogen. This has resulted in various on-line botanical companies advertising and selling S. divinorum as a legal alternative to other regulated plant hallucinogens. It is predictable that its misuse will increase rapidly. The active ingredient in S. divinorum is the neoclerodane diterpene, salvinorin A (1a), which has been shown to be a kappa agonist both in vitro and in vivo. This review will cover the current state of research into the psychopharmacology of S. divinorum. PMID- 16213535 TI - Enhanced nicotinic acetylcholine receptor-mediated [3H]norepinephrine release from neonatal rat hypothalamus. AB - Nicotinic acetylcholine receptor (nAChR)-evoked release of norepinephrine (NE) has been demonstrated in a number of brain regions that receive sole noradrenergic innervation from the locus coeruleus (LC). Many of these structures display enhanced nicotine-stimulated NE release in the neonate. We have examined the hypothalamus in order to determine if this region, which receives NE projections from both the LC and medullary catecholaminergic nuclei, also demonstrates maturational changes in nAChR-mediated NE release. Quantification of radiolabeled-NE release from rat hypothalamus slices by a maximally effective dose of nicotine revealed a peak response during the first postnatal week. This was followed by a decrease at postnatal day (P) 14, and a second peak at P21. Thereafter, release was equivalent to that observed at P14. Comparison of the pharmacological properties of nAChRs mediating NE release in neonatal (P7) and mature hypothalamus suggested involvement of different nAChR subtypes at the two ages. Using the selective toxin, DSP-4, nAChR-mediated NE release in the neonatal hypothalamus was shown to be from LC terminals. Our findings demonstrate an early sensitivity of hypothalamic LC terminals to nAChR regulation that may be associated with development of systems controlling critical homeostatic functions such as stress, feeding and cardiovascular regulation. PMID- 16213536 TI - 4-benzyl-3-phenyl-5H-furan-2-one, a vasodilator isolated from Malbranchea filamentosa IFM 41300. AB - Screening of Malbranchea filamentosa IFM 41300 for bioactive compounds led to the identification of 4-benzyl-3-phenyl-5H-furan-2-one (1) as a vasodilator and erythroglaucin (2). The structure of 1 was established on the basis of spectroscopic and chemical investigations. Compound 1 inhibited Ca2+-induced vasocintraction in aortic rings pretreated with high K+ (60mM) or norepinephrine. Finally, compound 1 did not exhibit activity against human pathogenic microorganisms. PMID- 16213537 TI - Attenuation coefficient estimation using experimental diffraction corrections with multiple interface reflections. AB - The ultrasonic attenuation coefficient of a fluid or solid is an acoustic parameter routinely estimated for the purpose of materials characterization and defect/disease detection. This paper describes a broadband attenuation coefficient estimation technique that combines two established estimation approaches. The key elements of these two approaches are: (1) the use of magnitude spectrum ratios of front surface, first back surface, and second back surface reflections from interfaces of materials with plate-like geometries, and (2) the use of an experimental diffraction correction approach to avoid diffraction losses. The combined estimation approach simplifies the attenuation coefficient estimation process by eliminating the need to explicitly make diffraction corrections or calculate reflection/transmission coefficients. The approach yields estimates of the attenuation coefficient, reflection coefficient, and material density. Models, experimental procedures, and signal analysis procedures, which support implementation of the approach, are presented. Attenuation coefficient and reflection coefficient estimates are presented for water and solid samples with estimates based on measurements made with multiple transducers. PMID- 16213538 TI - Studies of a novel sensor for assessing the spatial distribution of cavitation activity within ultrasonic cleaning vessels. AB - This paper describes investigations of the spatial distribution of cavitation activity generated within an ultrasonic cleaning vessel, undertaken using a novel cavitation sensor concept. The new sensor monitors high frequency acoustic emissions (>1 MHz) generated by micron-sized bubbles driven into acoustic cavitation by the applied acoustic field. Novel design features of the sensor, including its hollow, cylindrical shape, provide the sensor with spatial resolution, enabling it to associate the megahertz acoustic emissions produced by the cavitating bubbles with specific regions of space within the vessel. The performance of the new sensor has been tested using a 40 kHz ultrasonic cleaner employing four transducers and operating at a nominal electrical power of 140 W under controlled conditions. The results demonstrate the ability of the sensors to identify 'hot-spots' and 'cold-spots' in cavitation activity within the vessel, and show good qualitative agreement with an assessment of the spatial distribution of cavitation determined through erosion monitoring of thin sheets of aluminium foil. The implications of the studies for the development of reliable methods of quantifying the performance of cleaning vessels are discussed in detail. PMID- 16213539 TI - Vibration of piezoelectric elements surrounded by fluid media. AB - In this paper we analyse vibrational characteristics of piezoceramic shells surrounded by acoustic media. Main results are presented for radially polarized piezoceramic PZT5 elements of hollow cylindrical shapes. The coupling in the radial direction between the solid and the acoustic media is accounted for indirectly, via impedance boundary conditions. The model based on such impedance boundary condition approximations offers a robust simplified alternative to a full scale fluid-solid interaction modelling. By using this model, we analyse numerically the influence of the boundary conditions imposed in the axial direction for long, medium, and short (disk-like) piezoceramic elements. PMID- 16213540 TI - Thermal fluorescence quenching properties of dissolved organic matter. AB - The fluorescence excitation-emission matrices of dissolved organic matter (DOM) are investigated between 10 and 45 degrees C for river and waste waters and organic matter standards. With increased temperature, fluorescence intensity is quenched. It is demonstrated that for a range of river and wastewater samples, that tryptophan-like fluorescence exhibits a greater range of quenching (between 20+/-4% and 35+/-5%) than fulvic-like fluorescence (19+/-4 to 26+/-3%) over this temperature range. Humic substance standards exhibit similar fulvic-like (23+/ 4%) fluorescence thermal quenching properties to river water samples (23+/-3%); however none of the samples exhibit quenching of tryptophan-like fluorescence to the same extent as the tryptophan standards (approximately 50%). Thermal fluorescence quenching is related to the exposure of the fluorophores to the heat source; our findings suggest that the tryptophan-like groups within DOM is more exposed in untreated wastewaters than in treated wastewaters riverine DOM. Thermal fluorescence properties have the potential to be used to source DOM, to provide additional chemical structural information, to temperature correct laser induced remotely sensed DOM fluorescence, and to characterise DOM through the wastewater treatment process. PMID- 16213541 TI - Short-term harmful effects of ammonia nitrogen on activated sludge microfauna. AB - The response of activated sludge microfauna in terms of abundance and diversity has been analysed to evaluate both the toxic effect of ammonia nitrogen and the acclimatisation capacity of these microorganisms to its toxicity. The harmful effect of ammonia nitrogen was studied by means of two toxicological tests. The ammonia concentrations tested were: 9, 20, 30 and 50mg NH4+-N/l in the first toxicological test and 30, 40, 50 and 80 mg NH4+-N/l in the second. The results suggest that ammonia nitrogen causes a clear but reversible toxic effect on microfauna abundance when its concentrations are around three times higher than that which the microfauna is used to. Chilodonella uncinata and Acineria uncinata were the ciliates least affected by the ammonia nitrogen toxicity. Furthermore, the majority of microfauna groups analysed (gymnamoebae and ciliates) showed capability for acclimatisation to ammonia nitrogen in terms of abundance. PMID- 16213542 TI - Kinetics of the chemical oxidation of polysulfide anions in aqueous solution. AB - The kinetic properties of the chemical oxidation of aqueous polysulfide solutions have been studied in phosphate-buffered systems at pH 7-12, at temperatures between 20 and 40 degrees C, and ionic strength between 0.05 and 0.50 M. Polysulfide solutions were mixed with a buffer solution of known dissolved oxygen concentration, after which the decrease in the oxygen concentration of the solution was measured in time. The rate of oxygen consumption can be described by the empirical relation d[O2]/dt= -k[Sx(2-)][O2](0.59) . The reaction rate constant k is moderately dependent on pH and goes through a maximum at pH 10. The rate of oxygen consumption for polysulfide solutions is approximately four times higher than for sulfide solutions. At pH values below 9, reaction products were formed according to Sx(2-)+3/2O2-->S2O3(2-)+(x-2)S(0) . At pH values higher than 9, more thiosulfate and additional sulfide were formed, which is attributed to the low chemical stability of the sulfur of oxidation state zero, formed upon polysulfide oxidation. Our results strongly suggest that hydrolysis of this 'nascent' elemental sulfur to HS- and S2O3(2-) is faster than hydrolysis of crystalline inorganic sulfur or colloidal particles of biologically produced sulfur, and has a significant contribution already at 30 degrees C and pH 10. PMID- 16213543 TI - Inhibitors designed for the active site of dihydroorotase. AB - Four new compounds have been synthesized as potential inhibitors of dihydroorotase from Escherichia coli. NMR spectroscopy was used to show that 4,6 dioxo-piperidine-2(S)-carboxylic acid (3), exists in solution as a mixture of the hydrate (7), enol (8), and enolate (9) tautomeric forms. This compound was found to be a competitive inhibitor versus dihydroorotate and thio-dihydroorotate at pH values of 7-9. The K(i) of 76 microM was lowest at pH7.0 where the ketone and hydrate forms of the inhibitor 3 predominate in solution. Compound 3 was reduced to the two diastereomeric 4-hydroxy derivatives (4 and 5) and then dehydrated to yield the alkene derivative, 1,2,3,6-tetrahydro-6-oxopyridine-2(S)-carboxylic acid (6). Compounds 4-6 were competitive inhibitors versus thio-dihydroorotate at pH 8.0 with K(i) values of 3.0, 1.6, and 2.3 mM. Dihydroorotase was unable to dehydrate the 4-hydroxy derivative 4 or 5 to the alkene 6 or catalyze the reverse reaction. PMID- 16213544 TI - Determination of total arsenic content in water by atomic absorption spectroscopy (AAS) using vapour generation assembly (VGA). AB - Analysis of arsenic in water is important in view of contamination of ground water with arsenic in some parts of the world including West Bengal in India and neighboring country Bangladesh. WHO has fixed the threshold for arsenic in drinking water to 10ppb (microg/l) level, hence the methodology for determination of arsenic is required to be sensitive at ppb level. Atomic absorption spectrophotometry with vapour generation assembly (AAS-VGA) is well known technique for the trace analysis of arsenic. However, total arsenic analysis [As(III)+As(V)] is very crucial and it requires reduction of As(V) to As(III) for correct analysis. As(III) is reduced to AsH3 vapours and finally to free As atoms, which are responsible for absorption signal in AAS. To accomplish this the vapour generation assembly attached to AAS has acid channel filled with 10 M HCl and the reduction channel with sodium borohydride. Further sample can be reduced either before aspiration for analysis, using potassium iodide (KI) or the sample can be introduced in the instrument directly and KI can be added in the reduction channel along with the sodium borohydride. The present work shows that samples prepared in 3 M HCl can be reduced with KI for 30 min before introduction in the instrument. Alternatively samples can be prepared in 6 M HCl and directly aspirated in AAS using KI in VGA reduction channel. The latter methodology is more useful when the sample size is large and time cycle is difficult to maintain. It is observed that the acid concentration of the sample in both the situations plays an important role. Further reduction in acid concentration and analysis time is achieved for the arsenic analysis by using modified method. Analysis in both the methods is sensitive at ppb level. PMID- 16213545 TI - Calcite-seeded crystallization of calcium phosphate for phosphorus recovery. AB - For phosphorus recovery from wastewater, the present paper aims at understanding the crystallization of calcium phosphate by using calcites (Juraperle and Coccolith) as seeds from hard water. Synthetic hard water with 60 mgCl(-1) carbonate and 10 mgPl(-1) phosphate was prepared and used in the batch experiments of calcite-seeded crystallization. The solution composition was measured throughout the batch crystallization process, and the corresponding saturation indices with respect to the minerals were calculated with PHREEQC Program. The surface of the calcite seeds was observed and measured with environmental scanning electron microscopy (ESEM), Fourier transform infrared (FTIR) and BET method. The studies show that both calcites are effective seeds for the crystallization of calcium phosphate from hard water; the used calcites are more efficient than the original ones because newly formed crystals with calcium phosphate have covered their surfaces. The studies show that the calcite seeds can be used for phosphorus recovery from hard waters. PMID- 16213547 TI - Experimental study on the thermal oxidation of 2-chlorophenol in air over the temperature range 450-900 degrees C. AB - The thermal oxidation of 2-chlorophenol (2-CP) in air was investigated using a perfectly stirred reactor at 1 atm over the temperature range 450-900 degrees C. The relative concentration of 2-CP was 1,000 ppm V (equivalence ratio Phi=0.03). About fifty organic products were identified as trace species. The concentration profiles of 2-CP, carbon oxides as well as those of seventeen major organic intermediates and six non-to-lower chlorinated dioxins and furans were presented as a function of temperature for a residence time of 2s. The most abundant intermediate products were carbon monoxide, 2H-pyran-2-one, chlorobenzene, 4 cyclopenten-1,3-dione, phenol, benzofuran, 2-chlorohydroquinone and 2-indanone. These concentration profiles have revealed that temperatures of at least 900 degrees C were needed to completely oxidize 2-CP, CO and all other organic byproducts to carbon dioxide. Reaction pathways accounting for the formation of most observed products are proposed. PMID- 16213546 TI - Recovery of phosphates from wastewater using converter slag: Kinetics analysis of a completely mixed phosphorus crystallization process. AB - A phosphorus crystallization process for recovering phosphates was developed using a completely mixed reactor and powdered converter slag as a seed crystal. This completely mixed phosphorus crystallization process achieved a stable and high phosphorus recovery: the average PO4-P removal efficiency during 200 d of operation was 87%, with a range of 70-98%. The apparent volume of the slag doubled due to crystal growth during the long-term phosphorus-removal experiments. The Ca2+ concentration, slag dosage, and temperature were found to govern the phosphorus recovery system for a given condition of pH and hydraulic retention time. The equations for the rate constant and reaction order were obtained by evaluating the model parameters. The model developed in this study was observed to successfully simulate the behavior of effluent PO4-P in a completely mixed phosphorus crystallization reactor over a wide variety of operating conditions of temperature, Ca2+ concentration, and influent PO4-P. Model investigations of design factors suggest that the completely mixed phosphorus crystallization process with influent PO4-P concentrations of less than 10 mg l(-1) could ensure effluent PO4-P concentrations of less than 0.5 and 1.0 mg l(-1) during summer and winter in Korea, respectively. PMID- 16213548 TI - A multispecies study to assess the toxic and genotoxic effect of pharmaceuticals: furosemide and its photoproduct. AB - Pharmaceutical products for humans and animals, as well as their related metabolites end up in the aquatic environment after use. Recent investigations show that concentrations of pharmaceuticals are detectable in the order of ng/l mug/l in municipal wastewater, groundwater and also drinking water. Little is known about the effects, and the hazard of long-term exposure to low concentrations of pharmaceuticals for non-target aquatic organisms. This study was designed to assess the ecotoxicity of furosemide, a potent diuretic agent, and its photoproduct in the aquatic environment. Bioassays were performed on bacteria, algae, rotifers and microcrustaceans to assess acute and chronic toxicity, while the SOS Chromotest and the Ames test were utilized to detect the genotoxic potential of the investigated compounds. A first approach to risk characterization was to calculate the environmental impact of furosemide by measured environmental concentration and predicted no effect concentration ratio (MEC/PNEC). To do so we used occurrence data reported in the literature and our toxicity results. The results showed that acute toxicity was in the order of mg/l for the crustaceans and absent for bacteria and rotifers. Chronic exposure to these compounds caused inhibition of growth population on the consumers, while the algae did not seem to be affected. A mutagenic potential was found for the photoproduct compared to the parental compound suggesting that byproducts ought to be considered in the environmental assessment of drugs. The risk calculated for furosemide suggested its harmlessness on the aquatic compartment. PMID- 16213549 TI - Uptake of polycyclic aromatic hydrocarbons (PAHs) in salt marsh plants Spartina alterniflora grown in contaminated sediments. AB - Polycyclic aromatic hydrocarbon (PAH) concentrations were measured in Spartina alterniflora plants grown in pots of contaminated sediment, plants grown in native sediment at a marsh contaminated with up to 900 microg/g total PAHs, and from plants grown in uncontaminated control sediment. The roots and leaves of the plants were separated, cleaned, and analyzed for PAHs. PAH compounds were detected at up to 43 microg/g dry weight in the root tissue of plants grown in pots of contaminated soil. PAH compounds were detected at up to 0.2 microg/g in the leaves of plants grown in pots of contaminated soil. Concentrations less than 0.004 microg/g were detected in the leaves of plants grown at a reference site. Root concentration factor (RCF) values ranged from 0.009 to 0.97 in the potted plants, and from 0.004 to 0.31 at the contaminated marsh site. Stem concentration factor (SCF) values ranged from 0.00004 to 0.03 in the potted plants and 0.0002 to 0.04 at the contaminated marsh. No correlation was found between the RCF value and PAH compound or chemical properties such as logKOW. SCF values were higher for the lighter PAHs in the potted plants, but not in the plants collected from the contaminated marsh. PAH concentrations in the roots of the potted plants are strongly correlated with soil concentrations, but there is less correlation for the roots grown in natural sediments. Additional plants were grown directly in PAH-contaminated water and analyzed for alkylated PAH homologs. No difference was found in leaf PAH concentrations between plants grown in contaminated water and control plants. PMID- 16213550 TI - Lead, cadmium and arsenic bioavailability in the abandoned mine site of Cabezo Rajao (Murcia, SE Spain). AB - An in vitro method that simulated the physiological conditions of the digestive process was applied to samples taken from an old mining site, providing information on the levels of metals (Cd, Pb and As) that can be ingested and assimilated by humans. Samples were first characterized by determining pH, texture, mineralogical composition and total metal contents. The mean pH value was 5.4, ranging from 3.1 to 8.4. The mean total metal content for Pb was 2,632+/ 59 mg kg(-1), 65+/-1.5 mg kg(-1) for Cd and 279.9+/-9.9 mg kg(-1) for As. Cadmium was the most bioavailable metal both in the stomach and intestinal phases (mean value of 47% and 27.8%, respectively), followed by lead (25.3% and 11.5%) and arsenic (4.9% and 0.5%). A Pearson correlation matrix suggested that pH and mineralogical composition were important factors controlling metal bioavailability from materials in abandoned mining sites. PMID- 16213551 TI - Sorption of small metabolites of nonylphenol polyethoxylates in single and complex systems on aquatic suspended particulate matter. AB - Sorption of nonylphenol (NP), nonylphenol monoethoxylate (NP1EO) and nonylphenol diethoxylate (NP2EO) as well as their binary and ternary mixtures were studied and compared on three simulated suspended particulate matters (SPMs). Sorption dynamics of NP on the three SPMs could be divided into two phases, the rapid sorption phase and the slow sorption phase. A third phase, 'apparent desorption' occurred before the slow sorption phase for NP1EO and NP2EO as well as for all mixtures. Initial sorption rate increased with the OC% content of the SPMs. At low concentration, the sorption of NP, NP1EO and NP2EO (only at low concentration for 3# SPM) followed linear isotherm on the three SPMs. The linear Kd value of NP or NP1EO increased with the OC% content of SPM. In mixtures, sorption of NP, NP1EO and NP2EO increased significantly, and a 'critical point', after which sorption increased significantly, was observed in certain sorption isotherms. PMID- 16213552 TI - Degradation of polychlorinated dibenzo-p-dioxins in aqueous solution by Fe(II)/H2O2/UV system. AB - The photodegradation of polychlorinated dibenzo-p-dioxins (PCDDs), which include tetra- to octa-CDDs (TeCDD, PeCDD, HxCDD, HpCDD and OCDD), was carried out in the presence of Fe(II) and H2O2 mixed reagent. The degradation efficiency was strongly influenced by UV irradiation, and the initial concentrations of H2O2 and Fe(II). An initial TeCDD concentration of 10 ng l(-1) was completely degraded within 20 min under the optimum conditions. All PCDDs tested were successfully degraded by Fe(II)/H2O2/UV treatment and complete degradation of TeCDD, PeCDD and HxCDD was achieved within 120 min. PCDD photodegradation rates decreased with the number of chlorine atoms. The degradation process of TeCDD by this system seems to be initiated by an oxidative reaction (OH* radical attack) because less chlorinated DDs as intermediate products were not detected. From the Frontier electron density calculation, the first OH* radical attack positions on TeCDD were found to be four C atoms neighboring two O atoms. The decomposition of TeCDD gave 4,5-dichlorocatechol as an intermediate product. A TeCDD degradation scheme was proposed based on the identified intermediate and the values of Frontier electron density. Based on these results, Fe(II)/H2O2/UV system could be useful technology for the treatment of wastewater containing persistent pollutants such as dioxins and polychlorinated biphenyls. PMID- 16213553 TI - Identification of hydroxylated metabolites in 2,2',4,4'-tetrabromodiphenyl ether exposed rats. AB - Faeces from day 1-5 of orally administered 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) in rat have been analysed for hydroxylated metabolites. Six hydroxylated tetrabrominated diphenyl ethers, as well as three hydroxylated tribrominated diphenyl ethers found, were structurally identified. They were 2'-hydroxy-2,4,4' tribromodiphenyl ether, 3'-hydroxy-2,4,4'-tribromodiphenyl ether, 4'-hydroxy 2,2',4-tribromodiphenyl ether, 6-hydroxy-2,2',4,4'-tetrabromodiphenyl ether, 2' hydroxy-2,3',4,4'-tetrabromodiphenyl ether, 3-hydroxy-2,2',4,4' tetrabromodiphenyl ether, 5-hydroxy-2,2',4,4'-tetrabromodiphenyl ether, 4' hydroxy-2,2',4,5'-tetrabromodiphenyl ether and 4-hydroxy-2,2',3,4' tetrabromodiphenyl ether. The analysis was performed using gas chromatography mass spectrometry (GC-MS). The identification of the hydroxylated polybrominated diphenyl ether (OH-PBDE) metabolites in the rat faeces was supported by similar relative retention times (RRTs) versus 2,2',3,4,4',5-hexabromodiphenyl ether (BDE 138) on two columns of different polarities compared to the authentic references. The identification of the OH-PBDE metabolites was also supported by full scan electron ionisation mass spectra. Two of the identified OH-PBDE metabolites have identical structures as natural products, which previously have been isolated from marine sponges and an ascidian. PMID- 16213554 TI - Heavy metal contamination from mining sites in South Morocco: 1. Use of a biotest to assess metal toxicity of tailings and soils. AB - Our work was conducted to investigate the heavy metal toxicity of tailings and soils collected from five metal mines located in the south of Morocco. We used the MetPAD biotest Kit which detects the toxicity specifically due to the heavy metals in environmental samples. This biotest initially developed to assess the toxicity of aquatic samples was adapted to the heterogeneous physico-chemical conditions of anthropogenic soils. Contrasted industrial soils were collected from four abandoned mines (A, B, C and E) and one mine (D) still active. The toxicity test was run concurrently with chemical analyses on the aqueous extracts of tailings materials and soils in order to assess the potential availability of heavy metals. Soil pH was variable, ranging from very acidic (pH 2.6) to alkaline values (pH 8.0-8.8). The tailings from polymetallic mines (B and D) contained very high concentrations of Zn (38,000-108,000 mg kg(-1)), Pb (20,412-30,100 mg kg(-1)), Cu (2,019-8,635 mg kg(-1)) and Cd (148-228 mg kg(-1)). Water-extractable metal concentrations (i.e., soil extracts) were much lower but were highly toxic as shown by the MetPAD test, except for soils from mines A, E and site C3 from mine C. The soil extracts from mine D were the most toxic amongst all the soils tested. On this site, the toxicity of soil water extracts was mainly due to high concentrations of Zn (785-1,753 mg l(-1)), Cu (1.8-82 mg l(-1)) and Cd (2.0-2.7 mg l(-1)). The general trend observed was an increase in metal toxicity measured by the biotest with increasing available metal contents in tailings materials and soils. Therefore, the MetPAD test can be used as a rapid and sensitive predictive tool to assess the heavy metal availability in soils highly contaminated by mining activities. PMID- 16213555 TI - Perfluorinated chemicals in selected residents of the American continent. AB - Perfluorinated chemicals (PFCs) are used in multiple consumer products. Perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA), the most widely studied PFCs, may be potential developmental, reproductive, and systemic toxicants. Although PFCs seem to be ubiquitous contaminants found both in humans and animals, geographic differences may exist in human exposure patterns to PFCs. We measured 11 PFCs in 23 pooled serum samples collected in the United States from 1990 through 2002, and in serum samples collected in 2003 from 44 residents from Trujillo, Peru. PFOS and PFOA were detected in all the pooled samples; perfluorohexane sulfonic acid (PFHxS) was detected in 21. Median concentrations were 31.1 micrograms per liter (mug/l, PFOS), 11.6 microg/l (PFOA), and 2 microg/l (PFHxS). The 90th percentile concentrations of PFCs in the 44 Peruvian residents were 0.7 microg/l (PFOS), 0.1 microg/l (PFOA), and <0.3 microg/l (PFHxS). The frequencies of detection were 20% (PFOS), 25% (PFOA), and 9% (PFHxS). The frequent detection of selected PFCs in the pooled samples from the United States and the lack of clear concentration trends based on a year of collection suggest a sustained widespread exposure to these compounds among US residents, at least since the 1990s. By contrast, the much lower frequency of detection and concentration ranges of PFCs in Peru suggest a lower exposure of Peruvians to PFCs compared with North Americans. Genetic variability, diet, lifestyle, or a combination of all these may contribute to the different patterns of human exposure to PFCs in the United States and Peru. PMID- 16213557 TI - Combined effect of microwave and activated carbon on the remediation of polychlorinated biphenyl-contaminated soil. AB - The application of microwave and activated carbon for the treatment of polychlorinated biphenyl (PCB) contaminated soil was explored in this study with a model compound of 2,4,5-trichlorobiphenyl (PCB29). PCB-contaminated soil was treated in a quartz reactor by microwave irradiation at 2450MHz with the addition of granular activated carbon (GAC). In this procedure, GAC acted as microwave absorbent for reaching high temperature and reductant for dechlorination. A sheltered type-K thermocouple was applied to record the temperature rising courses. It was shown that the addition of GAC could effectively promote the temperature rising courses. The determination of PCB residues in soil by gas chromatography (GC) revealed that rates of PCB removal were highly dependent on microwave power, soil moisture content, and the amount of GAC added. GC with mass spectrum (MS) detector and ion chromatography were employed for the analysis of degradation intermediates and chlorine ions, respectively. It was suggested that microwave irradiation with the assistance of activated carbon might be a potential technology for the remediation of PCB-contaminated soil. PMID- 16213556 TI - Heavy metal contamination from mining sites in South Morocco: 2. Assessment of metal accumulation and toxicity in plants. AB - Metalliferous soils cover a relatively large surface area in Morocco, and up to now no hyperaccumulating plants have been identified on these mining or these industrial sites. The aim of this work was to assess the extent of metal accumulation by plants found in three mining areas in southern Morocco with the ultimate goal of finding metal hyperaccumulating species by using the MetPAD biotest. The biotest helps to obtain information on the selective metal toxicity of aqueous extracts from the plants. A strong metal toxicity, as revealed by the biotest is an indication of a hyperaccumulating plant. Toxicity tests were run concurrently with chemicals analyses of metals in plants and their water extracts. The chemical analyses allow the determination of the hyperaccumulated metal(s). Specimens of the plant species mainly growing on and in the vicinity of the three mines were sampled with their corresponding soils. The results show that all plants analyzed had lower heavy metal content and toxicity despite the relatively very high soil concentrations. A comparison of our results with the criterion used to classify the hyperaccumulator plants indicates that plants we collected from mining sites were hypertolerant but not hyperaccumulators. This was confirmed by transfer factors generally lower than 1. Nevertheless, these tolerant plants species can be used as tools for revegetation for erosion control in metals-contaminated sites (phytostabilization). PMID- 16213558 TI - Transfer of nutrients and labile metals from the continent to the sea by a small Mediterranean river. AB - The contribution of small watersheds to coastal pollution in Mediterranean areas is still poorly known, and presents great variations along an hydrological year. This work deals with the characterization and quantification of Al, Fe, Zn, Cu, Pb, Si, NO(3)(-) and organic carbon transported from the continent to the sea by a small coastal river in the French Mediterranean area. Transported species were characterized both during rainy and dry periods. During non-rainy periods (base flow), waters showed a low content of contaminants, whereas during heavy rain events, following dry periods, some metals and organic carbon reached concentrations that could affect biological populations. These contaminants were mainly found in the particulate fraction, originating from the runoff of surface waters, which represents the main process of pollution in urban areas. PMID- 16213559 TI - Biostimulation and bioaugmentation enhances aerobic biodegradation of dichloroethenes. AB - The accumulation of dichloroethenes (DCEs) as dominant products of microbial reductive dechlorination activity in soil and water represent a significant obstacle to the application of bioremediation as a remedial option for chloroethenes in many contaminated systems. In this study, the effects of biostimulation and/or bioaugmentation on the biodegradation of cis- and trans-DCE in soil and water samples collected from contaminated sites in South Africa were evaluated in order to determine the possible bioremediation option for these compounds in the contaminated sites. Results from this study indicate that cis- and trans-DCE were readily degraded to varying degrees by natural microbial populations in all the soil and water samples tested, with up to 44% of cis-DCE and 41% of trans-DCE degraded in the untreated soil and water samples in two weeks. The degradation rate constants ranged significantly (P<0.05) between 0.0938 and 0.560 wk(-1) and 0.182 and 0.401 wk(-1), for cis- and trans-DCE, respectively, for the various treatments employed. A combination of biostimulation and bioaugmentation significantly increased the biodegradation of both compounds within two weeks; 14% for cis-DCE and 18% for trans-DCE degradation, above those observed in untreated soil and water samples. These findings support the use of a combination of biostimulation and bioaugmentation for the efficient biodegradation of these compounds in contaminated soil and water. In addition, the results clearly demonstrate that while naturally occurring microorganisms are capable of aerobic biodegradation of cis- and trans DCE, biotransformation may be affected by several factors, including isomer structure, soil type, and the amount of nutrients available in the water and soil. PMID- 16213560 TI - Uptake and distribution of Zn, Cu, Cd, and Pb in an aquatic plant Potamogeton natans. AB - A better understanding of metal uptake and translocation by aquatic plants can be used to enhance the performance of constructed wetland systems for stormwater treatment. Specifically, this study examines whether the uptake of Zn, Cu, Cd, and Pb by Potamogeton natans is via the leaves, stems, or roots, and whether there is translocation from organs of uptake to other plant parts. Competition between the metals at uptake and at the level of the cell wall-bound part of the metals accumulated in stem and leaf tissue was also examined. The results show that Zn, Cu, Cd, and Pb were taken up by the leaves, stems, and roots, with the highest accumulation found in the roots. At the elevated metal concentrations common in stormwater the uptake of Cu, but not of Zn, Cd, or Pb, by the roots was somewhat limited at uptake due to competition with other metals. Between 24% and 59% of the metal content was bound to the cell walls of the plant. Except in the case of Pb, the cell wall-bound fraction was generally smaller in stems than in leaves. No translocation of the metals to other parts of the plant was found, except for Cd which was translocated from leaf to stem and vice versa. Dispersion of metals from sediment to water through P. natans is therefore unlikely. PMID- 16213561 TI - Assessing the organic composition of urban surface films using nuclear magnetic resonance spectroscopy. AB - Recently it has been shown that urban surfaces are covered with a thin film which mediates the fate, distribution and accumulation of semi-volatile organic compounds in the environment. In this study we apply a combination of solution, semi-solids, and solid state nuclear magnetic resonance (NMR) methods to provide a general overview of the organic constituents. In surface film collected from 30 m2 of outside windows over an area of 12 km2 in downtown Toronto, we roughly estimate that the organic carbon is approximately 35% carbohydrate, approximately 35% aliphatics, approximately 20% aromatics, and approximately 10% carbonyl groups. Various aliphatic groups can be identified including a number of acids, alcohols, alkanes, and alkenes. Also, numerous intact aliphatic esters are apparent that have not been observed before, as well as carbohydrates. The aromatic species include a small portion that appears to be derived from a polymer of styrene, in addition a larger fraction is consistent with polyhydroxylated PAH derived material, although this assignment is tentative and based solely on 1-D NMR data only. In addition, signals from polybutadiene are present and while accurate quantification is not possible, it appears that this polymer may be up to a few percents by weight of the total organic material. PMID- 16213562 TI - Montmorillonite-Cu(II)/Fe(III) oxides magnetic material as adsorbent for removal of humic acid and its thermal regeneration. AB - In this work, the adsorption features of montmorillonite and the magnetic properties of Cu(II)/Fe(III) oxides were combined in a material to produce magnetic adsorbent, which can be separated from the medium by a simple magnetic process after adsorption. The magnetic material is effective for the removal of humic acid. At pH 6.1, 96% removal was observed from 4.4 mg l(-1) humic acid solution containing 0.02 M NaCl. The adsorption is pH and ionic strength dependent. Adsorption is favored at lower pH values and dissolved NaCl can enhance the adsorption. The adsorption mechanism of humic acid to the magnetic material was suggested to be the ligand exchange reaction between carboxylic groups of humic acid molecules and the magnetic material surface. The magnetic material can be thermally regenerated. The temperature and time required to achieve good regeneration efficiency were determined to be 300 degrees C and 3 h, respectively. The regenerated adsorbent is still magnetic and approximately has as high specific saturation magnetization and good adsorption capacities as the as-prepared adsorbent. PMID- 16213563 TI - Degradation of toxaphene in aged and freshly contaminated soil. AB - Degradation of toxaphene in soil from both newly contaminated (from Sweden) and aged spills (from Nicaragua) were studied. The newly contaminated soil contained approximately 11 mg kg(-1) toxaphene while the aged Nicaraguan soil contained approximately 100 mg kg(-1). Degradation was studied in anaerobic bioreactors, some of which were supplied with lactic acid and others with Triton X-114. In this study we found that the lower isomers Parlar 11, 12 were degraded while the concentration of isomer Parlar 15 increased. This supported an earlier evaluation which indicated that less chlorinated isomers are formed from more heavily isomers. Lactic acid when added to the soil, interfere with the degradation of toxaphene. Lactic acid was added; several isomers appeared to degrade rather slowly in newly contaminated Swedish soil. The Swedish soil, without any external carbon source, showed the slowest degradation rate of all the compounds studied. When Triton X-114 at 0.4 mM was added, the degradation rate of the compounds increased. This study illustrates that biodegradation of toxaphene is a complex process and several parameters have to be taken into consideration. Degradation of persistent pollutants in the environment using biotechnology is dependent on bioavailability, carbon sources and formation of metabolites. PMID- 16213564 TI - The exploration of potassium ferrate(VI) as a disinfectant/coagulant in water and wastewater treatment. AB - This paper aims to explore potassium ferrate(VI) (K2FeO4) as an alternative water treatment chemical for both drinking water and wastewater treatment. The performance of potassium ferrate(VI) was evaluated in comparison with that of sodium hypochlorite (NaOCl) and that of NaOCl plus ferric sulphate (FS) or alum (AS). The dosages of ferrate(VI), NaOCl and FS/AS and sample pH values were varied in order to investigate the effects of these factors on the treatment performance. The study demonstrates that in drinking water treatment, ferrate(VI) can remove 10-20% more UV(254)-abs and DOC than FS for the same dose compared for natural pH range (6 and 8). The THMFP was reduced to less than 100 microg l(-1) by ferrate(VI) at a low dose. In addition to this, ferrate(VI) can achieve the disinfection targets (>6 log10 inactivation of Escherichia coliform (E. coli)) at a very low dose (6 mg l(-1) as Fe) and over wide working pH in comparison with chlorination (10 mg l(-1) as Cl2) plus coagulation (FS, 4 mg l(-1) as Fe). In wastewater treatment, ferrate(VI) can reduce 30% more COD, and kill 3log10 more bacteria compared to AS and FS at a similar or even smaller dose. Also, potassium ferrate(VI) can produce less sludge volume and remove more pollutants, which could make sludge treatment easier. PMID- 16213565 TI - Isomeric analysis of PCDD/PCDF in waste incinerator fly ash by GC-MS/MS. AB - The development of an analytical method for the analysis of PCDD/PCDF in fly ash using a bench analytical system comprising of a gas chromatograph fitted with an ion trap detector operated in the tandem MS mode is described. The optimum settings for the most important MS/MS parameters are given, including those for the less reported mono- to tri-chlorinated dioxin and furan congener groups. Fly ash samples from three waste incineration plants representing a de-commissioned 1970s plant design, an upgraded and still operating plant originally designed in the 1970s, and a modern 1990s design operating plant have been analysed for PCDD/PCDF. The fly ash samples were analysed for PCDD/PCDF using the methods developed and the total PCDD/PCDF, I-TEQ values and isomeric profiles are reported. The fly ash from the older de-commissioned incinerator had very significantly higher concentrations of PCDD/PCDF compared to the modern incinerator fly ash. PMID- 16213566 TI - Enantiomeric fractions and congener specific determination of polychlorinated biphenyls in eggs of predatory birds from Donana National Park (Spain). AB - The content of 30 polychlorinated biphenyls (PCBs) and the enantiomeric fractions of 10 chiral PCBs were determined in 17 infertile eggs from three different predatory bird species collected in Donana National Park (DNP, Spain) in the period 1999-2000. The highest PCB concentration was found in eggs from red kites (0.52-110 microg/g on a fresh weight basis, f.w.) followed by buzzard (0.08-13 microg/g f.w.) and booted eagle (0.10-1.5 microg/g f.w.). Seventy-five percent of the red kite eggs had PCB levels above 4.7 microg/g f.w., which is associated in the literature with reproductive failure. This could be related to the fact that red kite populations have decreased by more than 50% in the last five years in DNP. PCBs # 138, 153, and 180 were the most abundant in all cases. This is the first time that atropisomers of 10 chiral PCBs (PCBs # 45, 84, 91, 95, 132, 135, 136, 149, 174, and 176) in predatory bird eggs have been performed. The enantiomeric fractions (EFs) for most PCBs investigated were non-racemic (EF not = 0.5), ranging from 0.05 to 0.95. The results suggested that predatory birds, mainly red kite species, are highly polluted by PCBs, and PCBs # 95, 132, 135, 136, and 174 strongly deviate from the racemic-mixture values. PMID- 16213567 TI - Modification of UNIFAC parameter table Revision 5 for representation of aqueous solubility and 1-octanol/water partition coefficient for POPs. AB - In order to represent aqueous solubility and 1-octanol/water partition coefficient for POPs (persistent organic pollutants) by the UNIFAC model, two pairs of group interaction parameters in Revision 5 of the UNIFAC parameter table were modified. First, the pair of interaction parameters between the aromatic carbon-chloride (ACCl) and water (H2O) groups were corrected by minimizing the deviation between the experimental and calculated values of these properties for chlorinated benzenes, polychlorinated biphenyls, and polychlorinated dibenzo-p dioxins and dibenzofurans. The corrected interaction parameters provided a better representation of both properties than the calculation results obtained in early works using the UNIFAC model. Second, the unknown pair of interaction parameters between the chloroalkene (Cl(C=C)) and H2O groups, which are required for calculating those properties of the other five POPs (aldrin, chlordane, endrin, dieldrin, and heptachlor), were newly determined from the experimental data on their properties. Finally, this study shows that Revision 5 could also predict solubility of POPs in some organic solvents. The modified parameter table is first suggested as the UNIFAC parameter table applicable to various phase equilibria including aqueous or nonaqueous solubility and partition coefficient of POPs. PMID- 16213568 TI - Model comparison for risk assessment: a case study of contaminated groundwater. AB - Many environmental multimedia risk assessment models have been developed and widely used along with increasing sophistication of the risk assessment method. Despite of the considerable improvement, uncertainty remains a primary threat to the credibility of and users' confidence in the model-based risk assessments. In particular, it has been indicated that scenario and model uncertainty may affect significantly the assessment outcome. Furthermore, the uncertainty resulting from choosing different models has been shown more important than that caused by parameter uncertainty. Based on the relationship between exposure pathways and estimated risk results, this study develops a screening procedure to compare the relative suitability between potential multimedia models, which would facilitate the reduction of uncertainty due to model selection. MEPAS, MMSOILS, and CalTOX models, combined with Monte Carlo simulation, are applied to a realistic groundwater-contaminated site to demonstrate the process. It is also shown that the identification of important parameters and exposure pathways, and implicitly, the subsequent design of uncertainty reduction and risk management measures, would be better-formed. PMID- 16213569 TI - Assessment of the risk posed by the antifouling booster biocides Irgarol 1051 and diuron to freshwater macrophytes. AB - Antifouling paints are used to reduce the attachment of living organisms to the submerged surfaces of ships, boats and aquatic structures, usually by the release of a biocide. Two 'booster' biocides in common use are the triazine herbicide Irgarol 1051 (N-2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine), and diuron (1-(3,4-dichlorophenyl)-3,3-dimethylurea), which are designed to inhibit algal photosynthesis. Previous research has been directed at the effects of these compounds in marine and estuarine environments. In 2001 we sampled the main rivers and shallow freshwater lakes (Broads) of East Anglia UK for Irgarol 1051, its metabolite GS26575 (2-methylamino-4-tert-butylamino-6-amino-s-triazine) and diuron in order to establish the baseline environmental concentrations of these compounds in freshwater systems of eastern UK and to investigate their possible effects on aquatic plants. Irgarol 1051, GS26575 and diuron were found in water samples collected from 21 locations. The highest concentrations were found in the Norfolk and Suffolk Broads in May. The rivers Great Ouse, Wissey, Bure and Yare also contained all three compounds, as did the Great Ouse Cut-off Channel. The toxicity of these biocides to three macrophyte species (Apium nodiflorum, Chara vulgaris, and Myriophyllum spicatum) was investigated. Deleterious effects on relative growth rate, the maximum quantum efficiency (Fv/Fm) of photosystem II and, for Apium, root mass production were found. C. vulgaris was generally most sensitive; growth, especially of roots, was strongly affected in A. nodiflorum; growth rate of M. spicatum was sensitive to diuron. No observed effect concentrations (NOEC) were interpolated using standard toxicological analysis. These were compared with measured environmental concentrations (MEC) to determine the ranges of risk quotients (MEC/NOEC). Both Irgarol 1051 and diuron represented significant risks to A. nodiflorum and C. vulgaris in this area. PMID- 16213570 TI - Extractability and bioavailability of zinc over time in three tropical soils incubated with biosolids. AB - Phytotoxicity of heavy metal is the primary concern in applying biosolids (sewage sludge) to agricultural land. This study evaluates the changes in chemical speciation of Zn in three tropical soils of Taiwan measured with sequential extraction over a one-year period. Biosolids were applied to the soils at application rates of 10, 50 and 100 Mg ha(-1), and correlated diethylene triamine pentaacetic acid (DTPA) and sequential extraction as extract for prediction of Zn bioavailability to Chinese cabbage (Brassica chinensis L.). Experimental results indicated that the exchangeable (F1) and Fe-Mn oxide (F3) fractions in the sequential extractions increased with application rate of biosolids in the soils over time. Large amounts of Zn in the soils following the cessation of biosolids application were identified as soluble and were adsorbed by Fe-Mn oxides. The organically bound Zn, which is associated with readily decomposable carbon, is in limited amounts in the biosolid-treated soils. The DTPA-extractable concentrations of Zn in all biosolid-treated soils decreased over the time. A positive and significant correlation (r(2) = 0.96) was found between the Zn concentrations extracted with DTPA and sum of F1 and carbonate-bound (F2) fractions in the sequential extractions. Additionally, the concentrations of Zn extracted with DTPA were strongly correlated with the concentrations of Zn in the shoots of Chinese cabbages, indicating that F1+F2 in the sequential extractions was reliable for predicting Zn bioavailability to Chinese cabbage in the biosolid treated soils. PMID- 16213571 TI - Toxicity of emerging energetic soil contaminant CL-20 to potworm Enchytraeus crypticus in freshly amended or weathered and aged treatments. AB - We investigated the toxicity of an emerging polynitramine energetic material hexanitrohexaazaisowurtzitane (CL-20) to the soil invertebrate species Enchytraeus crypticus by adapting then using the Enchytraeid Reproduction Test (ISO/16387:2003). Studies were designed to develop ecotoxicological benchmark values for ecological risk assessment of the potential impacts of accidental release of this compound into the environment. Tests were conducted in Sassafras Sandy Loam soil, which supports relatively high bioavailability of CL-20. Weathering and aging procedures for CL-20 amended into test soil were incorporated into the study design to produce toxicity data that better reflect soil exposure conditions in the field compared with the toxicity in freshly amended soils. Concentration-response relationships for measurement endpoints were determined using nonlinear regressions. Definitive tests showed that toxicities for E. crypticus adult survival and juvenile production were significantly increased in weathered and aged soil treatments compared with toxicity in freshly amended soil, based on 95% confidence intervals. The median effect concentration (EC50) and EC20 values for juvenile production were 0.3 and 0.1 mg kg-1, respectively, for CL-20 freshly amended into soil, and 0.1 and 0.035 mg kg-1, respectively, for weathered and aged CL-20 soil treatments. These findings of increased toxicity to E. crypticus in weathered and aged CL-20 soil treatments compared with exposures in freshly amended soils show that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information on ecotoxicological effects of emerging energetic contaminants in soil. PMID- 16213572 TI - Formation of dioxins from combustion of polyvinylidene chloride in a well controlled incinerator. AB - Polyvinylidene chloride (PVDC; polymer of 1,1-dichloroethylene) was combusted with paper in a well-controlled, small-scale incinerator at an average grate temperature of 700 degrees C, and then dioxins (PCDDs, PCDFs, and coplanar-PCBs) formed in the exhaust gases were analyzed by gas chromatography/mass spectrometry. PVDC lowered the combustion temperature due to its less flammable character. The amount of total dioxins (PCDDs+PCDFs+coplanar-PCBs) formed in the exhaust gas was 58.0 ng/g of a combustion sample and its toxicity equivalency quantity (TEQ) value was 0.64 ng-TEQ/g. The amount of PCDDs formed in the sample ranged from 2.33 ng/g (Cl(8)-isomer) to 0.048 ng/g (Cl(1)-isomer). The lower the number of chloride, the less production of PCDDs. On the other hand, there was no relation between the number of chloride and PCDF formation. The amount of PCDFs formed in the sample ranged from 8.02 ng/g (Cl(2)-isomer) to 4.46 ng/g (Cl(8) isomer). A polyvinylchloride (PVC) sample produced 207 ng/g of total dioxins and a PVDC sample produced 57.4 ng/g of total dioxins when they were combusted under the same conditions. An approximately equal composition of dioxin isomers was formed from PVDC and PVC samples. Paper was found to contribute to PCDF formation when it was combusted with plastics. PMID- 16213574 TI - Continuous growth of telomerase-immortalised fibroblasts: how long do cells remain normal? AB - Previously, we reported successful immortalisation following hTERT introduction in primary human fibroblasts, strain VH25. Since one subclone in that study developed some abnormalities, we decided to study eight additional independent immortalised clones to get an indication of the frequency and type of abnormalities that develop after hTERT-mediated immortalisation. We show that although some cell lines can maintain a normal phenotype for 500 population doublings (PDs), in four clones after 150-300PDs changes developed in basal and radiation-induced p53 and p21(WAF-1,CIP-1) levels. Our experiments demonstrate that, after prolonged culture, cells with abnormalities in cell cycle control parameters can take over the population. This calls for caution when working with hTERT-immortalised cells in vitro as well as in vivo. PMID- 16213575 TI - Identification of differentially expressed proteins in senescent human embryonic fibroblasts. AB - Normal human fibroblasts undergo a limited number of divisions in culture, a process known as replicative senescence (RS). Although several senescence specific genes have been identified, analysis at the level of protein expression can provide additional insights into the mechanisms that regulate RS. We have performed a proteomic comparison between young and replicative senescent human embryonic WI-38 fibroblasts and we have identified 13 proteins, which are differentially expressed in senescent cells. Some of the identified proteins are components of the cellular cytoskeleton, while others are implicated in key cellular functions including metabolism and energy production, Ca(2+) signalling, nucleo-cytoplasmic trafficking and telomerase activity regulation. In summary, our analysis contributes to the list of senescence-associated proteins by identifying new biomarkers and provides novel information on functional protein networks that are perturbed during replicative senescence of human fibroblast cultures. PMID- 16213576 TI - Investigation of extraction and clean-up procedures used in the quantification and stable isotopic characterisation of PAHs in contaminated urban soils. AB - Four different extraction methods, soxhlet, soxtherm, sonication and accelerated solvent extraction (ASE), were used to isolate the 16 priority pollutant PAHs from a certified reference soil (LGC 6140) and from a contaminated soil (BG CLR 17). Based on SIM-GC-MS results, all methods were found to give accurate and highly reproducible concentration data. There was, however, significant between method and sometimes within-method variability in the stable carbon isotope signatures obtained for individual PAHs from the contaminated soil (BG CLR 17) using GC-C-IRMS. When two clean-up procedures, silica/dichloromethane and alumina/hexane/toluene, were used to remove co-extracted material, however, it was found that ASE gave the more consistent and reproducible stable carbon isotope data. These findings are likely to be of importance for the characterisation of natural and anthropogenic organic matter and, in particular, in source identification and apportionment studies. PMID- 16213577 TI - Exogenous enzyme supplements to promote treatment efficiency in constructed wetlands. AB - Extracellular enzymes play a central role in the breakdown of organic pollutants. In wetlands constructed to treat wastewaters, supplementing the naturally occurring soil enzymes may result in faster pollutant removal, or breakdown of novel pollutants, but only if the added enzymes could retain their catalytic activity. In this study, the persistence of exogenous enzyme supplements was investigated. Adding cellobiohydrolase and beta-glucosidase to sterilised soil increased enzyme activity (range 375-4210%); although the increased activity began to decline after just 10-15 days. Thus, without an active microbial population, enhanced enzyme activity is unlikely to be long lived. However, with the naturally occurring soil microbes present to maintain the improved biodegradative capacity, cellobiohydrolase and beta-glucosidase additions created significant increases (range 173-530%) in activity and these persisted for more than 6 weeks. These findings therefore support the proposal that enzyme additions can enhance enzymic biodegradation processes, and suggest that this may be achieved primarily through a 'pump-priming' mechanism. PMID- 16213579 TI - The effect of filter vent blocking and smoking topography on carbon monoxide levels in smokers. AB - Two studies were conducted to examine the effect of filter vent blocking and smoking topography on carbon monoxide (CO) levels in smokers. In Study 1, 12 participants smoked two types of cigarettes (Marlboro Light and Carlton 100) under two types of blocking conditions (unblocked and half-blocked) while using a smoking topography device. Participants were restricted to 8 puffs, separated by 45 s. Significant main effects of CO boost for cigarette type and blocking condition replicated previous findings. A significant increase in CO boost for the Marlboro Light blocked condition is a novel finding for this best-selling brand. That result and the finding that topography measures did not predict CO boost made us question the reliability of CO boost. In Study 2, we examined the reliability of CO boost by recruiting 12 participants to smoke three unblocked Carlton 100 cigarettes in one session and three half-blocked in another. CO boost was significantly greater for the blocked sessions compared to the unblocked and CO boost did not differ within session, thus supporting the reliability of the measure. When participants do not switch brands within a session, smoking topography measures are predictive of CO boost. PMID- 16213578 TI - The effects of restraint stress on nociceptive responses induced by formalin injected in rat's TMJ. AB - It has been reported that stress can alter nociception from superficial tissues, such as skin and subcutaneous region. However, the influence of stress on an experimental deep nociception model is not understood. In this study, the temporomandibular joint (TMJ) formalin test was used to evaluate the effects of acute and chronic restraint stress on nociceptive responses in rats. Animals were initially submitted to one session of acute restraint stress (1 h) or exposed to chronic stress (40 days-1 h/day). Then, animals were killed immediately to collect blood for hormonal determinations by radioimmunoassay, or submitted to the TMJ formalin test to evaluate nociception. Rats submitted to acute restraint presented a performance similar to unstressed controls in the TMJ formalin test, whereas chronically stressed rats showed an increase in nociceptive responses. After 40 days of restraint, morphine was injected i.p. (1, 5 mg/kg or saline). The stressed rats displayed decreased morphine effects on nociception compared to unstressed controls. These findings suggest that repeated stress can produce hyperalgesia, which is, at least in part, due to alterations in the activity of opioid systems. This model may help elucidate the underlying neural mechanisms that mediate the effects of repeated stress on orofacial pain. PMID- 16213580 TI - Parental influences on young people's sexual behaviour: a longitudinal analysis. AB - Both family structure and processes have been associated with young people's sexual behaviour, but most studies are cross-sectional and focus on only one outcome: age at first intercourse. This paper uses longitudinal data from a survey of Scottish teenagers (N=5041) to show how low parental monitoring predicts early sexual activity for both sexes (with some reverse causation), and for females it also predicts more sexual partners and less condom use. A lot of spending money also predicts early sexual activity and, for males, having more sexual partners. Comfort talking with parents about sex, however, seems to bear little relationship to sexual behaviour. PMID- 16213581 TI - A 5-7 year in vivo study of high-strength hydroxyapatite/poly(L-lactide) composite rods for the internal fixation of bone fractures. AB - The biocompatibility and biodegradation of hydroxyapatite (HA)/poly(L-lactide) (PLLA) composite bone implant rods were studied using two types of HA particles as reinforcing fillers, uncalcined-HA (u-HA) or calcined-HA (c-HA). Composite rods of u-HA/PLLA and c-HA/PLLA were implanted into the distal femurs of 25 rabbits. Four of these rabbits lived for more than 5 years after implantation, with one living for 7 years 4 months. Samples were retrieved after the rabbits died naturally and specimens were examined by light microscopy and scanning electron microscopy (SEM). For u-HA/PLLA, the shrinkage of the rods progressed further after 4 years, and some were almost completely resorbed. We could not find traces of the rod in some sections from more than 6 years. Remodeled bone encircled the residual material, and trabecular bone bonded directly to the rod was found toward the center. For u-HA/PLLA 30%, results were similar to u-HA/PLLA 40%, and the cross-sectional area had decreased by 4.0% at over 7 years. Most of the PLLA had been released from the rods. Neither obvious osteolytic nor osteoarthritic changes were found after 4 years. PMID- 16213582 TI - The Hsp90 inhibitor 17-allylamide-17-demethoxygeldanamycin induces apoptosis and differentiation of Kasumi-1 harboring the Asn822Lys KIT mutation and down regulates KIT protein level. AB - Heat shock protein 90 (Hsp90) serves as a chaperone for a number of cell signaling proteins, including many tyrosine and serine/threonine kinases, which are involved in proliferation and/or survival. The benzoquinone ansamycin geldanamycin has been shown to bind to Hsp90 and to specifically inhibit this chaperone's function, resulting in client protein destabilization. 17-Allylamino 17-demethoxygeldanamycin (17-AAG) is a chemical derivative of geldanamycin. KIT is the receptor for stem cell factor (SCF) and required for normal hematopoiesis. Mutations in c-Kit result in ligand-independent tyrosine kinase activity and uncontrolled cell proliferation. Kasumi-1 is t(8;21) acute myeloid leukemia (AML) cell line harboring mutated KIT with Asn822Lys substitution. Our present studies demonstrate that 17-AAG inhibits Kasumi-1 cells proliferation and exerts apoptosis- and differentiation-inducing effects in a dose- and time-dependent manner. The growth-inhibitory IC50 value for 17-AAG treatment is 0.62mumol/L. Characteristic apoptotic features were confirmed by morphology, internucleosomal DNA fragmentation, and annexin V staining. 17-AAG also causes the G0/G1 block of Kasumi-1 cells. Significantly, 17-AAG-induced apoptosis of Kasumi-1 cells is associated with a decline in KIT protein level. Our findings strongly suggest that 17-AAG might be an effective therapeutic agent targeting AML cells harboring mutated KIT. PMID- 16213583 TI - Sorcin, an important gene associated with multidrug-resistance in human leukemia cells. AB - Sorcin, or soluble resistance-related calcium-binding protein, is a 22kD calcium binding protein initially identified in many mutli-drug resistant (MDR) cell lines. We previously observed by gene profiling that sorcin is significantly up regulated in a doxorubicin-induced MDR leukemia cell line, K562/A02, over its parent cells. We have also demonstrated that the level of sorcin expression in leukemia patients correlates not only directly with that of the mdr1 gene, but also inversely with patients' response to chemotherapies and overall prognosis. In this report, we have carried out experiments to dissect out the contribution of sorcin by itself to drug resistant phenotype in K562 cells. Overexpression of sorcin protein by gene transfection in K562 cells resulted in increased drug resistance, from 4.1- to 22.5-fold, to a variety of chemotherapeutic agents, including doxorubicin, etoposide, homoharringtonine and vincristine. On the other hand, inhibition of sorcin expression in both MDR K562/A02 and the sorcin transfected K562 cells with sorcin-targeting small interfering RNA led to varying extent of reversal of drug resistance. These results confirm that sorcin is an important gene associated with the development of MDR in leukemia cells. PMID- 16213584 TI - HA14-1, a small molecule Bcl-2 antagonist, induces apoptosis and modulates action of selected anticancer drugs in follicular lymphoma B cells. AB - The BCL-2 overexpression is a hallmark of follicular lymphoma (FL). Since patients with FL often suffer from resistant to chemotherapy refractory disease, the development of new regimens is required. Herein, we analyze for the first time the effects of a B-cell lymphoma 2 (Bcl-2) antagonist, HA14-1, alone and in combination with antineoplastic agents commonly used against follicular lymphoma, in human FL cell lines with t(14;18). All cell lines tested were sensitive to HA14-1-induced cytotoxicity and apoptosis, as depicted by morphological changes, SYTO16/PI staining, oligonucleosomal DNA fragmentation and loss of Deltapsi(m). Moreover, HA14-1 significantly enhanced dexamethasone- and doxorubicin-mediated (in schedule independent and dependent manner, respectively), but not vincristine mediated cytotoxicity and apoptosis. PMID- 16213585 TI - Assessment of Godavari estuarine mangrove ecosystem through trace metal studies. AB - Trace metals (Fe, Mn, Cu, Co, Ni. Pb, Cd and Cr) analyses in particulate matter and sediments were carried out at 16 different stations in the Godavari mangrove ecosystem covering the three regions viz., Kakinada (KKD) bay, Goutami-Godavari estuary (GGE) and Coringa and Gaderu mangrove environments (CGME). Higher concentrations of trace metals in particulate matter and in sediments were noticed in the mangrove environment compared to the KKD bay and GGE region. The significant correlations observed between several couples of metals in different regions as well as between numbers of heavy metals (Fe, Mn, Zn) with Total Suspended Matter (TSM) in particulate organic matter could be an indication of the role of the complexation with organic matter in the distribution pattern of these metals. Further significant correlations between organic carbon (OC) and Cr, Co, Pb, Cu and Mn in sediments of all the three regions indicate that organic matter acts as a metal carrier. The values of Pollution Load Index (PLI) were low and varied between 0.34 and 0.68 in KKD bay, 0.47 and 0.49 in GGE and, from 0.62 to 1.03 in CGME area, which indicate that the estuarine ecosystem is not of pollution concern under the present scenario. PMID- 16213586 TI - Studies on heavy metal accumulation in aquatic macrophytes from Sevan (Armenia) and Carambolim (India) lake systems. AB - Aquatic macrophytes are unchangeable biological filters and they carry out purification of the water bodies by accumulating dissolved metals and toxins in their tissue. In view of their potential to entrap several toxic heavy metals, 45 macrophytes belonging to 8 families collected from two different physiographic locations (36 from Sevan Lake, Armenia; 9 from Carambolim Lake, Old Goa, India) were studied for estimation of 14 heavy metals. The study was aimed at understanding the importance of these macrophytes in accumulation of toxic metals and controlling the heavy metal pollution and suggesting the remedial measures, if any, for the preservation and restoration of lake ecosystem. Inductively Coupled Plasma-Atomic Emission Spectrometric (ICP-AES) analyses of these aquatic macrophytes have shown the importance of aquatic macrophytes in accumulation of heavy metals and maintaining the clarity of water bodies beside their role in trophic systems. Accumulation of most of the heavy metals was higher in root system. The representative macrophytes from two different physiographic locations show similar trends and order in accumulating different metals generally. Of the 14 metals investigated, 9 (Ca, Fe, Al, Cr, Cu, Ba, Ti, Co and Pb) showed higher rates of accumulation in the root whereas 3 (Mn, Zn and Mg) showed more accumulation in stem and 1 (Ca) showed higher accumulation in the leaves. In most of the samples Cu was accumulated more in the roots (50+/-47.15 microg/g) and less in flowers (9.52+/-3.97 microg/g). Occurrence of heavy metal was much higher in macrophytes of Sevan Lake than that of the Carambolim Lake. The accumulation of 14 elements was in order of Ca>Mg>Fe>Al>Mn>Ba>Zn>Ti>Cu>Cr>Co>Ni>Pb>Cd. The present study revealed that the aquatic macrophytes play a very significant role in removing the different metals from the ambient environments. They probably play a major role in reducing the effect of high concentration of heavy metals. Therefore, the macrophyte community of the Sevan Lake area needs to be protected and restored on a priority basis. Accumulation of highly toxic metals like--Cr, Cd, Pb and Ni was lower as compared to the essential metals like Ca, Fe and Mn in all the macrophytes from both the lake systems, consequently high metal concentrations observed in both the areas may not directly reflect on the pollution level. PMID- 16213587 TI - Glutamic acid reverses Pb2+-induced reductions of NMDA receptor subunits in vitro. AB - The objective of this study is to determine the effects of Pb2+ on N-methyl-d aspartate receptor (NMDAR) subunits--NR1C1, NR2A and NR2B in primary cultured neuronal cells. We hypothesize that L-glutamic acid (GA) reverses Pb2+-induced NMDAR damage. Neuronal cells were isolated from the fetus brain at 18-20th day of gestation of pregnant Sprague Dawley (SD) rats. All experiments were included three independent cell preparations (N=3). The neuronal cells were exposed to Pb2+ (10(-10), 10(-9), 10(-8) and 10(-7)M) for 24 h. Neurons were pretreated with NMDAR agonist--L-glutamic acid (GA) (200 microM) and antagonists dizocipine (MK 801, 50 nM) for 1h and then exposed to 10(-7)M of Pb2+ for 24 h. Finally, GA at 2, 0.2 and 0.02 mM was incubated with neurons prior to Pb2+ exposure. Aliquots of NR1, NR2A and NR2B proteins from cell homogenate were immunoprecipitated with protein A agarose and detected by Western blotting. The addition of GA unconventionally reversed the reductions of NMDAR by Pb at protein levels, whereas MK-801 exacerbated Pb2+-induced damage. The protection by GA against Pb2+ induced reduction of NMDAR was dose-dependent. These findings suggest that the administration of GA may be a potential approach to intervene the Pb2+-induced NMDAR alterations. PMID- 16213589 TI - Oxidative DNA cleavage mediated by a new copper (II) terpyridine complex: crystal structure and DNA binding studies. AB - The copper (II) complex [Cu(Itpy)(2)](ClO(4))(2) (1), (Itpy=imidazole terpyridine) has been synthesized and structurally characterized. Crystal structure of the complex shows the complex to be a monomeric copper (II) species with two Itpy ligands coordinated to the metal ion to give a six coordinate complex. The complex has a distorted octahedral geometry with axial elongation. Variable temperature crystal structure data shows dynamic nature of the Jahn Teller distortion. The complex is an avid DNA binder with a binding constant of 4.26+/-0.20x10(3)M(-1). Observed changes in the viscosity and circular dichroic spectrum of calf thymus DNA solution in the presence of complex 1 suggests intercalative binding of complex 1 to DNA. The complex cleaves supercoiled pBR322 DNA oxidatively in the presence of hydrogen peroxide. PMID- 16213588 TI - Voltammetry as a virtual potentiometric sensor in modelling of a metal-ligand system and refinement of stability constants. Part 4. An electrochemical study of NiII complexes with methylene diphosphonic acid. AB - The Ni(II)-MDP-OH system (MDP=methylene diphosphonic acid) and stability constants of complexes formed at ionic strength 0.15M at 298K were established by direct current polarography (DCP) and glass electrode potentiometry (GEP). The final M-L-OH model could only be arrived to by employing recent concept of virtual potentiometry (VP). VP-data were generated from non-equilibrium and dynamic DC polarographic technique. The VP and GEP data were refined simultaneously by software dedicated to potentiometric studies of metal complexes. Species distribution diagrams that were generated for different experimental conditions employed in this work assisted in making the final choice regarding the metal-ligand model. The model established contains ML, ML(2), ML(OH) and ML(OH)(2) with stability constants, as logbeta, 7.94+/-0.02, 13.75+/ 0.02, 12.04 (fixed value), and 16.75+/-0.05, respectively. It has been demonstrated that virtual potential must be used in modelling operations (predictions of species formed) when a polarographic signal decreases significantly due to the formation of polarographically inactive species (or formation of inert complexes). The linear free energy relationships that included stability constant logK(1) for Ni(II)-MDP established in this work together with other available data were used to predict logK(1) values for Sm(III) and Ho(III) with MDP. The logK(1) values for Sm(III)-MDP and Ho(III)-MDP were estimated to be 9.65+/-0.10 and 9.85+/-0.10, respectively. PMID- 16213590 TI - Prevalence of metallo-beta lactamase (MBL) producing Pseudomonas spp. and Acinetobacter spp. in a tertiary care hospital in India. AB - OBJECTIVE: Gram-negative bacteria with acquired metallo-beta lactamases production have been increasingly reported in some countries, necessitating their detection. We determined the occurrence of acquired metallo-beta lactamases (MBL) producing bacteria in our teaching hospital. METHOD: The method employed was a rapid technique using a disk of imipenem plus 750 microg of EDTA, on Mueller Hinton plate which was supplemented with 70 microg/ml of zinc sulfate, to differentiate MBL producing Pseudomonads and Acinetobacter. RESULT: Out of 200 isolates tested, 7.5% of our Pseudomonads and Acinetobacter were shown to be MBL producers. CONCLUSIONS: Though MBL prevalence is still low in our hospital set up, screening for MBL among Pseudomonas and Acinetobacter isolates should be routinely done in laboratory. PMID- 16213591 TI - Seroprevalence of herpes simplex virus 1 and 2 and correlates of infection in Israel. AB - BACKGROUND: The prevalence of infections with herpes viruses 1 and 2 (HSV1 and HSV2) have significantly changed in the developed countries during the last three decades. This study provides up-to-date estimates of HSV-1 and HSV-2 seroprevalence and information on the correlates of infection in a representative sample of the Israeli population. METHODS: Three thousand eight hundred and eighty-one sera collected in 2000-2001 from an age-stratified general population sample were tested using an indirect IgG ELISA for type-specific HSV-1 and HSV-2 antibodies. RESULTS: Unadjusted HSV-1 seroprevalence was 59.8% (95% confidence interval: 58.2-61.4) and increased with age in both genders. Multivariate analysis revealed that HSV-1 seroprevalence rates were significantly higher in females, non-Jews, subjects who were not born in Israel, inhabitants of Jerusalem and Southern Israel and among subjects also seropositive for HSV-2. Unadjusted HSV-2 seroprevalence among adults was 9.2% (95% confidence interval: 7.9-10.2), and was higher in females, peaking at 20.5% in females aged 40-49. In a multivariate model, females, subjects who were not born in Israel and residents of Jerusalem and Southern Israel were significantly associated with HSV-2 seropositivity. CONCLUSIONS: HSV-1 seroepidemiology in Israel resembles the trends in industrialized countries. HSV-2 seroprevalence rates is in the lower range reported across Europe. PMID- 16213592 TI - What is subacute necrotizing fasciitis? A proposed clinical diagnostic criteria. AB - OBJECTIVE: Subacute necrotizing fasciitis is a poorly defined clinical entity. Its very existence has been the subject of much controversy. While rarely reported, subacute forms of necrotizing fasciitis have been documented in the literature by many authors. This paper highlights some recently reported cases in the literature that clearly shows that subacute forms of necrotizing fasciitis indeed exist and may in fact be under-reported because of the lack of awareness and a consistent diagnostic criteria. METHODS: A Medline search was performed with the following keyword; necrotizing fasciitis, subacute, variant and indolent. RESULTS: Majority of reported cases did not give sufficient information to satisfy the reviewer that these cases were indeed subacute forms of necrotizing fasciitis. We identified three cases of subacute necrotizing fasciitis that clearly are subacute cases and analysed their clinical presentation. A diagnostic criterion for defining subacute necrotizing fasciitis was proposed based on these cases and the authors' clinical experiences. CONCLUSION: This proposed diagnostic criterion serves to facilitate future reporting and documentation of this condition. The clinical significance and implication of this are discussed. PMID- 16213593 TI - On the scent of human olfactory orbitofrontal cortex: meta-analysis and comparison to non-human primates. AB - It is widely accepted that the orbitofrontal cortex (OFC) represents the main neocortical target of primary olfactory cortex. In non-human primates, the olfactory neocortex is situated along the basal surface of the caudal frontal lobes, encompassing agranular and dysgranular OFC medially and agranular insula laterally, where this latter structure wraps onto the posterior orbital surface. Direct afferent inputs arrive from most primary olfactory areas, including piriform cortex, amygdala, and entorhinal cortex, in the absence of an obligatory thalamic relay. While such findings are almost exclusively derived from animal data, recent cytoarchitectonic studies indicate a close anatomical correspondence between non-human primate and human OFC. Given this cross-species conservation of structure, it has generally been presumed that the olfactory projection area in human OFC occupies the same posterior portions of OFC as seen in non-human primates. This review questions this assumption by providing a critical survey of the localization of primate and human olfactory neocortex. Based on a meta analysis of human functional neuroimaging studies, the region of human OFC showing the greatest olfactory responsivity appears substantially rostral and in a different cytoarchitectural area than the orbital olfactory regions as defined in the monkey. While this anatomical discrepancy may principally arise from methodological differences across species, these results have implications for the interpretation of prior human lesion and neuroimaging studies and suggest constraints upon functional extrapolations from animal data. PMID- 16213594 TI - MAOI efficacy and safety in advanced stage treatment-resistant depression--a retrospective study. AB - OBJECTIVE: Evidence-based data suggest that MAOI therapy may be effective in up to 50% of patients with treatment-resistant depression (TRD). We hypothesized that MAOI therapy is similarly effective in patients with advanced stage TRD who are unresponsive to > or = 4 prior antidepressant drug (AD) trials compared to patients with early stage TRD who are unresponsive to < or = 3 prior AD trials. METHODS: To test this hypothesis, data were harvested from 400 patient charts. Of these, 59 patients received a total of 75 MAOI treatment trials. 50 patients had 1 MAOI trial and 9 patients had 2 or more MAOI trials. Response was assessed using the Clinical Global Impressions Change (CGI/C) scale. RESULTS: 56% of MAOI trials resulted in a CGI/C score of 1 ("very much better") or 2 ("much better"). Only 25% resulted in a CGI/C score of 4 or more ("no change" or "worse"). 32.5% of MAOI trials resulted in a CGI/C score of 1 in patients with early stage TRD, while only 12.1% of MAOI trials resulted in a CGI/C score of 1 in patients with advanced stage TRD (p=0.04). There was a significant negative correlation between the number of prior, failed AD trials and the final CGI/C score (p=0.03). The odds associated with attaining a CGI/C score of 1 diminished by a factor of 30% with each prior failed AD trial. We observed only 1 case of acute hypertension which responded to sublingual nifedipine therapy. LIMITATIONS: The sample size was limited, and MAOI outcome was not compared with other AD therapy. The adequacy of prior AD trials could not always be verified. CONCLUSION: These data suggest that MAOI therapy may be beneficial in patients with early stage TRD who are unresponsive to < or = 3 prior treatments. However, the relative efficacy of MAOI therapy in advanced stage TRD remains uncertain. PMID- 16213595 TI - Prepartum chronic ultramild stress increases corticosterone and estradiol levels in gestating mice: implications for postpartum depressive disorders. AB - Fluctuations in steroid hormones (glucocorticoids and estradiol) levels during pregnancy and after delivery are thought to contribute to the etiology of postpartum depression. Such changes may be exacerbated by stressful events, which constitute a predisposing factor for postpartum mood disorders. In the present study, blood hormonal variations associated with prepartum Chronic Ultramild Stress (CUMS) exposure were assessed at two times (15th day of pregnancy and 3rd day postpartum) in mice stressed from day 1 of pregnancy to termination of pregnancy. Litter weight and litter size were determined in both groups whereas the duration of pregnancy was determined in the 3-day postpartum group. CUMS increased estradiol and corticosterone levels during pregnancy, but such effects were no longer observed in the postpartum period, where cortisol levels were decreased in control and stressed mice and estradiol levels were reduced in previously stressed mothers. No effects of the CUMS procedure were observed on gestational parameters. Given the link between hormonal variations during pregnancy and subsequent postpartum depression, these results suggest that CUMS applied to gestating female may provide a useful model for the study of the mechanisms of stress, which may lead to postpartum mood disorders. PMID- 16213596 TI - Subjective experiences and the psychopathological dimensions in schizophrenia. AB - The aim of this article was to explore the relationships between subjective experience (SE) and objective symptoms in schizophrenia. Exploratory principal component analysis was used to determine the latent organization of the variables. A group of 310 chronic patients with schizophrenia who met ICD-10 criteria for F20 schizophrenic disorder were included in the study. SE and objective symptoms were rated using, respectively, the 24-item version of the Frankfurt Complaint Questionnaire (FCQ-24) and the Positive and Negative Syndrome Scale (PANSS). An exploratory principal component analysis was performed on the correlation matrix comprising items from both the FCQ-24 and the PANSS. Using several guidelines to select the number of factors, the exploratory principal component analysis yielded a six-factor solution with no overlap of the significant factor loadings for the items from each scale. These six factors represent, respectively, the subjective, negative, positive, disorganization, hostility and anxious-depressive dimensions of the schizophrenic phenomenology. The findings support the view that SE is a construct that is separate and distinct from the objective symptomatology in schizophrenia. PMID- 16213597 TI - Extract from Agaricus blazei Murill can enhance immune responses elicited by DNA vaccine against foot-and-mouth disease. AB - The fungus Agaricus blazei Murill (ABM) is particularly rich in polysaccharides, which have shown particularly strong results in treating and preventing cancers. The goal of this study was to investigate whether co-administration of the ABM extract with foot-and-mouth disease virus (FMDV) DNA vaccine could increase the immune responses. Compared with the control mice, which received FMDV DNA vaccine alone, significant increase in not only the FMDV-specific antibody response but also T cell proliferation was observed in mice which received FMDV DNA vaccine plus the ABM extract. Taken together, these results demonstrated that application of the ABM extract might provide a strategy to improve the efficacy of DNA vaccines. PMID- 16213598 TI - MuSK antibody positive myasthenia gravis plasma modifies MURF-1 expression in C2C12 cultures and mouse muscle in vivo. AB - MG is an antibody-mediated disease that is often treated with corticosteroids. Antibodies to the muscle specific tyrosine kinase (MuSK) have been identified in a proportion of patients with myasthenia gravis (MG) without acetylcholine receptor (AChR) antibodies. MuSK-MG patients often suffer from marked facial muscle weakness, and some patients develop facial and tongue muscle atrophy. MuSK is a receptor tyrosine kinase that plays an essential role during development and is thought to play a trophic role in mature muscle. It is possible, therefore, that the muscle atrophy results from the action of the MuSK antibodies themselves, but effects of corticosteroids on muscle might also be involved. Muscle atrophy in vivo is associated with upregulation of striated Muscle RING Finger protein-1 (MURF-1), and MURF-1 is also upregulated in C2C12 myotubes exposed to the corticosteroid, dexamethasone (Dex). Here we investigated the effects of MuSK antibodies or Dex on MURF-1 expression in C2C12 cultures and in mouse muscles after treatment in vivo, using quantitative Western blotting. We also looked at expression of neural cell adhesion molecule (NCAM, CD56) that is upregulated after denervation in vivo. MuSK-MG plasma and purified IgG from a patient with marked muscle atrophy modestly increased MURF-1 expression in C2C12 cells in culture, and MURF-1 expression in mouse masseter (facial) muscle, but not in gastrocnemius (leg). Dex had a more marked effect on MURF-1 expression in C2C12 cells, but did not affect MURF-1 expression in either muscle. However, both in C2C12 cells and in vivo, Dex substantially reduced NCAM expression. These results provide the first evidence that MuSK-MG plasma can influence expression of an atrophy-related protein, and preliminary evidence that a facial muscle, the masseter, is more susceptible to this effect. They indicate the need for further studies on muscle atrophy, MuSK-MG antibodies, the effects of steroids, and the intracellular pathways involved. PMID- 16213599 TI - Communication disorders in Nigerian children. AB - BACKGROUND/AIM: Communication disorders have been acknowledged as a major public health issue because they compromise early childhood development, restrict vocational attainment and undermine the economic well being of the society. The aim of this study is to determine the pattern of communication disorders among children in a developing country and the requisite intervention services. MATERIALS AND METHODS: This prospective study was conducted in Lagos University Teaching Hospital, Lagos between January 2002 and June 2003 among children aged 6 months to 15 years that presented in the audiology clinic of the hospital with communication disorders. All the patients had neurological, otolaryngological, audiological and speech evaluations. RESULTS: A total of 184 patients were seen during the period out of whom 136 (74%) were between the ages of 6-47 months. Hearing impairment was documented in 120 (65.2%) children, speech disorders in 56 (30.4%), rhinolalia 2.2% and stuttering 2.2%. Of those with hearing impairment, 70% had delayed speech and language. Among children with speech disorders 78.6% had specific language impairment (SLI). Aetiological factors recorded for the communication disorders were seizures 10.9%, measles 8.7% meningitis 8.7%, birth asphyxia 6.5%, otitis media with effusion (OME) 4.3%, kernicterus 4.3%, congenital deformity 4.3%, ototoxicity 2.2%, cerebral palsy 2.2%, and undetermined causes 47.9%. CONCLUSION: Hearing impairment is the commonest communication disorder. Early detection and appropriate follow up is recommended for all children in their first year of life. The role of parents and caregivers in seeking early help should be strengthened while capacity building for the training of more audiologists and speech therapists should be pursued rapidly. PMID- 16213600 TI - Secreted expression of the classical swine fever virus glycoprotein E(rns) in yeast and application to a sandwich blocking ELISA. AB - E(rns) is an envelope glycoprotein of classical swine fever virus (CSFV) with RNase activity. The purpose of this study was to produce an active E(rns) for further applications using the yeast secreted expression system. The E(rns) gene was cloned into the expression vector pGAPZalphaC which was introduced into Pichia pastoris. Expression of E(rns) protein in culture supernatant was confirmed by Western blot analysis using both the monoclonal antibody against CSFV E(rns) and CSFV-positive swine serum. The yeast-expressed E(rns) (yE(rns)) was shown to have N-linked glycosylation and to form homodimer of 74 kDa molecules. All monomer, homodimer, and deglycosylated forms of yE(rns) demonstrated intrinsic ribonuclease activity and a clear preference for uridine rich sequence. A direct sandwich blocking enzyme-linked immunosorbent assay (ELISA) based on the yE(rns) was developed with a high sensitivity and specificity. The yE(rns) which possesses enzymatic activity and retains antigenicity may provide a useful material for developing a diagnostic kit. PMID- 16213601 TI - A rapid method for estimation of baculovirus titer based on viable cell size. AB - Baculovirus protein expression system is a powerful tool for producing recombinant proteins. To optimize conditions for efficient recombinant protein expression, it is important to determine titer of virus stock for arriving at an optimal multiplicity of infection (MOI) that maximizes recombinant protein expression. Traditionally plaque assays have been used for titer determination. Other methods such as endpoint dilution, quantitative real-time polymerase chain reaction and flow cytometry have been developed to aid the determination of virus titers. However, most of these methods are time-consuming and labor intensive. In this regard, a simple and rapid method for determination of virus titers based on the cytopathic effects that lead to viable cell size increase following virus infection is presented in this paper. In this study, the Vi-CELL (Beckman Coulter) was used to measure cell-diameter over a range of virus dilutions, following infection. Applying statistical modeling techniques, the viable cell diameter data was used to estimate the virus titer. The results indicated that the viable cell-diameter based titer estimation to be reliable and comparable to titers determined by the traditional plaque assay. PMID- 16213602 TI - Stem cells for the treatment of myelin loss. AB - Treatment of myelin loss is particularly suited to therapeutic strategies based on cell replacement. Demyelination represents a defined and functionally debilitating deficit, and remyelination can be accomplished by supplying regions of demyelination with myelinogenic cell populations. Clinical interest in stem cells as a source of myelinogenic cells arises from their ability to provide an apparently unlimited cell supply for transplantation, and from recent demonstrations that they can be directed to myelinogenic phenotypes with high purity. Here, I present the emerging perspective that stem-cell-mediated remyelination of the adult CNS is a viable therapeutic strategy, and discuss the challenges to remyelination posed by the environment of acute and chronic injuries. PMID- 16213603 TI - Temporary inactivation of the medial and basolateral amygdala differentially affects TMT-induced fear behavior in rats. AB - Trimethylthiazoline (TMT) is a component of fox feces and is thought to be a stimulus with innate fear-eliciting properties for rodents. Naive laboratory rats that are exposed to TMT display freezing behavior, a known behavioral sign of fear and anxiety. Early studies examining the neural basis of TMT-induced fear showed that the bed nucleus of the stria terminalis is important for this behavior. In contrast, the central and lateral nuclei of the amygdala does not seem to participate in the neural processing of TMT-induced fear. However, a study investigating c-fos expression in response to TMT-exposure revealed a strong activation of the medial as well as a weak activation of the basolateral amygdala. Therefore, the present study examined the effects of temporary inactivation of the medial and basolateral amygdala on TMT-induced freezing. Temporary inactivation was accomplished by local injections of the GABA(A) receptor agonist muscimol into the areas of interest. TMT-induced freezing was completely blocked by temporary inactivation of the medial amygdala. Temporary inactivation of the basolateral amygdala resulted in a delay of the onset of the freezing response to TMT. These results clearly demonstrate that the medial amygdala is crucial for TMT-induced freezing, whereas the basolateral amygdala seems to play a modulatory role in this type of fear behavior. Since the medial amygdala is also involved in the processing of cat odor-induced fear, the finding of the present study points towards a general role of the medial amygdala in the processing of predator odor-induced fear. PMID- 16213604 TI - Differential metamorphosis alters the endocrine response in anuran larvae exposed to T3 and atrazine. AB - Pesticide chemical contamination is one of the suspected contributors of the amphibian population decline. The herbicide atrazine is one of the major surface water contaminants in the U.S. A previous study has shown that atrazine at concentrations as low as 100 parts per billion (ppb) increased the time to metamorphosis in Xenopus laevis tadpoles. However, questions remain as to the applicability of a study of a non-native species to a native organism. The possible effects of atrazine on developing Bufo americanus were explored. Atrazine at potentially (albeit high) environmental concentrations was found not to delay the metamorphosis of developing B. americanus tadpoles as observed in X. laevis. Several studies have indicated that atrazine affects thyroid hormones. Since thyroid hormones are critical in amphibian metamorphosis, B. americanus and X. laevis tadpoles were exposed to exogenous 3,5,3'-triiodothyronine (T3). X. laevis were found to be more responsive to the effects of exogenous T3 compared to B. americanus, indicating that X. laevis may be more sensitive to endocrine active chemicals than B. americanus. In X. laevis, nuclear heterogeneity has been associated with metamorphosis. Flow cytometric analysis of the nuclei of normal metamorphing B. americanus indicates a decrease in the amount of thyroid mediated chromatin alterations relative to the nuclei of metamorphing X. laevis. Indications are that the differential response to endocrine disruption is due to the differential role of chromatin associated gene expression during metamorphosis of B. americanus versus X. laevis. A second native species, Hyla versicolor, was observed to have the X. laevis nuclear pattern with respect to metamorphosis. As such, sensitivity to endocrine disruption is hypothesized not to be limited to laboratory non-native species. PMID- 16213605 TI - Organochlorine compounds in liver and concentrations of vitellogenin and 17beta estradiol in plasma of sea bass fed with a commercial or with a natural diet. AB - Results from previous experiments directed to determine the effect of different nutritional factors or the effect of xenobiotics on hormonal control of reproduction, lead to the hypothesis that hormonal perturbations repeatedly observed in sea bass (Dicentrarchus labrax) broodstock feeding commercial diets could have been caused by the presence of aryl hydrocarbon receptor (AhR) ligands, such as dioxins, furans and polychlorinated biphenyls (PCBs) in the diet. To evaluate this hypothesis, dioxins and related compounds were analysed in liver of female sea bass fed with a commercial or with a natural diet consisting of trash fish (bogue, Boops boops), and concentrations of vitellogenin (VTG) and 17beta-estradiol (E2) were determined in plasma obtained previously in monthly samplings of these animals. As observed in other experiments, females fed with a commercial diet exhibited lower VTG and higher E2 plasma levels than females fed with the natural diet. In liver, sea bass fed with the commercial diet exhibited a profile clearly dominated by high-chlorinated dioxins while in fish fed with the natural diet this profile was dominated by low chlorinated furans. However, typical AhR ligands, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin showed no differences between groups or, as is the case of planar PCBs, showed higher concentrations in the liver of fish fed with the natural diet. These results do not permit to explain the observed hormonal alterations by a possible antiestrogenic effect caused by dioxins and related compounds. PMID- 16213606 TI - Cardiac effects of oxytocin: is there a role for this peptide in cardiovascular homeostasis? AB - Oxytocin is well known for its role in reproduction. However, evidence has emerged suggesting a role in cardiovascular and hydroelectrolytic homeostasis. Although its renal effects have been characterized, the cardiac ones have not been much studied. Therefore, we aimed to investigate the cardiac effects of oxytocin both in vivo and in vitro. In unanesthetized rats (n=6) intravenous oxytocin (1 mug) decreased dP/dt(max) by 15% (P<0.05) and heart rate by 20% (P<0.001), at the first minute after injection. dP/dt(max) was still lower in OT treated rats than in controls (n=8) after 15 min (P<0.05), while heart rate returned to control values after 5 min. In isolated hearts, oxytocin was able to promote negative inotropic and chronotropic effects. Perfusion with 10(-5), 10( 6) and 10(-7)M oxytocin resulted in approximately 60% (P<0.01), 25% (P<0.01) and 10% (P<0.05) reduction of left ventricle developed pressure, without effect in lower concentrations (10(-10) to 10(-8) M). Also, dP/dt(max) was reduced by 45 and 20% (10(-5) e 10(-6) M; P<0.01), while diastolic pressure raised and heart rate fell only with 10(-5)M oxytocin (P<0.05). Intravenous oxytocin (1 mug; n=6) increased arterial pressure by 22% at the first minute (+23+/-3 mm Hg; P<0.001), returning to control value thereafter. Thus, oxytocin is able to promote directly negative inotropic and chronotropic effects, but its in vivo effect also involves a reflex mechanism, originated from its pressor effect. PMID- 16213607 TI - Time-related distribution, risk factors and prognostic influence of embolism in patients with left-sided infective endocarditis. AB - BACKGROUND: Few studies evaluated systemic arterial embolism after beginning of symptoms of infective endocarditis in a large series of patients. METHODS: We studied 629 patients with left-sided infective endocarditis, aged 37.9+/-17.3 years, 396(63%) men and 233(37%) women. Endocarditis occurred on native valves in 405(64.4%) patients and on prosthetic heart valves in 224(35.6%). Infecting microorganisms were streptococci in 297(47.3%) patients, Staphylococcus aureus in 77(12.3%), Staphylococcus epidermidis in 56(8.9%), enterococci in 51(8.1%), Gram negative bacteria in 33(5.2%), fungi in 9(1.4%) and other microorganisms in 27(4.2%). In 79(12.6%) patients blood cultures were negative. RESULTS: 146 embolic events occurred in 133(21.1%) out of 629 patients; in 63(47.4%) of them emboli affected the central nervous system, in 57(42.9%) affected peripheral organs and in 13(9.7%) affected both the central nervous system and peripheral organs. Embolism occurred between beginning of symptoms of endocarditis and antimicrobial therapy in 56(42.1%) patients and on the day therapy started in 18(13.5%); 109(81.9%) embolic events occurred up to the 15th day of antimicrobial therapy. Embolic risk was higher in S. aureus endocarditis (relative risk 2.97); in patients with a mitral (relative risk 2.4) or aortic (relative risk 3.3) prosthetic valve and vegetations on echocardiography. Embolic risk was lower in patients with a longer duration of symptoms. The death risk doubled in patients with embolism (relative risk 2.01). CONCLUSIONS: Embolic events were more frequently early events after beginning of symptoms of infective endocarditis. Embolic risk was higher in S. aureus endocarditis and in patients with prosthetic heart valves and vegetations on echocardiography. PMID- 16213608 TI - Independent association between triglycerides and coronary artery disease in Taiwanese type 2 diabetic patients. AB - BACKGROUND: This study evaluated the association between triglycerides (TG) and coronary artery disease (CAD) in Taiwanese adults with type 2 diabetes mellitus (T2DM). METHODS: A total of 1150 patients (542 men and 608 women) aged 62.5+/ 11.6 years were studied. CAD was diagnosed by history or an abnormal electrocardiogram (coronary probable or possible by Minnesota codes). Age, body mass index (BMI), smoking, use of insulin, anti-hypertensive agents and lipid lowering agents, fasting plasma glucose (FPG), hemoglobin A1c (HbA1c), systolic blood pressure (SBP), diastolic blood pressure (DBP), serum total cholesterol (TC), high-density lipoprotein cholesterol (HDL-c) and low-density lipoprotein cholesterol (LDL-c) were considered as potential confounders. RESULTS: Patients with CAD were older, had higher prevalences of use of anti-hypertensive and lipid lowering agents, and had higher BMI, SBP, DBP and TG. CAD increased significantly with increasing quartiles of TG (P-trend < 0.001). Ln(TG) was significantly correlated with BMI, FPG, HbA1c, DBP, TC, HDL-c (inversely) and LDL-c. Ln(TG) was associated with CAD with an unadjusted odds ratio of 1.411 (1.145-1.740). The odds ratio after adjustment for all confounders was slightly attenuated but still statistically significant: 1.380 (1.043-1.826). None of the other lipid parameters of TC, HDL-c and LDL-c were significantly associated with CAD in logistic models when they were entered for adjustment either separately or simultaneously. Sensitivity analyses by using history alone or history and coronary probable as diagnostic criteria for CAD did not change the association between TG and CAD. CONCLUSIONS: TG is an independent risk factor for CAD in Taiwanese T2DM, independent of TC, HDL-c, LDL-c or other confounders. PMID- 16213609 TI - Neurohormonal activation and diagnostic value of cardiac peptides in patients with suspected mild heart failure. AB - BACKGROUND: Data describing activation of brain natriuretic peptide (BNP) system relative to the renin-angiotensin-aldosterone system (RAAS) are sparse in the early phase of heart failure (HF). AIMS: To compare activation of BNP system relative to RAAS hyperactivity and to assess diagnostic accuracy of cardiac peptides to detect any left ventricular dysfunction (LVD) in patients referred from primary care with suspected HF before institution of medical therapy. METHODS: Of 166 referred patients 150 were consecutively included (14 were excluded and two refused consent). Echocardiography and measurements of neurohormonal activity were performed. Systolic dysfunction (LVSD) was defined as an ejection fraction150 bovine viral diarrhea (BVD) viruses isolated from cattle in Switzerland belonged to genotype 1, with subgenogroups e, h, k and b found in decreasing frequency. To date, representatives of subgenogroup k have been detected in Switzerland only. Despite serological evidence of Border disease in sheep, only few Border disease viruses have been isolated, all of which belong to the novel group 3. Serological evidence suggested that pestivirus infections may occur also in wild ruminants in Switzerland but no isolates are available for analysis. In addition, we describe two pestiviruses, one a cell culture contaminant and the other isolated from a buffalo, that cluster with a recently proposed novel pestivirus species. PMID- 16213617 TI - Severity of illness and priority setting: worrisome lack of discussion of surprising finding. PMID- 16213618 TI - Time series observation based InfraRed Epifluorescence Microscopic (TIREM) approach for accurate enumeration of bacteriochlorophyll-containing microbes in marine environments. AB - Bacteriochlorophyll a Containing Microbes (BCM) are a unique group of microorganisms in the marine environment. Accurate determination of their abundance is critical for understanding their role in energy flow and carbon cycle in the ecosystem. The InfraRed Epifluorescence Microscopy (IREM) method, using infrared fluorescence as the diagnostic signal of BCM, is the most convenient means to date for enumeration of BCM in seawater, but IREM methodology suffers from serious errors introduced by cyanobacteria, which also can emit infrared fluorescence and whose abundance is of the same order of magnitude as BCM. In the present study, an advanced "Time-series observation based cyanobacteria-calibrated InfraRed Epifluorescence Microscopy (TIREM)" approach is established for accurate enumeration of BCM in marine environments. The protocol is distinguished by its use of time series observation, auto-imaging and digital analysis. In principle, the correct count of BCM can be obtained by subtracting the cyanobacterial count from the total infrared positive count. The challenge, however, is that Prochlorococcus, the most abundant cyanobacterium in the sea, is readily visible in infrared images but not visible in the initial cyanobacterial images obtained by epifluorescence microscopy because its emission signals are masked by brighter fluorescence from larger cells like Synechococcus coexisting in seawater samples. Prochlorococcus cells become gradually visible when the fluorescence from Synechococcus cells declines after a period of exposure to excitation light. Therefore the plateau (maximum) count of the cyanobacterial cells in time series images rather than in the initial ones, as previously believed, represents the correct count for the total number of cyanobacteria (Synechococcus plus Prochlorococcus cells). Thus, the accurate estimation of BCM abundance can only be calculated from the formula: [BCM cells] = [plateau count of infrared positive cells]-[plateau count of cyanobacterial cells]. The conceptual advance of the TIREM protocol is that in classical epifluorescence microscopy or in IREM protocols, quick observation is recommended to avoid quenching the fluorescence, but in the TIREM protocol, instead, time series observation is the key for obtaining reliable data. The TIREM protocol is validated by studies using BCM and cyanobacterial pure cultures as well as by examination of samples from various marine environments. PMID- 16213620 TI - Plasma homocysteine levels are not increased in murine models of Alzheimer's disease. AB - Alzheimer's disease (AD) patients tend to have increased plasma levels of homocysteine. However, it is unclear whether abnormality in homocysteine levels is a primary cause of Alzheimer's disease or a disease marker. In order to investigate the relative impact of Alzheimer's disease on plasma homocysteine levels, total plasma homocysteine levels were evaluated in transgenic mouse models that exhibit abnormalities in their brains that are similar to Alzheimer's patients. No significant difference was observed in blood of murine models compared to control mice, indicating that elevated plasma homocysteine level seems to be a risk marker at the most. PMID- 16213619 TI - Cancer in the oropharynx: cost calculation of different treatment modalities for controlled primaries, relapses and grade III/IV complications. AB - BACKGROUND AND PURPOSE: This paper presents a model for cost calculation using the different treatment modalities for oropharyngeal (OPh) cancers used in our hospital. We compared full hospital costs, the associated costs of localregional relapses (LRR) and/or treatment related grade III/IV complications. MATERIALS AND METHODS: Patients with OPh cancer are treated in the Erasmus MC preferably by an organ function preservation protocol. That is, by external beam radiation therapy (EBRT) followed by a brachytherapy (BT) boost, and neck dissection in case of N+ disease (BT-group: 157 patients). If BT is not feasible, resection with postoperative EBRT (S-group [S=Surgery]: 110 patients) or EBRT-alone (EBRT-group: 77 patients) is being pursued. Actuarial localregional control (LRC), disease free survival (DFS) and overall survival (OS) at 5-years were calculated according to the Kaplan-Meier method. The mean costs per treatment group for diagnosis, primary Tx per se, follow-up, (salvage of) locoregional relapse (LRR), distant metastasis (DM), and/or grade III/IV complications needing clinical admission, were computed. RESULTS: For the BT-, S-, or EBRT treatment groups, LRC rates at 5-years were 85, 82, and 55%, for the DFS, 61, 48, and 43%, and for the OS 65, 52, and 40%, respectively. The mean costs of primary Tx in case of the BT group is 13,466; for the S-group 24,219, and 12,502 for the EBRT-group. The mean costs of S (the main salvage modality) for a LRR of the BT group or EBRT-group, were 17,861 and 15,887, respectively. The mean costs of clinical management of Grade III/IV complications were 7184 (BT-group), 16,675 (S-group) and 6437 (EBRT group). CONCLUSION: The clinical outcome illustrates excellent LRC rates at 5 years for BT (85%), as well as for S (82%). The relatively low 55% LRC rate at 5 years for EBRT probably reflects a negative selection of patients. It is of interest that the total mean costs of patients alive with no evidence of disease is least for the BT-group: 15,101 as opposed to 25,288 (S) and 18,674 (EBRT). Main underlying cause for the high costs with S as opposed to RT alone is the number of associated clinical admission days, not only during primary treatment, but also at relapse. This might be taken into consideration when treating these patients. PMID- 16213621 TI - Effects of water activity and temperature on germination and growth profiles of ochratoxigenic Penicillium verrucosum isolates on barley meal extract agar. AB - The aim of this work was to study the combined effects of water activity (aw, 0.75-0.99), temperature (10-30 degrees C) and isolate on spore germination and mycelial growth of ochratoxigenic Penicillium verrucosum isolates on barley meal extract agar (BMEA). Lag phases prior to germination (h), germination rates (h( 1)) and growth rates (mm day(-1)) were obtained at each set of conditions and for all the isolates. Analysis of variance showed a significant influence of the abiotic factors assayed on fungal development. Minimum aw levels found for germination and mycelial growth were 0.80 and 0.85, respectively, while maximum germination and growth rates were obtained at 0.95-0.99 aw level. Although maximum germination was obtained at 20 degrees C, significant differences were not found between growth rates at 20 and 30 degrees C. Data were modelled by MLR regressions and response surface models were obtained. Spore germination and mycelial growth of ochratoxigenic P. verrucosum isolates are significantly affected by water activity and temperature. Moreover, different isolates may have slightly different response to environmental factors. Predictive models obtained may allow a rough prediction of germination/growth as a function of the storage temperature and moisture content of barley grains. PMID- 16213622 TI - Effects of antimicrobial components of essential oils on growth of Bacillus cereus INRA L2104 in and the sensory qualities of carrot broth. AB - The possible use of antimicrobials from seven plant essential oils as food preservatives was studied by examining their effects on the growth kinetics of activated Bacillus cereus INRA L2104 spores inoculated into tyndallized carrot broth. The effects of various concentrations of borneol, carvacrol, cinnamaldehyde, eugenol, menthol, thymol, and vanillin were determined. Five microliters of cinnamaldehyde, 15 microl of carvacrol, or 30 mg of thymol per 100 ml of inoculated carrot broth completely inhibited bacterial growth for more than 60 days at 16 degrees C. Lower concentrations of the three antimicrobials prolonged the lag phase and reduced both the exponential growth rate and the final population densities of cultures. The study of the sensory characteristics of the supplemented broths suggested that low concentration of cinnamaldehyde enhanced the taste of carrot broth, and that it did not have any adverse effect on the taste and smell of carrot broth at concentrations less than 6 microl 100 ml(-1). PMID- 16213623 TI - ITS primers for the identification of marketable Boletus. AB - Boletus species belonging to the section Boletus are the most frequently eaten fungi among those harvested in natural conditions in Europe. This section groups 10 taxa which are hardly distinguishable on the basis of their morphology. Some of them have been shown to induce allergic IgE-mediated symptoms either through inhalation, ingestion or contact. Since questions relating to the presence of allergens in any of the species most in demand (B. edulis, B. aereus, B. pinophilus, B. aestivalis, all classified as B. edulis s.l.) remain open, together with the absence of tools which distinguish the species, we sequenced the ITS region of 28 Boletus samples and then we designed specific primers. These allowed the effective separation of the taxa. In addition, the phylogenetic tree obtained from the sequences alignment revealed that B. violaceofuscus, a spectacular Chinese fungus considered belonging to the section Boletus and often sold intermixed with B. edulis s.l. specimens, clusters outside the section Boletus. PMID- 16213624 TI - "Yes", "No" or "Yes, but"? Multinomial modelling of NICE decision-making. AB - The National Institute for Health and Clinical Excellence (NICE) issues mandatory guidance on health technologies to the UK NHS, based on clinical evidence, cost effectiveness and other considerations. However, the exact factors considered, their relative importance and tradeoffs between them are not made explicit. Previous research modelled NICE decisions as a binary choice (accept/reject) dependent on cost-effectiveness, amongst other variables. This paper proposes and tests an alternative model of decision-making that may better represent the "yes, but..." nature of many NICE decisions. Decisions were categorised as "recommended for routine use", "recommended for restricted use" or "not recommended". The NICE appraisal process was modelled as a single decision between the three categories. Multinomial logistic regression techniques were used to evaluate the impact of: quantity/quality of clinical evidence; cost-effectiveness; decision date; existence of alternative treatments; budget impact; technology type. Results suggest that interventions supported by more randomised trials are more likely to be recommended and endorsed for routine use. Higher cost-effectiveness ratios increased the likelihood of interventions being rejected rather than recommended for restricted use but did not significantly affect the decision between routine and restricted use. Pharmaceuticals, interventions appraised early in the NICE programme and those with more systematic reviews were also less likely to be rejected, while patient group submissions made a recommendation for routine rather than restricted use more likely. The presence of factors affecting the decision between routine and restricted use but not that between routine use and rejection suggests that modelling these three outcomes reflects NICE decision making more closely than binary-choice analyses. PMID- 16213625 TI - Clonal spread of meropenem-resistant Acinetobacter baumannii strains in hospitals in the Mediterranean region and transmission to South-west Germany. PMID- 16213626 TI - Protective environments and health status: cross-talk between human and animal studies. AB - Although aging populations tend to have increased prevalence of a diversity of diseases and disabilities, there are substantial numbers of people who, nevertheless, maintain good health into old age. Human studies frequently demonstrate associations between environmental factors, particularly supportive social environments, and positive states of health. Identifying the pathways from protective social environments to reduced disease risk necessitates the use of animal models as a basis of explanation and a source of suggestions for further human research. We present two examples of this kind of cross-talk: (i) the possibility that the success of well-being therapy following pharmacological treatment for depression as a means of preventing recurrent depressive episodes is based on the stimulation of enrichment of dendritic networks in the hippocampus and spine retraction in the basolateral amygdala; (ii) the possibility that the release of intracerebral oxytocin is a mediating factor between persistently supportive social environments and reduced disease in later life, as exemplified by low levels of allostatic load. PMID- 16213627 TI - Diabetes, aging, and cognitive decline. AB - Type 1 diabetes is associated with cognitive changes in children and adults, but the extent to which cognition declines with increasing age, and increasing duration of diabetes, remains poorly understood. This cross-sectional study assessed neuropsychological performance on 200 diabetic and 175 nondiabetic adults, 18-64 years of age, stratified into five age bands. Similar age-related cognitive declines were seen on measures of problem-solving, learning and memory, and psychomotor speed, but it was only on the latter measure that diabetic and nondiabetic subjects differed significantly. The best predictor of psychomotor slowing was the presence of clinically significant biomedical complications, particularly proliferative retinopathy, peripheral neuropathy, and peripheral vascular disease (PVD). It now appears that psychomotor slowing is the fundamental cognitive deficit associated with diabetes mellitus; why other cognitive skills are relatively unaffected remains poorly understood. PMID- 16213628 TI - Parkin-mediated lysine 63-linked polyubiquitination: a link to protein inclusions formation in Parkinson's and other conformational diseases? AB - Most, if not all, neurodegenerative diseases are marked by the presence of ubiquitin-positive protein inclusions. How proteins within these inclusion bodies escape proteasomal degradation despite being enriched with ubiquitin remains a conundrum. Current evidence suggests a relationship between proteasomal impairment and inclusion formation, a persuasive explanation for the inability of the cell to remove ubiquitinated protein aggregates. Alternatively, the formation of ubiquitin-enriched inclusion may be uncoupled from the proteasome. Supporting this, we recently uncovered a novel, proteasomal-independent, catalytic activity for the Parkinson disease (PD)-linked ubiquitin ligase, parkin, that significantly enhances the formation of Lewy body (LB)-like inclusions generated in cultured cells by the co-expression of alpha-synuclein and synphilin-1. This unique activity of parkin mediates a non-classical, lysine (K) 63-linked ubiquitin multichain assembly on synphilin-1 that is distinct from the classical, degradation-associated, K48-linked ubiquitination. Interestingly, two other PD linked gene products, alpha-synuclein and UCHL1, have recently also been associated with K63-linked ubiquitination. Inclusive of parkin, there are therefore now three PD-related gene products that are known to potentiate K63 linked ubiquitination, thus signalling an important functional relationship between this unique mode of ubiquitin tagging and PD pathogenesis. Mechanistically, the involvement of a "non-degradative" mode of ubiquitination in protein inclusion formation is an attractive explanation for how proteins are seemingly stabilized within inclusions. PMID- 16213629 TI - Positive affect and biological function in everyday life. AB - There is accumulating evidence that positive affect may protect against ill health and risk of disease. Here, we summarize results from our research program into the biological correlates of positive affect. Data have been collected from middle-aged men and women, with positive affect assessed through repeated ratings of happiness over a working day. The results indicate that greater happiness is associated with lower salivary cortisol both on working and nonworking days, reduced fibrinogen stress responses, and lower ambulatory heart rate in men. These effects are independent of age, socioeconomic status, smoking, body mass and psychological distress. A 3 year follow-up has confirmed these biological associations with happiness. In addition, we found that happiness was inversely related to ambulatory systolic blood pressure on follow-up, again independently of potential confounders including negative affect. These results suggest that positive affective states are linked to favorable health outcomes through their influence on health-related biology, and may be particularly relevant in old age, when the accumulation of risk factors leads to increased risk of chronic disease. PMID- 16213630 TI - Prediction of longitudinal cognitive decline in normal elderly with subjective complaints using electrophysiological imaging. AB - An extensive literature reports changes in quantitative electroencephalogram (QEEG) with aging and a relationship between magnitude of changes and degree of clinical deterioration in progressive dementia. Longitudinal studies have demonstrated QEEG differences between mild cognitively impaired (MCI) elderly who go on to decline and those who do not. This study focuses on normal elderly with subjective cognitive complaints to assess the utility of QEEG in predicting future decline within 7 years. Forty-four normal elderly received extensive clinical, neurocognitive and QEEG examinations at baseline. All study subjects (N = 44) had only subjective complaints but no objective evidence of cognitive deficit (evaluated using the Global Deterioration Scale [GDS] score, GDS stage = 2) at baseline and were re-evaluated during 7-9 year follow-up. Baseline QEEGs of Decliners differed significantly (p < 0.0001, by MANOVA) from Non-Decliners, characterized by increases in theta power, slowing of mean frequency, and changes in covariance among regions, especially on the right hemisphere. Using logistic regression, an R2 of 0.93 (p < 0.001) was obtained between baseline QEEG features and probability of future decline, with an overall predictive accuracy of 90%. These data indicate high sensitivity and specificity for baseline QEEG as a differential predictor of future cognitive state in normal, subjectively impaired elderly. PMID- 16213632 TI - Immunogenicity and safety of two live-attenuated tetravalent dengue vaccine formulations in healthy Australian adults. AB - We conducted a Phase 1b study to evaluate the immunogenicity and safety of two live attenuated tetravalent dengue vaccines in healthy adult volunteers. After one injection, all subjects reported systemic reactions consistent with a mild dengue-like syndrome. Seven volunteers developed dengue 3 viraemia after vaccination. All subjects developed a neutralizing antibody response against serotype 3 with partial response against other serotypes. The trial was stopped early (after 10 subjects enrolled) due to formulation issues, which were related to the dengue 3 vaccine component. Managing viral interference and balancing attenuation to produce acceptable tetravalent immunogenicity with minimal reactogenicity may be a recurring problem for future multivalent live vaccines. PMID- 16213631 TI - Interleukin-12 induces a Th1-like response to Burkholderia mallei and limited protection in BALB/c mice. AB - We evaluated the effect of interleukin (IL)-12 on the immune response to Burkholderia mallei in BALB/c mice. Mice were vaccinated with non-viable B. mallei cells with or without IL-12. There was a seven- to nine-fold increase in IgG2a levels, and a significant increase in the proliferative response and interferon (IFN)-gamma production by splenocytes from mice that received B. mallei and IL-12. We saw an increase in survivors in the groups of mice that received B. mallei and IL-12 when challenged, compared to mice that received only B. mallei or IL-12. The results suggest that IL-12 can enhance the Th1-like immune response to B. mallei and mediate limited protection from a lethal challenge. PMID- 16213634 TI - Radon exhalation from phosphogypsum building boards: symmetry constraints, impermeable boundary conditions and numerical simulation of a test case. AB - Comprehensive understanding of (222)Rn exhalation from phosphogypsum-bearing building material and its accumulation in indoor air is likely to rely on numerical simulation, particularly if transient effects, three-dimensional domains and convection are to be included and investigated. Yet, experimental data and analytical results are helpful (if not crucial) as far as validation is concerned. Having in mind computational code simplicity and in the light of a recent experimental and theoretical report on (222)Rn release from phosphogypsum boards for housing panels, this paper presents and discusses an alternative testing set-up and the corresponding boundary conditions, namely one side of the panel bounded by impermeable wall. Although this is a new facility to be tested, the resultant steady-state one-dimensional diffusion-dominant analytical solution is shown to match the counterpart deduced in the aforementioned previous report, despite it relaxes the constraint of symmetry about the phosphogypsum board centerline, which is inferred in that prior experimental scenario. In addition, numerical results are conducted for a diffusion-dominant two-dimensional time varying test case concerning (222)Rn accumulation in a closed chamber having an exhaling phosphogypsum board tightly placed at one wall. PMID- 16213633 TI - Field trials of a very potent rabies DNA vaccine which induced long lasting virus neutralizing antibodies and protection in dogs in experimental conditions. AB - Kinetics of antibody responses and protection against rabies were investigated after injection of a single dose of rabies DNA vaccine and compared to those induced by one or two injections of cell culture-derived vaccine in dogs issued from the common local breed and reared in experimental conditions. Rabies DNA vaccine administered intradermally by a jet injector in the inner face of the ear was by far more efficient in inducing long lasting high titers of virus neutralizing antibodies compared to cell culture vaccine Rabisin administered either subcutaneously or intramuscularly. Four years after vaccine administration of either DNA or cell culture-derived rabies vaccines, full protection against a rabies peripheral challenge was achieved. Vaccine trials targeting dogs living in field conditions in Tunisia further established that rabies DNA-based vaccination induced a stronger induction of virus neutralizing antibodies compared to Rabisin. This report shows for the first time that DNA vaccination could be more efficient under experimental or field conditions in large size mammals than the best commercially available cell culture-derived vaccine. This improvement will hopefully allow a better rabies control in developing countries by using a more efficient vaccination with fewer doses and targeting all categories of dogs. PMID- 16213635 TI - Distal cervical caries in the mandibular second molar: an indication for the prophylactic removal of the third molar? AB - AIMS: Distal cervical caries (DCC) in mandibular second molar teeth are responsible for the removal of up to 5% of all mandibular third molars. Our aim was to identify the clinical features of these patients. METHODS: We evaluated the records of 100 patients who had 122 mandibular third molars removed because of distal cervical caries in the second molar. RESULTS: Eighty-two percent of third molars had a mesial angulation of between 40 degrees and 80 degrees. The peak age for removal of third molars was 5 years later than in other studies and patients had better dental health than average. The incidence of distal cervical caries DCC has been shown to increase with age. CONCLUSION: Distal cervical caries is a late phenomenon and has been reported only in association with impacted third molars. The early or prophylactic removal of a partially erupted mesio-angular third molar could prevent distal cervical caries forming in the mandibular second molar. PMID- 16213636 TI - The Ilizarov external fixator: what remains of the wire pretension after dynamic loading? AB - BACKGROUND: Maintenance of wire pretension in an Ilizarov external fixator is dependent on the torque applied to the fixation bolts. We therefore measured immediately after surgery the clinically applied torques. The median value was only 10 N m (range 8-14N m). We wondered whether this value is appropriate to maintain the wire pretension and thereby to achieve sufficient fracture stability during dynamic loading of the device for a longer period. METHODS: A material testing machine dynamically loaded one wire mounted on one ring. Several configurations were tested. RESULTS: A quick decrease in wire tension to a steady state situation was seen. In the most stable configuration (20 N m wire fixation torque) 50% of the initial 90 kg wire pretension remained after dynamic loading with 200 N. In the least stable configuration (10 N m torque) considerable wire slippage occurred even without loading and no tension remained after loading! No plastic deformation of the wires was observed so loss of wire tension was due to slippage of the wires through the fixation bolts. INTERPRETATION: With the small fixation torques used in clinical practice considerable wire tension is lost even after a few loading cycles. Further research should address the question whether preservation of a higher wire tension during long term loading promotes faster fracture healing. PMID- 16213637 TI - Removal of cyanobacterial blooms in Taihu Lake using local soils. II. Effective removal of Microcystis aeruginosa using local soils and sediments modified by chitosan. AB - After sepiolite was modified with Fe3+ to increase its surface charge, the initial algal removal rate increased significantly, but its Q8h was not improved substantially at clay loadings below 0.1 g/L. Modification on netting and bridging properties of clays by either chitosan or polyacrylamide (PAM) dramatically increased flocculation (Q8h) of MA cells in freshwaters. Algal removal efficiencies of different solids, including Type III clays, local soils and sediments, were all improved to a similar level of >90% at a total loading of 0.011 g/L (contained 0.001 g/L chitosan) after they were modified with chitosan, making the idea of clearing up algal blooms using local soils/sediments possible. The mechanism of netting and bridging was confirmed to be the most important factor in improving the removal efficiency of cells, whereas clays also played important roles in the sedimentation of the floc. PMID- 16213638 TI - Seasonal development of ozone-induced foliar injury on tall milkweed (Asclepias exaltata) in Great Smoky Mountains National Park. AB - The goals of this study were to document the development of ozone-induced foliar injury, on a leaf-by-leaf basis, and to develop ozone exposure relationships for leaf cohorts and individual tall milkweeds (Asclepias exaltata L.) in Great Smoky Mountains National Park. Plants were classified as either ozone-sensitive or insensitive based on the amount of foliar injury. Sensitive plants developed injury earlier in the season and to a greater extent than insensitive plants. Older leaf cohorts were more likely to belong to high injury classes by the end of each of the two growing seasons. In addition, leaf loss was more likely for older cohorts (2000) and lower leaf positions (2001) than younger cohorts and upper leaves, respectively. Most leaves abscised without prior ozone-like stippling or chlorosis. Failure to take this into account can result in underestimation of the effects of ozone on these plants. PMID- 16213639 TI - Characterization of polycyclic aromatic hydrocarbons in urban stormwater runoff flowing into the tidal Anacostia River, Washington, DC, USA. AB - To investigate the sources, fate, and transport dynamics of PAHs (polycyclic aromatic hydrocarbons) in stormwater runoff that is a leading source of pollution in urban watersheds, storm and base flow samples were collected in six branches along the lower Anacostia River. PAHs in storm flow (1510-12,500 ng/L) were significantly enriched in the particle phase, which accounted for 68-97% of the total PAHs. It suggests that reducing particles in stormwater using post treatment system would decrease PAHs considerably. The solid-water distribution coefficients (KD) of PAHs in the storm flow samples were up to 340 times higher than predicted values. A greater portion of high molecular weight PAHs and their distribution patterns indicate higher contribution of automobile originated pyrogenic PAHs. Total suspended solids in storm flow had a positive relationship with flow rates and exceeded benchmark level for the protection of aquatic biota in some samples. PMID- 16213640 TI - Response of drinking-water reservoir ecosystems to decreased acidic atmospheric deposition in SE Germany: trends of chemical reversal. AB - This study evaluates chemical trends of seven acidified reservoirs and 22 tributaries in the Erzgebirge from 1993 to 2003. About 85% of these waters showed significantly (p < 0.05) declining concentrations of protons (-69%), nitrate ( 41%), sulfate (-27%), and reactive aluminum (-50% on average). This reversal is attributed to the intense reduction of industrial SO2 and NOx emissions from formerly high levels, which declined by 99% and 82% in the German-Czech border region between 1993 and 1999. The deposition rates of protons and sulfur decreased by 70-90%. Since 1993, the dry deposition of total inorganic nitrogen diminished to a minor degree, but the wet deposition remained unchanged. The surface waters reflect a substantial decrease in Al exchange processes, a release of sulfur previously stored in soils, and an uptake of nitrate by forest vegetation. The latter effect may be supported by soil protection liming which contributed to the chemical reversal in almost 20% of the study waters. PMID- 16213641 TI - High average daily intake of PCDD/Fs and serum levels in residents living near a deserted factory producing pentachlorophenol (PCP) in Taiwan: influence of contaminated fish consumption. AB - An abandoned pentachlorophenol plant and nearby area in southern Taiwan was heavily contaminated by dioxins, impurities formed in the PCP production process. The investigation showed that the average serum PCDD/Fs of residents living nearby area (62.5 pg WHO-TEQ/g lipid) was higher than those living in the non polluted area (22.5 and 18.2 pg WHO-TEQ/g lipid) (P<0.05). In biota samples, average PCDD/F of milkfish in sea reservoir (28.3 pg WHO-TEQ/g) was higher than those in the nearby fish farm (0.15 pg WHO-TEQ/g), and Tilapia and shrimp showed the similar trend. The average daily PCDD/Fs intake of 38% participants was higher than 4 pg WHO-TEQ/kg/day suggested by the world health organization. Serum PCDD/F was positively associated with average daily intake (ADI) after adjustment for age, sex, BMI, and smoking status. In addition, a prospective cohort study is suggested to determine the long-term health effects on the people living near factory. PMID- 16213642 TI - "...it's all the same no matter how much fruit or vegetables or fresh air we get": traveller women's perceptions of illness causation and health inequalities. AB - This paper explores the perceptions of illness causation and health inequalities of Travellers, an ethnic minority group who experience considerable social and health disadvantages in Ireland. In order to allow for subjective meanings to emerge, a qualitative methodology with purposive sampling was employed. Participants in the study were invited to respond to a vignette in a focus group setting. Forty-one Traveller women were recruited to the focus groups through community projects or adult education initiatives. The study not only illustrates the complexity of lay perceptions of ill-health and health inequalities, but raises important questions about the prevalence of depression and of domestic violence in the Travelling community. These Traveller women were very willing to discuss the structural factors that contributed to their health status, attributing ill-health to social and environmental factors, such as accommodation, hardship and discrimination. Further, they broadly rejected behavioural explanations of the heart disease described in the vignette. Traveller women's understandings of health and the factors that determine it are deeply embedded in the social context of their lives and their ethnic identity. These findings are discussed in the context of social identity and ethnicity, and contribute to theoretical debates about the role of that identity in recognising inequality. The study revealed that Traveller women see many shortcomings in health service provision. They need service provision to be culturally sensitive and responsive to their needs. PMID- 16213643 TI - Turning the medical gaze in upon itself: root cause analysis and the investigation of clinical error. AB - In this paper, we discuss how a technique borrowed from defense and manufacturing is being deployed in hospitals across the industrialized world to investigate clinical errors. We open with a discussion of the levers used by policy makers to mandate that clinicians not just report errors, but also gather to investigate those errors using root cause analysis (RCA). We focus on the tensions created for clinicians as they are expected to formulate 'systems solutions' that go beyond blame. In addressing these matters, we present a discourse analysis of data derived during an evaluation of the NSW Health Safety Improvement Program. Data include transcripts of RCA meetings which were recorded in a local metropolitan teaching hospital. From this analysis we move back to the argument that RCA involves clinicians in 'immaterial labour', or the production of communication and information, and that this new labour realizes two important developments. First, because RCA is anchored in the principle of health care practitioners not just scrutinizing each other, but scrutinizing each others'errors, RCA is a challenging task. Second, thanks to turning the clinical gaze in on the clinical observer, RCA engenders a new level of reflexivity of clinical self and of clinical practice. We conclude with asking whether this reflexivity will lock the clinical gaze into a micro-sociology of error, or whether it will enable this gaze to influence matters superordinate to the specifics of practice and the design of clinical treatments; that is, the over arching governance and structuring of hospital care. PMID- 16213645 TI - Health care practices, professions and perspectives: a case study in intensive care. AB - This paper makes the case for bringing empirical analysis to the heart of conceptual work about health care practice. Drawing on ethnographic observations in intensive care units in the UK, it identifies and analyses a mismatch between the practice of a particular health care specialty and its associated professional and academic discourse. As a result, two claims in academic nursing discourse are criticised: first, that nursing practice is (or should be) focused on individual patient care; and second, that nursing is (or should be) radically distinct from medicine. By raising the analytical profile of situated practice, health care workers (in particular, but not exclusively, nurses) are shown to undertake important caring work with patients' relatives. Their work includes caring not only for an individual patient's social self, but also for patients'social contexts. The paper also argues that theoretical emphases on nursing's unique perspective and on differences between medicine and nursing are exaggerated in clinical practice, for example there are many similarities between what nurses and doctors actually do. Reasons for the persistence of these claims in academic nursing discourse are put forward-nursing seems to be quite unusual in needing an explicit theory of practice, and the paper speculates on why this is the case. The general lesson of the paper is that analytical evidence about the context and content of practice needs to be afforded a more fundamental role in the development of theories about practice-based disciplines. PMID- 16213644 TI - Socioeconomic status and health: do gradients differ within childhood and adolescence? AB - Socioeconomic status (SES) gradients may not be static across the lifespan, but instead may vary in strength across different life stages. This study examined the periods in childhood when SES and health relationships emerge and are strongest among US children. Data came from the National Health Interview Survey, 1994, a cross sectional, nationally representative sample of 33,911 US children ages 0-18. Parents were asked about family SES and child health status. Global health measures included overall ratings of child health, activity and school limitations. Acute conditions included childhood injuries and respiratory illnesses. For all global child health measures, lower family SES was associated with poorer child health in a gradient fashion (P < .001); these differences did not vary across age. For specific conditions, interaction effects of SES with age were found (P < .05). Interaction effects revealed that for injury and acute respiratory illness, expected SES gradients (lower SES with poorer outcomes) were evident during adolescence. In contrast, respiratory illness had a reverse SES gradient in early childhood. In sum, for global child health measures, associations of lower SES with poorer health throughout childhood suggest that factors that do not change with age (e.g., health care quality) may best explain overall health status. However, for acute conditions, the relationship between low SES and poor child health appears most consistently during adolescence. This suggests that normal development-related changes during adolescence, such as increasing peer group affiliation, may help explain these gradients. These patterns are important to understand for optimally timing interventions to reduce SES disparities in US children's health. PMID- 16213646 TI - Acute, subchronic and chronic safety studies with genistein in rats. AB - Genistein is a phytoestrogen that occurs naturally in the diet, especially in soy based foods. There is wide spread interest in phytoestrogens as chemopreventive agents for a variety of diseases and cancers based on epidemiologic evidence. Although soy, and its constituents such as genistein, have been consumed at high levels in several Asian populations without apparent adverse effects, concern has been raised about potential adverse effects due to the estrogenic and other activities. Safety studies with genistein were conducted in the Wistar rat including two acute studies, two subchronic (4 weeks and 13 weeks) and a chronic 52-week dietary admix study. In the acute studies, genistein had a low order of toxicity. In the three repeated dose safety studies at doses up to 500 mg/kg/day, genistein was well tolerated. In all of the studies, decreased food consumption and body weight gain were observed at 500 mg/kg/day. The main hematological findings were decreased red blood cell parameters at 500 mg/kg/day with a compensatory increase in reticulocytes. For clinical chemistry, with the exception of a slight increase in gamma glutamyl transferase in male and female rats at the high dose, there were a number of other minor changes considered not toxicologically significant. At necropsy, there were relatively few macroscopic changes; in the 52-week study, dilation of the uterus with fluid at the high dose and cysts of the ovaries in treated animals were observed. Organ weight changes in male rats at the high dose of 500 mg/kg/day included increased kidney, spleen, adrenal and testes weights and for females included, increased liver, kidney, spleen, ovary and uterus weights. After 4 and 13 weeks of treatment with genistein, there were no treatment related histopathologic findings. After 26 and 52 weeks of treatment, histological changes were seen in the female reproductive organs (ovaries and uterus), and in males (epididymides and prostate), and bone, kidneys, heart, liver and spleen in both sexes. After 52 weeks of treatment of males, vacuolation of the epididymal epithelium at 500 mg/kg/day and inflammation of the prostate were recorded at a higher incidence at 50 and 500 mg/kg/day. In females, cytological changes in the uterus, squamous metaplasia at 50 and 500 mg/kg/day and hyperplasia at 500 mg/kg/day were observed. Furthermore, hydrometra of the uterus and findings in the vagina consisting of anestric or diestrus vaginal mucosa with vaginal mucification, hyperplastic epithelium and multifocal cystic degeneration were noted at 500 mg/kg/day. Atrophy of the ovaries increased in severity in animals at 50 and 500 mg/kg/day. Osteopetrosis (hyperostosis) was observed in male and female rats at 50 and 500 mg/kg/day along with a compensatory increase in extramedullary hemopoiesis in the spleen; females were more affected than males. Hepatocellular hypertrophy and minimal bile duct proliferation were recorded at a higher incidence in animals at 500 mg/kg/day. It is concluded that almost all of the treatment related findings in these studies are related to the estrogenic properties of genistein as a phytoestrogen and would be expected to occur with a compound with estrogenic activity. The hormonally related changes were considered to be functional in nature and thus not adverse effects. Most of the findings in these studies were limited to the high dose of 500 mg/kg/day and were reversible. The few findings observed at 50 mg/kg/day were relatively minor and in view of the functional (hormonally mediated) nature of the effects, were considered not adverse effects. The increased incidence of minimal bile duct proliferation and slightly increased gamma glutamyl transferase are indicative of a mild hepatic effect at the high dose of 500 mg/kg/day. The no observed adverse effect level (NOAEL) of genistein is considered to be 50 mg/kg/day based on the presence of mild hepatic effects at the high dose of 500 mg/kg/day. The no observed effect level (NOEL) is considered to be 5 mg/kg/day based on the hormonally induced functional changes at higher doses. PMID- 16213647 TI - Low level lindane exposure alters extinction of conditioned fear in rats. AB - Gamma-hexachlorocyclohexane (lindane) is a pesticide with the potential to produce long-term effects on fear or anxiety due to its targeting of the GABA(A) receptor in the brain. Multiple chemical sensitivity (MCS) is a human condition that has been attributed to repeated chemical exposures, with pesticides heavily implicated in the initiation of MCS. The symptoms in MCS patients are wide ranging but prominent among these in a subset of patients is increased evoked panic responses. Drawing a parallel between these responses in MCS patients and a panic model in rats, these studies explored a potential animal model for MCS. The effects of repeated lindane exposure on conditioned fear behavior was examined in adult male Sprague-Dawley rats. Animals were administered vehicle or lindane (intraperitoneally) for either 3 days/week (1, 2 or 5mg) or 5 days/week (2mg) over 2 weeks, and 18 days later were examined for anxiety levels on an elevated plus-maze. One day later, animals were trained for fear conditioning to an odor conditioned stimulus (CS). Freezing behavior was measured 1 day later in the context where pairing occurred, and then for a total of 6 days in a different environment in which either no CS or the CS was presented. After a second 18-day period of no treatment, rats were again tested for their freezing response to the CS for 2 days. Lindane pretreatment did not alter elevated plus-maze performance, nor did it alter contextual freezing behavior. However, pretreatment with lindane decreased the extinction of fear conditioning to the CS such that freezing behavior in controls was significantly lower than in lindane-pretreated rats, and this effect persisted during testing 18 days later. The results indicate that repeated low-level lindane exposure may produce long-lasting changes in anxiety related neural circuitry. This suggests that odor-triggered symptoms associated with an aversive event may persist in MCS patients because of the ability of some chemicals to alter fear or anxiety circuitry in the brain. PMID- 16213649 TI - Self-association and DNA binding properties of the human topoisomerase IIA alpha2HTH module. AB - The eukaryotic topoisomerase II is an ubiquitous nuclear enzyme involved in vital cellular functions. It is also the target for some of the most active anticancer drugs. In the various crystal structures of yeast topoisomerase II, the 701-748 segment homologous to the human topoisomerase II alpha 724-771 segment folds into a compact alpha(2)beta(1)alpha(3)talpha(4) conformation, hereafter termed alpha(2)HTH module (helix turn helix (HTH), alpha(3)talpha(4)). The crystal structure of gyrase A has suggested a model wherein HTH is involved in both the enzyme dimerization and the binding to DNA. These two properties were investigated in solution, using the recombinant alpha(2)HTH module of human topoisomerase II alpha and its synthetic components HTH, alpha(4), alpha(3) and turn. The homology-based structure model of human alpha(2)HTH superposed that of yeast in the crystal structure with a rmsd of 1.03 A. Circular dichroism spectra showed that the helical content of human alpha(2)HTH in solution is similar to that of its counterpart within yeast topoisomerase II in the solid state. The chemical cross-linking data indicated that alpha(2)HTH self-associated into dimers while gel mobility shift assays and anisotropy fluorescence titrations demonstrated that alpha(2)HTH, HTH and alpha(4), but not alpha(3), bind efficiently to DNA (dissociation constants of 3.10(-7) M for alpha(2)HTH and alpha(4), of 3.10(-6) M for HTH and of only 1.10(-5) M for alpha(3)). Correlatively, alpha(2)HTH, alpha(4) and HTH, but not alpha(3), were able to inhibit topoisomerase II in DNA relaxation assays, stipulating that alpha(4) is the DNA recognition helix. All suggests that the alpha(2)HTH module once separated from the whole protein conserves a compact conformation, integral to specific dimerization and DNA recognition. The module may thus be used for the search of drugs efficient in hindering topoisomerase II dimerization or binding to DNA. PMID- 16213648 TI - Characterization of anti-apoptotic action of TCDD as a defensive cellular stress response reaction against the cell damaging action of ultra-violet irradiation in an immortalized normal human mammary epithelial cell line, MCF10A. AB - It was originally shown by Woerner and Schrenk [Woerner, W., Schrenk, D., 1998. 2,3,7,8-Tetrachlorodibenzo-p-dioxin suppresses apoptosis and leads to hyperphosphorylation of p53 in rat hepatocytes. Environ. Toxicol. Pharmacol. 6, 239-247] that TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) acts as an antagonist against the action of UV-irradiation to induce apoptosis in rat primary hepatocytes. Since prevention of apoptosis has been shown to promote carcinogenesis, we have decided to investigate this phenomenon in a human mammary gland epithelial cell line, MCF10A. We found that, in this cell line, TCDD can antagonize apoptosis that was induced by a variety of treatments, such as UV- and gamma-irradiation, growth factor starvation and trypsinization, or by the addition of H(2)O(2), TGFbeta, and staurosporine. Furthermore, other agents that are known to elicit defensive cellular responses, such as LPS, Fe(3+), nitric oxide and hypoxia could also antagonize UV induced apoptosis just as in the case of TCDD. In addition, we found that, in this cell line, such anti-apoptotic action of TCDD resembles that of exogenously added EGF or TGF alpha. To study the basic mechanism of such an action of TCDD, we tested a variety of diagnostic agents to reverse the effect of TCDD. Antagonists of TCDD which were found to be effective in this way were (a) inhibitors of c-Src kinase, such as PP-2 and CGP77675, (b) those known to block the action of TGF alpha, such as anti-TGF alpha antibody, and alpha(1)-antitrypsin, (c) PD98059, a specific inhibitor of ERK activation, but not SB202190 (an inhibitor of p38 MAPK activation) or SP600125 (a JNK inhibitor) and (d) Ah receptor antagonists, alpha-naphthoflavone and 1, 10-phenanthroline. These results support the notion that TCDD acts as an anti-apoptotic agent by mimicking the action of EGF through activation of the c Src/ERK signaling pathway. PMID- 16213650 TI - Identification and biochemical characterization of Rap2C, a new member of the Rap family of small GTP-binding proteins. AB - The Rap family of small GTP-binding proteins is composed by four different members: Rap1A, Rap1B, Rap2A and Rap2B. In this work we report the identification and characterization of a fifth member of this family of small GTPases. This new protein is highly homologous to Rap2A and Rap2B, binds labeled GTP on nitrocellulose, and is recognized by a specific anti-Rap2 antibody, but not by an anti-Rap1 antibody. The protein has thus been named Rap2C. Binding of GTP to recombinant purified Rap2C was Mg(2+)-dependent. However, accurate comparison of the kinetics of nucleotide binding and release revealed that Rap2C bound GTP less efficiently and possessed slower rate of GDP release compared to the highly homologous Rap2B. Moreover, in the presence of Mg(2+), the relative affinity of Rap2C for GTP was only about twofold higher than that for GDP, while, under the same conditions, Rap2B was able to bind GTP with about sevenfold higher affinity than GDP. When expressed in eukaryotic cells, Rap2C localized at the plasma membrane, as dictated by the presence of a CAAX motif at the C-terminus. We found that Rap2C represented the predominant Rap2 protein expressed in circulating mononuclear leukocytes, but was not present in platelets. Importantly, Rap2C was found to be expressed in human megakaryocytes, suggesting that the protein may be down-regulated during platelets generation. This work demonstrates that Rap2C is a new member of the Rap2 subfamily of proteins, able to bind guanine nucleotides with peculiar properties, and differently expressed by various hematopoietic subsets. This new protein may therefore contribute to the still poorly clarified cellular events regulated by this subfamily of GTP-binding proteins. PMID- 16213651 TI - Interplay between the cis-prenyltransferases and polyprenol reductase in the yeast Saccharomyces cerevisiae. AB - Dolichol formation is examined in three Saccharomyces cerevisiae strains with mutations in the ERG20 gene encoding farnesyl diphosphate synthase (mevalonic acid pathway) and/or the ERG9 gene encoding squalene synthase (sterol synthesis pathway) differing in the amount and chain length of the polyisoprenoids synthesized. Our results suggest that the activities of two yeast cis prenyltransferases Rer2p and Srt1p and polyprenol reductase are not co-regulated and that reductase may be the rate-limiting enzyme in dolichol synthesis if the amount of polyisoprenoids synthesized exceeds a certain level. We demonstrate that reductase preferentially acts on typical polyprenols with 13-18 isoprene residues but can reduce much longer polyprenols with even 32 isoprene residues. PMID- 16213652 TI - Modeling and simulation in signal transduction pathways: a systems biology approach. AB - Modeling, the heart of systems biology, of complex processes (example: signal transduction) is a wide scientific discipline where many approaches from different areas are confronted with the aim of better understanding, identifying and modeling of complex data coming from various sources. The purpose of this paper is to introduce the basic steps of systems biology view towards signaling pathways, which mainly deals with the computational tools. The paper emphasizes the modeling and simulation approach in the signal transduction pathways using the topologies of the biochemical reactions with an overview of the different types of software platforms. Finally, we demonstrated the epidermal growth factor receptor signaling pathway model as an example to study the growth factor mediated signaling system with biological experiments. This paper will enables new comers to underline the strengths of the computational approaches towards signal transduction, as well as to highlight the systems biology research directions. PMID- 16213653 TI - The insular lobe and brain plasticity: Lessons from tumor surgery. AB - OBJECTIVES: Despite recent literature supporting the likely role of the insula in many functions, the actual participation of this multimodal lobe in the brain functioning remains unclear, i.e. has the insula an essential or compensable role? PATIENTS AND METHODS: We surgically resected an insular low-grade glioma, using intraoperative electrical stimulation, in 42 patients who experienced seizures, but who presented no or only a slight neurological deficit. Surgery was performed under local anesthesia in patients with a lesion in the dominant hemisphere. The resection was systematically stopped according to cortico subcortical functional boundaries. RESULTS: Intraoperative electrical mapping induced language disturbances, pain and vertigo, but no other side effects were observed. Post-operatively, the patients experienced a transient hemiparesis in 21 cases, language disorders in 10 cases, an athymhormic syndrome in 7 cases, a Foix-Chavany-Marie syndrome in 3 cases, and micturition disturbances in one case. Despite this immediate post-surgical worsening, all the patients recovered their preoperative neurological status within 3 months, except in three cases due to a deep stroke. CONCLUSION: These results show that the insula, a complex associative multimodal structure poorly studied until now, can be functionally compensated. Such a plastic potential may have important fundamental and clinical implications, in particular in the field of oncological neurosurgery. PMID- 16213654 TI - Genetically induced retinal degeneration leads to changes in metabotropic glutamate receptor expression. AB - In the retina, neurotransmission from photoreceptors to ON-cone and rod bipolar cells is sign reversing and mediated by the metabotropic glutamate receptor mGluR6, which converts the light-evoked hyperpolarization of the photoreceptors into depolarization of ON bipolar cells. The Royal College of Surgeons (RCS) rat retina undergoes progressive photoreceptor loss due to a genetic defect in the pigment epithelium cells. The consequences of photoreceptor loss and the concomitant loss of glutamatergic input to second-order retinal neurons on the expression of the metabotropic glutamate receptor was investigated in the RCS rat retina from early stages of photoreceptor degeneration (P17) up to several months after complete rod and cone degeneration (P120). The expression of the gene encoding mGluR6 was studied by in situ hybridization in the retina, using an [(35)S]dATP-labeled oligonucleotide probe. In congenic control and RCS retina, we found mRNA expression of mGluR6 receptor only in the outer half of the inner nuclear layer (INL) on emulsion-coated retinal sections. Quantitative analysis of the hybridization signal obtained from the autoradiographic films revealed decreased expression levels of the mGluR6 mRNA at early stages of photoreceptor degeneration (P17). On the contrary, increased expression levels were observed at late stages of degeneration (P60 and P120) in RCS compared to congenic control retina. In conclusion, our data demonstrate that the metabotropic glutamate receptor-6 mRNA levels are altered in the young and adult RCS rat retina and suggest that the genetically induced degeneration of photoreceptors affects the expression of this receptor by the INL retinal neurons. PMID- 16213655 TI - Temporal aspects of the visuotactile congruency effect. AB - We report an experiment designed to investigate the temporal dynamics of the visuotactile crossmodal congruency effect. Vibrotactile targets were presented randomly to the index finger (top side of a hand-held cube) or thumb (bottom side) of either hand while visual distractors were presented randomly from one of the same four possible locations. The stimulus onset asynchrony (SOA) between the vibrotactile target and the visual distractor was varied on a trial-by-trial basis. Participants made speeded discrimination responses regarding the elevation of the vibrotactile targets (i.e., upper versus lower) while trying to ignore the visual distractors. The largest crossmodal congruency effects (defined as the difference in performance between incongruent and congruent elevation distractor trials) were obtained when the visual distractor preceded the vibrotactile target by 50-100 ms, although significant effects were also reported when the distractor followed the target by as much as 100 ms. These results are discussed in terms of the conjoint influence of response competition, crossmodal perceptual interactions (i.e., the ventriloquism effect), and exogenous spatial attention on the crossmodal congruency effect. The distinct temporal signatures of each of these effects are also highlighted. PMID- 16213656 TI - Audiovisual synchrony perception for speech and music assessed using a temporal order judgment task. AB - This study investigated people's sensitivity to audiovisual asynchrony in briefly presented speech and musical videos. A series of speech (letters and syllables) and guitar and piano music (single and double notes) video clips were presented randomly at a range of stimulus onset asynchronies (SOAs) using the method of constant stimuli. Participants made unspeeded temporal order judgments (TOJs) regarding which stream (auditory or visual) appeared to have been presented first. The accuracy of participants' TOJ performance (measured in terms of the just noticeable difference; JND) was significantly better for the speech than for either the guitar or piano music video clips, suggesting that people are more sensitive to asynchrony for speech than for music stimuli. The visual stream had to lead the auditory stream for the point of subjective simultaneity (PSS) to be achieved in the piano music clips while auditory leads were typically required for the guitar music clips. The PSS values obtained for the speech stimuli varied substantially as a function of the particular speech sound presented. These results provide the first empirical evidence regarding people's sensitivity to audiovisual asynchrony for musical stimuli. Our results also demonstrate that people's sensitivity to asynchrony in speech stimuli is better than has been suggested on the basis of previous research using continuous speech streams as stimuli. PMID- 16213657 TI - Assessing functioning of the prefrontal cortical subregions with auditory evoked potentials in sleep-wake cycle. AB - Our previously observations showed that the amplitude of cortical evoked potentials to irrelevant auditory stimulus (probe) recorded from several different cerebral areas was differentially modulated by brain states. At present study, we simultaneously recorded auditory evoked potentials (AEPs) from the dorsolateral prefrontal cortex (DLPFC) and the ventromedial prefrontal cortex (VMPFC) in the freely moving rhesus monkey to investigate state-dependent changes of the AEPs in the two subregions of prefrontal cortex. AEPs obtained during passive wakefulness (PW), active wakefulness (AW), slow wave sleep (SWS) and rapid-eye-movement sleep (REM) were compared. Results showed that AEPs from two subregions of prefrontal cortex were modulated by brain states. Moreover, a significantly greater increase of the peak-to-peak amplitude (PPA) of N1-P1 complexes appears in the DLPFC during PW compared to that during AW. During REM, the PPA of N1-P1 complexes presents a contrary change in the two subregions with significant difference: a significant increase in the DLPFC and a slight decrease in the VMPFC compared to that during AW. These results indicate that the modulation of brain states on AEPs from two subregions of the prefrontal cortex investigated is also not uniform, which suggests that different subregions of the prefrontal cortex have differential functional contributions during sleep-wake cycle. PMID- 16213658 TI - Central serotonin depletion affects rat brain areas differently: a qualitative and quantitative comparison between different treatment schemes. AB - Depletion of rat brain serotonin (5-hydroxytryptamin, 5-HT) has been widely used to study effects of serotonin and its interaction with other transmitter systems. Various treatment regimes for serotonin depletion have been applied, but the efficacy of these seems to vary considerably. So far, no studies have systematically examined and compared different approaches. The present work combines quantitative and qualitative measurements and compares six different treatment schemes for 5-HT depletion. Treatment outcome was evaluated by HPLC measurements of 5-HT and 5-HIAA concentrations, and by 5-HT and tyrosine hydroxylase immunocytochemistry. The schemes included repeated administration of fenfluramine (FEN) and/or p-chlorophenylalanine (pCPA). The most efficient treatment for rat brain 5-HT depletion was the combined treatment with one daily pCPA (200 mg/kg) injection for 3 days followed by one injection of d,l-FEN (20 mg/kg) on the fourth day, causing a 94.9% brain 5-HT depletion. Immunostaining revealed a distinct brain distribution of the remaining 5-HT, with an almost complete depletion of 5-HT in the cerebral cortex, hippocampus and thalamus, while a substantial amount of 5-HT still was left in the raphe nuclei, the medial forebrain bundle, and the medial eminence. FEN or pCPA treatment alone caused from 68.2 to 94.0% decrease in 5-HT levels. While the pattern of 5-HT depletion using pCPA alone was comparable to the one seen with the combined treatment, the 5-HT depletion using FEN alone showed a different pattern with 5-HT distributed in several brain regions. PMID- 16213659 TI - Algorithm for neuropathic pain treatment: an evidence based proposal. AB - New studies of the treatment of neuropathic pain have increased the need for an updated review of randomized, double-blind, placebo-controlled trials to support an evidence based algorithm to treat neuropathic pain conditions. Available studies were identified using a MEDLINE and EMBASE search. One hundred and five studies were included. Numbers needed to treat (NNT) and numbers needed to harm (NNH) were used to compare efficacy and safety of the treatments in different neuropathic pain syndromes. The quality of each trial was assessed. Tricyclic antidepressants and the anticonvulsants gabapentin and pregabalin were the most frequently studied drug classes. In peripheral neuropathic pain, the lowest NNT was for tricyclic antidepressants, followed by opioids and the anticonvulsants gabapentin and pregabalin. For central neuropathic pain there is limited data. NNT and NNH are currently the best way to assess relative efficacy and safety, but the need for dichotomous data, which may have to be estimated retrospectively for old trials, and the methodological complexity of pooling data from small cross-over and large parallel group trials, remain as limitations. PMID- 16213661 TI - Immune response to Leishmania infantum in healthy horses in Spain. AB - Leishmania infantum infection has recently been described in horses in Europe. We report the results of a study on the immune response to L. infantum in horses living in an area endemic for leishmaniosis in NE Spain. Two ELISAs using protein A and anti-horse IgG conjugates were adapted to measure specific antibodies to L. infantum in horse sera. A lymphocyte proliferation assay (LPA) of peripheral blood mononuclear cells to L. infantum antigen was also performed to detect specific cellular immune response to Leishmania. Anti-L. infantum antibodies were detected in the serum of 16 of the horses studied (n=112) using the protein A assay but not in the assay using the anti-horse IgG conjugate. Specific lymphocyte proliferation was observed in 20 out of 55 horses. This study shows that horses in the area studied mount specific immune responses to L. infantum, and must therefore be considered among the species exposed to the parasite in this region. The infrequency of leishmaniosis in horses suggests that the immune response in this species is effective in controlling the infection. PMID- 16213663 TI - Metabolic changes in elderly patients with major depression: evidence for increased accumulation of visceral fat at follow-up. AB - The accumulation of visceral fat is promoted by a specific endocrine syndrome, which is similarly found in major depression. The aim of this study was to investigate whether visceral fat depots increase in depressed patients during a follow-up period explaining the increased risk for cardiovascular disorders. Intraabdominal fat was measured in 29 depressed patients and 17 controls by computer tomography at the level of lumbar vertebra 4. In patients fat measurements were done initially during a major depressive episode and again after a follow-up period of 14 months; in controls the mean time interval between measurements was 28 months. In both groups, saliva was taken at 800 h over a period of seven days prior to each CT for the estimation of free cortisol. In patients only, an oral glucose tolerance test was also carried out. Compared to controls hyper- and normocortisolemic depressed patients showed a larger accumulation of visceral fat mass over time (hypercort.:132.0 +/- 45 vs. 144.7 +/ 47 cm(2), p = 0.07; normocort.: 115.5 +/- 53 vs. 135.0 +/- 51 cm(2), p = 0.002; controls: 130.1 +/- 66 vs. 137.3 +/- 76 cm(2), p = 0.4), despite similar weight gain (hypercort.: 2.1 +/- 5 kg, normocort.: 1.7 +/- 5 kg and controls: 2.3 +/- 4 kg). Further, normocortisolemic patients showed a trend for an higher percentile increase in visceral fat accumulation than controls (23.9 +/- 27 vs. 5.8 +/- 28%, p = 0.07). At follow-up, free cortisol concentrations were still above normal in patients who had been hypercortisolemic at first assessment (35.0 +/- 8 vs. 28.8 +/- 18 nmol/l, p = 0.1). Fasting and 2 h glucose concentrations were higher in hypercortisolemic compared to normocortisolemic patients at the index examination (6.2 +/- 1.1 vs. 5.0 +/- 0.05 mmol/l, p = 0.02; 11.5 +/- 2.7 vs. 7.8 +/- 1.9 mmol/l, p = 0.01). The larger proportion of visceral fat accumulation in patients may constitute a link for explaining the increased cardiovascular mortality in patients suffering from major depression. PMID- 16213664 TI - Is sleep's 'supreme mystery' unraveling? An evolutionary analysis of sleep encounters no mystery; nor does life's earliest sleep, recently discovered in jellyfish. AB - Biotelemetry has revealed daily 15-h behavioral sleep periods in a cubomedusan jellyfish, Chironex fleckeri. Its sleep is expected to be phylogenetically most primitive, since jellyfish possess only two germ layers. They belong to the phylum Cnidaria, the 'simplest' multicellular organisms with an organized nervous system. Cubomedusae have a complex visual system with 24 eyes of four different types, each type specialized for a different task. Input to these eyes during visually guided fast-swimming predation requires enormous amounts of neural processing, possibly nearly saturating the capacity of their comparatively simple nervous system. These heavy neural demands may account for the need for fifteen hours of sleep. C. fleckeri is the only animal known for which sleep may be either present or absent, dependent on lifestyle. Limited knowledge of behavior of some other cubomedusae suggests that they also possess this faculty. The finding of sleep in C. fleckeri supports current proposals of sleep's origin and basic function. Evolutionary analyses link sleep to a conflict produced by excessive processing demands on multifunctional neural circuitry for detailed focal vision by complex lensed eyes. The conflict arises between the enormous demands of complex visual analysis and needs for split-second control of actions, on the one hand, and non-urgent processing of memories of ongoing and stored events, on the other. Conflict is resolved by deferring the non-urgent processing to periods of sleep. Without sleep, selection would favor the evolution of circuitry 'dedicated' to single or but few tasks, with corresponding lesser efficiency. Had complex lensed eyes of medusae originated as a consequence of selection for increased mating success of males pursuing females, it could have occurred before the evolution of fast-swimming bilateral (three-germ-layered) prey. But if it was a consequence of selection for increased prey-hunting success, the origin of such eyes probably awaited the coexistence of bilateral prey. PMID- 16213662 TI - Flavonoids purified from Rhus verniciflua Stokes actively inhibit cell growth and induce apoptosis in human osteosarcoma cells. AB - Many studies have suggested that dietary flavonoids are anticancer agents that induce the apoptosis of cancer cells. However, the effects of flavonoids on the induction of apoptosis in osteosarcoma cells are unclear. Previously, a flavonoid fraction, consisting mainly of protocatechuic acid, fustin, fisetin, sulfuretin, and butein, herein named RCMF (the RVS chloroform-methanol fraction), was prepared from a crude acetone extract of Rhus verniciflua Stokes (RVS). This study evaluated the effects of RCMF on the proliferation and apoptosis using human osteosarcoma (HOS) cells. The mechanism of growth inhibition of the HOS cells by the flavonoid fraction, RCMF, was also assessed. The results demonstrated that RCMF exhibited sensitive growth inhibition and induced apoptosis in HOS cells. PARP cleavage was closely associated with the RCMF induced apoptosis of the HOS cells. Furthermore, the activation of caspase 8 and Bax, the inhibition of Bcl-2 expression, and the release of cytochrome c are believed to be involved in the RCMF-mediated apoptosis. Collectively, these findings suggest that RCMF is an agent which may be capable of inducing sensitive growth inhibition and apoptosis in HOS cells. PMID- 16213665 TI - The future of selective serotonin reuptake inhibitors (SSRIs) in psychiatric treatment. AB - Selective serotonin reuptake inhibitors (SSRIs) are widely prescribed and widely regarded as a first-line treatment for depression. Yet, a growing body of evidence indicates that these agents are only moderately more effective than placebo in treating major depressive disorder. In recent years, it has been debated whether SSRIs offer any clinically meaningful advantage over placebos. As part of this debate, it has been argued that these agents are first-line treatments for some forms of depression but not necessarily for others. The present paper examines two hypotheses that are central to these issues. The first hypothesis is that SSRIs are more effective than placebo for some types of depression but not for others. The second is that SSRIs are more effective than psychotherapies for some types of depression than others. A review of the empirical literature reveals three main classifications of depression that are relevant to the first hypothesis: (a) more vs. less severe depression, (b) melancholic vs. non-melancholic depression, and (c) depression defined according to associated genetic factors (particularly the long vs. short allele of the serotonin transporter gene promoter). There is no strong or consistent support for (a) or (b). There is, however, emerging and consistent evidence for (c), and so the first hypothesis is tentatively supported, but only for (c). Most of the empirical evidence does not support the second hypothesis. Psychotherapies (cognitive-behavioral and interpersonal therapies) and SSRIs generally have equivalent efficacy, regardless of the severity of depression. The research literature also suggests a third hypothesis that remains to be evaluated: that SSRIs are more effective for treating anxiety disorders (and possibly other disorders) than they are for treating depression. If that hypothesis is supported by subsequent research, then the future of SSRIs may lie largely in the treatment of anxiety disorders, and in the management of particular subtypes of depression. PMID- 16213666 TI - No anti-ACTH autoantibody in serum of SARS patients. PMID- 16213667 TI - PSA kinetics after prostate brachytherapy: PSA bounce phenomenon and its implications for PSA doubling time. AB - PURPOSE: To analyze prostate-specific antigen (PSA) kinetics in patients treated with prostate brachytherapy (PI) with a minimum of 5 years of PSA follow-up. METHODS AND MATERIALS: The records of 162 patients treated with PI for localized prostate cancer with a minimum of 5 years of PSA follow-up were reviewed. A variety of pretreatment and posttreatment variables were examined. Patients were coded as having a PSA bounce if their PSA achieved a nadir, elevated at least 0.2 ng/mL greater than that nadir, and decreased to, or below, the initial nadir. Two definitions of biochemical failure (bF) or biochemical relapse-free survival (bRFS) were used: the classic American Society for Therapeutic Radiology and Oncology consensus definition of three consecutive rises (bF3) and the nadir plus 2 ng/mL definition (bFn+2). Associations between a PSA bounce and the various pre and posttreatment factors were assessed with logistic regression analysis, and the association between a PSA bounce and bF was examined with the log-rank test. The Mann-Whitney U test was applied to test for differences in the PSA doubling time (PSADT) and the time to a PSA rise between the PSA bounce patients and the bF patients. PSADT was calculated from the nadir to the time of the first PSA rise, because this point is known first in the clinical setting. RESULTS: The 5 year overall bRFS rate was 87% for the bF3 definition and 96% for the bFn+2 definition. A PSA bounce was experienced by 75 patients (46.3%). Patients who experienced a PSA bounce were less likely to have a bF, regardless of the bRFS definition used (bF3: p=0.0015; bFn+2: p=0.0040). Among the pre- and posttreatment factors, only younger age predicted for a PSA bounce on multivariate analysis (p=0.0018). The use of androgen deprivation had no effect on PSA bounce. No difference was found in the PSADT between patients who had a PSA bounce and those with bF. The median PSADT for those with a PSA bounce was 8.3 months vs. 10.3 months using the bF3 definition and 8.8 months using the bFn+2 definition. However, a significant difference was found in the time to the first rise in PSA after PI for patients with a PSA bounce vs. patients with bF. The median time to the first rise in PSA after nadir for those with a PSA bounce was 15.1 months vs. 30.0 months using the bF3 definition (p=0.001) and 22.3 months using the bFn+2 definition (p=0.013). CONCLUSION: Patients experiencing a PSA bounce are more likely to be younger and will have a better bRFS. The PSADT cannot differentiate a PSA bounce from bF. The time to the initial PSA rise after nadir is an excellent discriminator of bF from PSA bounce. The time of the PSA rise after nadir occurs far sooner for a PSA bounce than for bF. This factor should be considered when assessing a patient with a rising PSA level after PI before a patient is administered salvage therapy. PMID- 16213668 TI - Response regulator DegU of Listeria monocytogenes regulates the expression of flagella-specific genes. AB - An isogenic mutant of Listeria monocytogenes EGD with a deletion of the response regulator gene degU showed a lack of motility due to the absence of flagella. In the present study, we used two-dimensional gel electrophoresis, mass-spectrometry and microarray analyses to identify the listerial genes that depend on DegU for expression. We found that the two L. monocytogenes operons encoding flagella specific genes and the monocistronically transcribed flaA gene are positively regulated by DegU at 24 degrees C, but are not expressed at 37 degrees C. PMID- 16213669 TI - Relative contribution of target gene mutation and efflux to varying quinolone resistance in Irish Campylobacter isolates. AB - The contribution of target gene mutations and active efflux to varying levels of quinolone resistance in Irish Campylobacter isolates was studied. The Thr-86-Ile modification of GyrA did not correlate with the level of quinolone resistance. The efflux pump inhibitor Phe-Arg-beta-Naphthylamide (PAbetaN) had no effect on the MICs to ciprofloxacin. In contrast, a PAbetaN sensitive efflux system contributed to the low-level nalixidic acid resistance phenotype. The lack of effect of PAbetaN in high-level resistant nalidixic isolates may be attributable to mutations identified in the CmeB efflux pump of these isolates. PAbetaN may have limited diagnostic value in the assessment of the contribution of efflux pump activity to ciprofloxacin resistance in Campylobacter. PMID- 16213670 TI - Construction of the first shuttle vectors for gene cloning and homologous recombination in Mycoplasma agalactiae. AB - Mycoplasma agalactiae is a worldwide ruminant pathogen that causes significant economic losses by inflicting contagious agalactia in sheep and goats. The development of efficient control strategies requires a better understanding of the mycoplasma factors that promote successful infection. However, lack of genetic tools has been a major impediment in studying the pathogenic mechanisms of M. agalactiae. This study describes the identification and cloning of the M. agalactiae origin of replication (oriC) in order to construct the first shuttle vectors for targeted gene disruption, gene complementation and expression studies. Additionally, this report provides the first evidence of the occurrence of homologous recombination and the functionality of heterologous tetM determinant in this pathogen. PMID- 16213671 TI - Determination of coenzyme A levels in Pyrococcus furiosus and other Archaea: implications for a general role for coenzyme A in thermophiles. AB - Physiologically significant levels of intracellular coenzyme A were identified in Pyrococcus furiosus, Thermococcus litoralis, and Sulfolobus solfataricus, suggesting a role for CoA as an important low molecular mass thiol in the thermophilic Archaea. In P. furiosus, cells grown in the presence of sulfur showed significantly higher levels of oxidized CoA compared with those grown in the absence of S(0). T. litoralis showed strikingly similar CoA levels, although with low disulfide levels in both the presence and absence of S(0). S. solfataricus showed similarly high levels of CoA thiol, with correspondingly low levels of the CoA disulfide. These results are consistent with the identification of a coenzyme A disulfide reductase (CoADR) in P. furiosus and horikoshii as well as the presence of CoADR homologues in the genomes of S. solfataricus and T. kodakaraensis. PMID- 16213672 TI - Molecular cloning and functional analysis of (R)-3-hydroxyacyl-acyl carrier protein:coenzyme A transacylase from Pseudomonas mendocina LZ. AB - An inactive (R)-3-hydroxyacyl-acyl carrier protein:coenzyme A transacylase (PhaG(Pm)) was cloned from a newly isolated Proteobacteria Pseudomonas mendocina LZ. It is the first characterized native inactive PhaG protein. Sequence analysis indicated that there were only two sites where the amino acid sequence differed between this inactive protein and the functional PhaG(Pp) from P. putida. The differences were located at position 78 and in the region 109-113 in the amino acid sequence. Mutagenesis was carried out to investigate these two sites. A recombinant strain harboring a S78C PhaG(Pp) mutant accumulated polyhydroxyalkanoates (PHA) at 11.9% of the cellular dry weight, as compared to the 21.6% PHA produced by the recombinant harboring the wild-type PhaG(Pp). On the other hand, the changes in the amino acid region 109-113 of PhaG(Pp) to its corresponding region of PhaG(Pm) resulted in negligible PHA accumulation. This demonstrated that region 109-113 in PhaG is relatively important for transacylase activity, while position 78 just plays a supporting role for the enzyme. Furthermore, 3-D structural models of PhaG(Pp) and PhaG(Pm) developed by computational prediction revealed that the variation in amino acids at 109-113 leads to the destruction of the PhaG catalytic center, resulting in the loss of enzyme activity. PMID- 16213673 TI - ClpXP controls the expression of LEE genes in enterohaemorrhagic Escherichia coli. AB - Enterohaemorrhagic Escherichia coli (EHEC) contains a 36-kb pathogenicity island termed the locus of enterocyte effacement (LEE), which encodes a type III secretion system (TTSS) and virulence proteins. In this paper, we show that the O157:H7 Sakai clpPX mutant strongly impaired the secretion of virulence proteins by TTSS and repressed transcription from all the LEE promoters. The rpoS mutation in O157:H7 Sakai enhanced the transcription from all the LEE promoters and the secretion of virulence proteins, and it could partially suppress the defects of the clpPX mutation. These data indicate that the O157:H7 Sakai ClpXP protease is a positive regulator for LEE expression and that this regulation occurs by two pathways: the sigma(S)-dependent and -independent pathways. PMID- 16213674 TI - Candida dubliniensis: ten years on. AB - Candida dubliniensis was first described as a novel species in 1995. This organism is very closely related to the important human yeast pathogen, Candida albicans. However, despite the very close phylogenetic relationship between C. albicans and C. dubliniensis and the fact that they share a large number of phenotypic traits, epidemiological and virulence model data indicate that the former is a far more successful pathogen. In order to investigate the molecular basis of the lower virulence of C. dubliniensis recent comparative genomic hybridisation studies have revealed the absence and divergence of specific genes implicated in candidal virulence. Data from the C. dubliniensis genome sequencing project will allow a complete comparison between the genomes of the two species to be performed and thus enhance our understanding of candidal virulence and how virulence has evolved in Candida species. PMID- 16213675 TI - An outbreak of staphylococcal food poisoning caused by enterotoxin H in mashed potato made with raw milk. AB - Mashed potato made with raw bovine milk was suspected to have been the source of a food poisoning outbreak. Almost 8 x 10(8)Staphylococcus aureus CFU g(-1) were detected in the mashed potato. S. aureus was also found in bulk milk from the farm that had supplied milk for the mashed potato. Isolates from mashed potato and bulk milk were positive for the gene encoding staphylococcal enterotoxin H (seh), and the corresponding protein toxin, SEH, was detected by ELISA in the mashed potato. Production of SEH by S. aureus isolates from mashed potato (n = 4) and bulk milk (n = 4) was also demonstrated by ELISA. Sequencing of seh from one mashed potato isolate and one bulk milk isolate confirmed that the gene was a variant seh, and that the genes in both isolates were identical. Macrorestriction of chromosomal DNA with Sma1 followed by pulsed-field gel electrophoresis of seh positive S. aureus from mashed potato and bulk milk revealed indistinguishable banding patterns between isolates from both sources. It seems likely that raw bovine milk used in the preparation of mashed potato contained S. aureus that subsequently produced sufficient SEH in the mashed potato to cause food poisoning. PMID- 16213676 TI - Phage release from biofilm and planktonic Staphylococcus aureus cells. AB - The ability of pathogenic staphylococci to form biofilms facilitates colonization and the development of chronic infections. Therapy is hampered by the high tolerance of biofilms towards antibiotic treatment and the immune system. We found evidence that lysogenic Staphylococcus aureus cells in a biofilm and in planktonic cultures spontaneously release phages into their surroundings. Phages were detected over a much longer period in biofilm cultures than in planktonic supernatants because the latter were degraded by secreted proteases. Phage release in planktonic and biofilm cultures was artificially increased by adding mitomycin C. Two morphologically distinct phages in the S. aureus strain used in this work were observed by electron microscopy. We postulate that phage-release is a frequent event in biofilms. The resulting lysis of cells in a biofilm might promote the persistence and survival of the remaining cells, as they gain a nutrient reservoir from their dead and lysed neighboring cells. This might therefore be an early differentiation and apoptotic mechanism. PMID- 16213677 TI - Gas vesicles isolated from Halobacterium cells by lysis in hypotonic solution are structurally weakened. AB - Analysis of pressure-collapse curves of Halobacterium cells containing gas vesicles and of gas vesicles released from such cells by hypotonic lysis shows that the isolated gas vesicles are considerably weaker than those present within the cells: their mean critical collapse pressure was around 0.049-0.058 MPa, as compared to 0.082-0.095 MPa for intact cells. The hypotonic lysis procedure, which is widely used for the isolation of gas vesicles from members of the Halobacteriaceae, thus damages the mechanical properties of the vesicles. The phenomenon can possibly be attributed to the loss of one or more structural gas vesicle proteins such as GvpC, the protein that strengthens the vesicles built of GvpA subunits: Halobacterium GvpC is a highly acidic, typically "halophilic" protein, expected to denature in the absence of molar concentrations of salt. PMID- 16213678 TI - In situ analysis of native microbial communities in complex samples with high particulate loads. AB - In the present study a procedure combining a cell extraction method and Fluorescence In Situ Hybridization (FISH) for molecular monitoring and quantification of bacteria in soil and aquifer samples is presented. FISH was applied to bacterial cells extracted from the matrix by density gradient centrifugation. This separation method was applied to soil and aquifer samples and produced high cell recovery of 76.5%+/-4.4 and 78.0%+/-3.2, respectively. FISH, performed on the harvested cells, permitted a perfect visualization and quantification of bacteria. This approach is therefore promising for in situ detection of indigenous bacterial communities in complex samples. PMID- 16213679 TI - Libido and orgasm in middle-aged woman. AB - OBJECTIVES: Determine which social, demographic and sexual function variables that most influence libido or desire and orgasm domains in the premenopausal and postmenopausal women. METHODS: A cross-sectional analysis of 231 Colombian-born women, aged 40-62 years. The sexual function was measured by self-questionnaire. The analysis was performed by using the chi2-test and multivariate regression analysis. The sexual function was divided in five domains: desire, arousal, lubrication, orgasm, pain; additionally satisfaction was included. RESULTS: The women with a higher level of education and with a good perception of their satisfaction with their partners, reported better performance in the desire. Age and the non existence of sexual partner influenced in a negative way on the desire. In sexual active women the orgasm was negatively influenced by low satisfaction scores, lack of emotional closeness with their partners and low educational level. High scores in lubrication and desire were associated with a good performance in the orgasm. The hormone therapy (HT) was associated with better scores in orgasm. CONCLUSIONS: Age, level of education, the presence or lack of sexual partner, degree of satisfaction with emotional closeness with the partner and adequate lubrication, influence the desire and orgasm domains in a significant way. By identifying these associations we can then perform some inexpensive interventions. Improving lubrication for menopausal women. Including men in educational activities to sensitize them toward women's feelings. Organizing educational campaigns for middle-aged women to demystify that sexuality is only for young people. PMID- 16213680 TI - Predictive factors for age at menopause in Caucasian females. AB - OBJECTIVE: Early onset of menopause results in the premature exposure to low estrogen levels and is associated with a number of postmenopausal health problems and higher risk of mortality. The aim of this study was to determine genetic and environmental factors associated with age at natural and surgical menopause. METHODS: Multiple regression analysis using a sample of Caucasians composed of 154 females with surgical and 248 with natural menopause. RESULTS: Breastfeeding is a significant predictor of earlier natural menopause (P<0.05). Use of oral contraceptives and smoking were not significantly associated with age at menopause. Females who did not have history of pregnancies are at significantly higher risk (P<0.001) of getting early surgical menopause than those who did. We also tested the association of seven single nucleotide polymorphisms (SNPs) of the estrogen receptor alpha (ER-alpha) gene with age at menopause. No association was observed with age at menopause but the PvuII p allele was overrepresented in women with surgical menopause and associated with menopause per se (P=0.029; OR=1.8, 95% CI=1.1-3.0). CONCLUSIONS: Breastfeeding and alcohol consumption are significantly associated with earlier natural menopause. No significant effects of the ER-alpha genotypes were observed on the age of menopause. Given the important role of the ER-alpha in estrogen signaling, which directly influences the menopausal process, further studies are required to better define the relationship between this gene and age at menopause. PMID- 16213681 TI - Interfield differences in intensity and frequency representation of evoked potentials in rat auditory cortex. AB - Existing studies have demonstrated interfield differences in functional organizations and neuronal responsive properties at a single neuron level in the auditory cortex, suggesting complicated encoding of sound frequency and intensity. The objective of present work is, by characterizing cortical auditory evoked potentials (AEPs), to bridge neural characteristics between a single neuron and field levels and to identify the interfield differences in the auditory cortex specifically in terms of spatial representation, which will be useful in guiding future unit studies. The AEP mapping found that each of auditory fields, which could be identified by a different tonotopic representation, showed interfield differences in an intensity-dependent spatial change, amplitude, latency, and amplitude-SPL (sound pressure level) function. These results also showed that many aspects of cortical representation were based on the cochlear properties, yet some were inconsistent. The intensity-dependent shift of activation in AI paralleled the tonotopic axis, which was similar to the place code in cochlea, while the shift in AAF and VAF did not parallel. Nevertheless, the amplitude-SPL function suggested that an underlying mechanism of all these shifts can be a compressive nonlinearity to CF tone, which is possibly formed in the cochlea and still preserved in the cortex. These results suggest that each field modifies the representation to handle a different aspect of sound information, which can be better analyzed than the cochlear representation. PMID- 16213682 TI - Anti-diarrheal effect of Galla Chinensis on the Escherichia coli heat-labile enterotoxin and ganglioside interaction. AB - Enterotoxigenic Escherichia coli (ETEC) is the most frequently isolated enteropathogen, accounting for approximately 210 million diarrhea episodes annually. ETEC-induced diarrhea is initiated by the binding of B subunit of heat labile enterotoxin (LTB) to the ganglioside G(M1) on the surface of intestinal epithelial cell. Therefore, we evaluated the inhibitory effects of 297 Chinese medicinal herbs on the LTB and G(M1) interaction by G(M1)-enzyme-linked immunosorbent assay. Galla Chinensis extract (GCE) exhibited anti-LT-induced diarrheal effect in the patent mouse gut assay, with IC50 value of 4.7+/-1.3 mg/ml. GCE also inhibited the binding of LTB to G(M1), suggesting that GCE suppressed the LT-induced fluid accumulation by blocking the binding of LTB to G(M1). Furthermore, the ethyl acetate (EA) soluble fraction was the most active fraction of Galla Chinensis that inhibiting the binding of LTB to G(M1) with an IC50 value of 153.6+/-3.4 microg/ml. The major components of the EA fraction should be phenolic derivatives according to a thin-layer chromatography analysis. Gallic acid, the major component of EA fraction, blocked the binding of LTB to G(M1), resulting in the suppression of LT-induced diarrhea. In conclusion, these data suggested that Galla Chinensis and gallic acid might be potent drugs for the treatment of LT-induced diarrhea. PMID- 16213683 TI - Hypocholesterolemic and antioxidant effects of aqueous extracts from the dried calyx of Hibiscus sabdariffa L. in hypercholesterolemic rats. AB - The present study was designed to investigate the hypolipidemic effects and antioxidant effects of Hibiscus sabdariffa L. (roselle) with regard to protection of LDL oxidation in vivo and ex vivo in rats made hypercholesterolemic by continuous cholesterol feeding. Administering the dried calyx extracts of roselle at doses of 500 and 1,000 mg/kg together with continuous cholesterol feeding to hypercholesterolemic rats for 6 weeks significantly decreased serum cholesterol level by 22% and 26%, respectively (p<0.001); serum triglycerides level by 33% and 28%, respectively (p<0.05); serum LDL level by 22% and 32%, respectively (p<0.05). However, serum HDL level was not affected. LDL was extracted from plasma of the hypercholesterolemic rats and the effects of the dried calyx extracts of roselle on the oxidation of LDL in vivo and ex vivo were examined. Six-week treatment with 250, 500 and 1,000 mg/kg of the extracts significantly decreased thiobarbituric acid reactive substances (TBARs) formation (p<0.05) while the formation of conjugated dienes during the oxidation of LDL induced by CuSO(4) was reduced, but not significantly different. These lines of evidence suggest that the aqueous extracts from the dried calyx of roselle possess both antioxidant effects against LDL oxidation and hypolipidemic effects in vivo. However, its mechanism(s) of action remains to be elucidated. PMID- 16213684 TI - Antimicrobial activities of the leaf extracts of two Moroccan Cistus L. species. AB - We used the standard M27-T technique to study organic and aqueous leaf extracts of two Moroccan Cistus L. species: Cistus villosus L. and Cistus monspeliensis L. (Cistaceae L.) used in traditional medicine, for their antimicrobial properties against microorganisms, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Candida krusei, Candida glabrata and Aspergillus fumigatus. The broth dilution method M27-T, standardized by the National Committee for Clinical Laboratory Standards (NCCLS) allowed to determine the minimum inhibitory concentrations (MICs) of different extracts. Results showed that the different extracts differed clearly in their antimicrobial activities. Cistus villosus extracts exhibited more interesting activity than Cistus monspeliensis extracts when used on Staphylococcus aureus (MIC=0.78 mg/ml) and Candida glabrata (MIC=0.19 mg/ml), which are the most susceptible microorganisms. On the other hand, Candida krusei and Aspergillus fumigatus were the least susceptible microorganisms to all Cistus extracts. Comparison results were carried out using chloramphenicol, amoxicillin and amphotericin B as standard antibiotics. PMID- 16213685 TI - Mechanisms of suppression of nitric oxide production by 3-O-methylquercetin in RAW 264.7 cells. AB - Rhamnus nakaharai Hayata (Rhamnaceae) is used as a folk medicine in Taiwan for treating constipation, inflammation, tumors, and asthma. 3-O-Methylquercetin (3 MQ), a main constituent of the plant, has been reported to have potential for use in the treatment of asthma. The mechanisms of anti-inflammation of 3-MQ are still unclear. Nitric oxide (NO) production induced by lipopolysaccharide (LPS) through iNOS expression in RAW 264.7 cells, a mouse macrophage cell line, may reflect the degree of inflammation and may provide a measure for assessing the effect of drugs on the inflammatory process. Therefore, we were interested in investigating the mechanisms of suppression of NO production by 3-MQ in RAW 264.7 cells. 3-MQ (1-10 microM) concentration-dependently inhibited LPS (100 ng/mL)-induced NO production in RAW 264.7 cells. The IC(50) value was calculated to be 4.23 microM. 3-MQ (1-10 microM) significantly and concentration-dependently inhibited LPS (100 ng/mL)-induced iNOS protein and mRNA expressions in cells. The IC(50) values were calculated to be 4.36 and 6.53 microM, respectively. There was no significant difference among these three IC(50) values of 3-MQ. In conclusion, 3-MQ may exert its anti-inflammatory effect through the inhibition of iNOS DNA transcription. PMID- 16213686 TI - Antioxidant and 15-lipoxygenase inhibitory activities of the Malian medicinal plants Diospyros abyssinica (Hiern) F. White (Ebenaceae), Lannea velutina A. Rich (Anacardiaceae) and Crossopteryx febrifuga (Afzel) Benth. (Rubiaceae). AB - The African flora contains numerous medicinal plants whose biological and chemical properties are incompletely known. Antioxidant and radical scavenging properties of plants are subject to intensive research. In the work described here, we have investigated the antioxidant activity of the plants Diospyros abyssinica (root bark), Lannea velutina (root bark and stem bark) and Crossopteryx febrifuga (seeds). Extracts of different polarity were assayed for radical scavenging activity, using the stable free radical diphenylpicrylhydrazyl, and for inhibition of enzymatic lipid peroxidation mediated by soybean 15-lipoxygenase. All plants investigated showed activity, but there were large differences between plants and between extracts. In general, Diospyros abyssinica and Lannea velutina were richer in antioxidants than Crossopteryx febrifuga. Lipophilic extracts were not active as radical scavengers, but did inhibit 15-lipoxygenase. Semipolar extracts (80% aqueous ethanol and methanol) of Diospyros abyssinica and Lannea velutina showed the highest activity both as radical scavengers and lipoxygenase inhibitors, and also gave the highest extract yields. These plants therefore appear to be excellent sources of antioxidants. PMID- 16213687 TI - Hepatoprotective and antioxidant effects of Hygrophila auriculata (K. Schum) Heine Acanthaceae root extract. AB - Hygrophila auriculata (K. Schum) Heine (syn. Asteracantha longifolia Nees, Acanthaceae) was widely used in the Indian systems of medicine for the treatment of various liver ailments. The hepatoprotective activity of the aqueous extract of the roots was studied on CCl(4)-induced liver toxicity in rats. The activity was assessed by monitoring the various liver function tests, viz. alanine transaminase, aspartate transaminase (AST), alkaline phosphatase (ALP), total protein and total bilirubin. Furthermore, hepatic tissues were subjected to histopathological studies. The root extract was also studied for its in vitro antioxidant activity using ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods. The extract exhibited significant hepatoprotective and antioxidant activities. PMID- 16213689 TI - Serotonin and dopamine transporter imaging in patients with obsessive-compulsive disorder. AB - In obsessive-compulsive disorder (OCD), the success of pharmacological treatment with serotonin re-uptake inhibitors and atypical antipsychotic drugs suggests that both the central serotonergic and dopaminergic systems are involved in the pathophysiology of the disorder. We applied [123I]-2beta-carbomethoxy-3beta-(4 idiophenyl)tropane (beta-CIT) and a brain-dedicated high-resolution single photon emission computed tomography (SPECT) system to quantify dopamine transporter (DAT) and serotonin transporter (SERT) availability. By comparing 15 drug-naive patients with OCD and 10 controls, we found a significantly reduced availability (corrected for age) of striatal DAT and of thalamic/hypothalamic, midbrain and brainstem SERT in OCD patients. Severity of OCD symptoms showed a significant negative correlation with thalamic/hypothalamic SERT availability, corrected for age and duration of symptoms. Our data provide evidence for imbalanced monoaminergic neurotransmitter modulation in OCD. Further studies with more selective DAT and SERT radiotracers are needed. PMID- 16213688 TI - Human muscle aging: ROS-mediated alterations in rectus abdominis and vastus lateralis muscles. AB - Aging is related to the accumulation of reactive oxygen species (ROS)-mediated oxidative damage. Considering the heterogeneity of age-related changes and the involvement of muscles in different functions, we compared the aging process in different functional muscles. We studied age-related changes in rectus abdominis (RA) and vastus lateralis (VL) in subjects of different age (18-48- and 66-90 year-old). We analysed fiber distribution, antioxidant enzymatic systems: Mn and CuZn superoxide dismutase (MnSOD, CuZnSOD), glutathione peroxidase (GSHPx), catalase (CAT), as well as oxidative damage markers: lipoperoxide levels (LPO), carbonylated proteins (CP), reduced and oxidized glutathione (GSH, GSSG) content and the GSH/GSSG ratio. In the muscles analysed, type I fiber increases during aging with a consequent decrease in type II distribution. In the elderly group RA MnSOD showed higher activity than VL. Furthermore, in RA MnSOD was higher in the elder group than in the younger group. CuZnSOD, as well as GSHPx and CAT activities remained unchanged. LPO levels in VL increase with age; moreover, in the elderly group VL showed higher value than RA. CP, GSH and GSSG remained unchanged, while GSH/GSSG decreases in RA during aging. In conclusion, a relationship between aging and ROS seems to exist, but oxidative processes could evolve in different ways in muscles with different functions. PMID- 16213690 TI - Quantifiable change in functional brain response to empathic and forgivability judgments with resolution of posttraumatic stress disorder. AB - Previous functional neuroimaging studies of posttraumatic stress disorder (PTSD) have mainly focused on symptom-provocation paradigms in combat-related PTSD. We sought to elucidate the effect of non-combat-related PTSD on the physiology of social cognition. Thirteen patients with PTSD underwent functional magnetic resonance imaging (fMRI) while they engaged in tasks that (i) involve speculation on another's intention, (ii) invoke empathy and (iii) involve making judgments of the forgivability of others' actions; each versus 'baseline' social reasoning judgments. A post-therapy fMRI scan followed a course of modified cognitive behavioural therapy. Post-therapy, we found increased activation in brain regions predicted on the basis of foregoing work in healthy subjects. These included significant left middle temporal gyrus activation in post-therapy response to empathy judgments and posterior cingulate gyrus activation in post-therapy response to forgivability judgments. The specific regions of the human brain activated by empathy and forgivability judgments changed with symptom resolution in PTSD. Time and therapy are likely contributory factors that lead to a degree of 'normalisation' of the neural response to these social cognition tasks. PMID- 16213691 TI - NMR studies of chloroform@cryptophane-A and chloroform@bis-cryptophane inclusion complexes oriented in thermotropic liquid crystals. AB - Oriented inclusion complexes of chloroform@cryptophane-A and chloroform@bis cryptophane were prepared using a nematic thermotropic liquid crystal (ZLI 1132), and the alignment and magnetic resonance properties of these host-guest systems were studied via (13)C NMR of the labeled guests. Large (1)H-(13)C dipolar splittings for (13)CHCl(3) guests indicated significantly enhanced (approximately 2-fold) ordering for the trapped vs. free ligands under all conditions studied, with similar ordering observed for the two complexes-despite significant differences in size and motional freedom between the hosts. For each environment, variable-temperature studies permitted the sign and magnitude of the order parameter for chloroform's C-H bond to be independently determined from the (13)C chemical shift anisotropy (CSA) shifts (via the gradient method) and the restored (1)H-(13)C dipolar couplings. In both systems, the results are consistent with overall alignment of the complexes such that the cage principal axis lies roughly perpendicular to the LC director. PMID- 16213692 TI - Case-based retrieval to support the treatment of end stage renal failure patients. AB - OBJECTIVE: In the present paper, we describe an application of case-based retrieval to the domain of end stage renal failure patients, treated with hemodialysis. MATERIALS AND METHODS: Defining a dialysis session as a case, retrieval of past similar cases has to operate both on static and on dynamic features, since most of the monitoring variables of a dialysis session are time series. Retrieval is then articulated as a two-step procedure: (1) classification, based on static features and (2) intra-class retrieval, in which dynamic features are considered. As regards step (2), we concentrate on a classical dimensionality reduction technique for time series allowing for efficient indexing, namely discrete Fourier transform (DFT). Thanks to specific index structures (i.e. k -d trees), range queries (on local feature similarity) can be efficiently performed on our case base, allowing the physician to examine the most similar stored dialysis sessions with respect to the current one. RESULTS: The retrieval tool has been positively tested on real patients' data, coming from the nephrology and dialysis unit of the Vigevano hospital, in Italy. CONCLUSIONS: The overall system can be seen as a means for supporting quality assessment of the hemodialysis service, providing a useful input from the knowledge management perspective. PMID- 16213693 TI - Mortality assessment in intensive care units via adverse events using artificial neural networks. AB - OBJECTIVE: This work presents a novel approach for the prediction of mortality in intensive care units (ICUs) based on the use of adverse events, which are defined from four bedside alarms, and artificial neural networks (ANNs). This approach is compared with two logistic regression (LR) models: the prognostic model used in most of the European ICUs, based on the simplified acute physiology score (SAPS II), and a LR that uses the same input variables of the ANN model. MATERIALS AND METHODS: A large dataset was considered, encompassing forty two ICUs of nine European countries. The recorded features of each patient include the final outcome, the case mix (e.g. age) and the intermediate outcomes, defined as the daily averages of the out of range values of four biometrics (e.g. heart rate). The SAPS II score requires 17 static variables (e.g. serum sodium), which are collected within the first day of the patient's admission. A nonlinear least squares method was used to calibrate the LR models while the ANNs are made up of multilayer perceptrons trained by the RPROP algorithm. A total of 13,164 adult patients were randomly divided into training (66%) and test (33%) sets. The two methods were evaluated in terms of receiver operator characteristic (ROC) curves. RESULTS: The event based models predicted the outcome more accurately than the currently used SAPS II model (P<0.05), with ROC areas within the ranges 83.9 87.1% (ANN) and 82.6-85.2% (LR) versus 80% (LR SAPS II). When using the same inputs, the ANNs outperform the LR (improvement of 1.3-2%). CONCLUSION: Better prognostic models can be achieved by adopting low cost and real-time intermediate outcomes rather than static data. PMID- 16213694 TI - Integrating regression formulas and kernel functions into locally adaptive knowledge-based neural networks: a case study on renal function evaluation. AB - OBJECTIVE: In many medical areas, there exist different regression formulas to predict/evaluate a medical outcome on the same problem, each of them being efficient only in a particular sub-space of the problem space. The paper aims at the development of a generic, incremental learning model that includes all available regression formulas for a particular prediction problem to define local areas of the problem space with their best performing formula along with useful explanation rules. Another objective of the paper is to develop a specific model for renal function evaluation using nine existing formulas. METHODS AND MATERIALS: We have used a connectionist neuro-fuzzy approach and have developed a knowledge-based neural network model (KBNN) which incorporates and adapts incrementally several existing regression formulas and kernel functions. The model incorporates different non-linear regression functions as neurons in its hidden layer and adapts these functions through incremental learning from data in particular local areas of the space. More specifically, each hidden neural node has a pair of functions associated with it--one regression formula, that represents existing knowledge and one Gaussian kernel function, that defines the sub-space of the whole problem space, in which the formula is locally adapted to new data. All these functions are aggregated and changed through incremental learning. The proposed KBNN model is illustrated using a medical dataset of observed patient glomerular filtration rate (GFR) measurements for renal function evaluation. In this case study, the regression function for each cluster is selected by the model from nine formulas commonly used by medical practitioners to predict GFR. 441 GFR data vectors from 141 patients taken from 12 sites in Australia and New Zealand have been used as a case study experimental data set. RESULTS: The proposed GFR prediction model, based on the proposed generic KBNN model, outperforms at least by 10% accuracy any of the individual regression formulas or a standard neural network model. Furthermore, we have derived locally adapted regression formulas to perform best on local clusters of data along with useful explanatory rules. CONCLUSION: The proposed KBNN model manifests better accuracy then existing regression formulas or neural network models for renal function evaluation and extracts modified formulas that perform well in local areas of the problem space. PMID- 16213695 TI - An incremental EM-based learning approach for on-line prediction of hospital resource utilization. AB - OBJECTIVE: Inpatient length of stay (LOS) is an important measure of hospital activity, health care resource consumption, and patient acuity. This research work aims at developing an incremental expectation maximization (EM) based learning approach on mixture of experts (ME) system for on-line prediction of LOS. The use of a batch-mode learning process in most existing artificial neural networks to predict LOS is unrealistic, as the data become available over time and their pattern change dynamically. In contrast, an on-line process is capable of providing an output whenever a new datum becomes available. This on-the-spot information is therefore more useful and practical for making decisions, especially when one deals with a tremendous amount of data. METHODS AND MATERIAL: The proposed approach is illustrated using a real example of gastroenteritis LOS data. The data set was extracted from a retrospective cohort study on all infants born in 1995-1997 and their subsequent admissions for gastroenteritis. The total number of admissions in this data set was n = 692. Linked hospitalization records of the cohort were retrieved retrospectively to derive the outcome measure, patient demographics, and associated co-morbidities information. A comparative study of the incremental learning and the batch-mode learning algorithms is considered. The performances of the learning algorithms are compared based on the mean absolute difference (MAD) between the predictions and the actual LOS, and the proportion of predictions with MAD < or = 1 day (Prop(MAD < or = 1)). The significance of the comparison is assessed through a regression analysis. RESULTS: The incremental learning algorithm provides better on-line prediction of LOS when the system has gained sufficient training from more examples (MAD = 1.77 days and Prop(MAD < or = 1) = 54.3%), compared to that using the batch-mode learning. The regression analysis indicates a significant decrease of MAD (p value = 0.063) and a significant (p-value = 0.044) increase of Prop(MAD < or = 1) with the incremental learning algorithm. CONCLUSIONS: The incremental learning feature and the self-adaptive model-selection ability of the ME network enhance its effective adaptation to non-stationary LOS data. It is demonstrated that the incremental learning algorithm outperforms the batch-mode algorithm in the on line prediction of LOS. PMID- 16213696 TI - A family based study of the genetic association between the PLA2G4D gene and schizophrenia. AB - The present study detected two single nucleotide polymorphisms (SNPs) at the PLA2G4D locus, rs2459692 and rs4924618, to investigate a genetic association between the PLA2G4D gene and schizophrenia. A total of 236 Chinese parent offspring trios of Han descent were recruited for the genetic analysis. The transmission disequilibrium test (TDT) did not show allelic association either for rs2459692 (chi(2) = 0.217, P = 0.641) or for rs4924618 (chi(2) = 0.663, P = 0.416). To see the combined effect of the PLA2G4D locus with the other three PLA2G4 genes, we applied the above two SNPs as a conditional marker to test the pair-wise combination for a disease association. The conditioning on allele (COA) test revealed a weak association for the rs2459692-PLA2G4A combination (chi(2) = 6.03, df = 2, P = 0.049), the rs2459692-PLA2G4B combination (chi(2) = 7.16, df = 3, P = 0.028) and the rs4924618-PLA2G4C combination (chi(2) = 7.01, df = 2, P = 0.03), whereas the conditioning on genotype (COG) test showed a weak association only for the rs4924618-PLA2G4C combination (chi(2) = 8.52, df = 3, P = 0.036). Because we performed a multi-locus analysis in this study, the weak association shown by the conditional tests could make little biological sense. In conclusion, the PLA2G4D gene may not be involved in a susceptibility to schizophrenia. PMID- 16213697 TI - Modification of eicosanoid profile in human blood treated by dual COX/LOX inhibitors. AB - The arachidonic acid metabolizing enzymes, the cyclooxygenases (COXs) and lipoxygenases (LOXs), have been implicated in the development of a variety of cancers and numerous new therapeutic inhibitors are currently under investigation. However, given the interdependence of the two pathways, the effect of inhibiting one pathway with relatively selective agents can only be appreciated in the in vivo situation. Clearly then, because of their potential beneficial or deleterious effects, it is important to understand the nature and levels of the resulting arachidonic acid metabolites when treating patients with relatively selective inhibitor drugs. In this study, using reference COX-2, 5-LOX and dual COX-2/5-LOX inhibitors, we devised a protocol which permitted the simultaneous quantification of eicosanoid metabolites formed during stimulation of human peripheral venous blood samples with the calcium ionophore, A23187, in the absence and presence of lipopolysaccharide (LPS). Not surprisingly, the end products of both COX and LOX pathways were affected depending on the inhibitor, or combination of inhibitors, used and the concentrations of drug tested. In conclusion, the method described permits the rapid screening of novel compounds for potentially positive and/or negative effects upon the products of arachidonic acid metabolism. PMID- 16213700 TI - Does dipyrone have any effect on respiratory function in COPD patients? AB - OBJECTIVE: Dipyrone (Novalgin) is an effective analgesic, antipyretic agent also with spasmolytic effects on various types of smooth muscles. It has recently been reported that dipyrone relaxes tracheal smooth muscle of guinea pig. In this present study, we aimed to investigate whether this and previously reported in vitro results have any consequences on the respiratory function of normal healthy volunteers and chronic obstructive pulmonary disease (COPD) patients. METHODS: In this one-centered, non-randomized, non-comparative, open labelled study, 15 normal healthy volunteers and 15 stable COPD patients, with partially reversible bronchospasm, diagnosed according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) criteria were enrolled in the study at the time they had any indication of dipyrone use. The spirometric tests were performed by a portable notebook and Medikro Spiro2000 spirometry programme-software 1.6 version, before 30, 60, 90, and 120 min after 20 mg/kg of orally dipyrone intake. Groups were compared with the General Linear Model Repeated Measures analysis of variance. RESULTS: None of the spirometric parameters evaluated showed any significant differences when compared with the baseline values in both groups. CONCLUSION: While dipyrone had no bronchodilator effects on either COPD patients or normal volunteers, it also did not impair the spirometric parameters. Since COPD is a disease characterized by a progressive and largely irreversible airflow limitation, dipyrone has no observable bronchodilator effect. However, since dipyrone does not impair the pulmonary function, it can be used safely in COPD patients when there is an indication. PMID- 16213702 TI - Redesigning a collection system for "small" consumer electronics. AB - This paper establishes that the collection system within the recycling scheme for consumer electronics in the Netherlands has not been entirely successful in convincing consumers to hand in their used appliances by means of dedicated collection routes. Particularly regarding small appliances, consumers are persisting in their habit of discarding their appliances by means of the refuse bag/bin for regular household waste. Therefore, the current collection system has been unable to direct consumer behavior in the desired direction. Consequently, the layout of the current system is reviewed and redesigned in order to tackle this problem. This design effort applies the Triad model (behavioral model) in the analysis stage and the Morphological Chart method (design tool) in the synthesis stage, and results in a concrete proposal for a new system. Finally, the applicability of this design approach beyond the specific circumstances of the case study presented in this paper is discussed. This case represents only one example of the broader, worldwide challenge to design appropriate collection systems to direct consumer behavior in desired directions. It is argued that the approach presented in this paper could be a valuable contribution to research dealing with this challenge. PMID- 16213701 TI - Exhaled breath condensate levels of 8-isoprostane, growth related oncogene alpha and monocyte chemoattractant protein-1 in patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) patients have increased neutrophils and macrophages in their lungs, and inflammation of the airway is related to oxidative stress. This study assessed the levels of 8-isoprostane (an oxidative stress marker) and chemokines related to neutrophil and monocyte inflammation (growth-related oncogene alpha [GROalpha] and monocyte chemoattractant protein-1 [MCP-1]) in the airway of ex-smoking COPD patients by exhaled breath condensate (EBC) collection. Thirty-two (28 males) stable COPD patients (14 with FEV(1) 50% [Group 1], 18 with FEV(1) <50% predicted [Group 2]) and 18 non-smoking age and sex-matched controls were studied in this cross-sectional study. EBC was collected using the EcoScreen (Jaeger, Germany) during 10 min of tidal breathing with the nose clipped. Concentrations of 8-isoprostane, GROalpha and MCP-1 were measured by enzyme immunoassays. COPD patients had a higher concentration of 8 isoprostane than controls (COPD versus control, P<0.001; Group 1 versus Group 2, P=0.045). 8-isoprostane increased across the groups from normal, Group 1 to Group 2 (r=0.64, P<0.001). The median intraquartile range (IQR) levels in pg/ml for GROalpha were 45.3(44.5-46.5), 45.4(44.5-46.0), 46.0(45.6-47.3), whereas MCP-1 levels were 5.3(5.2-5.9), 6.2(5.4-6.9) and 5.7(5.5-6.4) in Group 1, Group 2 COPD and control subjects, respectively. GROalpha level was lower in COPD patients when compared to controls (P=0.01). MCP-1 level did not differ between COPD and the control group. 8-isoprostane level, but not GROalpha and MCP-1, in EBC was increased in COPD patients with poorer lung function. This suggests an increased oxidative stress in the airway in patients with more severe COPD. PMID- 16213703 TI - Chromosome translocations in sarcomas and the emergence of oncogenic transcription factors. AB - A subset of sarcomas is characterised by recurrent chromosome translocations that generate novel fusion oncoproteins. One or both of the genes involved in these translocations often encode transcription factors, and the resulting fusion proteins have aberrant transcriptional function compared to their wild-type counterparts. These fusion transcription factors disrupt multiple biological pathways by altering expression of target genes, and thereby result in a variety of altered cellular properties that contribute to the tumourigenic process. However, experimental data indicate that the fusion gene alone is not sufficient for transformation in primary cells (EWS-FLI1) or tumourigenesis in the mouse (PAX3-FKHR, FUS-CHOP), suggesting that additional collaborating genetic alterations are required. In addition to improving our understanding of the etiology of these tumours, this accumulating knowledge of the oncogenic properties of these fusion proteins, their downstream targets, and cooperating genetic alterations will permit the development of a variety of novel approaches to improve the therapy of these cancers. PMID- 16213704 TI - ETS transcription factors and their emerging roles in human cancer. AB - Cancer can be defined as a genetic disease, resulting as a consequence of multiple events associated with initiation, promotion and metastatic growth. Cancer results from the loss of control of cellular homeostasis. Cell homeostasis is the result of the balance between proliferation and cell death, while cellular transformation can be viewed as a loss of relationship between these events. Oncogenes and tumour suppressor genes act as modulators of cell proliferation, while the balance of apoptotic and anti-apoptotic genes controls cell death. All cancer cells acquire similar sets of functional capacities: (1) independence from mitogenic/growth signals; (2) loss of sensitivity to "anti-growth" signals; (3) evade apoptosis; (4) Neo-angiogenic conversion; (5) release from senescence; and (6) invasiveness and metastasis. One of the goals of molecular biology is to elucidate the mechanisms that contribute to the development and progression of cancer. Such understanding of the molecular basis of cancer will provide new possibilities for: (1) earlier detection as well as better diagnosis and staging of disease with detection of minimal residual disease recurrences and evaluation of response to therapy; (2) prevention; and (3) novel treatment strategies. We feel that increased understanding of ETS-regulated biological pathways will directly impact these areas. ETS proteins are transcription factors that activate or repress the expression of genes that are involved in various biological processes, including cellular proliferation, differentiation, development, transformation and apoptosis. Identification of target genes that are regulated by a specific transcription factor is one of the most critical areas in understanding the molecular mechanisms that control transcription. Furthermore, identification of target gene promoters for normal and oncogenic transcription factors provides insight into the regulation of genes that are involved in control of normal cell growth, and differentiation, as well as provide information critical to understanding cancer development. This review will highlight the current understanding of ETS genes and their role in cancer. PMID- 16213705 TI - A lava rock-based biofilter for the treatment of alpha-pinene. AB - Biofiltration is an emerging technology in the United States that utilizes microorganisms to biodegrade harmful contaminants in air to carbon dioxide and water. Biofiltration is not only more cost effective, but also more environmentally friendly than traditional technologies such as thermal oxidation and chemical scrubbing. The primary objectives of the study were to operate a lava rock-based laboratory biofiltration system for the removal of alpha-pinene. A consortium of microorganisms to be used as an inoculum was recovered that was able to use alpha-pinene as a sole source of carbon and energy. The removal of alpha-pinene from the laboratory system was monitored with a total hydrocarbon analyzer (THA). Based on THA analysis, elimination capacities as high as 100+g/m(3)/h were obtained in the laboratory biofilters. Removal efficiencies averaged 99% over a two year period. The solid support maintained a neutral pH with no buffer addition throughout the two year study and microbial levels were maintained between 10(6) and 10(7) colony forming units (CFU)/g of solid support. Bacillus and Rhodococcus species were found to be the majority of the microorganisms in the biofilters over a two year period. This is the first time an organism from either of these genera has been reported to utilize alpha-pinene as a sole source of carbon and energy. Overall, a preselected consortium of microorganisms coupled with lava rock as a biofilter solid support achieved extended alpha-pinene treatment levels that far exceed previously published values. PMID- 16213706 TI - Synthesis and structure-activity relationships of novel benzene sulfonamides with potent binding affinity for bovine carbonic anhydrase II. AB - This manuscript reports the identification of a novel series of mono- and bis- benzene sulfonamides with potent binding affinity for bovine carbonic anhydrase II (bCAII). These compounds exhibited nanomolar equilibrium dissociation constants with K(i)'s ranging from 4.7 to 9.3nM. All compounds were ester derivatives of the weak affinity bCAII inhibitor, 4-carboxybenzenesulfonamide. Structure-activity relationships for this novel series of compounds are discussed. PMID- 16213707 TI - Geometric diversity through permutation of backbone configuration in cyclic peptide libraries. AB - Cyclic peptides offer the possibility of varying both scaffold geometry and R group functionality. For example, parameters such as ring size and the placement of D-amino acid and proline residues can have a dramatic effect on the conformations of cyclic peptides, allowing access to structurally diverse species based on simple modifications in their linear sequences. We synthesized a cyclic peptide library in which ring size, alpha-carbon stereochemistry, and proline placement were varied. Analysis of the products showed that heptapeptides in general cyclized more readily than hexapeptides, and within these groups the scaffolds with a greater number of pralines cyclized with markedly lower yields than scaffolds with fewer pralines. Split-pool libraries based on a sample set of these scaffolds showed that, in general, scaffold geometry outweighed side chains variation in determining cyclization efficiency. These concepts were applied to the synthesis of cyclodimeric variants of an inhibitor of actin assembly in Xenopus egg extracts, yielding side chain variants with improved potency over the original scaffold. PMID- 16213708 TI - Anti-malarial activity of Baylis-Hillman adducts from substituted 2 chloronicotinaldehydes. AB - New Baylis-Hillman adducts are synthesized based on substituted 2 chloronicotinaldehydes and screened for their in vitro anti-malarial activity against chloroquine sensitive and chloroquine resistant Plasmodium falciparum. Out of the six new compounds synthesized and screened, 2b, 2c and 2d compounds showed substantial anti-malarial activity. PMID- 16213709 TI - Indole-2-carboxamidines as novel NR2B selective NMDA receptor antagonists. AB - A novel series of indole-2-carboxamidine derivatives was prepared and identified as NR2B selective NMDA receptor antagonists. The influence of the substituents on the indole skeleton as well as the substitution of the benzyl moiety on the biological activity of the compounds was studied. Compound 5a was po active in the formalin test in mouse. PMID- 16213710 TI - Synthesis and preliminary biological evaluation of (2S,1'R,2'S)- and (2S,1'S,2'R) 2-(2'-phosphonocyclopropyl)glycines, two novel conformationally constrained l-AP4 analogues. AB - Two novel constrained l-AP4 analogues, (2S,1'R,2'S)- and (2S,1'S,2'R)-2-(2' phosphonocyclopropyl)glycines (7) and (8), were synthesized and evaluated as mGluR ligands. Compound 7 showed to be a group III mGluRs agonist with micromolar activity. PMID- 16213711 TI - Ketene aminal-based lactam derivatives as a novel class of orally active FXa inhibitors. AB - N,N'-Disubstituted ketene aminals are good bioisosteres of thiourea functional groups. We report the design and synthesis of a novel class of ketene aminal based lactam derivatives as potent and orally active FXa inhibitors. PMID- 16213712 TI - Modular polyketide synthases: Investigating intermodular communication using 6 deoxyerythronolide B synthase module 2. AB - A novel variant of 6-deoxyerythronolide B synthase (DEBS) module 2 was constructed to explore the balance between protein-protein-mediated intermodular channeling and intrinsic substrate specificity within DEBS. This construct, termed (N3)Mod2+TE, was co-incubated with a complementary, donor form of the same module, (N5)Mod2(C2), as well as with a mutant of (N5)Mod2(C2) with an inactive ketosynthase domain, in order to determine the extent of intermediate channeling versus substrate diffusion into the downstream module. PMID- 16213713 TI - Synthesis and in vitro anti-hepatitis B and C virus activities of ring-expanded ('fat') nucleobase analogues containing the imidazo[4,5-e][1,3]diazepine-4,8 dione ring system. AB - As part of our structure-activity relationship studies, we report here the synthesis and in vitro anti-HBV and anti-HCV activities of a number of ring expanded ('fat') nucleobases containing the imidazo[4,5-e][1,3]diazepine-4,8 dione ring system. One of the compounds, ZP-88, exhibited a good activity/toxicity profile against HBV by inhibition of the synthesis of extracellular virion release (EC(50)=1.7microM, CC(50)=286microM, SI=168) and intracellular HBV replication intermediates (EC(50)=8.4microM, CC(50)=286microM, SI=34) in cultured human hepatoblastoma 2.2.15 cells. By contrast, most of the compounds tested against HCV had only marginal activity/toxicity profile, although that was still better than that of the reference compound ribavirin. PMID- 16213714 TI - Photoreactive threading agent that specifically binds to abasic sites in DNA. AB - We report the synthesis and study of a photoreactive nitrobenzamide containing acridine that specifically interacts at abasic site in DNA by threading intercalation and introduces under irradiation a lesion on the opposite strand at the unpaired pyrimidine. PMID- 16213715 TI - Structural basis for the GSK-3beta binding affinity and selectivity against CDK-2 of 1-(4-aminofurazan-3yl)-5-dialkylaminomethyl-1H-[1,2,3] triazole-4-carboxylic acid derivatives. AB - A novel structural class of glycogen synthase kinase-3beta inhibitors is modeled using quantum mechanics, automated docking, and molecular dynamics simulations. The proposed binding modes identify important hydrogen bonds and salt-bridges with the ATP-binding pocket of the kinase. The modeled complexes justify the observed structure-activity relationships and provide a structural basis for the high selectivity of these inhibitors against cyclin dependent kinase-2. PMID- 16213716 TI - 3-(5-Chloro-2,4-dihydroxyphenyl)-pyrazole-4-carboxamides as inhibitors of the Hsp90 molecular chaperone. AB - Information from X-ray crystal structures of Hsp90 inhibitors bound to the human Hsp90 molecular chaperone was used to assist in the design of 3-(5-chloro-2,4 dihydroxyphenyl)-pyrazole-4-carboxamides as novel inhibitors of Hsp90. Accessing an extra interaction with the protein via Phe138 gave a significant increase in binding potency compared to similar analogues that do not make this interaction. PMID- 16213717 TI - Factors affecting the protease activity of venom from jellyfish Rhopilema esculentum Kishinouye. AB - In this paper, the effects of some chemical and physical factors such as temperature, pH values, glycerol, and divalent metal cations on the protease activity of venom from jellyfish, Rhopilema esculentum Kishinouye, were assayed. Protease activity was dependent on temperature and pH values. Zn(2+), Mg(2+), and Mn(2+) in sodium phosphate buffer (0.02M, pH 8.0) could increase protease activity. Mn(2+) had the best effects among the three metal cations and the effect was about 20 times of that of Zn(2+) or Mg(2+) and its maximal protease activity was 2.3x10(5)U/mL. EDTA could increase protease activity. PMSF had hardly affected protease activity. O-Phenanthroline and glycerol played an important part in inhibiting protease activity and their maximal inhibiting rates were 87.5% and 82.1%, respectively. PMID- 16213718 TI - New metabolically stable fatty acid amide ligands of cannabinoid receptors: Synthesis and receptor affinity studies. AB - We investigated the structure-activity relationships for the interactions of fatty acid amide analogs of the endocannabinoid anandamide with human recombinant cannabinoid receptors. Thirty-five novel fatty acid amides were synthesized using five different types of acyl chains and 11 different aromatic amine 'heads.' Although none of the new compounds was a more potent ligand than anandamide, we identified three amine groups capable of improving the metabolic stability of arachidonoylamides and their CB(1)/CB(2) selectivity ratio to over 20-fold, and several aromatic amines capable of improving the affinity of short chain or monosaturated fatty acids for cannabinoid CB(1) receptors. For the first time a tertiary amide of arachidonic acid was found to possess moderate affinity (K(i)=300 nM) for cannabinoid CB(1), but not CB(2), receptors. PMID- 16213719 TI - Specific sensing between inositol epimers by a bis(boronate). AB - Bis(boronates) that utilize internal photoinduced electron transfer (PET) quenching mechanisms can specifically signal the binding of chiro-inositol without responding to its epimer, myo-inositol. PMID- 16213720 TI - Conformationally restricted analogs of Combretastatin A-4 derived from SU5416. AB - A series of compounds originally derived from the vascular endothelial growth factor receptor tyrosine kinase inhibitor, SU5416, was synthesized and evaluated. The most potent compound in this series, compound 7, structurally resembles the potent anti-microtubule agent Combretastatin A-4, inhibited tubulin polymerization, and showed potent growth inhibitory activities on both prostate and breast cancer lines with IC(50) values in low to subnanomolar range. PMID- 16213721 TI - N-8-Substituted benztropinamine analogs as selective dopamine transporter ligands. AB - A series of N-8-substituted benztropinamines was synthesized and evaluated for binding at the dopamine (DAT), serotonin (SERT), norepinephrine (NET) transporters, and muscarinic M1 receptors. In general, the isosteric replacement of the C-3 benzhydrol ether of benztropine by a benzhydryl amino group was well tolerated at the DAT. However, for certain N-8 substituted derivatives, selectivity over muscarinic M1 receptor affinity was reduced. PMID- 16213722 TI - Identification and initial evaluation of 4-N-aryl-[1,4]diazepane ureas as potent CXCR3 antagonists. AB - The identification and evaluation of aryl-[1,4]diazepane ureas as functional antagonists of the chemokine receptor CXCR3 are described. Specific examples exhibit IC(50) values of approximately 60 nM in a calcium mobilization functional assay, and dose-dependently inhibit CXCR3 functional response to CXCL11 (interferon-inducible T-cell alpha chemoattractant/I-TAC) as measured by T-cell chemotaxis, with a potency of approximately 100 nM. PMID- 16213723 TI - In vitro photo-release of a TRPV1 agonist. AB - Intracellular photolysis of a novel 'caged' capsaicin analogue results in in vitro activation of the capsaicin receptor TRPV1. PMID- 16213724 TI - Selection of 5'-untranslated sequences that enhance initiation of translation in a cell-free protein synthesis system from wheat embryos. AB - Random libraries of mRNA 5'-leader sequences were screened to obtain some sequences that can stimulate the translation initiation in a cell-free translation system from wheat embryos as efficiently as the Omega sequence from tobacco mosaic virus. Several sequences that are as useful as the Omega sequence and are homologous to no known sequences survived the screening. We expect that these sequences add useful options to the cell-free protein synthesis system that is becoming a powerful tool in the post-genomic researches. PMID- 16213725 TI - Synthesis and preliminary evaluation of [3H]PSB-0413, a selective antagonist radioligand for platelet P2Y12 receptors. AB - The selective antagonist radioligand [(3)H]2-propylthioadenosine-5'-adenylic acid (1,1-dichloro-1-phosphonomethyl-1-phosphonyl) anhydride ([(3)H]PSB-0413) was prepared by catalytic hydrogenation of its propargyl precursor with a high specific radioactivity of 74Ci/mmol. In preliminary saturation binding studies, [(3)H]PSB-0413 showed high affinity for platelet P2Y(12) receptors with a K(D) value of 4.57nM. Human platelets had a high density of P2Y(12) receptors exhibiting a B(max) value of 7.66pmol/mg of protein. PMID- 16213726 TI - Synthesis and biological evaluation of novel C (7) modified chrysin analogues as antibacterial agents. AB - The natural product, chrysin (5,7-dihydroxy flavone), obtained from Oroxylum indicum, exhibits numerous biological activities including anticancer, anti inflammatory, and antiallergic activities. Three series of chrysin analogues were prepared, in which chrysin and heterocyclic moieties are separated by 3-carbon, 4 carbon, and 6-carbon spacers. All the derivatives were screened for antibacterial activity against a panel of susceptible and resistant Gram-positive and Gram negative organisms. It was observed that most of the derivatives displayed significant activity as compared to their parent compound (chrysin). PMID- 16213727 TI - Pharmacophoric features and Ca2+ ion holding capacity of verapamil. AB - Ab initio Hartree-Fock calculations have been performed at the 6-31G level to study the pharmacophoric features of verapamil. Both the unprotonated and the protonated forms of verapamil have been studied. The study predicts that the drug enters the body in protonated form and is anchored to the receptor via H-bond formation involving protonated amine. Huge conformational change as well as deprotonation is required before the drug is capable of holding Ca(2+) ions. Folded form of drug is capable of holding Ca(2+) ion and the chiral center also seems to be involved to certain extent. PMID- 16213728 TI - Synthesis, spectral studies and in vitro assessment for antiamoebic activity of new cyclooctadiene ruthenium(II) complexes with 5-nitrothiophene-2-carboxaldehyde thiosemicarbazones. AB - We report here the synthesis, characterization and in vitro antiamoebic activity of 5-nitrothiophene-2-carboxaldehyde thiosemicarbazones (TSC), 1-5, and their bidentate complexes [Ru(eta(4)-C(8)H(12))(TSC)Cl(2)] 1a-5a. The biological studies of these compounds were investigated against HK-9 strain of Entamoeba histolytica and the concentration causing 50% cell growth inhibition (IC(50)) was calculated in the micromolar range. The ligands exhibited antiamoebic activity in the range (2.05-5.29microM). Screening results indicated that the potencies of the compounds increased by the incorporation of ruthenium(II) in the thiosemicarbazones. The complexes 1a-5a showed antiamoebic activity with an IC(50) of 0.61-1.43microM and were better inhibitors of growth of E. histolytica, based on IC(50) values. The most promising among them is Ru(II) complex 2a having 1,2,3,4-tetrahydroquinoline as N(4) substitution. PMID- 16213729 TI - Metabolic conversion of 24-methyl-Delta25-cholesterol to 24-methylcholesterol in higher plants. AB - Feeding of chemically synthesized [27-13C]codisterol ([27-13C]2), [27-13C]24 epicodisterol ([27-13C]3), [23,24-2H2]codisterol ([23,24-2H2]2), and [26,27 2H6]24-methyldesmosterol ([26,27-2H6]8) to Oryza sativa cell cultures, followed by MS and NMR analysis of the biosynthesized dihydrobrassicasterol (9)/campesterol (10), revealed that both (24R)- and (24S)-epimers of 24-methyl Delta25-cholesterol (2/3) were converted to 9 and 10 via the common intermediate 24-methyldesmosterol (8). PMID- 16213730 TI - QSAR by LFER model of HIV protease inhibitor mannitol derivatives using FA-MLR, PCRA, and PLS techniques. AB - The present quantitative structure-activity relationship (QSAR) study attempts to explore the structural and physicochemical requirements of mannitol derivatives for HIV protease inhibitory activity using linear free energy related model of Hansch. QSAR models have been developed using electronic (Hammett sigma), hydrophobicity (pi), and steric (molar refractivity and STERIMOL L, B1, and B5) parameters of phenyl ring substituents of the compounds along with appropriate dummy variables. Whole molecular descriptors like partition coefficient (logP(calcd)) and molar refractivity (MR) were also tried as additional descriptors. Statistical techniques like stepwise regression, multiple linear regression with factor analysis as the data preprocessing step (FA-MLR), principal component regression analysis (PCRA), and partial least squares (PLS) analysis were applied to identify the structural and physicochemical requirements for HIV protease inhibitory activity. The generated equations were statistically validated using leave-one-out technique. The quality of equations obtained from stepwise, FA-MLR, PCRA, and PLS are of acceptable statistical range (explained variance ranging from 74.0% to 80.5%, while predicted variance ranges from 70.3% to 77.1%). The coefficient of molar refractivity shows that the activity decreases with increase in volume. Lipophilicity of the para substituents at Y position is conducive to the activity while lipophilicity of the para substituents at X position is detrimental to the activity. The coefficients of molar refractivity (mr(Y_p)) and STERIMOL parameters for para substituents at X and Y positions (B1(X_p) and B5(Y_p)) of the phenyl rings indicate that the width of the substituents at X position and the overall size of para substituents at Y position are the detrimental factors for the activity. The fluoro substituent at ortho position (Y) decreases the activity when compared to the corresponding unsubstituted congener. Presence of hydrogen bond donor groups at para position (Y) also reduces the activity. Additionally, presence of substituent at ortho position (X) and the presence of substituent at para position (Y) are conducive for the activity. Presence of fluorine at X and Y positions also increases the activity. PMID- 16213731 TI - Structure-activity relationships and enzyme inhibition of pantothenamide-type pantothenate kinase inhibitors. AB - A set of novel pantothenamide-type analogues of the known Staphylococcus aureus pantothenate kinase (SaPanK) inhibitors, N-pentyl, and N-heptylpantothenamide, was synthesized in three series. The first series of analogues (1-3) were designed as molecular probes of the PanK binding site to elucidate important structure-activity relationships (SAR). The second series of analogues (4-16) were designed using structural information obtained from the Escherichia coli PanK (EcPanK) structure by targeting the pantothenate binding site and the adjacent phenylalanine-lined lipophilic pocket. Insight into the antimicrobial effect of N-pentylpantothenamide (N5-Pan) through its conversion to the antimetabolite ethyldethia-CoA and further incorporation into an inactive acyl carrier protein analogue drove the development of the third series of analogues (17-25) to enhance this effect using substrate-like substitutions. Each of the analogues was screened for enzyme inhibition activity against a panel of pantothenate kinases consisting of EcPanK, Aspergillus nidulans (AnPanK), SaPanK, and the murine isoform (MmPanK1alpha). Series 1 demonstrated only modest inhibitory activity, but did reveal some important SAR findings including stereospecific binding. Series 2 demonstrated a much higher inhibition rate for the entire series and significant inhibition was seen with analogues containing alkyl substituents. Series 3 demonstrated the most preferential inhibition profile, with the highest inhibitory activity against the SaPanK and MmPanK1alpha. The MmPanK1alpha protein was inhibited by a broad spectrum of the compounds, whereas the E. coli enzyme showed greater selectivity. The overall activity data from these analogues suggest a complex and non-enzyme specific SAR for pantothenamide substrate/inhibitors of the different PanK enzymes. PMID- 16213732 TI - Synthesis and properties of 2'-O,4'-C-methyleneoxymethylene bridged nucleic acid. AB - A novel bridged nucleic acid (BNA) analogue, 2'-O,4'-C-methyleneoxymethylene bridged nucleic acid (2',4'-BNA(COC)), was synthesized and incorporated into oligonucleotides. The 2',4'-BNA(COC) modified oligonucleotides showed high binding affinity with an RNA complement and significant enzymatic stability against snake venom phosphodiesterase. PMID- 16213734 TI - Synthesis, in vitro cellular uptake and photo-induced antiproliferative effects of lipophilic hypericin acid derivatives. AB - Hypericin, a naturally occurring hydroxylated phenanthroperylene dione, is used as a powerful photosensitizer for photodynamic therapy as well as a diagnostic tool for the fluorescence detection of flat neoplastic lesions in the bladder of patients. Both applications are based on the tumouritropic characteristics of the compound. To get more insight into some of the physicochemical properties of hypericin affecting its tumouritropic characteristics, we set out to synthesize a series of more lipophilic hypericins. For this purpose, a synthetic pathway to hypericin acid amides with hydrocarbon chains of different lengths stably attached by an amide bond at position C10 was explored. Hypericin acid proved inert in amide forming reactions, whereas the precursor protohypericin acid showed higher reactivity and resulted in the desired amide derivatives, which afterwards can be easily converted into their phenanthroperylene dione form. Hexyl-, octyl-, decyl- and dodecylamides of hypericin acid were successfully synthesized in this way. In vitro cellular uptake and photo-induced antiproliferative effects of the compounds were evaluated, using the human moderately differentiated non-invasive papillary transitional carcinoma RT-112 cell line. Whereas the more lipophilic amides were taken up limitedly, the hexylamide accumulated approx. as well as hypericin itself. From the antiproliferative data it can further be concluded that not only the cellular uptake, but also the light-induced activity, is affected by the introduced structural changes. PMID- 16213733 TI - A QSAR study on influenza neuraminidase inhibitors. AB - Influenza is a major respiratory infection associated with significant morbidity in the general population and mortality in elderly and high-risk patients. It is an RNA virus that contains two major surface glycoproteins, neuraminidase and hemagglutinin. These proteins are essential for infection. Neuraminidase has been found to be a potential target to control influenza virus. In this paper, we have developed 17 quantitative structure-activity relationships (QSAR) for different sets of compounds to understand chemical-biological interactions governing their activities toward influenza neuraminidase. PMID- 16213735 TI - Synthesis and biological evaluation of phosphonated carbocyclic 2'-oxa-3'-aza nucleosides. AB - The synthesis of carbocyclic 2'-oxa-3'-aza-nucleosides has been described, based on the 1,3-dipolar cycloaddition of a new phosphonated nitrone with vinyl acetate followed by coupling with silylated nucleobases. The obtained compounds have been evaluated for their ability to inhibit the reverse transcriptase of avian myeloblastosis retrovirus: no significant activity has been observed. PMID- 16213736 TI - New high affinity H3 receptor agonists without a basic side chain. AB - In this study, we replaced the basic amine function of the known histamine H(3) receptor agonists imbutamine or immepip with non-basic alcohol or hydrocarbon moieties. All compounds in this study show a moderate to high affinity for the cloned human H(3) receptor and, unexpectedly, almost all of them act as potent agonists. Moreover, in the alcohol series, we consistently observed an increased selectivity for the human H(3) receptor over the human H(4) receptor, but none of the compounds in this series possess increased affinity and functional activity compared to their alkylamine congeners. In this new series of compounds VUF5657, 5-(1H-imidazol-4-yl)-pentan-1-ol, is the most potent histamine H(3) receptor agonist (pK(i) = 8.0 and pEC(50) = 8.1) with a 320-fold selectivity at the human H(3) receptor over the human H(4) receptor. PMID- 16213737 TI - QSAR study on para-substituted aromatic sulfonamides as carbonic anhydrase II inhibitors using topological information indices. AB - A linear quantitative structure-activity relationship has been developed for a series of para-substituted aromatic sulfonamides by using topological index methodologies. The compounds were studied for their carbonic anhydrase II (CAII) inhibitory activity. A large series of topological indices were calculated and the stepwise regression method was used to derive the most significant model. Very good results were obtained using multi-parametric regressions and showed that the information approach used in the present work is quite useful for modeling carbonic anhydrase inhibition. PMID- 16213738 TI - The lethal effect of bis-type azridinylnaphthoquinone derivative on oral cancer cells (OEC-M1) associated with anti-apoptotic protein bcl-2. AB - Several drugs of aziridinylbenzoquinone analogs have undergone clinical trials as potential antitumor drugs. These bioreductive compounds are designed to kill tumor cells preferentially within the hypoxic microenvironment. From our previous reported data, it was found that the synthesized 2-aziridin-1-yl-3-[(2-[2-[(3 aziridin-1-yl-1,4-dioxo-1,4-dihydronaphthalen-2 yl)thio]ethoxy]ethyl)thio]naphthoquinone (AZ-1) is a bioreductive compound with potent lethal effect on oral cancer cell, OEC-M1. It was found in this study that the lethal effect of the oral cancer cell lines OEC-M1 induced by AZ-1 was mediated through the cell cycle arrest and apoptosis pathway. The LC50 values of OEC-M1 and KB cells induced by AZ-1 compound were 0.72 and 1.02 microM, respectively, which were much lower than that of normal fibroblast cells (SF with LC50 = 5.6 microM) with more than 90% of normal fibroblasts surviving as compared to control at a concentration of AZ-1 as high as 2 microM. It was interesting to note that the LC50 of monotype diaziridinylbenzoquinone compound, diaziquone (AZQ), was 50 microM on OEC-M1 cells. Comparing the cytotoxicity of AZ-1 and AZQ on OEC-M1 cells, AZ-1 is approximately 70 times more potent than AZQ. By using Western blot, both G2/M phase cell cycle arresting protein, cyclin B, and anti apoptotic protein, bcl-2, were expressed in OEC-M1 cell when the concentrations of AZ-1 were increased from 0.125 to 0.5 microM and then decreased from 1 to 2 microM of AZ-1 treatment as compared with control for 24 h. Both proteins were expressed most abundantly at 0.5 microM AZ-1. However, the expression of bcl-2 protein in OEC-M1 was significantly decreasing in a dose-dependent manner and was only about 50% protein level at 2 microM AZ-1 for 48h as compared with control. The cell survival check protein p53 increased from 1.72- to 2.8-fold and 1.36- to 2.16-fold at concentrations of AZ-1 from 0.125 to 2.0 microM in a dose dependently increasing manner on OEC-M1 as compared with control for 24 and48 h treatments, respectively. The apoptotic-related phenomena were observed, which included apoptotic body formation and the enzyme activity change of caspase-3. The apoptotic bodies and caspase-3 activity of OEC-M1 were induced only at 2 microM AZ-1 for a 24h treatment, yet apoptotic body formation was observed at as low as 0.5 microM AZ-1 and in a dose-dependently increasing manner for a 48 h treatment. The caspase-3 activity was increased 20.6%, 26.8%, and 84.2%, respectively, at 0.5, 1, and 2muM concentrations of AZ-1 for a 48 h treatment as compared with control. These results indicate that AZ-1 induced the cell death of OEC-M1 through the G2/M phase arrest of cell cycle and anti-apoptosis first and then apoptosis following a 48 h treatment. All of the pathway might be associated with bcl-2 and p53 protein expression. We propose that the AZ-1 could be used as anti-oral cancer drug for future studies with animal models. PMID- 16213739 TI - Bcl-2 phosphorylation in the BH4 domain precedes caspase-3 activation and cell death after neonatal cerebral hypoxic-ischemic injury. AB - To date, there are very few in vivo studies addressing the role of Bcl-2 phosphorylation. In a model of neonatal hypoxic-ischemic (HI) brain injury, we characterized the spatial and temporal phosphorylation of Bcl-2 at serine-24 (PS24-Bcl-2), using a site-specific antibody. Very few cells positive for PS24 Bcl-2 were found in control animals, but the number increased during reperfusion in all investigated brain areas in the ipsilateral hemisphere after HI, particularly in the border region between intact and damaged tissue. The highest numbers were encountered 24 h post-HI. Phosphorylation of Bcl-2 at serine-24 coincided with cytochrome c release after hypoxia-ischemia and preceded caspase-3 activation. Injured neurons displayed a predominantly nuclear, but also mitochondrial, localization of PS24-Bcl-2 immunoreactivity. In conclusion, phosphorylation of Bcl-2 at serine 24 was induced by hypoxia-ischemia, presumably resulting in loss of its anti-apoptotic function. PMID- 16213740 TI - Evaluation of the effectiveness of treatment with erythropoietin on anemia, cognitive functioning and functions studied by comprehensive geriatric assessment in elderly cancer patients with anemia related to cancer chemotherapy. AB - The primary aim of the present study was to examine the relationship of changes in hemoglobin levels following recombinant human erythropoietin (rHuEPO) treatment to changes in cognitive functioning studied by Mini Mental State Examination (MMSE) in elderly cancer patients undergoing chemotherapy treatment. The secondary aim was that to assess the relationship of changes in hemoglobin levels following rHuEPO treatment to changes in functions studied by Comprehensive Geriatic Assessment (CGA), such as Activity of Daily Living (ADL), Instrumental Activities of Daily Living (IADL), Geriatric Depression Scale (GDS) and the Mini Nutritional Assessment (MNA). To this end, hemoglobin levels and cognitive functioning were evaluated in a sample of cancer patients prior to the start of chemotherapy treatment and again after 4, 8 and 12 weeks of treatment with chemotherapy plus rHuEPO. Ten elderly patients (mean age 71.4 years) were enrolled. At baseline, enrolled patients had a mean Hb value of 10.3g/dl. After 4 weeks of rHuEPO treatment, Hb values increased significantly (p<0.0001), with a mean increase of 1.2g/dl (range: 0.2-2.1). Remarkably, 8 out of 10 (80%) showed an increase of Hb levels >or=1g/dl in comparison to baseline and therefore were considered responders. At baseline, four patients (40%) showed a moderate cognitive impairment, whilst six patients (60%) showed a normal cognitive function. After 4 weeks of rHuEPO treatment nine patients (90%) showed a significant improvement of cognitive functions in comparison to baseline (p<0.005): eight of them were responders also to rHuEPO in terms of correction of anemia. The Spearman's rank correlation test showed a statistical significant correlation between Hb increase and increase in cognitive functioning assessed by MMSE after 4 weeks (p=0.049), 8 weeks (p=0.044) and 12 weeks (p=0.031) of rHuEPO treatment. Therefore, the findings of this study provide support for the hypothesis that significant increases in hemoglobin over the course of chemotherapy supplemented with rHuEPO administration would be accompanied by significant improvement in cognitive performance over the same interval. PMID- 16213741 TI - Allogeneic hematopoietic cell transplantation (HCT) following reduced-intensity conditioning in patients with acute leukemias. AB - The majority of patients with acute leukemia enter complete remission following induction therapy, but relapse despite consolidation and maintenance chemotherapy. Allogeneic hematopoietic cell transplantation (HCT) is the most effective consolidation therapy but unfortunately associated with high transplant related mortality (TRM). In order to decrease TRM but still apply a graft-versus tumor effect, allogeneic HCT protocols with reduced-intensity conditioning were developed and more than 5000 HCT, of which 1500 for acute leukemia, performed. Detailed information is available on more than 400 patients with acute leukemia. The results, summarized in this article, confirm that reduced-intensity preparative regimens lead to full donor chimerism and to generation of graft versus-leukemia (GvL) effects with curative potential in older patients (>60 years). Prospective-controlled clinical trials are needed in younger patients to compare results of HCT after reduced-intensity conditioning to those of HCT with conventional conditioning. PMID- 16213742 TI - Developments in ghrelin biology and potential clinical relevance. AB - The spiropiperidine, MK0677, has been exploited to characterize and expression clone the growth hormone secretagogue receptor (GHS-R). Cloning of this receptor led to identification of its natural ligands, ghrelin and adenosine. Targeted disruption of the Ghsr gene demonstrated unambiguously that the GH-releasing and orexigenic properties of ghrelin are dependent on Ghsr expression and that the orexigenic signal is mediated through neuropeptide Y and agouti-related peptide neurons. This review summarizes new developments in our understanding of the physiological roles of ghrelin and its receptor (GHS-R). Recent discoveries of the effects of ghrelin on the thymus and proinflammatory and chemotactic cytokine pathways stimulate renewed interest in potential clinical applications, which include age-associated disorders, such as metabolic disease, sarcopenia, congestive heart failure, atherosclerosis and anorexia. PMID- 16213744 TI - Pituitary radiotherapy: current controversies. AB - External beam radiotherapy has been used extensively in the management of patients with pituitary disease. However, in view of advances in the techniques of radiotherapy planning and administration, neurosurgery and pharmacological manipulation of the pituitary, there are a growing number of questions and controversies surrounding the current and future use of pituitary radiotherapy in the management of pituitary disease. PMID- 16213743 TI - Aldosterone: good guy or bad guy in cerebrovascular disease? AB - Stroke is a leading cause of disability in the Western world, yet the choices for therapeutic intervention are few. The complex role played by aldosterone in the pathogenesis of stroke is beginning to emerge. Chronic mineralocorticoid receptor (MR) blockade reduces the incidence of hemorrhagic strokes and the severity of damage caused by ischemic strokes. This appears to be a vascular phenomenon because MR blockade increases vessel lumen diameter, which presumably increases blood flow and perfusion of the tissue to reduce ischemic damage. However, the vascular protection afforded by MR antagonism is at odds with the results seen within the brain, where MR activation is required for neuronal survival. Both of these divergent effects have possible therapeutic implications for stroke. PMID- 16213745 TI - New developments in the treatment of postmenopausal breast cancer. AB - In recent years, new agents have challenged tamoxifen as the standard endocrine therapy for postmenopausal breast cancer. This article reviews developments with regard to the third-generation aromatase inhibitors (AIs)--anastrozole, letrozole and exemestane--and fulvestrant, the first of a new type of estrogen receptor antagonist that, unlike tamoxifen, has no partial agonist activity. The final results of the "Arimidex", Tamoxifen, Alone or in Combination (ATAC) trial, at a median follow-up of more than five years, and recent results from switching studies with anastrozole and exemestane, strengthen the position of these AIs as adjuvant treatment for hormone receptor-positive early breast cancer. Sequencing options for the future are also discussed because non-steroidal AIs are increasingly used early in the treatment sequence. PMID- 16213746 TI - Development of a sandwich ELISA assay for measuring bovine soluble type II IL-1 receptor (IL1R2) concentration in serum and milk. AB - The IL1R is composed of two kinds of molecule, type I (IL1R I) and type II (IL1R2). IL1R1 contributes to IL-1 signaling, whereas the IL1R2 has no signaling property and acts as a decoy for IL-1. In this study, we developed a bovine IL1R2 specific sandwich ELISA to examine the sIL1R2 concentration in serum and milk from dairy cows. The concentration of colostral IL-1beta was examined to estimate the correlation to sIL1R2. The results showed that the sIL1R2 concentration in sera and milk changes with the stages of lactation. The serum sIL1R2 concentrations were 5.56+/-0.69 ng/ml (colostrum), 3.14+/-0.72 ng/ml (the early stage of lactation) and 5.76+/-1.25 ng/ml (the late stage of lactation). The milk sIL1R2 concentrations were 1.83+/-0.47 ng/ml (colostrum), 0.73+/-0.22 ng/ml (the early stage of lactation) and 2.92+/-0.56 ng/ml (the late stage of lactation). The concentrations of IL1R2 in sera and milk were significantly higher at the late stage of lactation and colostrum than that of the early stage of lactation. The reduction rates of sIL1R2 levels from the colostrum to the early stage of lactation were 43.6% (serum) and 61% (whey). IL-1beta was detected in all the colostrum (995.9+/-346.6 ng/ml). Significant correlation was observed between the levels of colostral IL-1beta and IL1R2 (r=0.75). PMID- 16213747 TI - Cytokine-mediated modulation of leptin and adiponectin secretion during in vitro adipogenesis: evidence that tumor necrosis factor-alpha- and interleukin-1beta treated human preadipocytes are potent leptin producers. AB - Over the last decade, compelling evidence has been presented that cytokines affect adipocyte tissue formation and function. In this study we explored the effect of pro-inflammatory (i.e. interleukin (IL)-1beta, IL-6, interferon (IFN) gamma, and tumor necrosis factor (TNF)-alpha) versus anti-inflammatory cytokines (i.e. IL-4, IL-10, and transforming growth factor (TGF)-beta1) on leptin and adiponectin secretion during in vitro human adipogenesis. Confirmative to previous reports, conversion of precursor preadipocytes into mature adipocytes was completely inhibited upon exposure to TNF-alpha, IL-1beta, IFN-gamma, or TGF beta1. Hence, all these anti-adipogenic cytokines prevented release of adipocyte specific adiponectin. IFN-gamma also strongly reduced leptin production (> or =85%). However, TNF-alpha, IL-1beta, and TGF-beta1 stimulated leptin production from preadipocytes in the absence of mature adipocytes (20.6+/-5.4 ng/ml, 100.8+/ 18.2 ng/ml, and 5.4+/-0.4 ng/ml, respectively, compared to 6.6+/-0.8 ng/ml in control adipocyte cultures on day 21; n=4). IL-4, IL-6 and IL-10 did not, or only slightly, affect adipocyte differentiation and their hormonal secretion. In conclusion, adiponectin and leptin are both synthesized by adipocytes, whereas leptin is also produced by preadipocytes upon TNF-alpha or IL-1beta stimulation. These data suggest that preadipocytes could contribute more to total circulating leptin levels than has been previously considered, especially in diseased conditions were these pro-inflammatory factors play a prominent role. PMID- 16213748 TI - In vivo drug-response in patients with leukemic non-Hodgkin's lymphomas is associated with in vitro chemosensitivity and gene expression profiling. AB - Only a few approaches are available to address the mechanisms of cell death in vivo which are induced by anticancer treatment in patients with malignancies. In this study in vitro chemosensitivity testing of primary peripheral blood leukemic cells of five patients suffering from different leukemic non-Hodgkin's lymphomas was combined with the analysis of the in vivo rate of apoptosis by flow-cytometry (Annexin V and depolarisation of mitochondrial membrane potential (MMP) by JC-1). Furthermore, changes in expression patterns of apoptosis related proteins during chemotherapeutic treatment were detected by Western Blot. Gene expression profiling (HG-U133A, Affymetrix, Santa Clara, CA) was employed to identify common marker genes of in vivo drug response. In vitro chemosensitivity was tested using the cytotoxic agents which the patients were scheduled to receive and was strongly correlated with effective reduction of leukemic lymphoma cells in patients resulting in complete remissions in all five cases. Due to the rapid clearance of apoptotic tumor cells in vivo neither the analysis of the in vivo rate of apoptosis and depolarisation of MMP nor the assessment of expression of regulators of apoptosis showed concordant results concerning the drug response. However, assessment of gene expression during therapy could identify a set of 30 genes to significantly discriminate between samples from patients before treatment compared to samples from the same patients after receiving cytotoxic therapy. Among these 30 genes we found a high proportion of genes associated with apoptotic cell death, cell proliferation and cell cycle signalling including complement lysis inhibitor (clusterin/CLU), beta-catenin interacting protein (ICAT), peroxisome proliferator activated receptor alpha (PPARalpha), TNF alpha converting enzyme (ADAM17/TACE), homeo box A3 (HOX1), inositol polyphosphatase 5 phosphatase type IV (PPI5PIV) and inhibitor of p53 induced apoptosis alpha (IPIA Alpha/NM23-H6). These results indicate that in vitro chemosensitivity testing and gene expression profiling can successfully be utilised to analyse in vivo drug response in patients with leukemic NHL's and can be used to explore new pathway models of drug-induced cell death in vivo which are independent of different lymphoma subtypes and different treatment regimens. PMID- 16213749 TI - Chicken embryo related (CER) cell line for quantification of rabies neutralizing antibody by fluorescent focus inhibition test. AB - Levels of rabies virus neutralization antibody in sera from vaccinated dogs and cattle were either measured by mouse neutralization test (MNT) or by rapid fluorescent focus inhibition test (RFFIT), performed on CER monolayers. The two tests were compared for their ability to detect the 0.5 International Units/ml (I.U.) recommended by the World Health Organization (WHO) as the minimum response for proof of rabies immunization. A significant correlation was found between the two tests (n=211; r=0.9949 in dogs and 0.9307 in cows, p<0.001), good sensitivity (87.5%), specificity (94.7%) and agreement (96.6%) as well. RFFIT method standardized on CER cell system for neutralizing antibodies detection turns the diagnosis easier and less expensive, specially when a great number of samples must be tested from endemic areas as commonly found in Brazil. PMID- 16213750 TI - PEGylated murine Granulocyte-macrophage colony-stimulating factor: production, purification, and characterization. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) regulates proliferation, differentiation, and function of hematopoietic progenitor cells. Aside from expansion of hematopoietic cells, GM-CSF has shown efficacy in other diseases, including Crohn's disease. While GM-CSF being clinically used in humans, the ability to perform mechanistic studies in murine models is difficult due to the limited availability and rapid clearance of murine GM-CSF in the peripheral blood. To address these issues, we efficiently expressed murine GM-CSF under the control of the AOX1 gene promoter in Pichia pastoris using the Mut(S) strain KM71H. We describe the unique conditions that are required for efficient production by high-density fermentation and purification of mGM-CSF protein. Recombinant mGM-CSF protein was purified by tangential flow ultrafiltration and preparative reverse phase chromatography. To address limited half life or rapid clearance in mice, recombinant murine GM-CSF was modified by lysine-directed polyethylene glycol conjugation (PEGylation). PEG-modified and unmodified proteins were characterized by amino terminus sequence analysis and matrix assisted laser desorption ionization time-of-flight mass spectrometry. Under the mild reaction conditions, the recombinant protein is efficiently modified by PEGylation on an average of 2-3 sites per molecule. In vivo treatment of mice with PEGylated mGM-CSF, but not the unmodified recombinant mGM-CSF, reproduces the potent colony stimulating effects of human GM-CSF in patients on myeloid progenitor populations, as assessed by FACs analysis. This simplified approach for the expression, purification, and modification of a biologically potent form of murine GM-CSF should facilitate the study of central mechanisms of action in murine disease models. PMID- 16213751 TI - Effect of salinity on the immune response of tiger shrimp Penaeus monodon and its susceptibility to Photobacterium damselae subsp. damselae. AB - Addition of NaCl at 2.5% to tryptic soy broth (TSB) significantly increased the growth of Photobacterium damselae subsp. damselae. Tiger shrimp Penaeus monodon held in 25 per thousand seawater were injected with P. damsela subsp. damselae grown in TSB containing NaCl at 0.5%, 1.5%, 2.5% and 3.5% at a dose of 8.48 x 10(4)colony-forming units (cfu)shrimp(-1). Over 24-96 h, the cumulative mortality was significantly higher for the shrimp challenged with P. damselae subsp. damselae grown in 2.5% NaCl than those grown in 0.5%, 1.5% and 3.5% NaCl. In another experiment, P. monodon held in 25 per thousand were injected with TSB grown P. damselae subsp. damselae (8.48 x 10(4)cfushrimp(-1)), and then transferred to 5 per thousand, 15 per thousand, 25 per thousand (control) and 35 per thousand. After 96 h, the mortality was highest for the P. damselae subsp. damselae-injected shrimp held in 5 per thousand, and the lowest for the P. damselae subsp. damselae-injected shrimp held in 25 per thousand. In a separate experiment, P. monodon held in 25 per thousand and then transferred to 5 per thousand, 15 per thousand, 25 per thousand (control) and 35 per thousand were examined for immune parameters, and phagocytic activity and clearance efficiency of P. damselae subsp. damselae after 12-96 h. The THC, hyaline cell, phenoloxidase activity, respiratory burst, superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency decreased significantly for the shrimp held in 5 per thousand, 15 per thousand and 35 per thousand after 12h. It is concluded that tiger shrimp P. monodon transferred from 25 per thousand to low salinity levels (5 per thousand and 15 per thousand) and high salinity (35 per thousand) had reduced immune ability and decreased resistance against P. damselae subsp. damselae infection. PMID- 16213752 TI - Immunogenicity, retention and protective effects of the protein derivatives of formalin-inactivated red seabream iridovirus (RSIV) vaccine in red seabream, Pagrus major. AB - A formalin-inactivated virus was previously found to be efficient in protecting fish against challenge with red seabream iridovirus (RSIV), a DNA virus belonging to the Iridoviridae family. In the present study, we determined the amount of the virus in the vaccine in terms of the number of copies of the gene for the major capsid protein (MCP) gene by quantitative real-time PCR and examined the longevity and types of immune response generated after intramuscular vaccination. We also tested whether the protein components of the vaccine are able to mount a protective immune response in fish. The vaccine contained 10(7) MCP copies per microliter of vaccine, and was detected in blood, kidney and spleen of vaccinated fish up to 15 days post-vaccination. Fish vaccinated with either the intact formalin-inactivated vaccine or its protein derivatives had increased serum neutralization antibodies and enhanced expression of MHC class I, although the kinetics of expression varied among groups. However, only those vaccinated with the intact vaccine survived the virus challenge, and this indicates that serum neutralization antibodies have scarce role in protecting the fish against RSIV. We hypothesize that the cell-mediated immunity, particularly the MHC class I pathway is responsible for such protection. PMID- 16213753 TI - Genotype (cystatin C) and EEG phenotype in Alzheimer disease and mild cognitive impairment: a multicentric study. AB - Previous findings demonstrated that haplotype B of CST3, the gene coding for cystatin C, is a recessive risk factor for late-onset Alzheimer's disease (AD; Finckh, U., von der Kammer, H., Velden, J., Michel, T., Andresen, B., Deng, A., Zhang, J., Muller-Thomsen, T., Zuchowski, K., Menzer, G., Mann, U., Papassotiropoulos, A., Heun, R., Zurdel, J., Holst, F., Benussi, L., Stoppe, G., Reiss, J., Miserez, A.R., Staehelin, H.B., Rebeck, G.W., Hyman, B.T., Binetti, G., Hock, C., Growdon, J.H., Nitsch, R.M., 2000. Genetic association of the cystatin C gene with late-onset Alzheimer disease. Arch. Neurol. 57, 1579-1583). In the present multicentric electroencephalographic (EEG) study, we analyzed the effects of CST3 haplotypes on resting cortical rhythmicity in subjects with AD and mild cognitive impairment (MCI) with the hypothesis that sources of resting EEG rhythms are more impaired in carriers of the CST3 B haplotype than non carriers. We enrolled a population of 84 MCI subjects (42% with the B haplotype) and 65 AD patients (40% with the B haplotype). Resting eyes-closed EEG data were recorded in all subjects. EEG rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5-13 Hz), beta 1 (13-20 Hz), and beta 2 (20 30 Hz). EEG cortical sources were estimated by low-resolution brain electromagnetic tomography (LORETA). Results showed that the amplitude of alpha 1 (parietal, occipital, temporal areas) and alpha 2 (occipital area) was statistically lower in CST3 B carriers than non-carriers (P < 0.01). Whereas there was a trend towards statistical significance that amplitude of occipital delta sources was stronger in CST3 B carriers than in non-carriers. This was true for both MCI and AD subjects. The present findings represent the first demonstration of relationships between the AD genetic risk factor CST3 B and global neurophysiological phenotype (i.e., cortical delta and alpha rhythmicity) in MCI and AD subjects, prompting future genotype-EEG phenotype studies for the early prediction of AD conversion in individual MCI subjects. PMID- 16213754 TI - A molecular phylogeny of the Canidae based on six nuclear loci. AB - We have reconstructed the phylogenetic relationships of 23 species in the dog family, Canidae, using DNA sequence data from six nuclear loci. Individual gene trees were generated with maximum parsimony (MP) and maximum likelihood (ML) analysis. In general, these individual gene trees were not well resolved, but several identical groupings were supported by more than one locus. Phylogenetic analysis with a data set combining the six nuclear loci using MP, ML, and Bayesian approaches produced a more resolved tree that agreed with previously published mitochondrial trees in finding three well-defined clades, including the red fox-like canids, the South American foxes, and the wolf-like canids. In addition, the nuclear data set provides novel indel support for several previously inferred clades. Differences between trees derived from the nuclear data and those from the mitochondrial data include the grouping of the bush dog and maned wolf into a clade with the South American foxes, the grouping of the side-striped jackal (Canis adustus) and black-backed jackal (Canis mesomelas) and the grouping of the bat-eared fox (Otocyon megalotis) with the raccoon dog (Nycteruetes procyonoides). We also analyzed the combined nuclear+mitochondrial tree. Many nodes that were strongly supported in the nuclear tree or the mitochondrial tree remained strongly supported in the nuclear+mitochondrial tree. Relationships within the clades containing the red fox-like canids and South American canids are well resolved, whereas the relationships among the wolf-like canids remain largely undetermined. The lack of resolution within the wolf-like canids may be due to their recent divergence and insufficient time for the accumulation of phylogenetically informative signal. PMID- 16213755 TI - Phylogeography of the asp viper (Vipera aspis) inferred from mitochondrial DNA sequence data: evidence for multiple Mediterranean refugial areas. PMID- 16213756 TI - Stable isotopes, mass spectrometry, and molecular fluxes: applications to toxicology. AB - In order to meet the increasing demands for safe and affordable drugs, improvements in the efficiency and accuracy of every step in drug development are required. Accordingly, new approaches for assessing drug toxicity that are faster and more precise are in demand. Numerous approaches using -omics and systems biology are being developed to meet this demand and, while promising, they have not yet provided the improvements in toxicology promised. Other innovative methodologies for predicting and assessing toxicities should therefore be explored. Here we present a novel approach for directly measuring the in vivo response of specific metabolic pathways to toxic agents. Using stable isotopes and ultra sensitive mass spectrometry, the effect of an agent on myelin synthesis, protein synthesis, or cell proliferation can be directly measured. Examples are presented where this approach is used to detect toxicity in the liver, brain, peripheral neurons, breast, and skin. Collagen synthesis, microglia proliferation, myelin synthesis, tubulin synthesis, hepatic cell proliferation, epidermal cell proliferation and mammary epithelial cell proliferation are quantitatively determined in vivo, in a high throughput manner. This approach avoids the computationally complex approach of systems biology and allows the user to observe the emergent properties of the system directly and quantitatively. PMID- 16213758 TI - Most patients with varicose veins have fears or concerns about the future, in addition to their presenting symptoms. AB - INTRODUCTION: This study aimed to document fears and concerns of patients about their varicose veins. REPORT: Completed questionnaires about symptoms and concerns were brought to clinic by 62% (203/329) patients referred with uncomplicated primary varicose veins. 'Concerns, worries or fears' about their veins were reported by 79%. These were seldom mentioned in referral letters, and included fears about thrombosis (31%), trauma (16%), ulcers (15%) and general concerns about the future (57%). DISCUSSION: Many patients have unrecognised fears and concerns about their varicose veins. Specific enquiry and reassurance about these is part of good management and may avoid unnecessary treatment. PMID- 16213757 TI - The effects of clozapine on delayed spatial alternation deficits in rats with hippocampal damage. AB - Clozapine is an atypical antipsychotic drug that has been shown to improve spatial memory in some animal models; however its efficacy in reversing spatial memory impairment in rats with hippocampal lesions is unknown. To address this issue, we tested the effects of clozapine on delayed spatial alternation deficits in rats with hippocampal damage in three separate experiments. In each experiment, adult male rats received sham surgery or direct stereotaxic infusions of the excitotoxin, NMDA, into the hippocampus. In the first study, seven days after surgery, the sham control animals received daily saline injections while the lesioned animals were split into two groups that received daily saline or clozapine (2.0 mg/kg, sc) injections. During the fifth week of injections, all animals were tested in a food-motivated delayed spatial alternation task. Saline treated rats with excitotoxic hippocampal damage displayed significant deficits in delayed spatial alternation. Daily clozapine injections completely reversed this deficit. In a second experiment, it was found that clozapine treatment limited to the testing days only did not improve alternation performance in lesioned rats. Finally, in a third experiment, chronic clozapine treatment did not improve alternation performance in lesioned rats that were pre-trained in the alternation task prior to surgery. These results suggest that chronic, but not acute, clozapine treatment enables rats with hippocampal damage to develop new spatial learning, but can not rescue old spatial learning established prior to damage. These results may have implications for the treatment of cognitive deficits caused by hippocampal dysfunction in disorders such as schizophrenia, Alzheimer's disease, and others. PMID- 16213759 TI - Rotational diffusion of membrane proteins in aligned phospholipid bilayers by solid-state NMR spectroscopy. AB - Solid-state NMR experiments on mechanically aligned bilayer and magnetically aligned bicelle samples demonstrate that membrane proteins undergo rapid rotational diffusion about the normal in phospholipid bilayers. Narrow single line resonances are observed from 15N labeled sites in the trans-membrane helix of the channel-forming domain of the protein Vpu from HIV-1 in phospholipid bilayers with their normals at angles of 0 degrees, 20 degrees, 40 degrees, and 90 degrees, and bicelles with their normals at angles of 0 degrees and 90 degrees with respect to the direction of the applied magnetic field. This could only occur if the entire polypeptide undergoes rotational diffusion about the bilayer normal. Comparisons between experimental and simulated spectra are consistent with a rotational diffusion coefficient (DR) of approximately 10(5)s-1. PMID- 16213760 TI - Barnase thermal titration via molecular dynamics simulations: detection of early denaturation sites. AB - The thermal stability of barnase has been studied using constant pressure and temperature (CPT) molecular dynamics at different temperatures. Barnase X-ray coordinates were obtained from the Research Collaboratory for Structural Bioinformatics (RCSB) Protein Data Bank (PDB code:1rnb). Simulations were performed at 285, 295, 300, 335, 345, and 395 K in explicit water under periodic boundary conditions for 280 ps. For each simulation, conformations were saved every 0.2 ps. Root mean square deviation (RMSD) values were calculated relative to the starting structure at 300 K and at time t = 0. Root mean square fluctuation (RMSF) values were calculated relative to the average structure obtained from the 300K simulation. Both root mean square deviation and fluctuation analysis indicated the presence of discrete regions of hyper sensitivity along the barnase polypeptide chain. These regions exhibited spikes in flexibility prior to any global structural changes. The specific changes in barnase backbone flexibility are accompanied by increased phi/psi angle fluctuations. These results suggest the presence of early denaturation sites or denaturation nuclei whose local structure is disrupted prior to global structure disruption. Identification of denaturation nuclei suggests that appropriate amino acid replacements at these sites may lead to the design and development of more stable barnase mutants. This strategy of identifying denaturation nuclei in protein structures may represent a first step in the design of more stable protein structures. PMID- 16213761 TI - Regulation of lipolysis in isolated adipocytes of rainbow trout (Oncorhynchus mykiss): the role of insulin and glucagon. AB - In the present study, we have examined the effects of insulin and glucagon on the lipolysis of rainbow trout (Oncorhynchus mykiss). To this end, adipocytes were isolated from mesenteric fat and incubated in the absence (basal lipolysis) or presence of different concentrations of insulin and glucagon. In addition, to further elucidate the effects of these hormones in vivo on adipocyte lipolysis, both fasting and intraperitoneal glucagon injection experiments were performed. Basal lipolysis, measured as the glycerol released in the adipocyte medium, increased proportionally with cell concentration and incubation time. Cell viability was verified by measuring the release of lactate dehydrogenase (LDH) activity in the medium. Insulin (at doses of 35 and 350 nM) decreased lipolysis in isolated adipocytes of rainbow trout in vitro, while glucagon was clearly lipolytic at concentrations of 10 and 100 nM. Furthermore, hypoinsulinemia induced by fasting, as well as glucagon injection, significantly increased lipolysis in isolated adipocytes approximately 1.5- and 1.4-fold, respectively, when compared with adipocytes from control fish. Our data demonstrate that lipolysis, as measured in isolated adipocytes of rainbow trout, can be regulated by both insulin and glucagon. These results not only indicate that insulin is an important hormone in lipid deposition via its anti-lipolytic effects on rainbow trout adipocytes, but also reveal glucagon as a lipolytic hormone, as shown by both in vitro and in vivo experiments. PMID- 16213762 TI - Assessment of the influence of surgical technique on postoperative pain and wound tenderness in cats following ovariohysterectomy. AB - Elective ovariohysterectomy was performed on 66 cats. Surgical approach was flank (group F) or midline (group M) allocated by block randomisation. Pre-anaesthetic medication was acepromazine (0.1 mg/kg) via intramuscular injection. Anaesthesia was induced with intravenous thiopentone, and maintained with halothane in 100% oxygen. Carprofen (4 mg/kg) was administered by the subcutaneous route immediately after induction of anaesthesia. Postoperative pain and wound tenderness were assessed at 1, 3, 6, 9, 11-12 and 20-24h after the end of surgery, and the assessment outcome marked on visual analogue scales (VAS). Intervention analgesia (if pain VAS was >40 mm) was pethidine 4 mg/kg via intramuscular injection. Area under the curve (AUC) for VAS for pain and VAS for wound tenderness for each cat were calculated. AUC for wound tenderness was significantly greater for group F (P = 0.007). There was no significant difference for AUC for pain between the groups. In conclusion, wounds after flank ovariohysterectomy are significantly more tender than after midline ovariohysterectomy in the cat. This indicates that interactive methods, including wound palpation, must be used to assess postoperative pain and the findings should be appropriately weighted in the overall assessment. PMID- 16213763 TI - Virulent systemic feline calicivirus infection: local cytokine modulation and contribution of viral mutants. AB - Virulent systemic feline calicivirus (VS-FCV) is a novel, emerging pathogen with mortality up to 67% even in previously healthy adult cats; VS-FCV has resulted in at least six epidemics since 1998. Affected cats have systemic vascular compromise and hemorrhagic-fever like signs in part due to viral invasion of epithelium and endothelium, coupled with host cytokine responses. Affected skin tissues had, on average, 3.8 elevated cytokines compared with control tissue, with prominent upregulation in IL-10, TNF-alpha, and MIP-1alpha. Sequencing of most of the genomes of two VS-FCV strains documented patterns of virus relatedness and implicated changes in the capsid gene in the emerging phenotype, possibly through initiation of immune mechanisms manifest in the cytokine changes. Understanding the features contributing to the emergence of this disease is critical for management and prevention of this and similar outbreaks attributable to RNA viruses in animals and humans. PMID- 16213764 TI - Bilateral, hypertrophic neuritis of the brachial plexus in a cat: magnetic resonance imaging and pathological findings. AB - A 9-year-old Burmese cat was presented for investigation of a subacute onset of bilateral forelimb paresis. Magnetic resonance imaging of the cervico-thoracic vertebral column and brachial plexus revealed a bilaterally symmetrical, severe and diffuse swelling of the spinal nerves forming the caudal part of the brachial plexus. Histopathology of the abnormal nerve roots, spinal nerves and brachial plexi showed inflammatory and marked proliferative changes with similar features to that of hypertrophic neuritis of man. Hypertrophic neuritis in man is a rare, tumor-like, chronic inflammatory peripheral nerve disorder of unknown origin most frequently involving the brachial plexus. PMID- 16213766 TI - Natural FCoV infection: cats with FIP exhibit significantly higher viral loads than healthy infected cats. AB - Natural feline coronavirus (FCoV) infection has been shown to not only induce intestinal infection with viral shedding, but also systemic infection which either remains without clinical signs or leads to feline infectious peritonitis (FIP). As systemic infection is not the key event in the development of FIP, the question arises as to whether a potential difference in viral load might be of importance. Therefore, the purpose of this study was to quantitatively assess feline coronavirus (FCoV) RNA loads in haemolymphatic tissues of healthy, long term FCoV-infected cats and cats with FIP. In cats that died from FIP, viral loads were significantly higher, indicating a higher rate of viral replication or a reduced capacity for viral clearance in cats developing and/or suffering from FIP. PMID- 16213765 TI - Inhaled fluticasone reduces bronchial responsiveness and airway inflammation in cats with mild chronic bronchitis. AB - This study investigated the effect of inhaled fluticasone on lower airway inflammation and bronchial responsiveness (BR) to inhaled carbachol in cats with very mild, chronic bronchitis (n = 5) that were compared with healthy cats serving as controls (n = 6). Chest radiographs, BR tests performed non-invasively by barometric whole body plethysmography (BWBP) and bronchoalveolar lavage (BAL) were performed before and after treatment. BR was quantified by calculating the concentration of carbachol inducing bronchoconstriction (C-Penh300%), defined as a 300% increase of baseline Penh, an index of bronchoconstriction obtained by BWBP. BAL fluid was analyzed cytologically and the oxidant marker 8-iso-PGF2alpha was determined. At test 1, healthy cats and cats with bronchitis were untreated, whereas for test 2 inhalant fluticasone (250 microg once daily) was administrated for 2 consecutive weeks to cats with bronchitis. Control cats remained untreated. Inhaled fluticasone induced a significant increase in C-Penh300% and a significant decrease of BAL fluid total cells, macrophages, neutrophils and 8-iso PGF2alpha in cats with bronchitis, whilst untreated control cats did not show significant changes over time. This study shows that a 2-week fluticasone treatment significantly reduced lower airway inflammation in very mild bronchitis. BR could be successfully monitored in cats using BWPB and decreased significantly in response to inhaled fluticasone. 8-Iso-PGF2alpha in BAL fluid was responsive to treatment and appeared as a sensitive biomarker of lower airway inflammation in cats. PMID- 16213767 TI - Bone involvement in myelofibrosis: effectiveness of bisphosphonates. PMID- 16213768 TI - Tuberculosis of the shoulder joint. AB - OBJECTIVES: Skeletal tuberculosis is less common than the pulmonary form. The involvement of the shoulder joint is infrequent. We report our experience treating tuberculosis of the shoulder in 11 patients. METHODS: There were seven men and four women, ranging in age from 19 to 55 years (average 28.09 years). The duration of their complaints at presentation ranged from 3 to 24 months. The most common presentation was pain, which was seen in 10 joints. All of the patients had mild to moderate restriction of motion of the shoulder. On laboratory examination, the erythrocyte sedimentation rate was increased mildly. No patient had an active tuberculosis lesion or history of pulmonary disease. The diagnosis was based on the clinical picture and radiographic features, and was confirmed by open biopsy. The diagnosis was not confirmed by biopsy in one patient, but the family history and clinical and radiological features were highly suggestive of tuberculosis. Surgical debridement was done in two patients and open biopsy in eight patients in order to obtain samples for pathology. Arthrodesis was done in only one patient. In all patients, treatment began with a four-drug regimen for 2 months, followed by a two-drug regimen for 10 months. RESULTS: The mean follow-up period after the end of treatment was 28.72 months (range, 22-52 months). At the time of the last visit, all the lesions had healed without recurrence. Five cases had a painless, mobile shoulder, while three had mildly restricted shoulder motion without pain, and three had residual limitation of motion of the affected shoulder. CONCLUSIONS: Tuberculosis of the shoulder can be difficult to diagnose in the early stages. If not diagnosed early, bony tuberculosis may reduce the quality of life. Therefore, tuberculosis should be suspected in cases of long standing pain in the shoulder. It is necessary to keep tuberculosis in the differential diagnosis of several osseous pathologies. Arthrodesis should be reserved only for lesions that fail to heal after adequate chemotherapy and rehabilitation. PMID- 16213769 TI - Aromatase and regulation of bone remodeling. AB - Estrogens play a key role in regulating bone mineralization. Bone tissue expresses the enzymes that metabolize estrogens, as well as the alpha and beta receptors that mediate responses to estrogens. After the menopause, estrogen secretion by the ovaries is promptly replaced by production within tissues, which occurs chiefly via aromatization of adrenal steroids. Therefore, aromatase activity is a major determinant of estrogen activity in postmenopausal women. Studies are beginning to shed light on the mechanisms by which aromatase activity influences bone remodeling. PMID- 16213770 TI - Chronic inflammatory joint disease revealing borderline leprosy. AB - Musculoskeletal symptoms are not infrequent in leprosy and, when inaugural, may be difficult to differentiate from other conditions, most notably rheumatoid arthritis. We report the case of a 24 year-old man with a 5 year history of intermittent inflammatory arthritis and fever. Physical findings and radiographs were normal initially. Several years later, he had severe wasting of the hand muscles, stocking-glove sensory loss, burn scars on the hands, and plantar ulcers. Electrophysiological test results indicated sensory-motor neuropathy with predominant demyelination. Laboratory tests showed inflammation without immunological abnormalities. A prominent endoneurial inflammatory infiltrate composed of mononuclear cells was seen on a nerve biopsy specimen, suggesting leprosy. A family study then revealed that the patient's aunt had been diagnosed with leprosy. Dapsone, clofazimine, and rifampin were given. The joint manifestations and laboratory tests for inflammation improved. However, no changes were noted in the neurological symptoms. PMID- 16213771 TI - Is restless legs syndrome underrecognized? Current management. AB - Restless legs syndrome (RLS) is a poorly understood sensory-motor neurological disorder whose prevalence in Caucasian populations ranges from 10% to 15%. The patient reports unpleasant sensations in the lower limbs with dysesthesia resulting in an urge to move the legs. The symptoms occur during periods of inactivity, increasing in the evening and at night. Moving the legs provides relief. In 80% of cases, polysomnography shows periodic leg movements during sleep. Patients with idiopathic RLS often report similar symptoms in family members. Secondary RLS may be due to medications, diabetes mellitus, renal failure, iron deficiency, neurological disorders, or rheumatoid arthritis. In secondary RLS, the management rests on treatment of the cause. Symptomatic treatment is warranted in patients with moderate-to-severe symptoms that adversely affect the quality of life. Dopaminergic agents are tried first. When they fail or induce adverse effects, weak opioids, benzodiazepines, anticonvulsants or, if needed, strong opioids, may be used. PMID- 16213772 TI - The rheumatoid shoulder: current consensus on diagnosis and treatment. AB - Shoulder involvement is usually inconspicuous in patients with rheumatoid arthritis, and the clinical manifestations are nonspecific. Nevertheless, shoulder involvement should be sought routinely and detected early. Range of motion at the shoulder should be evaluated. Although normal radiographic findings do not rule out shoulder involvement, radiographs are crucial for detecting micro and macro-geodes during follow-up. The development of glenohumeral joint space narrowing is a turning point that indicates a risk of rapid joint destruction. Magnetic resonance imaging is useful for assessing the lesions and guiding the treatment strategy. Stepwise use of local interventions as indicated by imaging findings is recommended. Joint replacement should not be left too late, and surgical procedures on the shoulder should be built into the overall treatment plan. PMID- 16213774 TI - Oncogenic osteomalacia in a patient with hemangioma: a clinical diagnosis. PMID- 16213773 TI - Cutaneous lymphocytic vasculitis during TNFalpha antagonist therapy for polyarthritis. PMID- 16213775 TI - Population database on nine STR loci of the AmpFlSTR Profiler kit in Koreans. AB - Allele frequencies for nine STR loci included in the AmpFlSTR Profiler Kit (D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820), were obtained from a sample of 1206 unrelated individuals living in the central region of Korea. PMID- 16213776 TI - Forensic potential of the STR DXYS156 in Mexican populations: inference of X linked allele null. AB - The pentanucleotide STR (TAAAA)n DXYS156 offers advantages for genetic identity testing. In addition to establish the gender, DXYS156 expands the DNA profile and is able to indicate the possible geographic origin of the individual. We analyzed DXYS156 in 757 individuals of both sexes from Mexican populations. We studied the cosmopolitan Mestizo population and six Mexican ethnic groups: Tarahumaras, Purepechas, Nahuas, Mayas, Huicholes and Mezcala Indians. The six shorter (4-10) and the three larger alleles (11-13) were specific for the X and Y-chromosomes, respectively. A random distribution of alleles into genotypes was observed in males and females from each population. We estimated the power of exclusion for paternity testing according to the son's gender, and the power of discrimination in forensic casework. In addition, we detected a relatively high frequency of an X-linked allele null, principally in Mexican-Mestizos (3.6%), which must be considered when DXYS156 be applied for identification purposes. PMID- 16213777 TI - Emerging role of protein kinase B/Akt signaling in pancreatic beta-cell mass and function. AB - The serine-threonine kinase Akt also known as protein kinase B is one of the most studied molecules. In addition to the important role in carcinogenesis, Akt is a major regulator of carbohydrate metabolism. Akt mediates insulin-dependent glucose uptake in insulin-sensitive tissues. Recent evidence underscores the importance of Akt for regulation of beta-cell mass and function. This review summarizes current findings concerning the molecular mechanisms, downstream signaling pathways, and critical components involved in regulation of beta-cell mass and function by Akt. The results of these observations suggest that elucidation of critical downstream effectors of this signaling pathway could generate promising molecules as a potential target to induce proliferation and survival of beta-cells. PMID- 16213778 TI - Stem cell factor/c-Kit interactions regulate human islet-epithelial cluster proliferation and differentiation. AB - Stem cell factor (SCF), a progenitor cell growth factor, binds to and activates the c-Kit receptor tyrosine kinase, which is critical for early stem cell differentiation in haematopoiesis and gametogenesis. Nothing is known regarding these interactions during islet development in the human fetal pancreas. The present study was to investigate whether an increase in c-Kit receptor activity in isolated human fetal islet-epithelial clusters, by giving exogenous SCF, would promote beta-cell development. In the intact fetal pancreas, SCF and c-Kit were observed co-localizing with cytokeratin 19 in both ductal and newly forming islet cells. Islet cells isolated from 14 to 16 weeks fetal pancreata were cultured with SCF (50 ng/ml) or vehicle for 48 h. We observed an increase in the number of c-Kit-, pancreatic and duodenal homeobox gene 1- (PDX-1-), insulin- and glucagon expressing cells in the SCF-treated group (PDX-1 and insulin, p < 0.05). PDX-1 and c-Kit mRNA levels were also up-regulated in the SCF group (PDX-1, p < 0.05), with no change in preproinsulin or proglucagon gene expression. Co-localization of insulin with PDX-1 or c-Kit was observed frequently in SCF-treated cultures. A significantly (p < 0.05) greater proliferative capacity of islet-epithelial clusters was found in the SCF group in parallel with increased (p < 0.02) phosphorylation of Akt in a phosphatidylinositol-3 kinase (PI3K)-dependent manner. Our results demonstrate that SCF/c-Kit interactions are likely to be involved in mediating islet cell differentiation and proliferation during human fetal pancreatic development, and that phosphorylated Akt may have a role downstream of SCF/c-Kit signaling. PMID- 16213779 TI - Polymers from plants to develop biodegradable plastics. AB - Katrin Neumann et al. have recently shown that transgenic tobacco and potato plants can accumulate high levels of cyanophycin, a possible source for poly aspartate. This work opens the way to the future production of biodegradable plastics using a plant-based production system. Several problems need to be overcome first, such as growth retardation as a result of cyanophycin accumulating in the cytosol, and a co-production system needs to be developed for economical reasons. PMID- 16213780 TI - Evolution of diversity: the Cape flora. AB - Although the environmental correlates of plant species richness have long received attention, research into the genesis of this diversity is in its infancy. The recent development of molecular dating methods and the inference of robust phylogenetic hypotheses have made it possible to investigate this problem. I use the Cape flora as a model to show that much modern diversity could be the result of recruiting diverse lineages over the entire Cenozoic into this flora, followed by in situ diversification (thus increasing species richness) of at least some of these lineages over a long period in an environmentally heterogeneous area. PMID- 16213781 TI - A diffusion approach to labeling rows and columns in an irregular array. AB - A robust algorithm is presented for labeling rows and columns in an irregular array. The algorithm is based on hierarchical pattern matching to a local lattice, which is used as a template. Starting from the best local match, the pattern is expanded hierarchically to encompass the entire array. An application to labeling digitized images of an array of tissue sections mounted on a microscope slide is discussed. PMID- 16213782 TI - Computational models of working memory: putting long-term memory into context. AB - Detailed computational modeling of human memory has typically been aimed at either short-term (working) memory or long-term memory in isolation. However, recent research highlights the importance of interactions between these systems for both item and order information. At the same time, computational models of both systems are beginning to converge onto a common framework in which items are associated with an evolving "context" signal and subsequently compete with one another at recall. We review some of these models, and discuss a common mechanism capable of modelling working memory and its interaction with long-term memory, focussing on memory for verbal sequences. PMID- 16213783 TI - Empowering patients with essential information and communication support in the context of diabetes. AB - OBJECTIVES: Patients with diabetes need to be aware of essential information to be involved in decision-making, manage diabetes properly, and communicate with doctors and other healthcare providers effectively. We have developed Violet Technology (VT) to provide features beyond previous health information tailoring systems by dynamically prioritizing diabetes learning topics and providing integrated direct support for patient-provider communication through formulation of individualized agendas to take to healthcare encounters. METHODS: A particular feature of the VT approach is a Diabetes Information Profile (DIP) that models psychosocial and educational exposure features, as well as clinical characteristics, and considers expressed patient information preferences and recent information browsing history. The agenda facility recommends questions that the patients may have based on their profile, as well as helping to initialize a patient empowerment protocol. The technology uses a modular and extensible approach for key components, including consumer health information, prioritization rules, and methods of instantiating the DIP. VT has been implemented into a web portal for patient use. Two phases of evaluation studies have been conducted to collect patient and healthcare provider feedback. RESULTS AND CONCLUSIONS: Results indicate that VT prioritizes relevant and important information for individual patients. Moreover, both patients and providers indicate that formulating an agenda of questions is important for patients. More extensive system use is needed to establish if the technology can deliver an improved patient-provider partnership and, ultimately, improved health outcomes. PMID- 16213784 TI - Avian influenza A (H5N1). AB - Since their reemergence in 2003, highly pathogenic avian influenza A (H5N1) viruses have reached endemic levels among poultry in several southeast Asian countries and have caused a still increasing number of more than 100 reported human infections with high mortality. These developments have ignited global fears of an imminent influenza pandemic. The current knowledge of the virology, clinical spectrum, diagnosis and treatment of human influenza H5N1 virus infections is reviewed herein. PMID- 16213785 TI - Hepatitis B genotype and YMDD profiles in an untreated Irish population. PMID- 16213786 TI - Sleep breathing and periodic leg movement pattern in Angelman Syndrome: a polysomnographic study. AB - OBJECTIVE: The aim of this study was to evaluate the sleep breathing patterns and to detect the eventual presence of periodic leg movements (PLMs) in patients affected by Angelman syndrome (AS). METHODS: Ten children with AS were recruited to participate in the study; the clinical diagnosis was confirmed by the genetic analysis (maternal 15q deletion, uniparental paternal disomy, or mutation of the UBE3A gene). All patients but two had presented epileptic seizures. Two age matched groups of patients with mental retardation (MR) associated (MRE+) or not (MRE-) to epilepsy were used as control groups. All subjects underwent one polysomnographic recording, after one adaptation night. Sleep stages were scored according to standard criteria slightly modified in order to take into account the specific EEG patterns of AS, also the apnea/hypopnea index (AHI) was quantified; PLMs were identified and the PLM index (PLMI) was computed. The statistical analysis was carried out by means of the one-way ANOVA, followed by the Fisher LSD post-hoc test, when appropriate, and by means of the linear correlation coefficient between AHI and PLMI. RESULTS: Sleep macrostructure showed only few significant differences between children with AS and the other two groups of subjects: AS patients showed higher percentage of wakefulness after sleep onset and sleep onset latency; moreover, the percentage of REM sleep was reduced in AS and in MRE+ subjects. A tendency for AS subjects to present a higher PLMI than the other two groups was also found. AHI >5 was found in 30% of AS subjects, in 30.8% of MRE+, and only in 20% of MRE- patients (chi(2) = 2.359, NS); 70% of AS patients, 38.5% of MRE+, and 46.7% of MRE- subjects had PLMI >5 (chi(2) = 3.088, NS). CONCLUSIONS: These results confirm our previous questionnaire-based findings of a high prevalence of sleep breathing disorder and important PLMs in AS and allow us to hypothesize that epilepsy, rather than mental retardation, might exacerbate these sleep disorders. SIGNIFICANCE: Sleep breathing disorder and PLMs might contribute to the cognitive impairment and to the worsening of life quality of subjects with AS and with MR (mostly those with epilepsy). Therefore, our findings suggest the need to explore these sleep disorders in children affected by MR and to set up a correct treatment. PMID- 16213788 TI - Nursing as the cornerstone of effective supportive care for patients with cancer. PMID- 16213787 TI - People with intellectual disabilities and their need for cancer information. AB - Many people with intellectual disabilities will be affected by cancer in their lifetime. There is a lack of available knowledge about people with intellectual disabilities' understanding and experience of cancer, and about their need for cancer information. The principal aim of the study was to explore the information needs of people with intellectual disabilities who are affected by cancer. This was a qualitative, hypothesis generating study. Nine people with intellectual disabilities who were affected by cancer were identified using a purposive sampling method; five of these participated in the study. Data collection included observation of the use of a pictorial cancer information book designed for people with intellectual disabilities, and semi-structured interviews. Data analysis revealed three themes: (1) "Nobody told me": the desperate need for cancer information; (2) "That reminds me": telling my own story; and (3) "I don't know much about cancer": the difficulties of the staff supporting people with intellectual disabilities to read the cancer information book. The people with intellectual disabilities in this study were not provided with information about cancer. They wanted this information, and appeared to be capable of comprehending it. Further development of accessible information materials around cancer and palliative care is needed. PMID- 16213789 TI - Advances in medical therapy of inflammatory bowel disease. AB - The past few years have seen increased knowledge and optimism on the use of biological therapy in Crohn's disease. Important lessons have been learned from the expanding clinical experience with infliximab, fuelling belief that current treatment will continue to evolve as therapy is aimed at specific targets within the immune cascade. Several other studies of agents that target tumour necrosis factor-alpha revealed mixed and sometimes disappointing results. However, this was balanced by encouraging results with agents that inhibit lymphocyte trafficking, as well as with other biological agents. Previous disappointing results of biological therapy in ulcerative colitis have been overcome with recent positive clinical trials with infliximab and the anti-CD3 antibody visilizumab. There now appears to be an expanding array of treatment options available to clinicians; however, as the number of potential molecular targets expands, unanswered questions remain regarding optimal treatment strategies, the long-term safety of biologicals and the ability of these novel and often expensive therapies to alter the natural history of the disease. PMID- 16213790 TI - The proteasome in Alzheimer's disease and Parkinson's disease: lessons from ubiquitin B+1. AB - Ubiquitin-containing cellular inclusions are characteristic of major neurodegenerative diseases and suggest an involvement of the ubiquitin-proteasome system. The frameshifted form of ubiquitin has proved to be a valuable tool for studying the role of the ubiquitin-proteasome system. It is an endogenous reporter for proteasome activity in human pathology but it is also capable of inhibiting proteasomal degradation. Current studies have revealed that the frameshifted form of ubiquitin accumulates in the brains of patients with Alzheimer's disease but not in those with Parkinson's disease. PMID- 16213791 TI - Potential drug targets in Mycobacterium tuberculosis through metabolic pathway analysis. AB - The emergence of multidrug resistant varieties of Mycobacterium tuberculosis has led to a search for novel drug targets. We have performed an insilico comparative analysis of metabolic pathways of the host Homo sapiens and the pathogen M. tuberculosis. Enzymes from the biochemical pathways of M. tuberculosis from the KEGG metabolic pathway database were compared with proteins from the host H. sapiens, by performing a BLASTp search against the non-redundant database restricted to the H. sapiens subset. The e-value threshold cutoff was set to 0.005. Enzymes, which do not show similarity to any of the host proteins, below this threshold, were filtered out as potential drug targets. We have identified six pathways unique to the pathogen M. tuberculosis when compared to the host H. sapiens. Potential drug targets from these pathways could be useful for the discovery of broad spectrum drugs. Potential drug targets were also identified from pathways related to lipid metabolism, carbohydrate metabolism, amino acid metabolism, energy metabolism, vitamin and cofactor biosynthetic pathways and nucleotide metabolism. Of the 185 distinct targets identified from these pathways, many are in various stages of progress at the TB Structural Genomics Consortium. However, 67 of our targets are new and can be considered for rational drug design. As a case study, we have built a homology model of one of the potential drug targets MurD ligase using WHAT IF software. The model could be further explored for insilico docking studies with suitable inhibitors. The study was successful in listing out potential drug targets from the M. tuberculosis proteome involved in vital aspects of the pathogen's metabolism, persistence, virulence and cell wall biosynthesis. This systematic evaluation of metabolic pathways of host and pathogen through reliable and conventional bioinformatic methods can be extended to other pathogens of clinical interest. PMID- 16213792 TI - Parameter estimation in modulated, unbranched reaction chains within biochemical systems. AB - Modern biology is increasingly developing techniques for measuring time series of global gene expression and of many simultaneous proteins or metabolites. These data contain valuable information on the dynamics of cells, which has to be extracted with computational means. Given a suitable mathematical model, this extraction is in principle a straightforward regression task, but the complexity and nonlinearity of the differential equations that describe biological systems cause severe difficulties when the systems are of realistic size. We propose a method of stepwise regression that can be applied effectively to linear portions of pathways. The method may be combined with other estimation methods and either directly yields reasonable parameter estimates or at least provides appropriate start values for subsequent nonlinear search algorithms. We illustrate the method with the analysis of in vivo NMR data describing the dynamics of glycolytic metabolites in Lactococcus lactis. PMID- 16213793 TI - Folding degrees of azurins and pseudoazurins. Implications for structure and function. AB - A quantitative measure of the degree of folding of azurins and pseudoazurins has been made. We have found that the reduction potential of azurins and pseudoazurins is a function of the contribution to the degree of folding of His117, a key amino acid in electron transfer which is directly bonded to copper in these proteins. The folding degree of His117 explains 95% of the variance in the experimental values of the reduction potential of azurins and pseudoazurins. The change in the folding degree of this amino acid influences several geometric parameters of the main backbones of these proteins. Among them, the angle formed between N(His117)...Cu...S(Cys112), which plays an important role in electron transport, but not the N(His117)...Cu distance, shows some non-linear correlation with the reduction potential of azurins and pseudoazurins. However, it is only able to explain less than 75% in the variance of the reduction potential of these proteins instead of the 95% explained by the folding degree of His117. PMID- 16213794 TI - Using Nearest Feature Line and Tunable Nearest Neighbor methods for prediction of protein subcellular locations. AB - The subcellular location of a protein is closely correlated with it biological function. In this paper, two new pattern classification methods termed as Nearest Feature Line (NFL) and Tunable Nearest Neighbor (TNN) have been introduced to predict the subcellular location of proteins based on their amino acid composition alone. The simulation experiments were performed with the jackknife test on a previously constructed data set, which consists of 2,427 eukaryotic and 997 prokaryotic proteins. All protein sequences in the data set fall into four eukaryotic subcellular locations and three prokaryotic subcellular locations. The NFL classifier reached the total prediction accuracies of 82.5% for the eukaryotic proteins and 91.0% for the prokaryotic proteins. The TNN classifier reached the total prediction accuracies of 83.6 and 92.2%, respectively. It is clear that high prediction accuracies have been achieved. Compared with Support Vector Machine (SVM) and Nearest Neighbor methods, these two methods display similar or even higher prediction accuracies. Hence, we conclude that NFL and TNN can be used as complementary methods for prediction of protein subcellular locations. PMID- 16213795 TI - A proteome-wide analysis of domain architectures of prokaryotic single-spanning transmembrane proteins. AB - We performed a proteome-wide survey of the domain architectures in single spanning transmembrane (TM) proteins (single-spannings) from 87 sequenced prokaryotic (Bacterial and Archaean) genomes by assigning Pfam domains to their N tail and C-tail loops. Out of 14,625 single-spannings, 3,516 sequences have at least one domain assigned, and no domains were assigned to 7,850, with the remaining 3,259 with less reliable assignment. In the domain-assigned sequences, 3116 sequences are with at most two domains, and the other 400 sequences with more than two. The assigned domains distribute over 651 Pfam families, which account for 11.4% of the total Pfam-A families. Among the 651 families are mostly soluble-protein-originated ones, but only 21 families are unique to TM proteins. The occurrence frequency of the individual domain families follows a power-law, that is, 264 families occur only once, 106 just twice, and the families appeared more than 30 times are counted by only 39. It is found that the great majority of the sequences having one or two domains are of the type II topology with the C tail loop containing domains on it. On the contrary, the N-tail loop of the same type topology seldom carries domains. Importantly, the assigned domains are always found on the tail loops longer than 60 residues, even for the small domains with less than 30 residues. There are still as many as 5,800 sequences without assigned domains in spite of having at least one long tail, on which no less than 1,000 novel domain families are expected most likely to lie concealed unknown yet. We also investigated the domain arrangement preference and the domain family combination patterns in 'singlets' (single-spannings with one assigned domain) and 'doublets' (with two domains). PMID- 16213796 TI - Fungal metabolite gliotoxin blocks mast cell activation by a calcium- and superoxide-dependent mechanism: implications for immunosuppressive activities. AB - Fungal secondary metabolites such as gliotoxin, an epipolythiodioxopiperazine toxin produced by pathogenic fungi like Candida and Aspergillus, possess immunosuppressive activities and have been thought to contribute to pathology of fungal infections in animals and humans. Since recent studies show that mast cell plays a crucial role in the front of host defense, we examined whether fungal secondary metabolites affected mast cell activation. We found that gliotoxin had suppressive effects on FcepsilonRI-dependent or -independent mast cell activation, including degranulation, leukotriene C4 secretion, and TNF-alpha and IL-13 production. Gliotoxin also suppressed intracellular Ca2+ rise through store operated Ca2+ channels with a minimal effect on depletion of internal Ca2+ stores. Finally, gliotoxin induced intracellular production of superoxide possibly through a thiol redox cycling, which appeared to mediate suppressive effects on mast cell activation. These findings suggest that suppression of mast cell activation might contribute to the establishment of infections with gliotoxin-producing fungi. PMID- 16213798 TI - Projections to the alimentary canal from the dopaminergic neurons in the dorsal motor nucleus of the vagus of the rat. AB - The motility of the alimentary canal is regulated not only by neurons that contain acetylcholine or adrenaline, but also by nonadrenergic noncholinergic neurons. There are many neurons containing dopamine in the dorsal motor nucleus of the vagus (DMV). We examined the projections of these dopaminergic neurons to the alimentary canal with double-labeling immunohistochemistry for tyrosine hydroxylase (TH) and the retrograde tracer cholera toxin subunit b following its injection into the subdiaphragmatic esophagus, the cardia, the pylorus, the duodenum, the jejunum, and the ascending colon. Almost all double-labeled neurons were found in the half of the DMV caudal to the area postrema. In the caudal half of the DMV, about 58% of the TH-immunoreactive neurons projected to the cardia, about 36% projected to the pylorus, and about 28% projected to the subdiaphragmatic esophagus. Only a few TH-immunoreactive neurons projected to the duodenum, the jejunum, or the ascending colon. As a whole, less than 10% of the neurons in the DMV that projected to the alimentary canal showed TH-like immunoreactivity. These results suggest that some of the dopaminergic neurons in the DMV might regulate the activities of the stomach and the subdiaphragmatic esophagus. PMID- 16213797 TI - Successful production of pseudotyped rAAV vectors using a modified baculovirus expression system. AB - Scalable production of rAAV vectors remains a major obstacle to the clinical application of this prototypical gene therapy vector. A recently developed baculovirus-based production protocol (M. Urabe et al., 2002, Hum. Gene Ther. 13, 1935-1943) found limited applications due to the system's design. Here we report a detailed analysis of the stability of the original baculovirus system components BacRep, BacVP, and transgene cassette-containing BacGFP. All of the baculovirus helpers analyzed were prone to passage-dependent loss-of-function deletions resulting in considerable decreases in rAAV titers. To alleviate the instability and to extend the baculovirus platform to other rAAV serotypes, we have modified both Rep- and Cap-encoding components of the original system. The modifications include a parvoviral phospholipase A2 domain swap allowing production of infectious rAAV8 vectors in vivo. Alternatively, an infectious rAAV8 (or rAAV5) vector incorporating the AAV2 VP1 capsid protein in a mosaic vector particle with AAV8 capsid proteins was produced using a novel baculovirus vector. In this vector, the level of AAV2 VP1 expression is controlled with a "riboswitch," a self-cleaving ribozyme controlled by toyocamycin in the "ON" mode. The redesigned baculovirus system improves our capacity for rAAV manufacturing by making this production platform more applicable to other existing serotypes. PMID- 16213799 TI - Discrimination and versatility in mismatch repair. AB - Evolutionarily-conserved mismatch-repair (MMR) systems correct all or almost all base-mismatch errors from DNA replication via excision-resynthesis pathways, and respond to many different DNA lesions. Consideration of DNA polymerase error rates and possible consequences of excess gratuitous excision of perfectly paired (homoduplex) DNA in vivo suggests that MMR needs to discriminate against homoduplex DNA by three to six orders of magnitude. However, numerous binding studies using MMR base-mispair-recognition proteins, bacterial MutS or eukaryotic MSH2.MSH6 (MutSalpha), have typically shown discrimination factors between mismatched and homoduplex DNA to be 5-30, depending on the binding conditions, the particular mismatches, and the DNA-sequence contexts. Thus, downstream post binding steps must increase MMR discrimination without interfering with the versatility needed to recognize a large variety of base-mismatches and lesions. We use a complex but highly MMR-active model system, human nuclear extracts mixed with plasmid substrates containing specific mismatches and defined nicks 0.15 kbp away, to measure the earliest quantifiable committed step in mismatch correction, initiation of mismatch-provoked 3'-5' excision at the nicks. We compared these results to binding of purified MutSalpha to synthetic oligoduplexes containing the same mismatches in the same sequence contexts, under conditions very similar to those prevailing in the nuclear extracts. Discrimination against homoduplex DNA, only two-to five-fold in the binding studies, increased to 60- to 230-fold or more for excision initiation, depending on the particular mismatches. Remarkably, the mismatch-preference order for excision initiation was substantially altered from the order for hMutSalpha binding. This suggests that post-binding steps not only strongly discriminate against homoduplex DNA, but do so by mechanisms not tightly constrained by initial binding preferences. Pairs of homoduplexes (40, 50, and 70 bp) prepared from synthetic oligomers or cut out of plasmids showed virtually identical hMutSalpha binding affinities, suggesting that high hMutSalpha binding to homoduplex DNA is not the result of misincorporations or lesions introduced during chemical synthesis. Intrinsic affinities of MutS homologs for perfectly paired DNA may help these proteins efficiently position themselves to carry out subsequent mismatch-specific steps in MMR pathways. PMID- 16213800 TI - Quantitative analysis of androst-4-ene-3,6,17-trione and metabolites in human urine after the administration of a food supplement by liquid chromatography/ion trap-mass spectrometry. AB - 6-OXO, a new nutritional supplement commercially available on the internet, is sold as an aromatase-inhibitor and contains androst-4-ene-3,6,17-trione as active ingredient. This anabolic steroid is a prohibited substance in sports. Androst-4 ene-3,6,17-trione is metabolised to androst-4-ene-6alpha-ol-3,17-dione and androst-4-ene-6alpha,17beta-diol-3-one. A fast, sensitive and accurate LC/MS method was developed and validated for the quantification of androst-4-ene-3,6,17 trione and its metabolites in urine. The method is capable of determining the stereochemical position of the hydroxy-group at C-6 of the metabolites and consists of a liquid-liquid extraction step with diethylether after enzymatic hydrolysis, followed by separation on a reversed phase column. Ionisation of the analytes is carried out using atmospheric pressure chemical ionisation. The limit of quantification of the method was 5 ng/mL for all compounds. The accuracy ranged from 14.8 to 1.3% for androst-4-ene-3,6,17-trione, 9.4 to 1.6% for androst 4-ene-6alpha-ol-3,17-dione and 4.1 to 3.2% for androst-4-ene-6alpha,17beta-diol-3 one in the range of 5-1000 ng/mL. Using this method androst-4-ene-6alpha-ol-3,17 dione was identified as a major urinary metabolite, whereas androst-4-ene 6alpha,17beta-diol-3-one as a minor metabolite. While the parent compound is predominantly excreted in conjugated form, both metabolites are solely excreted as conjugates. PMID- 16213801 TI - Structural stability of amyloid fibrils of beta(2)-microglobulin in comparison with its native fold. AB - Among various amyloidogenic proteins, beta(2)-microglobulin (beta2-m) responsible for dialysis-related amyloidosis is a target of extensive study because of its clinical importance and suitable size for examining the formation of amyloid fibrils in comparison with protein folding to the native state. The structure and stability of amyloid fibrils have been studied with various physicochemical methods, including H/D exchange of amyloid fibrils combined with dissolution of fibrils by dimethylsulfoxide and NMR analysis, thermodynamic analysis of amyloid fibril formation by isothermal calorimetry, and analysis of the effects of pressure on the structure of amyloid fibrils. The results are consistent with the view that amyloid fibrils are a main-chain-dominated structure with larger numbers of hydrogen bonds and pressure-accessible cavities in the interior, in contrast to the side-chain-dominated native structure with the optimal packing of amino acid residues. We consider that a main-chain dominated structure provides the structural basis for various conformational states even with one protein. When this feature is combined with another unique feature, template-dependent growth, propagation and maturation of the amyloid conformation, which cannot be predicted with Anfinsen's dogma, take place. PMID- 16213802 TI - Comments on cirrhosis reversal. PMID- 16213803 TI - Estrogen and androgen play distinct roles in bone turnover in male mice before and after reaching sexual maturity. AB - Aromatase is the sole enzyme which converts androgen into estrogen. We have reported that aromatase-knockout (ArKO) mice showed bone loss by increased bone resorption not only in female but also in male mice, suggesting essential roles of estrogen in bone metabolism in both sexes. However, loss of testicular androgen by orchidectomy (ORX) could induce bone loss in male mice. To clarify the relationship between estrogen and androgen in bone metabolism in male mice, 7 week-old ArKO mice were orchidectomized (ORX) to induce a double deficiency of estrogen and androgen. Bone loss in ORX/ArKO mice was more severe than that in ORX/wild-type and sham/ArKO mice because of advanced bone resorption, indicating that androgen and estrogen individually regulate bone mass by suppressing bone resorption in male mice after reaching sexual maturity. Cortical bone formation was elevated in sham/ArKO mice, but ORX did not influence cortical bone formation in the adult male mice. To examine the influence of androgen deficiency in weaning stage, 3-week-old wild-type mice were orchidectomized. Four weeks after operation, periosteal bone formation in the femur was markedly reduced in ORX mice. Since cortical bone in the same age of ArKO mice was normal, testicular androgen is indispensable for cortical bone formation especially at puberty in male mice. Therefore, estrogen and androgen may play distinct roles in bone turnover of male mice before and after reaching sexual maturity. PMID- 16213804 TI - Cryptochromes. PMID- 16213805 TI - Autism spectrum disorder. PMID- 16213806 TI - Chemical stimuli induce courtship dominance in Drosophila. PMID- 16213807 TI - Gender trading in a hermaphrodite. PMID- 16213808 TI - Opsins: evolution in waiting. AB - Complete vertebrate genome sequencing has revealed a remarkable stability and uniformity in the protein-coding gene set, which at first glance might suggest that gene duplication events are relatively rare. This may be a red herring, or at least a red cichlid, as the Lake Malawi cichlid fishes show rapid and extensive duplication and diversification of their retinal cone photoreceptor opsin genes. PMID- 16213809 TI - Axon pruning: C. elegans makes the cut. AB - Axon pruning has recently been described in the simple nervous system of the nematode Caenorhabditis elegans. Generating excess processes and pruning may be a phylogenetically conserved feature reflecting a flexibility to modify neural circuits. PMID- 16213810 TI - Intraflagellar transport: keeping the motors coordinated. AB - Intraflagellar transport is a conserved delivery system that services eukaryotic cilia and flagella. Recent work in the nematode Caenorhabditis elegans has identified proteins required for the functional coordination of intraflagellar transport motors and their cargoes. PMID- 16213811 TI - Evolutionary conflict: sperm wars, phantom inseminations. AB - A new experimental study has provided the first definitive evidence for conditional punishment of 'cheats' in a sperm-trading simultaneous hermaphrodite: the sea slug Chelidonura hirundinina. This also provides a rare unequivocal example of conditional reciprocity averting a 'tragedy of the commons' in biology. PMID- 16213812 TI - Plant meristems: mobile mediators of cell fate. AB - How do transcription factors control the fates of cells that express them? One class of plant transcription factors has recently been shown to function by regulating the synthesis of cytokinin and gibberellin hormones--mobile molecules more usually associated with long-distance signalling. PMID- 16213813 TI - Evolution: a study in bad taste? AB - Bitter tastes are among the most salient of life's experiences--who can forget one's first encounter with dandelion milk or a stout beer? Studies of the genes underlying these tastes are providing new perspectives on human origins and health. PMID- 16213814 TI - Sensory systems: fine-tuning the visual scene. AB - The visual system adjusts its properties for efficient representation of the objects present in the environment at the time. A new report suggests that complex processing of this sort can begin as early as the retina itself, but some important issues remain unresolved. PMID- 16213815 TI - Cell cycle: how cyclin E got its groove back. AB - CDK1 has long been known to orchestrate the passage of mammalian cells into and through mitosis. Recent work revisits the idea that CDK1, in conjunction with cyclin E, participates in S-phase entry as well. The new results shed light on a recent cell-cycle mystery, and provide another dramatic example of apparent functional redundancy among cyclins and cyclin-dependent kinases. PMID- 16213816 TI - The heparan sulfate proteoglycan syndecan is an in vivo ligand for the Drosophila LAR receptor tyrosine phosphatase. AB - BACKGROUND: Receptor tyrosine phosphatases (RPTPs) are essential for axon guidance and synaptogenesis in Drosophila. Each guidance decision made by embryonic motor axons during outgrowth to their muscle targets requires a specific subset of the five neural RPTPs. The logic underlying these requirements, however, is still unclear, partially because the ligands recognized by RPTPs at growth cone choice points have not been identified. RPTPs in general are still "orphan receptors" because, while they have been found to interact in vitro with many different proteins, their in vivo ligands are unknown. RESULTS: Here we use a new type of deficiency screen to identify the transmembrane heparan sulfate proteoglycan Syndecan (Sdc) as a ligand for the neuronal RPTP LAR. LAR interacts with the glycosaminoglycan chains of Syndecan in vitro with nanomolar affinity. Genetic interaction studies using Sdc and Lar LOF mutations demonstrate that Sdc contributes to LAR's function in motor axon guidance. We also show that overexpression of Sdc on muscles generates the same phenotype as overexpression of LAR in neurons and that genetic removal of LAR suppresses the phenotype produced by ectopic muscle Sdc. Finally, we show that there is at least one additional, nonproteoglycan, ligand for LAR encoded in the genome. CONCLUSIONS: Taken together, our results demonstrate that Sdc on muscles can interact with neuronal LAR in vivo and that binding to Sdc increases LAR's signaling efficacy. Thus, Sdc is a ligand that can act in trans to positively regulate signal transduction through LAR within neuronal growth cones. PMID- 16213817 TI - Generation of variants of a motor act in a modular and hierarchical motor network. AB - BACKGROUND: Most motor systems can generate a variety of behaviors, including categorically different behaviors and variants of a single motor act within the same behavioral category. Previous work indicated that many pattern-generating interneuronal networks may have a modular organization and that distinct categories of behaviors can be generated through flexible combinations of a small number of modules or building blocks. However, it is unclear whether and how a small number of modules could possibly generate a large number of variants of one behavior. RESULTS: We show that the modular feeding motor network of Aplysia mediates variations in protraction duration in biting-like programs. Two descending commands are active during biting behavior and trigger biting-like responses in a semiintact preparation. In the isolated CNS, when activated alone, the two commands produce biting-like programs of either long or short protraction duration by acting specifically on two modules that have opposite effects on protraction duration. More importantly, when coactivated at different frequencies, the two commands produce biting programs with an intermediate protraction duration. CONCLUSIONS: It was previously hypothesized that behavioral variants may be produced by combining different activity levels of multiple descending commands. Our data provide direct evidence for such a scheme and show how it is implemented in a modularly organized network. Thus, within a modular and hierarchical architecture, in addition to generating different categories of behavior, a small number of modules also efficiently implements variants of a single behavior. PMID- 16213818 TI - Arrestin1 mediates light-dependent rhodopsin endocytosis and cell survival. AB - BACKGROUND: Arrestins are pivotal, multifunctional organizers of cell responses to GPCR stimulation, including cell survival and cell death. In Drosophila norpA and rdgC mutants, endocytosis of abnormally stable complexes of rhodopsin (Rh1) and fly photoreceptor Arrestin2 (Arr2) triggers cell death, implicating Rh1/Arr2 bearing endosomes in pro-cell death signaling, potentially via arrestin-mediated GPCR activation of effector kinase pathways. In order to further investigate arrestin function in photoreceptor physiology and survival, we studied Arr2's partner photoreceptor arrestin, Arr1, in developing and adult Drosophila compound eyes. RESULTS: We report that Arr1, but not Arr2, is essential for normal, light induced rhodopsin endocytosis. Also distinct from Arr2, Arr1 is essential for light-independent photoreceptor survival. Photoreceptor cell death caused by loss of Arr1 is strongly suppressed by coordinate loss of Arr2. We further find that Rh1 C-terminal phosphorylation is essential for light-induced endocytosis and also for translocation of Arr1, but not Arr2, from dark-adapted photoreceptor cytoplasm to photosensory membrane rhabdomeres. In contrast to a previous report, we do not find a requirement for photoreceptor myosin kinase NINAC in Arr1 or Arr2 translocation. CONCLUSIONS: The two Drosophila photoreceptor arrestins mediate distinct and essential cell pathways downstream of rhodopsin activation. We propose that Arr1 mediates an endocytotic cell-survival activity, scavenging phosphorylated rhodopsin and thereby countering toxic Arr2/Rh1 accumulation; elimination of toxic Arr2/Rh1 in double mutants could thus rescue arr1 mutant photoreceptor degeneration. PMID- 16213819 TI - Mix and match color vision: tuning spectral sensitivity by differential opsin gene expression in Lake Malawi cichlids. AB - Cichlid fish of the East African Rift Lakes are renowned for their diversity and offer a unique opportunity to study adaptive changes in the visual system in rapidly evolving species flocks. Since color plays a significant role in mate choice, differences in visual sensitivities could greatly influence and even drive speciation of cichlids. Lake Malawi cichlids inhabiting rock and sand habitats have significantly different cone spectral sensitivities. By combining microspectrophotometry (MSP) of isolated cones, sequencing of opsin genes, and spectral analysis of recombinant pigments, we have established the cone complements of four species of Malawi cichlids. MSP demonstrated that each of these species predominately expresses three cone pigments, although these differ between species to give three spectrally different cone complements. In addition, rare populations of spectrally distinct cones were found. In total, seven spectral classes were identified. This was confirmed by opsin gene sequencing, expression, and in vitro reconstitution. The genes represent the four major classes of cone opsin genes that diverged early in vertebrate evolution. All four species possess a long-wave-sensitive (LWS), three spectrally distinct green sensitive (RH2), a blue-sensitive (SWS2A), a violet-sensitive (SWS2B), and an ultraviolet-sensitive (SWS1) opsin. However, African cichlids determine their spectral sensitivity by differential expression of primarily only three of the seven available cone opsin genes. Phylogenetic analysis suggests that all percomorph fish have similar potential. PMID- 16213820 TI - The interaction between binocular rivalry and negative afterimages. AB - Afterimage formation, historically attributed to retinal mechanisms, may also involve postretinal process. Consistent with this notion are results from experiments, reported here, investigating the interaction between binocular rivalry and negative afterimages (AIs). In Experiment 1, one eye was exposed to a grating never consciously experienced by the observer because this grating remained suppressed in rivalry throughout induction (the exclusively dominant stimulus was designed to preclude formation of an AI). As expected, the suppressed grating generated a vivid AI whose orientation could be accurately identified; not surprisingly, the strength of this AI varied with induction contrast. Experiment 2 revealed, however, that the strength of this AI produced during suppression was significantly weaker than the AI produced by that same stimulus when it was visible throughout the entire induction period, implying that some component of AI induction is susceptible to interocular suppression. In Experiment 3, AIs of dichoptic, orthogonally oriented gratings were induced in a way ensuring that one of the two gratings was exclusively dominant during the induction period. Dissimilar monocular AIs engaged in rivalry, as expected, but, surprisingly, the AI induced by the suppressed grating initially dominated. We offer two alternative accounts of this counterintuitive finding, both based on differential neural adaptation. PMID- 16213821 TI - Spatiotopic transfer of visual-form adaptation across saccadic eye movements. AB - Although conscious perception is smooth and continuous, the input to the visual system is a series of short, discrete fixations interleaved with rapid shifts of the eye. One possible explanation for visual stability is that internal maps of objects and their visual properties are remapped around the time of saccades, but numerous studies have demonstrated that visual patterns are not combined across saccades. Here, we report that visual-form aftereffects transfer across separate fixations when adaptor and test are presented in the same spatial position. The magnitude of the transsaccadic adaptation increased with stimulus complexity, suggesting a progressive construction of spatiotopic receptive fields along the visual-form pathway. These results demonstrate that basic shape information is combined across saccades, allowing for predictive and consistent information from the past to be incorporated into each new fixation. PMID- 16213822 TI - The DOCK180/Elmo complex couples ARNO-mediated Arf6 activation to the downstream activation of Rac1. AB - Cell motility requires extensions of the plasma membrane driven by reorganization of the actin cytoskeleton. Small GTPases, particularly the Rho family, are key regulators of this process. A second class of GTPases, the ADP-ribosylation factors (ARFs), have also been implicated in the regulation of the actin cytoskeleton and motility. ARF6 is intimately involved in the regulation of Rac activity; however, the mechanisms by which ARF activation leads to activation of Rac remain poorly understood. We have previously shown that expression of the ARF GEF ARNO in MDCK cells induces robust activation of Rac, the formation of large lamellipodia, and the onset of motility. We report here that ARNO-dependent activation of Rac is mediated by a bipartite Rac GEF, the Dock180/Elmo complex. Both DOCK180 and Elmo colocalize extensively with ARNO in migrating MDCK cells. Importantly, both a catalytically inactive Dock180 mutant and an Elmo mutant that fails to couple to Dock180 block ARNO-induced Rac activation and motility. In contrast, a similar mutant of the Rac GEF beta-PIX fails to inhibit ARNO-induced Rac activation or motility. Together, these data suggest that ARNO and ARF6 coordinate with the Dock180/Elmo complex to promote Rac activation at the leading edge of migrating cells. PMID- 16213824 TI - The tumor suppressor DAP kinase is a target of RSK-mediated survival signaling. AB - The viability of vertebrate cells depends on a complex signaling interplay between survival factors and cell-death effectors. Subtle changes in the equilibrium between these regulators can result in abnormal cell proliferation or cell death, leading to various pathological manifestations. Death-associated protein kinase (DAPK) is a multidomain calcium/calmodulin (CaM)-dependent Ser/Thr protein kinase with an important role in apoptosis regulation and tumor suppression. The molecular signaling mechanisms regulating this kinase, however, remain unclear. Here, we show that DAPK is phosphorylated upon activation of the Ras-extracellular signal-regulated kinase (ERK) pathway. This correlates with the suppression of the apoptotic activity of DAPK. We demonstrate that DAPK is a novel target of p90 ribosomal S6 kinases (RSK) 1 and 2, downstream effectors of ERK1/2. Using mass spectrometry, we identified Ser-289 as a novel phosphorylation site in DAPK, which is regulated by RSK. Mutation of Ser-289 to alanine results in a DAPK mutant with enhanced apoptotic activity, whereas the phosphomimetic mutation (Ser289Glu) attenuates its apoptotic activity. Our results suggest that RSK-mediated phosphorylation of DAPK is a unique mechanism for suppressing the proapoptotic function of this death kinase in healthy cells as well as Ras/Raf transformed cells. PMID- 16213823 TI - Xnf7 contributes to spindle integrity through its microtubule-bundling activity. AB - Regulation of microtubule dynamics and organization in mitosis by a number of microtubule-associated proteins (MAPs) is required for proper bipolar spindle assembly, yet the precise mechanisms by which many MAPs function are poorly understood. One interesting class of MAPs is known to localize to the nucleus during interphase yet fulfill important spindle functions during mitosis. We have identified Xenopus nuclear factor 7 (Xnf7), a developmental regulator of dorsal ventral patterning, as a microtubule-binding protein that also associates with the nuclear import receptor importin alpha/beta. Xnf7 localized to interphase nuclei and metaphase spindles both in Xenopus egg extracts and cultured cells. Xnf7-depleted spindles were hypersensitive to microtubule-depolymerizing agents. Functional characterization of Xnf7 revealed that it binds directly to microtubules, exhibits RING-finger-dependent E3-ubiquitin-ligase activity, and has C-terminal-dependent microtubule-bundling activity. The minimal microtubule bundling domain of Xnf7 was sufficient to rescue the spindle-hypersensitivity phenotype. Thus, we have identified Xnf7 as a nuclear MAP whose microtubule bundling activity, but not E3-ligase activity, contributes to microtubule organization and spindle integrity. Characterization of the multiple activities of Xnf7 may have implications for understanding human diseases caused by mutations in related proteins. PMID- 16213825 TI - Remapping auditory-motor representations in voice production. AB - Evidence regarding visually guided limb movements suggests that the motor system learns and maintains neural maps between motor commands and sensory feedback. Such systems are hypothesized to be used in a feed-forward control strategy that permits precision and stability without the delays of direct feedback control. Human vocalizations involve precise control over vocal and respiratory muscles. However, little is known about the sensorimotor representations underlying speech production. Here, we manipulated the heard fundamental frequency of the voice during speech to demonstrate learning of auditory-motor maps. Mandarin speakers repeatedly produced words with specific pitch patterns (tone categories). On each successive utterance, the frequency of their auditory feedback was increased by 1/100 of a semitone until they heard their feedback one full semitone above their true pitch. Subjects automatically compensated for these changes by lowering their vocal pitch. When feedback was unexpectedly returned to normal, speakers significantly increased the pitch of their productions beyond their initial baseline frequency. This adaptation was found to generalize to the production of another tone category. However, results indicate that a more robust adaptation was produced for the tone that was spoken during feedback alteration. The immediate aftereffects suggest a global remapping of the auditory-motor relationship after an extremely brief training period. However, this learning does not represent a complete transformation of the mapping; rather, it is in part target dependent. PMID- 16213827 TI - Functionally referential communication in a chimpanzee. AB - The evolutionary origins of the use of speech signals to refer to events or objects in the world have remained obscure. Although functionally referential calls have been described in some monkey species, studies with our closest living relatives, the great apes, have not generated comparable findings. These negative results have been taken to suggest that ape vocalizations are not the product of their otherwise sophisticated mentality and that ape gestural communication is more informative for theories of language evolution. We tested whether chimpanzee rough grunts, which are produced during feeding contexts, functioned as referential signals. Individuals produced acoustically distinct types of "rough grunts" when encountering different foods. In a naturalistic playback experiment, a focal subject was able to use the information conveyed by these calls produced by several group mates to guide his search for food, demonstrating that the different grunt types were meaningful to him. This study provides experimental evidence that our closest living relatives can produce and understand functionally referential calls as part of their natural communication. We suggest that these findings give support to the vocal rather than gestural theories of language evolution. PMID- 16213826 TI - Balancing selection and low recombination affect diversity near the self incompatibility loci of the plant Arabidopsis lyrata. AB - The self-incompatibility (S-) locus region of plants in the Brassica family is a small genome region. In Arabidopsis lyrata, the S-genes, SRK and SCR, encode the functional female and pollen recognition proteins, which must be coadapted to maintain correct associations between the two component genes, and thus self incompatibility (SI). Recombinants would be self-compatible and thus probably disadvantageous in self-incompatible species. Therefore, tight linkage between the two genes in incompatibility systems is predicted to evolve to avoid producing such recombinant haplotypes. The evolution of low recombination in S locus regions has not been rigorously tested. To test whether these regions' per nucleotide recombination rates differ from those elsewhere in the genome, and to investigate whether the A. lyrata S-loci have the predicted effect on diversity in their immediate genome region, we studied diversity in genes that are linked to the S-loci but are not involved in incompatibility and are not under balancing selection. Compared with other A. lyrata loci, genes linked to the S-loci have extraordinarily high polymorphism. Our estimated recombination in this region, from fitting a model of the effects of S-allele polymorphism on linked neutral sites, supports the hypothesis of locally suppressed recombination around the S locus. PMID- 16213829 TI - The social implications of embryo cryopreservation. AB - OBJECTIVE: To summarize the existing literature regarding the social implications of embryo cryopreservation and outline areas in need of further study. RESULT(S): The potential social impact of oocyte cryopreservation has not been investigated. Embryo cryopreservation has been increasingly used to improve the cost effectiveness of in vitro fertilization (IVF) and expand the options available to infertile couples, yet its widespread adoption has occurred more rapidly than our ability to study the social consequences for the couples and health professionals involved. For maintaining cryopreserved embryos, the existing literature is fragmented and incompletely explores the effects on an infertile couple's psychosocial health and personal relationships, their family planning strategies, or their preferences for the disposition of the embryos. Managing unclaimed embryos continues to create challenges for assisted reproduction professionals. CONCLUSION(S): We currently lack a thorough understanding of the numerous social implications of cryopreservation. Major areas for future research include the impact of stored embryos on couples' fertility intentions and psychosocial health, factors that affect couples' decisions about embryo disposition, strategies to minimize unclaimed embryos, and the consequences of oocyte/ovarian cryopreservation. PMID- 16213830 TI - Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test. AB - OBJECTIVE: To improve the sperm chromatin dispersion (SCD) test and develop it as a simple kit (Halosperm kit) for the accurate determination of sperm DNA fragmentation using conventional bright-field microscopy. DESIGN: Method development, comparison, and validation. SETTING: Medical genetics laboratory, academic biology center, and reproductive medicine centers. PATIENT(S): Male infertility patients attending the Reproductive Medicine Center. A varicocele patient and a group of nine fertile subjects. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): [1] The quality of chromatin staining in relaxed sperm nuclear halos and tail preservation; [2] SCD scoring reproducibility; [3] comparison with the sperm chromatin structure assay in 45 samples; [4] frequency of sperm with DNA fragmentation after incubation with increasing doses of the nitric oxide donor sodium nitroprusside and in sperm samples for 9 fertile men, 46 normozoospermic patients, 23 oligoasthenoteratozoospermic patients, and a subject with varicocele. RESULT(S): The sperm nuclei with DNA fragmentation, either spontaneous or induced, do not produce or show very small halos of DNA loop dispersion after sequential incubation in acid and lysis solution. The improved SCD protocol (Halosperm kit) results in better chromatin preservation, therefore highly contrasted halo images can be accurately assessed using conventional bright-field microscopy after Wright staining. Moreover, unlike in the original SCD procedure, the sperm tails are now preserved, making it possible to unequivocally discriminate sperm from other cell types. The chi2 test did not detect significant differences in the mean number of sperm cells with fragmented DNA as scored by four different observers. The intraobserver coefficient of variation for the estimated percentage of spermatozoa with fragmented DNA ranged from 6% to 12%. There was good correlation between the SCD and the sperm chromatin structure assay DNA fragmentation index (intraclass correlation coefficient R: 0.85; percent DNA fragmentation index mean difference: 2.16 significantly higher for SCD). Using the Halosperm kit, a dose-dependent increase in sperm DNA damage after sodium nitroprusside incubation was detected. The percentage of sperm cells with fragmented DNA in the fertile group was 16.3 +/- 6.0, in the normozoospermic group, 27.3 +/- 11.7, and in the oligoasthenoteratozoospermic group, 47.3 +/- 17.3. In the varicocele sample, an extremely high degree of nuclear disruption was detected in the population of sperm cells with fragmented DNA. CONCLUSION(S): The improved SCD test, developed as the Halosperm kit, is a simple, cost effective, rapid, reliable, and accurate procedure, for routinely assessing human sperm DNA fragmentation in the clinical andrology laboratory. PMID- 16213831 TI - Critically short telomeres are associated with sperm DNA fragmentation. AB - The influence of critical telomeric attrition, a well-known trigger of apoptosis and cell arrest, on sperm DNA fragmentation was studied in late-generation knockout mice for Terc, the RNA component of telomerase, as a model of choice. Terc knockout mice had a sixfold mean increase in the percentage of sperm cells with fragmented DNA. PMID- 16213832 TI - Comparison of the Halosperm test kit with the sperm chromatin structure assay (SCSA) infertility test in relation to patient diagnosis and prognosis. AB - The Halosperm test kit, the latest of the sperm DNA fragmentation tests, is considered by Fernandez et al. to be a suitable replacement for the Sperm Chromatin Structure Assay (SCSA) test. Although the test is ingenious and interesting, the data lack the statistical rigor of the SCSA test. PMID- 16213833 TI - Sperm DNA damage assessment: a test whose time has come. AB - Multiple techniques have been developed to measure the amount of sperm DNA damage in an effort to identify more objective parameters for evaluation of infertile men. We now have evidence to support that integrity of sperm DNA influences a couple's fertility and helps predict the chances of pregnancy and its successful outcome. The available tests of sperm DNA damage require additional large-scale clinical trials before their integration into routine clinical practice. PMID- 16213834 TI - Yet another test of sperm chromatin structure. AB - Sperm chromatin can affect reproductive performance, and it may be analyzed by measuring susceptibility of DNA to breakage using assays such as the sperm chromatin structure assay, comet assay, TUNEL, and DNA ladders. The newly proposed test, Halosperm, may not provide additional information beyond that obtained with existent evaluations. PMID- 16213835 TI - Halosperm is an easy, available, and cost-effective alternative for determining sperm DNA fragmentation. AB - The characteristics of Halosperm make this kit a reasonable alternative to allow basic and clinical research on sperm DNA fragmentation in any basic laboratory around the world. PMID- 16213836 TI - Harmonizing legal and ethical standards for interactions between health care providers and industry. AB - A complex set of federal and state laws, together with numerous voluntary codes, regulate the provision of gifts, consulting arrangements, and other interactions between industry and health care professionals. The provisions of these laws and codes are reviewed, and a suggested harmonization with respect to the most common types of interactions is proposed. PMID- 16213837 TI - Effect of coasting on the implantation potential of embryos transferred after cryopreservation and thawing. AB - OBJECTIVE: To examine the effect of withholding gonadotropins on the outcome of embryos after cryopreservation and thawing. DESIGN: Retrospective clinical evaluation of patients having cryopreserved-thawed ET trials with coasting during the corresponding ovarian stimulation cycle. SETTING: Academic tertiary clinical care unit. PATIENT(S): Patients with cryopreserved embryos having coasting in their fresh IVF cycle and age-matched controls without coasting, both groups receiving the same stimulation protocol (long GnRH agonist plus recombinant FSH). INTERVENTION(S): All patients had a cycle in which embryos were transferred fresh and a cycle of thawing of cryopreserved embryos with the aim of transferring in a steroid-supplemented cycle. MAIN OUTCOME MEASURE(S): Embryo survival, implantation, and clinical pregnancy rates. RESULT(S): Post-thawing embryo survival (66.4% vs. 73%), implantation (12.3% vs. 13.0%), and clinical pregnancy rates (31.5% vs. 38.0%) were similar in study and control groups, respectively. Patients with coasting for > or =3 days had significantly lower post-thawing embryo survival rates compared with patients having shorter duration of coasting (<3 days) and controls. Implantation and pregnancy rates, however, were similar in the three groups. CONCLUSION(S): Coasting did not seem to have a detrimental effect on oocyte and embryo quality because the implantation competence of transferred concept after cryopreservation and thawing was similar to that of controls. However, prolonged coasting (> or =3 days) had a subtle negative impact on the post-thaw survival rate. PMID- 16213838 TI - In vitro fertilization and embryo transfer in seminatural cycles for patients with ovarian aging. AB - OBJECTIVE: To investigate whether seminatural cycle is a reasonable management for ovarian aging patients. DESIGN: Prospective study. SETTING: ART Unit, Clamart, France. PATIENT(S): Seventy-five women, 158 cycles. INTERVENTION(S): Infertile women who presented with ovarian aging (defined as low ovarian reserve and characterized by cycle day 3 high FSH, high E2, and/or low inhibin B and/or previous cycle cancellations due to poor ovarian response to COH) were studied. Patients were offered up to three cycles. Treatment was scheduled as follows. From cycle day 8 onward the selection of the dominant follicle was monitored by ultrasound and hormonal measurements. When the dominant follicle appeared, patients received GnRH antagonist and, thereafter, hMG to support further follicular development. MAIN OUTCOME MEASURE(S): Implantation rate and clinical pregnancy. RESULT(S): Twenty-eight of 158 cycles were cancelled (17.7%). Oocyte pickups were performed in 119 (75.3%) cycles, 91 (57.6%) mature oocytes were retrieved, and 67 (42.4%) embryos transferred. Nineteen clinical pregnancies were obtained; the cumulative pregnancy rate per patient, after 3 cycles, was 35.2%. CONCLUSION(S): Use of a seminatural cycle is a reasonable management for patients with ovarian aging who have ovulatory menstrual cycles. It achieves a high implantation rate (28.3%). PMID- 16213839 TI - Early pronuclear breakdown is a good indicator of embryo quality and viability. AB - OBJECTIVE: To examine whether the timing of pronuclear breakdown can be a predictor of embryo quality and viability. DESIGN: Retrospective comparison of the development and quality of early and late developing zygotes. SETTING: Infertility and endocrinology unit in a university hospital. PATIENT(S): One thousand seven hundred eighty-two zygotes obtained in 383 consecutive IVF cycles. INTERVENTION(S): Culture of all fertilized embryos. MAIN OUTCOME MEASURE(S): Number of fertilized zygotes showing early pronuclear breakdown at 22-25 hours postinsemination, embryo quality, pregnancy rates (PR), implantation rates. RESULT(S): Early pronuclear breakdown embryos had a significantly higher cell number (4.4 +/- 1.2) compared with the late pronuclear breakdown embryos (3.6 +/- 1.4). When comparing the frequency of the early pronuclear breakdown embryos according to the method of fertilization, we failed to find any significant difference between the IVF (37.1%) and the intracytoplasmic sperm injection (ICSI) (41.1%) groups. The transfer of early pronuclear breakdown embryos resulted in a significantly higher clinical pregnancy rate than those with late pronuclear breakdown (48.3% vs. 27.3%). The implantation rate was higher in the early pronuclear breakdown group than in the late pronuclear breakdown group (26.5% vs. 15.1%). CONCLUSION(S): Early pronuclear breakdown is a strong indicator of embryo viability, and may be used as an additional criterion in the selection of embryos for transfer. PMID- 16213840 TI - Developmental potential of fully intact and partially damaged cryopreserved embryos after laser-assisted removal of necrotic blastomeres and post-thaw culture selection. AB - OBJECTIVE: Partially damaged frozen and thawed embryos are currently considered to have a lower viability than intact ones. This study was undertaken to compare the performance of intact frozen and thawed embryos with that of partially damaged embryos after removal of the necrotic blastomeres. DESIGN: Observational clinical series. SETTING: Private hospital. PATIENT(S): Three hundred twenty-six infertile couples undergoing frozen embryo transfer. INTERVENTION(S): Removal of necrotic blastomeres from frozen-thawed human embryos. MAIN OUTCOME MEASURE(S): Pregnancy and implantations rates. RESULT(S): Outcomes of frozen embryo transfer cycles in which all embryos were fully intact (group 1) were compared with those in which all embryos have lost 1-2 blastomeres (group 2) or 3-4 blastomeres (group 3). Laser-assisted hatching was performed in all embryos, and necrotic blastomeres were removed from partially damaged embryos on this occasion. Only embryos that resumed mitotic activity after thawing were transferred. Comparable clinical pregnancy rates (PR) (38.7%, 39.6%, and 29.4%), delivery rates (34.4%, 34.0%, and 29.4%), and implantation rates (21.6%, 21.4%, and 17.2%) were obtained in groups 1, 2, and 3, respectively. CONCLUSION(S): The developmental potential of partially damaged frozen and thawed embryos can be equivalent to fully survived embryos if the necrotic blastomeres are removed from the partially damaged embryos and only those of them that show post-thaw cleavage are selected for transfer. PMID- 16213841 TI - A bovine protocol for training professionals in preimplantation genetic diagnosis using polymerase chain reaction. AB - OBJECTIVE: To develop a bovine protocol for training in preimplantation genetic diagnosis (PGD) using PCR. DESIGN: Randomized study. SETTING: Human reproduction PCR laboratory. PATIENT(S): Cow ovaries obtained from slaughterhouses. INTERVENTION(S): The ovaries were punctured and the oocytes were matured and submitted to in vitro fertilization. On the third day after fertilization, the embryos were biopsied and 1-2 blastomeres removed. A blastomere and the rest of the embryo were submitted to PCR for sex determination. MAIN OUTCOME MEASURE(S): Establishment of a possible training protocol. RESULT(S): A total of 50 embryos and 50 biopsied blastomeres were submitted to DNA amplification for sexing. Of the 50 embryos, 41 (82%) achieved successful DNA amplification and 9 (18%) did not. Of the 50 biopsies, 31 (62%) amplified and 19 (38%) did not. In 27 (65.9%) of the 41 embryos with DNA amplification, sex was identified as female and in 14 (34.1%) as male. In 40 cases (80%) amplification and sex determination were successful in both embryos and blastomeres. Sex was identical in all these cases. CONCLUSION(S): This training model seems to be useful in identifying mistakes and difficulties and improving the professional's performance in the various stages of preimplantation genetic diagnosis. PMID- 16213842 TI - Relationship between pre-embryo pronuclear morphology (zygote score) and standard day 2 or 3 embryo morphology with regard to assisted reproductive technique outcomes. AB - OBJECTIVE: To test the hypothesis that pregnancy rates are low if grade Z1 pre embryos are not available for transfer and to determine if pronuclear morphology is a better predictor of pregnancy than traditional embryo morphology. DESIGN: Prospective clinical study. SETTING: Academic human reproduction laboratory. PATIENT(S): One hundred couples undergoing IVF with conventional insemination or ICSI. INTERVENTION(S): Embryo quality was assessed using both pre-embryo pronuclear morphology (zygote scoring or Z-scoring) at the time of fertilization evaluation and standard day 2 and day 3 embryo morphology (number of blastomeres and grading based on degree of fragmentation and blastomere size). MAIN OUTCOME MEASURE(S): We tested two decision models, one based on Z-scores and another on morphology, to determine which grading system better predicted pregnancy outcomes in assisted reproductive technique. Zygote score and embryo morphology were measured for all embryos and the transferred embryo pool. Implantation and pregnancy rates resulting from the embryo transfers of all cycles were calculated. RESULT(S): The Z-score distribution of 552 embryos was 27% Z1, 8% Z2, 50% Z3, and 15% Z4. Z1 and Z3 embryos had significantly (P approximately .03) higher quality over Z2 and Z4 embryos. Using the Z-score decision model with Z1 embryos having highest priority for transfer, pregnancy rates were similar between Z1 and Z3 embryos. Using embryo morphology as a decision model, pregnancy rates were highest in transfers containing one or two "best"-quality embryos. CONCLUSION(S): Z1 and Z3 embryos had similar morphology and pregnancy rates. The decision model based on the Z-score model was not better than standard embryo morphology in predicting pregnancy outcome. PMID- 16213843 TI - Multilocus analyses of estrogen-related genes reveal involvement of the ESR1 gene in male infertility and the polygenic nature of the pathology. AB - OBJECTIVE: To examine whether polymorphisms within the ESR1, FSHR, ESR2, CYP19A1, and NRIP1 genes are susceptibility factors for human male idiopathic infertility and to test the joint effects of these genes on male reproductive function. DESIGN: Genetic association study of male infertility with polymorphisms, using both single-gene and multilocus approaches. SETTING: Private and public fertility units and a private center for biomedical research. PATIENT(S): One hundred four Spanish men with azoospermia or severe oligozoospermia and 95 unselected race matched healthy controls from the same geographic region. INTERVENTION(S): Peripheral blood extraction, DNA purification, and ESR1 g.938T>C, FSHR Ser680Asn, ESR2 *39A>G, CYP19A1 *19C>T, and NRIP1 Gly75Gly polymorphism analyses. MAIN OUTCOME MEASURE(S): Single-gene statistical analyses and multilocus statistical analyses with Sumstat, Permutation and Model-free analysis, and Estimating Haplotypes software. RESULT(S): We observed an excess of homozygous infertile men for the ESR1 g.938T>C marker. Multilocus analyses detected genetic interaction between the five candidate gene markers that are influential over male infertility. In addition, we detected a five-loci protector genetic pattern with a frequency of 9.4% in controls but absent in infertile men. CONCLUSION(S): Our results support a relevant role for the estrogenic pathway, notably the ESR1 gene, in human male reproductive function and advocate a complex trait model for male infertility. PMID- 16213844 TI - Effect of chronic alcoholism on male fertility hormones and semen quality. AB - OBJECTIVE: To evaluate the effects of chronic alcoholism on the male fertility hormones and quality of semen. DESIGN: Non-probability purposive clinical study. SETTING: Addiction treatment center and an academic research environment. PATIENT(S): Sixty-six alcoholics free from smoking and drug abuse who consumed a minimum of 180 mL of alcohol per day (brandy and whisky, both 40%-50% alcohol content) for a minimum of 5 days per week for > or =1 year were included. Thirty nonsmoking nonalcoholics were selected as controls. INTERVENTION(S): Before starting the addiction treatment for alcoholics, venous blood and semen samples were collected. MAIN OUTCOME MEASURE(S): Complete blood counts, biochemical parameters, levels of the male fertility hormones FSH, LH, T, PRL, P, and E2 in blood, and semen parameters. RESULT(S): In alcoholics, FSH, LH, and E2 levels were significantly increased, and T and P levels were significantly decreased. No significant change was noted in PRL levels. Semen volume, sperm count, motility, and number of morphologically normal sperm were significantly decreased. CONCLUSION(S): Chronic alcohol consumption has a detrimental effect on male reproductive hormones and on semen quality. PMID- 16213845 TI - Comparison of the sperm aneuploidy rate in severe oligozoospermic and oligozoospermic men and its relation to intracytoplasmic sperm injection outcome. AB - OBJECTIVE: To determine the incidence of disomy and diploidy for chromosomes 18, X, and Y in the sperm samples of severe oligozoospermic (<5 x 10(6) spermatozoa/mL) and oligozoospermic (5-20 x 10(6) spermatozoa/mL) men undergoing intracytoplasmic sperm injection (ICSI) and to evaluate the influence of sperm aneuploidy on pregnancy outcome. DESIGN: Prospective study. SETTING: Infertility clinic and genetic laboratory. PATIENT(S): Fifteen patients with severe oligozoospermia, 15 patients with oligozoospermia, and 10 normal fertile donors. INTERVENTION(S): Fluorescence in-situ hybridization (FISH) performed on sperm samples. MAIN OUTCOME MEASURE(S): The frequency of disomy and diploidy for chromosomes 18, X, and Y was analyzed using FISH, and the clinical outcome after ICSI was correlated. RESULT(S): Significantly greater frequencies of XY, YY disomy and diploidy were observed in severe oligozoospermic men compared with oligozoospermic and normozoospermic men. Although the fertilization rate was similar, the pregnancy rate was higher in the group with oligozoospermia versus severe oligozoospermia. CONCLUSION(S): This study demonstrated the presence of an elevated sperm aneuploidy rate in patients with low semen quality. Additionally, the data show a negative influence of sperm chromosome abnormalities on ICSI outcome. PMID- 16213846 TI - Outcome of intracytoplasmic sperm injection in patients with polycystic ovary syndrome or isolated polycystic ovaries. AB - OBJECTIVE: To determine the intracytoplasmic sperm injection (ICSI) and embryo transfer (ET) performance of patients with polycystic ovary syndrome (PCOS) and isolated polycystic ovarian (PCO) morphology. DESIGN: Case-control study. SETTING: IVF Center, Hacettepe University Faculty of Medicine. PATIENT(S): Ninety nine consecutive infertile women (n = 109 cycles) with PCOS and 58 patients (n = 58 cycles) with isolated PCO morphology were recruited. The control group consisted of 210 patients (n = 232 cycles) with isolated male factor infertility necessitating ICSI. All three groups were matched for female age and body mass index. INTERVENTION(S): Controlled ovarian hyperstimulation and ICSI. MAIN OUTCOME MEASURE(S): Oocyte number, fertilization rate, embryo quality, clinical pregnancy rate, implantation rate, and ovarian hyperstimulation syndrome (OHSS). RESULT(S): Six (5.5%) cycles in the PCOS group, 6 (10.3%) cycles in the PCO-only group, and 10 cycles (4.3%) in the control group were canceled. Despite a significantly lower total FSH dose used, a significantly higher serum E2 level was attained in both the PCOS and the PCO-only groups compared to the control group. The PCOS and PCO-only groups had significantly higher numbers of retrieved oocyte-cumulus complexes and metaphase II oocytes compared to the control group. The fertilization rates did not differ among the three groups. The mean number of embryos transferred was comparable among the three groups; however, the mean number of grade 1 embryos was significantly higher in the PCOS and PCO-only groups compared to the controls. The clinical pregnancy rates per ET of both the PCOS (66%) and the PCO-only (60%) groups were significantly higher than that of the control group (44%). However, the implantation rates were comparable among the three groups. Four cycles (3.7%) in the PCOS group had OHSS necessitating hospitalization. The respective figures in the PCO-only and the control groups were 1 (1.7%) and 3 (1.3%). CONCLUSION(S): Patients with the full-blown picture of PCOS or isolated PCO-only morphology behave exactly in the same manner during all stages of assisted reproduction. Owing to the availability of more fertilized oocytes and grade 1 embryos, patients with PCOS or PCO-only morphology are associated with higher clinical pregnancy rates per ET compared to patients with isolated male factor infertility. PMID- 16213847 TI - Serum Mullerian-inhibiting substance levels in adolescent girls with normal menstrual cycles or with polycystic ovary syndrome. AB - OBJECTIVE: To compare serum mullerian-inhibiting substance (MIS) concentrations in adolescent girls with polycystic ovary syndrome (PCOS) or normal menstrual cycles. DESIGN: Prospective study. SETTING: University department of obstetrics and gynecology. PATIENT(S): Thirty-one girls (12-18 years old) with PCOS and 17 girls (12-19 years old) with normal menstrual cycles. INTERVENTION(S): Serum was collected from girls with PCOS or normal cycles during the early follicular phase of the menstrual cycle, stored frozen until assayed. MAIN OUTCOME MEASURE(S): Serum levels of MIS, E2, free-T, androstenedione, LH, and FSH. RESULT(S): Serum MIS levels in girls with PCOS were significantly higher compared with normal girls (4.1 +/- 2.2 [SD] and 2.4 +/- 1.0 ng/mL, respectively). The subjects were stratified for body mass index (BMI) (< and >25 kg/m2). Serum MIS levels in PCOS girls (4.2 +/- 3.0 ng/mL [BMI <25 kg/m2] and 4.0 +/- 1.6 [BMI >25 kg/m2]) were significantly higher compared with normal girls (2.2 +/- 0.8 and 2.6 +/- 0.7 ng/mL, respectively). CONCLUSION(S): Adolescent girls with PCOS have significantly higher serum MIS levels compared with normally cycling girls. Serum MIS levels in PCOS were not influenced by BMI. Increased MIS production may represent an early manifestation of the disease. PMID- 16213848 TI - Fertility after laparoscopic colorectal resection for endometriosis: preliminary results. AB - OBJECTIVE: To examine fertility, reproductive outcomes, and determinants of fertility after laparoscopic segmental colorectal resection for endometriosis. DESIGN: Retrospective longitudinal study. SETTING: Tertiary university gynecology unit. PATIENT(S): The study population consisted of 34 women with colorectal endometriosis, of whom 22 wished to conceive. Demographic, surgical, and histological characteristics of 10 women who conceived were compared with those of 12 women who failed to conceive. INTERVENTION(S): Laparoscopic colorectal resection for endometriosis. MAIN OUTCOME MEASURE(S): Rates of pregnancy and live birth. RESULT(S): Mean follow-up after segmental colorectal resection was 24 months (range 6-42 months), and the pregnancy rate was 45.5%. The median time to conceive was 8 months (range 3-13 months). Twelve pregnancies occurred in 10 women, comprising nine spontaneous singleton pregnancies (7 vaginal deliveries, 1 cesarean section, and 1 ongoing pregnancy), and three pregnancies obtained by IVF (one miscarriage, one ongoing twin pregnancy, and one triplet pregnancy necessitating cesarean section at 29 weeks for premature rupture of the membranes, with two surviving infants). The live birth rate was 82%. The women who did and did not conceive did not differ in terms of mean follow-up, mean age, body mass index (BMI), parity, smoking, use and duration of oral contraception (OC), duration of infertility, or the length of the resected colorectal segment. Uterine adenomyosis was the main determinant of pregnancy after colorectal resection. CONCLUSION(S): These preliminary results suggest that extensive laparoscopic segmental colorectal resection for endometriosis can enhance fertility, with high rates of spontaneous pregnancy and live birth. PMID- 16213849 TI - The prevalence of gonadotropin-releasing hormone receptor mutations in a large cohort of patients with hypogonadotropic hypogonadism. AB - OBJECTIVE: To determine the prevalence of GNRH receptor (GNRHR) gene mutations in a large cohort of patients with idiopathic hypogonadotropic hypogonadism (IHH). DESIGN: Molecular analysis and genotype/phenotype correlations. SETTING: University molecular reproductive endocrinology laboratory. PATIENT(S): North American and Turkish patients with IHH. INTERVENTION(S): DNA from 185 IHH patients were subjected to denaturing gradient gel electrophoresis for exons and splice junctions of the GNRHR gene. Variant fragments were sequenced. MAIN OUTCOME MEASURE(S): GNRHR mutations were characterized and compared with the phenotype. The prevalence of GNRHR mutations was also determined. RESULT(S): Three of 185 (1.6%; confidence interval [CI] 0.3%-4.7%) total IHH patients demonstrated compound heterozygous GNRHR mutations. All three were identified from a cohort of 85 normosmic patients (3.5%, CI 0.73%-7.5%), and none were demonstrated in hyposmic or anosmic IHH patients. GNRHR mutations were identified in 1 of 15 (6.7%; CI 0.2%-32.0%) families with at least two affected siblings, and in 2 of 18 (11.1%; CI 1.4%-34.7%) normosmic females. None were found in presumably autosomal dominant families. CONCLUSION(S): GNRHR mutations account for approximately 3.5% of all normosmic and 7%-11% of presumed autosomal recessive IHH, suggesting that additional genes play an important role in normal puberty. We believe this to be the largest GNRHR gene mutation analysis performed to date in a population of IHH patients. PMID- 16213850 TI - Autoimmune oophoritis as a mechanism of follicular dysfunction in women with 46,XX spontaneous premature ovarian failure. AB - OBJECTIVE: To assess the association between serum adrenal cortex autoantibodies and histologically confirmed autoimmune lymphocytic oophoritis. DESIGN: Controlled, prospective. SETTING: Tertiary research center. PATIENT(S): Two hundred sixty-six women with 46,XX spontaneous premature ovarian failure. INTERVENTION(S): Ovarian biopsy in 10 women. MAIN OUTCOME MEASURE(S): Serum adrenal cortex autoantibodies assessed by indirect immunofluorescence and autoimmune oophoritis assessed by immunohistochemical lymphocyte markers. RESULT(S): We obtained a histologic diagnosis of autoimmune oophoritis in four women who tested positive for adrenal cortex autoantibodies and excluded this diagnosis in ovarian biopsies from six women who tested negative for adrenal cortex autoantibodies (4/4 vs. 0/6). Women with histologically confirmed autoimmune oophoritis had a greater total ovarian volume as assessed by transvaginal sonography (11.4 +/- 5.6 mL vs. 1.5 +/- 0.4 mL) (mean +/- SEM). They were also more likely to have subclinical adrenal insufficiency and clinical signs of androgen deficiency (3/4 vs. 0/6). Overall, 10/266 women tested positive for adrenal cortex autoantibodies (3.8%, 95% confidence interval: 1.8%-6.5%). CONCLUSION(S): In women who present with 46,XX spontaneous premature ovarian failure as their primary concern there is a clear association between serum adrenal cortex autoantibodies and the presence of histologically confirmed autoimmune oophoritis. PMID- 16213851 TI - Time to pregnancy and pregnancy outcome. AB - OBJECTIVE: To investigate whether the outcome of a pregnancy is related to the time required to achieve that pregnancy (TTP). DESIGN: The distribution of the TTP for pregnancies ending in multiple birth, early (before week 12) and late (weeks 12-28) miscarriage, stillbirth, and extrauterine pregnancy was compared to that of pregnancies ending in singleton birth. Furthermore, the distribution of the TTP for preterm singleton births was compared to that of full-term singleton births. SETTING: Sweden. PATIENT(S): Information from three previous studies on reproduction was used: Women chosen for exposure to persistent organochlorine pollutants, or exposure as a hairdresser, and their respective controls. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Self-reported pregnancy outcome. RESULT(S): An increased TTP (i.e., decreased fecundability) was associated with pregnancies ending in miscarriage (early as well as late) and extrauterine pregnancies. Pregnancies ending in multiple live birth tended to have shorter TTPs than those ending in single live birth. No association between TTP and stillbirths was found. Among women whose pregnancies ended in singleton birth, a prolonged TTP was associated with preterm delivery. CONCLUSION(S): The TTP of a pregnancy seemed to be associated with the outcome of that pregnancy. The mechanisms behind this phenomenon are, however, unclear. PMID- 16213852 TI - The incidence of Sheehan's syndrome after obstetric hemorrhage. AB - OBJECTIVE: To estimate the incidence of Sheehan's syndrome in a well-described cohort of patients with obstetric hemorrhage. DESIGN: Retrospective cohort study. SETTING: Tertiary care center. PATIENT(S): Two hundred patients. INTERVENTION(S): Questionnaires were sent to study and comparison patients asking about menstrual dysfunction, lactation difficulty, cold intolerance, fatigue, axillary and pubic hair loss, and secondary infertility. MAIN OUTCOME MEASURE(S): Women who experienced two or more symptoms were referred for hormone testing of insulin like growth factor 1 (IGF-1), T4, PRL, and early morning cortisol (F) levels. RESULT(S): A total of 109 patients responded to the survey, a 55% response rate. Fourteen of 55 (25%) patients in the hemorrhage group identified themselves as suffering from two or more symptoms on the questionnaire. Eight of the 14 patients were tested, but none had hormonal evidence of hypopituitarism. Four of 54 (7%) comparison patients also identified themselves as suffering from two or more symptoms, but neither of the two tested had hormonal evidence of hypopituitarism. CONCLUSION(S): Among women with postpartum hemorrhage, subsequent development of clinical symptoms does not correlate well with laboratory evidence of hypopituitarism. Clinically significant Sheehan syndrome is an uncommon consequence of obstetric hemorrhage in today's environment. PMID- 16213853 TI - Prednisolone reduces preconceptual endometrial natural killer cells in women with recurrent miscarriage. AB - OBJECTIVE: To test the hypothesis that high numbers of uterine natural killer (uNK) cells in the endometrium of women with recurrent miscarriage (RM) could be reduced with prednisolone. DESIGN: A before and after study. SETTING: A tertiary referral teaching hospital. PATIENT(S): Eighty-five women with idiopathic RM recruited from all over the UK and 18 women attending for sterilization (controls). INTERVENTION(S): An endometrial sample was taken on day 21 +/- 2 of the menstrual cycle. Immunohistochemistry was used to identify uNK (CD56+, CD16-, CD3-). Twenty-nine women with RM and >5% uNK agreed to take 20 mg oral prednisolone daily from day 1 to 21 of their menstrual cycle, when a second biopsy was obtained and analyzed. MAIN OUTCOME MEASURE(S): The percentage of stromal cells that were uNK. The normal range was defined using control samples as <5%. RESULT(S): Women with RM had significantly more uNK than the controls (P=.008). Prednisolone treatment significantly reduced the number of CD56 cells in the endometrium, from a median of 14% (before) to 9% (after) (P=.0004). CONCLUSION(S): We have demonstrated that high numbers of uNK in preimplantation endometrium of women with recurrent miscarriage can be reduced with administration of prednisolone. PMID- 16213854 TI - Ovarian imaging by magnetic resonance in obese adolescent girls with polycystic ovary syndrome: a pilot study. AB - OBJECTIVE: To determine whether magnetic resonance (MR) imaging can serve as a useful investigational tool in the assessment of the polycystic ovary as compared with transabdominal ultrasound (US) for obese adolescents with polycystic ovary syndrome (PCOS). DESIGN: Prospective observational study. SETTING: Tertiary university hospital. PATIENT(S): Eleven obese adolescents with PCOS. INTERVENTION(S): Pelvic MR and US imaging and blood sampling. MAIN OUTCOME MEASURE(S): Total ovarian volume and follicle count; mean follicle count per longitudinal cross-section; stromal area; biochemical correlations with measured ovarian parameters. RESULT(S): With MR, the mean (+/-SE) total follicle count (21.9 +/- 1.3) was significantly greater than that observed with US (5.5 +/- 1.7) and significantly correlated with total ovarian volume. Two-dimensional cross sectional analysis of the ovary by MR revealed a significantly greater mean follicle count (12.1 +/- 0.8) compared with the results obtained by US (3.0 +/- 0.5). The stromal area (173.3 +/- 25.1 mm2) was approximately 35% of the total ovarian surface area. No correlations were observed between biochemical indices and measured ovarian parameters. CONCLUSION(S): In contrast to US, MR provides vastly greater delineation of the structural components of the ovary in obese girls with PCOS and thus can serve as an excellent investigational technique to assess the morphological transformation of the adolescent ovary. PMID- 16213855 TI - Modern embryo transfer catheters and pregnancy outcome: a prospective randomized trial. AB - OBJECTIVE: Embryo transfer (ET) is the final crucial step in IVF treatment. The type of catheter used can affect the pregnancy rate (PR). In this prospective, randomized trial we compared the clinical PR between the Wallace and the Cook K Jet embryo transfer catheters. DESIGN: Prospective, randomized clinical trial. SETTING: A National Health Service Assisted Reproduction Unit. PATIENT(S): One hundred fifty women undergoing a fresh ET. Age more than 40 years, a high basal FSH, a previous difficult ET, or more than six previous ETs were the exclusion criteria. INTERVENTION(S): Women undergoing a fresh ET were randomized at the time of ET to either the Cook K-Jet or Wallace embryo transfer catheter. The randomization was stratified according to age and the number of previous ETs. MAIN OUTCOME MEASURE(S): Clinical PR. RESULT(S): There was no significant difference in the clinical PR between the Wallace and the Cook catheters (22/75 [29.3%] and 23/75 [30.6%], relative risk [RR]: 0.96 [95% confidence interval 0.58 1.58]). CONCLUSION(S): There is no significant difference in the PRs achieved by modern, soft, double-lumen ET catheters. PMID- 16213857 TI - Vitrification of mouse pronuclear embryos after polar body biopsy without direct contact with liquid nitrogen. AB - OBJECTIVE: To evaluate the viability of vitrified mouse pronuclear embryos after polar body biopsy by cooling directly in liquid nitrogen in comparison with cooling in closed 0.5 mL straw (aseptic system). DESIGN: In vitro culture after vitrification. SETTING: Department of Gynecological Endocrinology and Reproductive Medicine, University of Bonn, Germany. PATIENT(S): Superovulated mice. INTERVENTION(S): Biopsied embryos were vitrified, warmed, and cultured in vitro. MAIN OUTCOME MEASURE(S): Development after warming. RESULT(S): Development rates up to expanded blastocyst stage after in vitro culture were 25% in group with "direct" vitrification and 23% in group with "straw in straw" vitrification. CONCLUSION(S): Cryopreservation of biopsied mouse pronuclear embryos in open pulled straws, which are placed inside a hermetically closed container, guarantees a complete isolation of embryos from liquid nitrogen and avoids potential contamination by pathogenic microorganisms. The combination of plunging this container into liquid nitrogen and rapid warming makes this process as efficient as conventional vitrification. PMID- 16213856 TI - Use of a novel washing method combining multiple density gradients and trypsin for removing human immunodeficiency virus-1 and hepatitis C virus from semen. AB - OBJECTIVE: To determine the effectiveness of a novel treatment designed to remove human immunodeficiency virus (HIV) -1 and hepatitis C virus (HCV) from spiked semen and to evaluate sperm motility and viability after treatment. DESIGN: A prospective clinical laboratory-based study. SETTING: The human studies were conducted in academic and national research environments. The bovine study was conducted in an accredited research facility. PATIENT(S): Healthy volunteers provided the semen samples used in the human studies; abattoir-derived material was used for the bovine embryo production study. INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): Cytopathic, reverse transcriptase-polymerase chain reaction, and branched DNA assays were used to test the efficacy of the procedure for inactivating or removing viruses from spiked semen; standard semen evaluation criteria were used to assess the effects of the procedures on sperm motility and viability. RESULT(S): Trypsin exposure significantly reduced the infectivity of HIV-1. The triple density gradient treatment, with or without trypsin, had no detrimental affect on fresh or cryopreserved/thawed sperm 2-48 hours after treatment. The treatment of semen spiked with HIV-1 or HCV indicated that the procedure was effective for reducing viral copies to undetectable levels or below levels of clinical relevance. CONCLUSION(S): The procedure was effective for significantly inactivating or reducing HIV-1 and HCV in spiked semen without adversely affecting sperm quality. PMID- 16213858 TI - Pseudomyxoma peritonei: a rare differential diagnosis of hydrosalpinx. AB - OBJECTIVE: To establish the diagnosis of a right hydrosalpinx in a woman with primary infertility. DESIGN: Case report. SETTING: Departments of Obstetrics and Gynecology and Surgery, academic medical center and teaching hospital. PATIENT(S): A healthy 41-year-old woman with a history of primary infertility and a possible right hydrosalpinx. INTERVENTION(S): Operative laparoscopy, disconnection of right hydrosalpinx, peritoneal biopsies. MAIN OUTCOME MEASURE(S): Establishment of diagnosis with peritoneal biopsies. RESULT(S): The patient was diagnosed with peritoneal pseudomyxoma peritonei, which required further treatment. CONCLUSION(S): Malignancies, although rare, should always be part of the differential diagnosis of external causes of tubal disease. PMID- 16213859 TI - Successful sperm storage for 28 years. AB - OBJECTIVE: To report on two instances of successful long-term cryopreservation resulting in live births. DESIGN: Case report. SETTING: The patient stored his sperm at a private sperm banking facility, and his partner underwent artificial insemination at an obstetrics/gynecology practice for one pregnancy and at a fertility center for the other. PATIENT(S): One man who stored his sperm before treatment for cancer. INTERVENTION(S): Storage of sperm under liquid nitrogen in a carousel canister system. Intrauterine insemination in which the semen was thawed, washed with human tubal fluid (HTF), and inseminated. MAIN OUTCOME MEASURE(S): Successful pregnancy resulting in live birth. RESULT(S): Artificial insemination with semen cryopreserved for 21 and 28 years resulted in two live births. CONCLUSION(S): This case report describes the to-date longest known successful cryopreservation of sperm, with two live births resulting from IUI. Successful long-term semen storage can be very beneficial for men facing impaired fertility or sterility early in life, so that they have sufficient time to make appropriate family planning decisions. PMID- 16213860 TI - Successful pregnancy and delivery after transfer of a single blastocyst derived from a vitrified mature human oocyte. AB - OBJECTIVE: To describe the first case of delivery after the transfer of a single blastocyst derived from a vitrified mature human oocyte. DESIGN: Case report. SETTING: Private assisted reproduction clinic. PATIENT(S): A normal 31-year-old woman. INTERVENTION(S): An unsuccessful attempt was made to extract sperm from the patient's nonobstructive azoospermic husband by testicular sperm extraction. MAIN OUTCOME MEASURE(S): Transfer of single blastocyst derived from vitrified human oocyte and donor sperm. RESULT(S): A healthy male neonate weighing 3000 g was born. CONCLUSION(S): Vitrification is a useful method of preserving mature human oocytes and has the advantage of time-effectiveness, simplicity, cost reduction, and no need for devices such as programmable freezers. PMID- 16213861 TI - Accessory uterine appendage may be a new Mullerian malformation. AB - OBJECTIVE: To describe a case of an accessory uterine structure. DESIGN: Case report. SETTING: University affiliated, community-based hospital. PATIENT(S): A 35-year-old woman who presented for laparoscopic tubal sterilization. INTERVENTION(S): Laparoscopy, tubal sterilization, dye test, and excision of accessory uterine structure. MAIN OUTCOME MEASURE(S): Laparoscopic findings and resection of the anomaly. RESULT(S): Laparoscopy showed a hollow uterine appendage that communicated with the uterine cavity. This was completely excised. CONCLUSION(S): We report a rare case of a uterine anomaly that is inconsistent with the traditional classification system, and we offer a possible mechanism for its formation. PMID- 16213862 TI - Orthotopic transplantation of fresh ovarian cortex: a report of two cases. AB - OBJECTIVE: To report two cases of orthotopic transplantation of fresh ovarian tissue. SETTING: Academic hospital. PATIENT(S): Two patients with severe endometriosis, who underwent left oophorectomy for recurrent endometriosis. INTERVENTION(S): Ovarian cortex was reimplanted in the heterolateral orthotopic site. RESULT(S): Biopsies of the grafted tissue were taken 3 months after reimplantation. Viable primordial follicles were found. The presence of a neovascular capillary network was demonstrated. CONCLUSION(S): Reimplantation of fresh ovarian cortex allows the survival of primordial follicles and may represent an alternative method for the preservation of ovarian cortex when oophorectomy is mandatory. PMID- 16213863 TI - SYCP3 mutations are uncommon in patients with azoospermia. AB - Mutation analysis of the SYCP3 gene was performed for 58 patients with a maturation arrest of spermatogenesis. In contrast to the previously reported high frequency of mutations, only polymorphisms were found in the present study. PMID- 16213864 TI - High-density lipoprotein particles may regulate hemostasis in human pregnancy. AB - We explored the association of apolipoprotein A-I (the major protein constituent of high-density lipoprotein (HDL)) and fibrin fragment D-dimer (a marker of cross linked fibrin turnover) in pregnant women. A significant inverse correlation between the levels of apolipoprotein A-I and fibrin fragment D-dimer was observed in both pre-eclamptic and normal pregnant women (rs = -0.459, n = 37 and rs = 0.290, n = 55, respectively), suggesting a major regulating role of HDL in hemostasis during pregnancy. PMID- 16213865 TI - Use of a luteal estradiol patch and a gonadotropin-releasing hormone antagonist suppression protocol before gonadotropin stimulation for in vitro fertilization in poor responders. AB - The administration of a luteal E2 patch/GnRH antagonist protocol before gonadotropins in poor responders may improve ovarian stimulation and result in greater uniformity in follicular development and improved pregnancy rates. PMID- 16213866 TI - In vitro oocyte maturation and subsequent delayed fertilization is associated with increased embryo aneuploidy. AB - Failed fertilization and the appearance of immature oocytes are common in IVF practice; rescue intracytoplasmic sperm injection can be used as a therapy. However, this study indicates that embryos created after in vitro maturation and delayed intracytoplasmic sperm injection contain an increase in aneuploidy (79.7%) over control embryos (60.5%). Therefore, patients should be informed of the possible risk when presented with delayed intracytoplasmic sperm injection. PMID- 16213867 TI - Influence of clinical and ultrasound factors on the efficacy of misoprostol in first trimester pregnancy failure. AB - An observational study including 276 patients with early pregnancy failure was performed to evaluate the clinical and ultrasound factors influencing the efficacy of misoprostol in the treatment of first trimester pregnancy failure. Gestational age did not influence the efficacy of this treatment and the success rate was inversely proportional to parity. PMID- 16213868 TI - Role of the vaginally administered aromatase inhibitor anastrozole in women with rectovaginal endometriosis: a pilot study. AB - In the present nonrandomized pilot study we determined the role of the vaginally administered aromatase inhibitor anastrozole (0.25 mg anastrozole/d for 6 months) in the treatment of women with histologically proven rectovaginal endometriosis. In a series of 10 patients, dysmenorrhea, physical and social functioning, but not chronic pelvic pain and dyspareunia, improved during therapy. PMID- 16213869 TI - Effect of oral conjugated equine estrogen combined with medroxyprogesterone acetate on plasma homocysteine levels in postmenopausal women. AB - Conjugated equine estrogen alone or combined with medroxyprogesterone acetate lowered homocysteine levels in postmenopausal women. Regardless of the dosage of progestin used, there was no impact on homocysteine metabolism after 3 years of therapy. PMID- 16213870 TI - Doubling time of urine human chorionic gonadotropin after assisted reproductive technology. AB - In the present study, we measured urinary hCG after assisted reproductive technology to accurately calculate normal doubling time (DT) of hCG, and we compared DT values with prognosis of pregnancy. We clarified the correlation between normal DT values, calculated from urinary hCG levels, and viable and nonviable pregnancies. PMID- 16213871 TI - Pregnancy outcome in in vitro fertilization decreases to a plateau with repeated cycles. AB - Clinical pregnancy rate (CPR) and implantation rate (IR) in 1,177 patients who had 1,788 fresh, nondonor, nonPGD IVF cycles were highest in cycle 1, significantly declined in cycle 2, and reached a plateau for cycles 3-5 at a rate lower than in cycle 2. In patients >38 years of age CPR and IR in cycles 1 and 2 were significantly lower than in younger patients, but there was no decline in CPR or IR with advancing IVF attempts. PMID- 16213872 TI - Outcome of in vitro fertilization after transabdominal ultrasound-assisted embryo transfer with a full or empty bladder. AB - The use of ultrasound guidance has proven to be a key factor in performing embryo transfer in a gentle and atraumatic manner. However, despite the lower incidence of brusque maneuvers, bladder distension has not shown any positive impact on the IVF success rate. PMID- 16213873 TI - Difference in expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 in patients with persistent ovarian cysts. AB - Matrix metalloproteinase-1 (MMP-1), MMP-2, and MMP-9 expression with real-time quantitative polymerase chain reaction were analyzed in endometriotic and nonendometriotic ovarian cysts. Although MMP-1 was not detected, MMP-9 and MMP-2 were expressed in all of the cysts. In particular, in five of six nonendometriotic cysts (83.3%) MMP-2 expression was higher than in endometriotic cysts. These data may represent new molecular elements helpful in differential diagnosis of endometriotic lesions. PMID- 16213874 TI - Maternal C-reactive protein levels in patients undergoing frozen embryo replacement cycles: a prospective study. AB - This prospective study shows that the early pregnancy inflammatory response following frozen embryo replacement (FER) cycles may be absent or suppressed, in contrast to that following IVF. To our knowledge, this is the first study to evaluate the maternal inflammatory response following FER cycles, but larger studies are still required to explore this novel finding, and investigate whether this may explain the lower ongoing pregnancy rates generally achieved following FER cycles. PMID- 16213875 TI - Refining embryo transfer. PMID- 16213877 TI - Optimal study design and the CONSORT guidelines. PMID- 16213879 TI - Important features of study design. PMID- 16213880 TI - Measure of diagnostic accuracy for hysterography? PMID- 16213883 TI - Measure of diagnostic accuracy for hysterography? PMID- 16213884 TI - Surgical application of cDNA microarray technique. PMID- 16213886 TI - Hyperbolic hypotheses. PMID- 16213887 TI - The surgeon's use of evidence to care for our patients. PMID- 16213888 TI - Re: flaws in methods of evidence-based medicine may adversely affect public health directives. PMID- 16213889 TI - Re: flaws in methods of evidence-based medicine may adversely affect public health directives. PMID- 16213893 TI - Ligand-dependent activation of the epidermal growth factor receptor by secondary bile acids in polarizing colon cancer cells. AB - BACKGROUND: Secondary bile acids such as deoxycholic acid (DCA) are known to promote colorectal cancer (CRC). Increasing evidence suggests that DCA-induced signaling is mediated by activation of the epidermal growth factor receptor (EGFR). We have shown that activation of the EGFR induces up-regulation of cyclooxygenase 2, basolateral release of prostaglandins (PGs), and mitogenesis in a polarizing human colon cancer cell line, HCA-7. The purpose of this study was to determine the mechanism by which DCA activates EGFR in human polarizing CRC cell lines HCA-7 and HCT-8. METHODS: A primary, non-tumor-promoting bile acid (cholic acid [CA]) and a secondary, tumor-promoting bile acid, DCA, were added to the apical and basolateral compartment of polarized HCA-7 and HCT-8 cells. These cells were pretreated with monoclonal antibody 528, a monoclonal antibody that inhibits ligand binding to EGFR, or with WAY-022, a selective inhibitor of tumor necrosis factor-alpha converting enzyme/a disintegrin and metalloprotease-17 (TACE/ADAM-17), which cleaves amphiregulin (AR) to its mature, soluble form from the basolateral cell membrane. AR levels were measured in the apical and basolateral medium and cell lysates by radioimmunoassay. PGs were measured in the apical and basolateral medium by gas chromatography/mass spectrometry. RESULTS: Basolateral delivery of DCA, but not CA, preferentially stimulated release of AR into the basolateral medium compared with cell lysates of polarized HCA-7 and HCT 8 cells. Basolateral delivery of DCA resulted in increased basolateral PGE2 levels (P < .05), and this effect was attenuated by pretreatment with monoclonal antibody 528 (P < .05). Inhibiting cell surface cleavage of AR with WAY-022 before DCA treatment reduced AR (P < .05) and PGE2 (P < .05) levels in the basolateral medium. CONCLUSION: DCA, but not CA, results in compartment-specific, ligand-dependent activation of EGFR and subsequent increased basolateral PGE2 levels. The mechanism of DCA-induced EGFR activation is ligand-dependent and is controlled, at least in part, at the level of AR release from the basolateral cell membrane. PMID- 16213894 TI - Synergistic tumoricidal effect between celecoxib and adenoviral-mediated delivery of mda-7 in human breast cancer cells. AB - BACKGROUND: Celecoxib, a selective cyclooxygenase 2 (COX-2) inhibitor, blocks growth and promotes apoptosis in breast cancer cells. The PI3K/Akt pathway is important in cell survival, and COX-2 and Akt might promote growth via a positive feedback loop. We have shown that adenoviral delivery of mda-7 (Ad-mda7) in breast cancer down-regulates Akt. We hypothesized that combining Ad-mda7 and celecoxib could mediate tumor suppression in COX-2 overexpressing breast cancer cells. METHODS: Two COX-2 overexpressing human breast cancer cell lines (Her-18 and MDA-MB-436) were treated with celecoxib (20 micromol/L and 50 micromol/L) and Ad-mda7 (multiplicity of infection, 1000 and 2000 viral particles/cell). Adenovirus encoding the luciferase gene was used as a control. We assessed proliferation, cell cycle, apoptosis, prostaglandin E2 production, and changes in protein expression. Statistical analysis was performed by using the Student t test. RESULTS: Regardless of HER-2/neu status, cell growth was markedly inhibited by celecoxib, Ad-mda7, and the combination compared with controls. Celecoxib + Ad mda7 showed a greater than additive increase in cell death compared with either monotherapy (P < .05) and resulted in cell cycle block and apoptosis (P < .05). Both cell lines showed decreased prostaglandin E2 production after combination treatment compared with controls (P < .05), with decreased expression of COX-2, Akt, and phosphorylated Akt (P < .05). CONCLUSIONS: Enhanced antitumor activity is achieved in breast cancer by combining celecoxib and Ad-mda7 regardless of HER 2/neu status. This occurs through inhibition of COX-2 expression and down regulation of Akt. Combining Ad-mda7 with COX-2 inhibition provides a novel method of treatment in breast cancer. PMID- 16213895 TI - Ubiquitin reduces fluid shifts after traumatic brain injury. AB - BACKGROUND: Ubiquitin has well-described intracellular properties. Recent data also suggest pleiotropic effects of extracellular ubiquitin, including induction of apoptosis, regulation of immune functions, and therapeutic potential during fluid resuscitation from severe trauma. However, the actions of exogenous ubiquitin after traumatic brain injury (TBI) are unknown. METHODS: Series 1: Thirty-five minutes after TBI and hemorrhage, 1.5 mg ubiquitin/kg (n = 5) or albumin (n = 5) intravenous was followed by fluid resuscitation to maintain mean arterial and cerebral perfusion pressure. Series 2: Ubiquitin (n = 5) or vehicle (n = 6) was administered after TBI only. Ubiquitin was measured with enzyme linked immunosorbent assay in serum, urine (series 1), and cerebrospinal fluid (series 2) for 300 minutes. RESULTS: Series 1: After intravenous bolus, serum ubiquitin peaked at t = 45 minutes with a half-life of 54 minutes. Recovery in urine was 10%. With albumin versus ubiquitin, 85% more resuscitation fluid was required to stabilize systemic and cerebral hemodynamics (P < .05 for t = 150 to 300 minutes), but hematocrit was similar. With albumin there were progressive increases in intracranial pressure, peak inspiratory pressure, and decreases in oxygenation. All were significantly attenuated by ubiquitin (all P < .05 vs albumin). Series 2: Intravenous ubiquitin altered cerebrospinal fluid ubiquitin with an increased time to peak (t = 88 +/- 13 min vs 45 +/- 7 min, P < .05) and area under the concentration-time curve (82 +/- 22 vs 23 +/- 11 microg/min(1)/mL( 1), P < .05). CONCLUSIONS: After TBI, intravenous ubiquitin crossed the blood brain barrier and significantly reduced third spacing of fluid into the brain and lung during resuscitation. PMID- 16213896 TI - Normal human fibroblasts enable melanoma cells to induce angiogenesis in type I collagen. AB - BACKGROUND: We previously reported that fibroblasts induce human microvascular endothelial cells (HMVECs) to differentiate from monolayer to capillarylike morphology. We now test the hypothesis that fibroblasts modulate angiogenesis in melanoma cells. METHODS: We tested 12 human melanoma lines (2 radial growth phase (RGP), 3 vertical growth phase (VGP), and 7 metastatic (MM)) for ability to induce HMVECs to invade/migrate into collagen and form capillarylike networks. HMVEC monolayers were overlaid with 3-dimensional collagen gels embedded with melanoma cells alone (M), fibroblasts alone (F), or a 1:1 mixture of the 2 cells (M+F). After 5 days, gels were removed, fixed, and HMVEC networks were quantified by von Willebrand's factor (vWF) immunofluorescence. The influence of soluble factors on HMVEC invasion/migration into collagen was assessed with the use of acellular 3-D collagen gels overlaid on HMVEC monolayers, cultured with conditioned media (CM) derived from monolayers of M, F, or M+F. Angiogenic growth factors involved in the observed invasion/migration were identified with the use of a RayBio Cytokine Antibody Array (RayBiotech, Norcross, Ga). RESULTS: Cell line-specific variability in melanoma-supported angiogenesis was observed only when in combination with fibroblasts (analysis of variance [ANOVA], P < .01). Melanoma plus fibroblasts uniformly resulted in a significantly higher angiogenic response than melanoma alone (P < .05). One vertical growth phase and one metastatic melanoma line, while weakly angiogenic alone, induced significantly higher angiogenesis than either fibroblast or melanoma alone (P < .05) when combined with fibroblasts. CM from M or M+F induced significantly less HMVEC invasion/migration into collagen than CM from fibroblasts alone. Interleukin 8, monocyte chemotactic protein-1, and tissue inhibitor of metalloproteinase-2 were identified as significantly elevated in the media derived from M+F cultures, compared with either cell type alone. CONCLUSION: To our knowledge, this is the first report demonstrating that melanoma-supported angiogenesis in collagen is more significantly influenced by normal skin-derived fibroblasts than by the intrinsic biology of the melanoma cell type. Interleukin 8, monocyte chemotactic protein-1, and tissue inhibitor of metalloproteinase-2 are implicated as potential paracrine factors regulating this observed effect. PMID- 16213898 TI - Expression and prognostic value of tuberous sclerosis complex 2 gene product tuberin in human pancreatic cancer. AB - BACKGROUND: Tuberous sclerosis complex (TSC) is an autosomal dominant disorder caused by mutation of either of 2 tumor suppressor genes, TSC1 or TSC2, which encode hamartin and tuberin, respectively. Several studies have shown that tuberin functions independently of hamartin and inhibits signaling pathways via the mammalian target of rapamycin, a critical regulator of cell proliferation. Recent studies have revealed that the signaling pathways regulating the mammalian target of rapamycin such as Akt and S6K1 are frequently activated in pancreatic cancer. We hypothesized that tuberin might be involved in the proliferation and survival of pancreatic cancer cells. METHODS: We immunohistochemically examined the expression of tuberin in 42 pancreatic cancerous and noncancerous pancreatic tissue specimens using an antituberin antibody. The correlations between tuberin expression and various clinicopathologic features, including survival, were evaluated. Reverse transcriptase-polymerase chain reaction was performed to evaluate the level of tuberin expression in paired samples of pancreatic cancer and noncancerous tissue. RESULTS: Twenty-four of the 42 pancreatic cancer samples (57%) were negative for tuberin expression. The patients with tuberin-negative tumors had a significantly higher incidence of pT3 or pT4 disease (primary tumor extent by the TNM classification) than those with tuberin-positive tumors (P = .024). Female patients had a significantly higher incidence of tuberin-positive tumors than male patients (P = .014). The survival rate of the tuberin-positive group tended to be better than that of the tuberin-negative group, but there was no significant difference (P = .4). Expression of TSC2 in cancer tissue was lower than in the corresponding noncancerous tissue for 7 of the 9 samples examined. CONCLUSIONS: This study demonstrates that reduced expression of tuberin might be involved in the progression of pancreatic cancer. Accordingly, tuberin may provide a new therapeutic target in patients with this type of cancer. PMID- 16213899 TI - Modulation of parathyroid hormone-related protein levels (PTHrP) in anaplastic thyroid cancer. AB - BACKGROUND: Studies have demonstrated that manumycin, a farnesyltransferase inhibitor, enhances the cytotoxic effect of paclitaxel in anaplastic thyroid cancer cells and in xenografts, but the mechanism of this effect is unknown. Parathyroid hormone-related protein (PTHrP) may function as an oncoprotein that inhibits apoptosis and enhances cell proliferation, in addition to its role as the mediator of humoral hypercalcemia of malignancy. We hypothesized that this protein might have a novel role in anaplastic thyroid cancer. METHODS: Five anaplastic thyroid cancer cell lines (ARO, DRO, KAT-4, Hth-74, C-643) were examined for PTHrP expression in vitro by immunohistochemistry (IHC), radioimmunoassay, and Western blot (IP/WB) analyses. PTHrP expression was also examined in an in vivo xenograft model. The effects of manumycin and paclitaxel on PTHrP expression were studied. RESULTS: All 5 ATC cell lines were found to robustly express PTHrP by IHC of fixed cells and radioimmunoassay of cell lysates and conditioned culture media (range, 468 +/- 55 to 1410 +/- 195 pg/mg cellular protein). Manumycin (54 micromol/L), but not paclitaxel (22 micromol/L), decreased the amount of PTHrP. Further, PTHrP was decreased in KAT-4 xenografts in nude mice that had been treated for 3 weeks with biweekly intraperitoneal injections of manumycin (7.5 mg/kg), compared with control mice by IHC. On Western blot analyses, fractionation of radiolabeled proteins showed that manumycin decreased synthesis of PTHrP in cytoplasm, with the amount of newly synthesized PTHrP in the nucleus and increased ubiquitination of PTHrP suggesting increased degradation of PTHrP through the proteasome pathway. CONCLUSIONS: Manumycin inhibits cell proliferation and decreases PTHrP levels in anaplastic thyroid cancer cells in vitro and in vivo and decreases the PTHrP level in the nucleus where PTHrP may function as an oncoprotein. These data suggest that PTHrP has a novel role in anaplastic thyroid cancer and that modulation of PTHrP levels may be of therapeutic benefit in this lethal malignancy. PMID- 16213900 TI - Effects of lysophosphatidylcholine on monolayer cell permeability of human coronary artery endothelial cells. AB - BACKGROUND: Lysophosphatidylcholine (LysoPC) is a product of phosphatidylcholine hydrolysis by phospholipase A2, which is associated with atherosclerosis. However, the underlying molecular mechanisms are still unclear. The purpose of this study was to determine the effects of LysoPC on monolayer permeability of human coronary artery endothelial cells (HCAECs). METHODS: HCAECs were cultured with LysoPC in a dose- and time-dependent manner. Monolayer permeability was studied by using a transwell system with a Texas-Red-labeled dextran tracer. The messenger RNA and protein levels of endothelial tight junction proteins were determined with the use of real-time reverse transcriptase-polymerase chain reaction and Western blot analysis, respectively. Superoxide anion levels were determined with the use of fluorescent dye dihydroethidium-based flow cytometry analysis. Activation of mitogen-activated protein kinases was determined by performing Bio-Plex immunoassay. RESULTS: LysoPC (30 micromol/L) increased monolayer permeability by 53% and decreased the messenger RNA levels of zonula occludens-1, occludin, claudin-1, and junctional adhesion molecule by 44%, 53%, 50%, and 52%, respectively, compared with controls (P < .05). Western blot analysis showed reduced protein levels of these tight junction molecules. LysoPC (15 and 30 micromol/L) also increased superoxide anion production by 54% and 58%, respectively, compared with controls (P < .05). Antioxidant seleno-L-methionine (20 and 30 micromol/L) inhibited LysoPC (30 micromol/L)-induced permeability by 42% and 68%, respectively (P < .05). Furthermore, LysoPC (30 micromol/L) activated c-Jun N-terminal kinase and p38 phosphorylation, but not extracellular signal-related kinase 1/2, within 5 to 10 minutes. CONCLUSIONS: LysoPC increases monolayer permeability and reduces the expression of tight junction molecules in HCAECs through oxidative stress and activation of c-Jun N-terminal kinase and p38 mitogen-activated protein kinase. The antioxidant can effectively block LysoPC induced endothelial permeability. PMID- 16213901 TI - Molecular analysis improves sensitivity of breast sentinel lymph node biopsy: results of a multi-institutional prospective cohort study. AB - BACKGROUND: Pathologic evaluation may lack the sensitivity required for accurate staging of the axilla in breast cancer patients. We have completed enrollment of a multi-institutional prospective cohort study designed to determine if molecular analyses can improve axillary staging. In subset analyses, we have attempted to address the following questions: (1) Does molecular analysis improve the sensitivity of sentinel lymph node biopsy (SLNB) and (2) is the sentinel lymph node (SLN) hypothesis valid at the molecular level? METHODS: Four hundred eighty nine subjects with T1, T2, or T3 breast cancer and no evidence of axillary lymph node (ALN) involvement were enrolled. ALNs were analyzed by routine pathology (hematoxylin-eosin staining), and by multimarker real-time reverse transcriptase polymerase chain reaction analysis CRT-PCR to detect breast cancer metastases. Pathology and molecular data for both SLNs and nonsentinel ALNs were available for a subset of 207 subjects. RESULTS: The sensitivity of pathologic analysis of the SLN to predict the pathologic status of ALNs was 84.1%. The sensitivity of combining pathologic with molecular analysis of the SLN to predict the pathologic status of ALNs was 92.8%, a statistically significant increase in sensitivity (P = .031 by the McNemar test for correlated proportions). Finally, the sensitivity of combining pathologic with molecular analysis of the SLN to predict the pathologic or molecular status of ALNs was 85.4%. CONCLUSIONS: The combination of pathologic and molecular analysis of SLNs resulted in the highest sensitivity for prediction of the pathologic status of ALNs. The data also provide evidence that the SLN hypothesis remains valid at the molecular level. PMID- 16213902 TI - The effect of hepatocyte growth factor on gut mucosal apoptosis and proliferation, and cellular mediators after severe trauma. AB - BACKGROUND: A severe burn injury is associated with an impairment of gut mucosal integrity and function, which is due to increases in small-bowel epithelial cell apoptosis and decreases in cell proliferation. Hepatocyte growth factor (HGF) was shown to improve regeneration in the liver, mesentery, and skin. The purpose of this study was to determine whether HGF can improve small-bowel homeostasis after injury and the cellular mechanisms by which these changes occur. METHODS: Rats were pair-fed, underwent thermal trauma, and received saline (0.9% NaCl; n = 28) or HGF (200 microg/kg iv every 12 hours, n = 28). Small intestine and serum were taken at 1, 2, 5, and 7 days after injury. Measures were mucosal apoptosis, proliferation, villous morphology, and apoptotic and proliferative mediators, such as caspase-3 and caspase-7, Fas and Fas-ligand, Bcl-2, and Bax. In addition, serum cytokines were determined. RESULTS: Gut epithelial cell apoptosis was increased in the saline and HGF groups after the thermal injury. Despite an increase in serum tumor necrosis factor-alpha and interleukin-1beta, HGF did not affect small-bowel cell apoptosis, but it improved proliferation at days 1 and 2 after injury, which was associated with increased villous height and cell per villous, compared with saline controls, P < .05. Increased mucosal cell proliferation was associated with increased Bcl-2 in the HGF group, P < .05. HGF had no effect on apoptotic mediators, such as Fas, Fas-L, or caspase-3 and caspase-7. CONCLUSIONS: HGF improves small-bowel morphology after a severe burn by increasing mucosal Bcl-2 and, concomitantly, small-bowel epithelial cell proliferation. PMID- 16213903 TI - Six core competencies and seven deadly sins: a virtues-based approach to the new guidelines for graduate medical education. AB - As part of efforts afoot to improve the overall quality of physician training, the Accreditation Council for Graduate Medical Education (ACGME) has endorsed a set of competencies that will be the blueprint for outcomes-based graduate medical education for years to come. While the spirit of this new law is taking shape, the letter remains largely unwritten. To bridge this gap, administrators of programs from all specialties must determine how the core competencies will be taught, evaluated, modeled, and enforced within their respective programs. This paper summarizes these challenges, in particular for surgical programs, and focuses constructively on the modeling/enforcement approach, describing key characteristics that programs should pursue and cultivate (virtues) as well as the signal prohibitions (vices) that both trainees and trainers must avoid. Regardless of specialty or programmatic particulars, virtues and vices may be used to define a context in which general competencies may be understood, and yield operational guidance upon which ultimate discussions of evaluation, remediation, and graduation may be predicated. PMID- 16213904 TI - Present-at-admission diagnoses improve mortality risk adjustment and allow more accurate assessment of the relationship between volume of lung cancer operations and mortality risk. AB - BACKGROUND: Mortality risk adjustment is a key component of studies that examine the statistical relationship between hospital lung cancer operation volume and in hospital mortality. Previous studies of this relationship have used different methods of adjusting for factors that influence mortality risk, but none have adjusted for differences in comorbid disease using only diagnoses identified as present-at-admission. METHODS: This study uses adjustments for conditions identified as present-at-admission to examine the statistical relationship between the volume of lung cancer operations and mortality among 14,456 California hospital patients, and compares these results to other methods of risk adjustment similar to those used in previous studies. RESULTS: Mortality risk adjustment using present-at-admission diagnoses yielded better discrimination and explained more of the variability in observed deaths. Large increases in hospital procedure volume were associated with much smaller decreases in mortality risk than those estimated using comparable risk-adjustment models. CONCLUSIONS: Present-at-admission diagnoses can be used to improve mortality risk adjustment and may allow a more accurate assessment of the relationship between procedure volume and mortality risk. PMID- 16213905 TI - Tweaking the tools of "quality" measurement. PMID- 16213906 TI - Systematic node dissection by VATS is not inferior to that through an open thoracotomy: a comparative clinicopathologic retrospective study. AB - BACKGROUND: Major pulmonary resection with systematic node dissection (SND) for early lung cancer by video-assisted thoracic surgery (VATS) is performed in many institutes, but the feasibility of SND for early lung cancer by VATS remains controversial. The aim of this study was to elucidate the feasibility and safety of SND by VATS. METHODS: Three hundred fifty patients with clinical stage I lung cancer who underwent pulmonary major resection with SND between 1998 and 2003 were enrolled in this study. Of these patients, 191 (VATS group) underwent pulmonary resection with SND by VATS; 159 patients (open thoracotomy [OT] group) did so through anterolateral thoracotomy. The clinical and pathologic data, including the number of dissected nodes in each nodal station, of the 2 groups were compared to evaluate the feasibility of SND by VATS. RESULTS: Pathologic data showed that, in the VATS group, more patients had adenocarcinoma (P = .0078) and fewer patients had advanced factors than the OT group. The greatest tumor diameter was 24.5 mm and 29.6 mm in the VATS group and OT group, respectively (P < .0001). The total number of mediastinal nodes dissected in right upper lobectomy plus right middle lobectomy (RUL+RML), right lower lobectomy (RLL), left upper lobectomy (LUL), and lower left lobectomy (LLL) also did not differ between the 2 groups. The total number of mediastinal nodes dissected in RUL+RML, RLL, LUL, and LLL was 19.7 in the VATS group versus 22.0 in the OT group (P = .122), 23.4 versus 21.0 (P = .241), 14.8 versus 17.5 (P = .123), and 18.8 versus 15.8 (P = .202), respectively. The number of dissected nodes in each nodal station in RUL+RML, RLL, LUL, and LLL was similar between the 2 groups. Operative mortality, morbidity, or recurrence did not differ between the 2 groups. CONCLUSIONS: With regard to the number of dissected nodes, SND by VATS was not inferior to that of OT. SND by VATS is technically feasible and safe, and seems acceptable for clinical stage I lung cancer. PMID- 16213907 TI - Hemodynamic changes of splenogastric circulation after spleen-preserving pancreatectomy with excision of splenic artery and vein. AB - BACKGROUND: The safety of spleen conservation without preservation of the splenic artery and vein was proved on the basis of short-term observation, but the long term results of this procedure have been uncertain. To clarify the hemodynamic changes of splenogastric circulation of patients undergoing spleen-preserving pancreatectomy with excision of the splenic artery and vein, we retrospectively analyzed patient outcome with particular reference to the assessment of hemodynamic changes of splenogastric circulation. METHODS: Ten patients who had undergone spleen-preserving pancreatectomy with excision of the splenic artery and vein were retrospectively analyzed. In all patients both the short gastric and left gastroepiploic arteries and veins were preserved. All patients were observed for a minimum of 52 months. Collateral venous pathways were evaluated by computed tomography and endoscopy. RESULTS: Early complications such as splenic infarction and atrophy did not occur in any of the patients, but computed tomography revealed perigastric varices in 7 patients (70%) and submucosal varices in 2 patients (20%). Endoscopy showed gastric varices in 2 patients in whom submucosal gastric varices were identified on computed tomography. Gastrointestinal bleeding from gastric varices occurred in 1 patient. In 1 patient without gastric varices, a gastrorenal shunt was demonstrated on computed tomography. CONCLUSIONS: This study confirmed that gastric varices frequently occurred in patients who underwent spleen-preserving pancreatectomy with excision of the splenic artery and vein. PMID- 16213908 TI - Rat gastric injury after lipopolysaccharide: role of inducible nitric oxide synthase. AB - BACKGROUND: Short-term treatment with lipopolysaccharide (LPS) causes morphologic, but not macroscopic, gastric injury and decreases gastric injury caused by a subsequent challenge with a luminal irritant. This effect is abrogated by inducible nitric oxide synthase (iNOS) inhibition. The effects of long-term treatment with LPS on gastric injury are unknown as is the role of iNOS. We hypothesized that LPS would cause macroscopic gastric injury at later time points through an iNOS-dependent pathway. METHODS: Conscious rats were given saline or LPS (1 or 20 mg/kg intraperitoneal) as a single intraperitoneal injection and killed 24 to 72 hours after injection. Macroscopic gastric injury (computerized planimetry), gastric luminal fluid volume and pH, and iNOS protein levels were assessed. RESULTS: When compared with saline, high-dose but not low dose LPS caused macroscopic gastric injury, increased gastric luminal fluid and pH, and up-regulated iNOS at 24 and 48 hours. All assessments returned to baseline by 72 hours. Inhibition of iNOS with 1400W (1 mg/kg intraperitoneal) given 15 minutes before saline or LPS (20 mg/kg) attenuated the deleterious effects of LPS on gastric injury and pH, but not fluid accumulation. CONCLUSIONS: These data suggest that prolonged treatment with high-dose LPS causes gastric injury through an iNOS-mediated pathway. PMID- 16213910 TI - A day in the life ... PMID- 16213909 TI - Ethyl pyruvate ameliorates ileus induced by bowel manipulation in mice. AB - BACKGROUND: Ethyl pyruvate (EP) improves survival, decreases proinflammatory cytokine expression, and ameliorates organ dysfunction in mice who have lethal sepsis or were subjected to hemorrhagic shock. Herein, we tested the hypothesis that treatment with EP can prevent the development of ileus after bowel manipulation, a phenomenon that is mediated by an inflammatory response in the bowel wall. METHODS: C57Bl/6 mice underwent operative manipulation of the small intestine or were subjected to a sham procedure. Some of the mice subjected to gut manipulation were pre- and post-treated with 4 doses of EP (40 or 80 mg/kg per dose), whereas others received similar volumes of the vehicle for EP. Gastrointestinal transit of a nonabsorbable marker was assessed by gavaging the mice with the tracer 24 hours after operation and assessing its concentration 90 minutes later in bowel contents from the stomach, 10 equally long segments of small intestine, the cecum, and 2 equally long segments of colon. The contractile responses of ileal circular smooth muscle to graded concentrations of bethanechol were assessed by using standard organ bath methodology. Expression of interleukin 6 and inducible nitric oxide synthase transcripts in ileal muscularis propria was assessed by using the semiquantitative reverse transcriptase-polymerase chain reaction. RESULTS: In sham-operated controls, the mean (+/- SE) geometric center for the transit marker was 10.0 +/- 0.5, whereas for vehicle-treated mice subject to bowel manipulation, the value for this parameter was 3.5 +/- 0.1 (P < .05). When mice subjected to bowel manipulation were treated with several 40 mg/kg doses of EP, the geometric center was 7.3 +/- 1.0 (P < .05 vs sham-operated group). Gut manipulation impaired intestinal smooth muscle contractility in vitro and increased steady-state levels of interleukin-6 and inducible nitric oxide synthase messenger RNA. Treatment with EP ameliorated these effects of gut manipulation. CONCLUSIONS: EP warrants further evaluation as a therapeutic agent to ameliorate postoperative ileus. PMID- 16213911 TI - Gallstone ileus of the colon. PMID- 16213912 TI - Re: "laparoscopic assisted distal gastrectomy for early gastric cancer: five years' experience". PMID- 16213913 TI - A problematic complication of sex change operation. PMID- 16213914 TI - Paracrine mediators of mouse uterine epithelial cell transepithelial resistance in culture. AB - Epithelial cell integrity at mucosal surfaces provides an effective physical barrier against potential pathogens that threaten reproductive health. We have used polarized epithelial cells from adult mouse uteri to investigate the roles of TNFalpha and TGFbeta, which are produced by uterine epithelial and stromal cells, and hepatocyte growth factor (HGF), produced by uterine stromal cells, in regulating epithelial cell integrity measured as transepithelial electrical resistance (TER). Exposure of epithelial cells to TNFalpha, TGFbeta, and HGF have profound effects on TER that are different from their known actions on TER at other mucosal surfaces. When incubated with TNFalpha, TER increased in a dose dependent manner. In contrast, when cells were incubated with TGFbeta, TER was markedly but reversibly suppressed. Interestingly, HGF, when placed in the basolateral compartment, increased TER. Based on these findings, we conclude that TNFalpha, TGFbeta and HGF may play regulatory roles in modulating epithelial cell tight junctions. These studies suggest that factors, such as hormone balance, pathogen exposure as well as pregnancy, which affect cytokine and growth factor secretion, influence epithelial cell barrier protection within the female reproductive tract. PMID- 16213915 TI - Optimal duration of allergic rhinitis clinical trials. AB - OBJECTIVE: Guidelines have been published by the Food and Drug Administration (FDA) and the European Agency for the Evaluation of Medicinal Products (EMEA) for the conduct of seasonal allergic rhinitis (SAR) and perennial allergic rhinitis (PAR) studies. These guidelines have differences regarding the duration of such trials: the FDA suggests 2 weeks for SAR and 4 weeks for PAR but the EMEA suggests 2 to 4 weeks for SAR and 6 to 12 weeks for PAR trials. In the interest of global harmonization, it would be desirable to have a uniform duration of such trials so that investigators, internationally, would be able to readily compare results for various types of treatments based on a single standard. Therefore, we performed an evidence-based review to answer the clinical question, What is the optimal duration for SAR and PAR clinical trials? METHODS: We performed a MEDLINE search of the published literature from 1995 to the present. We used appropriate search terms, such as allergic rhinitis, seasonal allergic rhinitis, perennial allergic rhinitis, SAR, and PAR, to identify pertinent articles. These articles were reviewed and graded according to the evidence quality. RESULTS: After an initial screening of more than 300 articles, 138 articles were analyzed thoroughly. No study specifically addressed the question of the optimal duration of SAR or PAR clinical trials. CONCLUSIONS: We conclude that the current FDA (draft) guidelines calling for a study length of 2 weeks for the assessment of drug efficacy for SAR and 4 weeks for the study of drug efficacy in PAR are appropriate and that longer study periods are not likely to add meaningfully to the assessment of drug efficacy. PMID- 16213917 TI - Sinus headache, migraine, and the otolaryngologist. AB - "Sinus headache" is a common complaint in the otolaryngology office. Recent literature has shown that the majority of patients with this complaint satisfy the diagnostic criteria for migraine. This review article presents an overview of the sinus headache literature, with emphasis on the incidence, identification, and treatment of migraine headache for the otolaryngologist. PMID- 16213918 TI - Analysis of battlefield head and neck injuries in Iraq and Afghanistan. AB - OBJECTIVE: At the time of this study, the 1st place that an injured or ill American soldier in Iraq or Afghanistan would have been evaluated by an ENT-head and neck surgeon was at a tertiary care medical center as a result of air evacuation out of theater: Landstuhl Regional Medical Center (LRMC), Ramstein, Germany. By examining the ENT-related diagnoses of all air evacuations from downrange, we were able to match the patients classified as having battle injuries to determine the percentage with head and neck trauma. STUDY DESIGN: A prospective review of 11,287 soldiers air-evacuated from Afghanistan and Iraq, representing the 1st year of combat operations. A new, computerized patient tracking system was created by our team to merge several disparate databases to generate and compile our data. RESULTS: The ENT-head and neck surgery department evaluated and primarily managed 8.7% of all patients evacuated out of theater by air to Germany. Other medical and surgical services managed 7.3% of all patients evacuated out of theater with overlapping ENT diagnoses. The number of potential ENT patients increased to 16% when one looked at all head and neck pathology instances seen by all medical and surgical departments hospitalwide. Of all patients air-evacuated and classified as having battle injuries, 21% presented with at least 1 head and neck trauma code. CONCLUSIONS: This is the 1st paper focusing on the role of the ENT-head and neck surgeon in treating a combat population and also the patterns of illness and head and neck injuries in a deployed force in our modern military. Improved soldier body armor has resulted in distinctly new patterns of combat injuries. Unprotected areas of the body account for the majority of injuries. SIGNIFICANCE: These findings should be used to improve the planning and delivery of combat medical care. PMID- 16213919 TI - Awareness of general practitioners towards treatment of laryngopharyngeal reflux: a British survey. AB - OBJECTIVE: Patients with acid reflux can occasionally present with atypical symptoms such as globus pharyngeus, constant throat clearing, chronic cough, hoarseness, catarrh, choking episodes or asthma-like symptoms. The aim of this survey was to determine whether general practitioners are aware of the atypical manifestations of reflux and the differences in treatment between laryngopharyngeal reflux and gastroesophageal reflux. DESIGN: Questionnaire Survey. SETTING: Primary Care RESULTS: One hundred and sixty general practitioners who routinely refer patients to our Department of Otolaryngology were selected and a postal survey was conducted. One hundred and fifty of these responded (94% response rate). The commonest symptoms for which proton pump inhibitors are prescribed are heartburn (65%), followed by a combination of heartburn and other symptoms (15%), chronic cough (4%), choking episodes (4%), asthma-like symptoms (3%), hoarseness (2%), globus (2%), catarrh (1%), dysphagia (1.5%), frequent throat clearing (1.5%), halitosis and/or bitter taste (1%). CONCLUSIONS: Our results suggest that the majority of the general practitioners surveyed are unaware of the entity laryngopharyngeal reflux or reflux symptom index. More awareness is required in the primary care setting for early recognition of patients with suspected laryngopharyngeal reflux. PMID- 16213920 TI - Expanding applications for the microdebrider in pediatric endoscopic airway surgery. AB - OBJECTIVE: The microdebrider has been described for treating laryngeal papillomas, but there are no reports of other uses for this instrument in the pediatric airway. The objective of this article is to describe expanding applications for the microdebrider in pediatric airway surgery. STUDY DESIGN AND SETTING: Retrospective review, tertiary care academic institution. RESULTS: The microdebrider was used in 75 pediatric airway procedures. Twenty-two of these cases involved novel, previously unreported applications of the microdebrider. CONCLUSIONS: The microdebrider can be used safely and effectively for a variety of pediatric airway pathologies, including papillomas, granulations, and stenoses from the larynx to the distal trachea. SIGNIFICANCE: Correct usage of the various tip configurations on the microdebrider has made management of many airway lesions commonly encountered by pediatric otolaryngologists safer, more expedient, and simpler than management by other methods. In our practice, this instrument largely has supplanted the use of the CO(2) laser and some open approaches for pediatric airway lesions. PMID- 16213921 TI - Surgical anatomy of the recurrent laryngeal nerve revisited. AB - OBJECTIVES: Previous anatomic studies of the recurrent laryngeal nerves (RLNs) have described the variability in the course of the RLN. The anatomy of the nerve appears more constant along its distal segment near the cricothyroid joint, which is our surgical approach to the initial identification of the nerve. Understanding the topographical anatomy of the nerve in this region facilitates quick and safe nerve identification. The surgical topographical anatomy of the nerve in this region has not been studied in detail, which is the focus of this study. METHODS: A total of 278 RLNs in 190 patients were dissected during thyroidectomy and/or parathyroidectomy. The course of the nerve was recorded, paying particular attention to the directional course along its distal portion. The angle in which it coursed in relationship to a line paralleling the tracheoesophageal groove was determined. RESULTS: All 278 nerves were identified. Seventy-eight percent of the right-sided nerves coursed between 15 and 45 degrees, and 77% of the left-sided nerves coursed between 0 and 30 degrees. It appears that the nerve is more likely to travel at a more obtuse angle with right sided RLNs and in patients with a low-lying cricoid. There was no permanent postoperative RLN palsy, and the incidence of temporary palsy was 1%. CONCLUSIONS: Approaching the nerve along its distal portion is safe and effective. The surgical topographical anatomy in this region is described in detail. Some of the potential advantages of identifying the nerve more distally include less chance of disrupting the blood supply to the inferior parathyroid gland, dissection along a shorter portion of the nerve, and less variability of the nerve. PMID- 16213922 TI - Powered instrumentation in the treatment of tracheal granulation tissue for decannulation. AB - OBJECTIVE: To evaluate the outcomes of powered instrumentation tracheal surgery (PITS) in patients with tracheal granulation tissue following tracheostomy that caused or exacerbated airway obstruction and impeded decannulation. STUDY DESIGN: Retrospective case review. METHODS: Sixteen patients that accepted PITS for decannulation from tracheal granulation tissue were reviewed. The surgical procedure was performed with the assistance telescope connected to a television monitoring system. The success of PITS was defined as a decannulation for 3 months without a recurrence of airway obstruction. RESULTS: The average operation time was 34 minutes. No complication was met in the perioperative period. Fourteen cases (87.5%) were successfully decannulated within 3 days following PITS. The 2 cases that failed successful decannulation had undergone insertion of a tracheal stent for their hindered tracheal stenosis. CONCLUSIONS: The preliminary results reveal that the new operation modality, PITS, is a safe and simple method for the treatment of tracheal granulation tissue and could be a useful alternative for benign tracheal lesions. PMID- 16213923 TI - The effects of previously acquired skills on sinus surgery simulator performance. AB - OBJECTIVE: The Endoscopic Sinus Surgery Simulator is currently being studied as a tool for otolaryngology resident training. While examining performance patterns on the simulator, we sought to define any relationships that may exist between students' previously acquired skills and their performance on the simulator. METHODS: Twenty-six medical students were enrolled in our study. After completing their trials, they were asked to complete a 28-item questionnaire. This questionnaire included yes/no questions as well as 10-point Likert scale instruments. RESULTS: Only 4 students did not respond to the questionnaire. Significant contributions to simulator performance were elucidated for a number of previously learned skills, but most were short-lived. For example, experienced video gamers were significantly better at hazard avoidance in the simulator's novice mode (P = .03), but their advantage dissipated as they advanced to intermediate mode trials. In addition, students' handedness contributed to simulator performance for only the first 3 trials (P = .04), but this relationship, too, was no longer statistically significant in later trials. CONCLUSIONS: Some study students had skills and characteristics that significantly aided them in their ability to perform surgery on the simulator. However, these benefits were short-lived, and these results suggest that practicing plays a more important and long-standing role than other factors in surgical simulator performance. It follows that surgical simulation should play a more prominent role in surgical education because residents who practice on simulators such as this are, in turn, practicing for the reality of the operating room. PMID- 16213924 TI - Radiation dose to the eyes and parotids during CT of the sinuses. AB - OBJECTIVE: To measure the radiation dose to the lens and parotid during high resolution computed tomography scan of the sinuses. STUDY DESIGN AND SETTING: Nine cadaver heads were scanned in the axial plane by means of a fine-cut (0.75 mm) protocol. Images were then reconstructed in the coronal and sagittal planes for use with the image guidance software. Thermoluminescent dosimeters were taped over the eyes and parotids and used to measure the radiation dose absorbed by these organs. RESULTS: Doses obtained were 29.5 mGy for the lens and around 30 mGy for the parotid. CONCLUSION: The measured doses are lower than the reported acute thresholds of 500-2000 mGy for lens opacities and well below the threshold of 2500 mGy for damage to the parotid. SIGNIFICANCE: These results demonstrate minimal risk from radiation through the use of high-resolution computed tomography and support the use of such a protocol for diagnosis and preoperative planning. PMID- 16213925 TI - Beef, pork, and milk allergy (cross reactivity with each other and pet allergies). AB - OBJECTIVE: The purposes of this study were to examine milk allergic patients to determine concomitant reactivity between milk, beef, pork and cat and dog dander and other common inhalant allergens. METHODS: 19 patients were selected according to their Immuno-CAP results, which had increased Ig-E levels against milk, pork or beef. Patients were also tested against Johnson grass, short ragweed, cat/dog dander and d. farina. RESULTS: Pearson's test revealed strong correlation between beef and pork, beef and milk, pork and milk Ig-E counts (consecutively r2 = 0.89, r2 = 0.81, r2 = 0.60 and p < 0.01. All cat allergic patients also appeared to be allergic to either beef/pork meat or milk. The correlation between pork and dog dander Ig-E counts was also significant (r2 = 0.38, p < 0.01). No correlation detected between milk-meat-pet and grass-weed-dust allergies. DISCUSSION AND CONCLUSION: Patients who are known to have pet allergies may need to be screened for meat and milk allergy. Milk allergic patients may also need to avoid cows and pork meat. PMID- 16213926 TI - The effects of short-term immunotherapy using molecular standardized grass and rye allergens compared with symptomatic drug treatment on rhinoconjunctivitis symptoms, skin sensitivity, and specific nasal reactivity. AB - BACKGROUND: The efficacy and safety of short-term immunotherapy with molecular standardized allergens (STI) has been demonstrated by double-blind placebo controlled clinical trials. The aim of this study was to compare STI with symptomatic drug treatment. METHODS: Forty-eight patients with rhinoconjunctivitis to grass and/or rye pollen were treated either with STI (ALK(7), n = 24) plus anti-allergic drugs or anti-allergic drugs, alone (n = 24) in a prospective, randomized study. Symptoms and use of drugs were reported in patient diaries and titrated nasal provocation and skin prick tests were performed at baseline, before, and after season. RESULTS: Median overall symptom (P = 0.022, U test) and medication scores (P = 0.003) were significantly lower in the STI group, as was the result for a simultaneous analysis of conjunctival, nasal, and bronchial symptom scores and medication (P = 0.005). Sensitivity in the nasal provocation test decreased in the STI group but not in the drug-treated group. These differences became significant directly after STI (P = 0.027) as well as after the grass pollen season (P < 0.001). Skin sensitivity did not change in the STI group but increased in the drug-treated group after season, with a significant difference between the two groups for the erythema (P < 0.001). CONCLUSIONS: STI reduces grass pollen-induced rhinoconjunctivitis symptoms and drug use, and specific nasal reactivity and skin sensitivity, more efficiently than a standard symptomatic treatment. PMID- 16213927 TI - Isolated sphenoid sinus disease: etiology and management. AB - OBJECTIVE: To evaluate the diagnosis and management of isolated sphenoid sinus disease by using the current rhinologic standard of care. STUDY DESIGN: Retrospective chart review. RESULTS: Fifty sequential, symptomatic patients were studied. Presenting symptoms included headache or facial pain (88%), rhinorrhea (46%), and nasal congestion (26%). All patients underwent CT imaging, demonstrating bony changes or dehiscences (42%), a mass (24%), or complete opacification of the sphenoid sinus (22%). Eighty percent required surgical intervention. The most frequent diagnoses were as follows: sinusitis (38%), fungal ball (20%), neoplasm (16%), and mucocele (12%). Treatment resulted in clinical or endoscopic improvement or resolution in 87% of the patients. CONCLUSION: The presenting symptoms of isolated sphenoid sinus disease can be nonspecific and may result in an inordinate delay in diagnosis. Nasal endoscopy and radiologic imaging are central to making an accurate and timely diagnosis. Medical treatment or minimally invasive surgical techniques can successfully manage the majority of patients with persistent or refractory symptoms. PMID- 16213928 TI - Endoscopic frontal sinus obliteration: a new technique for the treatment of chronic frontal sinusitis. AB - OBJECTIVES: Patients who fail endoscopic drainage procedures for chronic frontal sinusitis often require obliteration of the frontal sinus with abdominal fat. The purpose of this study was to evaluate an endoscopic technique for frontal sinus obliteration. STUDY DESIGN AND SETTING: Retrospective case-control. Thirty-five patients underwent frontal sinus obliteration using either an endoscopic (n=10) or conventional osteoplastic flap (n=25) technique from 1994 to 2004 at an academic medical center. RESULTS: Patients undergoing endoscopic obliteration had less blood loss (P = 0.006), decreased operative time (P = 0.016), and a shorter hospital stay (P = 0.003) compared to osteoplastic control subjects. All 3 surgical complications occurred in the control group. No patients required additional surgery for frontal sinusitis. CONCLUSIONS: The endoscopic approach to frontal sinus obliteration appears to reduce patient morbidity and should be considered in the surgical management of advanced frontal sinus disease. SIGNIFICANCE: This is the first report of a minimally-invasive technique for frontal sinus obliteration. PMID- 16213929 TI - Does increased electrocautery during adenoidectomy lead to neck pain? AB - OBJECTIVES: The objective was to assess the impact of electrocautery on complications in adenoidectomy. We sought to quantify cautery-related temperature changes in prevertebral fascia that may occur during the procedure, retrospectively evaluate the incidence of cautery-related complications, and prospectively assess the role of cautery in postoperative neck pain. METHODS: Three consecutive related trials were performed. Initially, adenoidectomy was performed on 20 fresh cadavers, using a thermister to evaluate temperature changes in the prevertebral fascia after electrocautery (30 watts over a 30 second period). Next, retrospective analysis of adenoidectomy complications in 1206 children over a 5-year period was performed. Based on these findings, a prospective study of the incidence of neck pain following adenoidectomy was performed in a cohort of 276 children. Adenoidectomy technique, wattage, and duration of electrocautery were recorded for each child. Children with significant neck pain were evaluated with MRI. RESULTS: Peak thermister readings averaged 74 degrees C, for a mean change of 51.8 degrees C. Complications observed in retrospective analysis included neck pain (3), Grisel's syndrome (1), prolonged velopharyngeal insufficiency (1), retropharyngeal edema (1), and severe nasopharyngeal stenosis (1). The incidence of neck pain in the prospective study was 12% (33 pts), and was independent of adenoidectomy technique, cautery wattage, or duration of cautery use. MRIs revealed edema without abscess. CONCLUSIONS: Cautery can result in substantial temperature changes in the surgical adenoid bed. Despite this, the incidence of complications, specifically neck pain, associated with adenoidectomy is low, although underreported. Complications appear to be independent of adenoidectomy technique and cautery use. PMID- 16213930 TI - Antimicrobial therapy for children with adenotonsillar hypertrophy and obstructive sleep apnea: a prospective randomized trial comparing azithromycin vs placebo. AB - OBJECTIVE: Multiple clinical trials demonstrate the value of administering antibiotics for recurrent tonsillitis. However, there is no consensus as to the role of antibiotics in the management of adenotonsillar hypertrophy and obstructive sleep apnea (OSA). It has been suggested that antibiotics may reduce adenotonsillar size, improve obstructive symptoms, and obviate the need for surgery. The goal of this pilot study was to evaluate the efficacy of a broad spectrum antibiotic in the management of adenotonsillar hypertrophy and OSA. METHODS: Twenty-two children ages 2 through 12, with evidence of OSA, were enrolled in this double-blinded, prospective clinical trial. Children were randomly assigned into 1 of 2 groups. Group I received azithromycin for a 30-day period (12 mg/kg on days 1-5; the regimen was repeated on days 11-15 and days 21 25). Group II received a placebo administered in the same regimen for a 30-day period. Overnight polysomnography was performed before and within 2 weeks after drug administration. RESULTS: Polysomnographic indices of OSA severity appear to indicate an improvement in the antibiotic as compared to the placebo group, but a comparison of mean +/- standard error for each parameter failed to demonstrate statistical significance using the paired t test. Only 1 of 11 (9%) children had normalization of their polysomnographic parameters. CONCLUSIONS: These results suggest that a course of a broad-spectrum antibiotic may be effective in temporarily improving OSA due to adenotonsillar hypertrophy, but does not appear to obviate the need for surgery. PMID- 16213931 TI - Quality of life after adenotonsillectomy for SDB in children. AB - OBJECTIVE: To evaluate the relationship between quality of life and the relative severity of sleep-disordered breathing (SDB) and to compare changes in quality of life after adenotonsillectomy in children with similar demographics but with either obstructive sleep apnea syndrome (OSAS) or with milder forms of SDB. STUDY DESIGN AND SETTING: All study participants underwent polysomnography to document the severity of SDB. The effectiveness of adenotonsillectomy for the relief of SDB was evaluated by using the OSA-18 quality of life survey (OSA-18). Preoperative and postoperative OSA-18 scores for each group of children (OSAS and mild SDB) were compared by using a repeated measures ANOVA. Changes between the 2 groups were compared by using an analysis of covariance with the preoperative score as a covariate. RESULTS: The study population included 61 children, 43 with OSAS and 18 with mild SDB. The demographics in the 2 groups were similar. The mean apnea-hypopnea index for children with OSAS was 21 (range, 5-46), and for children with mild SDB, it was 3 (range 0-4.9). The total OSA-18 score and the scores for all domains showed significant improvement after surgery for both groups of children (P < .001). A comparison of mean difference in total and domain scores for the 2 groups of children was not significant. CONCLUSIONS AND SIGNIFICANCE: Preoperative values for the OSA-18 total and domain scores are high in children with either OSAS or mild SDB. Both groups of children show a dramatic improvement in quality of life after adenotonsillectomy and the degree of improvement is similar. Fortunately, surgical therapy with adenotonsillectomy is associated with marked improvement in quality of life for children with either OSAS or mild SDB. PMID- 16213932 TI - Long-term results of palatal implants for primary snoring. AB - OBJECTIVE: To determine the safety and efficacy of the Pillar Palatal Implant System over a 1-year follow-up period. STUDY DESIGN AND SETTING: In this prospective study, 40 healthy adult patients with primary snoring due to palatal flutter were treated after clinical, polysomnographic, and endoscopic examination. Under local anesthesia 3 implants were placed into the soft palate. Postoperative morbidity and functional parameters were assessed. Snoring and daytime sleepiness were assessed before and 90, 180, and 360 days after surgery. Objective data were obtained by polysomnography and SNAP-recording before and 90 days after treatment. RESULTS: All implants were placed without complications. A total of 13 implants partially extruded uneventfully in 10 patients. Functional parameters remained unchanged. After 1 year, snoring was reduced from 7.1 +/- 1.9 to 4.8 +/- 2.5 (Visual-Analogue-Scale, P < 0.05) and daytime sleepiness from 6.1 +/- 3.2 to 4.9 +/- 3.1 (Epworth-Sleepiness-Scale, P < 0.05). SNAP data and polysomnography parameters showed clinically irrelevant changes. CONCLUSION: Our data demonstrate a significant decrease in snoring and daytime sleepiness over a period of one year. PMID- 16213933 TI - A prospective randomized single-blind trial comparing ultrasonic scalpel tonsillectomy with tonsillectomy by blunt dissection in a pediatric age group. AB - OBJECTIVE: To determine if there was a difference in postoperative pain and dietary intake when comparing ultrasonic scalpel (US) with blunt dissection tonsillectomy (BD) in children. STUDY DESIGN AND SETTING: A prospective single blind randomized controlled trial comparing US and BD tonsillectomy in children aged between 5 and 13 years who undergo tonsillectomy for recurrent acute tonsillitis in a specialized pediatric hospital. RESULTS: One hundred and twenty two children were enrolled. Ninety-three (76%) patients completed dietary and pain scales. Pain was significantly greater in the US group on postoperative days 1 and 3 (P = 0.0466, P = 0.0084) and overall (P = 0.0003). Dietary intake scores were significantly better in the US group on days 1, 5, 7, and 9, however, these scores were inconsistent with dietary intake scores declining in both groups throughout the study. Blood loss was statistically significantly greater in the BD group (33.1 ml vs 3.0 ml, P < 0.0001). There were no significant differences for all other observed outcomes. CONCLUSIONS: US tonsillectomy causes significantly more pain overall and on postoperative days 1 and 3 when compared to BD tonsillectomy. US tonsillectomy causes less intraoperative blood loss, the clinical significance of which is questionable. SIGNIFICANCE: US tonsillectomy appears to cause more postoperative pain than BD tonsillectomy in children aged 5 to 13 years undergoing tonsillectomy for recurrent acute tonsillitis. PMID- 16213934 TI - Microbiology of acute otitis media with tympanostomy tubes. AB - OBJECTIVE: The objective was to determine the types of organisms which cause acute otitis media with a tympanostomy tube and to ascertain their frequency distribution. STUDY DESIGN AND SETTING: Prospective, randomized, multi institutional clinical trials. Both private and academic sites were included. RESULTS: 1309 isolates were recovered from 956 draining ears. Streptococcus pneumonia was recovered from 17%, Staphylococcus aureus from 13%, H flu from 18% and Pseudomonas aeruginosa from 12%. Fungal organisms were recovered from 5% of total isolates and 4% from single isolates. CONCLUSIONS: AOMT is microbiologically different than AOM with an intact TM. There is no evidence that resistance develops as result of topical treatment. SIGNIFICANCE: The study demonstrates that AOMT is frequently caused by organisms not susceptible to oral antibiotics approved for children, but which are sensitive to topical ear drops. PMID- 16213935 TI - Experimental and clinical experience of albumin coating of tympanostomy tubes. AB - OBJECTIVE: Otorrhea and tube occlusion are typical problems with tympanostomy tubes (TT). The purpose of this study was to test glue protein, fibronectin adhesion on albumin-coated and uncoated TT surfaces and to show the effect of this method on TT sequelae in vivo. STUDY DESIGN AND SETTINGS: Fibronectin binding on TT surface was tested in two in vitro experiments. Thereafter 170 patients were randomized in a prospective clinical trial to test the effect of the method in vivo. The extruded TTs that could be collected from ears of some study patients were imaged with scanning electron microscopy (SEM). RESULTS: Marked binding inhibition of fibronectin on albumin-coated TTs was found. Fewer tube sequelae were found in ears with albumin-coated TTs. SEM revealed thick crusts on uncoated tube surface. CONCLUSION: Albumin coating of TTs can reduce unwanted adhesion on the tube surface in vitro and tube occlusions in vivo. SIGNIFICANCE: TT sequelae can be reduced by surface coating with albumin. PMID- 16213936 TI - Laser-assisted tympanostomy in pediatric patients with serous otitis media. AB - OBJECTIVES: To evaluate prognostic factors related with cure rate, in pediatric patients with serous otitis media treated with laser-assisted tympanostomy without ventilation tubes, in a single institution. PATIENTS AND METHOD: The procedure was performed on 124 ears in 88 individuals, from 3 to 14 years old. External auditory canal anatomy, type of anesthesia, tympanic membrane and middle ear fluid characteristics, myringotomy size, and laser parameters, in relation to cure rate, were accordingly studied. RESULTS: The overall cure rate by ear at the end of the 2-month follow-up period was 54.83%, whereas 45.17% still suffered from otitis media. Multivariate statistical analysis demonstrated that the presence of a thick tympanic membrane is significantly correlated with pure outcome in children with serous otitis media, when laser-assisted tympanostomy without ventilation tubes is performed favoring a worse cure rate (P < 0.023). Other parameters did not statistically correlate with the outcome. A 41% parental dissatisfaction rate was noticed. CONCLUSION: This study addresses selection bias for children with serous otitis media, candidates for laser-assisted tympanostomy. These are related to the duration of serous otitis media, the condition of middle ear mucosa, the thickness of the tympanic membrane, the type of anesthesia, and the cost of laser apparatus. There is likely to be a causal relationship between outcome and tympanic membrane appearance in children undergoing laser-assisted tympanostomy. PMID- 16213937 TI - Loss of reflex tearing: an expected consequence of juvenile nasopharyngeal angiofibroma. AB - OBJECTIVE: To report the loss of reflex tearing after surgical treatment of juvenile nasopharyngeal angiofibroma (JNA). STUDY DESIGN AND SETTING: A retrospective case series of 8 patients with surgical extirpation of JNA from 1995 to 2003 in a major teaching hospital setting was studied for symptomatic xerophthalmia. RESULTS: One patient was lost to follow-up. Four of the remaining 7 patients reported a dry ipsilateral eye after surgical treatment of JNA. CONCLUSION: The location of the pterygopalatine ganglion and its associated fibers in the pterygopalatine fossa is directly adjacent to the location of origin of JNA. Lacrimal innervation passes through the pterygopalatine ganglion. Given the extensive nature of advanced JNA and mandate for complete surgical excision, lacrimal dysfunction should be considered an expected consequence of surgery. SIGNIFICANCE: The loss of reflex tearing has not been reported as a consequence of JNA or its surgical treatment. EBM RATING: C. PMID- 16213938 TI - Combination docetaxel plus vitamin D(3) as an immune therapy in animals bearing squamous cell carcinomas. AB - OBJECTIVE: Background tumor growth results in the mobilization of immune inhibitory CD34(+) progenitor cells. However, vitamin D(3) can differentiate the CD34(+) cells into immune stimulatory dendritic cells. This study determined if docetaxel treatment could increase the impact of the vitamin D(3) to generate dendritic cells. METHODS: The murine squamous cell carcinoma model, SCC VII/SF, which is often used as a head and neck cancer model, was used to determine the immunological effects of two cycles of docetaxel plus vitamin D(3). RESULTS: Vitamin D(3) with or without docetaxel was similarly effective in reducing CD34(+) cell levels within the spleen, lymph nodes, and tumor. Dendritic cell levels were similarly enhanced in the lymph nodes by vitamin D(3) alone or combined with docetaxel. However, the combination treatment caused a prominent increase in intratumoral levels of active T cells, which was not observed by the individual treatments. CONCLUSION: Incorporating docetaxel treatment with vitamin D(3) differentiation-inducing treatment enhances intratumoral immune responsiveness. PMID- 16213939 TI - Electrocochleography is more sensitive than distortion-product otoacoustic emission test for detecting noise-induced temporary threshold shift. AB - OBJECTIVE: We investigated and compared the usefulness of the electrocochleography and distortion product otoacoustic emission tests for detecting the earliest noise-induced damage by analyzing the sensitivity and specificity of the 2 tests. STUDY DESIGN: A prospective study. METHODS: After listening to music at 90.3 +/- 4.2 dB in the same night-club for 2 hours continuously, 23 healthy normal ears experienced a temporary threshold shift exceeding 5 dB. Pure-tone audiometry, the distortion product otoacoustic emission test, and electrocochleography were performed before, immediately after, and 24 hours after the exposure. RESULTS: Before exposure, the measured distortion product/noise floor was 9.8 +/- 10.4, 23.5 +/- 6.4, 18.7 +/- 6.4, and 19.1 +/- 5.6 dB sound pressure level (SPL) at frequencies of 1, 2, 3, and 4 kHz, respectively. Immediately after exposure, it decreased significantly at 2, 3, and 4 kHz to 16.6 +/- 7.6, 12.5 +/- 6.8, and 14.8 +/- 7.7 dB SPL, respectively. Marked increases in the amplitude of the summating potential and summating potential/action potential ratio were recorded from 0.15 +/- 0.06 to 0.32 +/- 0.11 and 0.23 +/- 0.06 to 0.44 +/- 0.08, respectively. The respective sensitivity and specificity of electrocochleography were 76.7% to 88.5% and 91.0% to 100%. Those of the distortion product otoacoustic emission test were 54.8% to 62.2% and 75.5% to 87.0%, respectively. CONCLUSION: Electrocochleography appears to provide more sensitive and specific information than the distortion product otoacoustic emission test for detecting a noise-induced temporary threshold shift. PMID- 16213940 TI - Surgical treatment of retraction pocket with bone pate: scutum plasty for cholesteatoma. AB - OBJECTIVES: Postoperative attic retraction pocket is a critical issue and may be indicative of disease recurrence after canal-up tympanoplasty for middle ear cholesteatoma. Although scutum plasty is an acceptable procedure that can be used to prevent the pocket formation, how the scutum is reconstructed has remained controversial. The aim of this study was to clarify the value of scutum plasty using bone pate. STUDY DESIGN: Retrospective study. SETTING: Bone pate scutum plasty was performed in 69 ears during first-stage operations. We attempted to reconstruct a smooth attic bony wall. At the time of the second-stage operations, the results of the scutum plasty were examined. RESULTS: The incidence of retraction pocket was statistically reduced from 20.2% to 5.8%. CONCLUSION: Although the reconstructed scutum showed atrophy to some extent, bone pate scutum plasty is effective in preventing postoperative retraction pocket if a smoothly shaped attic wall can be reconstructed. PMID- 16213941 TI - Novel technique of silastic implant carving for thyroplasty type I surgery. PMID- 16213942 TI - Preventing graft failures anterior to the handle of the malleus: a modified surgical technique for underlay grafting. PMID- 16213943 TI - Tympanic membrane perforation and retained metal slag after a welding injury. PMID- 16213944 TI - Frontonasal dysplasia. PMID- 16213946 TI - Mesenchymal chondrosarcoma of the nasopharynx. PMID- 16213945 TI - An unusual foreign body in the larynx in an adult. PMID- 16213947 TI - Carcinoma cuniculatum: verrucous carcinoma of the skin of the face. PMID- 16213948 TI - A case of relapsing polychondritis with patulous eustachian tube syndrome. PMID- 16213949 TI - Tuberculosis of the parotid gland. PMID- 16213950 TI - About the existence of a foraminal laryngeal cyst. PMID- 16213951 TI - Liposarcoma of the cervico-nuchal region. PMID- 16213952 TI - A case of nasal sarcoidosis involving intraturbinate medulla complicated with allergic rhinitis. PMID- 16213953 TI - Spontaneous recovery of hearing in a cochlear implanted patient: a case that needs answers. PMID- 16213954 TI - Descending necrotizing mediastinitis following lingual tonsillectomy and elective tracheotomy. PMID- 16213955 TI - Juvenile myofibroma of the neck. PMID- 16213956 TI - Fibrous dysplasia of the temporal bone: assessment by three-dimensional helical CT imaging. PMID- 16213957 TI - Facial paralysis in a patient with cholesterol cyst of the petrous bone. PMID- 16213958 TI - Multiple myeloma involving the cricoid cartilage. PMID- 16213959 TI - Acute extratemporal facial nerve paralysis after a suicide attempt by hanging. PMID- 16213960 TI - Intranasal extrusion of the endovascular coil after occluding internal carotid artery for massive nasopharyngeal bleeding. PMID- 16213961 TI - Fourth branchial arch anomaly and pyriform sinus fistula as a rare cause of recurrent retropharyngeal abscess and thyroiditis in an adult. PMID- 16213962 TI - Penetrating stab injury of the sphenoid sinus and the brain. PMID- 16213963 TI - Acute pharyngitis: an unusual presentation of acute endemic typhus. PMID- 16213964 TI - Giant cornu cutaneum. PMID- 16213965 TI - Giant dermoid cyst of the petrous apex. PMID- 16213966 TI - Pilomatricoma: a late onset case in the neck. PMID- 16213967 TI - Metastatic parathyroid carcinoma in the superior vena cava. PMID- 16213968 TI - Laryngeal zoster mimicking a laryngeal cancer. PMID- 16213969 TI - Small cell carcinoma of the larynx: a case report. PMID- 16213970 TI - Extranasopharyngeal angiofibroma originating from the nasal septum. PMID- 16213971 TI - Vestibular dilatation in branchio-oto-renal syndrome. PMID- 16213972 TI - Cautery-assisted palatal stiffening operation for the treatment of obstructive sleep apnea syndrome. PMID- 16213975 TI - Seeking safety and security. PMID- 16213977 TI - Cynthia Kilburn Kelly--friend and mentor. PMID- 16213978 TI - The new fear factor: hazardous drugs revealed. PMID- 16213979 TI - Food and supplement safety. PMID- 16213981 TI - Older adults living in the community: issues in home safety. PMID- 16213982 TI - Health, work, and safety of farmers ages 50 and older. AB - Over half of America's 1.9 million farms are operated by persons over age 55. Farming is one of the most dangerous occupations in the nation; the oldest age group in farming suffers disproportionately high mortality and morbidity rates. Results from this study indicate that farmers may have unique perspectives of health and the role that work plays in their health outcomes. Despite the perceived positive attributes of farm work, the nature of the work performed by this sample of 725 farmers indicates the continued performance of tasks that place them at high risk for injury. Because of the contextual nature of farm life, work modification counseling may be more beneficial for the aging farmer than abstinence from work. Examples are provided. PMID- 16213983 TI - The clinical laboratory improvement amendment: how it applies to the use of urine dipsticks in the long-term care facility. AB - The urine dipstick is one of the waived tests that is used in the long-term care facility under guidelines of the Clinical Laboratory Improvement Amendment (CLIA). Through recent surveys, the Centers for Medicare and Medicaid Services (CMS), which oversee this program, have identified significant problems in many testing sites. Because of the magnitude of the problem, the surveys will be continued. To comply with CLIA guidelines and safely and appropriately use the urine dipstick in the long-term care facility, nurses need to be aware of these guidelines. The purpose of this article is to discuss CLIA requirements regarding waived tests, specifically the urine dipstick, in the long-term care facility. PMID- 16213984 TI - A case study: inappropriate use of amitriptyline in the elderly. AB - In 1999, nearly 7 million elderly persons were prescribed potentially inappropriate medications as identified by Beer's Criteria. Amitriptyline is among the most frequently inappropriately prescribed drugs according to Beer's Criteria. Interventions to educate prescribers on Beer's Criteria are indicated to improve appropriate prescribing to the elderly population. Nurses also need to be aware of the potential adverse effects of these drugs when providing care to geriatric patients. PMID- 16213985 TI - A new option in the treatment of skin tears for the institutionalized resident: formulated 2-octylcyanoacrylate topical bandage. AB - Skin tears are a common phenomenon in elderly institutionalized adults (EIAs). Incidence ranges from 0.92 to 2.5 per person/year. Little supportive literature exists regarding optimal treatment with many regimens reported. A convenience sample of 20 patients with Payne-Martin Category II and III skin tears of less than 8 hours' duration were prospectively evaluated with the use of a formulated 2-octylcyanoacrylate topical bandage. Patients were followed weekly until the tear healed. Complete healing occurred with 1 application of 2-OTB in 90% (18/20) of study subjects; 5% (n = 1) reported transient mild pain (less than 15 seconds), and 90% (n = 19) reported no pain. There were no incidents of cellulitis or infection. Shower and bathing routines were not interrupted. Cost averaged less than $1 per application. Clinician time averaged 1.5 minutes per application. Clinicians reported high satisfaction because repeated dressing changes were eliminated. PMID- 16213986 TI - Bipolar disorder: an uncommon illness? Recognizing and caring for the elderly person with bipolar disorder. AB - Bipolar disorder (BPD), an affective mood disorder formerly called manic depressive illness, is a diagnosis rarely seen in elders. It has components of major depression and sometimes mania or hypomania. Many elders previously diagnosed with schizophrenia in their past are now found to have the elements of BPD. The psychiatric community has become aware that bipolar disorder in elders is much more common than previously thought, and progress is being made in appropriate diagnosis and treatment of this condition. PMID- 16213988 TI - Melatonin inhibits neural apoptosis induced by homocysteine in hippocampus of rats via inhibition of cytochrome c translocation and caspase-3 activation and by regulating pro- and anti-apoptotic protein levels. AB - In the present study, we examined the molecular mechanism by which homocysteine causes neuronal cell apoptosis. We further investigated the mechanisms of melatonin's ability to reduce homocysteine-induced apoptosis. Consistent with its antioxidant properties, melatonin reduced homocysteine-induced lipid peroxidation and stimulated glutathione peroxidase enzyme activity in hippocampus of rats with hyperhomocysteinemia. Furthermore, melatonin treatment diminished cytochrome c release from mitochondria and reduced caspase 3 and caspase 9 activation induced by hyperhomocysteinemia. Chronic hyperhomocysteinemia also led to poly(ADP ribose) polymerase cleavage and subsequently DNA fragmentation. Treatment with melatonin markedly inhibited poly(ADP-ribose) polymerase cleavage and reduced DNA damage. Hyperhomocysteinemia caused an elevation of pro-apoptotic Bax levels while reducing anti-apoptotic protein, Bcl-2, levels. Daily administration of melatonin up-regulated Bcl-2 and down-regulated Bax levels. We propose that, in addition to its antioxidant properties, melatonin has the ability to protect neuronal cells against apoptosis mediated homocysteine neurotoxicity by modulating apoptosis-regulatory proteins in the hippocampus of rats. PMID- 16213987 TI - Region-specific growth properties and trophic requirements of brain- and spinal cord-derived rat embryonic neural precursor cells. AB - To determine whether neural precursor cells have region-specific growth properties, we compared the proliferation, mitogenicity, and differentiation of these cells isolated from the embryonic day 16 rat forebrain and spinal cord. Neural precursor cells isolated from both regions were cultured in growth medium supplemented with epidermal growth factor, basic fibroblast growth factor, or epidermal growth factor+basic fibroblast growth factor. Under all three conditions, both neural precursor cell populations proliferated for multiple passages. While spinal cord-derived neural precursor cells proliferated moderately faster in epidermal growth factor-enriched growth medium, brain derived cells proliferated much faster in basic fibroblast growth factor-enriched growth medium. When exposed to both epidermal growth factor and basic fibroblast growth factor, the two neural precursor cell populations expanded and proliferated more rapidly than when exposed to a single factor, with brain derived neural precursor cells expanding significantly faster than spinal cord derived ones (P<0.0001). Differentiation studies showed that both neural precursor cell populations were multi-potent giving rise to neurons, astrocytes, and oligodendrocytes. However, neuronal differentiation from brain-derived neural precursor cells was greater than spinal cord-derived ones (11.95+/-5.00% vs 1.92+/-1.13%; passage 2). Further, the two neural precursor cell populations differentiated into a similar percentage of oligodendrocytes (brain: 8.66+/ 5.85%; spinal cord: 7.69+/-3.91%; passage 2). Immunofluorescence and Western blot studies showed that neural precursor cells derived from both regions expressed receptors for basic fibroblast growth factor and epidermal growth factor. However, brain-derived neural precursor cells expressed higher levels of the two receptors than spinal cord-derived ones in growth medium containing epidermal growth factor+basic fibroblast growth factor. Thus, our results showed that neural precursor cells isolated from the two regions of the CNS have distinct properties and growth requirements. Identifying phenotypic differences between these neural precursor cell populations and their growth requirements should provide new insights into the development of cell therapies for region-specific neurological degenerative diseases. PMID- 16213989 TI - Sympathetic and T helper (Th)2 bias may ameliorate uterine fibroids, independent of sex steroids. AB - We propose that inadequate sympathetic bias and Th2 bias in the uterine environment contributes to the formation of fibroids, independent of the sex steroid status. We also propose that fibroids represent a modern maladaptation that partly results from decreasing exposure to seminal fluid, which contains catecholalmines, transforming growth factor beta1 (TGFbeta1), aldosterone, prostaglandins, and other factors that shift the uterine environment to sympathetic and T helper (Th)2 bias. Lower risk of fibroids is associated with pre-menarche, post-menopause, pregnancy, exposure to contraceptives, smoking, earlier age of first pregnancy, shorter interval since last pregnancy, higher parity, and non-obesity. These associations are currently attributed to alterations of sex steroids. However, the association may also be explained by the observation that pre-menarche, post-menopause, pregnancy, and smoking represent periods of sympathetic and Th2 bias. Furthermore, use of contraceptives, early age of first pregnancy, short interval since last pregnancy, high parity, abnormal pap smear, and non-obesity may represent surrogates for increased sexual activity and increased exposure to seminal fluid. Catecholalmines, aldosterone, TGF, and prostaglandins are among the seminal fluid components that promote sympathetic and Th2 bias. Vasectomized copulations protect against fibroids, an observation that undermines the steroid hypothesis and supports our hypothesis. The putative mechanism of action of uterine artery embolization (UAE) for fibroid treatment is starvation of blood supply, but the extensive collaterals that protect uterine perfusion would presumably also buffer against fibroid hypoperfusion. Instead, the sympathetic and Th2 responses to UAE related ischemia may contribute to fibroid regression. A potential explanation for the association of fibroids with intrauterine devices may be a Th1 cell mediated immune response to the foreign body, which may also enhance the contraceptive effect. Novel methods of preventing and treating fibroids by promoting sympathetic and Th2 shift through natural, pharmacologic, and neuromodulatory means are envisioned. Fibroids are likely a modern dysfunction given the high Darwinian fitness cost of fibroid-related infertility, and may be attributable to reduced intercourse frequency. Fibroids have been observed among animals in captivity that are presumably reproductively isolated. PMID- 16213990 TI - Follicular fluid concentrations of free insulin-like growth factor (IGF)-I during follicular development in mares. AB - The objective of the present study was to evaluate changes in concentrations of free insulin-like growth factor (IGF)-I in follicular fluid (FFL) during follicle development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6-15 mm; n = 54), medium (16-25 mm; n = 23) or large (>25 mm; n = 15) and FFL was collected. Free IGF-I levels in FFL in large follicles of follicular phase mares were greater (P < 0.05) than in large follicles of luteal phase mares and small or medium follicles of luteal and follicular phase mares. Free IGF-I concentrations were greater (P < 0.05) in large follicles of luteal phase mares than small but not medium follicles of luteal phase mares. FFL ratio of estradiol:progesterone paralleled changes in free IGF-I. Free IGF-I concentrations were negatively correlated (P < 0.05) with insulin-like growth factor binding protein (IGFBP)-2, -4 and -5 but not IGFBP-3 levels. In addition, free IGF-I concentrations in FFL were positively correlated (P < 0.01) with FFL estradiol, progesterone, androstenedione, estradiol:progesterone ratio, total IGF I and total IGF-II. We conclude that increases in intrafollicular levels of bioavailable (free) IGF-I are associated with increased steroidogenesis in developing mare follicles. PMID- 16213991 TI - Hormonal regulation of leptin receptor expression in primary cultures of porcine hepatocytes. AB - A study was conducted to elucidate hormonal control of leptin receptor gene expression in primary cultures of porcine hepatocytes. Hepatocytes were isolated from pigs (52 kg) and seeded into collagen-coated T-25 flasks. Monolayer cultures were established in medium containing fetal bovine serum for 1 day and switched to a serum-free medium for the remainder of the 3-day culture period. To establish basal conditions hepatocytes were maintained in serum-free William's E medium containing 10 nM dexamethasone and 1 ng/ml insulin. For the final 24 h, insulin (1 or 100 ng/ml) or glucagon (100 ng/ml), were added in the presence or absence of 100 nM triiodothyronine (T3). RNA was extracted and quantitative RT PCR was performed with primers specific for the long form and total porcine leptin receptors. Leptin receptor expression was calculated relative to co amplified 18S rRNA. Expression of the long form of the leptin receptor was confirmed under basal conditions. Insulin, glucagon and synthetic human proteins (ghrelin and GLP-1) at 100 ng/ml had no influence on leptin receptor expression; the addition of T3 was associated with a marked increase (P < 0.001) in expression of total and long forms of the leptin receptor by 1.6 and 2.4-fold, respectively. Addition of leptin to cells which were pre-treated with T3 for 24 h (to up-regulate leptin receptor expression), confirmed the lack of a direct effect of leptin on glucagon-induced glycogen turnover and cAMP production. These data suggest that porcine hepatocytes may be insensitive to leptin stimulation even when leptin receptor expression is enhanced by T3. PMID- 16213992 TI - BMP-2 and -6 modulate porcine theca cell function alone and co-cultured with granulosa cells. AB - Bone morphogenetic proteins (BMPs) are emerging as a family of proteins crucial in the regulation of fertility and ovulation rate. We have shown that porcine theca cells express BMP receptors, however, there is a paucity of information regarding the effect(s) of BMPs on theca cell function. The purpose of this study was to investigate the effects of BMP-2 and -6 on theca cells cultured under serum-free conditions in terms of steroidogenesis, cAMP release and proliferation. The study was further extended to determine whether BMP responses in theca cells are affected by the addition of granulosa cells to the culture system. Both BMPs suppressed progesterone and androstenedione synthesis by theca cells (P < 0.05) after 144 h in culture. Oestradiol synthesis was suppressed (P < 0.05) by BMP-2, but not BMP-6, and theca cell proliferation was stimulated (P < 0.05) by BMP-6, but not BMP-2, after 144 h in culture. Both BMP-2 and -6 inhibited cAMP release (P < 0.05) by theca cells. Furthermore, progesterone and androstenedione synthesis by co-cultured theca and granulosa cells were suppressed (P < 0.05) whereas cell proliferation was stimulated (P < 0.05). These results provide strong evidence for a functional BMP system in the porcine ovary and that theca cells are responsive to BMPs in terms of steroidogenesis and proliferation. BMP-2 and -6 may have a role as luteinisation inhibitors in this polyovular species. PMID- 16213994 TI - History of the American Association of Hip and Knee Surgeons upon its 15th anniversary. PMID- 16213996 TI - Experience with an all-polyethylene total knee arthroplasty in younger, active patients with follow-up from 2 to 11 years. AB - There are few modern reports that document the results of all-polyethylene (all poly) tibial components in younger, active patients. The potential benefits of this design are the elimination of backside wear and lower implant cost than modular, metal-backed components. Nonetheless, since the mid 1980s, modular, metal-backed tibial trays have dominated the total knee arthroplasty market based on finite-element analysis studies that demonstrated superior force distribution compared with conventional all-poly components. As a result, backside wear has become an emerging problem and refocused design efforts on unitized components. Our clinical experience indicates that an all-poly tibial component fixed with cement can provide excellent performance and survivorship even in younger, active patients at intermediate follow-up. PMID- 16213997 TI - Long-term results of total knee arthroplasty in young patients with rheumatoid arthritis. AB - Forty-seven cemented total knee arthroplasties in 32 patients with rheumatoid arthritis who were 55 years or younger (average, 43) were followed until death or a minimum of 15 years. The average follow-up was 18 years. There were 6 revisions; 5 of 6 revisions had severe polyethylene wear and osteolysis. All revisions occurred after 17 years (range, 17-23). Three of these had fracture of the tibial component associated with polyethylene wear through; the remaining 2 had loose tibial and femoral components. One patient sustained a distal femoral periprosthetic fracture requiring revision of the total knee. Cemented total knee arthroplasty in the young patient with rheumatoid arthritis is reliable and durable at an average 18 years of follow-up with an estimated survivorship of 100% at 15 years and 93.7% at 20 years. PMID- 16213998 TI - Differences in patient and procedure characteristics and hospital resource use in primary and revision total joint arthroplasty: a multicenter study. AB - A multicenter retrospective cost-identification cohort study was undertaken to analyze clinical, demographic, and economic data for 4533 consecutive total hip arthroplasty (THA) and 3508 consecutive total knee arthroplasty (TKA) procedures performed during a 3-year period in 1 of 3 hospitals. Statistically significant differences were found between primary and revision procedures with respect to patient age, sex, payer type, mean total operative time, use of allograft, average length of hospital stay, discharge disposition, and hospital costs. Significant differences were also found between different types of revision total joint arthroplasty (TJA) procedures. Our findings could be used to help improve the accuracy of administrative claims data related to primary and revision TJA procedures to identify relevant differences in patient characteristics, procedure characteristics, and hospital resource use. PMID- 16213999 TI - Early results of 1000 consecutive, posterior, single-incision minimally invasive surgery total hip arthroplasties. AB - One thousand consecutive cementless total hip arthroplasties performed in 759 patients using a posterior, single-incision, minimally invasive surgical approach were followed prospectively for a minimum of 2 years. All surgeries were performed using a tapered, titanium, cementless femoral component and a press fit, ingrowth acetabular component. Mean patient age was 62.3 years. Mean incision length was 8.8 cm. Harris hip scores improved from 34 preoperatively to 92 postoperatively. Component position was considered acceptable in 95.3% of patients. Six acetabular components failed to osseointegrate, whereas no femoral component was loose. Radiolucencies were limited to the upper femur and were nonprogressive. Complications and component malpositioning were within accepted standards. This consecutive series of minimally invasive total hip arthroplasties shows that satisfactory results can be consistently achieved using this surgical technique. PMID- 16214000 TI - Outpatient total knee arthroplasty with a minimally invasive technique. AB - Fifty consecutive patients were enrolled in this prospective study. This was 37% of the 135 patients undergoing primary total knee arthroplasty (TKA) by one surgeon. The average patient age was 68 years (50-79 years). A comprehensive perioperative management pathway was developed and was implemented, which combined regional anesthesia with a minimally invasive, TKA technique in which the only incision in the capsule and extensor mechanism is a capsular incision from the joint line to the superior pole of the patella. Postoperatively, patients received oral analgesia. After specific discharge criteria were met, 48 patients (96%) chose to go home the day of surgery. No intraoperative complications occurred. There were 3 readmissions, none related to early discharge: gastrointestinal bleed at 8 days, superficial irrigation and debridement at 21 days, and a closed manipulation at 9 weeks. This study demonstrates that, in these selected patients, outpatient TKA was safe with no short-term readmission or complications related to early discharge. This comprehensive pathway may make it possible for this minimally invasive TKA to be done as an outpatient in specialized surgicenters in the future. PMID- 16214001 TI - Does shortened length of hospital stay affect total knee arthroplasty rehabilitation outcomes? AB - Rehabilitation outcomes were compared after primary total knee arthroplasty between patients who participated in a hospital joint arthroplasty program implemented to decrease length of stay and patients who did not. Once inclusion criteria were met, purposive sampling was used to select subjects for retrospective medical records review. Range of motion and Knee Society scores at preoperative and 3-, 6-, and 12-month postoperative intervals were then compared. Preoperatively, there were no significant differences between groups. Program implementation reduced length of hospital stay by a mean of 1.3 days, which resulted in a decreased range of motion at discharge. No significant differences were found between groups postoperatively at all intervals. Primary total knee arthroplasty rehabilitation outcomes were not compromised by reduced length of hospital stay. PMID- 16214002 TI - Obesity and perioperative morbidity in total hip and total knee arthroplasty patients. AB - The incidence of obesity in 1071 total hip arthroplasty (THA) patients and 1813 total knee arthroplasty (TKA) patients and its effect on perioperative morbidity were evaluated prospectively. Fifty-two percent of TKA and 36% of THA patients were obese (body mass index >or=30). The obese patients were significantly younger, with a higher proportion of obese TKA patients being women. Higher rates of diabetes and hypertension were found in obese patients. Higher postoperative infection rates were observed in patients with body mass index 35 or higher. The odds ratio was 6.7 times higher risk for infection in obese TKA patients and 4.2 times higher for obese THA patients. The increased risk of infection in obese patients undergoing total joint arthroplasty must be realized by both the patient and surgeon. PMID- 16214003 TI - Total hip arthroplasty through a minimal posterior approach using imageless computer-assisted hip navigation. AB - With decreased exposure in a minimal posterior hip incision, navigation with computer assistance provides an alternative method to accurately place the components. This study compares the results of a series of 82 navigated total hips to a retrospective cohort of 50 hips done with conventional instruments. The surgical incision split the gluteus maximus but did not extend distally into the fascia. The goal of cup placement was 40 degrees to 45 degrees of abduction (ABD) and 17 degrees to 23 degrees of flexion (FLX). Postoperative radiographs were digitized and analyzed. In the study group, 82 hips were done with computer assistance and compared with 50 done with conventional methods (manual) through the same incision. Radiographic analysis showed that there were significantly fewer cases inside the desired range of ABD and FLX in the manual group (6%) compared with the navigation group (30%), P = .001, with significant differences in the variances of ABD and FLX (P = .011 and .028). Improved accuracy of cup placement was found with increased experience in the use of navigation by the surgeon over the time of the series. The use of a computer-assisted surgery navigation system with a minimal posterior incision for a total hip arthroplasty results in significantly more reproducible acetabular component placement. PMID- 16214004 TI - Hydroxyapatite-coated femoral stem survivorship at 10 years. AB - Ninety-six patients were implanted with the Omnifit-HA femoral stem and the Omnifit-PSL porous-coated dual-radius acetabular shell (Stryker, Mahwah, NJ) and liners that were sterilized by gamma-radiation in air. The mean age at the time of surgery was 52 years (range, 27-78 years), and male/female ratio was 54:42. The mean follow-up was 10.3 years (range, 7-12 years). The survivorship of the stem and cup was 100% and 90%, respectively. Four cups were revised for aseptic loosening, and 6 liners were exchanged. The mean polyethylene wear rate was 0.24 mm/y. Lysis was noted in 23% of the cups. No patient developed femoral lysis distally. This hydroxyapatite-coated stem continues to perform well and appears to protect against the migration of wear debris along the femoral stem. PMID- 16214005 TI - The effect of total hip arthroplasty cup design on polyethylene wear rate. AB - Using 743 total hip arthroplasties that included 6 hemispheric porous-coated cup designs, this study used a multiple linear regression to identify those factors that influenced polyethylene wear rates. Wear rates for each hip were based on serial head penetration measurement made with computer-assisted techniques. Implant factors associated with an increased wear rate included terminal sterilization with a non-cross-linking chemical surface treatment, a 4-mm lateralized liner, a cobalt-chrome femoral head, and a longer shelf life for liners gamma-irradiated in air. After accounting for these implant characteristics and patient factors, wear rates among the 6 cup designs were not significantly different (P = .89). Although polyethylene wear is frequently characterized for specific implant designs, this study demonstrated that there are several common factors that influence polyethylene wear rates. PMID- 16214006 TI - Early femoral head penetration of a highly cross-linked polyethylene liner vs a conventional polyethylene liner: a case-controlled study. AB - Forty highly cross-linked, polyethylene-liner primary total hip arthroplasties were matched by sex, age, and body mass index with 40 conventional polyethylene primary total hip arthroplasties to compare femoral head penetration rates. Both inserts were 10 degrees liners manufactured by the same company and packaged in a nitrogen/vacuum atmosphere. Identical acetabular components and a 28-mm cobalt chrome femoral head were used in all cases. The mean femoral head penetration rate for the highly cross-linked and conventional polyethylene was 0.05 (range, 0.01-0.09) and 0.12 (range, 0.02-0.29) mm/y, respectively, a 58.33% reduction in femoral head penetration (P < .001). In this short-term follow-up study (mean, 47.73 months), both reduced "bedding-in" and reduced wear may contribute to the observed decrease in femoral head penetration. PMID- 16214007 TI - Impingement of acetabular cups in a hip simulator: comparison of highly cross linked and conventional polyethylene. AB - We hypothesized that the reduction in toughness that accompanies elevated levels of cross-linking in ultrahigh-molecular weight polyethylene would lead to increased wear and damage in acetabular cups under conditions of impingement with the neck of the femoral component. Test cups were exposed to 2.8, 10, and 20 Mrad of gamma irradiation and were tested in a simulator, incorporating impingement during every wear cycle. Wear behavior was compared based on wear rate, observations of damage, and the number and morphology of wear particles. The 10 Mrad cups had the lowest rate (15.2 +/- 5.1 mg/Mc), followed by the 2.8-Mrad (40.1 +/- 4.2 mg/Mc) and 20-Mrad (67.9 +/- 24.2 mg/Mc) groups. The wear damage was most severe in the 20-Mrad cups, although all 3 groups showed pitting and delaminating. Wear rates and damage of highly cross-linked polyethylene cups were significantly higher in an impingement wear test than in previous simulator studies. PMID- 16214008 TI - Cracking and impingement in ultra-high-molecular-weight polyethylene acetabular liners. AB - The purpose of this study was to determine the prevalence of crack formation in conventional ultra-high-molecular-weight polyethylene cups and its association with rim impingement, oxidation, and time in situ. One hundred twenty acetabular cups were retrieved during revision total hip arthroplasty. In 40% (48/120) of the retrieved specimens, multiple subsurface cracks of several millimeters in length were revealed by transillumination. In 5 specimens, full thickness cracks led to fragmentation of the liner before revision. Thirty-eight (32%) liners had regions of moderate to severe impingement damage to the rim; cracks were initiated at the site of impingement in all but 1 liner (P < .0001). Cracks commonly occur in conventional ultra-high-molecular-weight polyethylene liners, often after neck impingement and almost always in association with oxidation of the polymer. PMID- 16214009 TI - The long-term outcome of 755 consecutive constrained acetabular components in total hip arthroplasty examining the successes and failures. AB - Constrained acetabular components can treat or prevent instability after total hip arthroplasty (THA). We examine long-term results of 755 consecutive constrained THA in 720 patients (1986-1993; 62 primary, 59 conversion, 565 revision, 60 reimplantation, and 9 total femur). Eighty-three patients (88 THAs) were lost before 10-year follow-up, leaving 639 patients (667 THAs) available for study. Dislocation occurred in 117 hips (17.5%), in 37 (28.9%) of 128 constrained for recurrent dislocation, and 46 (28.2%) of 163 with dislocation history. Other reoperations were for aseptic loosening (51, 7.6% acetabular; 28, 4.2% stem; 16, 2.4% combined), infection (40, 6.0%), periprosthetic fracture (19, 2.8%), stem breakage (2, 0.3%), cup malposition (1, 0.1%), dissociated insert (1, 0.1%), dissociated femoral head (1, 0.1%), and impingement of 1 broken (0.1%) and 4 (0.6%) dissociated constraining rings. Although constrained acetabular components prevented recurrent dislocation in 71.1%, they should be used cautiously, with a 42.1% long-term failure rate observed in this series. Dislocation was common despite constraint with previous history as a significant risk. PMID- 16214010 TI - High failure rate of a constrained acetabular liner in revision total hip arthroplasty. AB - Fifty-five consecutive revision total hip arthroplasties (THAs) in 51 patients in which a constrained acetabular liner was used (Duraloc Constrained Liner, DePuy, Warsaw, Ind) were reviewed. In these revisions, 41 constrained liners were placed for recurrent instability and 14 for inadequate stability at the time of revision THA. The mean age of the cohort was 62 years, and 38 of the hips were in female patients (69%). At a minimum of 2 years, 9 of the 55 hips (16%) dislocated. Of these 9 dislocations, 8 occurred in patients who had undergone revision to a constrained liner for recurrent instability without femoral or acetabular component revision. Revision THA for recurrent instability by placing a constrained liner without optimizing other aspects of the reconstruction leads to a high rate of recurrent failure. PMID- 16214011 TI - Failure of femoral surface replacement for femoral head avascular necrosis. AB - The appropriate treatment for the young patient with post-collapse avascular necrosis (AVN) of the femoral head continues to be controversial. Femoral surface replacement (FSR) has emerged as a potential strategy to delay total hip arthroplasty in these patients. Between 1997 and 2003, 37 FSR procedures were performed. Success was defined as patients not requiring revision surgery or having a Harris hip pain score of 30 or greater. Failure was defined as patients having revision surgery or a Harris hip pain score of 20 or less. The overall failure rate based on Harris hip pain scores and revision surgery was 64.8% (24/37 procedures). Fifteen (40.5%) procedures required revision surgery, whereas 9 (24.3%) patients were considered failures based on their pain scores. The purpose of this study was to determine if FSR is a viable option for post collapse AVN. We conclude that hemiresurfacing for AVN is an unpredictable procedure. This procedure is no longer offered as a treatment option for post collapse AVN at our institution. PMID- 16214012 TI - Celecoxib does not affect osteointegration of cementless total hip stems. AB - Celecoxib's effect on prosthetic osteointegration in patients undergoing total hip arthroplasty was studied. In a prospective, placebo-controlled, double-blind study of 49 patients, 24 were randomized to celecoxib (200 mg/d) and 25 to placebo over 6 weeks. Bone markers, N-teleopeptide, and bone-specific alkaline phosphatase were obtained postoperatively, and at 6 and 12 weeks. No radiographic prosthetic subsidence occurred. There were no significant differences from baseline in periprosthetic bone mineral density between the celecoxib and placebo groups at 3 and 6 months. There were no significant differences in baseline BSAlkP values between the 2 groups. At 6 and 12 weeks, average BSAlkP values increased. Urinary NTx concentrations followed a skewed distribution. Normalized NTx concentrations were statistically significantly greater in the celecoxib group than the placebo group at 6 weeks but not at 12 weeks. PMID- 16214013 TI - Computer assisted navigation in total knee arthroplasty: improved coronal alignment. AB - The use of a computer navigation system is intended to optimize implant positioning. This study compares prospectively followed total knee arthroplasty (TKA) done with imageless computer-assisted navigation with a cohort of TKA done with manual instruments. Primary TKAs were performed on 147 knees, with 78 in the manual group and 69 in the navigated group. Coronal alignment was determined by postoperative full-length standing radiographs. The 2 groups had similar mean values for radiographic mechanical axis at 0.4 degrees . There was a larger variation in alignment in the manual group, with 58% of the manual group within 2 degrees of neutral compared with 78% of the navigated group (P = .008). The navigation system reproducibly resulted in a more reliable reproduction of the mechanical axis. PMID- 16214014 TI - Computer assisted navigation in total knee arthroplasty: comparison with conventional methods. AB - The success of knee arthroplasty is dependent on many factors. Postoperative extremity and component alignment are important determinants of outcome and longevity. Malalignment (>3 degrees ) results in higher failure rates. Computer assisted navigation devices were developed to improve implant positioning. This study evaluated the early outcomes of a high-volume fellowship-trained surgeon relative to component positioning and limb alignment using an image-free navigation system. The navigation group consisted of 116 consecutive patients, and the conventional group consisted of 51 consecutive patients. The postoperative mechanical axis was within 3 degrees of neutral mechanical alignment in 95% of the navigation cases vs 84% of the conventional cases (P < .02). The range of the alignment and component position measurements narrowed, and the undesired outliers decreased. Accuracy was improved with navigation. Navigation was a viable device to improve the outcome of total knee arthroplasty relative to limb and component alignment. PMID- 16214015 TI - Bacterial detection of platelets: current problems and possible resolutions. AB - The greatest transfusion-transmitted disease risk facing a transfusion recipient is that of bacterial sepsis. The prevalence of bacterial contamination in platelets and red blood cells is approximately 1 in 3,000 units transfused. The available data indicate that transfusion-associated sepsis develops after 1 in 25,000 platelet transfusions and 1 in 250,000 red blood cell transfusions. One of the most widely used strategies for decreasing bacterial sepsis risk is bacterial detection. A roundtable meeting of experts was convened during the XXVIII Annual Congress of the International Society of Blood Transfusion (Edinburgh, UK, July 2004) to provide a forum for experts to share their experiences in the routine bacterial detection of platelet products. This article summarizes the presentations, discussions, and recommendations of the panel. The data presented indicate that some of the current bacterial screening technology is useful for blocking the issuance of platelet units that contain relatively high levels of contaminating bacteria. Platelet units are usually released based on a test negative status, which often become test-positive only upon longer storage. These data thus suggest that bacterial screening may not prevent all transfusion transmitted bacterial infections. Two transfusion-transmitted case reports further highlighted the limitation of the routine bacterial screening of platelet products. It was felt that newer technologies, such as pathogen inactivation, may represent a more reliable process, with a higher level of safety. The panel thus recommended that the Transfusion Medicine community may need to change its thinking (paradigm) about bacterial detection, toward the possibility of the pathogen inactivation of blood products, to deal with the bacterial contamination issue. It was suggested, where permitted by regulatory agencies, that blood centers should consider adopting first-generation pathogen inactivation systems as a more effective approach to reducing the risk of transfusion-associated sepsis than some of the approaches currently available. PMID- 16214016 TI - Risk management in transfusion after the HIV blood contamination crisis in France: the impact of the precautionary principle. AB - The importance of the precautionary principle in public health was highlighted in France after the HIV contamination of blood products used for transfusion. However, the definition of this principle, its objectives, the way in which it should be applied, and its consequences had not been considered previously. The question as to whether the application of the precautionary principle is appropriate remains unanswered. The aim of this study was to analyze the interpretations of the application of the precautionary principle to determine its consequences in terms of risk management and patient rights. This was accomplished by interviewing persons involved in transfusion medicine. We conducted 33 interviews and describe the issues enunciated for and against the application of the precautionary principle. The precautionary principle concept was confusing to the respondents. A major issue emerging from the interviews was that the precautionary principle was perceived more as a means of protecting the decision maker than as a means of protecting the patient. Taken to its extreme, the use of the precautionary principle could prejudice sound medical decision making. However, it was felt that it also can lead to the introduction of measures that update and gradually reduce risks associated with transfusion. PMID- 16214018 TI - Transfusion medicine in the era of genomics and proteomics. AB - Viewing recent trends in transfusion medicine (TM), the authors make predictions about possible future developments within this specialty including greater cost effectiveness and blood safety resulting from increased automation; techniques in genetics replacing serological typing in many standard assays; and TM service playing a major R&D role together with clinical services in the emerging cell based therapeutics. To achieve this, the TM laboratory of the future will need to have available extensive skills in immunogenetics and database expertise; emerging techniques in genomics and proteomics will need to be integrated with classic immunohematology approaches; and collaborative networks of TM laboratories will need to raise their profiles as a competent partner in the ongoing clinical biotechnology revolution. Blood product safety is profiled to highlight some of these developments. Until recently, avoiding pathogen transmission has focused primarily on excluding at-risk donors and testing donor blood for pathogen markers. Newer trends in pathogen-inactivation procedures could alter the protein composition of the blood product, potentially causing unintended immune reactions that could outweigh their benefits in further reducing a very low current risk of pathogen transmission. By combining proteomics and immunohematology, those manufacturing processes least likely to generate posttranslational protein modifications will need to be identified. PMID- 16214017 TI - The putative mechanism of the immunomodulating effect of HLA-DR shared allogeneic blood transfusions on the alloimmune response. AB - Pretransplant blood transfusions have shown to improve organ allograft survival. However, the immunologic mechanism leading to this beneficial effect in clinical transplantation is still not clear. The observation that blood transfusions sharing an HLA-DR antigen with the recipient are more effective than HLA mismatched transfusions has led to the hypothesis that CD4(+) regulatory T (Treg) cells that recognize a foreign peptide in the context of the shared HLA-DR molecule play an important role in down-regulation of the immune response toward the graft. Available experimental evidence supports this hypothesis. Furthermore, these CD4(+) Treg cells are able to modulate antigen-presenting cells, which in their turn will induce Treg cells. As long as clinical transplantation tolerance by blood transfusions is not a reality, further studies on the mechanisms of the beneficial effect of pretransplant allogeneic blood transfusions are needed to obtain an effective protocol for the induction of clinically relevant Treg cells. PMID- 16214019 TI - Blood group biochemistry: a canadian blood services research and development symposium. PMID- 16214022 TI - Improved psychomotor performance in aged mice fed diet high in antioxidants is associated with reduced ex vivo brain interleukin-6 production. AB - Psychomotor performance is decreased in the aged. This study investigated the relationship between brain oxidative stress, interleukin-6 (IL-6) production by brain tissue ex vivo and psychomotor deficits during aging, and the effects of feeding an antioxidant-rich diet on ex vivo brain IL-6 production and motor function in aged mice. Male BALBc mice reared in SPF conditions and ranging in age from 3 to 24 months were studied. There was a precipitous decline in motor function after 12 months of age and an increase in brain lipid peroxidation and IL-6 production by coronal brain slices ex vivo. In another study, 12-month-old mice were fed diets formulated to provide a disparate range of antioxidants. At 18 months of age psychomotor coordination, motor learning, and ex vivo brain IL-6 production were evaluated. Mice fed an antioxidant-rich diet had improved psychomotor coordination compared to mice fed diet adequate or low in antioxidants. When mice were tested on successive days, only those fed adequate and high antioxidants exhibited motor learning. Analysis of IL-6 production by coronal brain slices indicated that as dietary antioxidants increased, IL-6 production decreased. Collectively, these data indicate that antioxidants improve psychomotor performance in aged mice, and suggest antioxidants may be useful for reducing brain IL-6 production, which has been shown to increase in aged mice. PMID- 16214023 TI - Interleukin-6 levels fluctuate with the light-dark cycle in the brain and peripheral tissues in rats. AB - Interleukin-6 (IL-6) has been implicated in excessive daytime sleepiness (EDS) in humans, and exogenous IL-6 also induces sleep alterations both in humans and rats. The IL-6 levels in human blood vary with the light-dark cycle with IL-6 levels being high during the dark period and low during the light period, whereas in the pituitary of rats the IL-6 levels are elevated during the light period compared to the dark period. However, it is unknown whether IL-6 in the brain is affected by the light-dark cycle. We hypothesized that IL-6 levels in the brain are regulated by the light-dark cycles and are elevated during the period that is predominantly occupied by sleep. To test this hypothesis, we measured IL-6 levels in the brain, blood, and adipose tissue of rats across light-dark cycle every 4 h. IL-6 levels were significantly higher during the light period than during the dark period in the cortex, hippocampus and hypothalamus. In the brainstem, IL-6 levels did not significantly vary across the light-dark cycles. IL-6 levels in the blood and adipose tissues were also significantly higher during the light period than during the dark period. IL-6 levels were positively correlated between the blood and adipose tissue, between hypothalamus and blood, and between the hypothalamus and hippocampus. These observations suggest that IL-6 levels vary across the light-dark cycle among different tissues and that IL-6 levels are elevated both centrally and peripherally during the period predominantly occupied by sleep but decreased during the period that primarily consists of wakefulness. PMID- 16214024 TI - Time-dependent behavioral, neurochemical, and immune consequences of repeated experiences of social defeat stress in male mice and the ameliorative effects of fluoxetine. AB - This study attempted to determine whether differing numbers of days of repeated defeat experience altered behavior, immune measures, and neuroendocrine mediators in mice. OF1 male mice were socially stressed by repeated experiences of defeat in a sensorial contact model. Subjects exposed to nine defeats showed more stretch-attend postures and fewer active defense elements than counterparts exposed to 23 defeats. Submissive subjects with nine experiences of defeat also had a lower splenocyte proliferative response than unmanipulated controls. The proliferation index progressively increased but at a higher rate in manipulated controls than in socially stressed subjects, resulting in a significant immunosuppressive effect after 23 days of exposure to social stressors. Nine days of such exposure resulted in higher hypothalamic ratios of serotonin and dopamine to their major metabolites than in unmanipulated or manipulated controls and subjects socially stressed for 23 days. The data generally indicate that the acute social stressors (such as nine defeats) produce a profile of behavioral and physiological variables characteristic of a state of anxiety. The proliferation index was also lower after 52 days of social stress than in manipulated controls. Fluoxetine treatment appeared to have an anxiolytic effect, reducing immobility, and even seemed to protect subjects from the immune impairment and endocrine alteration caused by social stressors. The results generally provide clues that improve our knowledge of the consequences of social stressors and their possible treatment. PMID- 16214025 TI - Psychological stress activates interleukin-1beta gene expression in human mononuclear cells. AB - The pathophysiological mechanisms underlying the association between psychological stress and cardiovascular disease are unclear. Interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) are inflammatory cytokines playing a pivotal role in atherosclerosis. IL-1beta activates IL-6, and both cytokines are produced by peripheral blood mononuclear cells. One mechanism through which stress could promote atherosclerosis is by regulating mononuclear cell cytokine gene expression. We studied cardiovascular and cytokine responses in 32 healthy men participating in two 5-min mental tasks and in 10 controls. Blood pressure and heart rate, assessed using a Portapres-2, increased significantly following tasks in all participants. Plasma IL-6 levels, determined by ELISA, also increased following tasks, with maximum levels detected 2h post-stress. Quantitative RT-PCR analysis showed that mononuclear cell IL-1beta gene expression rose significantly at 30 min post-stress and remained elevated at 75 and 120 min. Increases in IL 1beta gene expression correlated positively with plasma IL-6 responses, cardiovascular responses, subjective stress ratings, and anxiety symptoms. No changes were detected in controls. Stress-induced activation of mononuclear IL 1beta is a novel mechanism potentially linking stress and heart disease. This mechanism could also play a role in other inflammatory diseases exacerbated by stress. PMID- 16214026 TI - Self-rated health and vital exhaustion, but not depression, is related to inflammation in women with coronary heart disease. AB - Poor subjective well-being has been associated with increased coronary heart disease (CHD) morbidity and mortality in population-based studies and with adverse outcomes in existing CHD. Little is known about the mechanisms responsible for this association, but immune activity appears to be a potential pathway. Despite the growing evidence linking immune activity to subjective feelings, very few studies have examined patients with CHD, and the results are conflicting. We examined consecutive women patients hospitalized for acute myocardial infarction, and/or underwent percutaneous transluminal coronary angioplasty or coronary artery bypass grafting. We assessed depression, vital exhaustion, and self-rated health by questionnaires. Circulating levels of high sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), and interleukin-1 receptor antagonist (IL-1ra) concentrations were determined. After controlling for potential confounding factors there was a significant positive correlation between IL-6 levels and vital exhaustion and poor self-rated health. The association between hsCRP and vital exhaustion and self-rated health was borderline significant. In contrast, the correlations between psychological factors and IL-1ra levels were weak and non-significant, as were the correlations between inflammatory markers and depression. Similar relationships between the inflammatory markers and the measures of psychological well-being were obtained when the latter ones were categorized into tertiles. In conclusion, inflammatory activity, assessed by IL-6 and hsCRP levels, was associated with vital exhaustion and self-rated health in CHD women. These findings may provide further evidence for a possible psychoneuroimmune link between subjective well-being and CHD. Our observations also raise the possibility that a cytokine-induced sickness response in CHD may be better represented by constructs of vital exhaustion and self-rated health than of depression. PMID- 16214027 TI - The effects of lipopolysaccharide and lithium chloride on the ingestion of a bitter-sweet taste: comparing intake and palatability. AB - Activation of the immune system with lipopolysaccharide (LPS) has been shown to result in decreased consumption of normally preferred substances while at the same time not affecting palatability. The present study examined the effects LPS administration on both intake and palatability of a relatively unpalatable bitter sweet taste. Bitter is thought to signal a danger cue to an animal representing a potential toxin-containing food. Using a one-bottle consumption test, voluntary intake of a sucrose-quinine (0.15 M sucrose + 0.00015 M quinine; S-Q) solution was assessed in rats on two conditioning days (days 1 and 4) after a systemic injection with LPS, LiCl, or NaCl. On the test day (day 7), rats were given 1h access to the same solution in the absence of any injection. In a separate experiment, rats fitted with intraoral cannulae received similar testing schedules, however, the solution was delivered intraorally, activating only the consummatory responses of the animal. During conditioning, rats received 5 brief (1 min) intraoral infusions of the taste across a 1h period following injections of LPS, LiCl or NaCl. Individual taste reactivity responses were recorded and analyzed. Both LPS and LiCl resulted in decreased consumption of the unpalatable taste relative to controls on the test day, suggesting typical conditioned taste avoidance. When the consummatory responses were examined, LPS-treatment produced an increase in active oral rejection relative to NaCl- and LiCl-treated groups on both conditioning days. The present study demonstrates that although both LPS- and LiCl-treatment result in similar conditioned avoidance using an intake measure, they do not elicit similar patterns of taste reactivity responding to intraoral infusions of the bitter-sweet taste. Furthermore, the present results suggest that immune activation with LPS-treatment results in increased rejection of a mildly aversive stimulus and supports the hypothesis that reorganization of behavioral priorities occurs during bacteria-induced sickness. PMID- 16214028 TI - A confocal microscopy study of the very early cellular response to oxidative stress induced by zinc phthalocyanine sensitization. AB - Oxidative stress has been involved in several biological and pathological processes. Reactive oxygen species have been shown to play both beneficial and deleterious roles. The present work contributes to the understanding of the very early events of cellular response to oxidative stress. Oxidative stress was produced intracellularly by light activation of zinc phthalocyanine (ZnPc) at a light dose that did not lead to apoptosis or necrosis. Phthalocyanine was photoactivated using the 647-nm laser line of a confocal microscope through the objective lens causing oxidative stress and allowing observation of the evoked phenomena at the single cell level and in real time. Mitochondria membrane potential (DeltaPsi(m)), intracellular pH, calcium concentration, and generation of reactive oxygen species (ROS) were recorded using specific vital fluorescent probes and quantified by image processing and analysis. Subcellular localization of ZnPc was also studied in order to determine the primary and intermediate ROS target. PMID- 16214029 TI - Induction of apoptosis by adenovirus-mediated manganese superoxide dismutase overexpression in SV-40-transformed human fibroblasts. AB - Although mitochondrial reactive oxygen species (ROS) have been implicated both as an initiator and as an effector of apoptosis, the exact role of mitochondrial ROS has been difficult to establish due to the lack of an appropriate experimental system where ROS could be specifically generated from mitochondria and subsequent effects on cells analyzed. In this study, a manganese superoxide dismutase (MnSOD) activity-mediated apoptosis model was established and characterized. It was shown that despite early increases in the steady-state levels of ROS upon MnSOD overexpression, cellular oxidative damage was decreased significantly at later time points. Alterations in levels of peroxiredoxin (Prxn1) protein preceded the onset of apoptosis after MnSOD overexpression. A time course study demonstrated that increases in MnSOD activity prior to the onset of apoptosis correlated with alterations in the levels of nitration of tyrosine residue(s) of MnSOD protein. A direct correlation between MnSOD activity and the degree of apoptosis was demonstrated using a mutant MnSOD with decreased activity. The current study supports a causative role of mitochondrial ROS leading to the onset of apoptosis. The MnSOD activity-mediated apoptosis model described here could be further utilized to study mitochondrial apoptotic pathways. PMID- 16214030 TI - The proteasome inhibitor MG132 protects against acute pancreatitis. AB - The cell-permeant MG132 tripeptide (Z-Leu-Leu-Leu-aldehyde) is a peptide aldehyde proteasome inhibitor that also inhibits other proteases, including calpains and cathepsins. By blocking the proteasome, this tripeptide has been shown to induce the expression of cell-protective heat shock proteins (HSPs) in vitro. Effects of MG132 were studied in an in vivo model of acute pancreatitis. Pancreatitis was induced in male Wistar rats by injecting 2 x 100 microug/kg cholecystokinin octapeptide intraperitoneally (ip) at an interval of 1 h. Pretreating the animals with 10 mg/kg MG132 ip before the induction of pancreatitis significantly inhibited IkappaB degradation and subsequent activation of nuclear factor-kappaB (NF-kappaB). MG132 also increased HSP72 expression. Induction of HSP72 and inhibition of NF-kappaB improved parameters of acute pancreatitis. Thus MG132 significantly decreased serum amylase, pancreatic weight/body weight ratio, pancreatic myeloperoxidase activity, proinflammatory cytokine concentrations, and the expression of pancreatitis-associated protein. Parameters of oxidative stress (GSH, MDA, SOD, etc.) were improved in both the serum and the pancreas. Histopathological examinations revealed that pancreatic specimens of animals pretreated with the peptide demonstrated milder edema, cellular damage, and inflammatory activity. Our findings show that simultaneous inhibition of calpains, cathepsins, and the proteasome with MG132 prevents the onset of acute pancreatitis. PMID- 16214031 TI - Oxysterol-induced up-regulation of MCP-1 expression and synthesis in macrophage cells. AB - To investigate the proinflammatory potential of cholesterol and cholesterol oxidation products (oxysterols), which are present in oxidized low-density lipoproteins, foam cells, and fibrotic plaque, we used an in vitro model mimicking the challenge of macrophage cells by the cholesterol accumulating within the central core of atheroma. A biologically representative oxysterol mixture was shown to be potentially able to sustain a chronic inflammatory process within the vascular wall by up-regulating the expression of defined proinflammatory genes. In particular, expression and synthesis of the major chemokine for monocytes/macrophages, namely monocyte chemotactic protein-1 (MCP 1), were consistently increased when cells of the macrophage lineage (U937 cell line) were incubated with this mixture. On the contrary, an identical concentration of unoxidized cholesterol in no case modified expression or synthesis of the chemokine. Up-regulated expression and synthesis of MCP-1 by the oxysterol mixture was clearly dependent on a net increment of phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor kappaB (NF kappaB) nuclear binding. The results indicate that cholesterol may contribute to the progression of atherosclerotic lesions by strongly up-regulating crucial proinflammatory factors like MCP-1, but only after having been oxidized to oxysterols. PMID- 16214032 TI - Induction of endothelial cell apoptosis by lipid hydroperoxide-derived bifunctional electrophiles. AB - Endothelial dysfunction is considered to be the earliest event in atherogenesis. Oxidative stress, inflammation, and apoptosis play critical roles in its progression and onset. Lipid peroxidation, which occurs during oxidative stress, results in the formation of lipid hydroperoxide-derived bifunctional electrophiles such as 4-hydroxy-2(E)-nonenal that induce apoptosis. In this study, recently identified lipid hydroperoxide-derived bifunctional electrophiles 4-oxo-2(E)-nonenal (ONE; 5-30 microm) and 4,5-epoxy-2(E)-decenal (EDE; 10-20 microM) were shown to cause a dose- and time-dependent apoptosis in EA.hy 926 endothelial cells. This was manifest by morphological changes, caspase-3 activation, and poly(ADP-ribose) polymerase cleavage. Bifunctional electrophiles caused cytochrome c release from mitochondria into the cytosol, implicating a mitochondrial pathway of apoptosis in the endothelial cells. The novel carboxylate-containing lipid hydroperoxide-derived bifunctional electrophile 9,12 dioxo-10(E)-dodecenoic acid was inactive because it could not translocate across the plasma membrane. However, its less polar methyl ester derivative (2-10 microM) was the most potent inducer of apoptosis of any bifunctional electrophile that has been tested. An acute decrease in intracellular glutathione (GSH) preceded the onset of apoptosis in bifunctional electrophile-treated cells. The ability of ONE and EDE to deplete GSH was directly correlated with their predicted reactivity toward nucleophilic amino acids. Liquid chromatography/mass spectrometry methodology was developed in order to examine the intracellular and extracellular concentrations of bifunctional electrophile-derived GSH adducts. Relative intracellular/extracellular ratios of the GSH adducts were identical with the rank order of potency for inducing caspase 3 activation. This suggests that there may be a role for the bifunctional electrophile-derived GSH adducts in the apoptotic response. N-Acetylcysteine rescued bifunctional electrophile treated cells from apoptosis, whereas the GSH biosynthesis inhibitor d,l buthionine-(R,S)-sulfoximine sensitized the cells to apoptosis. These data suggest that lipid hydroperoxide-derived bifunctional electrophiles may play an important role in cardiovascular pathology through their ability to induce endothelial cell apoptosis. PMID- 16214033 TI - Benzo[a]pyrene and its metabolites combined with ultraviolet A synergistically induce 8-hydroxy-2'-deoxyguanosine via reactive oxygen species. AB - We previously reported that benzo[a]pyrene (BaP) and UVA radiation synergistically induced oxidative DNA damage via 8-hydroxy-2'-deoxyguanosine (8 OHdG) formation in vitro. The present study shows that microsomal BaP metabolites and UVA radiation potently enhance 8-OHdG formation in calf thymus DNA about 3 fold over the parent compound BaP. Utilization of various reactive oxygen species scavengers revealed that singlet oxygen and superoxide radical anion were involved in the 8-OHdG formation induced by microsomal BaP metabolites and UVA. Two specific BaP metabolites, benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide (+/-) (anti) (BPDE) and BaP-7,8-dione, were further tested for synergism with UVA. BaP-7,8-dione showed an effect on 8-OHdG formation induced by UVA radiation that was similar to that of the parent BaP, whereas BPDE exhibited significantly higher induction of 8-OHdG than BaP. At as low as 0.5 microM, BPDE plus UVA radiation substantially increased 8-OHdG levels about 25-fold over the parent BaP. BPDE increased the formation of 8-OHdG levels in both BPDE concentration- and UVA dose-dependent manners. Additionally, singlet oxygen was found to play a major role in 8-OHdG induction by BPDE and UVA. These results suggest that BaP metabolites such as BPDE synergize with UVA radiation to produce ROS, which in turn induce DNA damage. PMID- 16214034 TI - Oxypurinol improves coronary and peripheral endothelial function in patients with coronary artery disease. AB - Coronary endothelial dysfunction is a powerful prognostic marker in patients with coronary artery disease (CAD) that is centrally related to oxidative inhibition of nitric oxide (NO)-dependent vascular cell signaling. Xanthine oxidase (XO), which both binds to and is expressed by endothelial cells, generates superoxide and hydrogen peroxide upon oxidation of purines. Whether inhibition of xanthine oxidase activity results in improved coronary vasomotor function in patients with CAD, however, remains unknown. We assessed coronary and peripheral (brachial artery) endothelial function in 18 patients (pts; 65+/-8 years, 86% male) with angiographically documented CAD, preserved left ventricular function, and non elevated uric acid levels (233+/-10 microM). Patients received incremental doses of intracoronary acetylcholine (ACh; 10(-7) to 10(-5) microM), and minimal lumen diameter (MLD) and coronary blood flow (CBF) were assessed before and after intravenous administration of oxypurinol (200 mg). Oxypurinol inhibited plasma XO activity 63% (0.051+/- 0.001 vs 0.019+/- 0.005 microU/mg protein; p<0.01). In pts who displayed endothelial dysfunction as evidenced by coronary vasoconstriction in response to ACh (n=13), oxypurinol markedly attenuated ACh-induced vasoconstriction (-23+/- 4 vs -15+/- 4% at ACh 10(-5) microM, p<0.05) and significantly increased CBF (16+/-17 vs 62+/-18% at ACh 10(-5) microM, p<0.05), whereas in patients with preserved coronary endothelial function, oxypurinol had no effect on ACh-dependent changes in MLD (+2.8+/- 4.2 vs 5.2+/- 0.7%, p>0.05) or CBF (135+/-75 vs 154+/-61%, p>0.05). Flow-mediated dilation of the brachial artery, assessed in eight consecutive patients, increased from 5.1+/-1.5 before to 7.6+/-1.5% after oxypurinol administration (p < 0.05). Oxypurinol inhibition of XO improves coronary vascular endothelial dysfunction, a hallmark of patients with CAD. These observations reveal that XO-derived reactive oxygen species significantly contribute to impaired coronary NO bioavailability in CAD and that XO inhibition represents an additional treatment concept for inflammatory vascular diseases that deserves further investigation. PMID- 16214035 TI - Selective nitros(yl)ation induced in vivo by a nitric oxide-donating cyclooxygenase-2 inhibitor: a NObonomic analysis. AB - Nitric oxide (NO) enhances anti-inflammatory drug action. Through a metabonomics approach termed "NObonomics," the effects of a prototypic NO donor (organic nitrate)-cyclooxygenase-2 inhibitor hybrid (NO-coxib), NMI-1093, on the NO metabolite status of the circulation and major organs have been profiled in vivo in the rat. An oral anti-inflammatory NMI-1093 bolus elicited acute tissue-, time , and dose-dependent changes in oxidative and nitroso/nitrosyl NO metabolites. Gastric N-nitrosation and hepatic S-nitrosation and heme nitrosylation emerged as sensitive indices of this NO-coxib's metabolism. Acute NMI-1093-induced nitros(yl)ation correlated positively as a function of nitrate plus nitrite formation across all organs examined, suggesting a unifying in vivo mechanism consequent to NMI-1093 biotransformation that links oxidative and nitros(yl)ative routes of NO chemical biology and thereby may support downstream NO signaling. NMI-1093 depressed erythrocyte nitros(yl)ation, likely by inhibiting cellular carbonic anhydrase and shifting the intracellular balance between nitrogen oxides and carbonates. Glutathione-S-transferase or cytochrome P450 inhibitors also attenuated NMI-1093's NO metabolism in a compartment-selective fashion. Although not itself a NO donor, the des-nitro coxib analog of NMI-1093 influenced basal NO metabolite profiles, implicating a cyclooxygenase-NO synthase interaction in physiological NO regulation. By detailing the global NO metrics of a unique coxib bearing a popular NO-donor pharmacophore (i.e., a nitrate moiety) and defining some critical mechanistic determinants, this study demonstrates how NObonomics can serve as valuable tool in helping elucidate NO systems biology and the effect of NO-donor and non-NO-donating therapeutics thereon. PMID- 16214036 TI - Protein oxidation and degradation during postmitotic senescence. AB - Oxidized and cross-linked proteinacious materials (lipofuscin, age pigments, ceroid, etc.) have long been known to accumulate in aging and in age-related diseases, and some studies have suggested that age-dependent inhibition of the proteasome and/or lysosomal proteases may contribute to this phenomenon. Cell culture studies trying to model these aging effects have almost all been performed with proliferating (divisionally competent) cell lines. There is little information on nondividing (postmitotic) cells; yet age-related accumulation of oxidized and cross-linked protein aggregates is most marked in postmitotic tissues such as brain, heart, and skeletal muscles. The present investigation was undertaken to test whether oxidized and cross-linked proteins generally accumulate in nondividing, IMR-90 and MRC-5, human cell lines, and whether such accumulation is associated with diminished proteolytic capacities. Since both protein oxidation and declining proteolytic activities might play major roles in the age-associated accumulation of intracellular oxidized materials, we tested for protein carbonyl formation, proteasomal activities, and lysosomal cathepsin activities. For these studies, confluent, postmitotic IMR-90 and MRC-5 fibroblasts (at various population doubling levels) were cultured under hyperoxic conditions to facilitate age-related oxidative senescence. Our results reveal marked decreases in the activity of both the proteasomal system and the lysosomal proteases during senescence of nondividing fibroblasts, but the peptidyl-glutamyl hydrolyzing activity of the proteasome was particularly inhibited. This decline in proteolytic capacity was accompanied by an increased accumulation of oxidized proteins. PMID- 16214037 TI - A new sensitive assay reveals that hemoglobin is oxidatively modified in vivo. AB - Free radical formation in heme proteins is recognised as a factor in mediating the toxicity of peroxides in oxidative stress. As well as initiating free radical damage, heme proteins damage themselves. Under extreme conditions, where oxidative stress and low pH coincide (e.g., myoglobin in the kidney following rhabdomyolysis and hemoglobin in the CSF subsequent to subarachnoid hemorrhage), peroxide can induce covalent heme to protein cross-linking. In this paper we show that, even at neutral pH, the heme in hemoglobin is covalently modified by oxidation. The product, which we term OxHm, is a "green heme" iron chlorin with a distinct optical spectrum. OxHm formation can be quantitatively prevented by reductants of ferryl iron, e.g., ascorbate. We have developed a simple, robust, and reproducible HPLC assay to study the extent of OxHm formation in the red cell in vivo. We show that hemoglobin is oxidatively damaged even in normal blood; approximately 1 in 2,000 heme groups exist as OxHm in the steady state. We used a simple model (physical exercise) to demonstrate that OxHm increases significantly during acute oxidative stress. The exercise-induced increase is short-lived, suggesting the existence of an active mechanism for repairing or removing the damaged heme proteins. PMID- 16214038 TI - Role of endothelial nitric oxide in microvascular oxygen delivery and consumption. AB - Nitric oxide (NO) is an important signaling molecule modulating diverse processes such as vasodilation, neurotransmission, long-term potentiation, and immune responses. The endothelium contributes a significant fraction of NO from endothelial NO synthase (eNOS). The objective of this work was to analyze the role of eNOS in the modulation of oxygen supply to the tissues and in adaptation to maintain oxygenation uncompromised. Oxygen delivery and consumption were measured in the microcirculation of homozygous mutant endothelial nitric oxide synthase-deficient (eNOS(-/-)) and wild-type mice. Animals were implanted with a dorsal window chamber, allowing us to assess the intact microvascular system. Hemodynamics and oxygen tension were assessed in the microcirculation of conscious animals. The eNOS(-/-) mice had significantly higher blood pressure and lower heart rate (146 +/- 8 mm Hg, 401 +/- 17 bpm) than wild type (127 +/- 6 mm Hg, 428 +/- 20 bpm). Microvascular hemodynamic parameters were not significantly different between groups. The eNOS(-/-) animals delivered less oxygen to the microcirculation and released more oxygen to the tissue; both differences were statistically significant compared to wild type. The arteriolar vessel wall oxygen gradient, a measure of vascular smooth muscle cells and endothelial cell wall oxygen consumption, was significantly lower for eNOS(-/-) than for wild type, suggesting that the inhibition of eNOS is an antianoxia (oxygen sparing) mechanism. Finally, the findings of the study support the argument that NO availability limits oxygen consumption by the tissue. PMID- 16214039 TI - Exogenous superoxide mediates pro-oxidative, proinflammatory, and procoagulatory changes in primary endothelial cell cultures. AB - Endothelial dysfunction/activation underlies the development of long-term cardiovascular complications and atherosclerosis. The aim of this study was to examine a direct role for exogenous sublethal flux of superoxide on endothelial cell dysfunction. Human umbilical vein endothelial cells (HUVEC) were exposed to superoxide generated by 0.1 mM xanthine and 4 mU/ml xanthine oxidase for 15 min and essential endothelial functions were examined. Superoxide dismutase and/or catalase was used as scavenger for O(2)(-)/H(2)O(2) to determine the key culprit. HUVEC detachment was determined by neutral red uptake and apoptosis by annexin V binding. Inflammation was estimated by IL-8 mRNA expression and cellular adhesion molecules (CAM). eNOS and iNOS message and eNOS protein served as an indirect measure for NO. Procoagulable state was evaluated by estimating the intracellular tissue factor. Activation of endothelial NADPH oxidase was determined by lucigenin chemiluminescence. Sublethal superoxide dose evoked: (1) proinflammatory state manifested by increased IL-8 mRNA expression and CAM on the endothelial surface, (2) HUVEC apoptosis and activated endothelial NADPH oxidase, (3) increase in intracellular tissue factor, and (4) decrease in eNOS mRNA and protein and up-regulation of iNOS mRNA. We conclude that extracellular low flux of superoxide exhibits pleiotropic characteristics, triggering activation/dysfunction of endothelial cells. PMID- 16214040 TI - Fat intake reverses the beneficial effects of low caloric intake on skeletal muscle mitochondrial H(2)O(2) production. AB - Food restriction is the most effective modulator of oxidative stress and it is believed that a reduction in caloric intake per se is responsible for the reduced generation of reactive oxygen species (ROS) by mitochondria. Hydrogen peroxide (H(2)O(2)) generation and oxygen consumption (O(2)) by skeletal muscle mitochondria were determined in a peculiar strain of rats (Lou/C) characterized by a self-low-caloric intake and a dietary preference for fat. These rats were fed either with a standard high-carbohydrate (HC) or a high-fat (HF) diet and the results were compared to those measured in Wistar rats fed a HC diet. H(2)O(2) production was significantly reduced in Lou/C rats fed a HC diet; this effect was not due to a lower O(2) consumption but rather to a decrease in rotenone sensitive NADH-ubiquinone oxidoreductase activity and increased expression of uncoupling proteins 2 and 3. The reduced H(2)O(2) generation displayed by Lou/C rats was accompanied by a significant inhibition of permeability transition pore (PTP) opening. H(2)O(2) production was restored and PTP inhibition was relieved when Lou/C rats were allowed to eat a HF diet, suggesting that the reduced oxidative stress provided by low caloric intake is lost when fat proportion in the diet is increased. PMID- 16214042 TI - Human ubiquitin specific protease 31 is a deubiquitinating enzyme implicated in activation of nuclear factor-kappaB. AB - TRAF2 mediates activation of the transcription factors NF-kappaB and AP1 by TNF. A yeast two-hybrid screen of a human cDNA library identified a ubiquitin specific protease homologue (USP31) as a TRAF2-interacting protein. Two cDNAs encoding for USP31 were identified. One cDNA encodes a 1035-amino acid long isoform of USP31 (USP31, long isoform) and the other a 485-amino acid long isoform of USP31 (USP31S1, short isoform). USP31 and USP31S1 share a common amino terminal region with homology to the catalytic region of known deubiquitinating enzymes. Enzymatic assays demonstrated that USP31 but not USP31S1 possess deubiquitinating activity. Furthermore, it was shown that USP31 has a higher activity towards lysine-63-linked as compared to lysine-48-linked polyubiquitin chains. Overexpression of USP31 in HEK 293T cells inhibited TNFalpha, CD40, LMP1, TRAF2, TRAF6 and IKKbeta-mediated NF-kappaB activation, but did not inhibit Smad mediated transcription activation. In addition, both USP31 isoforms interact with p65/RelA. Our data support a role for USP31 in the regulation of NF-kappaB activation by members of the TNF receptor superfamily. PMID- 16214041 TI - HMG-CoA reductase inhibitors upregulate heme oxygenase-1 expression in murine RAW264.7 macrophages via ERK, p38 MAPK and protein kinase G pathways. AB - Heme oxygenase-1 (HO-1) is the rate-limiting enzyme in heme catabolism, which confers cytoprotection against oxidative injury and provides a vital function in maintaining tissue homeostasis. HMG-CoA reductase inhibitors (statins) possess several anti-inflammatory mechanisms and may be beneficial in the treatment of inflammatory diseases. Our previous study has shown that statins can inhibit iNOS gene expression in murine RAW264.7 macrophages. In this study, we showed that lovastatin, fluvastatin, atorvastatin, simvastatin, mevastatin and pravastatin are able to upregulate the mRNA expression of HO-1 gene. This effect of lovastatin was attenuated by farnesyl pyrophosphate (FPP), geranylgeranyl pyrophosphate (GGPP), a protein kinase G (PKG) inhibitor (KT5823), a soluble guanylyl cyclase inhibitor (ODQ), a p38 MAPK inhibitor (SB203580), and MEK inhibitors (U0126 and PD98059), but not by inhibitors of protein kinase C (PKC), protein kinase A (PKA), c-jun N-terminal kinase (JNK) and Rho kinase. Consistent with this notion, our previous study has reported the ability of statins to activate ERK and p38 MAPK in RAW264.7 macrophages. Here we further found the participation of cyclic guanosine monophosphate (cGMP)/PKG pathway for ERK activation in cells stimulated with statin and the ability of statin to induce AP 1 activity, which is an essential transcription factor in the regulation of HO-1 gene expression. In addition, a Ras inhibitor (manumycin A) treatment also caused a marked induction of HO-1 mRNA followed by a corresponding increase in HO-1 protein; instead, inhibition of Rho activity by toxin B only led to a transient and weak induction of HO-1. The involvement of signal pathways in manumycin A induced HO-1 gene expression was associated with p38 MAPK, JNK and ERK activation. Taken together, these results demonstrate for the first time that statins might activate PKG to elicit activations of ERK and p38 MAPK pathways and finally induce HO-1 gene expression, which provides a novel anti-inflammatory mechanism in the therapeutic validity. PMID- 16214043 TI - The difficulties industry is facing with investigators. AB - The number of new agents being developed for the treatment of cancer has, over the past 10 years, increased dramatically which has resulted in increased interactions between the pharmaceutical industry that discover and develop most new agents and investigators in academic institutions, hospitals and office practices. This close interaction has inevitably led to a number of issues being identified on both sides and this paper will attempt to identify some of these and propose solutions. PMID- 16214044 TI - Disclosure of competing financial interests and role of sponsors in phase III cancer trials. AB - Financial relationships between industry, researchers and academic institutions are becoming increasingly complex, raising concern about sponsors' involvement in the conduct of biomedical research. A review of published randomised trials (RCTs) in cancer research was performed to assess adherence to the 1997 disclosure requirements and to document the nature of the disclosed interests. Source(s) of study support, author-sponsor relationships and the role of the study sponsor were assessed for all RCTs published between 1999 and 2003 in 12 international journals. A total of 655 cancer RCTs were identified. Of these, 516 (78.8%) disclosed the source of sponsorship. The nature of the relationship between the authors and the study sponsor was included in 219 of the 227 industry sponsored studies. The most commonly cited relationships were (131 studies had multiple relations): grants (93.6%); employment (39.2%); consultant/honorarium (12.7%) and stock ownership and participation in a speaker's bureau (12, 5.5% each). Only 41 (18%) of the 227 industry-sponsored RCTs reported the role of the sponsor. Of these, 20 explicitly stated that the sponsor had no role in the study. Twenty-one papers described the sponsor's role, the degree of sponsor involvement was variable and usually described vaguely. Among these papers, four stated that researchers had full access to all data, one that the researchers had no limits on publication and one that 'the decision to submit the paper for publication was determined by the study sponsor'. In conclusion, no researcher should be expected to produce 'findings' without full access to the data, freedom from interference in analysis and interpretation and liberty to publish all results, however disappointing to the stakeholder they may be. In the meantime, researchers do well to arm themselves with the rules for research partnerships and editors to take on the role of watchdog. PMID- 16214045 TI - Randomised study of tegafur and oral leucovorin versus intravenous 5-fluorouracil and leucovorin in patients with advanced colorectal cancer. AB - This randomised, open-label trial compared oral tegafur (FT)/leucovorin (LV) with the intravenous bolus 5-fluorouracil (5-FU)/LV as first-line chemotherapy for advanced colorectal cancer (CRC). Patients were randomised to receive oral FT 750 mg/m2/day for 21 days and LV 15 mg/m2 every 8 h in cycles repeated every 28 days (n=114), or intravenous LV 20 mg/m2 followed by 5-FU 425 mg/m2 daily for 5 days every 4 weeks for 2 cycles, and later every 5 weeks (n=123). Response rate was significantly higher in the FT/LV arm (27%, 95% CI 19-35) than in the 5-FU/LV arm (13%, 95% CI 7-19) (p<0.004). The median time to progression was 5.9 months (95% CI, 5.3-6.5; FT/LV arm) and 6.2 months (95% CI, 5.4-6.9; 5-FU/LV arm). Median overall survival was 12.4 months (95% CI, 10.3-14.5 months; FT/LV arm) and 12.2 months (95% CI, 8.9-15.7 months; 5-FU/LV arm) (p=n.s.; hazard ratio FT/LV:5 FU/LV=1.02). 5-FU/LV showed a higher incidence of grade 3/4 neutropenia (4.1 vs. 0%). Non-hematological toxicities showed similar incidences in the two treatment arms. Oral FT/LV was more active than IV 5-FU/LV in terms of objective response rate with similar overall survival, and with a favorable toxicity profile. This makes FT/LV a valid alternative to the IV 5-FU schedule in CRC patients. PMID- 16214049 TI - Functional enhancement of skeletal muscle by gene transfer. PMID- 16214048 TI - Understanding skeletal muscle adaptation to exercise training in humans: contributions from microarray studies. PMID- 16214051 TI - Amyotrophic lateral sclerosis microgenomics. PMID- 16214050 TI - Cell therapy for muscle regeneration and repair. PMID- 16214053 TI - Single muscle fiber physiology in neuromuscular disease. PMID- 16214052 TI - Redox mechanisms of muscle dysfunction in inflammatory disease. PMID- 16214054 TI - Electrodiagnostic studies in a murine model of demyelinating Charcot-Marie-Tooth disease. PMID- 16214055 TI - Using electromyography to assess function in humans and animal models of muscular dystrophy. PMID- 16214056 TI - Neurotrophic factors in neuromuscular disease. PMID- 16214057 TI - Electrodiagnostic automation: principles and practice. PMID- 16214058 TI - Physiological and anatomical basis of muscle magnetic resonance imaging. PMID- 16214060 TI - Approaching fatigue in neuromuscular diseases. PMID- 16214059 TI - Obesity, physical activity, and the metabolic syndrome in adult neuromuscular disease. PMID- 16214061 TI - The role of microglial cells in amyotrophic lateral sclerosis. PMID- 16214062 TI - Skeletal muscle in amyotrophic lateral sclerosis: emerging concepts and therapeutic implications. PMID- 16214063 TI - Chronic pain in persons with neuromuscular disease. PMID- 16214064 TI - Exploring chronic pain in youths with Duchenne Muscular Dystrophy: a model for pediatric neuromuscular disease. PMID- 16214065 TI - Respiratory support of individuals with Duchenne muscular dystrophy: toward a standard of care. PMID- 16214066 TI - Antidepressants in rheumatic disorders: do they act as analgesics or antidepressants? PMID- 16214067 TI - Osteoporosis in patients with inflammatory bowel disease. PMID- 16214068 TI - Interleukin-1: a new therapeutic target for ankylosing spondylitis? PMID- 16214069 TI - Hyperalgesia and allodynia: peripheral mechanisms. AB - Nociceptive signals are generated by peripheral sensory organs called nociceptors, which are endings of small-diameter nerve fibers responsive to the tissue environment. The myriad chemical mediators capable of activating, sensitizing, or arousing nociceptors include kinins, proinflammatory and anti inflammatory cytokines, prostanoids, lipooxygenases, the "central immune response mediator" NF-kappaB, neurotrophins and other growth factors, neuropeptides, nitric oxide, histamine, serotonin, proteases, excitatory amino acids, adrenergic amines, and opioids. These mediators may act in combination or at a given time in the inflammatory process, producing subtle changes that result in hyperalgesia or allodynia. We will review the most extensively studied molecular and cellular mechanisms underlying these two clinical abnormalities. The role of the peripheral nervous system in progression of inflammatory joint disease to chronicity is discussed. PMID- 16214070 TI - Osteoporotic fractures of the proximal humerus, pelvis, and ankle: epidemiology and diagnosis. AB - Although fractures involving the wrist, spine, and proximal femur are known to be strongly associated with osteoporosis, the underlying bone insufficiency often receives insufficient diagnostic and therapeutic attention. Osteoporosis also increases the risk of fractures at other sites. Low-energy fractures in patients older than 50 years should lead to investigations for osteoporosis, the only exceptions being fractures of the skull, cervical spine, fingers, and toes. The incidence rates of fractures of the proximal humerus, pelvis, and ankle are climbing relentlessly. Whereas fractures of the proximal humerus and pelvis are undoubtedly related to osteoporosis, the link is less well established for fractures of the ankle. Mortality and morbidity rates associated with pelvic fractures are similar to those seen with fractures of the proximal femur, in keeping with the fact that both fractures occur in elderly individuals. PMID- 16214071 TI - Phosphate diabetes, tubular phosphate reabsorption and phosphatonins. AB - Phosphate diabetes is defined as inadequate tubular reabsorption. Hypophosphatemia is responsible for most of the clinical manifestations, which vary with the age of the patient and the severity of the phosphate wasting. Vitamin D-resistant rickets in children or osteomalacia in adults, osteoporosis, bone pain including spinal pain, and pain in the joints and periarticular areas are the main manifestations. Several factors are known to affect tubular phosphate reabsorption via the sodium/phosphate cotransporters located on the tubular cell membranes. Factors that decrease phosphate reabsorption include a high intake of dietary phosphate, acidosis, parathyroid hormone (PTH), PTH related peptide (PTHrp), glucocorticoid therapy, calcitonin, and vitamin D. On the other hand, a low-phosphate diet, alkalosis, growth hormone, insulin, IGF-1, and thyroid hormones increase tubular phosphate reabsorption. Physiological concepts about tubular phosphate reabsorption have been radically changed by the recent identification of phosphaturic factors called phosphatonins. The most extensively studied phosphatonin to date is fibroblast growth factor 23 (FGF23), which was first identified in patients with tumor-induced osteomalacia and shown to be secreted by the neoplastic cells. The FGF23 has also been implicated in autosomal dominant hypophosphatemic rickets, in which a gene mutation results in production of abnormal FGF23 that resists hydrolysis. In healthy individuals, FGF23 contributes to regulate phosphate reabsorption via Na/Pi cotransporters. Other phosphatonins may exist, such as matrix extracellular phosphoglycoprotein (MEPE) and secreted frizzled-related protein 4 (SFRP4), whose role remains to be defined. The part played by these proteins in idiopathic renal phosphate wasting in adults needs to be investigated. PMID- 16214072 TI - Pathophysiology and clinical presentations of rhabdomyolysis. AB - Rhabdomyolysis has sparked new interest in recent years. The causes of rhabdomyolysis include drugs and other toxic agents, infections, physical exertion, crush injury, and muscle diseases (dystrophinopathies and metabolic myopathies). Prompt identification of the pathophysiological mechanism is the key to rapid control of the acute episode and to prevention of recurrences. In this update, we discuss the pathophysiological mechanisms and nosology of rhabdomyolysis, as well as diagnostic investigations, with special emphasis on noninvasive methods. PMID- 16214073 TI - Evaluation of depression and anxiety in patients with systemic sclerosis. AB - OBJECTIVE: To determine the prevalence of depression and anxiety in patients receiving follow-up in France for systemic sclerosis. PATIENTS: We prospectively evaluated 42 patients admitted for a follow-up evaluation of systemic sclerosis, including 18 with diffuse cutaneous scleroderma and 24 with limited cutaneous scleroderma. Patients admitted for recent organ involvement were excluded. Mean age was 57 +/- 13 years, mean disease duration was 10.2 +/- 8 years, and the mean functional Health Assessment Questionnaire score was 0.682 +/- 0.649. Only four patients had a history of antidepressive drug therapy. We used the Montgomery Asberg Depression Rating Scale (MADRS) and the Hamilton Anxiety Rating Scale to evaluate depression and anxiety, respectively. RESULTS: Eighteen (43%) patients met criteria for depression and 11 (26%) had scores above the cutoff usually taken to define moderate-to-severe depression. Twenty-seven (64%) patients met criteria for minor anxiety and eight (19%) for major anxiety. Depression and anxiety were strongly correlated with each other (r = 0.89; P < 0.0001). The MADRS score was significantly higher in the patients with pulmonary restrictive disease (P = 0.009) but was not associated with the extent of skin involvement, organ involvement, or disability. CONCLUSION: Systemic scleroderma is associated with a high prevalence of depression and anxiety. These disorders should be looked for routinely and the need for specific treatment evaluated. PMID- 16214074 TI - Exploration of small fibers for testing diabetic neuropathies. AB - INTRODUCTION: Electrophysiological exploration of neuropathies is a standard method of investigating the dysfunction of myelinated larger fibers (Aalpha, Abeta). However, this method cannot test dysfunctions in other fibers. To evaluate the smaller (Adelta) and unmyelinated fiber (C-fibers) lesions a quantitative method has been perfected: the study of the sensory thresholds (quantitative sensory testing: QST). It allows the investigation of the sensory symptoms and is a reproducible, non-invasive and painless method. It is used above all in patients suffering from diabetic neuropathy ('Diabetes Care 9 (1987) 432'). PATIENTS AND METHODS: We used the QST testing in comparison with nerve conduction velocities in 40 Non-Insulin-Dependent Diabetes Mellitus (NIDDM or Type II) patients in their 60s (+/-10 years). Depending on the duration of their diabetes (dd), we distinguished three groups: dd < 5 years (GI) dd from 5 to 15 years (GII) and dd > 15 years (GIII). All the patients underwent a clinical neurological examination, which enabled us to establish a gravity score comparable to the NDS (Neuropathy Disability Score: 'Muscle Nerve 10 (1988) 21'). RESULTS: Nerve conduction velocities and QST were studied for each group of patients. Electrophysiological alterations were connected to the gravity clinical score and in some asymptomatic patients a higher QST heat threshold could be observed. DISCUSSION: These results indicate that QST can detect the early dysfunction of the unmyelinated fibers in this kind of neuropathy. Subclinical detection can reduce severe neurological complications and make possible an early and effective treatment. PMID- 16214075 TI - Effect of alendronate therapy in children with osteogenesis imperfecta. AB - OBJECTIVE: To evaluate the effect of orally administered alendronate in children with osteogenesis imperfecta. METHODS: Thirty children (16 girls and 14 boys; mean age at baseline 10.7 +/- 6.0 years; range 4-16 years) with osteogenesis imperfecta type I (n = 22), III (n = 2), or IV (n = 6) were treated with alendronate (5 mg/day in patients aged 4-10 years and 10 mg/day in children >10 years of age) for 3 years. RESULTS: After 1 year of alendronate therapy we observed a significant increase in areal and volumetric bone mineral density Z scores (from -2.03 +/- 1.51 to -1.04 +/- 1.20, and from -1.91 +/- 1.38 to -1.33 +/- 1.30, respectively, P < 0.001), together with a significant drop in fracture rate (from 3.77 +/- 1.57 to 0.13 +/- 0.57, P < 0.000001), relief of chronic pain (from 3.83 +/- 1.44 days of pain/week to 0.73 +/- 0.77, P < 0.000001) and improvement in ambulation/mobility (P < 0.00002). After additional 2 years of therapy there were no further significant changes in these parameters, however the improvement was still remarkable in comparison to the pretreatment values (P < 0.003, P < 0.004, P < 0.000001, P < 0.000001 and P < 0.00001, respectively). A significant drop in markers of bone turnover (urinary deoxypyridinoline and serum osteocalcin) occurred after 3 years of therapy (P < 0.003 and 0.004, respectively). No adverse reactions were observed throughout the treatment. CONCLUSIONS: Alendronate has positively influenced quality of life in paediatric patients with osteogenesis imperfecta. Bisphosphonate therapy should be used only in the context of a well-defined protocol. PMID- 16214076 TI - Tuberculosis of the greater trochanter: a report of three cases. AB - Trochanteric tuberculosis represents less than 2% of all musculo-skeletal tuberculosis. The diagnosis is difficult especially if abscess and fistula are missing. The authors report 3 cases of trochanteric tuberculosis. The diagnosis was established, respectively, 4, 9 months and 1 year after the beginning of the symptoms. The tuberculosis was plurifocal in all cases. Diagnosis was based on the presence of caseum granuloma in the first case, positive Lowenstein culture in the second case and on clinical and paraclinical arguments in the third one. Healing was obtained after medical treatment alone. The authors discuss the potential role of the newer imaging modalities in diagnosis of trochanteric tuberculosis and the indications of medical and surgical treatment. PMID- 16214077 TI - Anti-PCNA antibodies: prevalence and predictive value. PMID- 16214078 TI - Sciatica as the presenting symptom of pleural mesothelioma and non-Hodgkin lymphoma. PMID- 16214079 TI - Adoptive immunotherapy: new ways to skin the cat? PMID- 16214080 TI - C-reactive protein: ligands, receptors and role in inflammation. AB - C-reactive protein (CRP) is the prototypical acute phase serum protein, rising rapidly in response to inflammation. CRP binds to phosphocholine (PC) and related molecules on microorganisms and plays an important role in host defense. However, a more important role may be the binding of CRP to PC in damaged membranes. CRP increases clearance of apoptotic cells, binds to nuclear antigens and by masking autoantigens from the immune system or enhancing their clearance, CRP may prevent autoimmunity. CRP binds to both the stimulatory receptors, FcgammaRI and FcgammaRIIa, increasing phagocytosis and the release of inflammatory cytokines; and to the inhibitory receptor, FcgammaRIIb, blocking activating signals. We have shown that, in two animal models of systemic lupus erythematosus (SLE), the (NZB x NZW)F1 mouse and the MRL/lpr mouse, a single injection of CRP before onset of proteinuria delayed disease development and late treatment reversed proteinuria. Thus, in these models, CRP plays an anti-inflammatory role. PMID- 16214081 TI - Functional analysis of highly defined, FACS-isolated populations of human regulatory CD4+CD25+ T cells. PMID- 16214083 TI - Immunodysregulation of HIV disease at bone marrow level. AB - Hematological abnormalities frequently occur in patients infected with HIV-1. Increasing evidence indicates that bone marrow (BM) suppression results from viral infection of accessory cells, with impaired stromal function and alteration of hematopoietic growth factor network. We investigated the effects of antiretroviral therapy on cytokine and chemokine production by BM cells and stromal cells, in a group of HIV-1-infected subjects before and during treatment. Compared with uninfected controls, an altered cytokine and chemokine production by BM cells has been observed before treatment, characterised by decreased IL-2 and elevated TNF-alpha, MIP-1alpha, MIP-1beta, and RANTES levels, along with a defective BM clonogenic activity. Antiretroviral therapy determined an amelioration of stem cell activity, a restoration of stromal cell pattern and functions, and an increased IL-2 production at BM level and a decrease of Fas expression on progenitor cells, in parallel with the diminution of TNF-alpha levels. HIV-1 protease inhibitors (PIs) may improve hematopoietic functions owing to their direct effects on the BM progenitor cells. Ritonavir and indinavir increased the colony growth of BM obtained either from HIV-1-infected patients or from normal individuals, in parallel with the normalization of functional and morphologic characteristics of stromal cells. PMID- 16214084 TI - CD4+ regulatory T cells: mechanisms of induction and effector function. AB - The main subsets of CD4+ regulatory T (Tr) cells are the CD4+ CD25+ Tr cells and the type 1 regulatory (Tr1) cells. Both subsets are essential for the maintenance of peripheral tolerance. CD4+ CD25+ Tr cells control immune homeostasis (e.g., in autoimmunity) mainly by cell contact-dependent interactions. In contrast, Tr1 cells regulate immune responses in transplantation, allergy, and autoimmune diseases, via an IL-10- and TGF-beta-dependent mechanism. Identification of the mechanisms responsible for the induction of the different Tr subsets in vivo is a matter of intense investigation. Studies on dendritic cells (DC), performed in the last years by several groups, have significantly contributed to clarify this point. Indeed, it is now clear that the role of DCs is not only to sense danger, but also to tolerize the immune system to antigens (Ags) encountered in the absence of maturation/inflammatory stimuli. Therefore, if a naive T cell encounters the antigen on immature DCs (iDCs), it differentiates into a Tr cell rather than an effector T cell. A better understanding of the mechanisms underlying the induction and functions of Tr cells in controlling the immune system is critical in view of a future cellular therapy to modulate immune mediated pathologies. PMID- 16214085 TI - Altered circadian rhythms in rheumatoid arthritis patients play a role in the disease's symptoms. AB - The circadian changes in the metabolism or nocturnal secretion of endogenous corticosteroids (reduction) observed in rheumatoid arthritis (RA) patients are responsible, in part, for the time-dependent changes that are observed in the inflammatory response and related early morning clinical symptoms of the disease. Melatonin (MLT), another circadian nocturnal hormone that is the secretory product of the pineal gland, has been implicated in the time-dependent RA inflammatory reaction with effects that are opposite to those of corticosteroids. As a consequence, altered functioning of the HPA axis (early morning reduced corticosteroid production) and of the pineal gland (night increased MLT production) found in RA patients, seem to be important factors in the appearance and perpetuation of the clinical circadian symptoms of the disease. Consistently, human proinflammatory Th1-type cytokine production (related to MLT stimulation) exhibits a diurnal rhythmicity with peak levels during the night and early morning, at a time when plasma cortisol (inducing the Th2-type cytokine production) is lowest and MLT is highest. Reduced daily light exposure as observed in northern Europe (Estonia), at least during the winter, might explain the higher and more prolonged serum MLT concentrations that were observed in northern RA patients, as well as some epidemiological features versus southern Europe patients. PMID- 16214086 TI - Immuno-tolerogenic functions of HLA-G: relevance in transplantation and oncology. AB - Over the past few years, the non-classical HLA class I molecule HLA-G has been the center of intense investigations that have led to the description of its particular structural and functional properties. Although located in the HLA class I region of chromosome six, the HLA-G gene may be distinguished from other HLA class I genes by its low polymorphism and alternative splicing that generates seven HLA-G proteins, whose tissue-distribution is restricted to normal fetal and adult tissues that display a tolerogeneic function towards both innate and acquired immune cells. We review here the functions of HLA-G from an immunological stand point, focusing on the recent advances in this field and on their potential impact in human pathologies, such as cancer and organ transplantation. PMID- 16214087 TI - Single nucleotide polymorphisms detection based on DNA microarray technology: HLA as a model. AB - The significance of DNA variations among individuals, including single nucleotide polymorphisms (SNPs) and/or genome nucleotide mutations as well as to their detection by using new technology, will improve and facilitate the knowledge of each gene sequence. Microarray may provide information about thousands of gene simultaneously, leading to a more rapid and accurate genotyping. In this view, we developed a new methodology as an example for the detection of SNPs based on DNA microarray, using a panel of HLA alleles representative of loci A, B, DRB1. A panel of 180 oligonucleotide probes was selected to identify polymorphic positions located in exons 2 and 3 of HLA-A and B, and in exon 2 of HLA-DRB1 locus. Each oligonucleotide sequence was designed with a nucleotide mismatch located in the same position as the center of the hybridization sequence. Hybridization experiments were carried out with genomic probes constructed with an asymmetric PCR strategy. The amplified DNAs were obtained from bone marrow cells of donors previously typed for transplant. The results obtained were showing that the method was reliable thus providing a feasible technique both for HLA typing and for the investigation of other regions of genetic and clinical interest including polymorphisms correlated with different autoimmune diseases. PMID- 16214088 TI - Manipulation of immune regulation in systemic lupus erythematosus. AB - Production of autoantibodies by B cells in systemic lupus erythematosus (SLE) can be interrupted via induction of regulatory and suppressor T cells. We have used the strategy of tolerizing lupus-prone (NZBxNZW)F(1) mice with an artificial peptide based on sequences common to several anti-double stranded (ds)DNA antibodies to induce regulatory and suppressor T cells that block production of anti-DNA antibodies and prolong their survival. At least one type of suppressor T cells (CD8+) and one type of regulatory T cell (CD4+ expressing the IL-2 receptor alpha chain CD25) are raised under this condition. While CD8+ suppressors (TS) require soluble factors to block help of T cells to B cells, regulatory CD4+CD25+ T cells (TR) curb the production of anti-DNA antibodies from B cells via cell contact through molecules that include membrane-bound TGFbeta and GITR. Moreover, CD8+ suppressors seem to act independently on antigen specificity, while TR act in an antigen-specific fashion. We hypothesize that the differences between these two lymphocyte subsets that share the common ability to dampen production of autoantibodies might underlie significant temporal and teleological advantages for optimal control of autoimmune reactivity. PMID- 16214089 TI - NK-DC interaction: on the usefulness of auto-aggression. AB - In recent years a number of studies have highlighted the novel concept that the actual role of natural killer (NK) cells is not only confined to the destruction of virus-infected cells or tumors. Indeed NK cells, by interacting with myeloid DCs during the early phases of inflammation, appear to play a crucial role in shaping both innate immune reactions (within inflamed peripheral tissues) and adaptive immune responses (in secondary lymphoid compartments). Interestingly, this novel function assigned to NK cells is essentially mediated through the aggression of normal immature myeloid DCs. Only DCs undergoing optimal maturation become refractory to NK cell killing and will obtain the permission to prime Th1 cells after migration to lymph nodes. PMID- 16214090 TI - CD20: a target antigen for immunotherapy of autoimmune diseases. AB - This article reviews the role of CD20 antigen in B cell function and the effectiveness and limits of passive immunotherapy with anti-CD20 monoclonal antibody (Rituximab) in the treatment of autoimmune (or immune-mediated) diseases. Active immunotherapy is a more feasible way to control these chronic diseases. A peptide that mimics the CD20 epitope recognized by Rituximab is employed to stimulate the host immune response against CD20. PMID- 16214091 TI - Cyclosporin A regulates human NK cell apoptosis induced by soluble HLA-I or by target cells. AB - Human natural killer (NK) cells are effectors of innate immunity, capable of killing transformed or virus-infected cells and producing pro-inflammatory cytokines. Soluble molecules of HLA-I (sHLA-I), which are significantly increased in the serum of patients affected by auto-immune or infectious or neoplastic diseases, induce NK cell apoptosis interacting with its ligands, such as CD8 or the activating isoforms of members of inhibitory superfamily receptors (IRS). This cell death is accompanied by the release of large amounts of interferon gamma. NK cells can kill autologous target cells, including antigen presenting cells or infected or tumor cells, by engaging the natural cytotoxicity receptors (NCR) NKp30, or NKp44 and NKp46. Again, the binding between NCR on NK cells and their putative ligands on targets leads to NK cell apoptosis. FasL produced and secreted by NK cells is responsible for the NK cell apoptosis induced by either HLA-I receptors or NCR. Interestingly, cyclosporin A (CsA) blocks NK cell death consequent to interaction with target cells or with sHLA-I, without affecting the activation of cytolysis. This would indicate that CsA can maintain NK cell dependent innate immunity by prolonging NK cell survival in an hostile environment in the presence of sHLA-I or target cells. PMID- 16214092 TI - Emerging biologic drugs for the treatment of rheumatoid arthritis. AB - This article reviews the role of emerging biologic drugs for the treatment of rheumatoid arthritis (RA). Besides anti-tumor necrosis factor (TNF)-alpha and anti-interleukin (IL)-1 agents (Infliximab, Adalimumab, Etanercept and Anakinra) whose clinical efficacy is now established, new drugs have been proposed for the therapy of rheumatoid arthritis patients not responding to conventional treatments. These approaches include the blockade of B-cell activity with anti CD20 monoclonal antibody (Rituximab) and the inhibition of T-cell activation with fusion protein CTLA4Ig. Moreover, promising results have been obtained in animal models utilizing suppressors of cytokine signaling (SOCS) and dominant-negative TNF variants to inactivate TNF signaling. PMID- 16214093 TI - Th1 cytokines in the pathogenesis of lupus nephritis: the role of IL-18. AB - Excessive T helper cell function is a hallmark of systemic lupus erythematosus and abnormalities of T helper cytokine profiles have been implicated in loss of immune tolerance, increased antigenic load, defective B cell suppression and a variety of clinical manifestations. Here, we emphasize the importance of T helper 1-type (i.e., IFN-gamma) with respect to T helper 2-type (i.e., IL-4) cytokines in promoting the pathogenesis of lupus nephritis focusing on the critical role of IL-18, a major T helper 1 differentiation factor. IL-18 is overexpressed in patients with lupus nephritis along with higher INF-gammaand lower IL-4 production as compared to non-nephritic lupus patients. We hypothesize a pathogenic model where both the onset and aggravation of nephritis are promoted by an imbalance of immune response towards a T helper 1 cytokine predominance due to IL-18 up-regulation. In contrast, a T helper 2-skewed cytokine profile may possibly prevent the development of renal disease. In this context, IL-18 represents a novel marker of lupus nephritis and its measurement may be helpful in the assessment of patients. PMID- 16214094 TI - B-cell differentiation in the CNS of patients with multiple sclerosis. AB - Clonally expanded populations of Ig variable gene-mutated B cells are found in the central nervous system (CNS) of subjects with multiple sclerosis (MS), suggesting the occurrence of a germinal center-like reaction. Recent studies have demonstrated that the cerebrospinal fluid (CSF) of MS patients is enriched with centroblasts and B cells with a memory phenotype compared to peripheral blood. In the same individuals, antibody-secreting cells (ASC) are detected in the CSF and appear to correlate with CNS inflammation. These B-cell subsets are the output of a germinal center reaction, which is likely to occur in the CNS. Recent findings suggest that the inflamed brain can become a favorable niche for B-cell survival and proliferation and, under some circumstances, sustain the formation of ectopic lymphoid structures. Thus, B cells are likely to expand and mature inside the CNS, giving rise to ASC, which may play an effector role in the pathogenesis of MS. PMID- 16214095 TI - Environmental adjuvants, apoptosis and the censorship over autoimmunity. AB - Alterations during apoptosis lead to the activation of autoreactive T cells and the production of autoantibodies. This article discusses the pathogenic potential of cells dying in vivo, dissecting the role of signals that favor immune responses (adjuvants) and the influence of genetic backgrounds. Diverse factors determine whether apoptosis leads or not to a self-sustaining, clinically apparent autoimmune disease. The in vivo accumulation of uncleared dying cells per se is not sufficient to cause disease. However, dying cells are antigenic and their complementation with immune adjuvants causes lethal diseases in predisposed lupus-prone animals. At least some adjuvant signals directly target the function and the activation state of antigen presenting cells. Several laboratories are aggressively pursuing the molecular identification of endogenous adjuvants. Sodium monourate and the high mobility group B1 protein (HMGB1) are, among those identified so far, well known to rheumatologists. However, even the complementation of apoptotic cells with potent adjuvant signals fail to cause clinical autoimmunity in most strains: autoantibodies generated are transient, do not undergo to epitope/spreading and do not cause disease. Novel tools for drug development will derive from the molecular identification of the constraints that prevent autoimmunity in normal subjects. PMID- 16214096 TI - The immune response to citrullinated antigens in autoimmune diseases. AB - Post-translational modifications of proteins occur very frequently. One of these modifications, citrullination, is the result of arginine deimination operated by an enzyme, peptidylarginine deiminase (PAD), whose activity is under strict genetic control. Serum antibodies reactive with citrullinated proteins/peptides are a very sensitive and specific marker for rheumatoid arthritis. Genes encoding for PAD enzymes have been investigated in RA: the PADI4 gene confers susceptibility to RA in Japanese patients, but not in Caucasians. PMID- 16214097 TI - Antigen-specific down-regulation of immunopathological events in an experimental autoimmune kidney disease. AB - Heymann nephritis (HN) is an experimental autoimmune disease of rats characterized by immune-complex (IC) depositions on the epithelial side of the glomerular basement membrane (GBM) and by proteinuria. Several forms of HN have been produced by various investigators, but one thing has been common to all of them, namely their inducement by the development of pathogenic IgG autoantibodies (aabs). The aim of this review is to describe how pathogenic IgG aab production (which initiates and maintains the disease) in slowly progressive HN (SPHN) can be specifically terminated by injections of ICs made up of native rat renal tubular antigens and IgM antibodies directed against them. PMID- 16214098 TI - T for two: when helpers need help. AB - This article discusses forms of cell cooperation involving CD4 T cells and their impact on adaptive immunity. In particular, evidence will be presented in favor of cooperation among CD4 T cells of different specificity, its molecular basis and functional significance. Th-Th cooperation, or help for helpers, will be discussed in relation to the induction of anti-tumor responses and, in converse, the generation of autoimmunity. A model is proposed where Th-Th cooperation may represent the structure/function substrate for the control of self/non-self discrimination and peripheral tolerance in vivo. PMID- 16214099 TI - Thymic regulatory T cells. AB - Several types of T regulatory (Treg) cells have been described in both mice and humans, including natural or professional (CD4+CD25+ T cells) and adaptive (Th3 and Tr1 cells) Treg cells. The former develops in the thymus and results in an endogeneous long-lived population of self-antigen-specific T cells in the periphery poised to prevent potentially autoimmune reactions. The second subset develops as a consequence of activation of mature T cells under particular conditions of sub-optimal antigen exposure and/or costimulation. Natural Treg cells are positively selected in the cortex through their TCR interactions with self-peptides presented by thymic stromal cells. It is likely that this high affinity recognition results in signals rendering them anergic and able to produce anti-apoptoptic molecules which protect them from negative selection. Recently, small subsets of CD4+CD25+ and of CD8+CD25+ cells sharing similar characteristics have been detected in human fetal and post-natal thymuses. Both CD4+CD25+ and CD8+CD25+ human thymocytes express Foxp3 and GITR mRNA, as well as surface CCR8 and TNFR2 and cytoplasmic CTLA-4 proteins, which are common features of mature Treg cells. Following activation they do not proliferate or produce cytokines, but express surface CTLA-4 and TGF-beta1. They suppress the proliferation of autologous CD4+CD25- thymocytes to allogeneic stimulation by a contact-dependent mechanism related to the combined action of surface CTLA-4 and TGF-beta leading to the inhibition of the IL-2R alpha chain on target T cells. Lastly, both CD4+CD25+ and CD8+CD25+ Treg thymocytes exert strong suppressive activity on Th1, but much lower on Th2 cells, since these latter may escape from suppression via their ability to respond to growth factors other than IL-2. Treg cells that develop in, and emerge from, the thymus are certainly responsible for the maintenance of self-tolerance and prevention of autoimmune disorders. The result that Th1 cells are highly susceptible to the suppressive activity of Treg thymocytes is consistent with the important role of these cells in protecting against the Th1-mediated immune response to autoantigens. PMID- 16214100 TI - Results of silicone oil removal in patients with cytomegalovirus retinitis related retinal detachments. AB - PURPOSE: To evaluate the percentage and risk indicators leading to retinal redetachment in HIV (human immunodeficiency virus) patients with CMV (cytomegalovirus) retinitis related retinal detachments that were repaired with silicone oil, and then subsequently underwent oil removal. DESIGN: Retrospective, noncomparative interventional case series. METHODS: The study cohort consisted of a series of 15 eyes in 14 patients with HIV and CMV retinitis with a retinal detachment (RD) repaired with silicone oil at a single center and followed from the time of the CMV retinitis diagnosis through the time of silicone oil removal. Patient- and eye-specific data regarding demographic and clinical characteristics were collected retrospectively and statistical analyses were performed to compare differences between the eyes that had retinal detachments versus the eyes that remained attached following removal of silicone oil. RESULTS: Eight eyes (53%) redetached after a median of 4.0 months following oil removal. Cataract surgery performed at the time of oil removal was a statistically significant risk factor for redetachment (P = .01). There was a trend for lower CD4 levels to be associated with a higher risk of retinal redetachment. The use of a scleral buckle at the time of surgery (initial RD repair or at the time of oil removal) did not reduce the risk of redetachment. CONCLUSIONS: Approximately half of the eyes with CMV related retinal detachment may safely undergo oil removal. The risk factor for redetachment was simultaneous cataract extraction at the time of silicone oil removal. There was also a trend for lower CD4 levels to be associated with a higher risk of retinal redetachment. PMID- 16214102 TI - Frontalis suspension for upper eyelid ptosis: evaluation of different surgical designs and suture material. AB - PURPOSE: To compare two sling designs (single loop or double pentagon) and a variety of suture material that was used in frontalis suspension surgery for correction of upper eyelid ptosis. DESIGN: Retrospective, nonrandomized, comparative interventional case series. METHODS: Medical record review of 99 patients (164 surgeries) who underwent frontalis suspension surgery for upper eyelid ptosis was conducted at the Jules Stein Eye Institute in 1996 to 2002. Functional and cosmetic success, margin reflex distance (MRD) and lagophthalmos were evaluated. RESULTS: MRD increased an average of 1.1 mm after the operation (P < .001). Ptosis recurrence was noticed in 42 cases (26%); polytetrafluoroethylene achieved the lowest recurrence rate (15%), although not statistically significant. No difference in functional success, ptosis recurrence, or change in MRD was noticed between single loop and double pentagon design. A better cosmetic outcome was noted in cases in which nylon suture was used. Complications included four cases (2.4%) of over-correction, three cases (1.8%) of suture infection (all in polytetrafluoroethylene), two cases of pyogenic granuloma (1.2%), and two cases (1.2%) of suture exposure. CONCLUSION: Frontalis suspension for upper eyelid ptosis resulted in 26% ptosis recurrence after a mean of 12 months from first surgery. Polytetrafluoroethylene showed the lowest incidence of ptosis recurrence. No statistically significant difference was found between different suture materials or loop shape that was used in the surgical technique. A better cosmetic outcome, as graded by different observers, was noted in cases in which a nylon sling was used. PMID- 16214101 TI - Phenotypic characterization of a large family with RP10 autosomal-dominant retinitis pigmentosa: an Asp226Asn mutation in the IMPDH1 gene. AB - PURPOSE: To evaluate the clinical features associated with the RP10 form of autosomal-dominant retinitis pigmentosa in 11 affected members of various ages from one family with a defined IMPDH1 mutation (Asp226Asn). DESIGN: Prospective, observational case series. METHODS: Visual function assessment included visual acuity, color vision, visual field, dark adaptometry, full-field electroretinography (ffERG), and multifocal electroretinography (mfERG). Ophthalmologic examinations, fundus photography, and optical coherence tomographic scans were also performed. Blood samples were obtained to screen for basic immune function. RESULTS: Visual acuity was slightly reduced in the teenage years and substantially reduced in association with cystoid macular edema (CME) at all ages. Color defects were observed in three patients (one teen, two adults). Dark-adapted thresholds were elevated. Visual fields were markedly constricted by age 40 (mPrPp>bPrPp>pAntp. The membrane perturbation effects of the N-terminal prion peptides suggest that they form transient pores (similar to melittin) causing toxicity in parallel with their cellular trafficking. PMID- 16214106 TI - Porous membranes for reconstitution of ion channels. AB - Functional biological synthetic composite (BSC) membranes were made using phospholipids, biological membrane proteins and permeable synthetic supports or membranes. Lipid bilayers were formed on porous polycarbonate (PC), polyethylene terephthalate (PETE) and poly (l-lactic acid) (PLLA) membranes and in 10-100 microm laser-drilled pores in a 96-well plastic plate as measured by increased resistance or decreased currents. Bilayers in 50 microm and smaller pores were stable for up to 4 h as measured by resistance changes or a current after gramicidin D reconstitution. Biological membrane transport reconstitution was then carried out. Using vesicles containing Kv1.5 K(+) channels, K(+) currents and decreased resistance were measured across bilayers in 50 microm pores in the plastic plate and PLLA membranes, respectively, which were inhibited by compound B, a Kv1.5 K(+) channel inhibitor. Functional reconstitution of Kv1.5 K(+) channels was successful. Incorporation of membrane proteins in functional form in stable permeable membrane-supported lipid bilayers is a simple technology to create BSC membranes that mimic biological function which is readily adaptable for high throughput screening. PMID- 16214107 TI - Peroxynitrite efficiently mediates the interconversion of redox intermediates of myeloperoxidase. AB - Nitric oxide-derived oxidants (e.g., peroxynitrite) are believed to participate in antimicrobial activities as part of normal host defenses but also in oxidative tissue injury in inflammatory disorders. A similar role is ascribed to the heme enzyme myeloperoxidase (MPO), the most abundant protein of polymorphonuclear leukocytes, which are the terminal phagocytosing effector cells of the innate immune system. Concomitant production of peroxynitrite and release of millimolar MPO are characteristic events during phagocytosis. In order to understand the mode of interaction between MPO and peroxynitrite, we have performed a comprehensive stopped-flow investigation of the reaction between all physiological relevant redox intermediates of MPO and peroxynitrite. Both iron(III) MPO and iron(II) MPO are rapidly converted to compound II by peroxynitrite in monophasic reactions with calculated rate constants of (6.8+/ 0.1) x 10(6) M(-1)s(-1) and (1.3+/-0.2) x 10(6) M(-1)s(-1), respectively (pH 7.0 and 25 degrees C). Besides these one- and two-electron reduction reactions of peroxynitrite, which produce nitrogen dioxide and nitrite, a one-electron oxidation to the oxoperoxonitrogen radical must occur in the fast monophasic transition of compound I to compound II mediated by peroxynitrite at pH 7.0 [(7.6+/-0.1) x 10(6) M(-1)s(-1)]. In addition, peroxynitrite induced a steady state transition from compound III to compound II with a rate of (1.0+/-0.3) x 10(4) M(-1)s(-1). Thus, the interconversion among the various oxidation states of MPO that is prompted by peroxynitrite is remarkable. Reaction mechanisms are proposed and the physiological relevance is discussed. PMID- 16214108 TI - Targeted inhibition of p38alpha MAPK suppresses tumor-associated endothelial cell migration in response to hypericin-based photodynamic therapy. AB - Photodynamic therapy (PDT) is an established anticancer modality and hypericin is a promising photosensitizer for the treatment of bladder tumors. We show that exposure of bladder cancer cells to hypericin PDT leads to a rapid rise in the cytosolic calcium concentration which is followed by the generation of arachidonic acid by phospholipase A2 (PLA2). PLA2 inhibition significantly protects cells from the PDT-induced intrinsic apoptosis and attenuates the activation of p38 MAPK, a survival signal mediating the up-regulation of cyclooxygenase-2 that converts arachidonic acid into prostanoids. Importantly, inhibition of p38alpha MAPK blocks the release of vascular endothelial growth factor and suppresses tumor-promoted endothelial cell migration, a key step in angiogenesis. Hence, targeted inhibition of p38alpha MAPK could be therapeutically beneficial to PDT, since it would prevent COX-2 expression, the inducible release of growth and angiogenic factors by the cancer cells, and cause an increase in the levels of free arachidonic acid, which promotes apoptosis. PMID- 16214109 TI - Binding of intimin with Tir on the bacterial surface is prerequisite for the barrier disruption induced by enteropathogenic Escherichia coli. AB - Enteropathogenic Escherichia coli (EPEC) infects intestinal epithelial cells and perturbs the intestinal barrier that limits the paracellular movement of molecules. The disruption of the barrier is mediated by the effectors translocated into the host cells through the bacterial type III secretion system (TTSS). A previous report has described the importance of a bacterial outer membrane protein, intimin, in EPEC-mediated disruption of the barrier, and proposed that intimin, in concert with a host intimin receptor, controls the activity of the translocated barrier-disrupting effectors [P. Dean, B. Kenny, Intestinal barrier dysfunction by enteropathogenic Escherichia coli is mediated by two effector molecules and a bacterial surface protein, Mol. Microbiol. 54 (2004) 665-675]. In this study, we found that the importance of intimin is in its ability to bind a bacterial intimin receptor, Tir. Additionally, the impaired ability of an intimin-negative mutant was not restored by co-infection with intimin-expressing TTSS mutants. Collectively, the results in this study favor an alternative scenario explaining the importance of intimin, that the binding of intimin with Tir on the bacterial surface triggers or promotes the translocation of factors required for the efficient disruption of the barrier. Thus, the interaction of intimin with Tir may serve as a molecular switch that controls the delivery of virulence factors into the host cells. PMID- 16214110 TI - Stalk region of kinesin-related protein Unc104 has moderate ability to form coiled-coil dimer. AB - Unc104/KIF1A, a kinesin family member, is reported to be monomeric in solution, though its polypeptide has regions that potentially form coiled coils. For a better understanding of the mechanism underlying Unc104/KIF1A's motility, it is important to evaluate the dimerization ability of this protein. The CD measurement of relevant segments of Caenorhabditis elegans Unc104 indicated that peptides having a common region (N358-K379) showed spectra characteristic to an alpha-helix. Dimerization by coiled-coil formation was confirmed by analytical ultracentrifugation. By analyzing the concentration dependence of the CD spectra, the monomer-dimer dissociation constant, Kd, of (N354-E388) was estimated to be about 5 microM, which is considerably larger than that of the corresponding segment of human kinesin (62 nM). Though its dimerization ability is rather moderate, Unc104/KIF1A could nonetheless dimerize and therefore could move by the same mechanism as human kinesin when the concentration of Unc104 is high due to, e.g., local crowding. This suggests that the motility could be controlled by the concentration of the motor protein. PMID- 16214111 TI - Identification and characterisation of the Plasmodium vivax rhoptry-associated protein 2. AB - Plasmodium vivax is currently the most widespread of the four parasite species causing malaria in humans around the world. It causes more than 75 million clinical episodes per year, mainly on the Asian and American continents. Identifying new antigens to be further tested as anti-P. vivax vaccine candidates has been greatly hampered by the difficulty of maintaining this parasite cultured in vitro. Taking into account that one of the most promising vaccine candidates against Plasmodium falciparum is the rhoptry-associated protein 2, we have identified the P. falciparum rhoptry-associated protein 2 homologue in P. vivax in the present study. This protein has 400 residues, having an N-terminal 21 amino-acid stretch compatible with a signal peptide and, as occurs with its falciparum homologue, it lacks repeat sequences. The protein is expressed in asexual stage P. vivax parasites and polyclonal sera raised against this protein recognised a 46 kDa band in parasite lysate in a Western blot assay. PMID- 16214112 TI - Accumulation of infectious mutants in stocks during the propagation of fiber modified recombinant adenoviruses. AB - In infected cells, replication errors during viral proliferation generate mutations in adenoviruses (Ads), and the mutant Ads proliferate and evolve in the intracellular environment. Genetically fiber-modified recombinant Ads (rAd variants) were generated, by modification of the fiber gene, for therapeutic applications in host cells that lack or express reduced levels of the Coxsackievirus and adenovirus receptor. To assess the genetic modifications of rAd variants that might induce the instability of Ad virions, we examined the frequencies of mutants that accumulated in propagated stocks. Seven of 41 lines of Ad variants generated mutants in the stocks and all mutants were infectious. Moreover, all the mutations occurred in the modified region that had been added at the 3' end of the fiber gene. Our results show that some genetic modifications at the carboxyl terminus of Ad fiber protein lead to the instability of Ad virions. PMID- 16214113 TI - Prion proteins from susceptible and resistant sheep exhibit some distinct cell biological features. AB - It is well established that natural polymorphisms in the coding sequence of the PrP protein can control the expression of prion disease. Studies with a cell model of sheep prion infection have shown that ovine PrP allele associated with resistance to sheep scrapie may confer resistance by impairing the multiplication of the infectious agent. To further explore the biochemical and cellular mechanisms underlying the genetic control of scrapie susceptibility, we established permissive cells expressing two different PrP variants. In this study, we show that PrP variants with opposite effects on prion multiplication exhibit distinct cell biological features. These findings indicate that cell biological properties of ovine PrP can vary with natural polymorphisms and raise the possibility that differential interactions of PrP variants with the cellular machinery may contribute to permissiveness or resistance to prion multiplication. PMID- 16214114 TI - Identification of neuropeptides from the decapod crustacean sinus glands using nanoscale liquid chromatography tandem mass spectrometry. AB - Neurosecretory systems are known to synthesize and secrete a diverse class of peptide hormones which regulate many physiological processes. The crustacean sinus gland (SG) is a well-defined neuroendocrine site that produces numerous hemolymph-borne agents including the most complex class of endocrine signaling molecules--neuropeptides. As an ongoing effort to define the peptidome of the crustacean SG, we determine the neuropeptide complements of the SG of the Jonah crab, Cancer borealis, and the Maine lobster, Homarus americanus, using nanoflow liquid chromatography electrospray ionization quadrupole time-of-flight (ESI QTOF) MS/MS. Numerous neuropeptides were identified, including orcokinins, orcomyotropin, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptides (CPRPs), red pigment concentrating hormone (RPCH), beta-pigment dispersing hormone (beta-PDH), proctolin and HL/IGSL/IYRamide. Among them, two novel orcokinins were de novo sequenced from the SG of H. americanus. Three CPRPs including a novel isoform were sequenced in H. americanus. Four new CPRPs were sequenced from the SG of C. borealis. Our results show that structural polymorphisms in CPRPs (and thus the CHH precursors) are common in Dendrobranchiata as well as in Pleocyemata. The evolutionary relationship between the CPRPs is also discussed. PMID- 16214115 TI - Modulation of MDR1 gene expression in multidrug resistant MCF7 cells by low concentrations of small interfering RNAs. AB - MDR1 overexpression is one form of the multidrug resistance (MDR) phenotype, which can be acquired by patients initially responsive to chemotherapy. Because of the high toxicity of the inhibitors of P-glycoprotein (P-gp), the protein encoded by MDR1, attention has been focused on selective modulation of the MDR1 gene. Small interfering RNAs (siRNAs) were shown to be powerful tools for such a purpose, even when used at low concentrations (< or =20 nM) in order to avoid sequence nonspecific effects. Two siRNAs used at 20 nM were shown to lead to efficient down-regulation of MDR1 at the protein level (only ca. 20% total P-gp expression remaining) in the doxorubicin selected MCF7-R human cell line. Cell surface expression of P-gp was inhibited, leading to reversal of the drug efflux phenotype (about 40% reversal with the most efficient siRNA) and enhancement of chemosensitivity (about 35%). At the mRNA level, the down-regulation of MDR1 obtained with the most efficient siRNA increased from about 50% (5 nM siRNA) to 60% (10 or 20 nM). The advantage of using a combination of siRNAs instead of a single one has been suggested. PMID- 16214116 TI - Inhibitory effects of dimethylacetyl-beta-cyclodextrin on lipopolysaccharide induced macrophage activation and endotoxin shock in mice. AB - The potential use of hydrophilic cyclodextrins (CyDs) as an inhibitor for lipopolysaccharide (LPS) was examined. Of the five CyDs used in this study, dimethylacetyl-beta-cyclodextrin (DMA7-beta-CyD) had greater inhibitory activity than other CyDs against the production of nitric oxide (NO) and various proinflammatory cytokines including tumor necrosis factor-alpha (TNF-alpha) in murine macrophages stimulated with two serotypes of LPS and lipid A. The inhibitory effect of DMA7-beta-CyD on NO production was also observed in macrophages stimulated with lipoteichoic acid (LTA), but not peptidoglycan (PGN), polyinosinic-polycytidylic acid (poly I:C) or CpG oligonucleotide (CpG-ODN). Several studies have suggested that the inhibitory effects of DMA7-beta-CyD could be ascribed to the interaction with LPS. Simultaneous administration of DMA7-beta CyD not only intraperitoneally but also intravenously and intraperitoneal injection of aqueous solution containing LPS and d-galactosamine in murine endotoxin shock model suppressed fatality. Also, DMA7-beta-CyD decreased blood level of TNF-alpha as well as serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) in mice. In conclusion, DMA7-beta-CyD may have promise as a new therapeutic agent for endotoxin shock induced by LPS. PMID- 16214117 TI - Identification of protein kinase C isoforms involved in cerebral hypoxic preconditioning of mice. AB - Recently, accumulated studies have suggested that protein kinases C (PKC) play a central role in the development of ischemic-hypoxic preconditioning (I/HPC) in the brain. However, which types of PKC isoforms might be responsible for neuroprotection is still not clear, especially when the systematic investigation of PKC isoform-specific changes in brain regions was rare in animals with ischemic-hypoxic preconditioning. By using Western blot, we have demonstrated that the levels of cPKC betaII and gamma membrane translocation were increased in the early phase of cerebral hypoxic preconditioning. In this study, we combined the Western blot and immunostaining methods to investigate the effects of repetitive hypoxic exposure (H1-H4, n = 6 for each group) on membrane translocation and protein expression of several types of PKC isoforms, both in the cortex and hippocampus of mice. We found that the increased membrane translocation of nPKCepsilon (P < 0.05, versus normoxic H0) but not its protein expression levels in both the cortex and hippocampus during development of cerebral HPC in mice. However, there were no significant changes in both membrane translocation and protein expression levels of nPKCdelta, theta, eta, mu, and aPKC iota/lambda, zeta in these brain areas after hypoxic preconditioning. Similarly, an extensive subcellular redistribution of cPKCbetaII, gamma, and nPKCepsilon was observed by immunostaining in the cortex after three series of hypoxic exposures (H3). These results indicate that activation of cPKCbetaII, gamma, and nPKCepsilon might be involved in the development of cerebral hypoxic preconditioning of mice. PMID- 16214118 TI - Ganglion cell densities in normal and dark-reared turtle retinas. AB - In dark-reared, neonatal turtle retinas, ganglion cell receptive fields and dendritic trees grow faster than normal. As a result, their areas may become, on average, up to twice as large as in control retinas. This raises the question of whether the coverage factor of dark-reared ganglion cells is larger than normal. Alternatively, dark rearing may lead to smaller-than-normal cell densities by accelerating apoptosis. To test these alternatives, we investigated the effect of light deprivation on densities and soma sizes of turtle retinal ganglion cells. For this purpose, we marked these cells using retrograde labeling of fixed turtle retinas with DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate). Control turtles were maintained in a regular 12-h light/dark cycle from hatching until 4 weeks of age, whereas dark-reared turtles were maintained in total darkness for the same period. Ganglion cells in the control and dark reared retinas were found to be similar in density and soma sizes. These results show that the mean coverage factor of turtle dark-reared ganglion cells is larger than normal. PMID- 16214119 TI - Functional role of alpha1-adrenoceptors in the locus coeruleus: a microdialysis study. AB - The present study elucidates the role of alpha(1)-adrenoreceptors in the locus coeruleus (LC) using a dual-probe microdialysis in conscious rats. One probe sampled noradrenaline in the LC, whereas the second probe sampled noradrenaline in a main projection area, the prefrontal cortex (PFC). To investigate a possible tonic activation of LC neurons by alpha(1)-adrenoceptor, the alpha(1)-antagonist prazosin (10 microM) was infused into the LC. Extracellular noradrenaline in the LC decreased to about 50% of basal levels but no change of noradrenaline release was detected in the ipsilateral PFC. Next, the interaction between alpha(1)- and alpha(2)-adrenoceptors was investigated. Local administration of the alpha(2) adrenoceptor antagonist idazoxan (100 microM) into the LC increased the noradrenaline release in the LC to about 400%, whereas noradrenaline release in the PFC rose to 150% of basal levels. A similar effect was seen when the specific alpha(2A)-adrenoceptor antagonist BRL 44408 (10 microM) was infused: extracellular noradrenaline in the LC and PFC increased to about 400 and 120% of the basal levels, respectively. When infusions of idazoxan (100 microM) or BRL 44408 (10 microM) into the LC were combined with prazosin (10 microM), the excitatory effects of the alpha(2)-adrenoceptor antagonists on the release of noradrenaline were strongly suppressed in the LC as well as in the ipsilateral PFC. It is concluded that alpha(1)-adrenoreceptors are involved in the regulation of LC activity. Apparently, alpha(1)- and alpha(2)-adrenoceptors have opposite roles in their function as autoreceptors on LC cells. PMID- 16214120 TI - Cloning, expression, and characterization of an oligoxyloglucan reducing end specific xyloglucanobiohydrolase from Aspergillus nidulans. AB - An oligoxyloglucan reducing end-specific xyloglucanobiohydrolase from the filamentous fungus Aspergillus nidulans was cloned and expressed in Pichia pastoris as a secreted histidine-tagged protein and purified by affinity chromatography. The enzyme acts on xyloglucan oligomers and releases the first two glycosyl residue segments from the reducing end, provided that neither the first glucose nor the xylose attached to the third glucose residue from the reducing end is not further substituted. The enzyme has a specific activity of 7 U/mg at the pH optimum of 3 and at the temperature optimum of 42 degrees C. PMID- 16214121 TI - The effect of dimethyl sulfoxide on the function of cytochrome P450 2D6 in HepG2 cells upon the co-expression with NADPH-cytochrome P450 reductase. AB - HepG2 cells, a human hepatoma cell line, stably expressing NADPH-cytochrome P450 reductase (OR) and/or cytochrome P450 2D6 wild-type (CYP2D6-WT) or its variants (Pro34Ser, Gly42Arg, Arg296Cys and Ser486Thr) were established in the present study. The cultivation of HepG2 cells expressing CYP2D6-WT in the culture medium containing dimethyl sulfoxide (DMSO, 0.1% of final concentration) markedly increased the bufuralol (BF) 1''-hydroxylase activity compared with that of control cells when cultivated without DMSO. A similar effect was also observed in HepG2 cells stably expressing CYP2D6 and OR. The addition of hemin in place of DMSO to the culture medium resulted in no increase in the enzyme activity. Western blot analysis revealed that the levels of CYP2D6 protein were similar between DMSO-treated and non-treated HepG2 cells regardless of OR expression. Spectrophotometric analysis of reduced carbon monoxide-difference spectra of HepG2 cells expressing CYP2D6-WT and/or OR demonstrated that the addition of DMSO increased the peak height of functional CYP2D6 at 450 nm. These results suggest that the increase in CYP2D6 activity is attributable to the radical-scavenging effect of DMSO. The HepG2 cell lines stably expressing OR and CYP2D6 or its variants in combination with DMSO treatment may be useful for screening the cytotoxicity of chemical compounds which undergo oxidation by CYP2D6. PMID- 16214122 TI - Risk stratification of chest pain patients by point-of-care cardiac troponin T and myoglobin measured in the emergency department. AB - A prospective multicenter study including 1410 chest pain patients with suspected acute coronary syndromes was carried out to examine the predictive value of biological cardiac markers for adverse events measured by a point-of-care system. Admission cardiac troponin T (cTnT) and myoglobin were measured in parallel on a point-of-care system in the emergency department and -- together with CK-MB mass - on lab analyzers. In a one-year follow-up, cardiac and non-cardiac death, acute myocardial infarction, unstable angina pectoris and need for revascularization were registered. Median time between onset of symptoms and admission was 285 min; 172 patients (12.2%) had no event during follow-up. If the cTnT, measured either by the point-of-care system or a conventional lab analyzer, was >0.05 microg/L, then the chance of a cardiac event during the follow-up period was doubled (18% vs. 9%). Serial cTnT measurement did not add any further value to the predictive power of the admission cTnT. Myoglobin and CK-MB mass identified increasing risk with increasing concentration quartiles; cardiac event rates were 2.8- to 4.4 fold higher between the quartiles with the lowest and those with the highest analyte concentration, respectively. There was no difference in non-cardiac death rates between any concentration quartiles. In conclusion, the prediction of clinical events by cardiac troponin T and myoglobin measured with a point-of-care analyzer in the emergency department was as good as that of the same cardiac markers and CK-MB mass measured on lab analyzers. PMID- 16214123 TI - Does measurement of oxidative damage to DNA have clinical significance? AB - Oxidative damage to DNA is the seemingly inevitable consequence of cellular metabolism. Furthermore, despite protective mechanisms, cellular levels of damage may increase under conditions of oxidative stress, arising from exposure to a variety of physical or chemical insults. Elevated levels of oxidatively damaged DNA have been measured in numerous diseases, and as a result, it has been hypothesised that such damage plays an integral role in the aetiology of that disease. This review examines the validity of this hypothesis, exploring the mechanisms by which oxidative DNA damage may lead to disease. We conclude that further validation of biomarkers of oxidative DNA damage, along with further elucidation of the role of damage in disease, may allow these biomarkers to become potentially useful clinical tools. PMID- 16214124 TI - Gold nanoparticle probes for the detection of nucleic acid targets. AB - BACKGROUND: Advances in nanoscience are having a significant impact on many scientific fields and are resulting in the development of a variety of important technologies. This impact is particularly large in the field of biodiagnostics, where a number of nanoparticle-based assays have been introduced for biomolecular detection, with DNA- or protein-functionalized gold nanoparticles used as the target-specific probes. METHODS: Assays provide an analysis of the unique biophysical properties displayed by gold nanoparticles and have advantages over conventional detection methods (e.g., molecular fluorophores, real-time polymerase chain reaction, RT-PCR, enzyme linked immunosorbent assays, ELISAs, gel electrophoresis, and microarray technologies). CONCLUSION: Some of the advantages include the assays' PCR-like sensitivity, their selectivity for target sequences, their capacity for massive multiplexing, their time efficiency, and most importantly, their ability to be performed at the point of care. PMID- 16214125 TI - A new automated colorimetric method for measuring total oxidant status. AB - OBJECTIVES: To develop a new, colorimetric and automated method for measuring total oxidation status (TOS). DESIGN AND METHODS: The assay is based on the oxidation of ferrous ion to ferric ion in the presence of various oxidant species in acidic medium and the measurement of the ferric ion by xylenol orange. The oxidation reaction of the assay was enhanced and precipitation of proteins was prevented. In addition, autoxidation of ferrous ion present in the reagent was prevented during storage. The method was applied to an automated analyzer, which was calibrated with hydrogen peroxide and the analytical performance characteristics of the assay were determined. RESULTS: There were important correlations with hydrogen peroxide, tert-butyl hydroperoxide and cumene hydroperoxide solutions (r=0.99, P<0.001 for all). In addition, the new assay presented a typical sigmoidal reaction pattern in copper-induced lipoprotein autoxidation. The novel assay is linear up to 200 micromol H2O2 Equiv./L and its precision value is lower than 3%. The lower detection limit is 1.13 micromol H2O2 Equiv./L. The reagents are stable for at least 6 months on the automated analyzer. Serum TOS level was significantly higher in patients with osteoarthritis (21.23+/-3.11 micromol H2O2 Equiv./L) than in healthy subjects (14.19+/-3.16 micromol H2O2 Equiv./L, P<0.001) and the results showed a significant negative correlation with total antioxidant capacity (TAC) (r=-0.66 P<0.01). CONCLUSIONS: This easy, stable, reliable, sensitive, inexpensive and fully automated method that is described can be used to measure total oxidant status. PMID- 16214126 TI - Gene expression during sex determination reveals a robust female genetic program at the onset of ovarian development. AB - The primary event in mammalian sexual development is the differentiation of the bipotential gonads into either testes or ovaries. Our understanding of the molecular pathways specifying gonadal differentiation is still incomplete. To identify the initial molecular changes accompanying gonadal differentiation in mice, we have performed a large-scale transcriptional analysis of XX and XY Sf1 positive gonadal cells during sex determination. In both male and female genital ridges, a robust genetic program is initiated pre-dating the first morphological changes of the differentiating gonads. Between E10.5 and E13.5, 2306 genes were expressed in a sex-specific manner in the somatic compartment of the gonads; 1223 were overexpressed in XX embryos and 1083 in XY embryos. Although sexually dimorphic genes were scattered throughout the mouse genome, we identified chromosomal regions hosting clusters of genes displaying similar expression profiles. The cyclin-dependent kinase inhibitors Cdkn1a and Cdkn1c are overexpressed in XX gonads at E11.5 and E12.5, suggesting that the increased proliferation of XY gonads relative to XX gonads may result from the overexpression of cell cycle inhibitors in the developing ovaries. These studies define the major characteristics of testicular and ovarian transcriptional programs and reveal the richness of signaling processes in differentiation of the bipotential gonads into testes and ovaries. PMID- 16214127 TI - Co-administration of fluoxetine and WAY100635 improves short-term memory function. AB - The aim of this study was to determine whether the action of the antidepressant fluoxetine or the anxiolytic buspirone could be modified by specific 5 hydroxytriptamine (5-HT(1A)) receptor blockade in a short-term memory paradigm. Male Wistar rats were trained to perform the putative short-term memory task, delayed non-matching to position. WAY100635, a selective 5-HT(1A) receptor antagonist (0.15 mg/kg), was administered 15 min before either the selective serotonin reuptake inhibitor fluoxetine (3 mg/kg), or the partial 5-HT(1A) receptor agonist and dopamine D2 receptor antagonist, buspirone (0.3 mg/kg). 8 Hydroxy-di-n-propylamino tetralin (8-OH-DPAT), a full 5-HT(1A) receptor agonist (0.3 mg/kg), was also included in the study as a positive control. WAY100635 alone had no effect on any behavioural parameter measured (response accuracy, delay lever press activity and trial completion). 8-OH-DPAT impaired response accuracy in a delay-dependent manner, an effect reversed by WAY100635. Fluoxetine also impaired response accuracy delay-dependently. WAY100635 pretreatment not only reversed this deficit but improved response accuracy, in the presence of a significant deficit in trial completion. At the dose used, buspirone showed no significant differences compared to the control group. The data suggest that fluoxetine impairs short-term memory function by the indirect activation of 5 HT(1A) receptors, but that its co-administration with WAY100635 improves short term memory function. PMID- 16214128 TI - Allosteric modulators affect the internalization of human adenosine A1 receptors. AB - To study the effect of allosteric modulators on the internalization of human adenosine A(1) receptors, the receptor was equipped with a C-terminal yellow fluorescent protein tag. The introduction of this tag did not affect the radioligand binding properties of the receptor. CHO cells stably expressing this receptor were subjected during 16 h to varying concentrations of the agonist N(6) cyclopentyladenosine (CPA) in the absence or presence of 10 microM of the allosteric enhancer PD 81,723 ((2-amino-4,5-dimethyl-3-thienyl)-[3 (trifluoromethyl)phenyl]methanone) or the allosteric inhibitor SCH-202676 (N-(2,3 diphenyl-1,2,4-thiadiazol-5(2H)-ylidene)methanamine). CPA itself was able to internalize 25% and 40% of the receptors at a concentration of 400 nM or 4 muM, respectively. Addition of either PD 81,723 or SCH-202676 alone had no effect on internalization. However, with PD 81,723 a slight amount of internalization was obtained already at 40 nM of CPA and at 400 nM CPA 59% of the receptors internalized. SCH-202676 on the other hand effectively prevented CPA-induced internalization of the receptor. PMID- 16214129 TI - Mesenchymal stem cells derived from bone marrow favor tumor cell growth in vivo. AB - Mesenchymal stem cells (MSCs) have generated a great deal of interest in clinical application because of their potential use in regenerative medicine and tissue engineering. However, the therapeutic application of MSCs still remain limited unless the favorable effect of MSCs for tumor growth in vivo and the long-term safety of the clinical applications of MSCs are better understood. In this study, MSCs derived from fetal bone marrow (FMSCs) and adult MSCs (AMSCs) alone or FMSCs and AMSCs with tumor cell line (F6 or SW480) together were transplanted subcutaneously into BALB/c-nu/nu mice to observe the outgrowth of tumor, and the characteristics of tumor cells were investigated by pathological and immunohistochemical methods, flow cytometry and real-time quantitative PCR. The results showed that both FMSCs and AMSCs could favor tumor growth in vivo. The pathologic examination revealed that tumor tissues had rich vessel distribution, extensive necrosis and invasion surrounding normal tissues, such as muscular tissue and subcutaneous tissue. In the immunohistochemical examination, tumor cells mixed with MSCs transplanted subcutaneously exhibited elevated capability of proliferation, rich angiogenesis in tumor tissues and highly metastatic ability. To understand whether MSCs affected the general properties of the tumor cells in vivo, the expression of some surface antigens and Bmi-1 gene of tumor tissue cells was detected in this study. The results indicated that these parameters were not affected after the interaction of MSCs with tumor cells in vivo. These findings suggested that MSCs could favor tumor growth in vivo. It is necessary to carry out a study for assurance of the long-term safety before MSCs were used as a therapy tools in regenerative medicine and tissue engineering. PMID- 16214130 TI - Identification of a nuclear export signal in the KSHV latent protein LANA2 mediating its export from the nucleus. AB - LANA2 is a latent protein detected in Kaposi's sarcoma-associated herpesvirus (KSHV)-infected B cells that inhibits p53-dependent transcriptional transactivation and apoptosis and PKR-dependent apoptosis, suggesting an important role in the transforming activity of the virus. It has been reported that LANA2 localizes into the nucleus of both KSHV-infected B cells and transiently transfected HeLa cells. In this study, we show that LANA2 is a nucleocytoplasmic shuttling protein that requires a Rev-type nuclear export signal located in the C-terminus to direct the protein to the cytoplasm, through an association with the export receptor CRM1. In addition, a functional protein kinase B (PKB)/Akt phosphorylation motif partially overlapping with the nuclear export signal was identified. Nuclear exclusion of LANA2 was negatively regulated by the phosphorylation of threonine 564 by Akt. The ability of LANA2 to shuttle between nucleus and cytoplasm has implications for the function of this viral protein. PMID- 16214131 TI - Insulin-like growth factor binding protein (IGFBP)-3 and IGFBP-5 mediate TGF-beta and myostatin-induced suppression of proliferation in porcine embryonic myogenic cell cultures. AB - We have previously shown that cultured porcine embryonic myogenic cells (PEMC) produce both insulin-like growth factor binding protein (IGFBP)-3 and IGFBP-5 and secrete these proteins into their media. Exogenously added recombinant porcine (rp) IGFBP-3 and rpIGFBP-5 act via IGF-dependent and IGF-independent mechanisms to suppress proliferation of PEMC cultures. Furthermore, immunoneutralization of endogenous IGFBP-3 and IGFBP-5 in the PEMC culture medium results in increased DNA synthesis rate suggesting that endogenous IGFBP-3 and IGFBP-5 suppress PEMC proliferation. TGF-beta superfamily members myostatin and TGF-beta1 have also been shown to suppress proliferation of myogenic cells, and treatment of cultured PEMC with either TGF-beta1 or myostatin significantly (P < 0.01) increases levels of IGFBP-3 and -5 mRNA. We have previously shown that immunoneutralization of IGFBP-3 decreases the proliferation-suppressing activity of TGF-beta1 and myostatin. Here, we show that immunoneutralization of IGFBP-5 also significantly (P < 0.05) decreases the DNA synthesis-suppressing activity of these molecules. Simultaneous immunoneutralization of both IGFBP-3 and IGFBP-5 in TGF-beta1 or myostatin-treated PEMC cultures restores Long-R3-IGF-I-stimulated DNA synthesis rates to 90% of the levels observed in control cultures receiving no TGF-beta1 or myostatin treatment (P < 0.05). Even though immunoneutralization of IGFBP-3 and 5 increased DNA synthesis rates in TGF-beta1 or myostatin-treated PEMC cultures, phosphosmad2 levels in these cultures were not affected. These findings strongly suggest that IGFBP-3 and IGFBP-5 affect processes downstream from receptor mediated Smad phosphorylation that facilitate the ability of TGF-beta and myostatin to suppress proliferation of PEMC. PMID- 16214132 TI - Transient attenuation of neuropathic manifestations in rats following lesion or reversible block of the lateral thalamic somatosensory nuclei. AB - BACKGROUND AND AIMS: Nociceptive behavior in animal models for mononeuropathy has been shown to be altered by spinal tract lesions which suggest a possible supraspinal modulation. The thalamus constitutes a chief center for the processing of nociception. We have, therefore, investigated the effects of transient or permanent blocks of the lateral somatosensory thalamic nuclei (the ventrobasal complex) on the neuropathic manifestations in rats. METHODS: Different groups of rats (n = 5-6) were subjected to mononeuropathy, following the spared nerve injury model, known to produce sustained heat hyperalgesia and tactile and cold allodynia which peaked about 2 weeks after nerve injury. This was followed by stereotaxic placement of either electrolytic or chemical lesions or implantation of mini osmotic pump for slow release of lidocaine in the ventrobasal complex. RESULTS: Chronic electrolytic and chemical lesions or reversible block of the lateral somatosensory thalamus produced transient (1-2 weeks) attenuation of neuropathic manifestations along with a persistent decrease of the hot plate latency. The most pronounced effect was observed on heat hyperalgesia, and the least significant and short-lived effect was observed on cold allodynia. CONCLUSION: We conclude that the lateral somatosensory thalamic complex is involved in the processing of neuropathic manifestations but cannot be considered as an obligatory or exclusive relay center for the neuropathic syndromes. PMID- 16214133 TI - Activation of phospholipase D by 8-Br-cAMP occurs through novel pathway involving Src, Ras, and ERK in human endometrial stromal cells. AB - We investigated the mechanism of 8-Br-cAMP-mediated phospholipase D (PLD) activation using a primary cell culture system of human endometrial stromal cells (ES cells). PLD activity was increased by the treatment of ES cells with 8-Br cAMP, maximally at 5 min. To determine whether the effects of 8-Br-cAMP on PLD occurred as a consequence of PKC activation, ES cells were preincubated for 15 min with RO320432 (1 microM) and GF109203X (1 microM), the PKC inhibitors, or they were pretreated for 24h with phorbol myristate acetate (100 nM) to downregulate PKC. However, these treatments had no effects on PLD activation induced by 8-Br-cAMP. Furthermore, 8-Br-cAMP had no effects on the subcellular distribution of PKC alpha and PKC betaI, confirming no involvement of PKC. 8-Br cAMP activated ERK1/2, maximally at 5 min, and PD98059 (MEK inhibitor: 50 microM) and transfection of ES cells with dominant negative (DN)-MEK completely inhibited 8-Br-cAMP-induced PLD activation, suggesting that ERK1/2 mediates the PLD activation. To investigate the involvement of protein kinase A (PKA), Src, and Ras in 8-Br-cAMP-induced PLD activation, we used PKA inhibitor, H89 and Rp-cAMPs, and transfections of DN-Src and DN-Ras. H-89 and Rp-cAMPs completely blocked 8-Br cAMP-mediated PLD and ERK activation, implying the involvement of PKA in this PLD activation. In addition, transfection of ES cells with DN-Src, or DN-Ras partially inhibited 8-Br-cAMP-induced ERK1/2 and consequently PLD activation, whereas cotransfection of DN-Src and DN-Ras completely inhibited ERK1/2 and PLD activation, suggesting that Src and Ras independently regulate ERK/PLD activation. Taken together, these results demonstrate a novel pathway in ES cells that 8-Br-cAMP activate PLD through PKA and ERK1/2 and this ERK/PLD activation by 8-Br-cAMP is mediated by Src and Ras, separately. PMID- 16214134 TI - Prediction and validation of microRNAs and their targets. AB - MicroRNAs are short non-coding RNAs that inhibit translation of target genes by binding to their mRNAs, and have been shown to play a central role in gene regulation in health and disease. Sophisticated computer-based prediction approaches of microRNAs and of their targets, and effective biological validation techniques for validating these predictions, now play a central role in discovery of microRNAs and elucidating their functions. PMID- 16214135 TI - Inhibition of neuronal nitric-oxide synthase by phosphorylation at Threonine1296 in NG108-15 neuronal cells. AB - We demonstrate that neuronal nitric-oxide synthase (nNOS) is directly inhibited through the phosphorylation of Thr(1296) in NG108-15 neuronal cells. Treatment of NG108-15 cells expressing nNOS with calyculin A, an inhibitor of protein phosphatase 1 and 2A, revealed a dose-dependent inhibition of nNOS enzyme activity with concomitant phosphorylation of Thr(1296) residue. Cells expressing a phosphorylation-deficient mutant in which Thr(1296) was changed to Ala proved resistant to phosphorylation and suppression of NOS activity. Mimicking phosphorylation mutant of nNOS in which Thr(1296) is changed to Asp showed a significant decrease in nNOS enzyme activity, being competitive with NADPH, relative to the wild-type enzyme. These data suggest that phosphorylation of nNOS at Thr(1296) may involve the attenuation of nitric oxide production in neuronal cells through the decrease of NADPH-binding to the enzyme. PMID- 16214136 TI - The dimeric form of flavocytochrome P450 BM3 is catalytically functional as a fatty acid hydroxylase. AB - In the model P450 BM3 system, the P450 is fused to its diflavin reductase partner in a single polypeptide. BM3 dimerizes in solution, but the catalytic relevance of the phenomenon was hitherto unknown. We show that BM3 fatty acid hydroxylase specific activity decreases sharply at low enzyme concentrations, consistent with separation of active dimer into inactive monomer. Reductase-dependent specific activities are maintained or enhanced at low concentration, suggesting inter flavin electron transfer is unaffected. Fatty acid oxidation is reconstituted by mixing inactive oxygenase (A264H) and FMN-depleted (G570D) mutants, demonstrating that inter-monomer (FMN(1)-to-heme(2)) electron transfer supports oxygenase activity in the BM3 dimer. PMID- 16214137 TI - Selective activation of adrenergic beta1 receptors induces heme oxygenase 1 production in RAW264.7 cells. AB - We hypothesized that catecholamines through beta-adrenoceptor might modulate macrophage function. We showed that isoproterenol concentration-dependently induced HO-1 production through beta(1)-but not beta(2)-adrenoceptor. Production was increased by forskolin and inhibited by pretreatment with the PKA inhibitor, H-89. Furthermore, induction of HO-1 by isoproterenol effectively protected RAW264.7 cells from effects of glucose oxidase treatment, which was abrogated either by HO-1 inhibitor, ZnPP IX and beta-adenoceptor antagonist, propranolol. Thus, stimulation of HO-1 production through beta(1)-adenoceptors, and via the PKA pathways by isoproterenol, can enable RAW264.7 cells to resist oxidant stress, suggesting that catecholamine hormones may be necessary, at least, to maximize defending role of macrophages. PMID- 16214138 TI - The dimer interface of the SARS coronavirus nucleocapsid protein adapts a porcine respiratory and reproductive syndrome virus-like structure. AB - We have employed NMR to investigate the structure of SARS coronavirus nucleocapsid protein dimer. We found that the secondary structure of the dimerization domain consists of five alpha helices and a beta-hairpin. The dimer interface consists of a continuous four-stranded beta-sheet superposed by two long alpha helices, reminiscent of that found in the nucleocapsid protein of porcine respiratory and reproductive syndrome virus. Extensive hydrogen bond formation between the two hairpins and hydrophobic interactions between the beta sheet and the alpha helices render the interface highly stable. Sequence alignment suggests that other coronavirus may share the same structural topology. PMID- 16214139 TI - Dicing and slicing: the core machinery of the RNA interference pathway. AB - RNA interference (RNAi) is broadly defined as a gene silencing pathway that is triggered by double-stranded RNA (dsRNA). Many variations have been described on this theme. The dsRNA trigger can be supplied exogenously, as an experimental tool, or can derive from the genome in the form of microRNAs. Gene silencing can be the result of nucleolytic degradation of the mRNA, or by translational suppression. At the heart of the pathway are two ribonuclease machines. The ribonuclease III enzyme Dicer initiates the RNAi pathway by generating the active short interfering RNA trigger. Silencing is effected by the RNA-induced silencing complex and its RNaseH core enzyme Argonaute. This review describes the discovery of these machines and discusses future lines of work on this amazing biochemical pathway. PMID- 16214140 TI - A eukaryotic carboxyl-terminal signal sequence translocating large hydrophilic domains across membranes. AB - Yeast Golgi ecto-ATPase Ynd1p is an unusual type III membrane protein with the longest translocated N-terminus reported. Sequential deletion analysis reveals that translocation of this 500-residue-long hydrophilic domain across the membranes requires the C-terminal transmembrane domain of Ynd1p and its flanking regions. Additional studies indicate that the topogenic sequence of Ynd1p overrides the effect of a reverse signal-anchor sequence present at the N terminus of Ynd1p, while it is not affected by a classic signal sequence at the N terminus. When placed at the C-terminal end, the sequence can translocate large extracellular domains of two membrane proteins across the membranes. The data demonstrate the existence of a true eukaryotic C-terminal signal sequence. PMID- 16214141 TI - The effects of partial thiamin deficiency and oxidative stress (i.e., glyoxal and methylglyoxal) on the levels of alpha-oxoaldehyde plasma protein adducts in Fischer 344 rats. AB - We hypothesized that in marginal thiamin deficiency intracellular alpha oxoaldehydes form macromolecular adducts that could possibly be genotoxic in colon cells; and that in the presence of oxidative stress these effects are augmented because of decreased detoxification of these aldehydes. We have demonstrated that reduced dietary thiamin in F344 rats decreased transketolase activity and increased alpha-oxoaldehyde adduct levels. The methylglyoxal protein adduct level was not affected by oral glyoxal or methylglyoxal in the animals receiving thiamin at the control levels but was markedly increased in the animals on a thiamin-reduced diet. These observations are consistent with our suggestion that the induction of aberrant crypt foci with marginally thiamin-deficient diets may be a consequence of the formation of methylglyoxal adducts. PMID- 16214142 TI - A cDNA for a putative type III deiodinase in the trout (Oncorhynchus mykiss): influence of holding conditions and thyroid hormone treatment on its hepatic expression. AB - A putative rainbow trout type III deiodinase (D3) cDNA was amplified by PCR, using primers to evolutionarily conserved sequences. The RACE-derived complete cDNA was then identified by sequence comparison to that in tilapia and other vertebrates. The cDNA coded for a predicted 31,500 kDa protein of 278 amino acids, with a hydrophobic trans-membrane segment and with 80% similarity to tilapia D3 and 39% similarity to rainbow trout type II deiodinase (D2). It also showed a selenocysteine codon at position 141 and a putative SECIS element in the 3' untranslated end. In the liver, a second form of D3 was found that differed only at this 3' untranslated region; the coding region was identical in both forms. The D3 mRNA, measured by RT-PCR using primers located within the common, translated portion of the cDNA, was expressed in the brain and, depending on thyroidal status, in liver and kidney. Holding trout for 7 days in static water as opposed to flowing water caused increased plasma T4 levels, decreased hepatic D2 mRNA levels and T4 outer-ring deiodination (ORD) activity and increased D3 mRNA levels and T3 inner-ring (IRD) activity. Trout held in flowing water and fed T3 for 7 days showed increased plasma T3 levels and hepatic D3 mRNA levels and T3 IRD activity but decreased D2 mRNA levels and T4ORD activity. Trout held in static water and exposed to ambient T4 for 7 days showed increased plasma T4 levels and hepatic T3IRD activity but with no significant change in D2 or D3 mRNA levels. We conclude that hepatic D3 mRNA levels and T3IRD activity are enhanced and D2 mRNA levels and T4ORD activity are suppressed by adverse holding conditions or T3 treatment suggesting that the putative D3 cDNA and D2 cDNA represent respectively the genes determining T3IRD and T4ORD activities. However, there were changes in the ratios of mRNA levels to enzyme activity, raising the potential for post-transcriptional regulation and showing that mRNA levels alone may be unreliable indices of deiodinase activity. Post-transcriptional regulation of D3 enzyme activity may be influenced by the observed alternative 3' and 5' untranslated regions of the D3 mRNA. PMID- 16214143 TI - Receptors for neuropeptide Y, gamma-aminobutyric acid and dopamine differentially regulate Ca2+ currents in Xenopus melanotrope cells via the G(i) protein beta/gamma-subunit. AB - Secretion of alpha-melanophore-stimulating hormone (alphaMSH) from pituitary melanotrope cells of the amphibian Xenopus laevis is under inhibitory synaptic control by three neurotransmitters produced by the suprachiasmatic nucleus: gamma aminobutyric acid (GABA), neuropeptide Y (NPY) and dopamine (DA). These inhibitory effects occur through G(i)-protein-coupled receptors (G(i)PCR), and differ in strength: GABA(B)-receptor-induced inhibition is the weakest, whereas DA (via a D2-receptor) and NPY (via a Y1-receptor) strongly inhibit, with NPY having a long-lasting effect. Previously it was shown that DA inhibits two (R- and N-type channel) of the four voltage-operated Ca2+ channels in the melanotrope, and that only part of this inhibition is mediated by beta/gamma subunits of the G(i) protein. We here demonstrate that also the Y1- and GABA(B) receptor inhibit only part of the total Ca2+ current (I(Ca)), with fast activation and inactivation kinetics. However, GABA(B)-mediated inhibition is weaker than the inhibitions induced via Y1- and D2-receptors (-21 versus -27% and -30%, respectively). Using a depolarizing pre-pulse protocol it was demonstrated that GABA(B)-induced inhibition of I(Ca) most likely depends on Gbeta/gamma subunit activation whereas Y1- and D2- induced inhibitions are only partially mediated by Gbeta/gamma-subunits. No differences were found between the Y1- and D2-induced inhibitions. These results imply that activation of different G(i)PCR inhibits the I(Ca) through different mechanisms, a phenomenon that may underlie the different potencies of the suprachiasmatic neurotransmitters to inhibit alphaMSH secretion. PMID- 16214145 TI - Prenatal diagnosis and molecular analysis of type 1 thanatophoric dysplasia. PMID- 16214144 TI - Prognostic value of umbilical-middle cerebral artery pulsatility index ratio in fetuses with growth restriction. AB - OBJECTIVE: To study the utility of Doppler velocimetry and computerized cardiotocography in the management of intrauterine growth restriction and prediction of neonatal outcome. PATIENTS AND METHODS: Seventy-two pregnant women with fetuses showing growth restriction and delivered within 48 h of their last Doppler velocimetry evaluation. The last computerized cardiotocographic trace from these fetuses was used for statistical analysis, and the last trace from the healthy fetuses of 93 consecutive women undergoing cesarean section was used as control. Umbilical artery pulsatility index (UA PI), middle cerebral artery pulsatility index (MCA PI), UA PI/MCA PI ratio, and uterine artery resistance index (Ut RI) were assessed. RESULTS: Among women with growth-restricted fetuses, all parameters were significantly higher in those who had hypertension; and in those who had diabetes, only the UA PI/MCA ratio was significantly higher. Umbilical artery PI values and the UA PI/MCA ratio were higher in those who had a nonreassuring result to computerized nonstress test immediately before delivery. A multiple logistic analysis showed that the UA PI/MCA ratio was the only Doppler velocimetry parameter predicting cardiotocographic nonreactivity; furthermore, the predictivity of extended newborn hospitalization (longer than 15 days) was verified, with a sensitivity of 56% and a specificity of 92% when the ratio was higher than 1.26. CONCLUSION: The MCA PI of fetuses with growth restriction should be assessed. The UA PI/MCA ratio is predictive of a nonreactive computerized cardiotocography trace and of prolonged neonatal hospitalization. PMID- 16214146 TI - Involvement of nurses in caring for patients requesting euthanasia in Flanders (Belgium): a qualitative study. AB - BACKGROUND: Although nurses worldwide are confronted with euthanasia requests, how nurses experience their involvement in euthanasia remains unclear. OBJECTIVES: To explore nurses' involvement in the care for patients requesting euthanasia. DESIGN: A qualitative grounded theory strategy. SETTING: Two general hospitals (A, B) and a palliative care setting in Flanders (Belgium). PARTICIPANTS: Nurses who fulfilled the following inclusion criteria: (a) Dutch speaking; (b) working for at least one year in hospital A or B; (c) working at least part-time (50%); and (d) ever received a euthanasia request. We collected data using purposeful sampling, superseded by theoretical sampling in a palliative care setting. The sample included one intensive care nurse, one oncology nurse, eight palliative care nurses, and five internal medicine nurses. All but five were women. Their age ranged from 24 to 49 years. METHODS: We conducted one-on-one semi-structured interviews between November 2001 and September 2002. Grounded theory was applied for guiding data collection and analysis. The trustworthiness of data was ensured by several strategies. RESULTS: Although euthanasia was still illegal, the nurses unanimously stated that they had an important role in caring for patients requesting euthanasia. Their personal and intense involvement caused them to experience a spectrum of emotions, chief among them being a sense of powerlessness. Several elements contributed to the nurses' conflicted involvement. Nurses became frustrated if the context (e.g., lack of time) hindered their efforts to provide compassionate care. The palliative care setting and its associated culture (group mentality, care philosophy) created the opportunity for nurses to take time to holistically support patients and their relatives. CONCLUSIONS: Hospital nurses are confronted with patients' euthanasia requests. Each stage of this process requires that the nurses possess specific competencies. Their willingness to personally care for these patients, in addition to their specific care expertise, allows them to be skilled companions. PMID- 16214147 TI - The TNF alpha converting enzyme (TACE/ADAM17) is expressed in the atherosclerotic lesions of apolipoprotein E-deficient mice: possible contribution to elevated plasma levels of soluble TNF alpha receptors. AB - TNF alpha converting enzyme (TACE) critically regulates the inflammatory processes as it releases from the cell surface several transmembrane proteins, including TNFalpha (TNF) and its receptors TNFR1 and TNFR2. We investigated the expression of TACE in atherosclerotic lesions of apolipoproteinE-deficient (apoE (-/-)) mice. Five-week-old apoE(-/-) male mice were fed a high-fat diet and examined at 5, 10, 15 and 25 weeks of age. A group of wild-type C57BL/6 mice (WT) fed the high-fat diet for 25 weeks was included. In apoE(-/-) mice, lesions progressed with time in both aortic sinus and arch, in which TACE immunostaining also increased particularly between 5 and 15 weeks. TACE expression was also observed in human atherosclerotic plaques. The plasma levels of soluble TNFR1 and TNFR2 rose with atherosclerosis. In the 25-week-old WT mice, no lesions were observed and the plasma levels of TNFRs were 17% of those of age-matched apoE(-/ ) mice. Incubated aortas of 25-week-old apoE(-/-) mice released much higher amounts of sTNF and sTNFRs than did aortas of 5-week-old apoE(-/-) mice or 25 week-old WT mice. Active TACE was expressed at the surface of macrophages isolated from apoE(-/-) mice. In conclusion, TACE expression is associated with lesions in atherosclerosis-prone sites. Our data suggest that atherosclerotic lesions-expressing TACE may contribute to the elevated levels of circulating sTNFRs. PMID- 16214148 TI - Hyperinsulinemia is an independent predictor for complex atherosclerotic lesion of thoracic aorta in non-diabetic patients. AB - OBJECTIVE: Hyperinsulinemia is a well known risk factor for cardiovascular event. However, it is not known whether hyperinsulinemia facilitates atherosclerotic complex lesions of aorta in non-diabetic patients. We investigated whether hyperinsulinemia is an independent marker of severity of atherosclerosis in thoracic aorta of non-diabetic patients using multiplane transesophageal echocardiography (TEE). RESEARCH DESIGN AND METHODS: Non-diabetic 90 patients with cardiovascular disease underwent TEE, and were analyzed for plasma insulin levels of oral glucose tolerance test, conventional atherosclerotic risk factors and coronary angiographic features. RESULTS: Thoracic aortic plaques were detected in 84 patients (93%). The complex atherosclerotic lesions were observed in 35 (39%) patients, most frequently at the part of aortic arch (p<0.005), showing the greatest atheroma score in thoracic aorta (p<0.05). Univariate analysis showed age, male gender, smoking, coronary artery disease, HDL cholesterol, insulin levels in glucose tolerance test and homeostasis model assessment insulin resistance index (HOMA index) were found to be significant predictors of complex atherosclerotic lesions. Multivariate regression analysis revealed that HOMA index was an independent predictor of complex atherosclerotic lesions (odds ratio 1.93, p=0.006). There was a significant positive correlation between HOMA index and the atheroma score of thoracic aorta (p<0.001). CONCLUSIONS: Hyperinsulinemia is an independent predictor of complex atherosclerotic lesions detected by TEE in the thoracic aorta of non-diabetic patients. PMID- 16214149 TI - Critical role of bradykinin-eNOS and oxidative stress-LOX-1 pathway in cardiovascular remodeling under chronic angiotensin-converting enzyme inhibition. AB - To elucidate the molecular mechanisms of the cardioprotective effect of angiotensin-converting enzyme (ACE) inhibitors, we evaluated whether the effect of quinapril involved in bradykinin-endothelial nitric oxide synthase (eNOS) and oxidative stress-lectin-like oxidized LDL receptor-1 (LOX-1) pathway. Dahl salt sensitive hypertensive (DS) rats were fed a diet containing 8% NaCl and treated with one of the following drug combinations for 5 weeks, from 6 weeks of age to left ventricular hypertrophy stage (11 weeks): vehicle; quinapril; quinapril plus the bradykinin B2 receptor antagonist FR172357; the NAD(P)H oxidase inhibitor apocynin; or quinapril plus apocynin. eNOS expression, which was decreased in hypertrophy stage, was significantly increased by quinapril and/or apocynin, but not by quinapril plus FR172357. Upregulated expression of NAD(P)H oxidase p22phox, p47phox, gp91phox and LOX-1 was significantly decreased by quinapril to a similar degree as after treatment with apocynin, but not by quinapril plus FR172357. Quinapril and/or apocynin treatment effectively ameliorated left ventricular weight and vascular changes such as increase in medial thickness and perivascular fibrosis and suppressed expression of transforming growth factor beta1, type I collagen and fibronectin mRNA, but not that of quinapril plus FR172357. These results suggest that the ACE inhibitor quinapril may have cardioprotective effects in this model of hypertension mediated at least in part through effects on the bradykinin-eNOS and oxidative stress-LOX-1 pathway. PMID- 16214150 TI - Nonlinear finite element simulation to elucidate the efficacy of slit arteriotomy for end-to-side arterial anastomosis in microsurgery. AB - The slit arteriotomy for end-to-side arterial microanastomosis is a technique used to revascularize free flaps in reconstructive surgery. Does a slit open to a width sufficient for blood supply? How is the slit opening affected by factors, such as arterial wall thickness and material stiffness? To answer these questions we propose a nonlinear finite element procedure to simulate the operation. Through modeling the arteries using hyper-elastic shell elements, our simulation reveals that the slit opens up to a width even larger than the original diameter of the donor artery, allowing sufficient blood supply. It also identifies two factors that explain the opening of the slit: blood pressure which is predominant in most cases, and the forces applied to the slit by the donor artery. During simulation, when we increase the donor artery thickness and stiffness, it is found that the contribution of blood pressure to the slit opening decreases while that of the forces applied by the donor artery increases. This result indicates that sometimes the forces applied by the donor artery can play an even more significant role than the blood pressure factor. Our simulation elucidates the efficacy of the slit arteriotomy. It improves our understanding of the interplay between blood pressure and donor vessel factors in keeping the slit open. PMID- 16214152 TI - Effects of administration of transforming growth factor (TGF)-beta1 and anti-TGF beta1 antibody on the mechanical properties of the stress-shielded patellar tendon. AB - Previous studies have shown that, in the stress-shielded patellar tendon, the mechanical properties of the tendon were dramatically reduced and TGF-beta was over-expressed in tendon fibroblasts. In the present study, therefore, we tested two supportive hypotheses using 40 rabbits: One was that an application of TGF beta1 might significantly increase the tensile strength and the tangent modulus of the stress-shielded patellar tendon. The other one was that an administration of anti-TGF-beta1 antibody might significantly reduce the mechanical properties of the stress-shielded patellar tendon. In the results, an application of 4-ng TGF-beta1 significantly increased the tangent modulus of the stress-shielded patellar tendon at 3 weeks (p = 0.019), compared with the sham treatment. Concerning the tensile strength, the 4-ng TGF-beta1 application increased the average value, but a statistical significance was not reached. An application of 50-microg anti-TGF-beta1 antibody significantly reduced the tangent modulus and the tensile strength of the stress-shielded patellar tendon at 3 weeks (p = 0.0068 and p = 0.0355), compared with the sham treatment. Because the stress shielding treatment used in this study dramatically reduces the tangent modulus and the tensile strength of the patellar tendon, the present study suggested that an administration of TGF-beta1 weakly but significantly inhibited the reduction of the mechanical properties of the stress-shielded patellar tendon, and that inactivation of TGF-beta1 with its antibody significantly enhanced the reduction of the mechanical properties that occurs in the stress-shielded patellar tendon. These results suggested that TGF-beta1 plays an important role in remodeling of the stress-shielded patellar tendon. PMID- 16214153 TI - A novel microstructural approach in tendon viscoelastic modelling at the fibrillar level. AB - Novel applications in rehabilitation, surgery and tissue engineering require the knowledge of the mechanical behaviour of the tissues at microstructural level. The aim of this work is to investigate the viscoelastic properties of the tendon from the interaction of its biological constituents in the fibrillar network. Traction, relaxation and creep in-vitro tests have been performed on porcine flexor digital tendons. A viscoelastic constitutive equation at finite deformation is presented. The fibrillar deformation modes are described through a network of adaptive links between collagen type I and decorin. The theoretical predictions fit accurately the experimental data. The results of the model demonstrate the mechanical importance of glycosaminoglycan chains of decorin for the differential recruitment and the activation of fibrillar collagen. PMID- 16214154 TI - Modeling of weak blast wave propagation in the lung. AB - Blast injuries of the lung are the most life-threatening after an explosion. The choice of physical parameters responsible for trauma is important to understand its mechanism. We developed a one-dimensional linear model of an elastic wave propagation in foam-like pulmonary parenchyma to identify the possible cause of edema due to the impact load. The model demonstrates different injury localizations for free and rigid boundary conditions. The following parameters were considered: strain, velocity, pressure in the medium and stresses in structural elements, energy dissipation, parameter of viscous criterion. Maximum underpressure is the most suitable wave parameter to be the criterion for edema formation in a rabbit lung. We supposed that observed scattering of experimental data on edema severity is induced by the physiological variety of rabbit lungs. The criterion and the model explain this scattering. The model outlines the demands for experimental data to make an unambiguous choice of physical parameters responsible for lung trauma due to impact load. PMID- 16214155 TI - Trabecular architecture can remain intact for both disuse and overload enhanced resorption characteristics. AB - The paradigm that bone metabolic processes are controlled by osteocyte signals have been the subject of investigation in many recent studies. One hypothesis is that osteoblast formation is enhanced by these signals, and that osteoclast resorption is enhanced by the lack of them. Reduced, or absent, osteocyte signaling can be an effect of reduced mechanical loading (disuse) or of defects in the canalicular network, due to microcracks. This would mean that bone is resorbed precisely there where it is mostly needed. In our study, we addressed this apparent contradiction. The purpose was to investigate how alternative strain-based local stimuli for osteoclasts to resorb bone would affect remodeling and adaptation of the trabecular architecture. For this purpose, a computer simulation model was used, which couples morphological and mechanical effects of local bone metabolism to changes in trabecular architecture and density at large. Six resorption characteristics were studied in the model: (I) resorption occurs spatially random, (II) resorption is enhanced or (III) strongly enhanced where there is disuse, (IV) resorption is enhanced or (V) strongly enhanced where there are high strains, i.e. overload, and (VI) resorption is enhanced where there is disuse and where there are high strains. Results showed that the rates of structural adaptation to alternative loading were higher for disuse-controlled resorption than for overload-controlled resorption. Architecture and mass remained stable for all cases except (V) in which the structure deteriorated as in osteoporotic bone. We conclude that, given the potential of osteoblasts to form bone in highly strained areas, based on signals from osteocytes, osteoclast resorption can normally be compensated for. PMID- 16214156 TI - High-throughput analysis of tetracycline and penicillin antibiotics in animal tissues using electrospray tandem mass spectrometry with selected reaction monitoring transition. AB - A simple, rapid, and simultaneous analysis method for oxytertracycline, tetracycline, chlortetracycline, penicillin G, ampicillin, and nafcillin in meat has been developed by using electrospray ionization tandem mass spectrometry. The sample preparation was performed by homogenizing with water followed by a centrifugal ultrafiltration, after addition of internal standards (demeclocycline, penicillin G-d5, ampicillin-d5 and nafcillin-d6). The MS/MS analysis involves the combined use of sample enrichment on the short column and a multiple reaction monitoring technique. The overall recoveries from animal (bovine and swine) muscle, kidney, and liver fortified at the levels of 0.05 and 0.1 ppm ranged from 70 to 115% with the coefficients of variation ranging from 0.7 to 14.8% (n = 5). Analysis time, including sample preparation and determination, is only 3h per eight sample and detection limits for all antibiotics are 0.002 ppm. The method is considered to be satisfactory for the rapid screening of the tetracycline and penicillin antibiotic residues in meat. PMID- 16214157 TI - Selectivity in preparative separations of inorganic electrolytes by size exclusion chromatography on hypercrosslinked polystyrene and microporous carbons. AB - Preparative-scale separation of concentrated solutions of simplest mineral electrolytes by size-exclusion chromatography was performed on three samples of commercially available microporous hypercrosslinked polystyrene sorbents "Macronet Hypersol" and two experimental samples of activated carbons. Selectivity of separation of a pair of electrolytes was found to be determined by the largest ions in each pair. Fortunately, selectivity rises at higher concentrations of electrolytes, which was explained by exclusion of smaller species from the concentrated solution, i.e., mobile phase, into small pores of the column packing that are inaccessible to large species. The separation of concentrated mixtures revealed another remarkable advantage of the new process - self-concentrating of each of two separated components in the corresponding fractions. Self-concentration is more pronounced for the minor component that occupied less space in the initial mixture. The new method may prove productive in processing pickle bath solutions. PMID- 16214158 TI - Highly-ordered layered organo-mineral materials prepared via reactions of n alkylphosphonic acids with apatite. AB - This work describes a novel class of layered organo-mineral materials manufactured via a single-step solution-phase reaction of n-alkylphosphonic acids (CnH(2n+1)P(O)(OH)2) with calcium hydroxyapatite mineral (CaHAP). TEM, SAXS, WAXS, FTIR, and Vapor Phase Adsorption data suggest that these alkyl-CaHAP materials present a surface-modified CaHAP matrix coated with ordered layers of calcium alkylphosphonates that are strongly adhered to the surface. Interlayer spacing increases from 1.47 (C3-CaHAP) to 4.77 nm (C18-CaHAP). According to FTIR, ordering of alkyl chains improves with the alkyl chain length. The organic loads in these alkyl-CaHAP can be controlled over a wide range (up to approximately 60%) by varying alkyl chain and the concentration of alkylphosphonic acids in the solution. PMID- 16214159 TI - The use of acridine orange base (AOB) as molecular probe to characterize nonaqueous AOT reverse micelles. AB - The behavior of acridine orange base (AOB) in nonaqueous reverse micelles composed of n-heptane/AOT/polar solvent has been performed. Ethylene glycol (EG), propylene glycol (PG), glycerol (GY), formamide (FA), dimethylformamide (DMF), and dimethylacetamide (DMA) were employed as water substitutes. The studies were performed by static and time-resolved emission spectroscopy. Thus, the distribution of AOB between the two pseudophases of the aggregates was quantified by measuring the partition constants from emission spectra at different surfactant concentration. Similar values to those obtained by means of absorption spectroscopy were obtained. This match is indicating that AOB is not experiencing partition during the lifetime of the excited state. Partitioning to the micelles is strongly favored in micelles containing hydrogen-bond donor (HBD) solvents rather than non-HBD solvents. Variations of fluorescence lifetimes with AOT concentration confirm these results. By the solvatochromic behavior of AOB in the different systems it is shown that the microenvironment at the interface is distinct from that of the bulk polar solvent, indicating that the probe senses no "free" solvent. The steady state anisotropy (r) was measured for EG/AOT/n-heptane and DMF/AOT/n-heptane systems as representatives for HBD and non-HBD polar solvents, respectively. The value of r is higher in the micelles containing EG than that obtained with DMF, and increases with AOT concentration. This is explained as due to highly structured polar solvents in the inner core. EG is interacting with the polar heads of AOT through hydrogen-bond interaction, while DMF can only interact with the Na+ counterions. This is confirmed by the time resolved emission spectra (TRES) of the probe in the micellar systems, in comparison with the bulk solvents. PMID- 16214160 TI - Interfacial characteristics of heterogeneous layers of linear polyvinylamine and branched polyethyleneimine or polyethyleneimine grafted with C12-C22 alkyl chains. AB - We investigated the characteristics of heterogeneous layers composed of linear hydrolyzed polyvinylamine and branched polyethyleneimine adsorbed at silica/water interfaces. The studies also included heterogeneous layers where branched polyethyleneimine was replaced by polyethyleneimine modified by grafting with C12 C22 alkyl chains. Surface area exclusion chromatography was used to determine the interfacial relaxation and surface affinity of the polymer molecules within homogeneous layers. The relaxation of bare and grafted polyethyleneimine was found to be small and of equal extent but to develop at different rates. Comparatively, the relaxation of hydrolyzed polyvinylamine was faster and of greater extent. Within heterogeneous layers composed of polyvinylamine and bare or grafted polyethyleneimine, the relaxation of the different molecules was strongly increased as compared to that prevailing in homogeneous layers. The chromatographic method was then used to determine the mode of layer establishment. The polymer coating profiles on successive glass fiber filters were found to depend on the sequence of injection of the two polymers, due to the interfacial stability or instability of the initially established layer. It was shown that a previously established extremely thin layer of bare or grafted polyethyleneimine molecules strongly modified the adsorption profile of subsequently adsorbed polyvinylamine molecules. PMID- 16214161 TI - Detection of multiple viruses in queens of the honey bee Apis mellifera L. AB - Individual honey bee Apis mellifera L. queens were examined for the presence of six honey bee viruses including acute bee paralysis virus, chronic bee paralysis virus, black queen cell virus, deformed wing virus, Kashmir bee virus, and sacbrood virus. All viruses, except ABPV, were detected in the samples. Among queens examined for virus infections, 93% had multiple virus infections. The detection of viruses in queens raises the possibility of a vertical transmission pathway wherein infected queens can pass virus through their eggs to their offspring. PMID- 16214162 TI - Physiological host specificity: a model using the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae) and microsporidia of row crop and other stalk-boring hosts. AB - We investigated vertical and horizontal transmission as means by which entomopathogenic microsporidia may be isolated in their hosts. Ostrinia nubilalis larvae were challenged with microsporidia isolated from other stalk-boring and row crop Lepidoptera and were susceptible to seven species. Two species were horizontally transmitted. A Nosema sp. from Eoreuma loftini was transmitted among O. nubilalis larvae but not among larvae of the E. loftini host. This species was also vertically transmitted to the offspring of infected O. nubilalis females. An rDNA sequence showed the E. loftini isolate to be Nosema pyrausta, a naturally occurring species in O. nubilalis. Our results suggest that both horizontal and vertical transmission provide physiological barriers to host switching in the microsporidia, thus restricting the natural host range. PMID- 16214163 TI - Bacillus cereus produces several nonproteinaceous insecticidal exotoxins. AB - Bacillus cereus is mainly known as a human food-borne opportunistic pathogen. Here, we used biological assays and HPLC to investigate the ability of B. cereus to produce insecticidal exotoxins during the stationary growth phase. None of the 575 B. cereus strains screened produced detectable levels of beta-exotoxin I, a small, heat-stable insecticidal nucleotide analogue. However, six out of a subset of 270 B. cereus strains produced several small, nonproteinaceous insecticidal exotoxins different from beta-exotoxin I. Thus, B. cereus can secrete a large array of proteinaceous and nonproteinaceous toxins acting on insects and mammals. PMID- 16214164 TI - PCR diagnostic methods for Ascosphaera infections in bees. AB - Fungi in the genus Ascosphaera are the causative agents of chalkbrood, a major disease affecting bee larval viability. Identification of individual Ascosphaera species based on morphological features has been difficult due to a lack of distinguishing characteristics. Most identifications are based on the size and shape of the ascomata, spore balls and conidia. Unfortunately, much overlap occurs in the size of these structures, and some Ascosphaera species will not produce sexual structures in vitro. We report a quick and reliable diagnostic method for identifying Ascosphaera infections in Megachile bees (leafcutting bees) using PCR markers that employ genus-specific primers for Ascosphaera, and species-specific primers for species known to be associated with Megachile spp. Using these methods, species identifications can be performed directly on bees, including asymptomatic individuals. Furthermore, the PCR markers can detect co infections of multiple Ascosphaera species in a single host. We also identified a marker for Ascosphaera apis, the predominant cause of chalkbrood in Apis mellifera, the honey bee. Our diagnostic methods eliminate the need for culturing samples, and could be used to process a large number of field collected bee larvae. PMID- 16214165 TI - Urosporidium sp. hyperparasite of the turbellarian Paravortex cardii in the cockle Cerastoderma edule. AB - Cysts with spores showing different degree of maturity and a single plasmodium were observed in the connective tissue of the turbellarian Paravortex cardii located in the digestive lumen of the cockle Cerastoderma edule. The study of spore morphology by transmission electron microscopy revealed that they correspond to an haplosporidian belonging to the genus Urosporidium. Spore ornaments were similar to those described from Urosporidium spisuli, infecting a nematode parasite of the Atlantic surf clam, Spisula solidissima. PMID- 16214166 TI - Disordered p27Kip1 exhibits intrinsic structure resembling the Cdk2/cyclin A bound conformation. AB - p27Kip1 (p27) influences cell division by regulating nuclear cyclin-dependent kinases. Before binding, p27 is at least partially disordered and folds upon binding its Cdk/cyclin targets. 30-40% of human proteins, including p27, are predicted to contain disordered segments, and have been termed intrinsically unstructured proteins (IUPs). Unfortunately, the inherent dynamics of IUPs hamper detailed analysis of their structure/function relationships. Here, we describe the use of molecular dynamics (MD) computations and solution NMR spectroscopy to reveal that several segments of the p27 kinase inhibitory domain (p27-KID), in addition to the previously characterized helical segment, exist as highly populated, intrinsically folded structural units (IFSUs). Several IFSUs resemble structural features of bound p27-KID, while another exhibits alternative conformations. Interestingly, the highly conserved, specificity determining segment of p27 is shown to be highly disordered. Elucidation of IFSUs within p27 KID allows consideration of their influences on the thermodynamics and kinetics of Cdk/cyclin binding. The degree to which IFSUs are populated within p27-KID is surprising and suggests that other putative IUPs contain IFSUs that may be studied using similar techniques. PMID- 16214167 TI - RNA tertiary interactions mediate native collapse of a bacterial group I ribozyme. AB - Large RNAs collapse into compact intermediates in the presence of counterions before folding to the native state. We previously found that collapse of a bacterial group I ribozyme correlates with the formation of helices within the ribozyme core, but occurs at Mg2+ concentrations too low to support stable tertiary structure and catalytic activity. Here, using small-angle X-ray scattering, we show that Mg2+-induced collapse is a cooperative folding transition that can be fit by a two-state model. The Mg2+ dependence of collapse is similar to the Mg2+ dependence of helix assembly measured by partial ribonuclease T1 digestion and of an unfolding transition measured by UV hypochromicity. The correspondence between multiple probes of RNA structure further supports a two-state model. A mutation that disrupts tertiary contacts between the L9 tetraloop and its helical receptor destabilized the compact state by 0.8 kcal/mol, while mutations in the central triplex were less destabilizing. These results show that native tertiary interactions stabilize the compact folding intermediates under conditions in which the RNA backbone remains accessible to solvent. PMID- 16214168 TI - The high-mobility group transcription factor Sox10 interacts with the N-myc interacting protein Nmi. AB - The high-mobility group transcription factor Sox10 exerts many different roles during development of the neural crest and nervous system. To unravel its complex transcriptional functions, we have started to look for interaction partners. Here, we identify an association of Sox10 with the N-myc interactor Nmi, which was mediated by the high-mobility group of Sox10 and the central region of Nmi. In vivo relevance of this interaction is indicated by the fact that both proteins were co-expressed in glial cells, gliomas and in the spinal cord. Additionally, subcellular localization of Nmi in C6 glioma depended on the presence of Sox10 such that nuclear Nmi was more frequent in Sox10-expressing cells. Importantly, Nmi modulated the transcriptional activity of Sox10 in reporter gene assays. Nmi effects varied between different Sox10 target gene promoters, indicating that Nmi function in vivo may be promoter-specific. PMID- 16214169 TI - Crystal structure of an archaeal peroxiredoxin from the aerobic hyperthermophilic crenarchaeon Aeropyrum pernix K1. AB - Peroxiredoxins (Prxs) are thiol-dependent peroxidases that catalyze the detoxification of various peroxide substrates such as H2O2, peroxinitrite, and hydroperoxides, and control some signal transduction in eukaryotic cells. Prxs are found in all cellular organisms and represent an enormous superfamily. Recent genome sequencing projects and biochemical studies have identified a novel subfamily, the archaeal Prxs. Their primary sequences are similar to those of the 1-Cys Prxs, which use only one cysteine residue in catalysis, while their catalytic properties resemble those of the typical 2-Cys Prxs, which utilize two cysteine residues from adjacent monomers within a dimer in catalysis. We present here the X-ray crystal structure of an archaeal Prx from the aerobic hyperthermophilic crenarchaeon, Aeropyrum pernix K1, determined at 2.3 A resolution (Rwork of 17.8% and Rfree of 23.0%). The overall subunit arrangement of the A.pernix archaeal Prx is a toroid-shaped pentamer of homodimers, or an (alpha2)5 decamer, as observed in the previously reported crystal structures of decameric Prxs. The basic folding topology and the peroxidatic active site structure are essentially the same as those of the 1-Cys Prx, hORF6, except that the C-terminal extension of the A.pernix archaeal Prx forms a unique helix with its flanking loops. The thiol group of the peroxidatic cysteine C50 is overoxidized to sulfonic acid. Notably, the resolving cysteine C213 forms the intra-monomer disulfide bond with the third cysteine, C207, which should be a unique structural characteristic in the many archaeal Prxs that retain two conserved cysteine residues in the C-terminal region. The conformational flexibility near the intra-monomer disulfide linkage might be necessary for the dramatic structural rearrangements that occur in the catalytic cycle. PMID- 16214170 TI - Plasminogen activator inhibitor-2 is highly tolerant to P8 residue substitution- implications for serpin mechanistic model and prediction of nsSNP activities. AB - The serine protease inhibitor (serpin) superfamily is involved in a wide range of cellular processes including fibrinolysis, angiogenesis, apoptosis, inflammation, metastasis and viral pathogenesis. Here, we investigate the unique mousetrap inhibition mechanism of serpins through saturation mutagenesis of the P8 residue for a typical family member, plasminogen activator inhibitor-2 (PAI-2). A number of studies have proposed an important role for the P8 residue in the efficient insertion and stabilisation of the cleaved reactive centre loop (RCL), which is a key event in the serpin inhibitory mechanism. The importance of this residue for inhibition of the PAI-2 protease target urinary plasminogen activator (urokinase, uPA) is confirmed, although a high degree of tolerance to P8 substitution is observed. Out of 19 possible PAI-2 P8 mutants, 16 display inhibitory activities within an order of magnitude of the wild-type P8 Thr species. Crystal structures of complexes between PAI-2 and RCL-mimicking peptides with P8 Met or Asp mutations are determined, and structural comparison with the wild-type complex substantiates the ability of the S8 pocket to accommodate disparate side-chains. These data indicate that the identity of the P8 residue is not a determinant of efficient RCL insertion, and provide further evidence for functional plasticity of key residues within enzyme structures. Poor correlation of observed PAI-2 P8 mutant activities with a range of physicochemical, evolutionary and thermodynamic predictive indices highlights the practical limitations of existing approaches to predicting the molecular phenotype of protein variants. PMID- 16214171 TI - Assessment of HPA-axis function in posttraumatic stress disorder: pharmacological and non-pharmacological challenge tests, a review. AB - Posttraumatic stress disorder (PTSD) is typically accompanied by acute and chronic alterations in the stress response. These alterations have mostly been described in individuals under baseline conditions, but several studies have also used a challenge model to further assess the role of the hypothalamic-pituitary adrenal (HPA) axis in the stress response. This paper reviews common methodology and research findings on HPA function in PTSD, and discusses the pathophysiological mechanisms underlying these findings. We reviewed the literature and selected all English-language, human subject, data driven, pharmacological and non-pharmacological challenge studies pertaining to the HPA axis, and in vitro leukocyte glucocorticoid receptor studies in adult PTSD subjects. Studies using a non-pharmacological stress paradigm (cognitive stress, trauma reminders) to stimulate the HPA axis showed an exaggerated cortisol response in PTSD. The most widely used pharmacological challenge with consistent results was the low dose dexamethasone-suppression test (DST). These DST studies showed enhanced cortisol suppression in subjects with PTSD. Different hypotheses have been purported to explain the alterations in HPA axis functioning in PTSD. The results of the reviewed challenge tests, however, did not exclusively support one of the hypothesized mechanisms. Further research assessing hormones at all levels of the HPA axis at both baseline and at challenge conditions with a proper stratification of study population, will be necessary for a better understanding of stress-responsivity on the level of the HPA axis in PTSD. PMID- 16214172 TI - A review of neuroimaging studies in PTSD: heterogeneity of response to symptom provocation. AB - Different experiential, psychophysiological, and neurobiological responses to traumatic symptom provocation in posttraumatic stress disorder (PTSD) have been reported in the literature. Two subtypes of trauma response have been hypothesized, one characterized predominantly by hyperarousal and the other primarily dissociative, each one representing unique pathways to chronic stress related psychopathology. Recent PTSD neuroimaging findings in our own laboratory support this notion and are consistent with the view that grouping all PTSD subjects, regardless of their different symptom patterns, in the same diagnostic category may interfere with our understanding of posttrauma psychopathology. This review will integrate findings of different experiential, psychophysiological, and neurobiological responses to traumatic symptom provocation with the clinical symptomatology and the neurobiology of PTSD. PMID- 16214173 TI - Connecticut statewide sTEP wave evaluation. AB - INTRODUCTION: Selective Traffic Enforcement Programs (sTEPs) are a proven method to change motorists' behavior. Since 1997, the Connecticut DOT's Division of Highway Safety has organized a statewide seat belt enforcement program, with sTEP waves every three or four months. To date, 28 waves have been implemented. METHOD: Pre-wave and post-wave seat belt observation surveys are conducted by both state and municipal police across the state. Survey results, as well as a summary of all enforcement activity during the wave, are submitted for evaluation. RESULTS: Connecticut seat belt use has continued to rise from one wave to the next in a predictable "saw blade" pattern. CONCLUSIONS: The data clearly demonstrate that agencies that have participated in a greater number of waves have experienced the greatest increase in belt use. Belt use has not yet plateaued and additional sTEP enforcement seems indicated. However, evidence from other states suggests that a plateau may occur somewhere in the mid 80% range. IMPACT ON INDUSTRY: Should this occur, Connecticut will work toward strengthening the round the clock model, emphasizing the importance of aggressive primary enforcement. PMID- 16214174 TI - Exploiting genotypic differences to identify genes important for EAE development. AB - Experimental autoimmune encephalomyelitis (EAE) is an animal model of the human autoimmune disease multiple sclerosis (MS) and is primarily driven by T helper type 1 (Th1) cells. Interleukin (IL)-12 and interferon (IFN)-gamma are important cytokines involved in the differentiation and amplification of Th1 cells, however mice deficient in either IFN-gamma or IL-12 still develop EAE. We have used microarray analysis of EAE-affected CNS tissues in wild-type, IFN-gamma -/- and IL-12 -/- animals to identify genes critical for development of EAE. Over 500 genes were regulated in at least one genotype and over 94 genes were regulated in all three. Of those, 17 were also upregulated in spleen during the disease. We show that a majority of the genes regulated in EAE are also regulated in diseased regions of human MS tissues. The genes in the pool of 94 are more likely to be found regulated in MS patients than the genes regulated in only one or two of the mouse strains suggesting that analyzing gene expression under these multiple genetic conditions may lead to better identification of the genes critical for disease development. PMID- 16214175 TI - Mathematical models of the fate of lymphoma B cells after antigen receptor ligation with specific antibodies. AB - We formulate models of the mechanism(s) by which B cell lymphoma cells stimulated with an antibody specific to the B cell receptor (IgM) become quiescent or apoptotic. In particular, we aim to reproduce experimental results by Marches et al. according to which the fate of the targeted cells (Daudi) depends on the levels of expression of p21(Waf1) (p21) cell-cycle inhibitor. A simple model is formulated in which the basic ingredients are p21 and caspase activity, and their mutual inhibition. We show that this model does not reproduce the experimental results and that further refinement is needed. A second model successfully reproduces the experimental observations, for a given set of parameter values, indicating a critical role for Myc in the fate decision process. We use bifurcation analysis and objective sensitivity analysis to assess the robustness of our results. Importantly, this analysis yields experimentally testable predictions on the role of Myc, which could have therapeutic implications. PMID- 16214176 TI - Impaired plasma nitric oxide availability and extracellular superoxide dismutase activity in healthy humans with advancing age. AB - This study is aimed to verify the modifications of extracellular superoxide dismutase (EC-SOD) activity and its potential involvement on the mechanism responsible for the impairment of plasma nitric oxide (NO) availability occurring with advancing age in healthy humans. For this purpose, plasma samples were drawn from 40 healthy men, aged 20-92 years, in fasting state and used for measurements of stable end-product nitrite/nitrate (NOx), as expression of NO availability, EC SOD activity, thiobarbituric acid reactive substances (TBARS) as marker of lipid peroxidation, Trolox equivalent antioxidant capacity (TEAC) as a measure of plasma total antioxidant capacity, and in vitro susceptibility of low density lipoprotein (LDL) to copper-mediated oxidation, evaluated as lag time. As indicated by our results, advancing age was significantly related to decreased plasma values of NOx (r = -0.877, P < 0.001), EC-SOD activity (r = -0.888, P < 0.001), TEAC (r = -0.647, P < 0.001) and lag time (r = -0.621, P < 0.001) as well as to an increased plasma amount of TBARS (r = 0.858, P < 0.001). NOx plasma level resulted independently predicted by EC-SOD activity and age. EC-SOD activity, in turn, was determined by age and TEAC. Taken together, findings of the present study give further insight into the mechanism related to age associated endothelial dysfunction, indicating that the decreased EC-SOD activity may be involved in the progressive reduction of plasma NO availability with advancing age through the age-related impairment of oxidant/antioxidant balance. PMID- 16214177 TI - Effects of Pfaffia paniculata (Brazilian ginseng) extract on macrophage activity. AB - The roots of Pfaffia paniculata (Brazilian ginseng) have been indicated for the treatment of several diseases and as an analgesic and antiinflamatory drug. Treatment of mice with 200 mg/kg of the powdered root of P. paniculata reduced the Ehrlich ascitic volume [Matsuzaki, P., Akisue, G., Salgado Oloris, S.C., Gorniak, S.L., Zaidan Dagli, M.L., 2003. Effect of Pffafia paniculata (Brazilian ginseng) on the Ehrlich tumor on its ascitic form. Life Sciences, Dec 19; 74 (5), 573-579.]. One of the putative means to control the Ehrlich tumor growth is by increasing macrophage activity [Kleeb, S.R., Xavier, J.G., Frussa-Filho, R., Dagli, M.L.Z., 1997. Effect of haloperidol on the development of the solid Ehrlich tumor in mice. Life Sciences, 60 (4/5), 69-742.]. The aim of this study was to investigate experimentally the effects of the methanolic extract of P. paniculata roots on macrophage activity. Male mice received, by gavage, once a day, different doses (100, 250, or 500 mg/kg) of the methanolic extract of P. paniculata or filtered water, as control, for 10 days. Macrophage activity was evaluated through the phagocytosis index (PI), spreading index (SI), production of peroxide oxigen and nitric oxide. The peritoneal cells were activated with ip inoculation of Ehrlich ascitic cells, 24 h before the macrophage harvesting. The methanolic extract raised significantly the SI of mice from group of 500 mg/kg in comparison with the control group and group of 100 mg/kg. This raise of SI possibly induced the higher phagocytic activity observed in the experimental situation. Increased macrophage activity may be one of the effects contributing to inhibition of the Ehrlich ascitic tumor growth in mice. PMID- 16214178 TI - No change in cortical muscarinic M2, M3 receptors or [35S]GTPgammaS binding in schizophrenia. AB - Muscarinic M1, but not M4, receptors have been shown to be decreased in Brodmann's area (BA) 9 obtained postmortem from subjects with schizophrenia. This study extends that data by measuring levels of muscarinic M2 and M3 receptor protein and mRNAs in BA 9 and BA 40 from the same cohorts of subjects used in the study of M1 and M4 receptors. In addition, the ability of carbachol to stimulate muscarinic receptors that signal through the Gi/o G-proteins was measured in BA 9 from the same cohorts of subjects. There were no changes in levels of muscarinic M2 or M3 protein or M3 mRNA with diagnosis in either CNS region. M2 receptor mRNA could not be detected in BA 9 or BA 40. Finally, carbachol-stimulated GTPgammaS binding did not differ between the diagnostic cohorts in BA 9 (p = 0.64). These data add considerable weight to the argument that the muscarinic M1 receptor is the muscarinic receptor predominantly affected in BA 9 by the pathology of schizophrenia. Given the widespread changes in muscarinic receptors identified in the CNS of subjects of schizophrenia using functional neuroimaging it remains possible that receptors other than the M1 receptor may be altered in different CNS regions. PMID- 16214179 TI - Chronic administration of ethyl docosahexaenoate decreases mortality and cerebral edema in ischemic gerbils. AB - Dietary docosahexaenoic acid (DHA) intake can decrease the level of membrane arachidonic acid (AA), which is liberated during cerebral ischemia and implicated in the pathogenesis of brain damage. Therefore, in the present study, we investigated the effects of chronic ethyl docosahexaenoate (E-DHA) administration on mortality and cerebral edema induced by transient forebrain ischemia in gerbils. Male Mongolian gerbils were orally pretreated with either E-DHA (100, 150 mg/kg) or vehicle, once a day, for 4 weeks and were subjected to transient forebrain ischemia by bilateral common carotid occlusion for 30 min. The content of brain lipid AA at the termination of treatment, the survival ratio, change of regional cerebral blood flow (rCBF), brain free AA level, thromboxane B(2) (TXB(2)) production and cerebral edema formation following ischemia and reperfusion were evaluated. E-DHA (150 mg/kg) pretreatment significantly increased survival ratio, prevented post-ischemic hypoperfusion and attenuated cerebral edema after reperfusion compared with vehicle, which was well associated with the reduced levels of AA and TXB(2) in the E-DHA treated brain. These data suggest that the effects of E-DHA pretreatment on ischemic mortality and cerebral edema could be due to reduction of free AA liberation and accumulation, and its metabolite synthesis after ischemia and reperfusion by decreasing the content of membrane AA. PMID- 16214180 TI - Chronic treatment of DA-8159, a new phosphodiesterase type V inhibitor, attenuates endothelial dysfunction in stroke-prone spontaneously hypertensive rat. AB - This study examined the effects of chronic treatment of a new phosphodiesterase type 5 inhibitor, DA-8159, on endothelial dysfunction in stroke-prone spontaneously hypertensive rats (SHR-SP). Six-week-old male SHR-SP were divided into 4 groups; vehicle control, DA-8159 1, 3, and 10 mg/kg/day. During a 32-week experimental period, the animals were administered DA-8159 orally and fed a 4% NaCl-loaded diet. The systolic blood pressure was measured every two weeks throughout the experimental period using the tail-cuff method. At the end of experiments, the vascular function (acetylcholine-induced vasodilation) in the endothelium-intact aortic rings was investigated. In addition, the mortality, the left ventricular hypertrophy index, the plasma parameters and the incidence of a cerebral infarction were assessed. In the DA-8159 treated-rats, the vascular reactivity improved significantly in a dose-dependent manner. Although DA-8159 did not alter the elevation of the systolic blood pressure directly, the 3 and 10 mg/kg/day DA-8159 treatment delayed the early death caused by stroke. DA-8159 significantly reduced the left ventricular heart weight/body weight ratio compared with the vehicle control group. Furthermore, the DA-8159 treatment significantly increased the plasma nitric oxide, cGMP, and the total antioxidative status. The DA-8159 treatment also reduced the occurrence of stroke associated cerebral damage. These results indicate that DA-8159 can ameliorate an endothelial dysfunction-related vascular injury. Therefore, pharmacological intervention aimed at attenuating an endothelial dysfunction is important and might be useful in both preventing and treating endothelial dysfunction-related complications. PMID- 16214181 TI - Mad2beta, an alternative variant of Mad2 reducing mitotic arrest and apoptosis induced by adriamycin in gastric cancer cells. AB - Mad2beta is an alternative splicing variant of spindle checkpoint gene mad2, which was previously found by us and was related to the drug resistance in gastric cancer cells. In this paper, we explored the molecular mechanisms that Mad2beta variant promoted the formation of multidrug resistance in gastric cancer cells. We found that Mad2beta variant was detected only in the two human drug resistant gastric cancer cell sublines SGC7901/VCR and SGC7901/ADR, and it did not appear in its parental cell line SGC7901 and other detected gastric cancer cell lines. Expressions of Mad2 mRNA and protein in SGC7901 cells transfected with Mad2beta, SGC7901/VCR and SGC7901/ADR were significantly lower than that in SGC7901 cells. Moreover, SGC7901 cells overexpressing Mad2beta variant became more resistant to adriamycin, vincristine and mitomycin by abrogating mitotic arrest and apoptosis. This suggests that expression of Mad2beta variant decreases the relative expression of efficient MAD2, which may help gastric cancer cells to develop the phenotype of multidrug resistance. PMID- 16214182 TI - Persistence of viral infections on the population level explained by an immunoepidemiological model. AB - We consider a mathematical model of viral spread in a population based on an immune response model embedded in an epidemic network model. The immune response model includes virus load and effector and memory T cells with two possible outcomes depending on parameters: (a) virus clearance and establishment of immune memory and (b) establishment of a non-zero viral presence characterized with increased T-cell concentrations. Isolated individuals can have different immune system parameters and, after a primary infection, can either return to the infection-free state or develop persistent or chronic infection. When individuals are connected in the network, they can reinfect each other. We show that the virus can persist in the epidemic network for indefinite time even if the whole population consists of individuals that are able to clear the virus when isolated from the network. In this case a few individuals with a relatively weak immune response can maintain the infection in the whole population. These results are in contrast to implications of classical epidemiological models that a viral epidemic will end if there is no influx of new susceptibles and if individuals can become immune after infection. PMID- 16214183 TI - Ito versus Stratonovich calculus in random population growth. AB - The context is the general stochastic differential equation (SDE) model dN/dt=N(g(N)+sigmaepsilon(t)) for population growth in a randomly fluctuating environment. Here, N=N(t) is the population size at time t, g(N) is the 'average' per capita growth rate (we work with a general almost arbitrary function g), and sigmaepsilon(t) is the effect of environmental fluctuations (sigma>0, epsilon(t) standard white noise). There are two main stochastic calculus used to interpret the SDE, Ito calculus and Stratonovich calculus. They yield different solutions and even qualitatively different predictions (on extinction, for example). So, there is a controversy on which calculus one should use. We will resolve the controversy and show that the real issue is merely semantic. It is due to the informal interpretation of g(x) as being an (unspecified) 'average' per capita growth rate (when population size is x). The implicit assumption usually made in the literature is that the 'average' growth rate is the same for both calculi, when indeed this rate should be defined in terms of the observed process. We prove that, when using Ito calculus, g(N) is indeed the arithmetic average growth rate R(a)(x) and, when using Stratonovich calculus, g(N) is indeed the geometric average growth rate R(g)(x). Writing the solutions of the SDE in terms of a well defined average, R(a)(x) or R(g)(x), instead of an undefined 'average' g(x), we prove that the two calculi yield exactly the same solution. The apparent difference was due to the semantic confusion of taking the informal term 'average growth rate' as meaning the same average. PMID- 16214184 TI - Learning by doing versus learning by thinking: An fMRI study of motor and mental training. AB - Previous studies have documented that motor training improves performance on motor skill tasks and related this to altered functional brain activity in cerebellum, striatum, and frontal motor cortical areas. Mental training can also improve the performance on motor tasks, but the neural basis of such facilitation is unclear. The purpose of the present study was to identify neural correlates of training-related changes on a finger-tapping task. Subjects were scanned twice, 1 week apart, with fMRI while they performed two finger-tapping sequences with the left hand. In-between scans, they practiced daily on one of the sequences. Half of the participants received motor training and the other half received mental training (motor imagery). Both training procedures led to significant increases in tapping performance. This was seen for both the trained and the untrained sequence (non-specific effect), although the gain was larger for the trained sequence (sequence-specific effect). The non-specific training effect corresponded to a reduction in the number of activated areas from an extensive set of brain regions prior to training to mainly motor cortex and cerebellum after training. The sequence-specific training effect involved the supplementary motor area and the cerebellum for motor training and visual association cortex for mental training. We conclude that gains following motor and mental training are based on distinct neuroplastic changes in the brain. PMID- 16214185 TI - Centella asiatica treatment during postnatal period enhances learning and memory in mice. AB - Present investigation was planned to evaluate the nootropic effect of Centella asiatica. Three months old male Swiss albino mice were injected orally with graded doses (200, 500, 700, 1000 mg/kg body weight) of C. asiatica aqueous extract for 15 days to select an effective dose for nootropic studies. Animals were tested in radial arm maze to assess the learning and memory performance. Based on these results, mice were treated orally with 200 mg/kg of C. asiatica for 15 days from day 15 to day 30 post partum (p.p.) and the nootropic effect was evaluated on the 31st day and 6 months p.p. The behavioral (open field, dark/bright arena, hole board and radial arm maze tests), biochemical (acetylcholine esterase activity) and histological studies (dendritic arborization) were carried out. Performance of juvenile and young adult mice was significantly improved in radial arm maze and hole board tests, but locomotor activity did not show any change compared to control. Treatment resulted in increased acetylcholine esterase activity in the hippocampus. Dendritic arborization of hippocampal CA3 neurons was also increased in terms of intersections and branching points, both at one month and 6 months. Results of the present investigation show that treatment during postnatal developmental stage with C. asiatica extract can influence the neuronal morphology and promote the higher brain function of juvenile and young adult mice. PMID- 16214186 TI - Cardiovascular disease prevention in Russia: challenges and opportunities. AB - OBJECTIVES: This paper examines the challenges that have faced the implementation of cardiovascular disease (CVD) prevention programmes in Russia since the 1980s in two regions of Russia, and identifies opportunities for improving such efforts. STUDY DESIGN AND METHODS: This study used a mixed methods design consisting of archival data review of health-related policy documents and legislation, and key informant interviews. RESULTS: CVD is the leading cause of death in Russia, with rapid increases in prevalence following the collapse of the Soviet Union in 1991. The MONICA, Tacis and CINDI programmes have played major roles in the development of non-communicable and CVD prevention policies and programmes in Russia since the 1980s. These programmes have assisted in policy and guideline development, and programme implementation. However, significant barriers in realizing such policies and sustaining prevention programmes have been encountered. CONCLUSION: Numerous barriers exist in developing and implementing CVD prevention programmes in Russia. More government engagement backed by strong public support is necessary in order to sustain and build capacity for CVD prevention in Russia. PMID- 16214187 TI - Understanding the spatial diffusion process of severe acute respiratory syndrome in Beijing. AB - OBJECTIVES: To measure the spatial contagion of severe acute respiratory syndrome (SARS) in Beijing and to test the different epidemic factors of the spread of SARS in different periods. METHODS: A join-count spatial statistic study was conducted and the given hypothetical processes of the spread of SARS in Beijing were tested using various definitions of 'joins'. RESULTS: The spatial statistics showed that of the six diffusion processes, the highest negative autocorrelation occurred in the doctor-number model (M-5) and the lowest negative autocorrelation was found in the population-amount model (M-3). The results also showed that in the whole 29-day research period, about hour or more days experienced a significant degree of contagion. CONCLUSIONS: Spatial analysis is helpful in understanding the spatial diffusion process of an epidemic. The geographical relationships were important during the early phase of the SARS epidemic in Beijing. The statistic based on the number of doctors was significant and more informative than that of the number of hospitals. It reveals that doctors were important in the spread of SARS in Beijing, and hospitals were not as important as doctors in the contagion period. People are the key to the spread of SARS, but the population density was more significant than the population size, although they were both important throughout the whole period. PMID- 16214188 TI - Analysis of responses of Long Beach, California residents to the Smoke-free Bars Law. AB - OBJECTIVES: We investigated survey responses to the Smoke-free Bars Law by residents of Long Beach, California (population 460,000), a city that reflects the state's diverse population. The research specifically aimed to determine: (1) residents' approval for the 1998 California Smoke-free Bars Law when it was implemented; and (2) changes in approval between baseline and 2-year follow-up. Data were also assessed for the demographic characteristics of the respondents and whether the respondents were self-acknowledged smokers or non-smokers. STUDY DESIGN: A random telephone survey was conducted in 1998 and 2000 in Long Beach to determine the degree of community support for the 1998 state law that prohibited smoking in all workplaces including alcohol-serving establishments. The numbers analysed were 784 in 1998 and 1237 in 2000. METHODS: Statistical analyses used in this research included univariate frequency distributions and logistic regression for 1998 and 2000. RESULTS: The major findings were as follows. Overall community approval for the 1998 state law increased from 65.2% in 1998 to 72.6% in 2000. Over this period, the rate of approval by smokers increased from 20.6% to 37.1%, and the rate of approval by non-smokers increased from 74.5% to 80.3%. CONCLUSIONS: The general public in a large city in California strongly approve of the prohibition of smoking in all indoor public places. This strong endorsement has major public health implications. PMID- 16214189 TI - Prevalence and correlates of hepatitis C infection among users of North America's first medically supervised safer injection facility. AB - BACKGROUND: North America's first medically supervised safer injection facility (SIF) for illicit drug users was opened in Vancouver, Canada on 22 September 2003. We examined the prevalence and correlates of hepatitis C (HCV) infection among a representative cohort of SIF users. METHODS: Users of the Vancouver SIF were selected at random and asked to enrol in the Scientific Evaluation of Supervised Injecting (SEOSI) cohort. At baseline, venous blood samples were collected and an interviewer-administered questionnaire was performed. Participants who were HCV-positive were compared with HCV-negative subjects using bivariate and logistic regression analyses. RESULTS: Between 1 December 2003 and 30 July 2004, 691 participants were enrolled into the SEOSI cohort, among whom 605 (87.6%) were HCV-positive at baseline. Factors independently associated with HCV infection in logistic regression analyses included: involvement with the sex trade [adjusted odds ratio (AOR) 3.7, 95% confidence interval (CI) 2.1-6.1], history of borrowing syringes (AOR 1.8, 95%CI 1.1-2.9), and history of incarceration (AOR 2.6, 95%CI 1.5-4.4). Daily heroin use was protective against HCV infection (AOR 0.6, 95%CI 0.3-0.9). CONCLUSION: The SIF has attracted injection drug users with a high burden of HCV infection and a substantial proportion of uninfected individuals. Although cross-sectional, this study provides some insight into historical risks for HCV infection among this population, and prospective follow-up of this cohort will be useful to determine if use of the SIF is associated with reduced risk behaviour and HCV incidence. PMID- 16214190 TI - Evaluation of a rapid screening method for detection of antimicrobial resistance in the commensal microbiota of the gut. AB - The assessment of antimicrobial resistance among commensal bacteria is an indicator of the spread of antimicrobial resistance. Rapid screening methods for detection of antimicrobial-resistant faecal Escherichia coli directly on MacConkey plates have been successfully adopted but suffer from lack of standardisation. The purpose of this study was to evaluate a direct plating method (DPM) for detection of antimicrobial-resistant faecal E. coli and to compare it with a conventional method. Faecal samples were collected from 71 healthy children from Peru and Bolivia. In the DPM, a faecal swab was directly plated onto a MacConkey agar plate and antimicrobial disks were applied onto the seeded plate. Raw data were obtained by direct reading of the plate and were subjected to confirmatory analysis. Good concordance between the DPM and a conventional method was observed in detecting carriage of resistant E. coli, with a higher sensitivity for the DPM. Analysis of the results allowed interpretive criteria to be defined for DPM raw data. The DPM showed good sensitivity and specificity at very low cost (ten times cheaper than the conventional method) to investigate the faecal carriage of drug-resistant E. coli. It may represent a useful tool to conduct large-scale resistance surveillance studies and to monitor resistance control programmes cost effectively, particularly in low-resource countries. PMID- 16214191 TI - Using gender analysis to build voluntary counselling and testing responses in Kenya. AB - The rapid expansion of voluntary counselling and testing (VCT) for HIV in sub Saharan Africa has led to concerns over the quality and equity of the services. Kenya has seen an unprecedented scale-up of VCT, and valuable lessons have been learnt at national as well as at district and community levels. We combined quantitative and qualitative research methodology and showed how the results of gender analysis can be used to develop equity in VCT scale-up. A gender disaggregated analysis of VCT client data was conducted for the first 8 months of 2003. These quantitative data revealed that despite an increased vulnerability to HIV, women are underrepresented in VCT sites in all settings in Kenya. Our data also showed that women were also less likely to use condoms or to take home condoms after a VCT visit than their male counterparts. Further exploration through in-depth qualitative work with women and men allowed a better understanding of the reasons behind gender differences in Kenyan VCT sites and helped to develop strategies to address gender inequity. We conclude that there is an ongoing need to mainstream gender in monitoring and evaluation strategies to ensure services meet the needs and priorities of all groups. PMID- 16214192 TI - How can the community contribute in the fight against HIV/AIDS and tuberculosis? An example from a rural district in Malawi. AB - This paper describes (a) the experience of initiating community involvement in HIV/AIDS and tuberculosis (TB) activities in a rural district in Malawi and (b) some of the different ways in which the community is contributing in the fight against these two diseases and the outcomes of their involvement. During a 2-year period, a total of 21,358 (41%) of 52,510 HIV tests performed at voluntary counselling and HIV testing (VCT) sites in the district were conducted by lay community counsellors. A team of 465 community volunteers, 1,362 trained family caregivers and 9 community nurses provided care and support to 5,106 HIV-positive individuals, of whom 2,006 (39%) were in WHO stage III or IV. All those in WHO stage III or IV were on co-trimoxazole prophylaxis and 895 (45%) of these were also on antiretroviral treatment. A total of 2,714 TB patients, of whom 1627 (60%) were HIV-positive, also received care and support. A total of 1,694 orphans were trained in vocational skills. Twelve vegetable gardens and three maize farms were set up, and pre-school activities were organised for 900 orphans. Communities can play an important contributory role in reducing the burden of HIV/AIDS and TB and in mitigating its impact. Despite this, community resources in most settings are often under-exploited and their role remains undefined. PMID- 16214193 TI - Current issues in the prevention of mother-to-child transmission of HIV-1 infection. AB - Worldwide every year approximately 750,000 children become infected with HIV, mostly through mother-to-child transmission (MTCT). Without specific interventions, the rate of MTCT is approximately 15-20%, with prolonged breastfeeding doubling the rate to 35-40%. Current approaches to intervention to reduce the risk of MTCT focus mainly on antiretroviral prophylaxis during pregnancy, labour and in the early neonatal period, but in some settings also on delivery procedures and avoidance of breastfeeding. To reduce the risk of breastfeeding transmission, shortening the breastfeeding period and/or encouraging exclusive breastfeeding has been suggested. MTCT rates of less than 2% are now reported from countries where antiretroviral prophylaxis, elective Caesarean section and refraining from breastfeeding can be applied, whilst in settings where refraining from breastfeeding is not feasible or safe and where elective Caesarean section is also not a safe option, peripartum antiretroviral therapy can halve the risk to levels of approximately 10% at 6 weeks, although further acquisition of infection through breastfeeding substantially increases the overall rate to 20% or more. In light of the high mortality in all children of HIV-infected women, programmes for prevention of MTCT should be monitored not only in terms of HIV transmission avoided but also in terms of child survival. PMID- 16214194 TI - Looking within the household: gender roles and responses to malaria in Ghana. AB - Studies of factors affecting treatment-seeking behaviour for malaria have rarely considered the influence of gender roles and relations within the household. This study supported district-level government workers in the Volta Region of Ghana in conducting a situational analysis of gender inequities in relation to the malaria burden and access to healthcare services for malaria in one community in their district. Qualitative and participatory methods, such as focus group discussions, in-depth individual interviews and ranking exercises, were used. The study found that women who lacked either short- or long-term economic support from male relatives, or disagreed with their husbands or family elders about appropriate treatment-seeking, faced difficulties in accessing health care for children with malaria. This illustrates the significant influence of women's access to resources and decision-making power on treatment-seeking behaviour for children with febrile illnesses, and the importance of approaching malaria management in the community or household from a gender perspective. PMID- 16214195 TI - Pleomorphism of the clinical manifestations of neurocysticercosis. AB - Neurocysticercosis (NCC) remains a major public health problem in developing countries as it is the most common helminthic infection of the central nervous system. Clinical manifestations are non-specific and pleomorphic. Case reports on uncommon presentations of NCC are few. We report six interesting cases of NCC with unusual clinical presentation that demonstrate this spectrum of pleomorphism. These include extrapyramidal disease (parkinsonism and focal dystonia), Kluver-Bucy syndrome, Weber's syndrome, dementia and cortical blindness. The clinical details and possible mechanisms for the uncommon presentations are also discussed. Thus, a high level of suspicion should be kept for NCC, especially in endemic zones and developing countries. PMID- 16214196 TI - Repeated treatment of cystic echinococcosis in patients with a long-term immunological response after successful surgical cyst removal. AB - Six cystic echinococcosis patients underwent surgery for the removal of echinococcal cysts. All were treated with albendazole prior to and following treatment. After surgery, no cysts were detected in five of the six patients examined. Both ELISA and immunoblot analysis have been used to determine specific IgG, IgG4 and IgE activities. Total elimination of IgG and IgG4 was not achieved in any of the patients studied. Prior to the first surgery/treatment, specific IgG, IgG4 and IgE antibodies were demonstrated in all patients, except one who did not show any IgE activity. The first treatment was followed by highly elevated IgE in two patients; in one of them it was further combined with an apparent decrease in IgG activity. Repeated treatment with albendazole given 0.8 8.5 years after the first treatment/surgery was followed by either moderate or highly reduced IgE activity in two patients, respectively, and a slight increase in IgG4 in another patient. A third course of treatment, given 2-2.5 years after the second treatment, barely affected the antibody activities. The present study suggests that anti-echinoccocal antibody activity may remain high many years after successful cyst removal. Determination of IgG, IgG4 and IgE responses is preferable for the assessment of treatment results. The presence of anti echinococcal antibodies after surgery with no cyst detection does not necessarily indicate an active echinococcal infection. PMID- 16214197 TI - Inhibition of dengue virus translation and RNA synthesis by a morpholino oligomer targeted to the top of the terminal 3' stem-loop structure. AB - Dengue virus (DEN) is a major public health problem worldwide and causes a spectrum of diseases, for which no antiviral treatments exist. Peptide-conjugated phosphorodiamidate morpholino oligomers (P-PMOs) complementary to the DEN 5' stem loop (5'SL) and to the DEN 3' cyclization sequence (3'CS) inhibit DEN replication, presumably by blocking critical RNA-RNA or RNA-protein interactions involved in viral translation and/or RNA synthesis. Here, a third P-PMO, complementary to the top of the 3' stem-loop (3'SLT), inhibited DEN replication in BHK cells. Using a novel DEN2 reporter replicon and a DEN2 reporter mRNA, we determined that the 5'SL P-PMO inhibited viral translation, the 3'CS P-PMO blocked viral RNA synthesis but not viral translation, and the 3'SLT P-PMO inhibited both viral translation and RNA synthesis. These results show that the 3'CS and the 3'SL domains regulate DEN translation and RNA synthesis and further demonstrate that P-PMOs are potentially useful as antiviral agents. PMID- 16214198 TI - Coherent motion detection in preschool children at family risk for dyslexia. AB - We tested sensitivity to coherent motion (CM) in random dot kinematograms in a group of 5-year-old preschool children genetically at risk for dyslexia, compared to a group of well-matched control children. No significant differences were observed, either in a group analysis or in an individual deviance analysis. Nonetheless, CM-thresholds were significantly related to emerging orthographic skills. In a previous study on the same subjects (Boets, Wouters, van Wieringen, & Ghesquiere, in press), we demonstrated that both risk groups already differed on measures of phonological awareness and letter knowledge. Moreover, auditory spectral processing (especially 2 Hz FM detection) was significantly related to phonological ability. In sum, the actual visual and previous auditory data combined, seem to suggest an exclusive relation between CM sensitivity and orthographic skills on the one hand, and FM sensitivity and phonological skills on the other. PMID- 16214199 TI - The role of attention in binding shape to color. AB - Pictures of easily-identifiable objects with novel colors (e.g. a blue frog) or of forms with arbitrary colors (e.g. a green triangle) were presented briefly at 10.6 degrees eccentricity. Stimuli had strong outlines and vivid fill colors (red, green, yellow, blue, or purple). The same pictures were repeated once in each block of 30 trials for 6, 9, or 12 blocks, and recognition was probed after each block. Shapes were acquired quickly, within 3-4 blocks, whether attention was focused on the pictures or split to a demanding foveal task. Color-shape acquisition was also fast with focused attention, but stabilized at a low level with split attention. Delaying the foveal task restored color-shape acquisition. We suggest that attention facilitates the creation and maintenance of novel color shape bindings in the visual periphery; without attention, binding is less effective. PMID- 16214200 TI - Development and application of functional assays for freshwater dissolved organic matter. AB - A series of 11 standardised, reproducible, assays have been developed of physico chemical functions of dissolved organic matter (DOM) in freshwaters. The assays provide quantitative information on light absorption, fluorescence, photochemical fading, pH buffering, copper binding, benzo(a)pyrene binding, hydrophilicity and adsorption to alumina. To obtain DOM for the assays, a 45 L sample of filtered freshwater was rotary-evaporated to reduce the volume to ca. 500 cm3. The concentrate was then passed through a strong cation exchanger, in the Na+ form, to remove alkaline-earth cations, and then through 0.7 and 0.2 microm filters. Eight samples, two each from a lake and three streamwaters, were processed. The yields of dissolved organic carbon (DOC) ranged from 70% to 107% (average 91%). The samples of DOM, stored in the dark at 4 degrees C, retained their functional assay characteristics for up to 7 months. When assaying the concentrates, parallel assays were performed with Suwannee River fulvic acid (SRFA), as a quality control standard. For most of the assays, the results for eight freshwater DOM samples are similar to those obtained with SRFA, the chief exception being the greater hydrophilicity of the DOM samples. For eight of the assays, variability among the DOM samples is significantly (p < 0.01) greater than can be explained by analytical error, i.e. by comparison with results for the SRFA quality standard; the three exceptional assays are photochemical fading, copper binding and benzo(a)pyrene binding. The two lakewater samples studied gave the most extreme assay results, probably because of the influence of phytoplankton-derived DOM. Significant correlations of hydrophilicity and adsorption with optical absorbance may mean that some DOM functional properties can be predicted from comparatively simple measurements. PMID- 16214201 TI - Prediction of environmental partition coefficients and the Henry's law constants for 135 congeners of chlorodibenzothiophene. AB - Polychlorinated dibenzothiophenes (PCDTs) could be classified as persistent organic pollutants (POPs) in the environment and are particularly interesting due to their structural resemblance to highly toxic dioxins. We show here some basic environmental properties such as n-octanol water (K(OW)), n-octanol/air (K(OA)) and air/water (K(AW)) partition coefficients as well as Henry's law constants (K(H)) for all 135 congeners of chlorodibenzothiophene. Predictions were made by regression of principal components (PCR) and with aid of a set of standard chemicals, for which physical-chemical properties are well featured. Computed K(OW), K(OA), K(AW) and K(H) values for mono-CDTs ranged, respectively, between 4.66 and 4.71, 7.48 and 7.55, -2.84 and -2.82, 3.56 and 3.74; for di-CDTs between 5.02 and 5.28, 8.03 and 8.29, -3.01 and -2.95, 2.42 and 2.75; for tri-CDTs between 5.53 and 5.70, 8.65 and 8.87, -3.2 and -3.11, 1.58 and 1.92; for tetra CDTs between 5.95 and 6.13, 9.27 and 9.50, -3.39 and -3.27, 1.02 and 1.33; for penta-CDTs between 6.38 and 6.51, 9.88 and 10.05, -3.54 and -3.45, 0.72 and 0.88; for hexa-CDTs between 6.83 and 6.97, 10.54 and 10.66, -3.72 and -3.64, 0.47 and 0.56; for hepta-CDTs between 7.28 and 7.35, 11.12 and 11.20, -3.81 and -3.87, 0.33 and 0.38; for octa-CDT 7.74, 11.78, -4.04 and 0.23. An estimated value of the three types of partition coefficient and Henry's law constants suggest that polychlorinated dibenzothiophenes are lipophilic and semi-volatile persistent organic pollutants. Their mobility in the environment seems to be very similar to that of some well-known POPs such as polychlorinated dibenzofurans, -dibenzo-p dioxins, and -biphenyls or organochlorine pesticides. PMID- 16214202 TI - River flow and associated transport of sediments and solutes through a highly urbanised catchment, Bradford, West Yorkshire. AB - The hydrological characteristics of catchments become drastically modified in response to urbanisation. The total contributions and dynamics of runoff, suspended sediment and solutes may change significantly and have important implications downstream where they may affect flooding, instream ecological habitat, water quality and siltation of river channels and lakes. Although an appreciation of the likely hydrological changes is crucial for effective catchment management they are still poorly understood. In this paper we present data from a network of river monitoring stations throughout the heavily urbanised Bradford catchment, West Yorkshire. Sites are upstream, within and downstream of the highly urbanised central part of the catchment. Flow, turbidity (calibrated to suspended sediment concentration) and specific conductance (surrogate for solute concentration), logged at 15-min intervals, are presented for a 12-month period (June 2000 to June 2001). The total amounts and dynamics of flow, solute and suspended sediment transport were investigated. Estimated total flow and suspended sediment transport for the monitoring period were found to be high in response to the high total rainfall. Flow and sediment transport regimes were extremely 'flashy' throughout the catchment and became increasingly flashy in a downstream direction. Suspended sediment discharged from the Bradford subcatchment makes an important contribution to downstream sediment transport on the river Aire at Beal. Data suggest that the urbanised part of the Bradford catchment is extremely important in contributing solutes to the Beck (river). Although flow and sediment are also contributed to the Bradford Beck in the urbanised part of the catchment the data suggest that significant amounts may enter the combined sewer system and bypass the river. Understanding the spatial and temporal variations of flow and the transport of suspended sediment and solutes in rivers in urbanized subcatchments is crucial to their effective management and monitoring. Furthermore, this knowledge may be extremely important to the management and monitoring of downstream rivers in large scale mixed catchments. PMID- 16214203 TI - Is guidewire exchange a better approach for subclavian vein re-catheterization for chronic hemodialysis patients? AB - BACKGROUND: The objectives of this study were to compare outcomes and survival rates of subclavian vein re-catheterization through guide wire exchange (GWE) or de novo insertion (DN). MATERIALS AND METHODS: The study was conducted in a retrospective manner. Medical records of 36 patients who received percutaneous subclavian vein re-catheterization for hemodialysis in our institution during the period from April 1, 2001 to September 30, 2004 were reviewed. All patients had at least 2 catheter insertions records in our institute. Incidences of adverse events (infection, thrombosis) were compared between GWE and DN groups using x2 test. Predictors for adverse event occurrences were analyzed using logistic regression models. Cox proportional hazard model was used to investigate the predictors for adverse event-free catheter days. Kaplan-Meire survival curves were computed and compared using log rank test. RESULTS: Information were generated from 98 catheters (41 from DN, 57 from GWE groups). The average catheter usage was 2.8+/-0.9 devices per patient and the mean catheter-indwelling day was 125.4+/-129.5 days in this cohort. We found GWE group had significantly lower thrombosis rate (49.1% vs. 85.4% for DN group, P<0.000) in general. Surgical approach was a significant risk factors for catheter thrombosis (GWE vs. DN, odds ratio=0.261, P=0.05). The actuarial survival rates for GWE were > or =30 days, 85.4%; > or =60 days, 75.5%; > or =90 days, 64.5%; > or =180 days, 44.3%. The actuarial survival rates for DN were > or =30 days, 70.7%; > or =60 days, 58.5%; > or =90 days, 34.2%; > or =180 days, 18.4%. GWE group catheters had significantly higher catheter survival rates (P=0.0009). Mahukar catheter (hazard ratio 0.514, P=0.03), non-shock (hazard ratio 3.358, P=0.04), and older age (hazard ratio 0.958, P=0.026) were predictors of adverse event-free remaining catheter days. CONCLUSION: We suggest that GWE might be a favorable option over DN insertion when revised subclavian vein catheterization is inevitable. GWE can be performed repeatedly without compromising catheter outcomes. PMID- 16214204 TI - A polymorphism in the matrix metalloproteinase-1 gene promoter is associated with the prognosis of patients with ovarian cancer. AB - OBJECTIVE: The enzyme matrix metalloproteinase (MMP)-1 is involved in ovarian carcinogenesis. A common guanine insertion-deletion promoter polymorphism within the gene encoding MMP-1 (MMP1) has been suggested to be a candidate gene for ovarian cancer. We investigated whether this common polymorphism can also serve as independent prognostic parameter in a large series of affected women. METHODS: The MMP1 promoter polymorphism was examined in 151 Caucasian patients with epithelial ovarian cancer using polymerase chain reaction. Results were correlated with clinical data. RESULTS: No associations were ascertained between the MMP1 polymorphism and tumor stage (P = 1.0, odds ratio [OR] 1.08), lymph node involvement (P = 1.0, OR 0.8), tumor grading (P = 0.2, OR 0.5), and patient's age at diagnosis (P = 1.0, OR 1.04). Besides the clinically established prognosticators, tumor stage and histological grade, presence of the MMP1 polymorphism was associated with a shortened disease-free and overall survival in a univariate Kaplan-Meier analysis (P = 0.01) and a multivariate Cox regression model (P = 0.04). CONCLUSION: Presence of the MMP1 gene promoter polymorphisms was found to be a negative prognostic parameter in patients with ovarian cancer. PMID- 16214205 TI - Interactive dynamic graphical techniques for the exploration of functional data. AB - INTRODUCTION: The use of interactive dynamic graphics has become a common practice for the exploration of multidimensional data sets. The availability of powerful and inexpensive hardware and software for graphical computing makes the use of such techniques feasible for the examination of complex forms of data. This paper describes some simple techniques, which were implemented in the LISP STAT environment, for the visualization of functional data arising from studies of optical technologies used for the detection of cervical intraepithelial neoplasia or squamous intraepithelial lesions. MATERIALS AND METHODS: The methods demonstrated have been implemented in software coded in LISP-STAT, a free statistical computing package available for most computer systems. The data used in this paper are drawn from a previous study in which fluorescence spectroscopy was measured from cervical sites at 337 nm, 380 nm, and 460 nm excitation in cervical screening patients. The goal of the project is to explore biographical variables to better understand the biology of fluorescence. RESULTS: 199 measurements were taken in 55 women with normal Pap smears. The data are recorded as spectra showing the intensity of emission excitation versus emission in nanometers. Covariate variables available for analysis are current smoker vs. nonsmoker premenopausal vs. postmenopausal, tissue type (columnar, squamous, and transition zone), and age in years. Although the optical measurements show consistent changes between normal and abnormal tissue in individual patients, there is wide variation in the intensity of the measurements between patients, even for normal tissue. Patient age affects the fluorescent spectrum showing increasing intensity with increasing age. Menopausal status affects the fluorescent spectra coincidentally with age. Smoking and race do not appear to affect the spectra in this sample of patients. CONCLUSIONS: The use of interactive graphical techniques permits the data analyst to examine multidimensional data in intuitive ways. These explorations allow non statisticians to explore the data in a perceptive manner that may lead to new approaches in algorithm development for optical technologies. PMID- 16214206 TI - Rhabdomyosarcoma of the cervix in sisters with review of the literature. AB - OBJECTIVES: Pediatric embryonal heterologous rhabdomyosarcoma of the cervix is a rare tumor. METHODS: We present an interesting clinico-pathologic situation of two sisters presenting with pediatric embryonal heterologous rhabdomyosarcoma of the cervix. RESULTS: Pediatric embryonal heterologous rhabdomyosarcomas of the cervix are relatively uncommon. After a Pubmed search from 1952 to present, to our knowledge, this is the only report involving sisters presenting with this disease. Treatment has been extrapolated from collaborative groups such as Intergroup Rhabdomyosarcoma Study (IRS) Group so that optimal management may be achieved. CONCLUSIONS: We urge continued reporting of these rare tumors to enhance understanding if there may be a genetic component associated with them. PMID- 16214208 TI - Nasal administration of the calcium channel blocker diltiazem decreases food intake and attenuates weight gain in rats. AB - Food intake is normally influenced by a multitude of complex endogenous neurochemical systems, in addition to numerous external environmental stimuli, including olfaction. Since most olfactory neurons process odorant exposures through Ca2+-mediated mechanisms via Ca2+ channels, a novel approach at influencing the ingestive behaviors of animals might therefore involve altering olfactory acuity via Ca2+ channel blockade. We tested the ability of a Ca2+ channel blocker, diltiazem, to alter food intake in hyperphagic rats when administered using the intranasal (i.n.), intraperitoneal (i.p.), oral (p.o.) or intracerebroventricular (i.c.v.) routes of administration. Male Sprague Dawley rats, which had been food-deprived for 4 h at the beginning of the dark cycle, were administered different doses of diltiazem (0-8 mg/animal or 0-40 mg/kg) and the amounts of food consumed were measured. While food intake at 1, 2 and 4 h post drug administration was significantly decreased in a dose-dependent manner after i.n. administration, the i.p., p.o., and i.c.v. routes did not affect food intake. In another experiment, rats trained to eat their daily meal during the first 4 h at the onset of the dark cycle and treated daily with i.n. diltiazem (0 8 mg/animal) prior to food introduction exhibited a significantly decreased rate of weight gain in a dose-dependent manner over a 14-day period. Both i.n. and i.p. diltiazem significantly increased the plasma drug concentration at 1 h, however there was no significant difference between these routes of administration. Additional studies failed to demonstrate any detrimental effects of i.n. diltiazem (0-8 mg/animal) on conditioned taste aversion, locomotion or gross neurological/behavioral competence using the rota-rod test. While a local action on the nasal odorant receptors is most likely the site of diltiazem's action, further studies are needed to determine the exact mechanism of action of i.n. diltiazem. PMID- 16214207 TI - Immune responses to repetitive adenovirus-mediated gene transfer and restoration of gene expression by cyclophosphamide or etoposide. AB - BACKGROUND: One major concern about adenoviral vectors for repetitive gene delivery is the induction of an immune response to the vector, thus impeding effective gene transduction. METHODS: To assess the immune response to the adenoviral vector, repetitive gene dosing was performed into rhesus monkey cervix and C3H mouse skin using the adenoviral vector carrying the lacZ gene. Three repetitive intracervical injections of adenovirus-lacZ were done in the rhesus monkey at the intervals of 4 weeks. Gene expression on the second and third injection was completely suppressed. RESULTS: Anti-adenovirus IgG levels and neutralizing antibody titers in the rhesus monkey significantly increased after the first injection of adenovirus. In the C3H mouse, neutralizing antibody titers significantly increased after the first injection of adenovirus-lacZ at more than 10(8) plaque-forming unit (PFU). The repetitive expression of lacZ gene in the mouse skin markedly decreased when the second injection is done more than 2 weeks after the first injection. Chronic low-dose treatment with cyclophosphamide or etoposide markedly suppressed neutralizing antibody titers in the mouse serum and restored the gene expression in the mouse skin on the second and third injection. CONCLUSIONS: It is suggested that repetitive gene expression by adenovirus mediated transfer may be reduced by circulating neutralizing antibodies and could be restored by chronic low-dose treatment with cyclophosphamide or etoposide. PMID- 16214209 TI - Neuroprotective effects of Polygonum multiflorum on nigrostriatal dopaminergic degeneration induced by paraquat and maneb in mice. AB - The neuroprotective effects of Polygonum multiflorum extract (PME) and its two fractions, ethanol-soluble PME (PME-I) and -insoluble PME (PME-II), on the degeneration of nigrostriatal dopaminergic neurons induced by a combination of paraquat and maneb (PQMB) were investigated in male C57BL/6 mice. The mice were treated twice a week for 6 weeks with intraperitoneal injections of PQMB. This combination caused a reduction of spontaneous locomotor activity, motor incoordination, and declines of dopamine level in the striatum and tyrosine hydroxylase-positive neurons in the substantia nigra. Administration of PME and PME-I once daily for 47 days during 6 weeks of PQMB treatment and last 8 days after PQMB significantly attenuated the impairment of behavioral performance and the decrease in striatal dopamine level and substantia nigral tyrosine hydroxylase-positive neurons in the PQMB-treated animals, whereas the administration of PME-II had no effect on these behavioral, neurochemical and histological indices. The present findings suggest that PME has a beneficial influence on parkinsonism induced by PQMB and that the effects of PME are attributable to some substance(s) included in the ethanol-soluble fraction of PME (PME-I). PMID- 16214210 TI - Short- and long-term skin graft survival in cattle clones with different mitochondrial haplotypes. AB - In contrast to nuclear DNA, cytoplasmic genes may differ among cloned animals due to the presence of polymorphic mitochondrial DNA haplotypes in the host oocytes, raising doubts about histocompatibility among clones. Three bovine clones were generated by nuclear transfer; dermal fibroblasts from a fetus were used as donor cells, whereas oocytes from abbatoir-derived ovaries were used as recipient cells. The mitochondrial DNA (sequencing of coding and non-coding regions) and nuclear DNA (13 microsatellite markers) of cloned and control animals were characterized to identify potential polymorphisms. Skin auto- and allografts were transplanted on the adult clones and a non-related animal as a measure of immunological reactivity. Nuclear DNA of cloned animals was genetically identical but differed in all microsatellites of the non-related control. Amounts of donor cell mitochondrial DNA in the skin ranged from 1 to 2.6% among clones. Few differences in heteroplasmy were observed between skin and WBC of the clones, indicating limited mitochondrial DNA segregation in tissues during pre- and post natal development to adulthood. Sequencing of the remaining oocyte-derived mitochondrial DNA haplotype identified polymorphisms in coding and non-coding regions, confirming their origin from unrelated maternal lineages. Nonetheless, skin transplants between clones were accepted for the 92 d study period, whereas third-party grafts were rejected. In conclusion, the nuclear transfer-generated adult bovine clones used in this study were immunologically compatible with one another despite differences in their mitochondrial DNA haplotypes. PMID- 16214211 TI - Adolescents' perception of risk and challenge: a qualitative study. AB - Concern has been mounting about the increasing numbers of adolescents who engage in risky behaviours. This qualitative study aimed to identify adolescents' perception of risk and explored the factors that adolescents felt influenced their decisions about engaging in risky behaviours. The results indicated that adolescents perceived risk to be something where the outcome was uncontrollable. In contrast, challenges were thought of as having a known end point that was difficult to achieve. They considered themselves to be well-informed about health risks facing them, yet mentioned occasions when they lacked the ability to state that they did not want to engage in certain "risky" behaviours. The implications of the findings for future interventions are discussed. PMID- 16214212 TI - Structural analysis of the sheath of a sheathed bacterium, Leptothrix cholodnii. AB - Leptothrix cholodnii is an aerobic sheath-forming bacterium often found in oligotrophic and metal-rich aquatic environments. The sheath of this bacterium was isolated by selectively lysing the cells. Glycine and cysteine were the major amino acids of the sheath. The sheath was readily dissolved in hydrazine, and a polysaccharide substituted with cysteine was recovered from the solution. Galactosamine, glucosamine and galacturonic acid were detected in the hydrazinolysate by gas liquid chromatography analysis. FAB-MS analysis of the hydrazinolysate suggested a sugar sequence of HexN-GalA-HexN-HexN. Methylation linkage analysis revealed the presence of 4-linked GalA, 3-linked HexN and 4 linked HexN. The sulfhydryl groups of the sheath were used for labeling with the fluorogenic reagent, 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F). The labeled sheath (ABD-sheath) was partially hydrolyzed and three fluorescent fragments were purified by HPLC. One of them was identified as ABD-cysteine. The second one was found to be the ABD-cysteine tetramer. Another fragment was indicated to be a pentasaccharide substituted with ABD-cysteine by nuclear magnetic resonance (NMR) analysis. It can be assumed that the polysaccharide and peptide moieties of the sheath are connected by a cysteine residue. NMR analysis of the hydrazinolysate revealed that the polysaccharide moiety of the sheath was constructed from a pentasaccharide repeating unit containing 2-amino-2 deoxygalacturonic acid (GalNA), as shown below. -->4)-alpha-GalNA-(1-->4)-alpha-D GalN(p)-(1-->4)-alpha-D-GalA(p)-(1-->4)-beta-D-GlcN(p)-(1-->3)-beta-D-GalN(p)-(1- >. PMID- 16214213 TI - Factor analysis of water quality characteristics including trace metal speciation in the coastal environmental system of Chennai Ennore. AB - Statistical analysis of water quality parameters including trace metal speciation was undertaken with a view to seeing the interrelationship between different variables and also to identify probable source components in order to explain the pollution status of Chennai Ennore coastal environmental system. Factor analysis has been used in the present work. This is essentially a data reduction technique and will suggest how many variates are important to explain the observed variances in the data. The possible variances in the water quality parameters may be due to either sources of anthropogenic origin or natural variances due to the season or due to different biogeochemical processes that are taking place in the system. When this analysis was carried out with our data on water quality parameters in the above coastal environmental system, we found that the prominent factor or the first factor called the eutrophication factor explained 24.51% of the total variance (comprised of variables like DO, pH, SS, ammonia-N, phosphate and silicate). The second factor called the copper contamination factor explained 10.61% (comprised of variables like labile Cu, total Cu and particulate Cu) and the third factor called metal removal factor explained 10.11% (comprised of variables like particulate Zn, particulate Pb and water temperature) of the variances, respectively. Although there are four more factors, they are all having variances less than 10%. From this study, it is seen that eutrophication is the main source component of pollution to the surface waters of this estuary and its adjacent coastal waters compared to dissolved trace metals. PMID- 16214214 TI - Influence of air-sea fluxes on chlorine isotopic composition of ocean water: implications for constancy in delta37Cl--a statistical inference. AB - The behaviors of chlorine isotopes in relation to air-sea flux variables have been investigated through multivariate statistical analyses (MSA). The MSA technique provides an approach to reduce the data set and was applied to a set of 7 air-sea flux variables to supplement and describe the variation in chlorine isotopic compositions (delta37Cl) of ocean water. The variation in delta37Cl values of surface ocean water from 51 stations in 4 major world oceans--the Pacific, Atlantic, Indian and the Southern Ocean has been observed from -0.76 to +0.74 per thousand (av. 0.039+/-0.04 per thousand). The observed delta37Cl values show basic homogeneity and indicate that the air-sea fluxes act differently in different oceanic regions and help to maintain the balance between delta37Cl values of the world oceans. The study showed that it is possible to model the behavior of chlorine isotopes to the extent of 38-73% for different geographical regions. The models offered here are purely statistical in nature; however, the relationships uncovered by these models extend our understanding of the constancy in delta37Cl of ocean water in relation to air-sea flux variables. PMID- 16214215 TI - Mannose-binding lectin gene polymorphism and its impact on human immunodeficiency virus 1 infection. AB - Mannose-binding lectin (MBL) is a serum protein whose low concentration is associated with the occurrence of allele variants named MBL*B, MBL*C and MBL*D. The present study investigated the association between MBL gene polymorphism and the susceptibility to HIV-1 infection. The study of 145 HIV-1-infected subjects and 99 healthy controls showed the presence of alleles MBL*A, MBL*B and MBL*D, whose frequencies were 69%, 22% and 09% among patients and 71%, 13% and 16% among healthy controls, respectively. The presence of the variant MBL*B was associated with higher plasma viral load levels, suggesting the importance of the MBL gene polymorphism in the clinical evolution of HIV-1-infected patients. PMID- 16214216 TI - Effect of posterior juxtascleral triamcinolone acetonide on choroidal neovascular growth after photodynamic therapy with verteporfin. AB - OBJECTIVE: To assess if posterior juxtascleral application of 40 mg triamcinolone acetonide (TA), given at the same time as initial photodynamic therapy (PDT) for predominantly classic choroidal neovascularization (CNV) related to age-related macular degeneration affects lesion growth at 3 and 6 months. DESIGN: Comparative (nonrandomized) interventional study. PARTICIPANTS: The study group consists of 38 eyes of 38 patients. The control group consists of 73 eyes of 73 patients. METHODS: Comparison of 2 consecutive case series collected at different times. The study group had a posterior juxtascleral TA with their initial PDT treatment. The controls were treated with PDT alone. All patients were reviewed at 1, 3, and 6 months. MAIN OUTCOME MEASURES: Change in total lesion size; secondary outcomes: area of leak, best-corrected visual acuity, number of treatments, and intraocular pressure. RESULTS: There was significantly less growth of total lesion at 3 months (mean difference = 2.47 mm2; 95% confidence interval (CI): +1.22 to +3.72 mm2; P = 0.0002) and 6 months (mean difference = 2.88 mm2; 95% CI: +0.61 to +5.15 mm2; P = 0.0134) in patients given TA with PDT compared with PDT alone. There was also a significantly smaller residual area of leak at 3 months in the study group (mean difference = 1.07 mm2; 95% CI: +0.16 to +1.97 mm2; P = 0.02). At 6 months, the residual area of leak between the 2 groups became comparable (mean difference = 0.13 mm2; 95% CI = -1.59 to +1.33 mm2; P = 0.86). Mean number of letters lost on the logarithm of the minimum angle of resolution chart at 6 months was 9.1 letters (standard error of the mean [SEM] = 2.21) in the study group compared with 12.4 letters (SEM = 1.91) in the control group (P = 0.30). At 6 months, 10 of 36 eyes (27.8%) in the study group showed > or =15 letters loss, compared with 29 of 73 eyes (39.7%) in the control group. Intraocular pressure was raised in 4 of 38 eyes (10.5%). Fewer retreatments were required in the TA with PDT group (2.03 compared with 2.47 [P = 0.006]). CONCLUSIONS: Posterior juxtascleral placement of TA with PDT at baseline significantly reduces CNV growth at 3 and 6 months. Fewer retreatments were required. Visual outcome may be improved, although we did not show a statistically significant improvement with this sample size. A larger, randomized trial with longer follow-up is justified. PMID- 16214217 TI - Complexation of antimony (Sb(V)) with guanosine 5'-monophosphate and guanosine 5' diphospho-D-mannose: formation of both mono- and bis-adducts. AB - In spite of the extensive use of pentavalent antimony chemotherapy, the mechanism of its anti-leishmania action is still not clear. Here, we report the interactions of Sb(V), including the clinically used drug stibogluconate, with guanosine 5'-monophosphate (5'-GMP) and guanosine 5'-diphospho-d-mannose (5'-GDP mannose) in aqueous solution. The deprotonated hydroxyl groups (-OH) of the ribose ring are shown to be the binding site for Sb(V), probably via chelation. Both mono- and bis-adducts were formed as determined by NMR, high performance liquid chromatography (HPLC) and electrospray ionization mass spectrometry (ESI MS), and both of them are stable in the pH range of 4 to around 9.5. The formation of the mono-adduct (k(1)=1.67x10(-3) and 3.43x10(-3) mM(-1) min(-1) for Sb(5'-GMP) and Sb(5'-GDP-mannose), respectively, at 298 K) was 10-fold faster than that of the bis-adduct (k(2)=0.16x10(-3) and 0.21x10(-3) mM(-1) min(-1), for Sb(5'-GMP)(2) and Sb(5'-GDP-mannose)(2), respectively), and the mono-adduct was the major species in solution with the [bis-adduct]/[mono-adduct]<0.5. The reactions of stibogluconate with 5'-GMP and 5'-GDP-mannose were slower than that of antimonate under similar conditions. PMID- 16214218 TI - The significance of lipid composition for membrane activity: new concepts and ways of assessing function. AB - In the last decade or so, it has been realised that membranes do not just have a lipid-bilayer structure in which proteins are embedded or with which they associate. Structures are dynamic and contain areas of heterogeneity which are vital for their formation. In this review, we discuss some of the ways in which these dynamic and heterogeneous structures have implications during stress and in relation to certain human diseases. A particular stress is that of temperature which may instigate adaptation in poikilotherms or appropriate defensive responses during fever in mammals. Recent data emphasise the role of membranes in sensing temperature changes and in controlling a regulatory loop with chaperone proteins. This loop seems to need the existence of specific membrane microdomains and also includes association of chaperone (heat stress) proteins with the membrane. The role of microdomains is then discussed further in relation to various human pathologies such as cardiovascular disease, cancer and neurodegenerative diseases. The concept of modifying membrane lipids (lipid therapy) as a means for treating such pathologies is then introduced. Examples are given when such methods have been shown to have benefit. In order to study membrane microheterogeneity in detail and to elucidate possible molecular mechanisms that account for alteration in membrane function, new methods are needed. In the second part of the review, we discuss ultra-sensitive and ultra resolution imaging techniques. These include atomic force microscopy, single particle tracking, single particle tracing and various modern fluorescence methods. Finally, we deal with computing simulation of membrane systems. Such methods include coarse-grain techniques and Monte Carlo which offer further advances into molecular dynamics. As computational methods advance they will have more application by revealing the very subtle interactions that take place between the lipid and protein components of membranes - and which are so essential to their function. PMID- 16214219 TI - Oculomotor studies of cerebellar function in autism. AB - Histopathological, neuroimaging and genetic findings indicate cerebellar abnormalities in autism, but the extent of neurophysiological dysfunction associated with those findings has not been systematically examined. Suppression of intrusive saccades (square wave jerks) and the ability to sustain eccentric gaze, two phenomena requiring intact cerebellar function, were examined in 52 high-functioning individuals with autism and 52 age- and IQ-matched healthy subjects during visual fixation of static central and peripheral targets. Rates of intrusive saccades were not increased in autism during visual fixation, and foveopetal ocular drift was also not increased when subjects held an eccentric gaze. The absence of gross disturbances of visual fixation associated with cerebellar disease in individuals with autism, such as increased square wave jerk rates and foveopetal drift when holding eccentric gaze, indicates that the functional integrity of cerebellar--brainstem networks devoted to oculomotor control is preserved in autism despite reported anatomic variations. However, increased amplitude of intrusive saccades and reduced latency of target refixation after intrusive saccades were observed in individuals with autism, especially when subjects maintained fixation of remembered target locations without sensory guidance. The atypical metrics of intrusive saccades that were observed may be attributable to faulty functional connectivity in cortico cerebellar networks. PMID- 16214220 TI - Event-related potentials elicited by distractors in an auditory oddball paradigm in schizophrenia. AB - Patients with schizophrenia are affected more adversely than healthy controls by distracting conditions, due to their inability to adequately apportion attentional resources to targets or distractors. We attempted to re-evaluate the effects of distractors in 25 patients with chronic schizophrenia and in 12 controls. They performed an auditory target-detection task with 1500 Hz tone distractors and an additional control condition where a 1500-Hz tone was used as the target. The rate of target misses for patients with schizophrenia was 3.79% in non-distractor conditions and 14.79% in distractor conditions. Significantly reduced N100 responses to distractors and distractor condition targets were found. P300 responses to all target stimulus categories were reduced, but P300 responses to distractors were equal to those in the control group. There was a reduction of P300 amplitudes to distractors in both groups; however, only the control group showed significant enlargement of P300 amplitude when the distractors became the target stimuli. There is evidence that patients with schizophrenia tend to be less able to allocate their attentional resources adequately to target vs. distractor stimuli. When the distractors became the target stimuli, their responses remained unchanged, which suggests their inability to appropriately integrate stimulus information with contextual information. PMID- 16214222 TI - Alpha1-antitrypsin inhibits Moraxella catarrhalis MID protein-induced tonsillar B cell proliferation and IL-6 release. AB - Alpha1-antitrypsin (AAT) is a major circulating and tissues inhibitor of serine proteinases implicated in the regulation of inflammation and host defence. There is now increasing evidence that AAT may also exhibit anti-inflammatory activities independent of its protease inhibitor function. This study was undertaken to investigate the effects of native (inhibitory) and polymerized (non-inhibitory) forms of AAT on MID (Moraxella IgD binding protein)-induced human tonsillar B cell activation in vitro. We found that 0.5 microg/ml MID induces B cell proliferation and stimulates IL-6 release (p<0.001) relative to non-stimulated controls. Both native and polymerized AAT (0.5 mg/ml) inhibited MID-stimulated B cell proliferation in a similar manner (by 70%, p<0.001), whereas MID-induced IL 6 release was more strongly suppressed by polymerized (9.9-fold, p<0.001) as compared to native AAT (2.8-fold, p<0.01). Electrophoretic analysis of cell culture media did not indicate any interaction between AAT and MID, and flow cytometry data showed no competition for the same receptor. The effects of AATs were observed whether added together with MID or 2h after MID-addition to cell cultures. Thus, our data demonstrate that AAT inhibits MID-induced B cell activation in vitro that is unrelated to its protease inhibitory activity and is not dependent on MID binding to the cell surface. PMID- 16214221 TI - The selective inhibition of nitric oxide production in the avian macrophage cell line HD11. AB - The production of reactive nitrogen, nitric oxide (NO), has previously been demonstrated to be a major mechanism by which the innate immune system defends against microbial invasion. The induction of many antimicrobial mechanisms is regulated by numerous components during the transduction of the signal from the cell surface to the cell nucleus where response genes are upregulated. Toll-like cell surface receptor activation often leads to sequential modulation of protein tyrosine kinases (PTK), mitogen activated protein kinases (MAPK), degradation of I kappa B (IkappaB) regulatory molecules which, in turn, release the nuclear factor-kappa B (NF-kappaB) family proteins for translocation into the nucleus and subsequent gene transcription. The purpose of this study was to investigate components of the upstream signal transduction pathway induced by bacterial and viral-like stimulation of NO for antimicrobial defense by the transformed chicken macrophage cell line, HD11. We quantified the production of nitrite by chicken macrophages after exposure to selective pharmacological inhibitors of specific signal transduction components prior to stimulation by polyinosinic polycytidylic acid (poly I:C), formalin-fixed Enterococcus gallinarum (EG) or formalin-fixed Klebsiella pneumoniae (KP). We found that NO production induced by dsRNA or bacteria was reduced in a dose dependent manner by specific inhibitors of PTK, p38 MAPK, IkappaB, and NF-kappaB. Inhibition efficacy varied dependent on stimulation by bacterial or viral-like ligands. In general, NO production induced by bacterial stimulation was most effectively reduced by inhibition of p38 MAPK and least effectively reduced by inhibition of IkappaB. NF-kappaB and IkappaB inhibition affected NO production induced by dsRNA more than that induced by bacterial stimulation. PMID- 16214223 TI - Altered expression of chemokine receptor CXCR5 on T cells of myasthenia gravis patients. AB - Myasthenia gravis (MG) is characterized by the T cell-dependent production of anti-acetylcholine receptor (AChR) antibodies. The chemokine receptor CXCR5 regulates lymphocyte migration and is expressed on a subset of CD4+ T cells named follicular helper T cells (T(FH)), the key modulators of antibody production by B cells. We studied the frequency of CXCR5-positive lymphocytes in the peripheral blood of MG patients before and after therapy (thymectomy plus glucocorticoid). Before therapy, the MG patients showed a significantly higher frequency of CXCR5+ CD4+ T cells in the peripheral blood compared with the control group, while no significant difference in the percentages of CXCR5+ CD4+ T cells was observed between the patients of the hyperplasia group and those of the thymoma group. The CXCR5+ CD4+ T cell frequency correlated with the disease severity. The CXCR5+ CD4+ T cell frequency of MG patients positive for other autoantibodies together with anti-AChR antibodies was significantly higher than in those having only anti AChR antibodies. After therapy, the CXCR5+ CD4+ T cell percentage decreased gradually to the control level with a significant inverse correlation between the CXCR5+ CD4+ T cell frequency and duration after the initiation of MG therapy. The CXCR5+ CD4+ T cell populations in the hyperplastic thymuses and thymomas were not significantly different from those in the control thymuses. These results suggest that CXCR5+ CD4+ T cells play an important role in the disease activity of MG and that some MG patients have a systemic abnormality in T cell-dependent antibody production. PMID- 16214224 TI - The human PNMA family: novel neuronal proteins implicated in paraneoplastic neurological disease. AB - Using sera from patients with paraneoplastic neurological syndromes, several novel neuronal autoantigens such as the paraneoplastic Ma antigens (PNMA) have been identified. Here, we report the correction and completion of the previously published prototype member PNMA1, the brain and testis restricted expression of a third member (PNMA3) and the sequences for further partially uncharacterized members of this novel neuronal protein family. Murine and rat orthologs exist for this protein family. By analogy to the pro-apoptotic MOAP1, similar functional interactions may exist between members of the PNMA family and the bcl-2 family. PMID- 16214225 TI - Hearing loss as a sequel of secretory and acute otitis media as reflected by audiometric screening of Swedish conscripts. AB - OBJECTIVE: Many authors have shown a small effect on hearing from middle ear disease even a long time after active disease has subsided. We wanted to see to what extent those who have had otitis media in childhood in our district have normal hearing at the age of 18 years. The aim of the present study is to find out if a history of secretory otitis media (SOM) up to 14 years of age or acute otitis media (AOM) up to 4 years of age affects hearing at conscription in an age cohort of Swedish boys. METHODS: For a cohort of 951 boys we have good knowledge of their ear diseases up to 14 years of age from previous studies. The prevalence of elevated thresholds in audiometric screening at conscription was compared for the group of boys who had been treated with tympanostomy tubes and the group of boys with no history of ear disease. Similarly, boys with a history of ear disease but no treatment with tubes were compared to boys without history of ear disease. We also compared the prevalence of elevated thresholds for boys with a history of at least four episodes of AOM up to the age of four to those who had had no episodes up to that age. RESULTS: Those who had been treated with tympanostomy tubes because of longstanding SOM had a significantly higher prevalence of elevated thresholds for solitary frequencies. Two treated boys had a pure tone average greater than 25 dB, one unilaterally, one bilaterally. No difference could be seen between those who had had episodes of ear disease but no treatment with tympanostomy tubes and those who had had no documented ear disease. The prevalence of elevated thresholds was not correlated to more than three treated episodes of acute otitis media up to the age of 4 years. CONCLUSIONS: Boys who had been treated with tympanostomy tubes had a higher prevalence of having a hearing threshold above 20 dB for at least one frequency at the age of 18. Hearing handicap was not a common sequel to SOM. We can show no relation between more than three treated episodes of AOM before the age of four and screening results at the age of 18. PMID- 16214226 TI - Observations in a cohort of infants with severe laryngeal dyskinesia Auditory brainstem response may aid in the diagnosis. AB - OBJECTIVES: Laryngeal dyskinesia is not a well defined clinical entity, and difficult to distinguish from some other laryngeal disorders. We report on the observations, especially auditory brainstem responses, in a series of infants with laryngeal dyskinesia. METHODS: All infants with laryngeal dyskinesia, diagnosed in Sophia Children's Hospital between 1 January 2000 and 31 June 2003, were included in this retrospective case series study. Auditory brainstem responses were available in most infants. The respiratory, neurological and other symptoms, were recorded. RESULTS: Nine infants were diagnosed with laryngeal dyskinesia. Eight demonstrated severely abnormal auditory brainstem responses (ABR), indicating brainstem dysfunction. Most infants also had serious respiratory, and/or neurological disorders. DISCUSSION: It is hypothesized that laryngeal dyskinesia is a manifestation of a complex neurological disorder localized in the brainstem, comparable with brainstem dysgenesis syndrome. We suggest to use ABR in the diagnosis of unexplained laryngeal obstructive disorders to better define the pathology, and possibly the etiology of the disease. PMID- 16214227 TI - Rhino-bronchial syndrome in children: pathogenic correlations and clinical experimental aspects. AB - OBJECTIVE: This study aims at defining the incidence of rhino-bronchial syndrome (RBS) in children in order both to verify the influence of nasal obstructions on the disease and to determine therapeutic strategies which may cure the syndrome effectively at its early stage. METHODS: The investigation includes 128 non allergic children with obstructive disorders (adenoid hypertrophy, septal deviation, etc.) and rhino-sinus inflammations associated with bronchopulmonary diseases (asthma, chronic cough, bronchopulmonary infections). Medical and/or surgical treatment was chosen in consideration of the type and entity of the patients' main nasal pathology. At least 1 year follow-up was provided for each case to establish the improvement in the disorders affecting both the lower and upper airways. The results were statistically assessed. RESULTS: Medical and mainly surgical treatment always cured the upper airways disorders in patients with chronic nasal obstruction and rhino-sinus inflammation. Improvement of bronchopulmonary disease was reported in about half of the patients (49.4%). Statistically significant results were obtained only in the group with recurrent bronchopulmonary infections (80.9%, p<0.05). CONCLUSIONS: The study confirms that RBS may be quite frequent in childhood. This disorder has a multifactorial pathogenesis prevalently due to nasal obstruction and rhino-sinus infections. In the population studied, among the lower airways disorders, only infective bronchopulmonary inflammation showed a significant correlation in the assessment between lower and upper airways disorders. In order to prevent the progression of the syndrome to serious pathologic events of the lower airways, a prompt and effective treatment of children's nasal disorders is thus recommended. PMID- 16214228 TI - Development and application of a quantitative RT-PCR potency assay for a pentavalent rotavirus vaccine (RotaTeq). AB - A sensitive and reproducible method to determine the in vitro infectious potency of a pentavalent reassortant rotavirus vaccine (RotaTeq) has been developed as an alternative to classical potency assays. Potency was determined based on cell based viral replication followed by quantitative reverse-transcription polymerase chain reaction (RT-QPCR) analysis. In the assay, confluent Vero cell monolayers in 96-well plates were inoculated with serial dilutions of test samples, a pentavalent reassortant rotavirus reference standard and assay controls, followed by incubation for 24h. The cells were lysed with a Triton X-100 solution and the lysates assayed by RT-QPCR to quantitate viral nucleic acid produced during replication. The RT-QPCR utilizes primer/probe sets specific to each virus reassortant and the potencies of each sample were determined relative to the reference standard. This assay, hereafter referred to as the Multivalent QPCR Based Potency Assay (M-QPA), permits the specific quantitation of each individual reassortant virus in the presence of the other four reassortant viruses. In addition, the assay was demonstrated to be concordant with a traditional method (plaque assay) for the quantitation of infectious virus particles. It is anticipated that assays of this type will become a valuable tool in the assignment of potency values and in the monitoring of stability of live virus vaccines. PMID- 16214229 TI - Sensitive and rapid detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) in shrimps by loop-mediated isothermal amplification. AB - A method for nucleic acid amplification, loop-mediated isothermal amplification (LAMP) is a novel, sensitive and rapid technique, which can be applied for disease diagnosis in aquaculture. Using the LAMP method, a highly specific and sensitive diagnostic system for infectious hypodermal and hematopoietic necrosis virus (IHHNV) detection was designed. A set of four primers was designed by targeting the IHHNV genome DNA. By the detection system, target DNA was amplified and visualized on agarose gel within 60min under isothermal condition at 64 degrees C. Without gel electrophoresis, the LAMP amplicon was visualized directly in the reaction tube by addition of SYBR Green I for a naked-eye inspection. The LAMP reaction was also assessed by the white turbidity of magnesium pyrophosphate (a by-product of LAMP) in the tube. The assay had a detection limit of 5-500 copies of DNA template with gel electrophoresis, SYBR Green I and white turbidity with naked-eye inspection. The detection sensitivity of LAMP was 100-fold higher than the PCR. A diagnostic procedure which is rapid and highly sensitive was developed for IHHNV detection. PMID- 16214230 TI - A neuroendocrine mechanism for sustaining fear. AB - Fear is an adaptive response to recognition of a potentially dangerous event. Glucocorticoids are essential for maintaining a wide variety of behavioral events by their regulation of numerous genes; one such gene encodes corticotrophin releasing hormone (CRH). CRH is involved in diverse behavioral responses to changing environmental demands. In this review, we focus on one aspect of glucocorticoid regulation of CRH--namely, fear-related responses to diverse classes of adverse events, such as those represented by contextual and cue specific stimuli. Three extra-hypothalamic forebrain sites appear crucial for fear-related behavioral responses: the amygdala and the bed nucleus of the stria terminalis for sustaining adaptive fear-related behaviors, and the medial prefrontal cortex for modulating fear-related behaviors. Central regulation of CRH by glucocorticoids is important for adaptive and sustained fear-related behaviors, and its aberration is associated with anxiety and depressive disorders. PMID- 16214231 TI - Polymer-bound 6' sialyl-N-acetyllactosamine protects mice infected by influenza virus. AB - To develop a mouse model for testing receptor attachment inhibitors of human influenza viruses, the human clinical virus isolate in MDCK cells A/NIB/23/89M (H1N1) was adapted to mice by serial passaging through mouse lungs. The adaptation enhanced the viral pathogenicity for mice, but preserved the virus receptor binding phenotype, preferential binding to 2-6-linked sialic acid receptors and low affinity for 2-3-linked receptors. Sequencing of the HA gene of the mouse-adapted virus A/NIB/23/89-MA revealed a loss of the glycosylation sites in positions 94 and 163 of HA1 and substitutions 275Asp-->Gly in HA1 and 145Asn- >Asp in HA2. The four mouse strains tested differed significantly in their sensitivity to A/NIB/23/89-MA with the sensitivity increasing in the order of BALB/cJCitMoise, C57BL/6LacSto, CBA/CaLacSto and A/SnJCitMoise strains. Testing of protective efficacy of the polyacrylamide conjugate bearing Neu5Acalpha2 6Galbeta1-4GlcNAc trisaccharide under conditions of lethal or sublethal virus infection demonstrated a strong protective effect of this preparation. In particular, aerosol treatment of mice with the polymeric attachment inhibitor on 24-110 h after infection completely prevented mortality in sensitive animals and lessened disease symptoms in more resistant mouse strains. PMID- 16214232 TI - Influence of estrus cycle and ageing on activity patterns in two inbred mouse strains. AB - Despite the widespread use of inbred mice in research, little is known about aging of the circadian system in female mice, although interactions between female gonadal hormones and circadian rhythms have been established. We investigated the influence of the estrus cycle on circadian aspects of running wheel activity and changes in the course of aging in female C57BL/6 and C3H/He mice recorded continuously between the ages of 3 and 19 months. In the young, cycling mice the second part of the proestrus night was often, but not consistently, characterized by increased motor activity compared to the remaining estrus cycle nights. After estrus cycling had ceased in the course of ageing, the estrus-dependent day-to-day variability in activity was reduced. The amplitude of the daily rest-activity rhythm decreased progressively after the age of 8 months in C3H/He and 10 months in C57BL/6 mice. The capacity for resynchronisation of activity onset to the LD-cycle was compared in young and old mice after an 8-h phase advance of the LD-cycle. Resynchronisation was significantly slower in old C3H/He mice and unaffected by age in C57BL/6 mice. The circadian period in constant darkness did not change with age in either strain. However, the period was shorter in 17-month old C57BL/6 mice compared to an additional group, which was recorded at the same age, after at least 1-month adaptation to the recording conditions. The results show that the reproductive state as well as ageing influence motor activity patterns of female mice in a strain- and cohort dependent manner. PMID- 16214233 TI - Role of mu-opioid and NMDA receptors in the development and maintenance of repeated swim stress-induced thermal hyperalgesia. AB - Repeated exposure to swimming stress induces a long-lasting hyperalgesia in the rat by mechanisms to be elucidated. Since opioid and glutamate neurotransmitter systems modulate pain, we now evaluated the effect of pharmacological blockade of opioid and glutamate receptors subtypes on forced swimming stress-induced hyperalgesia. Male rats were daily subjected to 10-20 min of forced or sham swimming for 3 days and thermal nociception was estimated twice, before each behavioral conditioning and 24 h after the last, using hot plate test. Selective opioid and NMDA receptor antagonists were administered i.p. either before each conditioning session or before the second nociception assessment. Unlike sham swimming rats, forced swimming rats showed significant reductions in hot plate response latencies (hyperalgesia) after the last swimming session, as compared to pre-stress values. Rats treated with the opioid receptor antagonists naloxone (0.1 mg/kg, non-subtype-selective) and naloxonazine (5 mg/kg, mu(1)-subtype selective), before each forced swimming, did not become hyperalgesic, whereas those treated before the second post-stress assessment of nociception developed hyperalgesia. Naltrindole (0.5 mg/kg, delta-subtype-selective) and nor binaltorphimine (0.5mg/kg, kappa-subtype-selective) were inactive in both administration schedules. The efficacy of morphine (3-7.5 mg/kg) to produce analgesia in forced swimming rats was lower than in sham swimming rats. Rats treated with the NMDA antagonist ketamine (5 mg/kg) before the forced swimming or the second post-stress assessment of nociception did not have hyperalgesia. Thus, swim stress-induced hyperalgesia might be initiated by the repeated stimulation of mu-opioid and NMDA receptors but maintained only by the activity of NMDA receptors. PMID- 16214234 TI - Peripubertal anxiety profile can predict predisposition to spatial memory impairments following chronic stress. AB - We tested the hypothesis that peripubertal anxiety levels are predictive of the detrimental effects of chronic stress on hippocampal-dependent spatial memory. The anxiety levels of peripubertal male Sprague-Dawley rats (43 days old) were characterized using open field and elevated plus mazes, followed by chronic restraint stress for 6 h/day/21 days beginning in young adulthood (75 days). Following chronic stress treatment, rats were tested on the spatial Y-maze using two inter-trial interval levels of difficulty (4 h: 1 day post-chronic stress; 1 min: 2 days post-chronic stress). As expected, all groups displayed intact spatial memory in the less difficult 1 min version of the Y-maze. However, in the 4 h version of the Y-maze, chronically stressed high anxiety rats showed impaired spatial memory, while chronically stressed low anxiety and control (low and high anxiety) rats displayed intact spatial memory. Moreover, a month after chronic stress ended, high anxiety rats had significantly higher basal corticosterone levels than low anxiety rats (control and stress). These results indicate that peripubertal anxiety and chronic stress interact to influence hippocampal dependent spatial memory in adulthood. PMID- 16214235 TI - Individual differences in novelty-seeking behavior but not in anxiety response to a new environment can predict nicotine consumption in adolescent C57BL/6 mice. AB - Considering that adolescence is associated with an increased motivation to seek out new stimuli and with low anxiety levels in exploring novel environments, and that both behavioral traits may be associated with substance abuse, we investigated whether the behavioral response to a novel environment can predict subsequent oral nicotine self-administration in adolescent C57BL/6 mice. On postnatal day 30 (PN30), the novelty-seeking behavior and anxiety levels were assessed in a hole board activity box. The total number of head-dips (DIP) was used to classify animals either into the high novelty (HN; DIP above median) or low novelty (LN; DIP below median) groups. The percentage of center squares crossed (CEN) was used to classify animals either into the high anxiety (HA; CEN below median) or low anxiety (LA; CEN above median) groups. From PN31 to PN41, all animals were given a free choice between tap water or a nicotine solution (10 microg/ml). LN mice did not change nicotine intake throughout the free choice procedure, however, HN mice presented a marked increase in consumption. There were no differences in consumption between HA and LA mice. Our results indicated that mice that presented a more intense novelty-seeking behavior increased their preference for nicotine during the free choice experiment but that anxiety levels did not predict nicotine consumption. These results suggest that higher motivation to seek out new experiences is a significant contributor to drug use in adolescents and that anxiety is probably not a major factor that determines differential nicotine consumption during adolescence. PMID- 16214236 TI - Effects of (Z)-3-hexenol, a major component of green odor, on anxiety-related behavior of the mouse in an elevated plus-maze test and biogenic amines and their metabolites in the brain. AB - In the present study, the effect of (Z)-3-hexenol, one of the main constituent of green odor, on the anxiety-related behavior of mouse and the content of 5 hydroxytryptamine (5-HT), dopamine and their metabolites in the brain were investigated. Evaluation of anxiety-related animal behavior was performed by measuring the percent of time spent on the open arms and the percent of open-arm entries as conventional anxiety indices. The number of times displaying risk assessment from a closed arm was also measured as an ethological anxiety index. Diazepam, an anxiolytic agent, enhanced the percent of time spent on the open arms and the percent of open-arm entries. The number of times displaying risk assessment was not affected by diazepam in the present study. The percent of time on the open arms were depressed, and the number of times displaying risk assessment were stimulated by 1-(3-trifluoromethylphenyl)piperazine (TFMPP), indicating that TFMPP acts as an anxiogenic agent. (Z)-3-Hexenol revealed anxiolytic activity and increased the percent of time spent on the open arms and decreased the number of times displaying risk-assessment. In the neurochemical study, diazepam had no effect on the content of 5-HT and its metabolite in the brain cortex or hippocampus. On the other hand, TFMPP inhibited the 5-HT turnover rate accompanied by the elevation of the 5-HT content and reduction of the 5-HIAA content in the brain cortex and hippocampus. (Z)-3-Hexenol significantly increased the 5-HT content without affecting the 5-HIAA content or the 5-HT turnover rate in the brain cortex or hippocampus. Changes in serotonergic activity in the cortex and hippocampus were suggested to be involved in the anxiolytic effect of (Z)-3-hexenol observed in the elevated plus-maze test. PMID- 16214237 TI - Alpha-melanocyte stimulating hormone reduces putative stress-induced sickness behaviors in isolated guinea pig pups. AB - We have proposed that passive responses observed following maternal separation in guinea pig pups represent "stress-induced sickness behaviors" mediated by proinflammatory processes. In this study, the anti-inflammatory peptide, alpha melanocyte stimulating hormone (alpha-MSH) administered intracerebroventricularly, but not intraperitoneally, reduced the passive responses of crouching, eye-closing, and extensive piloerection relative to levels following administration of vehicle. These findings support our hypothesis and are as would be expected if pro-inflammatory processes act centrally to promote the passive behaviors of separated guinea pig pups. PMID- 16214238 TI - Hippocampal cell proliferation across the day: increase by running wheel activity, but no effect of sleep and wakefulness. AB - The present study investigated whether proliferation of hippocampal progenitors is subject to circadian modulation. Mice were perfused using 3h intervals throughout the light-dark cycle and brains were stained for Ki-67. Since Ki-67 is not expressed during the G0 phase of the cell cycle, we expected a decline in Ki 67 expression at the moment cells synchronously exit the cell cycle. However, despite the fact that various hippocampal factors fluctuate across the day, the number of dividing cells remained constant. In a second experiment, we studied whether disturbance of normal sleep affected the stable rate in cell proliferation. Our data show that 12h of sleep deprivation during the light phase did not influence proliferating cell number. A third experiment investigated whether physical activity, a condition known to enhance hippocampal cell proliferation, caused an elevation of the steady baseline number of proliferating progenitors, or a peak directly following the active phase of the animals. Mice were housed with a running wheel for 9 days. On the last day, animals were sacrificed either directly before or directly after the active phase. Exercise significantly promoted cell proliferation and this effect appeared to be strongest directly after the active period and to disappear during the resting phase. Our data suggest that hippocampal cell proliferation is not synchronized under basal conditions and is unchanged by sleep deprivation. However, running affected cell proliferation differentially at two times of day. These data demonstrate that the steady rate in cell proliferation is not indispensable, but can be changed by behavioral activity. PMID- 16214239 TI - The role of the hippocampus in object recognition in rats: examination of the influence of task parameters and lesion size. AB - Studies examining the effects of hippocampal lesions on object recognition memory in rats have produced conflicting results. The present study investigated how methodological differences and lesion size may have contributed to these discrepancies. In Experiment 1 we compared rats with complete, partial (septal) and sham hippocampal lesions on a spontaneous object recognition task, using a protocol previously reported to result in deficits following large hippocampal lesions . Rats with complete and partial hippocampal lesions were unimpaired, suggesting the hippocampus is not required for object recognition memory. However, rats with partial lesions showed relatively poor performance raising the possibility that floor effects masked a deficit on this group. In Experiment 2, we used a second spontaneous object recognition protocol similar to that used by the two other studies that have reported deficits following hippocampal lesions . Rats with complete hippocampal lesions were significantly impaired, whereas rats with partial lesions were unimpaired. However, the complete lesion group showed less object exploration during the sample phase. Thus, the apparent recognition memory deficit in Experiment 2 may be attributable to differential encoding. Together, these findings suggest that the hippocampus is not required for intact spontaneous object recognition memory. These findings suggest that levels of object exploration during the sample phase may be a critical issue, and raise the possibility that previous reports of object recognition deficits may be due to differences in object exploration rather than deficits in object recognition per se. PMID- 16214240 TI - alpha-Tocopherol and ascorbic acid prevent memory deficits provoked by chronic hyperprolinemia in rats. AB - In the present study we investigated the action of alpha-tocopherol and ascorbic acid on the effects elicited by chronic hyperprolinemia on rat performance in the Morris water maze. Rats received subcutaneous injections of proline (experimental group) twice a day, with 10 h-interval, from the 6 to 28th days of age or an equivalent volume of 0.9% saline solution (controls). Half of the proline-treated group also received intraperitoneal administration of alpha-tocopherol (40 mg/kg) and of ascorbic acid (100 mg/kg) from the 6 to 28th days of life. On the 60th day of life, rats were subjected to testing in the water maze. Results show that chronic proline administration provokes impairment on spatial learning in reference memory task, as revealed by the increase of latency in acquisition, in the probe trial and in crossing over the platform location, as well as by the number of crossings, when compared to saline-treated animals. Proline-treated rats also demonstrated a reduced efficiency to find the platform position in the working memory task. Rats chronically treated with proline plus alpha-tocopherol and ascorbic acid had above effects prevented, suggesting the participation of oxidative stress in such effects. Our findings lend support to a novel therapeutic strategy, based on these vitamins, to the cognitive dysfunction associated with hyperprolinemia type II. PMID- 16214241 TI - Antinociceptive role of galanin in the spinal cord of rats with inflammation, an involvement of opioid systems. AB - The present study investigated the role of galanin in the transmission of nociceptive information in the spinal cord of rats with inflammation. Bilateral decreases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation were observed after acute inflammation induced by injection of carrageenan into the plantar region of the rat left hindpaw. Intrathecal injection of galanin induced significant increases in the HWLs to thermal and mechanical stimulation in rats with inflammation. The galanin-induced antinociceptive effect was more pronounced in rats with inflammation than that in intact rats. The antinociceptive effect of galanin was partly inhibited by intrathecal injection of naloxone. Furthermore, intrathecal administration of galantide, an antagonist of galanin receptor, could attenuate the antinociceptive effect induced by intraperitoneal injection of morphine, suggesting an involvement of opioid systems in the galanin-induced antinociception. The results indicate that galanin plays an important role in the transmission of nociceptive information in the spinal cord of rats with inflammation, and opioid systems are involved in the galanin-induced antinociception. PMID- 16214242 TI - Evaluation of rage isoforms, ligands, and signaling in the brain. AB - Since the identification of the receptor for advanced glycosylation end products (RAGE) in 1992, there have been tremendous strides made in our understanding of the role RAGE receptors play in a variety of physiological and pathological processes. Despite such progress, several fundamental aspects of RAGE expression and RAGE function remain largely unanswered. In particular, while multiple forms of the RAGE receptor are known to exist, little is known with regards to how these different isoforms of the RAGE receptor work together to mediate RAGE signaling. For example, some forms of the RAGE receptor may promote deleterious feed-forward pathways, while others may serve to inhibit deleterious activation of the RAGE receptor. Additionally, important questions remain with regards to the intracellular domain of the full-length RAGE receptor, and the specifics surrounding how intracellular signaling pathways become activated via the RAGE family of receptors. The focus of this review is to address each of these important issues, as well as other key aspects of RAGE biology, and discuss how they are important for both our understanding of the physiological and pathological roles of RAGE signaling within the brain. PMID- 16214243 TI - Structure of caveolae. AB - The introduction of the electron microscope to the study of the biological materials in the second half of the last century has dramatically expanded our view and understanding of the inner workings of cells by enabling the discovery and study of subcellular organelles. A population of flask-shaped or spherical invaginations of the plasma membrane were described and named plasmalemmal vesicles or caveolae. Until the discovery of caveolin-1 as their first molecular marker in early 1990s, the study of caveolae was the exclusive domain of electron microscopists that demonstrated caveolae at different surface densities in most mammalian cells with few exceptions. Electron microscopy techniques in combination with other approaches have also revealed the structural features of caveolae as well as some of their protein and lipid residents. This review summarizes the data on the structure and components of caveolae and their stomatal diaphragms. PMID- 16214244 TI - Oxidation of serum low-density lipoprotein (LDL) and antioxidant status in young and elderly humans. AB - The incidence of atherosclerosis increases with age, as do various indices of free-radical mediated damage, e.g., lipid peroxidation. Because lipid peroxidation plays a prominent role in lipoprotein oxidation, likely a prelude to atherosclerosis, we compared the susceptibility of lipoproteins to oxidation in young (19-30 years) and elderly (59-86 years) groups. Although we found no significant differences in serum malondialdehyde (MDA) or oxidized LDL antibodies (OLAB) between young and elderly lipoproteins, MDA was directly related to OLAB regardless of age (r = 0.322, p = 0.005) and there was a trend for lower OLAB levels (30.5%, p = 0.079) in the elderly compared to the young population. Overall, serum antioxidant status was either similar or greater in the elderly group compared to the young group, likely reflecting antioxidant supplementation by the elderly group. OLAB was inversely related to Vitamin C (r = -0.310, p = 0.008) and Vitamin E intake (r = -0.277, p = 0.018) from foods and supplements. Serum levels of Vitamin C and Vitamin E were significantly higher (18.5%, p = 0.021 and 58.1 %, p < 0.001, respectively) in the elderly group compared to the young group and the ratio of Vitamin E to Vitamin C was significantly higher (30.4%, p = 0.042) in the serum of the elderly group. Oxidation of serum LDL and antioxidant status were not affected by age; however, the ratio of serum Vitamin E to Vitamin C was higher in the elderly group which may affect Vitamin E recycling. PMID- 16214245 TI - Self-rated health and mortality: could clinical and performance-based measures of health and functioning explain the association? AB - It is well established that self-rated health (SRH) predicts mortality even when other indicators of health status are taken into account. It has been suggested that SRH measures a wide array of mortality-related physiological and pathological characteristics not captured by the covariates included in the analyses. Our aim was to test this hypothesis by examining the predictive value of SRH on mortality controlling for different measurements of body structure, performance-based functioning and diagnosed diseases with a population-based, prospective study over an 18-year follow-up. Subjects consisted of 257 male residents of the city of Jyvaskyla, central Finland, aged 51-55 and 71-75 years. Among the 71-75-year-olds the association between SRH and mortality was weaker over the longer compared to shorter follow-up period. In the multivariate Cox regression models with an 18-year follow-up time for middle-aged and a10-year follow-up time for older men, SRH predicted mortality even when the anthropometrics, clinical chemistry and performance-based measures of functioning were controlled for, but not when the number of chronic diseases was included. Although our results confirm the hypothesis that the predictive value of SRH can be explained by diagnosed diseases, its predictive power remained, when the clinical and performance-based measures of health and functioning were controlled. PMID- 16214246 TI - Psychological abuse and its characteristic correlates among elderly Taiwanese. AB - This study examined the prevalence of psychological abuse and identified individual characteristics associated with different levels of psychological abuse in a group of randomly selected elderly Taiwanese. A cross-sectional design was performed that utilized a descriptive-correlational approach. Data for this study were gathered from several communities in southern Taiwan. The sample comprised 195 elderly Taiwanese aged 60 years or older, capable of verbal communication, and partially dependent on a caregiver. The Psychological Elder Abuse Scale (PEAS), the Short Portable Mental State Questionnaire (SPMSQ), and the Barthel's Index were used for data collection. The findings demonstrated that the PEAS score range was 0-24 (mean 6.32 +/- 4.6). Moreover, 44 subjects (22.6%) had scores of ten or greater. The most frequently reported indicators in the PEAS were "wishes to see family members unfulfilled" and "economic dependence on others". The analytical results also showed that the SPMSQ and Barthel's Index scores were strongly related to the PEAS score (p < 0.001), indicating psychological abuse appeared higher among elderly people with lower cognitive and physical functioning. The findings of this study are crucial in developing a national system for protecting the elderly, and a continuing educational plan is required for enforcing communication between caregivers and elders. PMID- 16214247 TI - Top of the charts: download versus citations in the International Journal of Cardiology. AB - The medical literature is growing at an alarming rate. Research assessment exercises, research quality frameworks, league tables and the like have attempted to quantify the volume, quality and impact of research. Yet the established measures (such as citation rates) are being challenged by the sheer number of journals, variability in the "gold standard" of peer-review and the emergence of open-source or web-based journals. In the last few years, we have seen a growth in downloads to individual journal articles that now easily exceeds formal journal subscriptions. We have recorded the 10 top cited articles over a 12-month period and compared them to the 10 most popular articles being downloaded over the same time period. The citation-based listing included basic and applied, observational and interventional original research reports. For downloaded articles, which have shown a dramatic increase for the International Journal of Cardiology from 48,000 in 2002 to 120,000 in 2003 to 200,000 in 2004, the most popular articles over the same period are very different and are dominated by up to-date reviews of either cutting-edge topics (such as the potential of stem cells) or of the management of rare or unusual conditions. There is no overlap between the two lists despite covering exactly the same 12-month period and using measures of peer esteem. Perhaps the time has come to look at the usage of articles rather than, or in addition to, their referencing. PMID- 16214248 TI - Characteristics in the epidemiology of bovine viral diarrhea virus (BVDV) of relevance to control. AB - An understanding of the driving forces of BVDV transmission can be gained by considering the reproductive rate, between individuals and between herds. The former determines the prospects for eliminating the infection from herds, and the latter is the key to persistence at the population level. In this paper, the relation between these two characteristics, their underlying parameters and measures and priorities for BVDV control are discussed. A general model for BVDV control is outlined, with bio-security, virus elimination and monitoring as three necessary consecutive elements, and with immunization as an optional step. A distinction is made between systematic and non-systematic approaches to BVDV control (where the former refers to a monitored and goal-oriented reduction in the incidence and prevalence of BVDV infection and the latter to where measures are implemented on a herd-to-herd decision basis and without systematic monitoring in place). Predictors of progress for systematic control approaches in general are discussed in terms of the abilities: to prevent new infections, to rapidly detect new cases of infection, to take action in infected herds and to gain acceptance by stakeholders. We conclude that an understanding not only of the biology, but also of the social factors - human behavior, the motives that makes stakeholders follow advice and the cultural differences in this respect - are important factors in forming recommendations on alternative strategies for BVDV control. PMID- 16214249 TI - Indication of transmission of BVDV in the absence of persistently infected (PI) animals. AB - In a closed dairy herd all animals were tested serologically for BVD antibodies twice a year during a 6-year period. Seroconversions were detected every year. At the start of the 6-year monitoring period blood samples from all animals were examined by virus isolation. No persistently infected animals were identified. Entire-herd culturing for BVDV was repeated at the end of the third year. Samples from all newborn female calves were examined for BVDV at approximately 2 months of age and older. During the entire monitoring period BVDV was isolated in one newborn calf twice with an interval of 3 weeks. The mother had seroconverted during pregnancy. Five congenitally infected non-PI calves were identified, the mothers of which had seroconverted during late pregnancy; repeated sampling proved the calves to remain seropositive in a seronegative age cohort. Although direct and indirect introduction of BVDV from outside the herd can never be excluded it seems highly unlikely in this closed herd. The findings indicate that transmission of BVDV can take place over a long period of time in the absence of PI animals. This observation may have serious consequences for control programmes. PMID- 16214250 TI - Effects of pre-stimulus processing on subsequent events in a warned Go/NoGo paradigm: response preparation, execution and inhibition. AB - The cued Go/NoGo task elicits response preparation during the foreperiod, and, depending on the S2 signal, either response execution or inhibition. This study aimed to determine how processes in the foreperiod might affect or predict post S2 processing. Thirty-two adults participated in a cued Go/NoGo task (50% Go), with a median split of mean RT producing "Fast" and "Slow" groups. ERP measures were subjected to both ANOVA and regression techniques. There were no differences in the NoGo N2 effect related to response speed, nor was the effect related to pre-S2 processes. The anterior shift of the NoGo P3 was larger in the Fast group, and while the late CNV was associated with the absolute amplitude of both Go and NoGo P3, it was not related to the anterior-posterior Go/NoGo differences. Together, these data suggest that the inhibitory process may be reflected in the NoGo P3 effect, rather than the NoGo N2 effect. PMID- 16214251 TI - Characterization of acetic acid bacteria in "traditional balsamic vinegar". AB - This study evaluated the glucose tolerance of acetic acid bacteria strains isolated from Traditional Balsamic Vinegar. The results showed that the greatest hurdle to acetic acid bacteria growth is the high sugar concentration, since the majority of the isolated strains are inhibited by 25% of glucose. Sugar tolerance is an important technological trait because Traditional Balsamic Vinegar is made with concentrated cooked must. On the contrary, ethanol concentration of the cooked and fermented must is less significant for acetic acid bacteria growth. A tentative identification of the isolated strains was done by 16S-23S-5S rDNA PCR/RFLP technique and the isolated strains were clustered: 32 strains belong to Gluconacetobacter xylinus group, two strains to Acetobacter pasteurianus group and one to Acetobacter aceti. PMID- 16214252 TI - Improving recombinant MVA immune responses: potentiation of the immune responses to HIV-1 with MVA and DNA vectors expressing Env and the cytokines IL-12 and IFN gamma. AB - Recombinants based on vaccinia virus vectors, especially on the highly attenuated modified vaccinia virus Ankara (MVA) strain, are now being tested in clinical trials for safety and immunogenicity, using prime/boost heterologous regimes of vaccination. Due to the limited replication capacity of MVA, it is necessary to develop procedures that can enhance the specific cellular immune responses to the recombinant antigen delivered by the MVA vector. In this investigation, we have characterized the systemic immune responses in BALB/c mice using interferon-gamma (IFN-gamma) or interleukin-12 (IL-12) in an adjuvant-like manner elicited by MVA recombinants or naked DNA vectors expressing one of those cytokines in combination with the human immunodeficiency virus type 1 (HIV-1) envelope (Env) as antigen. In infected mice, virus gene expression in splenocytes and levels of cytokines IFN-gamma and IL-12 in serum were maximal by 6h post-infection (hpi) with MVA recombinants expressing IFN-gamma (MVAIFN-gamma) or IL-12 (MVAIL-12). In the infected animals, co-expression of HIV-1 env (MVAENV) and either IFN-gamma or IL-12 from MVA recombinants produced a two and three-fold increase of anti-env CD8+ T cell response, respectively. When priming was carried out with DNA vectors expressing HIV-1 env and either IFN-gamma or IL-12, the magnitude of the specific anti-env CD8+ T cell stimulation after MVAENV booster was further enhanced. Our findings revealed that IFN-gamma or IL-12 can be used to potentiate the cellular immune response to HIV-1 env, when delivered either from a single MVA recombinant or from a DNA vector. The increment of the CD8+ T cell response was higher in a DNA/MVA prime/boost protocol. Thus, the immune response of MVA vectors can be improved with the co-delivery of the cytokines IFN-gamma or IL-12. PMID- 16214253 TI - Patterns of sequence evolution at epitopes for host antibodies and cytotoxic T lymphocytes in human immunodeficiency virus type 1. AB - Analysis of published sequence data from the nine protein-coding genes of human immunodeficiency virus type 1 (HIV-1) showed striking differences in evolutionary pattern between epitopes for host neutralizing antibodies (Ab) and epitopes for cytotoxic T cells (CTL). In all sequences analyzed, the greatest median amino acid residue diversity was seen at sites that formed part of Ab epitopes, but not of CTL epitopes. By contrast, sites belonging to CTL epitopes but not to Ab epitopes showed reduced median amino acid sequence diversity not only in comparison to sites in Ab epitopes but also in comparison to non-epitope sites. Ab epitopes that did not overlap CTL epitopes showed the highest frequency of comparisons in which the rate of nonsynonymous (amino acid-altering) nucleotide substitution exceeded that of synonymous nucleotide substitution, supporting the hypothesis that much of the diversity at Ab epitopes results from positive selection exerted by the host immune system. Though less frequent than that at Ab epitopes, there was evidence of such selection at certain CTL epitopes as well; and amino acid differences between sister pairs of sequences in CTL epitopes were more likely to be convergent than those in Ab epitopes. The pattern seen at CTL epitopes may represent the result of conflicting pressures favoring conservation of the amino acid sequence for functional reasons and amino acid replacements for reasons of CTL escape. PMID- 16214255 TI - Beneficial effects of nateglinide on insulin resistance in type 2 diabetes. AB - Nateglinide, a rapid insulin secretagogue, is known to facilitate the early phase of insulin secretion and has been used for the treatment of type 2 diabetic patients with postprandial hyperglycemia. The aim of this study is to evaluate the effect of nateglinide on insulin resistance as well as insulin secretory defects in type 2 diabetic patients. Insulin secretion ability was evaluated by the hyperglycemic clamp test, and insulin sensitivity was evaluated by the euglycemic hyperinsulinemic clamp test, using an artificial pancreas. The hyperglycemic clamp test showed that a 7-day treatment with nateglinide significantly increased insulin secretion in response to high glucose. Interestingly, although nateglinide is known to facilitate insulin secretion, daily urinary C-peptide excretion was decreased after nateglinide treatment. Moreover, in the euglycemic hyperinsulinemic clamp test, glucose infusion rate was significantly increased by nateglinide treatment, indicating that nateglinide functions to decrease insulin resistance. Nateglinide ameliorates insulin resistance as well as insulin secretory defects in type 2 diabetic patients. PMID- 16214254 TI - Thermoresponsive polymers as gene delivery vectors: cell viability, DNA transport and transfection studies. AB - A range of gene delivery vectors containing the thermoresponsive polymer, poly(N isopropylacrylamide) (PNIPAm) was evaluated for effects on cell viability, intracellular trafficking and transgene expression in C2C12 mouse muscle cells. Polymers were complexed with plasmid DNA at pH 7.4 and the ability of the resulting particles to transfect cells was assessed via confocal microscopy and protein expression studies in tissue culture. Cell viability assays indicated that these polymers were toxic at high concentrations when not complexed to DNA or at certain polymer:DNA ratios. Poly(ethyleneimine) co-polymers with side-chain grafted PNIPAm were shown to be less toxic than poly(ethyleneimine) alone or PNIPAm-co-(N,N'-dimethylaminoethylmethacrylate) linear co-polymers and the effects were concentration dependent. Confocal micrographs of labeled polymers and DNA indicated rapid cellular entry for all the complexes but expression of Green Fluorescent Protein was achieved only when the branched PEI-PNIPAm co polymers were used as vectors. The results indicate that design of appropriate co polymer components and overall polymer architecture can be used to mediate, and perhaps ultimately control, DNA transport and transgene expression. PMID- 16214256 TI - Serum magnesium concentrations in type 1 diabetic patients: relation to early atherosclerosis. AB - Hypomagnesemia and sub-clinical atherosclerosis are common in type 1 diabetic patients, and are especially common in poorly controlled and chronically treated diabetics. The aim of this study was to evaluate the relationships between serum magnesium (Mg) and intima-media thickness (IMT), and functions of common carotid artery (CCA), accepted as markers of early carotid atherosclerosis in type 1 diabetic patients. Serum magnesium levels were measured in 51 diabetic patients (26 girls and 25 boys). Age, duration of diabetes as well as major cardiovascular risk factors, including anthropometric and metabolic parameters were matched between girls and boys. CCA structure and functions were measured by ultrasonography as IMT, cross-sectional compliance (CSC), cross-sectional distensibility (CSD), diastolic wall stress (DWS), and incremental elastic modulus (IEM). The difference for serum Mg was significant between diabetic girls and boys (0.75+/-0.09 mmol/l versus 0.86+/-0.14 mmol/l; p=0.02). Mg levels were correlated with IMT (r=-0.58, p=0.004) and DWS (r=-0.49, p=0.01) in girls and with IMT (r=-0.55, p=0.007) and DWS (r=-0.46, p=0.03) in boys. In a multivariate regression model, Mg, systolic blood pressure and waist-hip ratio emerged as independent correlates for mean IMT in girls with the total variance explained being 63%, and Mg and triglycerides emerged as independent correlates for mean IMT in boys with the total variance explained being 68%. Mg did not emerge as independent correlates for mean DWS in both girls and boys. Serum magnesium levels in children and adolescent patients with type 1 diabetes are associated with early atherosclerosis. PMID- 16214257 TI - The Common Drug Review: a NICE start for Canada? AB - Prescription drugs are one of the fastest growing cost components of modern health care systems. Efforts to control escalating costs while simultaneously maximizing population health outcomes have led many countries to implement restrictive criteria on the funding of certain drugs. While drugs are licensed for sale based on evidence of safety and efficacy versus a placebo, many funders now require evidence of clinical- and cost-effectiveness compared to existing drugs as part of their reimbursement criteria. In some countries, concerns about duplication of drug assessment and administrative effort across different jurisdictions have led to experimentation with various forms of centralized drug review processes. Centralized drug reviews strive to standardize, inform, and improve drug reimbursement decisions through critical assessments of comparative clinical- and cost-effectiveness. The ultimate objective is to inform formulary listing decisions that both maximize health outcomes and achieve good "value for money". This paper describes the Common Drug Review (CDR), a uniquely Canadian version of a centralized drug review process, and compares it with the much studied National Institute for Health and Clinical Excellence (NICE) in the United Kingdom. Through this analysis, which draws on prior critiques and experiences of NICE, we highlight several critical issues for pharmaceutical priority setting that must be considered in the operation and appraisal of centralized drug review processes. These include the selection of drugs for review, centralized versus decentralized decision-making, receptor capacity at local decision making levels, and public participation. PMID- 16214258 TI - A comparison of individual and social time trade-off values for health states in the general population. AB - This study aimed to compare directly elicited individual time trade-off (TTO) values in a general population sample with the social values derived using the UK EQ-5D index tariff. In the Stockholm County 1998 postal Public Health Survey (n=4950, 20-88 years), the EQ-5D self-classifier, a TTO and a rating scale (RS) question were included (n=2549 for all three questions). The mean TTO (EQ-5D) value was 0.943 (0.890) in the youngest age-group and 0.699 (0.733) in the oldest age-group. The difference between TTO and EQ-5D values was greater in more severe health status groups was. The same equation as for the UK EQ-5D index tariff was estimated for TTO and RS and resulted in significant and consistent coefficients for nearly all dimensions. The coefficients for moderate problems were closer to the EQ-5D index tariff than the coefficients for severe problems. Age was also significant after controlling for the EQ-5D dimensions (p<0.05). The results suggest that individual and social TTO values differ systematically and that the difference is greater the more severe the health status is. The social EQ-5D index tariff may also underestimate the severity in health status at older ages; age appears to correlate with additional health problems not captured by the EQ 5D classification. PMID- 16214259 TI - Interphase mass transfer during chemical oxidation of TCE DNAPL in an aqueous system. AB - The chemical oxidation of trichloroethene dense non-aqueous phase liquid by permanganate was studied in an aqueous system using micro-reaction/extraction vessels in a novel approach. Experiments were conducted at ambient temperature ( approximately 20 degrees C) under static and mixed conditions to evaluate the rate of TCE(DNAPL) dissolution as a function of permanganate concentration. Chemical oxidation by permanganate was shown to increase the rate of TCE(DNAPL) dissolution under static conditions and decrease the rate of dissolution under mixed conditions. The apparent inconsistency in results appears to result from the local deposition of a film at the DNAPL interface composed of manganese oxide solids as discovered through visual observation with the aid of a Goniometer. Data from interfacial deposition experiments suggest that the film formed rapidly and reached maturation within approximately 2 h with little or no growth occurring thereafter. A conceptual model of the reaction and mass transfer processes occurring at the DNAPL interface was proposed based on the experimental results. PMID- 16214260 TI - Possible phosphate interference with copper-silver ionization for Legionella control. PMID- 16214261 TI - Screening for methicillin-resistant Staphylococcus aureus: which anatomical sites to culture? PMID- 16214262 TI - Laundering of hospital staff uniforms at home. AB - Few hospitals now launder staff uniforms. Staff are expected to use their own domestic machines, most of which run with 40 degrees C cycles. However, there is little information on the effectiveness of home laundering. This study demonstrates that domestic washing machines reduce viable counts of Staphylococcus aureus to below detectable levels from an inoculum of 10(8)-10(12) colony-forming units (>or=10(6)-fold reduction), even using low temperature (40 degrees C) programmes. Environmental organisms, predominantly Gram-negative flora, were introduced from the machine itself but were destroyed by tumble drying or ironing. Domestic laundering of uniforms is an acceptable alternative to hospital laundering if combined with tumble drying or ironing. PMID- 16214263 TI - Efficacy of ethanol-based hand rubs. PMID- 16214264 TI - Nicotine has a permissive role on the activation of metabotropic glutamate 5 receptors coexisting with nicotinic receptors on rat hippocampal noradrenergic nerve terminals. AB - The existence of metabotropic glutamate receptors (mGluRs) on hippocampal noradrenergic nerve terminals and their interaction with coexisting nicotinic acetylcholine receptors (nAChRs) were investigated in superfused rat synaptosomes using [(3)H]-noradrenaline ([(3)H]-NA) release as a readout. The selective agonist of group I mGluRs, (S)-3,5-dihydroxyphenylglycine (DHPG), inactive on its own, acquired ability to release [(3)H]-NA when added together with (-)-nicotine. The effect of DHPG was prevented by 2-methyl-6-(phenylethynyl)-pyridine (MPEP), a selective antagonist of mGluR5, but not by 7-(hydroxyimino)cyclopropane[b]chromen 1-carboxylate ethyl ester (CPCCOEt), selective antagonist of mGluR1. The [(3)H] NA release evoked by (-)-nicotine plus DHPG was totally abrogated by the nAChR antagonist mecamylamine. Veratrine mimicked the permissive role of (-)-nicotine on the activation of mGluR5 mediating [(3)H]-NA release. The mGluR5-mediated component of the [(3)H]-NA release provoked by DHPG plus (-)-nicotine was blocked by xestospongin C, a selective antagonist of inositoltrisphosphate (IP(3)) receptors. It can be concluded that (i) release-enhancing mGluRs of subtype 5 exist on hippocampal noradrenergic axon terminals; (ii) activation of mGluR5 to mediate IP(3)-dependent NA release requires activation of depolarizing nAChRs coexisting on the same terminals. PMID- 16214265 TI - Effects of chronic ethanol administration on the activities and relative synthetic rates of myelin and synaptosomal plasma membrane-associated sialidase in the rat brain. AB - In an attempt to understand the possible mechanism of chronic ethanol-induced generation of asialoconjugates in the brain and consequent behavioral abnormalities, we have studied the effects of chronic ethanol feeding to rats on the plasma membrane sialidase status in the various subcellular fractions of the brain. We determined sialidase activity using 3H-monosialoganglioside (3H-GM3), 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid (4-MU-NeuAC) substrates and Amplex Red (Sialidase) kit. We determined the plasma membrane sialidase protein by Western blot using the anti-plasma membrane sialidase. We also determined its relative synthetic rate (RSR) by the 60 min incorporation of intracranially infused [35S]-methionine (50 microCi/100 g) into immunoprecipitable plasma membrane sialidase. Chronic ethanol administration stimulated the sialidase activity in the total brain homogenate as well as the myelin and synaptosomal membrane fractions, respectively, in all the three experimental models. Chronic ethanol also increased the concentration of the rat brain plasma membrane sialidase protein relative to that of glyceraldehyde-3 phosphate dehydrogenase by 2.4-, 1.62- and 1.51-fold in the total brain homogenate, myelin and synaptosomal membrane fractions, respectively. These increases in plasma membrane sialidase activity and its protein content were due to concomitant increases in their relative synthetic rates by 115% (p < 0.01) and 72% (p < 0.01) in the myelin and synaptosomal membrane fractions, respectively. Thus, our studies clearly show that chronic ethanol induced deglycosylation of brain gangliosides is in part, due to specific up-regulation of plasma membrane sialidase in the myelin and synaptosomal membrane fractions of the brain. This increase in plasma membrane sialidase may be responsible for chronic-ethanol induced physiological and neurological impairment in the brain, presumably due to deglycosylation of gangliosides that are essential for crucial cellular and metabolic activities. PMID- 16214266 TI - In pursuit of neural risk factors for weight gain in humans. AB - Obesity is a multifactorial disease associated with an increased risk of type 2 diabetes, coronary artery disease, cancer, and consequently, with a reduced length of life. Metabolic phenotypes of reduced energy expenditure have been associated with weight gain, but their contribution has been estimated to be relatively small. On the other hand, excessive food intake is likely to be the major determinant of positive energy balances and it is underlied by both non conscious (homeostatic) and conscious (perceptual, emotional, and cognitive) phenomena processed in the brain. Functional neuroimaging is a promising tool to investigate these neural substrates in humans, because it provides a measurement of state-dependent brain regional activity, bridging the gap between neural events and behavioral responses. Using this technology, a few studies have provided the first evidence of functional differences between obese and lean individuals in the brain's response to energy intake and investigated the presence of neural risk factors of weight gain. PMID- 16214267 TI - Alzheimer-associated APP+1 transgenic mice: frameshift beta-amyloid precursor protein is secreted in cerebrospinal fluid without inducing neuropathology. AB - Biomarkers present in the cerebrospinal fluid (CSF) of Alzheimer Disease patients could be instrumental in guiding diagnosis and monitoring of progression of the disease. We have previously reported on the secretion of a frameshifted form of amyloid-beta precursor protein, APP+1, into the CSF of Alzheimer patients and controls. APP+1 is secreted efficiently in controls, but during the progression of Alzheimer Disease, its secretion is reduced and APP+1 accumulates in tangle bearing neurons. Here we describe the generation of a transgenic mouse line expressing APP+1 in the brain. These mice do not suffer from overt pathology or neurodegeneration, suggesting that APP+1 is not neurotoxic. To measure APP+1 levels in the CSF, we serially sampled CSF from the cisterna magna in the same mouse over a period of months. Indeed, APP+1 is secreted into the CSF of the transgenic mice, and APP+1 levels are stable over 1 year. This mouse model may guide the study of secretion deficits as found in Alzheimer Disease. PMID- 16214268 TI - A double-inactivated whole virus candidate SARS coronavirus vaccine stimulates neutralising and protective antibody responses. AB - A double-inactivated, candidate whole virus vaccine against severe acute respiratory syndrome associated coronavirus (SARS-CoV) was developed and manufactured at large scale using fermenter cultures of serum protein free Vero cells. A two step inactivation procedure involving sequential formaldehyde and U.V. inactivation was utilised in order to ensure an extremely high safety margin with respect to residual infectivity. The immunogenicity of this double inactivated vaccine was characterised in the mouse model. Mice that were immunised twice with the candidate SARS-CoV vaccine developed high antibody titres against the SARS-CoV spike protein and high levels of neutralising antibodies. The use of the adjuvant Al(OH)3 had only a minor effect on the immunogenicity of the vaccine. In addition, cell mediated immunity as measured by interferon-gamma and interleukin-4 stimulation, was elicited by vaccination. Moreover, the vaccine confers protective immunity as demonstrated by prevention of SARS-CoV replication in the respiratory tract of mice after intranasal challenge with SARS-CoV. Protection of mice was correlated to antibody titre against the SARS-CoV S protein and neutralising antibody titre. PMID- 16214269 TI - The immune response of the Chacma baboon to Bacille Calmette Guerin: development of a primate model for BCG-based vaccine research. AB - Mycobacterium bovis Bacilli Calmette-Guerin (BCG) is used increasingly as an efficient vector for expression of recombinant proteins to induce a strong cell mediated immunity. Here, we tested the immune response of Chacma baboons to the Tokyo and Pasteur strains of BCG in order to obtain base-line information on the response of this primate to BCG. While a humoral immune response to BCG was detected only in some vaccinated baboons, a cellular immune response characterized by a PPD-specific delayed hypersensitivity response and BCG specific IFN-gamma production from PBMC was a consistent finding. These responses were long-lived and could be detected beyond a year after a booster inoculation at 20 weeks. The results thus suggest that the Chacma baboon may be used as a non human primate for the evaluation of recombinant BCG vaccines. PMID- 16214270 TI - Haemolytic activities and adjuvant effect of Bupleurum chinense saponins on the immune responses to ovalbumin in mice. AB - In this study, the haemolytic activities of Bupleurum chinense saponins (BCS) and its adjuvant potentials on the immune responses of ICR mice against ovalbumin (OVA) were evaluated. BCS showed a slight haemolytic effect, with its haemolytic percents being 3.32% and 1.19% at the concentrations of 500 and 250 microg/ml, respectively. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing aluminium hydroxide gel (Alum, 200 microg), QuilA (10 and 20 microg) or BCS (50, 100 or 200 microg) on Days 1 and 15. Two weeks later (Day 28), concanavalin A (Con A)-, lipopolysaccharide (LPS)- and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. BCS significantly enhanced the Con A-, LPS-, and OVA-induced splenocyte proliferation in the OVA-immunized mice especially at a dose of 100 microg (P<0.05 or P<0.001). OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were also significantly enhanced by BCS compared with OVA control group (P<0.01 or P<0.001). Moreover, no significant differences (P>0.05) were observed between enhancing effect of BCS and QuilA on the OVA-specific IgG2b antibody responses to OVA in mice. In conclusion, the results suggest that BCS could be safely used as adjuvant with low or non-haemolytic effect. PMID- 16214271 TI - Evaluation of immunity and protective efficacy of a dengue-3 pre-membrane and envelope DNA vaccine in Aotus nancymae monkeys. AB - A dengue (DEN) virus type 3 DNA vaccine expressing pre-membrane and envelope genes was tested for immunogenicity and protective efficacy in Aotus monkeys. Five of six vaccinated animals demonstrated moderate DEN-specific antibody responses as measured by ELISA and virus neutralization in vitro. By contrast, none of the six control animals developed detectable anti-DEN antibodies. When five vaccinated animals were challenged with live DEN-3 virus and viremia determined by PCR amplification of viral RNA in serum samples, one animal was completely protected and two were partially protected as indicated by a decrease in mean days of viremia. The results demonstrate the ability of the DEN-3 DNA vaccine to elicit a neutralizing antibody response and to partially protect against live virus challenge. These findings support the inclusion of this construct in a tetravalent DNA vaccine. PMID- 16214272 TI - Ultrasound-guided core biopsy for the diagnosis of lumps in the neck: results in 82 patients. AB - Ultrasound-guided core biopsy of nodes (n = 78) and other lumps (n = 4) in the neck allowed a firm histological diagnosis in 76 of 82 patients (93%). Of the remaining six patients, histological examination was equivocal in four and valueless in two. Of 18 patients with a diagnosis of lymphoma only 3 required excision of the node to provide additional information for the planning of treatment. PMID- 16214273 TI - Conservative, non-surgical management of patients presenting with impacted lower third molars: a 5-year study. AB - Our aim was to find out what happened over a period of 5 years to fully or partially impacted mandibular third molars that were left alone. The change in emphasis towards non-intervention in patients with asymptomatic impacted wisdom teeth over the past few years was accompanied by considerable debate as to the eventual outcome of such teeth. Examination of a number of factors including smoking, extent of eruption, depth of periodontal pocket, and history of pericoronitis failed to show any predictive factors that would indicate which teeth would subsequently require removal. However, about one-third of the teeth in this series had to be removed within the 5-year period. Although this does not allow a 'lifetime extrapolation', it blurs the edges of our current thinking about asymptomatic wisdom teeth and certainly suggests that further (possibly longer term) studies need to be completed. It does, however, provide little support for the reintroduction of prophylactic removal of wisdom teeth. PMID- 16214274 TI - Novel biomechanics demonstrate gait dysfunction due to hamstring tightness. AB - BACKGROUND: Children with cerebral palsy and hamstring tightness often demonstrate limited terminal swing knee extension. The conventional clinical measure of popliteal angle describes static hamstring tightness, but is not consistent with dynamic limitation. We hypothesize hamstring tightness, determined via modification of the conventional popliteal angle measure, is directly related to decreased terminal swing knee extension in children with cerebral palsy and normal magnitude knee flexion moments. METHODS: Six patients with cerebral palsy and six normal subjects were evaluated via physical examination and instrumented gait analysis. Physical examination included popliteal angle measures at first hamstring resistance to passive extension (R(1)), and end-range extension (R(2)) with the hip in varying degrees of flexion. Passive R(1) data were used to calculate regression equations to predict R(1) during gait, resulting in a novel measure of Available Knee Extension. Hamstring EMG was also compared. FINDINGS: R(1) during physical examination was significantly correlated with Available Knee Extension at terminal swing (Pearson r = -0.7251, P < 0.0001). Patients walked with significantly decreased velocity (0.959 vs. 1.27 m/s, P = 0.0002) and decreased knee extension at terminal swing (25.6 vs. 2.05 degrees, P < 0.0001), in the presence of normal knee flexion moments (-0.289 vs. -0.306 Nm/kg, P = 0.5009), and significantly decreased power absorption (-0.821 vs. -1.43 W/kg, P < 0.0001). Eleven of 12 patient knees demonstrated negative Available Knee Extension at terminal swing, with markedly limited knee extension. Five of 12 normal knees demonstrated negative Available Knee Extension, but this was near full extension. Hamstring EMG onset times were not significantly different. INTERPRETATION: We believe Available Knee Extension, defined on the basis of clinical measures of first resistance to hamstring stretch, provides a biomechanical link between physical examination findings and dynamic limitations in terminal swing knee extension. PMID- 16214275 TI - The mechanical effect of a pelvic belt in patients with pregnancy-related pelvic pain. AB - BACKGROUND: Many patients with pregnancy-related pelvic girdle pain experience relief of pain when using a pelvic belt, which makes its use a common part of the therapy, but there is no in vivo proof of the mechanical effect of the application of a pelvic belt. METHODS: The influence of a pelvic belt on sacroiliac joint laxity values was tested in 25 subjects with pregnancy-related pelvic girdle pain by means of Doppler imaging of vibrations in prone position with and without the application of a pelvic belt. The belt was adjusted just below the anterior superior iliac spines (high position) and at the level of the pubic symphysis (low position). FINDINGS: Sacroiliac joint laxity values decreased significantly during both applications of a pelvic belt (P<0.001). The application of a pelvic belt in high position decreased sacroiliac joint laxity to a significantly greater degree than the application of a belt in low position (P=0.006). The decrease of laxity significantly correlated with the decrease of the score on the active straight leg raise test (r=0.57 for the low position, P=0.003 and r=0.54 for the high position, P=0.005). INTERPRETATION: Application of a pelvic belt significantly decreases mobility of the sacroiliac joints. The decrease of mobility is larger with the belt positioned just caudal to the anterior superior iliac spines than at the level of the pubic symphysis. The findings are in line with the biomechanical predictions and might be the basis for clinical studies about the use of pelvic belts in pregnancy-related pelvic girdle pain. PMID- 16214276 TI - Assessment of ozone effects on nitrate export from Hubbard Brook Watershed 6. AB - The impact of the air pollution ozone on soil N dynamics and temporal and spatial patterns of streamflow nitrate flux at the Hubbard Brook Experimental Forest Watershed 6 during the 1964-1994 period was assessed using aggregated (one-cell) and spatially explicit (208-cell) versions of the SImple NItrogen Cycle (SINIC) model. Simulated ozone effects included reductions in stomatal conductance and plant N demand. Model uncertainty was evaluated using Monte Carlo simulations. Ambient ozone was estimated to cause an additional 0.042 gN/m2 per year of nitrate export, 12% of the mean annual streamflow nitrate flux. The 95% credible interval of this estimate was 0.002-0.083 gN/m2 per year, or 0.72-27.3% of the annual flux. The large uncertainty in this estimate suggests that it may be difficult to identify ozone effects on nitrate export utilizing long term data from a single site. PMID- 16214277 TI - Removal of harmful cyanobacterial blooms in Taihu Lake using local soils. III. Factors affecting the removal efficiency and an in situ field experiment using chitosan-modified local soils. AB - Effects of ionic strength, pH, organic content, cell concentration, and growth phase on the removal of MA cells using chitosan-modified sepiolite were studied in the laboratory. The MA removal efficiency increased with the increase of salinity for normal clay flocculation. In contrast, for chitosan-modified clays/soils, MA removal efficiency increased with the decrease of salinity. The removal efficiency of chitosan-modified sepiolite was not significantly affected by pH (6-9), but dropped dramatically beyond pH 10. Humic acid had a small negative effect on the removal of MA cells. Cells were removed more effectively by clays around the early senescence growth phase than other growth stages. The removal efficiency increased as the cell concentration increased. In a field enclosure of Taihu Lake, a loading of 0.025 g/L chitosan-modified local soils removed 99% algal cells and no increase of chlorophyll-a was observed during the following one month's monitoring process. PMID- 16214278 TI - Metal bioaccumulation and metallothionein concentrations in larvae of Crassostrea gigas. AB - Larval stages of bivalve molluscs are highly sensitive to pollutants. Oysters from a hatchery from Normandy (English Channel) were induced to spawn, and fertilized eggs were exposed to copper or cadmium for 24 h. Metal accumulation (from 0.125 to 5 microg Cu L(-1) and from 25 to 200 microg Cd L(-1)) and MT concentrations were measured in larvae. Compared to controls, larvae accumulated copper and cadmium with an increase in MT concentrations particularly with cadmium (i.e. 130.96 ng Cu (mg protein)(-1) and 12.69 microg MT (mg protein)(-1) at 1 microg Cu L(-1) versus 23.19 ng Cu (mg protein)(-1) and 8.92 microg MT (mg protein)(-1) in control larvae; 334.3 ng Cd (mg protein)(-1) and 11.70 microg MT (mg protein)(-1) at 200 microg Cd L(-1) versus 0.87 ng Cd (mg protein)(-1) and 4.60 microg MT (mg protein)(-1) in control larvae). Larvae were also obtained from oysters of a clean area (Arcachon Bay) and a polluted zone (Bidassoa estuary) and exposed to copper in the laboratory, their MT concentration was measured as well as biomarkers of oxidative stress. Biomarker responses and sensitivity to copper for the larvae from Arcachon oysters were higher than for those from Bidassoa. PMID- 16214279 TI - Lead and zinc accumulation and tolerance in populations of six wetland plants. AB - Wetland plants such as Typha latifolia and Phragmites australis have been indicated to show a lack of evolution of metal tolerance in metal-contaminated populations. The aim of the present study is to verify whether other common wetland plants such as Alternanthera philoxeroides and Beckmannia syzigachne, also possess the same characteristics. Lead and zinc tolerances in populations of six species collected from contaminated and clean sites were examined by hydroponics. In general, the contaminated populations did not show higher metal tolerance and accumulation than the controls. Similar growth responses and tolerance indices in the same metal treatment solution between contaminated and control populations suggest that metal tolerance in wetland plants are generally not further evolved by contaminated environment. The reasons may be related to the special root anatomy in wetland plants, the alleviated metal toxicity by the reduced rooting conditions and the relatively high innate metal tolerance in some species. PMID- 16214280 TI - The male heart and the female mind: a study in the gendering of antidepressants and cardiovascular drugs in advertisements in Irish medical publication. AB - Stereotypes which suggest that cardiovascular disease and depression are related to gender can have consequences for the mental and physical health outcomes of both men and women. This study examines how these stereotypes may be reinforced by medical publications advertising for cardiovascular and antidepressant medication. A random sample of 61 (with no repeats) advertisements which appeared in Irish medical publications between July 2001 and December 2002 were analysed using both content and semiotic analysis. Results indicate that the meanings created by advertisers for cardiovascular drugs and antidepressants did in fact gender these products. Women were depicted as the predominant users of antidepressants and men as the main users of cardiovascular drugs. The images used identified two stereotyped patients: the 'male' heart patient and the depressed 'female' patient. Furthermore, the imagery and language used to promote the two categories of medication tended to strengthen gendered associations. PMID- 16214281 TI - Team leadership and patient outcomes in US psychiatric treatment settings. AB - Previous studies suggest that psychiatric patients mirror the behaviors of the staff members who treat them, but there is little empirical evidence about how staff dynamics affect patients over time. The goals of this study were to examine associations between: (1) team leader discipline and mutual respect among treatment team members; and (2) mutual respect among team members and improvements in patient quality of life. Two models were tested on data from psychiatric treatment teams within the US Veterans Administration. The first examined associations between the discipline of each team's emergent leader and the level of mutual respect among that team's members. The second model tested associations between mutual respect among staff and changes over time in patients' quality of life. The subjects for model 1 were psychiatric staff members (n=785) whose responses were aggregated for team-level analyses (n=78). Mutual respect was highest in social worker-led teams and lowest in physician-led teams. The subjects for model 2 were 1,638 seriously mentally ill patients in 44 of the units examined in the first model. When mutual respect among staff was greater, patients improved more over time in their satisfaction with the quality of their housing, relations with families, social life, and finances. Together, these analyses imply that mutual respect may improve patient outcomes and that leadership by some disciplines may facilitate such dynamics. In general, leaders may consider learning from other disciplines' strengths to improve their impact. PMID- 16214282 TI - Central organization of androgen-sensitive pathways to the hypothalamic-pituitary adrenal axis: implications for individual differences in responses to homeostatic threat and predisposition to disease. AB - Despite clear evidence of the potency by which sex steroids operate on the hypothalamic-pituitary-adrenal (HPA) axis and genuine sex differences in disorders related to HPA dysfunction, the biological significance of this remains largely ignored. Stress-induced increases in circulating glucocorticoid levels serve to meet the metabolic demands of homeostatic threat head-on. Thus, the nature of the stress-adrenal axis is to protect the organism. As one develops, matures, and ages, still newer and competing physiological and environmental demands are encountered. These changing constraints are also met by shifts in sex steroid release, placing this class of steroids beyond the traditional realm of reproductive function. Here we focus on the dose-related and glucocorticoid interactive nature by which testosterone operates on stress-induced HPA activation. This provides an overview on how to exploit these characteristics towards developing an anatomical framework of testosterone's actions in the brain, and expands upon the idea that centrally projecting arginine vasopressin circuits in the brain act to register and couple testosterone's effects on neuroendocrine and behavioural responses to stress. More generally, the work presented here underscores how a dual adrenal and gonadal systems approach assist in unmasking the bases by which individuals resist or succumb to stress. PMID- 16214283 TI - Fusariotoxin transfer in animal. AB - Mycotoxin fusariotoxins, essentially represented by trichothecenes, zearalenone and fumonisins, are widely scattered in cereals and their products. Human and animals are particularly concerned by toxicity consecutive to oral chronic exposure. Human exposure can be direct via cereals or indirect via products of animals having eaten contaminated feed. As this alimentary risk is considered as a major problem in public health, it is thus of great importance to determine bioavailability, metabolic pathways and distribution of these mycotoxins in animal and human organism. Most studies indicate that fusariotoxins can be rapidly absorbed in the small intestine but the mechanisms involved remain unclear. Except NIV, fusariotoxins can be partly metabolised into more hydrophilic molecules in digestive tract or liver. Fumonisins present different behaviour as they seem very few and slowly absorbed and metabolised. The main part of absorbed fusariotoxins shows a rapid elimination within 24h after ingestion, followed by a slower excretion of small amounts. However, traces of fusariotoxins or their derivates can be found in animal products. This manuscript, reviewing literature published on fusariotoxin transfer, highlights that too little data are available to correctly appreciate fusariotoxin transfer in organism. Further studies focusing on mechanisms involved in the transfer are needed before clarifying risk assessment for human health. PMID- 16214284 TI - Microtensile bond strength of a filled vs unfilled adhesive to dentin using self etch and total-etch technique. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of a filled adhesive (One-Step Plus; Bisco) versus an unfilled adhesive (One-Step; Bisco) on the microtensile bond strength (muTBS) to dentin using total-etch (Uni-etch; Bisco) and self-etch (Tyrian SPE; Bisco) techniques. METHODS: Twenty extracted human third molars were ground flat to expose occlusal dentin. After the dentin surfaces were polished with 600-grit SiC paper, the teeth were randomly assigned to four groups according to the bonding agent and technique being used. Dentin surfaces were bonded with One-Step Plus+total-etch; One-Step Plus+self-etch; One Step+total-etch and One-Step+self-etch. Composite buildups were performed with Clearfil AP-X (Kuraray Medical). Following storage in distilled water at 37 degrees C for 24 h, the bonded specimens were serially sectioned into 0.7 mm thick slabs and then trimmed to hour-glass shapes with a 1 mm2 cross-sectional area (n=20). Microtensile bond strengths were determined using the EZ-test (Shimadzu) at a cross-head speed of 1 mm/min. Data were analyzed using two-way ANOVA and Tukey's post hoc test. RESULTS: There were no significant differences in the muTBS between One-Step Plus and One-Step adhesives when they were used with the total-etch and self-etch techniques (p>0.05). However with the total etch technique both adhesives yielded significantly higher bond strength values than the self-etch technique (p<0.001). CONCLUSION: The filled adhesive One-Step Plus did not show any beneficial effect than the unfilled adhesive One-Step on the muTBS to dentin with total-etch and self-etch techniques. Irrespective from the adhesive type, self-etch technique revealed lower bond strengths than the total-etch technique. PMID- 16214285 TI - Analysis of an administrative database of half a million restorations over 11 years. AB - AIM: This paper describes the analytical approach used to extract empirical distributions of the interval from the date of placement of a direct restoration to the date when the next intervention took place on the same tooth, that is, an estimate of restoration survival. METHODS: Data, based on the complete attendance and treatment history, over the eleven years from 1991 to 2001, of a statistically representative sample of 82,537 General Dental Services' patients in England and Wales, all of whom received at least one directly placed restoration during the observation period, have been analysed. The patients on the database received a total of 719,009 courses of treatment, and there were 503,965 occasions when a tooth was directly restored. The method of analysis involved, first, the estimation of the probability that the patient will eventually return, given an interval without attending, by analysing the observed patterns of re-attendance. This estimated probability of re-attendance was then used to modify the standard Kaplan-Meier procedure to produce realistic estimates of the hazard of re-intervention. RESULTS: The results demonstrate that the newly developed methodology has produced robust estimates of the distribution of survival intervals to next intervention on the same tooth. Regarding attendance patterns, re-attendance probability varies with length of interval since last attendance. There is also a strong association with patient age. The older the patient, the more likely it is that a gap in attendance is indicative that the patient will never return. The detailed cumulative survival curve over eleven years has been plotted and forty-seven per cent of restorations, overall, survived without re-intervention for at least ten years. The entire analysis was then successfully replicated on a second, independently selected, sample of attendance and treatment records, confirming the ten-year survival estimate. CONCLUSIONS: This newly developed methodology has produced robust estimates of the distribution of survival intervals to next intervention on the same tooth. Patient re-attendance probability varies with length of interval since last attendance and with patient age. The ten-year overall survival rate to next intervention on the same tooth for direct restorations placed within the GDS in England and Wales is circa 47%. PMID- 16214286 TI - Modes of delivery of first and second twins as a function of their presentation. Study of 614 consecutive patients from 1992 to 2000. AB - OBJECTIVES: To study modes of delivery and neonatal morbidity of twins as a function of their presentation. STUDY DESIGN: The study related to 614 consecutive patients who gave birth to twins in the maternity ward of the Hopital Robert Debre from 1992 to 2000. Group A (n=529) included patients who gave birth after 33 weeks of gestation and group B (n=85) before 33 weeks. The parameters studied were the mode of delivery, the need for intubation at birth, 5-min Apgar score <9, transfer to intensive care, death in the neonatal period. The chi square test was used for statistical analysis. RESULTS: In group A, significantly more cesarean sections were performed for breech-breech (22.6%) and breech-vertex (16%) presentations than for vertex-vertex (10.3%) and vertex-breech (4.6%) presentations. In group B, there was no significant difference in the rate of vaginal delivery. Second vertex twins of group A had significantly higher frequencies of intubation (3%) at birth and transfer to intensive care (3%) than the vertex first twins born by vaginal delivery (p=0.01). These percentages were not significantly different from those observed for the second twins born by planned Cesarean section before the start of labor (2.8 and 5.6%, respectively). In group B, neonatal parameters did not differ significantly with the type of presentation at delivery. CONCLUSIONS: The type of presentation should not influence the choice of mode of delivery of twin pregnancies, whatever the gestational age. PMID- 16214287 TI - Flexibility in starfish behavior by multi-layered mechanism of self-organization. AB - Understanding animal behavior as a product of natural selection sometimes result in an underestimation of the animal's adaptability: lower animals with poor mental capabilities are usually considered to simply exhibit innate behavioral patterns. Self-organized behavior may exhibit both stability of certain behavioral patterns and flexibility in adopting those patterns. Thus, the self organization processes of starfish arm and tube feet movements are investigated, by observing obstacle avoidance behavior and tube feet of moving starfish. As starfish have no central nervous systems, their behaviors are the result of certain self-organization processes. Starfish have hierarchically constructed motor organs consisting of arms and tube feet. The collective behavior of the tube feet does not function only as simple fluctuations in the arms' coordination. As a result, starfish seem to exhibit more versatile behavioral changes than expected from the original model of a self-organized behavior. PMID- 16214288 TI - The remediation performance of a specific electrokinetics integrated with zero valent metals for perchloroethylene contaminated soils. AB - This research was conducted to evaluate an integrated technique, combination of the electrokinetics (EK) and zero-valent metal (ZVM), for remediation of the perchloroethylene (PCE) contaminated soils. Various experimental conditions were controlled such as different voltage gradients, the position of ZVM, and ZVM species. The appropriate operational parameters are concluded as follows: (1) 0.01 M sodium carbonate serves as the working solution; (2) the voltage gradient is controlled at 1.0 V/cm; (3) ZVM wall is settled close to the anode. Based on the above operation conditions, the pH value of working solution can maintain at neutral range for avoiding the soil acidification. Neutral pH also causes the system to stay at a stable status of electricity consumption. The removal efficiency reaches 99% and 90% for decontaminating the PCE in the pore-water and the soil, respectively, after a 10-day treatment. The zero-valent zinc performs better PCE degradation than zero-valent iron. Moreover, the soils treated by EK+ZVM still possess their original properties. PMID- 16214289 TI - Regulation of the expression of GABAA receptor subunits by an antiepileptic drug QYS. AB - It has been reported that the antiepileptic drug qingyangshenylycosides (QYS) modulated the function of GABAergic system. However, little is known about the effects of QYS on the gene expression of GABA receptors in the central nervous system (CNS). In the present study, we examined the effects of QYS on the expression of GABAA receptor subunits in different regions of the mouse brain. The results showed that treatment of QYS significantly increased the expressions of Gabra1, Gabra2 and Gabr4 and decreased the expression of Gabrg2 in inferior colliculus. Moreover, Gabrb2 expression was up-regulated and Gabra5 was down regulated in hippocampus, while the expressions of Gabra1 and Gabrb2 were induced in cortex after QYS treatment. These data indicated that QYS had different effects on the expression of GABAA receptor subunits in different brain regions. These results may help to reveal the molecular mechanism of anticonvulsant action of QYS. PMID- 16214290 TI - The UBQLN1 polymorphism, UBQ-8i, at 9q22 is not associated with Alzheimer's disease with onset before 70 years. AB - An intronic polymorphism affecting alternative splicing of exon 8 of ubiquilin 1 (UBQLN1), that is located at a well established Alzheimer's disease (AD) locus on chromosome 9q22, was recently associated with increased risk for late-onset AD. We analyzed this polymorphism in two independent AD samples consisting of patients with an onset age 70 years or less, but did not observe statistically significant association. Our study does not support a major role for this UBQLN1 polymorphism in AD patients with an earlier onset of disease. PMID- 16214291 TI - Differential expression of selectins by mouse brain capillary endothelial cells in vitro in response to distinct inflammatory stimuli. AB - Increased lymphocyte trafficking across blood-brain barrier (BBB) is a prominent and early event in inflammatory and immune-mediated CNS diseases. The adhesion molecules that control the entry of leukocytes into the brain have not been fully elucidated. Although the role of ICAM-1 and VCAM-1 has been well documented, the expression and role of selectins is still a matter of controversy. In a mouse syngenic in vitro BBB model, highly relevant for examining immunological events, mouse brain capillary endothelial cells (MBCECs) do not express selectins. Treatment of MBCECs with LPS, induced E- and P-selectin expression, whereas TNF alpha or IFN-gamma treatments did not. Finally, P-selectin but not E-selectin expression was induced in IL-1beta treated MBCECs. Thus, our study suggests that diverse inflammatory stimuli could differentially regulate selectin expression at the BBB. PMID- 16214292 TI - Motor cortex stimulation for refractory neuropathic pain: four year outcome and predictors of efficacy. AB - Thirty-one patients with medically refractory neuropathic pain were included in a prospective evaluation of motor cortex stimulation. The long-term outcome was evaluated using five variables: (a) rate (percentage) of pain relief, (b) pain scores as assessed on VAS, (c) postoperative decrease in VAS scores, (d) reduction in analgesic drug intake, (e) a dichotomic (yes/no) response to the question whether the patient would accept, under similar circumstances, to be operated on again. Pain relief was rated as excellent (>70 % pain relief) in 10 % of cases, good (40-69 %) in 42 %, poor (10-39 %) in 35 % and negligible (0-9 %) in 13 %. Intake of analgesic drugs was decreased in 52 % of patients and unchanged in 45 % (unavailable data in 3 %), with complete withdrawal of analgesic drugs in 36 % of patients. Twenty-one patients (70 %) declared themselves favourable to re-intervention if the same beneficial outcome could be guaranteed. Neither preoperative motor status, pain characteristics, type or localisation of lesions, quantitative sensory testing, Somatosensory Evoked Potentials, nor the interval between pain and surgery were found to predict the efficacy of MCS. The level of pain relief, as evaluated in the first month following implantation was a strong predictor of long-term relief (regression analysis, R=0.744; p<0.0001). These results confirm that MCS can be a satisfactory and durable alternative to medical treatments in patients with refractory pain, and suggest that the efficacy of MCS may be predicted in the first month of therapy. PMID- 16214293 TI - Thermodynamic basis for antibody binding to Z-DNA: comparison of a monoclonal antibody and its recombinant derivatives. AB - Antibody engineering represents a promising area in biotechnology. Recombinant antibodies can be easily manipulated generating new ligand and effector activities that can be used as prototype magic bullets. On the other hand, an extensive knowledge of recombinant antibody binding and stability features are essential for an efficient substitution. In this study, we compared the stability and protein binding properties of two recombinant antibody fragments with their parental monoclonal antibody. The recombinant fragments were a monomeric scFv and a dimeric one, harboring human IgG1 CH2-CH3 domains. We have used fluorescence titration quenching to determine the thermodynamics of the interaction between an anti-Z-DNA monoclonal antibody and its recombinant antibody fragments with Z-DNA. All the antibody fragments seemed to bind DNA similarly, in peculiar two-affinity states. Enthalpy-entropy compensation was observed for both affinity states, but a marked entropy difference was observed for the monomeric scFv antibody fragment, mainly for the high affinity binding. In addition, we compared the stability of the dimeric antibody fragment and found differences favoring the monoclonal antibody. These differences seem to derive from the heterologous expression system used. PMID- 16214294 TI - Tamoxifen does not improve overall survival in people with advanced-stage hepatocellular carcinoma. PMID- 16214295 TI - Elevated blood pressure may cause volume reductions of temporal areas and induce psychotic episodes. PMID- 16214296 TI - Correlation of probiotic Lactobacillus salivarius growth phase with its cell wall associated proteome. AB - Lactobacillus salivarius subsp. salivarius UCC118 is a probiotic bacterium that was originally isolated from human intestinal tissues and was subsequently shown in a pilot study to alleviate symptoms associated with mild-moderate Crohn's disease. Strain UCC118 can adhere to animal and human intestinal tissue, and to both healthy and inflamed ulcerative colitis mucosa, irrespective of location in the gut. In this study, an enzymatic technique has been combined with proteomic analysis to correlate bacterial growth phase with the presence of factors present in the cell wall of the bacterium. Using PAGE electrophoresis, it was determined that progression from lag to log to stationary growth phases in vitro correlated with increasing prominence of an 84kD protein associated with in vitro adherence ability. Isolated proteins from the 84kD band region were further separated by two-dimensional electrophoresis, resolving this band into 20 individual protein spots at differing isoelectric points. The protein moieties were excised, trypsin digested and subjected to tandem mass spectrometry. The observed proteins are analogous to those reported to be associated with the Listeria monocytogenes cell wall proteome, and include DnaK, Ef-Ts and pyruvate kinase. These data suggest that at least some of the beneficial attributes of probiotic lactobacilli, and in particular this strain, may be due to nonpathogenic mimicry of pathogens and potentially be mediated through a form of attenuated virulence. PMID- 16214297 TI - Isolation and characterisation of a partial peptide synthetase gene from Trichoderma asperellum. AB - Many species of Trichoderma have attracted interest as agents for the biological control of soil borne fungal pathogens of a range of crop plants. Research on the biochemical mechanisms associated with this application has focused on the ability of these fungi to produce enzymes which lyse fungal cell walls, and antifungal antibiotics. An important group of the latter are the non-ribosomal peptides called peptaibols. In this study Trichoderma asperellum, a strain used in biological control in Malaysia, was found to produce the peptaibol, trichotoxin. This type of peptide molecule is synthesised by a peptide synthetase (PES) enzyme template encoded by a peptide synthetase (pes) gene. Using nucleotide sequences amplified from adenylation (A-) domains as probes, to hybridise against a lambda FIXII genomic library from T. asperellum, 25 clones were recovered. These were subsequently identified as representative of four groups based on their encoding properties for specific amino acid incorporation modules in a PES. This was based on analysis of their amino acid sequences which showed up to 86% identity to other PESs including TEX 1. PMID- 16214299 TI - Plasma estrone sulphate (E1S) and estradiol-17beta (E2beta) profiles during pregnancy and their relationship with the relaxation of sacrosciatic ligament, and prediction of calving time in Holstein-Friesian cattle. AB - The objectives of this study were to investigate the plasma E(1)S and E(2)beta profiles during pregnancy and their relationship with the relaxation of sacrosciatic ligament in Holstein-Friesian cattle (n=37) and then to predict the calving time on the basis of E(1)S and E(2)beta profiles and relaxation of the ligament. Blood samples were collected at 4 weeks intervals from days 100 to 190, at 2 weeks intervals from days 190 to 250, every week from days 250 to 270 and thereafter every day from day 270 of gestation until the day after calving. The relaxation in the ligament was measured by using two scales as a distance at a schedule similar to blood sampling plus 5 days postpartum. One scale was kept firm exactly parallel to the ligament between the sacrum and the tuber ischii and other scale was erected perpendicularly to the first scale with the bottom just touching the ligament and the depth was measured in the second scale from the point where it touched the ligament to the point where it touched the first scale. Plasma samples were analyzed for E(1)S and E(2)beta by enzyme immunoassay. E(1)S concentration was low at day 100 (0.8+/-0.3 ng/ml), then increased progressively and drastically to reach the level of 28.4+/-3.6 ng/ml on the day before calving and declined significantly (p<0.05) at 9.5+/-3.1 ng/ml within 1 day postpartum. There was a gradual increase in concentration of E(2)beta from day 100 of gestation (0.1+/-0 ng/ml) until day 4 prepartum (0.6+/-0 ng/ml). Thereafter, it increased drastically and reached the peak level of 1.0+/-0.1 ng/ml (p<0.05) on the day before calving and declined markedly at 0.4+/-0.1 ng/ml within 1 day postpartum (p<0.05). Corresponding to E(1)S and E(2)beta concentrations, a gradual increase in the relaxation of the ligament was observed from day 100 of gestation (8+/-1mm) until day 2 prepartum (24+/-2mm). Thereafter, it showed a significant increase (p<0.05) within 1 day before calving (31+/-2mm) and almost no difference between day 1 prepartum and day 1 postpartum. A marked decrease (p<0.05) was observed thereafter until day 3 postpartum (10+/-2mm) and no significant change between days 3 and 4 as well as 4 and 5 postpartum. The increment of E(2)beta by >or=0.20 ng/ml from the preceding day concentration was 85.2% accurate for predicting calving within 24h in many of the cows (23 of 37) in the herd. The increment in ligament relaxation measurement by >or=5mm from the preceding day measurement was the most efficacious to predict calving within 24h with the highest accuracy (93.9%) in high proportions of cows (31 of 37) in the herd. In conclusion, plasma E(1)S and E(2)beta concentrations and relaxation of sacrosciatic ligament increased gradually as gestation advanced and reached the peak level on the day before calving. The relaxation in the ligament corresponded well to plasma E(2)beta concentrations. Prediction of calving was possible by E(2)beta profile and relaxation in the ligament but not by E(1)S profile. The increment in ligament measurement by >or=5mm from the preceding day measurement was the most useful and accurate in predicting calving within 24h. It is economical and easily applicable in the field condition. PMID- 16214298 TI - A cascade analysis of the interaction of mercury and coproporphyrinogen oxidase (CPOX) polymorphism on the heme biosynthetic pathway and porphyrin production. AB - Mercury (Hg) exposure in various forms remains a persistent public health concern in many parts of the world. In previous studies, we have described a biomarker of mercury exposure characterized by increased urinary concentrations of specific porphyrins, pentacarboxyporphyrin (5-CP) and coproporphyrin (4-CP), and the atypical keto-isocoproporphyrin (KICP), based on selective interference with the fifth (uroporphyrinogen decarboxylase, UROD) and sixth (coproporphyrinogen oxidase, CPOX) enzymes of the heme biosynthetic pathway. Whereas this response occurs in a predictable manner among approximately 85% of subjects with Hg exposure, an atypical porphyrinogenic response (APR) has been observed in approximately 15% of Hg-exposed persons, in which the three porphyrins that are affected by Hg, i.e., 5-CP, 4-CP and, KICP, are excreted in substantial excess of that predicted on the basis of Hg exposure alone. This APR has been attributed to a specific polymorphism in exon 4 of the CPOX gene (CPOX4). In the present study, we sought to further confirm the hypothesis that the observed changes in porphyrin excretion patterns might serve as a biomarker of Hg exposure and potential toxicity by statistically modeling the cascading effects on porphyrin concentrations within the heme biosynthetic pathway of Hg exposure and CPOX4 polymorphism in a human population with long-term occupational exposure to elemental mercury. Our results are highly consistent with this hypothesis. After controlling for precursor porphyrin concentrations, we demonstrated that 5-CP and 4-CP are independently associated with Hg concentration, while KICP is associated only with the CPOX4. An unpredicted association of Hg with heptacarboxyporphyrin (7-CP) may indicate a previously unidentified point of mercury inhibition of UROD. These findings lend further support to the proposed utility of urinary porphyrin changes as a biomarker of exposure and potential toxicity in subjects with mercury exposure. Additionally, these findings demonstrate the successful application of a computational model for characterizing complex metabolic responses and interactions associated with both toxicant exposure and genetic variation in human subjects. PMID- 16214300 TI - PAMAM dendrimers and model membranes: differential scanning calorimetry studies. AB - Dendrimers attract much attention as potential drug and gene carriers for intracellular delivery. From this point of view, it is crucial to extend our knowledge about their interactions with membranes. The influence of polyamidoamine (PAMAM) dendrimers on the thermotropic behavior of DPPC multilamellar vesicles and DMPC small unilamellar vesicles was examined by differential scanning calorimetry. We used three types of PAMAM dendrimers to determine how a dendrimer structure determines interactions with liposomes. We show that the strength of interactions depends on both the dendrimers' structure and degree of hydrophobicity. A model for the interaction of each type of dendrimer with liposomes was proposed. PMID- 16214301 TI - Treatment of Trypanosoma cruzi-infected mice with propolis promotes changes in the immune response. AB - Ethanol extract of Bulgarian propolis (Et-Blg) was administered by oral route in doses ranging from 25 to 100mg/kg body weight in experimental Trypanosoma cruzi infected Swiss mice. Treatment with 50mg Et-Blg/kg body weight/day led to a decrease in parasitemia and showed no hepatic or renal toxic effect. Treatment with Et-Blg led to a decrease in the spleen mass and modulated the initial inflammatory reaction as demonstrated by analysis of the leukocyte profile in peripheral blood, quantification of T cells subsets, and phenotypic markers in the spleen. Preferential expansion of CD8(+) and partial inhibition in the increase of CD4(+)CD69(+) and CD8(+)CD69(+) in CD4(+)CD44(+) and CD8(+)CD44(+) and in the decrease of CD8(+)CD62L in Trypanosoma cruzi-infected mice were also observed. Taken together, our data indicate that treatment of Trypanosoma cruzi infected mice with Et-Blg interferes with the basic properties of immune cells. PMID- 16214302 TI - Use of medicinal plants for the treatment of oral diseases in Burkina Faso. AB - This paper presents the findings of an investigation into the treatment of oral diseases with medicinal plants in the Kadiogo province, Burkina Faso. Although the region is mainly urban, it appears that traditional healers who live there, and the general population, continue to rely on plant products when dealing with a broad range of oral health concerns. Sixty-two relevant species belonging to 29 families were identified, and each was documented with regard to its local name, part used, indication, mode of administration, and the collection and storage procedure used (when those data were available). A number of ideas for research topics emerged from this work, some of which promise to help selection and prescription of improved traditional remedies for oral diseases at the primary health care level in Burkina Faso. PMID- 16214303 TI - Radiological assessment of bone segments for transplantation: experience at Rizzoli Orthopedic Institute. AB - AIM: To analyse results obtained from radiological assessment of skeletal segments stored in the musculoskeletal tissue bank (MTB) at Rizzoli Orthopaedic Institute. MATERIALS AND METHODS: Between January 1997 and June 2003, 891 bone segments underwent radiographic examination in two views. Two hundred and thirteen of these segments were examined by a radiologist between July 2002 and June 2003. Diagnostic evaluation was aimed at recognising relevant degenerative, traumatic and focal lesions. Focal lesions underwent histological tests. RESULTS: Twenty-two lytic lesions were found in 12 segments. Ten of these were studied between July 2002 and June 2003 and two in the period before specialist radiological assessment. In the latter cases the lesions were identified by the orthopedic specialist who had examined the X-rays before planning surgery. Histological tests showed that the bone tissue was normal or involved by degenerative phenomena. CONCLUSIONS: We think donor screening should include radiological assessment of bone segments, performed according to standard parameters by a radiologist to identify bone lesions that may jeopardise the successful outcome of surgery. PMID- 16214304 TI - Home cage observations following acute and repeated IV cocaine in intact and gonadectomized rats. AB - The purpose of the present experiment was to examine the effects of acute and repeated intravenous (IV) cocaine on rat behavior in the home cage environment. An observational sampling method was used. Pair-housed, male, female, castrated (CAST), and ovariectomized (OVX) rats were administered daily IV cocaine injections (3.0 mg/kg/injection) in the home cage for 13 consecutive days, and observations occurred after the 1st and 13th injections. The incidence, i.e., occurrence or nonoccurrence of a behavior, was recorded according to a behavioral profile comprised of 11 behaviors. Data were analyzed as locomotor composite and orofacial composite scores. Behaviors not amenable for combination into a composite incidence score were evaluated independently (e.g., still behavior). Females exhibited more locomotor incidence scores than males following acute injection and more still behavior after repeated cocaine administration. Females exhibited more locomotor activity than OVX rats following acute, but not repeated, cocaine injection. There were no differences between the male and CAST rats on days 1 or 13. CAST rats exhibited more still behavior than OVX following only acute cocaine administration. This study indicates that IV cocaine-induced sex differences and the effects of gonadectomy can be measured in the home cage, and furthermore, describes a simple method to screen changes in cocaine-induced locomotor behaviors in the absence of automated equipment. PMID- 16214305 TI - The effect of RGS12 on PDGFbeta receptor signalling to p42/p44 mitogen activated protein kinase in mammalian cells. AB - We have previously shown that the PDGFbeta receptor uses a classical GPCR mediated pathway in order to induce efficient activation of p42/p44 MAPK in response to PDGF. We therefore, considered the possibility that GTPase accelerating proteins (RGS proteins), which regulate GPCR signalling, modulate PDGFbeta receptor-mediated signal transmission. Several lines of evidence were obtained to support functional interaction between the PDGFbeta receptor and RGS12 in HEK 293 and airway smooth muscle cells. Firstly, the over-expression of the RGS12 PDZ/PTB domain N-terminus or RGS12 PTB domain reduced the PDGF-induced activation of p42/p44 MAPK. Secondly, the RGS12 PDZ/PTB domain N-terminus and RGS12 PDZ domain can form a complex with the PDGFbeta receptor. Therefore, the results presented here provide the first evidence to support the concept that the PDZ/PTB domain N-terminus and/or the PTB domain of RGS12 may modulate PDGFbeta receptor signalling. In airway smooth muscle cells, over-expressed recombinant RGS12 and the isolated PDZ/PTB domain N-terminus co-localised with PDGFbeta receptor in cytoplasmic vesicles. To provide additional evidence for a role of the PDZ/PTB domain N-terminus, we used RGS14. RGS14 has the same C-terminal domain architecture of an RGS box, tandem Ras-binding domains (RBDs) and GoLoco motif as RGS12, but lacks the PDZ/PTB domain N-terminus. In this regard, RGS14 exhibited a different sub-cellular distribution compared with RGS12, being diffusely distributed in ASM cells. These findings suggest that RGS12 via its PDZ/PTB domain N-terminus may regulate trafficking of the PDGFbeta receptor in ASM cells. PMID- 16214306 TI - The novel antiepileptic drug lacosamide blocks behavioral and brain metabolic manifestations of seizure activity in the 6 Hz psychomotor seizure model. AB - Brain metabolic activation after 6 Hz electrical stimulation (32 mA, 3s stimulus duration) was assessed by autoradiographic analysis of 14C-2-deoxyglucose (2-DG) uptake. In addition, effects of the new antiepileptic drug lacosamide were examined on the stimulation-induced metabolic activation. The 6 Hz stimulation via corneal electrodes induced a robust increase 2-DG uptake in cerebral cortical regions, lateral amygdala, and the caudate-putamen. Many other brain regions were not affected by the stimulation, including the hippocampal formation, medial nuclei of the amygdala, thalamus, and hypothalamus. Lacosamide (20 mg/kg) injected i.p. 30 min before application of electrical stimulation antagonized completely the seizure-induced brain metabolic activation but did not affect basal 2-DG uptake. The data provide evidence that lacosamide antagonizes the neural activation induced by an electrical seizure stimulus, without suppressing normal brain metabolic activity. PMID- 16214307 TI - Influence of hydrophobe on the release behavior of vinyl acetate miniemulsion polymerization. AB - Poly(vinyl acetate) (PVAc) nanoparticles containing antibiotic have been prepared by miniemulsion polymerization. To compare the effect of hydrophobe types, hexadecane and poly(vinyl acetate) were used as hydrophobe. The particle characteristics as the manufacturing condition were examined by particle size analyzer. As a result, the diameter of PVAc latexes was adjusted between 80 and 260 nm by homogenization conditions and amounts of surfactant. Also, the miniemulsion by using hexadecane showed the more long shelf stability and led to the more small particle size after polymerization, as compared with the case of using poly(vinyl acetate). This indicated that the use of poly(vinyl acetate) as a hydrophobe could not make the stable emulsion, but it could avoid volatile organic chemical problems in the final product. From the release profile of drug through UV spectra, the drug release was very slow and it could be seen that the release of drug encapsulated with PVAc was occurred with the polymer degradation. PMID- 16214308 TI - Effect of surfactants on the dissociation constants of ascorbic and maleic acids. AB - The dissociation equilibria of ascorbic and maleic acids have been studied in certain cationic, anionic and non-ionic micellar media and their pK(a) values have been evaluated by the potentiometric, conductometric and spectrophotometric techniques. These pK(a) values have been found to shift in micellar media as compared to those in pure water. The differences in the values have been attributed to the solvent properties of the interfacial and bulk phases involving contribution from the micellar surface potential in the case of charged micelles. The values of the limiting molar conductance of the acids have been determined in the various surfactants and were found to be different in different surfactants. In case of the cationic surfactants the limiting molar conductance was found to increase with increase in surfactant concentration. PMID- 16214309 TI - Back muscle activation pattern and spectrum in defined load situations. AB - For the prevention and rehabilitation of low back pain the understanding muscle function in the low back region is essential. Important aspects of function include the muscle activation patterns and muscle fatigue. In the low back region m. erector spinae plays a critical role. The different parts of this muscle complex differ considerably in function. Following the concept of Bergmark [A. Bergmark, Stability of the lumbar spine, Acta Orthop. Scand. 60 (1989) 1-54] trunk muscles can be divided in two subsystems, the global-mobilizing-system and the local-stabilizing-system. At present the multifidus muscles are assigned to the local whereas the longissimus and iliocostalis muscles are assigned to the global system. From results of the four parts of our investigation, it can be shown that essential information about muscle function can be obtained by spectral and intra- as well as inter-muscular surface EMG parameters. Obtained data demonstrated functional transitions between the local and global system of trunk muscles for the multifidus muscle. PMID- 16214310 TI - [From child accident prevention at home to safety promotion: the commitment of the international francophone network]. PMID- 16214311 TI - [Use of transesophageal pacing for documentation of older children with accessory pathway]. AB - INTRODUCTION: In case of an accessory pathway, children are exposed to severe cardiac events including sudden death. Radiofrequency ablation is a standardized procedure, which can be applied to a significant number of children although complications can still potentially occur. In this context, transesophageal evaluation of the accessory pathway evaluation can be discussed. MATERIALS AND METHODS: Among 140 procedures performed in 19 years, 70 were done for accessory pathway evaluation. The preexcitation was overt in 59 children older than 5 years, which form the basis in this study. RESULTS: Anterograde refractory period was determined in 88% cases and was found<220 ms in 12 cases justifying an ablation procedure. Conversely, in case of a long refractory period (>250 ms), the ablation procedure was not performed in 8 asymptomatic cases and was postponed in 11/20 mildly symptomatic children. Transesophageal electrophysiologic study seems legitimate in asymptomatic or mildly symptomatic children. CONCLUSION: This technique is probably less useful in case of an overt preexcitation and recurrent reciprocating tachycardia requiring long-term antiarrythmic treatment. In this case, endocavitary electrophysiological study eventually followed by an ablation procedure seems the best option. PMID- 16214312 TI - Practical method for choosing diluent that ensures the best temperature uniformity in the case of pharmaceutical microwave vacuum drying of a heat sensitive product. AB - Microwave vacuum drying is getting more and more popular thanks to its known advantageous and unique features, but its non-uniform electric field can cause nonhomogeneous temperature distribution in the workload. The origin and effect of a generated hot-spot is influenced by the electromagnetic and thermodynamic features of the microwave system and the workload. In the case of single/one pot technology, the geometry and the construction of the microwave cavity is primarily designed for high-shear granulation. As for the workload, its composition has first-order effect on the electric field pattern. The aim of our study is to present a rational decision procedure based on basic practical experiments and the elaborated '3D layered thermography' technique to make it possible to choose the most suitable diluent to formulate a heat sensitive active pharmaceutical ingredient given its stability due to temperature distribution within the workload. Comparing two commonly used diluents, namely microcrystalline cellulose and corn starch, it was found that in the case of different actives with different acceptable temperature limits different diluents are recommended. Drying of a composition consists of an active ingredient characterized by a temperature limit of 70 degrees C. Using corn starch is safer when the workload is less endangered than when using microcrystalline cellulose. Above this temperature limit microcrystalline cellulose becomes beneficial. PMID- 16214313 TI - In vivo human Cartilage oligomeric matrix protein (COMP) promoter activity. AB - Cartilage oligomeric matrix protein (COMP) is a large extracellular matrix protein whose function is unknown. Mutations in COMP cause pseudoachondroplasia and multiple epiphyseal dysplasia, two skeletal dysplasias which are associated with intracellular retention of COMP in chondrocytes. In contrast, COMP null mice are normal suggesting gene redundancy or that the detrimental effect is associated with mutant COMP rather than the absence of functional COMP. To define the elements that regulate COMP transcription and tissue-specificity, we have evaluated the human COMP promoter driving fusion gene expression in vitro and in vivo. COMP promoter activity is higher in rat chondrosarcoma cells (RCS) than in a fibroblast cell line. In RCS cells, expression of a reporter gene containing 1.7 kb of the human COMP promoter was three-fold higher than all shorter COMP promoter constructs. In transgenic mice, 1.7 kb of the human COMP promoter is active early in development in the limbs, spine, and eye. As development progresses, promoter activity diminishes in the eye and migrates from the center to the ends of the long bones. On the other hand, while 375 bp of the human COMP promoter is sufficient for proper tissue-specific expression, levels are less than those found with the 1.7-COMP promoter. The expression pattern of both promoters recapitulates endogenous cartilage COMP expression in mice. Our findings indicate that the elements required for chondrocyte-specific expression lie within 375 bp of the translational start site, while DNA enhancer elements are located between 1.0 to 1.7 kb. PMID- 16214314 TI - The hypereosinophilic syndromes: still more heterogeneity. AB - In 1968, the term hypereosinophilic syndromes (HES) was coined to refer to a spectrum of eosinophil-associated diseases presumed to be caused by an underlying immunological pathology. In the 1990s, the identification of an HES subset with T lymphocyte clonality and production of cytokines, particularly IL-5, validated this concept. Then, in 2002, imatinib mesylate, which was introduced for the treatment of chronic myelogenous leukemia, effectively controlled another subgroup of HES patients. Imatinib's target is a novel constitutively-active kinase. Most imatinib-responsive HES patients show an increased number of bone marrow mast cells and elevated serum tryptase; mast cells, lymphocytes and neutrophils express the novel kinase. This new information critically modifies our view of HES and indicates that several cell lines are altered and likely to contribute to HES pathophysiology. PMID- 16214315 TI - Rethinking genetic models of asthma: the role of environmental modifiers. AB - Asthma is a common, chronic disease with a complex etiology. To date, more than 35 genes have been associated with asthma or related phenotypes in multiple populations, but none of them has been shown to contribute to risk in all populations studied. We suggest that genetic susceptibility is both context dependent and developmentally regulated, and that ignoring the environmental context will miss many important associations and clues to pathogenesis. We define 'environment' broadly to include the in utero environment, maternal affection status and sex, and propose that epigenetic mechanisms are the link between our genes and our environment. PMID- 16214316 TI - Regulation of allergy by Fc receptors. AB - The aggregation of high-affinity IgE receptors (FcepsilonRI) on mast cells and basophils has long been known as the critical event that initiates allergic reactions. Monomeric IgE was recently found to induce a variety of effects when binding to FcepsilonRI. Upregulation of FcepsilonRI only requires binding, whereas other responses require FcepsilonRI aggregation. Interestingly, FcepsilonRI aggregation has recently been understood to generate a mixture of positive and negative intracellular signals. Mast cells and basophils also express low-affinity and, under specific conditions, high-affinity IgG receptors. When co-engaging these receptors with FcepsilonRI, IgG antibodies can amplify or dampen IgE-induced mast cell activation. On the basis of these findings, it has been proposed that FcRs can be used as targets and/or tools for new therapeutic approaches to allergies. PMID- 16214317 TI - Is celiac disease an autoimmune disorder? AB - Celiac disease, which results from an immune reaction to ingested cereal gluten proteins, has several autoimmune features. In particular, celiac disease patients produce highly disease specific IgA and IgG autoantibodies to tissue transglutaminase when they are on a gluten-containing diet, and they have small intestinal intraepithelial lymphocytes which can mediate direct cytotoxicity of enterocytes expressing MIC molecules in an antigen non-specific manner. Similar to typical autoimmune disorders, celiac disease has a multifactorial aetiology with complex genetics, and several autoimmune diseases are commonly presented by patients with celiac disease. Much has been learned about the immunology of celiac disease in recent years, and there is overwhelming evidence that the immune response to gluten is central to the pathogenesis. In light of this, the many autoimmune phenomena associated with celiac disease are thought-provoking, and they challenge us to rethink the boundaries between autoimmunity and immunopathology. PMID- 16214318 TI - Shaping the T cell repertoire to a bona fide autoantigen: lessons from autoimmune gastritis. AB - Murine autoimmune gastritis is one of the most well-defined organ-specific autoimmune diseases. CD4(+) T cells, which mediate the disease, recognize the highly abundant gastric H(+)/K(+) ATPase heterodimer. The H(+)/K(+) ATPase alpha subunit is also expressed in the thymus, in an aire-independent manner, whereas the H(+)/K(+) ATPase beta subunit is absent from the thymus. Analysis of both H(+)/K(+) ATPase-specific T cell receptor transgenic mice with different affinities for the gastric antigen and mice deficient in the H(+)/K(+) ATPase subunits has provided information on thymic and peripheral selection events. The H(+)/K(+) ATPase antigens play an important role in purging the repertoire of gastritogenic T cells, and recent data have suggested that this tolerance induction occurs primarily in the periphery. The gastritis system provides a powerful approach to determine the impact of peripheral antigen presentation in the target organ draining lymph node on tolerance and autoimmune disease. PMID- 16214320 TI - CD3-specific antibodies restore self-tolerance: mechanisms and clinical applications. AB - The treatment of autoimmune diseases using conventional chemical immunosuppressants has short-term effects, imposing the need for chronic treatment with its risks of over-immunosuppression. CD3-specific monoclonal antibodies can restore self-tolerance in a durable fashion after a single short term treatment, as demonstrated in several experimental models and clinically in recent-onset insulin-dependent diabetes. Disease remission involves first an immediate 'freezing' of the autoimmune response, which is linked to CD3-specific antibody-induced antigenic modulation of CD3-TCR complex at the T lymphocyte surface, followed by 'resetting' of TGF-beta-dependent T-cell mediated immunoregulation. Tolerance induction is demonstrated by persisting disease protection in spite of recovery of full immunocompetence to unrelated antigens. PMID- 16214319 TI - Animal models of arthritis caused by systemic alteration of the immune system. AB - Animal models are instrumental in understanding the etiology and pathogenetic mechanisms of rheumatoid arthritis. Several new mouse models have either been produced, including transgenics, gene-knockouts, and gene knock-ins, or established as a spontaneous disease due to natural gene mutations. These models are suitable for addressing the roles of T cells, autoantibodies, cytokines and innate immunity in the development and progression of rheumatoid arthritis. In particular, they now provide insights into how systemic alterations of the immune system result in a local development of chronic arthritis that leads to joint destruction. PMID- 16214321 TI - Apoptosis and autoimmunity. AB - Autoimmune diseases reflect the confluence of genetic, environmental and stochastic events. Recent studies have implicated apoptotic cell death pathways in initiating and propagating autoimmune diseases, as well as in rendering individuals susceptible to such diseases. Similar to autoimmunity, apoptosis is a multistep process, affecting immune and target cells, integrating numerous intrinsic and extrinsic signals, and requiring the actions of multiple gene products. Particularly relevant to the complexity of autoimmunity are the recent observations that apoptotic death might provide a primary source of tolerogen to shape the immune repertoire, or be the target of the immune response in autoimmunity, and that apoptosis is both required for lymphocyte selection and immunoregulation, and is a prominent outcome of immune and inflammatory effector pathways. PMID- 16214322 TI - Association between cytokines in induced sputum and severity of chronic obstructive pulmonary disease. AB - Cytokines are known to be increased in induced sputum in chronic obstructive pulmonary disease (COPD). In this study, the relationship between the levels of interleukin-6 (IL-6), interleukin-8 (IL-8), and tumour necrosis factor-alpha (TNF alpha) in induced sputum of patients with exacerbation of COPD, and the severity of the disease, pulmonary function tests (PFT), arterial blood gases (ABG) were studied. Twenty-four patients with exacerbation of COPD were included in the study. The patients were grouped according to their PFT into two as: Group 1 (FEV1 below 50% of the predicted value, severe-very severe COPD, n=12) and, Group 2 (FEV1 above 50% of the predicted value, mild-moderate COPD, n=12). The levels of IL-6, IL-8 and TNF-alpha in induced sputum of the subjects were measured. The mean levels of IL-6, IL-8 and TNF-alpha in induced sputum were found to be higher in Group 1 (severe-very severe COPD) than in Group 2 (mild-moderate COPD). The differences in IL-6 and IL-8 levels between groups were statistically significant (P<0.05). A significant correlation was observed between the IL-6 value and FEV(1) (r=-0.435, P=0.034), FEV1/FVC (r=-0.446, P=0.029), PaO2 (r=-0.711, P=0.000), SaO2 (r=-0.444, P=0.030) and disease duration (r=0.427, P=0.037), respectively. Also, the level of IL-8 in induced sputum was inversely correlated with FEV1 (r=-0.562, P=0.004), PaO2 (r=-0.540, P=0.006) and SaO2 (r=-0.435, P=0.034). However, all three cytokines were positively correlated with the smoking load (r=0.653, P=0.001; r=0.439, P=0.032; r=0.649, P=0.001). We conclude, therefore, that in exacerbated COPD cases with greater degrees of obstruction of the airways have higher levels of cytokines in induced sputum. This can be interpreted to mean that these cytokines are related to the clinical parameters like the ABG and PFT and seem to be the determinant of the severity of the disease. PMID- 16214323 TI - Prevalence and clinical significance of a patent foramen ovale in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: A patent foramen ovale (PFO) is not widely recognized as a factor contributing to hypoxemia in patients with chronic obstructive pulmonary disease (COPD). We therefore sought to clarify the prevalence and clinical significance of a PFO in patients with COPD, and to analyze the factors related to its occurrence. METHODS: This study included 52 consecutive stable patients with COPD and 50 healthy controls. The demographic and clinical features of the study group were noted. To test for a PFO, standard and contrast transthoracic echocardiographic examinations were performed while resting and during the Valsalva maneuver (VM). Patients performed 6-min walking tests (6 MWT), and the distances traveled were measured. RESULTS: During VM, we detected a PFO in 23 COPD patients and 10 healthy controls (P<0.01). A PFO was detected while resting in 11 COPD patients, but in none of the controls (P=0.001). Comparison of multiple parameters between COPD patients with and without a PFO during VM did not reveal any clinically significant differences. When we compared COPD patients with and without a PFO during resting, however, we found that the former had longer durations of disease, lower PaO2 and SaO2, higher dyspnea scores, shorter distances walked during 6 MWT and higher desaturation rates (P<0.05). Logistic regression analysis showed that longer duration of disease, lower SaO2 and higher systolic pulmonary artery pressure were independent predictors of the occurrence of a PFO in resting COPD patients. CONCLUSIONS: The prevalence of a PFO is higher in patients with COPD than in healthy individuals. The presence of a PFO while resting may contribute significantly to the deterioration of arterial oxygenation and performance status. These findings indicate that a PFO may be a principle cause of hypoxemia in patients with COPD. PMID- 16214324 TI - Global effects of vitamin A deficiency on gene expression in rat liver: evidence for hypoandrogenism. AB - Vitamin A (retinol) metabolites are ligands for transcription factors that regulate many genes. The liver is the main storage depot for retinol and plays a role in vitamin A homeostasis. To better understand the effects of vitamin A deficiency on liver gene expression, we produced retinol deficiency in male rats by feeding a diet low in retinol for 53 days after weaning and examined the effects on gene expression in liver using Affymetrix oligonucleotide microarrays. We detected expression of 41% of the 8799 probe sets represented on the RGU-34A GeneChips. Vitamin A deficiency resulted in major changes in liver gene expression: 805 genes (22% of all genes detected) differed at P200 copies/10(3) cells. The longitudinal analysis performed on follow-up samples showed that in cases the progression to CIN2/3 was linked to HPV16 burden increasing over time, whereas in controls a decrease of at least 1 log HPV16 DNA load was observed over>or=2 time points. CONCLUSIONS: These results show that kinetics of HPV load, rather than a single HPV detection, might be more reliable to estimate whether a HPV infection will progress or be cleared. PMID- 16214398 TI - Development and evaluation of a real-time RT-PCR assay on the LightCycler for the rapid detection of enterovirus in cerebrospinal fluid specimens. AB - BACKGROUND: Detection of enteroviral nucleic acid in cerebrospinal fluid (CSF) specimens has been demonstrated to improve the management of patients with aseptic meningitis. OBJECTIVE: To develop on the LightCycler (LC) instrument a real-time RT-PCR assay based on TaqMan technology for the detection of enteroviruses (EV) in cerebrospinal fluid (CSF) specimens. STUDY DESIGN: After evaluation of the analytical performances, seventy-four CSF samples collected prospectively from patients who have been suspected for a clinical diagnosis of meningitis were evaluated by two LC real-time RT-PCR assays and one conventional RT-PCR assay. RESULTS: Our assay detected all 30 different EV species tested, whereas no reactivity was observed with other neurotropic viruses. The analytical sensitivity of both LC RT-PCR real-time assays was 1 TCID50 for LC one-step and two-step RT-PCR assays. Results for LC one-step and LC two-step RT-PCR were compared to results of the conventional RT-PCR: of the 74 CSF specimens tested, 11 were positive and 56 were negative by all methods. Four other specimens were positive for EV by at least two of the methods (including the LC two-step RT-PCR and the conventional RT-PCR), two other CSF specimens were positive by the LC two step RT-PCR assay only, and another one CSF specimen was positive by the LC one step RT-PCR assay only. No CSF specimens were negative by the LC two-step RT-PCR assay and positive by the conventional RT-PCR assay. The sensitivity, specificity, positive and negative predictive values of both LC RT-PCR assays by using conventional RT-PCR as the "gold standard" were, respectively, 73.3, 98.3, 91.7, 93.5% for the LC one-step RT-PCR and 100, 96.6, 88.2, 100% for the LC two step RT-PCR. There was substantial agreement between the three assays (k=0.80). CONCLUSIONS: The LC two-step RT-PCR assay is a rapid, sensitive and reliable method which can be routinely performed with CSF samples for diagnosis of EV infection and is an important improvement for optimal patient management. PMID- 16214399 TI - Acyl coenzyme A dependent retinol esterification by acyl coenzyme A: diacylglycerol acyltransferase 1. AB - We provide biochemical evidence that enzymes involved in the synthesis of triacylglycerol, namely acyl coenzyme A:diacylglycerol acyltransferase (DGAT) and acyl coenzyme A:monoacylglycerol acyltransferase (MGAT), are capable of carrying out the acyl coenzyme A:retinol acyltransferase (ARAT) reaction. Among them, DGAT1 appears to have the highest specific activity. The apparent K(m) values of recombinant DGAT1/ARAT for retinol and palmitoyl coenzyme A were determined to be 25.9+/-2.1 microM and 13.9+/-0.3 microM, respectively, both of which are similar to the values previously determined for ARAT in native tissues. A novel selective DGAT1 inhibitor, XP620, inhibits recombinant DGAT1/ARAT at the retinol recognition site. In the differentiated Caco-2 cell membranes, XP620 inhibits approximately 85% of the Caco-2/ARAT activity indicating that DGAT1/ARAT may be the major source of ARAT activity in these cells. Of the two most abundant fatty acyl retinyl esters present in the intact differentiated Caco-2 cells, XP620 selectively inhibits retinyl-oleate formation without influencing the retinyl palmitate formation. Using this inhibitor, we estimate that approximately 64% of total retinyl ester formation occurs via DGAT1/ARAT. These studies suggest that DGAT1/ARAT is the major enzyme involved in retinyl ester synthesis in Caco-2 cells. PMID- 16214400 TI - Habituation of sympathetic sudomotor and vasomotor skin responses: neural and non neural components in healthy subjects. AB - OBJECTIVE: Sympathetic skin response (SSR) and skin vasomotor response (SVR) habituation was thought to be induced by neural mechanisms. Here, we investigate the hypothesis that non-neural mechanisms could also be involved. METHODS: We recorded sympathetic skin nerve activity (SSNA) from median nerve by microneurography and the corresponding SSR and SVR in 16 healthy subjects. Superficial electrical stimulation of the opposite median nerve was used to induce arousal responses. RESULTS: Throughout stimulation, SSNA, SSR and SVR amplitude showed a significant reduction. During the first ten stimuli, SSNA showed a marked decrease highly correlated to SSR and SVR changes. During the subsequent 20 stimuli SSNA did not change whereas SSR and SVR significantly decreased. SVR was significantly influenced by skin temperature changes. CONCLUSIONS: Both neural and non-neural mechanisms are involved in SSR and SVR habituation. The neural mechanisms were predominant during the first part of stimulation whereas non-neural mechanisms prevailed during the last part of stimulation. SIGNIFICANCE: During repeated arousal stimuli SSR and SVR amplitude changes did not reflect the strength of the corresponding sympathetic nerve traffic and must be interpreted with caution. PMID- 16214402 TI - Trends in European CN. PMID- 16214401 TI - Dipole localization using simulated intracerebral EEG. AB - OBJECTIVE: In the clinical interpretation of intracerebral EEGs, epileptic foci are commonly identified by visually analyzing the amplitude of the potentials. This is potentially misleading since electrodes record activity from several sources, but the nearest ones generate large amplitudes that can overpower distant sources. Our objective was to improve foci detection in intracerebral recordings by applying source localization methods. METHODS: Data were simulated by placing 3 sources in a semi-infinite medium near 3 intracerebral electrodes. Potentials were generated and contaminated with white and correlated noise. Two inverse problem algorithms, beamforming and RAP-MUSIC, were used to calculate equivalent dipoles. RESULTS: Simulations for each noise types showed that the two methods detected the source locations accurately, with RAP-MUSIC reporting lower orientation errors. With correlated noise, beamforming reconstructed original source waveforms poorly. A spatial resolution analysis was performed, in which beamforming adequately distinguished sources separated by 1.2 cm, whereas RAP MUSIC separated sources as close as 0.4-0.6 cm. CONCLUSIONS: Both source localization methods proved useful in detecting the location of dipolar sources based on simulated intracerebral potentials. For all simulations, RAP-MUSIC was more accurate than beamforming. SIGNIFICANCE: It is possible to use source localization methods traditionally applied to scalp recordings for improving source detection from intracerebral recordings. PMID- 16214403 TI - Atypical motor unit potentials in Emery-Dreifuss muscular dystrophy (EDMD). AB - OBJECTIVE: The aim of the study was to analyse electromyographic changes in Emery Dreifuss muscular dystrophy (EDMD) that are atypical for myopathy. Our special interest was focused on high amplitude polyphasic motor unit potentials (MUPs), also termed irregular MUPs. METHODS: We studied 21 EDMD patients with the diagnosis based on clinical data, DNA analysis and immunohistochemical muscle studies. Rectus femoris muscle biopsies were investigated in all affected patients. Electrophysiological investigations involved quantitative concentric needle electromyography (CNEMG) of biceps brachii (BB) and rectus femoris (RF) muscles. Simulation studies were performed to approximate the number, diameter and distribution of muscle fibers, which contribute to irregular MUPs. RESULTS: The EMG data in EDMD were compatible with myopathy. Irregular MUPs showed longer duration, larger area, size index and higher amplitude then simple ones (P < 0.05). The approximation of features of muscle fibers contributing to irregular MUP also indicated smaller (<45 microm) and larger (>55 microm) diameters than normal (50 +/- 5 microm). Muscle biopsy specimens revealed the variable muscle fiber size due to atrophy, hypertrophy, and muscle fiber splitting. CONCLUSIONS: Irregular MUPs recorded in EDMD are due to hypertrophied and atrophied fibers as well as increased fiber density. They reflect reorganization of the motor unit in a slow progression myopathic process (muscle fiber hypertrophy and splitting). SIGNIFICANCE: Irregular MUPs in EDMD most probably reflect increased variability of the muscle fiber size. PMID- 16214404 TI - The face-sensitive N170 and VPP components manifest the same brain processes: the effect of reference electrode site. AB - OBJECTIVE: (1) To investigate the hypothesis that the vertex positive potential (VPP) and the N170 ERP components reported in the face processing literature are two manifestations of the same brain processes whose relative amplitude in a given experiment is dependent on reference electrode; (2) to investigate whether differences in face/object results reported in studies looking at the VPP and N170 are attributable to the location of reference. METHODS: EEG was recorded from 53 scalp electrodes referenced online to the left mastoid while subjects viewed face, car and word stimuli. Off-line, the data were systematically re referenced to the common average, averaged mastoids, averaged earlobes, non cephalic, and nose. The correlation of timing, amplitude, and effects was investigated across reference electrodes. RESULTS: (1) The amplitude of the N170 and VPP components varies in a precisely inverse manner across reference; (2) the peaks of the N170 and VPP are temporally coincident for all reference electrodes, (3) both components can be accounted for by the same dipolar configuration, and (4) the components show identical functional properties. CONCLUSIONS: The VPP and N170 are two 'faces' of the same brain generators. SIGNIFICANCE: The differential N170/VPP effects observed in ERP studies can be accounted for by differences in reference methodology. PMID- 16214405 TI - Iliopsoas: a new electromyographic technique and normal motor unit action potential values. AB - OBJECTIVE: To describe a reliable technique of needle electrode examination and present the normal values of motor unit action potential (MUAP) parameters in iliopsoas muscle. METHODS: Thirty-one normal subjects underwent quantitative electromyographic (QEMG) examination of the iliopsoas muscle, following an ultrasonographically confirmed technique of needle electrode insertion and sampling. The leg under examination was flexed, abducted and externally rotated at the hip joint and also flexed at the knee joint. A slight flexion at the hip joint was used to uncover iliopsoas from the overlying sartorius. This provides enough space at the inguinal region between the sartorius and the femoral neurovascular bundle. Mean and outlier values of MUAP parameters and polyphasia were calculated. RESULTS: Our technique was easy to perform and secure in sampling iliopsoas. The mean +/- SD values for MUAP duration, amplitude, area, area to amplitude ratio, phases and turns were 11.5 +/- 1.35 ms, 419 +/- 71.5 microV, 633 +/- 142.7 microV ms, 1.57 +/- 0.25, 3.1 +/- 0.32, and 2.9 +/- 0.44, respectively. Lower and upper outlier limits for duration, amplitude, area and area to amplitude ratio were 3.6/20.7 ms, 150/930 microV, 100/1567 microV ms, and 0.35/3.07. Mean polyphasia was 12.6% (range 0-30%). CONCLUSIONS: The suggested EMG technique helped to distinguish iliopsoas from sartorius and at the same time increased the accessibility of its anterior surface. Normal values and outlier limits of the MUAP parameters of iliopsoas have been quantitatively established. SIGNIFICANCE: This new technique and the normal MUAP values might prove helpful for the examination of iliopsoas, important in the assessment of many neuropathic and, especially, myopathic processes. PMID- 16214406 TI - Continuous motor unit activity syndromes: a video-polysomnographic study. AB - OBJECTIVE: To ascertain the presence of abnormalities of sleep in patients with continuous motor unit activity with and without symptoms of central nervous system involvement. METHODS: Five patients with isolated neuromyotonia (Isaacs' syndrome) and 1 patient with Morvan syndrome underwent 24-h videopolysomnographic recording to investigate sleep structure, motor activities and autonomic variables during sleep. RESULTS: Macro- and microstructural organization of sleep and of the attending autonomic variables were substantially normal in patients with Isaacs' syndrome. On the contrary, sleep structure was severely disrupted with subcontinuous dream enactment and hallucinations in the patient with Morvan syndrome. The pattern of the neuromyotonic discharges, however, was not different between the patients with Isaacs' syndrome compared to Morvan syndrome, the EMG discharges persisting throughout the 24 h of recording and affecting wakefulness and sleep equally. CONCLUSIONS: Neuromyotonia is compatible with normal organization of sleep. The severe sleep abnormalities observed in Morvan syndrome cannot be simply attributed to the effects of neuromyotonia of peripheral origin. SIGNIFICANCE: Even though neuromyotonia is common to both Isaacs' and Morvan syndromes, the two conditions differ significantly in regard to CNS involvement with sleep abnormalities and lumping the two conditions together is not justified on clinical and neurophysiological grounds. PMID- 16214407 TI - Electroencephalographic background desynchronization during cerebral blood flow reduction. AB - OBJECTIVE: The aim of this study is to evaluate the usefulness of a spectral function detecting cerebral hypoperfusion. METHODS: Continuous electroencephalographic monitoring was employed during 47 consecutive carotid endarterectomies. Patients were assigned to 3 different groups according to the entity of electroencephalographic changes during carotid clamping (major changes: group A; moderate changes: group B; no change: group C). The desynchronization function, indicating the reduction of the 8-15 Hz band power, and the desynchronization index were calculated. RESULTS: Group A function decreased within 20s from clamping, with a constant slope (7.14). Desynchronization indexes were: 76.85% (group A), 40.23% (group B) and 15.29% (group C). Difference among groups was statistically significant (P < 0.0001). A case of syncope due to asystole is also reported, showing the same pattern in the descending phase of the function. CONCLUSIONS: The stereotyped time course of the desynchronization function describes the cerebral reaction to significant blood flow reduction. Values of desynchronization index exceeding 65% seem to correctly detect patients with cerebral hypoxic risk. SIGNIFICANCE: The analysis of the 8-15 Hz band desynchronization is helpful in the evaluation of cerebral hypoperfusion during carotid endarterectomy. This method could be employed in monitoring different clinical situations of ischemia. PMID- 16214408 TI - Triple stimulation technique in patients with spinocerebellar ataxia type 6. AB - OBJECTIVE: To establish further evidence that SCA6 may not be a pure cerebellar syndrome. METHODS: Seven patients with genetically confirmed SCA6 and 9 age matched normal controls were studied. Recordings of the CMAP were obtained from the right first dorsal interosseus muscle. Transcranial magnetic stimulation of the left motor cortex was applied to the contralateral scalp with a plane figure of-8 coil. Conventional transcranial magnetic stimulation (TMS), central motor conduction time (CMCT) by F-wave method and the triple stimulation technique (TST) amplitude ratio (TST test/TST control) were investigated. RESULTS: The mean resting motor threshold and mean CMCT did not show significant differences between normal controls and patients, but the mean TST amplitude ratio was significantly smaller in patients than in controls. CONCLUSIONS: An abnormal TST represents upper motor neuron loss, central axon lesions or conduction blocks, or inexcitability in response to TMS. The lack of pathological changes in the corticospinal tract of patients with SCA6 indicates that this abnormality may be caused by crossed cerebellar diaschisis, or a functional disorder in the brain resulting from CACNA1A mutations. SIGNIFICANCE: TST is a useful method for quantifying corticospinal tract dysfunction. PMID- 16214409 TI - Diltiazem treatment prevents diastolic heart failure in mice with familial hypertrophic cardiomyopathy. AB - BACKGROUND: The cardiac troponin T I79N mutation, linked to familial hypertrophic cardiomyopathy, carries a high risk of sudden cardiac death even in the absence of significant cardiac hypertrophy. The pathology underlying this mechanism has not yet been identified. AIMS: To study the underlying mechanism of this phenomenon we characterized the left ventricular (LV) performance of transgenic mice carrying the human troponin T mutation I79N under basal and isoproterenol induced stress conditions. METHODS AND RESULTS: LV function was analyzed by recording pressure-volume loops using a microconductance catheter. Despite a hypercontractile systolic function under basal conditions TnT-I79N mice showed a diastolic dysfunction indicated by an increase in end-diastolic pressure-volume relationship (EDPVR), a load-independent factor of LV stiffness (0.06+/-0.01 vs. 0.02+/-0.01; P<0.05), when compared to mice expressing human wild-type troponin T (TnT-WT). TnT-I79N mutants developed severe diastolic heart failure and cardiac sudden death under isoproterenol stress. This was prevented after pretreatment with the L-type Ca2+ channel inhibitor diltiazem. CONCLUSIONS: Diastolic dysfunction due to increased LV stiffness in TnT-I79N mice leads to severe primary diastolic heart failure and finally to cardiac sudden death, which can be prevented by diltiazem. PMID- 16214410 TI - Gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is increasingly common worldwide; symptoms differ between individuals and endoscopically visible injury is present in only about 50% of cases. Although GERD is a disorder of gastrointestinal motility and structure, the most effective therapy is based on the use of acid antisecretory drugs. Proton pump inhibitors (PPIs), the most effective class of acid suppression agents to date, have revolutionised the management of GERD. However, PPIs do have some shortcomings and recent developments include documentation of increased healing rates with more prolonged acid suppression, more prolonged acid suppression with a new PPI (tenatoprazole) and more rapid onset of acid suppression with a new class of drugs, the reversible, potassium-competitive acid blockers. Studies with motility agents, such as the 5-HT(4) partial agonist tegaserod and the GABA(B) agonist baclofen, indicate that motility is important in the pathogenesis of GERD but, for several reasons, it will be a challenge to develop new classes of drug that outperform current PPIs with respect to efficacy, broad applicability and safety. PMID- 16214413 TI - Probiotics and nutraceuticals: non-medicinal treatments of gastrointestinal diseases. AB - The demonstration that immune and epithelial cells can discriminate between different microbial and bioactive plant species has extended the known mechanism(s) of action of nutraceuticals and probiotics beyond simple nutrition and/or antimicrobial effects. The progressive unravelling of these plant and bacterial effects on systemic immune and intestinal epithelial cell function has led to new credence for the use of probiotics and nutraceuticals in clinical medicine. Level I evidence now exists for the therapeutic use of probiotics in infectious diarrhea in children, recurrent Clostridium difficile-induced infections and post-operative pouchitis. Additional evidence is being acquired for the use of probiotics in other gastrointestinal infections, irritable bowel syndrome and inflammatory bowel disease. Not all individual probiotic strains have the same efficacy, and future clinical trials may focus on multistrain preparations agents with known efficacy. The use of nutraceuticals and probiotics as therapeutic agents for gastrointestinal disorders is rapidly moving into clinical usage. Scientific studies are providing mechanisms of action to explain the therapeutic effects, and randomized controlled trials are providing the necessary evidence for their incorporation into the therapeutic armamentarium. PMID- 16214414 TI - Hyaluronan cross-linking: a protective mechanism in inflammation? AB - Production of the glycosaminoglycan hyaluronan is increased at sites of inflammation, often correlating with the accumulation of leukocytes. Mounting evidence suggests that this polysaccharide can be organized into a wide variety of molecular architectures by its association with specific binding proteins, leading to the formation of fibrils and cable-like structures involving a large number of hyaluronan chains. We propose that hyaluronan cross-linking is part of a protective mechanism, promoting adhesion of leukocytes to the hyaluronan complexes rather than enabling contact with inflammation-promoting receptors on the underlying tissues. Leukocytes are thus maintained in a non-activated state by appropriate receptor clustering or receptor co-engagement. Additionally, hyaluronan networks might serve as scaffolds to prevent the loss of extracellular matrix components during inflammation and to sequester proinflammatory mediators. PMID- 16214415 TI - The role of stress-response systems for the pathogenesis and progression of MS. AB - Disease progression in multiple sclerosis (MS)--an inflammatory demyelinating and neurodegenerative disease with a presumed T-cell driven autoimmune origin--has long been hypothesized to be associated with stress. However, this notion has only recently been supported by prospective clinical studies. Several clinical and molecular studies in MS and its animal models have recently shown disruptions in the communication between the immune system and the two major stress response systems, the hypothalamo-pituitary-adrenal (HPA) axis and the autonomic nervous system. Insensitivity to glucocorticoid and beta-adrenergic modulation might be involved in overshooting inflammation in MS, whereas hyperactivity of the HPA axis has been linked to neurodegeneration and increased disability. Here, we integrate findings from molecular, cellular, experimental, clinical and epidemiological research to describe the involvement of stress response systems in MS pathogenesis and progression. PMID- 16214416 TI - SNOR and wheeze: the asthma enzyme? AB - Conventionally, asthma is defined as involving both airway inflammation and airway smooth muscle hyper-responsiveness. However, Que and coworkers have recently uncoupled these concepts, showing that mice lacking an S-nitrosothiol reductase have allergen-induced airway inflammation but do not have airway hyper responsiveness. These data are consistent with recent clinical evidence that: (i) S-nitrosothiol signaling is abnormal in human asthma, (ii) nitric oxide in exhaled air might be only a biomarker for the metabolism of more physiologically relevant nitrogen oxides and (iii) the biochemical response to airway inflammation is central to asthma pathophysiology. PMID- 16214417 TI - Notch and Wnt inhibitors as potential new drugs for intestinal neoplastic disease. AB - Colorectal cancer is a major cause of death in the western world. Recent advances in treatment comprise variations on the classical themes of surgical resection combined with chemotherapy using cytotoxic drugs and radiation therapy. Because this therapy is only moderately successful, novel approaches to the treatment of colorectal cancer are required. Our rapidly increasing knowledge of molecular signalling pathways that are deregulated in colorectal cancer might provide a platform from which to develop new rational cancer therapies. Here, we give an update on the roles of the Wnt and Notch signalling pathways in the self renewal of the intestinal epithelium and the consequences of Wnt deregulation in colorectal cancer. We focus on the potential of recently identified small molecule inhibitors of the Wnt pathway and gamma-secretase inhibitors of the Notch pathway as novel colon cancer therapeutics. PMID- 16214419 TI - Nucleic acid medicines move towards the clinic. PMID- 16214418 TI - DNA repair inhibition: a selective tumour targeting strategy. AB - Advanced cancer is a leading cause of death in the developed world. Chemotherapy and radiation are the two main treatment modalities currently available. The cytotoxicity of many of these agents is directly related to their propensity to induce DNA damage. However, the ability of cancer cells to recognize this damage and initiate DNA repair is an important mechanism for therapeutic resistance and has a negative impact upon therapeutic efficacy. Pharmacological inhibition of DNA repair, therefore, has the potential to enhance the cytotoxicity of a diverse range of anticancer agents. Moreover, the use of inhibitors of DNA repair or DNA damage signalling pathways appears to provide an exciting opportunity to target the genetic differences that exist between normal and tumour tissue. PMID- 16214420 TI - Identification of novel biomarkers for Niemann-Pick disease using gene expression analysis of acid sphingomyelinase knockout mice. AB - Although several therapies are available or being developed for lysosomal storage disorders (LSDs), assessment of therapeutic efficacy is challenged by the lack of markers to assess disease progression and severity. This is particularly true for rare diseases such as LSDs, since natural history data from human populations are often lacking. Herein we describe the use of gene expression analysis in the acid sphingomyelinase-deficient mouse model (ASMKO) of Types A and B Niemann-Pick disease (NPD) to identify novel serum biomarkers. We used microarray and real time PCR analyses to compare mRNA expression in ASMKO and normal mice in two important sites of pathology, lung and brain, and from these data identified and validated several potential biomarkers. The cytokine MIP-1alpha was markedly elevated in ASMKO mouse serum, and following enzyme replacement therapy (ERT) it was reduced to normal levels. Total iron levels were similarly elevated in ASMKO mice, reflective of the elevated ferritin light chain transcript, and decreased to normal after ERT. Serum growth hormone levels were also elevated in ASMKO mice and were reduced to normal after brain-directed gene therapy, but not ERT. These studies illustrate the value of gene expression analysis for the identification of biomarkers, and provide new insight into the pathobiology of NPD. PMID- 16214421 TI - Knowledge guided analysis of microarray data. AB - To microarray expression data analysis, it is well accepted that biological knowledge-guided clustering techniques show more advantages than pure mathematical techniques. In this paper, Gene Ontology is introduced to guide the clustering process, and thus a new algorithm capturing both expression pattern similarities and biological function similarities is developed. Our algorithm was validated on two well-known public data sets and the results were compared with some previous works. It is shown that our method has advantages in both the quality of clusters and the precision of biological annotations. Furthermore, the clustering results can be adjusted according to different stringency requirements. It is expected that our algorithm can be extended to other biological knowledge, for example, metabolic networks. PMID- 16214422 TI - Influence of oxygen, temperature and carbon dioxide on nitric oxide formation from nitrite as measured in expired gas from in situ perfused rabbit lungs. AB - In biological systems, nitric oxide (NO) may be generated non-enzymatically from nitrite (nitrite-derived NO), in addition to nitric oxide synthase-catalyzed (NOS derived) L-arginine-dependent formation. Through recordings of expired NO, we studied the influence of temperature on NOS- and nitrite-derived NO in the perfused lung. We also studied the impact of other influencing factors (O(2), CO(2), and pH) on nitrite-derived NO in the same system. Both NO-generating systems exhibited biphasic temperature dependence with a positive correlation between temperature and NO generation that peaked between 42 and 44 degrees C. The nitrite-derived NO generation was enhanced by hypoxia alone (>20 x after 5 min) and further by concomitant increase in CO(2). The CO(2) effect could not be explained by changes in extracellular pH and was unaltered by acetazolamide. We conclude that the temperature dependence in the known enzyme-catalyzed NOS derived NO and especially in the nitrite-derived NO strengthens the hypothesis that an enzyme could be involved in nitrite-derived NO formation. The enhancement of nitrite-derived NO by increases in CO(2) suggests that this system could be of importance to improve perfusion in ischemic tissues. PMID- 16214423 TI - Hansenula polymorpha NMR2 and NMR4, two new loci involved in nitrogen metabolite repression. AB - In the yeast Hansenula polymorpha (Pichia angusta) nitrate assimilation is tightly regulated and subject to a dual control: nitrogen metabolite repression (NMR), triggered by reduced nitrogen compounds, and induction, elicited by nitrate itself. In a previous paper [Serrani, F., Rossi, B. and Berardi, E (2001) Nitrogen metabolite repression in Hansenula polymorpha: the nmrl-l mutation. Curr. Genet. 40, 243-250], we identified five loci (NMR1-NMR5) involved in NMR, and characterised one of them (NMR1), which likely identifies a regulatory factor. Here, we describe two more mutants, namely nmr2-1 and nmr4-1. The first one possibly identifies a regulatory factor involved in nitrogen metabolite repression by various nitrogen sources alternative to ammonium. The second one, apparently involved in ammonium assimilation, probably has sensor functions. PMID- 16214424 TI - Biochemical characterization of the RECQ4 protein, mutated in Rothmund-Thomson syndrome. AB - Rothmund-Thomson syndrome (RTS) is an autosomal recessive disorder characterized by growth deficiency, skin and skeletal abnormalities, and a predisposition to cancer. Mutations in the RECQ4 gene, one of five human homologs of the E. coli recQ gene, have been identified in a subset of RTS patients. Cells derived from RTS patients show high levels of chromosomal instability, implicating this protein in the maintenance of genomic integrity. However, RECQ4 is the least characterized of the RecQ helicase family with regard to its molecular and catalytic properties. We have expressed the human RECQ4 protein in E. coli and purified it to near homogeneity. We show that RECQ4 has an ATPase function that is activated by DNA, with ssDNA being much more effective than dsDNA in this regard. We have determined that a DNA length of 60 nucleotides is required to maximally activate ATP hydrolysis by RECQ4, while the minimal site size for ssDNA binding by RECQ4 is between 20 and 40 nucleotides. Interestingly, RECQ4 possesses a single-strand DNA annealing activity that is inhibited by the single-strand DNA binding protein RPA. Unlike the previously characterized members of the RecQ family, RECQ4 lacks a detectable DNA helicase activity. PMID- 16214425 TI - Contribution of Msh2 and Msh6 subunits to the asymmetric ATPase and DNA mismatch binding activities of Saccharomyces cerevisiae Msh2-Msh6 mismatch repair protein. AB - Previous analyses of both Thermus aquaticus MutS homodimer and Saccharomyces cerevisiae Msh2-Msh6 heterodimer have revealed that the subunits in these protein complexes bind and hydrolyze ATP asymmetrically, emulating their asymmetric DNA binding properties. In the MutS homodimer, one subunit (S1) binds ATP with high affinity and hydrolyzes it rapidly, while the other subunit (S2) binds ATP with lower affinity and hydrolyzes it at an apparently slower rate. Interaction of MutS with mismatched DNA results in suppression of ATP hydrolysis at S1-but which of these subunits, S1 or S2, makes specific contact with the mismatch (e.g., base stacking by a conserved phenylalanine residue) remains unknown. In order to answer this question and to clarify the links between the DNA binding and ATPase activities of each subunit in the dimer, we made mutations in the ATPase sites of Msh2 and Msh6 and assessed their impact on the activity of the Msh2-Msh6 heterodimer (in Msh2-Msh6, only Msh6 makes base specific contact with the mismatch). The key findings are: (a) Msh6 hydrolyzes ATP rapidly, and thus resembles the S1 subunit of the MutS homodimer, (b) Msh2 hydrolyzes ATP at a slower rate, and thus resembles the S2 subunit of MutS, (c) though itself an apparently weak ATPase, Msh2 has a strong influence on the ATPase activity of Msh6, (d) Msh6 binding to mismatched DNA results in suppression of rapid ATP hydrolysis, revealing a "cis" linkage between its mismatch recognition and ATPase activities, (e) the resultant Msh2-Msh6 complex, with both subunits in the ATP bound state, exhibits altered interactions with the mismatch. PMID- 16214426 TI - Protein kinase C modulates the pulmonary inflammatory response in acute pancreatitis. AB - The present study aims at evaluating the role of protein kinase C (PKC) in the development of acute lung injury, production of inflammatory mediators and expression of adhesion molecules on leukocytes after induction of acute pancreatitis (AP). AP was induced by the intraductal infusion of 5% sodium taurodeoxycholate in the rat. The animals had the PKC inhibitor polymyxin B administered intraperitoneally 30min prior to induction of AP. Levels of protein content, protease activity, cytokines and chemokines in bronchoalveolar lavage fluid (BALF) were assessed 1 and 6h after AP induction. Adhesion molecule expression on leukocytes were measured by flowcytometry. Pretreatment with polymyxin B prevented against acute pancreatitis-induced lung injury and the otherwise occurring increases in TNF-alpha, IL-1beta, MCP-1 and IL-10, as well as against the decreases in IL-2, IFNgamma and TIMP-1, decreased protease activity and down-regulation of CD31, CD54 and CD62L on recruited neutrophils and macrophages in BALF. The results indicate that the leukocyte response in acute pancreatitis vary depending on leukocyte subpopulation. It seems that activation of the PKC signalling pathway may play an important role in pancreatitis associated lung injury. PMID- 16214427 TI - Comparison of HPLC method and commercial ELISA assay for asymmetric dimethylarginine (ADMA) determination in human serum. AB - The performance of a new ELISA assay kit (DLD Diagnostika GmbH, Hamburg, Germany) for the determination of asymmetric dimethylarginine (ADMA) was evaluated against a reversed phase HPLC method. ADMA concentrations of 55 serum samples were measured with both methods. The intra-assay CV for ADMA-ELISA was 19% (n=10). Inter-assay CVs for ADMA-ELISA were 9% for kit control 1 (0.410+/-0.037 microM) and 14% for kit control 2 (1.174+/-0.165 microM). The intra- and inter-assay CVs for HPLC assay for ADMA were 2.5% (0.586+/-0.015 microM) and 4.2% (0.664+/-0.028 microM), respectively. There was no correlation between these two methods (R(2)=0.0972). The effect of storage conditions of the samples on ADMA concentrations was investigated by HPLC. ADMA concentration was stable after four freezing and thawing cycles. Overall, the HPLC method offered better sensitivity, selectivity and, very importantly, simultaneous determination of ADMA, SDMA, l homoarginine and l-arginine. PMID- 16214428 TI - Healthcare infrastructure, contraceptive use and infant mortality in Uttar Pradesh, India. AB - This paper analyzes data on approximately 30,000 women from a survey in Uttar Pradesh in 1995 together with the data from surveys of public and private providers of healthcare and family planning services. A framework was developed for analyzing the effects of quality of services on utilization, and for understanding the gradual evolution of the healthcare infrastructure. The empirical results from logistic regressions for use of female sterilization and IUD showed significant effects of quality of services in government and private hospitals, and of socioeconomic variables such as education, caste, and an index of household possessions. Secondly, models for infant mortality of children born in the preceding 3-year period showed significant effects of socioeconomic variables, quality of healthcare services and birth spacing. Lastly, analysis of data at a more aggregated (Primary Sampling Unit) level indicated differential effects of economic development on the quality of services available in the public and private facilities. PMID- 16214429 TI - Accounting for global protein deformability during protein-protein and protein ligand docking. AB - Computational docking methods are valuable tools aimed to simplify the costly process of drug development and improvement. Most current approaches assume a rigid receptor structure to allow virtual screening of large numbers of possible ligands and putative binding sites on a receptor molecule. However, inclusion of receptor flexibility can be of critical importance since binding of a ligand can lead to changes in the receptor protein conformation that are sterically necessary to accommodate a ligand. Recent approaches to efficiently account for receptor flexibility during docking simulations are reviewed. In particular, accounting efficiently for global conformational changes of the protein backbone during docking is a still challenging unsolved problem. An approximate method has recently been suggested that is based on relaxing the receptor conformation during docking in pre-calculated soft collective degrees of freedom (M. Zacharias, Rapid protein-ligand docking using soft modes from molecular dynamics simulations to account for protein deformability: binding of FK506 to FKBP, Proteins: Struct., Funct., Genet. 54 (2004) 759-767). Test applications on protein-protein docking and on docking the inhibitor staurosporine to the apo form of cAMP-dependent protein kinase A catalytic domain indicate significant improvement of docking results compared to rigid docking at a very modest computational demand. Accounting for receptor conformational changes in pre calculated global degrees of freedom might offer a promising route to improve systematic docking screening simulations. PMID- 16214430 TI - Dynamics of signaling by PKA. AB - The catalytic and regulatory subunits of cAMP-dependent protein kinase (PKA) are highly dynamic signaling proteins. In its dissociated state the catalytic subunit opens and closes as it moves through its catalytic cycle. In this subunit, the core that is shared by all members of the protein kinase family is flanked by N- and C-terminal segments. Each are anchored firmly to the core by well-defined motifs and serve to stabilize the core. Protein kinases are not only catalysts, they are also scaffolds. One of their major functions is to bind to other proteins. In addition to its interactions with the N- and C- termini, the catalytic subunit interacts with its inhibitor proteins, PKI and the regulatory subunits. Both bind with subnanomolar affinity. To achieve this tight binding requires docking of a substrate mimetic to the active site cleft as well as a peripheral docking site. The peripheral site used by PKI is distinct from that used by RIalpha as revealed by a recent structure of a C:RIalpha complex. Upon binding to the catalytic subunit, the linker region of RIalpha becomes ordered. In addition, cAMP-binding domain A undergoes major conformational changes. RIalpha is a highly malleable protein. Using small angle X-ray scattering, the overall shape of the regulatory subunits and corresponding holoenzymes have been elucidated. These studies reveal striking and surprising isoform differences. PMID- 16214432 TI - Characterization of a mouse amelogenin [A-4]/M59 cell surface receptor. AB - Amelogenin proteins comprise up to 90% of the organic matrix of developing enamel in the vertebrate tooth. Alternative splicing of mouse amelogenin pre-mRNA leads to the production of more than 14 protein isoforms, the functions of which are not totally understood. The smaller splice products, [A + 4] or M73 and [A - 4] or M59, have been shown to act differently as signaling molecules affecting odontogenic and other cell types. The mechanisms of these signaling processes, beginning with receptor identification, are not well understood. Utilizing radiolabeled [A - 4], we show here that 3H[A - 4] binds in a saturable fashion to the cell surface of C2C12 mouse fetal myoblasts at 4 degrees C, and not only binds at the surface but is internalized at 37 degrees C. "Far Western" immunohistochemistry performed on sections of E18 mouse incisors and molars with biotin-labeled [A - 4] as the primary ligand demonstrates [A - 4]-biotin binding to polarizing ameloblasts and odontoblasts, cells of the dental follicle, and along the stratum intermedium. Using [A - 4] affinity column chromatography and [A - 4]-biotin label transfer reaction, we have identified a 95 kDa C2C12 cell surface protein which bound [A - 4]. Utilizing Tandem MS (MS/MS) sequencing, we report the novel finding of the 95 kDa murine transmembrane protein, LAMP-1, originally identified as a lysosomal membrane protein that is also found at the cell surface, as an [A - 4] cell binding protein. PMID- 16214431 TI - How does human stem cell therapy influence gene expression after liver injury? Microarray evaluation on a rat model. AB - BACKGROUND: Tissue homeostasis is guaranteed by stem proliferating reserve, depending on dynamic changes in gene expression. A high plasticity is shown by the haematopoietic stem cells, potential source for liver regeneration. AIM: We aimed to evaluate the gene expression modifications induced by human haematopoietic stem cell therapy after liver injury in rats. SUBJECTS: Rats were sorted as follows: (A) human-haematopoietic stem cell injection after allyl alcohol liver damage; (B) only haematopoietic stem cell injection; (C) only allyl alcohol injection; and (D) sacrifice without any treatment. METHODS: Livers, spleens and bone marrows were analysed with flow-cytometry. Livers were also studied by reverse-transcription PCR, histology, immunohistochemistry and microarray analysis; selected genes were confirmed by real-time PCR. RESULTS: In subset A, haematopoietic stem cells were selectively recruited by liver, with respect to the group B, and they improved the liver regeneration process compared to group C. As regards microarrays, haematopoietic stem cell infusion upregulates 265 genes and downregulates 149 genes. Differentially regulated genes belong to a broad range of functional pathways, including proliferation, differentiation, adhesion/migration and transcripts related to oval-cell activation. Real-time PCR validated array results. CONCLUSIONS: Our study confirmed the capacity of haematopoietic stem cells to contribute to liver regeneration. Moreover, microarray analysis led to the identification of genes whose regulation strongly correlates with a more efficient process of liver repair after haematopoietic stem cell injection. PMID- 16214433 TI - High bone turnover of type I collagen depends on fetal growth. AB - The bone metabolic processes of proliferation and differentiation in preterm and term newborns have yet to be fully elucidated. Seventy-four umbilical cord blood samples were collected from preterm and term newborns delivered at 27 to 42 gestational weeks (GWs). Carboxy-terminal propeptide of type I procollagen (PICP), pyridinoline cross-linked telopeptide domain of type I collagen (ICTP), alkaline phosphatase (ALP), and bone-specific alkaline phosphatase (BAP) were measured. Calcitonin (CT), estrogen (E2), intact parathyroid hormone, and insulin like growth factor-I (IGF-I) were also examined in 20 or 23 randomly selected samples. We conducted cross-sectional regression analyses for bone metabolic markers, fetal growth markers including GWs, birth weight (BW), height (BH) and head circumference (HC), and bone related hormones. PICP and ICTP activities were very high, but decreased significantly with fetal growth based on GWs, BW, BH, and HC changes (GWs, BW, and BH to both PICP and ICTP, P < 0.0001; HC to ICTP, P < 0.0001; HC to PICP, P < 0.05), while BAP and ALP did not change significantly. E2 and CT both showed a significant positive correlation with Ca (P < 0.05), but neither hormone had any apparent correlation with PICP, ALP, BAP, or ICTP. These results suggest very active bone formation and resorption of type I collagen to be dependent on fetal growth and that fetal osteoblasts dominate the proliferation phase of development rather than the maturation phase. However, factors contributing to high bone turnover in the fetus remain to be elucidated. PMID- 16214434 TI - Usefulness of ST-segment elevation in the inferior leads in predicting ventricular septal rupture in patients with anterior wall acute myocardial infarction. AB - The ventricular septum receives its blood supply from the septal perforators of the left anterior descending (LAD) coronary artery and the right coronary artery. However, when the LAD artery extends to the inferior wall, beyond the apex (so called wrapped LAD), the ventricular septum near the apex receives blood supply only from the LAD artery. As a consequence, ventricular septal rupture (VSR) would seem more likely in myocardial infarction with occlusion of this type of LAD artery. To test this hypothesis, we compared electrocardiographic findings in 21 patients who had anterior acute myocardial infarction that was complicated by VSR with those in 275 patients who had acute myocardial infarction that was not complicated by VSR. We observed ST-segment elevation in all inferior leads (II, III, and aVF) in addition to anterior leads in 42.9% of patients (9 of 21) who had VSR but in only 3.6% of those (10 of 275) who did not have VSR. Abnormal Q waves appeared in all 3 inferior leads in 44.4% of patients (8 of 18) who had VSR but in only 4.0% of those (10 of 250) who did not have VSR. Thus, the incidence of ST-segment elevation and abnormal Q waves in the inferior leads was significantly (p <0.001) greater in the VSR group. In addition, multivariate analysis of patient characteristics, including advanced age, female gender, and coronary morphology, showed VSR to be significantly correlated with ST-segment elevation (odds ratio 16.93, 95% confidence interval 4.13 to 69.30) and abnormal Q waves (odds ratio 13.64, 95% confidence interval 3.16 to 58.79) in the 3 inferior leads. In conclusion, these electrocardiographic findings can be useful predictors of complication by VSR. PMID- 16214435 TI - Racial and ethnic differences in the treatment and outcome of cardiogenic shock following acute myocardial infarction. AB - We investigated the association between race/ethnicity on the use of cardiac resources in patients who have acute myocardial infarction that is complicated by cardiogenic shock. The Should We Emergently Revascularize Occluded Coronaries for Cardiogenic Shock (SHOCK) trial examined the effect of reperfusion and revascularization treatment strategies on mortality. Patients screened but not enrolled in the SHOCK Trial (n = 1,189) were entered into the SHOCK registry. Of the patients in the United States registry (n = 538) who had shock due to predominant left ventricular failure, 440 were characterized as white (82%), 42 as Hispanic (8%), 34 as African-American (6%), and 22 as Asian/other (4%). The use of invasive procedures differed significantly by race/ethnicity. Hispanic patients underwent coronary angiography significantly less often than did white patients (38 vs 66%, p = 0.002). Among those patients who underwent coronary angiography, there were no race/ethnicity differences in the proportion of patients who underwent revascularization (p = 0.353). Overall in-hospital mortality (57%) differed significantly by race/ethnicity (p = 0.05), with the highest mortality rate in Hispanic patients (74% vs 65% for African-Americans, 56% for whites, and 41% for Asian/other). After adjustment for patient characteristics and use of revascularization, there were no mortality differences by race/ethnicity (p = 0.262), with all race/ethnicity subgroups benefiting equally by revascularization. In conclusion, the SHOCK registry showed significant differences in the treatment and in-hospital mortality of Hispanic patients who had cardiogenic shock, with these patients being less likely to undergo percutaneous coronary intervention. Therefore, early revascularization should be strongly considered for all patients, independent of race/ethnicity, who develop cardiogenic shock after acute myocardial infarction. PMID- 16214436 TI - Short- and long-term outcomes of patients with electrocardiographic left ventricular hypertrophy after fibrinolysis for acute myocardial infarction. AB - There is conflicting evidence with regard to the value of electrocardiographic left ventricular hypertrophy (LVH) in myocardial infarction. Of 5,951 patients in the ASSENT-3 trial, 273 (5%) had LVH on baseline electrocardiograms and had significantly higher mortality rates at 30 days (11% vs 6%, p = 0.001) and 1 year (13% vs 8%, p = 0.007). After adjustment for differences in baseline parameters, LVH remained an independent predictor of 30-day (hazard ratio 2.3, 95% confidence interval 1.4 to 3.9) and 1-year (hazard ratio 1.8, 95% confidence interval 1.1 to 2.8) mortality rates. Thus, electrocardiographic LVH is a prognostic tool in identifying short- and long-term mortality rates in patients who have ST elevation myocardial infarction and receive fibrinolysis. PMID- 16214438 TI - Usefulness of bone mineral density to predict significant coronary artery disease. AB - Low bone mineral density (BMD) and coronary artery disease (CAD) share common risk factors. To investigate whether low BMD (osteoporosis and/or osteopenia) independently predicts CAD compared with traditional cardiovascular risk factors, a retrospective analysis was performed in consecutive ambulatory patients (n = 209, 89% women) who underwent dual-energy x-ray absorptiometry and coronary angiography within the same 12-month period. Angiograms were classified as showing significant CAD if > or =50% luminal narrowing in a major coronary artery was noted. Clinical variables associated with CAD (age, hypertension, diabetes, high fasting glucose level, smoking, family history of CAD, and dyslipidemia) were examined. Dual-energy x-ray absorptiometric scans were classified based on World Health Organization criteria: normal (T score >-1.0 SD), osteopenia (T score -1.0 to -2.5 SD), and osteoporosis (T score <-2.5 SD). Univariate and multivariate analyses were employed to determine whether low BMD independently predicts CAD. Univariate predictors of CAD were hypertension, smoking, diabetes, high fasting glucose level, dyslipidemia, family history of CAD, and low BMD. Multivariate predictors were hypertension, family history of CAD, fasting glucose level, and osteoporosis. Odds ratio for the prediction of angiographically documented CAD was highest for osteoporosis (odds ratio 5.6, 95% confidence interval 2.6 to 12.0, p <0.0001). In conclusion, low BMD appears to independently predict significant CAD in women, with a higher odds ratio than traditional risk factors. Our study is the first to report osteoporosis as a predictor of angiographically proved CAD in a population predominantly of women. PMID- 16214437 TI - Effects of glucose-insulin-potassium infusion on ST-elevation myocardial infarction in patients treated with thrombolytic therapy. AB - The role of glucose-insulin-potassium (GIK) infusion in the management of acute myocardial infarction is not well established. This prospective, randomized study comprised 120 patients who had ST-elevation myocardial infarction that was treated within 12 hours from symptom onset with a high dose of GIK (25% glucose, 50 IU of soluble insulin per liter, and 80 mmol of potassium chloride per liter at 1 ml/kg/hour over 24 hours) as adjunct to thrombolytic therapy (1.5 MU of streptokinase/30 to 60 minutes; GIK group) or thrombolytic therapy alone (control group). The primary end point of the study was the rate of major adverse cardiac events (MACEs) at 1 month, defined as a composite of cardiac death, reinfarction, serious arrhythmias (ventricular fibrillation and/or tachycardia), and severe heart failure. The secondary end points were the rate of MACEs at 1 year and improvement in left ventricular systolic function. The incidence of MACEs at 1 month was significantly lower in the GIK group (10% vs 32.5%, relative risk 0.24, 95% confidence interval 0.09 to 0.63, p = 0.0043). Patients in the GIK group had significant decreases in ventricular tachycardia and/or fibrillation (1.3% vs 15.0%, p = 0.003) and severe heart failure (3% vs 12.5%, p = 0.031). The rate of MACEs at 1 year was also significantly lower in the GIK group (13% vs 40.0%, relative risk 0.22, 95% confidence interval 0.09 to 0.55, p = 0.0012). After 1 year, there was a significant improvement in left ventricular ejection fraction in the GIK group (from 48 +/- 8% to 51 +/- 10%, p <0.01), which was not observed in the control group. In conclusion, high-dose GIK, used as an adjunct to thrombolytic therapy, was safe and improved clinical outcome at 1 month. The beneficial effect of GIK infusion was maintained up to 1 year. PMID- 16214439 TI - Myocardial perfusion study of panic attacks in patients with coronary artery disease. AB - Panic disorder (PD) and panic-like anxiety have been associated with an increased risk of cardiovascular death. No study has specifically examined the association between panic attacks and ischemia in patients who have coronary artery disease (CAD). We hypothesized that panic attacks would induce myocardial perfusion defects in patients who have CAD and PD. Sixty-five patients who had CAD and positive results with nuclear exercise stress testing (35 with PD and 30 without PD served as controls) underwent a well-established panic challenge test (1 vital capacity inhalation of a gas mixture containing 35% carbon dioxide and 65% oxygen) and were injected with technetium-99m sestamibi at inhalation. Single photon emission computed tomography was used to assess per-panic challenge perfusion defects, and heart rate, blood pressure, and 12-lead electrocardiogram were continuously measured during the procedure. Patients were not withdrawn from their cardiac medications. Patients who had PD were significantly younger than the controls; otherwise groups did not differ with respect to gender, cardiac medications, nuclear exercise test results, and baseline heart rate and blood pressure. Seventy-four percent of patients (26 of 35) who had PD had a panic attack at inhalation versus 6.7% of controls (2 of 30, p <0.001). As hypothesized, patients who had PD and demonstrated a panic attack were more likely to develop a reversible myocardial perfusion defect than were controls who did not have an attack (80.9% vs 46.4% p = 0.009). Thus, despite being on their cardiac medications, panic attacks preferentially induced significant perfusion defects in patients who had CAD and PD. In conclusion, panic attacks in patients who have CAD appear to be bad for the heart. PMID- 16214440 TI - Echocardiographic characteristics of electrocardiographically unrecognized myocardial infarctions in a community population. AB - Unrecognized myocardial infarction (UMI) as diagnosed by surveillance electrocardiography has been shown to carry the same poor prognosis as recognized myocardial infarction (RMI). The echocardiographic characteristics of UMI have never been studied before. Due to a similar prognosis, we hypothesized that UMI and RMI would exhibit similar degrees of echocardiographic ventricular dysfunction. We studied a random community cohort of 2,042 adults who were > or =45 years of age in a cross-sectional setting in Olmsted County, Minnesota. RMI was diagnosed by review of medical records and UMI was diagnosed if the electrocardiogram met MI criteria without a previous MI recorded in the medical record. All subjects underwent transthoracic echocardiography. We identified 80 patients who had UMI and 101 who had RMI. In bivariate analyses, a stepwise increase in echocardiographic abnormalities was observed from participants who had no MI to UMI to RMI: respective mean ejection fractions were 63%, 61%, and 55; prevalences in left ventricular enlargement were 13%, 22%, and 52%; mean left ventricular mass indexes were 98, 103, and 118 g/m(2); prevalences in regional wall motion abnormality were 2%, 13%, and 42%; and prevalences in diastolic dysfunction were 25%, 56%, and 65% (p for trend <0.0001 for all comparisons). After adjusting for standard coronary risk factors, patients who had UMI continued to exhibit significant abnormalities in systolic dysfunction, diastolic dysfunction, and regional wall motion abnormality, although to a lesser extent than patients who had RMI. In conclusion, patients who have UMI manifest structural abnormalities more commonly than do patients who have no MI but less commonly than do those who have RMI. The similar prognosis after UMI and RMI cannot be explained by comparable degrees of ventricular damage. PMID- 16214442 TI - Relation of QRS duration on the surface 12-lead electrocardiogram with mortality in patients with known or suspected coronary artery disease. AB - The aim of this study was to assess the relation between QRS duration and mortality in patients with known or suspected coronary artery disease, after adjustment for myocardial functional abnormalities, as assessed by exercise echocardiography. We studied 4,033 patients (age 62 +/- 12 years; 2,360 men, 18% with previous myocardial infarction) who underwent symptom-limited exercise echocardiography. The QRS duration was electronically measured from the 12-lead electrocardiogram. The incremental value of the QRS duration for predicting mortality was assessed by adding the QRS duration at the end of each of these modeling steps: clinical data, exercise electrocardiographic, and exercise echocardiographic variables. The QRS duration correlated positively with age, the wall motion score index at rest, and percentage of ischemic segments and negatively with workload (p = 0.0001). Of the 4,033 patients, 252 died during a median follow-up of 3 years. The QRS duration was univariately associated with an increased risk of death (relative risk 8.5, 95% confidence interval CI 4.4 to 16.4, p <0.0001). In an incremental multivariate model, the clinical predictors of mortality were age, male gender, previous infarction, and diabetes mellitus (chi-square 122). Workload was incremental to clinical data in the exercise test model (chi-square 193, p <0.0001). The exercise wall motion score index was incremental to both models (chi-square 211, p <0.001). The QRS duration was associated with an incremental risk of death when added to the clinical model (chi-square 133, p = 0.009), exercise test model (chi-square = 203, p = 0.002), and echocardiographic model (chi-square = 216, p = 0.03). A QRS duration > or =105 ms best identified patients at increased risk. In conclusion, QRS duration is associated with an increased risk of death, even after adjustment for clinical factors, exercise capacity, left ventricular function, and exercise-induced myocardial ischemia. PMID- 16214441 TI - Screening for depression in patients with coronary heart disease (data from the Heart and Soul Study). AB - Major depression is associated with adverse outcomes in patients who have coronary heart disease. How best to identify depression in busy cardiology practices is unknown. We compared the test characteristics of 4 depression screening instruments with an interview diagnosis of depression (Diagnostic Interview Schedule) in 1,024 outpatients who had coronary heart disease. Screening instruments were the 10-item Center for Epidemiologic Studies Depression Scale-10, the Patient Health Questionnaire-9, the Patient Health Questionnaire-2, and a simple 2-item instrument that asks (1) "During the past month, have you often been bothered by feeling down, depressed, or hopeless?" and (2) "During the past month, have you often been bothered by little interest or pleasure in doing things?" Of the 1,024 participants, 224 (22%) had major depression based on the Diagnostic Interview Schedule. Areas under the receiver operating characteristic curves were similar for all instruments (range 0.84 to 0.87). In conclusion, a positive response to 1 of the 2 items was 90% sensitive and 69% specific for depression, with a negative likelihood ratio of 0.14. Thus, negative responses to the 2 items effectively ruled out depression. A score > or =10 on the Patient Health Questionnaire-9 was 54% sensitive and 90% specific, with a positive likelihood ratio of 5.4. Thus, a cutpoint > or =10 was virtually diagnostic for depression. PMID- 16214443 TI - Effect of pravastatin on malondialdehyde-modified low-density lipoprotein levels and coronary plaque regression as determined by three-dimensional intravascular ultrasound. AB - We hypothesized that a reduction in atherogenic malondialdehyde-modified low density lipoprotein (MDA-LDL) levels, which may antagonize the action of atheroprotective high-density lipoprotein cholesterol, leads to coronary plaque regression. This study investigated the effects of pravastatin on the serum levels of MDA-LDL and coronary atherosclerosis. In a 6-month prospective study, 75 patients with stable coronary artery disease were randomly assigned to a pravastatin-treatment group (n = 52) or a control group (n = 23). Volumetric analyses were performed in matched coronary artery segments by 3-dimensional intravascular ultrasound. Pravastatin therapy for 6 months resulted in a decrease in coronary plaque volume (14.4%, p <0.0001) and a corresponding reduction in serum MDA-LDL levels (12.7%, p = 0.0001). In the pravastatin treatment group, the percentage of change in plaque volume correlated with changes in the MDA-LDL and high-density lipoprotein cholesterol levels (r = 0.52 and -0.55, respectively, p <0.0001) but not with the changes in any other lipid levels. Multivariate regression analysis revealed that a reduced MDA-LDL level is an independent predictor of plaque regression, as was an increase in high-density lipoprotein cholesterol. In conclusion, these results suggest that the reduction in the MDA LDL levels induced by pravastatin may serve as a novel marker of coronary atherosclerosis regression. PMID- 16214444 TI - Variability in platelet aggregation following sustained aspirin and clopidogrel treatment in patients with coronary heart disease and influence of the 807 C/T polymorphism of the glycoprotein Ia gene. AB - A broad variability in patient response to dual antiplatelet treatment has been described during the first month of treatment. Data on platelet function profiles in patients on dual antiplatelet therapy for a more sustained period are limited. Whether gene sequence variations of the glycoprotein Ia/IIa receptor influence platelet aggregation in these patients is also unknown. The aim of this study was to characterize platelet aggregation profiles in patients on dual antiplatelet treatment (aspirin plus clopidogrel) for >1 month and to assess whether these may be influenced by the C807T polymorphism of the glycoprotein Ia gene. We included 82 patients, who were divided into 2 groups: carriers (CT + TT genotypes; n = 51) and noncarriers (CC genotype; n = 31) of the mutant T allele. Platelet aggregation was assessed using light transmittance aggregometry after stimuli with adenosine diphosphate (20 micromol/L), collagen (6 microg/ml), and epinephrine (20 micromol/L). A significant variability in the distribution of platelet aggregation was observed in the overall study population, as well as in carriers and noncarriers of the T allele. T allele carriers had increased platelet aggregation compared with noncarriers after stimuli with adenosine diphosphate, collagen, and epinephrine (p <0.05 for all platelet aggregation assays). Thus, platelet aggregation varied significantly in patients on long-term dual antiplatelet treatment and was increased in T allele carriers of the 807C/T polymorphism of the glycoprotein Ia gene. These findings may contribute to the increased ischemic risk observed in these patients. PMID- 16214445 TI - Impact of treatment of coronary artery disease with sirolimus-eluting stents on outcomes of diabetic and nondiabetic patients. AB - Patients with diabetes mellitus are at increased risk for repeat interventions and mortality after coronary angioplasty and stenting. The efficacy of sirolimus eluting stents (SESs) to improve the outcomes of these patients is a focus of interest. In the first 1,407 patients treated with SESs at our institution, 492 were diabetic (insulin dependent diabetes mellitus [IDDM], n = 160 and non insulin-dependent DM [NIDDM], n = 332). The in-hospital and 1- and 6-month clinical outcomes were compared with those of 915 patients without DM (non-DM). The baseline characteristics were similar, except for more women, obesity, previous myocardial infarction, coronary artery bypass grafting, and renal insufficiency in the DM group (p <0.001). Compared with non-DM patients, DM patients had higher in-hospital (p <0.05) and 1-month mortality (p = 0.02). IDDM patients had more in-hospital renal failure (p = 0.04) and Q-wave myocardial infarctions (1.6% vs 0%, p = 0.04) compared with NIDDM patients, and higher mortality (3.1% vs 0.8%, p = 0.04) and subacute stent thromboses (2.3% vs 0.5%, p = 0.07) than non-DM patients at 30 days. At 6 months, DM patients had a higher incidence of Q-wave myocardial infarction, target lesion revascularization-major adverse cardiac events, and composite of death and Q-wave myocardial infarction than non-DM patients (6.0% vs 2.7%, p = 0.01). Late outcomes between the IDDM and NIDDM groups were similar. Multivariate analysis showed diabetes and acute renal failure as independent predictors of target lesion revascularization-major adverse cardiac events. In conclusion, our data showed that, despite a reduction in repeat revascularization, coronary intervention with SESs in diabetic patients is limited by higher mortality at 1 month and a higher incidence of Q-wave myocardial infarction and target lesion revascularization-major adverse cardiac events at 6 months compared with non-DM patients. Careful surveillance is required in IDDM patients undergoing SES implantation. PMID- 16214446 TI - Relation of atherosclerotic changes in retinal arteries to the extent of coronary artery disease. AB - The aim of this study was to explore the relation between atherosclerosis of the retinal arteries and the extent and severity of coronary artery disease (CAD). In 109 patients, aged 40 to 80 years, who underwent coronary angiography for suspected CAD, the degree of retinal arterial atherosclerosis (stages 1 to 4 according to Scheie) was determined. The fundus examination was done using direct ophthalmoscopy by an ophthalmologist blinded to the extent of the CAD. The CAD extent was evaluated by Gensini score, and coronary angiograms were analyzed by 2 expert observers who had no knowledge of the patients' retinal artery status. The extent and severity of retinal vessel atherosclerosis correlated strongly with the extent and severity of CAD. Thus, atherosclerotic changes in the retinal arteries may be a predictor of the extent of CAD. PMID- 16214447 TI - Predictors of outcome after sirolimus-eluting stent implantation for complex in stent restenosis. AB - Few data are available on the effectiveness of sirolimus-eluting stent implantation for the treatment of in-stent restenosis, and no data exist about the predictors of outcome after sirolimus-eluting stent implantation for complex in-stent restenosis (diffuse, proliferative, or total occlusion). From April 2002 to May 2004, 136 patients with 161 complex in-stent restenoses underwent sirolimus-eluting stent implantation. At 9 months, 5 patients had died (3 of cardiac and 2 of noncardiac causes), no reinfarctions had occurred, and 11 target vessel revascularization procedures had been performed. The target vessel revascularization rate was 8%, and the in-segment binary restenosis rate was 17%. The predictors of the risk of recurrence were unstable angina as the clinical presentation of in-stent restenosis, an ostial location of the target lesion, lesion length, and sirolimus-eluting stent diameter < or =2.5 mm. PMID- 16214448 TI - Prognostic value of impaired fasting glucose for outcomes of patients with stable angina pectoris treated with percutaneous coronary interventions. AB - The prognostic value of the newly defined impaired fasting glucose (IFG) range (100 to 109 mg/dl) for the outcomes of patients undergoing percutaneous coronary interventions is unknown. We assessed the composite of death or myocardial infarction at 1 year of follow-up in 189 patients with IFG and 801 patients with a normal fasting glucose (<100 mg/dl), all with stable angina. The 1-year cumulative rate of death or myocardial infarction was 10.3% in the IFG group and 4.4% in the normal fasting glucose group (p = 0.002). In the multivariate model, IFG was an independent predictor of the occurrence of death or myocardial infarction (adjusted hazard ratio 2.30, 95% confidence interval 1.29 to 4.08, p = 0.005). An IFG of 100 to 109 mg/dl in patients with stable angina who undergo percutaneous coronary intervention may identify a patient subset with an increased risk of death or myocardial infarction. PMID- 16214449 TI - Peripheral CD34+ cells and the risk of in-stent restenosis in patients with coronary heart disease. AB - In-stent restenosis represents the major limitation of percutaneous coronary revascularization. The underlying neointimal hyperplasia mainly consists of smooth muscle cells (SMCs), which can be derived from bone marrow cells. We hypothesized that changes in the peripheral progenitor cell counts after coronary stenting may predict the development of restenosis. We prospectively studied men with atherosclerotic coronary artery disease who had undergone successful elective stenting of solitary target lesions (n = 17). Peripheral blood samples were drawn at baseline (before stenting) and 1 day after stenting. The CD34+ cell count was determined by flow cytometry. Follow-up quantitative coronary angiography was performed after 8.1 +/- 2.6 months. Except for longer primary lesions in patients with angiographic restenosis, no significant differences in patient and lesion characteristics were seen. The rate of restenosis (75% vs 11%, p = 0.015) and the extent of diameter stenosis at follow-up (56.9 +/- 26.9% vs 26.5 +/- 16.5%, p = 0.012) were higher in patients with a postprocedural increase in CD34+ cells than in those with a decrease in CD34+ cells. Postprocedural CD34+ cell counts were increased in patients with restenosis but decreased in those without restenosis (p = 0.002). A robust correlation was seen between the change in CD34+ cells and late lumen loss (r = 0.65, p <0.005). In a multivariate regression model, the change in CD34+ cells, lesion length, and preprocedural minimal lumen diameter independently predicted for late lumen loss. In conclusion, an increase in circulating CD34+ cells after coronary stenting constitutes an independent risk factor predicting in-stent restenosis and may be suggestive of their involvement in neointimal hyperplasia. PMID- 16214450 TI - Effectiveness of the stent pull-back technique for nonaorto ostial coronary narrowings. AB - Coronary stenting of nonaorto ostial coronary lesions is challenging because of plaque shift into the main vessel, triggering the use of additional stents. Furthermore, inappropriate coverage of the ostium of the side branch increases the risk of restenosis and target vessel revascularization (TVR). To improve the treatment of nonaorto ostial coronary lesions with a novel interventional technique, we tested the hypothesis that inflating a balloon in the main vessel before stenting the side branch (stent pull-back technique) will limit plaque shifting and reduce the use of additional stents. In addition, proper coverage of the side branch ostium may also reduce 8-month TVR. A case-control design with 100 consecutive patients who underwent drug-eluting stent placement was performed; 55 patients were treated with the stent pull-back technique and 45 patients with a conventional stent technique. Procedural success was 100% for the 2 techniques. The use of additional stents was reduced in the stent pull-back group compared with the conventional stent group (2% vs 18%, p = 0.007). A tendency toward lower ostial miss was also observed in the stent pull-back group (4% vs 13%, p = 0.11). The incidence of in-hospital and 30-day cardiac events was similar between the 2 groups. TVR was lower in the stent pull-back group compared with the conventional group (5% vs 20%; p = 0.03). In conclusion, the stent pull back technique improves the percutaneous treatment of nonaorto ostial coronary lesions. The technique is associated with a lower use of additional stents and improved clinical outcome, reducing TVR at 8 months of follow-up. PMID- 16214451 TI - Non-high-density lipoprotein cholesterol versus low-density lipoprotein cholesterol as a risk factor for a first nonfatal myocardial infarction. AB - Low-density lipoprotein (LDL) cholesterol is the primary lipid parameter targeted to prevent myocardial infarction. Alternatively, non-high-density lipoprotein (HDL) cholesterol includes LDL cholesterol and other atherogenic particles but does not require a fasting sample. Non-HDL cholesterol and LDL cholesterol as predictors of first nonfatal myocardial infarction were compared in 303 patients and 297 controls matched for age, gender, and community within the Boston Area Health Study. Patients were white men or women aged <76 years living in the Boston area, without a history of myocardial infarction or angina pectoris, in whom symptoms of confirmed myocardial infarction began during the 24 hours before admission. After multivariate adjustment for coronary risk factors in unmatched analyses, the corresponding odds ratios (ORs) of a first nonfatal myocardial infarction for non-HDL cholesterol in the second, third, and fourth quartiles were 1.83 (95% confidence interval [CI] 1.07 to 3.14), 2.07 (95% CI 1.23 to 3.49), and 2.33 (95% CI 1.39 to 3.90) (p trend <0.01). For LDL cholesterol, the ORs were 1.10 (95% CI 0.67 to 1.81), 0.87 (95% CI 0.52 to 1.46), and 1.45 (95% CI 0.90 to 2.35) (p trend = 0.16). Including HDL cholesterol in the model increased the ORs and strengthened the test for a trend for LDL cholesterol, whereas the ORs were decreased and the test for a trend was weakened for non-HDL cholesterol. In conclusion, given that non-HDL cholesterol accounts for LDL cholesterol plus other atherogenic particles but does not require a fasting sample, this study suggests that non-HDL cholesterol may be at least as useful as LDL cholesterol to initially screen patients for risk of a first nonfatal myocardial infarction. PMID- 16214452 TI - Dietary magnesium intake and risk of cardiovascular disease among women. AB - This study assessed the hypothesis that greater magnesium intake is associated with reduced risk for cardiovascular disease (CVD), including myocardial infarction (MI) and stroke, in a large prospective cohort of women. In 1993, a semi-quantitative food frequency questionnaire was used to assess magnesium intake in 39,876 female health professionals aged 39 to 89 years who had no history of CVD or cancer. During a median of 10 years of follow-up, 1,037 incident cases of CVD were identified, including 280 nonfatal MIs and 368 strokes. After adjustment for age and randomized treatment status, magnesium intake was not significantly associated with risk for incident CVD. Comparing the highest quintile of magnesium intake (median 433 mg/day) with the lowest quintile (median 255 mg/day), the relative risks were 0.87 (95% confidence interval [CI] 0.72 to 1.05, p for trend = 0.24) for total CVD, 0.88 (95% CI 0.70 to 1.12, p for trend = 0.34) for coronary heart disease (CHD), 1.03 (95% CI 0.72 to 1.49, p for trend = 0.96) for nonfatal MI, 1.11 (95% CI 0.61 to 2.00, p for trend = 0.95) for CVD death, and 0.87 (95% CI 0.64 to 1.18, p for trend = 0.55) for total stroke. Additional adjustment for other CVD risk factors did not materially change the observed null associations. In conclusion, the results do not support the hypothesis that magnesium intake reduces the development of CHD, although a modest inverse association with stroke cannot be ruled out. PMID- 16214453 TI - Comparison of efficacy and safety assessment of fluvastatin in patients <65 years versus > or =65 years of age. AB - This pooled analysis of 30 completed clinical trials assessed the efficacy and safety of fluvastatin in patients <65 (n = 8,037) and patients > or =65 years of age (n = 3,717). The results demonstrated that in patients > or =65 years of age, lipid changes with fluvastatin therapy are equivalent to or slightly better than those observed in patients <65 years of age. Treatment with fluvastatin produced a significantly lower incidence of major cardiovascular clinical end points (major adverse cardiac events [MACEs]) and an increase in the time to a first MACE in the older population. The incidence of adverse events, particularly those of concern with statin therapy, was similar between the placebo- and fluvastatin treated patients and between the different age groups. In conclusion, data derived from the pooled analysis with fluvastatin demonstrate that cardiovascular events are reduced in older high-risk patients to a greater extent compared with younger patients. Furthermore, this pooled analysis supports the use of fluvastatin to lower cholesterol levels in older high-risk patients. PMID- 16214454 TI - Comparison and relation of indirect and direct dynamic indexes of cardiac pumping capacity in chronic heart failure. AB - Cardiac power output (CPO), the product of cardiac output (CO) and mean arterial pressure (MAP), is a direct measure of cardiac pumping capability and is strongly indicative of prognosis and exercise ability in patients with chronic heart failure (CHF). However, it is not as easily measured as indirect indicators of cardiac function, such as peak oxygen consumption (VO(2)) or peak "circulatory power" (CircP), the product of VO(2) and MAP. The relation between direct and indirect indexes of cardiac pumping capacity was evaluated in 219 ambulatory patients with CHF. CircP was found to have a direct and consistent relation with CPO, overall (R = 0.93, p <0.0001) and at peak exercise (R = 0.84, p < 0.0001). The results suggest CircP to be an adequate measure of cardiac pumping capacity when the more directly measured CPO is not available. PMID- 16214455 TI - Usefulness of three-dimensionally guided assessment of mitral stenosis using matrix-array ultrasound. AB - Two-dimensional (2-D) planimetry is limited by the technical demands, time, and observer variability required to locate the minimal orifice area, limiting the confident clinical reporting of mitral valve area (MVA). In 27 consecutive patients, MVA was determined independently by 2 observers using the conventional 2-D method and a new 3-D-guided method. Using a matrix-array probe, the valve was visualized in a long-axis view and a cursor steered to intersect the leaflet tips and provide a perpendicular short-axis plane viewed side-by-side. Two-dimensional and 3-D-guided methods allowed planimetry in 24 patients. Consistent with better orifice localization, 3-D guidance eliminated the overestimation of internal orifice diameters in the planimetered short-axis view relative to the limiting diameter defined by the long-axis view (for 3-D guidance, 0.73 +/- 0.20 vs 0.73 +/- 0.21 cm, p = 0.98, vs 0.90 +/- 0.27 cm in the 2-D short-axis view, p <0.01). Accordingly, mean values for the smallest orifice area by 3-D guidance were less than by 2-D imaging (1.4 +/- 0.5 vs 1.5 +/- 0.5 cm(2), p <0.01), changing the clinical severity classification in 11 of 24 patients (46%). The 2-D method also overestimated MVA relative to 3-D guidance compared with Doppler pressure halftime and (n = 6) Gorlin areas. Phantom studies verified no differences in resolution for the 2 acquisition modes. Three-dimensional guidance reduced intraobserver variability from 9.8% to 3.8% (SEE 0.14 to 0.06 cm(2), p <0.01) and interobserver variability from 10.6% to 6.1% (SEE 0.15 to 0.09 cm(2), p <0.02). In conclusion, matrix-array technology provides a feasible and highly reproducible direct 3-D-guided method for measuring the limiting mitral orifice area. PMID- 16214456 TI - Ventricular remodeling in children with left ventricular dysfunction secondary to various cardiomyopathies. AB - The ventricular remodeling that occurs in adults with left ventricular (LV) dysfunction is characterized by a change in LV shape from an ellipse to more of a sphere and is associated with increased functional mitral regurgitation (MR), decreased exercise tolerance, and poor outcome. There are limited data on the occurrence and importance of LV remodeling in children with LV dysfunction. The purposes of this study were to evaluate in children with LV dysfunction (1) LV shape, (2) changes in LV shape as LV function improves, and (3) the relation between LV shape and functional MR. Children with LV dysfunction were identified and compared with controls. Patient demographics, treatment, and outcomes were noted. Echocardiograms were reviewed for LV function and shape (sphericity index). The echos of a subset of children whose LV ejection fractions (LVEFs) increased by >10% over a follow-up of >6 months were analyzed for changes in sphericity, mitral annulus size, and the degree of MR. Twenty-five children with LV dysfunction were compared with 37 age-matched controls. LV remodeling to a more spherical shape was inversely related to LVEF but was not associated with clinical outcome. In a subset of patients (n = 17) whose LVEFs improved, the reversal of remodeling (decrease in sphericity index) was associated with a decrease in MR and mitral annulus size. In conclusion, LV remodeling occurs in children with LV dysfunction. The reversal of this remodeling with a decrease in the degree of MR and the size of the mitral annulus occurs as systolic function improves. PMID- 16214457 TI - Carotid artery remodeling in middle-aged women with the metabolic syndrome (from the "Progetto ATENA" study). AB - The prevalence of the metabolic syndrome (MS) is rapidly growing, especially in women. This analysis was designed to evaluate whether preclinical vascular changes are detectable in middle-aged women with the MS. Five thousand sixty-two women participated in a prospective study of the causes of cardiovascular disease in women (the "Progetto Atena" study). Three hundred ten women underwent high resolution B-mode ultrasound examinations. Common carotid intima-media thickness (IMT) and internal and external diameters were measured using a computerized system. The National Cholesterol Education Program Adult Treatment Panel III criteria were used to classify participants as having the MS. Seventy-three subjects fulfilled the criteria for the MS. Women with the MS were slightly older and had greater serum total cholesterol and higher levels of serum insulin and lipid peroxidation compared with women without the MS. The group with the MS had a greater mean IMT (1.24 +/- 0.27 vs 1.10 +/- 0.20 mm, p <0.0001) and larger mean values for diameters (7.9 +/- 1.0 vs 7.4 +/- 0.7 mm, p <0.0001 for external diameter; 6.1 +/- 0.8 vs 5.8 +/- 0.6 mm, p <0.005 for internal diameter). The average increase in wall thickness was greater than in vessel diameter, as evidenced by the greater relative wall thickness in women with the MS (0.41 +/- 0.09 vs 0.38 +/- 0.07, p <0.005). In conclusion, carotid structural changes are noninvasively detectable in asymptomatic middle-aged women with the MS. These modifications indicate a dynamic process of arterial remodeling by which the vascular system responds to atherosclerotic stimuli. PMID- 16214458 TI - Ethnic differences and heritability of heart rate variability in African- and European American youth. AB - This study investigated whether heart rate variability (HRV) in young African Americans differed from that in young European Americans. It further examined the genetic and/or environmental sources of HRV variance and to what extent they depend on ethnicity or gender in young twins. Subjects were available from 1 data set including 166 subjects (mean age 16 +/- 2 years; 63 African-Americans) and another including 219 twins (11 singletons [4 African-Americans] and 104 pairs [42 African-Americans]; mean age 15 +/- 2 years). HRV was measured over 256 RR intervals in a supine position. Two time-domain variables, the SD of normal RR intervals (SDNN) and the root-mean-square of successive differences (RMSSD) of normal RR intervals, and 3 frequency-domain variables, high-frequency (HF) power, low-frequency (LF) power, and the LF power/HF power ratio, were used. African Americans had higher RMSSDs (p <0.01) and HF power (p = 0.047) and lower LF power/HF power ratios (p <0.01) than European Americans. These differences remained significant after adjusting for covariates. All HRV parameters were heritable; estimated heritability ranged from 32% to 71%. Model fitting showed no ethnic or gender differences for any measure. SDNN, RMSSD, and HF power were strongly correlated (r values >0.8). One factor explaining >90% of the variance for all 3 measures was identified. The heritability of this combined HRV score was 70%. In conclusion, this study suggests that ethnic differences in HRV already exist in youth, with African-Americans having greater HRV than European Americans. High heritability estimates for HRV measures were observed, and no differences in HRV heritability estimates were noted for ethnicity or gender. PMID- 16214459 TI - Relation of tissue displacement and strain to invasively determined right ventricular stroke volume. AB - The role of echocardiography in the clinical assessment of right ventricular (RV) systolic function remains limited. Limited data exist on the potential use of newer techniques for RV function assessment. Conventional echocardiography and tissue Doppler echocardiography (TDE) were performed during right-sided cardiac catheterization in 46 patients. Thermodilution or the Fick-derived RV stroke volume indexed (RVSVI) indexed to body surface area was used as the reference standard. Univariate and multivariate regression analyses were used to test correlations between RVSVI and various echocardiographic and TDE-derived parameters. In a subset of 12 subjects, changes in echocardiographic and TDE variables to reduced afterload from intravenous epoprostenol were measured. TDE derived RV tissue displacement and systolic strain best predicted the RVSVI (r = 0.63, p = 0.001; r = 0.48, p = 0.002, respectively). The prediction improved after adjustment for tricuspid regurgitation jet vena contracta width (r = 0.74, p < 0.0001; r = 0.60, p < 0.001, respectively). Assuming a RVSVI of > or =30 ml/m(2) as normal, a RV displacement cutoff of 15 mm yielded a sensitivity of 100% and a specificity of 41% for RV dysfunction, and an RV systolic strain cutoff of 20% yielded a sensitivity of 91% and a specificity of 63%. The percentage change of RV systolic displacement correlated well with the percentage change of RVSVI after epoprostenol infusion (r = 0.75, p < 0.001). In conclusion, TDE-derived RV displacement and strain closely correlate with RVSVI and appropriately track load-related changes in RV function. These new parameters may help provide the noninvasive, quantitative assessment of RV function. PMID- 16214460 TI - Scapulothoracic dissociation: evaluation and management. AB - Scapulothoracic dissociation is an infrequent injury with potentially devastating outcomes. Knowledge of this injury is based on small patient series and case reports. The aim of this article is to review the evaluation, management and functional outcomes following scapulothoracic dissociation. Often caused by high traction forces applied to the shoulder girdle, there is a complete loss of the scapulothoracic articulation with lateral scapular displacement and intact skin. This is frequently associated with muscular, ligamentous and osseous injuries to the shoulder girdle, vascular injuries to the subclavian, or axillary, vessels and brachial plexus lesions. In the acute setting, the timely diagnosis of the associated neurovascular injuries is crucial. Severe neurovascular and soft tissue compromise often requires an early above-elbow amputation. Further, complete brachial plexus avulsions are associated with a limited potential for functional recovery. PMID- 16214461 TI - Availability of orthopaedic trauma lists in the United Kingdom. AB - National guidelines recommend that hospitals dealing with acute orthopaedic trauma should have daily, consultant led trauma lists, performed in dedicated trauma theatres. This study examined the availability and organisation of orthopaedic trauma lists in the United Kingdom using a postal survey. Of 228 responding hospitals, only 29% had daily trauma lists. In 17%, elective cases were cancelled to accommodate trauma whilst up to 20% of hospitals had a high proportion of their trauma lists not being consultant led. Twenty-one percent of responding hospitals did not have a dedicated trauma theatre. Our results suggest that most units have a long way to go in meeting national standards regarding the availability of orthopaedic trauma lists. PMID- 16214462 TI - Modification of the Constant-Murley shoulder score-introduction of the individual relative Constant score Individual shoulder assessment. AB - The Constant-Murley shoulder assessment score has proven to be a valuable diagnostic instrument. Thus, in the literature it has been mentioned that the clinical accuracy of this score varies especially when comparing patients in larger, inhomogeneous patient groups. The "relative Constant score" (CS(rel)) tries to minimize these problems by using reference parameters out of healthy age and gender related control groups. The authors of this study tried to show that it is even more accurate to use the functional performance of the uninjured collateral shoulder of the same individual as reference, introducing the "individual relative Constant score" (CS(indiv)). The CS(indiv) and the CS(rel) were compared for 125 consecutive patients with shoulder disorders, and a group of 125 healthy volunteers as a control group. In a non-parametric comparison of the reciever operating characteristics the CS(indiv) shows the higher ability to discriminate between patients and healthy volunteers (p=0.004). This indicates that the individual relative Constant score gives a more accurate view about the functional result for shoulder disorders. It is expected to be more reliable for larger and incoherent patient populations, because specific interindividual differences, regarding the patient's age, gender and constitution are eliminated as well as other individual physiological parameters. PMID- 16214463 TI - Painless reduction of anterior shoulder dislocation by Kocher's method. AB - Kocher described his method of reduction of anterior shoulder dislocation in German, which was misinterpreted. A new element of traction was added which causes considerable pain. We present our experience with the original method in reducing 12 anterior shoulder dislocations without using sedation or anaesthesia. The original method is gentle, painless and requires neither sedation nor anaesthesia. Patient reassurance and confidence is essential. The patient initiates the movements, the surgeon just guiding him through the manoeuvre. It was successful in 10 dislocations. In two dislocations, the humeral head had to be gently guided laterally and superiorly into the glenoid using an index finger in the axilla. The method is easily reproducible after a demonstration. Patients can go home within a few minutes of the procedure. PMID- 16214464 TI - Missed Monteggia injuries. AB - The classical Monteggia injury comprises a dislocation of the radial head with an associated fracture of the ulna. In the variant type, there is no ulnar fracture merely plastic deformation. We performed a retrospective study of all Monteggia injuries from 1992 to 2001. A total of 39 were reviewed, of which 8 were missed (1 classical and 7 variant). Of those, five were male and three female with a mean age of 6.3 years. The mean follow-up was 2.5 years, with a mean interval between injury and diagnosis of 33.5 weeks. Two Monteggia injuries diagnosed within 4 weeks were successfully treated by closed manipulation. The other six required ulnar osteotomy, repair of the annular ligament and stabilisation of the radial head with a transcapitellar pin. A protocol for the diagnosis of Monteggia injuries is described. Doubtful cases require an immediate review since early treatment improves the outcome. Acceptable clinical and radiological results in late diagnosed Monteggia injuries can result from ulnar osteotomy, open reduction of the radial head with repair of the annular ligament and transcapitellar pin stabilisation of the reduced radial head. PMID- 16214465 TI - The use of elastic intramedullary nailing in the stabilisation of paediatric fractures. AB - The treatment of paediatric long-bone fractures remains controversial. Elastic intramedullary nailing has been proposed as an alternative for the treatment of paediatric long-bone fractures. PATIENTS: We treated 68 children in a time span of 9 years with 32 fractures of the forearm, 27 fractures of the femur and nine fractures of the tibia. The mean age was 8.3 years and the mean admission time 2.6 days. RESULTS: Mean consolidation time was 7.9 weeks for the forearm fractures, 11.9 weeks for the femur fractures and 10.6 weeks for the tibial fractures. Full weight bearing for the latter two fracture types was allowed within the first week. There were no major complications. The complications encountered were three hydrops of the knee, four low-grade infections and one delayed union. Leg length discrepancy was only seen in five patients (18%) and was less than 2 cm. DISCUSSION: In femur fractures, we let the parents decide between skeletal traction and intramedullary rods. When confronted with the possible complications (operation-related complications and infection) compared to the advantages (early weight bearing and short admission time), they almost always choose the operative approach. In our opinion, elastic intramedullary nailing is an excellent treatment option for diaphyseal fractures in children with skeletal immaturity, especially of the femur. PMID- 16214466 TI - Early proximal row carpectomy after severe carpal trauma. AB - Complex fracture dislocations of the wrist often result in post-traumatic arthrosis. In these cases, patients can experience severe pain and loss of function of the wrist and as a result many of them end up having a total arthrodesis. In trying to avoid the disadvantages of a total arthrodesis, alternative treatment strategies have been investigated, amongst which proximal row carpectomy (PRC). Basic conditions for a good outcome of PRC are an intact cartilage of the lunate fossa of the distal radius and an intact surface of the head of the capitate for the new radiocapitate joint (, schematic drawing post PRC). Also, an intact (volar) radioscaphocapitate (RSC) ligament is necessary because it plays an essential role in stabilizing the new joint and preventing volar dislocation and ulnar translocation of the distal carpal row. Acute post traumatic PRC can be indicated, but is rarely reported in literature. In this article, we present four patients whom we have treated with early PRC after severe trauma of the wrist. Three patients had a good outcome. In the patient with the bad outcome, the before-mentioned prerequisites were not met, which is discussed. PMID- 16214467 TI - Anterior interosseus nerve injury following tension band wiring of the olecranon. PMID- 16214468 TI - The use of traction to simplify intramedullary fixation of paediatric forearm fractures. AB - A minority of paediatric and early adolescent diaphyseal forearm fractures require operative fixation. The method of closed reduction and intramedullary stabilisation of such fractures using stainless steel pins or flexible titanium nails has well documented success rates. The literature proposing their use advises the wires are inserted with the forearm resting free on a radiolucent arm table. In our experience, this technique is demanding; the fracture sites are unstable, often with soft tissue interposition, and the proximal radial fragment is difficult to control by closed means because of the overlying muscle mass. The technique, we describe, employs a standard Maquet Orthostar traction table (Maquet, Colima Ave, Hylton Riverside, Sunderland, UK) to effect a stable closed reduction of diaphyseal forearm fractures to simplify the procedure of passing the radial wire. The traction is then released to allow elbow flexion and the ulna wire to be passed, the ulna fracture having been reduced by the traction and stabilised by the radial wire. In summary, the use of traction to reduce and stabilise the radial fracture has simplified the surgery and reduced operative time. PMID- 16214469 TI - Origami in dynamic hip screw surgery. PMID- 16214470 TI - Review of the vascularisation of the human Achilles tendon. AB - A region of avascularity mid-way along the length of the Achilles tendon has long been associated with rupture. Whilst it is agreed that this region is the location most common to rupture, the exact vascular distribution appears unclear. Regions of avascularity identified within the tendon have included the origin and insertion, as well as the midsection. This review aims to analyse critically and summarise all previous studies of the vascularisation of the healthy human Achilles tendon, in order to determine the most likely region of avascularity and, thereby establish whether a relationship exists between vascularisation and rupture. Whilst no definitive conclusion was reached, it was concluded that the vascularisation does affect the tensile strength and so rupture vulnerability of the healthy Achilles tendon, although it is unlikely to be either the sole, or most significant, contributor. Other factors, such as thinning and twisting of the tendon at the midsection are mechanical influences that will increase the incidence of rupture by increasing the concentration of stress. PMID- 16214471 TI - Secular trends in rates of unintentional injury deaths among adult Finns. AB - This study assessed the current secular trends in unintentional injury deaths among persons 15 years of age or older in Finland. For this purpose, we obtained from the Finnish Official Cause-of-Death Statistics (OCDS) the data for persons aged 15 years or older whose deaths occurred in 1971--2003 due to an unintentional injury. Among Finnish men, the most drastic change occurred in road traffic crashes. The age-standardised death rate (per 100,000 person-years) of men's road traffic crashes was 47 in 1971 but only 11 in 2003. In contrast, the rate of fall-induced deaths among Finnish men gradually rose, from 18 in 1971 to 24 in 2003, by which time falls became the leading category of men's unintentional injury death. Also alcohol poisonings seemed to bypass road traffic crashes as the cause of men's injury death. Among Finnish women, the age standardised rate of fall-induced deaths decreased till 1975, after which the curve was rather flat. The death rate (per 100,000 person-years) of women's falls was 30 in 1971 and 18 in 2003. During the entire period of 1971--2003, traffic caused fewer deaths in women than men, but the declining trend in women's death rates was also clear, from 17 in 1971 to 5 in 2003. Other unintentional causes accounted for few deaths in women during 1971--2003, although it was of interest that the rate of women's deaths due to alcohol poisoning rose from 1 in 1971 to 4 in 2003. In conclusion, during 1971--2003 falls replaced road traffic crashes as the leading cause of unintentional injury death in adult Finns. The rise in men's fall-induced deaths occurred even with a rate that cannot be explained merely by demographic changes, and therefore, systematic fall prevention measures are needed to control the development. The rising incidence of women's deaths due to alcohol poisoning needs close follow-up. PMID- 16214472 TI - The evolution of an integrated State Trauma System in Victoria, Australia. AB - The incidence of major trauma and associated fatalities in the State of Victoria, Australia, have declined over 20 years following the successful implementation of strategies to modify environmental and behavioural factors that contribute to motor vehicle injuries. However, several system deficiencies in the management of major trauma patients had remained unresolved. To investigate these shortfalls the State Government of Victoria established a task force in 1997 to review trauma and emergency services. The task force adopted the principle of "the right patient to the right hospital in the shortest time" and in 2000 began to deploy an integrated State Trauma System. Implementation of such a system required the designation of specific hospitals of various levels to care for trauma patients; the concentration of trauma expertise at these centres; integration and coordination between the service providers; development of agreed triage and transfer protocols and improved education, training and research programs. A statewide major trauma database was established to enable system monitoring and facilitate further enhancements. The Victorian experience with the development of an integrated trauma system should aid in the development of similar systems nationally and internationally and is described in this paper. PMID- 16214473 TI - Is trauma centre care helpful for less severely injured patients? AB - BACKGROUND: Trauma centres have been shown to reduce the number of preventable deaths from serious injuries. This is due largely to the rapid response of surgeons and health care teams to resuscitate, evaluate, and operate if necessary. Less is known about the effectiveness of trauma centre care on those patients who have not incurred immediate life-threatening problems and may not be as critically injured. The purpose of this study was to review the use of physician and hospital resources for this patient population to determine whether trauma team and trauma centre care is helpful or even needed. METHODS: This was a retrospective study of consecutive trauma patients (n=1592) admitted from 1998 to 2002 to the trauma service of an urban level I trauma centre and recorded in the hospital trauma registry. Patients were triaged in a tiered response to more or less severely injured. All patients' care was directed by trauma surgeons. RESULTS: Of the 1592 patients, 398 (25%) received a full trauma team response (Class I), 1194 were less seriously injured (Class II). The ISS for the Class I patients was 19+/-18 and for Class II patients 10+/-10. Nineteen percent of Class II patients had an ISS>15. Overall mortality in Class II patients was 2% including 20 unexpected deaths. Four hundred and three Class II patients (34%) had multisystem injuries. Of the Class II patients 423 (35%) were sent to the ICU or OR from the ED, 106 of whom required an immediate operation and 345 required an operation prior to discharge. Complications developed in 129 patients (11%), the majority of which were pulmonary. CONCLUSIONS: A large proportion of those patients thought initially to be less severely injured required resources available in a trauma centre, including specialty care, intensive care, and operating room accessibility. Over one-third of these patients had multisystem injuries and almost 20% were considered major trauma, needing prioritisation of care and expertise ideally found in a trauma centre environment. Complications developed in a sizable number of patients. This patient population, because of its heterogeneity and propensity for critical illness, deserves the resources of a trauma centre. PMID- 16214474 TI - Is paramedic judgement useful in prehospital trauma triage? AB - Precise prehospital trauma triage criteria are critical for ensuring patients with severe injuries are transported to trauma centres. Most prehospital trauma triage criteria adopt a combination of physiological, anatomic and mechanism of injury components, but this approach still fails to identify a number of patients with severe injuries and often burdens trauma centres with patients suffering minor injuries. Paramedic judgement has been identified as an alternative method for the triage of trauma patients. This study critically reviewed the literature regarding the ability of paramedics to predict injury severity, and found there is no clear evidence supporting paramedic judgement as an accurate triage method. However, the studies were limited due to significant data losses, variable definitions of major trauma, differences across EMS and trauma care systems, variable paramedic experience levels and incomparable methods of data collection. The role of paramedic judgement in identifying patients with severe blunt anatomic injuries requires further investigation. PMID- 16214475 TI - Anticoagulation management in hip fracture patients on warfarin. AB - The management of patients admitted with a fracture requiring surgery who are taking warfarin anticoagulation is unclear. We examined the anticoagulation management for 33 hip fracture patients on warfarin at the time of admission. Hospital course and complications were recorded on all patients. The mean INR on admission was 3.2 and prior to surgery 2.2. Eight patients (24%) had percutaneous cancellous screws for an intracapsular fracture regardless of the admission INR. In 21 (64%) patients, surgery was delayed whilst the INR came down, with an average delay of 72 h from admission to surgery. No specific treatment to lower the INR, other than wait and watch policy adopted in 11 (33%) of these patients. Pharmacological methods used to reduce the INR were fresh frozen plasma in nine cases, and intravenous Vitamin K in four patients. One patient died from post operative haematemesis and three died from medical complications unrelated to the warfarin therapy. There were no wound haematomas or other bleeding complications. Delaying surgery whilst waiting for the INR to fall to acceptable levels may result in significant delays to surgery and we would recommend a more aggressive policy to enable earlier surgery. PMID- 16214476 TI - The acute medical care costs of fall-related injuries among the U.S. older adults. AB - OBJECTIVES: Falls in the older adults are a major public health concern. The growing population of adults 65 years or older, advances in medical care and changes in the costs of care motivated our study of the acute health care costs of fall-related injuries among the older adults in the United States of America. DESIGN AND SETTINGS: The Market Scan Medicare Supplemental database 1998 was used to estimate reimbursed costs for hospital, emergency department (ED), and outpatient clinic treatments for unintentional falls among older adults. RESULTS: A fall on the same level due to slipping, tripping, or stumbling was the most common mechanism of injury (28%). Mean hospitalisation cost was 17,483 US dollars(S.D.: 22,426 US dollars) in 2004 US dollars. Femur fracture was the most expensive type of injury (18,638 US dollars, S.D.: 19,990 US dollars). The mean reimbursement cost of an ED visit was 236 US dollars and 412 US dollars for an outpatient clinic visit. CONCLUSION: The magnitude of the economic and social costs of falls in older adults underscores the need for active research in the field of falls prevention. PMID- 16214477 TI - Percutaneous compression plating versus compression hip screw fixation for the treatment of intertrochanteric hip fractures. AB - Percutaneous compression plate (PCCP) devices are used for the fixation of intertrochanteric hip fractures by a minimally invasive technique. One hundred and eight patients who underwent this procedure were retrospectively compared with 155 patients who underwent compression hip screw (CHS) fixation. The general characteristics of the two groups, including age, sex, side of injury and co morbidities assessed by the ASA score were similar. The operative time was significantly shorter in the PCCP group (67 versus 87 min, p=0.00). Postoperative blood transfusions were not required in 40% of the patients in the PCCP group compared to 24% of the patients in the CHS group (p<0.01). The rate of systemic postoperative complications was lower in the PCCP group (p=0.02) both in univariate and multivariate analyses. A considerable reduction was observed in cardiovascular complications (OR=3.1, p<0.05). Length of hospitalisation, implant failure and mortality rates were not significantly different between the two study groups. We conclude that the PCCP device offers several advantages over CHS device and may improve the current treatment of intertrochanteric hip fractures while maintaining a similar success rate in fracture fixation. PMID- 16214478 TI - Two-hole side-plate DHS in the treatment of intertrochanteric fracture: results and complications. AB - We reviewed 83 intertrochanteric fractures fixed with a 135 degree dynamic hip screw (DHS) and two-hole side-plate. The mean age was 72 years. There were 13 A1.1, 24 A1.2, 16 A2.1, 29 A2.2 and 1 A2.3 fractures. Established osteoporosis was seen in 55 patients. The mean time to union was 14.5 weeks. Sixty-eight percent had minimal collapse, 24% moderate and 8% severe. Over 80% of moderate and severe collapses were associated with osteoporosis and an unstable fracture pattern. There were four failures: two from lag screw cut-out and two from pull out of the side-plate. PMID- 16214479 TI - Femoral neck-shaft angle in extra-capsular proximal femoral fracture fixation; does it make a TAD of difference? AB - The effect of femoral neck-shaft angle and implant type on the accuracy of lag screw placement in extra-capsular proximal femoral fracture fixation was investigated. Radiographs of all extra-capsular proximal femoral fractures seen in one unit over 18 months were reviewed. Of 399 cases, 307 (237 female, 70 male) were included in the study as they had no contra-lateral proximal femoral metal work. Femoral neck-shaft angle (NSA) of the uninjured hip and magnification adjusted tip-apex distance (TAD) of femoral head lag screw were measured. Type of fixation implant was 135 degrees classic hip screw (CHS) (n=144) or 130 degrees intra-medullary hip screw (IMHS) (n=163). Mean contra-lateral NSA was 130.2 degrees (112.9--148 degrees ) and 64 patients (58 female, 6 male) had a NSA <125 degrees . Mean adjusted TAD was 18.7 mm (5.8--43.8mm) and 88.9% of cases had a TAD of less than 25 mm. TAD values were significantly greater using an IMHS if NSA was <125 degrees than if NSA was >125 degrees (p=0.028). This was not the case with the CHS. The use of the 130 degrees -IMHS in patients with a NSA <125 degrees leads to poorer lag screw placement than if NSA >125 degrees and caution is advocated when using this device in such cases. PMID- 16214480 TI - Pure bilateral sacroiliac dislocation with intact anterior pelvis. PMID- 16214481 TI - Effect of oscillating fluid shear on solute transport in cortical bone. AB - The consequences of an oscillatory fluid shear mechanism on nutrient transport in bone during physical activity and ultrasonic therapy are discussed. During movement, periodic stress on bone creates transient pressure gradients that circulate interstitial fluid through calcified bone. A transport model derived from oscillatory Taylor-Aris dispersion phenomena was used to predict a ratio of effective-to-molecular diffusivity, K/D, for solutes of varying sizes up to 50 nm in diameter, in pores filled with interstitial fluid and pericellular matrix. The magnitude of the estimated transport enhancement depended on the molecular size, pore dimension, applied frequency and the displacement of the fluid during pressurization. For oscillation frequencies and amplitudes corresponding to those experienced during normal human activity, transport enhancements of up to 100 fold are expected for molecules larger than 5 nm in diameter. Enhancements of up to one order of magnitude, due to ultrasound stimulations in the MHz frequency range, are also expected for 7-nm-sized solutes. No effects are anticipated for ions, whose molecular diffusion time is too fast relative to the oscillation frequency. This model is expected to be useful for understanding differences in bone growth as a function of type of movement or to develop new physical therapies. PMID- 16214482 TI - Length-tension properties of ankle muscles in chronic human spinal cord injury. AB - Contracture, or loss of range of motion (ROM) of a joint, is a common clinical problem in individuals with spinal cord injury (SCI). In order to measure the possible contribution of changes in muscle length to the loss of ankle ROM, the active force vs. angle curves for the tibialis anterior (TA) and gastrocnemiussoleus (GS) were measured in 20 participants, 10 with SCI, and 10 gender and age matched, neurologically intact (NI) individuals. Electrical stimuli were applied to the TA and GS motor nerves at incremented angles of the entire ROM of the ankle and the resulting ankle and knee torques were measured using a multi-axis load cell. The muscle forces of the TA and GS were calculated from the torque measurements using estimates of their respective moment arms and the resulting forces were plotted against joint angle. The force-angle relation for the GS at the ankle (GSA) was significantly shifted into plantar flexion in SCI subjects, compared to NI controls (t-test, p<0.001). Similar results were obtained based upon the GS knee (GSK) force-angle measurements (p<0.05). Conversely, no significant shift in the force-angle relation was found for the TA (p=0.138). Differences in the passive ROM were consistent with the force-angle changes. The ROM in the dorsiflexion direction was significantly smaller in SCI subjects compared to NI controls (p<0.05) while the plantar flexion ROM was not significantly different (p=0.114). Based upon these results, we concluded that muscle shortening is an important component of contracture in SCI. PMID- 16214483 TI - Regional material property alterations in porcine femoral arteries with atheroma development. AB - We have developed a novel methodology that permits assessment of regional vascular mechanical property alterations in the presence of atheroma in vivo employing a Yucatan miniswine model with induced lesions. Femoral arteries were imaged with intravascular ultrasound. Image data were segmented and, following three-dimensional reconstruction, underwent finite element and sensitivity analysis with optimization to identify regions with altered vascular mechanical properties. All regions were compared to histological analysis. In 12 animals with 8 weeks of endothelial cell denudation and high cholesterol diet (induced atherosclerosis), the elastic modulus initially decreased with early lesion development and then increased with increasing fibrosis-(elastic modulus-all values x10(4)Pa-mean+/-SEM) histologically normal (non-denuded control segment) elements 9.73+/-0.01, fatty elements 9.53+/-0.01, fibrofatty elements 9.41+/ 0.03, and fibrous elements 9.68+/-0.02 (all p<0.001 vs. normal elements). Wall thickness, however, increased with atheroma formation. These data demonstrate decreasing vascular material properties with early lesions, followed by an increase as lesions progress. This methodology permits determination of areas with early atheroma development, follow atheroma progression, and potentially evaluate interventions aimed at decreasing atheroma load and normalizing vascular material properties. PMID- 16214484 TI - Finite element simulation of early creep and wear in total hip arthroplasty. AB - Polyethylene wear particulate has been implicated in osteolytic lesion development and may lead to implant loosening and revision surgery. Wear in total hip arthroplasty is frequently estimated from patient radiographs by measurement of penetration of the femoral head into the polyethylene liner. Penetration, however, is multi-factorial, and includes components of wear and deformation due to creep. From a clinical perspective, it is of great interest to separate these elements to better evaluate true wear rates in vivo. Thus, the aim of this study was to determine polyethylene creep and wear penetration and volumetric wear during simulated gait loading conditions for variables of head size, liner thickness, and head-liner clearance. A finite element model of hip replacement articulation was developed, and creep and wear simulation was performed to 1 million gait cycles. Creep of the liner occurred quickly and increased the predicted contact areas by up to 56%, subsequently reducing contact pressures by up to 41%. Greater creep penetration was found with smaller heads, thicker liners, and larger clearance. The least volumetric wear but the most linear penetration was found with the smallest head size. Although polyethylene thickness increases from 4 to 16 mm produced only slight increases in volumetric wear and modest effects on total penetration, the fraction of creep in total penetration varied with thickness from 10% to over 50%. With thicker liners and smaller heads, creep will comprise a significant fraction of early penetration. These results will aid an understanding of the complex interaction of creep and wear. PMID- 16214485 TI - Finite element analysis of a new customized composite post system for endodontically treated teeth. AB - This paper investigated the mechanical behavior of a new customized post system built up with a composite framework presently utilized for crowns, bridges, veneers and inlay/onlay dental restorations. The material has been shaped so to follow perfectly the profile of the root canal in order to take advantage of the better mechanical properties of composites with respect to metallic alloys commonly used for cast posts. The analysis has been carried out with 3D finite element models previously validated on the basis of experimental work. The new post system has been compared to a variety of restorations using either prefabricated or cast posts. The structural efficiency of the new restoration has been evaluated for an upper incisor under different loading conditions (mastication, bruxism, impact). Results prove that maximum stress values in restored teeth are rather insensitive to post types and materials. However, the new customized composite restoration allows to reduce significantly the stresses inside the dentinal regions where conservative clinical interventions are not possible. PMID- 16214486 TI - Lubrication regimes in mesothelial sliding. AB - To function normally, the lungs, heart, and other organs must undergo changes in shape and size, sliding against surrounding body walls. It is not known whether the delicate mesothelial surfaces covering these organs and body wall are in contact during sliding, or if hydrodynamic pressure in the lubricating liquid increases separation between their surfaces. To address this question, we measured the coefficient of friction (mu) of the mesothelial surface of nine rat abdominal walls sliding in saline on a smooth glass surface. Sliding at physiological velocities of 0.0123-6.14 cm/s with normal stresses of 50-200 Pa, mu varied with velocity (P<0.001). On average, mu was relatively high at low speeds (0.078 at 0.041 cm/s), decreased to a minimum at intermediate speeds (0.034 at 1.23 cm/s), and increased slightly again at higher speeds (0.045 at 6.14 cm/s), consistent with a mixed lubrication regime in which there is at least partial hydrodynamic separation of surfaces. We conclude that mesothelial surfaces, sliding under physiological conditions, are protected from excessive shear by hydrodynamic pressures that increase separation of surfaces. PMID- 16214487 TI - Four-dimensional model of the lower extremity after total hip arthroplasty. AB - We have developed a four-dimensional (4D) model of the lower extremities after total hip arthroplasty in patients. The model can aid in preventing complications such as dislocation and wearing of the sliding surface. The skeletal structure and implant alignment were obtained from CT data. We applied registration method using CAD data to estimate accurate implant alignment from scattered CT data. The reconstructed three-dimensional (3D) skeletal model was combined with motion capture data that were acquired by an optical tracking system. We displayed the patient's skeletal movement and analyzed several parameters that relate to complications. The patient's skeletal model was superimposed onto video footage that was taken by a synchronized and calibrated digital video camera. For validation of the measurement error in this system, we used open MRI to evaluate the relative movement between skin markers and bones. This system visually represents not only the 3D anatomical structure, but also 4D dynamic functions that include the time sequential transitions of components and their positions. The open MRI results indicated that the average error in hip angle was within 5 degrees for each static posture. This system enables clinicians to analyze patient's motions on the basis of individual differences. We found that our system was an effective tool in providing precise guidance of daily postoperative motions that was individualized for each patient. This system will be applicable for surgical planning, assessment of postoperative activities, and the development of new surgical techniques, materials, and prosthetic designs. PMID- 16214488 TI - Strain distribution in an elastic substrate vibrated in a bioreactor for vocal fold tissue engineering. AB - A bioreactor previously described was used to quantify the shear strain along a bioengineered tissue scaffold driven at low audio frequencies (20-200 Hz). Standing wave patterns were calculated analytically by solving a classical boundary value problem for a vibrating string under tension and bending stiffness. Boundary conditions were non-traditional in that small pivot arms at the endpoints allowed neither the displacement nor the velocity to go to zero. The calculations were corroborated with stroboscopic measurement of the motion of the material in the bioreactor. Results indicate that shear strains up to 0.2 can be obtained at low frequencies (20 Hz), with a gradual decrease at higher frequencies due to the decaying amplitude response of the mechanical driver. The bioreactor may be useful for approximating the Young's modulus of the material in situ by probing for resonance frequencies in the standing wave pattern. A yet unsolved problem is a variable drag coefficient along the length of the material due to fluid turbulence in the culture medium. PMID- 16214489 TI - Influence of patella alta on knee extensor mechanics. AB - The purpose of this study was to compare the knee extensor mechanics in persons with and without patella alta. Thirteen subjects with patella alta and 14 subjects with normal patellar position participated in the study. Sagittal and axial MR images of the knee were acquired at 0 degrees , 20 degrees , 40 degrees , and 60 degrees of knee flexion. Measurements of actual moment arm, patellar ligament/quadriceps tendon force ratio, quadriceps effective moment arm, and joint reaction force/quadriceps force ratio were obtained. There were no differences between groups in terms of actual moment arm. However, subjects with patella alta had significantly larger patellar ligament/quadriceps tendon force ratios (1.04+/-0.02 vs. 0.92+/-0.02) and quadriceps effective moment arms (4.40+/ 0.09 vs. 4.00+/-0.09 cm) when averaged across the range of knee flexion angles tested. There was no difference in the joint reaction force/quadriceps force ratio between groups. The observed differences in knee extensor mechanics suggest that individuals with patella alta have a more efficient knee extensor mechanism and would be expected to generate similar joint reaction forces per unit quadriceps force compared to subjects with normal patellar position. Therefore, persons with patella alta may experience less patellofemoral joint reaction force to overcome the same knee flexion moment in the range of 0 degrees -60 degrees of knee flexion. PMID- 16214490 TI - The damping properties of the venous plexus of the heel region of the foot during simulated heelstrike. AB - The damping mechanisms that are operational in the heel pad during the impact phase of locomotion have the important function to protect the musculo-skeletal system from injuries. How this is achieved is still not fully understood, as is for instance illustrated by the 'heel pad paradox', the observation that in vivo and in vitro experiments yielded widely different results. This paradox could so far only partially be explained. In the light of this paradox, and a previous study by our group, we conjectured that the venous plexus might contribute as a hydraulic shock absorber to the damping properties of the heel pad. To investigate this hypothesis in vivo, heel pads of 11 volunteers were subjected to pendulum impact tests, using velocities of 0.2, 0.4, and 0.6 m/s, and three physiologically different, consecutive conditions: (i) a relatively empty venous plexus, (ii) a congested venous plexus, and (iii) a decongested venous plexus. At congestion, the maximum impact force decreased slightly but significantly by 2.6% at 0.2 m/s and 1.8% at 0.4 m/s. This effect was no longer found at 0.6 m/s. Although these effects are rather small, they confirm the fundamental hypothesis that the venous plexus contributes to the damping properties of the heel pad during walking. It is likely that some underestimation of the effect has occurred. PMID- 16214491 TI - Combined finite-element and rigid-body analysis of human jaw joint dynamics. AB - The jaw joint plays a crucial role in human mastication. It acts as a guidance for jaw movements and as a fulcrum for force generation. The joint is subjected to loading which causes tensions and deformations in its cartilaginous structures. These are assumed to be a major determinant for development, maintenance and also degeneration of the joint. To analyze the distribution of tensions and deformations in the cartilaginous structures of the jaw joint during jaw movement, a dynamical model of the human masticatory system has been constructed. Its movements are controlled by muscle activation. The articular cartilage layers and articular disc were included as finite-element (FE) models. As this combination of rigid-body and FE modeling had not been applied to musculoskeletal systems yet, its benefits and limitations were assessed by simulating both unloaded and loaded jaw movements. It was demonstrated that joint loads increase with muscle activation, irrespective of the external loads. With increasing joint load, the size of the stressed area of the articular surfaces was enlarged, whereas the peak stresses were much less affected. The results suggest that the articular disc enables distribution of local contact stresses over a much wider area of the very incongruent articular surfaces by transforming compressive principal stress into shear stress. PMID- 16214492 TI - Trabecular bone fracture healing simulation with finite element analysis and fuzzy logic. AB - Trabecular bone fractures heal through intramembraneous ossification. This process differs from diaphyseal fracture healing in that the trabecular marrow provides a rich vascular supply to the healing bone, there is very little callus formation, woven bone forms directly without a cartilage intermediary, and the woven bone is remodelled to form trabecular bone. Previous studies have used numerical methods to simulate diaphyseal fracture healing or bone remodelling, however not trabecular fracture healing, which involves both tissue differentiation and trabecular formation. The objective of this study was to determine if intramembraneous bone formation and remodelling during trabecular bone fracture healing could be simulated using the same mechanobiological principles as those proposed for diaphyseal fracture healing. Using finite element analysis and the fuzzy logic for diaphyseal healing, the model simulated formation of woven bone in the fracture gap and subsequent remodelling of the bone to form trabecular bone. We also demonstrated that the trabecular structure is dependent on the applied loading conditions. A single model that can simulate bone healing and remodelling may prove to be a useful tool in predicting musculoskeletal tissue differentiation in different vascular and mechanical environments. PMID- 16214493 TI - Spatial fiber type distribution in normal human muscle Histochemical and tensiomyographical evaluation. AB - The variability of fiber type distribution in nine limb muscles was examined with histochemical and tensiomyographical (TMG) methods in two groups of 15 men aged between 17 and 40 years. The aim of this study was to determine the extent to which the relative occurrence of different fiber types and subtypes varies within human limb muscles in function to depth and to predict fiber type proportions with a non-invasive TMG method. The distribution of different fiber types varied within the muscles, as a function of depth, with a predominance of type 2b fibers at the surface and type 1 fibers in deeper regions of the muscle. For all the analyzed muscles the contraction times measured at stimulus intensity 10% of supramaximal stimulus (10% MS) were significantly (p<0.05) shorter than the contraction times measured at 50% of supramaximal stimulus intensity (50% MS). The Pearson's correlation coefficient between percentage of type 1 muscle fibers measured at the surface of the muscle and contraction time at 10% MS, obtained by TMG was statistically significant (r=0.76,P<0.01). Also the Pearson's correlation coefficient between percentage of type 1 muscle fibers measured in the deep region of the muscle and contraction time at 50% MS obtained by TMG was also statistically significant (r=0.90,P<0.001). These findings suggest that the contraction time obtained by TMG may be useful for non-invasive examining of muscle fiber types spatial distribution in humans. PMID- 16214494 TI - Structural effects of an innovative surgical technique to repair heart valve defects. AB - The structural and functional effects of the "edge-to-edge" technique on the human mitral valve have been investigated, paying particular attention to the diastolic phase. An advanced finite element model of the valve has been developed, using a hyperelastic material schematization, suitable geometry and constraint conditions, and an effective fluidodynamic analysis. The edge-to-edge suture has been applied on this model and the diastolic phase has been simulated. The results of this calculation show that the operation increases the transvalvular pressure and the maximum stress in the leaflets, which reaches a level similar to that of the systolic phase. The influence of suture position and extension, and the mitral annulus dimension has also been investigated. The results indicate that a lateral location of the stitch is better than a central one, both regarding valve functionality (pressure level and mobility) and internal stresses level, that a longer suture worsens the valve functionality but reduces the stresses level, finally, that the dilatation of the mitral annulus does not affect the valve functionality but increases the stresses level. PMID- 16214495 TI - Adaptive system identification applied to the biomechanical response of the human trunk during sudden loading. AB - Epidemiological evidence indicates that sudden loading of the torso is a risk factor for low back injury. Accurately quantifying the time-varying loading of the spine during sudden loading events and how these loading profiles are affected by workplace factors such as fatigue, expectation, and training can potentially lead to intervention strategies that can reduce these risks. Electromyographic and trunk motion data were collected from six male participants who performed a series of sudden loading trials with varying levels of expectation (no preview, 300-ms audible preview), fatigue (no fatiguing exertion preceding sudden load, short duration/high intensity fatiguing exertion preceding sudden load), and training (untrained, trained). These data were used as inputs to an adaptive system identification model wherein time-varying lower back stiffness, torque, work, and impulse magnitudes were calculated. Results indicated that expectation significantly increased peak and average stiffness by 70% and 113%, respectively, and significantly decreased peak torque, work, and impulse magnitudes by 36%, 50%, and 45%, respectively. Training significantly decreased peak torque and work by 25% and 34%, respectively. The results also showed a significant interaction between expectation and training wherein training had a positive effect during the trials with preview but no effect during the trials with no preview (increased peak stiffness by 17% and decreased impulse magnitude by 43%). PMID- 16214496 TI - Passive motion characteristics of the talocrural and the subtalar joint by dual Euler angles. AB - The objective of this study is to validate previous descriptions of hindfoot kinematics using dual Euler angle methods in a passive cadaveric model. The dual Euler angle method was chosen so as to facilitate description of the translational and rotational movement occurring at both the ankle and subtalar joints. A non-metal experimental set-up was fabricated to generate motion in foot cadaver specimens. Three-dimensional kinematic data of the ankle joint complex was collected from ten knee-below foot cadaver specimens using a 'Flock of Birds' electromagnetic tracking device. The data correlates well with previously published kinematic descriptions of the ankle subtalar joint complex. Both the ankle and subtalar joint show 6 degree of freedom motion and multiaxial characteristics. The motions of the talocrural joint, the talocalcaneal joint, and the gross motion between the foot and the shank were analyzed. During dorsiflexion-plantarflexion the motion of the calcaneus with respect to the tibia occurs mainly at the ankle joint, with little motion at the subtalar joint. The subtalar joint contributes more than the ankle joint during inversion-eversion. PMID- 16214497 TI - The effects of orientation, temperature, and displacement magnitude changes on the sonometrics system accuracy. AB - The accuracy and reliability of a sonomicrometry system (Sonometrics Corporation, Ontario, Canada) was evaluated for its potential use in measuring 3-D in vivo joint kinematics. Distances between different sets of piezoelectric crystals were measured through a salt solution using ultrasound technology. We evaluated crystal-to-crystal distance under simulated in vivo conditions of changing crystal orientation and displacement magnitude. Crystal-to-crystal distance was also evaluated under changing solution temperature, since the crystals may be used at different temperatures. The 2 mm round and peg crystals were accurate to within 0.5mm for 0 through 180 degrees rotations, but the 2mm round suture loop crystals were only reliable at 0 degrees rotation. The speed of sound through a salt solution (and hence the distance between crystals) versus temperature was fit using a second order polynomial, C=1421.1+3.9808T-3.09x10(-2)T2, with an R2 value of 0.9998. The translational error was less than 0.072 mm for crystal displacements of 0.012, 0.2, 1.0, and 5.0 mm. The system was also accurate under dynamic conditions with translational errors that were less than 0.045 mm under 0.65 Hz motion. These results suggest that the Sonometrics crystals possess attributes (translational accuracy and rotational independence) that could provide the basis for a system capable of measuring joint kinematics. PMID- 16214498 TI - Methodology to study intimal failure mechanics in human internal carotid arteries. AB - While the incidence of blunt carotid artery injuries is low, the mortality rate is extremely high (40%). Clinical evidence indicates that the intimal region of the artery often sustains failure, while maintaining the integrity of the outer layers. This condition may lead to delayed ischemic symptoms, commonly reported in clinical literature. To date, the mechanical properties of the intima relative to the outer vessel layers have not been quantified in the human carotid artery. The purpose of the present study was to develop a methodology to determine the longitudinal mechanical properties of the human internal carotid artery in tension, with an emphasis on intimal failure. This was accomplished by opening the vessel at the mid-diameter level, creating an 'I'-shaped testing specimen, subjecting the specimen to failure loading, documenting the stretch characteristics of the intimal and adventitial sides in the temporal domain, and correlating the synchronized videography with mechanical loading. Intimal failure data were quantified using stress and strain parameters in conjunction with digital videography of the intimal and adventitial sides. The present methodology can be used to determine the mechanical properties of the intima relative to ultimate carotid artery failure. These data will assist in the understanding of blunt carotid artery injuries, its diagnosis and treatment. PMID- 16214499 TI - Using numerical approximation as an intermediate step in analytical derivations: some observations from biomechanics. AB - We present four examples to illustrate the use of a type of numerical approximation as an intermediate step in analytical derivation of seemingly complicated biomechanical equations. The method involves examination of curve shapes to elucidate useful underlying trends, which may otherwise be overlooked through consideration of only the equations themselves. Two examples of the method's use are drawn from recently published results in the area of experimental methods in biomechanics of very soft tissues, and two others are taken from our current work on cartilage tissue mechanics. We think that such observations provide a useful means of circumventing complexity issues when deriving models for biomechanical analysis, and further that the method, while simple in concept, could be effective in a range of biomechanics applications. PMID- 16214500 TI - Vowels, syllables, and letter names: differences between young children's spelling in English and Portuguese. AB - Young Portuguese-speaking children have been reported to produce more vowel- and syllable-oriented spellings than have English speakers. To investigate the extent and source of such differences, we analyzed children's vocabulary and found that Portuguese words have more vowel letter names and a higher vowel-consonant ratio than do English words. In a spelling experiment, we found that Portuguese speakers used more vowels, but did not produce more syllabic spellings, than did English speakers. The differences that we observed are attributable to quantitative differences in the languages and their writing and letter name systems. They do not support the widespread idea that speakers of Romance languages pass through an additional, syllabic, stage of development. PMID- 16214501 TI - Stabilization of white blood cells and immunologic markers for extended analysis using flow cytometry. AB - We evaluated whole blood samples drawn from 25 healthy donors and 20 HIV-infected donors into K3EDTA and Cyto-Chex BCT blood collection tubes for CD4, CD8, and CD3 cell counts (HIV Panel). Samples collected in Cyto-Chex BCT were stored at room temperature and tested by 4-color flow cytometry at 6 h, 3 days, and 7 days after isolation for CD4, CD8, and CD3 absolute cell counts/microl and compared with samples collected in K3EDTA tubes and tested at 6 h. Regardless of donor type, the samples collected in Cyto-Chex BCT and tested on day 7 yielded results that were statistically indistinguishable (with correlation coefficients of 0.96 or greater) from samples collected in K3EDTA tubes and tested at 6 h. We conclude that whole blood samples collected in Cyto-Chex BCT are stabilized for their marker phenotype for at least 7 days after phlebotomy. PMID- 16214502 TI - Main pulmonary artery to innominate artery shunt during hybrid palliation of hypoplastic left heart syndrome. PMID- 16214503 TI - Extensive atrial wall resection in a patient with symptomatic idiopathic right atrial enlargement. PMID- 16214504 TI - For everything there is a season. PMID- 16214505 TI - Optimizing cardiac cell therapy: from processing to delivery. PMID- 16214506 TI - The impact of stage and cell type on the prognosis of pulmonary neuroendocrine tumors. AB - OBJECTIVE: In pulmonary neuroendocrine tumors the realization that the extent of nodal disease is related to cell type has led to a controversy as to which is the dominant prognostic factor, stage or morphology. To clarify this relationship, we studied patients treated at our institution over a 23-year period. METHODS: This is a historical cohort study of patients with confirmed pulmonary neuroendocrine tumors who underwent lung resection from 1980 through 2003. Survivors were contacted by telephone, and recurrences were confirmed by means of histopathology. Cox proportional hazards regression was used to ascertain the joint influence of several risk factors on survival. RESULTS: The mean age of the cohort was 54 years (standard deviation, 15 years), and 100 (57%) were men. The cell types for the 177 eligible patients were typical carcinoid in 89 (50%), atypical carcinoid in 15 (8%), large cell in 22 (13%), and small cell in 51 (29%). The median time to follow-up was 7 years (first to third quartile, 2-12 years), and overall 5- and 10-year survivals were 86% (79%-90%) and 81% (74% 87%), respectively. The univariable predictors of survival were age (P = .001), nodal stage (P = .01), and cell type (P < .001). In the final multivariable model only age (P = .04) and cell type (P < .001) remained as independent predictors. The hazard of death among patients with large cell or small cell lung cancer was highest in the first year and a half after diagnosis, reducing drastically thereafter. CONCLUSIONS: In pulmonary neuroendocrine tumors cell type is the predominant determinant of survival. The survival of patients with each cell type is sufficiently diverse to warrant different management strategies. Conservative resection is feasible for typical carcinoids, but the effects of adjuvant chemotherapy need to be evaluated for the other subgroups. PMID- 16214507 TI - Progress toward an ambulatory pump-lung. AB - OBJECTIVES: Currently available therapies for acute and chronic lung diseases have not been effective and have various problems associated with the technologies used. We present a novel active mixing pump-lung with the goal of providing total respiratory support to ambulatory patients. METHODS: The pump lung is based on the concept of active mixing oxygenation within a constrained vortex. The rotation of hollow-fiber membranes disrupts the concentration boundary layer, increasing gas exchange efficiency, and simultaneously pumps the blood. Consequently, the amount of membranes required to achieve gas transfer sufficient for total respiratory support is considerably small. A series of studies, including computational design, experimental bench testing, and in vivo animal experiments, have been performed to implement this concept into a viable artificial pump-lung device. RESULTS: A series of pump-lung prototypes with a membrane surface area of 0.17 to 0.5 m2 were designed and characterized in vitro with bovine blood, demonstrating extremely high gas exchange efficiency. The prototype with a gas exchange surface area of 0.5 m2 was evaluated in calves. The device provided oxygen transfer of approximately 115 mL/min for respiratory support of an animal for up to 5 days. CONCLUSIONS: Progress to date suggests a high likelihood of success for an extracorporeal shorter-term lung that can be switched in and out like dialysis devices. Our device is unique in that it incorporates an integrated pumping and active mixing principle for excellent gas transfer and eliminates the need of the native right ventricle's ability to power blood through the artificial and natural lungs. PMID- 16214508 TI - The significance of estrogen receptor beta in 301 surgically treated non-small cell lung cancers. AB - OBJECTIVE: Estrogen receptor expression in lung cancer has been understudied, particularly in light of its potential biologic importance in the epidemic of lung cancer in women. The expression of estrogen receptors was investigated to better understand the possible role of sex hormones in lung cancer. METHODS: A total of 301 patients with surgically resected non-small cell lung cancers of stages I to III were explored. Sections of paraffin-embedded tumor samples were stained with estrogen receptor alpha and estrogen receptor beta antibodies. Tumors with moderate-to-strong nuclear staining in at least 50% of the tumor cells were scored as positive for overexpression. RESULTS: The overall frequency of overexpression for estrogen receptor beta was 45.8% (138/301). It was detected most frequently in female patients (in 54.3% of 127 tumors vs 39.7% of 174 tumors in men, P = .012). However, there was no estrogen receptor alpha nuclear staining detectable in non-small cell lung cancers. Interestingly, a significant correlation between estrogen receptor beta expression, stage of disease, grade of differentiation, smoking status, vascular invasion, and survival in patients with stage II and III disease was found. By using multivariate analysis of survival among patients with stage II and III disease, estrogen receptor beta overexpression, stage II tumor, well differentiation, nonsmoking status, and lack of vascular invasion were significantly favorable prognostic factors. CONCLUSIONS: The results presented here show for the first time that immunohistochemical expression of estrogen receptor beta can be used as a prognostic indicator in patients with surgically resected stage II and III non small cell lung cancers. These observations might offer a possibility for hormonal therapy in patients with lung cancer. PMID- 16214509 TI - Intercostal muscle flap reduces the pain of thoracotomy: a prospective randomized trial. AB - BACKGROUND: Thoracotomy is associated with significant pain and morbidity. METHODS: We performed a prospective randomized trial over 4 months. Patients were randomized to a standard posterior-lateral thoracotomy or an identical procedure, except an intercostal muscle was harvested from the lower rib (to protect the intercostal nerve) before chest retraction. To ensure an equal distribution among both groups, patients were stratified by race, sex, and type of pulmonary resection. All patients received similar pain management. Pain was assessed by using multiple pain scores during hospitalization and after discharge. Outcomes assessed were pain scores, spirometric values, analgesic use, and activity level. RESULTS: There were 114 patients. The median time for intercostal muscle harvesting was 3.7 minutes. The numeric pain scores were lower for the intercostal muscle group on postoperative days 1 and 2 and at weeks 1, 2, 3, 4, 8, and 12 (P < .05 for all). In addition, patients in the intercostal muscle group had a smaller decrease in spirometric values, were less likely to be using analgesics, and were more likely to have returned to normal activity. CONCLUSIONS: The harvesting of an intercostal muscle flap before chest retraction decreases the pain of thoracotomy and leads to a lower decrease in spirometry. In addition, patients have less pain at 1, 2, 3, 4, 8, and 12 weeks postoperatively and are less likely to be using narcotics. Finally, it offers a pedicled muscle flap that takes little time to harvest and is able to buttress all bronchi after lobectomy. PMID- 16214510 TI - Carpentier-Edwards supra-annular aortic porcine bioprosthesis: clinical performance over 20 years. AB - OBJECTIVE: Experience with the Carpentier-Edwards supra-annular porcine bioprosthesis (Edwards Lifesciences, Irvine, Calif) has been evaluated longitudinally over 20 years. Clinical performance was evaluated by actuarial and actual analysis. Hemodynamic performance was evaluated by echocardiographic/Doppler assessment. Morphology of structural failure was evaluated from pathologic examinations. METHODS: From 1981 through 1999, 1823 patients (mean age, 68.9 +/- 10.9 years; range, 19-89 years) underwent 1847 procedures. Concomitant coronary artery bypass was performed in 788 (42.7%) patients. Previous valve procedures were performed in 107 (5.8%) patients, and other cardiac procedures were performed in 87 (4.7%) patients. RESULTS: The overall valve-related complication rate was 4.36% per patient-year (630 patients), with a fatality rate of 0.96% per patient-year (139 patients). Patient survival at 18 years was 15.8% +/- 1.6%. Overall late mortality rate was 6.3% per patient-year. Overall actual cumulative freedom at 18 years from reoperation was 85.0% +/- 1.2%, valve-related mortality was 88.7% +/- 1.1%, and valve-related residual morbidity was 96.3% +/- 5.0%. Actual freedom from structural valve deterioration at 18 years was 86.4% +/- 1.2% overall, 90.5% +/- 1.8% for age 61 to 70 years, and 98.2% +/- 0.6% for age greater than 70 years. Structural valve deterioration presented with pathologic evidence consistent with stenosis in 27.6% and insufficiency in 72.4%. Hemodynamic performance at 1 year revealed normal effective orifice area indexes for sizes 23 to 27 mm and mild-to-moderate reduction for size 21 mm. CONCLUSIONS: The Carpentier-Edwards supra-annular aortic porcine bioprosthesis continues to provide excellent freedom from structural valve deterioration and overall freedom from valve-related residual morbidity, mortality, and reoperation up to 18 years. Hemodynamic performance is satisfactory. The prosthesis remains recommended for patients older than 70 years and for patients 61 to 70 years of age, especially when comorbid risk factors are not anticipated to provide extended survival. PMID- 16214511 TI - Correlation of autologous skeletal myoblast survival with changes in left ventricular remodeling in dilated ischemic heart failure. AB - OBJECTIVES: The effect of autologous skeletal myoblast transplantation has not been rigorously studied in the setting of end-stage ischemic heart failure free of concomitant coronary revascularization. The aims of the present study were to determine autologous skeletal myoblast survival and its effects on left ventricular function and remodeling in sheep with dilated ischemic heart failure. METHODS: Ischemic heart failure (left ventricular ejection fraction, 30% +/- 2%; left ventricular end-systolic volume index, 82 +/- 9 mL/m2) was created in sheep (n = 11) with serial left circumflex coronary artery microembolizations. Instruments were inserted for the long-term determination of left ventricular global and regional dimensions, hemodynamics, and pressure-volume analysis after autologous skeletal myoblast transplantation (approximately 3.0 x 10(8) myoblasts; heart failure plus autologous skeletal myoblast group, n = 5) or without (heart failure-control group, n = 6). Measurements were performed in conscious animals. RESULTS: Autologous skeletal myoblast-derived skeletal muscle was found in all injected animals at 6 weeks. In ischemic heart failure, autologous skeletal myoblast cardiomyoplasty failed to improve systolic (left ventricular ejection fraction, 29% +/- 4%; dP/dT(max), 2863 +/- 152 mm Hg/s; end systolic elastance, 1.6 +/- 0.22) or diastolic (left ventricular end-diastolic pressure, 21 +/- 2 mm Hg; time constant of relaxation (Tau), 34 +/- 4 ms; dP/dT(min), -1880 +/- 68 mm Hg/s) function. There was, however, attenuation in the left ventricular dilatation after autologous skeletal myoblast transplantation (change in end-systolic volume index, 14% +/- 4% vs 32% +/- 6%; P < .05). The effects of autologous skeletal myoblast-derived skeletal muscle were exclusive to the left ventricular short-axis dimension and dependent on autologous skeletal myoblast survival (R2 = 0.59, P = .006, n = 11). CONCLUSIONS: Autologous skeletal cardiomyoplasty was able to attenuate left ventricular remodeling in sheep with end-stage ischemic heart failure. PMID- 16214512 TI - Aortic root replacement with a novel decellularized cryopreserved aortic homograft: postoperative immunoreactivity and early results. AB - OBJECTIVE: Novel antigen-reduction SynerGraft technology (CryoLife Inc, Kennesaw, Ga) reduces the cellular components of a cadaver homograft with an enzymatic and washing process, leaving the extracellular matrix intact. We report the immunoreactivity (positive panel reactive antibody assay) and early operative results of the CryoLife SynerGraft aortic valve conduit homograft. METHODS: Twenty-two patients (age 53 +/- 14 years, range 31-80 years) from April 2002 to July 2003 underwent aortic root replacement with a CryoLife SynerGraft aortic valve conduit homograft (CryoLife Inc, Kennesaw, Ga) for congenital or acquired aortic valve disease, aortic aneurysm with aortic valve disease, or native or prosthetic aortic valve endocarditis. Baseline percentage positive panel reactive antibody results were negative (<10%) for all and were assessed at 1 month, 3 months, and 1-year. Homograft function was evaluated by echocardiography. RESULTS: Early mortality was 0%. Two late deaths at 1 postoperative year were unrelated to homograft function. At 1 postoperative month, panel reactive antibody results were negative in 20 patients (91%). At 3 months, 19 of 22 patients (86%) had negative results, including 1 with previous positive results. At 1 year, 19 of 20 patients (95%) had negative results, including 2 of the 3 with previous positive results. The mean aortic valve gradients were 12 +/- 8 mm Hg (n = 21) at discharge and 11 +/- 7 mm Hg (n = 18) at 1 year. At a mean follow up of 30.3 +/- 5.2 months, the mean gradient was 8.8 +/- 6.3 mm Hg. CONCLUSION: The SynerGraft decellularization technology successfully removed antigens from an aortic valve homograft conduit. Aortic root reconstruction with the CryoLife SynerGraft aortic valve homograft resulted in low transvalvular gradients, similar to those seen with standard cryopreserved homografts. These early results suggest an advancement in homograft technology. The low panel reactive antibody response may enhance durability by eliminating immune complex-mediated reaction against the homograft. The lack of allosensitization in patients who may require organ transplantation in the future is an added benefit. PMID- 16214513 TI - In vitro evaluation of aortic valve prosthesis in a novel valved conduit with pseudosinuses of Valsalva. AB - OBJECTIVE: This study was undertaken to determine whether the presence of vortices immediately above a prosthetic aortic valve could negatively influence the in vitro hydrodynamic performances of a biologic or mechanical valve implanted in a new Dacron polyester fabric conduit that incorporates sculpted sinuses of Valsalva. METHODS: With a computer-controlled pulse duplicator, the in vitro performance (pressure differences, closure and leakage volumes, and energy losses) of a 25-mm mechanical or biologic prosthesis implanted in a standard Dacron straight conduit or in the new Dacron graft with a sculpted sinus were analyzed and compared. RESULTS: The mechanical and biologic prostheses at 7 L/min cardiac output showed pressure drops across the valve of 8.72 mm Hg and 13.45 mm Hg, respectively, when inserted in the new Valsalva-style graft and of 7.97 mm Hg and 12.94 mm Hg, respectively, when inserted in the standard graft. The closure and leakage volumes for mechanical valves were higher than those for biologic valves; however, the presence or absence of sinuses did not result in significant differences in closure and leakage volumes. The maximal total energy losses were 5.89% and 9.49% for mechanical and biologic valves, respectively. No differences were evident between the two different Dacron grafts for each prosthetic heart valve. CONCLUSION: The normal opening and closing behavior of a prosthetic aortic valve was not altered or modified by a different root shape above the heart valve. The presence of vortices inside the pseudosinuses of Valsalva did not influence the hydrodynamic properties of the biologic and mechanical valves tested. PMID- 16214514 TI - Quality of life outcomes after coronary artery bypass graft surgery: relationship to neuropsychologic deficit. AB - OBJECTIVES: Neuropsychologic assessment after coronary artery bypass graft surgery indicates cognitive deficits, but data on their effect on health-related quality of life are relatively sparse. The present study assessed neuropsychologic deficits, self-reported health-related quality of life, and mood, together with proxy rating of patients' activities after bypass surgery. More specifically, the study examines the relationship between these varied outcome measures. METHODS: A prospective longitudinal assessment of 71 male patients was performed over a 6-month period: 1 week before surgery and postoperatively at 2 and 6 months. Within-patient change was assessed with the neuropsychologic test battery and procedures recommended by the Consensus Panel, the Short Form-36 to measure self-reported health-related quality of life, and the Hospital Anxiety and Depression Scale questionnaire to assess anxiety and depression. Proxy ratings were documented with the Functional Activities Questionnaire. RESULTS: Poor preoperative health-related quality of life was largely unrelated to medical variables. Cognitive deficit was found in 42% of patients at 2 months and 22% of patients at 6 months. Physical health-related quality of life improved, but benefit for emotional and social functioning was unconvincing, especially over the short term. Although cognitive deficits were largely unrelated to Short Form-36 health-related quality of life, and only partially related to anxiety and depression, they were associated with proxy ratings of patient functioning. CONCLUSIONS: Physical health-related quality of life benefits were confirmed, but adverse cognitive change and only limited emotional benefit were evident after coronary artery bypass graft surgery. Perceived physical gains are unrelated to any cognitive deficit, but the latter correlated with the emotional status of the patient 6 months after surgery and are of concern to close relatives. These issues should be addressed in patient care. PMID- 16214515 TI - Early and midterm patency of the proximal anastomoses of saphenous vein grafts made with a Symmetry Aortic Connector System. AB - OBJECTIVE: The purpose of this study was to investigate (1) the early and midterm patency rates in saphenous vein grafts that were anastomosed with the Symmetry Aortic Connector System (St Jude Medical, Inc, St Paul, Minn) and (2) risk factors for graft occlusion. METHODS: Thirty-one patients underwent off-pump coronary artery bypass grafting for proximal saphenous vein graft anastomoses with the aortic connector system. Intraoperative graft flow was studied with transit time flowmetry, and angiography was performed before discharge in 29 cases. Midterm (at least 1 year after the operation) saphenous vein graft patency was assessed by coronary angiography or 3-dimensional coronary computed tomography in 27 cases. RESULTS: Postoperative angiography demonstrated a 100% patency rate and no significant stenosis at the connector-anastomosed sites. The 1-year patency rate of the saphenous vein grafts with the aortic connector system was 92.6%, with 2 cases of saphenous vein graft occlusion. Both of these cases had low graft flow and poor left ventricular function documented during the operation. CONCLUSION: Once surgical errors had been ruled out, the 1-year patency rate of proximal saphenous vein grafts anastomosed with the aortic connector system was favorable. Poor left ventricular function and low graft flow during the operation were risk factors for midterm graft occlusion. Selection of the target vessel and its runoff may also be an important determinant of long term patency. PMID- 16214517 TI - Endovascular repair of aneurysm after previous surgical coarctation repair. AB - OBJECTIVES: Late aneurysm formation has been reported after every type of surgical coarctation repair, with rupture of such aneurysms being responsible for approximately 7% of all deaths. Secondary surgical repair carries a significant mortality and morbidity. According to the positive experience with endovascular therapy of atherosclerotic thoracic aortic aneurysms, it is worthwhile to evaluate the concept of minimally invasive endovascular stent grafting for secondary repair of postsurgical aneurysms. METHODS: Data were collected prospectively on consecutive patients who presented with postcoarctation false aneurysms. RESULTS: Since 1999, in a cohort of 46 endovascularly treated patients with thoracic aortic pathologies, 3 patients with postcoarctation false aneurysms underwent endoluminal stent-graft placement. All of these procedures were technically successful without 30-day or 1-year procedure-related mortality. After a mean follow-up of 19 months (range, 7.8-33.5 months), all aneurysm remain excluded without endoleak. CONCLUSIONS: According to the current limited experience of small series and short periods of follow-up, the endoluminal repair seems to be a promising alternative to redo operations for postsurgical thoracic aneurysms associated with coarctation repair. Long-term follow-up is required to assess the durability of the stent-graft treatment. PMID- 16214516 TI - Left ventricular volume and function after endoventricular patch plasty for dyskinetic anteroapical left ventricular aneurysm in sheep. AB - OBJECTIVE: Endoventricular patch plasty (the Dor procedure) has gained favor as a surgical treatment for heart failure associated with large anteroapical myocardial infarction. We tested the hypothesis that the Dor procedure reduces left ventricular volume, increases end-systolic elastance, decreases diastolic compliance, and maintains left ventricular function. METHODS: In 6 male Dorsett sheep, the left anterior descending coronary artery and its second diagonal branch were ligated 40% of the distance from apex to base. Sixteen weeks after myocardial infarction, a Dor procedure was performed with a Dacron patch that was 50% of the infarct neck dimension. Absolute left ventricular volume was measured with magnetic resonance imaging, and left ventricular pressure and relative left ventricular volume changes during pharmacologic preload reduction were measured with a volume conductance catheter 2 weeks before and 2 and 6 weeks after the Dor procedure. End-systolic elastance, diastolic compliance, and Starling relationships were calculated from the resultant left ventricular pressure/volume loops. RESULTS: Two weeks after the Dor procedure, the left ventricular volume at end systole and end diastole was significantly reduced, and there was no redilation at 6 weeks. Six weeks after the Dor procedure, the ejection fraction was significantly increased. Although stroke volume increased slightly at 6 weeks, the change was not significant. The slopes of end-systolic elastance, diastolic compliance, and Starling relationships were unchanged at 2 and 6 weeks. CONCLUSIONS: The Dor procedure significantly reduces left ventricular volume. Unlike linear repair, left ventricular volume changes seem stable. The ejection fraction is improved, and left ventricular function (stroke volume and the Starling relationship) is maintained. PMID- 16214518 TI - In vitro hydrodynamics, cusp-bending deformation, and root distensibility for different types of aortic valve-sparing operations: remodeling, sinus prosthesis, and reimplantation. AB - OBJECTIVE: Preserving aortic valve cusps during operations for aortic root pathology theoretically offers several advantages over alternative prosthetic valve-bearing conduits. Functional properties of different valve-sparing techniques under defined conditions are not well studied. METHODS: Fresh porcine aortic roots were investigated in a pulsatile flow simulator, either native root or after different types of valve-sparing procedures (remodeling, sinus prosthesis, and reimplantation). Functional parameters, such as transvalvular pressure gradient, closing volume, cusp-bending deformation, and distensibility at different levels of the root were analyzed. RESULTS: The mean pressure gradient was highest in reimplantation techniques (8.4 +/- 1.8 mm Hg) compared with sinus prostheses (7.2 +/- 0.9 mm Hg, P = .01) and remodeling techniques (6.8 +/- 1.0 mm Hg, P = .002), mirror imaging the closing volume (reimplantation, 1.5 +/- 0.4 mL; sinus prostheses, 2.3 +/- 0.7 mL [P < .001]; remodeling, 3.4 +/- 1.1 mL [P < .001]). Bending deformation indices increased significantly from remodeling (0.45 +/- 0.05) and sinus prostheses (0.58 +/- 0.06) to reimplantation techniques (0.73 +/- 0.09). Dynamic changes in area of all techniques were decreased at the sinotubular junction and the commissural and sinus levels when compared with those seen in native roots but increased at the annular level for techniques with unfixed annulus (remodeling and modified sinus prosthesis). CONCLUSIONS: In vitro the various aortic valve-sparing operations differed characteristically in their ability to spare valve function, none of them completely meeting native valve behavior. The remodeling techniques exhibited valve dynamics closest to those of the native aortic root. The more the aortic valve is fixed with noncompliant prosthetic material, the more the native root dynamics are impaired. PMID- 16214519 TI - Does a bidirectional Glenn shunt improve the oxygenation of right ventricle dependent coronary circulation in pulmonary atresia with intact ventricular septum? AB - OBJECTIVE: There is a risk of myocardial ischemia in patients with pulmonary atresia and intact ventricular septum associated with the right ventricle dependent coronary circulation. In this patient group, the oxygen delivery to the myocardium depends on the oxygen saturation of the right ventricular cavity. We hypothesized that bidirectional Glenn shunt would improve the oxygenation of right ventricle-dependent coronary circulation relative to a systemic-pulmonary artery shunt. The reduction of systemic venous return to the right atrium due to a bidirectional Glenn shunt could increase the oxygen saturation of the right ventricle in the clinical setting, when the mixture of systemic and pulmonary venous blood is unchanged at the atrial level. METHODS: Patients with right ventricle-dependent coronary circulation were defined as those with right ventricle-coronary artery fistulas plus stenoses of the right or left coronary arteries. For 7 patients with right ventricle-dependent coronary circulation before and after bidirectional Glenn shunt, cardiac catheterization was performed and the oxygen saturation of the right ventricular cavity was measured. RESULTS: For all 7 patients, the bidirectional Glenn shunt was performed at a mean age of 18 months. Ischemic changes in the electrocardiogram before the bidirectional Glenn shunt improved after the procedure in 2 patients. The oxygen saturation of the right ventricular cavity before the bidirectional Glenn shunt was 54.6 +/- 8.8%, and that after the BGS significantly increased to 75.6% +/- 5.8% (P < .01). All 7 patients have subsequently undergone the Fontan procedure with excellent results. CONCLUSION: Early bidirectional Glenn shunt could prevent progression of myocardial ischemia in pulmonary atresia with intact ventricular septum with right ventricle-dependent coronary circulation. PMID- 16214520 TI - Protein oxidation injury occurs during pediatric cardiopulmonary bypass. AB - OBJECTIVE: Proteins are the major effectors of biological structure and function. Oxidation-induced changes to protein structure can critically impair protein function, with important pathologic consequences. This study was undertaken to examine whether oxidation-induced changes to protein structure occur during pediatric cardiopulmonary bypass and to examine the association with postoperative outcome. METHODS: Elevation of the 3,4-dihydroxyphenylalanine content of a protein relative to its native tyrosine content indicates structural damage due to oxidation. Protein 3,4-dihydroxyphenylalanine/native tyrosine ratios were measured before surgery and up to 6 hours after institution of cardiopulmonary bypass in 24 children undergoing repair of congenital heart disease, who were prospectively selected to form a cyanotic and comparable acyanotic control group. Results were correlated with perioperative variables and postoperative outcomes. RESULTS: Elevation of protein 3,4 dihydroxyphenylalanine/tyrosine ratios above baseline (0.48 mmol/mol [SD, 0.11 mmol/mol] vs 0.36 mmol/mol [SD, 0.13 mmol/mol]; P = .001) occurred within 30 minutes of initiating cardiopulmonary bypass in cyanotic but not in acyanotic children and correlated inversely with preoperative arterial oxygen saturation (R = -0.52; P = .03). Protein 3,4-dihydroxyphenylalanine/tyrosine ratios were also increased above baseline at 120 minutes (0.44 mmol/mol [SD, 0.12 mmol/mol]; P = .007) and 180 minutes (0.40 mmol/mol [SD, 0.14 mmol/mol]; P = .01) after the institution of cardiopulmonary bypass in children who underwent prolonged procedures. Elevation of 3,4-dihydroxyphenylalanine/tyrosine during prolonged procedures was associated with postoperative arrhythmias and the need for increased inotropic support (P = .001). CONCLUSIONS: Oxidative injury to proteins occurs during pediatric cardiopulmonary bypass. Cyanotic children are most at risk, particularly those undergoing prolonged procedures, in whom elevation of the protein 3,4-dihydroxyphenylalanine/tyrosine ratio is associated with increased postoperative morbidity. PMID- 16214521 TI - Distribution of cardiac output and oxygen delivery in an acute animal model of single-ventricle physiology. AB - BACKGROUND: When single-ventricle physiology is established acutely (ie, after a Norwood procedure), the combination of limited cardiac output and hypoxemia could result in limited oxygen transport to systemic organs. This study investigates the regional distribution of cardiac output and oxygen delivery after creation of single-ventricle physiology. METHODS: Single-ventricle physiology was created in 8 piglets, and 8 other piglets served as sham control animals. Aortopulmonary shunt, echocardiography-guided atrial septostomy, tricuspid valve avulsion, and pulmonary artery occlusion allowed the left ventricle to support systemic and pulmonary circulations. Physiologic parameters and regional blood flow were determined at baseline and at 30 and 120 minutes after conversion to single ventricle physiology. Parameters were compared by means of 1-way and 2-way analysis of variance. RESULTS: Single-ventricle physiology resulted in lower diastolic arterial pressure, oxygen saturation, and arterial oxygen saturation (P < .05), whereas hemoglobin was unchanged. Cerebral blood flow increased markedly in control animals (P = .04). In contrast, in single-ventricle physiology regional blood flow was unchanged in the brain, higher in the myocardium (P = .1), and mildly reduced in low-priority organs (liver, kidneys, and bowel). Cerebral oxygen delivery increased in control animals, whereas in animals with single-ventricle physiology, oxygen delivery decreased in the brain, liver, kidneys, and bowel (P < .05) and was unchanged in the myocardium. Total-body oxygen delivery decreased in animals with single-ventricle physiology (P < .001) but not in control animals. Total-body oxygen consumption was unchanged in both groups. CONCLUSIONS: This study shows that in acute single-ventricle physiology hypoxemia and limited regional blood flow reduce oxygen transport to low-priority organs and partly to the brain. These findings might contribute to the understanding of gastrointestinal and neurologic complications in children with single-ventricle physiology. PMID- 16214522 TI - Pulmonary atresia with intact ventricular septum: predictors of early and medium term outcome in a population-based study. AB - OBJECTIVES: Pulmonary atresia with intact ventricular septum is a form of congenital heart disease usually associated with right-heart hypoplasia, with considerable morphologic heterogeneity and often poor outlook. Ascertainment of risk factors for poor outcome is an important step if an improvement in outcome is to be achieved. METHODS: The UK and Ireland Collaborative study of Pulmonary Atresia with Intact Ventricular Septum is an ongoing population-based study of all patients born with this disease from 1991 through 1995. All available clinical, morphologic, and investigative variables were directly reviewed, and risk factor analysis was performed for poor outcome. RESULTS: One hundred eighty three patients presented with pulmonary atresia with intact ventricular septum. Fifteen underwent no procedure, and all died. Of the remainder, 67 underwent a right ventricular outflow tract procedure (catheter or surgical), 18 underwent an outflow tract procedure with shunt, and 81 underwent a systemic-to-pulmonary shunt alone. One- and 5-year survival was 70.8% and 63.8%, respectively. Results from Cox proportional hazards model analysis showed that low birth weight (P = .024), unipartite right ventricular morphology (P = .001), and the presence of a dilated right ventricle (P < .001) were independent risk factors for death. The presence of coronary artery fistulae, right ventricular dependence, or the tricuspid valvar z score did not prove to be risk factors for death. After up to 9 years of follow-up, 29% have achieved a biventricular repair, 3% a so-called one-and-a-half ventricular repair, and 10.5% a univentricular repair, with 16.5% still having a mixed circulation (41% died). CONCLUSIONS: This population-based study has shown which features at presentation place an infant in a high-risk group. This is important information for counseling in fetal life and for surgical strategy after birth. PMID- 16214523 TI - Upregulation of hypoxia inducible factor is associated with attenuation of neuronal injury in neonatal piglets undergoing deep hypothermic circulatory arrest. AB - BACKGROUND: Prolonged deep hypothermic circulatory arrest is known to cause neurological injury. Hypoxia inducible factor, a transcription factor that mediates adaptive changes during hypoxia, is neuroprotective in models of ischemic brain injury, in part by upregulating erythropoietin. This study tested the hypothesis that upregulation of hypoxia inducible factor and erythropoietin by preconditioning with hypoxia or the hypoxia-mimetic agents deferoxamine and cobalt chloride would be neuroprotective in a piglet model of deep hypothermic circulatory arrest. METHODS: Anesthetized neonatal piglets were randomized to 4 preconditioning groups (15 per group): hypoxia, deferoxamine, cobalt chloride, or control (NaCl vehicle). Brain hypoxia inducible factor and erythropoietin contents were assessed by means of Western blotting at 3, 8, and 24 hours after treatment (n = 3 per time point). Twenty-four hours after treatment, 6 to 7 animals per group underwent cardiopulmonary bypass and 110 minutes of deep hypothermic circulatory arrest. After recovery, serial neurobehavioral examinations were conducted for 6 days, after which histopathologic brain injury and neuronal apoptosis (cleaved caspase 3) were assessed. RESULTS: Erythropoietin expression was not significantly increased by any of the pretreatment strategies. In contrast, there was a significant upregulation of hypoxia inducible factor by pretreatment with deferoxamine and cobalt chloride (P = .002). Neurobehavioral measures revealed no significant differences in time to recovery or extent of injury. Examination of histopathologic brain injury in the hippocampus revealed that pretreatment with deferoxamine (0.4 +/- 0.3) and cobalt chloride (0.5 +/- 0.3) were associated with significantly less neuronal loss than pretreatment with hypoxia or control (2.8 +/- 0.5, P = .004). Finally, cleaved caspase 3 (a marker of apoptotic cell death) was also shown to be diminished in the cobalt and deferoxamine groups, but the difference was not significantly different from the value in the control group. CONCLUSIONS: In contrast to hypoxia, deferoxamine and cobalt chloride preconditioning upregulated hypoxia inducible factor and were associated with histopathologic neuroprotection after exposure to cardiopulmonary bypass and prolonged deep hypothermic circulatory arrest. PMID- 16214524 TI - Tracheostomy in infants and children after cardiothoracic surgery: indications, associated risk factors, and timing. AB - BACKGROUND: Respiratory insufficiency in children after cardiothoracic surgery delays weaning from the ventilator and prolongs intensive care unit stay. There is little consensus as to the indications for tracheostomy and its safety in this population. METHODS: We reviewed our institutional experience in 37 consecutive infants and children (median age, 8.6 months; weight, 7.2 kg) requiring a tracheostomy after cardiothoracic surgery between January 1998 and December 2001, with follow-up to June 2003. RESULTS: Twenty-four children underwent tracheostomy after corrective (n = 15) or palliative (n = 9) surgery for congenital heart disease, 8 had undergone thoracic transplantation, and 5 had undergone thoracic surgery. Median duration of pretracheostomy ventilation was 30 days, and median total duration of ventilation was 73 days. Tracheostomy was performed earlier in patients undergoing transplantation (median of 20 days postoperatively), with a duration of ventilation of 34 days. No patient experienced mediastinitis, and a wound infection in 1 child was the only identified complication. Twenty-two children survived to hospital discharge, of whom 15 have since been decannulated; 6 still have a tracheostomy in situ and 1 has been lost to follow-up. A number of preoperative and postoperative factors were identified in this cohort. These were preoperative respiratory insufficiency, a history of neonatal ventilation, the need for cardiac reoperations, diaphragmatic paralysis, tracheobronchomalacia, neurological comorbidity, and associated chromosomal abnormalities. CONCLUSION: Tracheostomy can be performed safely and without increased risk of complications in infants and children early after cardiothoracic surgery. The presence of identifiable factors in patients in whom weaning has been unsuccessful should alert clinicians to early consideration of tracheostomy. PMID- 16214525 TI - Systemic venous oxygen saturation after the Norwood procedure and childhood neurodevelopmental outcome. AB - OBJECTIVE: Neonates with hypoplastic left heart syndrome have impaired systemic oxygen delivery and also have a high risk of hypoxic ischemic brain injury with resultant neurodevelopmental impairment. We hypothesized that decreased postoperative oxygen delivery, as measured on the basis of systemic venous oxyhemoglobin saturation, would be related to persistent neurodevelopmental abnormality assessed in childhood. METHODS: Early perioperative hemodynamic data, prospectively acquired from neonates undergoing staged palliation of hypoplastic left heart syndrome by using deep hypothermic circulatory arrest with uniform perioperative management, were tested for relationship to later neurodevelopmental outcome assessed at age 4 years. RESULTS: Complete hemodynamic and neurodevelopmental data were available in 13 patients aged 7 +/- 8 days at the time of the Norwood procedure and aged 4.5 +/- 0.7 years at follow-up assessment. The subjects scored significantly below the population mean for motor, visual-motor integration, and composite neurodevelopmental outcomes. The 5 (38%) patients with abnormal outcomes had significantly lower postoperative systemic venous oxygen saturation values than those with normal outcomes (46% +/- 8% vs 56% +/- 6%, P = .024). Standard hemodynamic parameters did not differentiate patient outcomes. The risk of abnormal outcome increased with increasing time at a systemic venous oxygen saturation of less than 40% (P < .001). A multivariate model of deep hypothermic circulatory arrest time, systemic venous oxygen saturation, blood pressure, and carbon dioxide tension accounted for 79% of the observed variance (P < .001). CONCLUSIONS: Decreased systemic oxygen delivery in the neonatal postoperative period is associated with hypoxic ischemic brain injury and childhood neurodevelopmental abnormality. Measures of systemic oxygen delivery should be used to guide perioperative strategies to reduce the risk of hypoxic-ischemic brain injury. PMID- 16214526 TI - Chronic hypoxemia increases myocardial cytochrome oxidase. AB - OBJECTIVE: Cyanotic patients have potentially decreased tissue oxygen tension. Cytochrome oxidase catalyzes the reduction of oxygen and is integral to adenosine triphosphate production. Cytochrome oxidase subunit I, the active site, is encoded by mitochondrial DNA. Using a newborn swine model of chronic hypoxemia, we evaluated ventricular cytochrome oxidase subunit I mRNA and protein expression and assessed cytochrome oxidase activity. METHODS: Thirty-two newborn piglets underwent thoracotomy and placement of a pulmonary artery-to-left atrium shunt or sham operation. Two weeks later, partial pressure of arterial oxygen, hematocrit, and left ventricular shortening fraction values were compared with baseline values. Northern blot hybridization and protein immunoblotting for ventricular cytochrome oxidase subunit I were performed. Cytochrome oxidase kinetic activity was measured. Heme a,a3 content and turnover number were determined. Significance was assessed with a t test. RESULTS: Baseline partial pressure of arterial oxygen and hematocrit values were similar. Hypoxemic piglets had a lower partial pressure of arterial oxygen of 38 +/- 10 mm Hg (P < .001) and higher hematocrit value of 31.4% +/- 2.9% (P < .001) compared with a partial pressure of arterial oxygen of 140 +/- 47 mm Hg and hematocrit value of 24.6% +/- 3.9% after the sham operation. Baseline and postprocedure left ventricular shortening fraction were similar within and between groups. Chronic hypoxemia increased right ventricular and left ventricular cytochrome oxidase I mRNA and protein by more than 1.4-fold. Cytochrome oxidase activity increased significantly in hypoxemia by 2.5-fold compared with that seen after the sham operation. Heme a,a3 content and turnover number increased by 1.5-fold during hypoxemia. CONCLUSIONS: Chronic hypoxemia increases cytochrome oxidase I message, protein expression, and activity. The increase in kinetics was due to increased enzyme content and catalytic activity. This is a possible adaptive mechanism that might preserve organ function during chronic hypoxemia. PMID- 16214527 TI - Ventricular constraint in dilated cardiomyopathy: a new, compliant textile mesh exerts prophylactic and therapeutic properties. AB - BACKGROUND: Dilated cardiomyopathy is associated with a progressive decrease in cardiac function, leading to end-stage heart failure. We aimed to stop this process by mechanically constraining the heart with a new, compliant textile mesh. METHODS: In 16 male Munich minipigs (50 +/- 7 kg), dilated cardiomyopathy with congestive heart failure was induced through 4 weeks of rapid ventricular pacing (220 beats/min). In the early-mesh group (n = 8), a polyvinylidene fluoride mesh was positioned around both ventricles before pacing was started. In the other group (n = 8), experimental dilated cardiomyopathy through rapid pacing was induced (no mesh). After mesh grafting, rapid pacing was continued (late mesh). RESULTS: Rapid pacing in the no-mesh group (control group) significantly decreased both systolic (cardiac output, peak systolic pressure, and the derivative of pressure increase [dP/dt(max)]) and diastolic (minimum rate of pressure rise [dP/dt(min)] and left ventricular end-diastolic pressure) variables, whereas these variables remained almost unchanged in the early-mesh group. In the late-mesh group the passive-elastic constraint not only prevented further deterioration but even exerted reverse remodeling to some extent (dP/dt(max) and left ventricular end-diastolic pressure, P < .05). CONCLUSIONS: Ventricular constraint with the new mesh seems to be a prophylactic and therapeutic option in cardiac insufficiency caused by ventricular dilation. This passive-elastic cardioplasty induced reverse remodeling of dilated hearts and significantly improved diastolic and systolic ventricular function. PMID- 16214528 TI - Persistence of marrow stromal cells implanted into acutely infarcted myocardium: observations in a xenotransplant model. AB - OBJECTIVE: It has been reported that unmatched adult bone marrow stromal cells could be tolerated by immune-competent allotransplant or xenotransplant recipients under various conditions. This study examined whether xenogeneic bone marrow stromal cells implanted immediately after myocardial infarction can survive and differentiate, attenuating deterioration in left ventricular function. METHODS: In groups I and II (n = 34), myocardial infarctions were created in immunocompetent adult Lewis rats by proximal left coronary artery ligation. In group I, 3 x 10(6)lacZ-labeled mouse bone marrow stromal cells were immediately injected into the peri-infarct area of the left ventricle, whereas in group II, only culture medium was injected. There were 10 early and 4 late deaths. At 4 weeks after injection, hearts were stained for beta-galactosidase and troponin IC. In groups IIIA and IIIB, lacZ-labeled mouse skin fibroblasts were implanted into rat myocardium (n = 10 each) with and without left coronary artery ligation, respectively, and the rats were killed serially. In group IV, animals underwent sham surgery (n = 5, no deaths). At 4 weeks, surviving rats in groups I, II, and IV (n = 10, n = 10, and n = 5, respectively) underwent blinded transthoracic echocardiography for ventricular function studies. RESULTS: In group I, labeled mouse-derived bone marrow stromal cells were found within rat myocardium that stained positively for troponin IC 4 weeks after implantation. Functionally, mean left ventricular ejection fraction (P = .007), stroke volume (P = .03), and fractional shortening (P = .02) were all significantly higher in group I than in group II. In groups IIIA and IIIB, mouse fibroblasts induced cellular infiltration with rapid loss of donor cells. No labeled cells were found after 4 days. In group IV, there was no change in cardiac function. CONCLUSION: Xenogeneic bone marrow stromal cells implanted into acutely ischemic myocardium induced by coronary artery ligation were immunologically tolerated, survived and differentiated, resulting in a cardiac chimera which improved left ventricular function. This unique immunologic tolerance may suggest the feasibility of using bone marrow stromal cells as universal donor cells. PMID- 16214529 TI - Long-term support of 9 patients with the DeBakey VAD for more than 200 days. AB - OBJECTIVE: Pulsatile left ventricular assist devices serving as mechanical circulatory support for patients with end-stage heart failure are associated with complications, including bleeding, thromboembolism, and infection. Axial-flow pumps might overcome some of these shortcomings. Here we report our experience with long-term application of the DeBakey VAD (MicroMed Technology, Inc, Houston, Tex). METHODS: Nine male transplant candidates (37 +/- 14 years) with severe hemodynamic compromise (cardiac index, 1.6 +/- 0.5 L . min(-1) x m(-2); pulmonary capillary wedge pressure, 27 +/- 6 mm Hg) and beginning end-organ failure despite inotropic and intra-aortic balloon pump support received the DeBakey VAD. Clinical outcome was evaluated. RESULTS: Cumulative support was 7.8 years, and the mean duration of support was 314 +/- 75 days (range, 229-438 days). Eight patients were transplanted, and one died from intracerebral bleeding. Peripheral circulation and end-organ function recovered rapidly after implantation. Continuous flow was able to maintain adequate organ perfusion over the long term. Eight patients were discharged during support, with good quality of life. There were no early bleedings, but there were late bleedings in 3 patients caused by excessive anticoagulation and platelet inhibition. Neurologic events occurred in 4 patients. Three patients recovered completely from symptoms, and one had lethal intracerebral bleeding. Because of thrombus formation, the device was exchanged in 4 patients. With increasing experience, thrombolysis was performed in similar situations. All such patients underwent successful transplantation. Hemolysis occurred, with events indicating thrombus formation. Device-related infection was found in one patient. CONCLUSIONS: The DeBakey VAD demonstrated its potential for long-term bridge to transplantation. The risk for thrombus formation needs to be addressed by improvement of pump technology and new strategies for platelet inhibition. PMID- 16214530 TI - Perivascular adipose tissue modulates vascular function in the human internal thoracic artery. AB - OBJECTIVE: Recent studies have shown that perivascular adipose tissue from the rat aorta secretes a substance that can dilate the aorta. The purpose of the present study was to examine whether this vasodilator is also present in human internal thoracic arteries. METHODS: Vascular function of human internal thoracic arteries with and without perivascular adipose tissue was assessed with wire myography, and morphology was examined with light microscopy. RESULTS: The presence of perivascular adipose tissue attenuated the maximal contraction to U 46619 and the contraction to phenylephrine (1 micromol/L) by 37% and 24%, respectively. Transfer of the solution incubated with a perivascular adipose tissue-intact vessel (donor) to a vessel without perivascular adipose tissue (recipient) induced a significant relaxation (36%) in the recipient artery precontracted with phenylephrine. Transfer of incubation solution with perivascular adipose tissue alone also induced a relaxation response in the recipient vessel (37%). The relaxation of the recipient artery induced by the transfer of incubation solution from the donor (artery with intact perivascular adipose tissue or perivascular adipose tissue alone) was absent in vessels precontracted by KCl (60 mmol/L) and was prevented by calcium-dependent potassium channel blockers (tetraethylammonium chloride, 1 mmol/L; iberiotoxin, 100 nmol/L), but not by the voltage-dependent potassium channel blocker 4 aminopyridine (1 mmol/L) and the adenosine triphosphate-dependent potassium channel blocker glibenclamide (10 micromol/L). CONCLUSIONS: Perivascular adipose tissue in human internal thoracic arteries releases a transferable relaxation factor that acts through the activation of calcium-dependent potassium channels. Because perivascular adipose tissue is often removed in coronary artery bypass grafting, retaining perivascular adipose tissue might be helpful in reducing the occurrence of vasospasm of the graft vessels. PMID- 16214531 TI - Reporting and classification of patient safety events in a cardiothoracic intensive care unit and cardiothoracic postoperative care unit. AB - OBJECTIVES: The objective was to evaluate a new mechanism for reporting and classifying patient safety events to increase reporting and identify patient safety priorities. METHODS: A voluntary patient safety event reporting system accessible by all health care workers was implemented in the Cardiothoracic Intensive Care and Post Anesthesia Care Units. Information collected included patient identifiers; date, time, and location of report and event; type and description of event; and severity score. Narrative descriptions of events were analyzed and coded to describe when in the care process the event occurred, what occurred, and a causal classification of why the event occurred. RESULTS: A total of 163 reports describing 157 events were received. These included 121 events reported from the intensive care unit (25.3 reported events per 1000 patient days), a 3-fold increase compared with the preexisting on-line reporting system. A total of 113 reports (69%) came from nurses, 31 from physicians (19%), and 10 from other staff (6%). A majority of events (85, 54%) reached the patient but caused no harm. Multiple causes were identified for the majority of events. The most frequent causes were related to human factors (48%) and organizational factors (34%). CONCLUSIONS: Health care workers were willing to use the patient safety event reporting system, which yielded a broad range of patient safety data. Patient safety events are multifaceted and often have multiple causal factors. Application of a causal classification model for patient safety event coding in the intensive care and preoperative and postoperative care units is feasible and facilitates local communication of important event-related information. PMID- 16214532 TI - Hyperglycemia during cardiopulmonary bypass is an independent risk factor for mortality in patients undergoing cardiac surgery. AB - BACKGROUND: Hyperglycemia is commonly present in the perioperative period in patients undergoing cardiac surgery, even during administration of insulin. A direct relationship between postoperative hyperglycemia and mortality has been established in diabetic patients undergoing cardiac surgery. However, this relationship might be confounded because patients with poor outcome receive more glucogenic drugs postoperatively. We assessed the influence of hyperglycemia (highest glucose level) during cardiopulmonary bypass on perioperative morbidity and mortality in diabetic and nondiabetic patients. METHODS: We performed a multivariate logistic regression analysis on all diabetic (n = 1579) and nondiabetic (n = 4701) patients undergoing cardiac surgery at the Toronto General Hospital between 1999 and 2001. Boluses of insulin were given during cardiopulmonary bypass when the glucose level exceeded 15 mmol/L, when the serum potassium level exceeded 6.0 mmol/L, or both. RESULTS: Overall mortality was 1.8% (n = 115). A high glucose level during cardiopulmonary bypass was an independent predictor of mortality in both diabetic (odds ratio, 1.20; confidence interval, 1.08-1.32) and nondiabetic (odds ratio, 1.12; confidence interval, 1.06-1.19; per millimole per liter increase in glucose) patients. A high glucose level during cardiopulmonary bypass was also an independent predictor of all major adverse events in both patient groups (odds ratio, 1.06; confidence interval, 1.03-1.09). A high glucose level was not closely related to cardiopulmonary bypass (r = 0.3) or aortic crossclamp times (r = 0.4). CONCLUSIONS: A high peak serum glucose level during cardiopulmonary bypass is an independent risk factor for death and morbidity in diabetic patients and unexpectedly also in nondiabetic patients. PMID- 16214533 TI - Cardioplegia prevents ischemia-induced transcriptional alterations of cytoprotective genes in rat hearts: a DNA microarray study. AB - BACKGROUND: Energy conservation and calcium homeostasis contribute to myocardial protection provided by hyperkalemic cardioplegia during ischemia. Complimenting these established mechanisms of protection, previous work suggested that activation of cytoprotective signaling pathways also contributes to reduced injury with cardioplegia. We proposed that cardioplegia would recruit cytoprotective pathways and investigated the transcriptional response of the heart after cardioplegia-protected ischemia compared with that after ischemia alone. METHODS: Isolated perfused rat hearts underwent 40 minutes of global ischemia alone or with St Thomas cardioplegia, followed by 120 minutes of reperfusion. The expression profiles of isolated RNA were determined by using Affymetrix microarrays and assessed by comparing cardioplegia-protected hearts and hearts undergoing unprotected ischemia with time-matched control hearts. The content of selected proteins was assessed by means of immunoblotting. RESULTS: Cardioplegia preserved the expression of multiple genes involved in carbohydrate and fatty acid metabolism, glycolysis, and electron transport compared with ischemia alone. The expression of the sodium-calcium exchanger and ryanodine receptor was preserved in line with the ability of cardioplegia to decrease calcium overload. The expression of multiple cytoprotective molecules, including protein-tyrosine kinase, calcineurin B, p38 mitogen-activated protein kinase, voltage-dependent anion channel, protein kinase C , heat shock protein 70, and manganese superoxide dismutase all showed decreased expression in ischemia but were preserved to near nonischemic levels by cardioplegia. CONCLUSION: Cardioplegia during ischemia maintained an expression profile similar to that seen in nonischemic hearts for genes involved in energy conservation, calcium homeostasis, and cytoprotective pathways, whereas ischemia alone did not. Exposing the transcriptional differences in cytoprotective genes during untreated and cardioplegia-treated ischemia provides valuable insight into an additional mechanism of cardioprotection induced by cardioplegia. PMID- 16214534 TI - High level of cerebral microembolization in patients supported with the DeBakey left ventricular assist device. AB - OBJECTIVE: Microembolic signals detected by transcranial Doppler ultrasonography have been demonstrated to be clinically relevant in patients supported with pulsatile left ventricular assist devices. We prospectively investigated the quantity of microembolic signals in patients supported with the continuous-flow DeBakey left ventricular assist device (MicroMed DeBakey VAD; MicroMed Technology, Inc, Houston, Tex) including the refined Carmeda BioActive Surface system (Carmeda AB, Stockholm, Sweden). METHODS: Twenty-three patients (20 male) aged 14 to 62 years supported with DeBakey left ventricular assist devices (n = 6 with Carmeda) were enrolled in this study. Microembolic signal monitorings were performed twice weekly by insonating the middle cerebral artery for 20 minutes without and 20 minutes with oronasal application of oxygen (6 L/min). Evidence of clinically manifest thromboembolic events was based on regular questionnaires, clinical examinations, and results of diagnostic procedures. RESULTS: Despite a low incidence of thromboembolic complications (0.24 per 100 left ventricular assist device days), 20 patients (87%) showed circulating microemboli. Overall, microembolic signals were found in 175 of 499 transcranial Doppler ultrasonographic examinations (35.1%), with mean counts of 81.2 +/- 443 (range 0 5042 signals/h). Both microembolic signal prevalence (25% vs 34%, P = .01) and absolute signal counts (46.5 vs 104, P < .01) significantly declined with oxygen delivery. There was no significant correlation between the individual microembolic signal activity and the incidence of clinical thromboembolism or the intensity of antihemostatic treatment. Patients supported with the Carmeda device did not show reduced rates of clinical thromboembolization or cerebral microemboli. CONCLUSION: In patients with DeBakey left ventricular assist devices, a high load of clinically silent microemboli can be detected within the cerebral arteries despite a low incidence of embolic complications. It needs to be investigated whether such continuous, presumably gaseous microembolization causes cognitive or neuropsychologic deficits. PMID- 16214535 TI - Selective cyclooxygenase 2 inhibitor induces indefinite survival of fully allogeneic cardiac grafts and generates CD4+ regulatory cells. AB - BACKGROUND: Selective inhibition of cyclooxygenase 2 has been reported to have not only anti-inflammatory effects but also effects on the immune response. We investigated ability of a cyclooxygenase 2 inhibitor to inhibit alloimmune response in a murine cardiac transplantation model. METHODS: CBA (H2(k)) mice underwent transplantation of C57BL/10 (H2(b)) hearts. On the day of transplantation, the recipients received either no treatment or single administration of aspirin (a cyclooxygenase 1 and 2 inhibitor) or the selective cyclooxygenase 2 inhibitor NS-398. Naive CBA mice (secondary recipients) underwent adoptive transfer of splenocytes from treated mice with long-surviving grafts (primary recipients) to determine whether regulatory cells developed after NS-398 treatment. Histologic, cell-proliferation, and cytokine studies were also performed. RESULTS: Untreated CBA mice rejected C57BL/10 cardiac grafts acutely (median survival time, 8 days). The majority of recipients given aspirin rejected their grafts within 20 days (median survival time, 11 days). In mice given NS 398, the majority of the grafts survived indefinitely (median survival time, >100 days). Secondary CBA recipients given CD4+ splenocytes from primary CBA recipients treated with NS-398 also had indefinite survival of C57BL/10 hearts (median survival time, >60 days). Graft acceptance and proliferative hyporesponsiveness were also confirmed by the histologic and cell-proliferation studies, respectively. Production of interleukin 4 and 10 from splenocytes of the recipients treated with NS-398 were significantly higher than that from untreated recipients. CONCLUSIONS: In our model administration of cyclooxygenase 2 inhibitor induced indefinite survival of fully mismatched cardiac grafts and generated CD4+ regulatory cells. Cyclooxygenase 2 inhibitor could warrant consideration for use as an immunomodulating agent in clinical transplantation. PMID- 16214536 TI - Persistently increased systemic, but not cardiac-specific, adhesion molecule expression and coronary endothelial dysfunction in human cardiac allografts. AB - BACKGROUND: Adhesion molecules are involved in inflammatory processes that alter endothelial function and lead to impairment of coronary vasomotor function. We studied a possible relationship between systemic expression, cardiac-specific expression, or both of P-selectin and intercellular adhesion molecule 1 and coronary vasomotor function both 1 and 12 months after heart transplantation in human subjects. METHODS: The expression of endomyocardial and soluble forms of P selectin and intercellular adhesion molecule 1, as well as levels of tumor necrosis factor alpha, were determined in aortic and coronary sinus blood samples 1, 6, and 12 months after heart transplantation in 42 transplant recipients and 20 age-matched, nontransplanted control subjects. In addition, both endothelium dependent (acetylcholine) and endothelium-independent (adenosine) coronary vasomotor function were assessed by using a Doppler flow wire and quantitative coronary angiography 1 and 12 months after heart transplantation. RESULTS: Adhesion molecules were highly expressed 1 month after heart transplantation and remained at high levels 12 months after heart transplantation when compared with levels in nontransplanted control subjects. No cardiac-specific expression or release of P-selectin or intercellular adhesion molecule 1 was observed. There was a significant inverse correlation between coronary vasomotor function and soluble adhesion molecule expression both 1 and 12 months after heart transplantation. CONCLUSION: Persistently high levels of circulating adhesion molecules are of systemic, but not cardiac-specific, origin and reflect a chronic inflammatory state throughout the first year after heart transplantation. This is associated with impairment of coronary vasomotor function, an early and potentially reversible step in the process of atherothrombosis and transplant coronary artery disease. PMID- 16214537 TI - Role of the beta-adrenergic receptor kinase in myocardial dysfunction after brain death. AB - OBJECTIVE: Significant cardiac dysfunction after brain death leading to exclusion from procurement for cardiac transplantation is seen in up to 25% of potential organ donors in the absence of structural heart disease. The cause includes uncoupling of the myocardial beta-adrenergic receptor signaling system. The mechanism, however, has not yet been described. This study investigates our hypothesis that brain death causes acute activation of the betaAR kinase and leads to desensitization of myocardial beta-adrenergic receptors and impaired ventricular function. METHODS: Adult pigs underwent a sham operation or induction of brain death by means of subdural balloon inflation (n = 8 in each group). Cardiac function was assessed by using sonomicrometry at baseline and for 6 hours after the operation. beta-Adrenergic receptor signaling was assessed at 6 hours after the operation by measuring myocardial sarcolemmal membrane adenylate cyclase activity, beta-adrenergic receptor density, beta-adrenergic receptor kinase expression, and activity. RESULTS: Induction of brain death led to significantly decreased left ventricular systolic and diastolic function. Basal and isoproterenol-stimulated adenylate cyclase activity was blunted in the brain dead group compared with the sham-operated group (28.3 +/- 4.3 vs 48.3 +/- 7.6 pmol of cyclic adenosine monophosphate.mg(-1) x min(-1) [P = .01] and 54.8 +/- 9.6 vs 114.5 +/- 18 pmol of cyclic adenosine monophosphate x mg(-1) x min(-1) [P < .02]). There was no difference in beta-adrenergic receptor density between the brain dead and sham-operated groups. Myocardial beta-adrenergic receptor kinase expression was 3-fold greater in the brain dead versus sham-operated groups, and membrane beta-adrenergic receptor kinase activity was 2.5-fold greater in the brain dead group compared with that seen in the sham-operated group. CONCLUSION: Induction of brain death leads to significant left ventricular dysfunction in this porcine model. Cardiac beta-adrenergic receptors are clearly uncoupled after brain death, and our data suggest that the mechanism is acute increase of myocardial beta-adrenergic receptor kinase activity, leading to beta-adrenergic receptor desensitization and ventricular dysfunction. PMID- 16214538 TI - Clinical-pathologic conference in general thoracic surgery: malignant transformation of recurrent respiratory papillomatosis. PMID- 16214539 TI - Diagnosis of ruptured pulmonary hydatid cyst by means of flexible fiberoptic bronchoscopy: a report of three cases. PMID- 16214540 TI - Clear cell carcinoma of the lung revisited. PMID- 16214541 TI - Successful enucleation of a giant cardiac hemangioendothelioma showing an unusual proliferation pattern. PMID- 16214542 TI - Lymphangiomatous hamartoma: cause or bystander of the isolated chylopericardium? PMID- 16214543 TI - Transcatheter closure of patent ductus arteriosus with the Inoue single-branched stent graft. PMID- 16214544 TI - Induction chemotherapy, surgical resection, and high-dose chemotherapy for mediastinal nonseminomatous germ-cell tumor. PMID- 16214545 TI - One-stage definitive repair of pulmonary atresia with intact ventricular septum and hypoplastic right ventricle. PMID- 16214546 TI - The "clover technique" as a new approach for correction of postendocarditic severe tricuspid valve regurgitation. PMID- 16214547 TI - Left main coronary artery atresia masquerading as dilated cardiomyopathy treated with aortic reimplantation. PMID- 16214548 TI - Separation of thoracopagus-cardiopagus twins joined by a myocardial bridge. PMID- 16214549 TI - Thoracoscopic removal of a knife impaled in the chest. PMID- 16214550 TI - Aortic valve-sparing operation for autograft failure after the Ross procedure. PMID- 16214551 TI - A giant pleural fibrous tumor. PMID- 16214552 TI - Left-sided modified Kergin carinoplasty as an alternative to carinal resection for centrally located non-small cell lung cancer pretreated by radiochemotherapy. PMID- 16214553 TI - Thoracoscopic operation with local and epidural anesthesia in the treatment of pneumothorax after lung transplantation. PMID- 16214555 TI - Emergency surgical intervention in a patient with delayed diagnosis of aortic dissection presenting with acute ischemic stroke and undergoing thrombolytic therapy. PMID- 16214554 TI - Chronic dissection of the right aortic sinus with detachment of the aortic annulus and coronary artery. PMID- 16214556 TI - Combined surgical and endoprothesical approach to solve a thoracic aortic pseudoaneurysm 15 years after coarctation correction. PMID- 16214558 TI - Evaluation of surgical trauma and cardiopulmonary bypass as factors in inflammatory status after cardiac surgery. PMID- 16214559 TI - Radial artery for coronary artery bypass grafting. PMID- 16214562 TI - Anatomy of the orbit and its related structures. PMID- 16214563 TI - Patient evaluation. PMID- 16214564 TI - The diagnosis and management of dry eyes. PMID- 16214566 TI - Current techniques of entropion and ectropion correction. PMID- 16214565 TI - Botulinum toxin management of upper facial rhytidosis and blepharospasm. PMID- 16214567 TI - Ptosis evaluation and management. PMID- 16214568 TI - Comprehensive management of eyebrow and forehead ptosis. PMID- 16214569 TI - Reconstructive blepharoplasty. PMID- 16214570 TI - The diagnosis and management of blepharoplasty complications. PMID- 16214571 TI - Reconstruction of the upper eyelid. PMID- 16214572 TI - Lower eyelid reconstruction. PMID- 16214573 TI - Diagnosis and management of thyroid orbitopathy. PMID- 16214574 TI - Surgical management of essential blepharospasm. PMID- 16214575 TI - Endoscopic lacrimal surgery. PMID- 16214576 TI - Facial lifting with "APTOS" threads: featherlift. PMID- 16214577 TI - Cosmetic uses of injectable phosphatidylcholine on the face. PMID- 16214578 TI - The NHS: a national health sham. PMID- 16214580 TI - Turkey behind closed doors. PMID- 16214579 TI - Progress in TB vaccine development. PMID- 16214581 TI - The PROactive study: some answers, many questions. PMID- 16214582 TI - Suicide in prison. PMID- 16214583 TI - Internet-based eye care: VISION 2020. PMID- 16214584 TI - Clean Care is Safer Care: a worldwide priority. PMID- 16214585 TI - Refining the exercise prescription for hypertension. PMID- 16214586 TI - International health regulations (2005). PMID- 16214587 TI - Naming seizures. PMID- 16214588 TI - Judah Folkman: persistent pioneer in cancer research. PMID- 16214589 TI - Misleading data analyses in salmeterol (SMART) study. PMID- 16214591 TI - A stain on medical ethics. PMID- 16214592 TI - A stain on medical ethics. PMID- 16214593 TI - Prostitutes are people too. PMID- 16214594 TI - Missed contraceptive pill recommendations. PMID- 16214595 TI - Screening for oral cancer. PMID- 16214597 TI - Efficacy and safety of cholesterol-lowering treatment: prospective meta-analysis of data from 90,056 participants in 14 randomised trials of statins. AB - BACKGROUND: Results of previous randomised trials have shown that interventions that lower LDL cholesterol concentrations can significantly reduce the incidence of coronary heart disease (CHD) and other major vascular events in a wide range of individuals. But each separate trial has limited power to assess particular outcomes or particular categories of participant. METHODS: A prospective meta analysis of data from 90,056 individuals in 14 randomised trials of statins was done. Weighted estimates were obtained of effects on different clinical outcomes per 1.0 mmol/L reduction in LDL cholesterol. FINDINGS: During a mean of 5 years, there were 8186 deaths, 14,348 individuals had major vascular events, and 5103 developed cancer. Mean LDL cholesterol differences at 1 year ranged from 0.35 mmol/L to 1.77 mmol/L (mean 1.09) in these trials. There was a 12% proportional reduction in all-cause mortality per mmol/L reduction in LDL cholesterol (rate ratio [RR] 0.88, 95% CI 0.84-0.91; p<0.0001). This reflected a 19% reduction in coronary mortality (0.81, 0.76-0.85; p<0.0001), and non-significant reductions in non-coronary vascular mortality (0.93, 0.83-1.03; p=0.2) and non-vascular mortality (0.95, 0.90-1.01; p=0.1). There were corresponding reductions in myocardial infarction or coronary death (0.77, 0.74-0.80; p<0.0001), in the need for coronary revascularisation (0.76, 0.73-0.80; p<0.0001), in fatal or non-fatal stroke (0.83, 0.78-0.88; p<0.0001), and, combining these, of 21% in any such major vascular event (0.79, 0.77-0.81; p<0.0001). The proportional reduction in major vascular events differed significantly (p<0.0001) according to the absolute reduction in LDL cholesterol achieved, but not otherwise. These benefits were significant within the first year, but were greater in subsequent years. Taking all years together, the overall reduction of about one fifth per mmol/L LDL cholesterol reduction translated into 48 (95% CI 39-57) fewer participants having major vascular events per 1000 among those with pre-existing CHD at baseline, compared with 25 (19-31) per 1000 among participants with no such history. There was no evidence that statins increased the incidence of cancer overall (1.00, 0.95-1.06; p=0.9) or at any particular site. INTERPRETATION: Statin therapy can safely reduce the 5-year incidence of major coronary events, coronary revascularisation, and stroke by about one fifth per mmol/L reduction in LDL cholesterol, largely irrespective of the initial lipid profile or other presenting characteristics. The absolute benefit relates chiefly to an individual's absolute risk of such events and to the absolute reduction in LDL cholesterol achieved. These findings reinforce the need to consider prolonged statin treatment with substantial LDL cholesterol reductions in all patients at high risk of any type of major vascular event. PMID- 16214599 TI - Effect of BCG revaccination on incidence of tuberculosis in school-aged children in Brazil: the BCG-REVAC cluster-randomised trial. AB - BACKGROUND: Many countries offer a second BCG vaccination to prevent tuberculosis, although there is little evidence of whether this confers additional protection. BCG vaccination is routine in Brazil but BCG revaccination procedures vary by state. We studied revaccination efficacy in two Brazilian cities with tuberculosis prevalence representative of Brazil. METHODS: We did a cluster-randomised trial of the protection against tuberculosis from BCG revaccination in school-aged children who had had one BCG vaccination as infants. 767 schools in the cities of Salvador and Manaus, Brazil, participated; schools were the unit of randomisation. The study was open label with no placebo. Cases of tuberculosis were identified through record linkage to the Tuberculosis Control Programme. Revaccination status was masked during linkage and validation of cases. The incidence of tuberculosis was the primary outcome. Analysis was by intention to treat. FINDINGS: 386 schools (176,846 children) were assigned BCG revaccination and 365 (171,293 children) no revaccination. 42,053 children in the vaccine group and 47,006 in the control group were absent from school on the day of the visit and were excluded. 31,163 and 27,146, respectively were also excluded because they had no BCG scar, two or more scars, or a doubtful scar on assessment. The crude incidence of tuberculosis in the intervention group was 29.3 per 100,000 person years and in the control group 30.2 per 100,000 person years (crude-rate ratio 0.97; 95% CI 0.76-1.28). The efficacy of BCG revaccination was 9% (-16 to 29%). INTERPRETATION: Revaccination given to children aged 7-14 years in this setting does not provide substantial additional protection and should not be recommended. Follow-up is ongoing and needed to assess the effect of other factors on revaccination efficacy: time since vaccination, age at vaccination, and high or low prevalence of environmental mycobacteria. PMID- 16214600 TI - Infection with Chlamydia trachomatis after mass treatment of a trachoma hyperendemic community in Tanzania: a longitudinal study. AB - BACKGROUND: Data from studies done in communities where trachoma is mesoendemic suggest that ocular infection with Chlamydia trachomatis can be eliminated after one mass treatment with antibiotics. However, there are no comparable long-term data from trachoma hyperendemic communities. Our aim, therefore, was two-fold: first, to ascertain the disease pattern of trachoma and ocular infection with C trachomatis in a trachoma hyperendemic community after mass treatment; and, second, to ascertain the risk factors for incident infection. METHODS: We did a longitudinal study of a trachoma hyperendemic community (n=1017) in Tanzania. We did surveys, including ocular swabs, at baseline, 2, 6, 12, and 18 months to identify the presence, and quantity, of C trachomatis after single mass treatment of all individuals aged 6 months or older with azithromycin 20 mg per kg; pregnant women without clinical disease received topical tetracycline. FINDINGS: Mass treatment (coverage 86%) significantly reduced the prevalence of infection from 57% (495 of 871) to 12% (85 of 705) at 2 months. Infection remained fairly constant to 12 months, with evidence of increasing numbers and load of infection by 18 months post-treatment. Incident infection at 6 months was 3.5-times more likely if another member of the household had more than 19 organisms per swab at 2 months. Travel outside the village, and visitors to the household, did not increase the risk of infection within households up to 12 months. INTERPRETATION: In this trachoma hyperendemic community, infection levels after high antibiotic coverage persisted at a low level to 18 months, with evidence for re-emergence after 1 year. Fairly light loads of infection were associated with household transmission. Yearly mass treatment over a few years could be sufficient to eliminate infection. PMID- 16214601 TI - Suicides in male prisoners in England and Wales, 1978-2003. AB - The number of suicides in English and Welsh prisons is increasing, but the excess compared with the general population has not been reliably quantified. We therefore compared, in narrow age bands, all 1312 suicides of male prisoners in England and Wales between 1978 and 2003 with suicide rates in the general male population. The overall standardised mortality ratio for suicide was 5.1 (95% CI, 4.8-5.3), suggesting a five-fold excess of suicides in male prisoners, with a particularly striking excess in boys aged 15-17 years (standardised mortality ratio 18 [13-26]). The proportional excess of suicides of male prisoners has been increasing during the past quarter of a century, which underscores the need for substantial improvements in suicide prevention in prisons. PMID- 16214602 TI - Cholangiocarcinoma. AB - Cholangiocarcinoma is a devastating malignancy that presents late, is notoriously difficult to diagnose, and is associated with a high mortality. The incidence of intrahepatic cholangiocarcinoma is increasing worldwide. The cause for this rise is unclear, although it could be related to an interplay between predisposing genetic factors and environmental triggers. MRI and CT with endoscopic ultrasound and PET provide useful diagnostic information in certain patients. Surgical resection is the only chance for cure, with results depending on careful technique and patient selection. Data suggest that liver transplantation could offer long-term survival in selected patients when combined with neoadjuvant chemoradiotherapy. Chemotherapy and radiotherapy have been ineffective for patients with inoperable tumours. For most of these patients biliary drainage is the mainstay of palliation. However, controversy exists over the type and positioning of biliary stents. Photodynamic treatment is a new palliative technique that might improve quality of life. PMID- 16214603 TI - What makes a good genetic association study? AB - Genetic association studies are central to efforts to identify and characterise genomic variants underlying susceptibility to multifactorial disease. However, obtaining robust replication of initial association findings has proved difficult. Much of this inconsistency can be attributed to inadequacies in study design, implementation, and interpretation--inadequately powered sample groups are a major concern. Several additional factors affect the quality of any given association study, with appropriate sample-recruitment strategy, logical variant selection, minimum genotyping error, relevant data analysis, and valid interpretation all essential to generation of robust findings. Replication has a vital role in showing that associations that are identified reflect interesting biological processes rather than methodological quirks. For an unbiased view of the evidence for and against any particular association, study quality, rather than significance value, needs to play the dominant part. PMID- 16214604 TI - A public-health campaign to raise awareness of children's wellbeing with images drawn by children. PMID- 16214605 TI - Adolescent siblings with thigh pain at the same age. PMID- 16214606 TI - A life in the day of a voluntary service overseas lecturer in Guyana. PMID- 16214607 TI - Fast frequency selective MR imaging. AB - With the proposed fast frequency selective MR imaging (FFSMRI) method, we focused on the elimination of all off-resonance components from the image of the observed object. To maintain imaging speed and simultaneously achieve good frequency selectivity, MRI was divided into two steps: signal acquisition and postprocessing. After the preliminary phase in which we determine imaging parameters, MRI takes place; the signal from the same object is successively acquired M times. As a result, we obtain M partial signals in k-space, from which we calculate the image of the observed object in postprocessing phase, after signal acquisition has been completed. With proper selection of parameters, it is possible to exclude from the image a majority of off-resonance components present in the observed object. However, we can decide to keep only a chosen off resonance component in the image and eliminate all other components, including the on-resonance component and thus producing a different image from the same acquisition. The experiments with Fe(OH)(3) and oil showed that signal-to-noise ratio (SNR) can be improved by about a factor of four. The proposed FFSMRI method is suitable for frequency selective MR imaging and quantitative measurements in dynamic MRI where exclusion of off-resonance components can improve the reliability of measurement. PMID- 16214608 TI - Matrix-based autologous chondrocyte implantation for cartilage repair: noninvasive monitoring by high-resolution magnetic resonance imaging. AB - OBJECTIVE: Monitoring of articular cartilage repair after matrix-associated autologous chondrocyte implantation (MACI) by a new grading and point-scale system based on noninvasive cartilage-specific magnetic resonance imaging (MRI) protocol. PATIENTS AND METHODS: In 20 patients, postoperative high-resolution MRI follow-up examinations at 4, 12, 24 and 52 weeks after matrix-based ACI for cartilage repair were initiated. The repair tissue was described with separate variables: degree of defect repair in width and length, surface, structure and signal intensity of the repair tissue, and status of the subchondral lamina and bone. For these variables, a grading system with point-scale evaluation was applied, and the mean average values were calculated for every follow-up MR exam of each patient. RESULTS: In 10 patients, an incomplete filling of the defect improved to complete filling (6 patients) or less incomplete (4 patients) filling of the defect. Three cases of implant hypertrophy returned to normal within 1 year. Complete filling of the defect by repair tissue was found in 2 patients from the beginning. Integration was complete in 10 cases. Improvement of incomplete to complete integration was found in 3 patients. The signal intensity of the implant developed to native cartilage signal in 13 patients. The mean average values increased from the 4th to the 52 nd week in 17 of 20 patients and decreased in 3 of 20 patients. CONCLUSION: High-resolution MRI provides a noninvasive tool for monitoring the development of cartilage repair tissue in MACI over time and helps to differentiate abnormal repair tissue from a normal maturation process. PMID- 16214609 TI - MR safety and compatibility of a noninvasively expandable total-joint endoprosthesis. AB - A noninvasively expandable total-joint endoprosthesis is now available for pediatric patients; the prosthesis can be lengthened by external application of a magnetic field. We investigated the risks of unintentional heating or lengthening of the prosthesis during MR imaging and evaluated the effect of the device on the diagnostic efficacy of MR imaging of surrounding tissues. We performed MR imaging at 1.5 T by using standard pulse sequences and pulse sequences with high-gradient and high-radiofrequency duty cycle. MR imaging caused no measurable change in prosthesis length, and the temperature of the prosthesis increased by less than 1 degrees C during repeated 14-min exposures. Despite significant signal loss and image distortion around the prosthetic joint, clinically useful images were obtained as close as 12 cm from the ends of the prosthetic stems, measured toward the body of the device. Thus, the prosthesis can be safely exposed to MR imaging pulse sequences at 1.5 T, and the visualization of some tissue surrounding the device is clinically useful. PMID- 16214610 TI - Comparison of different methods for combining phase-contrast images obtained with multiple coils. AB - The ability to determine coil sensitivities implies that a method optimized in terms of maximized signal-to-noise ratio (SNR) can be applied to the combination of multiple coil images. An optimization of SNR subsequently results in a minimized variance in quantitative velocity measurements using phase-contrast imaging. When coil sensitivities are unknown, the weighted mean method, utilizing the square of the signal magnitude as weights, is suitable for combination of multiple phase images. In this study, the optimized method using estimated coil sensitivities was compared to the weighted mean method both theoretically and experimentally. It is shown that absence of noise correlation between the different coil images implies no difference between the methods regarding the variance of the phase. In the practical situation, noise correlation does exist, implying an opportunity for further reduction of phase variance using the optimized method. In vitro and in vivo studies showed, however, no significant difference between the two methods studied. PMID- 16214611 TI - An improved 3-D Look--Locker imaging method for T(1) parameter estimation. AB - The 3-D Look-Locker (LL) imaging method has been shown to be a highly efficient and accurate method for the volumetric mapping of the spin lattice relaxation time T(1). However, conventional 3-D LL imaging schemes are typically limited to small tip angle RF pulses (5 degrees ), thereby improving the SNR and the accuracy of the method. In phantom studies, a mean T(1) measurement accuracy of less than 2% (0.2-3.1%) using a tip angle of 10 degrees was obtained for a range of T(1) from approximately 300 to 1,700 ms with a measurement time increase of only 15%. This accuracy compares favorably with the conventional 3-D LL method that provided an accuracy between 2.2% and 7.3% using a 5 degrees flip angle. PMID- 16214598 TI - Secondary prevention of macrovascular events in patients with type 2 diabetes in the PROactive Study (PROspective pioglitAzone Clinical Trial In macroVascular Events): a randomised controlled trial. AB - BACKGROUND: Patients with type 2 diabetes are at high risk of fatal and non-fatal myocardial infarction and stroke. There is indirect evidence that agonists of peroxisome proliferator-activated receptor gamma (PPAR gamma) could reduce macrovascular complications. Our aim, therefore, was to ascertain whether pioglitazone reduces macrovascular morbidity and mortality in high-risk patients with type 2 diabetes. METHODS: We did a prospective, randomised controlled trial in 5238 patients with type 2 diabetes who had evidence of macrovascular disease. We recruited patients from primary-care practices and hospitals. We assigned patients to oral pioglitazone titrated from 15 mg to 45 mg (n=2605) or matching placebo (n=2633), to be taken in addition to their glucose-lowering drugs and other medications. Our primary endpoint was the composite of all-cause mortality, non fatal myocardial infarction (including silent myocardial infarction), stroke, acute coronary syndrome, endovascular or surgical intervention in the coronary or leg arteries, and amputation above the ankle. Analysis was by intention to treat. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN NCT00174993. FINDINGS: Two patients were lost to follow-up, but were included in analyses. The average time of observation was 34.5 months. 514 of 2605 patients in the pioglitazone group and 572 of 2633 patients in the placebo group had at least one event in the primary composite endpoint (HR 0.90, 95% CI 0.80-1.02, p=0.095). The main secondary endpoint was the composite of all cause mortality, non-fatal myocardial infarction, and stroke. 301 patients in the pioglitazone group and 358 in the placebo group reached this endpoint (0.84, 0.72 0.98, p=0.027). Overall safety and tolerability was good with no change in the safety profile of pioglitazone identified. 6% (149 of 2065) and 4% (108 of 2633) of those in the pioglitazone and placebo groups, respectively, were admitted to hospital with heart failure; mortality rates from heart failure did not differ between groups. INTERPRETATION: Pioglitazone reduces the composite of all-cause mortality, non-fatal myocardial infarction, and stroke in patients with type 2 diabetes who have a high risk of macrovascular events. PMID- 16214612 TI - Representative brain selection using a group-specific tissue probability map. AB - PURPOSE: To determine the anatomy of a standard brain reflecting well-defined group characteristics based on probabilistic information from group-specific anatomical variations. MATERIALS AND METHODS: We constructed a group-specific tissue probabilistic map for 20 subjects and used it to extract voxel-wise probabilistic information for each subject through regional spatial normalization using Automated Image Registration software (AIR 5.2.5). Extracted probabilistic information was then used to determine standard properties of the subjects. For comparison, we employed an empirical scoring function - a measure of entropy - in ordering the data set. A brain with minimum entropy was then selected for a group standard. The evaluation of our proposed method was performed using two different selection schemes: deformation analysis and similarity index measurements. RESULTS: This method showed highly correlated result with previous method by Kochunov et al., with fewer computational tasks. CONCLUSION: This method can thus be used to determine an appropriate standard model to compare with disease affected brains. PMID- 16214613 TI - A soft-segmentation visualization scheme for magnetic resonance images. AB - Prevalent visualization tools exploit gray value distribution in images through modified histogram equalization and matching technique, referred to as the window width/window level-based method, to improve visibility and enhance diagnostic value. The window width/window level tool is extensively used in magnetic resonance (MR) images to highlight tissue boundaries during image interpretation. However, the identification of different regions and distinct boundaries between them based on gray-level distribution and displayed intensity levels is extremely difficult because of the large dynamic range of tissue intensities inherent in MR images. We propose a soft-segmentation visualization scheme to generate pixel partitions from the histogram of MR image data using a connectionist approach and then generate selective visual depictions of pixel partitions using pseudo color based on an appropriate fuzzy membership function. By applying the display scheme in clinical examples in this study, we could demonstrate additional overlapping regions between distinct tissue types in healthy and diseased areas (in the brain) that could help improve the tissue characterization ability of MR images. PMID- 16214614 TI - Manganese-enhanced MRI of rat spinal cord injury. AB - The potential of the manganese-enhanced MRI (MEI) technique in labeling the intact neuronal circuitry of rat spinal cord was examined. Experiments were conducted on normal and injured cords at 9.4-T magnetic field strength using an implantable rf coil. The contrast agent manganese (Mn) was locally delivered within the parenchyma at a dose of 25 mmol/L in 10 nL. The transport, uptake and accumulation of Mn in tissue were then followed remotely on T1-weighted images that were acquired serially from the cord. In MEIs of normal cord, Mn was observed to be transported in directions both rostral and caudal to the site of injection. In the cord that was subjected to hemisection, signal enhancement was on the contralesional side of the cord, but not at the ipsilesional side. The sensitivity and specificity of the MEI technique in labeling the neurons that are functional were also validated with a traditional track-tracing method using biotinylated dextran amine. PMID- 16214616 TI - Putting the evidence first. PMID- 16214617 TI - A lowering of the bar. PMID- 16214618 TI - Gateway offer overpriced. PMID- 16214619 TI - Postgraduate contract education: what defines quality of care? PMID- 16214620 TI - Memories. PMID- 16214621 TI - Three-dimensional localization of maxillary canines with cone-beam computed tomography. AB - INTRODUCTION: Precise 3-dimensional (3D) localization of impacted canines is central to their clinical management. Recently introduced dental 3D volumetric imaging systems make precise localization possible. The purpose of this study was to describe the spatial relationship of impacted canines by using images obtained with the NewTom QR-DVT 9000 (QR Srl, Verona, Italy). METHODS: Unilaterally and bilaterally impacted canines (n = 27) from 19 consecutive patients (15 female, 4 male) were evaluated on images taken with the NewTom QR-DVT 9000. The spatial relationships of the impacted canines relative to adjacent structures and incisor resorption were assessed with 3D visualization software. RESULTS: Most (92.6%) of the 27 impactions were palatal. Incisor resorption adjacent to the impacted canine was present in 66.7% of the lateral incisors and 11.1% of the central incisors. Follicle size did not play a major role in influencing impacted canine position. The alveolus was narrower at the impacted canine side compared with the erupted canine side; however, the width of the alveolus on the impacted canine side is independent of the deciduous canines. A correlation was found between the proximity of the impacted canine to the incisors and their resorption. There was no common location where eruption was arrested, and great variation in the inclination of the impacted canine was found. CONCLUSIONS: 3D volumetric imaging of impacted canines can show the following: presence or absence of the canine, size of the follicle, inclination of the long axis of the tooth, relative buccal and palatal positions, amount of the bone covering the tooth, 3D proximity and resorption of roots of adjacent teeth, condition of adjacent teeth, local anatomic considerations, and overall stage of dental development. In short, 3D imaging is clearly advantageous in the management of impacted canines. PMID- 16214622 TI - Reliability of measuring facial morphology with a 3-dimensional laser scanning system. AB - INTRODUCTION: The purpose of this prospective clinical trial was to evaluate the reliability of a 3-dimensional facial scanning technique for the measurement of facial morphology. METHODS: A field study was conducted in 2 comprehensive schools in the South Wales region of the United Kingdom. Forty subjects, mean age 11 years 3 months, were analyzed for soft tissue changes at baseline (T1), within 3 minutes (T2), and 3 days later (T3) by using 2 commercially available Minolta Vivid 900 (Osaka, Japan) laser-scanning devices assembled as a stereo pair. Left and right images were merged to form the whole face, and these images were superimposed to assess the errors at T1 and T2, and T1 and T3. RESULTS: The results showed that premerged left and right mean shell deviations were 0.38 +/- 0.14 mm for scans at T1, 0.31 +/- 0.09 mm at T2, and 0.34 +/- 0.12 mm at T3. The mean differences of the merged composite face were 0.31 +/- 0.08 mm between T1 and T2, and 0.40 +/- 0.11 mm between T1 and T3. Paired t tests showed no significant difference between these groups (P > .05). Shell deviation facial maps of the merged scans showed that 90% of the created composite facial scans were within an error of 0.85 mm. CONCLUSIONS: Capturing the soft tissue morphology of the face with this technique is clinically reproducible within 3 minutes and 3 days of the initial records. PMID- 16214623 TI - Comparison of peer assessment rating (PAR) index scores of plaster and computer based digital models. AB - INTRODUCTION: The peer assessment rating (PAR) index is a valid and reliable tool for measuring malocclusion on plaster models, but it has not been shown to be valid and reliable when used to score computer-based digital models. The purpose of this study was to determine whether the PAR index is a valid and reliable measure on digital models. METHODS: The study sample consisted of 48 pairs of plaster and digital pretreatment models. One examiner, calibrated in the PAR index, scored the digital and plaster models. The overall PAR scores were examined for reliability and validity by using analysis of variance and the intraclass correlation coefficient (ICC). Reliability of the components of the PAR score was compared with values originally presented by Richmond et al (1992). RESULTS: No significant differences were found between overall PAR scores of plaster and digital models (P = .82), and scores were highly correlated (ICC = 0.95; lower confidence boundary (LCB) = 0.92; upper confidence boundary (UCB) = 0.97). Intraexaminer reliability was excellent for both plaster models (ICC = 0.98; LCB = 0.97; UCB = 0.99) and digital models (ICC = 0.96; LCB = 0.94; UCB = 0.98). Reliability of all components of the PAR score generated on digital models except for buccal occlusion was similar to those of Richmond et al. CONCLUSION: PAR scores derived from digital models are valid and reliable measures of occlusion. PMID- 16214624 TI - The influence of a structured telephone call on orthodontic pain and anxiety. AB - INTRODUCTION: This purpose of this study was to examine the effects of a structured telephone call after orthodontic appliance placement on self-reported pain and anxiety. METHODS: One hundred-fifty orthodontic patients were randomly assigned to 1 of 3 groups and matched for age, sex, and ethnicity. The subjects completed baseline questionnaires to assess their levels of pain (on a 100-mm visual analog scale) and anxiety (Spielberger's State-Trait Anxiety Inventory) before orthodontic treatment. After the initial archwires were placed, all subjects completed the pain questionnaire and state-anxiety inventory at the same time daily for 1 week. One group also received a structured telephone call demonstrating care and reassurance; the second group received an attention-only telephone call, thanking them for participating in the study; the third group served as a control. RESULTS: Although both telephone groups reported significantly less pain (P = .005) and state-anxiety (P = .033) than the control group, there was no difference between the 2 telephone groups (P > .12 for pain; P > .81 for state-anxiety). CONCLUSIONS: A telephone call from a health-care provider reduced patients' self-reported pain and anxiety; the content of the telephone call was not important. PMID- 16214626 TI - Long-term profile changes in extraction and nonextraction patients. AB - INTRODUCTION: The purpose of this investigation was to determine the long-term differences in soft tissue profile changes between extraction and nonextraction patients who had been treated to the same incisor position and lip line. METHODS: Twenty extraction and 20 matched nonextraction patients, with posttreatment and long-term follow-up (average 15 years) records, were selected from a single private orthodontic practice. Posttreatment and long-term follow-up profile photos of the patients' nose, lip, and chin areas were evaluated by 105 orthodontists and 225 laypeople, who indicated their preferences and the amount of change they perceived among the 40 profiles. The patients had similar dental protrusion, soft tissue profile measurements, and ages at the posttreatment observation. RESULTS: No significant cephalometric differences between the extraction and nonextraction groups were found at long-term follow-up; both groups showed similar long-term changes. Significant (P < .05) differences were found between males and females at long-term follow-up; male lips became relatively more retrusive, and their profiles became flatter. Significant (P < .05) changes in the profiles were also perceived over time, but there was no relationship between the amount of change perceived and profile changes measured cephalometrically. There were also no significant (P < .05) differences in preferences between orthodontists and laypeople, between extraction and nonextraction patients, or between males and females. CONCLUSIONS: If extraction and nonextraction patients are treated to the same incisor position and lip line, the treatment modality does not affect long-term soft tissue profile changes. Furthermore, the amounts of change perceived by either orthodontists or laypeople were not related to the amount of change measured cephalometrically. PMID- 16214627 TI - Tooth shape preferences in an esthetic smile. AB - INTRODUCTION: The purpose of this study was to evaluate the contributions of tooth shape to the esthetic smile. METHODS: Restorative dentists (120), laypeople (102), and orthodontists (113) evaluated a series of color photographs of men's and women's smiles. The photographs were randomly presented to test the effects of 3 different shapes of maxillary incisors and canines on the same patient. RESULTS: For women, orthodontists preferred round and square-round incisors (P < .01), and restorative dentists preferred round incisors (P < or = .03). Laypeople did not discriminate between incisor shapes. For men, all 3 groups preferred square-round incisors (P < or = .042). There was also a tendency for male judges to rate female images more attractive than did female judges. CONCLUSIONS: Restorative dentists, orthodontists, and laypeople share similarities and display differences when considering esthetic preferences in tooth shape. Although there was no consensus in preference among the laypeople as a group, their preferences differed from those of the dental professionals. PMID- 16214625 TI - Dental esthetics, orthodontic treatment, and oral-health attitudes in young adults. AB - INTRODUCTION: The aim of this study was to investigate whether young adults with varying dental esthetics and histories of orthodontic treatment also differ in oral-health attitudes, preventive behaviors, and self-perceived oral health. METHODS: The sample comprised 298 young adults, 18 to 30 years old, with at least 13 years of primary and secondary school education. The subjects were asked to complete questionnaires dealing with various measures related to oral-health attitudes, preventive behaviors, and perceptions of oral health. Dental esthetics were assessed by means of the aesthetic component of the index of orthodontic treatment need. Dental plaque accumulation was assessed in a subsample of respondents. RESULTS: Subjects with high dental-esthetics scores reported more favorable oral-health attitudes, such as internal control, dental awareness, value of occlusion, and preventive behavior expectations than subjects with lower scores. Subjects with previous orthodontic treatment showed greater internal control and dental awareness than those who had not previously been treated. Subjects ranking high in dental esthetics and those with previous orthodontic treatment reported stricter oral-hygiene adherence than others. Self-perceived oral health was better in high scorers on dental esthetics. Less plaque accumulation was found in subjects with higher dental esthetic scores and in those with previous orthodontic treatment. CONCLUSIONS: These findings suggest that favorable dental esthetics and previous orthodontic treatment might be important variables in explaining individual differences in oral-health attitudes and behaviors. PMID- 16214628 TI - Mandibular prognathism in Japanese families ascertained through orthognathically treated patients. AB - INTRODUCTION: Patients with severe mandibular prognathism frequently require orthognathic surgery to correct skeletal disharmony. Genetic and environmental factors are suggested in the etiology of mandibular prognathism, but the precise contribution of these factors is unknown. METHODS: To clarify the impact of genetic factors on mandibular prognathism, we gave a questionnaire to 105 subjects with severe skeletal Class III mandibular prognathism who planned to undergo or had undergone orthognathic surgery. The questionnaire assessed mandibular prognathism in 3 generations of each subject's family. All answers were confirmed in a careful interview that included differentiation of "maxillary deficiency" and "mandibular excess." This is the first 3-generation family study of mandibular prognathism in orthognathically treated patients. RESULTS: The average ANB, SNA, and SNB angles in the probands were -3.7 degrees , 80.9 degrees , and 84.6 degrees, respectively. A total of 1480 family members were examined, and 11.2% had mandibular prognathism. Men were slightly more affected than women; 68.6% of families had at least 1 member other than the proband with mandibular prognathism. The affected ratio of first-degree relatives was more than twice that of second-degree relatives (17.5% and 7.6%, respectively). There was no difference in the occurrence between fathers and mothers of probands, and siblings showed a high affected ratio of 25.0%. Falconer's heritability was calculated as 84.3% in first-degree relatives. CONCLUSIONS: A high occurrence of mandibular prognathism was seen in families of orthognathically treated skeletal Class III patients, suggesting a profound genetic influence. PMID- 16214629 TI - The effect of stretching on the release of fluoride from fluoridated elastomeric ligatures. AB - INTRODUCTION: The aim of this in-vitro investigation was to determine whether stretching increases the amount of fluoride released from fluoridated elastomeric ligatures. METHODS: Ten groups of 4 fluoridated elastomeric ligatures stretched over edgewise premolar brackets were compared with 10 groups of 4 ligatures that were not stretched over brackets. Each group was placed in individual polyethylene bottles containing 1 mL of distilled water and maintained at 37 degrees C in an incubator. A polyethylene bottle containing only distilled water and 4 brackets was used as the control. The fluoride content of the samples was determined once a day for 7 days and regularly thereafter up to 196 days. A fluoride-ion selective electrode coupled to an analyzer was used to determine the fluoride content of the solutions. The sample solutions were changed 24 hours before readings to prevent the results from being cumulative. RESULTS: The fluoride-ion release for each of the groups was calculated and expressed as total fluoride release in micrograms of fluoride per day per milliliter per elastomer. During the first month in solution, the stretched fluoridated elastomers released approximately 32 microgF.day/mL and the unstretched speciments released 30 microgF.day/mL. This represents a 7% increase in fluoride release, which was statistically significant (P = .007). Over the entire test period (196 days), the modules stretched over the brackets released about 13% more fluoride. This was also a statistically significant difference (P = .001). CONCLUSIONS: Stretching increases the concentration and amount of fluoride released from fluoridated elastomeric ligatures. PMID- 16214630 TI - Effects of prestretching on force degradation of synthetic elastomeric chains. AB - INTRODUCTION: The purpose of this study was to evaluate the effects of prestretching on time-dependant force decay of synthetic elastomeric chains. METHODS: Five-unit (12.5 mm) and 6-unit (15.5 mm) modules (Generation II, Ormco, Glendora, Calif) were prestretched 100% for 1 hour (n = 12), 24 hours (n = 12), 2 weeks (n = 12), and 4 weeks (n = 12) in 37 degrees C distilled water. The prestretched and unprestretched (control) modules were then stretched to 30 mm in 37 degrees C water, and their forces were measured at 0 hour, 1 hour, 24 hours, and weekly for 4 weeks with a digital force gauge. RESULTS: The prestretched 5- and 6-unit modules yielded significantly lower initial force than the controls. All 5- and 6-unit prestretched and control groups showed substantial force decay during the first hour. However, at 1 hour, similar remaining forces were found in the 5-unit prestretched and control groups (P > .05), and small differences were seen in the 6-unit groups. The rates and patterns of force decay from 1 hour to 4 weeks were quite similar between the control and the prestretched modules of both the 5- and 6-unit groups. CONCLUSIONS: The effects of prestretching on force decay of elastomeric chains were noted mainly in the first hour. Thus, the clinical value of prestretching a synthetic elastomeric chain is questionable. PMID- 16214631 TI - Tooth movement and cytokines in gingival crevicular fluid and whole blood in growing and adult subjects. AB - INTRODUCTION: Tooth movement has been studied largely with respect to the force required for tipping when pressure distribution varies along the length of the periodontal ligament. But important factors for effective canine translation include the nature and magnitude of applied stress and the patient's cell biology. The purpose of this research was to test 3 hypotheses: (1) the velocity of tooth translation (v(t)) is related to applied stress and growth status, (2) a threshold of stress accounts for the lag phase, and (3) v(t) is correlated with the ratio (AI) of 2 cytokines (IL-1beta, IL-1RA) measured in gingival crevicular fluid (GCF) and stimulated whole blood (SWB). METHODS: Continuous maxillary canine retraction stresses of 13 kPa and 4, 26, or 52 kPa were applied bilaterally in 6 growing and 4 adult subjects for 84 days. Dental models and GCF samples were collected at 1- to 14-day intervals. Cytokines were measured in GCF and SWB cell cultures. RESULTS: V(t) was positively related to stress and was higher in growing subjects (P = .001). It was also related to AI(GCF) in growers (R2= 0.56) and nongrowers (R2= 0.72). Canines moved with 52 kPa showed a lag phase, and postlag phase AI(GCF) was twice that of lag phase AI(GCF). Mean v(t) and associated AI(GCF) during the postlag phase were nearly double the values for canines moved with 13 and 26 kPa. SWB production of cytokines was dose-dependent. For growing subjects, SWB IL-1RA was correlated with v(t) (R = 0.70-0.72), and AI(SWB) and IL-1beta concentrations were correlated with AI(GCF) (R = 0.73-0.78). CONCLUSIONS: V(t) varied with growth status and stresses < or = 52 kPa; stresses of < 52 kPa showed no lag phase; and equivalent stresses yielded subject dependent differences in v(t), which correlated with cytokines in GCF and SWB. PMID- 16214632 TI - Alkaline phosphatase activity in dental pulp of orthodontically treated teeth. AB - INTRODUCTION: The aim of this study was to examine alkaline phosphatase (ALP) activity in the dental pulp of orthodontically treated teeth. METHODS: Sixteen healthy subjects (mean age 17.0 +/-1.6 years) who required extraction of 4 first premolars for orthodontic reasons participated. One maxillary first premolar subjected to orthodontic force was the test tooth. The contralateral first premolar, bracketed but not subjected to mechanical stress, was the control tooth. After a week of treatment, the first premolars were extracted and the dental pulp removed from the teeth. ALP activity was determined spectrophotometrically and the results expressed as units/liter per milligram of pulp tissue [U/(L x mg)]. RESULTS: ALP activity was 89 +/- 26 U/(L x mg) in the test teeth and 142 +/- 33 U/(L x mg) in the control teeth. The difference between the groups was statistically significant (P < .01). CONCLUSIONS: Orthodontic treatment can lead to significant early-phase reduction in ALP activity in human dental pulp tissue. PMID- 16214633 TI - Expression of mRNA for osteoprotegerin and receptor activator of nuclear factor kappa beta ligand (RANKL) during root resorption induced by the application of heavy orthodontic forces on rat molars. AB - INTRODUCTION: Receptor activator of nuclear factor kappa beta ligand (RANKL) activates osteoclast differentiation, whereas this activity is blocked by osteoprotegrin (OPG), so that the relative expression of these 2 proteins might contribute to bone and root resorption during orthodontic tooth movement. We describe experiments with RANKL and OPG mRNA expression in rats subjected to orthodontic forces. It was hypothesized that the ratios of RANKL to OPG expression would increase during root resorption processes. METHODS: Fixed Sentalloy (GAC, Bohemia, NY) closed-coil springs capable of delivering approximately 100 g of force were applied for mesial movement of the mandibular left first molar in 9 male, 7-week-old Wistar rats; the right mandibular molar was used as an internal control for each animal. After 14 days, the rats were killed; tissues from 2 rats were examined by paraffin histology, and high-quality messenger ribonucleic acid (mRNA) was extracted from 4-mm widths of the mesial bony tissues in the remaining animals. RESULTS: Paraffin sections showed osteoclastic resorption of roots on the mesial surfaces of teeth subjected to orthodontic forces. The integrity of mRNA was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) for the housekeeping gene GAPDH, and that of primers specific for OPG and RANKL was determined by RT-PCR for these genes in material isolated from the UM106 rat cell line known to express both proteins. Densitometric analysis of the RT-PCR OPG product showed an increase in background levels of OPG mRNA in bony tissues subjected to orthodontic forces in all animals studied (P < .05). In contrast, low levels of mRNA for RANKL were detected in only 5 animals and only in association with orthodontic forces. CONCLUSIONS: Data are consistent with changes in levels of OPG and RANKL in tissues subjected to orthodontic forces and experiencing root resorption. PMID- 16214634 TI - The interaction between changes in upright mandibular position and supine airway size in patients with obstructive sleep apnea. AB - INTRODUCTION: The purpose of this study was to investigate the interaction between upright mandibular position change and supine upper airway size in men with obstructive sleep apnea fitted with titratable oral appliances. METHODS: Baseline supine cephalometry before placement of the oral appliance and after titration with the oral appliance in place were undertaken in 14 patients, and upright mandibular position change was evaluated with and without the titrated oral appliance in place with a DigiGraph workstation (Dolphin Imaging Systems, Valencia, Calif). RESULTS: The apnea-hypopnea index was significantly reduced after titration of the oral appliance (P < .01). Upright mandibular position change was associated with significant vertical (P < .01) and horizontal (P < .01) mandibular repositioning. The size of the supine velopharynx (P < .05), but not the supine oropharynx, was significantly enlarged at the titrated mandibular position. The supine oropharyngeal size change was correlated with the upright horizontal repositioning of the mandible (r = 0.69, P < .01). CONCLUSIONS: Evaluation of upright mandibular position changes with the DigiGraph workstation enables one to predict supine oropharyngeal enlargement with oral appliance therapy. Dose-dependent effects of the horizontal component of upright mandibular protrusion on supine oropharyngeal size in addition to velopharyngeal enlargement might contribute to oral appliance effectiveness in obstructive sleep apnea patients. PMID- 16214635 TI - Reproducibility of airway dimensions and tongue and hyoid positions on lateral cephalograms. AB - INTRODUCTION: The aim of this study was to evaluate the reproducibility of airway dimensions and tongue and hyoid positions on lateral cephalometric radiographs. METHODS: Three lateral cephalograms each of 30 patients were obtained in natural head positions at 30-minute intervals. Twelve measurements, including pharyngeal airway dimensions and tongue and hyoid positions, were taken. The relationships between 3 sets of measurements were evaluated by using repeated analysis of variance, Dahlberg's method error formula, and correlation coefficient. RESULTS: No statistically significant differences were found between the 3 sets of measurements with the repeated analysis of variance (P > .05). Correlation coefficient values ranged between 0.964 (vertical position of the hyoid) and 0.683 (hypopharyngeal airway width). The average method error was 1.22 mm. CONCLUSIONS: The results suggest that airway dimension and tongue- and hyoid position measurements are highly reproducible on natural-head-position cephalograms. PMID- 16214636 TI - Sucking habits and facial hyperdivergency as risk factors for anterior open bite in the mixed dentition. AB - INTRODUCTION: The aim of this study was to evaluate sucking habits and hyperdivergency as risk factors for anterior open bite in mixed-dentition subjects. METHODS: Anamnestic and pretreatment cephalometric records of 1710 mixed-dentition subjects were assessed for sucking habits, dental open bite, and facial hyperdivergency. RESULTS: The rate of anterior open bite was 17.7%. Multiple logistic regression showed that both prolonged sucking habits and hyperdivergent vertical relationships significantly increased the probability of an anterior dentoalveolar open bite, with a prevalence rate of 36.3%. This was 4 times the prevalence of sucking habits and facial hyperdivergency in subjects without anterior open bite (9.1%). CONCLUSIONS: Prolonged sucking habits and hyperdivergent facial characteristics are significant risk factors for anterior open bite in the mixed dentition. PMID- 16214637 TI - Two-stage treatment of skeletal Class III malocclusion during the early permanent dentition. AB - A patient with skeletal Class III malocclusion was treated in 2 phases during the early permanent dentition. In phase 1, maxillary protraction was combined with rapid palatal expansion; in phase 2, fixed appliances were placed. The results were good posttreatment, and, 1 year later, a favorable growth tendency could be observed. This report shows that treatment for a patient with skeletal Class III malocclusion can be started in the early permanent dentition, with very good final results. PMID- 16214638 TI - Category 7: Class II skeletal malocclusion with transverse maxillary constriction in an adult patient. AB - This case report was submitted to the American Board of Orthodontics as part of the board-certification process. The summary of treatment and records are reprinted here much as they were submitted to the board. PMID- 16214639 TI - Orthodontics in 3 millennia. Chapter 5: the American Board of Orthodontics, Albert Ketcham, and early 20th-century appliances. AB - Early in the last century, 3 events put Colorado in the orthodontic spotlight: the discovery-by an orthodontist-of the caries-preventive powers of fluoridated water, the formation of dentistry's first specialty board, and the founding of a supply company by and for orthodontists. Meanwhile, inventive practitioners were giving the profession more choices of treatment modalities, and stainless steel was making its feeble debut. PMID- 16214640 TI - The new American Board of Orthodontics certification process: further clarification. PMID- 16214641 TI - Litigation, legislation, and ethics. Are you a doctor ... ? PMID- 16214643 TI - Sickle cell vaso-occlusion. AB - The vaso-occlusion model has evolved impressively over the past several decades from polymerization-based concepts to a complex, wide-ranging schema that involves multistep, heterogeneous, and interdependent interactions among sickle erythrocytes (SSRBCs), adherent leukocytes, endothelial cells, plasma proteins, and other factors. Endothelial activation, induced directly or indirectly by the proinflammatory behavior of SSRBCs, is the most likely initiating step toward vaso-occlusion. Given the complexity and dynamic relationships of the potential mechanisms leading to vaso-occlusion, further in vivo studies in relevant sickle cell animal models will most likely yield the greatest advances and promote the development of novel, more effective therapeutic strategies. PMID- 16214644 TI - Pain management of sickle cell disease. AB - Management of sickle cell disease continues to be primarily palliative, including supportive, symptomatic, and preventive approaches to therapy. There are three major types of sickle cell pain: acute, chronic, and neuropathic pain. The acute painful episode is the insignia of the disease and the most common cause of hospitalization. Its management entails the use of nonpharmacologic and pharmacologic modalities. Pain management should follow certain principles that include an assessment stage, treatment stage, reassessment stage, and adjustment stage. Chronic sickle cell pain may be due to certain complications of the disease, such as leg ulcers and avascular necrosis; intractable chronic pain may be due to central sensitization. Management of chronic pain should take a multidisciplinary approach. PMID- 16214645 TI - Transfusion management in sickle cell disease. AB - Sickle cell disease (SCD) is the most commonly inherited hemoglobinopathy in the United States. Blood transfusion is a critical part of the multidisciplinary approach necessary in the management of SCD; however, blood transfusions are not without complications. The successful use of transfusion as a treatment strategy in SCD requires the critical review and knowledge of transfusion methods, generally accepted indications, clinical situations in which transfusion generally is not considered, the selection of blood products, and strategies to prevent transfusion-related complications. PMID- 16214646 TI - Arterial blood pressure and hyperviscosity in sickle cell disease. AB - Abnormal rheologic behavior of sickle cells is the result of increased viscosity of the blood caused by the polymerization of hemoglobin S and the resultant production of dense, dehydrated sickle erythrocytes. As the viscosity of sickle cells increases, there is a negative impact on blood flow, which contributes to the vascular occlusion process, the hallmark of the sickling disorders. Blood flow is directly proportional to the blood pressure and inversely proportional to the blood viscosity. Blood flow has important implications for the diagnosis and management of hypertension in sickle cell patients and for transfusion therapy for the acute and chronic complications of this disease. PMID- 16214647 TI - Overt and incomplete (silent) cerebral infarction in sickle cell anemia: diagnosis and management. AB - Regional complete infarctions in children with sickle cell anemia (HbSS) are often associated with stenosis of the large intracranial arteries and result in lifetime disability. Incomplete infarction occurs more frequently than previously recognized and has far-reaching effects on neurocognitive development and the risk for overt secondary strokes into adulthood. Clinical and neuroimaging modalities have been highlighted in an algorithmic approach, with the studies giving the highest yield in results and most likely to be available listed in sequential order. The recognition of an emerging "second peak" incidence in the third decade of life is worrisome and warrants more intense scrutiny and diagnosis of subtle findings of stroke in this young adult population. PMID- 16214648 TI - The acute chest syndrome. AB - Recent large clinical studies of the acute chest syndrome (ACS) have improved our understanding of its pathophysiology and epidemiology. However, there is still a need for better methods of distinguishing vaso-occlusion from fibrin or fat embolism, for rapid diagnostic tests to make positive identifications of microbial infection, for adjunctive therapies that would affect prognosis, and for identification of factors that influence prognosis. The difference in clinical course and severity between children and adults supports the results of current studies indicating multiple causes for ACS. The mainstay of successful treatment remains high-quality supportive care. The judicious use of transfusion therapy has a major role in preventing mortality in the absence of a specific therapy that consistently improves the clinical course. PMID- 16214649 TI - Pulmonary hypertension in sickle cell disease: mechanisms, diagnosis, and management. AB - Pulmonary hypertension affects nearly one-third of adults with sickle cell disease and is related to hemolysis. Although pulmonary pressures are not as high as in other forms of pulmonary hypertension, sickle cell patients poorly tolerate even moderate elevations of pulmonary pressures, because this complication predicts short survival. Tricuspid regurgitant jet velocity measured noninvasively by echocardiography is an adequate tool to screen for pulmonary hypertension. Because patients with pulmonary hypertension are older and often severely affected by other vaso-occlusive complications, optimizing their treatment with hydroxyurea or transfusions is important. Newer agents such as sildenafil and bosentan have demonstrated efficacy in other forms of pulmonary hypertension, and future clinical trials may prove them helpful in sickle cell patients. PMID- 16214650 TI - Surgery in sickle cell disease. AB - Persons with sickle cell disease (SCD) are more likely to undergo surgery than are the general population during their lifetime. For example, cholecystectomy as a consequence of gallstones is more frequent in persons with SCD, as is hip arthroplasty in younger people as a result of avascular necrosis of the femoral head. Because surgery exposes patients to many of the factors that are known to precipitate red blood cell sickling, persons with SCD undergoing surgery require meticulous clinical care to prevent perioperative sickle cell-related complications. Even with meticulous care, approximately 25% to 30% of patients will have a postoperative complication. This article provides readers with information about the role of surgery in SCD and the measures that should be taken to ensure patients are well cared for in the perioperative period. PMID- 16214651 TI - Pregnancy and sickle cell disease. AB - With advances in management, men and women with sickle cell disease are enjoying an improved quality of life well into adulthood, when they may elect to plan a family. Pregnancy has been associated with exacerbation of sickle cell disease and may place women, especially those with sickle cell anemia (HbSS), at an additional risk for obstetric complications. Appropriate management by health care providers familiar with sickle cell diseases and high-risk obstetric care can result in a successful pregnancy for most women with sickle cell disease. PMID- 16214652 TI - Priapism in sickle cell disease. AB - Priapism, an unwanted painful erection of the penis, is a little discussed but common complication of sickle cell disease. What is known about the prevalence of priapism, efficacy of management approaches, and outcome is drawn primarily from retrospective and single-center reports. Priapism occurs in two patterns: prolonged and stuttering (ie, recurrent brief episodes that resolve spontaneously). If priapism persists for 4 hours or more without detumescence, the patient is at risk for irreversible ischemic penile injury, which may terminate in fibrosis and impotence. Large multicenter studies examining the epidemiology and current treatments and well-organized trials of novel therapies are urgently needed for patients who have sickle cell disease and priapism. PMID- 16214653 TI - Bone and joint disease in sickle cell disease. AB - Bone and joint disorders are the most common cause of chronic pain in patients who have sickle cell disease. The femoral head is the most common area of bone destruction in sickle cell patients, although other disease-related problems include avascular necrosis of the humeral head, changes in the thoracic and lumbar spine, infection with encapsulated organisms (Salmonella and Staphylococcus aureus are the most common), bone marrow disturbances, and dental effects. Complications can occur at any location: epiphyseal, metaphyseal, or diaphyseal. The location and the extensiveness of the problems determine the pain and structural damage. The hip joint is particularly vulnerable in sickle cell disease. This article highlights aspects of sickle cell disease that affect healthy bone and joint function and discusses treatment options. PMID- 16214654 TI - Leg ulceration in sickle cell disease: medieval medicine in a modern world. AB - Leg ulceration is now recognized as an important complication of sickle cell disease, especially of the SS genotype. Since there is no convincing evidence of delayed healing of operation scars or of wounds elsewhere in the body, it must be concluded that factors specific to the lower leg render patients prone to delayed healing at this site. Many lesions are traumatic in origin and since there is considerable variation in healing rates among the normal population, it is useful to define chronic leg ulceration on the basis of a minimal duration, which in Jamaican studies has required at least 3 months and sometimes 6 months before healing. This minimal duration avoids the difficulties of interpreting the significance of briefer lesions since the moment of final healing may be poorly defined (patients may conclude that a scab represents healing whereas small lesions persist beneath) and often goes undocumented as patients may not report and medical attendants may not enquire, the date of final healing. PMID- 16214655 TI - Effects of sickle cell disease on the eye: clinical features and treatment. AB - Ocular complications of the sickling disorders are multiple and require continuing assessment to detect lesions early enough for effective prophylactic therapy to maximize visual functioning. The disease requires close monitoring of the patient for progression or regression of lesions in determining the selection of therapeutic options. PMID- 16214656 TI - New therapies for sickle cell disease. AB - New and developing therapeutic agents for the treatment of sickle cell disease include hydroxyurea (an unlicensed experimental drug in most countries), omega-3 fatty acids, and the Gardos channel inhibitor ICA-17043. Anti-cellular adhesion therapy has considerable prospects; however, it has yet to be translated into clinical practice. For specific disease manifestations, pulmonary hypertension responds well to oral arginine, l-carnitine, and exchange blood transfusion therapy alone or in combination with other agents. Primary stroke prevention with transfusion therapy is now considered standard care. Oral iron chelators are administered increasingly instead of the more inconvenient parenteral desferrioxamine. Deferiprone is licensed in Europe and India, and deferasirox (ICL670) holds out important promise because it has not been shown to affect blood cell counts. PMID- 16214657 TI - Assessment of two-dimensional induced accelerations from measured kinematic and kinetic data. AB - A simple algorithm is presented to calculate the induced accelerations of body segments in human walking for the sagittal plane. The method essentially consists of setting up 2x4 force equations, 4 moment equations, 2x3 joint constraint equations and two constraints related to the foot-ground interaction. Data needed for the equations are, next to masses and moments of inertia, the positions of ankle, knee and hip. This set of equations is put in the form of an 18x18 matrix or 20x20 matrix, the solution of which can be found by inversion. By applying input vectors related to gravity, to centripetal accelerations or to muscle moments, the 'induced' accelerations and reaction forces related to these inputs can be found separately. The method was tested for walking in one subject. Good agreement was found with published results obtained by much more complicated three-dimensional forward dynamic models. PMID- 16214658 TI - Biomechanical and electromyographic evaluation of ankle foot orthosis and dynamic ankle foot orthosis in spastic cerebral palsy. AB - This study evaluated the biomechanical and electromyographic effects of conventional ankle foot orthoses (AFOs) and dynamic ankle foot orthoses (DAFOs) on gait in patients with spastic cerebral palsy (CP). Thirteen patients with dynamic equinus underwent motion analysis with electromyography. Both AFOs and DAFOs provided longer stride length, permitted pre-positioning for initial contact, and successfully controlled the excessive plantarflexion during the swing phase. Median frequency (MF) of EMG signal indicated that extremely high firing was found in the patient's lower limbs compared to controls that resulted in tiredness. The DAFOs allowed a significantly larger total ankle range of motion than the AFOs. However, AFOs significantly reduced the MF while DAFOs did not. The reduced MF seen when wearing AFOs suggested an improvement of walking endurance. The DAFO had the advantage of less restriction on ankle movement, which avoids muscular atrophy and improves orthotic compliance. PMID- 16214659 TI - Compensatory stepping in response to waist pulls in balance-impaired and unimpaired women. AB - An effective stepping response is often critical in avoiding a fall. Our objective was to study the effects of age and balance impairment on anterior and posterior compensatory stepping strategies in response to waist pull perturbations of 1-5% body weight (BW). Based on maximal unipedal stance time (UST), we tested 15 balance-impaired old (BI, UST < 10s, mean age = 76 years), 12 healthy old (O, UST>30s, mean age = 71 years), and 13 healthy young women (Y, UST >30s, mean age=23 years). Randomized anterior and posterior pulls of 1-5% body weight (BW) were applied to the waist while kinematic and kinetic recovery responses were recorded. Results show that O and BI required 0.5 more steps than Y to recover balance for posterior pulls of 4-5% BW (P < 0.01). For anterior pulls of 4-5% BW, only BI had a greater probability of step initiation (P4 weeks with either efavirenz (EFV) or protease inhibitors (PIs) as part of their antiretroviral therapy (ART). METHOD: A cross-sectional, questionnaire-based cohort of HIV patients who received two nucleoside reverse transcriptase inhibitors combined with either EFV (n = 75) or one or more PIs (n = 77) for at least 4 weeks and were tolerating therapy. The extent of neuropsychiatric disturbances was evaluated based on self reported symptoms using the psychological evaluation test SCL-90-R. Treatment duration was broken down into quartiles of 30-198 days, >198-365 days, >365-637 days, and >637 days. RESULTS: In the first 6 months of therapy, significantly higher (worse) scores were seen in 39/75 (52%) of the patients receiving EFV. The EFV-treated group had significantly higher scores for somatization, anxiety, obsessive-compulsive behavior, the Global Severity Index, and the Positive Symptom Distress Index, with trends for higher scores in paranoid ideation and depression symptom subcategories. Over the following 6 to 12 months of therapy, the EFV group scored higher than the PI group in somatization, anxiety, obsessive compulsive behavior, hostility, depression, the Global Severity Index, and the Positive Symptom Total, but the differences were not significant. After 12 months of therapy, the EFV group had significantly lower scores than the PI group for somatization, interpersonal sensitivity, Global Severity Index, and Positive Symptom Total. CONCLUSION: EFV-induced neuropsychiatric symptoms can last up to 200 days after treatment initiation. However, symptom severity appears to decline over time in EFV-treated patients versus patients treated with a PI-based ART. PMID- 16214736 TI - Reductions in stavudine dose might ameliorate mitochondrial-associated complications without compromising antiviral activity. AB - BACKGROUND: Stavudine (d4T) is a nucleoside analogue approved for the treatment of HIV infection. Concern has risen due to its association with mitochondrial toxicity. Given that the toxicity might be dose-dependent, we explored prospectively whether lowering d4T doses might improve the safety profile of the drug without compromising its antiviral activity. METHOD: All HIV-infected patients seen at our institution during the first semester of year 2003 who were receiving a d4T-containing regimen and had plasma HIV RNA below 50 copies/mL for the previous 3 months were invited to participate in a trial in which half of patients reduced the dose of d4T from 40 to 30 mg bid (cases) and the other half continued with the same d4T dose (controls). RESULTS: A total of 92 patients were recruited in the study: 47 cases and 45 controls. A total of 9 patients experienced virological failure during the following 12 months: 4 cases and 5 controls. No significant differences between groups were recognized for mean transaminase levels, cholesterol, triglycerides, and lactate at baseline nor over the 12-month follow-up period. Lipodystrophy was recognized in 20% of patients at baseline, without significant differences between groups, and no significant improvements were recognized in the d4T 30 mg bid arm after 12 months follow-up. However, a median significant increase of 2.23-fold in the mitochondrial DNA content in peripheral blood mononuclear cells (PBMCs) was recognized in a subset of 11 patients who reduced the d4T dose, whereas it remained unchanged in 10 controls. CONCLUSION: A reduction in the d4T dose from 40 to 30 mg bid may ameliorate mtDNA depletion in PBMCs without compromising the antiviral activity of the drug. However, significant improvements on surrogate laboratory markers of mitochondrial toxicity or in lipoatrophy could not be recognized over 12 months follow-up. PMID- 16214737 TI - Bacterial infections in adult HIV-infected patients. AB - PURPOSE: The purpose of this article is to critically review articles published from the pre-HAART era to the present on bacterial infections in adult HIV infected patients. METHOD: Literature search on bacterial infections in HIV infected patients yields predominantly small case series from single centers, many of which are retrospectively collected. RESULTS: Variations in case selection limit the utility of these articles for assessing the epidemiology and clinical features of a particular infection. Nonetheless, numerous articles indicate that certain bacterial infections occur most often when CD4 cell counts are < 200/mm3. In the pre-HAART era, others suggest that PcP prophylaxis with TMP/SMX and MAC prophylaxis with macrolides reduced rates of several bacterial infections. Since the advent of HAART, however, some articles suggest that the incidence of various infections has declined and that withdrawal of OI prophylaxis in patients who have had HAART restoration of CD4 cell counts has not led to an increase in certain bacterial infections. CONCLUSION: This review suggests that bacterial infections may have declined in the HAART era, as multicenter cohort studies have shown to be the case with AIDS-associated OIs. Nonetheless, preventive measures such as pneumococcal vaccination and smoking cessation remain effective strategies. PMID- 16214738 TI - Report from Boston: the 12th Conference on Retroviruses and Opportunistic Infections (CROI). February 22-25, 2005. Complications/toxicity of HIV therapy. PMID- 16214740 TI - DNA barcodes for biosecurity: invasive species identification. AB - Biosecurity encompasses protecting against any risk through 'biological harm', not least being the economic impact from the spread of pest insects. Molecular diagnostic tools provide valuable support for the rapid and accurate identification of morphologically indistinct alien species. However, these tools currently lack standardization. They are not conducive to adaptation by multiple sectors or countries, or to coping with changing pest priorities. The data presented here identifies DNA barcodes as a very promising opportunity to address this. DNA of tussock moth and fruit fly specimens intercepted at the New Zealand border over the last decade were reanalysed using the cox1 sequence barcode approach. Species identifications were compared with the historical dataset obtained by PCR-RFLP of nuclear rDNA. There was 90 and 96% agreement between the methods for these species, respectively. Improvements included previous tussock moth 'unknowns' being placed to family, genera or species and further resolution within fruit fly species complexes. The analyses highlight several advantages of DNA barcodes, especially their adaptability and predictive value. This approach is a realistic platform on which to build a much more flexible system, with the potential to be adopted globally for the rapid and accurate identification of invasive alien species. PMID- 16214739 TI - Towards writing the encyclopedia of life: an introduction to DNA barcoding. AB - An international consortium of major natural history museums, herbaria and other organizations has launched an ambitious project, the 'Barcode of Life Initiative', to promote a process enabling the rapid and inexpensive identification of the estimated 10 million species on Earth. DNA barcoding is a diagnostic technique in which short DNA sequence(s) can be used for species identification. The first international scientific conference on Barcoding of Life was held at the Natural History Museum in London in February 2005, and here we review the scientific challenges discussed during this conference and in previous publications. Although still controversial, the scientific benefits of DNA barcoding include: (i) enabling species identification, including any life stage or fragment, (ii) facilitating species discoveries based on cluster analyses of gene sequences (e.g. cox1 = CO1, in animals), (iii) promoting development of handheld DNA sequencing technology that can be applied in the field for biodiversity inventories and (iv) providing insight into the diversity of life. PMID- 16214741 TI - DNA barcoding for effective biodiversity assessment of a hyperdiverse arthropod group: the ants of Madagascar. AB - The role of DNA barcoding as a tool to accelerate the inventory and analysis of diversity for hyperdiverse arthropods is tested using ants in Madagascar. We demonstrate how DNA barcoding helps address the failure of current inventory methods to rapidly respond to pressing biodiversity needs, specifically in the assessment of richness and turnover across landscapes with hyperdiverse taxa. In a comparison of inventories at four localities in northern Madagascar, patterns of richness were not significantly different when richness was determined using morphological taxonomy (morphospecies) or sequence divergence thresholds (Molecular Operational Taxonomic Unit(s); MOTU). However, sequence-based methods tended to yield greater richness and significantly lower indices of similarity than morphological taxonomy. MOTU determined using our molecular technique were a remarkably local phenomenon-indicative of highly restricted dispersal and/or long term isolation. In cases where molecular and morphological methods differed in their assignment of individuals to categories, the morphological estimate was always more conservative than the molecular estimate. In those cases where morphospecies descriptions collapsed distinct molecular groups, sequence divergences of 16% (on average) were contained within the same morphospecies. Such high divergences highlight taxa for further detailed genetic, morphological, life history, and behavioral studies. PMID- 16214742 TI - Wedding biodiversity inventory of a large and complex Lepidoptera fauna with DNA barcoding. AB - By facilitating bioliteracy, DNA barcoding has the potential to improve the way the world relates to wild biodiversity. Here we describe the early stages of the use of cox1 barcoding to supplement and strengthen the taxonomic platform underpinning the inventory of thousands of sympatric species of caterpillars in tropical dry forest, cloud forest and rain forest in northwestern Costa Rica. The results show that barcoding a biologically complex biota unambiguously distinguishes among 97% of more than 1000 species of reared Lepidoptera. Those few species whose barcodes overlap are closely related and not confused with other species. Barcoding also has revealed a substantial number of cryptic species among morphologically defined species, associated sexes, and reinforced identification of species that are difficult to distinguish morphologically. For barcoding to achieve its full potential, (i) ability to rapidly and cheaply barcode older museum specimens is urgent, (ii) museums need to address the opportunity and responsibility for housing large numbers of barcode voucher specimens, (iii) substantial resources need be mustered to support the taxonomic side of the partnership with barcoding, and (iv) hand-held field-friendly barcorder must emerge as a mutualism with the taxasphere and the barcoding initiative, in a manner such that its use generates a resource base for the taxonomic process as well as a tool for the user. PMID- 16214743 TI - DNA barcoding Australia's fish species. AB - Two hundred and seven species of fish, mostly Australian marine fish, were sequenced (barcoded) for a 655 bp region of the mitochondrial cytochrome oxidase subunit I gene (cox1). Most species were represented by multiple specimens, and 754 sequences were generated. The GC content of the 143 species of teleosts was higher than the 61 species of sharks and rays (47.1% versus 42.2%), largely due to a higher GC content of codon position 3 in the former (41.1% versus 29.9%). Rays had higher GC than sharks (44.7% versus 41.0%), again largely due to higher GC in the 3rd codon position in the former (36.3% versus 26.8%). Average within species, genus, family, order and class Kimura two parameter (K2P) distances were 0.39%, 9.93%, 15.46%, 22.18% and 23.27%, respectively. All species could be differentiated by their cox1 sequence, although single individuals of each of two species had haplotypes characteristic of a congener. Although DNA barcoding aims to develop species identification systems, some phylogenetic signal was apparent in the data. In the neighbour-joining tree for all 754 sequences, four major clusters were apparent: chimaerids, rays, sharks and teleosts. Species within genera invariably clustered, and generally so did genera within families. Three taxonomic groups-dogfishes of the genus Squalus, flatheads of the family Platycephalidae, and tunas of the genus Thunnus-were examined more closely. The clades revealed after bootstrapping generally corresponded well with expectations. Individuals from operational taxonomic units designated as Squalus species B through F formed individual clades, supporting morphological evidence for each of these being separate species. We conclude that cox1 sequencing, or 'barcoding', can be used to identify fish species. PMID- 16214744 TI - The problems and promise of DNA barcodes for species diagnosis of primate biomaterials. AB - The Integrated Primate Biomaterials and Information Resource (www.IPBIR.org) provides essential research reagents to the scientific community by establishing, verifying, maintaining, and distributing DNA and RNA derived from primate cell cultures. The IPBIR uses mitochondrial cytochrome c oxidase subunit I sequences to verify the identity of samples for quality control purposes in the accession, cell culture, DNA extraction processes and prior to shipping to end users. As a result, IPBIR is accumulating a database of 'DNA barcodes' for many species of primates. However, this quality control process is complicated by taxon specific patterns of 'universal primer' failure, as well as the amplification or co amplification of nuclear pseudogenes of mitochondrial origins. To overcome these difficulties, taxon specific primers have been developed, and reverse transcriptase PCR is utilized to exclude these extraneous sequences from amplification. DNA barcoding of primates has applications to conservation and law enforcement. Depositing barcode sequences in a public database, along with primer sequences, trace files and associated quality scores, makes this species identification technique widely accessible. Reference DNA barcode sequences should be derived from, and linked to, specimens of known provenance in web accessible collections in order to validate this system of molecular diagnostics. PMID- 16214745 TI - Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications. AB - Marine macroalgae, especially the Rhodophyta, can be notoriously difficult to identify owing to their relatively simple morphology and anatomy, convergence, rampant phenotypic plasticity, and alternation of heteromorphic generations. It is thus not surprising that algal systematists have come to rely heavily on genetic tools for molecular assisted alpha taxonomy. Unfortunately the number of suitable marker systems in the three available genomes is enormous and, although most workers have settled on one of three or four models, the lack of an accepted standard hinders the comparison of results between laboratories. The advantages of a standard system are obvious for practical purposes of species discovery and identification; as well, compliance with a universal marker, such as cox1 being developed under the label 'DNA barcode', would allow algal systematists to benefit from the rapidly emerging technologies. Novel primers were developed for red algae to PCR amplify and sequence the 5' cox1 'barcode' region and were used to assess three known species-complex questions: (i) Mazzaella species in the Northeast Pacific; (ii) species of the genera Dilsea and Neodilsea in the Northeast Pacific; and (iii) Asteromenia peltata from three oceans. These models were selected because they have all caused confusion with regards to species number, distribution, and identification in the field, and because they have all been studied with molecular tools. In all cases the DNA barcode resolved accurately and unequivocally species identities and, with the enhanced sampling here, turned up a variety of novel observations in need of further taxonomic investigation. PMID- 16214746 TI - Land plants and DNA barcodes: short-term and long-term goals. AB - Land plants have had the reputation of being problematic for DNA barcoding for two general reasons: (i) the standard DNA regions used in algae, animals and fungi have exceedingly low levels of variability and (ii) the typically used land plant plastid phylogenetic markers (e.g. rbcL, trnL-F, etc.) appear to have too little variation. However, no one has assessed how well current phylogenetic resources might work in the context of identification (versus phylogeny reconstruction). In this paper, we make such an assessment, particularly with two of the markers commonly sequenced in land plant phylogenetic studies, plastid rbcL and internal transcribed spacers of the large subunits of nuclear ribosomal DNA (ITS), and find that both of these DNA regions perform well even though the data currently available in GenBank/EBI were not produced to be used as barcodes and BLAST searches are not an ideal tool for this purpose. These results bode well for the use of even more variable regions of plastid DNA (such as, for example, psbA-trnH) as barcodes, once they have been widely sequenced. In the short term, efforts to bring land plant barcoding up to the standards being used now in other organisms should make swift progress. There are two categories of DNA barcode users, scientists in fields other than taxonomy and taxonomists. For the former, the use of mitochondrial and plastid DNA, the two most easily assessed genomes, is at least in the short term a useful tool that permits them to get on with their studies, which depend on knowing roughly which species or species groups they are dealing with, but these same DNA regions have important drawbacks for use in taxonomic studies (i.e. studies designed to elucidate species limits). For these purposes, DNA markers from uniparentally (usually maternally) inherited genomes can only provide half of the story required to improve taxonomic standards being used in DNA barcoding. In the long term, we will need to develop more sophisticated barcoding tools, which would be multiple, low-copy nuclear markers with sufficient genetic variability and PCR-reliability; these would permit the detection of hybrids and permit researchers to identify the 'genetic gaps' that are useful in assessing species limits. PMID- 16214747 TI - Microcoding: the second step in DNA barcoding. AB - After the process of DNA barcoding has become well advanced in a group of organisms, as it has in the economically important fungi, the question then arises as to whether shorter and literally more barcode-like DNA segments should be utilized to facilitate rapid identification and, where applicable, detection. Through appropriate software analysis of typical full-length barcodes (generally over 500 base pairs long), uniquely distinctive oligonucleotide 'microcodes' of less than 25 bp can be found that allow rapid identification of circa 100-200 species on various array-like platforms. Microarrays can in principle fulfill the function of microcode-based species identification but, because of their high cost and low level of reusability, they tend to be less cost-effective. Two alternative platforms in current use in fungal identification are reusable nylon based macroarrays and the Luminex system of specific, colour-coded DNA detection beads analysed by means of a flow cytometer. When the most efficient means of rapid barcode-based species identification is sought, a choice can be made either for one of these methodologies or for basic high-throughput sequencing, depending on the strategic outlook of the investigator and on current costs. Arrays and functionally similar platforms may have a particular advantage when a biologically complex material such as soil or a human respiratory secretion sample is analysed to give a census of relevant species present. PMID- 16214749 TI - Reverse taxonomy: an approach towards determining the diversity of meiobenthic organisms based on ribosomal RNA signature sequences. AB - Organisms living in or on the sediment layer of water bodies constitute the benthos fauna, which is known to harbour a large number of species of diverse taxonomic groups. The benthos plays a significant role in the nutrient cycle and it is, therefore, of high ecological relevance. Here, we have explored a DNA taxonomic approach to access the meiobenthic organismic diversity, by focusing on obtaining signature sequences from a part of the large ribosomal subunit rRNA (28S), the D3-D5 region. To obtain a broad representation of taxa, benthos samples were taken from 12 lakes in Germany, representing different ecological conditions. In a first approach, we have extracted whole DNA from these samples, amplified the respective fragment by PCR, cloned the fragments and sequenced individual clones. However, we found a relatively large number of recombinant clones that must be considered PCR artefacts. In a second approach we have, therefore, directly sequenced PCR fragments that were obtained from DNA extracts of randomly picked individual organisms. In total, we have obtained 264 new unique sequences, which can be readily placed into taxon groups, based on phylogenetic comparison with currently available database sequences. The group with the highest taxon abundance were nematodes and protozoa, followed by chironomids. However, we find also that we have by far not exhausted the diversity of organisms in the samples. Still, our data provide a framework within which a meiobenthos DNA signature sequence database can be constructed, that will allow to develop the necessary techniques for studying taxon diversity in the context of ecological analysis. Since many taxa in our analysis are initially only identified via their signature sequences, but not yet their morphology, we propose to call this approach 'reverse taxonomy'. PMID- 16214748 TI - The unholy trinity: taxonomy, species delimitation and DNA barcoding. AB - Recent excitement over the development of an initiative to generate DNA sequences for all named species on the planet has in our opinion generated two major areas of contention as to how this 'DNA barcoding' initiative should proceed. It is critical that these two issues are clarified and resolved, before the use of DNA as a tool for taxonomy and species delimitation can be universalized. The first issue concerns how DNA data are to be used in the context of this initiative; this is the DNA barcode reader problem (or barcoder problem). Currently, many of the published studies under this initiative have used tree building methods and more precisely distance approaches to the construction of the trees that are used to place certain DNA sequences into a taxonomic context. The second problem involves the reaction of the taxonomic community to the directives of the 'DNA barcoding' initiative. This issue is extremely important in that the classical taxonomic approach and the DNA approach will need to be reconciled in order for the 'DNA barcoding' initiative to proceed with any kind of community acceptance. In fact, we feel that DNA barcoding is a misnomer. Our preference is for the title of the London meetings--Barcoding Life. In this paper we discuss these two concerns generated around the DNA barcoding initiative and attempt to present a phylogenetic systematic framework for an improved barcoder as well as a taxonomic framework for interweaving classical taxonomy with the goals of 'DNA barcoding'. PMID- 16214750 TI - DNA-based species delineation in tropical beetles using mitochondrial and nuclear markers. AB - DNA barcoding has been successfully implemented in the identification of previously described species, and in the process has revealed several cryptic species. It has been noted that such methods could also greatly assist in the discovery and delineation of undescribed species in poorly studied groups, although to date the feasibility of such an approach has not been examined explicitly. Here, we investigate the possibility of using short mitochondrial and nuclear DNA sequences to delimit putative species in groups lacking an existing taxonomic framework. We focussed on poorly known tropical water beetles (Coleoptera: Dytiscidae, Hydrophilidae) from Madagascar and dung beetles (Scarabaeidae) in the genus Canthon from the Neotropics. Mitochondrial DNA sequence variation proved to be highly structured, with >95% of the observed variation existing between discrete sets of very closely related genotypes. Sequence variation in nuclear 28S rRNA among the same individuals was lower by at least an order of magnitude, but 16 different genotypes were found in water beetles and 12 genotypes in Canthon, differing from each other by a minimum of two base pairs. The distribution of these 28S rRNA genotypes in individuals exactly matched the distribution of mtDNA clusters, suggesting that mtDNA patterns were not misleading because of introgression. Moreover, in a few cases where sequence information was available in GenBank for morphologically defined species of Canthon, these matched some of the DNA-based clusters. These findings demonstrate that clusters of close relatives can be identified readily in the sequence variation obtained in field collected samples, and that these clusters are likely to correspond to either previously described or unknown species. The results suggest that DNA-assisted taxonomy will not require more than a short fragment of mtDNA to provide a largely accurate picture of species boundaries in these groups. Applied on a large scale, this DNA-based approach could greatly improve the rate of species discovery in the large assemblages of insects that remain undescribed. PMID- 16214751 TI - Defining operational taxonomic units using DNA barcode data. AB - The scale of diversity of life on this planet is a significant challenge for any scientific programme hoping to produce a complete catalogue, whatever means is used. For DNA barcoding studies, this difficulty is compounded by the realization that any chosen barcode sequence is not the gene 'for' speciation and that taxa have evolutionary histories. How are we to disentangle the confounding effects of reticulate population genetic processes? Using the DNA barcode data from meiofaunal surveys, here we discuss the benefits of treating the taxa defined by barcodes without reference to their correspondence to 'species', and suggest that using this non-idealist approach facilitates access to taxon groups that are not accessible to other methods of enumeration and classification. Major issues remain, in particular the methodologies for taxon discrimination in DNA barcode data. PMID- 16214752 TI - An integrated approach to fast and informative morphological vouchering of nematodes for applications in molecular barcoding. AB - Molecular surveys of meiofaunal diversity face some interesting methodological challenges when it comes to interstitial nematodes from soils and sediments. Morphology-based surveys are greatly limited in processing speed, while barcoding approaches for nematodes are hampered by difficulties of matching sequence data with traditional taxonomy. Intermediate technology is needed to bridge the gap between both approaches. An example of such technology is video capture and editing microscopy, which consists of the recording of taxonomically informative multifocal series of microscopy images as digital video clips. The integration of multifocal imaging with sequence analysis of the D2D3 region of large subunit (LSU) rDNA is illustrated here in the context of a combined morphological and barcode sequencing survey of marine nematodes from Baja California and California. The resulting video clips and sequence data are made available online in the database NemATOL (http://nematol.unh.edu/). Analyses of 37 barcoded nematodes suggest that these represent at least 32 species, none of which matches available D2D3 sequences in public databases. The recorded multifocal vouchers allowed us to identify most specimens to genus, and will be used to match specimens with subsequent species identifications and descriptions of preserved specimens. Like molecular barcodes, multifocal voucher archives are part of a wider effort at structuring and changing the process of biodiversity discovery. We argue that data-rich surveys and phylogenetic tools for analysis of barcode sequences are an essential component of the exploration of phyla with a high fraction of undiscovered species. Our methods are also directly applicable to other meiofauna such as for example gastrotrichs and tardigrades. PMID- 16214753 TI - Critical factors for assembling a high volume of DNA barcodes. AB - Large-scale DNA barcoding projects are now moving toward activation while the creation of a comprehensive barcode library for eukaryotes will ultimately require the acquisition of some 100 million barcodes. To satisfy this need, analytical facilities must adopt protocols that can support the rapid, cost effective assembly of barcodes. In this paper we discuss the prospects for establishing high volume DNA barcoding facilities by evaluating key steps in the analytical chain from specimens to barcodes. Alliances with members of the taxonomic community represent the most effective strategy for provisioning the analytical chain with specimens. The optimal protocols for DNA extraction and subsequent PCR amplification of the barcode region depend strongly on their condition, but production targets of 100K barcode records per year are now feasible for facilities working with compliant specimens. The analysis of museum collections is currently challenging, but PCR cocktails that combine polymerases with repair enzyme(s) promise future success. Barcode analysis is already a cost effective option for species identification in some situations and this will increasingly be the case as reference libraries are assembled and analytical protocols are simplified. PMID- 16214754 TI - A likelihood ratio test for species membership based on DNA sequence data. AB - DNA barcoding as an approach for species identification is rapidly increasing in popularity. However, it remains unclear which statistical procedures should accompany the technique to provide a measure of uncertainty. Here we describe a likelihood ratio test which can be used to test if a sampled sequence is a member of an a priori specified species. We investigate the performance of the test using coalescence simulations, as well as using the real data from butterflies and frogs representing two kinds of challenge for DNA barcoding: extremely low and extremely high levels of sequence variability. PMID- 16214757 TI - Current issues on infant nutrition. Proceedings of a symposium, Milano, Italy, 11 12 December 2003. PMID- 16214755 TI - TaxI: a software tool for DNA barcoding using distance methods. AB - DNA barcoding is a promising approach to the diagnosis of biological diversity in which DNA sequences serve as the primary key for information retrieval. Most existing software for evolutionary analysis of DNA sequences was designed for phylogenetic analyses and, hence, those algorithms do not offer appropriate solutions for the rapid, but precise analyses needed for DNA barcoding, and are also unable to process the often large comparative datasets. We developed a flexible software tool for DNA taxonomy, named TaxI. This program calculates sequence divergences between a query sequence (taxon to be barcoded) and each sequence of a dataset of reference sequences defined by the user. Because the analysis is based on separate pairwise alignments this software is also able to work with sequences characterized by multiple insertions and deletions that are difficult to align in large sequence sets (i.e. thousands of sequences) by multiple alignment algorithms because of computational restrictions. Here, we demonstrate the utility of this approach with two datasets of fish larvae and juveniles from Lake Constance and juvenile land snails under different models of sequence evolution. Sets of ribosomal 16S rRNA sequences, characterized by multiple indels, performed as good as or better than cox1 sequence sets in assigning sequences to species, demonstrating the suitability of rRNA genes for DNA barcoding. PMID- 16214758 TI - Fetal nutrition: a review. AB - Knowledge of fetal nutrient supply has greatly increased in the last decade due to the availability of fetal blood samples obtained under relatively steady-state conditions. These studies, together with studies utilizing stable isotope methodologies, have clarified some aspects of the supply of the major nutrients for the fetus such as glucose, amino acids and fatty acids. At the same time, the relevance of intrauterine growth has been recognized not only for the well-being of the neonate and child, but also for later health in adulthood. The major determinants of fetal nutrient availability are maternal nutrition and metabolism together with placental function and metabolism. The regulation of the rate of intrauterine growth is the result of complex interactions between genetic inheritance, endocrine environment and availability of nutrients to the fetus. PMID- 16214759 TI - Reproducing the bifidogenic effect of human milk in formula-fed infants: why and how? AB - Awareness of the key role of the intestinal microflora in the generation of the immunophysiological regulation and in the defence against pathogenic agents has attracted our interest in ways of manipulating the microbiota to improve health. Dietary modulation of the intestinal microflora is today one of the main topics of interest in the nutritional sciences. Performing this modulation in the neonatal or early infancy period, when immunological programming takes place, is a relatively new concept. Fructo-oligosaccharides (FOS) and galacto oligosaccharides (GOS) are prebiotics whose bifidogenic activity has been proven in adults. However, only recently have they been combined in infant formulas to reproduce the prebiotic effect of human milk oligosaccharides. In two consecutive trials, it has been demonstrated that supplementation of infant formulas with a mixture of GOS and FOS modified the fecal flora of term and preterm infants, stimulating the growth of Bifidobacteria. In the trial with term infants, the bifidogenic effect of the prebiotic mixture was dose dependent and there was also a significant increase in the number of Lactobacilli in the supplemented group. These findings offer a promising horizon for the early prevention of allergy and infections in infants. PMID- 16214760 TI - Prebiotic carbohydrates in human milk and formulas. AB - Human milk oligosaccharides play an important role, as prebiotic soluble fibres, in the postnatal development of the intestinal flora. Infant formulas are virtually free of prebiotic oligosaccharides. As a consequence, formula-fed infants develop an intestinal flora significantly different to the flora of breastfed infants. Due to the complexity of human milk oligosaccharides, it is necessary to use alternative sources of prebiotic ingredients as components of infant formulas. The present review summarizes the data of experimental research and clinical studies with a prebiotic mixture containing 90% short-chain galacto oligosaccharides and 10% long-chain fructo-oligosacchrides are summarized. The data demonstrate that, with this prebiotic mixture, the growth of bifidobacteria and lactobacilli can be stimulated, the faecal pH can be decreased, and the presence of pathogens can be reduced to levels similar to those of breastfed infants. Thus, prebiotic oligosaccharides such as the studied mixture provide beneficial effects for formula-fed infants. PMID- 16214762 TI - Dietary prebiotic oligosaccharides are detectable in the faeces of formula-fed infants. AB - Human milk oligosaccharides are not digested during intestinal passage and can be detected in stools. In this study it was investigated whether a prebiotic mixture of low-molecular-weight galacto-oligosaccharides (GOS) and high-molecular-weight fructo-oligosaccharides (FOS) can be detected in stool samples of formula-fed infants. The test formula was supplemented with 0.8 g/dl oligosaccharides (GOS+FOS). In the control formula, maltodextrins were used as placebo. Fecal flora was assessed at the beginning (day 1) and at the end of a 28-d feeding period (day 2). At day 2 the content of galacto- and fructo-oligosaccharides in the stool samples were measured. On study day 1, the number of bifidobacteria was not different among the groups (supplemented group: 7.7 (6.2) CFU/g; placebo group: 8.0 (6.0) CFU/g). At the end of the 28-d feeding period, the number of bifidobacteria was significantly higher in the group fed the supplemented formula when compared to placebo (supplemented group: 9.8 (0.7) CFU/g stool; placebo group: 7.1 (4.7) CFU/g stool; p<0.001). In all infants fed the supplemented formula, GOS and FOS could be identified in the stool samples. That was not the case in infants fed the non-supplemented formula. CONCLUSION: The present data confirm the bifidogenicity of oligosaccharides and indicate that dietary galacto oligosaccharides and long chain fructo-oligosaccharides remain during the whole passage in the lumen of the gastrointestinal tract, similarly to human milk oligosaccharides. PMID- 16214761 TI - Galacto-oligosaccharides and long-chain fructo-oligosaccharides as prebiotics in infant formulas: a review. AB - The present review summarizes clinical and experimental data concerning the possible effects of a prebiotic mixture of short-chain galacto-oligosaccharides and long-chain fructo-oligosaccharides. The results from several studies, made up of over 400 preterm and term infants, clearly demonstrate that the prebiotic mixture under examination specifically stimulates the growth of bifidobacteria and lactobacilli and reduces the growth of pathogens. As a consequence of the changed intestinal flora by the dietary galacto-oligosaccharides and fructo oligosaccharides, the faecal pH values and the short-chain fatty acid pattern were similar to those found in breastfed infants. In addition, the stool consistency was the same as in breastfed infants. In vitro experiments have demonstrated that the specific short-chain fatty acid pattern, at a pH similar to that found in faecal samples of breastfed infants, reduces the growth of pathogens in a dose-dependent manner but does not influence the growth of bifidobacteria and lactobacilli. In an animal vaccination model, the prebiotic mixture improved the response to vaccination. In an allergy model (sensitization by ovalbumin), the allergic reaction was reduced by the prebiotic mixture. The data obtained from animal experiments are in agreement with preliminary data from clinical trials which indicate a reduced allergic response (reduced plasma IgE/IgG4 ratio) and reduced episodes of upper airway infection during the first year of life. CONCLUSION: Experimental evidence demonstrates that the prebiotic mixture employed in these studies modulates the intestinal flora and modulates the immune system as human milk does. There are sufficient experimental data to put forward the hypothesis that substances like the prebiotic mixture under study will substantially contribute to the improvement of the protective properties of infant formulas. PMID- 16214763 TI - Increase of faecal bifidobacteria due to dietary oligosaccharides induces a reduction of clinically relevant pathogen germs in the faeces of formula-fed preterm infants. AB - In a previous study on formula-fed preterm infants, we were able to demonstrate that dietary oligosaccharides (a mixture of 90% galacto-oligosaccharides and 10% fructo-oligosaccharides in a concentration of 1 g/dl) stimulate the growth of faecal bifidobacteria. In the present explorative analysis of this study, we focus on the effect of the dominance of bifidobacteria on the presence of clinically relevant pathogens (Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Pseudomonas aeruginosa, Enterobacter, Klebsiella, Proteus, Streptococcus group B, Clostridium difficile, Bacillus subtilis and Acinetobacter). CONCLUSION: The data demonstrate that stimulation of bifidobacteria by prebiotic oligosaccharides reduces the presence of clinically relevant pathogens in the faecal flora, indicating that prebiotic substances might have the capacity to protect against enteral infections. PMID- 16214764 TI - Effects of specific lactic acid bacteria on the intestinal permeability to macromolecules and the inflammatory condition. AB - Non-live probiotic bacteria and their fermentation products can be used in milk based formula intended for healthy infants. The effects of a milk formula fermented with Bifidobacterium breve and Streptococcus thermophilus and heated/dehydrated to inactivate the micro-organisms have been reported over the last few years to decrease the intestinal permeability to macromolecules in experimental animals in vivo and more recently to down-regulate inflammatory condition in vitro. Feeding guinea-pigs with such dehydrated fermented milk reinforced the intestinal barrier resistance to food proteins (HRP, beta lactoglobulin). In addition, the products secreted by bacteria were capable of inhibiting the lipopolysaccharide (LPS)-induced TNF-alpha secretion by human peripheral mononuclear blood cells. The active secretion products were resistant to digestive enzymes and their anti-inflammatory properties were preserved after transepithelial transport across the filter-grown intestinal epithelial cell line, especially in inflammatory conditions. The binding of LPS to monocytes as well as NFkappaB nuclear translocation leading to pro-inflammatory cytokine transcription were inhibited by bacteria-culture supernatants. CONCLUSION: B. breve and S. thermophilus used as non-live micro-organisms in fermented infant formula seem to induce a reduction in macromolecular absorption and release metabolites exerting an anti-TNF-alpha effect, which persists after intestinal transport. Thus, specific lactic acid bacteria and their metabolites seem to affect positively the intestinal function. PMID- 16214765 TI - Nutritional requirements of the very preterm infant. PMID- 16214766 TI - Intravenous nutrition of the very preterm neonate. PMID- 16214767 TI - Considerations and approaches in determining the protein and energy composition of preterm infant formulas. AB - Optimal early nutritional support is considered a crucial issue in the care of the preterm infant, particularly of those with very low- or extremely low brithweight. Unfortunately, this goal is seldom satisfactorily attained. Several conditions such as hypoxia, acidosis, patent ductus arteriosus, drug therapy, reduced intestinal motility may interfere with an adequate nutritional delivery in the early neonatal period. Moreover, there is still concern about metabolic and intestinal tolerance of the currently suggested intakes and a lack of uniformity in the nutritional program among different NICUs. Finally, the vast majority of the available preterm formulas are not fully adequate to the real nutritional needs of these infants. Inadequate protein content and inappropriate protein energy ratio of most preterm formulas represent a matter of major concern, since there is a strict relationship between formula composition and the quantity and quality of weight gain. As a consequence there is a need of at least two milk formulas for different preterm infants: one for LBW and VLBW infants, the other for preterm infants weighing >1500 g. PMID- 16214768 TI - Human milk supplementation for preterm infants. AB - Nutrition support of the premature infant must be designed to compensate for metabolic and gastrointestinal immaturity, immunologic compromise, and associated medical conditions. The beneficial effects of human milk extend to the feeding of premature infants. However, nutritional concerns arise because the quantity of nutrients in human milk may not meet the great nutrient needs of the premature infant born weighing less than 1500 g. Human milk fortifiers are available to provide optimum nutrition. This review summarizes the benefits and limitations of human milk for the premature infant. PMID- 16214769 TI - Post-discharge nutrition for the preterm infant. AB - Newly designed, 'enriched,' formulas are now available for the formula feeding premature infant after hospital discharge. These new formulations add to the array of milks from which to choose when the discharge of the premature infant is approaching. However, the data to support optimal or reference nutrient intakes to use for the premature infant in the post-discharge are lacking. Breastfeeding should be encouraged as its beneficial effects may be related to the duration in which it is fed. Infants at risk for growth failure in the post-discharge period, such as infants unable to consume ad libitum intakes of milk, and who already have growth faltering or abnormal nutritional indices, may benefit for the addition of enriched formula. This review summarizes the recent data and comments on recommendations on post-discharge nutrition for premature infants. PMID- 16214770 TI - The effect of protective nutrients on mucosal defense in the immature intestine. AB - Oral nutrition plays a dual role in the gut, providing nutrition to the body while affecting the function of the gastrointestinal tract. The exposure of the gut to food antigens, in the form of either beneficial or harmful nutritional substances, contributes to a vast array of physiological and pathologic gastrointestinal responses with secondary systemic implications. The immune system of the gastrointestinal tract is always involved in the first line of defense, and its actions are particularly important in the early period of life as maturation takes place. From maturation, a balance ensues in the regulatory mechanism of host defense, ultimately leading to either tolerance or immune reaction. This paper emphasizes how some nutrients may beneficially affect the gastrointestinal immune system's maturation in both term and especially premature neonates. PMID- 16214771 TI - Hydrolysed milk in preterm infants: an open problem. AB - AIM: To review knowledge about the role, indications and nutritional adequacy of protein hydrolysate formulas (HF) in the preterm newborn. METHODS: A review of the available literature was carried out. RESULTS: No data support the use of HF for atopy prevention. HF could have a positive role in the early feeding of very low-birthweight infants, but several concerns exist about their nutritional adequacy. CONCLUSION: Further investigations addressing protein and mineral metabolism as well as long-term effects, including neurodevelopment, are needed. PMID- 16214772 TI - Calcium and phosphorus nutrition in preterm infants. AB - Metabolic bone disease is a common event in preterm infants. Between 24 wk gestation and term, the fetus accrues approximately 80% of body Ca, P and Mg. Consequently, preterm infants miss in part or completely the period of greatest mineral accretion. Ca and P requirements in preterm infants are based on demands for matching intrauterine bone mineral accretion rates and on maintenance of normal serum Ca and P concentrations. Daily physical therapy may be a useful adjunct, as it is associated with a significant increase in bone mineral density and content. Osteopenia/rickets of prematurity seems to be a self-resolving disease. However, this does not imply that a period of demineralization is acceptable. While the potential long-term consequences on peak bone mass are unclear at the moment, the short-term benefits include the avoidance of fractures, less marked dolicocephaly and improved linear growth. PMID- 16214773 TI - Feeding intolerance in very-low-birthweight infants: what is it and what can we do about it? AB - The increased survival of very-low-birth-weight infants has resulted in the need to better understand the immaturities that challenge optimal nutrition for these infants and how to surmount them. This is critical if we are to prevent short and long term morbidity associated with poor nutrition. Here we describe several of these immaturities including those related to digestion and absorption, suck swallow incoordination, delayed gastric emptying, and intestinal motility and how they lead to the common problem of feeding intolerance. Scientifically based strategies for introducing, stopping and continuing enteral feedings in association with parenteral nutrition are presented. PMID- 16214774 TI - Neonatal necrotizing enterocolitis: an update. AB - Necrotizing enterocolitis (NEC) is a leading cause of mortality and morbidity in neonatal intensive care units. Here we review selected manifestations of NEC, risk factors involved in its pathophysiology as well as putative mechanisms associated with how an immature gut might be more susceptible to NEC. Treatment and potential preventive strategies are discussed. PMID- 16214775 TI - What is the role of the prevention of allergy in the first period of life? AB - The widespread increase in the prevalence of allergic disease, which has occurred over the last 20 y, has created general concern in Europe and in the rest of the world. The reason for this increase is still partially unknown. In this period, despite a greater understanding of the pathogenesis and risk factors of allergies plus the greater efficacy of drugs to control the symptoms, the cost of intervention and the socio-economic impact are still very high. For these reasons, prevention in the first period of life represents a goal for both developed and developing countries in order to reduce this upward trend. PMID- 16214776 TI - Novel approaches to the nutritional management of the allergic infant. AB - The increased prevalence of atopic diseases, i.e. atopic eczema, allergic rhinitis and asthma, has been described as the epidemic of the 21st century in Western societies. New approaches in the fight against allergic diseases are clearly called for, the target being the persistence of the allergic responder pattern beyond infancy. The advantage afforded by elimination diets lies in the silencing of specific allergic inflammation induced by an offending food. Novel nutritional approaches, beyond the treatment of food allergies, have recently attracted research interest subsequent to the identification of the immunomodulatory potential of specific dietary compounds. Dietary lipids as immunomodulators may prevent allergic sensitization by down-regulating inflammatory response whilst protecting the epithelial barrier. Probiotic bacteria have been shown to reinforce the different lines of gut defence: immune exclusion, immune elimination and immune regulation. On this basis, the strategy against allergic disease proposed here is based on the administration of tolerogenic gut-processed peptide fragments of a specific protein, in addition to the use of specific dietary compounds such as fatty acids and antioxidants, and introducing a microbial stimulus for the immature immune system by means of cultures of beneficial live micro-organisms characteristic of the healthy infant gut microbiota. PMID- 16214777 TI - Z-score of weight for age of infants with atopic dermatitis and cow's milk allergy fed with a rice-hydrolysate formula during the first two years of life. AB - BACKGROUND: Recently, rice-based formulas have been widely used in hypoallergenic diets, but data on nutritional values are scarce. AIM: To evaluate the growth of infants fed with a rice-based hydrolysate formula, compared to those infants fed with a soy formula or an extensively hydrolysed casein formula, in the first 2 y of life. METHODS: A total of 88 infants were enrolled between March 2002 and March 2004. Fifty-eight infants with atopic dermatitis (AD) and cow's milk allergy (CMA), confirmed by open challenge, were enrolled as study group: 15 were fed with a rice-based hydrolysate formula (RHF), 17 with a soy-based formula (SF) and 26 with an extensively hydrolysed casein formula (eHCF). Thirty infants with AD without cow's milk allergy were recruited as a control group (CG) and fed with a free diet. Weight was recorded on enrolment and at 3-monthly intervals in the first year of life, and at 6-monthly intervals in the second year. Infants were weighed naked, before feeding, by means of an electronic integrating scale. The z scores of weight for age were calculated. STATISTICS: One-way analysis of variance and Student's t-test were used for statistical comparison. Significance was set at p<0.05. RESULTS: No significant differences between the RHF, SF and eHCF groups were observed for the z-score of weight for age during the first 2 y of life, but a significantly lower difference was seen in the RHF group compared to the control group in the intervals 9 mo-1 y (p=0.025) and 1-1.5 y (p=0.020) of age. In contrast, the SF and eHCF groups were comparable to the control group, but the eHCF group was significantly lower (p=0) in the first trimester of life. CONCLUSION: Even if our findings show no significant difference between RHF and control, low weight observed in infants fed with RHF raises doubts about the nutritional adequacy of rice-hydrolysate formulas. PMID- 16214778 TI - Advances in the management of digestive problems during the first months of life. AB - A study was carried out on 168 full-term infants with digestive problems such as regurgitation and/or constipation to evaluate the efficacy of new infant formulas containing partially hydrolysed whey protein, modified vegetable oil with a high beta-palmitic acid content, prebiotic oligosaccharides and starch. Infants receiving the new formulas had an increase in stool frequency of 0.60 between day 1 and day 7 (95% CI 0.19-1.01; p=0.004) and 0.53 (95% CI 0.11-0.90; p=0.015) between day 7 and day 14. A reduction of 1.06 in the number of regurgitation episodes was reported between day 1 and day 7 (95% CI 0.24-1.88; p=0.012) and 1.31 (95% CI 0.42-2.21; p=0.005) between day 7 and day 14. CONCLUSION: A prebiotic mixture of galacto-/fructo-oligosaccharides with a high beta-palmitic acid content may reduce digestive problems and improve intestinal tolerance in infants during the first months of life. PMID- 16214779 TI - Vitamin K prophylaxis: leaving the old route for the new one? AB - Oral or parenteral administration of vitamin K is the accepted practice for prevention of early vitamin K deficiency bleeding (VKDB) in the newborn. However, vitamin K prophylaxis in the newborn continues to be a worldwide health concern, particularly in breastfed infants. This paper reviews the current status of the use of vitamin K for the prevention of early and late VKDB. PMID- 16214780 TI - A prospective 10-year study on children who had severe infantile colic. AB - AIM: To evaluate the association between infantile colic and gastrointestinal, allergic and psychological disorders in childhood. METHODS: A prospective study was conducted on 103 infants aged 31-87 d. After 10 y, between 2001 and 2003, the children were recalled and a paediatrician evaluated the selected disorders by anamnesis, medical examination, laboratory tests and parent interviews. RESULTS: Of the 103 infants enrolled, 96 completed the study. There was an association between infantile colic and recurrent abdominal pain (p=0.001) and allergic disorders: allergic rhinitis, conjunctivitis, asthmatic bronchitis, pollenosis, atopic eczema and food allergy (p<0.05). Sleep disorders, fussiness, aggressiveness and feelings of supremacy are more frequent in children who suffered from colic during early infancy (p<0.05). A family history of gastrointestinal diseases and atopic diseases was significantly higher in infants with colic than in controls (p<0.05). CONCLUSION: Susceptibility to recurrent abdominal pain, allergic and psychological disorders in childhood may be increased by infantile colic. Our findings confirm that severe infantile colic might be the early expression of some of the most common disorders in childhood. PMID- 16214781 TI - Health research for development: making health research work ... for everyone. PMID- 16214782 TI - Dissolution stability studies of suspensions of prolonged-release diclofenac microcapsules prepared by the Wurster process: I. Eudragit-based formulation and possible drug-excipient interaction. AB - The aim was to evaluate possible interaction in solid and liquid state of the drug with formulation excipients consequent to very fast drug release of diclofenac-Eudragit prolonged release microcapsules. The microcapsules were prepared by drug layering on calcium carbonate cores and coated with Eudragit RS 30D and L30D-55 as previously reported. Suspension of the microcapsules was prepared using microcrystalline cellulose/sodium carboxymethyl cellulose (Avicel CL-611) as medium. In vitro dissolution testing of the suspension was done, and, based on the dissolution results, possible interaction between diclofenac and Eudragit and Avicel in the medium was studied. Powder X-ray diffraction (PXRD) and differential scanning calorimetry (DSC) analyses were performed using 1:1 binary, 1:1:1 ternary mixtures and a ratio equivalent to that in the formulation. The mixtures were prepared by mixing the dispersions--Eudragit RS 30D or L30D-55 with the drug or other components, followed by drying at 60 degrees C for 48 h. Dry mixing was done using the powder equivalents of the polymers, Eudragit RS PO and L100-55, Avicel and calcium carbonate. In vitro dissolution of the suspended microcapsules showed a very fast release after 48 h (T50 = <1 h) compared to the solid microcapsules (T50 = 6 h). DSC curves of the formulation components or microcapsules did not show the characteristic endothermic peak of diclofenac at 287 degrees C. Powder X-ray diffraction of the binary or ternary mixtures of diclofenac and Eudragit polymers indicated reduction, shift or modification of the crystalline peaks of the drug or excipients at 2theta of 12 degrees and 18 degrees , suggestive of interaction. Some changes in drug peak characteristics at 18 degrees and 23 degrees were observed for Avicel/drug mixture, though not significant. The DSC curves of the binary mixture of diclofenac co-dried with liquid forms of Eudragit (i.e. RS 30D or L30D-55) revealed greater interaction compared to the curves of drug and powdered forms of Eudragit (RS PO or L100-55). This was depicted by greater shift in fusion points of the mixtures relative to the drug. However, comparing the RS and L-type Eudragit, the latter generally showed greater interaction with the drug. Interaction between diclofenac and L type Eudragit polymers can occur in liquid formulations. PMID- 16214783 TI - Alginate-pectin microcapsules as a potential for folic acid delivery in foods. AB - Most naturally occurring folate derivatives in foods are highly sensitive to temperature, oxygen, light and their stability is affected by processing conditions. Folic acid incorporated microcapsules using alginate and combinations of alginate and pectin polymers were prepared to improve stability. Folic acid stability was evaluated with reference to encapsulation efficiency, gelling and hardening of capsules, capsular retention during drying and storage. Use of alginate in combination with pectin produced more robust capsules and contributed to greater encapsulation efficiency. The capsules lost their spherical shape as a consequence of increased pectin. The high alginate capsules, A100:P0 (100% alginate: 0% pectin) and A80:P20 (80% alginate: 20% pectin) were of regular spherical shape, while those with more pectin, A70:P30 (70% alginate: 30% pectin) and A60:P40 (60% alginate: 40% pectin) formed irregular spheres. The loading efficiency, expressed as a percentage of the actual loading to theoretical loading, varied from 55-89% with the composition of the mixed polymers. After 11 weeks of storage at 4 degrees C, folic acid retention in freeze-dried capsules was 100% (A70:P30 and A60:P40), 80% (A80:P20) and 30% (A100:P0). The blended alginate and pectin polymer matrix increased folic acid encapsulation efficiency and reduced the leakage from the capsules compared to those made with alginate alone and showed higher folic acid retention after freeze drying and storage. PMID- 16214784 TI - Suspensions of prolonged-release diclofenac-Eudragit and ion-exchange resin microcapsules: II. Improved dissolution stability. AB - PURPOSE: The stability of prolonged release 100 microm -size ion-exchange resin (IER) diclofenac microcapsules (prepared by the Wurster process) and coated with Eudragit RS30D was evaluated using dissolution analysis. METHODS: The IER microcapsules were suspended in 0.1% methylcellulose and stored at 23 and 37 degrees C and the dissolution study conducted over a 6-month period. The surface morphology of the microcapsules was examined using scanning electron microscopy (SEM). RESULTS: The dissolution of the suspensions stored at 23 degrees C on day 1 or 7 and was similar to that of day 30 with slightly faster dissolution on day 60. In contrast, release from suspensions stored at 37 degrees C decreased with storage. The decrease in dissolution with increased temperature was possibly due to the polymer relaxation (micromelting) that was enough to seal the drug within the matrix, resulting in slow dissolution. SEM of the suspended microcapsules correlated with the dissolution data, i.e. the surfaces of microcapsule stored at 37 degrees C showed decreased roughness or smoothening and closing of pores with time and, hence, retardation of drug release, compared with samples stored at 23 degrees C. The dissolution kinetics (shown by the linearity of Bt vs. time profiles) indicated that release mechanism was diffusion. CONCLUSIONS: The suspensions of diclofenac IER microcapsules were stable up to 30 days at ambient temperature, which makes the formulation potentially useful as reconstitutable product. PMID- 16214785 TI - Characteristics of vitamin C immobilized particles and sodium alginate beads containing immobilized particles. AB - This paper reports the properties of vitamin C encapsulated sodium alginate beads prepared by an alternative approach. The alternative encapsulation process mainly involves immobilization of vitamin C in hydrated zinc oxide layers and encapsulation of prepared immobilized particles in sodium alginate bead. The immobilization of vitamin C in hydrated zinc oxide layers was achieved by a coprecipitation process. Fourier transform infrared (FTIR) spectroscopy showed that the vitamin C was found to be stable after its immobilization. X-ray diffraction (XRD) studies revealed that anionic vitamin C molecules are adsorbed between positively charged zinc hydroxide layers with a 1:1 layer sequence, since well-defined change in basal spacing was observed. Well-defined change in surface morphology was observed by scanning electron microscopy (SEM) when vitamin C immobilized particles are encapsulated in sodium alginate bead. The biological activity of vitamin C was retained, even after its immobilization which was confirmed by 4-dihydroxy-L-phenylalanine (L-DOPA) oxidase inhibition and free radical scavenging activity studies. The release rate of vitamin C from immobilized particles and beads was sustained through an ion exchange process. A higher amount of stable vitamin C was recovered from the bead when compared to neat vitamin C itself. PMID- 16214786 TI - Preparation of cross-linked chitosan microspheres by spray drying: effect of cross-linking agent on the properties of spray dried microspheres. AB - Chitosan microspheres cross-linked with three different cross-linking agents viz, tripolyphosphate (TPP), formaldehyde (FA) and gluteraldehyde (GA) have been prepared by spray drying technique. The influence of these cross-linking agents on the properties of spray dried chitosan microspheres was extensively investigated. The particle size and encapsulation efficiencies of thus prepared chitosan microspheres ranged mainly between 4.1-4.7 microm and 95.12-99.17%, respectively. Surface morphology, % erosion, % water uptake and drug release properties of the spray dried chitosan microspheres was remarkably influenced by the type (chemical or ionic) and extent (1 or 2% w/w) of cross-linking agents. Spray dried chitosan microspheres cross-linked with TPP exhibited higher swelling capacity, % water uptake, % erosion and drug release rate at both the cross linking extent (1 and 2% w/w) when compared to those cross-linked with FA and GA. The sphericity and surface smoothness of the spray dried chitosan microspheres was lost when the cross-linking extent was increased from 1 to 2% w/w. Release rate of the drug from spray dried chitosan microspheres decreased when the cross linking extent was increased from 1 to 2% w/w. The physical state of the drug in chitosan-TPP, chitosan-FA and chitosan-GA matrices was confirmed by the X-ray diffraction (XRD) study and found that the drug remains in a crystalline state even after its encapsulation. Release of the drug from chitosan-TPP, chitosan-FA and chitosan-GA matrices followed Fick's law of diffusion. PMID- 16214787 TI - Application of self-assembled ultra-thin film coatings to stabilize macromolecule encapsulation in alginate microspheres. AB - Alginate-based hydrogels have several unique properties that have enabled them to be used as a matrix for the entrapment of a variety of enzymes, proteins and cells for applications in bioprocessing, drug delivery and chemical sensing. However, control over release rates or, in some cases, stable encapsulation remains a difficult goal, especially for small particles with high surface-area to-volume ratios. In this work, the potential to limit diffusion of macromolecules embedded in alginate spheres with nanofilm coatings was assessed. Alginate microspheres were fabricated using an emulsification process with high surfactant concentration to form beads in the size range of 2-10 microm. Using calcium chloride for ionotropic gelation, dextran was encapsulated in the gel phase by mixing with the alginate in solution. The exterior surface was then modified with polyelectrolyte coatings using the layer-by-layer self assembly technique. Leaching studies to assess retention of dextran with varying molecular weights confirmed that the application of multi-layer thin films to the alginate microspheres was effective in reducing leaching rate and total loss of the encapsulated material from the microspheres. For the best case, the rate of release for dextran of 2,000,000 Dalton molecular weight decreased from 1% h(-1) in bare microspheres to 0.1% h(-1) in polyelectrolyte-coated microspheres. The effectiveness of nanofilms reducing loss of the encapsulated macromolecules was found to vary between different polycation materials used. These studies support the feasibility of using these microsystems for development of long-term stable encapsulated systems, such as implantable biosensors. PMID- 16214788 TI - Optimizing double emulsion process to decrease the burst release of protein from biodegradable polymer microspheres. AB - The process parameters such as the compositions of inner and outer aqueous phase and emulsification technique of the primary emulsion were optimized to decrease the burst release of BSA from biodegradable polymer microspheres in double emulsion method. It was found that diminished burst release of -14% was achieved for the microspheres produced by formulations, where no phosphate was present in the inner water phase (non-buffered system). Primary emulsion made by probe sonication rather than homogenization or mechanical stirring led to microspheres with insignificant burst effect. Microspheres obtained using 0.1% aqueous Tween 80 solution as the outer aqueous phase, frequently exhibit reduced burst effect of 2.7%. Low microsphere yield (52.1%), however, was observed. Microsphere yield was, therefore, enhanced by addition of additive such as sodium chloride, glucose or mannitol into the outer aqueous phase. Decrease in BSA entrapment was observed in the presence of sodium chloride, but reduction in entrapment efficiency was observed in the case of glucose. Burst release increased from 2.7% to 9.5% or 3.4% as 2.5% sodium chloride or 7.5% glucose was added into the outer aqueous phase respectively. Marked burst release (>20%) was observed in the presence of additive of higher concentration independent of sodium chloride or glucose. As far as surfactant type was concerned, diminished burst was found when PVP or Tween 80 rather than PVA was utilized as the surfactant during microsphere preparation. In addition to PLGA, the copolymers of L-lactide (LLA) and dimethyl trimethylene carbonate (DTC) or trimethylene carbonate (TMC) were also evaluated. Insignificant burst effect was found for the microspheres composed of DTC or TMC copolymers. PMID- 16214791 TI - Literature alerts. PMID- 16214789 TI - Use of porous aluminosilicate pellets for drug delivery. AB - Three pelletized porous aluminosilicate ceramics were obtained commercially and their potential to act as extended release drug delivery systems was assessed. The pellets were drug loaded using a vacuum impregnation technique. Factors such as the concentration of the loading solution and the porosity and bulk density of the ceramic influenced the drug loading. The release of drug from the pellets was extended as the drug was entrapped within their porous interior. The rate of release was influenced by the porous microstructure of the pellets and the physicochemical properties of the drug. Extrusion-spheronization was used to prepare pellets similar to the porous ceramics. The pellet formulations contained an aluminosilicate clay mineral (kaolin or halloysite), ethylcellulose 100 cps, ethanol and varying quantities of sucrose. The latter two components acted as pore forming agents. Diltiazem HCl was loaded into the pellets and its release was extended. The release rate could be modified by changing the quantity of sucrose included in the initial formulation, as this influenced the porous microstructure of the pellets. In halloysite-based products the release was further extended due to entrapment of the drug within the halloysite microtubules. Porous kaolin-based pellets were also prepared by cryopelletization. This involved freezing droplets of an aqueous suspension containing kaolin, sodium silicate solution and sodium lauryl sulphate. The resulting pellets were freeze-dried, which removed ice from them to leave pores behind. The pellets gave extended drug release with the release rate being influenced by the porous microstructure of the pellets and their microclimate pH. PMID- 16214793 TI - "Doing it properly": the experience of first mothering over 35 years. AB - First mothering over 35 years is an increasing phenomenon in developed countries, and this "greying" of maternity raises some interesting questions and dilemmas for clinical care. In a qualitative study conducted in Melbourne, Australia, the motherhood experiences of 22 primiparous women were explored. Participant age ranged from 35 to 48 years. Women were interviewed over three junctures: at 35-38 weeks gestation, 10-14 days postpartum, and 8 months postpartum. Becoming a mother was found to occur in a temporally ordered sequence, with clear markers at 1-4 weeks, 1-4 months, 4-6 months, and 6-8 months. Themes that emerged from the analysis included the project "doing it properly," vulnerability, "finding my own way," and "being older." The continuing and increasing trend of primiparity older than 35 years makes this account of interest globally. Findings from this study offer an alternative explanation of early mothering over 35 years and offers direction to health professionals for easing early motherhood experiences for this growing group of childbearing women. PMID- 16214794 TI - Meanings of being pregnant and having decided on abortion: young Swedish women's experiences. AB - In-depth interviews about the experience of being pregnant and having decided on abortion were conducted with 10 young Swedish women. A phenomenological hermeneutic study disclosed a complex life world summarised as "I'm happy about my fertility but I choose not to give life now." Four themes were revealed: encountering an unexpected understanding of oneself and one's fertility, desiring to be understood by trusted others, trying out different positions in relation to pregnancy and abortion, and approaching planned adult motherhood. The results indicate that addressing young women's concerns about fertility might be important in reproductive care. PMID- 16214795 TI - Identifying healthy eating strategies in low-income pregnant women: applying a positive deviance model. AB - Using positive deviance methodology, we identified strategies that enabled some low-income pregnant women to eat healthy meals while others did not. We used a descriptive design consisting of small group interviews of low-income pregnant women and identified 6 of 18 women as eating healthy diets. Women with healthy diets knew to eat balanced meals, had family support, were willing to prepare foods that were different than other family members, and ate at home more frequently than women with unhealthy diets. Health care providers can use the positive deviance approach to guide the development of interventions to improve women's diets using community-specific solutions to enhance the health of mothers and infants. PMID- 16214797 TI - Is older maternal age a risk factor for preterm birth and fetal growth restriction? A systematic review. AB - To determine if there was an association between advancing maternal age and adverse pregnancy outcomes (preterm delivery and small-for-gestational-age births), a systematic review was conducted based on a comprehensive search of the literature from 1985 to 2002. Ten studies met the following inclusion criteria: (1) assessed risk factors for preterm birth by subtype (i.e., idiopathic preterm labor, preterm premature rupture of membranes) and small-for-gestational-age (SGA) birth (fetal growth restriction); (2) used acceptable definitions of these outcomes; (3) were published between January 1985 and December 2002; (4) were restricted to studies that have considered preterm birth due to idiopathic preterm labor or premature rupture of membranes or both; (5) were restricted to singleton live births; (6) were conducted in a developed country; and (7) were published in English. The majority of the studies reviewed found that older maternal age was associated with preterm birth. There is insufficient evidence to determine if older maternal age is an independent and direct risk factor for preterm birth and SGA birth, or a risk marker that exerts its influence on gestational age or birth weight or both through its association with age dependent confounders. Future research is needed to quantify the independent and unconfounded impact of delayed childbearing on neonatal outcomes, as well as to identify the pathways involved. PMID- 16214796 TI - Having a "safe delivery": conflicting views from Tibet. AB - In the Tibetan Autonomous Region (TAR) of the People's Republic of China (PRC) maternal mortality ratios remain among the highest in the world. Although traditional Tibetan medical theory, practice, and pharmacology include information on maternal and child health care, Tibet is one of the few societies in the world that does not have traditional birth attendants or midwives. Using ethnographic methods, we gathered data from individual interviews with rural Tibetan women (N=38) about their beliefs and behaviors surrounding pregnancy and childbirth. Additional data were gathered through interviews with prefecture, county, and township health care providers. These data were used to develop a culturally appropriate village birth attendant training program in rural Tibet. We describe Tibetan women's perspectives of "having a safe delivery" in relation to concepts about "safe delivery" according to evidence-based medicine in the West. Our work also provides an example of the benefits and challenges that arise when ethnographic research methods are used to design and implement health care interventions. PMID- 16214800 TI - PCR amplification from single DNA molecules on magnetic beads in emulsion: application for high-throughput screening of transcription factor targets. AB - We have developed a novel method of genetic library construction on magnetic microbeads based on solid-phase single-molecule PCR in a fine and robust water phase compartment formed in water-in-oil (w/o) emulsions. In this method, critically diluted DNA fragments were distributed over the emulsion as templates, where beads crosslinked with multiple primers and other PCR components were encapsulated to form multiple reaction compartments. The delivered DNA was then amplified and covalently immobilized on the beads in parallel, within individual compartments, to construct a genetic library on beads (GLOBE), which was readily applicable to a genomewide global scanning of genetic elements recognized by a defined DNA-binding protein. We constructed a GLOBE of Paracoccus denitrificans and selected gene beads that were bound to the His-tagged transcription factor PhaR by flow cytometry. As a result of flow cytometry screening with an anti-His fluorescent antibody, the PhaR target fragments were enriched 1200-fold from this library with this system. Therefore, this system is a powerful tool for analyzing the transcription network on a genomewide scale. PMID- 16214801 TI - Nucleic acid melting by Escherichia coli CspE. AB - Escherichia coli contains nine members of the CspA family. CspA and some of its homologues play critical role in cold acclimation of cells by acting as RNA chaperones, destabilizing nucleicacid secondary structures. Disruption of nucleic acid melting activity of CspE led to loss of its transcription antitermination activity and consequently its cold acclimation activity. To date, the melting activity of Csp proteins was studied using partially double-stranded model nucleic acids substrates forming stem-loop structures. Here, we studied the mechanism of nucleic acid melting by CspE. We show that CspE melts the stem region in two directions, that CspE-induced melting does not require the continuity of the substrate's loop region, and CspE can efficiently melt model substrates with single-stranded overhangs as short as 4 nt. We further show that preferential binding of CspE at the stem-loop junction site initiates melting; binding of additional CspE molecules that fully cover the single-stranded region of a melting substrate leads to complete melting of the stem. PMID- 16214803 TI - The subsystems approach to genome annotation and its use in the project to annotate 1000 genomes. AB - The release of the 1000th complete microbial genome will occur in the next two to three years. In anticipation of this milestone, the Fellowship for Interpretation of Genomes (FIG) launched the Project to Annotate 1000 Genomes. The project is built around the principle that the key to improved accuracy in high-throughput annotation technology is to have experts annotate single subsystems over the complete collection of genomes, rather than having an annotation expert attempt to annotate all of the genes in a single genome. Using the subsystems approach, all of the genes implementing the subsystem are analyzed by an expert in that subsystem. An annotation environment was created where populated subsystems are curated and projected to new genomes. A portable notion of a populated subsystem was defined, and tools developed for exchanging and curating these objects. Tools were also developed to resolve conflicts between populated subsystems. The SEED is the first annotation environment that supports this model of annotation. Here, we describe the subsystem approach, and offer the first release of our growing library of populated subsystems. The initial release of data includes 180 177 distinct proteins with 2133 distinct functional roles. This data comes from 173 subsystems and 383 different organisms. PMID- 16214802 TI - Crystal structures of complexes between aminoglycosides and decoding A site oligonucleotides: role of the number of rings and positive charges in the specific binding leading to miscoding. AB - The crystal structures of six complexes between aminoglycoside antibiotics (neamine, gentamicin C1A, kanamycin A, ribostamycin, lividomycin A and neomycin B) and oligonucleotides containing the decoding A site of bacterial ribosomes are reported at resolutions between 2.2 and 3.0 A. Although the number of contacts between the RNA and the aminoglycosides varies between 20 and 31, up to eight direct hydrogen bonds between rings I and II of the neamine moiety are conserved in the observed complexes. The puckered sugar ring I is inserted into the A site helix by stacking against G1491 and forms a pseudo base pair with two H-bonds to the Watson-Crick sites of the universally conserved A1408. This central interaction helps to maintain A1492 and A1493 in a bulged-out conformation. All these structures of the minimal A site RNA complexed to various aminoglycosides display crystal packings with intermolecular contacts between the bulging A1492 and A1493 and the shallow/minor groove of Watson-Crick pairs in a neighbouring helix. In one crystal, one empty A site is observed. In two crystals, two aminoglycosides are bound to the same A site with one bound specifically and the other bound in various ways in the deep/major groove at the edge of the A sites. PMID- 16214804 TI - A bi-functional siRNA construct induces RNA interference and also primes PCR amplification for its own quantification. AB - RNA interference (RNAi) is a process of post-transcriptional gene silencing initiated by double-stranded RNAs, including short interfering RNA (siRNA). Silencing is sequence-specific and RNAi has rapidly become central to the study of gene function. RNAi also carries promise for selective silencing of viral and endogenous genes causal for disease. To detect the very low levels of siRNA effective for RNAi we modified the 3' end of the sense strand of siRNA with a nuclease-resistant DNA hairpin. We show that the modified siRNA-DNA construct (termed 'crook' siRNA) functions as a primer for the PCR and describe a novel, yet simple PCR protocol for its quantification (amolar levels/cell). When transfected into mammalian cells, crook siRNA induces selective mRNA knock-down equivalent to its unmodified siRNA counterpart. This new bifunctional siRNA construct will enable future in vivo studies on the uptake, distribution and pharmacokinetics of siRNA, and is particularly important for the development of siRNA-based therapeutics. More generally, PCR-based detection of siRNA carries wide-ranging applications for RNAi reverse genetics. PMID- 16214805 TI - A highly sensitive selection method for directed evolution of homing endonucleases. AB - Homing endonucleases are enzymes that catalyze DNA sequence specific double strand breaks and can significantly stimulate homologous recombination at these breaks. These enzymes have great potential for applications such as gene correction in gene therapy or gene alteration in systems biology and metabolic engineering. However, homing endonucleases have a limited natural repertoire of target sequences, which severely hamper their applications. Here we report the development of a highly sensitive selection method for the directed evolution of homing endonucleases that couples enzymatic DNA cleavage with the survival of host cells. Using I-SceI as a model homing endonuclease, we have demonstrated that cells with wild-type I-SceI showed a high cell survival rate of 80-100% in the presence of the original I-SceI recognition site, whereas cells without I SceI showed a survival rate <0.003%. This system should also be readily applicable for directed evolution of other DNA cleavage enzymes. PMID- 16214806 TI - Improved methods for the generation of human gene knockout and knockin cell lines. AB - Recent studies have demonstrated the utility of recombinant adeno-associated viral (rAAV) vectors in the generation of human knockout cell lines. The efficiency with which such cell lines can be generated using rAAV, in comparison with more extensively described plasmid-based approaches, has not been directly tested. In this report, we demonstrate that targeting constructs delivered by rAAV vectors were nearly 25-fold more efficient than transfected plasmids that target the same exon. In addition, we describe a novel vector configuration which we term the synthetic exon promoter trap (SEPT). This targeting element further improved the efficiency of knockout generation and uniquely facilitated the generation of knockin alterations. An rAAV-based SEPT targeting construct was used to transfer a mutant CTNNB1 allele, encoding an oncogenic form of beta catenin, from one cell line to another. This versatile method was thus shown to facilitate the efficient integration of small, defined sequence alterations into the chromosomes of cultured human cells. PMID- 16214807 TI - Dominant-negative mutant phenotypes and the regulation of translation elongation factor 2 levels in yeast. AB - The eukaryotic translation elongation factor 2 (eEF2), a member of the G-protein superfamily, catalyzes the post-peptidyl transferase translocation of deacylated tRNA and peptidyl tRNA to the ribosomal E- and P-sites. eEF2 is modified by a unique post-translational modification: the conversion of His699 to diphthamide at the tip of domain IV, the region proposed to mimic the anticodon of tRNA. Structural models indicate a hinge is important for conformational changes in eEF2. Mutations of V488 in the hinge region and H699 in the tip of domain IV produce non-functional mutants that when co-expressed with the wild-type eEF2 result in a dominant-negative growth phenotype in the yeast Saccharomyces cerevisiae. This phenotype is linked to reduced levels of the wild-type protein, as total eEF2 levels are unchanged. Changes in the promoter, 5'-untranslated region (5'-UTR) or 3'-UTR of the EFT2 gene encoding eEF2 do not allow overexpression of the protein, showing that eEF2 levels are tightly regulated. The H699K mutant, however, also alters translation phenotypes. The observed regulation suggests that the cell needs an optimum amount of active eEF2 to grow properly. This provides information about a new mechanism by which translation is efficiently maintained. PMID- 16214808 TI - The physical determinants of the DNA conformational landscape: an analysis of the potential energy surface of single-strand dinucleotides in the conformational space of duplex DNA. AB - A multivariate analysis of the backbone and sugar torsion angles of dinucleotide fragments was used to construct a 3D principal conformational subspace (PCS) of DNA duplex crystal structures. The potential energy surface (PES) within the PCS was mapped for a single-strand dinucleotide model using an empirical energy function. The low energy regions of the surface encompass known DNA forms and also identify previously unclassified conformers. The physical determinants of the conformational landscape are found to be predominantly steric interactions within the dinucleotide backbone, with medium-dependent backbone-base electrostatic interactions serving to tune the relative stability of the different local energy minima. The fidelity of the PES to duplex DNA properties is validated through a correspondence to the conformational distribution of duplex DNA crystal structures and the reproduction of observed sequence specific propensities for the formation of A-form DNA. The utility of the PES is demonstrated through its succinct and accurate description of complex conformational processes in simulations of duplex DNA. The study suggests that stereochemical considerations of the nucleic acid backbone play a role in determining conformational preferences of DNA which is analogous to the role of local steric interactions in determining polypeptide secondary structure. PMID- 16214809 TI - Psychosocial effects of the 2001 UK foot and mouth disease epidemic in a rural population: qualitative diary based study. AB - OBJECTIVES: To understand the health and social consequences of the 2001 foot and mouth disease epidemic for a rural population. DESIGN: Longitudinal qualitative analysis. SETTING: North Cumbria, the worst affected area in Britain. SAMPLE: Purposive sample of 54 respondents divided in six demographically balanced rural occupational and population groups. MAIN OUTCOME MEASURES: 3071 weekly diaries contributed over 18 months; 72 semistructured interviews (with the 54 diarists and 18 others); 12 group discussions with diarists RESULTS: The disease epidemic was a human tragedy, not just an animal one. Respondents' reports showed that life after the foot and mouth disease epidemic was accompanied by distress, feelings of bereavement, fear of a new disaster, loss of trust in authority and systems of control, and the undermining of the value of local knowledge. Distress was experienced across diverse groups well beyond the farming community. Many of these effects continued to feature in the diaries throughout the 18 month period. CONCLUSIONS: The use of a rural citizens' panel allowed data capture from a wide spectrum of the rural population and showed that a greater number of workers and residents had traumatic experiences than has previously been reported. Recommendations for future disaster management include joint service reviews of what counts as a disaster, regular NHS and voluntary sector sharing of intelligence, debriefing and peer support for front line workers, increased community involvement in disposal site or disaster management, and wider, more flexible access to regeneration funding and rural health outreach work. PMID- 16214810 TI - Characterization of human phagocyte-derived chitotriosidase, a component of innate immunity. AB - Man has been found to produce highly conserved chitinases. The most prominent is the phagocyte-derived chitotriosidase, the plasma levels of which are markedly elevated in some pathological conditions. Here, we report that both polymorphonuclear neutrophils (PMNs) and macrophages (m) are a source of chitotriosidase. The enzyme is located in specific granules of human PMNs and secreted following stimulation with granulocyte macrophage colony-stimulating factor (GM-CSF). In addition, GM-CSF induces expression of chitotriosidase in m that constitutively secrete the enzyme and partly accumulate it in their lysosomes. Studies with recombinant human chitotriosidase revealed that the enzyme targets chitin-containing fungi. These findings are consistent with earlier observations concerning anti-fungal activity of homologous plant chitinases and beneficial effects of GM-CSF administration in individuals suffering from invasive fungal infections. In conclusion, chitotriosidase should be viewed as a component of the innate immunity that may play a role in defence against chitin-containing pathogens and the expression and release of which by human phagocytes is highly regulated. PMID- 16214811 TI - Regulation of the type I IFN induction: a current view. AB - The type I IFN-alpha/beta gene family was identified about a quarter of a century ago as a prototype of many cytokine gene families, which led to the subsequent burst of studies on molecular mechanisms underlying cytokine gene expression and signaling. Although originally discovered for their activity to confer an antiviral state on cells, more evidence has recently been emerging regarding IFN alpha/beta actions on cell growth, differentiation and many immunoregulatory activities, which are of even greater fundamental biological significance. Indeed, much attention has recently been focused on the induction and function of the IFN-alpha/beta system regulated by Toll-like receptors (TLRs), which are critical for linking the innate and adaptive immunities. The understanding of the regulatory mechanisms of IFN-alpha/beta gene induction by TLRs and viruses is an emerging theme, for which much new insight has been gained over the past few years. PMID- 16214812 TI - Tomato juice prevents senescence-accelerated mouse P1 strain from developing emphysema induced by chronic exposure to tobacco smoke. AB - The senescence-accelerated mouse (SAM) is a naturally occurring animal model for accelerated aging after normal development and maturation. SAMP1 strain was reported to show age-related structural and functional changes in lung and to be a murine model of senile lung. We postulated that aging of lung is an important intrinsic process for development of emphysema and even in a short period of tobacco smoke exposure may be able to generate emphysema. At age 12 wk, SAMP1 inhaled air or 1.5% tobacco smoke (total particulate matter 23.9 mg/m3) through the nose for 30 min/day, 5 days/wk, and for 8 wk. The mean linear intercepts (MLI) and destructive index (DI) of lung were significantly increased [air vs. smoke (means+/-SE); MLI, 68.76+/-0.69 vs. 75.34+/-1.70 microm, P<0.05 and DI, 8.61+/-0.38 vs. 16.18+/-1.54%, P<0.05], whereas no significant changes were observed in SAMR1, control mice that show normal aging. In contrast, smoke induced emphysema was completely prevented by concomitant ingestion of lycopene given as tomato juice [MLI: smoke with/without lycopene (mean+/-SE), 62.87+/-0.8 vs. 66.90+/-1.33 microm, P<0.05]. Smoke exposure increased apoptosis and active caspase-3 of airway and alveolar septal cells and reduced VEGF in lung tissues, but tomato juice ingestion significantly reduced apoptosis and increased tissue VEGF level. We conclude that SAMP1 is a useful model for tobacco smoke-induced emphysema and a valuable tool to explore both pathophysiological mechanisms and the effect of therapeutic intervention on smoke-induced emphysema. PMID- 16214813 TI - Differential degradation of matrix proteoglycans and edema development in rabbit lung. AB - The specific role of solid extracellular matrix components in opposing development of pulmonary interstitial edema was studied in adult anesthetized rabbits by challenging the lung parenchyma with an intravenous injection of a bolus of collagenase or heparanase. In 10 rabbits, pulmonary interstitial pressure (Pip) was measured by micropuncture in control and up to 3 h after collagenase or heparanase intravenous injection. With respect to control (Pip= 9.3 +/- 1.5 cmH2O, n = 10), both treatments caused a significant increase of Pip and of the wet weight-to-dry weight lung ratio. However, while tissue matrix stiffness was maintained after 60 min of collagenase, as indicated by the attainment of a positive Pip peak (Pip= 4.5 +/- 0.3 cmH2O, n = 5), this mechanical response was lost with heparanase (Pip= -0.6 +/- 1.3 cmH2O, n = 5). Biochemical analysis performed on a separate rabbit group (n = 15) showed an increased extraction of uronic acid with both enzymes, indicating a progressive matrix fragmentation. Gel chromatography analysis of the proteoglycan (PG) families showed that 60 min of both enzymatic treatments left the large-molecular weight PGs (versican) essentially unaffected. However, the heparan-sulfate PG fraction was significantly cleaved, as indicated by a significant increase of the smaller PG fragments with heparanase, but not with collagenase. Hence, present data suggest that the integrity of the heparan-sulfate PGs is required to maintain the three-dimensional architecture of the pulmonary tissue matrix and in turn to counteract tissue fluid accumulation in situations of increased fluid filtration. PMID- 16214814 TI - Hypoxia induces hypersensitivity and hyperreactivity to thromboxane receptor agonist in neonatal pulmonary arterial myocytes. AB - PPHN, caused by perinatal hypoxia or inflammation, is characterized by an increased thromboxane-prostacyclin ratio and pulmonary vasoconstriction. We examined effects of hypoxia on myocyte thromboxane responsiveness. Myocytes from 3rd-6th generation pulmonary arteries of newborn piglets were grown to confluence and synchronized in contractile phenotype by serum deprivation. On the final 3 days of culture, myocytes were exposed to 10% O2 for 3 days; control myocytes from normoxic piglets were cultured in 21% O2. PPHN was induced in newborn piglets by 3-day hypoxic exposure (Fi(O2) 0.10); pulmonary arterial myocytes from these animals were maintained in normoxia. Ca2+ mobilization to thromboxane mimetic U-46619 and ATP was quantified using fura-2 AM. Three-day hypoxic exposure in vitro results in increased basal [Ca2+]i, faster and heightened peak Ca2+ response, and decreased U-46619 EC50. These functional changes persist in myocytes exposed to hypoxia in vivo but cultured in 21% O2. Blockade of Ca2+ entry and store refilling do not alter peak U-46619 Ca2+ responses in hypoxic or normoxic myocytes. Blockade of ryanodine-sensitive or IP3-gated intracellular Ca2+ channels inhibits hypoxic augmentation of peak U-46619 response. Ca2+ response to ryanodine alone is undetectable; ATP-induced Ca2+ mobilization is unaltered by hypoxia, suggesting no independent increase in ryanodine-sensitive or IP3-linked intracellular Ca2+ pool mobilization. We conclude hypoxia has a priming effect on neonatal pulmonary arterial myocytes, resulting in increased resting Ca2+, thromboxane hypersensitivity, and hyperreactivity. We postulate that hypoxia increases agonist-induced TP-R-linked IP3 pathway activation. Myocyte thromboxane hyperresponsiveness persists in culture after removal from the initiating hypoxic stimulus, suggesting altered gene expression. PMID- 16214816 TI - Rat pulmonary arterial smooth muscle myosin light chain kinase and phosphatase activities decrease with age. AB - We and others have shown that the fetal pulmonary arterial smooth muscle potential for contraction and relaxation is significantly reduced compared with the adult. Whether these developmental changes relate to age differences in the expression and/or activity of key enzymes regulating the smooth muscle mechanical properties has not been previously evaluated. Therefore, we studied the catalytic activities and expression of myosin light chain kinase (MLCK) and myosin light chain phosphatase (MLCP) catalytic (PP1cdelta) and regulatory (MYPT) subunits in late fetal, early newborn, and adult rat intrapulmonary arterial tissues. In keeping with the greater force development and relaxation of adult pulmonary artery, Western blot analysis showed that the MLCK, MYPT, and PP1cdelta contents increased significantly with age and were highest in the adult rat. In contrast, their specific activities (activity/enzyme content) were significantly higher in the fetal compared with the adult tissue. The fetal and newborn pulmonary arterial muscle relaxant response to the Rho-kinase inhibitor Y-27632 was greater than the adult tissue. In addition to the 130-kDa isoform of MLCK, we documented the presence of minor higher-molecular-weight embryonic isoforms in the fetus and newborn. During fetal life, the lung pulmonary arterial MLCK- and MLCP-specific activities are highest and appear to be related to Rho-kinase activation during lung morphogenesis. PMID- 16214815 TI - Constitutive activation of prosurvival signaling in alveolar mesenchymal cells isolated from patients with nonresolving acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) is a clinical syndrome characterized by stereotypic host inflammatory and repair cellular responses; however, mechanisms regulating the resolution of ARDS are poorly understood. Here, we report the isolation and characterization of a novel population of mesenchymal cells from the alveolar space of ARDS patients via fiber-optic bronchoscopy with bronchoalveolar lavage (BAL). BAL was performed on 17 patients during the course of ARDS. Immunofluorescence staining and multiparameter flow cytometric analysis defined a population of alveolar mesenchymal cells (AMCs) that are CD45-/prolyl-4 hydroxylase+/alpha-smooth muscle actin+/-. AMCs proliferated in ex vivo cell culture for multiple passages; early passage (3-5) cells were subsequently analyzed in 13 patients. AMCs isolated from patients with persistent or nonresolving ARDS (ARDS-NR, n = 4) demonstrate enhanced constitutive activation of prosurvival signaling pathways involving PKB/Akt, FKHR, and BCL-2 family proteins compared with AMCs from patients with resolving ARDS (ARDS-R, n = 9). Exogenous transforming growth factor-beta1 markedly induces PKB/Akt activation in AMCs from ARDS-R. ARDS-NR cells are more resistant to serum deprivation-induced apoptosis compared with ARDS-R. This study identifies a novel population of mesenchymal cells that can be isolated from the alveolar spaces of ARDS patients. AMCs in patients with ARDS-NR acquire an activational profile characterized by enhanced prosurvival signaling and an antiapoptotic phenotype. These findings support the concept that apoptosis of mesenchymal cells may be an essential component of normal repair and resolution of ARDS and suggest that dysregulation of this process may contribute to persistent ARDS. PMID- 16214817 TI - Congenital diaphragmatic hernia prevents absorption of distal air space fluid in late-gestation rat fetuses. AB - We hypothesized that congenital diaphragmatic hernia (CDH) may decrease distal air space fluid absorption due to immaturity of alveolar epithelial cells from a loss of the normal epithelial Na+ transport, as assessed by amiloride and epithelial Na+ channel (ENaC) and Na-K-ATPase expression, as well as failure to respond to endogenous epinephrine as assessed by propranolol. Timed-pregnant dams were gavage fed 100 mg of nitrofen at 9.5-day gestation to induce CDH in the fetuses, and distal air space fluid absorption experiments were carried out on 22 day gestation (term) fetuses. Controls were nitrofen-exposed fetuses without CDH. Absorption of distal air space fluid was measured from the increase in 131I albumin concentration in an isosmolar, physiological solution instilled into the developing lungs. In controls, distal air space fluid absorption was rapid and mediated by beta-adrenoceptors as demonstrated by reversal to fluid secretion after propranolol. Normal lung fluid absorption was also partially inhibited by amiloride. In contrast, CDH fetuses continued to show lung fluid secretion, and this secretion was not affected by either propranolol or amiloride. CDH lungs showed a 67% reduction in alpha-ENaC and beta-ENaC expression, but no change in alpha1-Na-K-ATPase expression. These studies demonstrate: 1) CDH delays lung maturation with impaired distal air space fluid absorption secondary to inadequate Na+ uptake by the distal lung epithelium that results in fluid-filled lungs at birth with reduced capacity to establish postnatal breathing, and 2) the main stimulus to lung fluid absorption in near-term control fetuses, elevated endogenous epinephrine levels, is not functional in CDH fetuses. PMID- 16214818 TI - Mechanism of ACh-induced asynchronous calcium waves and tonic contraction in porcine tracheal muscle bundle. AB - Stimulation of the tracheal muscle bundle by acetylcholine (ACh) results in the generation of asynchronous repetitive Ca2+ waves (ACW) in intact tracheal smooth muscle (TSM) cells. We showed previously that ACW underlie cholinergic excitation contraction coupling in porcine TSM and that Ca2+ entry through the L-type voltage-gated Ca2+ channel (VGCC) contributes partially to maintenance of the ACW. However, the mechanism of the ACW remains undefined. In this study, we pharmacologically characterized the mechanism of ACh-induced ACW in the intact porcine tracheal muscle bundle. We found that inhibition of receptor-operated channels/store-operated channels (ROC/SOC) by SKF-96365 completely abolished the nifedipine-insensitive component of ACh-mediated ACW and tonic contraction. Blockade of Na+/Ca2+ exchange with KB-R7943 or 2',4'-dichlorobenzamil or removal of extracellular Na+ resulted in nearly complete inhibition of the nifedipine insensitive component of ACh-mediated ACW and tonic contraction. Inhibition of the sarco(endo)plasmic reticulum Ca2+-ATPase by cyclopiazonic acid abolished the ongoing ACW. Application of 2-aminoethoxydiphenyl borate (2-APB) or xestospongin C to inhibit the inositol 1,4,5-trisphosphate-sensitive sarcoplasmic reticulum (SR) Ca2+ release channels produced no effect on ACh-mediated ACW and tonic contraction. However, pretreatment with caffeine or ryanodine inhibited ACh induced ACW. Furthermore, application of procaine or tetracaine prevented the generation and abolished the ongoing ACh-mediated ACW and tonic contraction. Collectively, these results indicate that the ACh-stimulated ACW in porcine TSM are produced by repetitive cycles of Ca2+ release from SR through 2-APB- and xestospongin C-insensitive Ca2+ release channels, and plasmalemmal Ca2+ entry involving reverse-mode Na+/Ca2+ exchange, ROC/SOC, and L-type VGCC is required to refill the SR via SERCA to support the ongoing ACW. PMID- 16214819 TI - Mechanisms of pulmonary edema clearance. AB - The mechanisms of pulmonary edema resolution are different from those regulating edema formation. Absorption of excess alveolar fluid is an active process that involves vectorial transport of Na+ out of alveolar air spaces with water following the Na+ osmotic gradient. Active Na+ transport across the alveolar epithelium is regulated via apical Na+ and chloride channels and basolateral Na-K ATPase in normal and injured lungs. During lung injury, mechanisms regulating alveolar fluid reabsorption are inhibited by yet unclear pathways and can be upregulated by pharmacological means. Better understanding of the mechanisms that regulate edema clearance may lead to therapeutic interventions to improve the ability of lungs to clear fluid, which is of clinical significance. PMID- 16214820 TI - Smaller is not always better: nanotechnology yields nanotoxicology. PMID- 16214821 TI - Herpesviruses: a cofactor in the pathogenesis of idiopathic pulmonary fibrosis? PMID- 16214822 TI - Diesel effects on human health: a question of stress? PMID- 16214823 TI - Stage-specific regulation of respiratory epithelial cell differentiation by Foxa1. AB - Foxa1 is a member of the winged helix family of transcription factors that is expressed in epithelial cells of the conducting airways and in alveolar type II cells of the lung. To determine the role of Foxa1 during lung morphogenesis, histology and gene expression were assessed in lungs from Foxa1-/- gene-targeted mice from embryonic day (E) 16.5 to postnatal day (PN) 13. Deletion of Foxa1 perturbed maturation of the respiratory epithelium at precise times during lung morphogenesis. While dilatation of peripheral lung saccules was delayed in Foxa1 /- mice at E16.5, sacculation was unperturbed later in development (E17.5-E18.5). At PN5, alveolarization was markedly delayed in Foxa1-/- mice; however, by PN13 lung histology was comparable to wild-type controls. Clara cell secretory protein (CCSP), prosurfactant protein (SP)-C, and SP-B protein content and immunostaining were decreased in Foxa1-/- mice between E16.5 and E18.5 but normalized after birth. Timing and sites of expression of thyroid transcription factor-1, Foxj1, and beta-tubulin were unaltered in lungs of Foxa1-/- mice. In vitro, Foxa1 regulated the activity of CCSP and SP-A, SP-B, SP-C, and SP-D promoters as assessed by luciferase reporter assays in HeLa, H441, and MLE15 cells. Although Foxa1 regulates respiratory epithelial differentiation and structural maturation of the lung at precise developmental periods, the delay in maturation is subsequently compensated at times to enable respiratory function and restore normal lung structure after birth. PMID- 16214824 TI - Thioredoxin and dihydrolipoic acid inhibit elastase activity in cystic fibrosis sputum. AB - Excessive neutrophil elastase activity within airways of cystic fibrosis (CF) patients results in progressive lung damage. Disruption of disulfide bonds on elastase by reducing agents may modify its enzymatic activity. Three naturally occurring dithiol reducing systems were examined for their effects on elastase activity: 1) Escherichia coli thioredoxin (Trx) system, 2) recombinant human thioredoxin (rhTrx) system, and 3) dihydrolipoic acid (DHLA). The Trx systems consisted of Trx, Trx reductase, and NADPH. As shown by spectrophotometric assay of elastase activity, the two Trx systems and DHLA inhibited purified human neutrophil elastase as well as the elastolytic activity present in the soluble phase (sol) of CF sputum. Removal of any of the three Trx system constituents prevented inhibition. Compared with the monothiols N-acetylcysteine and reduced glutathione, the dithiols displayed greater elastase inhibition. To streamline Trx as an investigational tool, a stable reduced form of rhTrx was synthesized and used as a single component. Reduced rhTrx inhibited purified elastase and CF sputum sol elastase without NADPH or Trx reductase. Because Trx and DHLA have mucolytic effects, we investigated changes in elastase activity after mucolytic treatment. Unprocessed CF sputum was directly treated with reduced rhTrx, the Trx system, DHLA, or DNase. The Trx system and DHLA did not increase elastase activity, whereas reduced rhTrx treatment increased sol elastase activity by 60%. By contrast, the elastase activity after DNase treatment increased by 190%. The ability of Trx and DHLA to limit elastase activity combined with their mucolytic effects makes these compounds potential therapies for CF. PMID- 16214826 TI - Erythrocyte folate extraction and quantitative determination by liquid chromatography-tandem mass spectrometry: comparison of results with microbiologic assay. AB - BACKGROUND: Erythrocyte folate analysis is an important diagnostic tool to establish folate status or screen for folate deficiency. METHODS: We evaluated conditions that influence the complete hemolysis and deconjugation of folate polyglutamates to folate monoglutamates (FMGs) from whole blood (WB). WB samples were hemolyzed in 10 g/L ascorbic acid at various temperatures (room temperature, 30 degrees C, and 37 degrees C; n = 15) or hemolysate pH values (pH 4.0, 4.7, 5.2; n = 11) and incubated up to 6 h. FMGs and folate diglutamates (FDGs) were analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) and total folate (TF) by microbiologic assay. We investigated delaying hemolysis by freezing WB for 10 days (n = 20). RESULTS: Hemolysates frozen immediately after preparation contained 22%-27% FDGs, depending on hemolysate pH. The proportion of FDGs decreased to <3% after incubation at pH 4.7/37 degrees C for 3 h and did not significantly change on extended incubation up to 5 h. Short-term delayed hemolysis of WB produced results indistinguishable from those of immediate hemolysis. TF results obtained by the microbiologic assay were not different across incubation conditions and agreed with the sum of FMGs and FDGs by LC/MS/MS. The difference between the 2 methods was an insignificant 3% for pH 4.7/37 degrees C for 3 h. CONCLUSIONS: Hemolysate incubation up to 2 h at 37 degrees C is not adequate for full polyglutamate deconjugation. We obtained the highest yield of FMGs with lowest FDG concentrations at pH 4.7/37 degrees C for 3 h. Delaying hemolysis of WB for several days had no negative effect on measurable folate for presumed MTHFR C/C genotype samples. PMID- 16214825 TI - Association of inosine triphosphatase 94C>A and thiopurine S-methyltransferase deficiency with adverse events and study drop-outs under azathioprine therapy in a prospective Crohn disease study. AB - BACKGROUND: Azathioprine (aza) therapy is beneficial in the treatment of inflammatory bowel disease, but 10%-30% of patients cannot tolerate aza therapy because of adverse drug reactions. Thiopurine S-methyltransferase (TPMT) deficiency predisposes to myelotoxicity, but its association with other side effects is less clear. Inosine triphosphatase (ITPA) mutations are other pharmacogenetic polymorphisms possibly involved in thiopurine metabolism and tolerance. METHODS: We analyzed data from a 6-month prospective study including 71 patients with Crohn disease undergoing first-time aza treatment with respect to aza intolerance. Patients were genotyped for common TPMT and ITPA mutations and had pretherapy TPMT activity measured. RESULTS: Early drop-out (within 2 weeks) from aza therapy was associated with ITPA 94C > A [P = 0.020; odds ratio (OR), 4.6; 95% confidence interval (95% CI), 1.2-17.4] and low TPMT activity [<10 nmol/(mL erythrocytes . h); P = 0.007; OR = 5.5; 95% CI, 1.6-19.2]. A high-risk group defined by ITPA 94C > A or TPMT <10 nmol/(mL erythrocytes . h) showed significant association with early drop-out (P = 0.001; OR = 11.3; 95% CI, 2.5 50.0) and all drop-outs (P = 0.002; OR = 4.8; 95% CI, 1.8-13.3). For only drop outs attributable to aza-related side effects (n = 16), there was a significant association with ITPA 94C > A (P = 0.002; OR = 7.8; 95% CI, 2.1-29.1). Time-to event analysis over the 24-week study period revealed a significant association (P = 0.031) between the time to drop-out and ITPA 94C > A mutant allele carrier status. CONCLUSIONS: Patients with ITPA 94C > A mutations or low TPMT activity constitute a pharmacogenetic high-risk group for drop-out from aza therapy. ITPA 94C>A appears to be a promising marker indicating predisposition to aza intolerance. PMID- 16214827 TI - Comparison of the relationships between serum apolipoprotein B and serum lipid distributions. AB - BACKGROUND: Apolipoprotein B (apo B) has been reported to be a better predictor of coronary artery disease than cholesterol indices. The objectives of this study were to evaluate concordances/discordances between cholesterol indices and apo B and to assess the factors that influence them. METHODS: For this study, 11 816 individuals (6965 males, 4851 females), none of whom had a past history of coronary artery disease, were selected from among visitors to the health promotion center at Kangbuk Samsung Hospital between January and December 2002. We assessed concordances between the biochemical indices of atherogenicity and evaluated factors associated with discordances. RESULTS: Apo B and various cholesterol indices were correlated, although concordance fell within the range 47%-56%. Multinomial logistic regression analysis showed an increasing risk of a disproportionately higher apo B than LDL-cholesterol in males, the elderly, smokers, individuals with metabolic syndrome, in those with high HDL-cholesterol or triglyceride (TG) concentrations or larger waist circumferences, and in those with low total cholesterol (TC). CONCLUSIONS: The introduction of apo B to standard lipid profile testing could improve the evaluation of risk factors of coronary artery disease and aid more accurate assessment of the effects of cholesterol-lowering therapy, particularly in males, the elderly, smokers, or in individuals with metabolic syndrome, high HDL-cholesterol, high TGs, larger waist circumferences, or low TC. PMID- 16214828 TI - Monitoring temporary immunodepression by flow cytometric measurement of monocytic HLA-DR expression: a multicenter standardized study. AB - BACKGROUND: Single-center trials have shown that monocytic HLA-DR is a good marker for monitoring the severity of temporary immunodepression after trauma, major surgery, or sepsis. A new test for measuring monocytic HLA-DR is now available. METHODS: We evaluated a new test reagent set for monocytic HLA-DR expression (BD Quantibritetrade mark HLA-DR/Monocyte reagent; Becton Dickinson) in single-laboratory and interlaboratory experiments, assessing preanalytical handling, lyse-no-wash (LNW) vs lyse-wash (LW) values, reference values, and the effect of use of different flow cytometers and different instrument settings on test variance. RESULTS: For preanalytical handling, EDTA anticoagulation, storage on ice as soon as possible, and staining within 4 h after blood collection gave results comparable to values obtained for samples analyzed immediately after collection (mean increase of approximately 4% in monocytic HLA-DR). Comparison of LNW and LW revealed slightly higher results for LNW ( approximately 18% higher for LNW compared with LW; r = 0.982). Comparison of different flow cytometers and instrument settings gave CVs <4%, demonstrating the independence of the test from these variables and suggesting that this method qualifies as a standardized test. CV values from the interlaboratory comparison ranged from 15% (blood sample unprocessed before transport) to 25% (stained and fixed before transport). CONCLUSIONS: For the BD Quantibrite HLA-DR/Monocyte test, preanalytical handling is standardized. Single-laboratory results demonstrated the independence of this test from flow cytometer and instrument settings. Interlaboratory results showed greater variance than single-laboratory values. This interlaboratory variance was partly attributable to the influence of transport and can be reduced by optimization of transport conditions. PMID- 16214829 TI - Direct thrombin inhibitors in acute coronary syndromes: effect in patients undergoing early percutaneous coronary intervention. AB - AIMS: We evaluated the effect of direct thrombin inhibitors (DTIs) in patients undergoing early percutaneous coronary intervention (PCI), using the DTI Trialists' Collaboration database of 35,970 patients from 11 randomized trials of DTIs vs. heparin. METHODS AND RESULTS: We performed a Cox proportional hazards regression analysis with PCI as a time-dependent covariate to assess the independent impact of DTIs according to the performance of early PCI. PCI was performed in 7049 patients in the first 72 h after randomization. In trials in which PCI was not planned, DTIs were associated with a 10% relative risk reduction in death or myocardial infarction at 30 days (HR=0.90, 95% CI: 0.84 0.97). This benefit was found to be greater in patients undergoing early PCI (HR=0.66, 95% CI: 0.48-0.91) than those undergoing non-early PCI (HR=0.94, 95% CI: 0.86-1.03). After adjustment for baseline characteristics and propensity to undergo PCI, the risk of death or myocardial infarction remained lower with DTI (HR=0.62, 95% CI: 0.44-0.89). CONCLUSION: After adjustment for baseline differences and propensity to undergo early PCI, DTIs appeared to be more effective than heparin in reducing death or re-infarction among patients undergoing early PCI. PMID- 16214830 TI - Efficacy of ivabradine, a new selective I(f) inhibitor, compared with atenolol in patients with chronic stable angina. AB - AIMS: Ivabradine, a new I(f) inhibitor which acts specifically on the pacemaker activity of the sinoatrial node, is a pure heart rate lowering agent. Ivabradine has shown anti-ischaemic and anti-anginal activity in a placebo-controlled trial. The objective of this study was to compare the anti-anginal and anti-ischaemic effects of ivabradine and the beta-blocker atenolol. METHODS AND RESULTS: In a double-blinded trial, 939 patients with stable angina were randomized to receive ivabradine 5 mg bid for 4 weeks and then either 7.5 or 10 mg bid for 12 weeks or atenolol 50 mg od for 4 weeks and then 100 mg od for 12 weeks. Patients underwent treadmill exercise tests at randomization (M(0)) and after 4 (M(1)) and 16 (M(4)) weeks of therapy. Increases in total exercise duration (TED) at trough at M(4) were 86.8+/-129.0 and 91.7+/-118.8 s with ivabradine 7.5 and 10 mg, respectively and 78.8+/-133.4 s with atenolol 100 mg. Mean differences (SE) when compared with atenolol 100 mg were 10.3 (9.4) and 15.7 (9.5) s in favour of ivabradine 7.5 and 10 mg (P<0.001 for non-inferiority). TED at M(1) improved by 64.2+/-104.0 s with ivabradine 5 mg and by 60.0+/-114.4 s with atenolol 50 mg (P<0.001 for non inferiority). Non-inferiority of ivabradine was shown at all doses and for all criteria. The number of angina attacks was decreased by two-thirds with both ivabradine and atenolol. CONCLUSION: Ivabradine is as effective as atenolol in patients with stable angina. PMID- 16214831 TI - Catheter ablation treatment in patients with drug-refractory atrial fibrillation: a prospective, multi-centre, randomized, controlled study (Catheter Ablation For The Cure Of Atrial Fibrillation Study). AB - AIMS: We conducted a multi-centre, prospective, controlled, randomized trial to investigate the adjunctive role of ablation therapy to antiarrhythmic drug therapy in preventing atrial fibrillation (AF) relapses in patients with paroxysmal or persistent AF in whom antiarrhythmic drug therapy had already failed. METHODS AND RESULTS: One hundred and thirty seven patients were randomized to ablation and antiarrhythmic drug therapy (ablation group) or antiarrhythmic drug therapy alone (control group). In the ablation group, patients underwent cavo-tricuspid and left inferior pulmonary vein (PV)-mitral isthmus ablation plus circumferential PV ablation. The primary end-point of the study was the absence of any recurrence of atrial arrhythmia lasting >30 s in the 1-year follow-up period, after 1-month blanking period. Three (4.4%) major complications were related to ablation: one patient had a stroke during left atrium ablation, another suffered transient phrenic paralysis, and the third had a pericardial effusion which required pericardiocentesis. After 12 months of follow-up, 63/69 (91.3%) control group patients had at least one AF recurrence, whereas 30/68 (44.1%) (P<0.001) ablation group patients had atrial arrhythmia recurrence (four patients had atrial flutter, 26 patients AF). CONCLUSION: Ablation therapy combined with antiarrhythmic drug therapy is superior to antiarrhythmic drug therapy alone in preventing atrial arrhythmia recurrences in patients with paroxysmal or persistent AF in whom antiarrhythmic drug therapy has already failed. PMID- 16214832 TI - Remodelling of the right ventricle after early pulmonary valve replacement in children with repaired tetralogy of Fallot: assessment by cardiovascular magnetic resonance. AB - AIMS: Correct timing of pulmonary valve replacement (PVR) is crucial for preventing complications of pulmonary regurgitation and right ventricular (RV) dilatation after repair of tetralogy of Fallot. We sought to assess the remodelling of the RV after early PVR in children, using cardiovascular magnetic resonance (CMR). METHODS AND RESULTS: Twenty children with severe pulmonary regurgitation and RV dilatation and mean age 13.9 +/- 3 years underwent CMR evaluation 5.6 +/- 1.8 months before and 5.9 +/- 0.6 months after PVR. PVR was performed when the RV end-diastolic volume exceeded 150 mL/m(2), as measured by CMR. The time interval between primary repair and PVR was 12 +/- 3 years. Post operative CMR demonstrated a significant reduction of the RV end-diastolic volume from 189.8 +/- 33.4 to 108.7 +/- 25.8 mL/m(2) (P < 0.0001), of the RV end systolic volume from 102.4 +/- 27.3 to 58.2 +/- 16.3 mL/m(2) (P < 0.0001), and of the RV mass from 48.7 +/- 12.3 to 35.8 +/- 7.7 g/m(2) (P < 0.0001). The RV ejection fraction did not change significantly. CONCLUSION: Prompt RV remodelling, with reduction of RV volume and mass, is observed after performing PVR if the RV end-diastolic volume exceeds 150 mL/m(2). Early PVR may prevent the detrimental complications of severe pulmonary regurgitation. PMID- 16214833 TI - Bundle-branch block in middle-aged men: risk of complications and death over 28 years. The Primary Prevention Study in Goteborg, Sweden. AB - AIMS: To investigate the long-term fate of men with bundle-branch block (BBB) from a general population sample. METHODS AND RESULTS: Data were derived from 7392 men without a history of myocardial infarction or stroke, born between 1915 and 1925 and investigated between 1970 and 1973. All participants were followed from the date of their baseline examination until 1998. We identified 70 men with right-BBB and 46 men with left-BBB at baseline. In men with right-BBB, there was no increased risk of myocardial infarction, coronary death, heart failure, or all cause mortality during follow-up. The multiple-adjusted hazard ratio for progression to high-degree atrioventricular block was 3.64 (99% confidence interval 0.79-16.72). In men with left-BBB, the hazard ratio for high-degree atrioventricular block was 12.89 (4.13-40.24). However, hazard ratio for all cause mortality was 1.85 (1.15-2.97) when compared with men without BBB, mostly due to outside hospital coronary deaths, whose hazard ratio was 4.22 (1.90-9.34). CONCLUSION: The presence of BBB was strongly associated with future high-degree atrioventricular block that was more pronounced for left-BBB. Men with left-BBB have a substantially increased risk of coronary death, mainly due to sudden death outside the hospital setting. PMID- 16214834 TI - Left is worse than right: the outcome of bundle branch block in middle-aged men. PMID- 16214835 TI - Cancer cachexia. AB - Cancer cachexia is a severe debilitating disorder for which there are currently few therapeutic options. It is driven by the release of pro-inflammatory cytokines and cachectic factors by both host and tumour. Over the past few years, basic science advances have begun to reveal the breadth and complexity of the immunological mechanisms involved, and in the process have uncovered some novel potential therapeutic targets. The effectiveness of thalidomide and eicosapentaenoic acid at attenuating weight loss in clinical trials also provides a further rationale for modulating the immune response. We are now entering an exciting period in cachexia research, and it is likely that the next few years will see effective new biological therapies reach clinical practice. PMID- 16214836 TI - Vascular injury response in mice is dependent on genetic background. AB - Mouse models are employed to unravel the pathophysiology of vascular restenosis. Although much effort has been spent on how to apply an adequate arterial injury, the influence of the genetic background of mice has not yet received sufficient consideration. The study presented herein was designed to demonstrate the influence of the mouse strain on vascular injury response. Mice of a defined background (50% 129 strain and 50% DBA strain) were backcrossed into either the 129 strain or the DBA strain. Male offspring were subjected to a femoral artery injury model by applying an electric current. Morphometric analysis revealed that backcrossing into the 129 strain resulted in a significant (P < 0.001) 17-fold increase in neointima formation (n = 17 mice) compared with backcrossing into the DBA strain (n = 19). The values of neointima area were 9.18 x 10(3) +/- 2.13 x 10(3) and 0.54 x 10(3) +/- 0.39 x 10(3) microm2, respectively. In conjunction, the vessel wall area was enhanced by 1.8-fold (P < 0.001). In contrast, no significant differences were found for the areas of the lumen and the tunica media. Similarly, a significant increase in neointima formation was also found for mice of pure 129 strain compared with pure DBA strain. The results underline the importance of the genetic background for studies on vascular injury response. Furthermore, because the mouse genome of the various strains is well defined, serial testing of the genetic background of mice will provide candidate genes and/or genetic modifiers controlling vascular injury response. PMID- 16214837 TI - Superoxide anion is elevated in sympathetic neurons in DOCA-salt hypertension via activation of NADPH oxidase. AB - Superoxide anion (O2-*) production is elevated in sympathetic ganglion neurons and in the vasculature of hypertensive animals; however, it is not known what enzymatic pathway(s) are responsible for O2-* production. To determine the pathway(s) of O2-* production in sympathetic neurons, we examined the presence of mRNA of NADPH oxidase subunits in sympathetic ganglionic neurons and differentiated PC-12 cells. The mRNAs for NADPH oxidase subunits p47phox, p22phox, gp91phox, and NOX1 were present in sympathetic neurons and PC-12 cells, whereas the NOX4 homologue was present in sympathetic neurons but not PC-12 cells. Freshly dissociated celiac ganglion neurons from normal rats and PC-12 cells produced O2-* when treated with the PKC activator PMA; O2-* production increased by 317% and 254%, respectively. The PMA-evoked increases were reduced by pretreatment with the NADPH oxidase inhibitor apocynin. These findings indicate that NADPH oxidase is the primary source of O2-* in sympathetic ganglion neurons. When celiac ganglia from hypertensive rats were incubated with apocynin, O2-* levels were reduced to the same levels as normotensive animals, indicating that NADPH oxidase activity accounted for the elevated O2-* levels in hypertensive animals. To test this latter finding, we compared NADPH oxidase activity in extracts of prevertebral sympathetic ganglia of DOCA-salt hypertensive rats and sham-operated rats. NADPH oxidase activities were 49.9% and 78.6% higher in sympathetic ganglia of DOCA rats compared with normotensive controls when using beta-NADH and beta-NADPH as substrates, respectively. Thus elevated O2-* levels in hypertension may be a result of the increased activity of NADPH oxidase in postganglionic sympathetic neurons. PMID- 16214838 TI - Effects of selective inhibition of cytochrome P-450 omega-hydroxylases and ischemic preconditioning in myocardial protection. AB - Cytochrome P-450 (CYP) omega-hydroxylases and their arachidonic acid (AA) metabolite, 20-hydroxyeicosatetraenoic acid (20-HETE), produce a detrimental effect on ischemia-reperfusion injury in canine hearts, and the inhibition of CYP omega-hydroxylases markedly reduces myocardial infarct size expressed as a percentage of the area at risk (IS/AAR, %). In this study, we demonstrated that a specific CYP omega-hydroxylase inhibitor, N-methylsulfonyl-12,12-dibromododec-11 enamide (DDMS), markedly reduced 20-HETE production during ischemia-reperfusion and reduced myocardial infarct size compared with control [19.5 +/- 1.0% (control), 9.6 +/- 1.5% (0.40 mg/kg DDMS), 4.0 +/- 2.0% (0.81 mg/kg DDMS), P < 0.01]. In addition, 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid (20-HEDE, a putative 20-HETE antagonist) significantly reduced myocardial infarct size from control [10.3 +/- 1.3% (0.032 mg/kg 20-HEDE) and 5.9 +/- 1.9% (0.064 mg/kg 20-HEDE), P < 0.05]. We further demonstrated that one 5-min period of ischemic preconditioning (IPC) reduced infarct size to a similar extent as that observed with the high doses of DDMS and 20-HEDE, and the higher dose of DDMS given simultaneously with IPC augmented the infarct size reduction [9.9 +/- 2.8% (IPC) to 2.5 +/- 1.4% (0.81 mg/kg DDMS), P < 0.05] to a greater degree than that observed with either treatment alone. These results suggest an important negative role for endogenous CYP omega-hydroxylases and their product, 20-HETE, to exacerbate myocardial injury in canine myocardium. Furthermore, for the first time, this study demonstrates that the effect of IPC and the inhibition of CYP omega-hydroxylase synthesis (DDMS) or its actions (20-HEDE) may have additive effects in protecting the canine heart from ischemia-reperfusion injury. PMID- 16214839 TI - Interaction between AT1 and AT2 receptors during postinfarction left ventricular remodeling. AB - The relative contribution of the angiotensin II type 1 and 2 receptors (AT1-R and AT2-R) in postmyocardial infarction (MI) remodeling remains incompletely understood. We studied five groups of C57Bl/6 mice after 1 h of left anterior descending artery occlusion-reperfusion: 1) wild type, untreated (n = 12); 2) wild type, treated with the AT1-R blocker losartan (10-20 mg.kg(-1).day(-1) in drinking water) from day 1 to day 28 post-MI (n = 10); 3) cardiac overexpression of the AT2-R [AT2-transgenic (TG); n = 14]; 4) AT2-TG treated with losartan (n = 13); and 5) AT2-TG and null for the AT1a-R [AT2-TG/AT1 knockout (KO); n = 10]. Cardiac magnetic resonance imaging (CMR) measured ejection fraction and left ventricular end-diastolic and end-systolic volume (EDVI and ESVI) and mass indexed to weight on days 0, 1, 7, and 28 post-MI. Infarct size was measured on day 1 by late gadolinium-enhanced CMR. Regional myocyte hypertrophy and collagen content were measured on day 28 post-MI. Infarct size was similar among groups. Systolic blood pressure was lowest in AT2-TG/AT1KO. By day 28 post-MI, when corrected for baseline differences, EDVI and ESVI were higher and ejection fraction was lower in wild type than other groups. Ejection fraction was highest and EDVI and mass index were lowest in AT2-TG/AT1KO at day 28. The AT2-TG/AT1KO demonstrated less fibrosis in adjacent regions. Regional myocyte hypertrophy was similar in all groups. The AT1-R and AT2-R are intricately intertwined in post-MI remodeling. Pharmacological blockade of AT1-R is equivalent to AT2-R overexpression in attenuating post-MI remodeling. Genetic knockout of the AT1a-R is additive to AT2-R overexpression, due, at least in part, to blood pressure lowering. PMID- 16214840 TI - Ischemic postconditioning during reperfusion activates Akt and ERK without protecting against lethal myocardial ischemia-reperfusion injury in pigs. AB - Transient episodes of ischemic preconditioning (PC) render myocardium protected against subsequent lethal injury after ischemia and reperfusion. Recent studies indicate that application of short, repetitive ischemia only during the onset of reperfusion after the lethal ischemic event may obtain equivalent protection. We assessed whether such ischemic postconditioning (Postcon) is cardioprotective in pigs by limiting lethal injury. Pentobarbital sodium-anesthetized, open-chest pigs underwent 30 min of complete occlusion of the left anterior descending coronary artery and 3-h reflow. PC was elicited by two cycles of 5-min occlusion plus 10-min reperfusion before the 30-min occlusion period. Postcon was elicited by three cycles of 30-s reperfusion, followed by 30-s reocclusion, after the 30 min occlusion period and before the 3-h reflow. Infarct size (%area-at-risk using triphenyltetrazolium chloride macrochemistry; means +/- SE) after 30 min of ischemia was 26.5 +/- 5.2% (n = 7 hearts/treatment group). PC markedly limited myocardial infarct size (2.8 +/- 1.2%, n = 7 hearts/treatment group, P < 0.05 vs. controls). However, Postcon had no effect on infarct size (37.8 +/- 5.1%, n = 7 hearts/treatment group). Within the subendocardium, Postcon increased phosphorylation of Akt (74 +/- 12%) and ERK1/2 (56 +/- 10%) compared with control hearts subjected only to 30-min occlusion and 15-min reperfusion (P < or = 0.05), and these changes were not different from the response triggered by PC (n = 5 hearts/treatment group). Phosphorylation of downstream p70S6K was also equivalent in PC and Postcon groups. These data do not support the hypothesis that application of 30-s cycles of repetitive ischemia during reperfusion exerts a protective effect on pig hearts subjected to lethal ischemia, but this is not due to a failure to phosphorylate ERK and Akt during early reperfusion. PMID- 16214841 TI - Dual effects of mesenteric lymph isolated from rats with burn injury on contractile function in rat ventricular myocytes. AB - Gut-derived factors in intestinal lymph have been shown to trigger myocardial contractile dysfunction. However, the underlying cellular mechanisms remain unclear. We examined the effects of physiologically relevant concentrations of mesenteric lymph collected from rats with 40% burn injury (burn lymph) on excitation-contraction coupling in rat ventricular myocytes. Burn lymph (0.1-5%), but not control mesenteric lymph from sham-burn animals, induced dual positive and negative inotropic effects depending on the concentrations used. At lower concentrations (<0.5%), burn lymph increased the amplitude of myocyte contraction (1.6 +/- 0.3-fold; n = 12). At higher concentrations (>0.5%), burn lymph initially enhanced myocyte contraction, which was followed by a block of contraction. These effects were partially reversible on washout. The initial positive inotropic effect was associated with a prolongation of action potential duration (measured at 90% repolarization, 2.5 +/- 0.6-fold; n = 10), leading to significant increases in the net Ca2+ influx (1.7 +/- 0.1-fold; n = 8). There were no significant changes in the resting membrane potential. The negative inotropic effect was accompanied by a decrease in the action potential plateau (overshoot decrease by 69 +/- 10%; n = 4) and membrane depolarization. Voltage clamp experiments revealed that the positive inotropic effects of burn lymph were due to an inhibition of the transient outward K+ currents that prolong action potential duration, and the inhibitory effects were due to a concentration dependent inhibition of Ca2+ currents that lead to a reduction of action potential plateau. These burn lymph-induced changes in cardiac myocyte Ca2+ handling can contribute to burn-induced contractile dysfunction and ultimately to heart failure. PMID- 16214842 TI - Selenium prevents diabetes-induced alterations in [Zn2+]i and metallothionein level of rat heart via restoration of cell redox cycle. AB - Intracellular free zinc concentration ([Zn2+]i) is very important for cell functions, and its excessive accumulation is cytotoxic. [Zn2+]i can increase rapidly in cardiomyocytes because of mobilization of Zn2+ from intracellular stores by reactive oxygen species (ROS). Moreover, ROS have been proposed to contribute to direct and/or indirect damage to cardiomyocytes in diabetes. To address these hypotheses, we investigated how elevated [Zn2+]i in cardiomyocytes could contribute to diabetes-induced alterations in intracellular free calcium concentration ([Ca2+]i). We also investigated its relationship to the changes of metallothionein (MT) level of the heart. Cardiomyocytes from normal rats loaded with fura-2 were used to fluorometrically measure resting [Zn2+]i (0.52 +/- 0.06 nM) and [Ca2+]i (26.53 +/- 3.67 nM). Fluorescence quenching by the heavy metal chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine was used to quantify [Zn2+]i. Our data showed that diabetic cardiomyocytes exhibited significantly increased [Zn2+]i (0.87 +/- 0.05 nM ) and [Ca2+]i (49.66 +/- 9.03 nM), decreased levels of MT and reduced glutathione, increased levels of lipid peroxidation and nitric oxide products, and decreased activities of superoxide dismutase, glutathione reductase, and glutathione peroxidase. Treatment (4 wk) of diabetic rats with sodium selenite (5 micromol.kg body wt(-1).day(-1)) prevented these defects induced by diabetes. A comparison of present data with previously observed beneficial effects of selenium treatment on diabetes-induced contractile dysfunction of the heart can suggest that an increase in [Zn2+]i may contribute to oxidant-induced alterations of excitation-contraction coupling in diabetes. In addition, we showed that oxidative stress is involved in the etiology of diabetes induced downregulation of heart function via depressed endogenous antioxidant defense mechanisms. PMID- 16214843 TI - Sex-specific differences in cerebral arterial myogenic tone in hypertensive and normotensive rats. AB - Cerebral blood flow (CBF) is maintained constant despite changes in systemic blood pressure (BP) through multiple mechanisms of autoregulation such as vascular myogenic reactivity. Our aim was to determine myogenic characteristics of cannulated middle cerebral arteries (MCA) in male and female stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto rats (WKY) at 12 wk of age under pressurised no-flow conditions. MCA pressure-diameter relationships (20 200 mmHg) were constructed in active (with calcium) and passive (without calcium) conditions, and myogenic and mechanical properties were determined. Myogenic reactivity in WKY (P < 0.05) and SHRSP (P < 0.05) males was impaired compared with their female counterparts. Comparison of SHRSP with WKY in males revealed similar myogenic reactivity, but in females SHRSP exhibited augmented myogenic reactivity (P < 0.05). In both sexes, myogenic tone yielded at lower pressure in SHRSP compared with WKY vessels (120-140 vs. 140-180 mmHg). Stress-strain relationships and elastic moduli in WKY rats showed that vessels were stiffer in females than in males. Conversely, in SHRSP, male vessels were stiffer than female vessels. Comparison of strains in males indicated that stiffness was increased in SHRSP compared with WKY vessels, whereas the converse was observed in females. These findings demonstrate that MCA myogenic and distensibility characteristics exhibit significant sex- and strain-dependent differences. Inappropriate myogenic adaptation and augmented vascular stiffness, particularly in male SHRSP, are potential limiting factors in blood flow autoregulation and may increase the predisposition for stroke-related cerebrovascular events. PMID- 16214844 TI - Complement activation-related cardiac anaphylaxis in pigs: role of C5a anaphylatoxin and adenosine in liposome-induced abnormalities in ECG and heart function. AB - Cardiac anaphylaxis is a severe, life-threatening manifestation of acute hypersensitivity reactions to allergens and drugs. Earlier studies highlighted an amplifying effect of locally applied C5a on the process; however, the role of systemic complement (C) activation with C5a liberation in blood has not been explored to date. In the present study, we used the porcine liposome-induced cardiopulmonary distress model for 1) characterizing and quantifying peripheral C activation-related cardiac dysfunction; 2) exploring the role of C5a in cardiac abnormalities and therapeutic potential of C blockage by soluble C receptor type 1 (sCR1) and an anti-C5a antibody (GS1); and 3) elucidating the role of adenosine and adenosine receptors in paradoxical bradycardia, one of the symptoms observed in this model. Pigs were injected intravenously with different liposomes [Doxil and multilamellar vesicles (MLV)], zymosan, recombinant human (rhu) C5a, and adenosine, and the ensuing hemodynamic and cardiac changes (hypotension, tachy- or bradycardia, arrhythmias, ST-T changes, ventricular fibrillation, and arrest) were quantified by ranking on an arbitrary scale [cardiac abnormality score (CAS)]. There was significant correlation between CAS and C5a production by liposomes in vitro, and the liposome-induced cardiac abnormalities were partially or fully reproduced with zymosan, rhuC5a, adenosine, and the selective adenosine A1 receptor agonist cyclopentyl-adenosine. The use of C nonactivator liposomes or pretreatment of pigs with sCR1 or GS1 attenuated the abnormalities. The selective A1 blocker cyclopentyl-xanthine inhibited bradycardia without influencing hypotension, whereas the A(2) blocker 4-(2-[7-amino-2-(2 furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM-24135) had no such effect. These data suggest that 1) systemic C activation can underlie cardiac anaphylaxis, 2) C5a plays a causal role in the reaction, 3) adenosine action via A1 receptors may explain paradoxical bradycardia, and 4) inhibition of C5a formation or action or of A1-receptor function may alleviate the acute cardiotoxicity of liposomal drugs and other intravenous agents that activate C. PMID- 16214845 TI - Cardiac sympathetic neuroprotective effect of desipramine in tachycardia-induced cardiomyopathy. AB - Cardiac sympathetic transmitter stores are reduced in the failing heart. In this study, we proposed to investigate whether the reduction of cardiac sympathetic neurotransmitters was associated with increased interstitial norepinephrine (NE) and reactive oxygen species in congestive heart failure (CHF), using a microdialysis technique and salicylate to detect .OH generation. Rabbits with and without rapid ventricular pacing (340 beats/min) were randomized to receive desipramine (10 mg/day) or placebo for 8 wk. Rapid pacing produced left ventricular dilation and systolic dysfunction. The failing myocardium also showed reduced tissue contents of NE and tyrosine hydroxylase protein and activity. In contrast, myocardial interstitial NE was increased in CHF (0.89 +/- 0.11 ng/ml) compared with the sham-operated animals (0.26 +/- 0.03 ng/ml). In addition, cardiac oxidative stress was increased in CHF animals as measured by myocardial interstitial .OH radical, tissue oxidized glutathione, and oxidized mitochondrial DNA. Desipramine treatment produced significant NE uptake inhibition as evidence by an exaggerated pressor response and a greater increase of myocardial interstitial NE in response to intravenous NE infusion but no significant effects on cardiac function or hemodynamics in sham-operated or CHF animals. However, desipramine treatment attenuated the reductions of tissue NE and tyrosine hydroxylase protein and activity in CHF. Desipramine also prevented the reduction of tyrosine hydroxylase produced by NE in PC12 cells. Thus the reduction of cardiac sympathetic neurotransmitters is related to the increased interstitial NE and tissue oxidative stress in CHF. Also, normal neuronal uptake of NE is required for NE or its oxidized metabolites to exert their neurotoxic effects. PMID- 16214846 TI - Functional and structural remodeling of the myocardial microvasculature in early experimental hypertension. AB - Advanced hypertension (HT), associated with left ventricular hypertrophy (LVH), impairs myocardial microvascular function and structure and leads to increased myocardial hypoxia and growth factor activation. However, the effect of HT on microvascular architecture and its relation to microvascular function, before the development of LVH (early HT), remains unclear. By way of method, pigs were studied after 12 wk of renovascular HT (n = 7) or control (n = 7) animals. Myocardial microvascular function (blood volume and blood flow at baseline and in response to adenosine) was assessed by using electron beam computed tomography (CT). Microvascular architecture was subsequently studied ex vivo using micro-CT, and microvessels (diameter, <500 microm) were counted in situ in three dimensional images (40-microm on-a-side cubic voxels). Myocardial expression of vascular endothelial growth factor, basic fibroblast growth factor, and hypoxia inducible factor-1alpha were also measured. By way of results, left ventricular muscle mass was similar between the groups. The blood volume response to intravenous adenosine was attenuated in HT animals compared with normal animals (+7.4 +/- 17.0 vs. +46.2 +/- 12.3% compared with baseline, P = 0.48 and P = 0.01, respectively). Microvascular spatial density in HT animals was significantly elevated compared with normal animals (246 +/- 26 vs. 125 +/- 20 vessels/cm2, P < 0.05) and correlated inversely with the blood volume response to adenosine. Growth factors expression was increased in HT animals compared with control animals. In conclusion, early HT elicits changes in myocardial microvascular architecture, which are associated with microvascular dysfunction and precede changes in muscle mass. These observations underscore the direct and early effects of HT on the myocardial vasculature. PMID- 16214847 TI - Dependence of acetylcholine and ADP dilation of pial arterioles on heme oxygenase after transfusion of cell-free polymeric hemoglobin. AB - Polymers of cell-free hemoglobin have been designed for clinical use as oxygen carriers, but limited information is available regarding their effects on vascular regulation. We tested the hypothesis that the contribution of heme oxygenase (HO) to acetylcholine-evoked dilation of pial arterioles is upregulated 2 days after polymeric hemoglobin transfusion. Dilator responses to acetylcholine measured by intravital microscopy in anesthetized cats were blocked by superfusion of the HO inhibitor tin protoporphyrin-IX (SnPPIX) in a group that had undergone exchange transfusion with hemoglobin 2 days earlier but not in surgical sham and albumin-transfused groups. However, immunoblots from cortical brain homogenates did not reveal changes in expression of the inducible isoform HO1 or the constitutive isoform HO2 in the hemoglobin-transfused group. To test whether the inhibitory effect of SnPPIX was present acutely after hemoglobin transfusion, responses were measured within an hour of completion of the exchange transfusion. In control and albumin-transfused groups, acetylcholine responses were unaffected by SnPPIX but were blocked by addition of the nitric oxide synthase inhibitor N(omega)-nitro-l-arginine (l-NNA) to the superfusate. In hemoglobin-transfused groups, the acetylcholine response was blocked by either SnPPIX or l-NNA alone. The effect of another HO inhibitor, chromium mesoporphyrin (CrMP), was tested on ADP, another endothelial-dependent dilator, in anesthetized rats. Pial arteriolar dilation to ADP was unaffected by CrMP in controls but was attenuated 62% by CrMP in rats transfused with hemoglobin. It is concluded that 1) polymeric hemoglobin transfusion acutely upregulates the contribution of HO to acetylcholine-induced dilation of pial arterioles in cats, 2) this upregulation persists 2 days after transfusion when 95% of the hemoglobin is cleared from the circulation, and 3) this acute upregulation of HO signaling is ubiquitous in that similar effects were observed with a different endothelial-dependent agonist (i.e., ADP) in a another species (rat). PMID- 16214848 TI - Optical mapping of late myocardial infarction in rats. AB - Late myocardial infarction (MI) is associated with ventricular arrhythmias and sudden cardiac death. The exact mechanistic relationship between abnormal cellular electrophysiology, conduction abnormalities, and arrhythmogenesis associated with late MI is not completely understood. We report a novel, rapid dye superfusion technique to enable whole heart, high-resolution optical mapping of late MI. Optical mapping of action potentials was performed in normal rats and rats with anterior MI 7 days after left anterior descending artery ligation. Hearts from normal rats exhibited normal action potentials and impulse conduction. With the use of programmed stimulation to assess arrhythmia inducibility, 29% of hearts with late MI had inducible sustained ventricular tachycardia, compared with 0% in normal rats. A causal relationship between the site of infarction, abnormal action potential conduction (i.e., block and slow conduction), and arrhythmogenesis was observed. Optical mapping techniques can be used to measure high-resolution action potentials in a whole heart model of late MI. This experimental model reproduces many of the electrophysiological characteristics (i.e., conduction slowing, block, and ventricular tachycardia) associated with MI in patients. Importantly, the results of this study can enhance our ability to understand the interplay between cellular heterogeneity, conduction abnormalities, and arrhythmogenesis associated with MI. PMID- 16214850 TI - Functional characterization and haplotype analysis of polymorphisms in the human equilibrative nucleoside transporter, ENT2. AB - The equilibrative nucleoside transporter 2 (ENT2; SLC29A2) is a bidirectional transporter that is involved in the disposition of naturally occurring nucleosides as well as a variety of anticancer and antiviral nucleoside analogs. The goal of the current study was to evaluate the function of genetic variants in ENT2 in cellular assays and to determine the haplotype structure of the coding and flanking intronic region of the gene. As part of a large study focused on genetic variation in membrane transporters (Leabman et al., 2003), DNA samples from ethnically diverse populations (100 African-Americans, 100 European Americans, 30 Asians, 10 Mexicans, and 7 Pacific Islanders) were screened for variants in membrane transporters, including SLC29A2. Fourteen polymorphic sites in SLC29A2 were found, including 11 in the coding region. Five protein-altering variants were identified: three nonsynonymous variants, and two deletions. Each of the protein-altering variants was found at a very low frequency, occurring only once in the sample population. The nonsynonymous variants and the deletions were constructed via site-directed mutagenesis and were subsequently characterized in Xenopus laevis oocytes. All variants were able to take up inosine with the exception of ENT2-Delta845-846, which resulted in a frameshift mutation that prematurely truncated the protein. ENT2 showed very infrequent variation compared with most other transporter proteins studied, and it was found that five haplotypes were sufficient to describe the entire sample set. The low overall genetic diversity in SLC29A2 makes it unlikely that variation in the coding region contributes significantly to clinically observed differences in drug response. PMID- 16214849 TI - Renal interstitial fluid ATP responses to arterial pressure and tubuloglomerular feedback activation during calcium channel blockade. AB - A close relationship between changes in renal interstitial fluid (RIF) ATP concentrations and renal autoregulatory or tubuloglomerular feedback (TGF) dependent changes in renal vascular resistance (RVR) has been demonstrated, but it has not been determined whether the changes in RIF ATP are a consequence or the cause of the changes in RVR. The present study was performed in anesthetized dogs to assess the changes in RIF ATP following changes in renal arterial pressure (RAP) or stimulation of the TGF mechanism under conditions where changes in RVR were prevented by nifedipine, a calcium channel blocker. RIF ATP levels were measured by using microdialysis probes. Intra-arterial infusion of nifedipine (0.36 microg x kg(-1) x min(-1)) increased renal blood flow (RBF: from 4.49 +/- 0.27 to 5.34 +/- 0.39 ml x min(-1) x g(-1)) and glomerular filtration rate (GFR: from 0.84 +/- 0.07 to 1.09 +/- 0.11 ml x min(-1) x g(-1)). Under conditions of nifedipine infusion, autoregulatory adjustments in RBF, GFR, and RVR were not observed during stepwise reductions in RAP within the autoregulatory range (from 135 +/- 7 to 76 +/- 1 mmHg, n = 7). Furthermore, stimulation of the TGF mechanism with intra-arterial infusion of acetazolamide (100 microg x kg(-1) x min(-1)) did not alter RBF, GFR, and RVR (n = 7). During treatment with nifedipine, RIF ATP levels were significantly decreased in response to reductions in RAP (10.7 +/- 0.7, 5.8 +/- 0.7 and 2.8 +/- 0.3 nmol/l at 135 +/- 7, 101 +/- 4, and 76 +/- 1 mmHg, n = 7) and increased by acetazolamide infusion (from 8.8 +/- 0.8 to 17.0 +/- 1.8 nmol/l, n = 7). These results are similar to those that occurred in dogs not treated with nifedipine and thus demonstrate that the changes in RIF ATP can occur in the absence of autoregulatory or TGF-mediated changes in RVR. The data provide further support to the hypothesis that RIF ATP contributes to adjustments in RVR associated with renal autoregulation and changes in activity of the TGF mechanism. PMID- 16214851 TI - Role of human cyclooxygenase-2 in the bioactivation of dapsone and sulfamethoxazole. AB - Sulfamethoxazole (SMX) and dapsone (4,4'-diaminodiphenylsulfone, DDS) are believed to mediate their adverse effects subsequent to bioactivation to their respective arylhydroxylamine and arylnitroso metabolites, resulting in covalent adduct formation with intracellular proteins. Various bioactivating enzymes, such as cytochromes P450 and myeloperoxidase, have been shown to be capable of catalyzing the N-oxidation of these compounds. We assessed the role of human cyclooxygenase-2 (COX-2) in the metabolism and subsequent adduct formation of DDS and SMX using recombinant human COX-2. Using an adduct-specific enzyme-linked immunosorbent assay, we found that the complete enzyme system gave rise to covalent adducts. However, the nonspecific COX inhibitor indomethacin did not reduce the amount of covalent adduct formed. Formation of the arylhydroxylamine metabolites was demonstrated via high performance liquid chromatography coupled with UV absorption. Metabolite formation was found to be secondary to the H2O2 in the incubation mixture and was not enzyme-mediated. Hence, COX-2 does not play a direct role in the bioactivation of these parent drugs to their arylhydroxylamine metabolites. PMID- 16214853 TI - Optical clearing for improved contrast in second harmonic generation imaging of skeletal muscle. AB - Using second harmonic generation (SHG) imaging microscopy, we have examined the effect of optical clearing with glycerol to achieve greater penetration into specimens of skeletal muscle tissue. We find that treatment with 50% glycerol results in a 2.5-fold increase in achievable SHG imaging depth. Signal processing analyses using fast Fourier transform and continuous wavelet transforms show quantitatively that the periodicity of the sarcomere structure is unaltered by the clearing process and that image quality deep in the tissue is improved with clearing. Comparison of the SHG angular polarization dependence also shows no change in the supramolecular organization of acto-myosin complexes. By contrast, identical treatment of mouse tendon (collagen based) resulted in a strong decrease in SHG response. We suggest that the primary mechanism of optical clearing in muscle with glycerol treatment results from the reduction of cytoplasmic protein concentration and concomitant decrease in the secondary inner filter effect on the SHG signal. The lack of glycerol concentration dependence on the imaging depth indicates that refractive index matching plays only a minor role in the optical clearing of muscle. SHG and optical clearing may provide an ideal mechanism to study physiology in highly scattering skeletal or cardiac muscle tissue with significantly improved depth of penetration and achievable imaging depth. PMID- 16214852 TI - Mechanisms of excitation-contraction coupling in an integrative model of the cardiac ventricular myocyte. AB - It is now well established that characteristic properties of excitation contraction (EC) coupling in cardiac myocytes, such as high gain and graded Ca(2+) release, arise from the interactions that occur between L-type Ca(2+) channels (LCCs) and nearby ryanodine-sensitive Ca(2+) release channels (RyRs) in localized microdomains. Descriptions of Ca(2+)-induced Ca(2+) release (CICR) that account for these local mechanisms are lacking from many previous models of the cardiac action potential, and those that do include local control of CICR are able to reconstruct properties of EC coupling, but require computationally demanding stochastic simulations of approximately 10(5) individual ion channels. In this study, we generalize a recently developed analytical approach for deriving simplified mechanistic models of CICR to formulate an integrative model of the canine cardiac myocyte which is computationally efficient. The resulting model faithfully reproduces experimentally measured properties of EC coupling and whole cell phenomena. The model is used to study the role of local redundancy in L-type Ca(2+) channel gating and the role of dyad configuration on EC coupling. Simulations suggest that the characteristic steep rise in EC coupling gain observed at hyperpolarized potentials is a result of increased functional coupling between LCCs and RyRs. We also demonstrate mechanisms by which alterations in the early repolarization phase of the action potential, resulting from reduction of the transient outward potassium current, alters properties of EC coupling. PMID- 16214854 TI - Cosegregation of permeability and single-channel conductance in chimeric connexins. AB - The physiological function of gap junction channels goes well beyond their initially discovered role in electrical synchronization of excitable cells. In most tissues, gap junction cells facilitate the exchange of second messengers and metabolites between cells. To test which parts of the channels formed by connexins determine the exclusion limit for the transit of molecules in the size range of second messengers and metabolites a domain exchange approach was used in combination with an accessibility assay for nonelectrolytes and flux measurements. The experimental results suggest that two open hemichannel forming connexins, Cx46 and Cx32E(1)43, differ in accessibility and permeability. Sucrose is at the exclusion limit for Cx46 channels whereas sorbitol is at the exclusion limit for Cx32E(1)43 channels. In chimeras between these connexins, where the first transmembrane segment M1 is exchanged, the exclusion limits correlate with those of the M1 donor. The same segregation was found in a separate study for the unitary conductance of the channels. Thus, conductance and permeability/accessibility of the channels cosegregate with M1. PMID- 16214855 TI - Conductance of connexin hemichannels segregates with the first transmembrane segment. AB - Gap junction channels are intercellular channels that mediate the gated transfer of molecules between adjacent cells. To identify the domain determining channel conductance, the first transmembrane segment (M1) was reciprocally exchanged between Cx46 and Cx32E(1)43. The resulting chimeras exhibited conductances similar to that of the respective M1 donor. Furthermore, a chimera with the carboxy-terminal half of M1 in Cx46 replaced by that of Cx32 exhibited a conductance similar to that of Cx32E(1)43, whereas the chimera with only the amino-terminal half of M1 replaced retained the unitary conductance of wild-type Cx46. Extending the M1 domain swapping to other connexins by replacing the carboxy-terminal half of M1 in Cx46 with that of Cx37 yielded a chimera channel with increased unitary conductance close to that of Cx37. Furthermore, a point mutant of Cx46, with leucine substituted by glycine in position 35, displayed a conductance much larger than that of the wild type. Thus, the M1 segment, especially the second half, contains important determinants of conductance of the connexin channel. PMID- 16214857 TI - Determination of RNA orientation during translocation through a biological nanopore. AB - We investigate single-molecule electrophoretic translocation of A(50), C(50), A(25)C(50), and C(50)A(25) RNA molecules through the alpha-hemolysin transmembrane protein pore. We observe pronounced bilevel current blockages during translocation of A(25)C(50) and C(50)A(25) molecules. The two current levels observed during these bilevel blockages are very similar to the characteristic current levels observed during A(50) and C(50) translocation. From the temporal ordering of the two levels within the bilevel current blockages, we infer whether individual A(25)C(50) and C(50)A(25) molecules pass through the pore in a 3'-->5' or 5'-->3' orientation. Correlation between the level of current obstruction and the inferred A(25)C(50) or C(50)A(25) orientation indicates that 3'-->5' translocation of a poly C segment causes a significantly deeper current obstruction than 5'-->3' translocation. Our analysis also suggests that the 3' ends of C(50) and A(25)C(50) RNA molecules are more likely to initiate translocation than the 5' ends. Orientation dependent differences in a smaller current blockage that immediately precedes many translocation events suggest that this blockage also contains information about RNA orientation during translocation. These findings emphasize that the directionality of polynucleotide molecules is an important factor in translocation and demonstrate how structure within ionic current signals can give new insights into the translocation process. PMID- 16214856 TI - A Brownian dynamics study of the interaction of Phormidium cytochrome f with various cyanobacterial plastocyanins. AB - Brownian dynamics simulations were used to study the role of electrostatic forces in the interactions of cytochrome f from the cyanobacterium Phormidium laminosum with various cyanobacterial plastocyanins. Both the net charge on the plastocyanin molecule and the charge configuration around H92 (H87 in higher plants) are important in determining the interactions. Those plastocyanins (PCs) with a net charge more negative than -2.0, including those from Synechococcus sp. PCC7942, Synechocystis sp. 6803, and P. laminosum showed very little complex formation. On the other hand, complex formation for those with a net charge more positive than -2.0 (including Nostoc sp. PCC7119 and Prochlorothrix hollandica) as well as Nostoc plastocyanin mutants showed a linear dependence of complex formation upon the net charge on the plastocyanin molecule. Mutation of charged residues on the surface of the PC molecules also affected complex formation. Simulations involving plastocyanin mutants K35A, R93A, and K11A (when present) showed inhibition of complex formation. In contrast, D10A and E17A mutants showed an increase in complex formation. All of these residues surround the H92 (H87 in higher plant plastocyanins) ligand to the copper. An examination of the closest electrostatic contacts shows that these residues interact with D63, E123, R157, D188, and the heme on Phormidium cytochrome f. In the complexes formed, the long axis of the PC molecule lies perpendicular to the long axis of cytochrome f. There is considerable heterogeneity in the orientation of plastocyanin in the complexes formed. PMID- 16214858 TI - Mechanical resistance of proteins explained using simple molecular models. AB - Recent experiments have demonstrated that proteins unfold when two atoms are mechanically pulled apart, and that this process is different to when heated or when a chemical denaturant is added to the solution. Experiments have also shown that the response of proteins to external forces is very diverse, some of them being "hard," and others "soft." Mechanical resistance originates from the presence of barriers on the energy landscape; together, experiment and simulation have demonstrated that unfolding occurs through alternative pathways when different pairs of atoms undergo mechanical extension. Here we use simulation to probe the mechanical resistance of six structurally diverse proteins when pulled in different directions. For this, we use two very different models: a detailed, transferable one, and a coarse-grained, structure-based one. The coarse-grained model gives results that are surprisingly similar to the detailed one and qualitatively agree with experiment; i.e., the mechanical resistance of different proteins or of a single protein pulled in different directions can be predicted by simulation. The results demonstrate the importance of pulling direction relative to the local topology in determining mechanical stability, and rationalize the effect of the location of importation/degradation tags on the rates of mitochondrial import or protein degradation in vivo. PMID- 16214859 TI - Transmembrane peptides stabilize inverted cubic phases in a biphasic length dependent manner: implications for protein-induced membrane fusion. AB - WALP peptides consist of repeating alanine-leucine sequences of different lengths, flanked with tryptophan "anchors" at each end. They form membrane spanning alpha-helices in lipid membranes, and mimic protein transmembrane domains. WALP peptides of increasing length, from 19 to 31 amino acids, were incorporated into N-monomethylated dioleoylphosphatidylethanolamine (DOPE-Me) at concentrations up to 0.5 mol % peptide. When pure DOPE-Me is heated slowly, the lamellar liquid crystalline (L(alpha)) phase first forms an inverted cubic (Q(II)) phase, and the inverted hexagonal (H(II)) phase at higher temperatures. Using time-resolved x-ray diffraction and slow temperature scans (1.5 degrees C/h), WALP peptides were shown to decrease the temperatures of Q(II) and H(II) phase formation (T(Q) and T(H), respectively) as a function of peptide concentration. The shortest and longest peptides reduced T(Q) the most, whereas intermediate lengths had weaker effects. These findings are relevant to membrane fusion because the first step in the L(alpha)/Q(II) phase transition is believed to be the formation of fusion pores between pure lipid membranes. These results imply that physiologically relevant concentrations of these peptides could increase the susceptibility of biomembrane lipids to fusion through an effect on lipid phase behavior, and may explain one role of the membrane-spanning domains in the proteins that mediate membrane fusion. PMID- 16214860 TI - Structural characterization of apomyoglobin self-associated species in aqueous buffer and urea solution. AB - The biophysical characterization of nonfunctional protein aggregates at physiologically relevant temperatures is much needed to gain deeper insights into the kinetic and thermodynamic relationships between protein folding and misfolding. Dynamic and static laser light scattering have been employed for the detection and detailed characterization of apomyoglobin (apoMb) soluble aggregates populated at room temperature upon dissolving the purified protein in buffer at pH 6.0, both in the presence and absence of high concentrations of urea. Unlike the beta-sheet self-associated aggregates previously reported for this protein at high temperatures, the soluble aggregates detected here have either alpha-helical or random coil secondary structure, depending on solvent and solution conditions. Hydrodynamic diameters range from 80 to 130 nm, with semiflexible chain-like morphology. The combined use of low pH and high urea concentration leads to structural unfolding and complete elimination of the large aggregates. Even upon starting from this virtually monomeric unfolded state, however, protein refolding leads to the formation of severely self-associated species with native-like secondary structure. Under these conditions, kinetic apoMb refolding proceeds via two parallel routes: one leading to native monomer, and the other leading to a misfolded and heavily self-associated state bearing native-like secondary structure. PMID- 16214861 TI - A novel method for detergent concentration determination. AB - A fast and precise method for detergent concentration determination is presented. (Patent applications for the method described here have been submitted (EP05011904 and US60/702,261). Depending on the interest of the scientific community, the system will be commercialized. (For further information contact Herve-W. Remigy at the e-mail address below.) A small droplet of the detergent solution is deposited on a piece of Parafilm M and side views are recorded by two orthogonally arranged TV cameras. The droplet contours are then approximated by ellipses to determine the contact angles. Comparison of the observed contact angle values to calibrated standard curves of known detergent concentrations gives the concentration of the detergent assessed. A range of commonly used detergents was studied to demonstrate the reproducibility and precision of this simple method. As a first application, the detergent binding capacity of the Escherichia coli galactose/proton symporter (GalP) was assessed. Aggregation of GalP was observed when <260 +/- 5 dodecyl-beta,D-maltoside molecules were bound to one GalP molecule. These measurements document the efficacy of the drop-shape based detergent concentration determination described. PMID- 16214862 TI - Quantification of calcium entry at the T-tubules and surface membrane in rat ventricular myocytes. AB - The action potential of cardiac ventricular myocytes is characterized by its long duration, mainly due to Ca flux through L-type Ca channels. Ca entry also serves to trigger the release of Ca from the sarcoplasmic reticulum. The aim of this study was to investigate the role of cell membrane invaginations called transverse (T)-tubules in determining Ca influx and action potential duration in cardiac ventricular myocytes. We used the whole cell patch clamp technique to record electrophysiological activity in intact rat ventricular myocytes (i.e., from the T-tubules and surface sarcolemma) and in detubulated myocytes (i.e., from the surface sarcolemma only). Action potentials were significantly shorter in detubulated cells than in control cells. In contrast, resting membrane potential and action potential amplitude were similar in control and detubulated myocytes. Experiments under voltage clamp using action potential waveforms were used to quantify Ca entry via the Ca current. Ca entry after detubulation was reduced by approximately 60%, a value similar to the decrease in action potential duration. We calculated that Ca influx at the T-tubules is 1.3 times that at the cell surface (4.9 vs. 3.8 micromol/L cytosol, respectively) during a square voltage clamp pulse. In contrast, during a cardiac action potential, Ca entry at the T-tubules is 2.2 times that at the cell surface (3.0 vs. 1.4 micromol/L cytosol, respectively). However, more Ca entry occurs per microm(2) of junctional membrane at the cell surface than in the T-tubules (in nM/microm(2): 1.43 vs. 1.06 during a cardiac action potential). This difference is unlikely to be due to a difference in the number of Ca channels/junction at each site because we estimate that the same number of Ca channels is present at cell surface and T tubule junctions ( approximately 35). This study provides the first evidence that the T-tubules are a key site for the regulation of action potential duration in ventricular cardiac myocytes. Our data also provide the first direct measurements of T-tubular Ca influx, which are consistent with the idea that cardiac excitation-contraction coupling largely occurs at the T-tubule dyadic clefts. PMID- 16214864 TI - Simulation study of the contribution of oligomer/oligomer binding to capsid assembly kinetics. AB - The process by which hundreds of identical capsid proteins self-assemble into icosahedral structures is complex and poorly understood. Establishing constraints on the assembly pathways is crucial to building reliable theoretical models. For example, it is currently an open question to what degree overall assembly kinetics are dominated by one or a few most efficient pathways versus the enormous number theoretically possible. The importance of this question, however, is often overlooked due to the difficulties of addressing it in either theoretical or experimental practice. We apply a computer model based on a discrete-event simulation method to evaluate the contributions of nondominant pathways to overall assembly kinetics. This is accomplished by comparing two possible assembly models: one allowing growth to proceed only by the accretion of individual assembly subunits and the other allowing the binding of sterically compatible assembly intermediates any sizes. Simulations show that the two models perform almost identically under low binding rate conditions, where growth is strongly nucleation-limited, but sharply diverge under conditions of higher association rates or coat protein concentrations. The results suggest the importance of identifying the actual binding pattern if one is to build reliable models of capsid assembly or other complex self-assembly processes. PMID- 16214863 TI - Critical structure-function determinants within the N-terminal region of pulmonary surfactant protein SP-B. AB - Surfactant protein SP-B is absolutely required for the surface activity of pulmonary surfactant and postnatal lung function. The results of a previous study indicated that the N-terminal segment of SP-B, comprising residues 1-9, is specifically required for surface activity, and suggested that prolines 2, 4, and 6 as well as tryptophan 9, may constitute essential structural motifs for protein function. In this work, we assessed the role of these two motifs in promoting the formation and maintenance of surface-active films. Three synthetic peptides were synthesized including a peptide corresponding to the N-terminal 37 amino acids of native SP-B and two variants in which prolines 2, 4, 6, or tryptophan 9 were substituted by alanines. All three synthetic peptides were surface-active, as expected from their amphipathic structure. The peptides were also able to insert into dipalmitoylphosphatidylcholine/palmitoyloleoylphosphatidylglycerol (7:3 w/w ratio) monolayers preformed at pressures >30 mN/m, indicating that they perturb and insert into membranes. Substitution of alanine for tryptophan at position 9 significantly decreased both the rate of adsorption/insertion of the peptide into the interface and reinsertion of surface-active material excluded from the film during successive compression-expansion cycles. Substitution of alanines for prolines at positions 2, 4, and 6 did not produce substantial changes in the rate of adsorption/insertion; however, reinsertion of surface-active material into the expanding interface film was not as effective as in the presence of the nativelike peptide. These results suggest that W9 is critical for optimal interface affinity, whereas prolines may promote a conformation that facilitates rapid insertion of the peptide into phospholipid monolayers compressed to the highest pressures during compression-expansion cycling. PMID- 16214865 TI - In silico studies of the African swine fever virus DNA polymerase X support an induced-fit mechanism. AB - The African swine fever virus DNA polymerase X (pol X), a member of the X family of DNA polymerases, is thought to be involved in base excision repair. Kinetics data indicate that pol X catalyzes DNA polymerization with low fidelity, suggesting a role in viral mutagenesis. Though pol X lacks the fingers domain that binds the DNA in other members of the X family, it binds DNA tightly. To help interpret details of this interaction, molecular dynamics simulations of free pol X at different salt concentrations and of pol X bound to gapped DNA, in the presence and in the absence of the incoming nucleotide, are performed. Anchors for the simulations are two NMR structures of pol X without DNA and a model of one NMR structure plus DNA and incoming nucleotide. Our results show that, in its free form, pol X can exist in two stable conformations that interconvert to one another depending on the salt concentration. When gapped double stranded DNA is introduced near the active site, pol X prefers an open conformation, regardless of the salt concentration. Finally, under physiological conditions, in the presence of both gapped DNA and correct incoming nucleotide, and two divalent ions, the thumb subdomain of pol X undergoes a large conformational change, closing upon the DNA. These results predict for pol X a substrate-induced conformational change triggered by the presence of DNA and the correct incoming nucleotide in the active site, as in DNA polymerase beta. The simulations also suggest specific experiments (e.g., for mutants Phe-102Ala, Val 120Gly, and Lys-85Val that may reveal crucial DNA binding and active-site organization roles) to further elucidate the fidelity mechanism of pol X. PMID- 16214866 TI - Mechanics and dynamics of actin-driven thin membrane protrusions. AB - Motile cells explore their surrounding milieu by extending thin dynamic protrusions, or filopodia. The growth of filopodia is driven by actin filament bundles that polymerize underneath the cell membrane. We compute the mechanical and dynamical features of the protrusion growth process by explicitly incorporating the flexible plasma membrane. We find that a critical number of filaments are needed to generate net filopodial growth. Without external influences, the filopodium can extend indefinitely up to the buckling length of the F-actin bundle. Dynamical calculations show that the protrusion speed is enhanced by the thermal fluctuations of the membrane; a filament bundle encased in a flexible membrane grows much faster. The protrusion speed depends directly on the number and spatial arrangement of the filaments in the bundle and whether the filaments are tethered to the membrane. Filopodia also attract each other through distortions of the membrane. Spatially close filopodia will merge to form a larger one. Force-velocity relationships mimicking micromanipulation experiments testing our predictions are computed. PMID- 16214867 TI - Is the mechanical activity of epithelial cells controlled by deformations or forces? AB - The traction forces developed by cells depend strongly on the substrate rigidity. In this letter, we characterize quantitatively this effect on MDCK epithelial cells by using a microfabricated force sensor consisting in a high-density array of soft pillars whose stiffness can be tailored by changing their height and radius to obtain a rigidity range from 2 nN/microm up to 130 nN/microm. We find that the forces exerted by the cells are proportional to the spring constant of the pillars meaning that, on average, the cells deform the pillars by the same amount whatever their rigidity. The relevant parameter may thus be a deformation rather than a force. These dynamic observations are correlated with the reinforcement of focal adhesions that increases with the substrate rigidity. PMID- 16214868 TI - Dipolar coupling between nitroxide spin labels: the development and application of a tether-in-a-cone model. AB - A tether-in-a-cone model is developed for the simulation of electron paramagnetic resonance spectra of dipolar coupled nitroxide spin labels attached to tethers statically disordered within cones of variable halfwidth. In this model, the nitroxides adopt a range of interprobe distances and orientations. The aim is to develop tools for determining both the distance distribution and the relative orientation of the labels from experimental spectra. Simulations demonstrate the sensitivity of electron paramagnetic resonance spectra to the orientation of the cones as a function of cone halfwidth and other parameters. For small cone halfwidths (< approximately 40 degrees ), simulated spectra are strongly dependent on the relative orientation of the cones. For larger cone halfwidths, spectra become independent of cone orientation. Tether-in-a-cone model simulations are analyzed using a convolution approach based on Fourier transforms. Spectra obtained by the Fourier convolution method more closely fit the tether-in-a-cone simulations as the halfwidth of the cone increases. The Fourier convolution method gives a reasonable estimate of the correct average distance, though the distance distribution obtained can be significantly distorted. Finally, the tether-in-a-cone model is successfully used to analyze experimental spectra from T4 lysozyme. These results demonstrate the utility of the model and highlight directions for further development. PMID- 16214869 TI - Probing the mechanical folding kinetics of TAR RNA by hopping, force-jump, and force-ramp methods. AB - Mechanical unfolding and refolding of single RNA molecules have previously been observed in optical traps as sudden changes in molecular extension. Two methods have been traditionally used: "force-ramp", with the applied force continuously changing, and "hopping". In hopping experiments the force is held constant and the molecule jumps spontaneously between two different states. Unfolding/refolding rates are measured directly, but only over a very narrow range of forces. We have now developed a force-jump method to measure the unfolding and refolding rates independently over a wider range of forces. In this method, the applied force is rapidly stepped to a new value and either the unfolding or refolding event is monitored through changes in the molecular extension. The force-jump technique is compared to the force-ramp and hopping methods by using a 52-nucleotide RNA hairpin with a three-nucleotide bulge, i.e., the transactivation response region RNA from the human immunodeficiency virus. We find the unfolding kinetics and Gibbs free energies obtained from all three methods to be in good agreement. The transactivation response region RNA hairpin unfolds in an all-or-none two-state reaction at any loading rate with the force ramp method. The unfolding reaction is reversible at small loading rates, but shows hysteresis at higher loading rates. Although the RNA unfolds and refolds without detectable intermediates in constant-force conditions (hopping and force jump), it shows partially folded intermediates in force-ramp experiments at higher unloading rates. Thus, we find that folding of RNA hairpins can be more complex than a simple single-step reaction, and that application of several methods can improve understanding of reaction mechanisms. PMID- 16214870 TI - Organelle transport along microtubules in Xenopus melanophores: evidence for cooperation between multiple motors. AB - Xenopus melanophores have pigment organelles or melanosomes which, in response to hormones, disperse in the cytoplasm or aggregate in the perinuclear region. Melanosomes are transported by microtubule motors, kinesin-2 and cytoplasmic dynein, and an actin motor, myosin-V. We explored the regulation of melanosome transport along microtubules in vivo by using a new fast-tracking routine, which determines the melanosome position every 10 ms with 2-nm precision. The velocity distribution of melanosomes transported by cytoplasmic dynein or kinesin-2 under conditions of aggregation and dispersion presented several peaks and could not be fit with a single Gaussian function. We postulated that the melanosome velocity depends linearly on the number of active motors. According to this model, one to three dynein molecules transport each melanosome in the minus-end direction. The transport in the plus-end direction is mainly driven by one to two copies of kinesin-2. The number of dyneins transporting a melanosome increases during aggregation, whereas the number of active kinesin-2 stays the same during aggregation and dispersion. Thus, the number of active dynein molecules regulates the net direction of melanosome transport. The model also shows that multiple motors of the same polarity cooperate during the melanosome transport, whereas motors of opposite polarity do not compete. PMID- 16214871 TI - Lipid asymmetry in DLPC/DSPC-supported lipid bilayers: a combined AFM and fluorescence microscopy study. AB - A fundamental attribute of cell membranes is transmembrane asymmetry, specifically the formation of ordered phase domains in one leaflet that are compositionally different from the opposing leaflet of the bilayer. Using model membrane systems, many previous studies have demonstrated the formation of ordered phase domains that display complete transmembrane symmetry; but there have been few reports on the more biologically relevant asymmetric membrane structures. Here we report on a combined atomic force microscopy and fluorescence microscopy study whereby we observe three different states of transmembrane symmetry in phase-separated supported lipid bilayers formed by vesicle fusion. We find that if the leaflets differ in gel-phase area fraction, then the smaller domains in one leaflet are in registry with the larger domains in the other leaflet and the system is dynamic. In a presumed lipid flip-flop process similar to Ostwald ripening, the smaller domains in one leaflet erode away whereas the large domains in the other leaflet grow until complete compositional asymmetry is reached and remains stable. We have quantified this evolution and determined that the lipid flip-flop event happens most frequently at the interface between symmetric and asymmetric DSPC domains. If both leaflets have identical area fraction of gel-phase, gel-phase domains are in registry and are static in comparison to the first state. The stability of these three DSPC domain distributions, the degree of registry observed, and the domain immobility have biological significance with regards to maintenance of lipid asymmetry in living cell membranes, communication between inner leaflet and outer leaflet, membrane adhesion, and raft mobility. PMID- 16214872 TI - A stochastic two-dimensional model of intercellular Ca2+ wave spread in glia. AB - We describe a two-dimensional stochastic model of intercellular Ca(2+) wave (ICW) spread in glia that includes contributions of external stimuli, ionotropic and metabotropic P2 receptors, exo- and ecto-nucleotidases, second messengers, and gap junctions. In this model, an initial stimulus evokes ATP and UTP release from a single cell. Agonists diffuse and are degraded both in bulk solution and at cell surfaces. Ca(2+) elevation in individual cells is determined by bound agonist concentrations s and by number and features of P2 receptors summed with that generated by IP(3) diffusing through gap junction channels. Variability of ICWs is provided by randomly distributing a predetermined density of cells in a rectangular grid and by randomly selecting within intervals values characterizing the extracellular compartment, individual cells, and interconnections with neighboring cells. Variability intervals were obtained from experiments on astrocytoma cells transfected to express individual P2 receptors and/or the gap junction protein connexin43. The simulation program (available as Supplementary Material) permits individual alteration of ICW components, allowing comparison of simulations with data from cells expressing connexin43 and/or various P2 receptor subtypes. Such modeling is expected to be useful for testing phenomenological hypotheses and in understanding consequences of alteration of system components under experimental or pathological conditions. PMID- 16214873 TI - Effects of disulfide bonds on folding behavior and mechanism of the beta-sheet protein tendamistat. AB - Tendamistat, a small disulfide-bonded beta-sheet protein, and its three single/double-disulfide mutants are investigated by using a modified Go-like model, aiming to understand the folding mechanism of disulfide-bonded protein as well as the effects of removal of disulfide bond on the folding process. Our simulations show that tendamistat and its two single-disulfide mutants are all two-state folders, consistent with the experimental observations. It is found that the disulfide bonds as well as three hydrogen bonds between the N-terminal loop-0 and strand-6 are of significant importance for the folding of tendamistat. Without these interactions, their two-state behaviors become unstable and the predictions of the model are inconsistent with experiments. In addition, the effect of disulfide bonds on the folding process are studied by comparing the wild-type tendamistat and its two mutants; it is found that the removal of either of the C11-C27 or C45-C73 disulfide bond leads to a large decrease in the thermodynamical stability and loss of structure in the unfolded state, and the effect of the former is stronger than that of the later. These simulation results are in good agreement with experiments and, thus, validate our model. Based on the same model, the detailed folding pathways of the wild-type tendamistat and two mutants are studied, and the effect of disulfide bonds on the folding kinetics are discussed. The obtained results provide a detailed folding picture of these proteins and complement experimental findings. Finally, the folding nuclei predicted to be existent in this protein tendamistat as well as its mutants are firstly identified in this work. The positions of the nucleus are consistent with those argued in experimental studies. Therefore, a nucleation/growth folding mechanism that can explain the two-state folding manner is clearly characterized. Moreover, the effect by the removal of each disulfide bond on the folding thermodynamics and dynamics can also be well interpreted from their influence on the folding nucleus. The implementation of this work indicates that the modified Go-like model really describes the folding behavior of protein tendamistat and could be used to study the folding of other disulfide-bonded proteins. PMID- 16214875 TI - Ca2+ changes the force sensitivity of the hair-cell transduction channel. AB - The mechanically gated transduction channels of vertebrate hair cells tend to close in approximately 1 ms after their activation by hair bundle deflection. This fast adaptation is correlated with a quick negative movement of the bundle (a "twitch"), which can exert force and may mediate an active mechanical amplification of sound stimuli in hearing organs. We used an optical trap to deflect bullfrog hair bundles and to measure bundle movement while controlling Ca(2+) entry with a voltage clamp. The twitch elicited by repolarization of the cell varied with force applied to the bundle, going to zero where channels were all open or closed. The force dependence is quantitatively consistent with a model in which a Ca(2+)-bound channel requires approximately 3 pN more force to open, and rules out other models for the site of Ca(2+) action. In addition, we characterized a faster, voltage-dependent "flick", which requires intact tip links but not current through transduction channels. PMID- 16214874 TI - Three-dimensional visualization of FKBP12.6 binding to an open conformation of cardiac ryanodine receptor. AB - The cardiac isoform of the ryanodine receptor (RyR2) from dog binds predominantly a 12.6-kDa isoform of the FK506-binding protein (FKBP12.6), whereas RyR2 from other species binds both FKBP12.6 and the closely related isoform FKBP12. The role played by FKBP12.6 in modulating calcium release by RyR2 is unclear at present. We have used cryoelectron microscopy and three-dimensional (3D) reconstruction techniques to determine the binding position of FKBP12.6 on the surface of canine RyR2. Buffer conditions that should favor the "open" state of RyR2 were used. Quantitative comparison of 3D reconstructions of RyR2 in the presence and absence of FKBP12.6 reveals that FKBP12.6 binds along the sides of the square-shaped cytoplasmic region of the receptor, adjacent to domain 9, which forms part of the four clamp (corner-forming) structures. The location of the FKBP12.6 binding site on "open" RyR2 appears similar, but slightly displaced (by 1-2 nm) from that found previously for FKBP12 binding to the skeletal muscle ryanodine receptor that was in the buffer that favors the "closed" state. The conformation of RyR2 containing bound FKBP12.6 differs considerably from that depleted of FKBP12.6, particularly in the transmembrane region and in the clamp structures. The x-ray structure of FKBP12.6 was docked into the region of the 3D reconstruction that is attributable to bound FKBP12.6, to show the relative orientations of amino acid residues (Gln-31, Asn-32, Phe-59) that have been implicated as being critical in interactions with RyR2. A thorough understanding of the structural basis of RyR2-FKBP12.6 interaction should aid in understanding the roles that have been proposed for FKBP12.6 in heart failure and in certain forms of sudden cardiac death. PMID- 16214876 TI - Pigment-pigment interactions in PCP of Amphidinium carterae investigated by nonlinear polarization spectroscopy in the frequency domain. AB - Peridinin-chlorophyll a-protein (PCP) is a unique antenna complex in dinoflagellates that employs peridinin (a carotenoid) as its main light harvesting pigment. Strong excitonic interactions between peridinins, as well as between peridinins and chlorophylls (Chls) a, can be expected from the short intermolecular distances revealed by the crystal structure. Different experimental approaches of nonlinear polarization spectroscopy in the frequency domain (NLPF) were used to investigate the various interactions between pigments in PCP of Amphidinium carterae at room temperature. Lineshapes of NLPF spectra indicate strong excitonic interactions between the peridinin's optically allowed S(2) (1Bu(+)) states. A comprehensive subband analysis of the distinct NLPF spectral substructure in the peridinin region allows us to assign peridinin subbands to the two Chls a in PCP having different S(1)-state lifetimes. Peridinin subbands at 487, 501, and 535 nm were assigned to the longer-lived Chl, whereas a peridinin subband peaking at 515 nm was detected in both clusters. Certain peridinin(s), obviously corresponding to the subband centered at 487 nm, show(s) specific (possibly Coulombic?) interaction between the optically dark S(1)(2A(g)(-)) and/or intramolecular charge-transfer (ICT) state and S(1) of Chl a. The NLPF spectrum, hence, indicates that this peridinin state is approximately isoenergetic or slightly above S(1) of Chl a. A global subband analysis of absorption and NLPF spectra reveals that the Chl a Q(y)-band consists of two subbands (peaking at 669 and 675 nm and having different lifetimes), confirmed by NLPF spectra recorded at high pump intensities. At the highest applied pump intensities an additional band centered at S(1)/ICT transition of peridinin. PMID- 16214877 TI - Comparison of galantamine and donepezil for effects on nerve growth factor, cholinergic markers, and memory performance in aged rats. AB - This study was designed to determine 1) whether repeated exposures to the acetylcholinesterase inhibitors (AChEIs) galantamine (GAL) or donepezil (DON) resulted in positive effects on nerve growth factor (NGF) and its receptors, cholinergic proteins, and cognitive function in the aged rat, and 2) whether GAL had any advantages over DON given its allosteric potentiating ligand (APL) activity at nicotinic acetylcholine receptors. Behavioral tests (i.e., water maze and light/dark box) were conducted in aged Fisher 344 rats during 15 days of repeated (subcutaneous) exposure to either GAL (3.0 or 6.0 mg/kg/day) or DON (0.375 or 0.75 mg/kg/day). Forty-eight hours after the last drug injection, cholinergic receptors were measured by [(125)I]-(+/-)-exo-2-(2-iodo-5-pyridyl)-7 azabicyclo[2.2.1]heptane ([(125)I]IPH; epibatidine analog), (125)I-alpha bungarotoxin ((125)I-BTX), [(3)H]pirenzepine ([(3)H]PRZ), and [(3)H]-5,11-dihydro 11-[((2-(2-((dipropylamino)methyl)-1-piperidinyl)ethyl)amino)carbonyl]-6H pyrido(2,3-b)(1,4)-benzodiazepin-6-one methanesulfonate ([(3)H]AFDX-384, or [(3)H]AFX) autoradiography. Immunochemical methods were used to measure NGF, high (TrkA and phospho-TrkA)- and low (p75 neurotrophin receptor)-affinity NGF receptors, choline acetyltransferase (ChAT), and the vesicular acetylcholine transporter (VAChT) in memory-related brain regions. Depending on dose, both GAL and DON enhanced spatial learning (without affecting anxiety levels) and increased [(125)I]IPH, [(3)H]PRZ, and [(3)H]AFX (but decreased (125)I-BTX) binding in some cortical and hippocampal brain regions. Neither AChEI was associated with marked changes in NGF, NGF receptors, or VAChT, although DON did moderately increase ChAT in the basal forebrain and hippocampus. The results suggest that repeated exposures to either GAL or DON results in positive (and sustained) behavioral and cholinergic effects in the aged mammalian brain but that the APL activity of GAL may not afford any advantage over acetylcholinesterase inhibition alone. PMID- 16214878 TI - Repression of cytochrome P450 activity in human hepatocytes in vitro by a novel hepatotrophic factor, augmenter of liver regeneration. AB - Pathological disorders of the liver were shown to be associated with an impairment of hepatic drug metabolism mediated in part by growth factors. Augmenter of liver regeneration (ALR) is a novel liver-specific hepatotrophic growth factor, whereas its action on cytochrome P450 (P450) metabolism is completely unknown. Application of ALR to primary human hepatocytes in vitro reduced P450 isoenzyme activities (1A2 and 2A6) in a dose-dependent manner. Time course analysis revealed that the maximal inhibitory effect was reached after 24 to 72 h of exposure with 50 nM ALR. The reduction of basal activities upon ALR treatment was 35% for CYP1A2, 56% for CYP2A6, 18% for CYP2B6, and 45% for CYP2E1. Additionally, after induction of P450 with specific inducers, ALR revealed an inhibitory effect on the isoenzyme activities (CYP1A2, 41%; CYP2B6, 35%). Investigations of protein and mRNA expression of basal and induced CYP1A2 and CYP3A4 after ALR treatment by Western blotting and real-time reverse transcriptase-polymerase chain reaction, respectively, suggest a regulation on the transcriptional level. Furthermore, ALR treatment increased nuclear factor kB activity and reduced constitutive androstane receptor but not pregnane X receptor or aryl hydrocarbon receptor expression. In contrast, ALR revealed no effects on phase II reactions (glutathione/oxidized glutathione, UDP-glucuronyltransferase conjugation). Our results indicate that ALR, as a member of hepatotrophic factors, down-regulates basal and induced P450 in human liver and therefore cross links growth signals to regulation of hepatic metabolism. These findings further imply a possible role of ALR in drug interactions during impaired hepatic function, whereas liver regeneration is triggered. PMID- 16214879 TI - Dominant role for calpain in thromboxane-induced neuromicrovascular endothelial cytotoxicity. AB - Thromboxane A(2) (TXA(2)) is an important lipid mediator generated during oxidative stress and implicated in ischemic neural injury. This autacoid was recently shown to partake in this injury process by directly inducing endothelial cytotoxicity. We explored the mechanisms for this TXA(2)-evoked neural microvascular endothelial cell death. Stable TXA(2) mimetics 5-heptenoic acid, 7 [6-(3-hydroxy-1-octenyl)-2-oxabicyclo[2.2.1]hept-5-yl]-[1R [1alpha,4alpha,5beta(Z),6alpha,(1E,3S)]]-9,11-dedioxy-9alpha,11alpha-methanolpoxy (U-46619) [as well as [1S-[1alpha,2alpha(Z),3beta(1E,3S(*)),4alpha]]-7-[3-[3 hydroxy-4-(4-iodophenoxy)-1-butenyl]-7-oxabicyclo[2.1.1]-hept-2-yl]-5-heptenoic acid; I-BOP] induced a retinal microvascular degeneration in rat pups in vivo and in porcine retinal explants ex vivo and death of porcine brain endothelial cells (in culture). TXA(2) dependence of these effects was corroborated by antagonism using the selective TXA(2) receptor blocker (-)-6,8-difluoro-9-p-methyl-sulfonyl benzyl-1,2,3,4-tetrahydrocarbazol-1-yl-acetic acid (L670596). In all cases, neurovascular endothelial cell death was prevented by pan-calpain and specific m calpain inhibitors but not by caspase-3 or pan-caspase inhibitors. Correspondingly, TXA(2) (mimetics) augmented generation of known active m-calpain (but not mu-calpain) form and increased the activity of m-calpain (cleavage of fluorogenic substrate N-succinyl-Leu-Leu-Val-Tyr-7-amino-4-methylcoumarin; and of alpha-spectrin into specific fragments) but not of pan-caspase or specific caspase-3 (respectively, using sulforhodamine-Val-Arg-Asp-fluoromethyl ketone and detecting its active 17- and 12-kDa fragments). Interestingly, these effects were phospholipase C (PLC)-dependent [associated with increase in inositol triphosphate and inhibited by PLC blocker 1-[6-[[17beta-methoxyestra-1,3,5(10) trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U73122)] and required calcium but were not associated with increased intracellular calcium. U-46619-induced calpain activation resulted in translocation of Bax to the mitochondria, loss of polarization of the latter (using potentiometric probe 5,5',6,6'-tetrachloro 1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide; JC-1) and in turn release of cytochrome c into the cytosol and depletion of cellular ATP; these effects were all blocked by calpain inhibitors. Overall, this work identifies (specifically) m-calpain as a dominant protease in TXA(2)-induced neurovascular endothelial cell death. PMID- 16214881 TI - Process of care and mortality of stroke patients with and without a do not resuscitate order in the West Midlands, UK. AB - OBJECTIVE: . To compare the process of care of stroke patients with and without a do not resuscitate (DNR) order. DESIGN: Retrospective case note review with prospective follow up of mortality. SETTING: Seven acute hospitals, with stroke units, in the West Midlands, UK. PARTICIPANTS: A random sample of patients (n = 702) admitted to hospital with acute stroke over a twelve month period. MAIN OUTCOME MEASURES: Case mix and process of care measures derived from the intercollegiate stroke audit package. Thirty day and one year mortality. RESULTS: About one-third (34%, 238/702) of stroke patients had DNR orders. The thirty-day mortality for DNR patients was 67% (160/238) versus 10% (46/449) for patients without DNR orders. DNR patients had significantly worse case-mix profile than non-DNR patients - median age 81 y vs 75y; fully conscious 36% vs 79%, able to walk 1% vs 21% and no loss of power in either arm 5% vs 24% (all p < 0.0001). DNR patients were more likely to be assessed early by a speech and language therapist (77% vs 59%, p < 0.001), but less likely to receive the majority of their care in a stroke/rehabilitation unit (20% vs 57%, p < 0.0001), or be cared for on a stroke unit or by a stroke team (42% vs 70%, p <0.0001), or had a description of the site of the cerebral lesion (31% vs 38%, p = 0.05) or be given aspirin (30% vs 42%, p = 0.007). CONCLUSIONS: Stroke patients with a DNR order are not receiving optimum care in that they are not being cared for on stroke units or by specialist teams. This may reflect the inadequate provision of specialist stroke services in the UK. PMID- 16214880 TI - Effects of exogenous and endogenous cannabinoids on GABAergic neurotransmission between the caudate-putamen and the globus pallidus in the mouse. AB - Globus pallidus neurons receive GABAergic input from the caudate-putamen via the striatopallidal pathway. Anatomical studies indicate that many CB(1) cannabinoid receptors are localized on terminals of striatopallidal axons. Accordingly, the hypothesis of the present work was that activation of CB(1) receptors presynaptically inhibits neurotransmission between striatopallidal axons and globus pallidus neurons. In sagittal mouse brain slices, striatopallidal axons were electrically stimulated in the caudate-putamen, and the resulting GABAergic inhibitory postsynaptic currents (IPSCs) were recorded in globus pallidus neurons. The synthetic cannabinoid receptor agonists R(+)-[2,3-dihydro-5-methyl-3 [(morpholinyl)methyl] pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-naphthalenyl) methanone mesylate (WIN55212-2) and (-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl) phenyl]-trans-4-(3-hydroxy-propyl)-cyclohexanol (CP55940) decreased the amplitude of IPSCs. The CB(1) receptor antagonist rimonabant prevented the inhibition by WIN55212-2, pointing to involvement of CB(1) receptors. Depolarization of globus pallidus neurons induced a weak and short-lasting suppression of IPSCs [i.e., depolarization-induced suppression of inhibition (DSI) occurred]. Prevention of DSI by rimonabant indicates that endocannabinoids released from the postsynaptic neurons acted on CB(1) receptors to suppress synaptic transmission. WIN55212-2 did not modify currents in globus pallidus neurons elicited by GABA released from its chemically bound ("caged") form by a flash pulse, suggesting that WIN55212-2 depressed neurotransmission presynaptically. For studying the mechanism of the inhibition of GABA release, terminals of striatopallidal axons were labeled with a calcium-sensitive fluorescent dye. WIN55212-2 depressed the action potential evoked increase in axon terminal calcium concentration. The results show that activation of CB(1) receptors by exogenous and endogenous cannabinoids leads to presynaptic inhibition of neurotransmission between striatopallidal axons and globus pallidus neurons. Depression of the action potential-evoked calcium influx into axon terminals is the probable mechanism of this inhibition. PMID- 16214882 TI - Use of risk-adjusted change in health status to assess the performance of integrated service networks in the Veterans Health Administration. AB - OBJECTIVE: Health outcome assessments have become an expectation of regulatory and accreditation agencies. We examined whether a clinically credible risk adjustment methodology for the outcome of change in health status can be developed for performance assessment of integrated service networks. STUDY DESIGN: Longitudinal study. SETTING: Outpatient. STUDY PARTICIPANTS: Thirty-one thousand eight hundred and twenty-three patients from 22 Veterans Health Administration (VHA) integrated service networks were followed for 18 months. MAIN MEASURE: The physical (PCS) and mental (MCS) component scales from the Veterans Rand 36-items Health Survey (VR-36) and mortality. The outcomes were decline in PCS (decline in PCS scores greater than -6.5 points or death) and MCS (decline in MCS scores greater than -7.9 points). RESULTS: Four thousand three hundred and twenty-eight (13.6%) patients showed a decline in PCS scores greater than -6.5 points, 4322 (13.5%) had a decline in MCS scores by more than -7.9 points, and 1737 died (5.5%). Multivariate logistic regression models were used to adjust for case-mix. The models performed reasonably well in cross-validated tests of discrimination (c-statistics = 0.72 and 0.68 for decline in PCS and MCS, respectively) and calibration. The resulting risk-adjusted rates of decline in PCS and MCS and ranks of the networks differed considerably from unadjusted ratings. CONCLUSION: It is feasible to develop clinically credible risk adjustment models for the outcomes of decline in PCS and MCS. Without adequate controls for case-mix, we could not determine whether poor patient outcomes reflect poor performance, sicker patients, or other factors. This methodology can help to measure and report the performance of health care systems. PMID- 16214883 TI - Diagnostic imaging pathways: development, dissemination, implementation, and evaluation. AB - ISSUE: There are signs of inappropriate application of medical imaging to diagnosis. Inappropriate imaging is a threat to effective diagnosis and effective allocation of resources. The development and deployment of knowledge-based clinical decision support systems is one strategy to reduce inappropriate imaging. DEVELOPMENT OF IMAGING PATHWAYS: A suite of 78 imaging pathways was conceived both as a decision support and educational tool. The pathways were drafted by imaging specialists, but further developed and modified, based on graded evidence and input from requesting clinicians. An electronic environment was developed to contain and deliver the pathways. DISSEMINATION AND IMPLEMENTATION: Imaging pathways were distributed via a hospital local area network and on compact disk. A multifaceted approach was used to raise general awareness of the pathways, followed by intensive 'marketing' activities. Two groups of clinicians were targeted; hospital-based clinicians and general practitioners. EVALUATION: There was increased awareness of imaging pathways. Clinicians judged them to be useful for education and decision support. The method of electronic delivery was adequate. Knowledge of diagnostic imaging and requesting behaviour tended to become more aligned with the pathways. The central objective to reduce inappropriate medical imaging seems to be achievable. LESSONS LEARNED: There is scope to improve the content and the electronic environment, achieve better integration into decision-making processes, and achieve better compliance. A linkage between imaging pathways and electronic requesting could provide alerts to 'non-compliant' requesting. The assignment of a higher cost, or a lower remuneration, to non-authorized and non-compliant imaging would provide tangible incentive to comply, unless there are compelling clinical contraindications. PMID- 16214884 TI - Microfluidic gas-flow profiling using remote-detection NMR. AB - We have used nuclear magnetic resonance (NMR) to obtain spatially and temporally resolved profiles of gas flow in microfluidic devices. Remote detection of the NMR signal both overcomes the sensitivity limitation of NMR and enables time-of flight measurement in addition to spatially resolved imaging. Thus, detailed insight is gained into the effects of flow, diffusion, and mixing in specific geometries. The ability for noninvasive measurement of microfluidic flow, without the introduction of foreign tracer particles, is unique to this approach and is important for the design and operation of microfluidic devices. Although here we demonstrate an application to gas flow, extension to liquids, which have higher density, is implicit. PMID- 16214885 TI - Blocking anthrax lethal toxin at the protective antigen channel by using structure-inspired drug design. AB - Bacillus anthracis secretes three polypeptides: protective antigen (PA), lethal factor (LF), and edema factor (EF), which interact at the surface of mammalian cells to form toxic complexes. LF and EF are enzymes that target substrates within the cytosol; PA provides a heptameric pore to facilitate LF and EF transport into the cytosol. Other than administration of antibiotics shortly after exposure, there is currently no approved effective treatment for inhalational anthrax. Here we demonstrate an approach to disabling the toxin: high-affinity blockage of the PA pore by a rationally designed low-molecular weight compound that prevents LF and EF entry into cells. Guided by the sevenfold symmetry and predominantly negative charge of the PA pore, we synthesized small cyclic molecules of sevenfold symmetry, beta-cyclodextrins chemically modified to add seven positive charges. By channel reconstitution and high-resolution conductance recording, we show that per-6-(3-aminopropylthio)-beta-cyclodextrin interacts strongly with the PA pore lumen, blocking PA-induced transport at subnanomolar concentrations (in 0.1 M KCl). The compound protected RAW 264.7 mouse macrophages from cytotoxicity of anthrax lethal toxin (= PA + LF). More importantly, it completely protected the highly susceptible Fischer F344 rats from lethal toxin. We anticipate that this approach will serve as the basis for a structure-directed drug discovery program to find new and effective treatments for anthrax. PMID- 16214886 TI - p600, a unique protein required for membrane morphogenesis and cell survival. AB - In this article, we identify and characterize p600, a unique 600-kDa retinoblastoma protein- and calmodulin-binding protein. In the nucleus, p600 and retinoblastoma protein seem to act as a chromatin scaffold. In the cytoplasm, p600 and clathrin form a meshwork structure, which could contribute to cytoskeletal organization and membrane morphogenesis. Reduced expression of p600 with interference RNA abrogates integrin-mediated ruffled membrane formation and, furthermore, prevents activation of integrin-mediated survival pathways. Consequently, knockdown of p600 sensitizes cells to apoptosis induced by cell detachment. These findings provide mechanistic insight into the regulation of membrane-proximal events in tumorigenesis. PMID- 16214887 TI - Predicting the energetics of osmolyte-induced protein folding/unfolding. AB - A primary thermodynamic goal in protein biochemistry is to attain predictive understanding of the detailed energetic changes that are responsible for folding/unfolding. Through use of recently determined free energies of side-chain and backbone transfer from water to osmolytes and Tanford's transfer model, we demonstrate that the long-sought goal of predicting solvent-dependent cooperative protein folding/unfolding free-energy changes (m values) can be achieved. Moreover, the approach permits dissection of the folding/unfolding free-energy changes into individual contributions from the peptide backbone and residue side chains. PMID- 16214888 TI - Crystal structure of the archaeal ammonium transporter Amt-1 from Archaeoglobus fulgidus. AB - Ammonium transporters (Amts) are integral membrane proteins found in all kingdoms of life that fulfill an essential function in the uptake of reduced nitrogen for biosynthetic purposes. Amt-1 is one of three Amts encoded in the genome of the hyperthermophilic archaeon Archaeoglobus fulgidus. The crystal structure of Amt-1 shows a compact trimer with 11 transmembrane helices per monomer and a central channel for substrate conduction in each monomer, similar to the known crystal structure of AmtB from Escherichia coli. Xenon derivatization has been used to identify apolar regions of Amt-1, emphasizing not only the hydrophobicity of the substrate channel but also the unexpected presence of extensive internal cavities that should be detrimental for protein stability. The substrates ammonium and methylammonium have been used for cocrystallization experiments with Amt-1, but the identification of binding sites that are distinct from water positions is not unambiguous. The well ordered cytoplasmic C terminus of the protein in the Amt-1 structure has allowed for the construction of a docking model between Amt-1 and a homology model for its physiological interaction partner, the P(II) protein GlnB 1. In this model, GlnB-1 binds tightly to the cytoplasmic face of the transporter, effectively blocking conduction through the three individual substrate channels. PMID- 16214889 TI - The UGT73C5 of Arabidopsis thaliana glucosylates brassinosteroids. AB - Steroid hormones are essential for development, and the precise control of their homeostasis is a prerequisite for normal growth. UDP-glycosyltransferases (UGTs) are considered to play an important regulatory role in the activity of steroids in mammals and insects. This study provides an indication that a UGT accepting plant steroids as substrates functions in brassinosteroid (BR) homeostasis. The UGT73C5 of Arabidopsis thaliana catalyses 23-O-glucosylation of the BRs brassinolide (BL) and castasterone. Transgenic plants overexpressing UGT73C5 displayed BR-deficient phenotypes and contained reduced amounts of BRs. The phenotype, which was already apparent in seedlings, could be rescued by application of BR. In feeding experiments with BL, wild-type seedlings converted BL to the 23-O-glucoside; in the transgenic lines silenced in UGT73C5 expression, no 23-O-glucoside was detected, implying that this UGT is the only enzyme that catalyzes BL-23-O-glucosylation in seedlings. Plant lines in which UGT73C5 expression was altered also displayed hypocotyl phenotypes previously described for seedlings in which BR inactivation by hydroxylation was changed. These data support the hypothesis that 23-O-glucosylation of BL is a function of UGT73C5 in planta, and that glucosylation regulates BR activity. PMID- 16214891 TI - Transposon-mediated insertional mutagenesis revealed the functions of animal cellulose synthase in the ascidian Ciona intestinalis. AB - Tunicates are the only animals that perform cellulose biosynthesis. The tunicate gene for cellulose synthase, Ci-CesA, was likely acquired by horizontal transfer from bacteria and was a key innovation in the evolution of tunicates. Transposon based mutagenesis in an ascidian, Ciona intestinalis, has generated a mutant, swimming juvenile (sj). Ci-CesA is the gene responsible for the sj mutant, in which a drastic reduction in cellulose was observed in the tunic. Furthermore, during metamorphosis, which in ascidians convert the vertebrate-like larva into a sessile filter feeder, sj showed abnormalities in the order of metamorphic events. In normal larvae, the metamorphic events in the trunk region are initiated after tail resorption. In contrast, sj mutant larvae initiated the metamorphic events in the trunk without tail resorption. Thus, sj larvae show a "swimming juvenile" phenotype, the juvenile-like trunk structure with a complete tail and the ability to swim. It is likely that ascidian cellulose synthase is required for the coordination of the metamorphic events in the trunk and tail in addition to cellulose biosynthesis. PMID- 16214890 TI - Rab11a and myosin Vb are required for bile canalicular formation in WIF-B9 cells. AB - Hepatocytes polarize by forming functionally distinct sinusoidal (basolateral) and canalicular (apical) plasma membrane domains. Two distinct routes are used for delivery of membrane proteins to the canaliculus. Proteins having glycosylphosphatidylinositol anchors or single transmembrane domains are targeted to the sinusoidal plasma membrane from where they transcytose to the canalicular domain. In contrast, apical ATP-binding-cassette (ABC) transporters, which are required for energy-dependent biliary secretion of bile acids (ABCB11), phospholipids (ABCB4), and nonbile acid organic anions (ABCC2), lack initial residence in the basolateral plasma membrane and traffic directly from Golgi membranes to the canalicular membrane. While investigating mechanisms of apical targeting in WIF-B9 cells, a polarized hepatic epithelial cell line, we observed that rab11a is required for canalicular formation. Knockdown of rab11a or overexpression of the rab11a-GDP locked form prevented canalicular formation as did overexpression of the myosin Vb motorless tail domain. In WIF-B9 cells, which lack bile canaliculi, apical ABC transporters colocalized with transcytotic membrane proteins in rab11a-containing endosomes and, unlike the transcytotic markers, did not distribute to the plasma membrane. We propose that polarization of hepatocytes (i.e., canalicular biogenesis) requires recruitment of rab11a and myosin Vb to intracellular membranes that contain apical ABC transporters and transcytotic markers, permitting their targeting to the plasma membrane. In this model, polarization is initiated upon delivery of rab11a-myosin Vb-containing membranes to the surface, which causes plasma membrane at the site of delivery to differentiate into apical domain (bile canaliculus). PMID- 16214892 TI - Signal propagation from membrane messengers to nuclear effectors revealed by reporters of phosphoinositide dynamics and Akt activity. AB - Among various second messengers, phosphatidylinositol 3,4,5-triphosphate (PIP3) and phosphatidylinositol 3,4-bisphosphate [PI(3,4)P2] regulate a variety of cellular processes, such as cell survival, polarization, and proliferation. Many of these functions are achieved via activation of serine/threonine kinase Akt. To investigate the spatiotemporal regulation of these lipids, we constructed a genetically targetable phosphoinositide (PI) indicator by sandwiching pleckstrin homology (PH) domain of Akt and a "pseudoligand" containing acidic amino acid residues, between cyan and yellow mutants of GFP. In living cells, elevations in PIP3 and PI(3,4)P2 by growth factor-induced activation of phosphatidylinositol 3 kinase (PI3K) resulted in a change in fluorescence resonance energy transfer (FRET) between the fluorescent proteins, increasing yellow to cyan emission ratios by 10-30%. This response can be reversed by inhibiting PI3K and abolished by mutating the critical residues responsible for PI binding. Differential dynamics of PIs were observed at plasma membrane of NIH 3T3 cells, stimulated by various growth factors. On the other hand, the nuclear targeted indicator showed no response within an hour after platelet-derived growth factor stimulation, suggesting that no appreciable amounts of accessible PIP3 and PI(3,4)P2 were produced in the nucleus. Furthermore, simultaneous imaging of a plasma membrane targeted PI indicator and a nuclear-targeted Akt activity reporter revealed a gradual and sustained accumulation of Akt activity in the nucleus after rapid and transient production of PIP3 and PI(3,4)P2 at plasma membrane in the same cell. Thus, signal propagation from the lipid messengers at plasma membrane to the effectors in the nucleus is precisely controlled by kinases as well as lipid and protein phosphatases. PMID- 16214893 TI - The Ca2+-induced methyltransferase xPRMT1b controls neural fate in amphibian embryo. AB - We have previously shown that an increase in intracellular Ca2+ is both necessary and sufficient to commit ectoderm to a neural fate in Xenopus embryos. However, the relationship between this Ca2+ increase and the expression of early neural genes has yet to be defined. Using a subtractive cDNA library between untreated and caffeine-treated animal caps, i.e., control ectoderm and ectoderm induced toward a neural fate by a release of Ca2+, we have isolated the arginine N methyltransferase, xPRMT1b, a Ca2+-induced target gene, which plays a pivotal role in this process. First, we show in embryo and in animal cap that xPRMT1b expression is Ca2+-regulated. Second, overexpression of xPRMT1b induces the expression of early neural genes such as Zic3. Finally, in the whole embryo, antisense approach with morpholino oligonucleotide against xPRMT1b impairs neural development and in animal caps blocks the expression of neural markers induced by a release of internal Ca2+. Our results implicate an instructive role of an enzyme, an arginine methyltransferase protein, in the embryonic choice of determination between epidermal and neural fate. The results presented provide insights by which a Ca2+ increase induces neural fate. PMID- 16214894 TI - Massive sequence perturbation of a small protein. AB - Most protein topologies rarely occur in nature, thus limiting our ability to extract sequence information that could be used to predict structure, function, and evolutionary constraints on protein folds. In principle, the sequence diversity explored by a given protein topology could be expanded by introducing sequence perturbations and selecting variant proteins that fold correctly. However, our capacity to explore sequence space is intrinsically limited by the enormous number of sequences generated from the 20 amino acids and the limited number of variants likely to fold. Here we sought to test whether the sequence space for naturally existing proteins can be explored by simple, sequential degeneration of a complete set of short sequence segments of a model protein, without long-range covariation. Using the Raf ras binding domain as a model of a small protein capable of autonomous folding, we degenerated 72 of 76 positions of the primary structure for the 20 amino acids in segments of four to seven residues defined by secondary structure and selected the folded species for interaction with h-ras by using an in vivo survival-selection assay. The methodology presented allowed for rigorous statistical analysis and comparison of sequence diversity. The ensemble of sequence variants of Raf ras binding domain obtained have recaptured the diversity observed for the ubiquitin-roll topology. A signature sequence for this fold and the implication of this strategy to protein design and structure prediction are discussed. PMID- 16214895 TI - Genetic evidence for a mammalian retromer complex containing sorting nexins 1 and 2. AB - We have previously shown that the putative mammalian retromer components sorting nexins 1 and 2 (Snx1 and Snx2) result in embryonic lethality when simultaneously targeted for deletion in mice, whereas others have shown that Hbeta58 (also known as mVps26), another retromer component, results in similar lethality when targeted for deletion. In the current study, we address the genetic interaction of these mammalian retromer components in mice. Our findings reveal a functional interaction between Hbeta58, SNX1, and SNX2 and strongly suggest that SNX2 plays a more critical role than SNX1 in retromer activity during embryonic development. This genetic evidence supports the existence of mammalian retromer complexes containing SNX1 and SNX2 and identifies SNX2 as an important mediator of retromer biology. Moreover, we find that mammalian retromer complexes containing SNX1 and SNX2 have an essential role in embryonic development that is independent of cation-independent mannose 6-phosphate receptor trafficking. PMID- 16214896 TI - A negative elongation factor for human RNA polymerase II inhibits the anti-arrest transcript-cleavage factor TFIIS. AB - Formation of productive transcription complexes after promoter escape by RNA polymerase II is a major event in eukaryotic gene regulation. Both negative and positive factors control this step. The principal negative elongation factor (NELF) contains four polypeptides and requires for activity the two-polypeptide 5,6-dichloro-1-beta-D-ribobenzimidazole-sensitivity inducing factor (DSIF). DSIF/NELF inhibits early transcript elongation until it is counteracted by the positive elongation factor P-TEFb. We report a previously undescribed activity of DSIF/NELF, namely inhibition of the transcript cleavage factor TFIIS. These two activities of DSIF/NELF appear to be mechanistically distinct. Inhibition of nucleotide addition requires > or = 18 nt of nascent RNA, whereas inhibition of TFIIS occurs at all transcript lengths. Because TFIIS promotes escape from promoter-proximal pauses by stimulating cleavage of back-tracked nascent RNA, TFIIS inhibition may help DSIF/NELF negatively regulate productive transcription. PMID- 16214897 TI - Ectopic DICER-LIKE1 expression in P1/HC-Pro Arabidopsis rescues phenotypic anomalies but not defects in microRNA and silencing pathways. AB - Expression of the viral silencing suppressor P1/HC-Pro in plants causes severe developmental anomalies accompanied by defects in both short interfering RNA (siRNA) and microRNA (miRNA) pathways. P1/HC-Pro transgenic lines fail to accumulate the siRNAs that mediate RNA silencing and are impaired in both miRNA processing and function, accumulating abnormally high levels of miRNA/miRNA* processing intermediates as well as miRNA target messages. Both miRNA and RNA silencing pathways require participation of DICER-LIKE (DCL) ribonuclease III like enzymes. Here, we investigate the effects of overexpressing DCL1, one of four Dicers in Arabidopsis thaliana, on P1/HC-Pro-induced defects in development and small RNA metabolism. Expression of a DCL1 cDNA transgene (35S:DCL1) produced a mild gain-of-function phenotype and largely rescued dcl1 mutant phenotypes. The 35S:DCL1 plants were competent for virus-induced RNA silencing but were impaired in transgene-induced RNA silencing and in the accumulation of some miRNAs. Ectopic DCL1 largely alleviated developmental anomalies in P1/HC-Pro plants but did not correct the P1/HC-Pro-associated defects in small RNA pathways. The ability of P1/HC-Pro plants to suppress RNA silencing and the levels of miRNAs, miRNA*s, and miRNA target messages in these plants were essentially unaffected by ectopic DCL1. These data suggest that P1/HC-Pro defects in development do not result from general impairments in small RNA pathways and raise the possibility that DCL1 participates in processes in addition to miRNA biogenesis. PMID- 16214898 TI - The NAC transcription factors NST1 and NST2 of Arabidopsis regulate secondary wall thickenings and are required for anther dehiscence. AB - In plants, secondary wall thickenings play important roles in various biological processes, although the factors regulating these processes remain to be characterized. We show that expression of chimeric repressors derived from NAC SECONDARY WALL THICKENING PROMOTING FACTOR1 (NST1) and NST2 in Arabidopsis thaliana resulted in an anther dehiscence defect due to loss of secondary wall thickening in anther endothecium. Plants with double, but not single, T-DNA tagged lines for NST1 and NST2 had the same anther-indehiscent phenotype as transgenic plants that expressed the individual chimeric repressors, indicating that NST1 and NST2 are redundant in regulating secondary wall thickening in anther walls. The activity of the NST2 promoter was particularly strong in anther tissue, while that of the NST1 promoter was detected in various tissues in which lignified secondary walls develop. Ectopic expression of NST1 or NST2 induced ectopic thickening of secondary walls in various aboveground tissues. Epidermal cells with ectopic thickening of secondary walls had structural features similar to those of tracheary elements. However, among genes involved in the differentiation of tracheary elements, only those related to secondary wall synthesis were clearly upregulated. None of the genes involved in programmed cell death were similarly affected. Our results suggest NAC transcription factors as possible regulators of secondary wall thickening in various tissues. PMID- 16214900 TI - Crystal structures of a multifunctional triterpene/flavonoid glycosyltransferase from Medicago truncatula. AB - Glycosylation is a ubiquitous reaction controlling the bioactivity and storage of plant natural products. Glycosylation of small molecules is catalyzed by a superfamily of glycosyltransferases (GTs) in most plant species studied to date. We present crystal structures of the UDP flavonoid/triterpene GT UGT71G1 from Medicago truncatula bound to UDP or UDP-glucose. The structures reveal the key residues involved in the recognition of donor substrate and, by comparison with other GT structures, suggest His-22 as the catalytic base and Asp-121 as a key residue that may assist deprotonation of the acceptor by forming an electron transfer chain with the catalytic base. Mutagenesis confirmed the roles of these key residues in donor substrate binding and enzyme activity. Our results provide an initial structural basis for understanding the complex substrate specificity and regiospecificity underlying the glycosylation of plant natural products and other small molecules. This information will direct future attempts to engineer bioactive compounds in crop plants to improve plant, animal, and human health and to facilitate the rational design of GTs to improve the storage and stability of novel engineered bioactive compounds. PMID- 16214901 TI - Structure of Ptr ToxA: an RGD-containing host-selective toxin from Pyrenophora tritici-repentis. AB - Tan spot of wheat (Triticum aestivum), caused by the fungus Pyrenophora tritici repentis, has significant agricultural and economic impact. Ptr ToxA (ToxA), the first discovered proteinaceous host-selective toxin, is produced by certain P. tritici-repentis races and is necessary and sufficient to cause cell death in sensitive wheat cultivars. We present here the high-resolution crystal structure of ToxA in two different crystal forms, providing four independent views of the protein. ToxA adopts a single-domain, beta-sandwich fold of novel topology. Mapping of the existing mutation data onto the structure supports the hypothesized importance of an Arg-Gly-Asp (RGD) and surrounding sequence. Its occurrence in a single, solvent-exposed loop in the protein suggests that it is directly involved in recognition events required for ToxA action. Furthermore, the ToxA structure reveals a surprising similarity with the classic mammalian RGD containing domain, the fibronectin type III (FnIII) domain: the two topologies are related by circular permutation. The similar topologies and the positional conservation of the RGD-containing loop raises the possibility that ToxA is distantly related to mammalian FnIII proteins and that to gain entry it binds to an integrin-like receptor in the plant host. PMID- 16214899 TI - The Arabidopsis cold-responsive transcriptome and its regulation by ICE1. AB - To understand the gene network controlling tolerance to cold stress, we performed an Arabidopsis thaliana genome transcript expression profile using Affymetrix GeneChips that contain approximately 24,000 genes. We statistically determined 939 cold-regulated genes with 655 upregulated and 284 downregulated. A large number of early cold-responsive genes encode transcription factors that likely control late-responsive genes, suggesting a multitude of transcriptional cascades. In addition, many genes involved in chromatin level and posttranscriptional regulation were also cold regulated, suggesting their involvement in cold-responsive gene regulation. A number of genes important for the biosynthesis or signaling of plant hormones, such as abscisic acid, gibberellic acid, and auxin, are regulated by cold stress, which is of potential importance in coordinating cold tolerance with growth and development. We compared the cold-responsive transcriptomes of the wild type and inducer of CBF expression 1 (ice1), a mutant defective in an upstream transcription factor required for chilling and freezing tolerance. The transcript levels of many cold responsive genes were altered in the ice1 mutant not only during cold stress but also before cold treatments. Our study provides a global picture of the Arabidopsis cold-responsive transcriptome and its control by ICE1 and will be valuable for understanding gene regulation under cold stress and the molecular mechanisms of cold tolerance. PMID- 16214902 TI - Diacidic motifs influence the export of transmembrane proteins from the endoplasmic reticulum in plant cells. AB - In yeast and mammals, amino acid motifs in the cytosolic tails of transmembrane domains play a role in protein trafficking by facilitating export from the endoplasmic reticulum (ER). However, little is known about ER export signals of membrane proteins in plants. Therefore, we investigated the role of diacidic motifs in the ER export of Golgi-localized membrane proteins. We show that diacidic motifs perform a significant function in the export of transmembrane proteins to the Golgi apparatus, as mutations of these signals impede the efficient anterograde transport of multispanning, type II, and type I proteins. Furthermore, we demonstrate that diacidic motifs instigate the export of proteins that reside in the ER due to the lengths of their transmembrane domains. However, not all of the diacidic motifs in the cytosolic tails of the proteins studied were equally important in ER export. Transport of Golgi proteins was disrupted only by mutagenesis of specific diacidic signals, suggesting that the protein environment of these signals affects their function. Our findings indicate that diacidic ER export motifs are present and functional in plant membrane proteins and that they are dominant over transmembrane domain length in determining the export of proteins from the ER in plant cells. PMID- 16214904 TI - Long-acting beta2-agonists in asthma: safety concerns. PMID- 16214905 TI - Cancer proteomics: in pursuit of "true" biomarker discovery. PMID- 16214906 TI - Smoking, the missing drug interaction in clinical trials: ignoring the obvious. AB - Tobacco use is universally recognized as the foremost preventable cause of cancer in the United States and globally and is responsible for 30% of all cancer related deaths in the United States. Tobacco use, including exposure to secondhand smoke has been implicated as a causal or contributory agent in an ever expanding list of cancers, including lung, oral cavity and pharynx, pancreas, liver, kidney, ureter, urinary bladder, uterine cervix, and myeloid leukemia. In addition to and independent of the etiologic effects of tobacco carcinogens in numerous cancers, there is a growing literature on the direct and indirect effects of smoking on treatment efficacy (short-term and long-term outcomes), toxicity and morbidity, quality of life (QOL), recurrence, second primary tumors (SPT), and survival time as summarized below. Oncology health professionals have called for increased advocacy for tobacco control. Despite the critical relevance of smoking to cancer outcomes, most oncology clinical trials do not collect data on smoking history and status unless the malignancy is widely acknowledged as smoking related (e.g., lung or head and neck cancer). Usually, these data are collected only at trial registration. Changes in smoking status during treatment or follow-up are monitored in very few trials and are infrequently reported in sample descriptions or included in analysis plans as a potential moderator of outcomes. Based on mounting evidence that tobacco use affects cancer treatment outcomes and survival, we recommend that smoking history and status be systematically collected as core data in all oncology clinical trials: at diagnosis, at trial registration, and throughout treatment and follow-up to long term survival or death. We feel that the inclusion and analysis of such data in clinical trials will add important information to the interpretation of outcomes and the development of scientific knowledge in this area. Smoking status has been called another "vital sign" because of its relevance to a patient's immediate medical condition. We explain the critical value of knowing the smoking status of every patient with cancer at every visit by providing a brief overview of the following research findings: (a) the effects of tobacco use on cancer treatment and outcome; (b) recent findings on the role of nicotine in malignant processes; (c) some unexpected results concerning tobacco status, treatment, and disease outcome; and (d) identifying key questions that remain to be addressed. We provide a suggested set of items for inclusion in clinical trial data sets that also are useful in clinical practice. PMID- 16214903 TI - MYB98 is required for pollen tube guidance and synergid cell differentiation in Arabidopsis. AB - The synergid cells of the female gametophyte play a role in many steps of the angiosperm fertilization process, including guidance of pollen tube growth to the female gametophyte. However, the mechanisms by which the synergid cells become specified and develop their unique features during female gametophyte development are not understood. We identified MYB98 in a screen for Arabidopsis thaliana genes expressed in the female gametophyte. MYB98 is a member of the R2R3-MYB gene family, the members of which likely encode transcription factors. In the context of the ovule, MYB98 is expressed exclusively in the synergid cells, and mutations in this gene affect the female gametophyte specifically. myb98 female gametophytes are affected in two unique features of the synergid cell, pollen tube guidance and the filiform apparatus, but are otherwise normal. MYB98 also is expressed in trichomes and endosperm. Homozygous myb98 mutants exhibit no sporophytic defects, including trichome and endosperm defects. Together, these data suggest that MYB98 controls the development of specific features within the synergid cell during female gametophyte development. PMID- 16214907 TI - Synthesizing the lifetime history of smoking. PMID- 16214908 TI - Modeling lung cancer risk in case-control studies using a new dose metric of smoking. AB - Many approaches have been taken to adjust for smoking in modeling cancer risk. In case-control studies, these metrics are often used arbitrarily rather than being based on the properties of the metric in the context of the study. Depending on the underlying study design, hypotheses, and base population, different metrics may be deemed most appropriate. We present our approach to evaluating different smoking metrics. We examine the properties of a new metric, "logcig-years", that we initially derived from using a biological model of DNA adduct formation. We compare this metric to three other smoking metrics, namely pack-years, square root pack-years, and a model in which smoking duration and intensity are separate variables. Our comparisons use generalized additive models and logistic regression to examine the relationship between the logit probability of cancer and each of the metrics, adjusting for other covariates. All models were fit using data from a lung cancer study of 1,275 cases and 1,269 controls that has focused on gene-smoking relationships. There was a very significant, linear relationship between logcig-years and the logit probability of lung cancer in this sample, without any need to adjust for smoking status. These properties together were not shared by the other metrics. In this sample, logcig-years captured more information about smoking that is important in lung cancer risk than the other metrics. In conclusion, we provide a general framework for evaluating different smoking metrics in studies where smoking is a critical variable. PMID- 16214909 TI - Vitamin D is associated with improved survival in early-stage non-small cell lung cancer patients. AB - Vitamin D may inhibit the development and progression of a wide spectrum of cancers. We investigated the associations of surgery season and vitamin D intake with recurrence-free survival (RFS) and overall survival in 456 early-stage non small cell lung cancer patients. The data were analyzed using log-rank test and Cox proportional hazards models. The median (range) follow-up time was 71 (0.1 140) months, with 161 recurrence and 231 deaths. Patients who had surgery in summer had a better RFS than those who had surgery in winter (adjusted hazard ratio, 0.75; 95% confidence interval, 0.56-1.01), with 5-year RFS rates of 53% (45-61%) and 40% (32-49%), respectively (P = 0.10, log-rank test). Similar association between surgery season and RFS was found among the 321 patients with dietary information (P = 0.33, log-rank test). There was no statistically significant association between vitamin D intake and RFS. Because both season and vitamin D intake are important predictors for vitamin D levels, we investigated the joint effects of surgery season and vitamin D intake. Patients who had surgery during summer with the highest vitamin D intake had better RFS (adjusted hazard ratio, 0.33; 95% confidence interval, 0.15-0.74) than patients who had surgery during winter with the lowest vitamin D intake, with the 5-year RFS rates of 56% (34-78%) and 23% (4-42%), respectively. Similar associations of surgery season and vitamin D intake with overall survival were also observed. In conclusion, the joint effects of surgery season and recent vitamin D intake seem to be associated with the survival of early-stage non-small cell lung cancer patients. PMID- 16214910 TI - Discrimination of breast cancer by anti-malignin antibody serum test in women undergoing biopsy. AB - PURPOSE: The anti-malignin antibody serum (AMAS) test (Oncolab, Boston, MA) has been reported as 97% sensitive and 95% specific for malignancies. To objectively assess accuracy of this test for discrimination of breast cancer, we studied a series of women undergoing core breast biopsy. SUBJECTS AND METHODS: Seventy-one core-needle breast biopsies were classified as malignant, suspicious, or benign by two independent pathologists blinded to AMAS results. Corresponding sera were read as AMAS positive, negative, or borderline by criteria used by Oncolab and also using criteria derived from receiver-operator curves based on values for slow (S-tag), fast (F-tag), and their difference (Net-tag) antibody reported by Oncolab. We calculated sensitivity and specificity and analyzed distributions by Fisher's exact test. RESULTS: Biopsies were read as 42 (59%) benign, 12 (17%) suspicious, and 17 (24%) malignant. By Oncolab criteria, sensitivity (59%) and specificity (62%) were maximized by pooling suspicious with malignant and AMAS borderline with positive (P = 0.098). Receiver-operator curves showed best sensitivity (62%) and specificity (69%) for the criterion AMAS positive if Net Tag > 135 microg/mL or S-Tag > 220 microg/mL (P = 0.015). CONCLUSIONS: The AMAS test discriminates suspicious and malignant from benign lesions, but sensitivity is insufficient to identify patients to be spared biopsy and false-positive rates are too high for population screening. PMID- 16214911 TI - Genetic variation in the growth hormone synthesis pathway in relation to circulating insulin-like growth factor-I, insulin-like growth factor binding protein-3, and breast cancer risk: results from the European prospective investigation into cancer and nutrition study. AB - Insulin-like growth factor-I (IGF-I) stimulates cell proliferation and can enhance the development of tumors in different organs. Epidemiologic studies have shown that an elevated level of circulating IGF-I is associated to increased risk of breast cancer as well as other cancers. Genetic variants affecting the release or biological action of growth hormone (GH), the main stimulator of IGF-I production, may predict circulating levels of IGF-I and have an effect on cancer risk. We tested this hypothesis with a large case-control study of 807 breast cancer patients and 1,588 matched control subjects nested within the European Prospective Investigation into Cancer and Nutrition. We genotyped 22 common single nucleotide polymorphisms in 10 genes involved in GH production and action (GHRH, GHRHR, SST, SSTR1-SSTR5, POU1F1, and GH1), and in parallel, we measured serum levels of IGF-I and IGFBP-3, its major binding protein, in samples of cases and controls. SST and SSTR2 polymorphisms showed weak but statistically significant associations with breast cancer risk. SSTR5 polymorphisms were associated with IGF-I levels, whereas one polymorphism in GHRHR and one in POU1F1 were associated with IGFBP-3 levels. Our conclusion is that common genetic variation in the GH synthesis pathway, as measured by single nucleotide polymorphisms selected in the present study, is not a major determinant of IGF-I and IGFBP-3 circulating levels, and it does not play a major role in altering breast cancer risk. PMID- 16214912 TI - Polymorphisms in DNA repair genes, medical exposure to ionizing radiation, and breast cancer risk. AB - An epidemiologic study was conducted to determine whether polymorphisms in DNA repair genes modify the association between breast cancer risk and exposure to ionizing radiation. Self-reported exposure to ionizing radiation from medical sources was evaluated as part of a population-based, case-control study of breast cancer in African-American (894 cases and 788 controls) and White (1,417 cases and 1,234 controls) women. Genotyping was conducted for polymorphisms in four genes involved in repair of radiation-induced DNA damage, the double-strand break repair pathway: X-ray cross-complementing group 3 (XRCC3) codon 241 Thr/Met, Nijmegen breakage syndrome 1 (NBS1) codon 185 Glu/Gln, X-ray cross-complementing group 2 (XRCC2) codon 188 Arg/His, and breast cancer susceptibility gene 2 (BRCH2) codon 372 Asn/His. Allele and genotype frequencies were not significantly different in cases compared with controls for all four genetic polymorphisms, and odds ratios for breast cancer were close to the null. Combining women with two, three, and four variant genotypes, a positive association was observed between breast cancer and number of lifetime mammograms (P(trend) < 0.0001). No association was observed among women with zero or one variant genotype (P = 0.86). Odds ratios for radiation treatments to the chest and number of lifetime chest X-rays were slightly elevated but not statistically significant among women with two to four variant genotypes. The study has several limitations, including inability to distinguish between diagnostic and screening mammograms or reliably classify prediagnostic mammograms and chest X-rays in cases. Prospective studies are needed to address whether common polymorphisms in DNA repair genes modify the effects of low-dose radiation exposure from medical sources. PMID- 16214913 TI - Associations between polymorphisms in the vitamin D receptor and breast cancer risk. AB - Biological and epidemiologic data suggest that vitamin D levels may influence breast cancer development. The vitamin D receptor (VDR) is a crucial mediator for the cellular effects of vitamin D and additionally interacts with other cell signaling pathways that influence cancer development. Because functional data exist on FOK1 and previous studies have suggested a relation between BSM1 and breast cancer risk, we evaluated the associations of the FOK1 and BSM1 VDR polymorphisms and breast cancer risk. In a case-control study nested within the Nurses' Health Study, we genotyped 1,234 incident cases (diagnosed between return of a blood sample in 1989-1990 and June 1, 2000) and 1,676 controls for FOK1, and 1,180 cases and 1,547 controls for BSM1. We observed a significantly increased risk of breast cancer among carriers of the ff genotype of FOK1 (multivariate odds ratio, 1.34; 95% confidence intervals, 1.06-1.69) compared with those with FF. We did not observe an association between polymorphisms in BSM1 and breast cancer risk (multivariate odds ratio, 0.93; 95% confidence intervals, 0.72-1.20) for BB versus bb). The FOK1 association did not vary significantly by menopausal status, estrogen, and progesterone receptor status of the tumors, or plasma levels of 25 hydroxyvitamin D or 1,25 dihydroxyvitamin D. Our results suggest that the VDR may be a mediator of breast cancer risk and could represent a target for cancer prevention efforts. PMID- 16214914 TI - Preventive health behaviors and familial breast cancer. AB - AIM: To examine medical and lifestyle preventive behaviors among women with varying levels of familial breast cancer risk. METHODS: Using cross-sectional data from the Minnesota Breast Cancer Family Study, a historical cohort of 426 families, we compared medical (mammography adherence, antiestrogen use, and prophylactic surgery) and lifestyle (physical activity, smoking, alcohol, and diet) behaviors across three groups of cancer-free women ages 18 to 95 defined by their family history of breast cancer. Family history was classified as high risk, moderate-risk, or average to low-risk depending on the number and degree of relationship of family members with breast cancer. RESULTS: After adjusting for age and education, high-risk women were twice as likely to have ever used an antiestrogenic agent (9.0% versus 4.6% among moderate-risk and 4.1% among average to low-risk; P = 0.002). Among women ages <40, the high-risk group were more likely to have ever had a mammogram (82% versus 47% among moderate-risk and 35% among average to low-risk; P < 0.001). Average to low-risk women were the least likely to be current smokers and high-risk women may consume slightly fewer fruits and vegetables compared with the other groups, but there were no other differences in lifestyle behaviors, including physical activity and alcohol use. CONCLUSIONS: Women with strong family histories of breast cancer are more likely to undertake medical but not lifestyle preventive behaviors. PMID- 16214915 TI - Effect of direct mail as a population-based strategy to increase mammography use among low-income underinsured women ages 40 to 64 years. AB - Women with inadequate health insurance have lower mammography rates than the general population. Finding successful strategies to enroll eligible women is an ongoing challenge for the National Breast and Cervical Cancer Early Detection Program. To test the effectiveness of a population-based strategy to increase mammography utilization among low-income underinsured women ages 40 to 64 years, a randomized trial was conducted to assess the effect of two mailed interventions on mammography utilization through Sage, the National Breast and Cervical Cancer Early Detection Program in Minnesota. Women (N = 145,467) ages 40 to 63 years [mean (SD), 49.7 (6.8)] with estimated household incomes below 50,000 US dollars (47.9% were < 35,000 US dollars) from a commercial database were randomized to three groups: Mail, Mail Plus Incentive, or Control. Both the Mail and the Mail Plus Incentive groups received two simple mailings prompting them to call a toll free number to access free mammography services. The Mail Plus Incentive intervention offered a small monetary incentive for a completed mammogram. After 1 year, both intervention groups had significantly higher Sage mammography rates than the Controls, and the Mail Plus Incentive group had a significantly higher rate than the Mail group. The Mail and Mail Plus Incentive interventions were estimated to produce increases in Sage screening rates of 0.23% and 0.75%, respectively, beyond the composite Control rate of 0.83%. Direct mail is an effective strategy for increasing mammography use through Sage. Coupling direct mail with an incentive significantly enhances the intervention's effectiveness. Direct mail should be considered as a strategy to increase mammography use among low-income, medically underserved women. PMID- 16214916 TI - Interaction of calcium supplementation and nonsteroidal anti-inflammatory drugs and the risk of colorectal adenomas. AB - BACKGROUND: Calcium and aspirin have both been found to be chemopreventive against colorectal neoplasia. However, the joint effect of the two agents has not been well investigated. METHODS: To explore the separate and joint effects of calcium and aspirin/nonsteroidal anti-inflammatory drugs (NSAID), we used data from two large randomized clinical trials among patients with a recent history of colorectal adenomas. In the Calcium Polyp Prevention Study, 930 eligible subjects were randomized to receive placebo or 1,200 mg of elemental calcium daily for 4 years. In the Aspirin/Folate Polyp Prevention Study, 1,121 eligible subjects were assigned to take placebo, 81 mg of aspirin, or 325 mg of aspirin daily for 3 years. In each study, subjects completed a validated food frequency questionnaire at enrollment and were asked periodically about medications and supplements used. Recurrent adenomas and advanced adenomas were the end points considered. We used generalized linear models to assess the separate and combined effects of aspirin (or NSAIDs) and calcium supplementation (or dietary calcium) and the interactions between these exposures. RESULTS: In the Calcium Trial, subjects randomized to calcium who also were frequent users of NSAIDs had a reduction of risk for advanced adenomas of 65% [adjusted risk ratio (RR), 0.35; 95% confidence interval (95% CI), 0.13-0.96], and there was a highly significant statistical interaction between calcium treatment and frequent NSAID use (P(interaction) = 0.01). Similarly, in the Aspirin Trial, 81 mg aspirin and calcium supplement use together conferred a risk reduction of 80% for advanced adenomas (adjusted RR, 0.20; 95% CI, 0.05-0.81); there was a borderline significant statistical interaction between the two treatments (P(interaction) = 0.09). In this trial, we found similar trends when we considered baseline dietary calcium intake instead of calcium supplements. For all adenomas considered together, the interactive patterns were not consistent. CONCLUSION: Data from two different randomized clinical trials suggest that calcium and NSAIDs may act synergistically to lower the risk of advanced colorectal neoplastic polyps. PMID- 16214917 TI - Does nonsteroidal anti-inflammatory drug use modify the effect of a low-fat, high fiber diet on recurrence of colorectal adenomas? AB - The Polyp Prevention Trial was designed to evaluate the effects of a high-fiber (18 g/1,000 kcal), high-fruit and -vegetable (3.5 servings/1,000 kcal), low-fat (20% energy) diet on recurrence of adenomatous polyps. Participants > or =35 years of age, with histologically confirmed colorectal adenoma(s) removed in the prior 6 months, were randomized to the intervention or control group. Demographic, dietary, and clinical information, including use of nonsteroidal anti-inflammatory drugs (NSAID), was collected at baseline and four annual visits. Adenoma recurrence was found in 754 of 1,905 participants and was not significantly different between groups. NSAID use was associated with a significant reduction in recurrence [odds ratio (OR), 0.77; 95% confidence interval (95% CI), 0.63-0.95]. In this analysis, NSAIDs modified the association between the intervention and recurrence at baseline (P = 0.02) and throughout the trial (P = 0.008). Among participants who did not use NSAIDs, the intervention was in the protective direction but did not achieve statistical significance (OR, 0.87; 95% CI, 0.69-1.09). The intervention was protective among males who did not use NSAIDs at baseline (OR, 0.71; 95% CI, 0.54-0.94), but not among NSAIDs users (OR, 1.09; 95% CI, 0.74-1.62). For females, corresponding OR estimates were 1.28 (95% CI, 0.86-1.90) and 2.30 (95% CI, 1.24-4.27), respectively. The protective association observed for NSAID use was stronger among control (OR, 0.63; 95% CI, 0.47-0.84) than for intervention group participants (OR, 0.97; 95% CI, 0.74 1.28). These results should be interpreted cautiously given that they may have arisen by chance in the course of examining multiple associations and Polyp Prevention Trial study participants were not randomly assigned to both dietary intervention and NSAID use. Nevertheless, our results suggest that adopting a low fat, high-fiber diet rich in fruits and vegetables may lower the risk of colorectal adenoma recurrence among individuals who do not regularly use NSAIDs. PMID- 16214918 TI - Genetic polymorphisms of flavin monooxygenase 3 in sulindac-induced regression of colorectal adenomas in familial adenomatous polyposis. AB - Sulindac is a nonsteroidal antiinflammatory drug with a chemopreventive effect in patients with familial adenomatous polyposis (FAP). In vivo, the active form of sulindac is sulindac sulfide, which is inactivated by the hepatic microsomal enzyme, flavin monooxygenase 3 (FMO3). In humans, numerous polymorphisms exist in FMO3, which alter enzymatic activity and subsequent substrate metabolism. We recently showed that certain polymorphic forms of FMO3 with reduced activity were associated with a more favorable response to sulindac in preventing the formation of adenomas in patients with FAP without polyps at baseline. Here, we determined whether these FMO3 polymorphisms correlated with the ability of sulindac to regress polyposis in patients with FAP who had polyps prior to treatment. Nineteen patients were treated with 150 mg sulindac twice a day for 6 months. The size and number of polyps in each patient was assessed at baseline (prior to the administration of sulindac), and at 3 and 6 months. Genotyping was done on seven established FMO3 polymorphisms with functional significance-M66I, E158K, P153L, V257M, E305X, E308G, and R492W. Statistical analyses were done with Wilcoxon rank sum test. Of the loci examined, only E158K and E308G showed polymorphic changes. Six patients exhibited polymorphisms in both E158K and E308G loci and were designated as genotype combination 1. The remaining patients were designated as genotype combination 2. Over the course of treatment, patients with genotype combination 1 had a greater reduction in both the size and number of polyps than those with genotype combination 2. These results suggest that combined polymorphic changes in the E158K and E308G alleles may protect against polyposis in patients with FAP treated with sulindac. PMID- 16214919 TI - Expression of vitamin D receptor and 25-hydroxyvitamin D3-1{alpha}-hydroxylase in normal and malignant human colon. AB - Considerable evidence exists to support the use of vitamin D to prevent and/or treat colorectal cancer. However, the routine use of bioactive vitamin D, 1,25 dihydroxyvitamin D3, is limited by the side effect of toxic hypercalcemia. Recent studies, however, suggest that colonic epithelial cells express 25-hydroxyvitamin D3-1alpha-hydroxylase, an enzyme that converts nontoxic pro-vitamin D, 25 hydroxycholecalciferol [25(OH)D3], to its bioactive form. Yet, nothing is known as to the cellular expression of 1alpha-hydroxylase and the vitamin D receptor (VDR) in the earliest histopathologic structures associated with malignant transformation such as aberrant crypt foci (ACF) and polyps [addressing the possibility of using nontoxic 25(OH)D3 for chemoprevention], nor is anything known as to the expression of these proteins in colorectal cancer as a function of tumor cell differentiation or metastasis [relevant to using 25(OH)D3 for chemotherapy]. In this study, we show that 1alpha-hydroxylase is present at equal high levels in normal colonic epithelium as in ACFs, polyps, and colorectal cancer irrespective of tumor cell differentiation. In contrast, VDR levels were low in normal colonic epithelial cells; were increased in ACFs, polyps, and well differentiated tumor cells; and then declined as a function of tumor cell de differentiation. Both 1alpha-hydroxylase and VDR levels were negligible in tumor cells metastasizing to regional lymph nodes. Overall, these data support using 25(OH)D3 for colorectal cancer chemoprevention but suggest that pro-vitamin D is less likely to be useful for colorectal cancer chemotherapy. PMID- 16214921 TI - Melanocortin-1 receptor (MC1R) gene variants and dysplastic nevi modify penetrance of CDKN2A mutations in French melanoma-prone pedigrees. AB - Germline mutations in CDKN2A gene predispose to melanoma with high but incomplete penetrance. Penetrance of CDKN2A gene was found to be significantly influenced by host factors (nevus phenotypes and sunburn) on one hand and by variants of MC1R gene (RHC variants consistently associated with red hair and fair skin) on the other hand. Our goal was to examine the joint effects of MC1R variants and other potential risk factors [total nevi, dysplastic nevi, pigmentary traits (skin, hair and eye color), skin reactions to sunlight, and degree of sun exposure] on CDKN2A penetrance. Clinical, genetic, and covariate data were recorded in 20 French melanoma-prone families with cosegregating CDKN2A mutations. Analysis of the cotransmission of melanoma and CDKN2A mutations was conducted by likelihood based methods using the regressive logistic models, which can account for a variation of disease risk with age and can include the aforementioned risk factors as covariates. RHC variants, considered either alone or in the presence of pigmentation and nevus phenotypes, were found to increase significantly CDKN2A penetrance. Multivariate analysis, using a stepwise selection procedure, showed significant effects of two factors on melanoma risk in CDKN2A mutations carriers: RHC variants [odds ratio of hazard function (OR), 2.21; P = 0.03] and dysplastic nevi (OR, 2.93; P < 0.01). Such results may have important consequences to improve the prediction of melanoma risk in families. PMID- 16214920 TI - Low-fat, high fruit and vegetable diets and weight loss do not affect biomarkers of cellular proliferation in Barrett esophagus. AB - Risk factors for esophageal adenocarcinoma include obesity, high fat intake, and low consumption of fruits and vegetables. This trial tested whether an intervention to reduce these risk factors in patients with Barrett esophagus, a preneoplastic condition for esophageal adenocarcinoma, could reduce biomarkers of cellular proliferation and, by inference, the risk of neoplastic progression. Eighty-seven men and women with Barrett esophagus were randomized to an intensive dietary intervention or control group. At baseline, 18 and 36 months after intervention, biopsies were obtained at 2-cm intervals throughout the length of the Barrett segment. Ki67/DNA content flow cytometry was used to assess (a) % Ki67-positive proliferating diploid G(1) cells, (b) % total Ki67-positive proliferating cells, (c) presence of aneuploidy, and (d) presence of >6% of cells in the 4N (G(2)/tetraploid) fraction of the cell cycle. We also assessed re epithelialization and length of the Barrett segment, reflux symptoms, and medication use. The intervention effects for energy, fat, fruits and vegetables, and weight were, respectively, -314 kcal, -12.2% energy, 1.8 servings/d, and -4.0 kg at 18 months (all P < 0.005) and were smaller but remained significant at 36 months. There were no significant effects of the intervention on any biomarker of cellular proliferation. The intervention effects +/- SE for mean %G(1) Ki67+ cells were 0.98 +/- 1.58 at 18 months and 1.79 +/- 1.31 at 36 months; the relative risks (95% confidence interval) for developing >6% of cells in 4N were 0.5 (0.1-2.6) at 18 months and 0.75 (0.2-3.1) at 36 months. A single control participant developed aneuploidy. There were no significant effects on re epithelialization, segment length, or reflux medication use. We conclude that substantial dietary change has no short-term effects on biomarkers of cellular proliferation in Barrett esophagus or on clinical observations of the Barrrett segment. PMID- 16214922 TI - Polymorphisms and haplotypes in the cytochrome P450 17A1, prolactin, and catechol O-methyltransferase genes and non-Hodgkin lymphoma risk. AB - Expression of prolactin and of prolactin and estrogen receptors in lymphocytes, bone marrow, and lymphoma cell lines suggests that hormonal modulation may influence lymphoma risk. Prolactin and estrogen promote the proliferation and survival of B cells, factors that may increase non-Hodgkin lymphoma risk, and effects of estrogen may be modified by catechol-O-methyltransferase (COMT), an enzyme that alters estrogenic activity. Cytochrome P450 17A1 (CYP17A1), a key enzyme in estrogen biosynthesis, has been associated with increased cancer risk and may affect lymphoma susceptibility. We studied the polymorphisms prolactin (PRL) -1149G>T, CYP17A1 -34T>C, and COMT 108/158Val>Met, and predicted haplotypes among a subset of participants (n = 308 cases, n = 684 controls) in a San Francisco Bay Area population-based non-Hodgkin lymphoma study (n = 1,593 cases, n = 2,515 controls) conducted from 1988 to 1995. Oral contraceptive and other hormone use also was analyzed. Odds ratios (OR) for non-Hodgkin lymphoma and follicular lymphoma were reduced for carriers of the PRL -1149TT genotype [OR, 0.64; 95% confidence interval (95% CI), 0.41-1.0; OR, 0.53; 95% CI, 0.26-1.0, respectively]. Diffuse large-cell lymphoma risk was increased for those with CYP17A1 polymorphisms including CYP17A1 -34CC (OR, 2.0; 95% CI, 1.1-3.5). ORs for all non-Hodgkin lymphoma and follicular lymphoma among women were decreased for COMT IVS1 701A>G [rs737865; variant allele: OR, 0.53; 95% CI, 0.34-0.82; OR, 0.42; 95% CI, 0.23-0.78, respectively]. Compared with never users of oral contraceptives, a 35% reduced risk was observed among oral contraceptive users in the total population. Reduced ORs for all non-Hodgkin lymphoma were observed with use of exogenous estrogens among genotyped women although 95% CIs included unity. These results suggest that PRL, CYP17A1, and COMT may be relevant genetic loci for non-Hodgkin lymphoma and indicate a possible role for prolactin and estrogen in lymphoma pathogenesis. PMID- 16214923 TI - Familial aggregation and heterogeneity of non-Hodgkin lymphoma in population based samples. AB - The importance of genetic factors in the etiology of non-Hodgkin lymphoma (NHL) is suggested by case-control and cohort studies. Most previous studies have been too small to estimate accurately risks of specific categories of lymphoproliferative malignancies in relatives of NHL cases or to quantify the contribution of NHL case characteristics to familial risk. We have overcome sample size limitations and potential recall bias by using large databases from Sweden and Denmark. Diagnoses of lymphoproliferative malignancies were compared in 70,006 first-degree relatives of 26,089 NHL cases (including 7,432 with subtype information) versus 161,352 first-degree relatives of 58,960 matched controls. Relatives of NHL cases were at significantly increased risk for NHL [relative risk (RR), 1.73; 95% confidence interval (95% CI), 1.39-2.15], Hodgkin lymphoma (RR, 1.41; 95% CI, 1.0-1.97), and nonsignificantly for chronic lymphocytic leukemia (CLL; RR, 1.31; 95% CI, 0.93-1.85). No increased risk was found for multiple myeloma among case relatives. Findings with respect to siblings compared with parents and offspring or with respect to age at diagnosis of proband were inconsistent. In both populations, relatives of cases with an aggressive NHL subtype were at substantially increased risk of NHL (combined RR, 3.56; 95% CI, 1.80-7.02). We conclude that NHL has an important familial component, which is shared with Hodgkin lymphoma and CLL. We estimate that the absolute lifetime risk for a first-degree relative of an NHL case to develop NHL is 3.6% (compared with a population risk of 2.1%) and higher if the index case had an aggressive subtype of NHL. PMID- 16214924 TI - Association of polymorphisms in ERCC2 gene with non-familial thyroid cancer risk. AB - The ERCC2 protein is an evolutionary conserved ATP-dependent helicase that is associated with a TFIIH transcription factor complex and plays an important role in nucleotide excision repair. Mutations in this gene are responsible for xeroderma pigmentosum and also for Cocayne syndrome and trichothiodystrophy. Several single nucleotide polymorphisms have been identified in the ERCC2 locus. Among them, a G23591A polymorphism in the codon 312 results in an Asp --> Asn substitution in a conserved region and a A35931C polymorphism in the codon 751 results in a Lys --> Gln substitution. Because these polymorphisms have been associated with an increased risk for several types of cancers, we carried out an hospital based case-control study in a Caucasian Portuguese population to evaluate the potential role of these polymorphisms on the individual susceptibility to thyroid cancer. The results obtained did not reveal a significant association between each individual polymorphism studied (G23591A and A35931C) and an increased thyroid cancer risk, but individuals homozygous for non wild-type variants are overrepresented in patients group. The evaluation of the different haplotypes generated by these polymorphisms showed that individuals simultaneously homozygous for rare variants of both polymorphisms have an increased risk for thyroid cancer [adjusted odds ratio (OR) 3.084; 95% confidence interval (95% CI), 1.347-7.061; P = 0.008] and for papillary thyroid-type tumors (adjusted OR, 2.997; 95% CI, 1.235-7.272; P = 0.015) but not for follicular thyroid-type tumors. These results suggest that genetic polymorphisms in this gene might be associated with individual susceptibility towards thyroid cancer, mainly papillary-type tumors, but larger studies are required to confirm these results. PMID- 16214925 TI - Circulating levels of inflammatory markers and cancer risk in the health aging and body composition cohort. AB - BACKGROUND: Chronic inflammation is associated with processes that contribute to the onset or progression of cancer. This study examined the relationships between circulating levels of the inflammatory markers interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor-alpha (TNF-alpha) and total as well as site-specific cancer incidence. METHODS: Study subjects (n = 2,438) were older adults (ages 70-79 years) participating in the Health Aging and Body Composition study, who did not report a previous cancer diagnosis (except for nonmelanoma skin cancer) at baseline. Incident cancer events (n = 296) were ascertained during an average follow-up of 5.5 years. Inflammatory markers were measured in stored baseline fasting blood samples. RESULTS: The adjusted hazard ratios (95% confidence intervals) for incident cancer associated with a 1-unit increase on the natural log-scale were 1.13 (0.94-1.37), 1.25 (1.09-1.43), and 1.28 (0.96 1.70) for IL-6, CRP, and TNF-alpha, respectively. Markers were more strongly associated with cancer death: hazard ratios were 1.63 (1.19-2.23) for IL-6, 1.64 (1.20-2.24) for CRP, and 1.82 (1.14-2.92) for TNF-alpha. Although precision was low for site-specific analyses, our results suggest that all three markers were associated with lung cancer, that IL-6 and CRP were associated with colorectal cancer, and that CRP was associated with breast cancer. Prostate cancer was not associated with any of these markers. CONCLUSIONS: These findings suggest that (a) the associations between IL-6, CRP, and TNF-alpha and the risk of cancer may be site specific and (b) increased levels of inflammatory markers are more strongly associated with the risk of cancer death than cancer incidence. PMID- 16214926 TI - Toenail arsenic concentrations, GSTT1 gene polymorphisms, and arsenic exposure from drinking water. AB - Toenail arsenic (As) concentrations were evaluated as a biomarker of inorganic As (As(in)) exposure in a population residing in an As-endemic region of Bangladesh. Drinking water and toenail samples were collected from 48 families (n = 223) every 3 months for 2 years and analyzed for As using inductively coupled plasma mass spectrometry. Drinking water collected 3, 6, and 9 months before each toenail sample collection was combined into a weighted lagged exposure variable. The contribution of each water sample to the measured toenail As concentration was estimated using maximum likelihood that accounted for fluctuations in drinking water exposure and toenail growth. The best model attributed 69%, 14%, and 17% of the toenail As content to drinking water exposures that occurred 3, 6, and 9 months before toenail collection [95% confidence intervals (95% CI), 0.46 0.97, 0.00-0.31, and 0.03-0.35, respectively]. Generalized additive mixed models using penalized regression splines were employed to model the data. Below a drinking water concentration of 2 mug As/L, no relationship between drinking water As and toenail As concentrations was observed. Above this concentration, toenail As content increased in a dose-dependent fashion as drinking water As increased. Age was a significant effect modifier of drinking water As exposure on toenail As (beta = 0.01; 95% CI, 0.002-0.02). Individuals possessing GSTT1-null genotypes had significantly more As in their toenails in contrast to GSTT1 wild type individuals (beta = 0.11; 95% CI, 0.06-0.2). Therefore, it seems that GSTT1 modifies the relationship between As(in) exposure and toenail As(in) content. PMID- 16214927 TI - Reliability and validity of a telephone questionnaire for estimating lifetime personal sun exposure in epidemiologic studies. AB - Our Australia-wide case-control study of ocular melanoma diagnosed in 1996 to 1997 needed a short telephone interview on sun exposure. We constructed one by examining data from 700 controls ages 40 to 64 years in the Geraldton Skin Cancer Survey in 1988; they had answered a "whole-of-life" questionnaire in a face-to face interview. Sun exposure in their first 4 decade years of age best predicted their lifetime annual average sun exposure, so the shortened questionnaire asked about sun exposure in these 4 decade years only. Retesting 60 participants 1 year later with the whole-of-life questionnaire gave an intraclass correlation coefficient of 0.65 (95% confidence interval, 0.48-0.78) for ranked total sun exposure between the two interviews; the intraclass correlation coefficient was higher in men (0.73) than in women (0.54). Correlations were also high between parallel measurements of sun exposure on working days in the decade years and in outdoor occupations throughout life in the telephone interview of the ocular melanoma study (Spearman's R = 0.75) and in another study of non-Hodgkin's lymphoma (R = 0.71). Agreement between simultaneous parallel measures of total exposure (nonworking + working day and recreational + occupational exposure) was slightly weaker and of nonworking day and recreational exposure much weaker. Occupational exposure in women was much less strongly correlated with total exposure than it was in men possibly because of their frequently combined work and family roles, which the questionnaires did not try to separate. Research is needed into how this might be done to improve sun exposure measurement in women. PMID- 16214928 TI - Body mass index and serum 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane in nulliparous Chinese women. AB - BACKGROUND: Basic health indicators, such as body mass index (BMI), have been associated with serum 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane/1,1-dichloro 2,2-bis(p-chlorophenyl)ethylene (DDT/DDE) levels; however, both positive and inverse associations of BMI with serum DDT/DDE have been reported. Given the association of BMI with a number of outcomes, it may confound studies of DDT/DDE associated health effects. We investigated the relationship of BMI with serum DDT/DDE accounting for other determinants of exposure among women with relatively recent environmental exposures to DDT. METHODS: Serum DDT/DDE was analyzed in 466 nonsmoking, nulliparous women recruited from Anhui province in China between 1996 and 1998 as part of a reproductive health study of textile workers. The women in the sample were born between 1963 and 1977, 8 to 21 years before China's 1984 DDT ban. We used multivariate linear regression to investigate associations of BMI, age, and birth year with serum DDT/DDE. RESULTS: Mean (SD) serum total DDT concentration was 32 ng/g (17.8 ng/g). Birth year showed an inverse relationship with serum DDT independent of age. Despite limited variability in BMI, there was a consistent inverse relationship between BMI and serum DDT. Specifically, each kg/m(2) increase in BMI was associated with a -1.34 ng/g (95% confidence interval, -2.12 to -0.56 ng/g) decrease in serum total DDT. CONCLUSIONS: There were high total DDT levels in this sample of nulliparous Chinese women relative to Western populations, birth year was more strongly associated with serum DDT than age, and BMI was inversely related to serum DDT in this study. PMID- 16214929 TI - Prevalence and clustering patterns of human papillomavirus genotypes in multiple infections. AB - Prevalence of multiple human papillomavirus (HPV) infections, involvement of specific HPV phylogenetic clades in multiple infections, and clustering patterns of multiple infections at the clade level were assessed in 854 HIV (-) and 275 HIV (+) women cross-sectionally. Reverse line blot assay was used to detect 27 HPV genotypes. Involvement of specific clades in coinfections and clustering patterns were assessed using HPV clade/genotype as the unit of analyses. Expected frequencies assuming independence for all possible clade combinations in two genotype infections were derived using a multinomial expansion and comparisons of observed and expected frequencies were done using a composite goodness-of-fit test. In all, 100 two-genotype infections were detected; 61 in HIV (-) and 39 in HIV (+) women. Clade A9 (HPV types 16, 31, 33, 35, 52, and 58) was significantly less likely to be involved in multiple infections compared with all other clades (55.2% versus 64.6%; adjusted odds ratios, 0.68; 95% confidence interval, 0.48 0.95). Observed patterns for all possible clade combinations (among HPV clades A3, A5, A6, A7, A9, and A10) in two-genotype infections did not significantly differ from those expected in the entire sample, across HIV, Pap smear, and age strata (all goodness-of-fit exact P > 0.20). These results indicate that clade A9 is less likely to be involved in multiple infections and that HPV genotypes predominantly establish multiple infections at random, with little positive/negative clustering for either phylogenetically related or unrelated types. The current method of analysis affords the opportunity to test clustering of a large number of HPV genotype/clade combinations at nominal alpha levels. PMID- 16214930 TI - Positive association of farm or rural residence with acute myeloid leukemia incidence in a cohort of older women. AB - The etiology of acute myeloid leukemia (AML) is relatively unknown. Incidence rates are highest in the agricultural Midwest region compared with other regions of the United States. Many studies have examined the relationship between farming and leukemia, but most have mainly focused on men. We examined the potential association between farm or rural residence and AML in the Iowa Women's Health Study. In 1986, 37,693 women who were free of prior cancer completed a lifestyle and health questionnaire, which included a question on the place of residence. Women were subsequently followed until 2002 for cancer incidence; 79 women developed AML during the time period. Women who lived on a farm at baseline were more likely (relative risk, 1.91; 95% confidence interval, 1.19-3.05) to develop AML compared with women who did not live on a farm. Further, women who reported living on a farm or in a rural area were twice as likely (relative risk, 2.38; 95% confidence interval, 1.33-4.26) to develop AML compared with women who lived in a city with a population of >10,000 people. These results provide evidence that women who live on farms or rural areas are at an increased risk of AML. PMID- 16214931 TI - Genetic variation and willingness to participate in epidemiologic research: data from three studies. AB - The differences in common genetic polymorphism frequencies by willingness to participate in epidemiologic studies are unexplored, but the same threats to internal validity operate as for studies with nongenetic information. We analyzed single nucleotide polymorphism genotypes, haplotypes, and short tandem repeats among control groups from three studies with different recruitment designs that included early, late, and never questionnaire responders, one or more participation incentives, and blood or buccal DNA collection. Among 2,955 individuals, we compared 108 genotypes, 8 haplotypes, and 9 to 15 short tandem repeats by respondent type. Among our main comparisons, single nucleotide polymorphism genotype frequencies differed significantly (P < 0.05) between respondent groups in six instances, with 13 expected by chance alone. When comparing the odds of carrying a variant among the various response groups, 19 odds ratios were /=1.40, levels that might be notably different. Among the various respondent group comparisons, haplotype and short tandem repeat frequencies were not significantly different by willingness to participate. We observed little evidence to suggest that genotype differences underlie response characteristics in molecular epidemiologic studies, but a greater variety of genes should be examined, including those related to behavioral traits potentially associated with willingness to participate. To the extent possible, investigators should evaluate their own genetic data for bias in response categories. PMID- 16214932 TI - No association between EGF gene polymorphism and gastric cancer. AB - The etiology of gastric cancer is not well-understood. Epidermal growth factor (EGF) transduces growth signals to mitogen-activated protein kinase via RAS and BRAF, and EGF/EGF receptor interaction is important for tumor growth and progression. Previous studies have reported that the EGF +61 (A/G) single nucleotide polymorphism in the 5'-untranslated region of the EGF gene is functional, and is associated with gastric cancer and various malignancy. Individuals with the EGF A/A genotype produce less EGF than individuals with G/G or G/A. We investigated a single nucleotide polymorphism at exon 1 of EGF, named rs4444903 in NCI dbSNP, in 454 Japanese subjects undergoing a health checkup and 202 patients with gastric cancer. Genotype was determined by PCR with confronting two-pair primers. Results showed that EGF polymorphism was not associated with gastric cancer but that the EGF A/A genotype showed a protective effect (odds ratios, 0.58; 95% confidence interval, 0.29-1.17 relative to G/G). Furthermore, when we divided cases into two groups, a differentiated type and an undifferentiated type, the A/A and G/A combined was found to be lower frequency in the latter type than in the former type without significance (OR, 0.81; 95% confidence interval, 0.44-1.49 relative to G/G). As is the case with any malignancy, other factors are involved, including environmental and host factors. The present results show that although EGF is necessary for cancer, it is not sufficient. We also found ethnic heterogeneity in the functional EGF polymorphism. Because the relationship between EGF polymorphism and malignancy remains inconsistent, confirmation of the role of EGF polymorphism in gastric cancer requires a much larger study. PMID- 16214933 TI - Lack of association between -251 T>A polymorphism of IL8 and lung cancer risk. PMID- 16214934 TI - No association between glutathione peroxidase Pro198Leu polymorphism and breast cancer risk. PMID- 16214935 TI - No association between genetic polymorphisms in insulin and insulin receptor substrate-1 and prostate cancer. PMID- 16214936 TI - Thyroid hormone in health and disease. AB - Thyroid disease is common, affecting around 2% of women and 0.2% of men in the UK. Our understanding of the effects of thyroid hormones under physiological circumstances, as well as in pathological conditions, has increased dramatically during the last two centuries and it has become clear that overt thyroid dysfunction is associated with significant morbidity and mortality. Both hypo-and hyperthyroidism and their treatments have been linked with increased risk from cardiovascular disease and the adverse effects of thyrotoxicosis in terms of osteoporosis risk are well established. Although the evidence suggests that successful treatment of overt thyroid dysfunction significantly improves overall survival, the issue of treating mild or subclinical hyper- and hypothyroidism remains controversial. Furthermore, the now well-established effects of thyroid hormones on neurodevelopment have sparked a whole new debate regarding the need to screen pregnant women for thyroid function abnormalities. This review describes the current evidence of the effects of thyroid hormone on the cardiovascular, skeletal and neurological systems, as well as the influence of thyroid diseases and their treatments on the development of malignancy. Furthermore we will describe some recent developments in our understanding of the relationship between thyroid status and health. PMID- 16214937 TI - Autoinduction of tumor necrosis factor-alpha in FRTL-5 rat thyroid cells. AB - Tumor necrosis factor-alpha (TNFalpha) may play a role in the development of autoimmune thyroiditis such as Hashimoto's thyroiditis. In the present study, we examined whether TNFalpha induced its own expression in FRTL-5 rat thyroid cells. Lipopolysaccharide (LPS) markedly increased TNFalpha mRNA levels in FRTL-5 cells as assessed by semiquantitative RT-PCR. In addition, LPS-stimulated cells released TNFalpha protein into the culture medium. Similarly, TNFalpha induced its own gene and protein expression in FRTL-5 cells as assessed by RT-PCR and metabolic labeling and immunoprecipitation of TNFalpha. The autoinduction of TNFalpha gene was also observed in TNFalpha-stimulated human thyroid epithelial cells. TNFalpha induction was specific to LPS and TNFalpha since interferon-alpha or amiodarone failed to increase TNFalpha mRNA levels in FRTL-5 cells. Human TNFalpha induced rat TNFalpha gene expression, indicating that type 1 TNF receptor (TNF-R) is involved in the autoinduction. TNFalpha did not increase either type 1 or type 2 TNF-R mRNA levels, suggesting that upregulation of TNF receptors is not involved in the autoinduction of TNFalpha. Although the biological significance of autoinduction of TNFalpha remains unclear, our results suggest that thyroid epithelial cells may participate in the development of autoimmune thyroiditis through production of TNFalpha. Furthermore, inhibition of TNFalpha production in the thyroid may represent a novel approach to mitigating inflammation in autoimmune thyroiditis. PMID- 16214938 TI - STAT5 activation by human GH protects insulin-producing cells against interleukin 1beta, interferon-gamma and tumour necrosis factor-alpha-induced apoptosis independent of nitric oxide production. AB - The proinflammatory cytokines interleukin-1beta (IL-1beta), interferon-gamma (IFN gamma) and tumour necrosis factor-alpha (TNF-alpha) are toxic to pancreatic beta cells and are implicated in the pathogenesis of type 1 diabetes. We have previously found that GH and prolactin (PRL) stimulate both proliferation and insulin production in pancreatic beta-cells and rat insulin-producing INS-1 cells. Here we report that human (h) GH can prevent the apoptotic effects of IL 1beta, IFN-gamma and TNF-alpha in INS-1 and INS-1E cells. Using adenovirus mediated gene transfer, we found that the anti-apoptotic effect of hGH is abrogated by expression of a dominant negative signal transducer and activator of transcription (STAT5) mutant in INS-1E cells. hGH and the cytotoxic cytokines was found to additively increase suppressor of cytokine signalling-3 mRNA expression after 4 h of exposure. In order to identify possible targets for the STAT5 mediated protection of INS-1E cells, we studied the effect of hGH on activation of the transcription factors STAT1 and nuclear factor-kappaB (NF-kappaB) by IFN gamma and IL-1beta+TNF-alpha respectively. Gel retardation experiments showed that hGH affects neither IFN-gamma+TNF-alpha-induced STAT1 DNA binding nor IL 1beta and IFN-gamma+TNF-alpha-induced NFkappaB DNA binding. The lack of influence of hGH on cytokine-mediated activation of STAT1 and NFkappaB is in accordance with the finding that hGH had only a minor effect on cytokine-induced inducible nitric oxide synthase (iNOS) gene expression and in fact augmented the IL-1beta stimulated nitric oxide production. As the anti-apoptotic Bcl-xL gene has been shown to harbour a STAT5-binding element we measured the expression of Bcl-xL as well as the pro-apoptotic Bax. We found that hGH increased the Bcl-xL/Bax ratio both in the absence and in the presence of cytotoxic cytokines. In conclusion, these results suggested that GH and PRL protect beta-cells against cytotoxic cytokines via STAT5-dependent mechanisms distal to iNOS activation possibly at the level of Bcl-xL. PMID- 16214939 TI - Oxidative and nitrosative stress in brain mitochondria of diabetic rats. AB - Diabetic encephalopathy, characterized by impaired cognitive functions and neurochemical and structural abnormalities, may involve direct neuronal damage caused by intracellular glucose. The study assesses the direct effect of chronic hyperglycemia on the function of brain mitochondria, the major site of reactive species production, in diabetic streptozotocin (STZ) rats. Oxidative stress plays a central role in diabetic tissue damage. Alongside enhanced reactive oxygen species (ROS) levels, both nitric oxide (NO) levels and mitochondrial nitric oxide synthase expression were found to be increased in mitochondria, whereas glutathione (GSH) peroxidase activity and manganese superoxide dismutase protein content were reduced. GSH was reduced and GSH disulfide (GSSG) was increased in STZ rats. Oxidative and nitrosative stress, by reducing the activity of complexes III, IV and V of the respiratory chain and decreasing ATP levels, might contribute to mitochondrial dysfunction. In summary, this study offers fresh evidence that, besides the vascular-dependent mechanisms of brain dysfunction, oxidative and nitrosative stress, by damaging brain mitochondria, may cause direct injury of neuronal cells. PMID- 16214940 TI - Differential effect of inbred mouse strain (C57BL/6, DBA/2, 129T2) on insulin secretory function in response to a high fat diet. AB - The increasing production of genetically-modified mouse models has necessitated studies to determine the inherent physiological characteristics of commonly used mouse strains. In this study we examined insulin secretory function in response to an intravenous bolus of glucose or glucose plus arginine in anesthetized C57BL/6, DBA/2 and 129T2 mice fed either a control or high fat diet for 6 weeks. The results show that 129T2 mice had higher fasting plasma glucose levels and lower fasting plasma insulin levels compared with C57BL/6 and DBA/2 mice regardless of diet. Furthermore, 129T2 mice were glucose intolerant and secreted significantly less insulin in response to glucose and glucose plus arginine irrespective of diet compared with the other two strains of mice. DBA/2 mice hypersecreted insulin in response to glucose and glucose plus arginine compared with C57BL/6 and 129T2 mice. Moreover while first phase insulin secretion was appropriately increased in response to the high fat diet in C57BL/6 and 129T2 mice, this was not the case for DBA/2 mice. Mean islet area was decreased in response to a high fat diet in DBA/2 mice, while there was no dietary effect on the other two strains. This study highlights the inherent genetic differences that exist among seemingly normal strains of mice that are commonly used to make transgenic and knockout mice. Understanding these differences will provide researchers with the information to choose the appropriate genetic background on which to express their particular genetic alteration. PMID- 16214941 TI - cDNA structure of an insulin-related peptide in the Pacific oyster and seasonal changes in the gene expression. AB - Insulin-related peptide cDNA was characterized in the Pacific oyster Crassostrea gigas. It was determined that three transcripts with differing lengths of 3' untranslated region (3'-UTR) were expressed in the visceral ganglia. The insulin related peptide cDNA contained a number of AUUUA motifs that were typical of adenylate/uridylate-rich elements in the 3'-UTR. The deduced preprohormone was a polypeptide of 161 residues and showed a conformation typical of preprohormones of the insulin superfamily, which included conserved amino acids necessary to adopt the globular insulin structure. The expression of the three different transcripts was variable throughout the year, with the highest expression observed in March and lower expression in November and July. The expression of the shortest mRNA in March was about tenfold higher than in July, while the expression of the longest transcript varied approximately twofold during the year. The accumulation of glycogen in the soft body rapidly increased in October and November, and robust body growth and gametogenetic development occurred in March to May. The period of the highest expression of the oyster insulin-related peptide gene corresponded to the onset of body growth and gametogenetic development, but did not overlap with the period of glycogen accumulation. This is the first report that fully details the structure and expression of the insulin-related peptide gene in bivalves. PMID- 16214942 TI - Nutritional status affects 20-hydroxyecdysone concentration and progression of oogenesis in Drosophila melanogaster. AB - Drosophila egg production depends upon the nutritional available to females. When food is in short supply, oogenesis is arrested and apoptosis of the nurse cells is induced at mid-oogenesis via a mechanism that is probably controlled by ecdysteroid hormone. We have shown that expression of some ecdysone-response genes is correlated with apoptosis of egg chambers. Moreover, ecdysteroid injection and application of juvenile hormone induces and suppresses the apoptosis, respectively. In this study, we investigated which tissues show increases in the concentration of ecdysteroids under nutritional shortage to begin to link together nutrient intake, hormone regulation and the choice between egg development or apoptosis made within egg chambers. We measured ecdysteroid levels in the whole body, ovaries and haemolymph samples by RIA and found that the concentration of ecdysteroid increased in all samples. This contributes to the idea that nutritional shortage leads to a rapid high ecdysteroid concentration within the fly and that the high concentration induces apoptosis. Low concentrations of ecdysteroid are essential for normal oogenesis. We suggest there is threshold concentration in the egg chambers and that apoptosis at mid oogenesis is induced when the ecdysteroid levels exceed the threshold. Starvation causes the ovary to retain the ecdysteroid it produces, thus enabling individual egg chambers to undergo apoptosis and thus control the number of eggs produced in relation to food intake. PMID- 16214943 TI - Tissue-specific effects of leptin administration on the abundance of mitochondrial proteins during neonatal development. AB - Many tissues undergo a rapid transition after birth, accompanied by dramatic changes in mitochondrial protein function. In particular, uncoupling protein (UCP) abundance increases at birth in the lung and adipose tissue, to then gradually decline, an adaptation that is important in enabling normal tissue function. Leptin potentially mediates some of these changes and is known to promote the loss of UCP1 from brown fat but its effects on UCP2 and related mitochondrial proteins (i.e. voltage-dependent anion channel (VDAC) and cytochrome c) in other tissues are unknown. We therefore determined the effects of once-daily jugular venous administration of ovine recombinant leptin on mitochondrial protein abundance as determined by immunoblotting in tissues that do (i.e. the brain and pancreas) and do not (i.e. liver and skeletal muscle) express UCP2. Eight pairs of 1-day-old lambs received either 100 mug leptin or vehicle daily for 6 days, before tissue sampling on day 7. Administration of leptin diminished UCP2 abundance in the pancreas, but not the brain. Leptin administration had no affect on the abundance of VDAC or cytochrome c in any tissue examined. In leptin-administered animals, but not controls, UCP2 abundance in the pancreas was positively correlated with VDAC and cytochrome c content, and UCP2 abundance in the brain with colonic temperature. In conclusion, leptin administration to neonatal lambs causes a tissue-specific loss of UCP2 from the pancreas. These effects may be important in the regulation of neonatal tissue development and potentially for optimising metabolic control mechanisms in later life. PMID- 16214944 TI - Regulated expression of putative membrane progestin receptor homologues in human endometrium and gestational tissues. AB - Rapid non-genomic actions of progesterone are implicated in many aspects of female reproduction. Recently, three human homologues of the fish membrane progestin receptor (mPR) have been identified. We combined bioinformatic analysis with expression profiling to define further the role of these mPRs in human reproductive tissues. Sequence analysis confirmed that the mPRs belong to a larger, highly conserved family of proteins, termed 'progestin and adiponectin receptors' (PAQRs). A comparison of the expression of mPR transcripts with that of two related PAQR family members, PAQRIII and PAQRIX, in cycling endometrium and pregnancy tissues revealed markedly divergent expression levels and profiles. For instance, endometrial expression of mPRalpha and gamma and PAQRIX was cycle dependent whereas the onset of parturition was associated with a marked reduction in myometrial mPRalpha and beta transcripts. Interestingly, mPRalpha and PAQRIX were most highly expressed in the placenta, and the tissue expression levels of both genes correlated inversely with that of the nuclear PR. Phylogenetic analysis demonstrated that PAQRIX belongs to the mPR subgroup of proteins. We also validated a polyclonal antibody raised against the carboxy-terminus of human mPRalpha. Immunohistochemical analysis demonstrated more intense immunoreactivity in placental syncytiotrophoblasts than in endometrial glands or stroma. The data suggest important functional roles for mPRalpha, and possibly PAQRIX, in specific reproductive tissues, particularly those that express low levels of nuclear PR. PMID- 16214945 TI - Increased extracellular local levels of estradiol in normal breast in vivo during the luteal phase of the menstrual cycle. AB - Estrogen exposure is a major risk factor for breast cancer. Tissue estrogen originates from the ovaries but a significant portion is also produced by enzyme activity locally in the breast itself. How these enzymes are regulated is not fully understood. The extracellular space, where the metabolic exchange and cell interactions take place, reflects the environment that surrounds the epithelium but there has been no previous study of hormone concentrations in this compartment. In the present study microdialysis was used to measure extracellular estrogen concentrations in breast tissue and abdominal subcutaneous fat in 12 healthy women in vivo. It was found that women with high plasma progesterone levels had significant increased levels of estradiol in breast tissue compared with fat tissue (breast tissue 168+/-6 pM; subcutaneous fat, 154+/-5 pM; P<0.05), whereas women with low plasma progesterone exhibited no difference. Moreover, there was a significant correlation between local breast tissue estradiol and plasma progesterone levels (r=0.709, P<0.01). There was no difference in estrone sulphate in breast and fat tissue regardless of progesterone levels. Estrone was not detectable. The results in this study suggest that progesterone may be one regulator in the local conversion of estrogen precursors into potent estradiol in normal breast tissue. PMID- 16214946 TI - The sex difference of plasma homovanillic acid is unaffected by cross-sex hormone administration in transsexual subjects. AB - There is a close relationship between the brain and the endocrine system. The brain expresses receptors for sex steroids and is capable of metabolizing these hormones. We explored (1) sex differences in homovanillic acid (HVA), a metabolite of the neurotransmitter dopamine, and (2) the effects of cross-sex steroid administration in transsexual subjects. First, we compared plasma HVA levels between 38 male and 34 female healthy volunteers (not using hormone replacement therapy) of a mean age of 72 years (range 65-84 years). Secondly, we measured plasma HVA levels in 15 male-to-female transsexuals treated with 100 microg ethinyl estradiol/day and 100 mg cyproterone acetate/day for 4 months, and in 17 female-to-male transsexuals treated with testosterone esters (250 mg/2 weeks i.m. for 4 months). Plasma HVA levels were lower in elderly men than in elderly postmenopausal women (geometric mean 25.4 nmol/l (percentile (P)10 4.9; P90 69.8) vs 39.0 nmol/l (19.0; 76.1); P=0.027). In transsexuals before cross-sex hormone administration, genetic males also had lower plasma levels of HVA than genetic females (geometric mean 14.8 nmol/l (P10 7.0; P90 35.0) vs 34.3 nmol/l (21.8; 61.4); P<0.001). Cross-sex hormone administration did not affect plasma HVA in either group (P>0.5). The pretreatment sex difference in plasma HVA was unaffected after 4 months of cross-sex hormone administration (P=0.003). The sex difference in plasma HVA was not reversed by cross-sex hormone administration in transsexuals, and was also preserved in elderly subjects. This indicated that differences in dopamine gene expression were largely unaffected by exposure to sex hormone levels in adulthood, but must rather be explained by a sex difference in genetic factors or by the organizing effects of sex hormones during early development. PMID- 16214947 TI - Influence of long-term dietary administration of procymidone, a fungicide with anti-androgenic effects, or the phytoestrogen genistein to rats on the pituitary gonadal axis and Leydig cell steroidogenesis. AB - Procymidone is a fungicide with anti-androgenic properties, widely used to protect fruits from fungal infection. Thereby it contaminates fruit products prepared for human consumption. Genistein-containing soy products are increasingly used as food additives with health-promoting properties. Therefore we examined the effects of long-term dietary administration (3 months) of the anti-androgen procymidone (26.4 mg/animal per day) or the phytoestrogen genistein (21.1 mg/animal per day) to rats on the pituitary-gonadal axis in vivo, as well as on Leydig cell steroidogenesis and on spermatogenesis ex vivo. The procymidone containing diet elevated serum levels of LH and testosterone and, furthermore, Leydig cells isolated from procymidone-treated animals displayed an enhanced capacity for producing testosterone in response to stimulation by hCG or dibutyryl cAMP, as well as elevated expression of steroidogenic acute regulatory protein (StAR), cytochrome P450 side-chain cleavage (P450 scc) and cytochrome P450 17alpha (P450c17). In contrast, the rate of DNA synthesis during stages VIII and IX of spermatogenesis in segments of seminiferous tubules isolated from genistein-treated rats was decreased without accompanying changes in the serum level of either LH or testosterone. Nonetheless, genistein did suppress the ex vivo steroidogenic response of Leydig cells to hCG or dibutyryl cAMP by down regulating their expression of P450 scc. Considered together, our present findings demonstrate that long-term dietary administration of procymidone or genistein to rats exerts different effects on the pituitary-gonadal axis in vivo and on Leydig cell steroidogenesis ex vivo. Possibly as a result of disruption of hormonal feedback control due to its anti-androgenic action, procymidone activates this endocrine axis, thereby causing hyper-gonadotropic activation of testicular steroidogenesis. In contrast, genistein influences spermatogenesis and significantly inhibits Leydig cell steroidogenesis ex vivo without altering the serum level of either LH or testosterone. PMID- 16214948 TI - Regulation of activin A and inhibin B secretion by inflammatory mediators in adult rat Sertoli cell cultures. AB - The regulation of Sertoli cell activin A and inhibin B secretion during inflammation was investigated in vitro. Adult rat Sertoli cells were incubated with the inflammatory mediators, lipopolysaccharide (LPS), interleukin-1beta (IL 1beta), IL-6 and the IL-1 receptor antagonist (IL-1ra) over 48 h in culture. Activin A, inhibin B and IL-1alpha were measured in the culture medium by specific two-site ELISAs. Both IL-1beta- and LPS-stimulated activin A and inhibited inhibin B secretion. LPS also stimulated the production of IL-1alpha in the cultures. In contrast to IL-1beta, IL-6 had no effect on activin A, although it did have a significant inhibitory effect on inhibin B secretion. Ovine follicle-stimulating hormone (FSH) and the cAMP analogue dibutyryl cAMP opposed the actions of IL-1 and LPS by suppressing activin A and IL-1alpha secretion and by stimulating inhibin B. Blocking IL-1 activity in the cultures by addition of an excess of IL-1ra completely prevented the response of activin A to exogenous IL-1beta, and reduced the response to LPS by 50%. In the presence of IL-1ra, basal secretion of inhibin B was increased, but IL-1ra was unable to reverse the suppression of inhibin B by LPS. These data indicate the importance of both IL-1 isoforms in regulating secretion of activin A and inhibin B by mature Sertoli cells during inflammation. The data also establish that inflammation exerts its effects on activin A and inhibin B secretion via other pathways in addition to those mediated by IL-1, and that hormonal stimulation by FSH and cAMP moderates the Sertoli cell response to inflammation. Interference with the complex interactions between these cytokines and hormones may contribute to the disruption of reproductive function that can accompany infection and illness in men. PMID- 16214949 TI - Fibroblast growth factor (FGF) 2 and FGF9 mediate mesenchymal-epithelial interactions of peritubular and Sertoli cells in the rat testis. AB - The role of fibroblast growth factor (FGF) 2 and FGF9 as mediators of cell-cell interactions between Sertoli cells (SCs) and peritubular cells (PCs) was investigated. Using RT-PCR, we demonstrated that SCs and PCs recovered from 20 day-old rats expressed several of the seven FGF receptors (FGFRs), and more specifically the FGFR1 IIIc. FGF2 and FGF9 did not elicit any morphological changes in primary cultures of SCs, nor did they alter the number of SCs in culture. By contrast, changes in shape were observed in FGF2- and FGF9-treated PCs. In addition, FGF2 but not FGF9 enhanced significantly and dose-dependently the number of PCs in culture, indicating that FGF2 was a survival factor for these cells. It was also mitogenic because it enhanced the [3H]thymidine labeling index in PCs. We next examined the effects of FGF2 and FGF9 in a coculture system using 20-day-old rat SCs and PCs, and in an organotypic culture system using XY rat embryonic gonads. In both models, FGF2 and FGF9 were found to promote cellular interactions as evidenced by the extent of cellular reorganization in the coculture system, and cord morphogenesis and growth in the organotypic culture system. A key feature in SC-PC interactions is the synthesis and remodeling of the basement membrane which is co-elaborated by the two cell types. Since basement membrane homeostasis depends upon the coordinated activity of proteinases and inhibitors, the proteinases and inhibitors produced by PCs and SCs degrading or opposing degradation of the major components of the basement membrane were further studied. Specifically, we monitored the metalloproteinases (MMP)-2 and -9 and the tissue inhibitors -1, -2 and -3, the plasminogen activators (PAs) and the PA inhibitor-1, using zymography for the proteinases and Western blots for the cognate inhibitors. Cocultures received FGF or an analog of cAMP in order to prevent cellular reorganization. We found that FGF2 was unique in inducing MMP-9 in coculture. Also, the enhanced levels of the PA inhibitor-1 and the 30 kDa band glycosylated form of tissue inhibitor-3 correlated with the enhanced SC-PC reorganization. It was concluded that FGF2 and FGF9 are morphogens for the formation of testicular cords. PMID- 16214950 TI - Distribution of extracellular signal-regulated protein kinases 1 and 2 in the rat adrenal and their activation by angiotensin II. AB - The adrenal gland of the rat is continuously regenerated through proliferation of a stem cell population in the outer part of the gland. To clarify the location of proliferative events within the adrenal gland, and the factors that stimulate them, rat adrenal capsule preparations, consisting of capsule, zona glomerulosa (ZG) and the outer zona fasciculata (ZF) were maintained in vitro under different conditions of stimulation, for varying periods. Sites of proliferation were identified by 5-bromo-2'-deoxy-uridine (BrdU) staining, and the distribution of classical MAP kinase (MAPK) family members, extracellular signal-regulated kinase (ERK) 1 and 2, immunoreactivity was determined using immunocytochemistry. BrdU staining was limited to the outer glomerulosa and the capsule, where it was enhanced by angiotensin II, but not by a high potassium ion concentration nor by ACTH. In contrast, ERK1/2 immunoreactivity was distributed throughout the ZG and in the medulla, with none detectable in the ZF and reticularis. Furthermore, angiotensin II, potassium ions and ACTH were all shown to induce ERK1 and ERK2 phosphorylation in the ZG. Treatment of adrenal capsule tissue with the specific MAPK kinase inhibitor PD98059 revealed inhibition of ERK1/2 phosphorylation, but no effect on angiotensin II-induced aldosterone secretion. Although the distribution and activation of the MAPK pathway suggest a link with proliferation, the findings clearly designated only the outer part of the glomerulosa and capsule as a potential stem cell population. Further functions should be sought for the apparently silent major part of the glomerulosa. PMID- 16214952 TI - Notice of inadvertent duplicate publication: statin-induced apoptosis of vascular endothelial cells is blocked by dexamethasone. PMID- 16214951 TI - Nuclear factor-kappaB and advanced glycation end-products expression in lacrimal glands of aging rats. AB - Advanced glycation end products (AGEs) increase with aging and induce signaling alterations that lead to inflammation and dysfunction in several tissues. Aging reduces function and insulin signaling in lacrimal glands (LGs). To evaluate whether AGE signaling and insulin secretion in LGs are altered in aging, 24- and 2-month-old male Wistar rats were compared. Immunohistochemistry with confocal microscopy was used to evaluate AGE, AGE receptor (RAGE) and nuclear factor kappaB (NF-kappaB) expression in LGs. Basal tear secretion volume, insulin, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) levels in tears and LGs and peroxidase activity in LG tissue were measured. Insulin secretion from isolated LGs and pancreatic beta-cells was compared in the supernatant of aging and control rats in vitro by RIA after stimulation with 2.8 16.7 mM glucose, carbachol and KCl. AGE, RAGE and NF-kappaB expression was higher in LGs of aging compared with young rats. Basal tear secretion and peroxidase activity were significantly lower in the aging group (P=0.016 for both assays). IL-1beta and TNF-alpha levels were higher in tears of aging rats compared with young rats (P=0.007 and 0.05 respectively); however, even though aging rats were insulin-resistant (as confirmed by the insulin-tolerance test), the insulin levels in the tear film of aging and control rats were similar in vivo and in vitro. The higher expression of AGEs, RAGE and NF-kappaB in LGs of aging rats is accompanied by systemic insulin resistance and may be involved in LG and tear film alterations but does not affect insulin secretion in the tear film. These observations indicate that metabolic events may be related to LG and tear film dysfunctions in aging. PMID- 16214953 TI - Activation of extracellular signal-regulated kinase signaling by epidermal growth factor mediates c-Jun activation and p300 recruitment in keratin 16 gene expression. AB - In studies of gene regulation of keratin 16, we reported previously that simian virus 40 promoter factor 1 shows a functional cooperation with c-Jun and coactivators p300/CBP in driving the transcriptional regulation of epidermal growth factor (EGF)-induced keratin 16 gene expression. In the present study, we found that the stimulated expression of keratin 16 by EGF was mediated mainly through the mitogen-activated protein kinase kinase-extracellular signal regulated kinases 1 and 2 (ERK1/2) signaling pathway. Ser63 and Ser73 on the c Jun NH(2)-terminal transactivation domain could be phosphorylated in cells treated with EGF; nevertheless, we found that the c-Jun COOH terminus played a pivotal role in EGF-induced expression of keratin 16. The activation of keratin 16 by EGF treatment could not be enhanced by the overexpression of myc-c-JunK3R, in which three putative acetylation lysine residues on the c-Jun COOH terminus were all mutated into arginines, suggesting that c-Jun acetylation on the COOH terminus might partially play a functional role in this system. In addition, by using a chromatin immunoprecipitation assay and a DNA affinity precipitation assay, EGF treatment up-regulated the p300 recruitment through ERK signaling to the promoter region in regulating keratin 16 transcriptional activity. Furthermore, the enhancement of acetyl-histone H3 to the keratin 16 chromatin promoter induced by EGF was also mediated via ERK activation. In conclusion, these results strongly suggest that both c-Jun induction and p300 recruitment to gene promoter, mediated through ERK activation, played an essential role in regulating keratin 16 gene expression by EGF. p300 mediated and regulated EGF induced keratin 16 gene expression, at least in part, through multiple mechanisms, including a selective acetylation of c-Jun and histone H3. PMID- 16214954 TI - Aryl hydrocarbon receptor regulates distinct dioxin-dependent and dioxin independent gene batteries. AB - Conventional biochemical and molecular techniques identified previously several genes whose expression is regulated by the aryl hydrocarbon receptor (AHR). We sought to map the complete spectrum of AHR-dependent genes in male adult liver using expression arrays to contrast mRNA profiles in Ahr-null mice (Ahr(-/-)) with those in mice with wild-type AHR (Ahr(+)(/)(+)). Transcript profiles were determined both in untreated mice and in mice treated 19 h earlier with 1000 microg/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression of 456 ProbeSets was significantly altered by TCDD in an AHR-dependent manner, including members of the classic AHRE-I gene battery, such as Cyp1a1, Cyp1a2, Cyp1b1, and Nqo1. In the absence of exogenous ligand, AHR status alone affected expression of 392 ProbeSets, suggesting that the AHR has multiple functions in normal physiology. In Ahr(-/-) mice, only 32 ProbeSets exhibited responses to TCDD, indicating that the AHR is required for virtually all transcriptional responses to dioxin exposure in liver. The flavin-containing monooxygenases, Fmo2 and Fmo3, considered previously to be uninducible, were highly induced by TCDD in an AHR dependent manner. The estrogen receptor alpha as well as two estrogen-receptor related genes (alpha and gamma) exhibit AHR-dependent expression, thereby extending cross-talk opportunities between the intensively studied AHR and estrogen receptor pathways. p53 binding sites are over-represented in genes down regulated by TCDD, suggesting that TCDD inhibits p53 transcriptional activity. Overall, our study identifies a wide range of genes that depend on the AHR, either for constitutive expression or for response to TCDD. PMID- 16214955 TI - Aryl hydrocarbon receptor activation inhibits regenerative growth. AB - There is considerable literature supporting the conclusion that inappropriate activation of the aryl hydrocarbon receptor (AHR) alters cellular signaling. We have established previously that fin regeneration is specifically inhibited by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in adult zebrafish and have used this in vivo endpoint to evaluate interactions between AHR and growth-controlling pathways. Because there are experimental limitations in studying regeneration in adult animals, we have developed a larval model to evaluate the effect of AHR activation on tissue regeneration. Two-day-old zebrafish regenerate their amputated caudal fins within 3 days. Here, we demonstrate that TCDD specifically blocks regenerative growth in larvae. The AHR pathway in zebrafish is considerably more complex than in mammals, with at least three zebrafish AHR genes (zfAHR1a, zfAHR1b, and zfAHR2) and two ARNT genes (zfARNT1 and zfARNT2). Although it was presumed that the block in regeneration was mediated by AHR activation, it had not been experimentally demonstrated. Using antisense morpholinos and mutant fish lines, we report that zfAHR2 and zfARNT1 are the in vivo dimerization partners that are required for inhibition of regeneration by TCDD. Several pathways including fibroblast growth factor (FGF) signaling are essential for fin regeneration. Even though impaired FGF signaling and TCDD exposure both inhibit fin regeneration, their morphometric response is distinct, suggesting that the mechanisms of impairment are different. With the plethora of molecular and genetic techniques that can be applied to larval-stage embryos, this in vivo regeneration system can be further exploited to understand cross talk between AHR and other signaling pathways. PMID- 16214957 TI - 2-aminoethoxydiphenyl borate as a prototype drug for a group of structurally related calcium channel blockers in human platelets. AB - We have synthesized a series of 2-aminoethoxydiphenyl borate (2-APB, 2,2-diphenyl 1,3,2-oxazaborolidine) analogs and tested their ability to inhibit thrombin induced Ca(2+) influx in human platelets. The analogs were either synthesized by adding various substituents to the oxazaborolidine ring (methyl, dimethyl, tert butyl, phenyl, methyl phenyl, and pyridyl) or increasing the size of the oxazaborolidine ring to seven- and nine-membered rings. NMR analysis of the boron containing analogs suggests that each of them exist as a ring structure through the formation of an N-->B coordinate bond (except for the hexyl analog). The possibility that these boron-containing compounds formed dimers was also considered. All compounds dose-dependently inhibited thrombin-induced Ca(2+) influx in human platelets, with the 2,2-diphenyl-1,3,2-oxazaborolidine-5-one derivative having the weakest activity at 100 microM, whereas the (S)-4-benzyl and (R)-4-benzyl derivatives of 2-APB were approximately 10 times more potent than the parent 2-APB. Two nonboron analogs (3-methyl and 3-tert-butyl 2,2 diphenyl-1,3-oxazolidine) were synthesized; they had approximately the same activity as 2-APB, and this implies that the presence of boron was not necessary for inhibitory activity. All of the compounds tested were also able to inhibit thrombin-induced calcium release. We concluded that extensive modifications of the oxazaborolidine ring in 2-APB can be made, and Ca(2+)-blocking activity was maintained. PMID- 16214956 TI - Revisiting the postulated "unitary glutamate receptor": electrophysiological and pharmacological analysis in two heterologous expression systems fails to detect evidence for its existence. AB - Several years ago evidence for a so-called "unitary glutamate receptor" was published. This unique type of glutamate receptor was reported to be activated by the traditional agonists of all three major glutamate receptor subfamilies [i.e., alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA), kainate, and N methyl-d-aspartate (NMDA)] in a glycine-dependent as well as magnesium-blockable manner and was reported to consist of an NR1 subunit coexpressed with the kainate binding protein (KBP) from Xenopus laevis, XenU1. To re-examine the existence of such a receptor, we cloned two splice variants of the X. laevis NMDA receptor subunit NR1, XenNR1-4a and XenNR1-4b, and expressed them in X. laevis oocytes as well as in human embryonic kidney (HEK) 293 cells, either alone or with the X. laevis KBP subunit XenU1. In addition, we coexpressed XenU1 separately with all eight splice variants of the rat NR1 subunit. In no case did we see evidence of a unitary glutamate receptor pharmacology. In HEK293 cells, we did not get receptor response unless an NR2 subunit was coexpressed. In X. laevis oocytes, we did observe responses to glutamate/glycine as well as small responses to glycine alone, but these were independent of coexpressed XenU1. Neither AMPA nor kainate ever elicited significant responses. Because we verified that XenU1 is expressed and inserted into the plasma membrane of HEK293 cells, we conclude that XenU1 and NR1 do not form the postulated unitary glutamate receptor. Furthermore, successful amplification of a fragment of a X. laevis NR2 subunit indicates that X. laevis uses NR2 subunits and not XenU1 to form heteromeric complexes with NR1. PMID- 16214958 TI - Thoughts on the law. PMID- 16214959 TI - Antioxidants: bridging the gap between biochemistry and clinical practice. PMID- 16214960 TI - The biochemistry of antioxidants revisited. AB - Biochemical relationships between oxidative stress, antioxidant nutrients, and chronic diseases are complicated and often conflicting. Basic research supports the concept that reactive oxygen species precipitate changes that result in oxidative damage to lipid, protein, and DNA biomolecules. Oxidative stress is implicated in the development of cancer, cardiovascular disease, diabetes, sepsis, various eye diseases, and neurologic conditions. Supplementation with antioxidant nutrients seems plausible to counter the effects of oxidative stress, but the preferred mode of delivery for these nutrients may be through the patient's diet rather than as supplements to the diet. In fact, evidence supporting consumption of at least 5 servings of fruits and vegetables continues to grow. To better understand the role of antioxidant nutrients in disease promotion or prevention, this review will discuss basic nutritional biochemistry relating to oxidative stress and antioxidant defense systems, followed by a discussion of the metabolism (vitamins E, C, A) and interrelationships of select antioxidant nutrients. PMID- 16214961 TI - Energy expenditure in the ICU. PMID- 16214962 TI - Methods of assessing energy expenditure in the intensive care unit. AB - An essential component in developing the nutrition support plan for hospitalized patients is evaluating energy requirements. Assessing energy expenditure (EE) and identifying requirements in the critically ill patient present the clinician with a challenge; how to prevent overfeeding and minimize underfeeding? Both under- and overfeeding have been associated with increased morbidity and mortality. It is known that critical illness alters EE. This alteration is hormonally mediated and is characterized by changes in metabolic processes. Methods used by clinicians to assess EE in the critically ill patient vary significantly. It is the purpose of this review to outline the various methods for evaluating EE in critical illness with emphasis on their benefits and limitations. PMID- 16214963 TI - Improved equations for predicting energy expenditure in patients: the Ireton Jones Equations. PMID- 16214964 TI - Management and knowledge of enteral nutrition in intensive care units in a city in Belgium. AB - The administration of enteral nutrition in intensive care unit (ICU) patients is largely managed by nurses. However, the degree of knowledge, interest, and training in this field can differ considerably among nurses and among ICUs. Such differences may lead to variations in the way in which enteral nutrition is used. To investigate these issues, a questionnaire survey was sent to the nursing staffs of 5 ICUs in Belgium. The response rate was 68%. Although theoretical knowledge of enteral nutrition was globally poor, its advantages over parenteral nutrition were usually known. Responses to questions related to practical issues associated with enteral feeding showed more institution-specific answers than interindividual differences. PMID- 16214965 TI - Energy requirements in critically ill patients: how close are our estimates? AB - A retrospective analysis of 55 mechanically ventilated critically ill patients was conducted to determine adequacy of nutritional support (total parenteral nutrition or enteral nutrition) according to requirements established by indirect calorimetry. Patients who received 90% to 110% of the established energy requirements as measured by indirect calorimetry were defined as adequately fed. At the time of the indirect calorimetry measurements, all patients were receiving their targeted nutritional support, as assessed by the unit dietitian, who used predictive formulas to assess patients. Indirect calorimetry results showed that 25% of the patients were overfed (receiving >110% of energy requirements), 35% were underfed (receiving <90% of energy requirements), and 40% were adequately fed (receiving 90% to 110% of energy requirements). We determined that critically ill patients with a body mass index <20 kg/m2 were the most likely group to be assessed inappropriately by available regression equations. If indirect calorimetry measurement is unavailable, we suggest using an empiric formula of 37 kcal/kg for critically ill patients with a body mass index <20 kg/m2. PMID- 16214966 TI - The loss of innocence. PMID- 16214967 TI - Venous access: "What's it to you?". PMID- 16214968 TI - Multivitamins for the millennium. PMID- 16214969 TI - The ins and outs of venous access: part I. AB - Selection of the proper venous access device is important to maximize patient benefit and minimize patient discomfort, morbidity, mortality, and cost. The decision of which device to use is based on whether or not the patient requires central venous access and whether the need is short-term (<6 to 8 weeks) or long term. Short-term venous access devices include short peripheral IV catheters, midline catheters, peripherally inserted central catheters (PICC), and central venous catheters (CVC). This article reviews each of these short-term devices and their indications, contraindications, advantages, and disadvantages. Part 1 covers Venous Anatomy and Short-Term Venous Access; Part 2, to be published in the June issue, covers Long-Term Venous Access. PMID- 16214970 TI - The peripherally inserted central catheter: what are the current indications for its use? AB - The use of peripherally inserted central catheters (PICCs) has dramatically increased since first being used for IV therapy in the 1980s. Currently, sales of PICCs are second only to acute care central venous catheters and exceed all other types of long-term venous access devices. The use of PICCs has also increased dramatically in the home care setting. A number of studies have examined the incidence of PICC complications in comparison to other central venous access devices. Although complications differ, most reports have concluded that the type and rate of PICC complications compare favorably with other access devices in the short-term; however, dwell times beyond a few weeks have been associated with decreased complication-free days and lower device survivability rates. The PICC seems to be most appropriate for acute care or short-term home care use and may not be the ideal central venous access device when the need for long-term access is anticipated. PMID- 16214971 TI - Routine chest radiography following imaging-guided placement of tunneled central lines: a waste of time, money, and radiation. AB - BACKGROUND: This study sought to evaluate routine chest radiography following placement of tunneled central lines using combined ultrasound and fluoroscopic guidance. MATERIALS AND METHODS: A prospective study of 150 consecutive patients who underwent placement of tunneled central lines in the vascular radiology suite. Ultrasound-guided vein puncture was performed with an 18-gauge needle in each case, and the access site was noted. Line position was confirmed by fluoroscopy. Following the procedure, 50 patients had both an on-table digital chest radiograph and a conventional chest radiograph. Subsequent patients had a digital radiograph and a fluoroscopic image grab. Final line tip position was scored, and complications were recorded. RESULTS: Line placement was optimal (95%) or acceptable (5%) in all patients. Line tip position could be satisfactorily evaluated on supine fluoroscopy. Mean fluoroscopic x-ray dose was 0.5 cGy/cm2. Digital chest x-ray dose was 9.0 cGy/cm2, and formal chest radiography dose was 12.0 cGy/cm2. The only complications were 2 carotid artery punctures without clinical sequelae. CONCLUSION: When lines are placed under imaging guidance with ultrasound to direct the venous puncture, complications are rare and are not likely to be clinically important. Conventional and digital chest radiographs do not contribute clinically relevant information but do add to the radiation dose, time, and expense of the procedure. PMID- 16214972 TI - Short bowel syndrome in the elderly. AB - BACKGROUND: The need for massive intestinal resection in the elderly presents an ethical dilemma. The mortality rate for elderly patients who undergo this procedure is high, and quality of life with the short bowel syndrome (SBS) in this group is often questioned. This study evaluated the clinical outcome of patients >70 years of age who underwent intestinal resection leading to SBS. METHODS: This was a retrospective study of 23 patients ranging in age from 70 to 92 years, who were evaluated over a 20-year period. RESULTS: There were 11 (48%) early postoperative deaths. Parenteral nutrition (PN) was required for 8 of 12 discharged patients. Five (63%) of the 8 remained on PN permanently, and 3 were transitioned to enteral nutrition (EN). Three patients (PN = 2, EN = 1) died within a year of resection. Four (67%) of 6 patients surviving 5 years received EN only. Five of 10 patients with remnant length <120 cm were discharged, all on PN. PN was later discontinued in 2 patients. Seven (54%) of 13 patients with remnants > 120 cm were discharged; 3 of the 7 remained on PN. Resection was performed for mesenteric vascular disease (n = 13), radiation or cancer (n = 6), and other benign conditions (n = 4). Underlying diagnosis did not influence mortality rate or need for PN. Intestinal continuity was restored in 3 of the 8 patients discharged with an ostomy. Stricturoplasty was performed in 2 patients to improve intestinal function. CONCLUSIONS: Massive resection in the elderly has a high early mortality rate. Long-term survival is achieved in 50% of discharged patients, especially if they can transition to full EN. Nutritional prognosis is influenced by remnant length more than by underlying disease. Despite their advanced age, these patients are often candidates for further surgical procedures to improve intestinal function. PMID- 16214973 TI - Enteral formula use in children after small bowel transplant. AB - The ultimate goal of intestinal transplantation (ITx) is the maintenance of nutritional status enterally/orally. We retrospectively identified children who had received ITx since the inception of our Intestinal Care Center in December 1996 (n = 24; median age, 2.6 years). Two patients died within 2 months of transplant. Enteral formulas used in the remaining 22 patients included the following: amino acid, trace long chain fatty acids (LCT; n = 4); amino acid, medium chain fatty acids (MCT)/LCT fat mix (n = 13); amino acid, LCT fat (n = 1); and peptide, MCT/LCT fat mix (n = 3). Feedings were initiated on an average of 13 days after ITx. The median number of days to complete the total parenteral nutrition (TPN) wean was 30 days, and stoma output measured on an average of 37 mL/kg per day at 1 month posttransplant. Nine patients (41%) advanced to oral intake alone within 2 to 30 months, and 5 patients (23%) were diagnosed with milk allergy through the RAST test (Pharmacia, Uppsala, Sweden). No differences in the initiation of therapy, advancement to oral intake, stoma output, or TPN weaning were observed by the type of formula used. Using z-score statistics, positive linear growth was achieved in 7 of 21 children (33%) over the 1-year period, whereas linear growth velocity was maintained in an additional 6 patients (29%). Successful advancement to total enteral/oral intake and positive growth after ITx can be achieved with either an amino acid- or peptide-based, partial MCT enteral formula initiated within 2 weeks of transplant. Monitoring for posttransplant allergy is recommended because of the high rate of postoperative allergy symptomology. PMID- 16214974 TI - Comparison of continuous vs intermittent nasogastric enteral feeding in trauma patients: perceptions and practice. AB - Enteral nutrition support (ENS) may be administered by continuous administration or by intermittent bolus, but few studies have compared the advantages and limitations associated with these methods in the trauma patient population. The purpose of this prospective randomized study was to evaluate the impact of continuous vs intermittent nasogastric enteral feeding on gastrointestinal tolerance, pulmonary aspiration, and nutritional indices. A survey of medical and surgical intensive care unit nursing staff was performed to assess perceptions and attitudes toward these 2 delivery methods. Eighteen trauma patients [Injury Severity Score (ISS) > or = 20] were enrolled in the study; 9 received continuous ENS (CENS) and 9 received intermittent bolus ENS (IENS). Interruption of ENS delivery occurred in more IENS than CENS patients due to elevated residuals and emesis. Diarrhea occurred in more patients (5/9 vs 2/9) and for a longer duration (14/65 vs 6/49 ENS days) in the IENS group compared with the CENS group. Aspiration was detected in 1 IENS patient. Method of nutrient delivery did not seem to influence urine urea nitrogen (UUN) measurements or prealbumin concentrations. Nurses surveyed in the study (n = 25) preferred CENS (84%) compared with IENS (12%). Data from this study suggest that CENS through a nasoenteric feeding tube may facilitate nutrient intake with less gastrointestinal complications in severely injured trauma patients compared with IENS. In addition, the majority of nurses surveyed preferred the continuous method for nutrient delivery. PMID- 16214975 TI - Early initiation of enteral feeding after percutaneous endoscopic gastrostomy tube placement. AB - Enteral feeding through the percutaneous endoscopic gastrostomy (PEG) tube is usually initiated about 12 to 24 hours after insertion of the tube. There have been earlier studies evaluating the efficacy of early initiation of enteral feedings that had encouraging results. However, delayed initiation of feeding following PEG placement continues to be practiced widely. We believe that feeding can be done earlier without any increase in associated morbidity or mortality and with obvious reduction in the need for parenteral nutrition and healthcare costs. We evaluated a protocol to initiate enteral nutrition 4 hours after the PEG tube insertion with subsequent discharge of the outpatients on the same day. We conducted a prospective study to assess the efficacy of early initiation of PEG feeding. We enrolled 77 patients in our study who were having PEG tubes placed for enteral feeding. Only patients who had a PEG placed for gastric venting procedures were excluded from our study. During the course of our study, no patient had to be excluded for the latter reason. Patients were evaluated by the physician performing the procedure, 4 hours after the tube was inserted. Their vital signs were checked, and a thorough abdominal examination was performed. Minimal tenderness around the PEG site was the most frequent finding. Otherwise, all the patients had a benign abdominal examination. The tube was flushed with 60 mL of sterile water. Following the examination, orders were given to restart the feedings. These patients were followed for a 30-day period to evaluate complications associated with PEG tube placement and early initiation of PEG feeding. There was one case of aspiration pneumonia (1.3%) and one death that was attributed to the underlying disease out of our 77 patients. Early initiation of enteral feeding after PEG tube placement can be successfully completed with a systematic protocol and close observation. Not only was this protocol found to be safe, it can also have significant cost savings by eliminating the need for inpatient hospitalization for the procedure. PMID- 16214976 TI - Cloning kitties. PMID- 16214977 TI - Can we justify continued interest in indirect calorimetry? PMID- 16214978 TI - Pause for thought: enteral nutrition in the critically ill. PMID- 16214979 TI - Preventing enteral tube feeding-associated aspiration: something no one should get blue in the face over. PMID- 16214980 TI - The ins and outs of venous access: part II. PMID- 16214981 TI - Is there a role for specialized enteral nutrition in the intensive care unit? AB - The enteral route is preferred for nutrition support of critically ill patients who have a functional gastrointestinal tract. A variety of specialized formulas are available to help manage the complications often seen in these patients. This review summarizes the literature supporting the use of formulas marketed for liver disease, wound healing, immune-system enhancement, pulmonary and renal disease, and diabetes mellitus. With the exception of the immune-enhancing formulas, convincing evidence for the use of these formulas in critically ill patients is lacking. Further studies are warranted. PMID- 16214982 TI - Food dye use in enteral feedings: a review and a call for a moratorium. AB - Pulmonary aspiration of gastric contents is common in enterally fed patients. Tinting enteral feedings with blue dye is thought to aid the early detection of aspiration in hospitalized patients. The blue-dye method is popular despite evidence that it is not sensitive. Reports of absorption of blue dye from enteral feedings in patients with sepsis and other critical illnesses are increasing. The presence of blue and green skin and urine, and serum discoloration has been linked with death. FD&C Blue No.1 and related dyes have toxic effects on mitochondria, suggesting that dye absorption is harmful. This study reviews the literature on the dye method and dye pharmacology, reports the results of a survey of current dye use, and describes 2 recent deaths associated with blue-dye absorption. We concluded that the use of blue dye in enteral feedings should be abandoned and replaced by evidence-based methods for the prevention of aspiration. PMID- 16214983 TI - Pitfalls in predicting resting energy requirements in critically ill children: a comparison of predictive methods to indirect calorimetry. AB - BACKGROUND: Critical illness in children is thought to have profound effects on nutritional status. It is essential to avoid complications associated with inadequate nutrition support and delivery of excess energy. OBJECTIVE: To compare the results of several commonly used methods for predicting energy requirements in a group of critically ill children indirect calorimetry was used to measure energy expenditure in these children. DESIGN: Resting energy expenditures estimated by different prediction methods for energy were compared with measurements of actual resting energy expenditure obtained by indirect calorimetry in 52 children admitted to a pediatric intensive care unit. Agreement between each predictive method and indirect calorimetry was evaluated by Bland Altman limits of agreement and by whether the methods met the predetermined criterion for accuracy of within 10% of the measured value. RESULTS: None of the equations predicted individual values accurately. Each of the predictive equations gave a wide and variable scatter of predicted values around the median. The recommended dietary allowance for energy was the least accurate and differed significantly even from the other predictive methods, overestimating energy expenditure in 50 of 52 patients. None of the remaining methods stood out as being more precise. CONCLUSIONS: Predictive methods commonly used to estimate energy expenditure in critically ill children are very imprecise and may lead to overprovision or underprovision of nutrition support. Resting energy expenditure should be measured by indirect calorimetry whenever possible. PMID- 16214984 TI - Considerations in feeding obese patients: a review of a classic article. 1986. PMID- 16214985 TI - Central venous catheterization (CVC) has become increasingly common in the management of critically ill patients. PMID- 16214986 TI - Adjuvant perioperative nutrition support in cancer patients. PMID- 16214987 TI - The Malpractice Crisis of 2002. PMID- 16214988 TI - The pathophysiology and treatment of cancer cachexia. AB - Cachexia in cancer patients is a major cause of morbidity and mortality. The syndrome of cachexia seems to be a facet of the inflammatory response to injury. This proinflammatory state is driven by an interdependent web of mediators such as cytokines, neurotransmitters, prostaglandins, neuroendocrine hormones, and tumor-derived catabolic factors. The resulting metabolic response to cancer prevents the effective use of calories provided and therefore blocks anabolism. The most promising interventions in cancer cachexia seem to depend on modulation of the inflammatory state (including nonsteroidal anti-inflammatory drugs and fish oil-based preparations). Suppression of the inflammatory milieu seems to allow nutrition to have an anabolic effect, resulting in gains in lean tissue with the potential to improve patient survival and quality of life. PMID- 16214989 TI - Nutritional therapy for cancer-induced weight loss. AB - Nutrition issues facing people with cancer are significant. The role of nutrition therapy is often assumed to be less important than pharmacologic interventions as outcomes are less clear in the literature. Especially important has been the focus on traditional outcomes such as morbidity and mortality rather than adopting a more patient-centered approach and including quality of life as a key outcome. There is a general failure to recognize weight loss early enough and to implement nutrition interventions, which are effective. The nutrition intervention should be composed of both a prescription and an implementation method. To achieve compliance, the nutrition therapy should be implemented in an aggressive manner. Data show that when compliance is achieved, outcomes in the form of improved quality of life and functional capacity can be achieved. For people with cancer and their care-givers, outcomes such as quality of life have meaning, and for patients with poor prognoses, this may hold more meaning than mortality and morbidity outcomes. Clinicians need to implement triage systems to identify patients for early intervention and focus on nutrient combinations. They need to be confident that with the right prescription and implementation, nutrition therapy can be successful and bring real benefit to patients with cancer. PMID- 16214990 TI - Perioperative nutritional support as an adjunct to surgical therapy for cancer. PMID- 16214991 TI - Arginine: a clinical perspective. AB - Dietary arginine is commonly found in both plant and animal products and has also become popular as a dietary supplement over the past 20 years. Arginine has been shown to have a wide variety of effects on the body, including impact on the cardiovascular system, the immune system, and wound healing. Recently it has also been shown to interact with popular pharmaceutical therapies (ie, statin drugs) in a positive way. Research indicates that arginine is safe and has positive effects on the body when used for relatively short periods of time. Data regarding arginine's safety with long-term use in pharmaceutical amounts is still not available for select patient populations (ie, cancer patients). PMID- 16214992 TI - Nutrition support in renal failure. PMID- 16214993 TI - Scurvy in a patient with presumptive oral lichen planus. AB - Scurvy is a disease caused by vitamin C deficiency. In this study, a patient with scurvy is presented with soreness and ulcerations of the mouth that were previously diagnosed as oral lichen planus. This disease is relatively more common than scurvy in the United States. Physicians should be aware that even in developed countries, soreness of the mouth and ulceration can be seen in patients with scurvy and potentially be misdiagnosed as oral lichen planus. PMID- 16214994 TI - Evaluating the effectiveness of a micronutrient assessment tool for long-term total parenteral nutrition patients. AB - Monitoring parameters for patients receiving long-term home parenteral nutrition (HPN) should include assessment of micronutrient status to avoid deficiencies and potential excesses of any micronutrient. This evaluation examined the effectiveness of a standardized micronutrient assessment tool administered semiannually to long-term HPN consumers, for identifying signs/symptoms of a suspected deficiency or excess. All patients assessed were receiving long-term HPN from a single home-infusion provider. Of the 275 micronutrient assessments evaluated, 30 patients were found to have a suspected deficiency. Laboratory tests were ordered for 13 patients: deficiencies were confirmed in 6 patients, no deficiency was found in 4 patients, and 1 patient was not deficient in the suspected micronutrients but was found deficient in others. The micronutrient assessment tool is effective in determining the suspected presence of micronutrient deficiencies. All long-term HPN patients should receive ongoing monitoring by a home care clinician, including an assessment of micronutrient status, to ensure adequacy of the nutrition support regimen. PMID- 16214995 TI - Clinical dilemma in cancer: is tumor growth during nutrition support significant? PMID- 16214996 TI - Intravenous hyperalimentation and cancer: a historical perspective. 1986. PMID- 16214997 TI - Dietary management of patients in chronic renal failure. 1975. PMID- 16214998 TI - Immunonutrition in critical illness. PMID- 16214999 TI - Immunonutrition in the critically ill patient: putting the cart before the horse? PMID- 16215000 TI - Short-term enteral access--efficient or deficient. PMID- 16215001 TI - Ins and outs of enteral access. Part 1: short-term enteral access. AB - Proper selection of the type of enteral access in a given clinical situation is important to maximize the benefit to the patient and minimize the discomfort to the patient, the morbidity and mortality, and the cost. This decision is based on whether or not the patient needs short-term (<4-6 weeks) or long-term enteral nutrition support. This article focuses on short-term access including nasogastric (NG) and nasoenteric (NE) tubes and reviews each of these enteral access tubes along with their indications, contraindications, advantages, and disadvantages. PMID- 16215002 TI - Does lipid hang time make a difference? Time is of the essence. PMID- 16215003 TI - Parent perceptions of mealtime behaviors in children fed enterally. AB - BACKGROUND: Parent reports of mealtime behavioral problems were examined within a sample of medically complicated young children with documented feeding difficulties. Mealtime behavior problems of children who were fed orally were compared with children who received their total nutrition enterally. METHODS: A chart review of 140 pediatric patients presenting to an outpatient Interdisciplinary Feeding Team (IFT) clinic was conducted. The sample consisted of 81 males and 59 females ranging from 1.5 months to 12.6 years of age (mean age, 36.5 months). Information included for analyses was obtained from the IFT intake form, final IFT report, and the Behavioral Pediatrics Feeding Assessment Scale (BPFAS). RESULTS: Parents of children in both groups found child mealtime behaviors equally problematic according to mean intensity ratings and frequency of problems regarding both parent behavior and child behaviors. CONCLUSIONS: Regardless of feeding status (enteral versus oral feeding) of the child, parents of children with feeding and nutrition concerns benefit from intervention and support surrounding mealtimes. The specific mealtime strategies and needs of the parents, however, may differ depending on the feeding status of the child. PMID- 16215004 TI - Weight-based ordering: an evaluation of increased guideline use in hospital total parenteral nutrition dosing. AB - BACKGROUND: In the past, parenteral nutrition in the Calgary Health Region was ordered as volumes of standard solutions, which limited individualization. Ordering total parenteral nutrition (TPN) that falls within macronutrient dosing guidelines may minimize complications associated with TPN, such as hyperglycemia, azotemia, hepatic steatosis, or continued malnutrition and catabolism. The Foothills Medical Centre in Calgary changed to a weight-based ordering system for TPN in 1999. This study's purpose was to determine if this change increased adherence to TPN dosing guidelines. METHODS: Macronutrient doses in TPN solutions ordered as standard solutions were compared with those ordered by weight. Mean protein, dextrose, lipid, and kilocalorie doses and the number of orders deviating from guidelines were examined. RESULTS: Weight-based dosing showed a significant reduction in deviation from guidelines for kilocalorie dose compared with TPN ordered as standard solutions. There also was a significant increase in mean protein dose and reductions in mean dextrose load and mean kilocalorie dose in the weight-based TPN group only, suggesting these changes were caused by the change in ordering method. CONCLUSIONS: Overall, weight-based ordering increased adherence to TPN dosing guidelines. The study did not have the statistical power to show significant differences between weight-based or standard TPN dosing; however, several trends were shown. PMID- 16215005 TI - Persistent left superior vena cava: clinical implications for central venous cannulation. AB - In general, persistence of the left superior vena cava (LSVC), the most common anomaly of the venous circulation, is asymptomatic. Diagnosis of a catheter in a persistent LSVC is not straight forward, and a LSVC can create difficulties during central venous and pulmonary artery catheterization. We discuss the differential diagnosis of left-sided central venous catheters (CVC). Finally a directive is given to prevent dilator-induced vessel injuries. We report the cannulation of an unsuspected persistent LSVC in two patients. A dilator-induced vessel injury contributed significantly to the fatal outcome in the first case. On a plain chest X-ray, a catheter in a LSVC will run down the left mediastinal border and can be confused with other intravascular malpositions and extravascular malpositions. Contrast-enhanced lateral chest radiograph is an inexpensive and readily available method that can be used to determine exact position. PMID- 16215006 TI - Energy expenditure assessment and validation after acute spinal cord injury. AB - BACKGROUND: This study was designed to determine the applicability of the Harris Benedict (HB) equation in predicting energy requirements after acute spinal cord injury (SCI) and to evaluate the accuracy of a 30-minute energy expenditure measurement to determine 24-hour requirements. Prealbumin (PAB) was monitored weekly to assess the patient's response to the nutrition support regimen. METHODS: Patients with acute isolated SCI requiring mechanical ventilation were studied for 4 weeks postinjury. Predicted energy expenditure was estimated using a modified HB equation (HBEE). The protein goal was 2.0 g/kg or 0.23 g nitrogen/kg admission body weight/day. Nutrition support was initiated within 48 hours if medically feasible. Indirect calorimetry was performed weekly to obtain 24-hour measured total energy expenditure (MTEE) and measured resting energy expenditure (MREE) values. Respiratory quotient, PAB levels, urinary urea nitrogen (UUN), and calculated nitrogen balance were evaluated weekly. RESULTS: Eleven tetraplegic men with a mean age of 32 +/- 8 years were studied. Mean MTEE equaled 95% to 100% HBEE. Mean MREE was not significantly different from the mean MTEE. There was a significant correlation between mean predicted and measured 24 hour energy expenditure for each data point. Mean UUN excretion ranged from 22.3 to 28.5 g/d that resulted in a mean negative nitrogen balance for all 4 weeks. PAB improved significantly over the 4-week study period (p < .001). CONCLUSIONS: There was a strong correlation between HBEE and MTEE. A brief REE measurement is adequate to determine daily calorie requirements. Visceral protein synthesis can be achieved despite a negative nitrogen balance. PMID- 16215007 TI - A retrospective review of the course of patients with pancreatitis discharged on jejunal feedings. AB - INTRODUCTION: Although parenteral nutrition (PN) has been the standard nutrition therapy in patients with pancreatitis requiring nutrition support, it is associated with a higher rate of catheter-related sepsis and gut atrophy. Research suggests enteral nutrition (EN) is possible in patients with pancreatitis without exacerbating symptoms when infused jejunally. The purpose of this study was to review the course of patients with resolving pancreatitis discharged to home on EN. METHODS: The medical records of 33 patients with a percutaneous endoscopic gastrostomy tube with jejunal extension (PEG-J) or nasojejunal tube (NJ) who received home EN were reviewed. The data collected included duration of EN, formula used, goal and maximum tolerated flow rates, anthropometric measures, and gastrointestinal complications. RESULTS: Ninety seven percent of patients received a standard polymeric formula providing an average of 1845 +/- 421 kcal/d. Forty-two percent of patients took pancreatic enzyme supplements. Complications occurring in patients included nausea and vomiting (42%), feeding rate intolerance (18%), diarrhea (12%), and PEG site infection (27%). Seventy-seven percent of patients achieved nutritional goals. CONCLUSION: Standard polymeric EN seems to be safe and efficacious in the home setting for patients with resolving pancreatitis. PMID- 16215008 TI - Nutrition therapy in liver disease. PMID- 16215009 TI - Nutritional aspects in liver disease and liver transplantation. PMID- 16215010 TI - Nutrition support in the liver transplant patient. PMID- 16215011 TI - Pediatric intestinal transplantation--review of current practice. PMID- 16215012 TI - Commentary on "Three years clinical experience with intestinal transplantation" and the nutritional implications. 1994. PMID- 16215013 TI - Plasma zinc is an insensitive predictor of zinc status: use of plasma zinc in children with sickle cell disease. AB - BACKGROUND: This study was designed to explore the use of plasma zinc in determining zinc deficiency in children with sickle cell disease-SS (SCD-SS) as indicated by a growth response to zinc supplementation. METHODS: Fasting plasma zinc was assessed in children with SCD-SS (ages 4 to 10 years) who were randomly assigned to receive 10 mg zinc/d in cherry syrup (zinc) or cherry syrup alone (placebo) for 12 months. Evaluations for growth, dietary intake, and other biochemical parameters were made at baseline and 3, 6, and 12 months. Longitudinal mixed effects analysis evaluated differences between groups over 12 months. RESULTS: A total of 38 prepubertal children (20 male and 18 female; 18 zinc and 20 placebo) completed the study (7.1 +/- 1.7 years old). At baseline, plasma zinc was low (< or = 70 microg/dL) in 7 male subjects. Despite the significant increase in height over 12 months (+0.7 cm) with zinc supplementation (p = .019), plasma zinc did not change over the 12 months of study, and there was no association between plasma zinc and linear growth. Those children with low plasma zinc who received zinc supplementation showed improved linear growth. CONCLUSIONS: These findings suggest that plasma zinc is an insensitive indicator of zinc status in children with SCD-SS. Children with low plasma zinc will benefit from zinc supplementation. However, some children with normal plasma zinc and poor growth may also have growth-limiting zinc deficiency and exhibit a growth response to zinc supplementation. Although this study focused on children with SCD-SS, results may be generalized to other pediatric clinical illnesses where nutrition-related growth failure is investigated. PMID- 16215014 TI - Relationship between feeding difficulties, medical complexity, and gestational age. AB - BACKGROUND: This study examined feeding skills differences in medically complex children born prematurely and full term and sought to determine whether later feeding difficulties within this medically complicated sample were related to medical factors or factors related to preterm status. METHODS: A total of 143 pediatric patients referred to an outpatient clinic for feeding, nutrition, or growth problems were compared by their gestational age at birth across a variety of dependent variables related to feeding development and behaviors. RESULTS: Full-term and preterm children did not demonstrate significant differences in feeding difficulties at first oral feeding. By the time of introduction to solid (cereal) feeding, however, preterm children more often continued to demonstrate difficulty than full-term children. Regression analyses indicated that medical factors, primarily the need for breathing assistance, accounted for more of the variance than prematurity alone. CONCLUSIONS: Medically complex children, especially those with an early need for respiratory support, will benefit from ongoing oral-motor feeding intervention. PMID- 16215015 TI - Standards for specialized nutrition support: adult hospitalized patients. PMID- 16215016 TI - Nutrition in chronic disease management in the elderly. AB - Older Americans experience chronic disease at rates well above other segments of our society. Rates of health services use are also 2 to 3 times that of younger age groups. The most rapidly growing segments of America's aging population are also its most nutritionally vulnerable-women, minorities, and those 85 years of age and older. The routine incorporation of nutrition screening and intervention into chronic disease management protocols will lower healthcare services usage, decrease healthcare costs, help relieve the burden of human suffering experienced by older Americans with chronic disease, and improve quality of life for our nation's elders. PMID- 16215017 TI - Normal aging, nutrition assessment, and clinical practice. PMID- 16215018 TI - Nutrition issues in the home, community, and long-term care setting. PMID- 16215019 TI - Ethical and legal issues in nutrition support of the geriatric patient: the can, should, and must of nutrition support. PMID- 16215020 TI - Medicare and the new medical nutrition therapy benefit. PMID- 16215021 TI - Ins and outs of enteral access: part 2--long term access--esophagostomy and gastrostomy. AB - Access techniques for long-term enteral nutrition (over 4 to 6 weeks) includes cervical esophagostomy and pharyngostomy, gastrostomy, and jejunostomy. Cervical esophagostomy and pharyngostomy are rarely used since the advent of better long term enteral access techniques that are easier to care for and have fewer complications; they are briefly reviewed only for historical completeness and to condemn their use. The different techniques of gastrostomy tube insertion and their indications, contraindications, advantages, and disadvantages are discussed. Part III (to be published in a later issue) of this series will review feeding jejunostomy. PMID- 16215022 TI - The art and science. PMID- 16215023 TI - Enteral feeding: the art and the science. AB - Enteral feeding (TEN) is the preferred method for hospitalized patients requiring nutrition support. However, real and perceived barriers to implementation exist, with gastrointestinal (GI) intolerance being the most common. The purpose of this article is to identify common GI obstacles to effective TEN and to review current practices of TEN delivery and evidence to support such practices. The article reviews the GI anatomy and physiology that affect TEN practices and also identifies related evidence on the most common GI barriers limiting effective TEN delivery. Suggested solutions are provided. PMID- 16215024 TI - Nutrition treatment for HIV wasting: a prescription for food as Medicine. AB - BACKGROUND: The optimal nutrition approach for the promotion of weight gain in HIV-infected adults with wasting remains unclear. Previous dietary interventions report minimal success and provide inadequate information regarding the counseling approach, contact time, session format, and issues addressed with the subject. The methods we report were incorporated in a 12-week intervention trial for the reversal of HIV-wasting. METHODS: All subjects involved in the intervention trial for the reversal of HIV-wasting received weekly, customized, one-on-one counseling and an oral nutrition supplement (480 kcal/d with 30 g protein). The nutrition aims were to (1) increase caloric intake to surpass daily energy requirements by 500 kcal/d (suggested caloric intake: 40 to 50 kcal/kg current weight); (2) increase protein intake (1.6 to 1.8 g/kg current weight per day); and (3) identify foods that may exacerbate or curtail side effects associated with HIV. Also assessed were preconceptions, nutrition knowledge level and primary information source, and obstacles to healthy eating. Sessions, conducted by a nutritionist in an interactive, action-oriented learning approach, ranged from 30 to 60 minutes. RESULTS: At baseline, subjects harbored many misconceptions, reported numerous HIV-related side effects, and lacked practical nutrition strategies, all of which interfered with weight maintenance and health. The protocol strategies were acceptable to the patients (87% subjects completed all visits), with marked improvements in dietary intake, weight, and body composition, both during and after intervention. CONCLUSIONS: We describe a customized nutrition intervention that produces changes in energy intake, maintenance of appropriate protein intake, and the reversal of unintentional weight loss over 5 to 15 months. Sustained improvements occurred across a socioeconomically diverse population, despite persistent disease- and medication associated side effects. PMID- 16215025 TI - Guidelines for the prevention of intravascular catheter-related infections: Centers for Disease Control and Prevention. PMID- 16215026 TI - Branched-chain amino acids in patients with hepatic encephalopathy. 1982. PMID- 16215027 TI - The placement of central venous catheters is a technically challenging procedure with various risks and well-known complications. PMID- 16215028 TI - Surgical treatment for extreme obesity: evolution of a rapidly growing field. PMID- 16215029 TI - Psychological aspects of bariatric surgery. AB - Severe obesity is a serious chronic condition associated with a variety of life threatening medical illnesses. It also has a major impact on mental health and quality of life. A higher than normal prevalence of psychopathology is seen in this population, and there seems to be a direct relationship between body mass index and the degree of psychosocial impairment. This impact on the overall quality of life may have more serious implications on public health than even the clear medical risks of obesity. Among the various psychological disorders seen in the obese treatment-seeking population, there is a higher incidence of depression, anxiety, distorted body image, and eating disorders. This paper reviews the complex psychological issues related to bariatric surgery, offers recommendations regarding presurgical behavioral assessment, and reviews emerging data on the psychological impact of this important new treatment. PMID- 16215030 TI - Altered drug disposition in obesity and after bariatric surgery. AB - The purpose of this review is to evaluate the effect of obesity and gastric bypass on drug disposition. A Medline search was conducted of literature from 1980 onward. All types of literature were included because of the limited data published on this subject. Very few prospective randomized controlled studies are available that have evaluated changes in drug pharmacokinetics and pharmacodynamics in patients who have a body mass index (BMI) >40 kg/m(2) or who have undergone gastric restrictive surgery. Dosing recommendations have been based on general pharmacokinetic principles where specific drug data have not been assessed. Given the increasing prevalence of overweight and obesity in the population, studies that evaluate the impact of obesity on drug disposition, especially related to drugs released in the past 10 years, should be conducted. PMID- 16215031 TI - Metabolic outcomes of gastric bypass. AB - Gastric bypass surgery may be the most successful treatment for morbid obesity. However, postoperative patients are susceptible to deficiencies of several nutrients, including iron, vitamin B(12), and folate. Careful monitoring and supplementation are recommended to prevent negative outcomes of nutrient deficiencies. Metabolic benefits, on the other hand, include improvements in glycemic control and biochemical risk factors for cardiovascular disease. Pursuing surgery as an option for treating morbid obesity requires long-term dedication from the patient and the healthcare team. PMID- 16215032 TI - Roux-en-Y gastric bypass: the dietitian's and patient's perspectives. PMID- 16215033 TI - An update on medical therapy for obesity. AB - Obesity is a risk factor and exacerbates many chronic conditions, particularly diabetes, but also hypertension, hyperlipidemia, heart disease, stroke, certain cancers, arthritis, and obstructive sleep apnea. Despite increased awareness within the public and healthcare system, there has still been a persistent rise in the prevalence of obesity over the past half-century. Failure to halt this epidemic is related to difficulties in adherence to lifestyle changes, forceful counter-regulatory mechanisms opposing weight loss, and the lack of efficient long-term therapy for obesity. This article summarizes the current medical approach to the treatment of obesity, reviewing strategies for lifestyle modification, available pharmacotherapy both as an adjunct to diet and exercise and to ameliorate comorbidities, and an overview of new pharmaceutical agents being developed. PMID- 16215034 TI - Dietary supplements marketed for weight loss. PMID- 16215035 TI - The language and art of policy and politics: neophyte version. PMID- 16215036 TI - Advocacy (public policy) and nutrition support. PMID- 16215037 TI - Ins and outs of enteral access. Part 3: long-term access--jejunostomy. AB - Feeding jejunostomy tubes can be useful in patients with gastroesophageal reflux or a nonfunctioning stomach and can be short-term, long-term, or permanent. These tubes can be inserted surgically, endoscopically, radiologically, or laparoscopically. Each of these techniques is described, and their mortality, morbidity, advantages, and disadvantages are compared. PMID- 16215038 TI - Overview of considerations for the pediatric patient receiving home parenteral and enteral nutrition. AB - Pediatric home nutrition support is a common practice used to provide children with impaired gastrointestinal function and their caregivers a therapeutic environment for the delivery of needed nutrition for proper growth and development. Specialized teams and equipment are now available to make this a successful experience for all involved. This paper will review items specific to the care of infants and children receiving both enteral and parenteral nutrition. PMID- 16215039 TI - C-reactive proteins and chronic disease: what role does nutrition play? AB - The following review provides an update on C-reactive proteins (CRPs) and how they are related to chronic diseases and diet. Emphasis is placed on the mechanism that is involved in the infection/stress-induced formation of CRPs. CRPs' role as biomarkers for coronary heart disease is discussed. This review also discusses the roles that obesity, diabetes, smoking, and synthetic hormones play in increasing serum CRP concentrations. It also summarizes information about how dietary manipulation and exercise can decrease CRP concentrations. The dietary components that seem to lead to a decrease in CRPs include omega-3 fatty acids (fish oil), vitamin E, and moderate amounts of alcohol. Aspirin intake is also discussed as a method that can be used to decrease CRP concentrations. PMID- 16215040 TI - What clinicians can learn from life with Fred. PMID- 16215041 TI - Life with fred: 12 years of home parenteral nutrition. PMID- 16215042 TI - Parenteral and enteral nutrition support groups. PMID- 16215043 TI - Review of a potential role for zinc supplementation in oral rehydration solutions. AB - Patients with short bowel syndrome often present with diarrhea and significant fluid and electrolyte abnormalities. Management consists of the use of medication, dietary modification, and oral rehydration solutions. Zinc deficiency has been shown to result in diarrhea. Oral zinc has been used for the management of acute diarrhea primarily in underprivileged children in developing countries. The effectiveness and feasibility of adding zinc to oral rehydration solutions (ORS) to abate and control diarrhea for adult patients with intestinal failure caused by short bowel syndrome has not been studied. Based on studies evaluating the use of oral zinc supplementation in the treatment of acute diarrhea, recommendations are made for the composition of ORS in adults. PMID- 16215045 TI - Hypercalcemia and peptic ulcer disease-related Milk-alkali syndrome. AB - Milk-alkali syndrome was considered "extinct" by 1985 because of the advent of non-alkaline ulcer medications (ie, histamine-2 receptor blockers and proton pump inhibitors). At that time, it was thought to cause <1% of hypercalcemia, which occurred when one ingested a sufficient quantity of calcium and alkali together. This case emphasizes the importance of considering this syndrome in patients who self-medicate for control of symptoms related to gastroesophageal reflux and peptic ulcer disease and for those using calcium supplementation for prevention or treatment of osteoporosis. PMID- 16215044 TI - The European view of hospital undernutrition. AB - Disease-related undernutrition is significant in European hospitals but is seldom treated or prevented. In 1999, the Council of Europe decided to collect information regarding nutrition programs in hospitals, and for this purpose, a network consisting of national experts from 12 of the Partial Agreement member states was established. The aim was to review the current practices in Europe regarding hospital food provision, to highlight deficiencies, and to issue recommendations to improve the nutritional care and support of hospitalized patients. Five major common problems were identified: 1) lack of clearly defined responsibilities, 2) lack of sufficient education, 3) lack of influence and knowledge of the patients, 4) lack of cooperation between different staff groups, and 5) lack of involvement from the hospital management. To solve the problems highlighted, a combined timely and concerted effort is required from national authorities and hospital staff, including managers, to ensure appropriate nutritional care and support. PMID- 16215046 TI - Enteral nutrition in patients with an open peritoneal cavity. AB - Recent surgical advances have led to the increased survival of critically ill patients requiring postoperative nutritional supplementation. One technique, which has been increasingly used, is that of the open peritoneal cavity. In these cases, the peritoneum is left open, and the viscera are protected with a temporary dressing until the abdomen can be closed. The aim of this study was to evaluate the efficacy and tolerance of enteral nutrition in patients who need open peritoneal cavity management techniques. Patients at a tertiary referral center requiring the use of open peritoneal cavity management who received at least 4 days of enteral nutrition were included in the study. Retrospective data were collected on patients admitted between January 1999 and December 2000, and prospective data were collected on patients between January and May 2001. Energy expenditure and actual caloric and protein intake were determined in all patients. Prealbumin levels and nitrogen balance studies were analyzed when available. Intolerance, defined as diarrhea or gastric reflux, was also evaluated. Average daily total caloric intake was 77 +/- 27%, and average daily protein intake was 68 +/- 24% of estimated needs. Initial serum prealbumin levels were low and remained below normal but increased in some patients during the study. Average nitrogen balance studies from 3 patients was -15 +/- 9.7 g/d. Diarrhea and gastric reflux occurred in 42% and 36% of patients, respectively, and were easily treated. Enteral nutrition can be effectively used in patients requiring open peritoneal cavity management after laparotomy. Overall, enteral nutrition is relatively well tolerated in this patient population. PMID- 16215047 TI - Acid-base disorders and nutrition support treatment. PMID- 16215048 TI - Home enteral tube feeding is becoming an increasing burden on society. PMID- 16215049 TI - Standards of practice for nutrition support physicians. PMID- 16215050 TI - Enteral feeding and bowel necrosis: an uncommon but perplexing problem. PMID- 16215051 TI - Feeding the hypotensive patient: does enteral feeding precipitate or protect against ischemic bowel? AB - Although the enteral route of enteral feeding is the preferred method of nutrition support for critically ill patients, this important therapeutic strategy is not without risk. In human subjects, the digestion and absorption of nutrients induce typical hemodynamic changes, consisting of an increase in mesenteric blood flow at the expense of reduced systemic blood pressure. On rare occasion when providing aggressive enteral nutrition to critically ill patients, common symptoms of gastrointestinal intolerance may progress to a syndrome of abdominal distention, hypotension, and shock, with the development of small bowel ischemia or necrosis. Although the incidence of small bowel ischemia secondary to enteral feeding is low, the overall clinical outcome is still poor and carries a high mortality rate. Enteral feeding is well tolerated and is probably beneficial in most critically ill patients before and after a period of hypotension. Although enteral nutrition may be used with caution during the period of hypotension, evidence of poor gastrointestinal function (increased nasogastric tube output, unexplained abdominal pain, and abdominal distention), or development of dilated loops of bowel or intramural gas (pneumatosis intestinalis) on radiographic studies should be interpreted as potential indicators of gut ischemia. With progress in our understanding of the pathophysiology, diagnosis, and prevention of ischemic injury to the intestinal mucosa, the strategy of aggressive enteral feeding for critically ill patients may result in a reduction in this major complication and enhanced functional recovery from severe illness. PMID- 16215052 TI - Feeding the hemodynamically unstable patient: a critical evaluation of the evidence. PMID- 16215053 TI - Which nutrients are processed by a poorly perfused gut? PMID- 16215054 TI - Kwashiorkor: dysmetabolism versus malnutrition. PMID- 16215055 TI - Aluminum contamination of parenteral nutrition solutions and its impact on the pediatric patient. PMID- 16215056 TI - Total enteral nutrition versus total parenteral nutrition: what do our patients think? PMID- 16215057 TI - Management of the home parenteral nutrition patient. AB - Parenteral nutrition (PN) support can be managed in the home setting for both a short-term and long-term period or for a lifetime, permitting individuals who cannot adequately absorb nutrients enterally to achieve a normal lifestyle. Nutrition support professionals must be aware of home PN (HPN) management principles before discharge to ensure a smooth transition to home with all requisite monitoring. This article will discuss the initiation and monitoring of patients on HPN, the prevention and treatment of potential complications, the contributions of the home infusion provider, and the home nutrition support team and the outcomes of HPN. PMID- 16215058 TI - Update on parenteral iron therapy. AB - The role of parenteral iron therapy has been expanding with the growing use of erythropoietin therapy. Much of the clinical experience regarding the use of IV iron therapy in combination with erythropoietin therapy is based on the hemodialysis patient, but the combination therapy has been used in other patient populations as well. In addition, parenteral iron may be indicated in patients receiving long-term parenteral nutrition and in other clinical situations of iron deficiency when the absorption of iron is impaired or tolerance limited. Once the indication for parenteral iron therapy is established, a selection of the most appropriate agent is required. There are currently 3 parenteral iron preparations available, including iron dextran, sodium ferric gluconate, and iron sucrose. Although all agents have been shown to be effective in correcting iron deficiency, there are differences that exist between them. Both sodium ferric gluconate and iron sucrose have been associated with lower rates of serious adverse reactions than iron dextran, although comparative studies are lacking. In patients with previously documented intolerance to iron dextran, sodium ferric gluconate and iron sucrose have been safely administered. In addition to the immediate and delayed reactions associated with the use of parenteral iron, the risk of iron overload and the potential increased risk of infection are of concern. This article will review the clinical experience of the 3 parenteral iron preparations, discuss safety issues, and provide guidelines on proper dosing and administration. PMID- 16215059 TI - Refeeding syndrome in hospitalized pediatric patients. AB - Refeeding syndrome has been well documented over the years, primarily through case reports and literature reviews. Awareness of refeeding syndrome is crucial in preventing the occurrence of, and the metabolic and physiologic complications associated with, aggressive nutrition support in malnourished populations. Once compromised patients have been identified to be at risk of refeeding syndrome, nutrition rehabilitation should be cautiously initiated. We have found a lack of clinical validation for instituting nutrition support in high-risk pediatric patients who may develop refeeding syndrome. The purposes of our investigation were to determine the incidence of refeeding syndrome in pediatric hospitalized patients beginning on parenteral nutrition and to determine how consistently the Department of Clinical Nutrition standards of care for screening and prevention were followed at our institution. PMID- 16215060 TI - Chest pain associated with total parenteral nutrition infusion. PMID- 16215061 TI - The role of nutrition in the adaptation of the small intestine after massive resection. AB - BACKGROUND: The demonstration that weight gain, growth, and development can be achieved by supplying all essential nutrients exclusively by vein prompted the laboratory evaluation of this parenteral feeding technique in animals that had undergone enterectomy. This study was undertaken to determine the role of nutrition and anabolism in compensation of the small intestine after massive intestinal resection. METHODS: Three or 4 littermate beagle puppies from 10 litters, obtained 8 weeks after weaning and weighing 2 to 3 kg, underwent enterectomy at 10 weeks of age. In the standardized operation, 90% or 95% of the small intestine was removed, as measured from the tail of the pancreas to the ileocecal junction. After enterectomy, the dogs were divided into 2 groups, 1 group being fed exclusively with a kennel diet and the other offered the orally administered diet, whereas all the required nutrients were by vein. After 1 month of continuous infusion, the puppies were weaned from the IV-administered nutrient solution and maintained solely on the standard, orally administered diet. RESULTS: Thirty-four puppies were evaluated for up to 1 year with their control littermates with no resection. Normal growth and development occurred in all puppies after enterectomy during the total IV administration of nutrition. After the parenteral feeding, the dogs with a 90% resection achieved near normal weight gain in contrast to the orally fed dogs with enterectomy that grew to only one half normal size. In the 95% resected group, 5 of 6 dogs survived more than 4 months after parenteral feedings, whereas only 1 of 5 lived more than 1 month while receiving oral feedings alone. At 1 year, the intestine from the parenterally fed dogs that had undergone enterectomy demonstrated increased mucosal cellularity, marked villus hypertrophy, and increased intestinal weight gain per unit length when compared with the orally fed dogs that had undergone resection and the control littermates. CONCLUSIONS: By providing all essential nutrients by vein for 30 days, significant improvement in survival, growth, and nutrition was noted in puppies after massive intestinal resection. PMID- 16215062 TI - A retrospective review of the course of patients with pancreatitis discharged on jejunal feedings. PMID- 16215063 TI - Micronutrients: too much or not enough? PMID- 16215064 TI - Choline deficiency during parenteral nutrition in humans. PMID- 16215065 TI - Vitamin K and nutrition support. PMID- 16215066 TI - Influence of multivitamin regimen on urinary oxalate in home parenteral nutrition patients. AB - BACKGROUND: High urinary oxalate levels have been associated with high ascorbic acid intakes. An alteration in the vitamin regimen for home parenteral nutrition (HPN) patients because of product discontinuation resulted in provision of 500 mg instead of 100 mg ascorbic acid per HPN day. This regimen was associated with high urinary oxalate levels. PURPOSE: To determine if a switch from a multivitamin regimen containing 500 mg to one containing 100 mg of ascorbic acid daily would reduce urinary oxalate levels. METHODS: A 24-hour urine collection for oxalate was analyzed before switching the vitamin regimen back to 100 mg ascorbic acid and repeated 2 months after the change. A paired t test was conducted to compare measurements at baseline and at 2 months. RESULTS: Overall, 18 patients completed both phases of this observational study. The initial urinary oxalate of 517 +/- 63 micromol/day decreased to 425 +/- 47 micromol/day after 2 months (p = .05). However, after applying the exclusion criteria, only 6 patients could be included. The baseline urinary oxalate of 649 +/- 106 micromol/day decreased to 391 +/- 57 micromol/day after 2 months (p = .006). CONCLUSIONS: A change in the parenteral regimen of HPN patients from 500 mg ascorbic acid to 100 mg ascorbic acid is associated with a decrease in urinary oxalate levels. This suggests that a moderate dose of parenteral ascorbic acid (100 mg/day) may limit urinary oxalate appearance in HPN patients. PMID- 16215067 TI - Asymptomatic hypermanganesemia in long-term home parenteral nutrition patients. PMID- 16215068 TI - Hypermanganesemia in an acute care setting. AB - Hypermanganesemia has recently been reported in long-term home parenteral nutrition patients receiving routine trace element supplementation. Because the regulation of manganese involves intestinal absorption and biliary excretion, patients receiving parenteral nutrition, especially those with hepatic dysfunction, may be at risk for manganese toxicity. This case study describes a patient in an acute care setting who had been receiving parenteral nutrition for 7 months. When neurologic symptoms developed, she was found to have a whole blood manganese level almost 3 times normal. Manganese was eliminated from the trace element solution, and a whole blood level was rechecked 3 months later, showing a significant decrease and a marked improvement in facial paralysis and headaches. PMID- 16215069 TI - Clinical applications of L-glutamine: past, present, and future. AB - OBJECTIVE: This review will attempt to summarize recent clinical data on glutamine's use. It will present the concept of glutamine as a "drug" or "nutraceutical," given in addition to standard nutrition support. Key references will be discussed, and clinical recommendations with regard to patients who may benefit and dosing are also provided. RECENT FINDINGS: Glutamine, traditionally considered a nonessential amino acid, now is considered "conditionally essential" after critical illness, stress, and injury. States of illness or injury can lead to a significant decrease in plasma levels of glutamine, and when this decrease is severe, it has been correlated with increased mortality. Laboratory data have demonstrated numerous benefits of glutamine in experimental models of critical illness, cancer, and cardiac injury. The mechanism of these protective effects includes attenuated proinflammatory cytokine expression, improved gut barrier function, enhanced ability to mount a stress response, improved immune cell function, and decreased mortality. Over the last 10 years, clinical trials of glutamine supplementation in critical illness, surgical stress, and cancer have shown benefit with regard to mortality, length of stay, and infectious morbidity. However, data demonstrating a lack of benefit with glutamine supplementation in some patients have been presented as well. It appears that dose and route of administration clearly influence the benefit observed from glutamine administration, with high-dose parenteral glutamine demonstrating an advantage over low-dose enteral glutamine. SUMMARY: High-dose or parenteral (> 0.25 to 0.30 g/kg/day IV or >or=30 g/day enterally) glutamine appears to demonstrate the greatest potential for benefit in hospitalized patients. No evidence of harm has been observed in studies conducted to date; thus, further clinical trials using glutamine as a pharmacologic supplement to standard nutrition are warranted. PMID- 16215070 TI - The data in support of glutamine supplementation. PMID- 16215071 TI - The role of glutamine: counterpoint. PMID- 16215072 TI - Prophylactic pancreatic enzymes to reduce feeding tube occlusions. AB - BACKGROUND: Small-bore, feeding tube occlusion is reported to be as high as 23% to 35%. Pancreatic enzyme solution has been used to dissolve internal clots in feeding tubes. This study examined the prophylactic use of pancreatic enzyme solution with continuous enteral feeding in critically ill patients. METHODS: This was a randomized, unblinded trial conducted in an intensive care unit at a tertiary care hospital. Feeding tubes were randomized to receive pancreatic enzyme solution every 4 hours or usual care. Pancreatic enzyme solution contained lipase 8,000 units, amylase 30,000 units, protease activity 30,000 units, sodium bicarbonate 300 mg, and 5 mL warm sterile water. RESULTS: Ninety-five feeding tubes were followed up in 101 patients. Ten feeding tubes (9.9%) developed primary occlusions. Analysis revealed 2/52 (4%) of the tubes in the enzyme arm had occlusions, whereas 8/49 (16%) of control tubes had occlusions (p = .04). Time to occlusion was significantly longer in the pancreatic enzyme arm (p = .02). CONCLUSIONS: The use of prophylactic pancreatic enzymes may reduce the incidence of feeding tube occlusions. PMID- 16215073 TI - Adverse effects from inappropriate medication administration via a jejunostomy feeding tube. AB - Numerous complications can arise when administering medications to patients receiving continuous enteral feeding. We report a case of a patient who could not be fed by mouth and was receiving continuous jejunal enteral feeding who had an adverse event associated with inappropriate administration of a medication via his jejunostomy tube. He had taken an extended-release niacin product before hospitalization for type IIb hyperlipidemia. The patient was inappropriately given a single dose of 750 mg of niacin as the short-acting tablets that were crushed and administered via the jejunostomy tube. He experienced severe cutaneous flushing, a feeling of warmth, itching, nausea, and emesis. He was noted to have "prickly heat" to the forehead, according to the nursing notes. A discussion of problems and guidelines for medication administration in adult patients receiving continuous tube feeding is provided. PMID- 16215074 TI - Neonatal nutrition: a focus on parenteral nutrition and early enteral nutrition. AB - No population has benefited more from the development and advancement of specialized nutrition support than pediatric patients. Today, neonates comprise the largest group of pediatric patients receiving parenteral nutrition (PN). Nutrient needs of neonates differ substantially from other populations, presenting unique challenges in optimizing nutrition care. Neonates are highly susceptible to catabolic stress because of reduced energy stores and markedly increased energy needs. Immature organ systems and metabolic pathways further complicate the delivery of adequate nutrition in the preterm neonate. Early nutrition support is essential to improve survival, reduce catabolism, promote growth, and limit developmental complications. This article discusses feeding strategies for PN and early enteral nutrition in neonates, particularly the preterm neonate. PMID- 16215075 TI - Maintaining normal blood glucose concentrations with total parenteral nutrition: is it necessary to taper total parenteral nutrition? PMID- 16215076 TI - Effect of acclimation on resting energy expenditure measurements. AB - BACKGROUND: Development of an acclimation protocol for use when measuring resting energy expenditure (REE) would simplify and standardize data collection. The purpose of this study was to determine if our 2 metabolic carts could be used interchangeably and to determine if excluding the first 3 or 5 minutes of data collected as an acclimation period would significantly improve the coefficients of variation (CVs) for oxygen consumed (VO(2)) and carbon dioxide produced (VCO(2)) when performing REE assessments with our metabolic cart systems. METHODS: Thirteen healthy, nonsmoking adults ranging in age from 32 to 45 years, with activity levels ranging from sedentary to highly active, participated. Indirect calorimetry was performed twice in the morning after 30 minutes of supine resting. Subjects had fasted for 12 hours, and did not exercise within the last 24 hours. The system order for testing was randomized for the first measurement. When the first measurement was completed, subjects were crossed over for measurement using a second metabolic cart. RESULTS: The CVs for VO(2) and VCO(2) were significantly lower when excluding the first 3 (VO(2), p = .0005), (VCO(2), p = .0024) or 5 minutes (VO(2), p = .0001, VCO(2), p = .0021) of data compared with no exclusions. No significant differences in CVs between the 3- and 5-minute exclusions were found for VO(2) (p = .3224) or VCO(2) (p = .2255). CONCLUSIONS: Clearly, our machines cannot be used interchangeably within a study. An acclimation period improves CVs of VO(2) and VCO(2.) The similarities in CVs led us to adopt a 3-minute acclimation period for measuring REE. PMID- 16215077 TI - Restoring and maintaining patency of enteral feeding tubes. PMID- 16215079 TI - Indirect calorimetry: applications to practice. PMID- 16215078 TI - Tube feeding in mechanically ventilated critically ill patients: a prospective clinical audit. AB - BACKGROUND: When and whether early enteral nutrition (EN) benefits critically ill patients is debatable. This prospective clinical audit aimed to evaluate the feasibility of an early EN protocol and to identify factors that may hinder EN delivery in critically ill patients. METHODS: Thirty-six medical patients with severe respiratory failure under invasive ventilation and scheduled to receive early EN, with a length of ICU stay >72 hours, were included. As asserted by the Society of Critical Care Medicine, 8% of patients were priority 1, 72% priority 2, and 20% priority 3 for intensive therapeutic and vital support interventions. RESULTS: Overall, because of gastrointestinal complications, only 39% of the prescribed EN was administered; only 8 (22%) patients did tolerate EN within the first 48 hours after admission and did achieve their minimum nutritional requirements. The most frequent complication (78%) was high volume of gastric residuals followed by abdominal distention (61%), both associated with hemodynamic instability (HI). Gastrointestinal dysfunction was associated with high Acute Physiologic and Chronic Health Evaluation II score (p = .01), total calorie intake (p = .02), total carbohydrate intake (p = .02), HI (p = .03), malnutrition (p = .04), volume of IV saline (p = .04), and concurrent vasoactive drug administration (p = .05). CONCLUSIONS: This audit in extremely severe intensive care patients identified several factors that impair gastrointestinal function and preclude EN at any stage, namely early EN. Nutrition management must take into account concurrent therapies, given their potential interference with nutrition and organ function. PMID- 16215080 TI - If the gut works, use it: but what if you can't? PMID- 16215081 TI - Principles of immunology. AB - The immune system, composed of innate and acquired immunity, allows an organism to fight off foreign pathogens. Healthy immunity accomplishes four essential principles: (1) ability to detect and fight off infection; (2) ability to recognize a host's own cells as "self," thereby protecting them from attack; (3) a memory from previous foreign infections; and (4) ability to limit the response after the pathogen has been removed. In an unaltered state, the intricate network of immunologic organs and cells creates an environment for proper host defense. Without adequate execution of immunologic mechanisms, a host is rendered defenseless against pathogens. Conversely, an unchecked immune response can be self-destructive. As a result of either of these untoward sequelae, immune dysfunction can elicit disease states in the host. The goal of this review is to elucidate the characteristics of a healthy immune system, focusing on the principles of immunity and the cells that participate in host protection. We also briefly discuss the clinical ramifications of immune dysfunction. PMID- 16215082 TI - Gut immunology and the differential response to feeding and starvation. AB - Animal and human research has established the beneficial effects of enteral nutrition on the gut. Both the innate and acquired immune systems are bolstered by enteral alimentation. Emerging clinical concepts tie these beneficial effects to a significant influence on the host immune response to stressors during critical illness. In this article, we examine how enteral nutrition impacts gastrointestinal immunity and demonstrate how these changes may affect the body's systemic immune response to nongastrointestinal challenges. This modulatory effect occurs because of changes in the pattern of cytokine secretion and alterations in lymphocyte and neutrophil recruitment. Imbalances in these regulatory mechanisms may be the engine driving hyperresponsiveness to subsequent challenges in the critically ill patient. PMID- 16215083 TI - Maintaining mucosal immunity during parenteral feeding with surrogates to enteral nutrition. AB - Pneumonia and intra-abdominal abscesses are significantly lower in trauma patients receiving enteral feeding compared with those receiving parenteral feeding. Extensive experimental evidence suggests that this is related to maintenance of the mucosal-associated lymphoid tissue, which provides immunologic protection for both the gastrointestinal and respiratory tracts against microbial flora and infectious pathogens. This system is exquisitely sensitive to the route and type of nutrition delivery that affects its functional effectiveness. Although parenteral nutrition decreases the effectiveness of this extraintestinal mucosal immunity, specialty nutrients like glutamine and neuropeptides such as gastrin-releasing peptide and cholecystokinin are capable of preventing some of the immune defects associated with parenteral nutrition. This review examines the mechanisms associated with the mucosal immunity and role of both glutamine and neuropeptides in normalizing defects induced by parenteral feeding. Based upon evolving data, specific nutrients and products of the enteric nervous system show promise as adjuncts to parenteral feeding that are capable of maintaining immune function in patients unable to be fed via the gastrointestinal tract. PMID- 16215084 TI - Is immunonutrition ready for prime time? Two points of view. PMID- 16215085 TI - Probiotics as functional foods. AB - Probiotic products have been widely used in Japan and Europe for years. Probiotics are now emerging as an important category of food supplement in the United States. Questions about the biologic nature, available products, claimed health benefits, and safety and regulation of probiotics are important for both consumers and nutrition professionals. Probiotics can be considered functional foods because they provide health benefits beyond the traditional nutrition function. With few exceptions, most probiotic products currently available contain lactic-acid-producing bacteria, which mainly belong to the genera Lactobacillus and Bifidobacterium. We reviewed the scientific papers published in major nutrition journals and abstracts available on the PubMed website regarding probiotics. Evidence suggests the following beneficial effects of probiotics: normalization of the intestinal microflora, ability to block the invasion of potential pathogens in the gut, prophylactic or therapeutic treatment for several types of diarrhea, relief of symptoms of irritable bowel syndrome and inflammatory bowel disease, amelioration of lactose intolerance, prevention of colon cancer, modulation of immune function, inhibition of Helicobacter pylori, and possible enhancement of calcium absorption and reduction of blood cholesterol levels. Mechanisms for the above benefits have been proposed, but none has been proven. An adequate level of viable bacteria in a probiotic product and an appropriate daily dose are critical to achieve a health benefit. Because probiotics are not known to be pathogenic and their upper tolerable level is high, they could be promoted as a beneficial food supplement. Currently, no disease-prevention or therapeutic claim for probiotics is legally allowed. PMID- 16215086 TI - The role of probiotics in gastrointestinal disease. AB - Patients with intestinal failure often have diarrhea for a variety of reasons. Treatment of diarrhea has centered on the use of antidiarrheal medications and antibiotic therapy. Recently, there has been interest in the oral administration of probiotics for these patients with small bowel bacterial overgrowth, inflammatory bowel disease, and radiation enteritis. This paper reviews the studies that have used probiotics to treat patients with short bowel syndrome and gastrointestinal disorders. Additionally, some of the specific issues regarding probiotics are addressed. PMID- 16215088 TI - Death by parenteral nutrition: fact or fiction. PMID- 16215087 TI - A practical guideline for calculating parenteral nutrition cycles. AB - BACKGROUND: Both physiologic and psychological reasons for cycling total parenteral nutrition (TPN) have been well established. Despite widespread acceptance of this practice, the only previously published method for calculating TPN cycle rates is inherently flawed. METHODS: A mathematical formula was derived to facilitate reliable calculation of cyclic TPN flow rates as a function of total volume and cycle time. A publicly accessible website was subsequently developed to expedite rapid determination of TPN cycles. RESULTS: A fail-safe method of calculating TPN cycle flow rates can be expressed as F = V/(4T-10), where F is equal to the basal flow rate (mL/h), T is equal to the desired cycle time (hours), and V is equal to the total volume of TPN (mL) to be delivered in 24 hours. The basal flow rate and twice the basal flow rate are used for the first and last 2 hours of the TPN cycle, and the remainder of the cycle runs at 4 times the basal flow rate. TPN cycles may be easily calculated online using this formula at http://peds.stanford.edu/tpn.html. CONCLUSIONS: We have developed a fail-safe method of calculating TPN cycle flow rates that will consistently deliver the desired volume and have made an online implementation of this formula publicly available. PMID- 16215089 TI - Bacteremic episodes and copper/zinc ratio in patients receiving home parenteral nutrition. PMID- 16215090 TI - Defining the new gold standard for nutrition support in acute pancreatitis. PMID- 16215091 TI - The pathophysiology of pancreatitis. AB - The pancreas is an exocrine and endocrine gland that is required for normal digestion and metabolism of proteins, carbohydrates, and fats. Acute pancreatitis (AP) is an inflammatory process of the pancreas that involves peripancreatic tissues and remote organs. Mild AP occurs in 80% of patients requiring hospitalization and severe AP occurs in the other 20%. Chronic pancreatitis (CP) is an inflammatory disorder that causes anatomic changes, including infiltration of chronic inflammatory cells and fibrosis of the pancreas. This review discusses biochemical and histologic features of pancreatic injury in AP and CP and some of the important clinical sequelae associated with these abnormalities. PMID- 16215092 TI - Acute pancreatitis: a review of pathophysiology and nutrition management. AB - Acute pancreatitis (AP) is an acute inflammatory process of the pancreas that may involve peripancreatic tissue or remote organs. In its more severe forms, it is a significantly catabolic illness for which specialized nutrition support is required. This review summarizes the pathophysiology of AP, the severity grading systems used in its evaluation, and general treatment strategies, and reviews the value of enteral tube feeding and parenteral nutrition in supporting the patient with this common disease. PMID- 16215093 TI - Severe acute pancreatitis: nutritional management in the ICU. AB - Patients with acute pancreatitis have elevated nutritional needs due to increased energy expenditure and catabolism. It is a clinical challenge to provide adequate nutrition to these patients while maintaining gut function, preventing pancreatic stimulation, and minimizing the risk of septic and metabolic complications associated with nutritional support. We present the case of a patient who had severe acute pancreatitis and was initially given total parenteral nutrition. After a period of initial improvement, he developed hyperglycemia, bacteremia, and sepsis. Parenteral nutrition was discontinued and infection was treated with antibiotics. Subsequent nutritional support consisted of enteral feeding with an elemental diet infused via a nasojejunal feeding tube. His condition improved gradually and he made a full recovery. This case illustrates the difficulties encountered while managing a case of severe acute pancreatitis and provides an evidence based approach to the nutritional management of severe acute pancreatitis in the intensive care unit setting. PMID- 16215094 TI - Enteral nutrition in acute pancreatitis: a survey of practices in canadian intensive care units. AB - INTRODUCTION: Enteral nutrition (EN) is becoming the standard of care for the artificial nutrition support of the patient with severe acute pancreatitis. This study was designed to describe nutrition support practices for acute pancreatitis in Canadian intensive care units (ICUs) and to identify any barriers to the provision of EN in this patient population. METHODS: After an initial letter of invitation, a survey was sent to all Canadian ICUs with a census of > or = 8 beds. At each center, the critical care dietitian was asked 13 questions about usual practice of providing nutrition support to critically ill patients with acute pancreatitis. RESULTS: Out of 62 ICUs successfully contacted through the initial letter that met entry criteria, responses were obtained from 54 (87%). EN was provided to patients with pancreatitis routinely in 13% (7/54) of units, occasionally in 72% (39/54), and never in 15% (8/54) of the ICUs. Technical difficulty obtaining small bowel access, reported by 38 units (72%), and lack of physician support for EN, as noted in 25 units (47%), were identified as the most common barriers to EN in this population. Enteral access was most commonly obtained via the nasojejunal route. The time frame from ICU admission to initiation of EN (when provided) differed widely between centers, varying from up to 24 hours to 48 hours in 22 units (48% of 46 ICUs), 3 to 5 days in 19 units (41%), and >5 days in 5 units (11%). Supplemental parenteral nutrition (PN) was commonly added to EN, routinely at 8 centers (18% of 45 ICUs) and only occasionally in another 20 units (44%). The duration of supplemental PN when used in conjunction with EN was <7 days in 83% (24/29) of the ICUs. When EN was not initiated, PN was used in all but one ICU. CONCLUSION: Although EN is being commonly provided to patients with acute pancreatitis, PN use remains prominent in many ICUs across Canada. Technical difficulty obtaining small bowel access and lack of physician support seem to be the most common barriers impeding use of EN. PMID- 16215095 TI - Management of chronic pancreatitis. AB - Chronic pancreatitis (CP) is an inflammatory disorder that results in permanent impairment of the glandular anatomy of the pancreas with or without functional abnormalities. The pathogenesis of CP is usually unclear, except in the case of alcohol-induced disease. The most common symptoms of CP are abdominal pain, diarrhea, and weight loss often requiring recurring hospitalization. Over time, pancreatic endocrine and exocrine dysfunction may develop as the disease progresses, and a variety of complications can occur. Among the possible complications are nutrient malabsorption and diabetes mellitus. The treatment of CP is difficult and challenging for every physician. Relieving pain is the first step in treating CP. This symptom needs to be controlled, often with narcotics, which can cause dependence. Diarrhea usually indicates the presence of steatorrhea, which is often treated with a high-calorie, high-protein, and low fat diet to minimize symptoms of the underlying disease and to promote weight retention or gain. Pancreatic replacement therapy is used to combat maldigestion and malabsorption. Patients with diabetes may need insulin therapy for glycemic control. The use of parenteral nutrition for bowel rest is a standard approach in patients with symptomatic CP. The use of jejunal enteral feeding recently has been evaluated for efficacy in CP patients. The role of pancreatic endotherapy in the management of CP is evolving. Several reports have suggested that endoscopic therapy aimed at decompressing the obstructed pancreatic duct can be associated with pain relief in some patients. Surgery should be considered in patients who fail medical therapy. PMID- 16215096 TI - Strategies in managing chronic pancreatitis-placement of direct percutaneous endoscopic jejunostomy feeding tubes. AB - Patients with chronic pancreatitis (CP) often have maldigestion and malnutrition. Nutrition support during acute and CP traditionally has been provided by parenteral nutrition. In acute pancreatitis, jejunal feeding may accelerate resolution of the inflammatory process, protect against infection, and improve outcomes at a reduced cost when compared with parenteral nutrition. Jejunal feeding may also be beneficial for patients with CP. Prolonged jejunal access may be achieved via a direct percutaneous endoscopic jejunostomy (DPEJ). This article will review the rationale and evidence for jejunal feeding, indications and contraindictions for DPEJ placement, and the technique and outcomes of DPEJ in patients with CP. PMID- 16215097 TI - Unraveling the mystery of pain in chronic pancreatitis. AB - Over 95% of patients with chronic pancreatitis have intractable abdominal pain. The etiology of the pain remains unclear and is considered multifocal. Treating pain associated with chronic pancreatitis requires a comprehensive approach. This care model encompasses interventional, medical, and adjuvant therapies for both pain and symptom management. Maximizing pain and symptom management in patients with chronic pancreatitis results in timely discharges from the hospital, fewer readmissions, and overall decrease in emergency department visitations. Proper assessment and management of pain and symptoms in patients with chronic pancreatitis improves overall quality of life. PMID- 16215098 TI - Type 1 diabetes mellitus: a window to the present and a view of the future. PMID- 16215099 TI - Management of parenteral nutrition in acutely ill patients with hyperglycemia. AB - Many hospital clinicians manage patients with hyperglycemia receiving nutrition support. Recent studies underscore the importance of tight glucose control in hospitalized patients. Over the short term, hyperglycemia can adversely affect fluid balance, immune function, inflammation, and outcome; glucose control can improve these effects. Because this appears to be true for patients with and without a known diagnosis of diabetes, hyperglycemia should be aggressively treated in all patients. Avoidance or minimization of hypoglycemia also is important. In general, the nutrition assessment, indications for nutrition support, and estimate of nutrition requirements for critically ill patients with hyperglycemia are similar to those of nondiabetic patients. Because overfeeding may cause hyperglycemia, appropriate provision of calories is essential. Although the technology to deliver tight glycemic control in intensive care unit settings is widely available, implementing a safe and effective program requires standardized approaches. Studies testing which insulin infusion safely achieves tight glucose control are limited. Further research should focus on the risks and benefits of parenteral nutrition use in this group of patients, as well as on optimal methods of achieving glucose control. PMID- 16215100 TI - Management of blood glucose and diabetes in the critically ill patient receiving enteral feeding. AB - Diabetes mellitus causes profound alterations in nutrient metabolism, leading to long-term systemic complications, particularly in individuals with poorly controlled disease. Nutrition support clinicians need to be aware of the effect of acute illness on stress-induced and established diabetes mellitus and the appropriate management strategies. Because enteral feeding is a vital component of critical care for many patients with diabetes mellitus, it is important to develop techniques to best manage metabolic control during enteral feeding. This article will review mechanics of blood glucose regulation, evaluation of energy sources for patients with diabetes mellitus, selection of enteral formulas and infusion routes, and glycemic control in patients receiving tube feeding. At this point, it is appropriate to use a standard formula to initiate feedings in most patients with blood glucose abnormalities. Close monitoring and judicious use of insulin are key in maintaining glucose control and avoiding complications. PMID- 16215101 TI - Evidence-based medical nutrition therapy for diabetes. AB - The 2002 American Diabetes Association (ADA) nutrition principles and recommendations are classified according to the level of evidence available using the ADA evidence grading system. Research also supports medical nutrition therapy (MNT) as an effective therapy in reaching treatment goals for glycemia, lipids, and blood pressure. Recommendations for carbohydrate, protein, dietary fat, micronutrients, and alcohol are summarized. The first priority for persons requiring insulin therapy is to identify a food/meal plan that can be used to integrate an insulin regimen into the person's lifestyle. MNT for type 2 diabetes progresses from prevention of obesity or weight gain to improving insulin resistance to contributing to improved metabolic control. The progressive decline in beta-cell failure requires that MNT progress from MNT as monotherapy to MNT in combination with oral glucose-lowering agents to MNT with insulin therapy. Monitoring of outcomes is essential to assess the outcomes of lifestyle interventions or to determine if changes in medication(s) are necessary. PMID- 16215102 TI - Management of diabetic gastroparesis. AB - Diabetic patients with nausea and vomiting need evaluation to determine symptom etiology; diabetic gastroparesis should never be the default diagnosis. If no other etiology for nausea and vomiting is found after an appropriate evaluation, treatment focuses on effectively relieving symptoms while maintaining adequate nutritional status. Therapy for patients with nausea and vomiting consists of restoring volume, glycemic, and electrolyte status and providing antiemetics generously in both a remittive and prophylactic fashion. Prokinetics are best used to treat gastric stasis or clinically significant delayed gastric emptying. A gastrostomy is rarely indicated, but a jejunostomy may be helpful in maintaining nutrition. Parenteral nutrition should generally be avoided because of high complication rates in this setting. Surgical intervention is essentially unstudied in this population and cannot be recommended. Gastric electrical stimulation is an interesting and novel therapy, but its efficacy beyond placebo remains to be conclusively shown. PMID- 16215103 TI - Pancreatic islet cell transplantation: a treatment strategy for type I diabetes mellitus. AB - Type I diabetes mellitus continues to be a serious disease with long-term morbidity from complications of the disease, despite better knowledge of physiology and better patient care. Pancreatic islet cell transplantation, still experimental, is gaining acceptance as a treatment strategy for individuals with type 1 diabetes who have poor glucose control and frequent hypoglycemic episodes despite maximal care. The procedure can improve glucose control, avoiding further hypoglycemic episodes, and can achieve insulin-independence at the expense of immunosuppression treatment. Islet cell transplantation does not correct all the metabolic abnormalities seen in type 1 diabetes, and glucose tolerance can remain abnormal. The dietary management of the patients still needs to be defined. Further research is focused at improvement in islet isolation and the refinement of immunosuppression strategies aiming toward immune tolerance, bringing hope for a cure of diabetes. PMID- 16215104 TI - Posttransplant diabetes mellitus: cause, impact, and treatment options. AB - The incidence of diabetes mellitus (DM) is increased in adult organ transplant recipients. As many as 30% to 45% of solid organ adult transplant patients have DM before transplantation or develop posttransplant diabetes mellitus (PTDM). Risk factors for PTDM include family history, ethnic or genetic background, insulin resistance, and diabetogenic effects of immunosuppressive medications. Posttransplant hyperglycemia may result in increased platelet aggregation, increased wound infections, dehydration, and loss of lean body mass. More significantly, long-term complications of DM such as coronary artery disease and peripheral vascular disease may be exacerbated with the use of immunosuppressive medications whose known side effects include hyperglycemia, hyperlipidemia, and hypertension; these effects may lead to premature transplant graft dysfunction. Treatment goals for PTDM reflect those of the American Diabetes Association guidelines; long-term management is linked with early, patient-centered education and optimizing minimally diabetogenic immunosuppressive medication regimens. A multidisciplinary team including the patient, family/support people, transplant surgeon, transplant physician, transplant nurse coordinator, transplant social worker, pharmacist, dietitian, and diabetes educator is crucial to long-term management of PTDM. PMID- 16215105 TI - New drugs for management of diabetes: insulin analogues, inhaled insulin, pramlintide, and novel peptides. AB - Diabetes mellitus has reached epidemic proportions and contributes to considerable morbidity and mortality in the form of metabolic, microvascular, and macrovascular complications. Although there is no cure, large controlled studies demonstrate the importance of strict glycemic control in reducing progression of microvascular disease and associated morbidity. Insulin replacement is necessary for all patients with type 1 diabetes. In treatment of type 2 diabetes, more timely addition of insulin as patients fail to attain glycemic targets on combinations of oral agents has become widely accepted. Pharmacokinetic properties of human insulins limit their ability to mimic physiologic insulin secretion. Analog insulins (prandial and basal) are designed with improved physiologic pharmacokinetic characteristics to enable more simplified insulin dosage adjustments and a reduced risk for hypoglycemia. Inhaled administration has emerged as a promising route for noninvasive prandial insulin. Clinical trials are under way to further characterize safety and efficacy of inhaled insulin preparations. Despite numerous pharmacologic advancements, the majority of diabetic patients continue to have inadequate glycemic control. New information regarding biochemistry and pathophysiology of the disease is providing exciting opportunities for drug development. Promising new therapeutic classes include the synthetic analog of amylin (pramlintide), glucagon-like peptide (GLP-1) derivatives, and dipeptidyl peptidase IV inhibitors. Intriguingly, GLP-1 hormones may have important biologic actions aside from stimulating insulin release, including inhibition of gastric motility and acid secretion, suppression of glucagon secretion, and islet cell proliferation. Although additional studies are needed, perhaps these emerging agents will have greater efficacy and safety because of a higher degree of selectivity for their molecular targets. PMID- 16215106 TI - Utility of intensive blood glucose control: generalizable to all general surgery patients? PMID- 16215107 TI - Nutrition support in mechanically ventilated, critically ill adult patients: are we ready for evidence-based clinical practice guidelines? PMID- 16215108 TI - Arginine: mediator or modulator of sepsis? AB - Arginine is a conditionally essential amino acid that plays pivotal roles in maintaining body homeostasis. Arginine is a substrate for protein synthesis but can also be metabolized to various bioactive compounds that include nitric oxide, ornithine, polyamines, creatine phosphate, agmatine, and dimethylarginines. Arginine produces physiologic effects via nitric oxide dependent and independent pathways. Nitric oxide is important for the modulation of vascular tone, inflammation, immune function, endothelial function, platelet and leukocyte adherence, and neurotransmission. Nitric oxide modulates many biochemical processes important for the response to sepsis. Arginine, independent of nitric oxide, is important for growth, wound healing, cardiovascular function, immune function, inflammatory responses, energy metabolism, urea cycle function, and other metabolic processes. Arginine supplementation improves outcomes in animals with sepsis, wounds, ischemia-reperfusion injury, and following thermal injury. Enteral administration of arginine improves endothelial function but has little effect upon hemodynamics during human sepsis. An analysis of clinical studies using enteral formulas with supplemental arginine suggests benefits upon outcome, with no evidence of significant detrimental effects. PMID- 16215109 TI - A rational use of immune enhancing diets: when should we use dietary arginine supplementation? AB - Controversies in any arena of human activity often result in the polarization of the individuals involved into 2 opposing camps. Controversy about the use of immune-enhancing diets (IEDs) is no exception. On one hand, some groups are proposing indiscriminate use of IEDs, whereas others have created guidelines advocating that their use should be banned for the critically ill. At stake is an emerging paradigm: that dietary manipulation of the immune system is possible and may become an important adjunct to other therapies, thus helping prevent or treat multiple diseases for millions worldwide. Under these circumstances, extremist claims of miraculous benefits or inappropriate assertions of evil can only delay the emergence of a nascent science. This paper is therefore a plea for moderation from both camps, lest we cause irreparable damage to our clinical practices and potential injury to individual patients. IEDs all contain arginine. However, they also contain other substances such as omega-3 fatty acids, and nucleotides. The use of all these nutrients together into commercial IEDs without adequate evaluation of their individual effects has prevented the development of mechanistic hypotheses of action. Despite this, IEDs have been tested extensively, allowing the development of guidelines for their use. IEDs should be used for surgical patients, especially those undergoing elective surgery. IEDs show no benefit and indeed can potentially harm patients with sepsis, especially in the nonsurgical group, and should not be used outside of research protocols. Advances in basic research have helped us understand mechanisms of how arginine contained in IEDs may help surgical patients but may be deleterious in patients with sepsis. A review of the basic mechanisms of action of arginine on the immune system is enclosed in this paper and should serve as a basis for the development of scientific principles that guide clinical use of IEDs. PMID- 16215110 TI - Enhancing the response to parenteral nutrition in critical care. AB - Parenteral nutrition (PN) is an essential component in the support of critically ill patients with gastrointestinal dysfunction. Although PN cannot fully reverse hypermetabolism and accelerated skeletal muscle breakdown observed during periods of critical illness, it can prevent the adverse effects associated with malnutrition. The use of PN is not without complications, so care must be taken to ensure successful clinical outcomes with this complex therapy. Strategies have been developed by practitioners to promote safe practices with implementation of PN therapy and optimize a patient's response to PN. Identification of appropriate patient populations, strict glucose control, and manipulation of macro- and micronutrients are techniques being used to augment a patient's response to PN administration. This article will review the novel methods used to enhance benefits received from PN during critical illness. PMID- 16215111 TI - Hyperglycemia and nutrition support: theory and practice. AB - Hyperglycemia has long been known to portend a worse short-term prognosis than euglycemia in hospitalized patients. Hyperglycemia reflects illness severity and results in deleterious consequences for certain patient groups. In a surgical intensive care unit population, normalization of morning glucose values using insulin infusions improved mortality vs controls. Until additional studies in other patient groups are reported, practitioners of nutrition support must prevent excessive iatrogenic hyperglycemia, in particular when using parenteral feeding, and should choose aggressive glycemic goals that also avoid hypoglycemia. PMID- 16215112 TI - Specialized nutrition support in the hospitalized obese patient. AB - The rising prevalence of obesity in the United States has resulted in concerns regarding the implementation of safe and effective specialized nutrition support in hospitalized obese patients. This review discusses complications associated with the provision of specialized nutrition support for obese patients and emphasizes the scientific basis for and against the provision of hypocaloric, high-protein parenteral and enteral nutrition support. Because there is a paucity of data concerning enteral nutrition support, some practical aspects of implementing hypocaloric high-protein enteral nutrition are also given. PMID- 16215113 TI - Indications for administration of parenteral nutrition in adults. AB - BACKGROUND: Who benefits from parenteral nutrition (PN) has been the subject of much debate and 4 recent meta-analyses. METHODS: We reviewed the 4 meta-analyses that examined the prospective, randomized, clinical trials (PRCT) that compared PN with no nutrition support (standard care) for design, study population, outcomes evaluated, and results. RESULTS: Overall, a total of 113 PRCT were included in the 4 meta-analyses; however, only 4 studies were included in all of them. Despite the differences in populations studied and outcomes evaluated, some similarities emerged: (1) PN does not affect mortality; (2) PN does not reduce complications in normally nourished patients; (3) in malnourished patients, PN demonstrated a trend for reduced infections and complication rates; and (4) PN reduced postoperative complications in patients having surgery for cancer of the esophagus or stomach. CONCLUSION: PN does not appear to be beneficial for most hospitalized patients. Among those with malnutrition or with upper gastrointestinal cancer, benefits may exist; however, these were influenced by quality of the study and year of publication. PMID- 16215114 TI - Anabolic agents: adjuncts to nutrition support. AB - Anabolic agents as adjuncts to nutrition support therapy have been used to improve malnutrition and establish anabolism. Growth hormone, insulin-like growth factor, and anabolic steroids have been studied for their potential to reverse the catabolic process and promote anabolism. This paper reviews several anabolic agents and their possible role in nutrition support therapy. PMID- 16215115 TI - Parenteral nutrition-associated hepatotoxicity. AB - BACKGROUND: Parenteral nutrition has been associated with the development of several hepatobiliary complications. The aim of this article was to review the current available literature associated with parenteral nutrition-induced hepatotoxicity. METHODS: We performed a literature search and reviewed all related articles published within the past 5 years, from 1999 to 2003. RESULTS: As a result of this review, we were able to determine the suspected etiology and pathogenesis, commonly used diagnostic tests and procedures, and current methods used for prevention and treatment of parenteral nutrition-associated hepatotoxicity. CONCLUSIONS: Regardless of current advances in research, much is left to be determined in the areas of etiology, pathogenesis, prevention, and treatment of parenteral nutrition-induced hepatobiliary complications. Practitioners should be encouraged to participate in further research, particularly in studies involving human subjects. PMID- 16215116 TI - Predicting energy needs in obese patients. AB - Obesity has become an epidemic in the United States, with other western countries also reporting increases in incidence of obesity. With many associated comorbidities, it is the most common nutritional disorder facing the medical team. However, the assessment of macronutrient needs for nutrition support regimens in obese adults is controversial. This review summarizes existing research on popular predictive approaches, including the Harris-Benedict equation, kilocalories per kilogram, and the Ireton-Jones equations. Complications including special considerations for patients who have undergone bariatric surgeries and current evidence on hypocaloric regimens are also discussed. PMID- 16215117 TI - Nutrition support in the morbidly obese, critically ill patient. AB - Nutrition support in the critically ill patient is challenging but is even more difficult in a morbidly obese patient. This case report chronicles the care of a 6-foot-tall, 256-kg male (body mass index 76.5 kg/m(2)) who spent over a month in the intensive care unit for respiratory failure, sepsis, and acute renal failure. Parenteral nutrition was provided throughout his critical care course. One of the major difficulties encountered was determining his nutritional needs. A hypocaloric nutritional regimen was used, along with moderate protein provisions. Numerous electrolyte imbalances occurred, including hypercalcemia that did not resolve by eliminating calcium from the parenteral nutrition solution. Enteral nutrition was desired but was not used initially because of a need for vasopressors, a diagnosis of pancreatitis, difficulty in documenting feeding tube placement because of diagnostic limitations secondary to the patient's large size, and concern about managing stools. Eventually, oral intake and supplemental enteral feeding were initiated. Nutrition support team members worked closely with the interdisciplinary care team to develop strategies to manage the nutritional problems related to his obesity. A discussion of the various nutritional issues encountered in the care of this patient is provided. Reasonable nutritional status was achieved, but this case reflects some of the challenges encountered in caring for the nutritional needs of select patient populations in clinical practice and the need for increased research and guidelines in this area. PMID- 16215118 TI - The lived experience of home total parenteral nutrition: an online qualitative inquiry with adults, children, and mothers. AB - BACKGROUND: The purpose of this study was to access a sample of short-bowel patients receiving home total parenteral nutrition (HTPN) in their home environment and describe their HTPN experience using qualitative methodology. METHODS: Three semistructured, open-ended, in-depth interviews were conducted online over a 5-month period with 6 adults (aged 27 to 45 years) who received HTPN for 2 to 8 years, 3 children (aged 8 to 10 years) receiving HTPN since birth, and the children's mothers. The constant comparison method was used to code and categorize data, inductively develop themes, and create a taxonomy (flow chart) of themes from the data. Credibility of data analysis and themes was established by the prolonged engagement with subjects and subject review of themes identified. RESULTS: Six major themes emerged: affirmation of life, infusion-related complications, lifestyle adaptations, self-worth, isolation, and food intake. Within most themes, subjects described similar concerns and reactions. A clear difference occurred within the theme of food intake. Adults reported feeling constantly hungry and challenged with resisting temptation to eat restricted foods. They admitted to lack of consistency in following dietary restrictions and repeated attempts to consume contraindicated foods. Children expressed lack of desire for food and mothers complained of their children's refusal to eat. CONCLUSIONS: Using qualitative methods allowed insight into the HTPN experience and depiction of the daily challenges of being total parenteral nutrition-dependent from patients' and caregivers' perspectives. It provides insight into their reactions and behaviors. By gaining this type of awareness and understanding, clinical protocols and services can be planned that are more likely to derive successful outcomes. PMID- 16215119 TI - Copper deficiency during parenteral nutrition: a report of four pediatric cases. AB - The standard of care for patients with cholestasis (direct bilirubin >or=2 mg/dL) while receiving parenteral nutrition (PN) solutions is to reduce or discontinue the copper and manganese. The repercussions of this action have not been studied. Two adult case reports document low serum copper levels associated with clinical symptoms of copper deficiency after the removal of copper from their PN solutions. We now describe the first known series of pediatric patients to develop copper deficiency after copper was removed from their PN solutions. PMID- 16215120 TI - Clinical controversies: the role of glutamine counterpoint. PMID- 16215121 TI - Kwashiorkor: dysmetabolism versus malnutrition. PMID- 16215122 TI - Accessing the keys to life-sustaining treatment. PMID- 16215123 TI - The biology of human immunodeficiency virus infection. AB - The aim of this article is to review the basic biology of infection with HIV-1 and the development of the acquired immunodeficiency syndrome. The discussion will include epidemiology, general description of the retroviruses, pathogenesis of the immune deficiency, clinical consequences, treatment, and treatment outcomes. Aspects of the infection that affect protein and energy balance will be identified. PMID- 16215124 TI - Evaluation of nutrition assessment parameters in the presence of human immunodeficiency virus infection. AB - Malnutrition, weight loss, and body cell mass (BCM) depletion have long been observed in individuals with HIV infection and acquired immunodeficiency syndrome and are associated with increased morbidity and mortality. Nutritional status may be adversely affected by many factors, including inadequate intake, gastrointestinal disturbances, altered nutrient metabolism, and medication related side effects. Increasing numbers of HIV-infected individuals are exhibiting fat redistribution characterized by any combination of changes including peripheral fat atrophy, visceral or dorsocervical adiposity, or other changes, and these changes have been shown to be associated with increased cardiovascular disease risk. A complete nutrition assessment is key to effective patient management in HIV infection. This review focuses on the evaluation of anthropometry (including BCM and fat redistribution), biochemistries, medication related issues, medical/social/dietary histories, and physical examination and clinical findings. A discussion of BCM, its significance as a parameter of nutritional status, and methodological issues concerning its measurement by available technologies is presented in light of current research. Methods for assessing fat redistribution are discussed. Recommendations for biochemical assessment and medical, social, and dietary histories are reviewed. Finally, medication-related side effects for common antiretroviral therapies and clinical findings that may affect nutritional status are presented. Clearly, nutrition assessment in HIV infection is a complex process involving many factors. PMID- 16215125 TI - A review of nutrition in human immunodeficiency virus infection in the era of highly active antiretroviral therapy. AB - BACKGROUND: Nutrition complications of HIV infection, including wasting syndrome, nutrient deficiencies, and metabolic complications, have been well documented over the last 25 years. METHODS: A systematic review of the literature was performed using the keywords HIV; nutrition; nutrition support; vitamins A, B, C, and E; selenium; zinc; and glutamine through MEDLINE using Medscape and PubMed. RESULTS: Although no accurate prediction equations exist for determining energy needs in patients with HIV/acquired immunodeficiency syndrome (AIDS), the Harris Benedict equation with a 1.3 stress factor has been used for weight maintenance. Some experts recommend protein requirements of 1.0 to 1.4 g/kg for maintenance and 1.5 to 2.0 g/kg for anabolism. There is a general consensus that all individuals with HIV benefit from a daily multivitamin and mineral supplement at levels of 100% of the US recommended daily intake (RDI). Nutrition therapy for HIV wasting is similar to that for other chronic diseases and begins with nutrition counseling. For persistent weight loss, standard oral supplements may be useful. The benefit of specialized supplements has yet to be proven. Enteral and parenteral nutrition support has produced positive outcomes related to mortality and quality of life in patients with HIV. CONCLUSIONS: Although the benefit of providing adequate amounts of calories, protein, and micronutrients for persons with HIV is well accepted, the exact amounts of nutrients and optimal feeding modalities are less clear. Long-term clinical trials are needed to provide more conclusive data on nutrition intervention in HIV infection, particularly related to supplementation of specific nutrients. PMID- 16215126 TI - Gastrointestinal manifestations of AIDS. AB - Although gastrointestinal complications have decreased dramatically in patients with acquired immunodeficiency syndrome (AIDS) who received highly active antiretroviral therapy, these disorders still occur in those who do not seek healthcare, those with resistant virus, and those in developing countries where these drugs are not available. Opportunistic infections are the most frequent causes of gastrointestinal disease in patients with AIDS. A practical approach based on the character of the complaints and location of symptoms will help direct the most appropriate evaluation. In most patients, a diagnosis should be sought because effective antimicrobial therapy is available for most infections in these patients. When possible, improvement in immune function forms an integral part of the treatment regimen. PMID- 16215127 TI - Ethical issues in nutrition and human immunodeficiency virus. AB - Ethical dilemmas often arise in the provision of healthcare. In view of the advances in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) medical management since the late 1990s, many of the challenges and ethical dilemmas encountered by healthcare professionals have changed from terminal or end-of-life issues to long-term management issues. This article presents 5 case scenarios on common ethical issues faced by those providing nutrition care to people living with HIV/AIDS and addresses key questions to ask in the ethical deliberative process. Ethical dilemmas surrounding the use of complementary and alternative medicine, providing aggressive nutrition support, dealing with third-party payers for supplement reimbursement, and managing patients with mental health disorders and substance abuse issues are discussed, with possible solutions presented from an ethics point of view. PMID- 16215128 TI - Treatment of HAART-induced lactic acidosis with B vitamin supplements. AB - Lactic acidosis, a rare but life-threatening condition is fairly common in HIV infected individuals. The cause of lactic acidosis appears to stem from the use of HAART, causing mitochondrial dysfunction and the depletion of flavoprotein cofactors necessary for electron transport. Deficiencies of riboflavin or thiamin can contribute to the development of hyperlactic acidemia. Further, the high incidence of liver disease (hepatitis B or C and alcoholic liver disease) in this population predisposes HIV patients to metabolic abnormalities. Supplementation with thiamin or riboflavin, depending on the individual patient's condition, can reduce elevated lactic acid levels. PMID- 16215129 TI - Practical approach to metabolic assessments in the pediatric human immunodeficiency virus outpatient clinic. AB - BACKGROUND: Metabolic assessment is not a routine part of the care of pediatric HIV-positive patients in the outpatient setting, although research studies are demonstrating an increasing prevalence of metabolic abnormalities in this population. Addition of metabolic assessments to routine clinic care can be hindered by the perception that collection of the data needed for the assessments will need too much clinic time and too many clinic resources to be practical. METHODS: A method to provide metabolic assessments using minimal clinic resources while offering educational opportunities for university students was developed and implemented at the University of Florida Pediatric Immunology and Infectious Diseases weekly clinic. The success rate of metabolic assessment data collection from all clinic patients and the impact of data collection on the flow of clinic activity were monitored for a 12-month period. RESULTS: The metabolic assessment team interviewed 87% of all patients who came to the 454 clinic during the 12 month period and performed a complete metabolic assessment for 75% of the HIV positive patients interviewed. Addition of metabolic assessments to the routine care of each patient did not result in any noticeable adverse effect on the functionality of the clinic. CONCLUSIONS: Addition of metabolic assessments to the routine clinic care of pediatric HIV positive patients can be an educational opportunity for university students and provide the clinic staff with changes in parameters indicative of metabolic abnormalities and without noticeable changes in maintaining the patient schedule at each clinic. PMID- 16215130 TI - The treatment of human immunodeficiency virus-associated wasting syndrome with recombinant human growth hormone, then and now: a pivotal paper review. PMID- 16215131 TI - Combination megestrol acetate, oxandrolone, and dietary advice restores weight in human immunodeficiency virus. AB - BACKGROUND: We examined changes in total body weight (TBW) and health-related quality of life (HRQL) during prolonged combination weight-gaining therapy and dietary advice in HIV. DESIGN: This was a cohort study of patients initially randomized to single agent therapy for 2 months, megestrol acetate (800 mg daily), or oxandrolone (10 mg twice daily), followed by both agents and dietary advice for 5 months. METHODS: Two community health clinics and 1 urban infectious disease clinic were included, as were HIV-positive adult patients receiving highly active antiretroviral therapy with documented 5% weight loss. TBW and HRQL were measured after 7 months (7 m). RESULTS: Twenty-nine of 39 participants completed 7 m. The average sample age was 40 years, 75% were male, and 52% were of color at enrollment. Baseline mean TBW and body mass index (BMI) were 62.5 kg and 21 kg/m(2), respectively. Net gains in TBW, lean body mass, and fat during the 7 m were 5.3 kg (8.5% of baseline), 2.1 kg, and 3.1 kg, respectively (p < .01 for each). BMI increased to 23.1 kg/m(2) (p < .01). Dietary intake increased by 467 kcal/day (p = .03). Physical health improved by 5.7 (100-point scale, p < .01), and mental health was unchanged (-4.2, p = .11). In multivariable models, female gender (p < .01), lower baseline HIV viral load (p = .03), and increasing age (p < .01) were associated with TBW gain. Injection drug use (p < .01) and higher baseline HIV viral load (p < .01) were associated with reduction in physical health. CONCLUSIONS: Prolonged combination therapy with megestrol acetate, oxandrolone, and dietary advice could reverse weight loss and low BMI associated with incomplete viral suppression and improve physical health. PMID- 16215132 TI - Pediatric assessment scale for severe feeding problems: validity and reliability of a new scale for tube-fed children. AB - BACKGROUND: This study reports data on the validity and reliability of a new parent report measure, the Pediatric Assessment Scale for Severe Feeding Problems, designed to assess progress in the development of oral eating skills for children who need prolonged tube feeding. METHODS: The questionnaire was completed by parents of 3 groups of children. The first group consisted of 17 children who received all of their nutrition by tube feedings, a second group of 30 children who were oral eaters but required supplementation by tube feedings, and a third group of 27 children who were referred for feeding difficulties but were not receiving any tube feeding. A subset of parents from each group completed the measure a second time approximately 2 to 4 weeks after completing the initial questionnaire in order to examine the reliability of the measure. RESULTS: The mean scores (scale of 0 to 66) on the measure were 7.3 for completely tube-fed children, 30.0 for partially tube-fed children, and 46.8 for referred, non-tube-fed children. The correlation between first and second administration of the measure was .98. CONCLUSIONS: The validity and reliability of the Pediatric Assessment Scale for Severe Feeding Problems appears to be adequate. The new measure should allow clinicians to better rate initial severity of feeding difficulties and to track the progress of children as they advance from being totally tube-fed to completely orally fed. PMID- 16215133 TI - Massive gastric dilation and necrosis in anorexia nervosa: cause or effect? AB - A 26-year-old female with a diagnosis of anorexia nervosa was admitted and found to have massive gastric dilation and gastric necrosis. Imaging studies suggested the possibility of superior mesenteric artery (SMA) syndrome. She was successfully managed with prompt gastric decompression and was able to resume oral nutrition. Gastric dilation and necrosis may be seen in anorexia nervosa as either an independent event or an SMA syndrome. The SMA syndrome may also be present as either an incidental finding or a true pathophysiologic entity. Finally, significant foregut dysfunction may be mistaken for an eating disorder. Although there is clearly an association between gastric dilation, the SMA syndrome, and eating disorders, cause and consequence may not always be straightforward. Prompt recognition and conservative management are advocated in the absence of abdominal catastrophe. PMID- 16215134 TI - A.s.p.e.N. Statement on aluminum in parenteral nutrition solutions. PMID- 16215135 TI - Monitors in nutrition support. PMID- 16215136 TI - Monitoring therapeutic interventions in critically ill septic patients. AB - Sepsis is the leading cause of admission to intensive care units in the United States. Although the treatment of sepsis is complex and multimodal, nutrition support plays an important role in the management of these patients. The diagnosis of sepsis, disease category, and severity of illness and the change in sepsis severity and organ function over time affect the delivery of nutrition support. This paper reviews the diagnostic criteria of sepsis, the use of "sepsis biomarkers," and regional and global markers of organ function in sepsis and quantitative measures of illness severity and organ dysfunction. PMID- 16215137 TI - The validity of bioelectrical impedance models in clinical populations. AB - Bioelectrical impedance analysis (BIA) is the most commonly used body composition technique in published studies. Herein we review the theory and assumptions underlying the various BIA and bioelectrical impedance spectroscopy (BIS) models, because these assumptions may be invalidated in clinical populations. Single frequency serial BIA and discrete multifrequency BIA may be of limited validity in populations other than healthy, young, euvolemic adults. Both models inaccurately predict total body water (TBW) and extracellular water (ECW) in populations with changes in trunk geometry or fluid compartmentalization, especially at the level of the individual. Single-frequency parallel BIA may predict body composition with greater accuracy than the serial model. Hand-to hand and leg-to-leg BIA models do not accurately predict percent fat mass. BIS may predict ECW, but not TBW, more accurately than single-frequency BIA. Segmental BIS appears to be sensitive to fluid accumulation in the trunk. In general, bioelectrical impedance technology may be acceptable for determining body composition of groups and for monitoring changes in body composition within individuals over time. Use of the technology to make single measurements in individual patients, however, is not recommended. This has implications in clinical settings, in which measurement of individual patients is important. PMID- 16215138 TI - Monitoring energy metabolism with indirect calorimetry: instruments, interpretation, and clinical application. AB - Indirect calorimetry is the best measure to guide calorie administration during nutrition support. This article presents an update of the types of currently available indirect calorimeters and reviews the recent advances that guide the clinical application of indirect calorimetry. The emphasis of this report is placed on issues that the practicing clinician can use to evaluate, interpret, and apply measurements of energy expenditure. PMID- 16215139 TI - Enhancing interpretation of gastric residual volume by refractometry. AB - The traditional practice of gastric residual volumes (GRVs) is flawed in its design and conception, is poorly standardized in its technique, is an inaccurate measure of gastric emptying, and serves as an insensitive marker for regurgitation and aspiration. The refractive index of a solution (like an enteral formula) is a physical property of that solution, which is remarkably constant and reproducible under varying conditions of concentration, pH, and temperature. Refractometry may be performed quickly and easily at the bedside, requires only a small representative sample of aspirated solution, and provides valuable measurements that can be used to calculate both the true total volume of contents and the specific volume of formula remaining in the stomach. Refractometry complements the use of GRVs as a monitor for patients receiving enteral feeding and should improve the accuracy with which patients at risk for aspiration may be identified. PMID- 16215140 TI - The Mini Nutritional Assessment as an assessment tool in elders in long-term care. AB - The prevalence of malnutrition increases with age and is most common in the institutionalized individual. Malnutrition is a condition associated with greater susceptibility to infection, longer hospital stay, and increased mortality. Detection of risk of malnutrition in elders and early intervention may lessen these negative consequences. A tool that has been used for assessing nutritional status of elders is the Mini Nutritional Assessment, or MNA. The MNA tool was originally validated in relatively healthy elders in France and New Mexico, but assessment scores seem to correlate with immune function, morbidity, and mortality of elders in long-term care. One of the advantages of using the MNA for assessing nutritional status of elders in long-term care is that it does not need any biochemical tests, such as serum albumin or prealbumin. A weakness of the MNA is that a number of questions target independent-living elders but not elders in long-term care or elders receiving nutrition support. The benefits, limitations, and interpretations in the use of this tool in a long-term care setting are discussed. PMID- 16215141 TI - Percentage of weight loss as a predictor of surgical risk: from the time of Hiram Studley to today. AB - Hiram Studley's 1936 article of research was the first publication to present a connection between preoperative weight loss and adverse postoperative outcome. Almost 70 years later, weight loss remains one of the most prominently used tools to assess nutritional status and predict surgical risk. This paper provides an overview of surgical practices at the time of Dr Studley and demonstrates Studley's unique contributions to the field of nutrition support. The search for more accurate methods of preoperative nutrition assessment is traced to show how subsequent research continues to validate the use of weight loss in the assessment of surgical risk. New developments center on techniques of body composition assessment to quantify weight lost as functional weight and clarify the impact of malnutrition on operative outcome. PMID- 16215142 TI - Permissive underfeeding of the critically ill patient. AB - The rise in the popularity of nutrition support in the 1970s was associated with the concept of "hyperalimentation." This concept was based on the early findings that increased metabolic rates were observed in various disease states such as trauma, sepsis, and burns. The aim was to feed 40% to 100% above the basal metabolic rate to avoid weight loss associated with critical illness. Since that time, several observations have indicated that permissive underfeeding may be beneficial because: (a) the metabolic rate is not markedly increased in most patients with critical illness except burns; (b) weight gain during nutrition support in critical illness is not caused by a gain in nitrogen but fat; (c) energy intake as glucose in excess of needs causes increased carbon dioxide production and a fatty liver; (d) hyperglycemia increases the risk of infective complications; and (e) a controlled trial of preoperative nutrition in which patients received 1000 kcal above the metabolic rate increased infectious complications. PMID- 16215143 TI - Defining and assessing tolerance in enteral nutrition. AB - Nutrition support has become widely recognized as an essential component of optimal care for acutely ill patients. Enteral nutrition is preferred over parenteral routes when possible. However, prescribed enteral nutritional regimens are sometimes met with side effects and even complications. These adverse events have been collectively termed "intolerance," and forms of intolerance occur in a spectrum from bothersome at least to life threatening when most severe. Here we discuss nutritional access and its maintenance, introduce and define intolerance, and then review the current literature with regard to principal forms of enteral nutrition intolerance. PMID- 16215144 TI - Monitoring feeding tube placement. AB - The purpose of this literature review is to describe currently available bedside methods to determine feeding tube placement. Described first are methods used at the time of blind insertion to distinguish between gastric and respiratory placement and gastric and small-bowel placement. Discussed next are methods used after feedings are initiated to determine if the tube has remained in the desired position in the gastrointestinal tract. Some of the methods are research-based, whereas others are opinion-based. The level of accuracy of the methods discussed in the review varies widely. No sure non-radiographic method exists to differentiate between respiratory, esophageal, gastric, and small bowel placement of blindly inserted feeding tubes in the fed or unfed state. However, a combination of some of the simpler and more accurate methods may be used to guide feeding tube placement during insertion and help identify the point at which an abdominal radiograph is most likely to confirm the desired location. In addition, methods described in this review can help determine when a radiograph is needed to confirm that a feeding tube has remained in the correct position after the initiation of feedings. Minimizing the number of radiographs taken to assure correct tube placement is important, especially in young children and in the critical care setting where the need for radiographs for other reasons is common. PMID- 16215145 TI - Monitoring enteral nutrition support tolerance in infants and children. AB - Enteral nutrition support is used extensively in the care of infants and children, both for acute and chronic conditions. Monitoring a child's tolerance of enteral feedings is an ongoing challenge. Monitoring routines vary significantly between institutions, practitioners, and patient settings, and a number of definitions are used for "intolerance." Some guidelines have scientific basis and others are passed along in a more anecdotal fashion. This review describes commonly used monitors for tolerance to enteral nutrition for infants and children and discusses pertinent data relevant to practice. PMID- 16215146 TI - Gastric feeding as an extension of an established enteral nutrition protocol. AB - BACKGROUND: Indiscriminate gastric feeding in ICU patients imposes unacceptable risks of aspiration. Believing that a subset of ICU patients can be fed safely via the stomach, we have developed a protocol to identify appropriate patients and guide the bedside clinician in how to safely and effectively feed via the stomach. METHODS: A literature search was done to identify appropriate medical literature. High grade evidence along with local expert opinions were used to develop a protocol. This protocol has been refined and implemented. RESULTS: Based on perceived risk of aspiration, patients are assigned enteral access (ie, stomach vs. distal post-pyloric). Enteral formula is selected based on patient characteristics. It is then advanced by a standard protocol with specific precautions while monitoring for symptoms of intolerance. Management of intolerance is dictated by the type and severity of intolerance. CONCLUSION: We have implemented a gastric feeding into a subset of our ICU patients. Gastric feeding requires certain precautions but appears to be safe. With more experience and better understanding of the pathogenesis gastroparesis, we believe that most ICU patients should be able to safely feed into the stomach. This is logistically easier than post-pyloric feeding and offers physiologic advantages. PMID- 16215147 TI - Factors causing interrupted delivery of enteral nutrition in trauma intensive care unit patients. AB - BACKGROUND: The intent of this study was to ascertain the adequacy of delivery of enteral nutrition (EN) to critically ill adult multiple trauma patients and to identify potential detrimental factors that affect EN delivery. METHODS: Retrospective observational study. Trauma intensive care unit (TICU) in a university-affiliated hospital. Adult patients (>/=18 years of age) admitted to the TICU who received enteral feeding. RESULTS: Fifty-six adult patients were enrolled for study. Patients received, on average, 67% +/- 19% of what was prescribed for 5.7 +/- 2.0 days. A total of 222 occurrences for temporary discontinuation of tube feeding were identified. Gastrointestinal intolerance, as defined by a gastric residual volume of >150 mL, abdominal pain, or >3 liquid stools per day, accounted for only 11% of the occurrences for discontinuation of feeding. Surgery (27%) and diagnostic procedures (15%) represented the majority of reasons for inadequate nutrient delivery. Minor factors for EN interruptions were mechanical feeding tube problems (8%), pharmacy delivery delay (4%), and miscellaneous factors (3%). Multiple and unknown reasons contributed to 14% and 18% of the occurrences, respectively. CONCLUSIONS: Surgery and diagnostic procedures accounted for the largest factor in enteral feeding discontinuations in our critically ill trauma patients. Gastrointestinal intolerance contributed a minor role in the temporary discontinuation of enteral feeding. PMID- 16215148 TI - Benefits of postpyloric enteral access placement by a nutrition support dietitian. AB - BACKGROUND: Although enteral nutrition is considered the preferred strategy for nutrition support, it is often precluded by nasogastric feeding intolerance or the inability to place feeding access into the postpyloric position. In an effort to improve enteral nutrition (EN) outcomes at our institution, the nutrition support dietitian (NSD) began placing postpyloric feeding tubes (PPFT) in intensive care unit patients. METHODS: Intensive care unit patients who received blind, bedside PPFT placements by the NSD (n = 18) were compared with a concurrent age- and diagnosis-matched control group that received standard nutritional care without NSD intervention (n = 18). Interruption of EN infusion, appropriateness of parenteral nutrition (PN) prescription (based on American Society of Parenteral and Enteral Nutrition guidelines), and incidence of ventilator-associated pneumonia (VAP), as defined by the American College of Chest Physicians practice guidelines, were determined in each group. RESULTS: The NSD was successful in positioning the PPFT at or distal to the third portion of the duodenum in 83% of attempts. The PPFT group demonstrated no interruption of enteral feeding compared with 56% in the control group (p < .01) and required 1 (6%) PN initiation in contrast to 8 (44%) in the control group (p < .01). There was a trend toward reduced VAP in the PPFT group (6% vs 28%, p = .07). Of the PN initiations in the control group, 88% were deemed to be potentially avoidable; 6 of 8 PNs were initiated because of gastric residuals. CONCLUSIONS: Enteral nutrition facilitated by NSD placement of postpyloric feeding access is associated with improved tube feeding tolerance and reduced PN use. Further studies are needed to evaluate a possible effect of postpyloric feeding on the incidence of VAP. PMID- 16215149 TI - The impact of implementing an enteral tube feeding protocol on caloric and protein delivery in intensive care unit patients. AB - BACKGROUND: The purpose of this study was to determine the effect of an enteral tube feeding protocol on caloric and protein delivery to intensive care unit (ICU) patients. METHODS: This prospective study consisted of 2 phases: before and after the implementation on an enteral-feeding protocol. The following data were collected: demographics, Acute Physiology and Chronic Health Evaluation II score and Simplified Acute Physiology Score II, caloric and protein requirements, the location of the feeding tube tip, and prokinetic agents use. The primary endpoint was caloric and protein intake as a percentage of the requirement. Secondary endpoints were gastric residuals >150 mL, vomiting episodes, ICU and hospital lengths of stay, mechanical ventilation duration, and ICU and hospital mortality. RESULTS: There were no significant differences between the control (n = 100) and protocol groups (n = 103) in baseline characteristics. The protocol was associated with significant improvement in the 7-day average of caloric intake/requirement (53.9 +/- 2.3% vs 64.5 +/- 2.2%, p = .001) and protein intake/requirement (56.7 +/- 2.6% vs 67.4% +/- 2.7%, p = .005). Caloric and protein intake improved whether the patient was receiving prokinetic agent or not. There was a trend toward lower gastric residual volumes and vomiting episodes in the protocol group. Patients receiving gastric feeding showed significant improvement in caloric intake to levels comparable to patients with postpyloric feeding. CONCLUSIONS: Enteral tube feeding protocol is effective in improving feeding delivery in ICU patients independent of prokinetic agent use. Protocol for enteral tube feeding should be considered in the management of ICU patients, given the positive impact of this nonpharmacologic, non-interventional tool. PMID- 16215150 TI - Increased blood urea nitrogen/creatinine ratio with excess protein. PMID- 16215152 TI - Nutrition and lung disease. PMID- 16215151 TI - Spurious laboratory values from improper sampling of blood containing parenteral nutrition. PMID- 16215153 TI - Nutritional management of the patient with chronic obstructive pulmonary disease. AB - The prevalence and prognostic importance of malnutrition in chronic obstructive pulmonary disease (COPD), the factors that contribute to development of nutritional depletion, and the available evidence regarding effects of nutrition support in these patients are the subjects of this review. Nutritional depletion, as indicated by weight loss and loss of lean body mass, is a common complication of advanced COPD (particularly, but not limited to, the emphysematous type). Low body weight or recent weight loss and in particular depleted lean body mass in patients with COPD have been shown to be independent predictors of mortality, outcomes after acute exacerbations, hospital admission rates, and need for mechanical ventilation. The factors thought to contribute to nutritional depletion in these patients include elevated resting and activity-related energy expenditure, reduced dietary intake relative to resting energy expenditure, accelerated negative nitrogen balance, particularly during acute exacerbations of COPD, medication effects, and perhaps most importantly an elevated systemic inflammatory response. Studies to date suggest that, although it can help limit weight loss and negative energy balance in these patients, the effect of nutritional supplementation alone on clinically significant outcomes such as pulmonary function and exercise capacity is minimal. However, nutritional supplementation may have a role in the management of COPD when provided as part of an integrated rehabilitation program incorporating a structured exercise component as an anabolic stimulus. PMID- 16215154 TI - The use of specialized enteral formulas in pulmonary disease. AB - The relationship between pulmonary disease and nutrition is significant. Malnutrition and its associated adverse pulmonary effects can directly affect outcomes in the individual with pulmonary disease. The use of nutrition support is common for these patients as a supportive or therapeutic measure. Historical reports of adverse respiratory function associated with high parenteral carbohydrate intakes have led to the controversial use of high-fat enteral products in patients with chronic pulmonary dysfunction. The rationale for using this type of formula is that by providing less carbohydrates, carbon dioxide production will be reduced, respiratory quotient will therefore decrease, and a favorable respiratory outcome will result. In the patient with acute respiratory distress syndrome, an inflammatory state exists in which proinflammatory mediators are produced and affect the course of the disease. An enteral formula with modified lipids designed to modulate eicosanoid production and therefore influence the inflammatory cascade is available. This article reviews the rationale for use of modified enteral formulas in both chronic and acute pulmonary disease, reviews the available studies evaluating the efficacy of these formulas, and provides overall recommendations for the use of specialized enteral formulas in individuals with pulmonary disease. PMID- 16215155 TI - The acute respiratory distress syndrome: role of nutritional modulation of inflammation through dietary lipids. AB - The acute respiratory distress syndrome (ARDS) is the most serious form of acute hypoxic respiratory failure. ARDS represents the expression of an acute, diffuse, inflammatory process in the lungs consequent to a variety of infectious and noninfectious conditions. It is characterized pathologically by damage to pulmonary epithelial and endothelial cells, with subsequent alveolar-capillary leak and exudative pulmonary edema. The main clinical features of ARDS include rapid onset of dyspnea, severe defects in gas exchange, and imaging studies demonstrating diffuse pulmonary infiltrates. The role of nutrition in the management of ARDS has traditionally been supportive. Recent research has demonstrated the potential of certain dietary oils (eg, fish oil, borage oil) to modulate pulmonary inflammation, thereby improving lung compliance and oxygenation, and reducing time on mechanical ventilation. This article reviews the alterations in the immune response that underlie ARDS, discusses the physiology of dietary oils as immunonutrients, summarizes animal and human studies that explore the therapeutic effects of dietary oils, and provides clinical recommendations for their use. PMID- 16215156 TI - Nutrition for patients with cystic fibrosis. AB - Cystic fibrosis (CF) is the most frequent, lethal genetic disorder among northern Europeans. The etiology of this autosomal recessive disease is known to be a defect in the cAMP activation of chloride (Cl-) channels in secretory cells in many organs of the body. Although this defect usually leads to severe lung disease, many of these patients also have nutritional deficiencies. Nutrition is one of the key components in the management of CF. Patients are at high risk for malnutrition, which may result in accelerated progression of the disease and increased morbidity. This review will discuss nutrition recommendations for calories, protein, vitamins and minerals, and enteral and parenteral nutrition support practices. PMID- 16215157 TI - Current nutrition management of infants with chronic lung disease. AB - Preterm infants with lung disease present nutrition challenges to health care providers. Malnutrition is common, develops shortly after birth, and may continue into early childhood. Although there are many studies identifying the nutrient deficiencies in infants with chronic lung disease, few randomized trials have explored the effects of nutrition support on the prevention and treatment of chronic lung disease. The purpose of this article is to review current practices and ongoing controversies in the nutrition management of infants with chronic lung disease. PMID- 16215158 TI - Current nutrition practices in adult lung transplantation. AB - Nutrition therapy is vital to the overall management of lung transplant recipients. The objective of this review is to outline the current applications of pre- and posttransplant nutrition management of the adult lung transplant recipient. Pretransplant nutrition therapy decisions are based on cause of end stage lung disease, transplant indications, and pretransplant nutritional status. Maintaining adequate nutrient stores is the major goal of nutrition therapy for patients awaiting transplantation. In the posttransplant course, several gastrointestinal (GI) complications such as gastroesophageal reflux, gastroparesis, and distal intestinal obstruction syndrome complicate nutritional recovery. Long-term nutrition therapy for lung transplant recipients is aimed at management of common comorbid conditions such as obesity, diabetes mellitus, hypertension, osteoporosis, and hyperlipidemia. Lung transplantation outcomes are steadily improving; however, much has yet to be explored to improve the nutrition management of these patients in both the pre- and posttransplantation course. PMID- 16215159 TI - Aspiration and the risk of ventilator-associated pneumonia. AB - Ventilator-associated pneumonia (VAP) is a major concern in the intensive care unit. It is estimated that the risk of developing VAP may be as high as 1% per ventilated day, and the attributable mortality approaches 50% in some series. A growing body of evidence implicates the role of microaspiration of contaminated oropharyngeal and perhaps gastroesophageal secretions into the airways as an integral step in the pathogenesis of VAP. In patients who have been intubated and mechanically ventilated for >72 hours, the majority of VAP is caused by enteric gram-negative organisms, presumably of gastrointestinal origin. As a result, strategies designed to minimize the risk of these contaminated secretions into the normally sterile airways are of paramount importance in terms of VAP prevention. This review highlights the important etiological role of the gut in the development of VAP and also discusses the evidence behind interventions that may modulate the risk of both aspiration and subsequent VAP. PMID- 16215160 TI - Enteral access in mechanically ventilated patients. AB - The early institution of enteral nutrition is now accepted as the preferred route of feeding in critically ill patients with a functioning gastrointestinal tract. It is particularly important to establish early enteral nutrition in mechanically ventilated patients because of the metabolic demands associated with mechanical ventilation. The options for enteral access in mechanically ventilated patients are reviewed, with an emphasis on those techniques that may be performed at the bedside. The advantages, disadvantages, and complications of the different techniques will be considered. PMID- 16215161 TI - The measurement of energy expenditure. AB - Proper nutrition support depends upon the clinician's ability to estimate the patient's energy expenditure. The accuracy of estimation is inversely proportional to the severity of the patient's illness. This fact has spurred academic and industry groups to pursue the measurement of energy expenditure. Harris and Benedict used indirect calorimetry to develop their now-famous equation nearly 100 years ago. The concept of indirect calorimetry is simple; if you know the concentration of inspired gases and expired gases, along with the flow, you can determine the amount of a gas consumed or produced. The complexity and expense of indirect calorimeters suggest that this simple concept is technically challenging. Because we desire to know the energy expenditure of the most critically ill patients, indirect calorimetry is further confounded by the presence of oxygen and mechanical ventilation. This paper will discuss the myriad of variables and obstacles that complicate the measurement of energy expenditure and will suggest methods to avoid or overcome them. PMID- 16215162 TI - Effects of home parenteral nutrition on resting metabolic rate: a case study. AB - Resting metabolic rate (RMR) measurement by indirect calorimetry is often used in acute care; however, indirect calorimetry is rarely performed on patients treated at home with complex therapies such as home parenteral nutrition (HPN). A new hand-held indirect calorimeter is now available to measure RMR and oxygen consumption (VO2). This is a case study of a 35-year-old female who had her RMR and VO2 measured at rest while receiving cyclic HPN and while off of HPN over a period of 8 days. Results indicate that her RMR was higher while receiving HPN. Measuring RMR in patients receiving HPN may be very beneficial to avoid underfeeding and overfeeding. It will be important for the clinician to note if the patient is receiving HPN at the time of the measurement because RMR may be affected by HPN infusion. PMID- 16215163 TI - Determining feeding tube location by gastric and intestinal pH values. AB - BACKGROUND: The purpose of this study was to evaluate a series of pH values of aspirates from feeding tubes to differentiate between gastric and intestinal tube placement. METHODS: One hundred ventilator-supported subjects, with an order for nasoduodenal feeding tube placement were randomly selected. The feeding tube was placed into the stomach, verified by auscultation, and 2 pH measurements were obtained. After the pH measurements were taken, tube location was then verified by fluoroscopy. When the tube was advanced into the small intestine, location was verified by fluoroscopy, and 2 additional pH measurements were obtained. RESULTS: From the selected subjects, both a gastric and intestinal aspirate were obtained from 82 subjects. For the 82 subjects, the mean gastric and intestinal pH values were 4.8 +/- 2.3 and 7.1 +/- 1.0, respectively. For patients receiving acid suppressing agents, these values were 5.0 +/- 2.3 and 7.2 +/- 1.0, respectively. For subjects not receiving acid-suppressing agents, these pH values were 4.0 +/- 2.5 and 6.7 +/- 1.1, respectively. For all patients, using a pH value of 6.5 as a predictor of feeding tube location, the sensitivity and specificity were 0.66 and 0.90. In addition, when an increase in pH > or = 1.0 was used as a predictor of location change for all patients, the sensitivity was 0.58. CONCLUSIONS: The pH value was a reliable predictor of nasoduodenal tube placement. However, radiographic confirmation of location could not be excluded because of the low sensitivity using pH measurements alone. An increase of > or = 1.0 in gastric vs intestinal pH specimens could be useful to determine use of radiography confirmation of tube placement. PMID- 16215164 TI - "Evidence-based" medicine derived from systematic reviews or meta-analyses to develop clinical practice guidelines. PMID- 16215166 TI - Postnatal developmental consequences of altered insulin sensitivity in female sheep treated prenatally with testosterone. AB - Prenatally testosterone (T)-treated female sheep exhibit ovarian and endocrinological features that resemble those of women with polycystic ovarian syndrome (PCOS), which include luteinizing hormone excess, polyfollicular ovaries, functional hyperandrogenism, and anovulation. In this study, we determined the developmental impact of prenatal T treatment on insulin sensitivity indexes (ISI), a variable that is affected in a majority of PCOS women. Pregnant ewes were treated with 60 mg testosterone propionate intramuscularly in cottonseed oil two times a week or vehicle [control (C)] from days 30 to 90 of gestation. T-females weighed less than C-females or males (P < 0.05) at birth and at 5 wk of age. T-females had an increased anogenital ratio. An intravenous glucose tolerance test followed by an insulin tolerance test conducted after an overnight fast at 5, 20, and 30 wk of age (n = 7-8/group) revealed that ISI were higher at 5 than 30 wk of age in C-females, reflective of a developing insulin resistance associated with puberty. T-females had higher basal insulin levels, higher fasting insulin-to-glucose ratio, and higher incremental area under the insulin curve to the glucose challenge. The ISI of T females was similar to that of males. No differences in ISI were evident between groups at 20 and 30 wk of age. Mean basal plasma glucose concentrations and glucose disappearance and uptake did not differ between groups at any age. Our findings suggest that prenatal T treatment leads to offspring with reduced birth weight and impaired insulin sensitivity in early postnatal life. PMID- 16215167 TI - Photoreceptor for curling behavior in Peranema trichophorum and evolution of eukaryotic rhodopsins. AB - When it is gliding, the unicellular euglenoid Peranema trichophorum uses activation of the photoreceptor rhodopsin to control the probability of its curling behavior. From the curled state, the cell takes off in a new direction. In a similar manner, archaea such as Halobacterium use light activation of bacterio- and sensory rhodopsins to control the probability of reversal of the rotation direction of flagella. Each reversal causes the cell to change its direction. In neither case does the cell track light, as known for the rhodopsin dependent eukaryotic phototaxis of fungi, green algae, cryptomonads, dinoflagellates, and animal larvae. Rhodopsin was identified in Peranema by its native action spectrum (peak at 2.43 eV or 510 nm) and by the shifted spectrum (peak at 3.73 eV or 332 nm) upon replacement of the native chromophore with the retinal analog n-hexenal. The in vivo physiological activity of n-hexenal incorporated to become a chromophore also demonstrates that charge redistribution of a short asymmetric chromophore is sufficient for receptor activation and that the following isomerization step is probably not required when the rest of the native chromophore is missing. This property seems universal among the Euglenozoa, Plant, and Fungus kingdom rhodopsins. The rhodopsins of animals have yet to be studied in this respect. The photoresponse appears to be mediated by Ca2+ influx. PMID- 16215165 TI - Is insulin an anabolic agent in bone? Dissecting the diabetic bone for clues. AB - Diabetic osteoporosis is increasingly recognized as a significant comorbidity of type 1 diabetes mellitus. In contrast, type 2 diabetes mellitus is more commonly associated with modest increases in bone mineral density for age. Despite this dichotomy, clinical, in vivo, and in vitro data uniformly support the concept that new bone formation as well as bone microarchitectural integrity are altered in the diabetic state, leading to an increased risk for fragility fracture and inadequate bone regeneration following injury. In this review, we examine the contribution that insulin, as a potential anabolic agent in bone, may make to the pathophysiology of diabetic bone disease. Specifically, we have assimilated human and animal data examining the effects of endogenous insulin production, exogenous insulin administration, insulin sensitivity, and insulin signaling on bone. In so doing, we present evidence that insulin, acting as an anabolic agent in bone, can preserve and increase bone density and bone strength, presumably through direct and/or indirect effects on bone formation. PMID- 16215168 TI - Analysis of Paramecium tetraurelia A-51 surface antigen gene mutants reveals positive-feedback mechanisms for maintenance of expression and temperature induced activation. AB - In Paramecium tetraurelia, variable surface antigen loci show mutually exclusive expression which is controlled primarily at the transcriptional level. Clonally stable expression of a single antigen has attracted models involving self regulation by their gene products. However, direct demonstration of self-feedback at the molecular level has been complicated due to the inability to separate the functional gene from its product as well as copy number effects associated with injected extrachromosomal DNA in the polygenomic somatic nucleus. In this study, we exploited several germ line termination and frameshift mutations in the A-51 surface antigen gene to analyze variable surface antigen expression. These mutant alleles have the same copy number as the wild-type allele and therefore eliminate possible copy number effects. The mutant alleles were not transcribed at 27 degrees C, consistent with positive-feedback models for gene expression. However, further analysis showed that high temperatures (34 degrees C) induced transcription of the mutant A genes even in the presence of a different antigen on the cell surface. Thus, transcription was temperature dependent. Unlike wild type cells, transcription of the mutant A genes at high temperatures was not maintained after temperature shift back to 27 degrees C in homozygous mutant cells. Importantly, transcription of the mutant allele was maintained at 27 degrees C in heterozygous cells with one copy of the wild-type allele. These results indicate that expression of the wild-type gene is required to stabilize its own transcriptional state at 27 degrees C. PMID- 16215169 TI - Mg-protoporphyrin IX and heme control HEMA, the gene encoding the first specific step of tetrapyrrole biosynthesis, in Chlamydomonas reinhardtii. AB - HEMA encodes glutamyl-tRNA reductase (GluTR), which catalyzes the first step specific for tetrapyrrole biosynthesis in plants, archaea, and most eubacteria. In higher plants, GluTR is feedback inhibited by heme and intermediates of chlorophyll biosynthesis. It plays a key role in controlling flux through the tetrapyrrole biosynthetic pathway. This enzyme, which in Chlamydomonas reinhardtii is encoded by a single gene (HEMA), exhibits homology to GluTRs of higher plants and cyanobacteria. HEMA mRNA accumulation was inducible not only by light but also by treatment of dark-adapted cells with Mg-protoporphyrin IX (MgProto) or hemin. The specificity of these tetrapyrroles as inducers was demonstrated by the absence of induction observed upon the feeding of protoporphyrin IX, the precursor of both heme and MgProto, or chlorophyllide. The HEMA mRNA accumulation following treatment of cells with light and hemin was accompanied by increased amounts of GluTR. However, the feeding of MgProto did not suggest a role for Mg-tetrapyrroles in posttranscriptional regulation. The induction by light but not that by the tetrapyrroles was prevented by inhibition of cytoplasmic protein synthesis. Since MgProto is synthesized exclusively in plastids and heme is synthesized in plastids and mitochondria, the data suggest a role of these compounds as organellar signals that control expression of the nuclear HEMA gene. PMID- 16215170 TI - Comparative gene genealogies indicate that two clonal lineages of Cryptococcus gattii in British Columbia resemble strains from other geographical areas. AB - Cryptococcus gattii has recently emerged as a pathogen of humans and animals in the temperate climate of Vancouver Island, British Columbia (B.C.). The majority (approximately 95%) of the isolates from the island belong to the VGII molecular type, and the remainder belong to the VGI molecular type. The goals of this study were to compare patterns of molecular variation among C. gattii isolates from B.C. with those from different areas of the world and to investigate the population structure using a comparative gene genealogy approach. Our results indicate that the C. gattii population in B.C. comprises at least two divergent lineages, corresponding to previously identified VGI and VGII molecular types. The genealogical analysis of strains suggested a predominantly clonal population structure among B.C. isolates, while there was evidence for sexual recombination between different molecular types on a global scale. We found no geographic pattern of strain relationships, and nucleotide sequence comparisons revealed that genotypes among isolates from B.C. were also present among isolates from other areas of the world, indicating extensive strain dispersal. The nucleotide sequence diversity among isolates from B.C. was similar to that among isolates from other areas of the world. PMID- 16215171 TI - Rho4 GTPase is involved in secretion of glucanases during fission yeast cytokinesis. AB - Rho GTPases are regulators of signaling pathways that control actin organization and cell polarity processes in all eukaryotic cells. In Schizosaccharomyces pombe, Rho4p is involved in the regulation of septum degradation during cytokinesis. Here we show that Rho4p participates in the secretion of the glucanases Eng1p and Agn1p, which are responsible for the septum degradation. First, eng1+ or agn1+ overexpression suppressed the rho4delta multiseptation phenotype, and simultaneous overproduction of Rho4p and Eng1p or of Rho4p and Agn1p caused a dramatic lysis. Second, Rho4p was not necessary for Eng1p-mediated glucanase activity as measured in cell extracts; however, rho4delta cells have a lower level of (1,3)-beta-D-glucanase activity in the culture medium. Additionally, Eng1- or Agn1-green fluorescent protein did not properly localize to the septum in rho4delta cells grown at 37 degrees C. There was a decreased amount of these enzymes in the cell wall and in the culture medium of rho4delta cells at 37 degrees C. These results provide evidence that Rho4p is involved in the regulation of Eng1p and Agn1p secretion during cytokinesis. PMID- 16215172 TI - Nuclear accumulation of the GATA factor AreA in response to complete nitrogen starvation by regulation of nuclear export. AB - Both the availability and the quality of nutrients affect cellular functions by controlling gene activity. AreA, a member of the GATA family of transcription factors, globally activates expression of genes involved in nitrogen source utilization in Aspergillus nidulans. The quality of the nitrogen source determines the level and activation capacity of AreA through controls at the level of areA mRNA stability and by interaction of AreA with the corepressor NmrA. The availability of potential nitrogen sources also affects the activation capacity of AreA. We show that the complete absence of a nitrogen source results in an enhanced level of AreA-dependent gene expression and that this response is independent of mechanisms regulating AreA activity in response to nitrogen source quality. During nitrogen starvation AreA accumulates in the nucleus, but the presence of a potential nitrogen source or carbon starvation prevents this accumulation. Furthermore, accumulated AreA is rapidly lost from the nuclei of nitrogen-starved cells when a nitrogen source is supplied or when a carbon source is absent, and this accompanies arrest of the AreA-dependent nitrogen starvation response on regulated gene expression. By the generation of a leptomycin B sensitive mutant, we have been able to show that nuclear exit occurs via the CrmA exportin. We conclude that sensing mechanisms discriminate between starvation and the presence of potential nutrients that can signal to the AreA transcription factor. Nitrogen source availability, but not quality, affects nuclear accumulation by regulating nuclear exit of AreA, providing a rapid response to changes in the supply of nutrients. PMID- 16215173 TI - Candida albicans-conditioned medium protects yeast cells from oxidative stress: a possible link between quorum sensing and oxidative stress resistance. AB - Candida albicans, the most frequent fungal pathogen of humans, encounters high levels of oxidants following ingestion by professional phagocytes and through contact with hydrogen peroxide-producing bacteria. In this study, we provide evidence that C. albicans is able to coordinately regulate the oxidative stress response at the global cell population level by releasing protective molecules into the surrounding medium. We demonstrate that conditioned medium, which is defined as a filter-sterilized supernatant from a C. albicans stationary-phase culture, is able to protect yeast cells from both hydrogen peroxide and superoxide anion-generating agents. Exponential-phase yeast cells preexposed to conditioned medium were able to survive levels of oxidative stress that would normally kill actively growing yeast cells. Heat treatment, digestion with proteinase K, pH adjustment, or the addition of the oxidant scavenger alpha tocopherol did not alter the ability of conditioned medium to induce a protective response. Farnesol, a heat-stable quorum-sensing molecule (QSM) that is insensitive to proteolytic enzymes and is unaffected by pH extremes, is partly responsible for this protective response. In contrast, the QSM tyrosol did not alter the sensitivity of C. albicans cells to oxidants. Relative reverse transcription-PCR analysis indicates that Candida-conditioned growth medium induces the expression of CAT1, SOD1, SOD2, and SOD4, suggesting that protection may be mediated through the transcriptional regulation of antioxidant-encoding genes. Together, these data suggest a link between the quorum-sensing molecule farnesol and the oxidative stress response in C. albicans. PMID- 16215174 TI - Novel regulatory function for the CCAAT-binding factor in Candida albicans. AB - Candida albicans is an opportunistic human pathogen that can sense environmental changes and respond by altering its cell morphology and physiology. A number of environmental factors have been shown to influence this dimorphic transition, including pH, starvation, serum, and amino acids. In this report, we investigate the function of the C. albicans CCAAT-binding factor. In Saccharomyces cerevisiae, this heterooligomeric transcriptional activator stimulates the expression of genes that encode proteins involved in respiration. To examine the function of this transcription factor in C. albicans, we cloned CaHAP5 and generated a hap5delta/hap5delta mutant of C. albicans. Using mobility shift studies, we identified four separate complexes from C. albicans cell extracts whose DNA-binding activities were abolished in the hap5delta/hap5delta mutant, suggesting that they represented sequence-specific CCAAT-binding complexes. We found that the C. albicans hap5delta homozygote was defective in hyphal development under a variety of conditions, and the mutant displayed a carbon source-dependent "hyperfilamentation" phenotype under certain growth conditions. In addition, the mRNA levels for two enzymes involved in respiration, encoded by COX5 and CYC1, were overexpressed in the hap5delta/hap5delta mutant when grown in medium containing amino acids as the sole carbon and nitrogen source. Thus, the C. albicans CCAAT-binding factor appeared to function as a repressor of genes encoding mitochondrial electron transport components, in contrast to its activator function in S. cerevisiae. These data provide the first evidence that the CCAAT-binding factor can act as a transcriptional repressor and raise new and interesting questions about how carbon metabolism is regulated in this opportunistic human pathogen. PMID- 16215175 TI - The Candida albicans vacuole is required for differentiation and efficient macrophage killing. AB - Yeast-hypha differentiation is believed to be necessary for the normal progression of Candida albicans infections. The emergence and extension of a germ tube from a parental yeast cell are accompanied by dynamic changes in vacuole size and morphology. Although vacuolar function is required during this process, it is unclear if it is vacuolar expansion or some other vacuolar function that is important. We previously described a C. albicans vps11Delta mutant which lacked a recognizable vacuole compartment and with defects in multiple vacuolar functions. These include sensitivities to stress, reduced proteolytic activities, and severe defects in filamentation. Herein we utilize a partially functional VPS11 allele (vps11hr) to help define which vacuolar functions are required for differentiation and which influence interaction with macrophages. Mutant strains harboring this allele are not osmotically or temperature sensitive and have normal levels of secreted aspartyl protease and carboxypeptidase Y activity but have a fragmented vacuole morphology. Moreover, this mutant is defective in filamentation, suggesting that the major role the vacuole plays in yeast-hypha differentiation may relate directly to its morphology. The results of this study support the hypothesis that vacuole expansion is required during germ tube emergence. Both vps11 mutants were severely attenuated in their ability to kill a macrophage cell line. The viability of the vps11delta mutant was significantly reduced during macrophage interaction compared to that in the control strains, while the vps11hr mutant was unaffected. This implies some vacuolar functions are required for Candida survival within the macrophage, while additional vacuolar functions are required to inflict injury on the macrophage. PMID- 16215177 TI - Microtubules in Candida albicans hyphae drive nuclear dynamics and connect cell cycle progression to morphogenesis. AB - Candida albicans is an opportunistic fungal pathogen whose virulence is related to its ability to switch between yeast, pseudohyphal, and true-hyphal morphologies. To ask how long-distance nuclear migration occurs in C. albicans hyphae, we identified the fundamental properties of nuclear movements and microtubule dynamics using time-lapse microscopy. In hyphae, nuclei migrate to, and divide across, the presumptive site of septation, which forms 10 to 15 microm distal to the basal cell. The mother nucleus returns to the basal cell, while the daughter nucleus reiterates the process. We used time-lapse microscopy to identify the mechanisms by which C. albicans nuclei move over long distances and are coordinated with hyphal morphology. We followed nuclear migration and spindle dynamics, as well as the time and position of septum specification, defined it as the presumptum, and established a chronology of nuclear, spindle, and morphological events. Analysis of microtubule dynamics revealed that premitotic forward nuclear migration is due to the repetitive sliding of astral microtubules along the cell cortex but that postmitotic forward and reverse nuclear migrations are due primarily to spindle elongation. Free microtubules exhibit cell cycle regulation; they are present during interphase and disappear at the time of spindle assembly. Finally, a growth defect in strains expressing Tub2-green fluorescent protein revealed a connection between hyphal elongation and the nuclear cell cycle that is coordinated by hyphal length and/or volume. PMID- 16215176 TI - Global role of the protein kinase Gcn2 in the human pathogen Candida albicans. AB - The pathogen Candida albicans responds to amino acid starvation by activating pseudohyphal development and the expression of amino acid biosynthetic genes (GCN response). In Saccharomyces cerevisiae, the GCN response is dependent on Gcn2, which regulates the translation of the transcription factor Gcn4. Therefore, we examined the role of Gcn2 in C. albicans by using molecular, cellular, and genomic approaches. We show that C. albicans GCN2 encodes an eIF2alpha kinase, like its S. cerevisiae homologue. However, GCN4 appears to be regulated mainly at the transcriptional level in C. albicans. Furthermore, the inactivation of C. albicans Gcn2 only partially attenuates growth under amino acid starvation conditions and resistance to the histidine analogue 3-aminotriazole. Our comparison of the Gcn4 and Gcn2 regulons by transcript profiling reinforces the view that Gcn2 contributes to, but is not essential for, the activation of general amino acid control in C. albicans. PMID- 16215178 TI - Ras1-induced hyphal development in Candida albicans requires the formin Bni1. AB - Formins are downstream effector proteins of Rho-type GTPases and are involved in the organization of the actin cytoskeleton and actin cable assembly at sites of polarized cell growth. Here we show using in vivo time-lapse microscopy that deletion of the Candida albicans formin homolog BNI1 results in polarity defects during yeast growth and hyphal stages. Deletion of the second C. albicans formin, BNR1, resulted in elongated yeast cells with cell separation defects but did not interfere with the ability of bnr1 cells to initiate and maintain polarized hyphal growth. Yeast bni1 cells were swollen, showed an increased random budding pattern, and had a severe defect in cytokinesis, with enlarged bud necks. Induction of hyphal development in bni1 cells resulted in germ tube formation but was halted at the step of polarity maintenance. Bni1-green fluorescent protein is found persistently at the hyphal tip and colocalizes with a structure resembling the Spitzenkorper of true filamentous fungi. Introduction of constitutively active ras1G13V in the bni1 strain or addition of cyclic AMP to the growth medium did not bypass bni1 hyphal growth defects. Similarly, these agents were not able to suppress hyphal growth defects in the wal1 mutant which is lacking the Wiskott Aldrich syndrome protein (WASP) homolog. These results suggest that the maintenance of polarized hyphal growth in C. albicans requires coordinated regulation of two actin cytoskeletal pathways, including formin-mediated secretion and WASP-dependent endocytosis. PMID- 16215179 TI - DNA-bound Bas1 recruits Pho2 to activate ADE genes in Saccharomyces cerevisiae. AB - Expression of the genes in the ADE regulon of Saccharomyces cerevisiae is repressed by the presence of purine bases in the extracellular medium and derepressed when cells are grown in the absence of purines. Derepression requires the transcriptional activators Bas1 and Pho2, as well as the biosynthetic intermediates 5'-phosphoribosyl-4-succinocarboxamide-5-aminoimidazole (SAICAR) and 5'-phosphoribosyl-4-carboxamide- 5-aminoimidazole (AICAR). In this study, we investigated if nuclear localization and binding to promoter DNA by the activators are regulated by purines. Using indirect immunofluorescence, we found that Bas1 is localized to the nucleus under both repressing and derepressing conditions. Importantly, we detected Bas1 bound to promoter DNA under both conditions using chromatin immunoprecipitation assays at several ADE promoters (ADE1, ADE2, ADE4, and ADE5,7) and HIS4. We analyzed the binding of Bas1 to wild type and mutant sequences of the ADE5,7 promoters in vivo, and found that Bas1 binds independently to each of its two binding sites. Pho2 was not required for the association of Bas1 with chromosomal DNA, but it was required for an increase in Bas1-immunoprecipitated DNA. The presence of Pho2 at promoters was dependent on Bas1 and occurred only under derepressing conditions when the ADE genes are transcribed at elevated levels. We propose a model for regulation of the ADE genes in which DNA-bound Bas1 is inactive due to masking of its activation domain and Pho2 binds poorly to promoters when cells have sufficient purine nucleotides. Upon limitation for purines, the SAICAR/AICAR regulatory signal is transmitted to the nucleus to increase Bas1 and Pho2 interaction, recruiting Pho2 to promoters and freeing the activation domains for transactivation. PMID- 16215180 TI - HdaA, a major class 2 histone deacetylase of Aspergillus nidulans, affects growth under conditions of oxidative stress. AB - Histone deacetylases (HDACs) catalyze the removal of acetyl groups from the epsilon-amino group of distinct lysine residues in the amino-terminal tail of core histones. Since the acetylation status of core histones plays a crucial role in fundamental processes in eukaryotic organisms, such as replication and regulation of transcription, recent research has focused on the enzymes responsible for the acetylation/deacetylation of core histones. Very recently, we showed that HdaA, a member of the Saccharomyces cerevisiae HDA1-type histone deacetylases, is a substantial contributor to total HDAC activity in the filamentous fungus Aspergillus nidulans. Now we demonstrate that deletion of the hdaA gene indeed results in the loss of the main activity peak and in a dramatic reduction of total HDAC activity. In contrast to its orthologs in yeast and higher eukaryotes, HdaA has strong intrinsic activity as a protein monomer when expressed as a recombinant protein in a prokaryotic expression system. In vivo, HdaA is involved in the regulation of enzymes which are of vital importance for the cellular antioxidant response in A. nidulans. Consequently, deltahdaA strains exhibit significantly reduced growth on substrates whose catabolism generates molecules responsible for oxidative stress conditions in the fungus. Our analysis revealed that reduced expression of the fungal catalase CatB is jointly responsible for the significant growth reduction of the hdaA mutant strains. PMID- 16215181 TI - Chlamydospore formation during hyphal growth in Cryptococcus neoformans. AB - Cryptococcus neoformans, a basidiomycetous fungal pathogen, infects hosts through inhalation and can cause fatal meningoencephalitis in individuals if untreated. This fungus undergoes a dimorphic transition from yeast to filamentous growth during mating and monokaryotic fruiting, which leads to the production of hyphae and airborne infectious basidiospores. Here we characterized a novel morphological feature associated with the filamentous stages of the life cycle of C. neoformans which resembles resting or survival structures known as chlamydospores in other fungi. The C. neoformans chlamydospore-like structure is rich in glycogen, suggesting that it might have a role as an energy store. However, characterization of mutants with decreased or increased levels of glycogen production showed that glycogen levels have little effect on filamentous growth, sporulation, or chlamydospore formation. These results suggest that the formation of chlamydospores is independent of glycogen accumulation level. We also show that chlamydospore formation does not require successful sporulation and that the presence of chlamydospores is not sufficient for sporulation. Although the biological functions of chlamydospores remain to be established for this pathogenic fungus, their formation appears to be an integral part of the filamentation process, suggesting that they could be necessary to support sexual sporulation under adverse conditions and thereby facilitate the production of infectious basidiospores or long-term survival propagules in harsh environments. PMID- 16215183 TI - The loss of federal support for multisite demonstrations. PMID- 16215182 TI - Prespore cell fate bias in G1 phase of the cell cycle in Dictyostelium discoideum. AB - By generating a population of Dictyostelium cells that are in the G1 phase of the cell cycle we have examined the influence of cell cycle status on cell fate specification, cell type proportioning and its regulation, and terminal differentiation. The lack of observable mitosis during the development of these cells and the quantification of their cellular DNA content suggests that they remain in G1 throughout development. Furthermore, chromosomal DNA synthesis was not detectable these cells, indicating that no synthesis phase had occurred, although substantial mitochondrial DNA synthesis did occur in prespore cells. The G1-phase cells underwent normal morphological development and sporulation but displayed an elevated prespore/prestalk ratio of 5.7 compared to the 3.0 (or 3:1) ratio normally observed in populations dominated by G2-phase cells. When migrating slugs produced by G1-phase cells were bisected, each half could reestablish the 5.7 (or 5.7:1) prespore/prestalk ratio. These results demonstrate that Dictyostelium cells can carry out the entire developmental cycle in the G1 phase of the cell cycle and that passage from G2 into G1 phase is not required for sporulation. Our results also suggest that the population asymmetry provided by the distribution of cells around the cell cycle at the time of starvation is not strictly required for cell type proportioning. Finally, when developed together with G2-phase cells, G1-phase cells preferentially become prespore cells and exclude G2-phase cells from the prespore-spore cell population, suggesting that G1-phase cells have an advantage over G2-phase cells in executing the spore cell differentiation pathway. PMID- 16215184 TI - Economic grand rounds: improving the quality of depression care in Medicaid. PMID- 16215185 TI - Personal accounts: "doctor's standing orders". PMID- 16215186 TI - Twelve-month outcomes of trauma-informed interventions for women with co occurring disorders. AB - OBJECTIVE: Women with co-occurring mental health and substance use disorders frequently have a history of interpersonal violence, and past research has suggested that they are not served effectively by the current service system. The goal of the Women, Co-occurring Disorders, and Violence Study was to develop and test the effectiveness of new service approaches specifically designed for these women. METHODS: A quasi-experimental treatment outcome study was conducted from 2001 to 2003 at nine sites. Although intervention specifics such as treatment length and modality varied across sites, each site used a comprehensive, integrated, trauma-informed, and consumer-involved approach to treatment. Substance use problem severity, mental health symptoms, and trauma symptoms were measured at baseline, and follow-up data were analyzed with prospective meta analysis and hierarchical linear modeling. RESULTS: A total of 2,026 women had data at the 12-month follow-up: 1,018 in the intervention group and 1,008 in the usual-care group. For substance use outcomes, no effect was found. The meta analysis demonstrated small but statistically significant overall improvement in women's trauma and mental health symptoms in the intervention relative to the usual-care comparison condition. Analysis of key program elements demonstrated that integrating substance abuse, mental health, and trauma-related issues into counseling yielded greater improvement, whereas the delivery of numerous core services yielded less improvement relative to the comparison group. A few person level characteristics were associated with increases or decreases in the intervention effect. These neither moderated nor supplanted the effects of integrated counseling. CONCLUSIONS: Outcomes for women with co-occurring disorders and a history of violence and trauma may improve with integrated treatment. PMID- 16215187 TI - Service use and costs for women with co-occurring mental and substance use disorders and a history of violence. AB - OBJECTIVES: This study examined the 12-month cost of the array of services used by women with co-occurring mental health and substance use disorders and a history of violence and trauma who participated in the Women, Co-occurring Disorders, and Violence Study (WCDVS). The study compared costs of the intervention and external services for women in the WCDVS intervention in outpatient and residential settings-which provided comprehensive, integrated, and trauma-informed services-with the costs for women in the usual-care comparison group. The study also compared costs with recorded clinical outcomes. METHODS: Costs of service use were examined for 2,026 women who participated in the WCDVS (N=1,018) and in the comparison group (N=1,008). Women were interviewed three, six, nine, and 12 months after baseline about any service use in the past three months. Costs for these services, along with indirect costs (participants' time and transportation) were estimated by using a variety of sources. A number of cost estimates were analyzed by using either ordinary least squares regression or two-part models. RESULTS: The average participant had almost 43,000 dollars in costs related to their service use during the 12 months after baseline. Women in the intervention group had lower service costs and higher overall costs than those in the comparison group, but the null hypotheses of no difference in any cost measure between groups was not rejected. Also, the null hypothesis of no difference in the probability of accessing services external to the study intervention was not rejected. CONCLUSIONS: Because no differences were detected in costs but improvements were seen in clinical outcomes, the interventions offered in the WCDVS may be more efficient than usual care. PMID- 16215188 TI - Design strengths and issues of SAMHSA's Women, Co-occurring Disorders, and Violence Study. AB - In 1998 the Substance Abuse and Mental Health Services Administration launched the Women, Co-occurring Disorders, and Violence Study (WCDVS). The WCDVS developed, implemented, and evaluated the outcomes and costs of comprehensive, trauma-informed treatment programs for women with a history of violence and trauma who have substance use and mental health disorders. This article discusses the overall design features of the study, issues related to such a design, results of the outcomes and cost evaluations, and suggestions for future research. The nine WCDVS sites were located across the continental United States, with six sites on the East Coast, two on the West Coast, and one in Colorado. A total of 2,729 women (1,415 in the intervention condition and 1,314 in the comparison condition) were enrolled over the 13.5-month baseline accrual period (January 2001 through February 2002). Follow-up interviews were conducted with all participants at three, six, nine and 12 months post-baseline. Women in the intervention and the comparison groups showed improvement in outcomes in four areas: alcohol use, drug use, mental health, and trauma. At six months women in the intervention group scored modestly better than women in the usual-care group for outcome measures for drug use, trauma, and mental health. At 12 months women in the intervention group maintained their improvement in drug use outcomes and continued to improve in mental health and trauma outcomes. After a start-up period, costs for services were not significantly different between the intervention and comparison groups at both follow-up points. Despite their very modest nature, the WCDVS results are promising. PMID- 16215189 TI - Effects of job development and job support on competitive employment of persons with severe mental illness. AB - OBJECTIVES: Few studies have sought to determine which specific supported employment services improve employment outcomes for people with pyschiatric disabilities. This study examined the effects of job development and job support among other services on acquisition and retention of competitive employment. METHODS: Data used in the analysis came from seven sites of the Employment Intervention Demonstration Program. Employment data were collected weekly for a period up to 24 months for 1,340 participants. A random-effects meta-analysis was conducted. RESULTS: Job development increased the probability of obtaining competitive employment. The effects of job development on job acquisition remained after the effects of other factors were controlled for. Job support was associated with more months in the first competitive job but not total hours worked. However, no evidence for the causal role of job support was found in analyses that tested the effects of job support after the job support was provided. The causal role of job support alone was also cast in doubt by the fact that a substantial overlap existed between individuals who received job support and vocational counseling. CONCLUSIONS: Job development is a very effective service when the goal is job acquisition. Job support is associated with retention of a first competitive job, but its causal role is questionable. PMID- 16215190 TI - Assessment of Medicaid managed behavioral health care for persons with serious mental illness. AB - OBJECTIVES: This five-site study compared Medicaid managed behavioral health programs and fee-for-service programs on use and quality of services, satisfaction, and symptoms and functioning of adults with serious mental illness. METHODS: Adults with serious mental illness in managed care programs (N=958) and fee-for-service programs (N=1,011) in five states were interviewed after the implementation of managed care and six months later. After a multiple regression to standardize the groups for case mix differences, a meta-analysis using a random-effects model was conducted, and bioequivalence methods were used to determine whether differences were significant for clinical or policy purposes. RESULTS: A significantly smaller proportion of the managed care group received inpatient care (5.7 percent compared with 11.5 percent). The managed care group received significantly more hours of primary care (4.9 compared with 4.5 hours) and was significantly less healthy. However, none of these differences exceed the bioequivalence criterion of 5 percent. Managed care and fee for service were "not different but not equivalent" on 20 of 34 dependent variables. Cochrane's Q statistic, which measured intersite consistency, was significant for 20 variables. CONCLUSIONS: Managed care and fee-for-service Medicaid programs did not differ on most measures; however, a lack of sufficient power was evident for many measures. Full endorsement of managed care for vulnerable populations will require further research that assumes low penetration rates and intersite variability. PMID- 16215191 TI - The effectiveness of skills training for improving outcomes in supported employment. AB - OBJECTIVE: This study evaluated whether a supplementary skills training program improved work outcomes for clients enrolled in supported employment programs. METHODS: Thirty-five recently employed clients with severe mental illness who were receiving supported employment services at a free-standing agency were randomly assigned to participate in either the workplace fundamentals program, a skills training program designed to make work more "successful and satisfying," or treatment as usual. Knowledge of workplace fundamentals (for example, identifying workplace stressors, problem solving, and improving job performance) was assessed at baseline and at nine months; employment outcomes and use of additional vocational services were tracked for 18 months. RESULTS: Clients in the workplace fundamentals group (N=17) improved more in knowledge of workplace fundamentals than those in the control group (N=18) at the nine-month follow-up, but the two groups did not differ in the number of hours or days worked, salary earned, or receipt of additional vocational services over the 18-month period. In general, clients in this study had higher educational levels and better employment outcomes than clients in most previous studies of supported employment, making it difficult to detect possible effects of the skills training intervention on work. CONCLUSIONS: Supplementary skills training did not improve work outcomes for clients who were receiving supported employment. PMID- 16215192 TI - Generalizability of studies on mental health treatment and outcomes, 1981 to 1996. AB - OBJECTIVE: This study operationalized and measured the external validity, or generalizability, of studies on mental health treatment and outcomes published in four journals between 1981 and 1996. METHOD: MEDLINE was searched for articles on mental health treatment and outcomes that were published in four leading psychiatry and psychology journals between 1981 and 1996. A 156-item instrument was used to assess generalizability of study findings. RESULTS: Of more than 9,000 citations, 414 eligible studies were identified. Inclusion of community sites and patients from racial or ethnic minority groups were documented in only 12 and 25 percent of studies, respectively. Random or systematic sampling methods were rare (3 percent), and 75 percent of studies did not explicitly address sample representativeness. Studies with funding from the National Institute of Mental Health (NIMH) were more likely than those without NIMH funding to document the inclusion of patients from minority groups (30 percent compared with 20 percent). Randomized studies were more likely than nonrandomized studies to document the inclusion of patients from minority groups (28 percent compared with 17 percent), include patients with comorbid psychiatric conditions (31 percent compared with 19 percent), and attend to sample representativeness (28 percent compared with 15 percent). Modest improvements were seen over time in inclusion of patients from minority groups, inclusion of patients with psychiatric comorbidities, and attention to sample representativeness. CONCLUSIONS: Generalizability of studies on treatments and outcomes, whether experimental or observational, remained low and poorly documented over the 16-year period. PMID- 16215194 TI - Relapse of substance use disorder and its prevention among persons with co occurring disorders. AB - This article summarizes the scientific literature on the relapse process, describes the basic principles of relapse prevention treatment, highlights the major empirical studies, and offers suggestions for future research and application, especially in terms of ongoing care for persons with co-occurring disorders. Relapse prevention treatments have a well-established efficacy and effectiveness for persons with substance use disorders. Key ingredients include reducing exposure to substances, fostering motivation for abstinence, self monitoring, recognizing and coping with cravings and negative affect, identifying thought processes with relapse potential, and deploying, if necessary, a crisis plan. Relapse prevention approaches may be best suited for persons in the action of maintenance stages of treatment or recovery. Further research is needed to examine relapse prevention therapies as a key component to continuing care for persons with co-occurring substance use and psychiatric disorders. PMID- 16215195 TI - Substance abuse relapse and factors associated with relapse in an inner-city sample of patients with dual diagnoses. AB - OBJECTIVE: This study documented rates of substance abuse relapse and explored factors associated with sustained remission among consumers with severe mental illness in a large, urban clinical sample. METHODS: Existing clinical records of consumers with severe mental illness and co-occurring substance use disorders who had achieved remission and who were interviewed at two or more subsequent follow up points (12 months after remission) were reviewed. Consumers who relapsed within 12 months after remission were compared with those who maintained remission on demographic, clinical, and functional indicators. RESULTS: Of the 133 consumers who achieved remission, 91 (68 percent) had maintained remission at six-month follow-up, and 69 (52 percent) had maintained remission at 12-month follow-up. The strongest factors associated with maintenance of remission at 12 months were older age and living in Thresholds residential programs. Multivariate analysis showed that consumers who were older, held jobs, and lived in Thresholds residential programs at initial remission had a higher likelihood of maintaining remission at 12 months. To explore the potential impact of program dropout on the results, supplemental analyses using a third group without 12-month follow-up data were conducted. These analyses indicated that program dropouts were younger and less likely to live in Thresholds residential programs at initial remission. CONCLUSIONS: Age, therapeutic residential programming, and, to a lesser degree, employment appear to be potential factors to consider in the development of relapse prevention models. PMID- 16215196 TI - Substance abuse relapse in a ten-year prospective follow-up of clients with mental and substance use disorders. AB - OBJECTIVES: This study addressed the rate and predictors of substance abuse relapse among clients with severe mental illness who had attained full remission from substance abuse. METHODS: In a ten-year prospective follow-up study of clients with co-occurring severe mental and substance use disorders, 169 clients who had attained full remission, defined according to DSM-III-R as at least six months without evidence of abuse or dependence, were identified. The Kaplan-Meier survival curve was developed to show the pattern of relapse, and a discrete-time survival analysis was used to identify predictors of relapse. RESULTS: Approximately one-third of clients who were in full remission relapsed in the first year, and two-thirds relapsed over the full follow-up period. Predictors of relapse included male sex, less than a high school education, living independently, and lack of continued substance abuse treatment. CONCLUSIONS: After attaining full remission, clients with severe mental disorders continue to be at risk of substance abuse relapse for many years. Relapse prevention efforts should concentrate on helping clients to continue with substance abuse treatment as well as on developing housing programs that promote recovery. PMID- 16215197 TI - A focus group analysis of relapse prevention strategies for persons with substance use and mental disorders. AB - OBJECTIVE: The authors conducted a qualitative, thematic analysis of focus group data to determine the strategies and supports persons with dual diagnoses rely on in their relapse prevention efforts. METHODS: Data from four focus group sessions conducted at a large psychosocial rehabilitation center were analyzed for recurrent responses about what was most helpful in maintaining remission and grouped into major categories and subcategories. Each focus group comprised four to nine consumers who had been in remission from substance use for at least six months. A total of 27 consumers participated in the focus groups. RESULTS: The data indicate that maintaining stable housing, relying on "positive" social support, engaging in prayer or relying on a "higher power," participating in a meaningful activity, and thinking differently about life are important strategies for consumers in their attempts to stay clean. Just as frequently mentioned in the groups were conscious attempts to eat regularly, get sufficient sleep, and look presentable. CONCLUSION: Although this study was exploratory in nature, it identified areas for further qualitative study of strategies for relapse prevention among persons with dual diagnoses. PMID- 16215198 TI - Qualitative interviews on substance abuse relapse and prevention among female trauma survivors. AB - OBJECTIVE: Complex relationships among trauma, substance abuse, and mental disorders raise significant questions for the study of long-term recovery. The purpose of this qualitative study was to examine key themes in sustaining recovery among women with co-occurring disorders who had survived trauma. METHODS: In semistructured interviews conducted at one of the nine sites of the Women, Co-occurring Disorders, and Violence Study, 27 female trauma survivors described the influences they considered most important in sustaining and hindering their recovery, with an emphasis on recovery from substance abuse. Recurring themes in the interviews were identified. RESULTS: Seven themes emerged from this analysis. Four of these themes supported recovery: connection, self awareness, a sense of purpose and meaning, and spirituality. Three others served as obstacles to recovery: battles with depression and despair, destructive habits and patterns, and lack of personal control. The women in this study reported that, although caring relationships provided important supports for sustained recovery, some of these same relationships increased emotional stress and conflict and thus may impede recovery. CONCLUSIONS: It is important for women and clinicians to place a high priority on the development of boundary management and other relationship skills. In addition, clinicians need to attend to negative feelings such as boredom and loneliness and to help women develop a range of meaningful activities that are consistent with a strong sense of identity. Individual relapse prevention skills by themselves seem insufficient to sustained abstinence. PMID- 16215199 TI - Future directions in preventing relapse to substance abuse among clients with severe mental illnesses. AB - The authors review the literature on substance use disorders among persons with severe mental illnesses, including the other papers in this special section on relapse prevention, and suggest future directions. Although prevention of relapse to substance abuse has a well-developed theoretical and empirical base, this perspective has rarely been applied to persons with co-occurring severe mental illness. Research indicates that clients with co-occurring disorders are highly prone to relapse to substance abuse, even after they have attained full remission. Their risk factors include exacerbations of mental illness, social pressures within drug-using networks, lack of meaningful activities and social supports for recovery, independent housing in high-risk neighborhoods, and lack of substance abuse or dual diagnosis treatments. The evidence in hand suggests several steps: developing healthy and protective environments that are experienced as nurturing of recovery; helping people make fundamental changes in their lives, such as finding satisfying jobs, abstinent friends, networks of people who are in the process of recovery, and a sense of meaning; providing specific and individualized treatments for mental illnesses, substance use disorders, and other co-occurring problems; and developing longitudinal research on understanding and preventing relapse that addresses social context as well as biological vulnerabilities and cognitive strategies. PMID- 16215200 TI - 2005 APA Gold Award: Providing housing first and recovery services for homeless adults with severe mental illness. PMID- 16215201 TI - 2005 APA Gold Award: Improving treatment engagement and integrated care of veterans. PMID- 16215202 TI - 2005 APA Silver Awards: Silver Achievement Awards. The Child and Adolescent Services Program of the World Trade Center Healing Services, Saint Vincent Catholic Medical Centers, New York--providing trauma-related treatment to students after the terrorist attacks of September 11, 2001, and other traumatic events. PMID- 16215203 TI - Use of pulse oximetry during restraint episodes. PMID- 16215205 TI - Service use by Asian Americans. PMID- 16215206 TI - Sample for validation of jail mental health screen. PMID- 16215207 TI - Client involvement in services research. PMID- 16215208 TI - An innovative inpatient psychotherapy unit in South Africa. PMID- 16215210 TI - Endothelial survival factors and spatial completion, but not pericyte coverage of retinal capillaries determine vessel plasticity. AB - Pericyte loss and capillary regression are characteristic for incipient diabetic retinopathy. Pericyte recruitment is involved in vessel maturation, and ligand receptor systems contributing to pericyte recruitment are survival factors for endothelial cells in pericyte-free in vitro systems. We studied pericyte recruitment in relation to the susceptibility toward hyperoxia-induced vascular remodeling using the pericyte reporter X-LacZ mouse and the mouse model of retinopathy of prematurity (ROP). Pericytes were found in close proximity to vessels, both during formation of the superficial and the deep capillary layers. When exposure of mice to the ROP was delayed by 24 h, i.e., after the deep retinal layer had formed [at postnatal (p) day 8], preretinal neovascularizations were substantially diminished at p18. Mice with a delayed ROP exposure had 50% reduced avascular zones. Formation of the deep capillary layers at p8 was associated with a combined up-regulation of angiopoietin-1 and PDGF-B, while VEGF was almost unchanged during the transition from a susceptible to a resistant capillary network. Inhibition of Tie-2 function either by soluble Tie-2 or by a sulindac analog, an inhibitor of Tie-2 phosphorylation, resensitized retinal vessels to neovascularizations due to a reduction of the deep capillary network. Inhibition of Tie-2 function had no effect on pericyte recruitment. Our data indicate that the final maturation of the retinal vasculature and its resistance to regressive signals such as hyperoxia depend on the completion of the multilayer structure, in particular the deep capillary layers, and are independent of the coverage by pericytes. PMID- 16215211 TI - The modulus of elasticity of fibrillin-containing elastic fibres in the mesoglea of the hydromedusa Polyorchis penicillatus. AB - Hydromedusan jellyfish swim by rhythmic pulsation of their mesogleal bells. A single swimming muscle contracts to create thrust by ejecting water from the subumbrellar cavity. At the end of the contraction, energy stored in the deformation of the mesogleal bell powers the refilling stage, during which water is sucked back into the subumbrellar cavity. The mesoglea is a mucopolysaccharide gel reinforced with radially oriented fibres made primarily of a protein homologous to mammalian fibrillin. Most of the energy required to power the refill stroke is thought to be stored by stretching these fibres. The elastic modulus of similar fibrillin-rich fibres has been measured in other systems and found to be in the range of 0.2 to 1.1 MPa. In this paper, we measured the diameters of the fibres, their density throughout the bell, and the mechanical behaviour of the mesoglea, both in isolated samples and in an intact bell preparation. Using this information, we calculated the stiffness of the fibres of the hydromedusa Polyorchis penicillatus, which we found to be approximately 0.9 MPa, similar in magnitude to other species. This value is two orders of magnitude more compliant than the stiffness of the component fibrillin microfibrils previously reported. We show that the structure of the radial fibres can be modelled as a parallel fibre-reinforced composite and reconcile the stiffness difference by reinterpreting the previously reported data. We separate the contributions to the bell elasticity of the fibres and mesogleal matrix and calculate the energy storage capacity of the fibres using the calculated value of their stiffness and measured densities and diameters. We conclude that there is enough energy potential in the fibres alone to account for the energy required to refill the subumbrellar cavity. PMID- 16215212 TI - Submerged swimming of the great cormorant Phalacrocorax carbo sinensis is a variant of the burst-and-glide gait. AB - Cormorants are water birds that forage by submerged swimming in search and pursuit of fish. Underwater they swim by paddling with both feet simultaneously in a gait that includes long glides between consecutive strokes. At shallow swimming depths the birds are highly buoyant as a consequence of their aerial lifestyle. To counter this buoyancy cormorants swim underwater with their body at an angle to the swimming direction. This mechanical solution for foraging at shallow depth is expected to increase the cost of swimming by increasing the drag of the birds. We used kinematic analysis of video sequences of cormorants swimming underwater at shallow depth in a controlled research setup to analyze the swimming gait and estimate the resultant drag of the birds during the entire paddling cycle. The gliding drag of the birds was estimated from swimming speed deceleration during the glide stage while the drag during active paddling was estimated using a mathematical ;burst-and-glide' model. The model was originally developed to estimate the energetic saving from combining glides with burst swimming and we used this fact to test whether the paddling gait of cormorants has similar advantages. We found that swimming speed was correlated with paddling frequency (r=0.56, P<0.001, N=95) where the increase in paddling frequency was achieved mainly by shortening the glide stage (r=-0.86, P<0.001, N=95). The drag coefficient of the birds during paddling was higher on average by two- to threefold than during gliding. However, the magnitude of the drag coefficient during the glide was positively correlated with the tilt of the body (r=0.5, P<0.003, N=35) and negatively correlated with swimming speed (r=-0.65, P<0.001, N=35), while the drag coefficient during the stroke was not correlated with tilt of the body (r=-0.11, P>0.5, N=35) and was positively correlated with swimming speed (r=0.41, P<0.015, N=35). Therefore, the difference between the drag coefficient during the glide and during propulsion diminished at lower speeds and larger tilt. The mean drag of the birds for a single paddling cycle at an average swimming speed of 1.5 m s(-1) was 5.5+/-0.68 N. The burst-and-glide model predicts that energy saving from using burst-and-glide in the paddling cycle is limited to relatively fast swimming speeds (>1.5 m s(-1)), but that as the birds dive deeper (>1 m where buoyancy is reduced), the burst-and-glide gait may become beneficial even at lower speeds. PMID- 16215213 TI - Chloride turnover and ion-transporting activities of yolk-sac preparations (yolk balls) separated from Mozambique tilapia embryos and incubated in freshwater and seawater. AB - We have recently established a unique in vitro experimental model for mitochondrion-rich cell (MRC) research, a ;yolk-ball' incubation system, in which the yolk sac is separated from the embryonic body of Mozambique tilapia embryos and subjected to in vitro incubation. To evaluate the ion-transporting property of the yolk balls, we examined Cl- content and turnover in yolk balls incubated in freshwater and seawater for 48 h, and distribution patterns of three ion transporters, Na+/K+-ATPase, Na+/K+/2Cl- cotransporter (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR), in MRCs in the yolk-sac membrane. The Cl- turnover rate measured by whole-body influx of 36Cl- was about 60 times higher in yolk balls in seawater than in freshwater, while there was no essential difference in Cl- content between them. Na+/K+-ATPase-immunoreactive MRCs were larger in yolk balls from seawater than yolk balls from freshwater. Distribution patterns of ion-transporting proteins allowed us to classify MRCs in freshwater yolk balls into three types: cells showing only basolateral Na+/K+-ATPase, cells showing basolateral Na+/K+-ATPase and apical NKCC, and cells showing basolateral Na+/K+-ATPase and basolateral NKCC. The seawater yolk balls, on the other hand, were characterized by the appearance of MRCs possessing basolateral Na+/K+ ATPase, basolateral NKCC and apical CFTR. Those seawater-type MRCs were considered to secrete Cl- through the CFTR-positive apical opening to cope with diffusional Cl- influx. These findings indicate that the yolk balls preserve the Cl- transporting property of intact embryos, ensuring the propriety of the yolk ball as an in vitro experimental model for the yolk-sac membrane that contains MRCs. PMID- 16215214 TI - Differential freshwater adaptation in juvenile sea-bass Dicentrarchus labrax: involvement of gills and urinary system. AB - The effects of long-term freshwater acclimatization were investigated in juvenile sea-bass Dicentrarchus labrax to determine whether all sea-bass juveniles are able to live in freshwater and to investigate the physiological basis of a successful adaptation to freshwater. This study particularly focused on the ability of sea-bass to maintain their hydromineral balance in freshwater and on their ion (re)absorbing abilities through the gills and kidneys. Two different responses were recorded after a long-term freshwater acclimatization. (1) Successfully adapted sea-bass displayed standard behavior; their blood osmolality was maintained almost constant after the freshwater challenge, attesting to their efficient hyperosmoregulation. Their branchial and renal Na+/K+-ATPase abundance and activity were high compared to seawater fish due to a high number of branchial ionocytes and to the involvement of the urinary system in active ion reabsorption, producing hypotonic urine. (2) Sea-bass that had not successfully adapted to freshwater were recognized by abnormal schooling behavior. Their blood osmolality was low (30% lower than in the successfully adapted sea-bass), which is a sign of acute osmoregulatory failure. High branchial Na+/K+-ATPase abundance and activity compared to successfully adapted fish were coupled to a proliferation of gill chloride cells, whose ultrastructure did not display pathological signs. The large surface used by the gill chloride cells might negatively interfere with respiratory gas exchanges. In their urinary system, enzyme abundance and activity were low, in accordance with the observed lower density of the kidney tubules. Urine was isotonic to blood in unsuccessfully adapted fish, ruling out any participation of the kidney in hyperosmoregulation. The kidney failure seems to generate a compensatory ion absorption through increased gill activity, but net ion loss through urine seems higher than ion absorption by the gills, leading to lower hyper-osmoregulatory performance and to death. PMID- 16215215 TI - Bioenergetics and diving activity of internesting leatherback turtles Dermochelys coriacea at Parque Nacional Marino Las Baulas, Costa Rica. AB - Physiology, environment and life history demands interact to influence marine turtle bioenergetics and activity. However, metabolism and diving behavior of free-swimming marine turtles have not been measured simultaneously. Using doubly labeled water, we obtained the first field metabolic rates (FMRs; 0.20-0.74 W kg( 1)) and water fluxes (16-30% TBW day(-1), where TBW=total body water) for free ranging marine turtles and combined these data with dive information from electronic archival tags to investigate the bioenergetics and diving activity of reproductive adult female leatherback turtles Dermochelys coriacea. Mean dive durations (7.8+/-2.4 min (+/-1 s.d.), bottom times (2.7+/-0.8 min), and percentage of time spent in water temperatures (Tw) < or =24 degrees C (9.5+/ 5.7%) increased with increasing mean maximum dive depths (22.6+/-7.1 m; all P< or =0.001). The FMRs increased with longer mean dive durations, bottom times and surface intervals and increased time spent in Tw< or =24 degrees C (all r2> or =0.99). This suggests that low FMRs and activity levels, combined with shuttling between different water temperatures, could allow leatherbacks to avoid overheating while in warm tropical waters. Additionally, internesting leatherback dive durations were consistently shorter than aerobic dive limits calculated from our FMRs (11.7-44.3 min). Our results indicate that internesting female leatherbacks maintained low FMRs and activity levels, thereby spending relatively little energy while active at sea. Future studies should incorporate data on metabolic rate, dive patterns, water temperatures, and body temperatures to develop further the relationship between physiological and life history demands and marine turtle bioenergetics and activity. PMID- 16215216 TI - Encoding spatial information in the waggle dance. AB - Apis mellifera bees execute waggle dances to recruit other bees to desirable food sources. Several components of the waggle dance are correlated with the direction of and the distance to food. Moreover, recruits use the spatial information encoded in the dance to locate the signalled food. However, although dance communication has been studied extensively, little is known about how the dancers combine the compass (direction) and the odometric (distance) information they acquire during the foraging flight. In the present study, we analysed the encoding of spatial information in the waggle dance by manipulating the navigational information provided to dancing bees. To this end, we took advantage of the bees' visually driven odometer. We found that the waggle dance basically encodes information on the distance gauged during the outbound (hive-to-food) flight. However, it does not necessarily refer to a global vector based on path integration of the outbound flight. Whenever the direction connecting the subjective food location and the hive does not match the direction of the global vector, dancers refer to a direction close to that of the shortcut connecting the actual food location and the hive. Moreover, in our experiments, this direction was close to that of the inbound (food-to-hive) flight, indicating that landmark based information is computed during the inbound flight and that it may strongly affect the encoding of directional information in the waggle dance. Moreover, we found that the bees' experience of the terrain modulates the encoding of spatial information in the waggle dance, suggesting that interactions between path integration and visual landmarks are computed in the context of dance communication. PMID- 16215217 TI - Visual control of flight speed in honeybees. AB - Visual control of flight speed in honeybees (Apis mellifera L.) was investigated by training them to fly through a specially constructed tunnel in which the motion, contrast and texture of the patterns lining the walls could be varied. Manipulation of pattern motion revealed that the speed of flight is controlled by regulating the image motion that is experienced by the eyes. Flight speed is surprisingly robust to changes in the contrast and/or spatial texture of the visual environment, suggesting that the underlying movement-detecting mechanisms estimate the speed of image motion in the eye largely independently of these parameters. This ensures that flight speed depends primarily on the distances to nearby surfaces and not upon their particular visual properties, such as contrast or visual texture. The removal of image motion cues drastically compromises the regulation of flight speed, underscoring their role in this function. PMID- 16215218 TI - Mechanical and morphological properties of different muscle-tendon units in the lower extremity and running mechanics: effect of aging and physical activity. AB - The objectives of this work were (i) to investigate whether chronic endurance running is a sufficient stimulus to counteract the age-related changes in the mechanical and morphological properties of human triceps surae (TS) and quadriceps femoris (QF) muscle-tendon units (MTUs) by comparing runners and non active subjects at different ages (young and old), (ii) to identify adaptational phenomena in running mechanics due to age-related changes in the mechanical and morphological properties of the TS and QF MTUs, and finally (iii) to examine whether chronic endurance-running exercise is associated with adaptational effects on running characteristics in old and young adults. The investigation was conducted on 30 old and 19 young adult males divided into two subgroups according to their running activity: endurance-runners vs non-active. To analyse the properties of the MTUs, all subjects performed isometric maximal voluntary (MVC) ankle plantarflexion and knee extension contractions at 11 different MTU lengths on a dynamometer. The activation of the TS and QF during MVC was estimated by surface electromyography. The gastrocnemius medialis and the vastus lateralis and their distal aponeuroses were visualized by ultrasonography at rest and during MVC, respectively. Ground reaction forces and kinematic data were recorded during running trials at 2.7 m s(-1). The TS and QF MTU capacities were reduced with aging (lower muscle strength and lower tendon stiffness). Runners and non-active subjects had similar MTU properties, suggesting that chronic endurance-running exercise does not counteract the age-related degeneration of the MTUs. Runners showed a higher mechanical advantage for the QF MTU while running (lower gear ratio) compared to non-active subjects, indicating a task-specific adaptation even at old age. Older adults reacted to the reduced capacities of their MTUs by increasing running safety (higher duty factor, lower flight time) and benefitting from a mechanical advantage for the TS MTU, lower rate of force generation and force generation per meter distance. We suggest that the improvement in running mechanics in the older adults happens due to a perceptual motor recalibration and a feed-forward adaptation of the motor task aimed at decreasing the disparity between the reduced capacity of the MTUs and the running effort. PMID- 16215219 TI - The role of UV in crab spider signals: effects on perception by prey and predators. AB - Australian crab spiders Thomisus spectabilis sit on the petals of flowers and ambush prey such as honeybees. White-coloured T. spectabilis reflect in the UV (UV+ spiders) and previous research has shown that their presence, curiously, attracts honeybees to daisies. We applied an UV-absorber (Parsol) to create UV absorbing (UV-) spiders that did not reflect any light below 395 nm wavelength. These physical changes of visual signals generated by crab spiders caused honeybees to avoid flowers with UV- spiders on their petals. They also affected the perception of UV- spiders by honeybees and a potential avian predator (blue tits). Compared to UV+ spiders, UV- spiders produced less excitation of the UV photoreceptors in honeybees and blue tits, which translated into a reduced UV receptor contrast and a reduced overall colour contrast between UV- spiders and daisy petals. Our results reveal that a clean physical elimination of reflection in the UV range affects perception in predators and prey and ultimately changes the behaviour of prey. PMID- 16215220 TI - Effect of food quality, distance and height on thoracic temperature in the stingless bee Melipona panamica. AB - Stingless bees (Apidae, Meliponini) can recruit nestmates to good food sources. We present the first data showing that recruiting meliponine foragers at feeders and inside nests regulate their thoracic temperature according to net food profitability. Using infrared thermography, we found that Melipona panamica foragers elevated their thoracic temperature at profitable food sources (higher sucrose concentration, closer to the nest). At food sources, there is an increase of approximately 0.9 degrees C in thoracic temperature (Tth) per 1 mol l(-1) increase in sucrose concentration (at 20 m distance from nest: 1 mol l(-1) sucrose concentration, Tth=36.6+/-0.8 degrees C, T(a)=31.3+/-0.5 degrees C; 2.5 mol l(-1) sucrose concentration, Tth=36.9+/-0.6 degrees C, Ta=29.9+/-0.2 degrees C). Inside the nest, the difference between thoracic temperature Tth and ambient air temperature Ta (DeltaTnest) decreases by 0.4 degrees C with each 100 m increase in feeder-to-nest distance and increases by 0.1 degrees C per 1 mol l( 1) increase in sucrose concentration. The Tth of returning foragers was significantly higher at all tested sucrose concentrations (1.0-2.5 mol l(-1) sucrose concentration) and distances (25-437 m) as compared to Ta (at 2.5 mol l( 1) sucrose concentration: 25 m distance from nest, intranidal Tth=30.2+/-1.3 degrees C, Ta=24.8+/-0.7 degrees C; 437 m distance from nest, intranidal Tth=28.6+/-1.7 degrees C, Tnest=25.4+/-1.4 degrees C). For highly profitable food sources (2.5 mol l(-1) sucrose concentration and < or =100 m from the nest), forager Tth was slightly higher than that of randomly chosen control bees inside the nest. PMID- 16215221 TI - Look and turn: landmark-based goal navigation in honey bees. AB - This report describes the piloting mechanisms employed by honey bees during their final approach to a goal. Conceptually applying a bottom-up approach, we systematically varied the position, number and appearance landmarks associated with a rewarded target location within a large, homogenous flight tent. The flight behavior measured under various conditions is well explained with visuo motor control loops that link perceived landmarks with appropriate turning responses. This view is consistent with the requirement of prolonged reinforcement learning for efficient goal navigation. A simple model is able to provide a comprehensive explanation for diverse flight patterns that range from convoluted searching behavior to highly idiosyncratic approaches, depending on the experimental context. Our results challenge the prevalent notion that honey bees employ image matching for visual guidance toward a goal site. Basic visuo motor control loops may better meet the high demands for robust and fast flight control, which could serve as a powerful bio-mimetic design principle for micro robotic aircraft. PMID- 16215222 TI - Electrical properties and fusion dynamics of in vitro membrane vesicles derived from separate parts of the contractile vacuole complex of Paramecium multimicronucleatum. AB - The contractile vacuole complex of Paramecium multimicronucleatum transforms into membrane-bound vesicles on excision from the cell. The I-V relationship was linear in a voltage range of -80 to +80 mV in all vesicles, despite being derived from different parts of the contractile vacuole complex. No voltage-gated unit currents were observed in membrane patches from the vesicles. Vesicles derived from the radial arm showed a membrane potential of >10 mV, positive with reference to the cytosol, while those derived from the contractile vacuole showed a residual (<5 mV) membrane potential. The electrogenic V-ATPases in the decorated spongiome are responsible for the positive potential, and Cl- leakage channels are responsible for the residual potential. The specific resistance of the vesicle membrane (approximately 6 kOmega cm2) increased, while the membrane potential shifted in a negative direction when the vesicle rounded. An increase in the membrane tension (to approximately 5 x 10(-3) N m(-1)) is assumed to reduce the Cl- leakage conductance. It is concluded that neither voltage- nor mechano-sensitive ion channels are involved in the control of the fluid segregation and membrane dynamics that govern fluid discharge cycles in the contractile vacuole complex. The membrane vesicles shrank when the external osmolarity was increased, and swelled when the osmolarity was decreased, implying that the contractile vacuole complex membrane is water permeable. The water permeability of the membrane was 4-20 x 10(-7) microm s(-1) Pa(-1). The vesicles containing radial arm membrane swelled after initially shrinking when exposed to higher external osmolarity, implying that the V-ATPases energize osmolyte transport mechanisms that remain functional in the vesicle membrane. The vesicles showed an abrupt (<30 ms), slight, slackening after rounding to the maximum extent. Similar slackening was also observed in the contractile vacuoles in situ before the opening of the contractile vacuole pore. A slight membrane slackening seems to be an indispensable requirement for the contractile vacuole membrane to fuse with the plasma membrane at the pore. The contractile vacuole complex derived membrane vesicle is a useful tool for understanding not only the biological significance of the contractile vacuole complex but also the molecular mechanisms of V-ATPase activity. PMID- 16215223 TI - An updated catalogue of salivary gland transcripts in the adult female mosquito, Anopheles gambiae. AB - Salivary glands of blood-sucking arthropods contain a variety of compounds that prevent platelet and clotting functions and modify inflammatory and immunological reactions in the vertebrate host. In mosquitoes, only the adult female takes blood meals, while both sexes take sugar meals. With the recent description of the Anopheles gambiae genome, and with a set of approximately 3000 expressed sequence tags from a salivary gland cDNA library from adult female mosquitoes, we attempted a comprehensive description of the salivary transcriptome of this most important vector of malaria transmission. In addition to many transcripts associated with housekeeping functions, we found an active transposable element, a set of Wolbachia-like proteins, several transcription factors, including Forkhead, Hairy and doublesex, extracellular matrix components and 71 genes coding for putative secreted proteins. Fourteen of these 71 proteins had matching Edman degradation sequences obtained from SDS-PAGE experiments. Overall, 33 transcripts are reported for the first time as coding for salivary proteins. The tissue and sex specificity of these protein-coding transcripts were analyzed by RT-PCR and microarray experiments for insight into their possible function. Notably, two gene products appeared to be differentially spliced in the adult female salivary glands, whereas 13 contigs matched predicted intronic regions and may include additional alternatively spliced transcripts. Most An. gambiae salivary proteins represent novel protein families of unknown function, potentially coding for pharmacologically or microbiologically active substances. Supplemental data to this work can be found at http://www.ncbi.nlm.nih.gov/projects/omes/index.html#Ag2. PMID- 16215224 TI - Functional and structural optimization of the respiratory system of the bat Tadarida brasiliensis (Chiroptera, Molossidae): does airway geometry matter? AB - We studied structure and function of the respiratory system in the bat Tadarida brasiliensis and compared it with those of two species of rodents, Abrothrix andinus and A. olivaceus. Tadarida brasiliensis had lower resting oxygen consumption, but higher maximum oxygen consumption and aerobic scope, than the rodents. The blood-gas barrier of the bat was thinner and its relative lung size was larger; however, alveolar surface density was similar among the three species. In consequence, T. brasiliensis has an oxygen diffusion capacity two or three times higher than that of the rodents. In Tadarida brasiliensis the characteristics of the lung were accompanied by geometrical changes in the proximal airway, such as high physical optimization as a consequence of small variations in the symmetry and the scaling ratio of the bronchial diameters. These may constitute an efficient way to save energy in respiratory mechanics and are the first report of airway adjustments to decrease entropy generation in bats. PMID- 16215225 TI - Isolation, characterization, and pericycle-specific transcriptome analyses of the novel maize lateral and seminal root initiation mutant rum1. AB - The monogenic recessive maize (Zea mays) mutant rootless with undetectable meristems 1 (rum1) is deficient in the initiation of the embryonic seminal roots and the postembryonic lateral roots at the primary root. Lateral root initiation at the shoot-borne roots and development of the aerial parts of the mutant rum1 are not affected. The mutant rum1 displays severely reduced auxin transport in the primary root and a delayed gravitropic response. Exogenously applied auxin does not induce lateral roots in the primary root of rum1. Lateral roots are initiated in a specific cell type, the pericycle. Cell-type-specific transcriptome profiling of the primary root pericycle 64 h after germination, thus before lateral root initiation, via a combination of laser capture microdissection and subsequent microarray analyses of 12k maize microarray chips revealed 90 genes preferentially expressed in the wild-type pericycle and 73 genes preferentially expressed in the rum1 pericycle (fold change >2; P-value <0.01; estimated false discovery rate of 13.8%). Among the 51 annotated genes predominately expressed in the wild-type pericycle, 19 genes are involved in signal transduction, transcription, and the cell cycle. This analysis defines an array of genes that is active before lateral root initiation and will contribute to the identification of checkpoints involved in lateral root formation downstream of rum1. PMID- 16215227 TI - Functions of early (AP-2) and late (AIP1/ALIX) endocytic proteins in equine infectious anemia virus budding. AB - The proline-rich L domains of human immunodeficiency virus 1 (HIV-1) and other retroviruses interact with late endocytic proteins during virion assembly and budding. In contrast, the YPDL L domain of equine infectious anemia virus (EIAV) is apparently unique in its reported ability to interact both with the mu2 subunit of the AP-2 adaptor protein complex and with ALG-2-interacting protein 1 (AIP1/Alix) protein factors involved in early and late endosome formation, respectively. To define further the mechanisms by which EIAV adapts vesicle trafficking machinery to facilitate virion production, we have examined the specificity of EIAV p9 binding to endocytic factors and the effects on virion production of alterations in early and late endocytic protein expression. The results of these studies demonstrated that (i) an approximately 300-residue region of AIP1/Alix-(409-715) was sufficient for binding to the EIAV YPDL motif; (ii) overexpression of AIP1/Alix or AP-2 mu2 subunit specifically inhibited YPDL mediated EIAV budding; (iii) virion budding from a replication-competent EIAV variant with its L domain replaced by the HIV PTAP sequence was inhibited by wild type or mutant mu2 to a level similar to that observed when a dominant-negative mutant of Tsg101 was expressed; and (iv) overexpression or siRNA silencing of AIP1/Alix and AP-2 revealed additive suppression of YPDL-mediated EIAV budding. Taken together, these results indicated that both early and late endocytic proteins facilitate EIAV production mediated by either YPDL or PTAP L domains, suggesting a comprehensive involvement of endocytic factors in retroviral assembly and budding that can be accessed by distinct L domain specificities. PMID- 16215228 TI - The thylakoid delta pH/delta psi are not required for the initial stages of Tat dependent protein transport in tobacco protoplasts. AB - The twin-arginine translocation (Tat) system transports folded proteins across the chloroplast thylakoid membrane and bacterial plasma membrane. In vitro import assays have pointed to a key role for the thylakoid delta pH in the initial assembly of the full translocon from two subcomplexes; more generally, the delta pH is believed to provide the overall driving force for translocation. Here, we have studied the role of the delta pH in vivo by analyzing the translocation of Tat substrates in transfected tobacco protoplasts. We show that the complete maturation of the precursor of the 23-kDa lumenal protein (pre-23K) and of a fusion of the 23K presequence linked to green fluorescent protein (pre-GFP) are unaffected by dissipation of the delta pH. High level expression of Tat substrates in protoplasts has recently been shown to result in "translocation reversal" in that a large proportion of a given substrate is partially translocated across the thylakoid membrane, processed to the mature size, and returned to the stroma. However, the efficiency of translocation of pre-23K is undiminished in the absence of the delta pH and/or delta psi, and the rate and extent of maturation of both pre-23K and pre-GFP by the lumen-facing processing peptidase is similarly unaffected. These data demonstrate that the proton motive force is not required for the functional assembly of the Tat translocon and the initial stages of translocation in higher plant chloroplasts in vivo. We conclude that unknown factors play an influential role in both the mechanism and energetics of this system under in vivo conditions. PMID- 16215229 TI - The solvent protection of alzheimer amyloid-beta-(1-42) fibrils as determined by solution NMR spectroscopy. AB - Alzheimer disease is a neurodegenerative disorder that is tightly linked to the self-assembly and amyloid formation of the 39-43-residue-long amyloid-beta (Abeta) peptide. Considerable evidence suggests a correlation between Alzheimer disease development and the longer variants of the peptide, Abeta-(1-42/43). Currently, a molecular understanding for this behavior is lacking. In the present study, we have investigated the hydrogen/deuterium exchange of Abeta-(1-42) fibrils under physiological conditions, using solution NMR spectroscopy. The obtained residue-specific and quantitative map of the solvent protection within the Abeta-(1-42) fibril shows that there are two protected core regions, Glu11 Gly25 and Lys28-Ala42, and that the residues in between, Ser26 and Asn27, as well as those in the N terminus, Asp1-Tyr10, are solvent-accessible. This result reveals considerable discrepancies when compared with a previous investigation on Abeta-(1-40) fibrils and suggests that the additional residues in Abeta-(1-42), Ile41 and Ala42, significantly increase the solvent protection and stability of the C-terminal region Lys28-Ala42. Consequently, our findings provide a molecular explanation for the increased amyloidogenicity and toxicity of Abeta-(1-42) compared with shorter Abeta variants found in vivo. PMID- 16215230 TI - Expansion of substrate specificity of cytochrome P450 2A6 by random and site directed mutagenesis. AB - The natural product indole is a substrate for cytochrome P450 2A6. Mutagenesis of P450 2A6 was done to expand its capability in the oxidization of bulky substituted indole compounds, which are not substrates for the wild-type enzyme or the double mutant L240C/N297Q, as determined in our previous work (Wu, Z.-L., Aryal, P., Lozach, O., Meijer, L., and Guengerich, F. P. (2005) Chem. Biodivers. 2, 51-65). Error-prone PCR and site-directed mutagenesis led to the identification of two critical amino acid residue changes (N297Q and I300V) that achieve the purpose. The new mutant (N297Q/I300V) was able to oxidize both 4- and 5-benzyloxy(OBzl)indoles to form colored products. Both changes were required for oxidation of these bulky substrates. The colored product derived from 5-OBzl indole was mainly 5,5'-di-OBzl-indirubin, whereas the dominant blue dye isolated upon incubations with 4-OBzl-indole was neither an indigo nor an indirubin. Two dimensional NMR experiments led to assignment of the structure as 4-OBzl-2-(4' OBzl-1',7'-dihydro-7'-oxo-6'H-indol-6'-ylidene)indolin-3-one, in which a pyrrole ring and a benzene ring are connected with a double bond instead of the pyrrole pyrrole connection of other indigoids. Monomeric oxidation products were also isolated and characterized; three phenols (4-OBzl-1H-indol-5-ol, 4-OBzl-1H-indol 6-ol, and 4-OBzl-1H-indol-7-ol) and one quinone (4-OBzl-1H-indole-6,7-dione, the postulated immediate precursor of the final blue dye) were identified. The results are interpreted in the context of a crystal structure of a P450 2A6 coumarin complex. The I300V change opens an additional pocket to accommodate the OBzl bulk. The N2297Q change is postulated to generate a hydrogen bond between Gln and the substrate oxygen. Thus, the substrate specificity of P450 2A6 was expanded, and new products were obtained in this study. PMID- 16215231 TI - Extended upstream A-T sequence increases T7 promoter strength. AB - Bacteriophage T7 promoters contain a consensus sequence from -17 to +6 relative to the transcription start site, +1. In addition, the strong class III promoters are characterized by an extended AT-rich region upstream of -17, which is often interrupted by one or more GC base pairs in the weaker class II promoters. Herein we studied the role of the AT-rich region upstream of -17 in transcription regulation of T7 RNA polymerase. Equilibrium DNA binding studies with promoter fragments of consensus sequence truncated at various positions between -17 and 27 showed that the polymerase-promoter complex is significantly stabilized as the upstream AT-rich sequence is extended to and beyond -22. Similarly, promoters in which the AT-rich region from -17 to -22 is interrupted by several GC base pairs showed weak binding. Kinetic studies indicated that the presence of extended AT rich sequence slows the dissociation rate constant of the polymerase-promoter complex and slightly stimulates the association rate constant, thereby increasing the stability of the complex. Measurement of the transcription activity revealed that the extended AT-rich region does not affect the kinetics of abortive synthesis up to the formation of 8-nucleotide RNA but causes accumulation of longer abortive products between 9 and 13 nucleotides. The observed effects of the upstream DNA region were AT sequence-specific, and the results suggested a larger role for the extended AT-rich sequence that has been unappreciated previously. We propose that the AT-rich DNA sequence upstream of -17 plays a role in modulating the efficiency of transcription initiation by affecting both the affinity of T7 RNA polymerase for the promoter and the efficiency of promoter clearance. PMID- 16215232 TI - Individual phosphoinositide 3-kinase C2alpha domain activities independently regulate clathrin function. AB - Phosphoinositide 3-kinase C2alpha (PI3K-C2alpha) is a member of the class II PI-3 kinases, defined by the presence of a second C2 domain at their C termini. The cellular functions of the class II enzymes are incompletely understood, though they have been implicated in receptor activation pathways initiated by epidermal growth factor, insulin, and chemokines. PI3K-C2alpha was recently found to be localized to clathrin-coated membranes in the trans-Golgi network and at endocytic sites on the plasma membrane. Further, a specific binding site was identified for clathrin on the N terminus of PI3K-C2alpha, whose occupancy resulted in lipid kinase activation. Expression of PI3K-C2alpha in cells dramatically affected clathrin distribution and function in cells, leading to accumulation of intracellular clathrin-coated structures, which are visualized here at the ultrastructural level, and inhibition of clathrin-mediated transport from both the plasma membrane and the trans-Golgi network. In this study we have demonstrated that the isolated clathrin binding domain of PI3K-C2alpha can drive clathrin lattice assembly and that both it and the lipid kinase activity of the protein can independently modulate clathrin distribution and function when expressed in cells. Together, these results suggest that PI3K-C2alpha employs both protein-protein interaction and localized production of 3-phosphoinositides to affect clathrin dynamics at sites of membrane budding and targeting. PMID- 16215238 TI - Changes to medicines legislation: issues discussed at BVA Congress 2005. PMID- 16215239 TI - Equal opportunities? Demographic and financial pressures on the profession. PMID- 16215234 TI - The tissue factor requirement in blood coagulation. AB - Formation of thrombin is triggered when membrane-localized tissue factor (TF) is exposed to blood. In closed models of this process, thrombin formation displays an initiation phase (low rates of thrombin production cause platelet activation and fibrinogen clotting), a propagation phase (>95% of thrombin production occurs), and a termination phase (prothrombin activation ceases and free thrombin is inactivated). A current controversy centers on whether the TF stimulus requires supplementation from a circulating pool of blood TF to sustain an adequate procoagulant response. We have evaluated the requirement for TF during the progress of the blood coagulation reaction and have extended these analyses to assess the requirement for TF during resupply ("flow replacement"). Elimination of TF activity at various times during the initiation phase indicated: a period of absolute dependence (<10 s); a transitional period in which the dependence on TF is partial and decreases as the reaction proceeds (10 240 s); and a period in which the progress of the reaction is TF independent (>240 s). Resupply of reactions late during the termination phase with fresh reactants, but no TF, yielded immediate bursts of thrombin formation similar in magnitude to the original propagation phases. Our data show that independence from the initial TF stimulus is achieved by the onset of the propagation phase and that the ensemble of coagulation products and intermediates that yield this TF independence maintain their prothrombin activating potential for considerable time. These observations support the hypothesis that the transient, localized expression of TF is sufficient to sustain a TF-independent procoagulant response as long as flow persists. PMID- 16215233 TI - Substrate recognition by the human fatty-acid synthase. AB - The human fatty-acid synthase (HFAS) is a potential target for anti-tumor drug discovery. As a prelude to the design of compounds that target the enoyl reductase (ER) component of HFAS, the recognition of NADPH and exogenous substrates by the ER active site has been investigated. Previous studies demonstrate that modification of Lys-1699 by pyridoxal 5'-phosphate results in a specific decrease in ER activity. For the overall HFAS reaction, the K1699A and K1699Q mutations reduced kcat and kcat/KNADPH by 8- and 600-fold, respectively (where KNADPH indicates the Km value for NADPH). Thus, Lys-1699 contributes 4 kcal/mol to stabilization of the rate-limiting transition state following NADPH binding, while also stabilizing the most stable ground state after NADPH binding by 3 kcal/mol. A similar effect of the mutations on the ER partial reaction was observed, in agreement with the proposal that Lys-1699 is located in the ER NADPH binding site. Most unexpectedly, however, both kcat and kcat/KNADPH for the beta ketoacyl reductase (BKR) reaction were also impacted by the Lys-1699 mutations, raising the possibility that the ER and BKR activities share a single active site. However, based on previous data indicating that the two reductase activities utilize distinct cofactor binding sites, mutagenesis of Lys-1699 is hypothesized to modulate BKR activity via allosteric effects between the ER and BKR NADPH sites. PMID- 16215243 TI - Diagnostic value of cytology of bronchoalveolar fluid for lung diseases of sheep. AB - Bronchoalveolar lavage fluid was collected postmortem from the lungs of 113 sheep, and total and differential cell counts were analysed in relation to the presence of gross and microscopic lung pathology. The diffuse lung diseases, maedi and adenomatosis, were both characterised by an increase in overall cellularity and by increases in the percentages of lymphocytes and neutrophils, respectively. Focal parasitic lung disease was characterised by an increase in the percentage of eosinophils and mast cells. Consolidated lung lesions were characterised by a slight increase in cellularity but no change in the differential cell profile. In regions of parasitised and consolidated lungs without lesions the differential cell profile was consistent with focal lung pathology, although the slight increase in cellularity observed in the consolidated regions was not observed in the regions without lesions. A decision tree was developed to facilitate the interpretation and indicate the likely predictive capacity of the differential cytology of the fluid. PMID- 16215244 TI - Morbidity of Swedish horses insured for veterinary care between 1997 and 2000: variations with age, sex, breed and location. AB - The aim of this study was to evaluate the potential usefulness of the database maintained by the Swedish insurance company Agria for providing disease statistics on Swedish horses. The demography of the horses insured for veterinary care during the period 1997 to 2000 was recorded and the incidence of morbidity, defined as horses that required veterinary care that cost more than the policy excess, was calculated. Yearly incidences were calculated for horses that required veterinary care at least once, first overall, and then for horses with complete insurance, by sex, age, breed group, breed, location and human population density. Poisson regression was applied to a multivariable model to produce estimates of relative risk adjusted for other factors in the model, such as age. The total number of horse-years at risk for those with complete insurance was over 72,000 each year. The annual incidence rate for horses that required veterinary care at least once varied from 1080 to 1190 events per 10,000 horse years at risk; for geldings the averaged incidence rate was 1398 events, for mares it was 1042 events, and for stallions it was 780 events per 10,000 horse years at risk. There were considerable variations in incidence rate between breeds. PMID- 16215245 TI - Effects of meloxicam on the haemostatic profile of dogs undergoing orthopaedic surgery. AB - The buccal mucosal bleeding time (BMBT), prothrombin time (PT), activated partial thromboplastin time (APTT) and intraoperative bleeding score (IBS) of 38 dogs that underwent orthopaedic surgical procedures and received meloxicam orally and/or parenterally were measured. Fourteen of the dogs (group A) received a single subcutaneous dose of 0.2 mg/kg meloxicam at premedication, 18 dogs (group B) received 0.1 mg/kg meloxicam orally daily for five days followed by a single subcutaneous dose of 0.2 mg/kg meloxicam preoperatively, and six dogs (group C) received 0.5 ml of normal saline subcutaneously at premedication. No statistically significant differences among the groups were detected in relation to the mean (SD) values of BMBT, PT and IBS before and after the surgery, or in the values of APTT in group A. In group B there was a small but significant increase in APTT after the surgery, but all the measurements were within the normal range for dogs. PMID- 16215246 TI - Antigenic relationship of avian strains of three Bordetella species determined by ELISA. PMID- 16215247 TI - Isolation of bovine viral diarrhoea viruses from bison. PMID- 16215248 TI - Actinomyces naeslundii infection in an African hedgehog (Atelerix albiventris) with mandibular osteomyelitis and cellulitis. PMID- 16215249 TI - Haemolytic anaemia in a heifer with bovine viral diarrhoea and mucosal disease complex. PMID- 16215250 TI - Suicides among veterinary surgeons. PMID- 16215251 TI - RUMA guidelines on the responsible use of antimicrobials in farm animals. PMID- 16215252 TI - Effect of commercial diets on cats with chronic renal insufficiency. PMID- 16215253 TI - Ingluvitis and oesophagitis in wild finches. PMID- 16215254 TI - Ingluvitis and oesophagitis in wild finches. PMID- 16215256 TI - VAT on veterinary prescriptions. PMID- 16215257 TI - Epidermolysis bullosa in cattle in the UK. PMID- 16215259 TI - Reevaluation of the role of the multidrug-resistant P-glycoprotein in cellular cholesterol homeostasis. AB - The multidrug resistance P-glycoprotein (P-gp) was recently proposed to redistribute cholesterol in the plasma membrane, suggesting that P-gp could modulate cholesterol efflux to cholesterol acceptors. To address this hypothesis and to reevaluate the role of P-gp in cholesterol homeostasis, we first analyzed the role of P-gp expression on cholesterol efflux in P-gp stably transfected drug selected LLC-MDR1 cells. Cholesterol efflux to methyl-beta-cyclodextrin (CD) was 4-fold higher in LLC-MDR1 cells compared with control LLC-PK1 cells, indicating that the accessible pool of plasma membrane cholesterol was increased by P-gp expression. However, using the P-gp-inducible cells lines HeLa MDR-Tet and 77.1 MDR-Tet, cholesterol efflux to CD, apolipoprotein A-I, or HDL was not associated with P-gp expression. In addition, we did not observe any effect of P-gp expression on cellular free and esterified cholesterol content, cholesteryl ester uptake from LDL and HDL particles, or acyl-CoA:cholesterol acyltransferase activity. Therefore, we conclude that P-gp expression does not play a major role in cholesterol homeostasis in P-gp-inducible cells and that the effects of P-gp on cholesterol homeostasis previously described in drug-selected cells might result from non-P-gp pathways that were also induced by selection for drug resistance. PMID- 16215260 TI - cDNA cloning and functional expression of jerdostatin, a novel RTS-disintegrin from Trimeresurus jerdonii and a specific antagonist of the alpha1beta1 integrin. AB - Jerdostatin represents a novel RTS-containing short disintegrin cloned by reverse transcriptase-PCR from the venom gland mRNA of the Chinese Jerdons pit viper Trimeresurus jerdonii. The jerdostatins precursor cDNA contained a 333-bp open reading frame encoding a signal peptide, a pre-peptide, and a 43-amino acid disintegrin domain, whose amino acid sequence displayed 80% identity with that of the KTS-disintegrins obtustatin and viperistatin. The jerdostatin cDNA structure represents the first complete open reading frame of a short disintegrin and points to the emergence of jerdostatin from a short-coding gene. The different residues between jerdostatin and obtustatin/viperistatin are segregated within the integrin-recognition loop and the C-terminal tail. Native jerdostatin (r jerdostatin-R21) and a R21K mutant (r-jerdostatin-K21) were produced in Escherichia coli. In each case, two conformers were isolated. One-dimensional (1)H NMR showed that conformers 1 and 2 of r-jerdostatin-R21 represent, respectively, well folded and unfolded proteins. The two conformers of the wild type and the R21K mutant inhibited the adhesion of alpha(1)-K562 cells to collagen IV with IC(50) values of 180 and 703 nm, respectively. The IC(50) values of conformers 2 of r-jerdostatin-R21 and r-jerdostatin-K21 were, respectively, 5.95 and 12.5 microm. Neither r-jerdostatin-R21 nor r-jerdostatin-K21 showed inhibitory activity toward other integrins, including alpha(IIb)beta(3), alpha(v)beta(3), alpha(2)beta(1), alpha(5)beta(1), alpha(4)beta(1), alpha(6)beta(1), and alpha(9)beta(1) up to a concentration of 24 mum. Although the RTS motif appears to be more potent than KTS inhibiting the alpha(1)beta(1) integrin, r-jerdostatin-R21 is less active than the KTS-disintegrins, strongly suggesting that substitutions outside the integrin-binding motif and/or C terminal proteolytic processing are responsible for the decreased inhibitory activity. PMID- 16215262 TI - Training time and consultant practice. PMID- 16215261 TI - Functional dissection of osteoprotegerin and its interaction with receptor activator of NF-kappaB ligand. AB - The receptor activator of NF-kappaB (RANK) belongs to the neuregulin/tumor necrosis factor (TNF) receptor superfamily and is activated by RANK ligand (RANK L), a homotrimeric, TNF-like cytokine. RANK is present on the surface of osteoclast cell precursors, where its interaction with RANK-L induces their terminal differentiation into osteoclasts, thus increasing bone breakdown. The secreted, soluble receptor osteoprotegerin (OPG) interrupts this activation by binding directly to RANK-L. Therefore, osteoclast maturation (and bone homeostasis) is regulated in vivo by OPG levels of expression. We have studied the assembly state and affinity of OPG for RANK-L by sedimentation analyses and surface plasmon resonance (Biacore). Full-length, homodimeric OPG binds to RANK-L with a KD of 10 nM. OPG is also a member of the TNF receptor superfamily and contains four disulfide-rich ligand-binding domains, yet lacks a transmembrane region separating the ligand-binding region from the two death domains, as observed for other receptor family members. We showed that dimerization of OPG results from noncovalent interactions mediated by the death domains and to a lesser extent by a C-terminal heparin-binding region. In contrast, a C-terminal intermolecular disulfide bond does not contribute to the formation or stability of OPG dimers. A truncate of osteoprotegerin, containing the ligand-binding domains but lacking the dimerization domains, bound RANK-L with a KD of approximately 3 microM, indicating that monomer oligomerization for the OPG provides an increase of 3 orders of magnitude in the affinity for RANK-L. Therefore, OPG dimer formation is required for the mechanism of inhibition of the RANK-L/RANK receptor interaction. PMID- 16215263 TI - Postoperative visual loss following prone spinal surgery. PMID- 16215264 TI - Nitrous oxide anaesthesia and intraocular gases. PMID- 16215266 TI - Regional cerebral oximetry after oxygen administration. PMID- 16215267 TI - Cardiopulmonary resuscitation training: in need of some critical care? PMID- 16215265 TI - Ketamine for treatment of catheter-related bladder discomfort. PMID- 16215268 TI - Doctors' drinking and fitness to practise. PMID- 16215271 TI - Defense and resistance-inducing activities in tobacco of the sulfated beta-1,3 glucan PS3 and its synergistic activities with the unsulfated molecule. AB - Laminarin, a beta-1,3 glucan with single beta-glucose branches at position 6, was chemically sulfated to produce PS3 with a degree of sulfation of 2.4. PS3 has previously been shown to activate the salicylic acid (SA) signaling pathway in infiltrated tobacco and Arabidopsis thaliana leaf tissues. Here, we investigated whether PS3 induces systemic defense and resistance responses in tobacco. Using a radiolabeled compound, it was first demonstrated that PS3 remains strictly localized to the infiltrated tissues. PS3 is also resistant to beta-glucanase degradation. In transgenic PR1-beta-glucuronidase (GUS) tobacco plants, PS3 causes a strong increase in GUS activity in treated tissues but none in untreated leaves. PS3-infiltrated tissues challenged with tobacco mosaic virus (TMV) 8 d after elicitor application show a decrease in both the lesion number and the lesion size, whereas treatment with laminarin, the unsulfated native glucan, affected only the lesion number. PS3 does not induce systemic acquired resistance to TMV. PS3 and laminarin show synergistic effects in promoting the oxidative burst in tobacco cell suspensions and in increasing the expression of genes encoding O-methyltransferases of the phenylpropanoid pathway in tobacco plants. No synergistic effect was observed on the expression of either the SA-dependent acidic PR1 gene or the ethylene-dependent basic PR5 gene in tobacco plants. PMID- 16215269 TI - Use of laboratory markers and the audit questionnaire by primary care physicians to detect alcohol abuse by patients. AB - AIMS: To evaluate how often laboratory markers [Mean corpuscular volume (MCV), Gamma-glutamyl transferase, Aspartate aminotransferase, Alanine aminotransferase, or Carbohydrate-deficient transferrin (CDT)] and the Alcohol Use Disorders Identification Test (AUDIT) are used to detect alcohol abuse in primary health care. METHODS: Cross-sectional self-administered questionnaire survey to all 3193 primary health care physicians in Finland. Response rate was (65.7%). RESULTS: CDT was used at least occasionally by 43.4% of the physicians. Corresponding figures were 53.4% for conventional alcohol laboratory markers (MCV, transaminases) and 67.0% for AUDIT. Almost all the respondents used some laboratory marker to detect alcohol abuse. The use of brief alcohol intervention was associated with the greater likelihood that a physician uses different methods to detect alcohol abuse. The data also indicates that gender, age, and having a specialist licence influence activity in using different methods. CONCLUSIONS: Considering the ambivalences in relation to alcohol issues in health care, the use of CDT and AUDIT are reasonably frequent. This may indicate that tools to facilitate the work may also help in adapting new activities. PMID- 16215272 TI - Hahb-10, a sunflower homeobox-leucine zipper gene, is regulated by light quality and quantity, and promotes early flowering when expressed in Arabidopsis. AB - Homeodomain-leucine zipper proteins constitute a family of transcription factors found only in plants. Expression patterns of the sunflower homeobox-leucine zipper gene Hahb-10 (Helianthus annuus homeobox-10), that belongs to the HD-Zip II subfamily, were analysed. Northern blots showed that Hahb-10 is expressed primarily in mature leaves, although expression is clearly detectable in younger leaves and also in stems. Considerably higher expression levels were detected in etiolated seedlings compared with light-grown seedlings. Induction of Hahb-10 expression was observed when seedlings were subjected to treatment with gibberellins. Transgenic Arabidopsis thaliana plants that express Hahb-10 under the 35S cauliflower mosaic virus promoter show special phenotypic characteristics such as darker cotyledons and planar leaves. A reduction in the life cycle of about 25% allowing earlier seed collection was also observed, and this phenomenon is clearly related to a shortened flowering time. When the number of plants per pot increased, the difference in developmental rate between transgenic and non transformed individuals became larger. After gibberellin treatment, the relative difference in life cycle duration was considerably reduced. Several light regulated genes have been tested as possible target genes of Hahb-10. One of them, PsbS, shows a different response to illumination conditions in transgenic plants compared with the response in wild-type plants while the other genes behave similarly in both genotypes. We propose that Hahb-10 functions in a signalling cascade(s) that control(s) plant responses to light quality and quantity, and may also be involved in gibberellin transduction pathways. PMID- 16215273 TI - Proteomic analysis of lysosomal acid hydrolases secreted by osteoclasts: implications for lytic enzyme transport and bone metabolism. AB - Osteoclasts, the bone-digesting cells, are polarized cells that secrete acid hydrolases into a resorption lacuna where bone degradation takes place. The molecular mechanisms underlying this process are poorly understood. To analyze the nature of acid hydrolases secreted by osteoclasts, we used the mouse myeloid Raw 264.7 cell line that differentiates in vitro into mature osteoclasts in the presence of the receptor activator of NF-kappaB ligand. Upon differentiation, we observed a strong increase in the secretion of mannose 6-phosphate-containing acid hydrolases. A proteomic analysis of the secreted proteins captured on a mannose 6-phosphate receptor affinity column revealed 58 different proteins belonging to several families of acid hydrolases of which 16 are clearly involved in bone homeostasis. Moreover these acid hydrolases were secreted as proproteins. The expression of most of the identified acid hydrolases is unchanged during osteoclastogenesis. Thus, our data strongly support the notion that the polarized secretion of acid hydrolases by osteoclasts results from a reorganization of key steps of membrane traffic along the lysosomal pathway rather than from a fusion of lysosomes with the membrane facing the resorption lacuna. PMID- 16215274 TI - Biomarker discovery from pancreatic cancer secretome using a differential proteomic approach. AB - Quantitative proteomics can be used as a screening tool for identification of differentially expressed proteins as potential biomarkers for cancers. Candidate biomarkers from such studies can subsequently be tested using other techniques for use in early detection of cancers. Here we demonstrate the use of stable isotope labeling with amino acids in cell culture (SILAC) method to compare the secreted proteins (secretome) from pancreatic cancer-derived cells with that from non-neoplastic pancreatic ductal cells. We identified 145 differentially secreted proteins (>1.5-fold change), several of which were previously reported as either up-regulated (e.g. cathepsin D, macrophage colony stimulation factor, and fibronectin receptor) or down-regulated (e.g. profilin 1 and IGFBP-7) proteins in pancreatic cancer, confirming the validity of our approach. In addition, we identified several proteins that have not been correlated previously with pancreatic cancer including perlecan (HSPG2), CD9 antigen, fibronectin receptor (integrin beta1), and a novel cytokine designated as predicted osteoblast protein (FAM3C). The differential expression of a subset of these novel proteins was validated by Western blot analysis. In addition, overexpression of several proteins not described previously to be elevated in human pancreatic cancer (CD9, perlecan, SDF4, apoE, and fibronectin receptor) was confirmed by immunohistochemical labeling using pancreatic cancer tissue microarrays suggesting that these could be further pursued as potential biomarkers. Lastly the protein expression data from SILAC were compared with mRNA expression data obtained using gene expression microarrays for the two cell lines (Panc1 and human pancreatic duct epithelial), and a correlation coefficient (r) of 0.28 was obtained, confirming previously reported poor associations between RNA and protein expression studies. PMID- 16215275 TI - Multiple signals regulate axon regeneration through the Nogo receptor complex. AB - Several myelin-derived proteins have been identified as components of central nervous system (CNS) myelin, which prevents axonal regeneration in the adult vertebrate CNS. The discovery of the receptor for these proteins was a major step toward understanding the failure of axon regeneration. The receptor complex consists of at least three elements: the p75 receptor (p75NTR), the Nogo receptor and LINGO-1. Downstream from the receptor complex, RhoA activation has been shown to be a key element of the signaling mechanism of these proteins. Rho activation arrests axon growth, and blocking Rho activation promotes axon regeneration in vivo. Recent studies have identified conventional protein kinase C as an additional necessary component for axon growth inhibition. Possible crosstalk downstream of these signals should be explored to clarify all the inhibitory signals and may provide an efficient molecular target against injuries to the CNS. PMID- 16215276 TI - Neuronal homeostasis through translational control. AB - Translational repression is a key component of the mechanism that establishes segment polarity during early embryonic development in the fruitfly Drosophila melanogaster. Two proteins, Pumilio (Pum) and Nanos, block the translation of hunchback messenger RNA in only the posterior segments, thereby promoting an abdominal fate. More recent studies focusing on postembryonic neuronal function have shown that Pum is also integral to numerous mechanisms that allow neurons to adapt to the changing requirements placed on them in a dynamic nervous system. These mechanisms include those contributing to dendritic structure, synaptic growth, neuronal excitability, and formation of long-term memory. This article describes these new studies and highlights the role of translational repression in regulation of neuronal processes that compensate for change. PMID- 16215277 TI - Removing pathogenic memories: a neurobiology of psychotherapy. AB - Experimental research examining the neural bases of nondeclarative memory has offered intriguing insight into how functional and dysfunctional implicit learning affects the brain. Long-term modifications of synaptic transmission, in particular, are currently considered the most plausible mechanism underlying memory trace encoding and compulsions, addiction, anxiety, and phobias. Therefore, an effective psychotherapy must be directed to erase maladaptive implicit memories and aberrant synaptic plasticity. This article describes the neurobiological bases of pathogenic memory disruption to provide some insight into how psychotherapy works. At least two mechanisms of unwanted memory erasing appear to be implicated in the effects of psychotherapy: inhibition of memory consolidation/reconsolidation and extinction. Behavioral evidence demonstrated that these two ways to forget are profoundly distinct in nature, and it is increasingly clear that their cellular, synaptic, and molecular underpinnings are different. Accordingly, the blockade of consolidation/reconsolidation erases memories by reversing the plasticity associated with memory maintenance, whereas extinction is a totally new form of plasticity that, similar to the plasticity underlying the old memory, requires protein synthesis-dependent synaptic remodeling. PMID- 16215279 TI - AMPA receptor subunit GluR2 gates injurious signals in ischemic stroke. AB - Ischemic stroke, or a brain attack, is the third leading cause of death in developed countries. A critical feature of the disease is a highly selective pattern of neuronal loss; certain identifiable subsets of neurons--particularly CA1 pyramidal neurons in the hippocampus are severely damaged, whereas others remain intact. A key step in this selective neuronal injury is Ca2+/Zn2+ entry into vulnerable neurons through alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor channels, a principle subtype of glutamate receptors. AMPA receptor channels are assembled from glutamate receptor (GluR)1, 2, -3, and -4 subunits. Circumstance data have indicated that the GluR2 subunits dictate Ca2+/Zn2+ permeability of AMPA receptor channels and gate injurious Ca2+/Zn2+ signals in vulnerable neurons. Therefore, targeting to the AMPA receptor subunit GluR2 can be considered a practical strategy for stroke therapy. PMID- 16215278 TI - Interactions of interleukin-1 with neurotrophic factors in the central nervous system: beneficial or detrimental? AB - Interleukin (IL)-1 is a multifunctional cytokine that plays a key role in mediating inflammation in the brain. Many different cell types in the brain express the IL-1 receptor and respond to this cytokine by activating cell-type specific signaling pathways leading to distinct functional responses, which collectively comprise the inflammatory response in the brain. One key effect of IL-1 in the brain is the induction of trophic factor production by glial cells, which has traditionally been considered a neuroprotective response to injury or disease. However, recent studies have shown that nerve growth factor, which is regulated by IL-1, can induce neuronal survival or apoptosis via different receptors. This article examines the interaction of IL-1 with different trophic factors in the brain. PMID- 16215280 TI - bHLH genes and retinal cell fate specification. AB - The various cell types in the vertebrate retina arise from a pool of common progenitors. The way that the cell types are specified has been a long-standing issue. Decades of research have yielded a large body of information regarding the involvement of extrinsic factors, and only recently has the function of intrinsic factors begun to emerge. This article reviews recent studies addressing the role of basic helix-loop-helix (bHLH) factors in specifying retinal cell types, with an emphasis on bHLHhierarchies leading to photoreceptor production. Photoreceptor genesis appears to employ two transcriptional pathways: ngn2-->neuroD-->raxL and ath5-->neuroD-->raxL. ngn2 and ath5 function in progenitors, which can potentially develop into different cell types. neuroD represents one of the central steps in photoreceptor specification. Ath5 is also essential for ganglion cell development. It remains to be demonstrated whether a bHLH gene functions as a key player in specifying the other types of retinal cells. Genetic knockout studies have indicated intricate cross-regulation among bHLH genes. Future studies are expected to unveil the mechanism by which bHLH factors network with intrinsic factors and communicate with extrinsic factors to ensure a balanced production of the various types of retinal cells. PMID- 16215282 TI - Proceedings from the 1st Seminar on Cardiovascular Engineering: Medical Engineering and Therapy. 27-28 April 2004, Nancy, France. PMID- 16215283 TI - Regulation of cell adhesion. AB - Cell function usually requires an accurate control of attachment to and detachment from many other cells or biological surfaces. This is usually achieved by a combination of multiple cell processes the relative importance of which may be difficult to assess. The aim of this review is to discuss the role of different mechanisms used to control adhesion on the basis of selected examples and recently developed methodologies allowing quantitative study of cell adhesion. It is concluded that cells control adhesion by modifying (i) adhesion receptor expression, as a consequence of exocytosis, endocytosis, or proteolytic mechanisms, (ii) adhesion receptor intrinsic activity, through a variety of conformational changes, (iii) receptor organisation in cell membranes, as a consequence of topographical distribution and clustering, lateral mobility, and strength of anchoring to the cytoskeleton, and (iv) general processes unrelated to a specific receptors, such as glycocalyx changes or modification of cell shape or surface mechanical properties. PMID- 16215284 TI - Biomechanical aspects in tissue engineering. AB - This article provides a very brief overview of how some major mechanical properties of biological tissue and their substitutes can be described and quantified in basic form in order to understand their physiological functioning from the viewpoint of tissue regeneration, then allowing new developments in tissue engineering. After shortly reviewing the main rheological properties we have focused on the related phenomena of mass and momentum transport through tissue, considering the poroelastic characteristics of these media. Using very rough approach, it is shown how the biphasic nature of these media can influence mechanical stresses and nutriment feeding of the imbedded cells. PMID- 16215285 TI - Functional tissue engineering of the aortic heart valve. PMID- 16215286 TI - Roles of mechanical blood forces in vascular diseases. A clinical overview. AB - Under the influence of heart and elasticity of arteries, circulating blood constantly acts upon the layers of the vessels, covered by active endothelial cells. The shear stress appears like the most efficient mechanical factor developing a rubbing physical force, the laminar flow. But pulsating and centrifugal forces allow shear to be often unsteady. Mechanobiology investigates receptors and transduction across wall cells, showing that thousands of genes are activated in the endothelium leading to a lot of adaptable functions. Atherosclerosis is due to disturbances of laminar flow in specific areas where low shear allows white cells to adhere and migrate, permeability to increase. Coronaropathies, cerebral vascular accidents, aneurisms are so geometrically local diseases, of which lesions are subsequently enhanced by risk factors. Chronic venous insufficiency is also related to physical forces: hydrostatic and centrifugal pressures, and disruption of shear stress along the wall and around valvulae. Here also similar cells and biochemical phenomenons are the cause of wall remodelling and varicose. At the level of microcirculation the shear stress fall induces hypoxia, accumulation of white cells and hemorheological disorders in microvenulae. This leads to lesions of tissue, small vessel and ulcers. Numerous treatments in vascular diseases tend to restore flow and blood shearing. However a better understanding in the future should open new therapeutic fields and genetic approaches. PMID- 16215287 TI - Single stage cell seeding of small diameter prosthetic cardiovascular grafts. AB - Small-diameter prosthetic cardiovascular bypass grafts have high occlusion rates. Thrombogenicity caused by the lack of endothelial cells (ECs) on the luminal surface of the grafts is one of the main reasons for its occlusion. One strategy to improve the clinical performance of cardiovascular prosthetic grafts has been to seed its luminal surface with a monolayer of the patient's own ECs. In this strategy a "two stage" seeding procedure is utilized whereby cells obtained from a vein are amplified in cell culture, then seeded onto a fibrin-arginine-glycine aspartate (RGD) tripeptide-enriched expanded polytetrafluoroethylene (ePTFE) graft in a rotating bioreactor for one week, after which it is surgically implanted. This achieves patency rates approaching those of vein grafts. The disadvantage of two stage seeding is that it requires culture facilities, a large amount of RGD, which is expensive and is confined to elective cases because of the delay between cell cultivation, seeding, and graft implantation. A single stage seeding using freshly extracted ECs that is transplanted onto the graft at the same time frame of the bypass operation without the need for cell cultivation would be an ideal answer for the disadvantages of two stage seeding. Animal trials have been successful but human trials of single stage seeding have been disappointing. It has been hypothesized that extracted ECs are scarce, furthermore, they are washed off the graft surface once exposed to blood flow. This review examines the various techniques/technologies to improve endothelial cell extraction from various sources and retention onto the luminal surface of prosthetic cardiovascular grafts in order to develop a clinically applicable strategy for single stage seeding. PMID- 16215281 TI - Mood stabilizers target cellular plasticity and resilience cascades: implications for the development of novel therapeutics. AB - Bipolar disorder is a devastating disease with a lifetime incidence of about 1% in the general population. Suicide is the cause of death in 10 to 15% of patients and in addition to suicide, mood disorders are associated with many other harmful health effects. Mood stabilizers are medications used to treat bipolar disorder. In addition to their therapeutic effects for the treatment of acute manic episodes, mood stabilizers are useful as prophylaxis against future episodes and as adjunctive antidepressant medications. The most established and investigated mood-stabilizing drugs are lithium and valproate but other anticonvulsants (such as carbamazepine and lamotrigine) and antipsychotics are also considered as mood stabilizers. Despite the efficacy of these diverse medications, their mechanisms of action remain, to a great extent, unknown. Lithium's inhibition of some enzymes, such as inositol monophosphatase and glycogen synthase kinase-3, probably results in its mood-stabilizing effects. Valproate may share its anticonvulsant target with its mood-stabilizing target or may act through other mechanisms. It has been shown that lithium, valproate, and/or carbamazepine regulate numerous factors involved in cell survival pathways, including cyclic adenine monophospate response element-binding protein, brain-derived neurotrophic factor, bcl-2, and mitogen-activated protein kinases. These drugs have been suggested to have neurotrophic and neuroprotective properties that ameliorate impairments of cellular plasticity and resilience underlying the pathophysiology of mood disorders. This article also discusses approaches to develop novel treatments specifically for bipolar disorder. PMID- 16215288 TI - In vitro endothelialized ePTFE prostheses: clinical update 20 years after the first realization. AB - The replacement of arteries with synthetic vascular prostheses often leads to failure when small-diameter or low-flow locations are concerned, due in part to the thrombogenicity of the graft surface. In order to improve long-term patency of these grafts, the concept of endothelial cell seeding has been suggested, the composite structure resulting from the combination of biologically active cells to prosthetic materials thus creating more biocompatible vascular substitutes. To achieve endothelialization of synthetic grafts, previous efforts aimed at "one stage" procedure in the 1980's seemed clinically feasible but results of reported clinical trials were controversial and mostly disappointing. An alternative method is an in vitro complete and preformed endothelial lining at the time of implantation: the "two-stage" procedure which implies harvest and culture of autologous endothelial cells. Up to date, the latter approach demonstrated its superiority in terms of significantly increased patency of the grafts that underwent endothelialization several years earlier. PMID- 16215289 TI - Experimental comparison between autofluorescence spectra of constrained fresh and cryopreserved arteries. AB - The study of mechanical properties of the arterial wall is an important step in the comprehension of the vascular physiopathological functioning. However, cryopreserving biological tissues using very low temperatures can induce biological and structural modifications which may involve complications (dilatation, bursting, stenosis) after reimplantation. Many procedures of mechanical tests (traction, dilatation) developed in research allow us to comprehend and analyse rheological behaviour of the arterial wall. The study presented in this article offers a new perspective to detect changes of mechanical properties of cryopreserved arterial samples. In fact, the original idea is to couple a mechanical test bed (uniaxial traction of arterial rings) with spectroscopic measurements (autofluorescence) for the purpose of correlating mechanical modifications and spectral variations. Ultimately, this new approach could lead to develop a device allowing atraumatic and contactless optical examinations of arterial graft to determine its mechanical state before reimplantation. PMID- 16215290 TI - Kinetics of the endocytotic pathway of Low Density Lipoprotein (LDL) in human endothelial cells line under shear stress: an in vitro confocal microscopy study. AB - We studied the effect of mechanical forces (shear stress) on the kinetics of internalization of native LDL and ox-LDL in endothelial cell line ECV304. This study was performed by using Confocal microscopy and FRET with two carbocyanine dyes, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiO) as the donor and 3,3'-dioctadecyloxacarbocyanine perchlorate (DiI) as the receptor. The cells were incubated with a culture medium containing either 10 microg/ml DiI LDL or DiO-LDL in static conditions or subjected to a laminar flow under a Confocal Laser Scanning Microscope (SP2 Leica, Germany). The results showed: (1) the possibility to evaluate the kinetics of LDL endocytosis in living cells, (2) shear stress in comparison with control group more effectively enhanced LDL uptake, (3) ox-LDL (>50 microg/ml) >4 hours incubation was found to affect the cells as reflected by their detachment at low shear stress. PMID- 16215291 TI - Role of Ca2+ in the effects of shear stress and TNF-alpha on caveolin-1 expression. AB - The aim of this work was to study the influences of Ca2+ (medium free calcium, with BAPTA, with 100 mM Ca2+, 100 mM Ca2++10 microM ionomycin) on the expression of caveolin-1 (structural protein of caveolae) of endothelial cells (ECs) submitted to mechanical forces (shear stress) or biochemical stimulation (TNF alpha). We found that shear stress enhanced the caveolin-1 expression. Simultaneously, the caveolin-1 expression is dependant on [Ca2+]i: [Ca2+]free medium+100 microM BAPTA<[Ca2+]free medium<[Ca2+]100 mM<[Ca2+]100 mM+10 microM ionomycin. In contrast, TNF-alpha induced a decrease of caveolin-1 in the cells, whatever the [Ca2+]i. These results suggest that there could be a synergistic effect between shear stress and Ca2+ on caveolin-1 expression but an inflammatory stimulation (TNF-alpha) induces a down regulation of caveolin-1 expression. PMID- 16215292 TI - Autologous cell based therapy for treating chronic infarct myocardium. AB - Recent experimental and clinical studies have shown that autologous cell based therapy using skeletal myoblasts or bone marrow-derived stem cells might have beneficial effects in chronic ischemic heart disease. The underlying concept is based on the repopulation of necrotic tissue by either readily contractile myoblasts or by bone marrow-derived stem cells. However, there is a need to resolve a number of issues for determining the better way to perform these treatments and, moreover, for assessing the real beneficial functional effect of each of these cell therapies. In this mini-review, we will discuss (i) the issues of the selection of chronic infarct animal to truly determine the impact of cell therapy on cardiac function recovery, and (ii) the evaluation of the bio availability and the bio-distribution of transplanted cells. Some new investigational methodologies based on clinical end-points are also proposed. PMID- 16215293 TI - Behaviour of endothelial cells seeded on thin polyelectrolyte multilayered films: a new biological scaffold. AB - The surface modification using thin polyelectrolyte multilayered films was proposed as a new scaffold material for different cell lines. In this study, we evaluated the possible use of polyelectrolyte multilayers as surface modification for the development of endothelial cells. In order to control the behaviour of endothelial cells, cell viability by MTT assay was studied. Moreover, the endothelial cell phenotype was checked and the expression of a leukocyte adhesion molecule (ICAM-1) was quantified. The behaviour of the cells on two polyelectrolyte multilayers was compared to cells on polystyrene, and two polyelectrolyte monolayers (terminating the multilayer architectures). The results have shown a better cell viability on the polyelectrolyte multilayers, inducing a higher cell number compared to polyelectrolyte monolayers after 1 and 3 days of culture. Moreover, the cells showed a normal morphology of cytoskeleton. The phenotype of the endothelial cells was kept and a low level of leukocyte adhesion molecules was observed. In conclusion, the polyelectrolyte multilayers can be considered as a potential surface modification procedure to enhance the development of endothelial cells on hydrophobic substrate and which can be applied to vascular tissue engineering. PMID- 16215294 TI - Apical rigidity of an epithelial cell monolayer evaluated by magnetic twisting cytometry: ICAM-1 versus integrin linkages to F-actin structure. AB - Using Magnetic Twisting Cytometry (MTC) technique, we attempted to characterize in vitro the rigidity of the lining tissue covering the lung alveolar wall from its apical face. We purposely used a cellular model constituted by a monolayer of human alveolar epithelial cell (A549) over which microbeads, fixed to InterCellular Adhesion Molecule (ICAM-1), exert a controlled mechanical stress. ICAM-1 expression was induced by Tumor Necrosis Factor-alpha (TNF-alpha). Rigidity measurements, performed in the course of cytochalasin D depolymerization, reveal the force transmitter role of the transmembrane receptor ICAM-1 and demonstrate that ICAM-1 and F-actin linkages confers mechanical rigidity to the apical face of the epithelial cell monolayer resembling that provided by integrins. These results confirm the ability of MTC in identifying transmembrane mechanoreceptors in relation with F-actin. Molecular linkages between ICAM-1 and F-actin were observed by spatial visualisations of the structure after double staining of F-actin and anti ICAM-1 antibody through confocal microscopy. PMID- 16215295 TI - New tool to monitor membrane potential by FRET Voltage Sensitive Dye (FRET-VSD) using Spectral and Fluorescence Lifetime Imaging Microscopy (FLIM). Interest in cell engineering. AB - In this work, we investigated a voltage-sensitive fluorescent system to monitor membrane potential by spectral and lifetime fluorescence microscopy. A two component FRET sensor has been designed that utilizes fluorescent phospholipids acceptor (DHPE-TRITC) bound on one side of the membrane and donor molecules (oxonol) which are sensitive to membrane potential. We used multiphoton excitation and FLIM to deliver contrast lifetimes of different line cancerous cells. These results provide new information concerning the differential response to depolarized cancerous cells from resting cells when compared to fibroblast normal cells. Given the sensitivity and the fast time response, this FRET system may be particularly useful for applications involving compression of tissues by mechanical forces. PMID- 16215296 TI - Old prospects in cardiovascular engineering. AB - For more than thirty years the thrombogenicity of blood-contacting materials has been deplored. This impotency, combined with inappropriate mechanical properties of these materials, explains the poor patency of medium-sized arterial prostheses and the lack of substitutes for small diameter arteries (psi50% response respectively (p=0.34, Fisher's exact test). Mean PSA nadir in those who responded were 20.6 ng/ml (range 1.6-59.4) and 7.41 ng/ml (range 0.1-42.6) in groups A and B respectively. The median duration of response was 9 months (3-18 months) for group A and 12 months (3-18 months) for group B. Seven patients in group A and 6 in group B experienced adverse events. Three of the 7 in the group A experienced cardiovascular related adverse effects (1 congestive cardiac failure, 1 pulmonary embolism and 1 stroke). At the end of the study period, 14 (54%) of group A patients were alive and 12 (46%) were dead. In group B, 15 (47%) were alive, 16 (50%) were dead and 1 (3%) lost to follow-up. At the completion of the study, 3 patients in each group were still on treatment. CONCLUSION: Low-dose DES and 50 mg of bicalutamide per day are equally effective in hormone refractory prostate carcinoma with respect to biochemical response, although DES has more severe adverse effects. This is a small sample and larger multicentre trials are needed to give us a definite conclusion. PMID- 16215309 TI - Predictive value of the international prostate symptom score for positive prostate needle biopsy in the low-intermediate prostate-specific antigen range. AB - PURPOSE: Serum prostate-specific antigen (PSA) has a restricted predictive value for prostate cancer in the low-intermediate PSA range (2.5-10 ng/ml). Our aim was to determine the predictive value of the International Prostate Symptom Score (IPSS) for positive prostate needle biopsy (PNB) in patients who underwent transrectal ultrasound (TRUS)-guided prostate biopsy with a low-intermediate PSA level. PATIENTS AND METHODS: Between 2001 and 2004, the data of 389 consecutive patients applying for any urologic complaint to our department and who underwent TRUS-guided prostate biopsy due to an elevated serum PSA and/or abnormal digital rectal examination (DRE) were retrospectively analyzed. A total of 158 eligible patients with a low-intermediate PSA level were included in the study. The patient's age, PSA, free PSA, free/total PSA, prostate volume, PSA density (PSAD), pre-biopsy IPSS were compared in the positive and negative biopsy groups. RESULTS: Fifty-eight of 158 patients (37%) who underwent TRUS-guided prostate biopsy had positive PNBs. Forty-eight patients (30%) had abnormal DREs. In the positive PNB group, the mean age was older and PSAD was higher, but the means of the prostate volumes and total IPSS were lower (p<0.05). Multivariate analysis demonstrated that age and IPSS were independent predictors of a positive PNB (p<0.05). The odds ratio of mild IPSS for positive PNB controlled for age was 3.0 (95% CI 1.5-6.7). Receiver-operating characteristics analysis revealed a mild IPSS (AUC=0.640) and was a considerable predictor for positive PNB as well as PSAD (AUC=0.648). The sensitivity and specificity of IPSS with a cutoff value of 7.5 points were 31 and 87% for prediction of prostate cancer detection. CONCLUSION: In the low-intermediate PSA range, mild IPSS may be a predictive factor for positive PNB with a similar specificity of PSAD. PMID- 16215310 TI - Testicular biopsy during orchidopexy procedure: does it have an adverse effect on fertility? AB - INTRODUCTION: The aim of this study is to evaluate the effect of the gonadal biopsy procedure on fertility and histology of the experimentally created undescended testis. MATERIALS AND METHODS: Thirty Wistar-Albino rat litters were divided into three groups. In the first two groups, undescended testes were obtained by dividing the right gubernaculum in the first 48 h and the third group constituted the sham group. When the rats were 30 days old, orchidopexy was performed in the first group (O), whereas orchidopexy and testicular biopsy were performed in the second group (OB). After the fertility study all rats were sacrificed at week 11 and testicular weights, mean seminiferous tubular diameters (MSTD) and mean testicular biopsy scores (MTBS) were measured. RESULTS: One testis in the O group and three testes in the OB group were atrophied and these rats were excluded from the study. Testicular weights, MSTD and MTBS values of the operated side in the O and OB groups were lower than those in the sham group. There was a statistically significant difference between the MSTD values of the ipsilateral testes in the OB group and sham group (p<0.05). MTBS values of the ipsilateral testes in the O and OB groups were <8. There was no difference in all parameters in the contralateral testes between all groups (p>0.05). All the evaluated rats were fertile. While there was no difference in number of fetuses and fertility scores between the O and OB group, however, the differences were significant between each of these groups and the sham group (p<0.05). CONCLUSION: Testicular biopsy in the unilateral undescended testis model results in limited histological changes without affecting the fertility. PMID- 16215311 TI - Does lidocaine ointment addition increase fluoxetine efficacy in the same group of patients with premature ejaculation? AB - PURPOSE: To evaluate the efficacy of fluoxetine alone and fluoxetine+lidocaine ointment in the same patient group with premature ejaculation (PE). MATERIAL AND METHODS: 78 patients with PE were given 20 mg fluoxetine by an 'as-needed treatment' 4 h before planned sexual activity for a period of 3 months. They were then told to add local lidocaine ointment to fluoxetine 30 min before sexual activity for an additional 3 months for most of their sexual attempts. They were asked to note their PE grades and intravaginal ejaculatory latency time (IELT) scores by stopwatch technique before and after each treatment modality; the results were compared statistically afterwards. RESULTS: Of 46 patients who completed the study, the mean pretreatment, fluoxetine alone and fluoxetine+lidocaine ointment treatment PE grades and IELT scores were found to be 6.52+/-1.42 and 2.58+/-0.49, 3.21+/-1.86 and 1.28+/-0.71, 2.17+/-1.56 and 1.04+/-0.72, respectively, showing a decrease in PE grades and IELT scores in combined therapy. On an individual patient basis, the total significant and moderate improvement rate of combined therapy was found to be 86.9%. Failure was observed in 6 (13.1%) patients. CONCLUSION: The effective treatment with fluoxetine+lidocaine ointment offers the advantage of an 'as-needed treatment' in PE with minimal side effects and can be used as one of the first-line alternatives in the treatment of PE. PMID- 16215312 TI - Epididymoorchitis due to brucellosis in central Anatolia, Turkey. AB - Brucellosis may involve many organs and tissues. Epididymoorchitis is a focal genitourinary complication of human brucellosis. In this study, we describe our experience with the diagnosis, treatment, and final outcomes of 17 patients with epididymoorchitis out of 186 male patients with brucellosis between March 1999 and December 2003. The rate of epididymoorchitisdue to brucellosiswas 9.1%. All subjects complained about swollen, painful testicles. The duration of their complaint varied between 1 week and 2 months. Both testis and epididymis were involved in 15 patients and 2 had bilateral involvement. The patients were treated with medical treatment and a complete resolution was achieved in all of them. Patients with Brucella infection occasionally manifest genitourinary complications. Clinicians, especially those serving in endemic areas or serving patients coming from endemic areas, should consider the likelihood of brucellosis as a cause of epididymoorchitis. PMID- 16215313 TI - Assessment of obstruction in adult ureterocele by means of color Doppler duplex sonography. AB - AIM: The purpose of this study is to present a method for identifying a ureteral obstruction in unilateral orthotopic ureterocele by means of conventional sonography and color Doppler duplex sonography. We focus on the measurement of the ureterocele dimensions, the degree of dilation it causes to the ipsilateral upper urinary tract, the registration of urine out-flow from the ureteral orifice into the bladder and its spectral analysis. MATERIAL AND METHOD: Over 2 years at our institutions, 8 adult patients (7 women, 1 man) were diagnosed as having a single system orthotopic ureterocele. Four of them presented with lumbar pain, dysuria and recurrent urinary tract infections, while the remainder were asymptomatic and diagnosed accidentally. The diagnosis was based on serial sonography of the upper and lower urinary tract confirmed by intravenous pyelography and cystoscopy. We also performed color Doppler duplex sonographic evaluation of the urine jets ejected from both ureteral orifices into the bladder. Using the flow spectral study we analyzed the waveforms and measured their duration and flow rate. The study was completed with a comparative analysis of the data obtained from both ureteral orifices. RESULTS: Cystic dilation of the lower ureteric extremity into the bladder was presented in all cases. Upper urinary tract dilation, of various grades, was present in 4 of 8 patients. Differences in urine jets between those derived from the ureterocele and those from the healthy contralateral ureteral orifice were significant in those patients with dilation of the upper urinary tract. The differences concerned mainly the frequency and symmetry of the jets as well as the pattern, duration and velocity of their waves. The 4 above-mentioned patients, with dilated upper urinary tracts and waveforms differentiated from the contralateral ones, were characterized as obstructive. On the other hand, the remaining 4 patients with subclinical ureterocele showed insignificant differences in urine jets and waveforms, and were found to be non-obstructive. CONCLUSION: Conventional sonography of the urinary tract in combination with color Doppler duplex sonography of the ureteral jets can be used in an attempt to diagnose and evaluate a unilateral orthotopic (single system) ureterocele and assess the necessity of intervention to identify the obstruction. PMID- 16215314 TI - Effectiveness of urethral injection of botulinum A toxin in the treatment of voiding dysfunction after radical hysterectomy. AB - OBJECTIVE: After radical hysterectomy for cervical cancer patients may have difficulty in urination due to detrusor underactivity and a non-relaxing urethral sphincter. This study evaluated the effectiveness of urethral injection of botulinum A toxin in treating voiding dysfunction in these patients. METHODS: Thirty patients with difficult urination after radical hysterectomy due to cervical cancer were enrolled to receive urethral injection of 100 units of botulinum A toxin (n=20) or medical treatment as controls (n=10). The clinical results and urodynamic parameters at baseline and after treatment were compared in the study group, and the quality of life (QOL) index was compared between the study and control groups. RESULTS: After urethral injection of botulinum A toxin, 8 patients had excellent results (40%) and 8 had improved results (40%) in the study group. Both voiding pressure (115.2+/-63.7 vs. 90.2+/-49.5 ml, p=0.025) and post-void residual volume (330.9+/-124.9 vs. 183.9+/-183.4 ml, p=0.011) improved significantly after treatment. The obstructive symptom score was significantly reduced (17.5+/-4.7 vs. 5.7+/-2.3 points, p=0.000) and the QOL index also improved (4.5+/-2.7 vs. 2.3+/-2.3 points, p=0.000) after treatment. The success rate was 80% in the study group. There were no significant changes in obstructive symptom scores or the QOL index in the control group. The maximal effect appeared about 1 week after treatment. The duration of the therapeutic effect ranged from 3 to 9 months. Mild stress urinary incontinence and nocturnal enuresis were noted in 7 patients (35%). CONCLUSION: Urethral injection of botulinum A toxin can be effectively used to treat patients with detrusor underactivity and non-relaxing urethral sphincter after radical hysterectomy with few adverse effects. PMID- 16215315 TI - Increased expression of EZH2, a polycomb group protein, in bladder carcinoma. AB - INTRODUCTION: Recent experiments have demonstrated that polycomb group gene enhancer zeste homolog 2 (EZH2) is highly expressed in many cancer types. Therefore, we aim to demonstrate EZH2 gene expression in transitional cell bladder cancer. PATIENTS AND METHODS: The reverse transcriptase-polymerase chain reaction (RT-PCR) was used for detection of EZH2 mRNA levels in healthy and cancerous human bladder specimens. Also, expression of the particular protein was determined by Western blotting and immunohistochemistry to confirm RT-PCR results. RESULTS: Gradually increased expression of EZH2 was detected by mRNA and protein levels in highly advanced bladder cancer specimens. In contrast, 100% of control subjects were negative for EZH2 expression. The expression of EZH2 was more frequent in G3 (92%) than G1-G2 (62-63%) and more frequent in T1-2 (72-85%) than Ta (56%). Western blot analysis results confirm the RT-PCR results. CONCLUSIONS: EZH2 overexpression precedes high frequencies of proliferation and the gradual advance of bladder cancer. These observations suggest that deregulated expression of EZH2 is associated with bladder carcinoma. PMID- 16215316 TI - Morphological responses of the rabbit testis to ischemic/reperfusion injury due to torsion. AB - OBJECTIVE: To investigate the morphological effects of free radical injury on spermatogenic cells in both testes of the rabbit experimental model of testicular torsion. MATERIALS AND METHODS: The left testes of 8 peripubertal NZW rabbits (3 6 months) were subjected to 0, 15, 30, 60, 90, 120, and 180 min of ischemia by applying a clamp to the spermatic cord, followed by reperfusion. Another set of 8 rabbits was subjected to 60 min of ischemia and administered antioxidants (acetylsalicylic acid, ascorbic acid, allopurinol, quercetin, superoxide dismutase) before reperfusion. Both testes of 4 animals per group were harvested at 24 h and the remaining 4 at 3 months. Johnsen scores for spermatogenic activity and other changes were assessed histologically and these were compared with testicular malondialdehyde (MDA), a measure of free radical damage, assayed on testicular homogenates using the thiobarbiturate method. RESULTS: In the 24 hour reperfusion group, apoptotic bodies and giant cells were more prominent in the seminiferous tubules of the left testes compared to the right, and were maximal after 90 min. In the 3-month reperfusion group, giant cells were absent, and apoptotic bodies were reduced in both testes. Testicular MDA showed an increase only in the left testes in the 24-hour reperfusion group, while the 3 month group showed increased MDA levels in both testes, but more on the left. The Johnsen score fell only to 8.0 in the left testes in the 24-hour reperfusion group, but dropped to 2.3 in the 3-month reperfusion group. Only in the 3-month reperfusion group, did antioxidant-treated animals show a fall in Johnsen scores in the left testes, regardless of the type of antioxidant. CONCLUSION: These findings confirm a role for reactive oxygen species (ROS) in damage to spermatogenic cells in both the ipsilateral and contralateral testes following torsion, with longer term effects in the torted testis. Currently available antioxidants do not provide any significant long-term protection against morphological damage to the testis by ROS generated in testicular torsion. PMID- 16215317 TI - TAP 2 gene Msp-I polymorphism might be associated with calcium oxalate stone disease. AB - INTRODUCTION: Inflammation might be one of the causes of stone disease. The function of the transporter associated with antigen-processing protein (TAP) is related to immune response and inflammation. Our aim was to investigate the relationship between stone disease and 5 polymorphic sites of the TAP gene (TAP1 1, 1-2, 2-1, 2-2, 2-3). MATERIALS AND METHODS: We compared the frequencies of 5 polymorphisms in the TAP gene between 208 patients with recurrent calcium oxalate stone and 210 healthy controls. The polymorphism was detected by polymerase chain reaction-based restriction analysis. RESULTS: Significant differences in the frequency of the polymorphism at the TAP2-2 site were detected between normal individuals and calcium stone disease patients (p<0.0001). The distribution of the genotype AA homozygote was higher in stone patients (33.3%) than in the control group (16.3%). The odds ratio for the A allele compared with the G allele was 2.097 (95% CI 1.571-2.802). CONCLUSIONS: We conclude that the TAP2-2 MspI polymorphism might be associated with calcium stone disease. PMID- 16215318 TI - Expression and functional influence of cellular retinoic acid-binding protein II in renal cell carcinoma. AB - INTRODUCTION: Retinoic acid (RA) and its derivates possess antiproliferative and tumor-suppressive abilities and are successfully used in the treatment of various malignancies. However, in metastatic renal cell carcinoma (RCC), its application did not meet first expectations. As the exact mechanisms of RA action and especially the role of the cellular retinoic acid-binding proteins (CRABP) still remain unclear, we studied the expression of CRABP-II and its potential influence on RA response in RCC. MATERIALS AND METHODS: We used the real-time RT-PCR methodology to investigate CRABP-II expression in 12 RCC samples and corresponding normal kidney tissue. Moreover, CRABP-II was cloned and overexpressed in CAKI-2 RCC cells. CRABP-II (un)transfected CAKI-2 cells were stimulated with all-trans RA (ATRA) and 9-cis RA, and their antiproliferative effects were evaluated using 3H-thymidine-proliferation assays. RESULTS: Using RPS9 and RPLP0 to normalize its expression, the median tumor/kidney ratio for CRABP-II expression was 0.16 and 0.12, respectively. Using proliferation assays, CRABP-II overexpressing CAKI-2 cells did not exhibit a significant change in RA sensitivity, but appeared to be less sensitive toward RA-stimulation compared to CAKI-2 cells expressing naturally low levels of CRABP-II (maximum difference, 59% at 3 microM ATRA). CONCLUSIONS: We were able to demonstrate a downregulation of CRABP-II expression in primary RCC tumor samples compared to the corresponding normal kidney tissue. However, CRABP-II overexpression in CAKI-2 RCC cells did not significantly influence RA associated antiproliferative actions. Further experiments are necessary to define the exact role of CRABP-II and its downregulation in RCC including its influence and dependence on other molecules involved in RA signalling and metabolism. PMID- 16215319 TI - Free insulin-like growth factor-1 levels in bladder growth following spinal cord injury experimentally. AB - INTRODUCTION: To determine the levels of spinal cord injury (SCI) on free insulin like growth factor-1 (IGF-1) content in the bladder and the possible role of free IGF-1 in growth in hypertrophic bladders in an experimental model. MATERIALS AND METHODS: The contents of free IGF-1 and protein in paraplegic and non-paraplegic rats which was induced experimentally by SCI were measured. The wet weights of the bladders were also determined. The results gained were compared. RESULTS: The mean bladder free IGF-1 level in paraplegic rats was significantly higher than the corresponding bladder free IGF-1 level in sham-operated bladder (p<0.05). Mean wet weight bladder in paraplegic rats was significantly higher than the sham operated rats (p<0.05). Paraplegic bladder had a significantly higher mean protein content than the sham-operated bladder (p<0.05). Serum free IGF-1 levels in the two groups were not different. CONCLUSION: Our pilot study reveals that free IGF-1 may be effective in SCI. PMID- 16215320 TI - Extramedullary hematopoiesis in chronic myelofibrosis encasing the pelvicaliceal system and perirenal spaces: CT findings. AB - We examined a 46-year-old male patient with idiopathic myelofibrosis diagnosed 28 months earlier. After chemotherapy and irradiation, CT showed nonnodular, soft tissue masses in the renal hilum, pelvicaliceal system, perirenal and periureteral spaces. Ultrasound-guided biopsy showed extramedullary hematopoiesis (EMH). The mass, which caused delayed nephrographic enhancement, without hydroureteronephrosis, was the only site of EMH. We report the CT findings of this uncommon EMH localization. PMID- 16215321 TI - Giant cystine stone in an infant bladder with no evidence of cystinuria--valence of possible pathomechanisms. AB - Bladder calculi account for approximately 5% of all urinary calculi. Children are at high risk in endemic areas whereas otherwise the entity is rare. Diet, voiding dysfunction and uncorrected anatomical abnormalities predispose to bladder stone formation. We report the case of a 2-year-old girl with a giant cystine bladder stone without relevant cystinuria and a small left kidney. Suprapubic cystolithotomy was performed for stone removal. We discuss possible associations of cystine calculi with unilateral genitourinary pathologies in the absence of persistent excess cystinuria. PMID- 16215322 TI - Does the prostate really need androgens to grow? Transurethral resection of the prostate in a male-to-female transsexual 25 years after sex-changing operation. AB - We present the case of a male-to-female transsexual presenting with obstructive voiding symptoms due to benign prostatic hyperplasia 25 years after sex-changing operation and under continuous estrogen therapy. PMID- 16215323 TI - Clinical and subclinical ACTH-independent macronodular adrenal hyperplasia and aberrant hormone receptors. AB - ACTH-independent macronodular adrenal hyperplasia (AIMAH) is a very rare cause of endogenous Cushing's syndrome (CS). In this review, the clinical characteristics, the pathophysiology, and the management of AIMAH are described. AIMAH typically presents with overt CS, but subclinical oversecretion of cortisol has been increasingly described. The diagnosis is suspected by adrenal nodular enlargement on conventional imaging following the demonstration of ACTH-independent hypercortisolism. Final diagnosis is established by histological examination of the adrenal tissue. Bilateral adrenalectomy is the treatment of choice but unilateral adrenalectomy has been proposed in selected cases. In patients with subclinical CS, the decision to treat should be individualized. The pathophysiology of this condition has begun to be elucidated in recent years. Diverse aberrant membrane-bound receptors expressed in a non-mutated form in the adrenal gland have been found to be implicated in the regulation of steroidogenesis in AIMAH. When systematically screened, most patients with AIMAH and CS or subclinical CS exhibit an in vivo aberrant cortisol response to one or various ligands suggesting the presence of aberrant adrenal receptors. A protocol designed to screen patients for the presence of these aberrant receptors should be undertaken in all patients with AIMAH. The identification of these receptors provides the potential for novel pharmacological therapies by suppressing the endogenous ligands or blocking the receptor with specific antagonists. PMID- 16215324 TI - Food-additive-induced urticaria: a survey of 838 patients with recurrent chronic idiopathic urticaria. AB - BACKGROUND: Recurrent chronic idiopathic urticaria (RCIU) is a common skin condition that affects 0.1-3% of the population in the USA and Europe and accounts for nearly 75% of all 'ordinary' chronic urticaria (CU) cases. METHODS: We studied 838 consecutive patients with RCIU referred to hospital between 1998 and 2003. Patients with known causes of CU were excluded. Clinical history, physical examination, and symptom diaries were evaluated during two periods, a diet-free period (1 week) and a food-additive-free diet (FAFD) period (4 weeks), respectively, and two double-blind placebo-controlled (DBPC) challenges of six food additives were administered. The first DBPC challenge included a mixture of the six food additives (DBPCmixed) given to all patients. The second DBPC challenge comprised the single food additives, administered at increasing doses (DBPCsingle) to patients with a positive DBPCmixed test and 105 patients with a negative DBPCmixed test, as a control. RESULTS: The DBPCmixed challenge was positive in 116 patients. None of the 105 control patients had a positive DBPCsingle test. Only 31 DBPCsingle tests were positive in patients with positive DBPCmixed challenge. Twenty-four of the 116 patients showing a positive DBPCmixed challenge also had a positive DBPCsingle result. CONCLUSIONS: Our results confirmed that food additive hypersensitivity reactions occurred in few RCIU patients using DBPCsingle challenge. The combination of the results of FAFD and DBPCmixed challenge seems to be of considerable practical interest for allergists, internists and dermatologists, rather than the data of clinical history and the results of DBPCsingle challenge, in patients with RCIU. PMID- 16215325 TI - Occupational asthma to carmine in a butcher. AB - Hypersensitivity to carmine (E120) has been identified as a cause of food intolerance and occupational asthma. We present a case of occupational asthma following exposure to carmine in a manufacturer of sausages and review the literature. CASE REPORT: A 42-year-old non-atopic male presented with a 5-year history of rhinoconjunctivitis and asthma on occupational exposure to food additive dusts. Symptoms increased after work. The patient had been exposed for more than 20 years. METHODS: Skin prick tests were performed with a battery of common inhalant allergens and spices. Cochineal, carmine lake and additive mixes used by the patient were extracted and subsequently used for skin prick test, bronchial provocation and in vitro measurements (specific IgE, Western blot and chromatographic fractionation). RESULTS: Prick tests were positive to carmine and carmine-containing additives; carmine-specific IgE and bronchial challenge tests were also positive (PC20 = 0.0004 mg/ml and 1.6 kU/l). Western blot showed IgE binding to bands of about 30 kDa on cochineal extract and a diffuse pattern at 40 97 kDa on carmine. This result was confirmed by gel filtration chromatography and dot blot. Carmine completely inhibited IgE binding to cochineal extract. DISCUSSION: Carmine is a potential sensitizer in an occupational setting: 18 cases of occupational asthma have been described to date. Carmine allergens are poorly defined; in general, proteins from cochineal not removed by the extraction process are considered as the main allergens in carmine. Our results are consistent with this, but show that these proteins may be subject to chemical modification. PMID- 16215326 TI - Epistatic effect of TLR4 and IL4 genes on the risk of asthma in females. AB - BACKGROUND: Many studies have demonstrated a connection between asthma and T-cell cytokine genes, such as genes coding for interleukin-4 (IL4) and IL-13, which are involved in the regulation of the TH1/TH2 balance. The toll-like receptor 4 (TLR4), the principal receptor for bacterial endotoxin, has attracted attention as a potential risk factor for asthma. We examined whether the polymorphisms of the TLR4 (A/G at +896) and IL4 (C/T at -590) showed an epistatic effect on the risk of asthma or atopy. METHODS: Gene polymorphism analyses and skin prick tests were performed on asthmatic and nonasthmatic adult subjects of a Finnish population-based case-control study. The phenotype studied was persistent asthma. RESULTS: The results showed that genotypes of neither the TLR4 SNP at +896 nor IL4 SNP at -590 were separately found to be associated with asthma. However, the female carriers of allele G (i.e. genotype AG or GG) of TLR4 and allele T (genotype CT or TT) of IL4 had a significantly increased risk for asthma. No association of these genes and atopy was found. CONCLUSIONS: Our results indicate that in females the TLR4 and IL4 genes show an epistatic effect on the risk of asthma. The low LPS-responsive allele G of TLR4 and high IgE production allele T of IL4 were found to be the predisposing combination. However, there was no epistatic effect on the risk of atopy. PMID- 16215327 TI - Different profiles of wheat antigens are recognised by patients suffering from coeliac disease and IgE-mediated food allergy. AB - BACKGROUND: Dietary intake of wheat can cause two distinct immunologically mediated diseases with severe gastrointestinal manifestations, coeliac disease (CD) and IgE-mediated food allergy. The pathomechanisms underlying these diseases are different, but the profile of the target antigens in wheat has not been compared for the two diseases. METHODS: We compared IgA- and IgE-reactive antigens in wheat using sera from patients with coeliac disease (n = 35) and food allergy to wheat (n = 16) by one- and two-dimensional immunoblotting. Furthermore, the IgG subclass (IgG1-IgG4) reactivity to wheat antigens was studied by enzyme-linked immunosorbent assay. RESULTS: IgA antibodies from CD patients and IgE antibodies from allergic patients recognised distinct profiles of wheat antigens. Furthermore, the IgG subclass responses to wheat antigens were different in CD and wheat-allergic patients. CONCLUSION: This study thus demonstrates that wheat contains antigens/epitopes which are preferentially recognised by CD patients, whereas others elicit IgE-mediated food allergy. This finding suggests that the nature of a food antigen may influence the quality of the pathological immune response in the gut and has implications for the diagnosis and therapy of hypersensitivity to wheat. PMID- 16215328 TI - Naturally occurring parainfluenza virus 3 infection in adults induces mild exacerbation of asthma associated with increased sputum concentrations of cysteinyl leukotrienes. AB - BACKGROUND: Viral respiratory tract infections represent the most frequent cause of asthma exacerbation in both children and adults, but the precise mechanism of such exacerbation remains unknown. OBJECTIVES: To determine the critical mediator of naturally occurring parainfluenza virus (PIV) 3-induced mild asthma exacerbations in adults. METHODS: The study subjects were 19 adult asthmatics with mild asthma exacerbation (peak expiratory flow = 60-80% of predicted before bronchodilator use and >80% of predicted after initial bronchodilator treatment). Differential cell counts and concentrations of inflammatory markers including eosinophil cationic protein (ECP), cysteinyl leukotrienes (cysLTs), interleukin (IL)-5, IL-10 and IL-12 were measured in the induced sputum obtained from adults with PIV3- (n = 9) and non-cold-induced (n = 10) exacerbation of asthma during both acute and convalescent phases. RESULTS: PIV3 infection was confirmed by the presence of viral RNA in nasopharyngeal aspirates. Mild exacerbation of asthma was not associated with significant changes in sputum differential cell counts. Concentrations of sputum ECP and cytokines were comparable between PIV3 and non cold-induced patients. In contrast, PIV3 infection was associated with a significant increase in sputum cysLTs during the acute phase of mild asthma exacerbation. CONCLUSIONS: Our results identified cysLTs as a critical mediator of PIV3-induced acute asthma exacerbation. PMID- 16215329 TI - Prenatal 2-dimensional and 3-dimensional ultrasonography diagnosis and autoptic findings of isolated ectopia cordis. AB - Ectopia cordis is a very rare congenital malformation, commonly associated with intracardiac anomalies. It is due to a defect in fusion of the anterior chest wall resulting in an extrathoracic location of the heart. We report prenatal 2 dimensional (2D) and 3D ultrasonography diagnosis and postnatal autoptic findings of an isolated ectopia cordis with tricuspid atresia. Ectopia cordis prenatal diagnosis is easily made with ultrasound by visualizing the heart outside the thoracic cavity. 3D ultrasonography may add more detailed visualization of the heart anomaly even if the 2D ultrasonography alone permits the prenatal diagnosis. Obstetrical management should include a careful search for associated anomalies, especially cardiac, and the assessment of fetal karyotype. As this is considered a sporadic anomaly, the recurrence risk is low and no genetic origin is known. PMID- 16215331 TI - Impact of controlled-release oxycodone on efficacy beliefs and coping efforts among osteoarthritis patients with moderate to severe pain. AB - OBJECTIVES: This study examines the impact of controlled-release oxycodone (OxyContin) on cognitive-behavioral indicators of efficacy beliefs and coping efforts in patients with osteoarthritis who experience persistent pain. METHODS: This was a double-blind, placebo-controlled study of 104 male and female patients with osteoarthritis who experienced moderate to severe pain. Pain assessment, arthritis helplessness, and coping efforts were assessed before and after for treatment and placebo control groups. RESULTS: Examination of differences between treatment and control groups after 2 weeks revealed significant reductions in reported pain, improvements in coping efficacy, and reductions in helplessness and passive coping in response to controlled-release oxycodone treatment compared to placebo. Changes in pain partially mediated the effects on coping in subsequent assessments. DISCUSSION: The findings indicate that controlled-release oxyco- done treatment accounted for improvements in coping with pain beyond that of placebo controls. This medication may be most beneficial to osteoarthritis patients when incorporated as part of a multidisciplinary approach to pain management. PMID- 16215330 TI - A randomized, controlled trial of manual therapy and specific adjuvant exercise for chronic low back pain. AB - OBJECTIVE: This article examines the effectiveness of manual therapy with specific adjuvant exercise for treating chronic low back pain and disability. METHODS: A single blind, randomized, controlled trial was employed. Patients were prescribed an exercise program that was tailored to treat their musculoskeletal dysfunctions or given a nonspecific program of general stretching and aerobic conditioning. In addition, patients received manual therapy or sham manual therapy. Participants were seen for 6 weekly sessions and were asked to perform their exercise program twice daily. RESULTS: Seventy-two out of 100 patients completed the study. Multivariate tests conducted for measures of pain and disability revealed a significant group by time interaction (P = 0.04 and P = 0.05, respectively), indicating differential change in these measures pretreatment to posttreatment as a function of the treatment received. When controlling for pretreatment scores, patients receiving manual therapy with specific adjuvant exercise reported significant reductions in pain. No change in perceived disability was observed, with the exception that patients receiving sham manual therapy with specific adjuvant exercise reported significantly greater disability at posttreatment. DISCUSSION: Manual therapy with specific adjuvant exercise appears to be beneficial in treating chronic low back pain. Despite changes in pain, perceived function did not improve. It is possible that impacting chronic low back pain alone does not address psychosocial or other factors that may contribute to disability. Further studies are needed to examine the long-term effects of these interventions and to address what adjuncts are beneficial in improving function in this population. PMID- 16215332 TI - Stressful experiences in childhood and chronic back pain in the general population. AB - OBJECTIVES: To determine if stressful experiences in childhood are associated with an increased risk of chronic back problems later in life. METHODS: We conducted a prospective cohort study in the Canadian household population. Study participants were respondents to the first 3 cycles of the National Population Health Survey in Canada who were 18 years of age or older at baseline (n = 9552). Cases of chronic back pain during a 4-year follow-up period were ascertained with an interviewer-administered questionnaire. Stressful experiences in childhood were measured by an index consisting of 7 questions. RESULTS: In multivariate analyses, the risk of back pain was 1.17 (95% confidence interval 0.97-1.41) for 1 stressful event and 1.49 (95% confidence interval 1.21-1.84) for 2 or more events. The effect was consistent across subgroups defined by gender, socioeconomic status, and health status. Specific events associated with an increased risk included fearful experiences, prolonged hospitalization, and parental unemployment. DISCUSSION: Our study shows that persons reporting multiple stressful experiences in childhood are at increased risk of developing chronic back problems. PMID- 16215333 TI - Procedural distress in children with cancer: self-report, behavioral observations, and physiological parameters. AB - OBJECTIVES: To examine the relationship among different indicators of pain and distress, including self-report, behavioral observations, and physiological parameters, in children with cancer undergoing invasive procedures. METHODS: Forty-eight children between the ages of 3.1 and 17.7 years were evaluated while undergoing lumbar punctures. Self-report measures assessed anxiety, pain, self efficacy, expectations of coping strategies, and coping self-efficacy. Parents reported on their own and their child's levels of anxiety, and physicians estimated their own level of stress and technical difficulty in completing the procedure. Behavioral observations were made prior to, during, and after the procedure. Physiological parameters included heart rate, cardiac vagal tone, and salivary cortisol. At the discretion of attending physicians, 32 children received deep sedation, 9 received light sedation, and 7 received cognitive behavioral strategies with topical anesthetic as interventions to manage procedural distress. RESULTS: There was a high degree of consistency within self report, behavioral, and physiological parameters, but correlations between measures in different modalities were low. There were floor effects for most behavioral and self-report measures of distress. Cortisol showed marked changes preprocedure to postprocedure, demonstrating high levels of physiological response despite lack of apparent or perceived discomfort. Heart rate was significantly lower in the group using cognitive-behavioral techniques, especially at the point of needle insertion. DISCUSSION: Self-report measures, behavioral indicators, and physiological changes are not interchangeable outcomes. Treatment strategies were effective for minimizing subjective and behavioral distress, but not necessarily for physiological reactions. Future research should focus on individual differences in these responses, and treatment outcome studies aimed at reducing distress must be clear about the specific goals of intervention. PMID- 16215334 TI - Body movements: an important additional factor in discriminating pain from stress in preterm infants. AB - OBJECTIVES: To describe developmentally appropriate, specific body movements and other biobehavioral responses of preterm infants to a group of routine care giving tasks (Clustered Care), and to compare responses to acute pain with those of Clustered Care. METHODS: In a randomized design, 54 preterm infants were assessed at 32 weeks gestational age during 3 phases of blood collection (Baseline, Lance/squeeze, Recovery) and of diaper changing, measuring abdominal girth and axillary temperature, and mouth care (Baseline, Clustered Care, Recovery) in a neonatal intensive care unit. The Newborn Individualized Developmental Care and Assessment Program and 1 facial action from the Neonatal Facial Coding System, Brow Bulge, were coded from separate continuous bedside video recordings. Heart rate and oxygen saturation were also acquired continuously. RESULTS: Brow Bulge, heart rate, and a subset of 9 Newborn Individualized Developmental Care and Assessment Program movements increased and oxygen saturation decreased significantly to Lance/squeeze compared to Baseline. Similar facial and physiological changes occurred during Clustered Care, but with less intensity. However, infants showed greater frequencies and variety of Newborn Individualized Developmental Care and Assessment Program stress cues during Clustered Care than during Lance/squeeze. Stress cues persisted after Clustered Care, whereas the infants returned to Baseline following Lance/squeeze. DISCUSSION: Changes in facial activity and heart rate remain the most sensitive markers of pain in preterm infants. Tactile procedures, such as diaper changing, produce lower intensity facial and physiological responses than pain procedures, but greater body reactions. Also, the effects from tactile procedures appear to last longer. Adding observations of a small number of specific body movements to the assessment of pain and stress provides complementary information particularly for those infants who may show dampened facial reactivity as a result of repeated pain exposure. PMID- 16215335 TI - "Ow!": spontaneous verbal pain expression among young children during immunization. AB - OBJECTIVES: Although self-reports are a commonly used means of assessing pain in clinical settings, little is understood about the nature of children's spontaneous verbal expressions of pain. The purpose of this study was to describe verbalizations of pain among children receiving a preschool immunization and to examine how pain verbalizations correspond to children's facial expressions and self-reports of pain intensity. METHODS: Fifty-eight children between the ages of 4 years 8 months and 6 years 3 months (67% female) were videotaped while receiving their routine preschool immunization. Global ratings of facial expression and detailed transcription and coding of pain verbalizations were undertaken. Children provided self-reports of pain using a 7-point faces pain scale. RESULTS: Fifty-three percent of children used verbalizations spontaneously to express their pain. The modal verbalization was the interjection "Ow!," which expressed negative affect and was specific to the experience of pain. Older children were less likely to use verbalizations to express their pain. Children who used verbalizations to express pain displayed greater facial reactions to pain and rated their pain experience as being more intense than children who did not use words to express their pain. DISCUSSION: Results indicate that many young children do not spontaneously use verbalizations to express pain from immunization. When 5-year-olds use verbalizations to express pain, the verbalizations are most often brief statements that express negative affect and directly pertain to pain. Knowledge of how children verbalize pain may lead to an improved ability to assess and manage pediatric pain. PMID- 16215336 TI - Oral methadone for chronic noncancer pain: a systematic literature review of reasons for administration, prescription patterns, effectiveness, and side effects. AB - OBJECTIVE: To assess the indications, prescription patterns, effectiveness, and side effects of oral methadone for the treatment of chronic noncancer pain. METHODS: We conducted searches of several electronic databases, textbooks and reference lists for controlled or uncontrolled studies in humans. Effectiveness was assessed using a dichotomous classification of "meaningful" versus "nonmeaningful" outcomes. RESULTS: Twenty-one papers (1 small randomized trial, 13 case reports, and 7 case series) involving 545 patients with multiple noncancer pain conditions were included. In half of the patients, no specific diagnosis was reported. Methadone was administered primarily when previous opioid treatment was ineffective or produced intolerable side effects. Starting dose ranged from 0.2 to 80 mg/day and maximum dose ranged from 20 to 930 mg/day. Pain outcomes were meaningful in 59% of the patients in the uncontrolled studies. The randomized trial demonstrated a statistically significant improvement in pain for methadone (20 mg/day) compared to placebo. Side effects were considered minor. DISCUSSION: Oral methadone is used for various noncancer pain syndromes, at different settings and with no prescription pattern that could be identifiable. Starting, maintenance, and maximum doses showed great variability. The figure of 59% effectiveness of methadone should be interpreted very cautiously, as it seems overrated due to the poor quality of the uncontrolled studies and their tendency to report positive results. The utilization of oral methadone for noncancer pain is based on primarily uncontrolled literature. Well-designed controlled trials may provide more accurate information on the drug's efficiency in pain syndromes and in particular neuropathic pain. PMID- 16215337 TI - Age and the experience of chronic pain: differences in health and quality of life among younger, middle-aged, and older adults. AB - OBJECTIVES: To describe age differences in chronic pain and to evaluate for differences in demographic and health-related variables among younger (18-39 years), middle-aged (40-59 years), and older adults (60-81 years) who reported chronic pain. METHODS: A total of 4000 Norwegian citizens were mailed a questionnaire that measured pain, quality of life, mood, and demographic and health-related variables. RESULTS: Of the total sample (n = 1912), 19.2% of the younger age group, 27.5% of the middle-aged group, and 31.2% of the older group reported chronic pain (ie, >3 months duration). A total of 58.9% of the participants in chronic pain reported having a chronic disease, with the most common being musculoskeletal problems, chronic pain disorder, and osteoarthritis. Participants in the older age group reported pain of longer duration and more comorbidities and received pain treatment more often. They had higher total quality of life scores, were more satisfied with their material comforts and social life, and reported better mood. The middle-aged group reported the largest number of pain locations, reported having fibromyalgia more frequently, and reported that the cause of their pain was not known. They were less satisfied with their social life than the older age group. The younger age group reported the highest rates of injury and accidents as the cause of their pain, and almost 43% of this age group was not receiving any treatment of their chronic pain. CONCLUSION: This study found that the prevalence rates for chronic pain do vary with age and that the middle-aged group may be a high-risk group of patients with chronic pain. PMID- 16215338 TI - Treatment of persistent pain associated with osteoarthritis with controlled release oxycodone tablets in a randomized controlled clinical trial. AB - OBJECTIVE: This study, lasting up to 90 days, was undertaken in patients with osteoarthritis with persistent moderate to severe pain uncontrolled by standard therapy (nonsteroidal anti-inflammatory drugs, acetaminophen, and/or short-acting opioids) to evaluate functional outcomes, as well as efficacy and safety, of controlled-release oxycodone versus placebo. METHODS: One hundred seven patients received either controlled-release oxycodone or placebo every 12 hours in this double blind, randomized, placebo-controlled, parallel-group study. Stable previous regimens of acetaminophen or nonsteroidal anti-inflammatory agents were allowed to continue. Primary efficacy variables included Brief Pain Inventory average pain intensity scores at completion of initial titration, Western Ontario and McMaster Universities Osteoarthritis Index scores at days 30 and 60, and the percentage of patients discontinuing due to inadequate pain control. RESULTS: Controlled-release oxycodone was significantly superior to placebo in decreasing average pain intensity and in reducing pain-induced interference with general activity, walking ability (except at day 30), and normal work, as well as mood, sleep, relations with people (at days 60 and 90), and enjoyment in life. Daily functioning, as measured by the Western Ontario and McMaster Universities Osteoarthritis Index, was also significantly improved in the controlled-release oxycodone group. In the placebo group, a significantly greater percentage of patients discontinued due to inadequate pain control. Adverse events were consistent with opioid adverse events, and no safety concerns were noted. DISCUSSION: Treatment with controlled-release oxycodone of patients with osteoarthritis with persistent moderate to severe pain uncontrolled by standard therapy resulted in significant pain control and improvements in physical functioning. PMID- 16215340 TI - Consistent management of repeated procedural pain with sucrose in preterm neonates: Is it effective and safe for repeated use over time? AB - OBJECTIVE: Preterm neonates undergo numerous painful procedures in the neonatal intensive care unit (NICU). Sucrose, with and without pacifiers, is effective and safe for relieving pain from single painful events. However, repeated use of sucrose for multiple painful procedures has not been adequately evaluated. The study objectives were to: 1) determine the efficacy and safety of consistent management of repeated procedural pain with sucrose; and 2) explore the impact of consistent pain management on clinical outcomes and neurobiological risk status. METHODS: Sixty-six preterm infants were randomized to receive standard care (positioning and swaddling; n = 21), sterile water plus pacifier (n = 23), or 24% sucrose plus pacifier (n = 22) prior to all painful procedures in the NICU during the first 28 days of life. Also, at a routine heel lance at 7, 14, 21, and 28 days of life, data were collected to determine efficacy and immediate and long term adverse events. Data on clinical outcomes and neurobiological risk status were collected at 28 days or NICU discharge. RESULTS: There was no intervention by time interaction (P = 0.60). There was a significant main effect of intervention (P = 0.03) between the sucrose plus pacifier group, and the standard care group (P = 0.01), but there was no main effect of time (P = 0.72). No group differences existed for adverse events, clinical outcomes, or neurobiological risk status. DISCUSSION: Consistent management of painful procedures with sucrose plus pacifier was effective and safe for preterm neonates during their stay in the NICU. Further exploration of consistent pain management with sucrose on clinical, developmental, and neurobiological outcomes is required. PMID- 16215339 TI - The efficacy of preoperative versus postoperative rofecoxib for preventing acute postoperative dental pain: a prospective randomized crossover study using bilateral symmetrical oral surgery. AB - BACKGROUND: Previous data have demonstrated that rofecoxib has good analgesic efficacy for acute postoperative dental pain. However, up to half of these patients require rescue analgesics within the first 24 hours. As the timing of analgesic interventions may be an important factor in pain control, the present study tested the hypothesis that rofecoxib administered preoperatively would improve the analgesic efficacy and reduce rescue analgesic requirements within the first 24 hours compared with postoperative administration. METHODS: This was a double-blind, randomized, crossover study where 45 patients had each of their identical impacted mandibular third molars removed under local anesthesia on 2 separate occasions. Patients acted as their own control; one side was pretreated with rofecoxib 50 mg, 2 hours before surgery, followed by placebo 15 minutes after surgery, and the contralateral side was pretreated with placebo 2 hours before surgery and posttreated with rofecoxib 50 mg 15 minutes after surgery. The difference in postoperative pain between 2 sides was assessed by 4 primary end points: pain intensity as measured by a 100-mm visual analogue scale hourly for 12 hours, time to rescue analgesic, postoperative analgesic consumption, and patient's global assessment. RESULTS: Patients reported significantly lower pain scores (P = 0.04), longer time to rescue analgesic (P = 0.02), lesser postoperative analgesic consumption (P = 0.008), and better global assessment (P = 0.01) in the pretreated compared with the posttreated sides. There were significantly more patients in the pretreated group who did not required rescue analgesic within the first 24 hours (80% vs. 58%, P = 0.01), and the pain scores were extremely low in both groups during the 12 hours postoperative period (9.8 +/- 5.0 mm vs. 14.3 +/- 7.4 mm). CONCLUSION: Rofecoxib is an excellent analgesic for preventing postoperative dental pain and when given 2 hours preoperatively rendered most patients relatively pain free, requiring no rescue analgesics on the first postoperative day. PMID- 16215341 TI - Effectiveness of pamidronate for treating intractable chronic neuropathic pain: case report of two adolescents. AB - The objective of this study is to evaluate the effectiveness of pamidronate for the treatment of chronic neuropathic pain refractory to previous management. Intravenous pamidronate (60 mg/day for 3 days) was administered to 2 adolescents with neuropathic pain refractory to previous multidisciplinary treatments. Pain intensity, functional improvement, and adverse effects were evaluated. There were no significant reductions in pain intensity or improvements in function initially or at follow-up at 1 week, 1 month and 4 months. Minor side effects included myalgia and skeletal pain that were relieved by acetaminophen. There was no long term morbidity. In contrast to recent positive reports in adults, pamidronate was not effective in decreasing pain or improving function in 2 adolescents with chronic neuropathic pain. Pamidronate may be effective only when pain is accompanied by abnormal bone density. Future trials should include scans to document bone density pretherapy and posttherapy. PMID- 16215342 TI - Computed tomography-guided anterior approach to the superior hypogastric plexus for noncancer pelvic pain: a report of two cases. AB - The objective of this study was to evaluate the possibility and describe the methodology of a computed tomography-guided anterior approach to superior hypogastric plexus block for noncancer pain. A computed tomography-guided anterior approach to the superior hypogastric plexus was used in 2 patients with pelvic pain and anatomic disturbance of the lumbar spine, which was a contraindication to the conventional dorsal approach. The first case was a 43 year-old patient suffering from burning pain of the urethra. Pain relief using analgesics and antidepressants was insufficient. The posterior approach was excluded due to coexisting irritation of the L5 nerve root. The second case was a 68-year-old man suffering from chronic burning and itching pain of the urethra and glans penis. Conservative therapy (anti-inflammatory drugs, tramadol, spasmolytics) failed to provide satisfactory pain relief. The posterior approach was contraindicated because of laterally prominent L5 vertebral body osteophytes. Both patients received a prognostic block to the superior hypogastric plexus via the anterior approach guided by computed tomography. Visual analog scale scores prior to the block were 6 to 7 and 5 to 6, respectively. The visual analog scale scores 24 hours after the block were 1 and 0, respectively. The second patient received a permanent neurolytic block via the anterior approach to provide long term pain relief. In conclusion, the authors describe the computed tomography guided anterior approach to the superior hypogastric plexus for chronic pelvic pain. The technique is simple to perform, and the analgesic effect is satisfactory. More extensive studies are necessary to evaluate the safety of this approach. PMID- 16215344 TI - Critical illness in pregnancy: an overview. AB - OBJECTIVES: To provide an overview of the critical illnesses that afflict the pregnant patient. DESIGN: A comprehensive review of the literature was performed with Pubmed Medline using medical subject headings of pregnancy, critical care, epidemiology, prognostic score, dyspnea, fever, and jaundice. RESULTS: Although pregnant women constitute a small number of admissions to an intensive care unit, they pose a challenge to the health care team. The physiological changes that occur in pregnancy stress and limit the compensatory response required to adapt to a superimposed acute illness. Although a number of conditions are uniquely associated with pregnancy, other medical conditions may complicate the course of pregnancy. The pregnant state also increases susceptibility to a number of illnesses. In this overview, we reviewed the epidemiology and different clinical presentations of critical illness in a pregnant patient. CONCLUSIONS: The critically ill pregnant patient presents a challenge to the critical care physician due to unique physiology and specific medical disorders in this population. PMID- 16215345 TI - Physiologic changes in pregnancy and their impact on critical care. AB - OBJECTIVES: To describe the major pregnancy-induced physiologic changes that affect the care of patients who are critically ill. METHODS: We reviewed relevant textbooks and articles pertaining to physiologic alterations produced by pregnancy. Those changes that have a bearing on patients admitted to an intensive care unit were abstracted, summarized, and organized by organ system. CONCLUSIONS: Several organ systems manifest significant change in function during pregnancy. The cardiovascular and pulmonary systems are the two most important with regard to critical care. PMID- 16215346 TI - Airway problems in pregnancy. AB - OBJECTIVES: To provide a current review of the literature regarding airway problems in pregnancy and management. BACKGROUND: Obstetrical anesthesia is considered to be a high-risk practice that exposes the anesthesiologist to increased medicolegal liability. Anesthetic management of a parturient is a challenge because it involves simultaneous care of both mother and baby. Failure to appropriately manage a difficult or failed intubation increases the risk of hypoxemic cardiopulmonary arrest and/or pulmonary aspiration, resulting in a high probability of maternal morbidity and mortality. DATA: Anesthesia is the seventh leading cause of maternal mortality in the United States. Anatomic and physiologic changes during pregnancy place the parturient at increased risk for airway management problems. It is essential to perform a thorough preanesthetic evaluation and identify the factors predictive of difficult intubation. Airway devices such as the laryngeal mask airway, ProSeal, intubating laryngeal mask airway, Combitube, and laryngeal tube are described and have been used during failed intubation in pregnant patients. CONCLUSION: Teamwork between an anesthesiologist and an obstetrician is absolutely essential for the safety of both the mother and baby. Most of us tend to agree that airway emergencies have a way of occurring at the worst possible times. It is essential that all anesthesia care practitioners must have a preconceived and well thought-out algorithm and emergency airway equipment to deal with airway emergencies during difficult or failed intubation of a parturient. PMID- 16215347 TI - Acute respiratory distress syndrome in pregnancy. AB - OBJECTIVE: To summarize the pathophysiology and treatment of acute lung injury and acute respiratory distress syndrome (ARDS) during pregnancy. DATA SOURCE: Review of select articles from MEDLINE, including published abstracts, case reports, observational studies, controlled trials, review articles, and institutional experience. DATA SUMMARY: ARDS occurs in pregnancy and may have unique causes. Despite extensive clinical research to improve the management of ARDS, mortality remains high, and few strategies have shown a mortality benefit. Furthermore, in most published studies, pregnancy is an exclusionary criterion, and thus, few treatments have been adequately evaluated in obstetric populations. The treatment of ARDS in pregnancy is extrapolated from studies performed in the general ARDS patient population, with consideration given to the normal physiologic changes of pregnancy. In general, the best support of the fetus is support of the mother. From the age of viability (24-26 wks at most institutions) until full term, decisions regarding delivery should be made based primarily on the standard obstetric indications. CONCLUSIONS: Little evidence exists regarding the management of ARDS specifically in pregnancy, and thus, treatment approaches must be drawn from studies performed in a general patient population. A multidisciplinary approach involving maternal-fetal medicine, neonatology, anesthesiology, and intensivist clinicians is essential to optimizing maternal and fetal outcomes. PMID- 16215348 TI - Amniotic fluid embolism. AB - INCIDENCE: Amniotic fluid embolism is a catastrophic syndrome that occurs during pregnancy or in the immediate postpartum period. Multiple case reports have described the clinical findings and have reported variable success with supportive care. There has been discrepancy with respect to the incidence and mortality of amniotic fluid embolism. One likely explanation for this inconsistency is the lack of sensitive and specific diagnostic studies to definitively identify cases of amniotic fluid embolism, leading to both over- and underreporting. Despite the variation in reported incidence and mortality, amniotic fluid embolism remains a life-threatening condition with significant morbidity and mortality for the pregnant woman. It is the fifth most common cause of maternal mortality in the world. DIAGNOSIS: The diagnosis of amniotic fluid embolism continues to be a clinical diagnosis and a diagnosis of exclusion based on the rapid development of a complex constellation of findings with sudden cardiovascular collapse, acute left ventricular failure with pulmonary edema, disseminated intravascular coagulation, and neurologic impairment. Given the significant morbidity and mortality associated with this condition, a high index of suspicion is warranted. Suspected risk factors have included tumultuous labor, trauma, multiparity, increased gestational age, and increased maternal age. However, many patients who develop amniotic fluid embolism have no obvious risk factors. MANAGEMENT: Patients with amniotic fluid embolus are best managed using a multidisciplinary approach. There are no pharmacologic or other therapies that prevent or treat the amniotic fluid embolism syndrome, and supportive care typically involves aggressive treatment of multiple types of shock simultaneously. In this article we discuss the clinical presentation of amniotic fluid embolism syndrome as well as current opinions regarding pathophysiology, diagnosis, and management. PMID- 16215349 TI - Sepsis during pregnancy. AB - OBJECTIVES: To provide a current review of the literature regarding the assessment and management of sepsis during pregnancy. DESIGN: A comprehensive review of current English-language literature search was performed with Ovid MEDLINE using the Medical Subject Headings pregnancy and sepsis, with Medical Subject Headings or keywords seeking randomized controlled trials and clinical reports, and by reviewing the bibliographies of clinical practice guidelines. RESULTS: Sepsis-related maternal morbidity and mortality is a significant and persistent problem in the modern critical care obstetric unit. The management of sepsis during pregnancy is challenging. The obstetric intensivist must simultaneously discern the effect of maternal physiologic changes on fetal vulnerability and the effect of the fetus on maternal status throughout the various phases of pregnancy. Little direct evidence exists to validate the extrapolation of some sepsis treatment modalities from other nonpregnant patient populations. Nevertheless, early detection, accurate diagnosis, and aggressive appropriate treatment strategies may significantly improve outcome. Approaches like the Surviving Sepsis Campaign guidelines are unproven but seem reasonable and practical. CONCLUSIONS: Sepsis during pregnancy is uncommon yet potentially fatal. Diagnostic and therapeutic guidelines should predominantly pattern those currently utilized for nonpregnant patients. PMID- 16215350 TI - Pulmonary embolism during and after pregnancy. AB - BACKGROUND: Venous thromboembolic disease is among the most common causes of morbidity and mortality during pregnancy. The clinical evaluation alone is insufficient for the diagnosis of venous thromboembolic disease, and the normal pregnant state makes this evaluation even more challenging. DIAGNOSIS: Objective testing is the mainstay of diagnosis, including compression ultrasound, impedance plethysmography, ventilation-perfusion scanning, computed tomography scanning, and pulmonary angiography. All of these tests can be safety performed during pregnancy. TREATMENT: If deep vein thrombosis or pulmonary embolism is diagnosed, anticoagulation should be initiated. Either (unfractionated) heparin or low molecular weight heparin is an acceptable treatment for acute venous thromboembolic disease. Both have risks and benefits, but both can be used safely during pregnancy. Intravenous heparin is the treatment of choice surrounding delivery due to its short half life. Because of the risk of adverse effects on the fetus, warfarin is not generally used during pregnancy. Unstable pulmonary embolism is difficult to treat during pregnancy, as there are minimal data regarding the safety and efficacy of thrombolytic therapy, inferior vena cava filters, and embolectomy during pregnancy. Case reports and case series suggest that thrombolytic therapy may be associated with lower risks of fetal loss than embolectomy. CONCLUSIONS: Venous thromboembolic disease is a significant cause of morbidity and mortality during pregnancy and the puerperal period. Objective testing is critical to establish the diagnosis and can be safely performed during pregnancy. Anticoagulation with heparin is the mainstay of therapy during the pregnancy, but patients may be transitioned to warfarin after delivery. PMID- 16215351 TI - Ovarian hyperstimulation syndrome. AB - OBJECTIVE: Ovarian hyperstimulation syndrome (OHSS) is a rare iatrogenic complication of ovarian stimulation usually occurring during the luteal phase or during the early part of pregnancy. OHSS is a potential complication of ovarian induction by almost every agent used for ovarian stimulation. Today, due to aggressive treatment protocols including the development of in vitro fertilization and cryopreservation with the goal of obtaining sufficient numbers of oocytes and embryos, an increased risk of developing OHSS is present. OHSS is now becoming increasingly more recognized due to the higher number of women undergoing assisted reproductive techniques. DESIGN: Review of the literature regarding ovarian hyperstimulation syndrome. METHODS: A review of the epidemiology, pathophysiology, risk factors, classification, clinical features, and treatment and prevention of OHSS. CONCLUSION: OHSS can be thought of as the loss of control over the hyperstimulation of the ovaries. Although the prevalence of the severe form of OHSS is small, it is important to remember that OHSS is usually an iatrogenic complication of a nonvital treatment that has the potential for a fatal outcome. Therefore, critical care physicians play an integral part in the care of these patients and therefore should be familiar with and recognize the various clinical manifestations and potential outcomes of this entity. PMID- 16215352 TI - Acute hypertensive emergencies in pregnancy. AB - OBJECTIVE: Obstetrical hypertensive emergencies are life-threatening conditions involving significant risk to both the mother and fetus. Aggressive treatment of the maternal hypertensive state requires an initial consideration of the effect of treatment on the fetus, via changes to the uteroplacental circulation with treatment. The challenge then is to correct blood pressure using appropriate, safe pharmacologic agents to prevent catastrophic maternal consequences, while minimizing acute changes to placental perfusion and any corresponding fetal ill effects. Hypertension in pregnancy may be one manifestation of a multiple-system pathologic process, as is the case in preeclampsia. Blood pressure control, along with delivery, will be the first step in treating the renal, hematologic, hepatic, and cardiac dysfunction that can be seen in preeclampsia. DESIGN: A review of medications most commonly used for hypertensive emergencies in pregnancy. CONCLUSIONS: Hypertensive emergencies in pregnancy require prompt evaluation and treatment in an intensive care setting to prevent untoward effects to both the fetus and mother. PMID- 16215354 TI - Acute asthma in pregnancy. AB - OBJECTIVES: To provide an overview on the interrelationship between asthma and pregnancy, focusing on management of pregnant women presenting with an acute severe exacerbation. DESIGN: A review of the current English-language published clinical trials was performed based on MEDLINE search using the Medical Subject Headings pregnancy and asthma. Current reviews on the topic and practice guidelines were also reviewed. RESULTS: Asthma is the most common medical condition to complicate pregnancy, and episodes of acute asthma requiring emergency department visits or hospitalization have been reported in 9-11% of pregnant women managed by asthma specialists. Pregnancy can affect the course of asthma, and the risk of asthma exacerbations requiring intervention in pregnant women is higher than in nonpregnant women. Similarly, asthma can affect pregnancy outcomes. Maternal inflammatory pathways may contribute to the poor pregnancy outcomes, especially in women with uncontrolled asthma. Although data on the effects of maternal asthma on pregnancy have been conflicting, mainly because many published studies have not corrected for asthma severity, it has generally been observed that poorly managed asthma during pregnancy is associated with a higher risk of preterm delivery, low birth weight, and complications such as preeclampsia. Optimal therapy of asthma has been shown to contribute to improved maternal and fetal outcomes. CONCLUSIONS: Asthma can complicate the course of pregnancy, and pregnancy can worsen asthma control in some women. Optimal management of asthma during pregnancy is key in ensuring the safety of the mother and the fetus. PMID- 16215353 TI - Exacerbation of underlying pulmonary disease in pregnancy. AB - OBJECTIVE: The presence of underlying pulmonary disease in women of childbearing potential can present a significant challenge during pregnancy and the postpartum period. Management of the underlying disease, recognizing and preventing disease progression, and, most important, managing and minimizing toxic side effects of various therapies require the expertise of an interdisciplinary team. This team must involve close collaboration between intensive care physicians, pulmonary physicians, and high-risk obstetricians familiar with these disease states in an effort to minimize fetal and maternal morbidity and mortality. We will review the impact of the pregnant state in lung transplant recipients, patients with pulmonary arterial hypertension, and patients with underlying cystic fibrosis. DESIGN: Review of the literature in regards to pregnancy outcomes and issues for patients with cystic fibrosis, pulmonary hypertension, and lung transplants. METHODS: A review of the epidemiology, pathophysiology, risk factors, classification, clinical features, and outcomes for pregnant patients with underlying pulmonary diseases. CONCLUSIONS: Safety of pregnancy in the female lung transplant recipient concerns three outcomes: maternal outcome, fetal outcome, and transplanted graft outcome. PMID- 16215355 TI - Cardiac arrest in pregnancy and somatic support after brain death. AB - OBJECTIVE: To review the important causes of cardiopulmonary arrest during pregnancy and the recommended modifications to resuscitation protocols when applied to pregnant patients, including the indications for perimortem cesarean section and the expected fetal outcomes, and to review the literature regarding extended somatic support after brain death during pregnancy. DATA SOURCES: MEDLINE review of publications relating to cardiac arrest and resuscitation in pregnancy, physiologic changes after brain death, and attempted somatic support of brain-dead pregnant women. CONCLUSIONS: Cardiac arrest during pregnancy is rare, but it is important to recognize the causes, which may be either unrelated to pregnancy or unique to the pregnant woman. For the most part, the resuscitation protocol is the same as for nonpregnant victims of cardiac arrest, with a few important modifications, including especially the need for relieving aortocaval compression by the gravid uterus, the need for rapid intubation, and the importance of rapid perimortem cesarean delivery when indicated. In those rare cases of brain death occurring in a pregnant patient, continued somatic support of the mother may be possible, even for prolonged periods, to extend the pregnancy and further fetal maturation. The expected physiologic changes after brain death, challenges to successful somatic support, and specific recommendations regarding organ support of the brain-dead pregnant woman are reviewed. PMID- 16215356 TI - Hepatic disease and pregnancy: an overview of diagnosis and management. AB - OBJECTIVES: To provide a review of the current literature on hepatic disease in pregnancy, outlining the incidence, pathophysiology, diagnosis, and management of major diseases in this category. DESIGN: A thorough review of expert analysis, case reports, and randomized clinical trials was used to assess current methods of managing the major diseases related to hepatic dysfunction in pregnancy. A review of bibliographies was also utilized. RESULTS: Hepatic disease complicates nearly 3% of all pregnancies and is a significant cause of morbidity during the gravid state. However, several diseases, including HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets), acute fatty liver of pregnancy, acute cholestasis of pregnancy, ruptured liver hematoma, and preeclampsia, can be managed with successful outcomes for both the mother and fetus if diagnosed in early stages. Astute clinical judgment and laboratory evaluation of the patient is vital in the appropriate diagnosis of hepatic disease in pregnancy. CONCLUSIONS: Early intervention and appropriate diagnosis can substantially reduce the morbidity and mortality associated with hepatic derangements of pregnancy. PMID- 16215357 TI - Peripartum cardiomyopathy. AB - OBJECTIVE: To provide a review of the cardiac and obstetrical literature regarding the development of peripartum cardiomyopathy and, in particular, to examine risk factors, incidence, diagnosis, prognosis, and evidence-based treatment modalities. DESIGN: An extensive review of the current literature. RESULTS: Peripartum cardiomyopathy is a cardiomyopathy of unknown cause that occurs in pregnant females, most commonly in the early postpartum period. It shares many clinical characteristics with idiopathic dilated cardiomyopathy but occurs at a younger age and is associated with a better prognosis. Diagnosis is based upon the clinical presentation of congestive heart failure and objective evidence of left ventricular systolic dysfunction. Conventional pharmacologic therapy for congestive heart failure, such as diuretics, digoxin, angiotensin converting enzyme inhibitors, angiotensin-receptor blockers, and beta-adrenergic blockers, are routinely used and are quite effective. For those patients who remain refractory to conventional pharmacologic therapy, cardiac transplantation and mechanical circulatory support are viable options. CONCLUSION: Mortality rates in peripartum cardiomyopathy have decreased, and this is most likely related to advances over the past 5 yrs in medical therapy for heart failure. Aggressive use of implantable defibrillators has significantly reduced the risk of sudden death in these patients. For >50% of peripartum cardiomyopathy patients, left ventricular function normalizes with pharmacologic therapy. However, subsequent pregnancies almost always are associated with recurrence of left ventricular systolic dysfunction. PMID- 16215358 TI - Diabetic ketoacidosis in pregnancy. AB - OBJECTIVE: The development of diabetic ketoacidosis in pregnancy is a medical emergency, requiring treatment in an intensive care setting. Both the mother and the fetus are at risk for significant morbidity and mortality. Physiologic changes unique to pregnancy provide a background for the development of diabetic ketoacidosis. An understanding of these physiologic changes assists in the management of the two patients being treated. Treatment of the patient with diabetic ketoacidosis includes insulin therapy and careful fluid management; recommendations for management are presented. PATIENTS: Pregnant women, either with preexisting diabetes or with diabetes diagnosed during pregnancy. CONCLUSIONS: Prompt recognition of the clinical manifestations of diabetic ketoacidosis, followed by appropriate, timely treatment will optimize outcome for the pregnant woman and her fetus. PMID- 16215359 TI - Hemodynamic assessment in a pregnant and peripartum patient. AB - PATHOPHYSIOLOGY: Critical care in obstetrics has many similarities in pathophysiology to the care of nonpregnant women. However, changes in the physiology of pregnant woman necessary to maintain homeostasis for both mother and fetus, especially during critical illness, result in complex pathophysiology. Understanding the normal physiologic changes during pregnancy, intrapartum, and postpartum is the key to managing critically ill obstetric patients with underlying medical diseases and pregnancy-related complications. HEMODYNAMIC MONITORING: When the pathophysiology of critically ill obstetric patients cannot be explained by noninvasive hemodynamic monitoring and the patient fails to respond to conservative medical management, invasive hemodynamic monitoring may be helpful in guiding management. Most important, the proper interpretation of hemodynamic data is predicated on knowledge of normal values during pregnancy and immediately postpartum. Invasive hemodynamic monitoring with pulmonary artery catherization has been used in the obstetric population, particularly in patients with severe preeclampsia associated with pulmonary edema and renal failure. PMID- 16215360 TI - Neurologic disorders in pregnancy. AB - BACKGROUND: Neurologic dysfunction, coma, and seizures are common in obstetric patients in the intensive care unit. OBJECTIVE: To review common neurologic disorders resulting in critical illness in pregnancy. REVIEW: Obstetric disorders causing coma and seizures include eclampsia, acute fatty liver of pregnancy, and amniotic fluid embolism. Preexisting disorders such as epilepsy may worsen in one third of pregnant patients, and seizures are common during labor. Changes in hemodynamics, blood volume, and hormonal effects on the vessel wall increase risk of bleeding from berry aneurysms and arteriovenous malformations during pregnancy and the postpartum period. Acute intermittent porphyria produces seizures and hypertension, closely mimicking eclampsia. Cerebral venous sinus thrombosis is common in postpartum patients, especially in developing countries. Brain tumors invariably enlarge during pregnancy because of fluid retention and the presence of estrogen and progesterone receptors on tumor cells. Infections such as cerebral malaria and acute viral hepatitis with fulminant hepatic failure are common causes of coma and seizures during pregnancy in tropical regions of Asia, Africa, and Latin America. Patients may be admitted to the intensive care unit with type II respiratory failure due to myasthenic crisis, Guillain-Barre syndrome and spinal cord disease. Relapses of multiple sclerosis are infrequent during pregnancy but increase in the postpartum period. CONCLUSIONS: In all instances, the effects of the disorders, diagnostic tests, and treatment on the fetus must be carefully weighed. Prompt delivery may be lifesaving for mother and fetus in conditions such as eclampsia and acute fatty liver of pregnancy; expectant treatment may be more appropriate in others. PMID- 16215361 TI - Acute renal failure in pregnancy. AB - OBJECTIVES: To provide an evidence-based, up-to-date review of the literature regarding the assessment and management of acute renal failure that may affect women during pregnancy and the postpartum period. DESIGN: A review of the current literature was performed. RESULTS: Acute renal failure is a rare complication of pregnancy but is associated with significant morbidity and mortality. Management requires knowledge of the renal physiologic changes occurring in pregnancy and the relevant diagnoses, both pregnancy-specific and those that may coincidentally occur with pregnancy. In addition, fetal effects must be taken into consideration. CONCLUSIONS: Ideal care for women with acute renal failure in pregnancy or postpartum requires a multidisciplinary approach that may include maternal-fetal medicine, critical care medicine, nephrology, and neonatology specialists. PMID- 16215362 TI - Trauma in pregnancy. AB - OBJECTIVE: The objective of this article was to review the existing standards of practice regarding trauma which occurs during pregnancy. DESIGN: The design of this study was to review the available data from the surgical and obstetrical literature regarding trauma during pregnancy. The design was also to incorporate the contemporary recommendations from the trauma resuscitation courses relating to trauma during pregnancy. RESULTS: Trauma occurs in 5% of pregnancies. A fetus is not considered to be viable until week 25. Motor vehicle accidents account for more than 50% of all trauma during pregnancy, with 82% of fetal deaths occurring during these automobile accidents. With life threatening trauma a 50% fetal loss rate exists. As anatomy, physiology, and even laboratory findings change during pregnancy, the clinician must consider both patients, the mother and fetus. Following blunt trauma abruption of the placenta is the more common cause of fetus loss. Anterior abdominal penetrating trauma almost never fails to injury the uterus and fetus in the last half of pregnancy. Preventive strategies exist in the areas of social violence, automobile restraints and use of alcohol and drugs by the mother. Perimortem caesarian section is rarely successful. CONCLUSIONS: Trauma during pregnancy is uncommon, but with increasing trauma severity leads to increased fetal loss. Preventive strategies exist and when admitted monitoring standards should be followed. PMID- 16215363 TI - Pneumonia in pregnancy. AB - OBJECTIVE: Historically, pneumonia during pregnancy has been associated with increased morbidity and mortality compared with nonpregnant women. The goal of this article is to review current literature describing pneumonia in pregnancy. This review will identify maternal risk factors, potential complications, and prenatal outcomes associated with pneumonia and describe the contemporary management of the varied causes of pneumonia in pregnancy. RESULTS: Coexisting maternal disease, including asthma and anemia, increase the risk of contracting pneumonia in pregnancy. Neonatal effects of pneumonia in pregnancy include low birth weight and increased risk of preterm birth, and serious maternal complications include respiratory failure. Community-acquired pneumonia is the most common form of pneumonia in pregnancy, with Streptococcus pneumoniae, Haemophilus influenzae, and Mycoplasma pneumoniae accounting for most identified bacterial organisms. Beta-lactam and macrolide antibiotics are considered safe in pregnancy and are effective for most community-acquired pneumonia in pregnancy. Viral respiratory infections, including varicella, influenza, and severe acute respiratory syndrome, can be associated with maternal pneumonia. Current antiviral and respiratory therapies can reduce maternal morbidity and mortality from viral pneumonia. Influenza vaccination can reduce the prevalence of respiratory hospitalizations among pregnant women during influenza season. Pneumocystis pneumonia continues to carry significant maternal risk to an immunocompromised population. Prevention and treatment of Pneumocystis pneumonia with trimethoprim/sulfamethoxazole is effective in reducing this risk. CONCLUSIONS: Prompt diagnosis and treatment with contemporary antimicrobial therapy and intensive care unit management of respiratory compromise has reduced the maternal morbidity and mortality due to pneumonia in pregnancy. Prevention with vaccination in at-risk populations may reduce the prevalence and severity of pneumonia in pregnant women. PMID- 16215365 TI - Comparison of prone positioning and high-frequency oscillatory ventilation in patients with acute respiratory distress syndrome. AB - OBJECTIVE: Both prone position and high-frequency oscillatory ventilation (HFOV) have the potential to facilitate lung recruitment, and their combined use could thus be synergetic on gas exchange. Keeping the lung open could also potentially be lung protective. The aim of this study was to compare physiologic and proinflammatory effects of HFOV, prone positioning, or their combination in severe acute respiratory distress syndrome (ARDS). DESIGN: : Prospective, comparative randomized study. SETTING: A medical intensive care unit. PATIENTS: Thirty-nine ARDS patients with a Pao2/Fio2 ratio <150 mm Hg at positive end expiratory pressure > or =5 cm H2O. INTERVENTIONS: After 12 hrs on conventional lung-protective mechanical ventilation (tidal volume 6 mL/kg of ideal body weight, plateau pressure not exceeding the upper inflection point, and a maximum of 35 cm H2O; supine-CV), 39 patients were randomized to receive one of the following 12-hr periods: conventional lung-protective mechanical ventilation in prone position (prone-CV), HFOV in supine position (supine-HFOV), or HFOV in prone position (prone-HFOV). MEASUREMENTS AND MAIN RESULTS: Prone-CV (from 138 +/ 58 mm Hg to 217 +/- 110 mm Hg, p < .0001) and prone-HFOV (from 126 +/- 40 mm Hg to 227 +/- 64 mm Hg, p < 0.0001) improved the Pao2/Fio2 ratio whereas supine-HFOV did not alter the Pao2/Fio2 ratio (from 134 +/- 57 mm Hg to 138 +/- 48 mm Hg). The oxygenation index ({mean airway pressure x Fio2 x 100}/Pao2) decreased in the prone-CV and prone-HFOV groups and was lower than in the supine-HFOV group. Interleukin-8 increased significantly in the bronchoalveolar lavage fluid (BALF) in supine-HFOV and prone-HFOV groups compared with prone-CV and supine-CV. Neutrophil counts were higher in the supine-HFOV group than in the prone-CV group. CONCLUSIONS: Although HFOV in the supine position does not improve oxygenation or lung inflammation, the prone position increases oxygenation and reduces lung inflammation in ARDS patients. Prone-HFOV produced similar improvement in oxygenation like prone-CV but was associated with higher BALF indexes of inflammation. In contrast, supine-HFOV did not improve gas exchange and was associated with enhanced lung inflammation. PMID- 16215366 TI - Cardiovascular response to dopamine and early prediction of outcome in septic shock: a prospective multiple-center study. AB - OBJECTIVES: To compare mortality rates between dopamine-sensitive (Dopa-S) and dopamine-resistant (Dopa-R) septic shock patients, the latter group defined by a mean arterial pressure <70 mm Hg despite the use of 20 mug/kg/min dopamine. DESIGN: A human, prospective observational, multiple-center, clinical trial. SETTING: Ten intensive care units from ten hospitals. PATIENTS: 110 patients with septic shocks. INTERVENTIONS: Following volume resuscitation, patients were treated by a rapid increase in dopamine infusion from 10 to 20 mug/kg/min. If mean arterial pressure remained <70 mm Hg, dopamine treatment was promptly switched to norepinephrine or epinephrine. MEASUREMENTS AND MAIN RESULTS: Dopamine sensitivity, arterial gas, lactate, and organ system failure scores were measured at admission and after 6, 12, 24, 48, 72, 96, and 120 hrs. The overall 28-day mortality rate was 54% for the entire population under study. In multivariate analysis, independent predictors of death were dopamine resistance (odds ratio, 9.5; 95% confidence interval, 3-25), arterial lactate >3.5 mmol/L (odds ratio, 1.75; 95% confidence interval, 1.06-2.55), and Sepsis-related Organ Failure Assessment score >10 (odds ratio, 1.40; 95% confidence interval, 1.07 2.12). Of the 110 patients studied, 66 were observed to be resistant to dopamine (60%). In the Dopa-S group, the 28-day mortality rate was 16% (seven of 44 patients) compared with 78% (52 of 66 patients) in the Dopa-R group (p = .0006). The capacity of dopamine resistance to predict death was associated with a sensitivity of 84% and a specificity of 74%. At 24 hrs, the association of dopamine resistance to a lactate level >3.5 mmol/L improved the prognostic value (sensitivity, 90%, specificity, 92%). CONCLUSIONS: Dopamine sensitivity is associated with decreased mortality rate. Early recognition of dopamine resistant septic shock could allow for better screening of patients with an ominous prognosis. PMID- 16215367 TI - Influence of entry criteria on mortality risk and number of eligible patients in recent studies on severe sepsis. AB - OBJECTIVE: To understand the impact of patient selection criteria used in recent sepsis trials on baseline mortality risk and number of eligible patients. DESIGN: Observational cohort study, with retrospective analysis of prospectively collected data. METHODS AND MAIN RESULTS: Using a MEDLINE search, we selected recent randomized controlled trials in patients with severe sepsis and studied the mortality rate in the control groups of these trials. Nine articles fulfilled the search criteria and were used in our analyses. The 28-day mortality rate in the control groups of these trials varied between 28.0% and 89.0%. Differences in this mortality rate might be due to the use of different entry criteria but also to other factors that vary between the trials. To eliminate the influence of these confounding factors when studying the effect of the use of entry criteria on baseline mortality risk and number of eligible patients, we projected the entry criteria of these nine trials on a large independent database of >70,000 Dutch intensive care patients admitted between 1996 and 2003. This yielded nine groups of patients who would have been eligible for the respective trials. The percentage of patients who would have been eligible for these trials varied between 1.5% and 6.0%. Six of these groups had a similar intensive care mortality rate (between 25.0% and 28.9%). The projection of the entry criteria of the three other trials onto the database resulted in groups of patients with considerably higher intensive care mortality. For in-hospital mortality rate in these groups, similar results were found. CONCLUSIONS: The majority of the trials we studied used entry criteria that select patients with a similar mortality risk. This suggests that differences in baseline mortality risk reported in recent sepsis trials are to be attributed to other factors that vary between trials rather than to differences in entry criteria. However, entry criteria do have an important influence on the number of eligible patients for sepsis trials without influencing baseline mortality rate. PMID- 16215368 TI - Clinical and economic consequences of ventilator-associated pneumonia: a systematic review. AB - BACKGROUND: Ventilator-associated pneumonia (VAP) is the most common nosocomial infection in critically ill patients. The clinical and economic consequences of VAP are unclear, with a broad range of values reported in the literature OBJECTIVE: To perform a systematic review to determine the incidence of VAP and its attributable mortality rate, length of stay, and costs. DATA SOURCE: Computerized PUBMED and MEDLINE search supplemented by manual searches for relevant articles, limited to articles published after 1990. STUDY SELECTION: English-language observational studies and randomized trials that provided data on the incidence of VAP were included. Matched cohort studies were included for calculation of attributable mortality rate and length of stay. DATA EXTRACTION: Data were extracted on patient population, diagnostic criteria for VAP, incidence, outcome, type of intensive care unit, and study design. DATA SYNTHESIS: The cumulative incidence of VAP was calculated by combining the results of several studies using standard formulas for combining proportions, in which the weighted average and variance are calculated. Results from studies comparing intensive care unit and hospital mortality due to VAP, additional length of stay, and additional days of mechanical ventilation were pooled using a random effects model, with assessment of heterogeneity. RESULTS: Our findings indicate a) between 10% and 20% of patients receiving >48 hrs of mechanical ventilation will develop VAP; b) critically ill patients who develop VAP appear to be twice as likely to die compared with similar patients without VAP (pooled odds ratio, 2.03; 95% confidence interval, 1.16-3.56); c) patients with VAP have significantly longer intensive care unit lengths of stay (mean = 6.10 days; 95% confidence interval, 5.32-6.87 days); and d) patients who develop VAP incur > or = USD $10,019 in additional hospital costs. CONCLUSIONS: Ventilator-associated pneumonia occurs in a considerable proportion of patients undergoing mechanical ventilation and is associated with substantial morbidity, a two-fold mortality rate, and excess cost. Given these findings, strategies that effectively prevent VAP are urgently needed. PMID- 16215369 TI - Early changes in organ function predict eventual survival in severe sepsis. AB - OBJECTIVE: Early identification and treatment of severe sepsis can significantly reduce mortality rate. We hypothesized that a risk prediction model based on early (baseline to day 1 of study) response to standard care should be significantly related to 28-day survival. DESIGN: Analysis of organ dysfunction data from two placebo-controlled severe sepsis trials (PROWESS and secretory phospholipase A2 inhibitor trials). SETTING: Research laboratory. PATIENTS: The placebo arms of two randomized, double-blind sepsis trials were combined (n = 1036). These patients met criteria for severe sepsis and received supportive standard intensive care and fluid resuscitation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Sequential Organ Failure Assessment (SOFA) scores were calculated daily using the most aberrant physiologic or laboratory variables. Baseline and post-baseline SOFA scores categorized as improved, unchanged, or worsened were used in regression analyses correlating organ dysfunction changes with 28-day mortality. Improvement in cardiovascular (p = .0010), renal (p < .0001), or respiratory (p = .0469) function from baseline to day 1 was significantly related to survival. Odds ratios (95% confidence intervals) associated with improved vs. worsened respiratory, cardiovascular, or renal function before start of day 1 were 0.56 (0.35-0.91), 0.33 (0.18-0.59), and 0.30 (0.17-0.52), respectively. Continued improvement in cardiovascular function before start of day 2 and start of day 3 was associated with further improvement in survival (p <. 0001), with odds ratios of 0.15 (0.06-0.39) and 0.11 (0.04 0.31) for patients who improved compared with those who worsened. No other organ system was retained in the model, and improvement beyond day 1 in any other organ function did not add to the model's predictive power. CONCLUSIONS: These analyses suggest that outcomes for patients with severe sepsis are closely related to early (baseline to day 1 here) improvement, or lack thereof, in organ function. Also, clinical improvement on subsequent days may have little additional impact on the likelihood of survival. PMID- 16215371 TI - Transcutaneous PCO2 monitoring in critically ill adults: clinical evaluation of a new sensor. AB - OBJECTIVE: In critically ill patients, arterial blood gas analysis is the gold standard for evaluating systemic oxygenation and carbon dioxide partial pressure. A new miniaturized carbon dioxide tension Pco2-Spo2 single sensor (TOSCA, Linde Medical Sensors AG, Basel, Switzerland) continuously and noninvasively (transcutaneously) monitors both Paco2 and oxygen saturation by pulse oximetry (Spo2). The present study was designed to investigate the usability and the accuracy of this device in critically ill patients. DESIGN: Prospective clinical investigation. SETTING: A 20-bed, university-affiliated, surgical intensive care unit. PATIENTS: Patients admitted after major surgery, multiple trauma, or septic shock equipped with an arterial catheter. INTERVENTIONS: The heated (42 degrees C) sensor was fixed at the earlobe using an attachment clip. Transcutaneous Pco2 (TcPco2) measurements were correlated with Paco2 values (measured using a blood gas analyzer). In addition, the differences between Paco2 and TcPco2 values were evaluated using the method of Bland-Altman. MEASUREMENTS AND MAIN RESULTS: We studied 55 patients, aged 18-80 (mean 57 +/- 15) yrs. A total of 417 paired measurements were compared. Correlation between TcPco2 and Paco2 was r = .86 (p < .01) in the Paco2 range of 24-101 mm Hg. Mean bias (+/-sd) between the two methods of measurement (Bland-Altman analysis) was 1.2 +/- 6.0 mm Hg with TcPco2 slightly overestimating arterial carbon dioxide tension. Nineteen percent of the measured values were outside of the acceptable clinical range of agreement of +/ 7.5 mm Hg. CONCLUSIONS: The present study suggests that Paco2 can be acceptably assessed by measuring TcPco2 using the TOSCA Pco2-Spo2 sensor. PMID- 16215372 TI - Effect of intracranial pressure monitoring and targeted intensive care on functional outcome after severe head injury. AB - OBJECTIVE: : Intracranial hypertension after severe head injury is associated with case fatality, but there is no sound evidence that monitoring of intracranial pressure (ICP) and targeted management of cerebral perfusion pressure (CPP) improve outcome, despite widespread recommendation by experts in the field. The purpose was to determine the effect of ICP/CPP-targeted intensive care on functional outcome and therapy intensity levels after severe head injury. DESIGN: : Retrospective cohort study with prospective assessment of outcome. SETTING: : Two level I trauma centers in The Netherlands from 1996 to 2001. PATIENTS: : Three hundred thirty-three patients who had survived and remained comatose for >24 hrs, from a total of 685 consecutive severely head-injured adults. INTERVENTIONS: : In center A (supportive intensive care), mean arterial pressure was maintained at approximately 90 mm Hg, and therapeutic interventions were based on clinical observations and computed tomography findings. In center B (ICP/CPP-targeted intensive care), management was aimed at maintaining ICP <20 mm Hg and CPP >70 mm Hg. Allocation to either trauma center was solely based on the site of the accident. MEASUREMENTS AND MAIN RESULTS: : We measured extended Glasgow Outcome Scale after >/=12 months. Patient characteristics were well balanced between the centers. ICP monitoring was used in zero of 122 (0%) and 142 of 211 (67%) patients in centers A and B, respectively. In-hospital mortality rate was 41 (34%) vs. 69 (33%; p = .87). The odds ratio for a more favorable functional outcome following ICP/CPP-targeted therapy was 0.95 (95% confidence interval, 0.62-1.44). This result remained after adjustment for potential confounders. Sedatives, vasopressors, mannitol, and barbiturates were much more frequently used in center B (all p < .01). The median number of days on ventilator support in survivors was 5 (25th-75th percentile, 2-9) in center A vs. 12 (7-19) in center B (p < .001). CONCLUSIONS: : ICP/CPP-targeted intensive care results in prolonged mechanical ventilation and increased levels of therapy intensity, without evidence for improved outcome in patients who survive beyond 24 hrs following severe head injury. PMID- 16215373 TI - Prognostic value of a simple evolving disseminated intravascular coagulation score in patients with severe sepsis. AB - OBJECTIVE: We postulated that the coagulopathy initiated by the inflammatory response to severe sepsis would be reflected by changes in the platelet count and prothrombin time that convey prognostic information. To examine this hypothesis, we looked at the utility of a simple evolving disseminated intravascular coagulation (DIC) score that awarded 1 point for each of the following: a) an absolute platelet count <100 x 10/L; b) a prothrombin time >15.0 secs; c) a 20% decrease in platelets; and d) a >0.3-sec increase in prothrombin time in predicting outcome in patients with severe sepsis. DESIGN: Prospective observational study. SETTING: Intensive care units of university medical center. PATIENTS: Patients were 163 critically ill severe sepsis patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were clinically classified as having capillary leak syndrome (n = 24), multiple organ failure with death from sepsis (n = 37), or multiple organ failure with recovery (n = 57) or as well (n = 45) if they showed rapid improvement in their modified Multiple Organ Dysfunction Syndrome (MODS) score (which did not score for thrombocytopenia). Patients with capillary leak syndrome had the highest Acute Physiology and Chronic Health Evaluation II score, modified MODS, and prothrombin time and the lowest platelet counts, whereas well patients had the most normal values. The simple evolving DIC score increased with worsening clinical class and was associated with worsening organ failure (increased modified MODS). Mortality rate increased from 10% for a simple evolving score of 0 to 73% for a score of 4 (p < .01). Overall, 86% of those with a score < or =1 survived, whereas 85% of those with a score of > or =2 developed multiple organ failure and half of them died from sepsis. CONCLUSIONS: The simple evolving DIC score calculated in the first 48 hrs from two readily available global coagulation markers appears to reflect the severity of the underlying disorder. It can be easily calculated at the bedside and provides useful prognostic information for the patient with severe sepsis. PMID- 16215374 TI - Pharmacodynamics of antimicrobials for the empirical treatment of nosocomial pneumonia: a report from the OPTAMA Program. AB - OBJECTIVE: To compare the probability of achieving specific pharmacodynamic exposures of commonly used intravenous antibiotics for the empirical treatment of nosocomial pneumonia against those pathogens most commonly implicated in the disease. DESIGN: Ten thousand-subject Monte Carlo simulation. SETTING: Research center. SUBJECT: None. INTERVENTIONS: Pharmacodynamic analysis was conducted for the following antimicrobials at standard doses: meropenem, imipenem-cilastatin, ceftazidime, cefepime, piperacillin/tazobactam, and ciprofloxacin. Prevalence of causative pathogens was based on the 2000 SENTRY Antimicrobial Surveillance Study, and minimum inhibitory concentration (MIC) values were obtained using the 2003 US MYSTIC database. The probabilities of each drug and dosing regimen in achieving pharmacodynamic targets were calculated. Bactericidal targets were defined as 40% T>MIC for the carbapenems, 50% T>MIC for other beta-lactams, and an area under the curve (AUC)/MIC ratio of 125 for ciprofloxacin. A sensitivity analysis was performed using two alternate models to determine the impact of varying pathogen prevalence on target attainment. MEASUREMENTS AND MAIN RESULTS: Meropenem and imipenem provided high probabilities of achieving their bactericidal target of 40% T>MIC, with target attainments of 98% for all regimens. At the bactericidal end point of 50% T>MIC, cefepime 2 g every 8 hrs displayed the highest target attainment at 99.9%, followed by cefepime 2 g every 12 hrs, ceftazidime 2 g every 8 hrs, piperacillin/tazobactam 4.5 g every 6 hrs and 3.375 g every 6 hrs, cefepime 1 g every 12 hrs, and ceftazidime 1 g every 8 hrs with target attainments of 95.0%, 92.5%, 92.3%, 91.3%, 90.3%, and 67.9%, respectively. Ciprofloxacin presented the lowest probability of achieving its bactericidal target of an AUC/MIC ratio of 125, with target attainments of 54.7% and 12.0% when given as 400 mg every 8 hrs and 400 mg every 12 hrs, respectively. CONCLUSIONS: Meropenem, imipenem, cefepime, ceftazidime (2 g every 8 hrs), and piperacillin/tazobactam have high probabilities of achieving adequate pharmacodynamic exposures when given for the empirical treatment of nosocomial pneumonia in the absence of methicillin-resistant S. aureus. Ceftazidime 1g every 8 hrs and ciprofloxacin produce low target attainment rates and will not likely result in high clinical success rates when given as monotherapy. PMID- 16215375 TI - Acute respiratory distress syndrome: underrecognition by clinicians and diagnostic accuracy of three clinical definitions. AB - OBJECTIVE: To determine and compare the diagnostic accuracy of three clinical definitions of acute respiratory distress syndrome (ARDS): (1) the American European consensus conference definition; (2) the lung injury score; and (3) a recently developed Delphi definition. A second objective was to determine the accuracy of clinical diagnoses of ARDS made in daily practice. DESIGN: Independent comparison of autopsy findings with the daily status of clinical definitions, constructed with data abstracted retrospectively from medical records. SETTING: Tertiary intensive care unit. PATIENTS: One hundred thirty eight patients from the period 1995 through 2001 who were autopsied after being mechanically ventilated. INTERVENTIONS: Clinical ARDS diagnoses were determined daily without knowledge of autopsy results. Charts were reviewed for any mention of ARDS in the clinical notes. Autopsies were reviewed independently by two pathologists for the presence of diffuse alveolar damage. The sensitivity and specificity of the definitions were determined with use of diffuse alveolar damage at autopsy as the reference standard. MEASUREMENTS AND MAIN RESULTS: Diffuse alveolar damage at autopsy was documented in 42 of 138 cases (30.4%). Only 20 of these 42 patients (47.6%) had any mention of ARDS in their chart. Sensitivities and specificities (95% confidence intervals) were as follows: American-European definition, 0.83 (0.72-0.95), 0.51 (0.41-0.61); lung injury score, 0.74 (0.61-0.87), 0.77 (0.69-0.86); and Delphi definition, 0.69 (0.55 0.83), 0.82 (0.75-0.90). Specificity was significantly higher for both the lung injury score and Delphi definition than for the American-European definition (p < .001 for both), whereas comparisons of sensitivity, which was higher for the American-European definition, were not significantly different (p = .34 and p = .07, respectively). CONCLUSIONS: Acute respiratory distress syndrome appears underrecognized by clinicians in patients who die with this syndrome. In this population, the specificities of existing clinical definitions vary considerably, which may be problematic for clinical trials. PMID- 16215376 TI - Lactate and glucose metabolism in severe sepsis and cardiogenic shock. AB - OBJECTIVE: To evaluate the relative importance of increased lactate production as opposed to decreased utilization in hyperlactatemic patients, as well as their relation to glucose metabolism. DESIGN: Prospective observational study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: Seven patients with severe sepsis or septic shock, seven patients with cardiogenic shock, and seven healthy volunteers. INTERVENTIONS: C-labeled sodium lactate was infused at 10 micromol/kg/min and then at 20 micromol/kg/min over 120 mins each. H-labeled glucose was infused throughout. MEASUREMENTS AND MAIN RESULTS: Baseline arterial lactate was higher in septic (3.2 +/- 2.6) and cardiogenic shock patients (2.8 +/ 0.4) than in healthy volunteers (0.9 +/- 0.20 mmol/L, p < .05). Lactate clearance, computed using pharmacokinetic calculations, was similar in septic, cardiogenic shock, and controls, respectively: 10.8 +/- 5.4, 9.6 +/- 2.1, and 12.0 +/- 2.6 mL/kg/min. Endogenous lactate production was determined as the initial lactate concentration multiplied by lactate clearance. It was markedly enhanced in the patients (septic 26.2 +/- 10.5; cardiogenic shock 26.6 +/- 5.1) compared with controls (11.2 +/- 2.7 micromol/kg/min, p < .01). C-lactate oxidation (septic 54 +/- 25; cardiogenic shock 43 +/- 16; controls 65 +/- 15% of a lactate load of 10 micromol/kg/min) and transformation of C-lactate into C glucose were not different (respectively, 15 +/- 15, 9 +/- 18, and 10 +/- 7%). Endogenous glucose production was markedly increased in the patients (septic 14.8 +/- 1.8; cardiogenic shock 15.0 +/- 1.5) compared with controls (7.2 +/- 1.1 micromol/kg/min, p < .01) and was not influenced by lactate infusion. CONCLUSIONS: In patients suffering from septic or cardiogenic shock, hyperlactatemia was mainly related to increased production, whereas lactate clearance was similar to healthy subjects. Increased lactate production was concomitant to hyperglycemia and increased glucose turnover, suggesting that the latter substantially influences lactate metabolism during critical illness. PMID- 16215377 TI - Recombinant activated factor VII for refractory bleeding after cardiac surgery--a retrospective analysis of safety and efficacy. AB - OBJECTIVE: Analysis of safety and efficacy of recombinant activated factor VII (rFVIIa) used as the last resort for refractory bleeding after cardiac surgery. DESIGN: Retrospective cohort analysis and matched pairs analysis with historic controls were performed. In the rFVIIa group, which also received conventional hemostatic therapy, data were collected for a median of 14 hrs from admission to the intensive care unit (ICU) to the administration of rFVIIa and for the following 24 hrs. In the control group, which received only conventional hemostatic therapy, data were collected for 14 and then for 24 hrs after admission to the ICU. SETTING: University hospital. PATIENTS: Twenty-four patients matched with historic controls. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: No thromboembolic complications were observed in the rFVIIa group. Blood loss and transfusion requirements were significantly reduced in the period after the administration of rFVIIa. However, in the 24-hr period after rFVIIa administration, blood loss (p = .140) and transfusion of packed red blood cells (p = .442) and fresh frozen plasma (p = .063) were not different between the rFVIIa and control groups. Platelet concentrates (p = .004) were transfused less in the control group. Mortality and 6-month survival rates were not different between the groups. CONCLUSIONS: When used as a last resort, rFVIIa was safe but not incrementally efficacious over conventional hemostatic therapy. PMID- 16215378 TI - Red blood cell 2,3-diphosphoglycerate concentration and in vivo P50 during early critical illness. AB - OBJECTIVE: To measure red blood cell 2,3-diphosphoglycerate (RBC 2,3-DPG) concentrations in early critical illness; to investigate factors associated with high or low RBC 2,3-DPG levels; to calculate in vivo P50 in patients with early critical illness; and to explore the relationship between RBC 2,3-DPG and intensive care mortality. DESIGN: Prospective cohort study. SETTING: General medical-surgical intensive care unit (ICU) of a major Scottish teaching hospital. PATIENTS: One-hundred eleven critically ill patients during the first 24 hrs in the ICU with no history of chronic hematologic disorders or RBC transfusion within 24 hrs and 34 age- and sex-matched healthy reference subjects. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We measured RBC 2,3-DPG concentration, plasma biochemistry values, and arterial blood gas parameters. On average, RBC 2,3-DPG was lower among critically ill patients than controls (mean [sd], 14.1 [6.3] vs. 16.7 [3.7] mumol/g hemoglobin; p = .004) and had a wider range of values (patients, 3.2-32.5 mumol/g hemoglobin; reference group, 9.1 24.3). Regression analysis indicated a strong independent association between plasma pH and RBC 2,3-DPG (B, 32.15 [95% confidence interval, 19.07-46.22], p < .001) and a weak association with plasma chloride (B, -0.196 [95% confidence interval, -0.39 to -0.01], p = .044) but not with hemoglobin or other measured biochemical parameters. The mean calculated in vivo P50 level was normal (3.8 kPa) but varied widely among patients (range, 2.0-5.5 kPa). RBC 2,3-DPG concentration was similar for ICU survivors and nonsurvivors. CONCLUSIONS: RBC 2,3-DPG concentrations vary widely among critically ill patients. Acidosis is associated with lower RBC 2,3-DPG concentrations, but anemia is not associated with a compensatory increase in RBC 2,3-DPG early in critical illness. Lower RBC 2,3-DPG concentrations during the first 24 hrs of intensive care are not associated with higher ICU mortality. PMID- 16215379 TI - Open lung ventilation improves functional residual capacity after extubation in cardiac surgery. AB - OBJECTIVE: After cardiac surgery, functional residual capacity (FRC) after extubation is reduced significantly. We hypothesized that ventilation according to the open lung concept (OLC) attenuates FRC reduction after extubation. DESIGN: A prospective, single-center, randomized, controlled clinical study. SETTING: Cardiothoracic operating room and intensive care unit of a university hospital. PATIENTS: Sixty-nine patients scheduled for elective coronary artery bypass graft and/or valve surgery with cardiopulmonary bypass. INTERVENTIONS: Before surgery, patients were randomly assigned to three groups: (1) conventional ventilation (CV); (2) OLC, started after arrival in the intensive care unit (late open lung); and (3) OLC, started directly after intubation (early open lung). In both OLC groups, recruitment maneuvers were applied until Pao2/Fio2 was >375 Torr (50 kPa). No recruitment maneuvers were applied in the CV group. MEASUREMENTS AND MAIN RESULTS: FRC was measured preoperatively and 1, 3, and 5 days after extubation. Peripheral hemoglobin saturation (Spo2) was measured daily till the third day after extubation while the patient was breathing room air. Hypoxemia was defined by an Spo2 value < or =90%. Averaged over the 5 postoperative days, FRC was significantly higher in the early open lung group and tended to be higher in the late open lung group, in comparison with the CV group (mean +/- sem: CV, 1.8 +/- 0.1; late open lung,1.9 +/- 0.1; and early open lung, 2.2 +/- 0.1l). In the CV group, 37% of the patients were hypoxic on the third day after extubation, compared with none of the patients in both OLC groups. CONCLUSIONS: After cardiac surgery, earlier application of OLC resulted in a significantly higher FRC and fewer episodes of hypoxemia than with CV after extubation. PMID- 16215380 TI - Impact of whole-body rehabilitation in patients receiving chronic mechanical ventilation. AB - OBJECTIVE: To evaluate the prevalence and magnitude of weakness in patients receiving chronic mechanical ventilation and the impact of providing aggressive whole-body rehabilitation on conventional weaning variables, muscle strength, and overall functional status. DESIGN: Retrospective analysis of 49 consecutive patients. SETTING: Multidisciplinary ventilatory rehabilitation unit in an academic medical center. PATIENTS: Forty-nine consecutive chronic ventilator dependent patients referred to a tertiary care hospital ventilator rehabilitation unit. INTERVENTIONS: None. MEASUREMENT AND MAIN RESULTS: Patients were 58 +/- 7 yrs old with multiple etiologies for respiratory failure. On admission, all patients were bedridden and had severe weakness of upper and lower extremities measured by a 5-point muscle strength score and a 7-point Functional Independence Measurement. Postrehabilitation, patients had increases in upper and lower extremity strength (p < .05) and were able to stand and ambulate. All weaned from mechanical ventilation, but three required subsequent intermittent support. Six patients died before hospital discharge. Upper extremity strength on admission inversely correlated with time to wean from mechanical ventilation (R = .72, p < .001). CONCLUSIONS: : Patients receiving chronic ventilation are weak and deconditioned but respond to aggressive whole-body and respiratory muscle training with an improvement in strength, weaning outcome, and functional status. Whole-body rehabilitation should be considered a significant component of their therapy. PMID- 16215381 TI - Drotrecogin alfa (activated) treatment in severe sepsis from the global open label trial ENHANCE: further evidence for survival and safety and implications for early treatment. AB - OBJECTIVE: To provide further evidence for the efficacy and safety of drotrecogin alfa (activated) treatment in severe sepsis. DESIGN: Single-arm, open-label, trial of drotrecogin alfa (activated) treatment in severe sepsis patients. Enrollment began in March 2001 and day-28 follow-up completed in January 2003. SETTING: ENHANCE took place in 25 countries at 361 sites. PATIENTS: Patients with known or suspected infection, three or four systemic inflammatory response syndrome criteria, and one or more sepsis-induced organ dysfunctions. Of 2,434 adults entered, 2,378 received drotrecogin alfa (activated), and of these, 2,375 completed the protocol. INTERVENTIONS: Drotrecogin alfa (activated) was infused at a dose of 24 mug/kg/hr for 96 hrs. MEASUREMENTS AND MAIN RESULTS: The 28-day all-cause mortality approximated that observed in PROWESS (25.3% vs. 24.7%). Although patients in ENHANCE had increased serious bleeding rates compared with patients in the drotrecogin alfa (activated) arm of PROWESS (during infusion, 3.6% vs. 2.4%; postinfusion, 3.2% vs. 1.2%; 28-day, 6.5% vs. 3.5%), increased postinfusion bleeding suggested a higher background bleeding rate. Intracranial hemorrhage was more common in ENHANCE than PROWESS (during infusion, 0.6% vs. 0.2%; 28-day, 1.5% vs. 0.2%). The incidence of fatal intracranial hemorrhage was the same during infusion (0.2%) and higher at 28 days (0.5% vs. 0.2%). ENHANCE patients treated within 0-24 hrs from their first sepsis-induced organ dysfunction had lower observed mortality rate than those treated after 24 hrs (22.9% vs. 27.4%, p = .01). CONCLUSIONS: ENHANCE provides supportive evidence for the favorable benefit/risk ratio observed in PROWESS and suggests that more effective use of drotrecogin alfa (activated) might be obtained by initiating therapy earlier. PMID- 16215382 TI - Gut luminal microdialysis of glycerol as a marker of intestinal ischemic injury and recovery. AB - OBJECTIVE: To evaluate microdialysis as a method to assess different degrees of intestinal damage and recovery during ischemia and reperfusion; to evaluate information obtained from microdialysis catheters in the peritoneum, the gut wall, and the gut lumen. DESIGN: Randomized, controlled animal experiment. SETTING: University laboratory animal center. SUBJECTS: Twenty-seven domestic pigs. INTERVENTIONS: The superior mesenteric artery was cross-clamped for 60 mins (n = 14) or 120 mins (n = 10) followed by 2 or 4 hrs of reperfusion. Three pigs served as controls. MEASUREMENTS AND MAIN RESULTS: Intestinal mucosal integrity was assessed by morphometry, adenosine triphosphate in the gut wall, and permeability of C-polyethylene glycol. Lactate, glycerol, pyruvate, and glucose were measured by microdialysis. Changes in adenosine triphosphate, permeability, or lactate did not correlate to different extents of intestinal damage caused by 60 or 120 mins of ischemia. During the reperfusion period, pigs with 60 mins of intestinal ischemia showed a faster recovery of these variables than pigs with 120 mins of intestinal ischemia. Glycerol increased with increasing duration of the ischemic insult. After 60 mins of intestinal ischemia, glycerol in the gut lumen decreased toward baseline but remained high after 120 mins of intestinal ischemia. There was a good correlation between gut luminal glycerol and recovery of mucosal damage throughout the reperfusion period. In the peritoneal cavity, both glycerol and lactate decreased to baseline relatively shortly after onset of reperfusion independent of the duration of intestinal ischemia. CONCLUSIONS: Microdialysis of glycerol provides information about the extent and severity of intestinal damage after ischemia and about the ensuing recovery. The gut lumen is to be preferred as a site for placement of microdialysis catheters. PMID- 16215383 TI - Bilateral lavage with diluted surfactant improves lung function after unilateral lung contusion in pigs. AB - OBJECTIVE: This study evaluates the effects of bronchoalveolar lavage with diluted surfactant on unilateral lung contusion-induced lung dysfunction. DESIGN: Randomized prospective animal study. SETTING: An animal laboratory. SUBJECTS: Twenty adult pigs, weighing 25-35 kg. INTERVENTIONS: Animals were randomly assigned to controls and surfactant treatment. Bilateral lavage with surfactant treatment began 30 mins after unilateral lung contusion. Then 25 mg/kg of body weight diluted Curosurf (5 mg/mL) was applied in a volume of 5 mL/kg of body weight. Observation time was 8 hrs postinjury. MEASUREMENTS AND MAIN RESULTS: The Pao2/Fio2 ratio fell from 500 to 250 and then recovered gradually in controls and surfactant-treated pigs. After another 4 hrs, the Pao2/Fio2 ratio deteriorated again in controls, but not in surfactant-treated animals. Total compliance fell by 50% after injury but was completely restored by surfactant treatment. Lung contusion increased the median number of neutrophils in bronchoalveolar lavage fluid from 2% to 30% of total cells and peaked >60% at 480 mins in the contused lungs of control pigs. Surfactant-treated pigs had 40% neutrophils at 480 mins without reaching significant difference to controls. The leukocyte neutral proteinase inhibitor increased to 500 ng/mL at 30 mins postinjury in the contused lungs and increased to 2000 ng/mL after surfactant treatment. CONCLUSIONS: Bilateral bronchoalveolar lavage with diluted surfactant can effectively improve lung function after experimental unilateral lung contusion in pigs. PMID- 16215384 TI - Infusion of the beta-adrenergic blocker esmolol attenuates myocardial dysfunction in septic rats. AB - OBJECTIVE: Since beta-blocker therapy is known to be effective in patients with an injured heart, such as infarction, we designed the present study to examine the protective effects of infusion of the beta1-selective blocker esmolol on myocardial function in peritonitis-induced septic rats using an isolated working heart preparation. DESIGN: Randomized animal study. SETTING: University research laboratory. SUBJECTS: Thirty-one rats treated with cecal ligation and perforation to evoke peritonitis. INTERVENTIONS: After cecal ligation and perforation, rats were randomly allocated to the control group (normal saline 2 mL/hr, n = 11), low dose esmolol group (10 mg/kg/hr, n = 10), or high-dose esmolol group (20 mg/kg/hr, n = 10). After obtaining blood samples for measurement of arterial lactate and tumor necrosis factor-alpha at 24 hrs, we assessed cardiac output, myocardial oxygen consumption, and cardiac efficiency (cardiac output x peak systolic pressure/myocardial oxygen consumption) at various preloads in an isolated perfused heart preparation. MEASUREMENTS AND MAIN RESULTS: Esmolol infusion did not cause an elevation of arterial lactate levels but reduced tumor necrosis factor-alpha concentrations vs. the control group (p < .05). Both cardiac output and cardiac efficiency in the esmolol-treated rats were significantly higher throughout the study periods vs. the control group (p < .05). CONCLUSIONS: Esmolol infusion in sepsis improved oxygen utilization of myocardium and preserved myocardial function. PMID- 16215385 TI - Cardioprotective effects of acute isovolemic hemodilution in a rat model of transient coronary occlusion. AB - OBJECTIVES: Following isovolemic hemodilution (AIH), lowering blood viscosity induces acceleration of erythrocyte velocity resulting in improved tissue oxygen delivery. Using a rat model of myocardial infarct, we tested the hypothesis that AIH would attenuate myocardial damage due to transient coronary occlusion. DESIGN: Prospective, randomized, and controlled animal study. SETTING: Animal research laboratory in a university hospital. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: All rats were subjected to 30 mins of left coronary artery occlusion followed by 48 hrs of reperfusion. Before the ischemic period, the anesthetized rats were randomly allocated to undergo either 15 mins of waiting (controls) or AIH to achieve a hematocrit of 30% (AIH-CO) by stepwise blood withdrawal and isovolemic compensation with 6% hydroxyethylstarch 200-0.5. MEASUREMENTS AND MAIN RESULTS: Hemodynamic variables were comparable in the two groups, except for higher indexes of stroke volume in the AIH-CO group. During coronary occlusion and the reperfusion period, AIH resulted in a lower incidence of fatal ventricular tachyarrhythmia (17% vs. 50% in control group, p < .05) and higher survival at 48 hrs of postreperfusion (83% vs. 42%, p < .05).Preischemic hemodilution significantly attenuated myocardial damage as shown by lower release of cardiac troponin I and reduction in myocardial infarct size as measured by tetrazolin staining. Histologic examination revealed no difference regarding peri ischemic infiltration with neutrophil granulocytes. CONCLUSIONS: Our data provide the first experimental demonstration that preischemic moderate AIH confers cardioprotection and improves survival in a rat model of myocardial infarct. PMID- 16215386 TI - Rosiglitazone, a peroxisome proliferator-activated receptor-gamma agonist, reduces acute lung injury in endotoxemic rats. AB - OBJECTIVE: Rosiglitazone, a potent agonist of peroxisome proliferator-activated receptor (PPAR)-gamma, exerts anti-inflammatory effects in vitro and in vivo. This study was designated to determine the effects of rosiglitazone on endotoxin induced acute lung injury in rats. DESIGN: Prospective, experimental study. SETTING: University research laboratory. SUBJECTS: Thirty-six male Wistar rats. INTERVENTIONS: All the animals were randomly assigned to one of six groups (n = 6 per group) and were given either lipopolysaccharide (6 mg/kg intravenously) or saline, pretreated with rosiglitazone (0.3 mg/kg intravenously) or vehicle (10% dimethyl sulphoxide) 30 mins before lipopolysaccharide. The selective PPAR-gamma antagonist GW9662 (0.3 mg/kg intravenously) or its vehicle (10% dimethyl sulphoxide) was given 20 mins before rosiglitazone. MEASUREMENTS AND MAIN RESULTS: Endotoxemia for 4 hrs induced evident lung histologic injury and edema, both of which were significantly attenuated by rosiglitazone pretreatment. The protective effects of rosiglitazone were correlated with the reduction by 71% of the increase of myeloperoxidase activity and the reduction by 84% of the increase of malondialdehyde in the lung tissue. The pulmonary hyperproduction of nitric oxide was reduced by 82% of the increase related to lipopolysaccharide challenge. Pretreatment with rosiglitazone also markedly suppressed lipopolysaccharide induced expression of inducible nitric oxide synthase messenger RNA and protein in the lung, as demonstrated by reverse transcription-polymerase chain reaction or Western blot analysis. Immunohistochemical analysis revealed that rosiglitazone inhibited the formation of nitrotyrosine, a marker for peroxynitrite reactivity, in the lung tissue. In addition, the specific PPAR gamma antagonist GW9662 antagonized the effects of rosiglitazone. CONCLUSIONS: This study provides evidence, for the first time, that the PPAR-gamma agonist rosiglitazone significantly reduces endotoxin-induced acute lung injury in rats. PMID- 16215387 TI - Interleukin-10 aerosol reduces proinflammatory mediators in bronchoalveolar fluid of endotoxemic rat. AB - OBJECTIVE: We aimed to clarify if inhaled interleukin (IL)-10 attenuates pulmonary and systemic inflammation as indicated by reduced content of proinflammatory mediators in bronchoalveolar lavage fluid (BALF) and plasma in experimental endotoxemia in the rat. DESIGN: Laboratory experiment. SETTING: University research institute. SUBJECTS: Anesthetized, ventilated rats (sd, 550 +/- 50 g). INTERVENTIONS: Rats were randomly treated as follows: Nebulized IL-10 (calculated deposition fraction, 0.1 microg/lung) was administered in eight rats before infusion of lipopolysaccharide (5 mg/kg, intravenously). Eight animals received the same insult with no further treatment. Eight rats served as controls without endotoxemia but with aerosolized saline. MEASUREMENTS AND MAIN RESULTS: BALF and plasma levels of tumor necrosis factor (TNF)-alpha, IL-1beta, IL-6, interferon (IFN)-gamma, and RANTES were analyzed. Alveolar macrophages were cultured ex vivo for nitrite assay. In those animals treated with IL-10-aerosol, BALF levels of proinflammatory cytokines were reduced significantly compared with animals without IL-10 therapy (TNF-alpha, -87%; IL-1beta, -73%; IL-6, -44%; IFN gamma, -39%; RANTES, -84%). In addition, nitrite release from cultured alveolar macrophages was suppressed by IL-10 inhalation (-96%). With the exception of TNF alpha, similar results were observed for plasma levels of proinflammatory cytokines. CONCLUSIONS: The present data indicate that nebulized IL-10 reached the lungs in therapeutic effective concentrations and elicited anti-inflammatory effects on immunocompetent cells that are comparable to those already known from its intravenous administration in experimental endotoxemia. PMID- 16215388 TI - Carbon monoxide, but not endothelin-1, plays a major role for the hepatic microcirculation in a murine model of early systemic inflammation. AB - OBJECTIVE: Endothelin-1 and carbon monoxide play a major role in the regulation of liver microcirculation in numerous disease states. During sepsis and endotoxemia, elevated formation of endothelin-1 results in reduced sinusoidal blood flow. However, the role of carbon monoxide and endothelin-1 and its receptors endothelin receptor A and endothelin receptor B in the deranged liver microcirculation during early systemic inflammation remains unclear. DESIGN: Prospective, randomized, controlled experiment. SETTING: University animal laboratory. SUBJECTS: Male C57/BL6 mice, weighing 23-27 g. INTERVENTIONS: To induce a systemic inflammation, mice were treated with 1 hr of bilateral hind limb ischemia followed by 3 hrs or 6 hrs of reperfusion. Animals were randomly exposed to the nonselective endothelin receptor antagonist Ro-61-6612 (Tezosentan) and/or a continuous endothelin-1 infusion. Different animals were randomized to methylene chloride gavage or carbon monoxide inhalation during the reperfusion period. MEASUREMENTS AND MAIN RESULTS: After ischemia/reperfusion, endothelin-1 plasma concentrations, endothelin-1 messenger RNA expression, and endothelin receptor A and B messenger RNA expression revealed no significant changes when compared with sham animals. After 6 hrs of ischemia/reperfusion, hepatic microcirculatory variables (sinusoidal density, sinusoidal diameter, and red blood cell velocity) deteriorated. Tezosentan after 6 hrs of ischemia/reperfusion did not improve the liver microcirculation, whereas the continuous infusion of endothelin-1 after 6 hrs of ischemia/reperfusion further impaired sinusoidal blood flow. Tezosentan treatment did not produce any alterations in hepatocellular injury or hepatic redox status when compared with the untreated animals receiving 6 hrs of ischemia/reperfusion. Animals receiving 6 hrs of ischemia/reperfusion and exposed to methylene chloride gavage or inhaled carbon monoxide during limb reperfusion showed significantly improved microcirculatory variables, hepatic redox status, and attenuated hepatocellular injury. CONCLUSIONS: These data suggest that endothelin-1 and the endothelin receptors A and B are not responsible for the observed hepatic microcirculatory and cellular dysfunction during early systemic inflammation, but exposure to exogenous carbon monoxide protected the hepatic microcirculation and improved the impaired hepatic cellular integrity and the hepatocellular redox status. PMID- 16215389 TI - Effect of increased cardiac output on hepatic and intestinal microcirculatory blood flow, oxygenation, and metabolism in hyperdynamic murine septic shock. AB - OBJECTIVE: Septic shock-associated organ dysfunction is attributed to derangements of microcirculatory perfusion and/or impaired cellular oxygen utilization. The hepatosplanchnic organs are regarded to play a pivotal role in the pathophysiology of sepsis-related organ failure. In a murine model of septic shock, we tested the hypothesis whether achieving normotensive, hyperdynamic hemodynamics characterized by a sustained increase in cardiac output would allow maintenance of regional microvascular perfusion and oxygenation and, thus, hepatic metabolic capacity. DESIGN: Prospective, controlled, randomized animal study. SETTING: University animal research laboratory. SUBJECTS: Male C57Bl/6 mice. INTERVENTIONS: Fifteen hours after sham operation (n = 11) or cecal ligation and puncture (CLP) (n = 9), mice were anesthetized, mechanically ventilated, and instrumented (central venous and left ventricular pressure conductance catheter, portal vein and superior mesenteric artery ultrasound flow probes). Animals received continuous intravenous hydroxyethylstarch and norepinephrine to achieve normotensive and hyperdynamic hemodynamics, and glucose was infused to maintain normoglycemia. MEASUREMENTS AND MAIN RESULTS: Measurements were recorded 18, 21, and 24 hrs post-CLP. In CLP mice, titration of hemodynamic targets were affiliated superior mesenteric artery and portal vein flow. Using a combined laser-Doppler flowmetry and remission spectrophotometry probe, we found well-maintained gut and liver capillary perfusion as well as intestinal microcirculatory hemoglobin oxygen saturation, whereas hepatic microcirculatory hemoglobin oxygen saturation was even increased. At 24 hrs post CLP, the rate of de novo gluconeogenesis as derived from hepatic C-glucose isotope enrichment after continuous intravenous 1,2,3,4,5,6-C6-glucose infusion (condensation biosynthesis modeling after gas chromatography-mass spectrometry isotope measurements) was similar in the two experimental groups. CONCLUSIONS: During murine septic shock achieving normotensive hyperdynamic hemodynamics with fluid resuscitation and norepinephrine, exogenous glucose requirements together with the lack of norepinephrine-induced increase in the rate of gluconeogenesis mirror impaired metabolic capacity of the liver despite well-maintained hepatosplanchnic microvascular perfusion and oxygenation. PMID- 16215390 TI - Effects of norepinephrine on static and dynamic preload indicators in experimental hemorrhagic shock. AB - OBJECTIVE: To investigate the effect of norepinephrine on static (right atrial pressure, pulmonary artery occlusion pressure ) and dynamic (pulse pressure variation and arterial systolic pressure variation) preload indicators in experimental hemorrhagic shock. DESIGN: Prospective controlled experimental study. SETTING: Animal research laboratory. SUBJECTS: Six anesthetized and mechanically ventilated dogs. INTERVENTIONS: Dogs were instrumented for measurement of arterial blood pressure, pulmonary artery catheter derived variables including right atrial pressure, pulmonary artery occlusion pressure, and cardiac output. Simultaneously, pulse pressure variation and systolic pressure variation were calculated. Pulse pressure variation is the difference between the maximal and the minimal value of pulse pressure divided by the mean of the two values and is expressed as a percentage. Systolic pressure variation is the difference between the maximal and the minimal systolic pressure and is expressed as an absolute value. After baseline measurements, hemorrhagic shock was induced by a stepwise cumulative blood withdrawal of 35 mL.kg of body weight. A second set of hemodynamic measurement was made 30 mins after bleeding. The third set was made 30 mins later under norepinephrine. MEASUREMENTS AND MAIN RESULTS: Mean arterial pressure and cardiac output decreased after hemorrhage (p < .05), whereas right atrial pressure and pulmonary artery occlusion pressure remained unchanged. Baseline pulse pressure variation and systolic pressure variation increased significantly with hemorrhage, from 12% (9%) to 28% (11.5%) (p < .001) and from 12.5 (6.5) to 21 (8.2) mm Hg (p < .05), respectively. Norepinephrine induced a significant increase of cardiac output and a significant decrease of pulse pressure variation and systolic pressure variation but did not significantly change right atrial pressure or pulmonary artery occlusion pressure values. Stroke volume was correlated to pulse pressure variation and systolic pressure variation but was not correlated to right atrial pressure or pulmonary artery occlusion pressure. CONCLUSION: Our study confirms the superiority of dynamic variables (pulse pressure variation and systolic pressure variation) over static ones (right atrial pressure and pulmonary artery occlusion pressure) in assessing cardiac preload changes in hemorrhagic shock. However, norepinephrine could significantly reduce the value of these dynamic variables and mask a true intravascular volume deficit possibly by shifting blood from unstressed to stressed volume. PMID- 16215391 TI - Evaluation of a glycine-rich amino acid solution for parenteral nutrition in endotoxemic rats. AB - OBJECTIVE: It has been shown recently that high amounts of glycine might have some pharmacologic effects (reduction of injury and mortality in endotoxemic rats), but its effects on the nutritional status and protein metabolism during injury are still unknown. The aim of this study was to compare the nutritional effects of a glycine-rich amino acid solution for parenteral nutrition (AFD) with a standard one (Vintene) (glycine, 15 vs. 9 g/L) in endotoxemic rats. DESIGN: Laboratory investigation. SETTING: University laboratory. SUBJECTS: Male Wistar rats (198 +/- 11 g). INTERVENTIONS: Rats were operated to receive total parenteral nutrition (250 kcal/kg/day, 2 g N/kg/day) with amino acids supplied by either AFD (n = 9) or Vintene (V, n = 6). One day after surgery, corresponding to day 0 of the experiment and to the first day of full-strength total parenteral nutrition, the AFD and V group rats received an endotoxemic shock by intraperitoneal injection of lipopolysaccharide (Escherichia coli, 8 mg/kg). The rats were then studied over 3 days and compared with a healthy ad libitum-fed group (AL, n = 10). MEASUREMENTS AND MAIN RESULTS: The rats were weighed and urine was collected daily to determine nitrogen balance and 3-methylhistidine excretion. On day 3, the thymus, spleen, liver, intestinal mucosa, and muscles were weighed, and amino acids from plasma and tissues were analyzed. Lipopolysaccharide caused the classic endotoxemic shock, of similar intensity in the V and AFD groups (V and AFD not equal AL, p < .05): no weight gain, decreased nitrogen balance (day 3, AL 558 +/- 21, V 83 +/- 28, AFD 123 +/- 25 mg N/day), increased urinary 3-methylhistidine/creatinine excretion (day 3, AL 51 +/- 2, V 91 +/- 13, AFD 87 +/- 14 mumol/mmol), soleus (V -15% and AFD -26 % vs. AL) and thymus atrophy (V -36% and AFD -33%), and spleen hypertrophy (V 51% and AFD 83%). Compared with V solution, AFD has a reduced content of some essential amino acids and proline and an elevated content of glycine, aspartate, and glutamate. These differences were not reflected in tissue or plasma amino acids, except for plasma glycine, which in the AFD group was restored to the level of the AL group (AL 426 +/- 12 and AFD 379 +/- 50 vs. V 251 +/- 31 mumol/L, p < .05). CONCLUSIONS: In endotoxemic rats, the nutritional effects of a glycine-rich AFD solution are similar to those of a standard amino acid solution for parenteral nutrition. PMID- 16215393 TI - Acute secondary adrenal insufficiency after traumatic brain injury: a prospective study. AB - OBJECTIVE: To determine the prevalence, time course, clinical characteristics, and effect of adrenal insufficiency (AI) after traumatic brain injury (TBI). DESIGN: Prospective intensive care unit-based cohort study. SETTING: Three level 1 trauma centers. PATIENTS: A total of 80 patients with moderate or severe TBI (Glasgow Coma Scale score, 3-13) and 41 trauma patients without TBI (Injury Severity Score, >15) enrolled between June 2002 and November 2003. MEASUREMENTS: Serum cortisol and adrenocorticotropic hormone levels were drawn twice daily for up to 9 days postinjury; AI was defined as two consecutive cortisols of < or =15 microg/dL (25th percentile for extracranial trauma patients) or one cortisol of < 5 microg/dL. Principal outcome measures included: injury characteristics, hemodynamic data, usage of vasopressors, metabolic suppressive agents (high-dose pentobarbital and propofol), etomidate, and AI status. MAIN RESULTS: AI occurred in 42 TBI patients (53%). Adrenocorticotropic hormone levels were lower at the time of AI (median, 18.9 vs. 36.1 pg/mL; p = .0001). Compared with patients without AI, those with AI were younger (p = .01), had higher injury severity (p = .02), had a higher frequency of early ischemic insults (hypotension, hypoxia, severe anemia) (p = .02), and were more likely to have received etomidate (p = .049). Over the acute postinjury period, patients with AI had lower trough mean arterial pressure (p = .001) and greater vasopressor use (p = .047). Mean arterial pressure was lower in the 8 hrs preceding a low (< or =15 microg/dL) cortisol level (p = .003). There was an inverse relationship between cortisol levels and vasopressor use (p = .0005) and between cortisol levels within 24 hrs of injury and etomidate use (p = .002). Use of high-dose propofol and pentobarbital was strongly associated with lower cortisol levels (p < .0001). CONCLUSIONS: Approximately 50% of patients with moderate or severe TBI have at least transient AI. Younger age, greater injury severity, early ischemic insults, and the use of etomidate and metabolic suppressive agents are associated with AI. Because lower cortisol levels were associated with lower blood pressure and higher vasopressor use, consideration should be given to monitoring cortisol levels in intubated TBI patients, particularly those receiving high-dose pentobarbital or propofol. A randomized trial of stress-dose hydrocortisone in TBI patients with AI is underway. PMID- 16215392 TI - Mechanisms of decreased intestinal epithelial proliferation and increased apoptosis in murine acute lung injury. AB - OBJECTIVES: The aim of this study was to determine the effects of acute lung injury on the gut epithelium and examine mechanisms underlying changes in crypt proliferation and apoptosis. The relationship between severity and timing of lung injury to intestinal pathology was also examined. DESIGN: Randomized, controlled study. SETTING: University research laboratory. SUBJECTS: Genetically inbred mice. INTERVENTIONS: Following induction of acute lung injury, gut epithelial proliferation and apoptosis were assessed in a) C3H/HeN wild-type and C3H/HeJ mice, which lack functional Toll-like receptor 4 (n = 17); b) C57Bl/6 mice that received monoclonal anti-tumor necrosis factor-alpha or control antibody (n = 22); and c) C57Bl/6 wild-type and transgenic mice that overexpress Bcl-2 in their gut epithelium (n = 21). Intestinal epithelial proliferation and death were also examined in animals with differing degrees of lung inflammation (n = 24) as well as in a time course analysis following a fixed injury (n = 18). MEASUREMENTS AND MAIN RESULTS: Acute lung injury caused decreased proliferation and increased apoptosis in crypt epithelial cells in all animals studied. C3H/HeJ mice had higher levels of proliferation than C3H/HeN animals without additional changes in apoptosis. Anti-tumor necrosis factor-alpha antibody had no effect on gut epithelial proliferation or death. Overexpression of Bcl-2 did not change proliferation despite decreasing gut apoptosis. Proliferation and apoptosis were not correlated to severity of lung injury, as gut alterations were lost in mice with more severe acute lung injury. Changes in both gut epithelial proliferation and death were apparent within 12 hrs, but proliferation was decreased 36 hrs following acute lung injury while apoptosis returned to normal. CONCLUSIONS: Acute lung injury causes disparate effects on crypt proliferation and apoptosis, which occur, at least in part, through differing mechanisms involving Toll-like receptor 4 and Bcl-2. Severity of lung injury does not correlate with perturbations in proliferation or death in the gut epithelium, and acute lung injury-induced changes in intestinal epithelial proliferation persist longer than those in apoptosis. PMID- 16215394 TI - Effects of positive end-expiratory pressure on regional cerebral blood flow, intracranial pressure, and brain tissue oxygenation. AB - OBJECTIVE: Acute respiratory dysfunction frequently occurs following severe aneurysmal subarachnoid hemorrhage requiring positive end-expiratory pressure (PEEP) ventilation to maintain adequate oxygenation. High PEEP levels, however, may negatively affect cerebral perfusion. The goal of this study was, to examine the influence of various PEEP levels on intracranial pressure, brain tissue oxygen tension, regional cerebral blood flow, and systemic hemodynamic variables. DESIGN: Animal research and clinical intervention study. SETTING: Surgical intensive care unit of a university hospital. SUBJECTS AND PATIENTS: Experiments were carried out in five healthy pigs, followed by a clinical investigation of ten patients suffering subarachnoid hemorrhage. INTERVENTIONS: Under continuous monitoring of intracranial pressure, brain tissue oxygen tension, regional cerebral blood flow, mean arterial pressure, and cardiac output, PEEP was applied in increments of 5 cm H2O from 5 to 25 cm H2O in the experimental part and from baseline to 20 cm H2O in the clinical part. MEASUREMENTS AND MAIN RESULTS: In animals, high PEEP levels had no adverse effect on intracranial pressure, brain tissue oxygen tension, or regional cerebral blood flow. In patients with severe subarachnoid hemorrhage, stepwise elevation of PEEP resulted in a significant decrease of mean arterial pressure and regional cerebral blood flow. Analyses of covariance revealed that these changes of regional cerebral blood flow depended on mean arterial pressure changes as a result of a disturbed cerebrovascular autoregulation. Consequently, normalization of mean arterial pressure restored regional cerebral blood flow to baseline values. CONCLUSIONS: Application of high PEEP does not impair intracranial pressure or regional cerebral blood flow per se but may indirectly affect cerebral perfusion via its negative effect on macrohemodynamic variables in case of a disturbed cerebrovascular autoregulation. Therefore, following severe subarachnoid hemorrhage, a PEEP-induced decrease of mean arterial pressure should be reversed to maintain cerebral perfusion. PMID- 16215395 TI - Prevention and control of antimicrobial-resistant infections in intensive care patients. AB - OBJECTIVE: To review the literature summarizing important aspects of infection control in the critical care setting and to provide recommendations to reduce infections with resistant bacteria in the intensive care unit. DATA SOURCE: Computer searches of MEDLINE, EMBASE, and the Cochrane Library. DATA: The frequency of antibiotic-resistant, health care-associated infections has increased every year for the past 2 decades. Infections with antibiotic-resistant organisms have been linked to increases in morbidity, length of hospitalization, increased healthcare costs, and increased mortality. A comprehensive approach is necessary to prevent antimicrobial resistance in ICUs. This includes (1) preventing infections; (2) diagnosing and treating infections appropriately; (3) using antimicrobials wisely; and (4) preventing transmission. CONCLUSIONS: The reservoirs for antibiotic-resistant organisms are colonized patients, and the vectors are often healthcare workers. This places an enormous responsibility on healthcare providers to protect their patients. Clinicians must recognize the importance of adhering to the recommendations in the Centers for Disease Control's Campaign to Prevent Antimicrobial Resistance in the healthcare setting. PMID- 16215396 TI - The use of topical nonabsorbable gastrointestinal antifungal prophylaxis to prevent fungal infections in critically ill immunocompetent patients: a meta analysis. AB - OBJECTIVE: To investigate the preventive effect of topical nonabsorbable gastrointestinal antifungal prophylaxis on the incidence of fungal infection in critically ill immunocompetent patients. DATA SOURCE: Randomized controlled studies involving critically ill pediatric and adult patients in different languages from the Cochrane Controlled Trial Register (2004, issue 1), EMBASE, and MEDLINE databases (1966 to 30 April 2004) were included. Studies evaluating absorbable antifungal prophylaxis were excluded. Two reviewers assessed the quality of the studies and performed data extraction independently. DATA: Amphotericin B and nystatin were used as the nonabsorbable antifungal prophylaxis in the 15 studies included in this meta-analysis. Ten studies used a concomitant systemic antibiotic and four more studies used concomitant topical nonabsorbable antibiotics in the treatment group. Only one study compared topical nonabsorbable antifungal prophylaxis alone with placebo. The total incidence of fungal infections (relative risk [RR], 0.30; 95% confidence interval [CI], 0.18-0.48; p < .00001; extent of inconsistency [I(2)] = 0%) and proportion of patients with fungal infection (RR, 0.50; 95% CI, 0.28-0.87; p = .02; I(2) = 0%) were significantly reduced with topical nonabsorbable antifungal prophylaxis. The incidence of fungal urinary tract infection was significantly reduced (RR, 0.27; 95% CI, 0.10-0.74; p = .01; I(2)= 0%) but not fungal pneumonia (RR, 0.57; 95% CI, 0.28-1.16; p = .12; I(2)= 0%). Fungemia and catheter-related fungal sepsis were rare and not significantly reduced with nonabsorbable antifungal prophylaxis. The results remained unchanged in the sensitivity analyses after exclusion of studies with unclear study quality or exclusion of the contribution of fungal urinary tract infections to the total incidence of fungal infections. CONCLUSIONS: In critically ill immunocompetent patients who are at high risk of fungal infection, topical nonabsorbable gastrointestinal antifungal prophylaxis was associated with a reduced incidence of urinary fungal infections and a trend toward reduction in respiratory fungal infections and fungemia. Limitations in study data are such that many of these infections may have represented superficial infections of uncertain clinical importance; a large, randomized, controlled trial is needed to assess the cost-effectiveness and safety of topical nonabsorbable antifungal prophylaxis in critically ill patients. PMID- 16215397 TI - Augmentation of hospital critical care capacity after bioterrorist attacks or epidemics: recommendations of the Working Group on Emergency Mass Critical Care. AB - The Working Group on Emergency Mass Critical Care was convened by the Center for Biosecurity of the University of Pittsburgh Medical Center and the Society of Critical Care Medicine to provide recommendations to hospital and clinical leaders regarding the delivery of critical care services in the wake of a bioterrorist attack resulting in hundreds or thousands of critically ill patients. In these conditions, traditional hospital and clinical care standards in general, and critical care standards in particular, likely could no longer be maintained, and clinical guidelines for U.S. hospitals facing these situations have not been developed. The Working Group offers recommendations for this situation. PMID- 16215398 TI - Evaluation of the -26G>A CC16 polymorphism in acute respiratory distress syndrome. AB - OBJECTIVE: Different risk factors are presumably involved in the pathogenesis of acute respiratory distress syndrome (ARDS) including genetic factors. Clara cell protein 16 (CC16) is a potential candidate gene for ARDS susceptibility because reduced levels of the anti-inflammatory CC16 have been observed in bronchoalveolar lavage fluids or serum of patients with different inflammatory lung diseases. Furthermore, CC16 potently inhibits phospholipase A2, which plays a major role in ARDS pathophysiology. A functional polymorphism (-26G>A) was previously identified and related to decreased CC16 levels, asthma, and asthma severity. DESIGN: Observational study. SETTINGS: Adults with ARDS were recruited from intensive care units in two university medical centers. SUBJECTS: We evaluated the role of this genetic variant in 117 German patients with ARDS and 373 German controls. MEASUREMENTS: The CC16 -26G>A polymorphism was analyzed by melting-curve analysis using a pair of fluorescence resonance energy transfer probes. MAIN RESULTS: CC16 genotype frequencies in ARDS patients did not differ from those seen in controls. Also, the allele frequencies were identical in patients compared with controls (0.66 and 0.34). Moreover, only one of the patients who died (n = 27) was homozygous for the -26A allele. CONCLUSIONS: The CC16 -26G>A polymorphism does not affect the susceptibility to and the outcome of ARDS. PMID- 16215399 TI - Prone position, high-frequency oscillatory ventilation, and Hippocrates in acute respiratory distress syndrome. PMID- 16215400 TI - Predicting mortality: how fast can you go? PMID- 16215401 TI - Multifactorial aspects to explain variations of control group mortality rates of randomized controlled trials. PMID- 16215402 TI - The early bird catches the worm. PMID- 16215403 TI - Errors in monitoring transcutaneous PCO2 on the ear. PMID- 16215404 TI - Evolving paradigms in the management of severe traumatic brain injury. PMID- 16215405 TI - Settling the score for disseminated intravascular coagulation. PMID- 16215406 TI - The imperfect diagnosis of acute respiratory distress syndrome. PMID- 16215407 TI - Lactic acidosis in sepsis: another commentary. PMID- 16215408 TI - The unpretentious role of 2,3-diphosphoglycerate in critical illness. PMID- 16215409 TI - When shall the lung be opened up: during or after cardiac surgery? PMID- 16215410 TI - Reassessing recombinant human activated protein C for sepsis: time for a new randomized controlled trial. PMID- 16215412 TI - Exogenous surfactant in acute lung injury: no longer a question? PMID- 16215411 TI - Drotrecogin alfa (activated) treatment in severe sepsis: a "journal club" review of the global ENHANCE trial. PMID- 16215413 TI - Beta-blockade during sepsis: inspired or insane? PMID- 16215414 TI - Cardiac high-risk patients: from "permissive" to "deliberate" anemia. PMID- 16215415 TI - Peroxisome proliferator-activated receptor-gamma ligands protect against lung injury: potential therapeutic targets? PMID- 16215416 TI - The dynamic interface between hemodynamic variables and autonomic tone. PMID- 16215417 TI - Collateral damage: sepsis-induced gut injury. PMID- 16215418 TI - An old debate joined again: should we give steroids to all head-injured patients? PMID- 16215419 TI - Aneurysmal subarachnoid hemorrhage and positive end-expiratory pressure. PMID- 16215420 TI - Genetic polymorphisms in acute respiratory distress syndrome: new approach to an old problem. PMID- 16215421 TI - Controlling antibiotic-resistant bacteria: what's an intensivist to do? PMID- 16215423 TI - Prone to survive. PMID- 16215425 TI - Partial splenic embolization as pretreatment for antiviral therapy in hepatitis C virus infection. AB - Chronic hepatitis C virus infection is frequently complicated by cirrhosis and hypersplenism, which together with several other factors, such as reduced thrombopoietin synthesis in the liver, cause cytopenia. The antiviral combination therapy with pegylated interferon and ribavirin itself is impaired by haematological toxicity. Partial splenic embolization (PSE) by the injection of microspheres via a catheter comprising approximately 30-70% of the splenic parenchyma is now a safe method, which significantly reduces the cytopenia induced by hypersplenism, especially thrombocytopenia. The effect is long lasting up to 20 years and has been documented in a variety of disorders. PSE is now carefully described in a combination modality as a pretreatment to reduce cytopenia in hepatitis C virus-induced cirrhosis patients with hypersplenism, making antiviral therapy possible per se at higher dosages with a sustained duration. PMID- 16215426 TI - Safe use of pegylated interferon/ribavirin in hepatitis C virus cirrhotic patients with hypersplenism after partial splenic embolization. AB - BACKGROUND AND AIMS: Partial splenic embolization (PSE) is a non-surgical alternative for the treatment of hypersplenism. Thrombocytopenia precludes the use of pegylated interferon (peg-IFN) and ribavirin in cirrhotic patients with hepatitis C virus (HCV). We aimed to evaluate the role of PSE as a procedure allowing combined HCV therapy in this setting. METHODS: A retrospective analysis of the safety and rate of sustained virological response (SVR) after a full-dose course of peg-IFN plus ribavirin in eight HCV cirrhotic patients with severe hypersplenism undergoing PSE at a tertiary centre in Madrid, Spain, from May 2002 to August 2004. RESULTS: Six patients (75%) were in Child-Pugh class B (median score 7). PSE significantly improved the mean platelet (P = 0.012), leucocyte (P = 0.017) and haemoglobin (P = 0.035) levels, and prothrombin activity (P = 0.012). After a mean of 20 weeks after PSE all patients started weight-adjusted ribavirin plus peg-IFN-alpha2b (n = 6) or 180 microg/week of peg-IFN-alpha2a (n = 2). Six subjects (75%) completed therapy with no peg-IFN dose reductions; the dose of ribavirin was reduced in two patients reaching haemoglobin levels of less than 10 g/dl (one also received erythropoietin and granulocyte colony-stimulating factor because of neutrophil counts < 300 cells/microl). Three patients (38%) achieved SVR. Portal vein thrombosis was observed in 50% of patients, but did not preclude antiviral therapy. The pathogenic mechanism was multifactorial. It was successfully managed with anticoagulant therapy in two cases. CONCLUSIONS: PSE allowed the safe use of peg-IFN plus ribavirin in HCV cirrhotic patients with severe cytopenias who otherwise would never have been treated. The rate of SVR was 38%. PMID- 16215427 TI - Transjugular intrahepatic portosystemic stent shunt: 11 years' experience at a regional referral centre. AB - OBJECTIVES: Transjugular intrahepatic portosystemic stent shunt (TIPSS) is now widely used in the treatment of uncontrolled and recurrent variceal haemorrhage. This study reports the outcome and long-term follow-up of 125 patients who were referred to a single centre for TIPSS. METHODS: One hundred and twenty-five patients were referred to undergo TIPSS. All but 10 had variceal haemorrhage. The 10 patients referred with refractory ascites were excluded from the analysis. Our follow-up protocol was to assess shunt patency only if bleeding recurred or there was a clinical indication. The mean age was 51.5 years (range 18-87 years) and 64 patients (56%) were male. The commonest aetiology for chronic liver disease was alcohol (80%). At referral, 19 patients (16%) were Child-Pugh class A, 26 patients (23%) were Child-Pugh class B and 70 patients (61%) were Child-Pugh class C. The mean follow-up period was 20.4 months (range 0-95 months). RESULTS: TIPSS was successfully placed in 108 of 115 patients (94%). The thirty-day mortality was 30%. One-year and 2-year overall cumulative survival was 52% (survival ratio, 0.525; 95% confidence interval, 0.432-0.619) and 43% (survival ratio, 0.436; 95% confidence interval, 0.340-0.532), respectively. CONCLUSION: TIPSS is effective in the treatment of uncontrolled or recurrent variceal bleeding. In comparison with previously published studies, our study suggests no value in regular or routine shunt surveillance to reduce rebleeding episodes or mortality, but this needs to be further assessed in prospective randomized studies. PMID- 16215428 TI - Paired, quantitative measurements of hepatitis B virus DNA in saliva, urine and serum of chronic hepatitis B patients. AB - OBJECTIVES: Despite an abundance of epidemiological evidence for horizontal transmission of hepatitis B virus (HBV), the transmission route remains to be fully elucidated. In a new approach, we evaluated quantitative HBV DNA content in serum, saliva and urine as a first step in exploring possible modes of horizontal transmission. METHODS: In an outpatient setting of an academic hospital, paired serum, saliva and urine samples were collected from 150 chronically infected HBV patients. A validated HBV DNA TaqMan assay was used to quantitatively measure HBV DNA. RESULTS: Mean log HBV DNA in serum was 5.8 (range, undetectable to 10.0 log HBV DNA) copies/ml, 50% of the patients had an HBV DNA above 10 copies/ml in serum. Mean log HBV DNA level in saliva was 3.2 (range, undetectable to 7.5) copies/ml, 15% had an HBV DNA above 10 copies/ml in saliva. Mean log HBV DNA level in urine was 2.6 (range, undetectable to 5.4) copies/ml and 1% had an HBV DNA above 10 copies/ml in urine. A high, non-linear correlation was shown between HBV DNA in serum and saliva (Spearman's rho 0.82) and between serum and urine (Spearman's rho 0.74). CONCLUSIONS: The significant amounts of HBV DNA found in saliva and urine in chronic HBV patients with high viraemia in serum may have implications for the understanding of hepatitis B epidemiology. The potential infectivity of these body fluids may provide an explanation for the 20% of cases of infection obtained through horizontal transmission for which the origin of infection is yet unknown. PMID- 16215429 TI - Total body metabolism of 13C-octanoic acid is preserved in patients with non alcoholic steatohepatitis, but differs between women and men. AB - OBJECTIVES: Among numerous factors which account for the pathogenesis of non alcoholic steatohepatitis (NASH), hepatic mitochondrial beta-oxidation is considered to play a pivotal role. We performed a (13)C-based breath test with a medium-chain fatty acid to non-invasively assess total body beta-oxidation in patients with NASH and in healthy controls. METHODS: We performed a simplified (13)CO(2)-based breath test in 16 patients with histologically proven NASH and 24 healthy controls. One hundred milligrams of sodium (13)C-octanoate dissolved in 200 ml of water were orally administered and breath samples were collected before and during 3 h following administration. The samples were analysed for the cumulative (13)CO(2) recovery (%-cum-dose) by non-dispersive infrared spectrometry. Additionally, data of 69 patients who had undergone a C-octanoate breath test for the assessment of gastric emptying were retrospectively evaluated for the %-cum-dose. RESULTS: The cumulative (13)CO(2) recovery 3 h after the administration of the substrate did not differ among patients with NASH and controls (34.6 +/- 7.0% vs. 34.6 +/- 6.5%, P = 0.90). Compared with men, women yielded a significantly higher cumulative (13)CO(2) excretion in both controls (30.1 +/- 5.7% vs. 38.5 +/- 4.4%, P = 0.0008) and NASH patients (30.2 +/- 5.4% vs. 39.0 +/- 6.5%, P = 0.031). Forty-two of 69 patients (61%) of the gastric emptying group showed a normal gastric emptying rate. Among these patients, women also demonstrated a tendency for a higher (13)CO(2) recovery compared with men (P = 0.055). This was not the case in 27 patients with delayed gastric emptying (P = 0.47). CONCLUSIONS: Though hepatic mitochondrial function might be impaired in patients with NASH, total beta-oxidation of octanoic acid remains normal. Gender specific metabolic modifications seem to account for significant differences of the cumulative (13)CO(2) recovery in women and men. This may have further consequences for the appraisal of (13)C breath tests which involve octanoic acid. Further trials focusing on the assessment of body composition and energy expenditure could contribute essential further information. PMID- 16215430 TI - Hepatitis C infection: eligibility for antiviral therapies. AB - BACKGROUND: Current treatments of chronic hepatitis C virus (HCV) are effective, but expensive and susceptible to induce significant side effects. OBJECTIVES: To evaluate the proportion of HCV patients who are eligible for a treatment. METHODS: In a database comprising 1726 viraemic HCV patients, the files of 299 patients who presented to the same hepatologist for an initial appointment between 1996 and 2003 were reviewed. RESULTS: Patients' characteristics were age 43.1 +/- 15.6 years, 53% male and 92% Caucasian. The main risk factors were transfusion (43%) and drug use (22%). Genotypes were mostly genotype 1 (66%), genotype 3 (12%) and genotype 2 (10%). These characteristics were not different from those of the whole series of 1726 patients. A total of 176 patients (59%) were not treated, the reasons for non-treatment being medical contraindications (34%), non-compliance (25%) and normal transaminases (24%). In addition, 17% of patients declined therapy despite being considered as eligible, mainly due to fear of adverse events. Medical contraindications were psychiatric (27%), age (22%), end-stage liver disease (15%), willingness for pregnancy (13%), cardiac contraindication (7%) and others (16%). Only 123 patients (41%) were treated. A sustained viral response was observed in 41%. The treatment was interrupted in 16% for adverse events. CONCLUSIONS: The majority of HCV patients are not eligible for treatment. This implies that, with current therapies, only 17% of patients referred for chronic HCV become sustained responders. Some modifications of guidelines could extend the rate of treatment (patients with normal transaminases), but an important barrier remains the patients' and the doctors' fear of adverse events. PMID- 16215431 TI - Non-invasive neoplasia of the stomach. AB - Invasive gastric cancer is the final step of a cascade of genomic and phenotypic changes, which have been defined as multistep oncogenesis. This process includes a continuum of progressively dedifferentiated phenotypes, which may result in a biologically new (i.e., nuepsilonomega = neo) cell characterized by autonomous, potentially metastatic, growth (i.e., pilambdaalphazetaomega = plasia). The clinico-pathological characterization of the advanced gastric precancerous lesions has important implications for both primary and secondary cancer prevention. The WHO agency has recently redefined dysplasia as intraepithelial [i.e., non-invasive neoplasia (NiN)]; such a proposal is consistent with the biological profile of dysplasia, which shares with invasive cancer some significant molecular attributes. Long-term follow-up studies have demonstrated that, in the natural history of gastric cancer, NiN precedes invasive adenocarcinoma. In this review, the morphological features of gastric NiN are described, also illustrating differences and similarities between the current and the previously adopted histological diagnostic criteria. The molecular profile of NiN is summarized and the spectrum of the options in the clinical management of gastric NiN are reported. PMID- 16215432 TI - Risk factors for gastrointestinal malignancy in patients with iron-deficiency anaemia. AB - OBJECTIVES: Iron-deficiency anaemia (IDA) is common and may be caused by blood loss from gastrointestinal tumours. The aim of this study was to define risk factors for gastrointestinal malignancy in patients with IDA. METHODS: Patients with suspected IDA referred for gastrointestinal investigations were prospectively identified from two neighbouring UK hospitals (serving a population of 550,000 patients) between 1 January 1998 and 31 December 1999. Final diagnoses were determined after 2 years, and those patients with and without gastrointestinal cancer as a cause for their IDA were compared. Data collected included sex, age, haemoglobin, serum ferritin, mean cell volume and drug history. RESULTS: A total of 695 patients (236 men, mean age 68.5 years; 459 women, mean age 66.2 years) with IDA were investigated. Malignancy was diagnosed in 91/695 (13.1%) and gastrointestinal malignancy in 78/91 (11.2%). The most frequently diagnosed cancers were colonic (n = 44, 6.3%), gastric (n = 25, 3.6%) and renal tract (n = 7, 1%). The adjusted odds ratio (+/-95% confidence interval) for gastrointestinal cancer as a cause of IDA was significantly higher for male sex [2.96 (1.80, 4.87)], age over 50 years [7.04 (1.69, 29.32)] and haemoglobin level at presentation (< or =9.0 g/dl) [2.25 (1.29, 3.90)]. There was no significant difference in gastrointestinal malignancy in those taking aspirin (12/111, 10.8%), non-aspirin non-steroidal anti-inflammatory drugs (5/84, 6.0%) or warfarin (4/31, 12.9%) compared with those not taking these drugs (57/470, 12.1%). No cause for IDA was found in 53.7%. CONCLUSIONS: Cancer was diagnosed in 13.1% and gastrointestinal cancer in 11.2% of patients with IDA. Significant risk factors for gastrointestinal malignancy in IDA patients are male sex, age over 50 years and haemoglobin at presentation < or =9.0 g/dl. IDA should not be attributed to aspirin, non-steroidal anti-inflammatory drugs or warfarin use. PMID- 16215433 TI - Small intestinal manometry in patients with systemic sclerosis. AB - OBJECTIVES: The study explores, by the use of manometry, the frequency and severity of small intestinal involvement in patients with systemic sclerosis, and relates the manometric findings to clinical symptoms, radiology, and some intestinal regulatory peptides. METHODS: Stationary antroduodeno-jejunal manometry was used to study small bowel involvement in 10 patients with systemic sclerosis and dysmotility of the oesophagus or signs of malabsorption. Measurements were made during fasting, after a meal, and after octreotide administration and were then compared with a sex-matched control group of healthy individuals. Plasma samples were taken in order to analyse levels of motilin, peptide YY, cholecystokinin, and somatostatin. RESULTS: Manometry was abnormal, with signs of intestinal pseudo-obstruction in eight out of 10 patients. In the control group, one individual had an abnormal manometry, as a result of burst activity. The mean contractile amplitudes during fasting and periods after food, spontaneous phase III periods, and octreotide-induced activity complexes were significantly reduced in the systemic sclerosis group when compared with controls. None of the patients, including two with advanced manometric intestinal disturbances, had small intestinal dilatation when examined by radiography. The plasma peptide levels did not differ significantly between the two groups. CONCLUSIONS: In eight out of 10 patients the manometric criteria for intestinal pseudo-obstruction were fulfilled, with a motility pattern consistent with both neuropathy and myopathy. The release of motility-regulating peptides was unaffected. PMID- 16215434 TI - A candidate gene approach of immune mediators effecting the susceptibility to and severity of upper gastrointestinal tract diseases in relation to Helicobacter pylori and Epstein-Barr virus infections. AB - This review focuses on immunogenetic aspects of diseases of the upper gastrointestinal tract in which infectious agents may play a role in the aetiopathogenesis, such as Helicobacter pylori, Epstein-Barr virus (EBV) and HIV. Gastric adenocarcinoma is a common cancer all around the world, with declining incidences in Europe and high incidences in Asia and central and south America. Together with gastric atrophy and peptic ulcer disease, gastric adenocarcinoma belongs to the commonest upper gastrointestinal tract diseases. These diseases are multifactorial and factors such as smoking and dietary habits contribute to the pathogenesis. More recently, scientists have turned their eyes on the host. Functional polymorphisms in the genes regulating the host immune system may contribute to the susceptibility to and progression of disease. In multifactorial and polygenetic diseases, candidate gene studies of single nucleotide polymorphisms (SNPs) detect small to moderate relative risks. Unfortunately, only a few functional SNPs have been identified. The candidate gene approach can be seen as a useful first step in exploring causal pathways between genetic determinants and complex diseases such as those mentioned above. To date, little is known about the immunogenetics of upper gastrointestinal tract diseases. We review the literature on H. pylori, EBV and gene polymorphisms that affect key immune mediators influencing the pathogenesis of the inflammatory response, such as the genes that code for the IL-1 family, TNF-alpha, lymphotoxin alpha, and IL 10. IL-1, IL-10, lymphotoxin alpha and TNF-alpha polymorphisms increase the risk of upper gastrointestinal pathogenesis in H. pylori-infected patients, whereas IL 1 and TNF-alpha polymorphisms confer risk in EBV-infected patients. PMID- 16215435 TI - Postoperative methicillin-resistant Staphylococcus aureus enteritis following hysterectomy: a case report and review of the literature. AB - Following a hysterectomy a 43-year-old woman developed colicky abdominal pain and profuse postoperative diarrhoea. Examination was unremarkable and initial investigations revealed a normal plain abdominal X-ray initially, but later there was some small bowel dilatation and evidence of raised inflammatory markers. No cause was identified at exploratory laparotomy 2 days post operation. Flexible sigmoidoscopy was normal. The patient was empirically treated with oral vancomycin for presumed Clostridium difficile diarrhoea, although subsequent stool cultures were negative for the usual intestinal pathogens and C. difficile toxin. The diarrhoea persisted for 9 days. By day 10 stool cultures had grown methicillin-resistant Staphylococcus aureus, establishing the diagnosis. To our knowledge this is the first report of methicillin-resistant S. aureus enteritis following hysterectomy. PMID- 16215437 TI - Esophageal squamous papillomatosis. AB - Esophageal squamous papillomatosis is rare and has been associated with gastroesophageal reflux and recurrent respiratory papillomatosis. We report a case of extensive esophageal papillomatosis, no airway involvement and a slowly progressive clinical course with progressive strictures and ultimately fatal squamous cell carcinoma. In-situ hybridization performed on biopsy specimens was negative for high-risk human papilloma virus types. Due to the paucity of reported cases, little is conclusively known about the etiology, natural course and best clinical management of this disease. Human papilloma virus has been linked to some, but not all, cases, and the clinical course has been reported to vary from spontaneous regression to malignant transformation. Surveillance for malignancy by conventional endoscopic biopsies or computed tomography scan appears to have low sensitivity. This case illustrates the difficulties in clinical management and establishing a definite etiology in esophageal squamous papillomatosis. PMID- 16215436 TI - Obstructive jaundice secondary to neuroendocrine tumour in a patient with von Recklinghausen's disease. AB - Neurofibromatosis type 1 (NF1) is an autosomal dominant genetic disorder, with variable clinical manifestations and unpredictable course, associated with an increased incidence of various tumours. Plexiform neurofibromas are hallmark lesions of NF1; they are slow-growing tumours, which account for substantial morbidity, including disfigurement and functional impairment, and may even be life-threatening. Neuroendocrine tumours (NETs), a rare diverse group of neoplasms, are occasionally associated with neurofibromatosis. Pancreatic NETs are tumours with an incidence of less than 1/100 000 population/year and complex patterns of behaviour, which often need complicated strategies for optimal management. We present the case of a young adult with NF1, having a unique concurrence of plexiform neurofibroma involving the liver with an ampullary NET, and we discuss step by step the management in a specialist centre. PMID- 16215438 TI - Collagenous colitis, ulcerative colitis, coeliac disease and hyperparathyroidism in one patient: implications for the management of collagenous colitis. AB - A case of collagenous colitis, ulcerative colitis and coeliac disease all occurring in one individual is reported. In addition to causing confusion over symptoms, this observation coupled with a review of the literature raises the question of whether patients with collagenous colitis should be routinely screened for coeliac disease. Furthermore, the threshold for re-investigating for the possibility of the development of ulcerative colitis may have to be lowered, particularly if symptoms change. PMID- 16215439 TI - Onset of inflammatory bowel diseases during combined alpha-interferon and ribavirin therapy for chronic hepatitis C: report of two cases. AB - We report two patients who developed an inflammatory bowel disease (IBD) shortly after beginning combined alpha-interferon and ribavirin treatment for HCV-related chronic hepatitis. The previous clinical history was negative for IBD in both patients, who developed diarrhoea and rectal bleeding 10 days and 6 months, respectively, after the initiation of therapy. The history, therapeutic management and the possible causal relationships of these cases are discussed. PMID- 16215440 TI - Hereditary hyperferritinaemia-cataract syndrome: a challenging diagnosis for the hepatogastroenterologist. AB - Hereditary hyperferritinaemia-cataract syndrome (HHCS) is a relatively rare disorder with an autosomal dominant trait. It can be caused by various mutations within the iron responsive element (IRE) of the L-ferritin gene. These mutations result in an increased translation of L-ferritin mRNA and consequently the accumulation of L-ferritin in different fluids and tissues. HHCS patients present with an isolated hyperferritinaemia in the absence of any sign of iron overload. Early onset bilateral cataract, probably due to accumulation of ferritin crystals in the lens, is the only presenting clinical manifestation. Internists, especially gastrohepatologists, should be aware of this syndrome and differentiate it from haemochromatosis which is much more frequent, in order to avoid unnecessary imaging procedures, liver biopsies and an eventual venesection therapy, which will only lead to microcytic anaemia. In the present paper we report the first cases with HHCS diagnosed in Belgium. At diagnosis, the seven known affected members of three different families had ferritin levels between 603 and 3432 microg/l (normal < 150 microg/l), and this in combination with normal iron and transferrin values. All of them were known with early-onset bilateral cataract and our postulated diagnosis of HHCS was confirmed after genetic sequencing of the L-ferritin gene, which showed a C39U point mutation in the first family, and an A40G point mutation in the IRE loop segment in the two other families. The other investigated members of the three families had normal ferritin values, no history of early-onset cataract and genetic analyses could not reveal a mutation in the IRE of their L-ferritin gene. In current clinical practice, gastroenterologists should remember that elevated ferritin levels in the absence of documented iron overload is not haemochromatosis. PMID- 16215441 TI - Adenocarcinoma as a complication of small bowel Crohn's disease. AB - Carcinoma is a recognized but rare complication of small bowel Crohn's disease. This case series emphasizes the importance of considering this diagnosis in patients with small bowel Crohn's disease. We report three cases in which patients were treated for presumed exacerbations of Crohn's, but were subsequently found to have underlying small bowel adenocarcinoma. This case series will demonstrate the need for the surgical assessment of patients with long-standing symptomatic Crohn's disease that fails to settle with conservative management. PMID- 16215442 TI - Occurrence of hepatocellular carcinoma in an apparently 'healthy' HCV patient. PMID- 16215446 TI - Outcomes for medicaid clients with substance abuse problems before and after managed care. AB - Medicaid conversion from fee for service to managed care raised numerous questions about outcomes for substance abuse treatment clients. For example, managed care criticisms include concerns that clients will be undertreated (with too short and/or insufficiently intense services). Also of interest are potential variations in outcome for clients served by organizations with assorted financial arrangements such as for-profit status versus not-for-profit status. In addition, little information is available about the impact of state Medicaid managed care policies (including client eligibility) on treatment outcomes. Subjects of this project were Medicaid clients aged 18-64 years enrolled in the Oregon Health Plan during 1994 (before substance abuse treatment managed care, N = 1751) or 1996 1997 (after managed care, N = 14,813), who were admitted to outpatient non methadone chemical dependency treatment services. Outcome measures were retention in treatment for 90 days or more, completion of a treatment program, abstinence at discharge, and readmission to treatment. With the exception of readmission, there were no notable differences in outcomes between the fee for service era clients versus those in capitated chemical dependency treatment. There were at most minor differences among various managed care systems (such as for-profit vs not-for-profit). However, duration of Medicaid eligibility was a powerful predictor of positive outcomes. Medicaid managed care does not appear to have had an adverse impact on outcomes for clients with substance abuse problems. On the other hand, state policies influencing Medicaid enrollment may have substantial impact on chemical dependency treatment outcomes. PMID- 16215448 TI - Cost-effectiveness of bupropion, nortriptyline, and psychological intervention in smoking cessation. AB - Sustained-release bupropion and nortriptyline have been shown to be efficacious in treating cigarette smoking. Psychological intervention is also recognized as efficacious. The cost and cost-effectiveness of the 2 drug therapies have not been estimated. It was hypothesized that nortriptyline would be more cost effective than bupropion. Hypotheses were not originally proposed concerning the cost-effectiveness of psychological versus drug treatment, but the 2 were compared using exploratory analyses. This was a 3 (bupropion versus nortriptyline versus placebo) by 2 (medical management alone versus medical management plus psychological intervention) randomized trial. Participants were 220 cigarette smokers. Outcome measures were cost and cost-effectiveness computed at week 52. Nortriptyline cost less than bupropion. Nortriptyline was more cost-effective than bupropion; the difference was not statistically significant. Psychological intervention cost less than the 2 drug treatments, and was more cost-effective, but not significantly so. Prospective investigations of the cost and cost effectiveness of psychological and pharmacological intervention, using adequate sample sizes, are warranted. PMID- 16215447 TI - Alcohol consumption, risk of injury, and high-cost medical care. AB - This study examined the effect of alcohol use on the probabilities of injury, inpatient hospital stay, and emergency department visit. Data were obtained from a sample of adults (N = 1219) recruited from a Northern California county. Alcohol use measures included number of drinks, heavy drinking days, and an indicator variable for problem drinking. Models were estimated for men and women separately while controlling for confounders. Results indicate that most alcohol use measures were not significantly related to injury probability or medical care utilization. Among the exceptions, problem drinking was a significant positive predictor of any emergency department visit for both sexes. When drinkers during the past year were divided into light, moderate, and heavy drinking groups and compared to lifetime abstainers, all male drinkers had a higher probability of injury, and light and moderate female drinkers had a lower probability of an emergency department visit. PMID- 16215449 TI - The Invisible Children's Project: key ingredients of an intervention for parents with mental illness. AB - This study used a collective case study design to identify key ingredients of the Invisible Children's Project, an intervention program for families in which a parent has a mental illness. Data were obtained from interviews with parents and service providers, and from family file records. Qualitative analyses were used to generate hypotheses regarding key ingredients and targeted outcomes, and to develop a testable intervention model. Key ingredients were defined as core processes, essential services, and mediators. Strong convergence across parents and providers suggested core processes defined by family-centered, strengths based, emotionally supportive, and comprehensive approaches; essential services including family case management, 24-hour crisis services, access to flexible funds, liaison and advocacy, and mediators reflecting parent-provider trust and communication/cooperation, provider-provider trust, adoption of strengths-based approaches, development of appropriate treatment plans, parent engagement, and parent self-esteem/self-efficacy. A model of the intervention is presented, and results are discussed with respect to research and policy implications. PMID- 16215450 TI - Cost-effectiveness of public sector substance abuse treatment: comparison of a managed care approach to a traditional public sector system. AB - Costs and cost-effectiveness of public sector substance abuse treatment in 2 California counties with similar substance abuse treatment system histories are compared; one county (MidState) has adopted managed care principles. As hypothesized, MidState's costs for the index treatment episode were significantly lower than SouthState's, although unexpectedly because of lower outpatient utilization. Treatment benefits in the 7 Addiction Severity Index functional areas were examined through cost-effectiveness analyses. MidState can claim greater cost-effectiveness for its treatment dollars for significant improvement in alcohol and medical functioning (compared to unsuccessful clients and those reporting no problems). When comparing both improved clients and those maintaining no problems to unsuccessful clients, MidState is more cost-effective for improving alcohol, medical, legal, and family/social functioning; and 3 outcomes important to community stakeholders and taxpayers (legal, medical, and psychiatric functioning) are more cost-effective than alcohol, drug, and employment improvement. PMID- 16215451 TI - Sociodemographic and economic comparisons of children prescribed longer-acting versus short-acting stimulant medications for attention deficit hyperactivity disorder. AB - Little research has been conducted on sociodemographic and cost disparities regarding the use of longer-acting versus short-acting stimulants in the pediatric population. Demographic characteristics and healthcare expenditures of children taking short-acting stimulants versus longer-acting stimulants for attention deficit hyperactivity disorder (ADHD) were compared. Data from the 2000 and 2001 Medical Expenditure Panel Survey, a nationally representative household survey, were analyzed for 221 children exclusively taking short-acting stimulants and 153 children exclusively taking longer-acting stimulants. No disparities in receiving short-acting as opposed to longer-acting stimulants were found by age, gender, race/ethnicity, region of the country, or insurance status. However, children in the latter group were more likely to come from higher income backgrounds and had greater psychotropic medication costs and total healthcare expenditures. For the most part, sociodemographic disparities in medication treatment for ADHD do not appear to exist once a diagnosis has been made. PMID- 16215452 TI - Rehospitalization rates of patients with bipolar disorder discharged on a mood stabilizer versus a mood stabilizer plus an atypical or typical antipsychotic. AB - One-year rehospitalization rates of patients with bipolar disorder discharged on a mood stabilizer alone, a mood stabilizer plus a typical antipsychotic, or a mood stabilizer plus an atypical antipsychotic were examined. Time to rehospitalization was measured by using the product-limit (Kaplan-Meier) formula. Twenty-three percent of patients on a mood stabilizer alone, 27% of patients on a mood stabilizer plus a typical antipsychotic, and 25% of patients on a mood stabilizer plus an atypical antipsychotic were rehospitalized within 1 year of discharge. There were no significant differences in rehospitalization rate or time to rehospitalization between groups. One-year rehospitalization rates for patients on a mood stabilizer plus olanzapine or risperidone were 25%. The number of previous psychiatric hospitalizations contributed to the risk of readmission. Risk factors and medication costs should be considered when designing the optimal treatment plan for an individual patient. Long-term prospective studies are needed to better delineate the effectiveness of different pharmacotherapeutic regimens on the long-term treatment outcomes in patients with bipolar disorder. PMID- 16215454 TI - Spiroplasma as a candidate agent for the transmissible spongiform encephalopathies. AB - The recovery of a novel Spiroplasma sp. from brain tissues from sheep with scrapie, cervids with chronic wasting disease, and from patients with Creutzfeldt Jakob disease through passage through embryonated eggs has raised the issue of the role of Spiroplasma in the transmissible spongiform encephalopathies (TSE). In this review, we have inserted into an epidemiologic infection model evidence accumulated over the past 30 years showing involvement of Spiroplasma infection in TSE. These data support our hypothesis that a Spiroplasma sp. is the causal agent of TSE, although Koch's postulates must be fulfilled to definitively answer that question. PMID- 16215455 TI - Tau-positive fine granules in the cerebral white matter: a novel finding among the tauopathies exclusive to parkinsonism-dementia complex of Guam. AB - We examined the autopsied brains of cases of 6 types of tauopathy: parkinsonism dementia complex of Guam (PDC), corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), Pick disease, Alzheimer disease (AD), and myotonic dystrophy together with Guamanian controls. Light microscopy sections of these brains were examined using anti-tau antibodies. Tau-positive fine granules (TFGs) were globe-shaped, and 3 to 6 mum in diameter, were observed predominantly in the frontal white matter in 30 of the 35 patients with PDC. However, no TFGs were found in association with PSP, myotonic dystrophy, Pick disease, AD, or CBD. Western blot analysis of frozen brain tissue taken from the PDC cases revealed that the frontal cortex was hyperphosphorylated and contained 6 tau isoforms (3R+4R tau). However, in the present study, it was revealed that the novel TFGs in the white matter of patients with PDC was composed of 4R tau. Western blot analysis of sarkosyl-insoluble tau from the white matter of the PDC cases showed 2 major bands of 60 and 64 kDa and one minor band of 67 kDa. After dephosphorylation, these bands resolved into one major band of 4-repeat (4R) tau isoform and 3 minor bands of 3-repeat (3R) and 4R tau isoforms. Moreover, the TFGs observed in cases in which the number of neurofibrillary tangles (NFTs) was higher than the threshold level were not correlated with the presence of cortical NFTs. In conclusion, these novel TFGs were found almost exclusively in PDC brains and could therefore be considered as a characteristic neuropathologic marker of this particular tauopathy. The TFGs were hyperphosphorylated tau-positive structures that may be formed by a different mechanism from that used to produce cortical NFTs. PMID- 16215456 TI - Genotypically defined lissencephalies show distinct pathologies. AB - Lissencephaly is traditionally divided into 2 distinct pathologic forms: classic (type I) and cobblestone (type II). To date, mutations in 4 genes, LIS1, DCX, RELN, and ARX, have been associated with distinct type I lissencephaly syndromes. Each of these genes has been shown to play a role in normal cell migration, consistent with the presumed pathogenesis of type I lissencephaly. Based on these data, we hypothesized that all forms of radiographically defined type I lissencephaly independent of genotype would be pathologically similar. To test this hypothesis, we examined brains from 16 patients, including 15 lissencephalic patients and one patient with subcortical band heterotopia. Of these 16 patients, 6 had LIS1 deletions, 2 had DCX mutations, and 2 had ARX mutations. In addition, 6 patients had no defined genetic defect, although the patient with subcortical band heterotopia exhibited the same pattern of malformation expected with an XLIS mutation. In all cases, the cortex was thickened; however, the topographic distribution of the cortical pathology varied, ranging from frontal- to occipital biased pathology to diffuse involvement of the neocortex. Although brains with LIS1 deletions exhibited the classic 4-layer lissencephalic architecture, patients with DCX and ARX mutations each had unique cytoarchitectural findings distinct from LIS1. Furthermore, 2 of the 5 patients with no known genetic defect showed a fourth type of histopathology characterized by a 2-layered cortex. Interestingly, the 2 brains with the fourth type of lissencephaly showed profound brainstem and cerebellar abnormalities. In summary, we identified at least 4 distinct histopathologic subtypes of lissencephaly that stratify with the underlying genetic defect. Based on these data, a new classification for lissencephaly is proposed that incorporates both pathologic and genetic findings. PMID- 16215457 TI - The 14-3-3 protein forms a molecular complex with heat shock protein Hsp60 and cellular prion protein. AB - The 14-3-3 protein family consists of acidic 30-kDa proteins composed of 7 isoforms expressed abundantly in neurons and glial cells of the central nervous system (CNS). The 14-3-3 protein identified in the cerebrospinal fluid provides a surrogate marker for premortem diagnosis of Creutzfeldt-Jakob disease, although an active involvement of 14-3-3 in the pathogenesis of prion diseases remains unknown. By protein overlay and mass spectrometric analysis of protein extract of NTera2-derived differentiated neurons, we identified heat shock protein Hsp60 as a 14-3-3-interacting protein. The 14-3-3zeta and gamma isoforms interacted with Hsp60, suggesting that the interaction is not isoform-specific. Furthermore, the interaction was identified in SK-N-SH neuroblastoma, U-373MG astrocytoma, and HeLa cervical carcinoma cells. The cellular prion protein (PrPC) along with Hsp60 was coimmunoprecipitated with 14-3-3 in the human brain protein extract. By protein overlay, 14-3-3 interacted with both recombinant human Hsp60 and PrPC produced by Escherichia coli, indicating that the molecular interaction is phosphorylation-independent. The 14-3-3-binding domain was located in the N terminal half (NTF) of Hsp60 spanning amino acid residues 27-287 and the NTF of PrPC spanning amino acid residues 23-137. By immunostaining, the 14-3-3 protein Hsp60 and PrPC were colocalized chiefly in the mitochondria of human neuronal progenitor cells in culture, and were coexpressed most prominently in neurons and reactive astrocytes in the human brain. These observations indicate that the 14-3 3 protein forms a molecular complex with Hsp60 and PrPC in the human CNS under physiological conditions and suggest that this complex might become disintegrated in the pathologic process of prion diseases. PMID- 16215458 TI - Anaplasia is rare and does not influence prognosis in adult medulloblastoma. AB - Histopathologic grading based on increasing anaplasia predicts clinical behavior of pediatric medulloblastomas. The present study was aimed at grading 86 medulloblastomas of adult patients (aged 18 and older) by anaplasia and analyzing the predictive power. Nodularity, desmoplasia, nuclear size, nuclear pleomorphism, necrosis, and endothelial proliferations have been evaluated. Morphometric analysis of nuclear size was performed using the Eclipse Net program. Patients treated with standard postoperative radiotherapy (35 Gy to craniospinal axis and 50 Gy to posterior fossa) were considered for correlation with survival. Pathologic data and total survival were compared by Kaplan-Meier and logrank analysis. No correlation was found between total survival duration and individual pathologic features. Cooccurrence of nuclear pleomorphism, large nuclear diameter, microvascular proliferations, and necroses did not predict outcome. Severe nuclear pleomorphism was found in 4 of 86 cases; the only large cell medulloblastoma was from an 18-year-old patient. Histopathologic factors have no clinical use for stratification of patients in risk groups. The histologic spectrum of medulloblastoma in adults is different from that in children. PMID- 16215459 TI - Monomorphous angiocentric glioma: a distinctive epileptogenic neoplasm with features of infiltrating astrocytoma and ependymoma. AB - We present 8 examples of a neoplasm with features of both astrocytoma and ependymoma that may represent a distinct clinicopathologic entity. The cerebral hemispheric tumors occurred in patients that were 3, 4, 12, 14, 15, 26, 30, and 37 years of age. All presented with seizures that, with the exception of 2, began in childhood. Magnetic resonance imaging studies showed ill-defined, T2 hyperintense, generally noncontrast-enhancing lesions that, although centered on the cortex or amygdala, extended into the underlying white matter for a short distance. Histologically, the variably infiltrative tumors were distinctively angiocentric with well-developed perivascular pseudorosettes in some cases. Longitudinal and/or circumferential orientations of perivascular cells were common also. The cells were uniform in their cytologic features from case to case and were bipolar in all but one case. A glial nature was inferred from immunoreactivity for GFAP, and ependymal differentiation was suggested by positivity for EMA in three cases and ultrastructural features in one. Overall, the tumors were biologically indolent except for one that recurred and ultimately proved fatal. PMID- 16215460 TI - Cytokines and Mycobacterium leprae induce apoptosis in human Schwann cells. AB - The development of deformities during the course of leprosy disease is a major public health concern worldwide. It is possible that cytokine production and apoptosis of Schwann cells (SCs) directly affect nerve degeneration and regeneration leading to injury of the myelin sheath and axon. In the present study, the expression of TNFalpha, TGFbeta, and their receptors, in addition to cell death triggered by cytokines or whole Mycobacterium leprae were investigated in a human SC line. The results showed the presence of TNF-Rs and TGF-RII on the SC membrane and the shedding of TNF-Rs during the culture period. Evaluation of cell death was performed through TUNEL and flow cytometry techniques. TNFalpha/TGFbeta combination as well as M. leprae infection triggered an increase in the apoptosis rate in the cultured SC. Moreover, reverse transcriptase polymerase chain reaction assay revealed that M. leprae upregulated the expression of such cytokines and their receptors on the SC line. Despite the detection of TNFalpha mRNA, no protein was found in the culture supernatants. The data indicate that induction of SC death after cell interaction with M. leprae may, in fact, be implicated in the pathogenesis of nerve damage, which can most likely be modulated by in vivo cytokine production. PMID- 16215461 TI - Altered expression of immune defense genes in pilocytic astrocytomas. AB - Pilocytic astrocytoma (WHO grade I) is a circumscribed, slowly growing, benign astrocytoma that most frequently develops in the cerebellar hemispheres and in midline structures and occurs predominantly in childhood and adolescence. In contrast to diffusely infiltrating gliomas in adults (e.g. grade II astrocytomas, oligodendrogliomas), survival of patients with pilocytic astrocytoma is excellent after surgical intervention. To search for potential molecular mechanisms underlying its benign biologic behavior, we compared gene expression profiles of pilocytic astrocytomas (8 cases) with those of normal cerebellum (4 cases), low grade astrocytomas (WHO grade II; 15 cases), and oligodendrogliomas (WHO grade II; 17 cases) by cDNA array analysis. A number of immune system-related genes such as HLA-DRalpha, HLA-DPB1, HLA-DQB1, IgG3, IgGK, FCER1G, A2M, FCRN, IFI-56K, and DAP12 were upregulated in pilocytic astrocytomas relative to normal cerebellum, grade II astrocytomas, and oligodendrogliomas. Genes expressed at higher levels in pilocytic astrocytomas than in grade II astrocytomas and oligodendrogliomas include HLA-DRalpha, HLA-DPA1, HLA-DPB1, HLA-DQB1, A2M, TIMP1, TIMP2, CDKN1A, and SOCS3 and those expressed at lower levels include EGFR and PDGFRA. Hierarchical clustering analysis using the entire set of 1176 genes distinguished pilocytic astrocytomas from grade II astrocytomas and oligodendrogliomas. Clustering analysis using selected subgroups of genes based on their molecular functions revealed that immune system-related genes (75 genes) or cell adhesion, migration, and angiogenesis-related genes (69 genes) showed similar power to the entire gene set for separation of pilocytic astrocytomas from diffusely infiltrating low-grade gliomas. Immunohistochemistry revealed that HLA-DRalpha is expressed diffusely in neoplastic cells in pilocytic astrocytomas, whereas in oligodendrogliomas, expression was limited to scattered reactive astrocytes. These results suggest that gene expression profiles of pilocytic astrocytomas differ significantly from those of diffusely infiltrating low-grade gliomas and that their benign biologic behavior may be related to upregulation of immune defense-associated genes. PMID- 16215462 TI - Sporadic Creutzfeldt-Jakob disease: the extent of microglia activation is dependent on the biochemical type of PrPSc. AB - In prion-related encephalopathies, microglial activation occurs early and is dependent on accumulation of disease-specific forms of the prion protein (PrPSc) and may play a role in nerve cell death. Previously, we found that different types of PrPSc (i.e. type 1 and type 2) coexisted in approximately 25% of patients with sporadic Creutzfeldt-Jakob disease (CJD); and a close relationship was detected between PrPSc type, the pattern of PrP immunoreactivity, and extent of spongiform degeneration. To investigate whether microglial reaction is related to the biochemical type and deposition pattern of PrPSc, we carried out a neuropathologic and biochemical study on 26 patients with sporadic CJD, including all possible genotypes at codon 129 of the prion protein gene. By quantitative analysis, we demonstrated that strong microglial activation was associated with type 1 PrPSc and diffuse PrP immunoreactivity, whereas type 2 PrPSc and focal PrP deposits were accompanied by mild microglia reaction. These findings support the view that the phenotypic heterogeneity of sporadic CJD is largely determined by the physicochemical properties of distinct PrPSc conformers. PMID- 16215464 TI - Pediatric closed head injuries treated in an observation unit. AB - BACKGROUND: Closed head injury (CHI) is common in childhood and frequently results in hospital admission for observation and treatment. Observation units (OUs) have shown significant benefits for patients and physicians. At our institution, a level 1 pediatric trauma center, patients with CHI are often admitted to an OU for up to 24 hours of observation and treatment. STUDY OBJECTIVES: To describe characteristics of patients with a CHI admitted to a pediatric OU and to identify demographic, historical, clinical, and radiographic factors associated with the need for unplanned inpatient admission (UIA) after OU management. METHODS: Retrospective cohort review of all OU admissions for CHI at Primary Children's Medical Center (PCMC) from August 1999 through July 2001. Data collected included age, gender, mechanism of injury, presenting symptoms, physical examination findings, head computed tomography (CT) results, diagnosis, length of stay, outcome of the injury, and need for UIA. RESULTS: During the study period, 827 patients were seen in the ED for CHI. Two hundred eighty-five patients (34%) were admitted to the OU, 273 (33%) were admitted to an inpatient service, and 269 (33%) were discharged home. OU patients had a median age of 5.2 years, ranging from 2 weeks to 17 years. Sixty-one percent were male. The median admission length of stay was 13 hours. Common mechanisms of injury included: falls (60%), motor vehicle accidents (12%), bicycle accidents (10%), impacts from objects (9%), auto-pedestrian accidents (4.6%), and snow-related accidents (4.6%). Presenting symptoms in the ED included vomiting (39%), loss of consciousness (26%), amnesia to event (19%), persistent amnesia (5%), and seizures (4%). Physical examination findings noted in the ED included altered mental status (45%), facial abnormalities (43%), scalp abnormalities (38%), and neurologic deficits (9%). Two hundred eighty patients (98%) admitted to the OU had a head CT performed. Skull fractures were present in 109 patients (39%) and intracranial pathology (ie, epidural hematoma, subdural hematoma, or intraparenchymal contusion) was present in 38 patients (13%). Only 13 patients (5%) required admission to an inpatient service from the OU for the following reasons: continued need for intravenous (IV) fluids (n = 5), venous thrombosis (n = 2), persistent CSF leakage (n = 3), decreased level of consciousness (n = 1), pain management (n = 1), and clearing of the patient's cervical spine (n = 1). No patient deteriorated or required neurosurgery. Patients with basilar skull fractures, a head laceration (scalp or facial), and patients that needed IV fluids in the ED were more likely to need inpatient admission after a 24-hour observation stay. Logistic regression analysis identified basilar skull fractures (OR 11.61), face/scalp lacerations (OR 7.52), and the need for ED IV fluid administration (OR 4.26) to be associated with UIA. Most children with these findings were successfully discharged within 24 hours, however. Age, sex, loss of consciousness, seizure, vomiting, amnesia, altered mental status, neurologic deficits, intracranial pathology, and skull fractures (aside from basilar skull fractures) were not related to UIA. CONCLUSION: The vast majority (96%) of pediatric OU patients with CHI such as small intracranial hematomas, skull fractures, and concussions were discharged safely within 24 hours without serious complications. The presence of a basilar skull fracture, head laceration, and the need for ED IV fluids were associated with increased risk of UIA. OU admission is an efficient and effective management setting for children with stable intracranial pathology, skull fractures, and concussions. PMID- 16215463 TI - Pathology of the insular cortex in Alzheimer disease depends on cortical architecture. AB - The insular cortex plays important roles in a variety of regulatory mechanisms ranging from visceral control and sensation to covert judgments regarding inner well-being. The dementia of Alzheimer disease (AD) often includes behavioral dyscontrol and visceral dysfunction not observed in other diseases affecting cognition. This could be related to autonomic instability and to loss of the sense of self, and pathologic changes within the insula may play essential roles. The pattern of insular pathology of 17 patients with AD was examined and the severity of pathology was compared with that of the entorhinal cortex (EC), a region involved early in AD with reciprocal connections to the insula. Thioflavin S staining and Alz-50 immunostaining revealed that the insula carries a heavy burden of pathology in AD. Neurofibrillary tangles (NFTs) were largely confined to the deep layers of the cortex, whereas neuritic plaques (NPs) were distributed throughout the cellular layers and subcortical white matter. The density of NFTs, but not NPs, was highly correlated with the degree of EC pathology. However, NFTs were not seen in the insula until EC pathology reached a relatively advanced level. The density of insular NFTs varied according to architectonic type, with agranular cortex most affected, dysgranular cortex less affected, and granular cortex least affected. Thus, the insula is often involved in AD, and some of the behavioral abnormalities in AD may reflect insular pathology. PMID- 16215465 TI - Children with asthma admitted to a pediatric observation unit. AB - OBJECTIVE: Observation units (OUs) serve patients who require more evaluation or treatment than possible during an emergency department visit and who are anticipated to stay in the hospital for a short defined period. Asthma is a common admission diagnosis in a pediatric OU. Our main objective was to identify clinical factors associated with failure to discharge a child with asthma from our OU within 24 hours. METHODS: Retrospective chart review at a tertiary care children's hospital. Participants were children 2 years or older with asthma admitted from the emergency department to the OU during August 1999 to August 2001. The OU-discharged group comprised those successfully discharged from the OU within 24 hours. The unplanned inpatient admission group comprised those subsequently admitted from the OU to a traditional inpatient ward or those readmitted to the hospital within 48 hours of OU discharge. RESULTS: One hundred sixty-one children aged 2 to 20 years (median 4.0; 63% boys) met inclusion criteria; 40 patients (25%) required unplanned inpatient admission. In a multiple logistic regression model, 3 factors were associated with need for unplanned inpatient admission: female sex (adjusted odds ratio, 2.6; 95% confidence interval, 1.1-6.4; P = 0.03), temperature 38.5 degrees C or higher (adjusted odds ratio, 6.1; 95% confidence interval, 1.6-23.5; P < 0.01), and need for supplemental oxygen at the end of emergency department management (adjusted odds ratio, 5; 95% confidence interval, 1.7-15.1; P < 0.01). CONCLUSIONS: Many children with asthma can be admitted to a pediatric OU and discharged safely within 24 hours. Prospective studies are needed to confirm our findings and to identify other factors predictive of unplanned inpatient admission. PMID- 16215466 TI - Scooter injuries in children in a midwestern metropolitan area. AB - OBJECTIVES: We describe the use of nonmotorized scooters and utilization of safety equipment in a Midwestern suburb. METHODS: Schoolchildren in second to sixth grades of a local school district completed a survey distributed by school nurses and teachers in spring 2001, including topics of scooter ownership, riding patterns, and utilization of safety gear. RESULTS: Three thousand eighty-seven surveys were distributed, with a 74.4% response rate; 93.2% of children had ridden a scooter, and 71.4% owned a scooter. Children rode in many locations, with 93% riding on the sidewalk, 86.5% in the driveway, 35.5% in a parking lot, and 20% in the street. Of those owning scooters, 91.2% reported owning a helmet, 61.2% kneepads, 59.4% elbow pads, 43.8% wrist guards, and 35.5% riding gloves. Among those owning scooters, 54.6% reported wearing helmets while using their scooter, whereas 10% or less wore riding gloves, kneepads, elbow pads, or wrist guards. Helmet use decreased as grade increased; 50.7% of all second graders reported wearing helmets, compared with 22.7% of sixth graders (P < 0.001). Of those owning scooters, 32.2% reported being injured while riding their scooter. Of those injured, 54% reported wearing helmets. CONCLUSIONS: Many children ride scooters and own safety gear; however, few children wear their safety equipment when riding their scooter. Children wear helmets less frequently as they age. Physicians should continue to educate patients and their parents as to potential injuries on scooters and encourage the use of safety gear, especially helmets, when riding scooters. PMID- 16215467 TI - Mental health visits in a pediatric emergency department and their relationship to the school calendar. AB - OBJECTIVES: Over the past decade, there has been a significant increase in the rate at which children and adolescents present to emergency departments (EDs) with mental health complaints. The goal of the current study was to assess the rate of ED usage for children and adolescents and to determine if there was an association between mental health emergencies and the school year. METHODS: Retrospective chart review of 719 psychiatric consultations to an urban ED affiliated with an academic medical center. Records of consultations from April 2001 to March 2002 were reviewed and abstracted for variables such as age, sex, time and date of presentation, and psychosocial factors. RESULTS: Chi2 analyses reveal significant associations between presentation to the ED for a psychiatric complaint and time of day, day of week, and month of year. One-way analyses of variance also demonstrated mean differences in presentations for day of week and month of year. CONCLUSIONS: The current study supports previous research findings of an association between the school year and child and adolescent mental health emergencies. In the current study, the school year appears to exacerbate childhood problems, as there is a greater frequency of psychiatric emergencies while children are in school. Implications for ED program development and school based mental health service delivery are reviewed. PMID- 16215468 TI - QTc dispersion in children with severe head trauma. AB - OBJECTIVE: Corrected QT (QTc) interval prolongation has been described after subarachnoid hemorrhage and head injury in adults. Abnormal QTc prolongation is associated with a higher risk of ventricular arrhythmias. The aim of this study was to analyze QTc interval and QTc dispersion in children with severe head trauma. METHODS: Forty-three patients with severe head trauma and 49 children with no or only mild head injury as controls were enrolled in the study. QT interval from standard 12-lead electrocardiogram immediately after admission was calculated. QT interval was corrected by heart rate according to Bazett formula, and then QTc dispersion was calculated. At the same time, levels of serum electrolytes were measured. RESULTS: Although no significant difference in terms of age, sex, and R-R interval was found, QTc interval and QTc dispersion values were significantly increased in the patients with severe head trauma compared with those with no or only mild head injury (QTc, 447 +/- 31 vs. 409 +/- 27 milliseconds; QTc dispersion, 77 +/- 22 vs. 52 +/- 16 milliseconds, respectively). When the patients with severe head trauma were categorized as those with or without intracranial hemorrhage, both QTc interval and QTc dispersion were significantly greater in those with intracranial hemorrhage. These electrocardiographic parameters were inversely associated with Glasgow Coma Scale score, serum calcium levels, and, at a lesser degree, potassium levels. CONCLUSIONS: Children with severe head trauma, especially those with intracranial hemorrhage have longer QTc interval and greater QTc dispersion. PMID- 16215469 TI - Hyperventilation at referring hospitals is common before transport in intubated children with neurological diseases. AB - OBJECTIVE: To assess if cardiopulmonary complications and abnormal carbon dioxide tension are more likely in intubated children with neurological diseases undergoing transport. METHODS: We reviewed the transport records of all ventilated children retrieved to a pediatric teaching hospital in the United States within a 12-month period. RESULTS: Twenty-seven children were transported by ground (n = 11), helicopter (n = 10), and fixed-wing aircraft (n = 6). Adjustments of ventilator settings were made in 17 (63%). There were no pneumothoraces, endotracheal tube complications, arrhythmias, or cardiopulmonary resuscitation en route. Twelve patients (44%) had a primary neurological condition. In the neurological category, the pretransport blood gases revealed 7 patients with hyperventilation (Pco2, 20-29 mm Hg), and the posttransport blood gases showed 4 patients with hyperventilation (Pco2, 15-28 mm Hg). In the nonneurological category, hyperventilation occurred only in one patient before and another after transport. No significant difference between the mode of transport, stabilization time, return time, and the occurrence of hypercapnia and hypocapnia was identified. Patients who had a neurological condition were more likely to be hyperventilated at the referring hospitals (P = 0.007). Additional maneuvers were considered necessary in 3 of the 6 neurological patients and 2 of the 5 nonneurological patients with DeltapH greater than +/-0.1, whereas the management of all but one patient with DeltapH less than +/-0.1 was considered appropriate (DeltapH defined as the difference between posttransport and pretransport pH values). CONCLUSION: There is no cardiopulmonary disaster in the various modes of pediatric transport. When compared with ground transport, there is no significant increase in the risk for cardiopulmonary complications or abnormal CO2 tension in air transport of intubated children. DeltapH, in conjunction with clinical data and PCO2 values, may be a simple index for evaluation of cardiopulmonary management during transport. PMID- 16215470 TI - Esophageal coin with an unusual radiographic appearance. AB - Esophageal coins are typically seen as radiopaque disks on anteroposterior chest radiographs. This radiographic finding is generally considered classic for identifying "swallowed" coins as being in the esophagus. We report the case of an 11-year-old girl with an esophageal coin seen end-on on an anteroposterior chest radiograph and as a disk consistent with a coin in the lateral view. These findings are typically associated with coin aspiration. Clinicians should be wary of making the diagnosis of foreign body aspiration of coins by relying on single view radiographic findings alone. When clinically reasonable, we suggest additional imaging to help further localize swallowed coins. PMID- 16215471 TI - Acute myeloid leukemia presenting as spinal cord compression. AB - We report the findings of a 10-year-old boy who presented to a pediatric emergency department with symptoms of spinal cord compression. Radiological imaging demonstrated multiple soft tissue masses in the head and spine. The surgical pathology showed that the masses were comprised of myeloid leukemia cells. These findings were consistent with those observed on bone marrow aspiration. The characteristics and management of extramedullary leukemia are discussed. PMID- 16215472 TI - Small bowel obstruction caused by a congenital jejuno-jejuno band in a child. AB - A case of small bowel obstruction in a 4-year-old boy with pica is described. The child habitually chewed on his plastic toy action figures. Abdominal radiography and endoscopy confirmed the presence of chewed pieces of plastic within the digestive tract. At surgery, the cause of the obstruction was found to be an internal hernia involving a congenital fibrous band connecting 2 segments of the jejunum and unrelated to pica. Intestinal obstruction caused by foreign bodies is extremely rare, and obstruction caused by congenital bands is even more uncommon. This particular variant of a congenital band has not been previously described. Physicians should be aware that congenital bands can cause small bowel obstruction in children and that surgical treatment is required. PMID- 16215473 TI - Acute pancreatitis due to erythromycin overdose. AB - BACKGROUND: Antibiotic overdose is typically regarded as a benign event. We report a 15-year-old girl who developed pancreatitis after an overdose of erythromycin. CASE: A 15-year-old girl presented for care because of severe epigastric pain after an overdose of 5.3 g (16 x 333 mg tablets) of erythromycin base. Her physical examination was normal except for epigastric tenderness. Her serum lipase was 2024 U/L (normal, <60). She was treated with intravenous fluids and an opiate analgesic. Her serum lipase declined to 1834 and 73 U/L at 17 and 36 hours, respectively, after the initial measurement at which time she was asymptomatic. CONCLUSION: Our case supports transient pancreatitis as a potential consequence of erythromycin overdose. PMID- 16215474 TI - Tick paralysis: atypical presentation, unusual location. AB - We present 2 unrelated cases of tick paralysis presenting within a 2-month period in the greater Philadelphia region, a geographic area in which this disease is highly unusual. Our first patient demonstrated early onset of prominent bulbar palsies, an atypical presentation. Our second patient, residing in a nearby but distinct community, presented with ascending paralysis 2 months after the first. The atypical presentation of our first patient and the further occurrence within a few months of a second patient, both from the Northeastern United States where this diagnosis is rarely made, suggest the need to maintain a high index of suspicion for this disease in patients presenting with acute onset of cranial nerve dysfunction or muscle weakness. Through simple diagnostic and therapeutic measures (ie, careful physical examination to locate and remove the offending tick), misdiagnosis and unnecessary morbidity can be avoided. PMID- 16215475 TI - An adolescent scuba diver with 2 episodes of diving-related injuries requiring hyperbaric oxygen recompression therapy: a case report with medical considerations for child and adolescent scuba divers. AB - Worldwide, more than 1000 scuba (self-contained underwater breathing apparatus) diving injuries per year requiring hyperbaric recompression are documented. Approximately 80 to 90 fatalities per year are reported in North America. On average, there were 16 diving injuries requiring hyperbaric recompression therapy in scuba divers aged 19 years and younger in North America between 1988 and 2002. The youngest injured diver was 11 years old, and the youngest fatality was 14 years old during this time period. In the year 2000, certifying recreational scuba diving organizations lowered the minimum age to 8 from age 12 years for participation in the sport. We report a case of a highly trained adolescent scuba diver who, despite having advanced diving certifications, had 2 separate episodes of diving-related injuries requiring hyperbaric recompression therapy. A discussion of medical considerations in the care of the child and adolescent scuba diver is included. PMID- 16215476 TI - Playing with friends: fell on my arm. PMID- 16215477 TI - ECGs in the ED. PMID- 16215478 TI - Acute idiopathic thrombocytopenic purpura of childhood-diagnosis and therapy. PMID- 16215480 TI - Acetaminophen toxicity. PMID- 16215482 TI - Laparoscopic approach in inguinal hernia: a single technique and a tactical resource. PMID- 16215481 TI - Nucleic acid amplification tests (polymerase chain reaction, ligase chain reaction) for the diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae in pediatric emergency medicine. PMID- 16215483 TI - Laparoscopic feeding jejunostomy technique as part of staging laparoscopy. AB - Placement of a feeding jejunostomy tube is indicated for patients who need enteral access but where a gastrostomy is not feasible. This paper presents the technique and results of laparoscopic placement of feeding jejunostomy tubes in patients presenting with esophagogastric cancer. From December 2002 to February 2004, patients diagnosed with esophagogastric cancer with a potentially resectable lesion underwent staging laparoscopy. Laparoscopic feeding jejunostomy was performed on patients who were potential candidates for chemotherapy with palliative intent or neoadjuvant treatment prior to resection surgery. Surgical technique, recovery of bowel function, commencement of feeding jejunostomy, total time tube was in situ, and perioperative complications were analyzed. Of the 22 patients who underwent staging laparoscopy, a feeding jejunostomy tube was placed in 18. The remaining 4 patients were deemed to have advanced disease precluding any therapeutic options and underwent placement of esophageal stents. Feeding tubes remained in situ for a median time period of 76 days. Fourteen patients required enteral support and tubes were used for a median of 30 days. Complications from tube placement included 2 cases of wound infections, 1 of minor leak and 1 tube dislodgment. Patients were followed up for a median time of 112 days. Findings from current series suggest that placement of a feeding jejunostomy tube at the time of staging laparoscopy is a safe and reliable means of providing and maintaining nutrition for patients presenting with esophagogastric cancers. PMID- 16215484 TI - Chronic pain and recurrence after laparoscopic inguinal herniorrhaphy. AB - Chronic pain after open inguinal herniorrhaphy is a complication with an incidence of 10-20% and recurrence in 2-5% of the patients. We here present our experience with the laparoscopic technique. Patients who had undergone laparoscopic inguinal herniorrhaphy (TAPP) in our department from 1995 to 2002 received a questionnaire (N = 161). Patients were asked whether they had chronic pain or recurrence; if so, they were seen in the outpatient clinic. A total of 146 patients responded to the questionnaire (91%). Chronic pain was found in 6 patients (4%) and recurrence in 2 patients (1.4%); 134 patients (92%) had previously had inguinal herniorrhaphy with an open procedure (all Lichtenstein) on the same side. Of these, 105 (78%) preferred the laparoscopic procedure. Laparoscopic inguinal herniorrhaphy led to a very low incidence of chronic pain and recurrence compared with previous reports in open surgery. The majority of our patients were operated on for recurrent hernias, after which an even lower incidence of chronic pain and fewer recurrences in laparoscopic primary herniorrhaphies could be expected. PMID- 16215485 TI - The addition of O2 to the CO2 does not prevent the systemic effects of the CO2 pneumoperitoneum in a rabbit model. AB - CO2 pneumoperitoneum used in endoscopic surgery induces systemic effects by CO2 absorption. It was claimed that a reduction in CO2 pneumoperitoneum-induced metabolic hypoxemia was achieved by the addition of small amounts of O2 to the CO2 in a rabbit ventilated model. We reevaluated the effects of the addition of O2 to the CO2 pneumoperitoneum upon CO2 absorption in a rabbit model. The effects of a pneumoperitoneum using 100% CO2, 90% CO2 + 10% O2, 95% CO2 + 5% O2, or 100% O2 on arterial blood gases, acid base and O2 homeostasis were evaluated in nonintubated rabbits. A pneumoperitoneum pressure of 10 cm H2O (approximately 7.35 mm Hg) was used. CO2 pneumoperitoneum of 120 minutes affected blood gases and acid base homeostasis. Whereas partial pressure of CO2 and HCO3 increased (P < 0.001) during pneumoperitoneum, pH and partial pressure of O2 decreased (P < 0.001). Similar results were obtained in O2-CO2 pneumoperitoneum (P > 0.05). CO2 pneumoperitoneum profoundly affected blood gases and acid base homeostasis, resulting in metabolic hypoxemia. The addition of O2 to the CO2 did not prevent the systemic effects of CO2 pneumoperitoneum in nonintubated animals. PMID- 16215486 TI - Use of a homemade nylon loop for the prevention of postpolypectomy bleeding of large pedunculated polyps. AB - To avoid bleeding after colonoscopic polypectomy, several procedures are being used. We describe a new method that consists of a homemade nylon loop that is applied and tied to the stalk of the pedunculated polyp and after which a conventional polypectomy is done. We have used this method in 15 patients and no complication was observed. The procedure is simple; the loop may be opened to large diameters to make it easier to use for large polyps. It has a low cost and seems to be safe and effective to prevent bleeding after endoscopic polypectomy. PMID- 16215487 TI - Surgical clips found at the hepatic duct after laparoscopic cholecystectomy: a possible case of clip migration. AB - Surgical clip migration and subsequent stone formation in the common bile duct is a rare but well-established complication after laparoscopic cholecystectomy. There are some suggestions about the mechanisms of the migration process, but the details are still unclear. We report here a case in which common bile duct stones were formed around surgical clips, and other clips were found to have penetrated into the common hepatic duct, which we believe were in the process of migration after laparoscopic cholecystectomy. The patient required a laparotomy to retrieve the bile duct stones due to the distal stricture, and another laparotomy was necessary to remove the penetrating clips, which were deeply embedded in the bile duct wall. Although a variety of endoscopic and percutaneous interventional procedures are available in this era of modern medical technology, it is sometimes impractical to apply these procedures in such cases as ours, and exploratory laparotomy is sometimes required to correctly treat the patient. This case shows that the metallic surgical clips can penetrate into the intact bile duct wall through serial maceration, and we believe that careful application of clips may be the only way to prevent their migration after laparoscopic cholecystectomy. PMID- 16215488 TI - Concurrent cesarean section and hand port-assisted laparoscopic cholecystectomy: a safe approach. AB - Symptomatic gallstones are common in pregnancy. Historically, surgical management of this problem by laparoscopic cholecystectomy was deferred until after delivery and recovery. However, improved optics and tools for accessing the peritoneum make laparoscopic cholecystectomy concurrently with cesarean section an attractive option with minimal additional morbidity. We describe a new approach to laparoscopic cholecystectomy using a hand port concurrently with cesarean section. PMID- 16215489 TI - Computer-assisted laparoscopic repair of "upside-down" stomach with the Da Vinci system. AB - Recently introduced telerobotic surgical systems attempt to elude the inherent limitations of traditional laparoscopic surgery. Four patients (3 male, 1 female) with mixed hiatal and paraesophageal hernias with fixed intrathoracic partial or complete displacement of the stomach were operatively treated using the Da Vinci robotic system. Tissue dissection, hiatoplasty, and anterior hemifundoplication (Dor) were performed with the telerobotic system. There were no surgical complications. The system broke down in the fourth patient due to a software defect. Advantages were seen in terms of the intrathoracic dissection of displaced stomach through a narrow hiatus, intracorporeal suturing due to 6 degrees of freedom plus grasping. At the moment, lack of the appropriate robotic instruments for abdominal surgery as well as the enormous functional cost of the robotic system are considered to be the most significant current impediment to the adoption of robotic abdominal surgery. The continuous evolution and upgrade of the system is quite promising so far. Telerobotic-assisted hiatal hernia operation is feasible with many advantages compared with the traditional laparoscopic approach, especially during the dissection in the mediastinum in patients with intrathoracic stomach. A prospective, randomized trial will be performed later to evaluate the advantages and limitations of robotic compared with traditional laparoscopy. Technological evolution will perhaps diminish the current problems and the cost associated with robotic surgery. PMID- 16215490 TI - Combined laparoscopic near-total gastrectomy and colectomy. AB - Synchronous gastrointestinal lesions of the abdomen are uncommon. Although laparoscopic gastric resection and colonic resection have been previously described, the combined laparoscopic resection of these 2 organs is unique. We report a case of a 72-year-old man with in situ gastric cancer and high-grade dysplasia of a colon polyp who underwent successful laparoscopic resection of both lesions. This report highlights the importance of preoperative planning and port placement in combined laparoscopic procedures. PMID- 16215491 TI - Laparoscopic diagnosis and treatment of acute small bowel obstruction resulting from a congenital band. AB - Acute small bowel obstruction is a common problem, especially for those patients with previous abdominal surgery that can cause postoperative adhesions. Acute, non-postoperative small bowel obstruction is less common and has various etiologies. We report a case of acute small bowel obstruction without previous abdominal surgery. The patient underwent laparoscopic exploration, and a congenital band was found to cause direct compression of the ileum and entrapment of a segment of bowel loop. There was evidence of bowel strangulation. The color and peristalsis of the entrapped bowel loop recovered gradually after division of the band, and segmental bowel resection was avoided. He has remained asymptomatic since the procedure. We suggest early and aggressive surgical intervention for patients with acute, non-postoperative small bowel obstruction to avoid possible complications of bowel strangulation and gangrene. A laparoscopic approach may be a safe, feasible, and favorable option for correct diagnosis and appropriate treatment in this situation. PMID- 16215492 TI - Laparoscopy-assisted management of jejunal bezoar obstruction. AB - Treatment of intestinal bezoar causing obstruction is usually straightforward by an open approach, with either digital fragmentation or removal of the bezoar via an enterotomy. Herein, we report a case of small bowel bezoar obstruction treated successfully by laparoscopic technique. The bezoar was fragmented manually via a minilaparotomy and then pushed into the cecum with laparoscopic forceps. Laparoscopic management is an alternative to conventional surgery for intestinal bezoar that provides shorter hospital stay and less postoperative pain and may be recommended as the treatment of choice of such patients. PMID- 16215493 TI - Laparoscopic resection of duplicated sigmoid colon under the guidance of intraoperative colonoscopy. AB - Colonic duplications are rare congenital malformations. Complete surgical removal is the treatment of choice. The authors report a case of sigmoid colonic duplication successfully treated by laparoscopic resection under the guidance of intraoperative colonoscopy. PMID- 16215494 TI - Laparoscopic resection of presacral schwannomas. AB - Presacral tumors are particularly rare in the adult. Schwannomas are neurogenic neoplasms, rarely occurring in the retroperitoneum and the pelvis. Presented herein are the cases of 2 female patients with chronic pelvic pain who were discovered to have presacral schwannomas and were managed laparoscopically. Laparoscopy is a safe and efficient option in approaching benign pelvic tumors and might offer the advantage of better visualization of structures due to the magnification of laparoscopic view, especially in narrow anatomic spaces. PMID- 16215495 TI - Thoracoscopic repair of a right-sided congenital diaphragmatic hernia. AB - The presentation of a congenital diaphragmatic hernia (CDH) beyond infancy is rare, as it is a common cause for severe respiratory distress in the newborn. In contrast to the newborn period, delayed diagnosis of right-sided congenital diaphragmatic hernia (RCDH) is known as it causes less severe symptoms than left sided CDH. A case of an 8-month-old girl is reported. Chest x-ray and ultrasound examination of the diaphragm for symptoms of pneumonia revealed an RCDH. Thoracoscopic repair could be performed. The further course was uneventful. Thoracoscopic repair of a RCDH proved in this case to be feasible and safe beyond the newborn period. It avoided the morbidity of a major thoracotomy or laparotomy and provided an excellent cosmetic result. To our knowledge, this is the first report of thoracoscopic repair of RCDH. PMID- 16215496 TI - Case study: diagnosis and treatment of pyoderma gangrenosum. AB - Pyoderma gangrenosum, is an ulcerative, inflammatory disorder often associated in the field of stoma care with inflammatory bowel disease (IBD) (Lyon and Smith, 2001). Pyoderma gangrenosum is distinctive in that it normally has a purple rolled edge and is extremely painful. The pathogenesis is unknown but immunologic aberrations of neutrophil granulocytes seem to be important (Petering et al, 2001). This paper demonstrates the rapid development and effective multi disciplinary team management of an extensive case of Pyoderma gangrenosum. It also explores the possible causes of this particular manifestation. PMID- 16215497 TI - Colostomy irrigation: are we offering it enough? AB - This article discusses the use of irrigation for suitable colostomists and reasons why it can have a very positive effect on lifestyle. While it is evidence based it also includes anecdotal tips from patients who irrigate. The suitability of patients to irrigate and ways to 'get started' with irrigation are discussed. PMID- 16215498 TI - Hospital to home: smoothing the journey for the new ostomist. AB - For most patients, being discharged from hospital is an important milestone in their recovery. Having come through surgery, they have completed their early recovery, are well enough to leave the safe confines of the hospital and have reached the natural end point of what should be a well coordinated episode of care with a satisfactory outcome, where the discharge is as important as the admission. However, although most are naturally pleased to be going home, patients will have many fears and anxieties about how they are actually going to manage. This includes not only coping with their recovery, but also managing their lives, returning to their usual activities and lifestyle and coping with their new stoma. Knowledge information and support, nursing interventions and experiences in hospital are valuable and will be put to good use at home. This article identifies the key issues that may arise during this time, the level of knowledge and skill required for a successful transition home and the support required by the patient to adapt to their changed circumstances. PMID- 16215499 TI - Development of a nurse-led ileo-anal pouch clinic. AB - The ileo-anal pouch procedure is now seen as the operation of choice for individuals suffering with ulcerative colitis or familial adenomatous polyposis (Fazio, 1999); however, patient follow-up after this surgery remains variable. This article highlights the effectiveness of running a nurse-led follow-up service. A patient satisfaction survey has shown that having a familiar face at each follow-up visit is hugely beneficial, providing continuity and reassurance for patients and their relatives. This nurse-led clinic provides adequate time for patients to discuss a variety of issues including management of sore perianal skin and pouch dysfunction. The growth and development in clinical skills, knowledge and nursing autonomy has seen the emergence of many nurse-led clinics, where nurses are utilizing their advanced skills. Within the nurse-led ileo-anal pouch clinic in Oxford, physical and/or internal examination, and investigations, such as rigid or flexible pouchoscopy, are performed. Such procedures were positively evaluated by patients. PMID- 16215500 TI - Bridging the gap: a world-wide nursing conference. PMID- 16215502 TI - New NICE guidance for young people with cancer. PMID- 16215501 TI - Clinical treatment options for peristomal pyoderma gangrenosum. AB - Pyoderma gangrenosum is a rare debilitating skin disease of unknown aetiology that manifests itself as painful lesions and is often associated with inflammatory bowel disease. Stoma care nurses will encounter it directly around the stoma as peristomal pyoderma gangrenosum (PPG). There are many case reports of varying methods of treatment for PPG, although none have proven to be efficacious in all circumstances. When these lesions develop in the peristomal area, treatment must be balanced with the need to maintain an intact stoma appliance. PMID- 16215503 TI - Breast cancer: new treatments, new strategies. PMID- 16215504 TI - Understanding pain, part 1: physiology of pain. AB - This article is the first in a two-part series which explores pain and its management from a physiological perspective. Pain is a common experience which is often inadequately managed in hospital and community settings. While nurses are in a key position to contribute to successful pain management there are frequent accounts in the literature that nurses lack knowledge about pain management. Knowledge of pain physiology is fundamental to its successful management. This article introduces nurses to the components in the pain pathway and describes nociceptors which respond to pain stimuli and identifies pain fibres which carry pain information to the spinal cord. The processing of pain in the spinal cord is explained and the role of the brain in the manifestation of the pain experience is also explored. The gate control theory of pain and the more recent neuromatrix theory are discussed. PMID- 16215505 TI - Management and treatment options for epilepsy. AB - Epilepsy is a major chronic condition of the nervous system affecting almost 380,000 people in England and occasionally results in death of the patient. Although epilepsy and its treatment are complex, there is evidence to support a significant gap in epilepsy care, ranging from a lack of adequate record keeping of seizure occurrence to poor referral systems. This article provides a broad introduction to the condition, discusses the incidence, disease process, diagnosis, classification and the various ways, both conventional and otherwise, in which it can be managed. By understanding the above, it will assist healthcare practitioners to improve their knowledge about management of a client with epilepsy entrusted to their care. PMID- 16215507 TI - Access to medical reports. AB - This article explores what rights persons other than the patient have to access health records which come under the Data Protection Act and other statutory provisions. These other persons could include the relatives of a patient, the parents of a child and those concerned with the estate of a deceased person. In addition there are other rights of disclosure recognized by the common law and these will be considered in a later article. PMID- 16215506 TI - Registered nurse who failed in her duties to care for patients. PMID- 16215508 TI - Education in practice: a colorectal link-nurse programme. AB - This article illustrates the planning, implementation, evaluation and personal implications of a link-nurse programme for colorectal cancer nursing. The programme was designed to maintain a patient-centred support service and continue the provision of bowel cancer education and support to nurses in the clinical environment. Literature on previous link-nurse schemes, and the recent legislation on education and cancer nursing provides the background on which this programme is based. The use of educational theory supplies the theoretical underpinnings for this approach to learning, and evaluation findings are reported from both the group's and the individual's viewpoints. The programme increased the link nurses' awareness of the patient's pathway, support and information needs and the role of the multidisciplinary team in managing care. Sufficient time, support and encouragement from ward teams and clinical managers were seen as vital to the success of the programme, and specific benefits and challenges when establishing this type of practice based education for nurses are explored. PMID- 16215509 TI - Disaster preparedness and response: more than major incident initiation. AB - To ensure an efficient and effective response to disaster situations, communities must be prepared at all levels. A disaster presents problems that go beyond the scope of major incident preparedness and requires a response that utilizes all of the available resources that a community may posses, as well as assistance from outside the affected community. In order that the optimum response to a disaster situation is forthcoming, planning must take place that ensures the appropriate response across the service spectrum, including voluntary organizations. The lessons that can be learned from non-governmental organizations in developing world contexts must also be acknowledged and incorporated into the planning process. PMID- 16215510 TI - Clinical nursing rounds part 2: nurse management rounds. AB - Clinical rounds are being introduced in the Dudley Group of Hospitals to improve patient care, involve patients and improve communication. There are four types of rounds: matrons' rounds, nurse management rounds, patient comfort rounds and teaching rounds. This article describes nurse management rounds, which involve the nurse in charge of the shift seeing each patient. This gives the nurse manager an overview of the condition and needs of all the patients on the wards and the ability of staff to meet these needs. These rounds have potential benefits for patients, relatives, for the nursing team and for other healthcare professionals. PMID- 16215511 TI - A helpline for patients with chronic dermatological conditions. AB - A telephone line was set up for patients with chronic dermatological diseases in response to patients' concerns about having a point of contact for advice and for more appropriate and timely access to care when they needed to be seen. The aims were firstly to improve the quality of care and empower patients by providing advice and support with a safety net for those discharged from the authors' service. The second aim was to increase efficiency by reducing unnecessary return appointments. An audit was undertaken to evaluate the telephone service. This showed that patients were reassured that they could access the service according to their needs. There was a high level of satisfaction with the initiative and the advice received. The data also showed that there was a significant improvement in the ratio of new outpatient slots to return appointment slots and waiting times for a first appointment. PMID- 16215512 TI - Keeping nurses satisfied could be the key to keeping nurses. PMID- 16215513 TI - Embracing education in palliative care. PMID- 16215515 TI - Exploring students' experience of training for counselling skills and the impact on practice. AB - AIM: The aim of this study was to explore participants' experience of an accredited counselling skills course and their perception of its impact on practice. DESIGN: A qualitative design was used utilizing a mixed-method approach. Data were collected using semistructured interviews and the repertory grid technique. Data were analysed using thematic content analysis (interviews) and the repertory grids were analysed qualitatively using a four-stage procedure. SAMPLE: A convenience sample of six healthcare professionals undertaking a counselling skills course completed both interviews. RESULTS: The biggest impact of the course was in the area of self-awareness and skill development. The development of attentive listening, both in terms of objective and subjective findings, was evident. Students perceived that personal qualities, such as warmth and caring, were essential. All participants identified that the group were supportive and that this enhanced learning. The students changed their practice by improving their skills, increasing their awareness and by developing deeper therapeutic relationships with patients and their carers. CONCLUSION: This research adds to the body of knowledge in palliative care. It identifies key elements in the learning of counselling skills and recognizes the impact of these skills on improving the care of palliative care patients and families. PMID- 16215516 TI - Palliative care education in Croatia. PMID- 16215517 TI - Developing a nurse education network across Europe. PMID- 16215519 TI - Greater collaboration needed for education, training and equitable palliative care. PMID- 16215518 TI - Aspects of caring for dying patients which cause anxiety to first year student nurses. AB - AIM: To examine what aspects of caring for dying patients cause anxiety in student nurses during their first year of training. DESIGN: Qualitative descriptive study. SETTING: A university in the UK delivering preregistration nurse education. METHOD: A purposive sample of 38 student nurses who had just completed their first year of their programme consented to participate in the study. Data triangulation was employed, with data collected from students' reflective diaries and two focus group meetings. FINDINGS: Eight themes emerged relating to students' anxiety about caring for dying patients: coping with the physical suffering of patients: what to do or say; the severing of the relationship with the patient; the type of death; cardiopulmonary resuscitation; last offices; coping mechanisms; and interventions that would improve the student experience. CONCLUSION: Findings from the study suggest that it is aspects of the caring role, rather than personal fear of death, that form the source of much of students' anxiety. PMID- 16215520 TI - Delivering multiprofessional web-based psychosocial education: the lessons learnt. AB - Psychosocial care is one component of high-quality cancer/palliative care. The development of the internet has revolutionized the way we conduct business, shop, communicate with each other, and seek information. This article reports on the delivery of a web-based 16-week module: 'The management of cancer: psychosocial perspectives'. Twenty-nine students represented by a wide range of members of the multidisciplinary team registered for this module. Electronic-learning programmes are noted for high attrition rates, but, in this delivery, attrition rates and assessment performance were compatible with face-to-face programmes of learning. The delivery of this module indicates that there is scope for helping multiprofessional staff develop their knowledge and skills in psychosocial care. PMID- 16215521 TI - Using the arts and humanities to support learning about loss, suffering and death. AB - In this article, the benefits of using the arts and humanities as teaching and learning strategies are explored. Their contribution to undergraduate nursing curricula, as a means of promoting a deeper understanding of the experiences of loss, suffering and death, cannot be undervalued. They need to be present equally alongside the physical and social sciences in nursing curricula. More than ever, is the need for all undergraduate nursing students to participate in self reflection of these experiences, in order that they may meet the needs of their patients and families adequately, in a range of healthcare settings. PMID- 16215522 TI - Technology, futility and palliative care. PMID- 16215523 TI - Exploring district nurses' experience of a hospice at home service. AB - AIM: this study surveyed a representative sample of district nurses (DNs) to ascertain their experience of working alongside a HAH service. METHOD: a self completed postal questionnaire comprised of 14 items was distributed to 128 DNs. RESULTS: the evaluation found significant levels of satisfaction. All respondents indicated that they would refer a patient to the service again. However, the need for improved communication and increased awareness among DNs about the HAH service were also identified. CONCLUSION: the findings support the idea that a HAH service can enable patients with advanced progressive disease to be cared for at home. While the findings of this study cannot be generalized, they add to the growing body of research about HAH services in palliative care. PMID- 16215524 TI - Hospice care is a human right. PMID- 16215525 TI - Spiritual aspects of end-of-life care for Muslim patients: experiences from Iran. AB - The aim of this article is to describe the spiritual aspects of palliative care of Muslim patients based on experiences of end-of-life care in Iran. The religions of the world play a major part in the life cycle of their adherents, and most have rituals and beliefs concerning the care of dying people. For Muslims, death is believed to be not only the cessation of a complex set of biochemical processes, but also a belief that the spirit continues to live and dying is a passage from this world to the resurrection. The spirit is believed to be eternal and does not perish with death. According to Muslims' beliefs, reading of the Quran (the main religious text) can produce peace of mind in those who are near death. Nursing research has shown that the spiritual dimension of care infiltrates all aspects of nursing care. Palliative care nurses need to be informed about religious aspects of people around the world as a part of palliative care. This article indicates the methods of attending to spiritual care for Muslim patients based upon our experiences in Iran. PMID- 16215526 TI - Identifying educational needs in end-of-life care for staff and families of residents in care facilities. AB - AIM: the purpose of this article is to describe educational needs in end-of-life (EoL) care for staff and families of residents in long-term care (LTC) facilities in the province of Ontario, Canada. Barriers to providing end-of-life care education in LTC facilities are also identified. DESIGN, SETTING AND PARTICIPANTS: cross-sectional survey of directors of care in all licensed LTC facilities in the province of Ontario, Canada. RESULTS: directors of care from 426 (76.9% response rate) licensed LTC facilities completed a postal-survey questionnaire. Topics identified as very important for staff education included pain and symptom management and communication with family members about EoL care. Priorities for family education included respecting the residents' expressed wishes for care and communication about EoL care. Having sufficient institutional resources was identified as a major barrier to providing continuing education to both staff and families. CONCLUSION: through examining educational needs in EoL care this study identified an environment of inadequate staffing and over burdened care providers. The importance of increased staffing concomitant with education is a priority for LTC facilities. PMID- 16215527 TI - Corneal donation within palliative care: a review of the literature. AB - Corneal donation occurs infrequently within palliative care, even though the majority of the public say they are in favour of donating organs and tissue after death. One of the roles of the health professional is to provide patients with the necessary information to make choices, and this should include the option of donating their corneas should they so wish. This literature review explores how the subject can be raised in a systematic manner within palliative care. It looks at the potential benefits and dangers of involving the patient in such discussions, and the effects on the families. Practical issues, such as when to raise the issue, are also considered. Finally, the review discusses the implications for nursing practice and research. PMID- 16215528 TI - The staging of a hospice arts and crafts exhibition in the UK. AB - The use of arts in hospices is now well established. However, the voluntary nature of much hospice work in England encourages most regular creative activities to be provided by volunteers with minimal financial support. This article looks at how a successful arts and crafts exhibition for a hospice community was staged by a volunteer arts worker, helped by art and design students from the local university. It was not intended that the exhibition should be a research project; however, it became clear that to learn more about arts practice within a hospice environment, it would need to be appraised in some way. The project leader (and author) adopted an approach which reflected on the responses of people involved in the preparation and staging of the exhibition. PMID- 16215529 TI - An integrated model of care is needed for children and young people with cancer. PMID- 16215530 TI - [Exercise performance of patients with alveolar proteinosis]. AB - INTRODUCTION: Cardiopulmonary exercise testing (CPET) is used to evaluate the severity of interstitial lung diseases, particularly when lung function is normal. The aim of this study was to analyse exercise capacity of patients with alveolar proteinosis. METHODS: We studied 7 patients undergoing alveolar proteinosis (aged 38 +/- 5 years), Three patients complained of exertional dyspnoea, 2 had a reduced vital capacity and 5 had a DLCO of less than 75% predicted. CPET was performed on a bicycle ergometer using a standard incremental protocol. RESULTS: CPET was symptom limited for all patients.. At peak exercise, VO2 was severely reduced (19.5 +/- 5.2 ml/kg/min, 58 +/- 9%). All patients developed hyperventilation. Ventilatory reserve was 42 +/- 11% of MMV, and dead space ratio (Vd/Vt) reached 0.29 +/- 0.05. Cardio-circulatory adaptation was normal (maximum heart rate 83 +/- 9%; VO2/heart rate 70 +/- 10%). Six patients exhibited gas exchange abnormalities at peak exercise (including 4 patients having a normal vital capacity): P(A-a)O2 56 +/- 18mm Hg; PaO2 65 +/- 18 mm Hg. CONCLUSION: Patients undergoing alveolar proteinosis have severe impairment of aerobic capacity and gas exchange on exercise. CPET appears to be useful for therapeutic management. PMID- 16215531 TI - Ethics: problems of clinical conduct. AB - The study provides a framework for ethical medical conduct. A number of articles of International Law, the Italian Constitution, the Penal Code and the Medical Code of Conduct have therefore been taken into consideration. Art. 32 of the Italian Constitution, relating to the ''right to health'' is examined, paying particular attention to certain parts, and is related to Art. 35 of the European Union Charter of Fundamental Rights. In considering Art. 43 of the Penal Code, which addresses the psychological element of criminal acts, reference is made to the Medical Code of Conduct and to the Hippocratic Oath. The considerations made point up the importance of a relationship of trust and esteem between doctor and patient in clinical practice, and that this approach must be cultivated starting from the first year of university studies. PMID- 16215532 TI - Relation between length of fiber post and its mechanical retention. AB - AIM: The aim of this paper is to verify how the length of the fiber post may influence its retention in an insertion site. METHODS: Forty double conicity posts were used: 20 were inserted in epoxy resin cubes, the other 20 were inserted in extracted teeth at 2 different depth (5 mm and 10 mm). The samples were tractioned by an Instrom machine registering the postsite detach value. The root canals were examined by SEM. Statistical evaluation was made using the t test. RESULTS: The results of the sample with resin cubes show a direct proportionality between the insertion length and the post retention. This relation was not appreciated in the sample which used extracted teeth: the differences between the 5 mm and 10 mm insertion are not statistically significant. The extracted posts observation show that the fixing cement stays on the post. CONCLUSIONS: In ideal conditions the retention is directly proportional to the insertion length. The weakest interface of the adhesive system is the one between dentin and fixing cement. PMID- 16215533 TI - Clinical and therapeutic experience in twenty eight patients with burning mouth syndrome. AB - AIM: Burning mouth syndrome (BMS) is a chronic painful syndrome in clinically normal oral mucosa. There are many local, systemic and/or psychogenic pathological conditions that may clinically appear as burning and painful sensation of the oral mucosa. There are essential or idiopathic BMS and organic or secondary BMS. A diagnostic and therapeutic protocol is proposed by the authors evaluating, the effectiveness of the estroprogestinic replacement therapy in post-menopausal women suffering from idiopathic BMS. METHODS: A total of 28 patients suffering from persistent burning and painful oral sensation underwent to careful clinical and laboratory examination in order to distinguish primary forms from secondary ones. Patients suffering from primary BMS underwent to incisional biopsy for the research of estrogens receptors. RESULTS: Only 8 out of the 28 studied patients (28.5%) were negative to the clinical oral and laboratory examinations listed in our protocol and they were diagnosed for primary BMS. All of them were females. CONCLUSIONS: The use of a valid diagnostic and therapeutic protocol allows a better management of patients suffering from oral pain and a reduction if not a disappearance of the burning symptomatology in most of them. In post-menopausal patients suffering from idiopathic BMS good results may be obtained by an estroprogestinic replacement therapy. PMID- 16215534 TI - Uselessness of a questionnaire for osteoporosis and role of bone mass measurements in predicting tooth loss. AB - AIM: We evaluated whether the number of teeth lost is associated with risk factors for osteoporosis and whether bone mass measurements can add further information. METHODS: A total of 455 healthy women were enrolled. All the subjects filled in a questionnaire on risk factors for osteoporosis. The bone mineral density (BMD) was measured both by dual X-ray absorptiometry (DXA) and quantitative ultrasound measurements (QUS). RESULTS: On the basis of the questionnaire score 65.1% of the subjects were in the low risk category, 11% in the moderate risk category, 19.3% in the fairly high risk category and 4.6% in the high risk category. Close relationships (P<0.001) were observed between bone mass loss and the questionnaire risk categories. The number of teeth lost significantly increased from normal to osteoporosis groups. High correlations were also found between osteosonographic parameters and the number of teeth lost. Among questionnaire items a significant positive correlation was found only between the number of teeth lost and both age class (P<0.001) and years since menopause (P<0.001). A multiple regression showed that only age class (P<0.001) and ultrasound bone profile index (UBPI) (P=0.041) were independently linked to tooth loss. CONCLUSIONS: The results obtained showed that age is the main determinant of tooth loss and that QUS adds further information in identifying patients at a higher risk of tooth loss. PMID- 16215536 TI - Ectopic localization of Carabelli's tubercle. Review of the literature and description of two clinical cases. AB - The aim of this study is to describe 2 ectopic localizations of Carabelli's tubercle. Carabelli's tubercle or cusp is present, in a fair number of cases, in the vestibular walls of both first upper molars. Its size may vary. It is named after the Italian scientist Antonio Carabelli who first had described it in the first half of the 19(th) century. Carabelli's cusp has been often detected and studied in not hybridized ethnical groups, especially Eastern and Central Ameri can groups, in which the evaluation of heritability and sex distribution is easier. It has been found that this cusp is not clinically important, it may only sometimes interfere with orthodontic treatments. It is seldom present in the second upper molars and almost never in the vestibular walls of either side of the second upper or lower molars. After a review of the literature showing that this cusp is more rarely present in Caucasian peoples, the authors describe 2 case reports with ectopic localization. This has never been found in recent literature and therefore it is to be considered absolutely rare. PMID- 16215535 TI - Radiotherapy for maxillo-facial hemangiomas in children. Dental and periodontal long term effects. AB - Vascular maxillo-facial malformations, common pathologies in children, represent 7% of all benign tumors. They are divided into vascular malformations and hemangiomas. In the 70s-80s radiotherapy was the most applied technique, but because of its misuse or overdose, it could lead to relevant and hard consequences for the impact on the salivar glands, periodontium and growing maxillo-facial nuclei. The authors describe the case of a 38-year-old man, who, during infancy, was submitted to radiotherapy for a hemangioma to his chin and cheek. He was under observation for an agressive periodontitis, followed by a serious facial asimmetry, located in his irradiated side. They describe the therapy and the results obtained, underlying the particular difficulties detected in operating on tissues hardly and often irreversibly injured by radiotherapy. On the basis of this clinical case and of the most up-dated knowledge, therapeutical alternatives such as embolization, cryotherapy, cortisonic systemic and intralesional therapy, the use of interferon gamma 2 and the use of the most recent laser, are analyzed, and advantages and disadvantages and the most appropriate indications are underlined. PMID- 16215537 TI - Psoriasis of the dorsal surface of the tongue. AB - Psoriasis is primarily an inherited inflammatory skin disease, it is characterized by erythemato-squamous lesions that usually involve elbows, knees and the scalp. Oral manifestations are rare in psoriasis, infact, oral psoriasis involves 2% of psoriatic patients and usually it is observed with the onset of cutaneous lesions and progresses with them. Differential diagnosis should be done for Reiter's syndrome, leukoplakia and geographic tongue. The authors describe a case of tongue psoriasis without cutaneous lesions. PMID- 16215538 TI - Long-term results of Ahmed glaucoma valve and Molteno implant in neovascular glaucoma. AB - AIMS: To evaluate the surgical success results of Ahmed glaucoma valve (AGV) and Molteno single-plate implant (MSPI) in cases of neovascular glaucoma (NVG). METHODS: Between May 1997 and May 2002, 38 of 38 NVG patients that underwent implantation of AGV and 27 eyes of 27 NVG patients that underwent MSPI (a total 65 eyes of 65 patients) included to the study. RESULTS: The cumulative probabilities of success were 63.2% at 1 year, 56.2% at 2 years, 43.2% at 3 years, 37.8% at 4 years, and 25.2% at 5 years in AGV group whereas the cumulative probabilities of success were 37.0% at 1 year, 29.6% at 2 years, 29.6% at 3 years, 29.6% at 4 years, and 29.6% at 5 years in MSPI group (P=0.141). Preoperative visual acuity <2/200 (P=0.003), diagnosis of diabetes mellitius (P=0.050), and preoperative IOP>or=35 mmHg (P=0.038) were found to be poor prognostic factors for surgical success. CONCLUSIONS: Both AGV and single plate MSPI were successful for early and intermediate-term of IOP control but in long term both implants were failed to achieve control of IOP in patients with NVG. PMID- 16215539 TI - Vitreomacular traction, macular hole formation, and subfoveal choroidal neovascularization in a patient with age-related macular degeneration. PMID- 16215540 TI - South-East Scotland trabeculectomy survey. AB - AIM: Assessment of trabeculectomy in South-East Scotland (SESTS) with comparison to National Survey of Trabeculectomy (NST) and outcomes of consultant and trainee surgery. METHOD: A retrospective study of 304 consecutive, primary trabeculectomies, in two Scottish centres (Edinburgh and Dunfermline) with two glaucoma specialists and higher surgical trainees, was performed. The study group had primary open angle glaucoma (65.5%), pseudoexfoliation (6.9%), normal tension glaucoma (7.6%), pigment dispersion syndrome (1.6%), and other complicated glaucoma (18.4%). All patients underwent trabeculectomy by the glaucoma specialist (57.6%) and higher surgical trainee (42.4%) with close scrubbed supervision. RESULTS: Compared to the NST, the waiting time for surgery (<3 months) was significantly less in the SESTS (P<0.001). There was also significantly more use of intraoperative antimetabolites (P<0.01), bleb intervention (P<0.001), and a higher rate of early complications (P<0.025). There was no significant difference in outcome at intraocular pressure (IOP)<2/3 listing IOP (qualified and unqualified) between the SESTS and the NST. Significantly, more patients achieved an unqualified success of IOP<21 mmHg (P<0.01) and an unqualified success of IOP<16 mmHg in the SESTS than the NST (P<0.05). At 1 year post surgery, visual loss of greater than one Snellen line was more common in the NST (P<0.01) as was the use of anti-glaucoma medication (P<0.001). Trainee cases returned to theatre more frequently (P<0.025) and merited more bleb intervention (P<0.01) than consultant cases, but the long-term outcome was similar. CONCLUSION: Our study highlights significant changes in the practice and outcome of trabeculectomy compared to the national survey conducted a decade ago. PMID- 16215541 TI - Pattern of uveitis in a referral centre in Tunisia, North Africa. AB - AIM: To analyse the pattern of uveitis in a referral centre in Tunisia, North Africa. METHODS: The study included 472 patients with uveitis examined at the Department of Ophthalmology of Monastir (Tunisia) from January 1992 to August 2003. All patients had a comprehensive ocular and systemic history, including an extensive review of medical systems. Complete ophthalmic examination was performed in all cases, including best-corrected Snellen visual acuity, slit-lamp examination, applanation tonometry, and dilated fundus examination with three mirror lens. Standard diagnostic criteria were employed for all syndromes or entities of uveitis. RESULTS: The mean age at onset of uveitis was 34 years. The male-to-female ratio was 1:1.1. Uveitis was unilateral in 282 patients (59.7%) and bilateral in 190 patients (40.3%). Anterior uveitis was most common (166 patients; 35.2%), followed by posterior uveitis (133 patients; 28.2%), panuveitis (100 patients; 21.2%), and intermediate uveitis (73 patients; 15.5%). A specific diagnosis was found in 306 patients (64.8%). The most common cause of anterior uveitis was herpetic uveitis (56 patients; 33.7%). Toxoplasmosis was the most frequent cause of posterior uveitis (51 patients; 38.3%). Intermediate uveitis was most commonly idiopathic (63 patients; 86.3%). Behcet's disease was the most common cause of panuveitis (36 patients; 36%), followed by Vogt-Koyanagi-Harada (VKH) disease (15 patients; 15%). A total of 16 patients (3.4%) suffered from blindness, and 59 (12.5%) from uniocular blindness. CONCLUSIONS: In a hospital population in Tunisia, the most common causes of uveitis were Behcet's disease, herpes simplex infection, toxoplasmosis, and VKH disease. PMID- 16215542 TI - Comparative evaluation of diode laser trabeculoplasty vs frequency doubled Nd : YAG laser trabeculoplasty in primary open angle glaucoma. AB - AIMS: To evaluate the efficacy of diode laser trabeculoplasty (DLT) and frequency doubled Nd : YAG laser trabeculoplasty (YLT) in primary open angle glaucoma (POAG). METHODS: A total of 52 eyes of 35 patients with POAG were included. In all, 25 eyes underwent DLT (Group I) and 27 eyes underwent YLT (Group II). IOP, gonioscopy and visual fields were evaluated before and 3, 6, and 9 months and 1 year after the laser. An IOP<21 mmHg without medications or a decrease in the number of glaucoma medications with no progression of disc and field changes was considered a success. RESULTS: The mean age of patients in Group I was 57.72+/ 8.1 years and the patients in Group II had a mean age of 59.85+/-7.76 years. The mean prelaser IOP was 24.93+/-1.49 mmHg and 25.36+/-1.57 mmHg. The mean IOP after DLT at 12 months follow-up was 18.0+/-0.93 mmHg and after YLT was 18.6+/-1.52 mmHg. The reduction of IOP was statistically significant in both the groups at 3, 6, 9, and 12 months (P<0.001), however there was no significant difference between the two groups. Success rates in both groups were similar, 92% in Group I and 92.6% in Group II. CONCLUSIONS: DLT and Fd YLT are equally effective in controlling IOP in eyes with POAG. PMID- 16215544 TI - Neurotrophic keratopathy and diabetes mellitus. AB - Diabetes mellitus is frequently associated with microvascular complications such as retinopathy, nephropathy, and peripheral neuropathy. Neurotrophic keratopathy occurs in response to a neuropathy of the ophthalmic division of the trigeminal nerve. Rarely has diabetic neurotrophic keratopathy been described. This paper discusses the ophthalmic histories of three patients who presented with diabetic neurotrophic keratopathy. In one patient the corneal ulceration was the sole presenting feature of his diabetes. We discuss the need for increased vigilance in the ophthalmic community for suspecting diabetes in patients with unexplained corneal epithelial disease. PMID- 16215543 TI - Tuberculosis: an under-diagnosed aetiological agent in uveitis with an effective treatment. AB - PURPOSE: To highlight the diversity of clinical presentations with tubercular uveitis in a nonendemic setting, and discuss the diagnostic approach and an effective treatment. METHOD: Descriptive case series. RESULTS: A total of 12 cases of varied presentations of tubercular uveitis diagnosed over a period of 1 year of which six cases are described in detail. Presentations included choroidal tuberculomas, multifocal choroiditis, recurrent granulomatous uveitis, panuveitis with cystoid macular oedema, and serpiginous choroiditis. All cases had a chronic or recurrent course and responded very well to antitubercular treatment. Diagnosis was mainly assisted by positive tuberculin testing. CONCLUSION: A high index of suspicion helps diagnose ocular tuberculosis in areas of low prevalence of the disease. It forms part of the differential diagnosis of any chronic or recurrent uveitis, especially in an at-risk patient. Antitubercular treatment seems highly effective. PMID- 16215545 TI - Anterior ischaemic optic neuropathy associated with Dapsone. PMID- 16215546 TI - History of water fluoridation. PMID- 16215548 TI - Dubious discrepancies. PMID- 16215549 TI - Disability discrimination. PMID- 16215550 TI - Appreciation for Robin Cook. PMID- 16215551 TI - Career crises. PMID- 16215552 TI - Academic training model. PMID- 16215553 TI - Exhausting waste gases. PMID- 16215555 TI - Boosting dental registration for children. PMID- 16215556 TI - First for WHO Oral Health Bulletin. PMID- 16215558 TI - New organisation launched. PMID- 16215561 TI - Name change for PCDs. PMID- 16215559 TI - England's Chief Dental Officer stands down. PMID- 16215562 TI - Decontamination methods not up to scratch says study. PMID- 16215565 TI - Training set to improve dentistry in rural areas. PMID- 16215566 TI - Periodontal disease affects young adults. PMID- 16215567 TI - Oral medicine -- update for the dental practitioner: dry mouth and disorders of salivation. AB - This series provides an overview of current thinking in the more relevant areas of oral medicine for primary care practitioners, written by the authors while they were holding the Presidencies of the European Association for Oral Medicine and the British Society for Oral Medicine, respectively. A book containing additional material will be published. The series gives the detail necessary to assist the primary dental clinical team caring for patients with oral complaints that may be seen in general dental practice. Space precludes inclusion of illustrations of uncommon or rare disorders, or discussion of disorders affecting the hard tissues. Approaching the subject mainly by the symptomatic approach -- as it largely relates to the presenting complaint -- was considered to be a more helpful approach for GDPs rather than taking a diagnostic category approach. The clinical aspects of the relevant disorders are discussed, including a brief overview of the aetiology, detail on the clinical features and how the diagnosis is made. Guidance on management and when to refer is also provided, along with relevant websites which offer further detail. PMID- 16215568 TI - An unusual presentation of talon cusp: a case report. AB - The talon cusp is a relatively rare dental developmental anomaly characterised by cusp-like projections, usually observed on the lingual surface of the affected tooth. Normal enamel covers the cusp and fuses with the lingual aspect of the tooth. The cusp may or may not contain an extension of the pulp. This occurs in either maxillary or mandibular anterior teeth in both the primary and permanent dentition. This study reports the unusual case of a 47-year-old female with a taloned tooth on the right maxillary central incisor possessing both lingual and labial talons, with an x-shaped appearance when viewed occlusally. PMID- 16215569 TI - Low grade multiple myeloma that presented as a labial swelling -- a case report. AB - Myeloma is a disease of the bone marrow in which there is malignant proliferation of plasma cells. Myeloma is usually associated with the accumulation of a monoclonal immunoglobulin or light chains in plasma and subsequently an increase in light chains in the urine (Bence-Jones proteins). Renal failure can occur and bone destruction in the axial skeleton may ensue with pain and fractures. Amyloidosis associated with multiple myeloma is a relatively common finding. The most frequently reported oral sign of amyloidosis is macroglossia. There are numerous cases in the literature of amyloid deposition in tongues related to multiple myeloma. However, none of these cases describe amyloid deposition surrounding arteries in the oral cavity. We therefore report a case of an innocuous lower labial swelling, which subsequently led to the diagnosis of amyloid deposition surrounding an artery, the cause of which was later shown to be multiple myeloma. PMID- 16215576 TI - Mixed messages. PMID- 16215577 TI - Severe tissue damage. PMID- 16215578 TI - Patch test value. PMID- 16215579 TI - Hearts and mouths: perceptions of oral hygiene by at-risk heart surgery patients. AB - OBJECTIVE: To assess and use the attitudes of patients who are placed at risk after valvular heart surgery due to the connection between poor oral hygiene, valvular heart disease/surgery and the risk of developing infective endocarditis. DESIGN: A qualitative (focus group) design based study carried out on subjects three months post heart surgery. METHOD: There were five focus groups of five participants each convened by an experienced moderator. RESULTS: These portrayed an apparent pressing desire by most patients to talk about their experiences. However, patients did not accept the link between their oral health and their general health. Oral hygiene practices were not necessarily oral health related. CONCLUSIONS: The importance of the study in understanding the reasons for a patient's behaviour is evident when there is a clear need to modify the behaviour patterns of the patients effectively. Clinical trials can now be developed based on these results. PMID- 16215580 TI - General dental practitioners' views on the use of stainless steel crowns to restore primary molars. AB - AIM: To ascertain general dental practitioners' views on the use of stainless steel (pre-formed metal) crowns to restore carious primary molars. METHOD: Ninety three general dental practitioners were selected at random from those practising in Lancashire, Cheshire and Greater Manchester in 2003 and interviewed separately about the clinical care they provide to the primary dentition. Before the interview participants recorded the care they would provide for a case scenario, describing a child with a carious lesion that the British Society of Paediatric Dentistry (BSPD) guidelines indicate should be treated with a stainless steel crown. RESULTS: In answering the case scenario only six (7%) of the dentists reported that they would fit a stainless steel crown. Of the 93 dentists interviewed 71% knew of the BSPD guidelines, but only 18% had ever fitted a stainless steel crown in general practice. Reasons given for not using stainless steel crowns were they are inappropriate for many children, time consuming to fit, difficult to manipulate, expensive, and ugly. CONCLUSION: The BSPD guidelines on the use of stainless steel crowns do not reflect the views of the majority of general dental practitioners who consider these crowns unsuitable for most children and an impractical restorative technique in busy daily practice. PMID- 16215581 TI - Problems of correlations between explanatory variables in multiple regression analyses in the dental literature. AB - Multivariable analysis is a widely used statistical methodology for investigating associations amongst clinical variables. However, the problems of collinearity and multicollinearity, which can give rise to spurious results, have in the past frequently been disregarded in dental research. This article illustrates and explains the problems which may be encountered, in the hope of increasing awareness and understanding of these issues, thereby improving the quality of the statistical analyses undertaken in dental research. Three examples from different clinical dental specialties are used to demonstrate how to diagnose the problem of collinearity/multicollinearity in multiple regression analyses and to illustrate how collinearity/multicollinearity can seriously distort the model development process. Lack of awareness of these problems can give rise to misleading results and erroneous interpretations. Multivariable analysis is a useful tool for dental research, though only if its users thoroughly understand the assumptions and limitations of these methods. It would benefit evidence-based dentistry enormously if researchers were more aware of both the complexities involved in multiple regression when using these methods and of the need for expert statistical consultation in developing study design and selecting appropriate statistical methodologies. PMID- 16215593 TI - 125 years of developments in dentistry, 1880-2005. Part 2: Law and the dental profession. PMID- 16215598 TI - [Abuse of science]. PMID- 16215599 TI - [Who should be the health center physician?]. PMID- 16215600 TI - [Physicians, health and drivers licence]. PMID- 16215601 TI - [Physicians' control of driving after stroke attacks]. AB - BACKGROUND: The Finnmark Stroke Register is an investigation of new cases of stroke in a Norwegian county between April 1998 and March 2002; 622 were first attacks. The aim of this study was to estimate how physicians deal with driving issues in these patients in the sub-acute stage and during the first year after the stroke. MATERIAL AND METHODS: Evaluations of fitness to drive were made by the author and research nurses in the sub-acute stage and through telephone interviews after three and twelve months. RESULTS: 45% of first attack survivors had a valid driving licence. Usually the medical records contained no information concerning the license; this information was obtained during the follow up. 80% of patients with a licence did not fulfill the requirements for driving in the sub-acute stage because of neurological, cognitive or ophthalmologic impairment. Less than half of these patients were told not to drive. Many of the others went on driving. One year later, about 40% of patients with a valid license did not fulfill the requirements for driving. Several cases of homonymous hemianopia escaped detection for a long time. INTERPRETATION: Physicians should note the possession of a driving licence in the medical records. If indicated, acute stroke patients should receive temporary verbal and written information not to drive. Special concern ought to be given to visual disturbances. Uniform standards concerning fitness to drive ought to be developed in Norway. PMID- 16215602 TI - [No effect of supplement of essential fatty acids on lateral epicondylitis]. AB - BACKGROUND: Essential fatty acids influence the production of prostaglandins, which is suggested to be of importance for the development of chronic degenerative changes in tendons. Clinical studies indicate that treatment with essential fatty acids, vitamins and minerals may be effective against tendon diseases. This randomised trial was performed to evaluate the effect of an essential fatty acid supplement on pain in lateral epicondylitis. MATERIAL AND METHODS: 55 patients with unilateral epicondylitis were treated with eccentric training of the wrist extensor muscles for 6 months. 50% of the patients were randomised to 8 weeks of peroral supplement with essential fatty acids, 50% to placebo supplement. RESULTS: Reported pain declined throughout the study period, with a mean pain level reported on a 10 cm visual analogue scale of 4.9 cm (95% CI 4.3-5.1 cm) at inclusion and 0.95 cm (95% CI 0.5-1.1 cm) at 6 months. Maximal grip force of afflicted arm increased about 23%. No differences in pain reduction or force increase were found between the supplement and the placebo groups. CONCLUSION: Reported pain was reduced and force increased gradually over 6 months with eccentric training. No additional effect was observed with a fatty acid supplement. PMID- 16215603 TI - [Polycystic ovarian syndrome and diabetes]. AB - BACKGROUND: Polycystic ovary syndrome is the most common endocrine condition in women of fertile age. The syndrome is associated with insulin resistance, hyperinsulinaemia and diabetes. This paper reviews the association between polycystic ovary syndrome and diabetes; implications for clinical practice are suggested. MATERIAL AND METHODS: The review is based on Medline searches, our own studies, and clinical experience. RESULTS: Polycystic ovary syndrome is present in one third of women with type 1 diabetes and in almost half of all women with type 2 diabetes. In women with polycystic ovary syndrome, the prevalence of type 2 diabetes is considerably increased and gestational diabetes may occur in as many as 40%. In women with previous gestational diabetes, the risk of type 2 diabetes as well as polycystic ovarian syndrome is increased. INTERPRETATION: Women with polycystic ovary syndrome are at increased risk of developing type 2 diabetes and gestational diabetes and should be followed up accordingly. Pregnant women with polycystic ovary syndrome should have an oral glucose tolerance test as soon as the pregnancy has been confirmed. The procedure should be repeated at gestational weeks 20 and 32. Treatment with metformin should be initiated in women with type 2 diabetes who want to conceive. For the same reason metformin may also be initiated in women with type 1 diabetes. PMID- 16215604 TI - [Pathophysiologic aspects of systemic sclerosis]. AB - BACKGROUND: The pathological changes in systemic sclerosis are characterised by severe organ fibrosis, obliteration of arteries and arterioles and disturbance of immunological functions. Recent research has yielded new insight into the complex pathogenesis of systemic sclerosis, with new therapeutic options as a possible consequence. MATERIALS AND METHODS: Relevant medical literature has been reviewed. RESULTS: The pathological deposition of extracellular matrix in systemic sclerosis is most probably caused by changes in the regulation of dermal fibroblasts. The review focuses on three of the many molecules involved in the regulation of fibrosis; Transforming growth factor beta, Connective tissue growth factor and Endothelin-1. INTERPRETATION: Treatment of organ-specific disease complication has for long remained the only therapeutic option in systemic sclerosis. Based on current knowledge of the process of fibrosis, new therapeutic trials employing substances directed towards pro-fibrotic molecules are now in progress. PMID- 16215605 TI - [Treatment of Frey's syndrome using botulinum toxin]. AB - BACKGROUND: Frey's syndrome, also known as gustatory sweating, has been known for 150 years. However, effective treatment was established only as recently as in 1994, with botulinum toxin used for the treatment of torticollis or other forms of spasticity. MATERIAL AND METHODS: We present an overview over Frey's syndrome. Diagnosis with Minors sweating test and treatment with botulinum toxin is presented on the basis of our own experience. Relevant literature was found through Medline. RESULTS AND INTERPRETATION: Botulinum toxin injected intracutaneously is a safe and effective method for the treatment of Frey's syndrome. The results are convincing and the method is well documented in the literature. So far, no serious side effects have been found. PMID- 16215606 TI - [Reduced fetal movements--clinical management, recommendations and information]. AB - BACKGROUND: We reviewed the Norwegian guidelines and clinical routines in delivery units and investigated whether guidelines for management of reduced fetal movements and information given to pregnant women were based upon evidence based knowledge. MATERIAL AND METHODS: All Norwegian delivery units responded to a questionnaire about their management of pregnancies with reduced fetal movements. Literature for professionals and pregnant women was found in databases, books and journals. RESULTS: Clinical management varied significantly. All delivery units used a non-stress test when available, but few included ultrasound and Doppler examinations routinely. Five of 55 units advised women that absence of fetal movements up to 24 hours may be normal. National guidelines and university curricula recommend that distinct reduction of fetal movements require investigation. Formal kick counting was regarded as either useful, recommended or to be dissuaded. Information for pregnant women emphasises the importance of vigilance towards fetal movements, but have contradictory limits for normal fetal movements. INTERPRETATION: We have documented significant variation in clinical routines, which do not correlate with information given to pregnant women, the literature, or guidelines. This can lead to uncertainty for both pregnant women and health care professionals and may put patient safety at risk. There is a need for evidence-based guidelines. PMID- 16215607 TI - [Clinical practice variation in reduced fetal movements]. AB - BACKGROUND: Reduced fetal movements imply a risk pregnancy. This condition is present in a significant proportion of pregnancies, but both the quality of information and that of clinical care is variable. MATERIAL AND METHODS: All delivery units in Eastern Norway and Bergen have registered all consultations for reduced fetal movements, as part of the international collaborative project "Fetal Movement Intervention Assessment" (Femina). Out of 23,933 deliveries, 1200 pregnancies were examined. In 1043 pregnancies the mother had spontaneously presented her concern over reduced fetal movements; these were included in analyses. Results are presented with a 95% confidence interval. RESULTS: Asphyxia, death, growth restriction or preterm birth occurred in 19.6% (16.6 22.6%) of cases, ranging between units from 9.1% to 26.5%. Standard procedures varied extensively; ultrasonography was used in 39.0% to 98.6% and Doppler in 4.5% to 74.6% of cases. There was an association between outcomes and the procedures used. Women who waited 24 hours with reduced or absent movements before contacting healthcare had increased risk. Among those with absent movements, 47% (42-52%) had such risk behaviour. INTERPRETATION: There is a need for quality improvements in the information to pregnant women as well as in the clinical management of affected pregnancies. This could have large health benefits. We propose new guidelines for information and management. PMID- 16215608 TI - [Low-risk pregnant women in an obstetric department--how do they give birth?]. AB - BACKGROUND: The aim of the study was to investigate the obstetric outcome for low risk pregnant women delivering in a larger community hospital with 1800 deliveries annually. The investigation was carried out before the implementation of a differentiated plan for maternity care. MATERIAL AND METHODS: From January through June 2002, 920 women delivered. 520 were retrospectively considered low risk with spontaneous start of labour (56.5% of all labours). They were selected according to defined criteria. The results of this group with 197 nulliparous (37.7%) and 323 multiparous (62.1%) women are presented. RESULTS: There were 520 live newborns. 466 infants (89.6%) had an Apgar score > or = 8 after 1 minute; 509 a score > or = 8 after 5 minutes. In this low-risk group, 10.4% instrumental deliveries were performed, 2.7% caesareans and 7.7% vacuum extractions. In nulliparous women, the instrumental delivery rate was 19.7%, among the multiparous 4.6%. A surprisingly high number of low-risk women (38.7 %) were stimulated during labour with oxytocin, 62% of the primiparous and 24% of the multiparous. The indications for this were often obscure. Among oxytocin stimulated primiparous women, 31.2% had an instrumental delivery, against 1.3% in non-stimulated women. Epidural analgesia was given to 22.3% of all low-risk women; of these, 91.4% received oxytocin. 3.1% had a perineal tear grade > or = 3. INTERPRETATION: The assistance given to women in a larger community hospital is more geared towards progress of labour than that given in smaller hospitals and midwife-controlled maternity homes. Frequent use of oxytocin to enhance contractions and epidural analgesia probably increases the rate of instrumental deliveries. This investigation points to the necessity of developing differentiated plans for maternity care given to low-risk women delivering in larger community hospitals, in line with a policy adopted by the Norwegian parliament. PMID- 16215609 TI - [Continuous treatment with levodopa of Parkinson disease]. AB - BACKGROUND: The advanced stage of Parkinson's disease is characterised by motor fluctuations which are often difficult to control on traditional, peroral levodopa medication. We present our experience and a literature search regarding a method for continuous intraduodenal administration of a levodopa/carbidopa gel (Duodopa). METHODS: In a pilot study based on the compassionate use of continuous intraduodenal levodopa, patients were tested via nasoduodenal administration of the gel and on-off registration. For patients in whom a significant improvement in time in near-normal function per day was seen, permanent administration was started through a permanent duodenal port via percutaneous endoscopic gastrostomy with an inner catheter to the duodenal-jejunal transition. RESULTS AND CONCLUSION: In the nine patients tested, a significant functional improvement over time was seen. Five patients now have a permanent system with lasting good effect. Qualitative evaluation shows maintained good effect over a follow up time of up to 2.5 years (mean 19 months). We conclude that continuous enteral levodopa administration is a good and safe alternative especially for patients not offered deep brain stimulation. Its place among other treatment methods needs further assessment. PMID- 16215610 TI - [Takotsubo cardiomyopathy--potential differential diagnosis to myocardial infarction]. AB - A new syndrome, first described by the Japanese, presents as acute myocardial infarction. However, by closer examination it shows extensive akinesia in the apical and mid portions of the left ventricle, in the absence of significant coronary artery stenosis. Recently this syndrome has also been recognised outside Japan. The mechanism is unclear, but diffuse impairment of coronary microcirculation may play a role in the pathogenesis. Timely diagnosis of this potential lethal condition is mandatory, as it avoids the unnecessary risk of thrombolysis and allows adequate treatment to be started early. Diagnosis is also important for reassuring the patients that they have a good chance of complete recovery. We present three cases with this syndrome and discuss pathogenesis and clinical implications. PMID- 16215611 TI - [Use of drugs in reduced liver function]. PMID- 16215612 TI - [Percutaneous cervical cordotomy in intractable cancer pain]. AB - Percutaneous cordotomy by the lateral high cervical approach may be indicated in the treatment of intractable nociceptive or neuropathic cancer pain below the C5 dermatome. A description of the technique and two case studies are presented. PMID- 16215613 TI - [Assessment of neuropathic pain]. AB - Neuropathic pain is underdiagnosed and, in consequence, not properly treated. Neuropathic pain is defined as pain initiated or caused by a primary lesion or dysfunction in the nervous system. The term dysfunction is not clearly defined and is suggested not used. The prevalence of neuropathic pain is unknown, but a few detailed studies show that neuropathic pain is more common than previously assumed. Some common causes of neuropathic pain are entrapment/injuries to peripheral nerves, painful neuropathies, infections in the nervous system (i.e. postherpetic neuralgia), spinal cord injuries, multiple sclerosis and cerebral infarcts. Evaluation of neuropathic pain is performed on the basis of a thorough clinical examination which includes detailed questioning of different types of pain. Both spontaneous (ongoing and paroxysmal) as well as evoked pain may occur. The ongoing pain may be described by a large number of adjectives such as burning, aching, throbbing, sore, cutting, but there is no pain descriptor which is diagnostic for neuropathic pain. Pain may usually be evoked by lightly touching the skin or by exposure to cold. A clinical neurological examination is necessary in the evaluation of the level of injury, possibly also electromyography/neurography in case of peripheral nerve lesions or CT/MR in case of central lesions. Neuropathic pain is almost always characterised by changes in sensibility, usually involving the thin-fibre system (peripheral nerve fibres or central projections). Special emphasis is put on the examination of sensory dysfunction. PMID- 16215614 TI - [Genetic variation-- important for the clinical effect of opioids?]. AB - The individual variability of opioid pharmacology suggests that the patients' genetic disposition influences the response to opioids. Given the complexity of morphine pharmacology, variability may be caused by several genes. Data suggest that variability in genes encoding the enzyme metabolising morphine (UGT2B7 gene), mu-opioid receptors (OPRM1 gene) and BBB transport of morphine by multidrug resistance transporters (MDR1 gene) influence the clinical efficacy of morphine. Furthermore, variability in an enzyme-degrading catecholamines (COMT gene) may also alter the efficacy of morphine, which shows that genetic variability in non-opioid systems may indirectly influence the clinical opioid efficacy. Results obtained so far strongly suggest that opioid efficacy is partly related to inborn properties caused by genetic variability. PMID- 16215615 TI - [The role of the general practitioner in the health center]. AB - BACKGROUND: With the introduction of the list patient system, it has been suggested that routine examination of children should be transferred to the general practitioners (GPs). The aim of this study was to find out how the users of the public child health centres view the relationship between the child health centre and the GPs, and to find out whether parents would like their GPs to perform the routine examinations. MATERIAL AND METHODS: A questionnaire was given to parents in a small Norwegian town who attended the child health centre during spring and summer 2003 and to the town's GPs. Replies were received from 189 parents and from 14 out of 18 GPs. RESULTS: 29% of the parents would prefer to have their GP perform the routine examinations, 48% preferred not to and 23% did not express an opinion. Among the GPs, the corresponding figures were 57%, 29% and 14%. 93% of the GPs felt the need for closer cooperation between GP and child health centre. INTERPRETATION: The parents thought that having their GPs perform the routine examinations was less important than the contact with the health visitor and other parents. The GPs wanted closer cooperation between child health centre and GP. We recommend local trials in which the GPs perform the routine examinations of children on their list, at the child health centre. PMID- 16215618 TI - [The patent legislation in India fails the HIV/AIDS work]. PMID- 16215620 TI - [Corrupt research may damage public health]. PMID- 16215622 TI - [COX-2 inhibitors and data interpretation]. PMID- 16215625 TI - [Craving the sweet...]. PMID- 16215627 TI - [Drug addicts in physicians' offices]. PMID- 16215629 TI - Executive summary: nicotine addiction. PMID- 16215630 TI - Statin-associated neuromyotoxicity. AB - The sequelae of cardiovascular disease contribute significantly to morbidity and mortality in developed nations. As a class, the statins have been shown to measurably reduce the burden of atherosclerotic illness. However, muscle- and, more recently, nerve-related toxicity have emerged as potential complications leading to treatment withdrawal. Generally, the myopathic signs and symptoms of tenderness, myalgias, cramping and elevated serum creatine kinase (CK) activity are fully reversible after drug discontinuation. Growing evidence suggests that latent or previously minimal symptomatic muscle disease may predispose to the development of myopathy. Less information is available regarding the natural history of the sensorimotor neuropathy, but it appears to be less reversible if large fiber function is clinically manifest. Pathophysiologic clues regarding the potential causes of statin myopathy with or without neuropathy are discussed with particular attention paid to the implications of disrupted mevalonate metabolism. For example, secondary defects in isoprenoid biosynthesis are expected to impair the production of a variety of intermediaries such as dolichols, which are crucial for N-linked glycosylation; geranylgeranyl pyrophosphate, which is necessary for coenzyme Q10 and G-protein synthesis; farnesyl-pyrophosphate, which facilitates the endoproteolytic cleavage and maturation of prelamin A and modifies B-type lamins and G-proteins; and isopentenylpyrophosphate, which is involved in a nucleoside modification of selenocysteinyl-tRNA and thus indirectly related to the synthesis of all selenoproteins (estimated at 35). The nature of statin neuromyotoxicity remains unresolved; however, investigating the cellular corollaries of deranged isoprenoid metabolism may uncover clues that lead to a more complete understanding of the elusive pathophysiology. PMID- 16215631 TI - Expression and comparative analysis of genes encoding outer membrane proteins LipL21, LipL32 and OmpL1 in epidemic leptospires. AB - Leptospiral outer membrane proteins (OMPs) are highly conserved in different species, and play an essential role in the development of new immunoprotection and serodiagnosis strategies. The genes encoding LipL21, LipL32 and OmpL1 were cloned from the complete genome sequence of Leptospira interrogans serovar lai strain Lai and expressed in vitro. Sequence comparison analysis revealed that the three genes were highly conserved among distinct epidemic leptospires, including three major epidemic species Leptospira interrogans, Leptospira borgpetersenii and Leptospira weilii, in China. Immunoblot analysis was further performed to scrutinize 15 epidemic Leptospira reference strains using the antisera of the recombinant OMPs. Both immunoblot assay and reverse transcription-polymerase chain reaction demonstrated that these three OMPs were conservatively expressed in pathogenic L. interrogans strains and other pathogenic leptospires. Additionally, the use of these recombinant OMPs as antigens in enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of leptospirosis was evaluated. The recombinant LipL32 and OmpL1 proteins showed a high degree of ELISA reactivity with sera from patients infected with L. interrogans strain Lai and other pathogenic leptospires. These results may contribute to the identification of candidates for broad-range vaccines and immunodiagnostic antigens in further research. PMID- 16215632 TI - Vaccination against very virulent infectious bursal disease virus using recombinant T4 bacteriophage displaying viral protein VP2. AB - In order to develop a desirable inexpensive, effective and safe vaccine against the very virulent infectious bursal disease virus (vvIBDV), we tried to take advantage of the emerging T4 bacteriophage surface protein display system. The major immunogen protein VP2 from the vvIBDV strain HK46 was fused to the nonessential T4 phage surface capsid protein, a small outer capsid (SOC) protein, resulting in the 49 kDa SOC-VP2 fusion protein, which was verified by sodium dodecylsulfate polyacrylamide gel electrophoresis and Western blot. Immunoelectromicroscopy showed that the recombinant VP2 protein was successfully displayed on the surface of the T4 phage. The recombinant VP2 protein is antigenic and showed reactivities to various monoclonal antibodies (mAbs) against IBDV, whereas the wild-type phage T4 could not react to any mAb. In addition, the recombinant VP2 protein is immunogenic and elicited specific antibodies in immunized specific pathogen free (SPF) chickens. More significantly, immunization of SPF chickens with the recombinant T4-VP2 phage protected them from infection by the vvIBDV strain HK46. When challenged with the vvIBDV strain HK46 at a dose of 100 of 50% lethal dose (LD50) per chicken 4 weeks after the booster was given, the group vaccinated with the T4-VP2 recombinant phage showed no clinical signs of disease or death, whereas the unvaccinated group and the group vaccinated with the wild-type T4 phage exhibited 100% clinical signs of disease and bursal damages, and 30%-40% mortality. Collectively, the data herein showed that the T4 displayed VP2 protein might be an inexpensive, effective and safe vaccine candidate against vvIBDV. PMID- 16215633 TI - Differential expression of three hypoxia-inducible factor-alpha subunits in pulmonary arteries of rat with hypoxia-induced hypertension. AB - Hypoxia inducible transcription factor (HIF)-1alpha plays an important role in the development of hypoxic pulmonary hypertension, but little is known about HIF 2alpha and HIF-3alpha with respect to transcriptional regulation by hypoxia. To examine the expression patterns of all HIF-alpha subunits (HIF-1alpha, HIF-2alpha and HIF-3alpha) in pulmonary arteries of rats undergoing systemic hypoxia, five groups of healthy male Wistar rats were exposed to normoxia (N) and hypoxia for 3 (H3), 7 (H7), 14 (H14) and 21 (H21) d respectively. Mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricular hypertrophy index were measured. Lungs were inflation fixed for immunohistochemistry and in situ hybridization, and homogenized for Western blot. mPAP increased significantly after 7 d of hypoxia [(18.4+/-0.4) vs. (14.4+/-0.4) mmHg, H7 vs. N], reached its peak after 14 d of hypoxia, then remained stable. Pulmonary artery remodeling and right ventricular hypertrophy developed significantly after 14 d of hypoxia. During normoxia, HIF-1alpha and HIF-3alpha staining were slightly positive regarding mRNA levels. A substantial alteration of HIF-1alpha and HIF-3alpha staining occurred in pulmonary arteries after 14 d and 7 d of hypoxia, respectively, but HIF-2alpha staining showed an inversed trend after 14 d of hypoxia. Protein levels of all HIF-alpha subunits except HIF-3alpha showed a marked increase corresponding to the duration of hypoxia, which was obtained by Western blot. Our study found that HIF-1alpha, HIF-2alpha and HIF-3alpha may not only confer different target genes, but also play key pathogenetic roles in hypoxic-induced pulmonary hypertension. PMID- 16215634 TI - Mutational analysis of the absolutely conserved B8Gly: consequence on foldability and activity of insulin. AB - B8Gly is absolutely conserved in insulins during evolution. Moreover, its corresponding position is always occupied by a Gly residue in other members of insulin superfamily. Previous work showed that Ala replacement of B8Gly significantly decreased both the activity and the foldability of insulin. However, the effects of substitution are complicated, and different replacements sometimes cause significantly different results. To analyze the effects of B8 replacement by different amino acids, three new insulin/single-chain insulin mutants with B8Gly replaced by Ser, Thr or Leu were prepared by protein engineering, and both their foldability and activity were analyzed. In general, replacement of B8Gly by other amino acids causes significant detriment to the foldability of single-chain insulin: the conformations of the three B8 mutants are essentially different from that of wild-type molecules as revealed by circular dichroism; their disulfide stabilities in redox buffer are significantly decreased; their in vitro refolding efficiencies are decreased approximately two folds; the structural stabilities of the mutants with Ser or Thr substitution are decreased significantly, while Leu substitution has little effect as measured by equilibrium guanidine denaturation. As far as biological activity is concerned, Ser replacement of B8Gly has only a moderate effect: its insulin receptor-binding activity is 23% of native insulin. But Thr or Leu replacement produces significant detriment: the receptor-binding potencies of the two mutants are less than 0.2% of native insulin. The present results suggest that Gly is likely the only applicable natural amino acid for the B8 position of insulin where both foldability and activity are concerned. PMID- 16215635 TI - Profiling of differentially expressed genes in LRRC4 overexpressed glioblastoma cells by cDNA array. AB - Our previous study has shown that LRRC4 is a novel member of the leucine-rich repeat (LRR) superfamily and has the potential to suppress brain tumor growth. In order to further analyze the functions of LRRC4 on the maintenance of normal function and suppression of tumorigenesis in the central nervous system, we investigated alterations in gene expression related to neurobiology by the Atlas array in two inducible dual-stable LRRC4-overexpressing cell lines. Seventeen of 588 genes spotted on the Atlas membrane showed altered expression levels in LRRC4 transfected U251MG Tet-on cells, which are involved in cell proliferation and cell cycle progression, tumor invasion and metastasis, and neurotransmitter synthesis and release. In addition, cell invasion assay results showed that LRRC4 can inhibit the U251MG cell migration. These studies represent the first cDNA array analysis of the effects of LRRC4 on the involvement of different neurobiological genes in U251MG glioblastoma cells and provide new insights into the function of LRRC4 in glioma. PMID- 16215636 TI - Nitric oxide inducing function and intracellular movement of chicken interleukin 18 in cultured cells. AB - To evaluate the characteristics of chicken interleukin-18 (ChIL-18) in different forms in vitro, the ChIL-18 full-length gene (ChIL-18-F) and the ChIL-18 presumed mature protein gene (ChIL-18-M) were cloned and inserted into the eukaryotic expression vector pCI, to construct recombinant pCI-ChIL-18-F and pCI-ChIL-18-M. The recombinant plasmids were then transferred into chicken splenic lymphocytes (CSLs). Western blot showed that ChIL-18-F, with a molecular weight of 23.0 kDa, was produced in CSLs transfected by pCI-ChIL-18-F; ChIL-18-M, with a molecular weight of 19.5 kDa, was produced in CSLs transfected by pCI-ChIL-18-M. The nitric oxide (NO) level in the transfected CSLs and the culture medium at different time points was further examined under confocal microscopy using 4,5 diaminofluorescein staining. The results showed that both pCI-ChIL-18-F and pCI ChIL-18-M groups showed significant increase in intracellular and extracellular NO production compared with pCI transfected control cells. These results suggest that both ChIL-18-F and ChIL-18-M could stimulate NO secretion in CSLs. To characterize the intracellular distribution of ChIL-18, ChIL-18-F and ChIL-18-M were each fused to the enhanced green fluorescent protein gene, and expressed in Vero cells. The results showed that the ChIL-18-F tended to the membranous region in Vero cells, while ChIL-18-M did not. This indicates that the N-terminal 27 amino acid peptide helped ChIL-18 target to Vero cell membranes. PMID- 16215637 TI - Cloning, structural organization and chromosomal mapping of rat costimulatory molecule 4-1BBL. AB - 4-1BBL (TNFSF9) is a member of the tumor necrosis factor (TNF) ligand superfamily, which is expressed on some activated antigen presenting cells and B cells. We isolated a rat cDNA clone encoding the rat homologue of the human 4 1BBL (GenBank accession No. AY259541). The deduced rat 4-1BBL protein, consisting of 308 amino acids with a molecular weight of 33,469 Da, was a typical type II transmembrane glycoprotein, the same as human and murine 4-1BBL. "SDAA" in the cytoplasmic domain of rat 4-1BBL was deduced to act as the phosphorylation site for casein kinase I ("SXXS" motif), which is present in the cytoplasmic domains of human and murine 4-1BBL, and all other TNF ligand family members known to utilize reverse signaling. The two introns of 4-1BBL were also cloned (GenBank accession No. AY332409). Rat 4-1BBL is much more homologous with murine 4-1BBL than with human 4-1BBL, in both nucleotide and amino acid sequences. Rat 4-1BBL was expressed in all tested tissues: brain, lung, colon, liver, thymus, testicle, kidney, adrenal, stomach, spleen and heart. The chromosomal location of rat 4 1BBL was first identified by bioinformatics, then by fluorescence in situ hybridization at 9q11 (GenBank accession UniGene No. Rn.46783). Rat, murine and human 4-1BBL genes are evolved from a common gene. The identification and characterization of the rat counterpart of human 4-1BBL will facilitate studies of the biological function of this molecule. PMID- 16215638 TI - Purification and characterization of two endo-beta-1,4-glucanases from mollusca, Ampullaria crossean. AB - Two novel endo-beta-1,4-glucanases, EG45 and EG27, were isolated from the gastric juice of mollusca, Ampullaria crossean, by anion exchange, hydrophobic interaction, gel filtration and a second round of anion exchange chromatography. The purified proteins EG45 and EG27 appeared as a single band on sodium dodecylsulfate polyacrylamide gel electrophoresis with a molecular mass of 45 kDa and 27 kDa, respectively. The optimum pH for CMC activity was 5.5 for EG45 and 4.4-4.8 for EG27. The optimum temperature range for EG27 was broad, between 50 degrees and 60 degrees; for EG45 it was 50 degrees. The analysis on the stability of these two endo-beta-1,4-glucanases showed that EG27 was acceptably stable at pH 3.0-11.0 even when the incubation time was prolonged to 24 h at 30 degrees, whereas EG45 remained relatively stable at pH 5.0-8.0. About 85% of the activity of EG27 could be retained upon incubation at 60 degrees for 24 h. However, less than 10% residual activity of EG45 was detected at 50 degrees. Among different kinds of substrates, both enzymes showed a high preference for carboxymethyl cellulose. EG45, in particular, showed a carboxymethyl cellulose hydrolytic activity of 146.5 IU/mg protein. Both enzymes showed low activities to xylan (from oat spelt) and Sigmacell 101, and they were inactive to p-nitrophenyl-beta D-cellobioside, salicin and starch. PMID- 16215639 TI - Recombination and heterologous expression of allophycocyanin gene in the chloroplast of Chlamydomonas reinhardtii. AB - Heterogeneous expression of multiple genes in the nucleus of transgenic plants requires the introduction of an individual gene and the subsequent backcross to reconstitute multi-subunit proteins or metabolic pathways. In order to accomplish the expression of multiple genes in a single transformation event, we inserted both large and small subunits of allophycocyanin gene (apcA and apcB) into Chlamydomonas reinhardtii chloroplast expression vector, resulting in papc-S. The constructed vector was then introduced into the chloroplast of C. reinhardtii by micro-particle bombardment. Polymerase chain reaction and Southern blot analysis revealed that the two genes had integrated into the chloroplast genome. Western blot and enzyme-linked immunosorbent assay showed that the two genes from the prokaryotic cyanobacteria could be correctly expressed in the chloroplasts of C. reinhardtii. The expressed foreign protein in transformants accounted for about 2%-3% of total soluble proteins. These findings pave the way to the reconstitution of multi-subunit proteins or metabolic pathways in transgenic C. reinhardtii chloroplasts in a single transformation event. PMID- 16215640 TI - Biological activities of purified harpin(Xoo) and harpin(Xoo) detection in transgenic plants using its polyclonal antibody. AB - Many harpins have been found in plant pathogen bacteria that can elicit disease and insect resistance in plants, and promote plant growth. In this work, we overexpressed and purified Xanthomonas oryzae pv. oryzae harpin, harpin(Xoo), in Escherichia coli BL21/pGEX-hpa1. Harpin(Xoo) was fused to the C-terminus of glutathione S-transferase (GST) and purified using the Bulk GST purification module and thrombin cleavage capture kit. Purified harpin(Xoo) protein was sensitive to protease K and stable to heat treatment, and could not induce a hypersensitive response after treatment with various plant metabolic inhibitors; these characteristics were similar to harpin(Ea) of Erwinia amylovora. The purified harpin(Xoo) showed a similar ability to induce tobacco mosaic virus resistance in tobacco as harpin(Ea). Its antibody worked well in detecting the purified harpin(Xoo), harpin(Xoo) in the total protein of E. coli BL21/pGEX-hpa1 and an hpa1 transgenic rice. PMID- 16215641 TI - Haplotype analysis of NAD(P)H oxidase p22 phox polymorphisms in end-stage renal disease. AB - NAD(P)H oxidase is one of the most important sources of reactive oxygen species and has been demonstrated to be upregulated by angiotensin II in the kidney. Given the effect of angiotensin-converting enzyme inhibitors on the progression of both diabetic and non-diabetic renal disease, we hypothesized that the polymorphisms of NAD(P)H oxidase are associated with development of end-stage renal disease (ESRD). We examined five polymorphisms in the CYBA gene encoding the p22 phox component of NAD(P)H oxidase, including 242C/T and 640A/G polymorphisms in 467 ESRD patients and 490 healthy individuals. The T allele of the 242C/T polymorphism showed a protective effect against ESRD only in the nondiabetic (non-DM) group (P = 0.0095), and haplotype estimation revealed that the frequency of 242C-640A was higher in the non-DM group (46.7%) than in the control group (39.7%). The CC-AA genotype was still significantly associated with ESRD without diabetes after adjusting for confounding factors (P = 0.035). In contrast, there was no difference between the DM group and the control group. In conclusion, we identified a risk haplotype for nondiabetic ESRD in the CYBA gene using haplotype analysis. Haplotype analysis proved useful for elucidating the genetic contribution of NAD(P)H oxidase p22 phox to ESRD. PMID- 16215642 TI - Molecular genetic analysis for a novel Ael allele of the ABO blood group system. AB - The ABO blood group is the most important system in clinical transfusion medicine. Previous studies on the genetic base of the common ABO group and some rare ABO subgroups have suggested that the molecular genetic background of the ABO gene in the Chinese population has specific character. In this study, we carried out a molecular genetic analysis of a family with an individual diagnosed as Ael subgroup by serological tests. A novel allele was identified in our A subgroup cases. PMID- 16215643 TI - SYNGR1 is associated with schizophrenia and bipolar disorder in southern India. AB - Chromosome 22q11-13 is one of the most consistent linkage regions for schizophrenia (SCZ) and bipolar disorder (BPAD). The SYNGR1 gene, which is associated with presynaptic vesicles in neuronal cells, is located on 22q13.1. We have previously identified a novel nonsense mutation in the SYNGR1 gene in a SCZ pedigree. In the present study, a detailed analysis of this gene was performed in a case-control cohort (198 BPAD, 193 SCZ and 107 controls from southern India) to test for association with SCZ and BPAD. Sequence analysis of all exonic and flanking intronic regions of the SYNGR1 gene in 198 BPAD and 193 SCZ cases revealed a novel mutation Lsy99Glu (in one BPAD patient) and two other novel common polymorphisms [synonymous single nucleotide polymorphism (SNP--Ser97Ser) and an Asn ins/del] in the SYNGR1 gene. We also validated 9 out of 14 dbSNPs in our population. Case-control analysis revealed allelic (P = 0.028-0.00007) association of five polymorphisms with SCZ and/or BPAD cases. Further, 3-SNP (with LD block 1 SNPs) and 2-SNP (with LD block 2 SNPs) haplotype analyses did not show any association with either SCZ or BPAD. Our results support SYNGR1 as a probable susceptibility gene for SCZ and BPAD. Also, the observed association of SYNGR1 with both SCZ and BPAD suggests the likely involvement of a common pathway in the etiology of these disorders. PMID- 16215644 TI - Culturable bacterial symbionts isolated from two distinct sponge species (Pseudoceratina clavata and Rhabdastrella globostellata) from the Great Barrier Reef display similar phylogenetic diversity. AB - The diversity of the culturable microbial communities was examined in two sponge species-Pseudoceratina clavata and Rhabdastrella globostellata. Isolates were characterized by 16S rRNA gene sequencing and phylogenetic analysis. The bacterial community structures represented in both sponges were found to be similar at the phylum level by the same four phyla in this study and also at a finer scale at the species level in both Firmicutes and Alphaproteobacteria. The majority of the Alphaproteobacteria isolates were most closely related to isolates from other sponge species including alpha proteobacterium NW001 sp. and alpha proteobacterium MBIC3368. Members of the low %G + C gram-positive (phylum Firmicutes), high %G + C gram-positive (phylum Actinobacteria), and Cytophaga Flavobacterium-Bacteroides (phylum Bacteroidetes) phyla of domain Bacteria were also represented in both sponges. In terms of culturable organisms, taxonomic diversity of the microbial community in the two sponge species displays similar structure at phylum level. Within phyla, isolates often belonged to the same genus-level monophyletic group. Community structure and taxonomic composition in the two sponge species P. clavata and Rha. globostellata share significant features with those of other sponge species including those from widely separated geographical and climatic regions of the sea. PMID- 16215645 TI - Microcystins induce morphological and physiological changes in selected representative phytoplanktons. AB - Dissolved microcystins (MC) are regularly present in water dominated by microcystin-producing, bloom-forming cyanobacteria. In vitro experiments with environmentally feasible concentrations (5 x 10(-7) M) of the three most common microcystins, MC-LR, -RR, and -YR, revealed that they influence the metabolism of different representative phytoplanktons. At light intensities close to the cyanobacterial bloom environment (50 mumol m(-2) s(-1)), they produce morphological and physiological changes in both microcystin-producing and nonproducing Microcystis aeruginosa strains, and also have similar effects on the green alga Scenedesmus quadricauda that is frequently present in cyanobacterial blooms. All three microcystin variants tested induce cell aggregation, increase in cell volume, and overproduction of photosynthetic pigments. All three effects appear to be related to each other, but are not necessarily caused by the same mechanism. The biological activity of microcystins toward the light-harvesting complex of photobionts can be interpreted as a signal announcing the worsening of light conditions due to the massive proliferation of cyanobacteria. Although the function of microcystins is still unknown, it is evident that they have numerous effects on phytoplankton organisms in nature. These effects depend on the individual organism as well as on the various intracellular and extracellular signaling pathways. The fact that dissolved microcystins also influence the physiology of microcystin-producing cyanobacteria leads us to the conclusion that the role of microcystins in the producing cells differs from their role in the water environment. PMID- 16215646 TI - Bacterial colonization of the phyllosphere of mediterranean perennial species as influenced by leaf structural and chemical features. AB - In this study, we assessed various leaf structural and chemical features as possible predictors of the size of the phyllosphere bacterial population in the Mediterranean environment. We examined eight perennial species, naturally occurring and coexisting in the same area, in Halkidiki (northern Greece). They are Arbutus unedo, Quercus coccifera, Pistacia lentiscus, and Myrtus communis (evergreen sclerophyllous species), Lavandula stoechas and Cistus incanus (drought semi-deciduous species), and Calamintha nepeta and Melissa officinalis (non-woody perennial species). M. communis, L. stoechas, C. nepeta, and M. officinalis produce essential oil in substantial quantities. We sampled summer leaves from these species and (1) estimated the size of the bacterial population of their phyllosphere, (2) estimated the concentration of different leaf constituents, and (3) studied leaf morphological and anatomical features and expressed them in a quantitative way. The aromatic plants are on average more highly colonized than the other species, whereas the non-woody perennials are more highly colonized than the woody species. The population size of epiphytic bacteria is positively correlated with glandular and non-glandular trichome densities, and with water and phosphorus contents; it is negatively correlated with total phenolics content and the thickness of the leaf, of the mesophyll, and of the abaxial epidermis. No correlation was found with the density of stomata, the nitrogen, and the soluble sugar contents. By regression tree analysis, we found that the leaf-microbe system can be effectively described by three leaf attributes with leaf water content being the primary explanatory attribute. Leaves with water content >73% are the most highly colonized. For leaves with water content <73%, the phosphorus content, with a critical value of 1.34 mg g( 1) d.w., is the next explanatory leaf attribute, followed by the thickness of the adaxial epidermis. Leaves higher in phosphorus (>1.34 mg g(-1) d.w.) are more colonized, and leaves with the adaxial epidermis thicker than 20.77 microm are the least colonized. Although these critical attributes and values hold true only within the Mediterranean ecosystem studied and the range of observations taken, they are important because they provide a hypothesis to be tested in other Mediterranean ecosystems and other biomes. Such comparative studies may give insight as to the general properties governing the leaf-microbe system. PMID- 16215647 TI - Even conservation rules are made to be broken: implications for biodiversity. AB - Despite efforts to enclose and control conservation zones around the world, direct human impacts in conservation areas continue, often resulting from clandestine violations of conservation rules through outright poaching, strategic agricultural encroachment, or noncompliance. Nevertheless, next to nothing is actually known about the spatially and temporally explicit patterns of anthropogenic disturbance resulting from such noncompliance. This article reviews current understandings of ecological disturbance and conservation noncompliance, concluding that differing forms of noncompliance hold differing implications for diversity. The authors suggest that forms of anthropogenic patchy disturbance resulting from violation may maintain, if not enhance, floral diversity. They therefore argue for extended empirical investigation of such activities and call for conservation biologists to work with social scientists to assess this conservation reality by analyzing how and when incomplete enforcement and rule breaking drive ecological change. PMID- 16215648 TI - Public perception as a barrier to introducing wood in rivers for restoration purposes. AB - Reintroduction of wood in rivers for restoration purposes is now recognized in a positive way by scientists. Nevertheless, the perception of wood in riverscapes is strongly affected by the socio-cultural environment. This cultural influence might explain why wood reintroduction is accepted and promoted in some regions of the world but not in others, despite the demonstrated ecological benefits. From an extensive student perception survey, we show that most of the groups from nine countries in the world considered riverscapes with wood to be less aesthetic, more dangerous, and needing more improvement than riverscapes without wood. By contrast, this way of thinking was not observed in Germany, Sweden, and Oregon (USA), where the first instances of wood reintroduction occurred. PMID- 16215649 TI - Effects of short- and long-term disturbance resulting from military maneuvers on vegetation and soils in a mixed prairie area. AB - Loss of grassland species resulting from activities such as off-road vehicle use increases the need for models that predict effects of anthropogenic disturbance. The relationship of disturbance by military training to plant species richness and composition on two soils (Foard and Lawton) in a mixed prairie area was investigated. Track cover (cover of vehicle disturbance to the soil) and soil organic carbon were selected as measures of short- and long-term disturbance, respectively. Soil and vegetation data, collected in 1-m2 quadrats, were analyzed at three spatial scales (60, 10, and 1 m2). Plant species richness peaked at intermediate levels of soil organic carbon at the 10-m2 and 1-m2 spatial scales on both the Lawton and Foard soils, and at intermediate levels of track cover at all three spatial scales on the Foard soil. Species composition differed across the disturbance gradient on the Foard soil but not on the Lawton soil. Disturbance increased total plant species richness on the Foard soil. The authors conclude that disturbance up to intermediate levels can be used to maintain biodiversity by enriching the plant species pool. PMID- 16215650 TI - Purpose-driven public sector reform: the need for within-government capacity build for the management of slope stability in communities in the Caribbean. AB - This article stresses the importance of within-government capacity build as the optimal approach to minimizing landslide risk to the most vulnerable communities in the developing world. Landslide risk is an integrated issue that demands strong managerial leadership and multidisciplinary inclusion to develop structures that deliver sustainable improvements in the reduction of risk. The tension between projects demanding international technical and financial intervention and those capable of "within-country" solutions are examined. More particularly, the challenges of developing a management methodology capable of energizing inter-ministry collaboration to achieve community-level action is examined in the context of a recently established program of slope stability management in St. Lucia. The program, Management of Slope Stability in Communities (MoSSaiC), is shown to have successfully fostered not only extensive technical collaboration within government but also to have energized local communities in the shared mission of capacity build through their direct involvement in the management process. PMID- 16215651 TI - The use of an ecologic classification to improve water resource planning in New Zealand. AB - The Resource Management Act (RMA) legislates the management of most natural resources in New Zealand. The RMA invokes ecosystem-based management by requiring that regulation be based on managing the effects of resource according to "the life supporting capacity" of the environment. The management of water resources under the RMA is carried out at the regional level by regional councils. Regional councils can develop regional water plans to establish objectives and criteria for water management. Regional water planning under the RMA has been problematic, and regional plan objectives developed under the RMA have been criticized as too broad and not sufficiently quantified. As a consequence, many resource users are unconvinced of the need for the regulatory criteria promulgated by plans, whereas other groups are concerned that the environment is inadequately protected. This article proposes that a lack of ecologically relevant management units has prevented regional water plans from fulfilling their intended function under the RMA. Then it introduces the use of River Environment Classification as a means of defining units for assessment and management, and provides three case studies that demonstrate its potential to support regional water management planning. The discussion shows that the specificity of regional assessments can be increased if ecologic variation is stratified into distinctive units (i.e., units within which variation in the characteristics of interest is reduced) as part of the assessment process. The increased specificity of the assessments increases the possibility that regional objectives and criteria for water management can be derived that are quantitative and justifiable and that provide certainty for stakeholders. The authors conclude that greater choice and meaning can be generated in regional planning processes if regional variation in ecologic characteristics is stratified using a classification, and if classes are used as units for assessment and management. PMID- 16215652 TI - Urbanization and the loss of resource lands in the chesapeake bay watershed. AB - We made use of land cover maps, and land use change associated with urbanization, to provide estimates of the loss of natural resource lands (forest, agriculture, and wetland areas) across the 168,000 km2 Chesapeake Bay watershed. We conducted extensive accuracy assessments of the satellite-derived maps, most of which were produced by us using widely available multitemporal Landsat imagery. The change in urbanization was derived from impervious surface area maps (the built environment) for 1990 and 2000, from which we estimated the loss of resource lands that occurred during this decade. Within the watershed, we observed a 61% increase in developed land (from 5,177 to 8,363 km2). Most of this new development (64%) occurred on agricultural and grasslands, whereas 33% occurred on forested land. Some smaller municipalities lost as much as 17% of their forest lands and 36% of their agricultural lands to development, although in the outlying counties losses ranged from 0% to 1.4% for forests and 0% to 2.6% for agriculture. Fast-growing urban areas surrounded by forested land experienced the most loss of forest to impervious surfaces. These estimates could be used for the monitoring of the impacts of development across the Chesapeake Bay watershed, and the approach has utility for other regions nationwide. In turn, the results and the approach can help jurisdictions set goals for resource land protection and acquisition that are consistent with regional restoration goals. PMID- 16215653 TI - Use of animal species data in environmental impact assessments. AB - Environmental Impact Assessments (EIAs) should ideally help minimize adverse effects on biological diversity by considering impacts of projects on wide ranges of species. This paper investigates how recent Dutch EIAs included the species comprising animal diversity. We present results of two studies on fauna data used in the EIAs. Objectives were to determine for different taxa (a) the relative representation of species in Environmental Impact Statements (EISs); (b) the extent to which EISs referred to specific species and the accuracy of survey data referred to; and (c) apparent roles of different EIA actors in species inclusion. EIAs were found to use data on various taxa but on limited numbers of species. The frequency with which taxa were included varied significantly. Birds were most frequently included, followed by mammals, amphibians, and other species groups. The quality of data on birds exceeded that regarding other vertebrates. Our results indicate that (a) EIA working groups of independent experts were the most influential in determining the data to be used; (b) on average, proponents included data more often than required by guidelines; and (c) in 30 to 40% of the EIAs, the participation of nongovernmental organizations prompted use of data. Despite the key role of experts in data inclusion, the taxon rankings found in the EIAs showed little deviation from those observed in studies on people's preferences for species. Given the limited ranges of species considered, it is doubtful that the EIAs examined effectively contributed to conserving animal species diversity. PMID- 16215654 TI - Phytoremediation of soil polluted by nickel using agricultural crops. AB - Soil pollution due to heavy metals is widespread; on the world scale, it involves about 235 million hectares. The objectives of this research were to establish the uptake efficiency of nickel by some agricultural crops. In addition, we wanted to establish also in which part of plants the metal is stored for an eventual use of biomass or for recycling the metal. The experiments included seven herbaceous crops such as: barley (Hordeum vulgaris), cabbage (Brassica juncea), spinach (Spinacea oleracea), sorghum (Sorgum vulgare), bean (Phaseolus vulgaris), tomato (Solanum lycopersicum), and ricinus (Ricinus communis). We used three levels of treatment (150, 300, and 600 ppm) and one control. At the end of the biological cycle of the crops, the different parts of plants, i.e., roots, stems, leaves, fruits, or seeds, were separately collected, oven dried, weighed, milled, and separately analysed. The leaves and stems of spinach showed a very good nickel storage capacity. The ricinus too proved to be a very good nickel storer. The ability of spinach and ricinus to store nickel was observed also in the leaves of cabbage, even if with a lower storage capacity. The bean, barley, and tomato, in decreasing order of uptake and storage capacity, showed a high concentration of nickel in leaves and stems, whereas the sorghum evidenced a lesser capacity to uptake and store nickel in leaves and stems. The bean was the most efficient in storing nickel in fruits or grains. Tomato, sorghum, and barley have shown a storage capacity notably less than bean. The bean appeared to be the most efficient in accumulating nickel in the roots, followed in decreasing order by sorghum, ricinus, and tomato. With regard to the removal of nickel, spinach was the most efficient as it contains the highest level of this metal per gram of dry matter. The ricinus, cabbage, bean, sorghum, barley, and tomato evidenced a progressively decreasing efficiency in the removal of nickel. PMID- 16215655 TI - Use of microbial community to evaluate performance of a wetland system in treating Pb/Zn mine drainage. AB - The performance of a wetland system in treating lead (Pb)/zinc (Zn) mine drainage was evaluated by using the polyurethane foam unit (PFU) microbial community (method), which has been adopted by China as a standardized procedure for monitoring water quality. The wetland system consisted of four cells with three dominant plants: Typha latifolia, Phragmites australis and Paspalum distichum. Physicochemical characteristics [pH, EC, content of total suspended solid (TSS) and metals (Pb, Zn, Cd, and Cu)] and PFU microbial community in water samples had been investigated from seven sampling sites. The results indicated that the concentrations of Pb, Zn, Cd, Cu, and TSS in the mine drainage were gradually reduced from the inlet to the outlet of the wetland system and 99%, 98%, 75%, 83%, and 68% of these metals and TSS respectively, had been reduced in concentration after the drainage passed through the wetland system. A total of 105 protozoan species were identified, the number of protozoa species and the diversity index (DI) gradually increased, while the heterotrophic index (HI) gradually decreased from the inlet to the outlet of the wetland system. The results indicated that DI, HI, and total number species of protozoa could be used as biological indicators indicating the improvement of water quality. PMID- 16215656 TI - Validation study of a nonspecific psychological distress scale. AB - BACKGROUND: Psychological distress scales are often used in national epidemiological surveys to monitor the mental health status and predict demands in mental health services. These scales have the advantage of being easy to administer and inexpensive to use. The goal of this study is to assess the clinical validity of the Psychological Distress Manifestations Measure Scale (PDMMS) by comparing it to a standard criterion. METHOD: The validation study is based on data from a large-scale mental health survey conducted in 1999 in the Montreal area (Canada). The target population was constituted of adults living in private households. A telephone survey was carried out with a probability sample of 4,704 respondents using the Composite International Diagnostic Interview Simplified (CIDIS) to detect mental disorders. Then, subsequent face-to-face interviews with a subsample of 359 of these respondents were conducted to validate other measures for assessing mental health needs for care and services including the PDMMS. RESULTS: Our study showed that high psychological distress is highly associated with mental disorder (OR=5.94). However, a large majority of the people in the high psychological distress category does not have a known mental problem. CONCLUSIONS: These data confirm that like other psychological distress scales, the PDMMS is not a diagnostic tool. Rather, it is designed to explore comorbidity among symptoms, independent of caseness. The prevalence of psychological distress in the population allows us to identify people who have subclinical symptoms substantial enough to precipitate dysfunctioning in everyday life and who utilize health services more frequently. The use of this tool for epidemiological surveys is useful for mental health service planning because it provides information on the needs of individuals whose state of mental health affects social functioning even though they do not suffer from pathology. PMID- 16215658 TI - Evaluation of "depression in primary care" innovations. AB - Key stakeholders and executive decision makers in health care system require clear and convincing data of the value of chronic illness care models for the primary care treatment of depression. Well-conceived and conducted evaluations provide this necessary information. This case study describes the experience of a large, nonprofit health care system's experience with implementing and evaluating a quality improvement program for integrating depression management into primary care. The commentary that follows discusses specific evaluation questions that are relevant to each of the stakeholder groups involved in deciding whether or not to continue supporting such programs. PMID- 16215657 TI - Changes in the edentate mandible in the elderly. AB - Resorption of alveolar bone is the best recognized feature of mandibular aging in the edentate subject. The other consequences of the loss of teeth in the elderly are less well known. An anthropometric study of the mandible by antero-posterior and lateral radiographs of subjects older than 70 years both dentate and edentate but without any maxillo-mandibular dysmorphosis has been done to demonstrate the differences, which exist between the dentate and edentate mandible. The edentate mandibles showed a diminution in the height of the symphysis and increase in the height of the mandibular incisure. A diminution in the height of the body and an increase in the gonial angle in the significant manner. No significant difference was seen for the height of the ramus and the length of the mandible, the minimum width of the ramus and the bigonial width. The diminution in the height of the mandibular symphysis and of the body is explained by the resorption of the alveoli part of the mandible. The increase in the mandibular angle and the diminution in the height of the mandibular incisure may be explained by disequilibrium between the elevator and depressor muscles of the mandible, as a function of the elevator muscles or by the absence of the molar buttress. PMID- 16215659 TI - Patient self-management in the primary care treatment of depression. AB - Efforts to improve primary care treatment of depression incorporate elements of the chronic illness care model, including patient self-management strategies. Case studies, focus groups and the literature suggest six key components of depression self-management programs: (1) implement behavioral change interventions, (2) plan for crisis and relapse prevention, (3) re-establish personal meaning, (4) attend to patients' experience, context and community, (5) build a patient-clinician partnership and (6) create an integrated, self management support structure. Successful implementation of these components is facilitated by (1) the care system's collective and empathic understanding of the disease itself; (2) sufficient time; (3) adequate funding and (4) robust clinical information systems. PMID- 16215660 TI - Challenges of implementing depression care management in the primary care setting. AB - Empirical evidence shows that care management is an effective tool for improving depression treatment in primary care patients. However, several conceptual and practical issues have not been sufficiently addressed. This article explores questions concerning the scope of care management services within the chronic illness care model; optimal ways to identify depressed patients in the primary care setting; responsibilities and desirable qualifications of depression care managers; the location and manner in which care managers interact with patients; costs of services provided by care managers; and the level of supervision by mental health specialists that is necessary to ensure quality care. PMID- 16215661 TI - The role of clinical information technology in depression care management. AB - We examine the literature on the growing application of clinical information technology in managing depression care and highlight lessons learned from Robert Wood Johnson Foundation's national program "Depression in Primary Care-Incentives Demonstrations." Several program sites are implementing depression care registries. Key issues discussed about implementing registries include using a simple yet functional format, designing registries to track multiple conditions versus depression alone (i.e., patient-centric versus disease-centric registries) and avoiding violations of patient privacy with the advent of more advanced information technologies (e.g., web-based formats). Finally, we discuss some implications of clinical information technology for health care practices and policy makers. PMID- 16215662 TI - Implementation of guideline-based care for depression in primary care. AB - Evidence-based clinical practice guidelines for treating depression in primary care settings were developed, in part, to ensure that health services are provided in a consistent, high-quality, and cost-effective manner. Yet for a variety of reasons, guideline-based primary care for depression remains the exception rather than the rule. This work provides a brief review of effective strategies used to customize and then deliver evidence-based treatment for depression in primary care settings; describes two representative case studies that illustrate locally customized collaborative care strategies for treatment delivery; and concludes with principles and implications for policy and practice based on our practical experiences. PMID- 16215663 TI - Commentary: an economic perspective on implementing evidence-based depression care. AB - Despite the development of cost-effective evidence-based models for treating depression in primary care, economic and organizational barriers often impede sustainability in routine clinical practice. Under the Depression in Primary Care: Linking Clinical and System Strategies program, the Robert Wood Johnson Foundation (RWJF) funded eight demonstration grants to implement clinical changes in depression care alongside changes in contractual relationships, payment methods and other economic arrangements. The preceding articles summarize the specific economic and organizational changes implemented under four of these demonstration projects. This commentary highlights certain elements that appear critical to successfully re-align system incentives to support evidence-based depression care based in part on the experiences of these four sites. PMID- 16215664 TI - Realigning economic incentives for depression care at UCSF. AB - Behavioral health carve-out arrangements create financial disincentives for primary care providers (PCPs) to treat depression. A novel collaboration between a primary care practice, a health insurer, and a managed behavioral health organization (MBHO) allows PCPs to receive reimbursement and schedule longer appointments to care for depressed patients. This article describes the details of the arrangement, and early results of this collaboration. Early results find that financial incentives are critical for implementation, but that time incentives do not appear to motivate PCPs. Sustainability of this model will require participation of multiple primary care practices, health insurers, and MBHOs. PMID- 16215665 TI - The economics of depression in primary care: de-fragmentation in the Oregon Medicaid market. AB - The Oregon Medicaid program legislatively separates the administration of physical health and mental health services, even though behavioral and physical health conditions significantly impact each other. To overcome this barrier and enhance integrated care, CareOregon, a large Medicaid only health plan partnered with two of its largest provider groups to pilot two different models of integration. In one, an "ownership" model, behavioral health specialists were employed by Federally Qualified Health Center primary care clinics and functioned in a common care model with other providers. In the other, a "loaned" model, behavioral specialists were placed in primary care clinics by community mental health centers and continued to function in a specialty mental health model. The qualitative effects of these two models are discussed. PMID- 16215666 TI - Realigning clinical and economic incentives to support depression management within a medicaid population: the Colorado access experience. AB - The authors describe their experiences in developing an economically sustainable depression care management program within Colorado Access, a non-profit Medicaid health plan. They describe high rates of mental health issues, medical comorbidities, and psychosocial barriers to care within the plan's Medicaid population. They discuss how the company redirected resources to incorporate depression care management into an intensive care management program focused on high-cost members with multiple chronic medical conditions. This strategy allowed Colorado Access to cost effectively care manage a targeted group of high-cost Medicaid recipients across multiple primary care physician (PCP) practices without requiring changes in provider workflow. PMID- 16215668 TI - Depression in primary care: learning lessons in a national quality improvement program. PMID- 16215667 TI - Depression in primary care: learning lessons in a national quality improvement program. AB - In this introduction, we describe the overall context and rationale for the Depression in Primary Care program and the design and implementation of its key components, especially emphasizing its unique combined clinical and economic/systems framework. We also discuss some of the new challenges and opportunities that may impact the program's evolution and the state of behavioral health care more generally. We conclude with some thoughts on potential future scenarios and strategies for improving the quality of behavioral health care, including the treatment of depression in primary care. PMID- 16215669 TI - Assessment of the downstream portion of the mitochondrial pathway of caspase activation in patients with acute myeloid leukemia. AB - Most chemotherapeutic agents used in the treatment of acute myeloid leukemia (AML) induce apoptosis by triggering the mitochondrial pathway of caspase activation. To investigate the downstream portion of the mitochondrial pathway of caspase activation in patients with AML, cytosolic lysates were stimulated with cytochrome c and dATP and hydrolysis of Ac-DEVD-AFC by effector caspases was measured. Defects in the distal mitochondrial pathway were more common in samples from patients with AML that relapsed rapidly after induction chemotherapy compared to samples from treatment naive patients. The incidence of blocked pathways did not differ based on response to induction chemotherapy, as even nonresponders generally had an intact pathway. When the distal mitochondrial pathway was blocked, defects were usually at the level of the effector caspases. Thus, functional defects in the distal portion of the mitochondrial pathway of caspase activation may help explain the nature of response and relapse after treatment. PMID- 16215670 TI - Bcl-2 attenuates anticancer agents-induced apoptosis by sustained activation of Akt/protein kinase B in U937 cells. AB - Aberrant overexpression of antiapoptotic members of the Bcl-2 protein family contributes to resistance to anticancer therapeutic drugs. Thus, this protein represent attractive target for novel anticancer agents. In the present study, we determined the effect of the anti-apoptosis protein Bcl-2 on caspase-3 activation, PLC-gamma1 degradation and Akt activation during the various anticancer agents-induced apoptosis. Treatment with chrysin for 12 h produced morphological features of apoptosis in U937 cells, which was associated with caspase-3 activation and PLC-gamma1 degradation. Induction of apoptosis was also accompanied by down-regulation of XIAP and inactivation of Akt. Chrysin-induced caspase-3 activation, PLC-gamma1 degradation and apoptosis were significantly attenuated in Bcl-2 overexpressing U937/Bcl-2 cells. Ectopic expression of Bcl-2 appeared to inhibit ceramide-, and Akt specific inhibitor (SH-6)-induced apoptosis by sustained Akt activation. Thus, our findings imply that some of the biological functions of Bcl-2 may be attributed to their ability to inhibit anticancer agents-induced apoptosis through the sustained Akt activation. PMID- 16215671 TI - Biphasic behavior of changes in elemental composition during staurosporine induced apoptosis. AB - Although the identification of events that occur during apoptosis is a fundamental goal of apoptotic cell death research, little is know about the precise sequence of changes in total elemental composition during apoptosis. We evaluated total elemental composition (Na, Mg, P, Cl, S, and K) in relation to molecular and morphological features in human U937 cells induced to undergo apoptosis with staurosporine, an intrinsic pathway activator. To evaluate total elemental content we used electron probe X-ray microanalysis to measure simultaneously all elements from single, individual cells. We observed two phases in the changes in elemental composition (mainly Na, Cl and K). The early phase was characterized by a decrease in intracellular K (P<0.001) and Cl (P<0.001) content concomitant with cell shrinkage, and preceded the increase in proteolytic activity associated with the activation of caspase-3. The later phase started with caspase-3 activation, and was characterized by a decrease in the K/Na ratio (P<0.001) as a consequence of a significant decrease in K and increase in Na content. The inversion of intracellular K and Na content was related with the inhibition of Na+/K+ ATPase. This later phase was also characterized by a significant increase (P<0.001) in intracellular Cl with respect to the early phase. In addition, we found a decrease in S content and an increase in the P/S ratio. These distinctive changes coincided with chromatin condensation and DNA fragmentation. Together, these findings support the concept that changes in total elemental composition take place in two phases related with molecular and morphological features during staurosporine-induced apoptosis. PMID- 16215672 TI - Modulation of Fas-induced apoptosis by p75 neurotrophin receptor in a human neuroblastoma cell line. AB - Fas and p75 neurotrophin receptors (p75(NTR)) are death receptors that alone induce apoptosis of SH-SY5Y neuroblastoma cell line respectively by Fas ligand or brain-derived neurotrophic factor (BDNF, a p75(NTR) ligand). We report on the modulation of Fas-mediated apoptosis by concomitant p75(NTR) activation. The exposure to both ligands suppressed the apoptotic effect. A co-localisation of Fas and p75(NTR) receptors was evidenced by co-capping and immunoprecipitation assays. Moreover, a caspase-8 inhibitor suppressed the protective effect of the concomitant BDNF and Fas ligand stimulation, suggesting that p75(NTR) and Fas receptors could share common signalling pathways. PMID- 16215673 TI - Sensitization of prostate carcinoma cells to Apo2L/TRAIL by a Bcl-2 family protein inhibitor. AB - Overexpression of anti-apoptotic Bcl-2 family proteins may play an important role in the aggressive behavior of prostate cancer cells and their resistance to therapy. The Bcl-2 homology 3 domain (BH3) is a uniquely important functional element within the pro-apoptotic class of the Bcl-2-related proteins, mediating their ability to dimerize with other Bcl-2-related proteins and promote apoptosis. The BH3 inhibitors (BH3Is) function by disrupting the interactions mediated by the BH3 domain between pro- and anti-apoptotic members of the Bcl-2 family and liberating more Bax/Bak to induce mitochondrial membrane permeabilization. LNCaP-derived C4-2 human prostate cancer cells are quite resistant to non-tagged, human recombinant soluble Apo2 ligand [Apo2L, also Tumor necrosis factor (TNF)-related apoptosis-inducing ligand, TRAIL], a tumor specific drug that is now in clinical trials. However, when Apo2L/TRAIL was combined with the Bcl-xL inhibitor, BH3I-2', it induced apoptosis synergistically through activation of Caspase-8 and the proapoptotic Bcl-2 family member Bid, resulting in the activation of effector Caspase-3 and proteolytic cleavage of Poly(ADP ribose) polymerase, events that were blocked by the pan-caspase inhibitor zVAD fmk. Our data indicate that, in combination with the BH3 mimetic, BH3I-2', Apo2L/TRAIL synergistically induces apoptosis in C4-2 human prostate cancer cells through both the extrinsic and intrinsic apoptotic pathways. PMID- 16215674 TI - Cathepsin B inactivation attenuates the apoptotic injury induced by ischemia/reperfusion of mouse liver. AB - BACKGROUND: A major mechanism underlying warm ischemia/reperfusion (I/R) injury during liver transplantation is the activation of the caspase chain, which leads to apoptosis. Recently, it was demonstrated that the release of cathepsin B, a cysteine protease, from the cytosol in liver injury induces mitochondrial release of cytochrome c and the activation of caspase-3 and -9, thereby leading to apoptosis. The aim of this study was to ascertain if cathepsin B inactivation attenuates the apoptotic injury due to I/R in mouse liver. METHODS: A model of segmental (70%) hepatic ischemia was used. Eighteen mice were anesthetized and randomly divided into three groups: (1) CONTROL GROUP: sham operation (laparotomy); (2) Ischemic group: midline laparotomy followed by occlusion of all structures in the portal triad to the left and median lobes for 60 min (ischemic period); (3) STUDY GROUP: like group 2, but with intraperitoneal administration of a pharmacological inhibitor of cathepsin B (4 mg/100 g) 30 min before induction of ischemia. Serum liver enzyme levels were measured by biochemical analysis, and intrahepatic caspase-3 activity was measured by fluorometric assay; apoptotic cells were identified by morphological criteria, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorometric assay, and immunohistochemistry for caspase-3. RESULTS: Showed that at 6 h of reperfusion, there was a statistically significant reduction in liver enzyme levels in the animals pretreated with cathepsin B inhibitor (p<0.05). On fluorometric assay, caspase-3 activity was significantly decreased in group 3 compared to group 2 (p<0.0001). The reduction in postischemic apoptotic hepatic injury in the cathepsin B inhibitor -treated group was confirmed morphologically, by the significantly fewer apoptotic hepatocyte cells detected (p<0.05); immunohistochemically, by the significantly weaker activation of caspase-3 compared to the ischemic group (p<0.05); and by the TUNEL assay (p<0.05). CONCLUSION: The administration of cathepsin B inhibitor before induction of ischemia can attenuate postischemic hepatocyte apoptosis and thereby minimize liver damage. Apoptotic hepatic injury seems to be mediated through caspase-3 activity. These findings have important implications for the potential use of cathepsin B inhibitors in I/R injury during liver transplantation. PMID- 16215675 TI - Chemopreventive agent-induced modulation of death receptors. AB - The goals of chemoprevention of cancer are to inhibit the initiation or suppress the promotion and progression of preneoplastic lesions to invasive cancer through the use specific natural or synthetic agents. Therefore, a more desirable and aggressive approach is to eliminate aberrant clones by inducing apoptosis rather than merely slowing down their proliferation. The increased understanding of apoptosis pathways has directed attention to components of these pathways as potential targets not only for chemotherapeutic but also for chemopreventive agents. Activation of death receptors triggers an extrinsic apoptotic pathway, which plays a critical role in tumor immunosurveillance. An increasing number of previously identified chemopreventive agents were found to induce apoptosis in a variety of premalignant and malignant cell types in vitro and in a few animal models in vivo. Some chemopreventive agents such as non-steroidal anti inflammatory drugs, tritepenoids, and retinoids increase the expression of death receptors. Thus, understanding the modulation of death receptors by chemopreventive agents and their implications in chemoprevention may provide a rational approach for using such agents alone or in combination with other agents to enhance death receptor-mediated apoptosis as a strategy for effective chemoprevention of cancer. PMID- 16215676 TI - Photodynamic therapy-induced death of HCT 116 cells: Apoptosis with or without Bax expression. AB - Cell death following photodynamic therapy (PDT) with the photosensitizer Pc 4 involves the intrinsic pathway of apoptosis. To evaluate the importance of Bax in apoptosis after PDT, we compared the PDT responses of Bax-proficient (Bax(+/-)) and Bax knock-out (BaxKO) HCT116 human colon cancer cells. PDT induced a slow apoptotic process in HCT Bax(+/-) cells following a long delay in the activation of Bax and release of cytochrome c from mitochondria. Although cytochrome c was not released from mitochondria following PDT in BaxKO cells, an alternative mechanism of caspase-dependent apoptosis with extensive chromatin and DNA degradation was found in these cells. This alternative process was less efficient and slower than the normal apoptotic process observed in Bax(+/-) cells. Early events upon PDT, such as the loss of mitochondrial membrane potential, photodamage to Bcl-2, and activation of p38 MAP kinase, were observed in both HCT116 cell lines. In spite of differences in the efficiency and mode of apoptosis induced by PDT in the Bax(+/-) and BaxKO cells, they were found to be equally sensitive to killing by PDT, as determined by loss of clonogenicity. Thus, for Pc 4-PDT, the commitment to cell death occurs prior to and independent of Bax activation, but the process of cellular disassembly differs in Bax expressing vs. non-expressing cells. PMID- 16215677 TI - Inhibition of TRAIL-induced apoptosis by IL-8 is mediated by the p38-MAPK pathway in OVCAR3 cells. AB - INTRODUCTION: TRAIL (TNF-Related Apoptosis Inducing Ligand) is a member of the TNF superfamily of cell death inducing ligands. Interestingly, while malignant cells are responsive to TRAIL-induced cell death when used alone or in combination with other agents, normal cells do not appear to be sensitive to this ligand, making it a desirable therapeutic compound against many cancers, including many ovarian carcinomas. Interleukin-8 (IL-8), a member of the C-X-C chemokine family, has been found to be at significantly higher level in the ascites from patients with ovarian cancer. We have previously demonstrated a role for IL-8 in blocking TRAIL's ability to induce apoptosis in the ovarian cancer cell line, OVCAR3, possibly by repressing the DR4 TRAIL receptor expression and blocking caspase-8 cleavage. In addition, we showed a member of the mitogen activated protein kinase (MAPK) superfamily, p38gamma, is among the genes regulated in OVCAR3 cells by TRAIL and IL-8. The present study further investigates involvement of the p38 MAPK pathway in IL-8's ability to block TRAIL induced apoptosis in the ovarian surface epithelial cancer cell line, OVCAR3. RESULTS: In this study we demonstrate that p38gamma as well as p38alpha play a significant role in IL-8's ability to block TRAIL-induced apoptosis. Through array analysis, as well as confirmation with other methods, we detected regulation of p38gamma and p38alpha following treatment of the cancer cell line with IL-8 or TRAIL. We also tested two other isoforms of p38 MAPK, p38beta and p38delta, but did not find significant regulation by IL-8 or TRAIL. We also examined activation of the p38 MAPK pathway, up-stream as well as down-stream, and noticed activation of the pathway following treatment with TRAIL and decreased activity when IL-8 was introduced. With the use of specific inhibitors, we were able to further confirm the role of this pathway in TRAIL-induced apoptosis, and IL-8's ability to block this apoptosis, in ovarian cancer cell lines. CONCLUSION: Taken together, these results further solidify the role of IL 8 in blocking the TRAIL-induced apoptosis in these ovarian carcinoma cells and provide new molecular insight into this potentially important therapeutic target. PMID- 16215679 TI - Role of mitochondria in tamoxifen-induced rapid death of MCF-7 breast cancer cells. AB - Tamoxifen (Tam) is widely used in chemotherapy of estrogen receptor-positive breast cancer. It inhibits proliferation and induces apoptosis of breast cancer cells by estrogen receptor-dependent modulation of gene expression, but recent reports have shown that Tam (especially at pharmacological concentrations) has also rapid nongenomic effects. Here we studied the mechanisms by which Tam exerts rapid effects on breast cancer cell viability. In serum-free medium 5-7 microM Tam induced death of MCF-7 and MDA-MB-231 cells in a time-dependent manner in less than 60 min. This was associated with release of mitochondrial cytochrome c, a decrease of mitochondrial membrane potential and an increase in production of reactive oxygen species (ROS). This suggests that disruption of mitochondrial function has a primary role in the acute death response of the cells. Accordingly, bongkrekic acid, an inhibitor of mitochondrial permeability transition, was able to protect MCF-7 cells against Tam. Rapid cell death induction by Tam was not associated with immediate activation of caspase-9 or cleavage of poly (ADP-ribose) polymerase. It was not blocked by the caspase inhibitor z-Val-Ala-Asp-fluoromethylketone either. Diphenylene ionodium (DPI), an inhibitor of NADPH oxidase, was able to prevent Tam-induced cell death but not cytochrome c release, which suggests that ROS act distal to cytochrome c. The pure antiestrogen ICI 182780 (1 microM) could partly oppose the effect of Tam in estrogen receptor positive MCF-7 cells, but not in estrogen receptor negative MDA MB-231 cells. Pre-culturing MCF-7 cells in the absence of 17beta-estradiol (E(2)) or in the presence of a low Tam concentration (1 microM) made the cells even more susceptible to rapid death induction by 5 or 7 microM Tam. This effect was associated with decreased levels of the anti-apoptotic proteins Bcl-X(L) and Bcl 2. In conclusion, our results demonstrate induction of a rapid mitochondrial cell death program in breast cancer cells at pharmacological concentrations of Tam, which are achievable in tumor tissue of Tam-treated breast cancer patients. These mechanisms may contribute to the ability of Tam therapy to induce death of breast cancer cells. PMID- 16215678 TI - Intestinal intraepithelial lymphocyte derived angiotensin converting enzyme modulates epithelial cell apoptosis. AB - BACKGROUND & AIMS: Intestinal adaptation in short bowel syndrome (SBS) consists of increased epithelial cell (EC) proliferation as well as apoptosis. Previous microarray analyses of intraepithelial lymphocytes (IEL) gene expression after SBS showed an increased expression of angiotensin converting enzyme (ACE). Because ACE has been shown to promote alveolar EC apoptosis, we examined if IEL derived ACE plays a role in intestinal EC apoptosis. METHODS: Mice underwent either a 70% mid-intestinal resection (SBS group) or a transection (Sham group) and were studied at 7 days. ACE expression was measured, and ACE inhibition (ACE I, enalaprilat) was used to assess ACE function. RESULTS: IEL-derived ACE was significantly elevated in SBS mice. The addition of an ACE-I to SBS mice resulted in a significant decline in EC apoptosis. To address a possible mechanism, tumor necrosis factor alpha (TNF-alpha) mRNA expression was measured. TNF-alpha was significantly increased in SBS mice, and decreased with ACE-I. Interestingly, ACE I was not able to decrease EC apoptosis in TNF-alpha knockout mice. CONCLUSIONS: This study shows a previously undescribed expression of ACE by IEL. SBS was associated with an increase in IEL-derived ACE. ACE appears to be associated with an up-regulation of intestinal EC apoptosis. ACE-I significantly decreased EC apoptosis. PMID- 16215680 TI - Reactive oxygen species induce signals that lead to apoptotic DNA degradation in primary CD4+ T cells. AB - Reactive oxygen species are toxic to cells but they may also have active roles in transducing apoptotic events. To study the role of reactive oxygen species in growth factor depletion induced apoptosis of human primary CD4+ T cells, we used a synthetic manganese porphyrin superoxide dismutase mimetic to detoxify superoxide anions formed during apoptosis. Apoptosis of primary CD4+ T cells was characterized by generation of superoxide anions, plasma membrane phosphatidyl serine translocation, loss of mitochondrial membrane potential, activation of caspase 3, condensation of chromatin, as well as DNA degradation. The detoxification of superoxide anions did not influence plasma membrane phosphatidyl-serine translocation, or chromatin condensation, and only marginally inhibited the loss of mitochondrial membrane potential and the formation of DNA strand breaks. In contrast, the detoxification of superoxide anions significantly reduced caspase 3 activity and almost completely inhibited the apoptotic decrease in total cellular DNA content as measured by propidium iodide staining. Our results indicate that reactive oxygen anions induce signals leading to efficient DNA degradation after the initial formation of DNA strand breaks. Thus, reactive oxygen anions have active roles in signaling that lead to the apoptotic events. PMID- 16215681 TI - Prostaglandins and activation of AC/cAMP prevents anoikis in IEC-18. AB - Recent data indicates that chronic inflammation of the intestine such as Crohn's or ulcerative colitis puts those individuals at heightened risk for colorectal adenocarcinoma. In this study, we examine the effect of the inflammatory mediator PGE(2) and associated signalling on detachment-induced cell death (anoikis) in intestinal epithelial cells. Treatment of detached IEC-18 with 0.01-0.05 microM PGE(2) increased cell viability as well as induced aggregation. As EP4 prostaglandin receptors on IEC are coupled to adenylate cyclase, we next treated cells with agents that promote cAMP signalling (Forskolin, dbcAMP, and etazolate), all of which promoted IEC aggregation as well as survival. We next treated detached IECs with specific inhibitors of adenylate cyclase or PKA, which accelerated anoikis. To explore the mechanism of cell-cell adhesion, we next treated detached IECs with an anti-E-cadherin blocking antibody which dispersed aggregates induced by dbcAMP, and an adenovirus expressing a dominant negative E cadherin (EcadDeltaEC) prevented aggregate formation. Interestingly EcadDeltaEC prevented aggregation of IEC induced by dbcAMP but did not significantly reduce viability. This suggests that cAMP signalling is important in both aggregate formation and promoting viability but these are distinct events. Taken together, these data support a mechanism whereby elevated PGE(2) levels characteristic of colitis prevent anoikis by activating an AC-, cAMP-, and PKA-dependent signalling pathway. The delay of apoptosis by PGE(2) may be one mechanism by which inflammation may contribute to carcinogenesis. PMID- 16215682 TI - Cell death induced by ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid in anaplastic thyroid carcinoma cells is via a mitochondrial-mediated pathway. AB - The chemical compound ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F), isolated from the Chinese herbal medicine plant Pteris semipinnata L, has been known to exert antitumor activity. However, the molecular mechanism of the action is not understood. In this study we demonstrated that apoptotic cell death induced by 5F in FRO cells was concentration- and time-dependent. The rapid increase in intracellular reactive oxygen species (ROS) levels was involved in the mechanism of cell death. c-Jun N-terminal kinase (JNK) activation and G2 block were related to cell death induced by 5F. Extracellular signal-related kinase (ERK) and p38 were also activated, but as survival signals in response to 5F treatment to counteract the induction of cell death. In the process of the induction of apoptotic cell death, Bax translocated into mitochondria, a reduction in Delta psi(m) was observed and a release of cytochrome c and apoptosis inducing factor (AIF) from mitochondria into the cytosol occurred, indicating that cell death induced by 5F was through a mitochondrial-mediated pathway. PMID- 16215683 TI - Upstream control of apoptosis by caspase-2 in serum-deprived primary neurons. AB - During development as well as in pathological situations, neurons that fail to find appropriate targets or neurotrophic factors undergo cell death. Using primary cortical neurons subjected to acute serum-deprivation (SD), we have examined caspases activation, mitochondrial dysfunction and cell death parameters. Among a panel of metabolic, signaling and caspases inhibitors only those able to interfere with caspase-2 like activity protect primary neurons against SD-induced cell death. In situ detection and subcellular fractionation demonstrate a very early activation of cytosolic caspase-2, which controls Bax cleavage, relocalization and mitochondrial membrane permeabilization (MMP). Both z-VDVAD-fmk and a siRNA specific for caspase-2 abolish Bax changes, mitochondrial membranes permeabilization, as well as cytochrome c release-dependent activation of caspase-9/caspase-3, nuclear alterations, phosphatidylserine exposure, neurites dismantling and neuronal death. Hence, caspase-2 is an early checkpoint for apoptosis initiation in primary neurons subjected to serum deprivation. PMID- 16215684 TI - Mechanisms of induction of apoptosis by anthraquinone anticancer drugs aclarubicin and mitoxantrone in comparison with doxorubicin: relation to drug cytotoxicity and caspase-3 activation. AB - We examined molecular events and morphological features associated with apoptosis induced by anthraquinone anticancer drugs aclarubicin, mitoxantrone and doxorubicin in two spontaneously immortalized cell lines (NIH 3T3 and B14) in relation to cytotoxicity of these drugs. The investigated cells showed similar sensitivity to aclarubicin but different sensitivity to doxorubicin and mitoxantrone: mitoxantrone was the most cytotoxic drug in both cell lines. All three drugs triggered both apoptosis and necrosis but none of these processes was positively correlated with their cytotoxicity. Apoptosis was the prevalent form of cell kill by aclarubicin, while doxorubicin and mitoxantrone induced mainly the necrotic mode of cell death. The extent and the timing of apoptosis were strongly dependent on the cell line, the type of the drug and its dose, and were mediated by caspase-3 activation. A significant increase in caspase-3 activity and the percentage of apoptotic cells, oligonucleosomal DNA fragmentation, chromatin condensation and formation of apoptotic bodies was observed predominantly in B14 cells. NIH 3T3 cells showed lesser changes and a lack of DNA fragmentation. Aclarubicin was the fastest acting drug, inducing DNA fragmentation 12 h earlier than doxorubicin, and 24 h earlier than mitoxantrone. Caspase-3 inhibitor Ac-DEVD-CHO did not show any significant effect on drug cytotoxicity and DNA nucleosomal fragmentation. PMID- 16215685 TI - Characterization of the lipolytic pathways that mediate free fatty acid release during Fas/CD95-induced apoptosis. AB - We have undertaken a study to characterize the lipolytic pathway responsible for the generation of free fatty acids (FFA) during Fas/CD95-induced apoptosis in Jurkat cells. It was initially shown that the cellular lipid fraction that suffered the major quantitative decrease during Fas-induced apoptosis was that of phosphatidylcholine (PC). In addition, the secretion of palmitic acid-derived FFA was largely prevented by D609, an inhibitor of PC-specific phospholipase C (PC PLC) and also by the diacylglycerol lipase (DAGL) inhibitor RHC-80267, suggesting that the secretion of these FFA during Fas-induced apoptosis is mediated by the generation of DAG by a PC-PLC activity and, sequentially, by a 1-DAGL activity which generates the FFA from its sn-1 position. The endocannabinoid 2 arachidonoyl glycerol (2-AG) should be generated as a sub-product of this pathway, but it did not accumulate inside the cells nor was secreted into the supernatant. Interestingly, the complete inhibition of free AA secretion during Fas-induced apoptosis was only achieved by using the AA trifluoromethylketone, which not only inhibits all types of phospholipase-A(2) (PLA(2)) activities, but also the described lytic activities on 2-AG. Using a combination of RHC-80267 and the iPLA(2)-specific inhibitor bromoenol lactone, it was shown that the DAGL pathway also cooperates with iPLA(2) in the generation of free arachidonate. PMID- 16215686 TI - Nitrate tolerance aggravates postischemic myocardial apoptosis and impairs cardiac functional recovery after ischemia. AB - OBJECTIVES: This study examined the effects of nitrate tolerance (NT) on myocardial ischemia reperfusion (MI/R) injury and elucidated the potential mechanisms involved. Furthermore, the effects of GSH on postischemic myocardial apoptosis in NT rats were investigated. METHODS AND RESULTS: Male Sprague-Dawley rats were randomized to receive nitroglycerin (60 microg/kg/h) or saline for 12 h followed by 40 min of MI and 4 h of reperfusion. Myocardial apoptosis, infarct size, nitrotyrosine formation, plasma CK and LDH activity, and cardiac function were determined. MI/R resulted in significant apoptotic cell death, which was further increased in animals with NT. In addition, NT further increased plasma CK and LDH activity, enlarged infarct size, and impaired cardiac functional recovery after ischemia. Myocardial nitrotyrosine, a footprint for cytotoxic reactive nitrogen species formation, was further enhanced in the NT heart after MI/R. Treatment of NT animals with exogenous GSH inhibited nitrotyrosine formation, reduced apoptosis, decreased infarct size, and improved cardiac functional recovery. CONCLUSION: Our results demonstrate that nitrate tolerance markedly enhances MI/R injury and that increased peroxynitrite formation likely plays a role in this pathologic process. In addition, our results suggest that GSH could decrease peroxynitrite formation and reduce MI/R injury in nitrate tolerant hearts. PMID- 16215687 TI - Alkaline stress-induced apoptosis in human pulmonary artery endothelial cells. AB - The effect of alkaline stress, or an increase in extracellular pH (pHext), on cell viability is poorly defined. Human pulmonary artery endothelial cells (HPAEC) were subjected to alkaline stress using different methods of increasing pHext. Viability and mode of cell death following alkaline stress were determined by assessing nuclear morphology, ultrastructural features, and caspase-3 activity. Incubation of monolayers in media set to different pHext values (7.4 8.4) for 24-h induced morphological changes suggesting apoptosis (35-45% apoptotic cells) following severe alkaline stress. The magnitude of apoptosis was related to the severity of alkaline stress. These findings were confirmed with an assessment of ultrastructural changes and caspase-3 activation. While there was no difference in the intracellular calcium level ([Ca(2+)](i)) in monolayers set to pHext 7.4 versus 8.4 following the first hour of alkaline stress, blockade of calcium uptake with the chelator, EGTA, potentiated the magnitude of apoptosis under these conditions. Potentiation of apoptosis was reduced by calcium supplementation of the media. Finally, alkaline stress was associated with an increase in intracellular pH. This is the first report of apoptosis following alkaline stress in endothelial cells in the absence of other cell death stimuli. PMID- 16215688 TI - Differential gene expression during apoptosis induced by a serum factor: role of mitochondrial F0-F1 ATP synthase complex. AB - The number of genes that are up regulated or down regulated during apoptosis is large and still increasing. In an attempt to characterize differential gene expression during serum factor induced apoptosis in AK-5 cells (a rat histiocytoma), we found subunit 6 and subunit 8 of the transmembrane proton channel and subunit alpha of the catalytic core of the mitochondrial F(0)-F(1) ATP synthase complex to be up regulated during apoptosis. The increase in the expression levels of these subunits was concomitant with a transient increase in the intracellular ATP levels, suggesting that the increase in cellular ATP content is a result of the increase in the expression of ATP synthase subunits' gene and de novo protein synthesis. Depleting the cellular ATP levels with oligomycin inhibited apoptosis significantly, pointing to the requirement of ATP during apoptosis. Caspase 1 and caspase 3 activity and the loss of mitochondrial membrane potential were also inhibited by oligomycin during apoptosis in these cells, suggesting that the oligomycin induced inhibition of apoptosis could be due to inhibition of caspase activity and inhibition of mitochondrial depolarization. However, cytochrome C release during apoptosis was found to be completely independent of intracellular ATP content. Besides the ATP synthase complex genes, other mitochondrial genes like cytochrome C oxidase subunit II and III also showed elevated levels of expression during apoptosis. This kind of a mitochondrial gene expression profile suggests that in AK-5 cells, these genes are upregulated in a time-linked manner to ensure sufficient intracellular ATP levels and an efficient functioning of the mitochondrial respiratory chain for successful completion of the apoptotic pathway. PMID- 16215689 TI - Lycopene induces apoptosis in immortalized fibroblasts exposed to tobacco smoke condensate through arresting cell cycle and down-regulating cyclin D1, pAKT and pBad. AB - There is a lot of interest in the health benefits of dietary carotenoids and on the relationship of these compounds with smoke. In particular, it is unknown if the enhanced cancer risk observed in smokers following beta-carotene supplementation can be also found using other carotenoids. Here, we studied the effects of the tomato carotenoid lycopene on molecular pathways involved in cell cycle progression, apoptosis and survival in immortalized RAT-1 fibroblasts exposed to cigarette smoke condensate (TAR). Lycopene (0.5-2.0 microM) inhibited cell growth in a dose-and time-dependent manner, by arresting cell cycle progression and by promoting apoptosis in cells exposed to TAR. The arrest of cell cycle was independent of p53 and of 8-OH-dG DNA damage and related to a decreased expression of cyclin D1. Moreover, the carotenoid up-regulated apoptosis and down-regulated the phosphorylation of AKT and Bad in cells exposed to TAR. Such an effect was associated to an inhibition of TAR-induced expression of Cox-2 and hsp90, which is known to maintain AKT activity. This study suggests that lycopene, differently from beta-carotene, can exert protective effects against cigarette smoke condensate. PMID- 16215691 TI - The cell death machinery controlled by Bax and Bcl-XL is evolutionarily conserved in Ciona intestinalis. AB - Bax and Bcl-XL are key regulators of apoptosis in mammals. Here we report the functional characterization of two Bcl-2 homologues, ciBax and ciBcl-XL, in a basal invertebrate-chordate ascidian Ciona intestinalis. CiBax is a Ciona homologue of the BH1-3 pro-apoptotic protein Bax, whereas ciBcl-XL is a Bcl-XL like anti-apoptotic protein. Molecular modeling analysis showed that ciBax and ciBcl-XL share both sequence and structural similarities to human Bax and Bcl-XL, respectively. Like their human counterparts, ciBax could form a homodimer or oligomers as well as heterodimerize with ciBcl-XL, and overexpression of ciBax caused apoptosis that could be attenuated by ciBcl-XL. Mutagenesis studies showed that the BH3 domain of ciBax is critical for its cell death-inducing function and also for its interaction with ciBcl-XL. In Ciona embryos, ectopic expression of ciBax but not its BH3 deletion mutant resulted in cell dissociation and apoptosis after late gastrula stage of embryonic development. Moreover, not only wild type ciBcl-XL but also a mutant ciBcl-XL(F101V), which is unable to interact with ciBax, could block cell dissociation and developmental deficit in Ciona embryos induced by overexpression of ciBax. Taken together, these findings suggest that functional homologues of both the BH1-3 death effector Bax and the pro-survival Bcl-XL exist in sea squirt Ciona intestinalis, and they control the cell death machinery independent of their heterodimerization. PMID- 16215690 TI - Apoptosis in UV-C light irradiated p53 wild-type, apaf-1 and p53 knockout mouse embryonic fibroblasts: interplay of receptor and mitochondrial pathway. AB - Mouse embryonic fibroblasts (MEFs) deficient for the transcription factor p53 are hypersensitive to UV-C light. They also show a reduced recovery from UV-C induced replication blockage and are unable to repair UV-C photoproducts. In this study, we utilized wild-type (wt), Apaf-1 deficient (apaf-1(-/-)) and p53 deficient (p53(-/-)) MEFs in order to elucidate the role of non-repaired UV-C lesions in apoptotic signalling. Corresponding with the cellular sensitivity determined by the WST assay, p53(-/-) cells displayed the highest level of apoptosis, whereas wt cells showed moderate apoptosis after UV-C irradiation. Apaf1(-/-) cells were most resistant. In wt cells apoptosis was executed both via the mitochondrial and the receptor-mediated pathway, as shown by Bcl-2 decline, induction of fasR and activation of caspases-3,8,9. In apaf-1(-/-) (p53(+/+)) cells, the mitochondrial pathway was blocked downstream of Bcl-2, indicating that in this case apoptosis was mediated via the induction of fasR and caspase-3,8 activation. In p53 deficient cells, non-repaired UV-C induced DNA lesions triggered sustained up regulation of fas ligand (fasL) mRNA, which was not seen in wt and apaf-1(-/-) cells. Therefore, in p53(-/-) MEFs, the receptor/ligand triggered pathway appeared to be dominant. This was confirmed by significant reduction of apoptosis after DN-FADD transfection. As opposed to wt and apaf-1(-/-) cells, p53 deficient MEFs showed no induction of Fas receptor and no Bcl-2 decline. Nevertheless, the resulting caspase-8 and -3 activation was stronger compared to wt and apaf-1(-/-) cells. The data indicate that UV-C light activates in MEFs both the Fas (CD95, Apo-1) receptor and the mitochondrial damage pathways. In p53(-/-) cells, however, the high level of non-repaired DNA damage forces signalling by fasL upregulation, leading to enhanced UV-C-induced apoptosis. PMID- 16215692 TI - The "Generacion Diez" after-school program and Latino parent involvement with schools. AB - The current study examines associations between participation in after-school programs and change in Latino parent involvement with schools. Hierarchical linear regression analyses demonstrated that parents of children who had higher after-school program attendance rates were significantly more likely to report increases in the quality of relationships with their children's teachers, frequency of parent-teacher contact, and engagement with their children's schooling over a two-year period. However, greater home educator contacts were related to decreases in quality and quantity of parent-school involvement. A primary implication is that attendance in school-based after-school programs may draw parents into children's regular-day school context. Editors' Strategic Implications The authors illustrate the promising practice of using after-school programs to promote parent involvement and to help integrate the often disparate family and school contexts for Latino children. PMID- 16215693 TI - The impact of culturally relevant violence prevention models on school-age youth. AB - The Family and Community Violence Prevention (FCVP) Program was established in 1994 to address the escalation of youth violence among ethnic minorities. This federally funded program adapted the public health model and organized Family Life Centers throughout the country to serve youth who were considered to be at risk for violence and other abusive behaviors. The purpose of this three-year study, 1999-2002, was to determine the effectiveness of the FCVP Program's six component curriculum in reducing violence among participants. Results from posttest scores of 2,315 youth showed girls 12 and over to be most at risk for deviant behaviors; the program was most effective with boys under age 12. Academic performance and bonding to school were protective factors whereas exposure to violence was a risk factor for all four ethnic groups studied- African Americans, Hispanics, Native Americans, and Native Hawaiians. EDITORS' STRATEGIC IMPLICATIONS: Cultural anthropologists, public health specialists, and school officials should know that prevention programs can be designed to reflect the unique, culturally appropriate norms of specific ethnic minority groups, even as these programs address shared risk factors. The authors discuss the promising strategy of enhancing academic performance and school bonding to serve as protective factors against school violence, but they also describe age, gender, and cultural differences that must be addressed in future research. PMID- 16215694 TI - Proteomic analysis of cadmium-induced protein profile alterations from marine alga Nannochloropsis oculata. AB - Protein profile alterations following exposure to cadmium were examined in marine alga Nannochloropsis oculata through proteomic analysis. Alterations of the protein expression patterns following 10 muM cadmium treatment were analyzed on 2 dimensional gels. Out of 380 protein spots detected on 2-D gel using Coomassie staining, 11 spots were changed significantly following cadmium treatment. Because of the non-availability of molecular background information on this non sequenced algal species, cross-species protein identification through ESI-Q-TOF MS/MS was used to identify altered proteins. Two newly induced proteins were identified as malate dehydrogenase orthologue and NADH dehydrogenase orthologue. One suppressed protein was identified to be glyceraldehydes 3-phosphate dehydrogenase A. Protein spot showing a 3-fold increase was identified as mitochondrial NADH: ubiquinone oxidoreductase orthologue. However, we could not find any matches in the database from ESI-Q-TOF MS/MS for the remaining seven proteins, thus only partial peptide sequences of these proteins were found. PMID- 16215695 TI - Cholinesterases in aquatic biomonitoring: assay optimization and species-specific characterization for a California native fish. AB - Cholinesterase (ChE) enzyme activity measurements are widely applied in aquatic organisms for water quality monitoring, especially for pesticide contamination in agricultural watersheds. These biomarkers are amenable to measurement in a variety of species, and are therefore useful for examining effects in model organisms relevant to the ecosystem of interest. However, extensive variation in ChE biochemistry exists among tissues and species. This variation is rarely characterized and may lead to biases in the interpretation of activity determinations. We optimized ChE activity measurement parameters and characterized ChE biochemistry in Sacramento sucker (Catostomus occidentalis), a widely distributed fish native to watersheds of the Central Valley of California. Acetylcholinesterase (AChE) was the predominant ChE present in C. occidentalis brain and muscle, and muscle AChE was most sensitive to diazinon inhibition. Field caging experiments indicated that exposures to ChE-inhibiting pesticides were insufficient to induce neurotoxic effects. However, pesticide usage in the Central Valley is highly variable among years, and long-term monitoring of in stream effects would be necessary to evaluate trends in pesticide contamination. Recent changes to the State Water Code require agricultural landowners to participate in a regional water quality monitoring plan. As with most regional monitoring plans, measurements of in-stream effects, and effects in resident species, are not scheduled to be included. We suggest that inclusion of biomarker measures would lend important information to the monitoring process, and propose these procedures as a template for adapting ChE activity measurements into region specific monitoring programs to assess in-stream effects of pesticide contamination on native species. PMID- 16215696 TI - An assessment of PCBs and OC pesticides in eggs of double-crested (Phalacrocorax auritus) and Pelagic (P. pelagicus) cormorants from the west coast of Canada, 1970 to 2002. AB - Eggs of double-crested and pelagic cormorants were collected between 1970 and 2002 from colonies in the Strait of Georgia, BC, Canada, and assayed for concentrations of organochlorine (OC) pesticides and polychlorinated biphenyls (PCBs). Double-crested cormorant eggs from the early 1970's contained up to 4.1 mg kg-1 p,p'-DDE and 12.5 mg kg-1 sigmaPCBs. Corresponding values for pelagic cormorant eggs were 1.5 mg kg-1 p,p'-DDE and 3.9 mg kg-1 sigmaPCBs. Egg tissue concentrations of the dominant OC pesticides and sigmaPCBs dropped mainly during the 1970's, with minor declines thereafter. The data suggest that contaminant levels in cormorants have now stabilized at low levels throughout the resident population. Small but significant latitudinal gradients in several OC pesticides and PCBs indicated that areas of the southern strait were more contaminated than areas of the less populated northern strait. Interspecific differences in contamination may indicate that pelagic cormorants have a reduced capacity to metabolize chlordanes, DDT and PCBs compared to double-crested cormorants. Alternatively, the two species may have more divergent prey bases than previously thought. During the 1980's, TCDD toxic equivalents, largely contributed by polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), were correlated with physiological and biochemical alterations. Also, from 1989 to 1990, four deformed cormorant chicks (two of each species) were found during nest visits; none were found between 1991 and 1995. The relative rates of deformed chicks were 6 per 10,000 for double-crested and 16 per 10,000 for pelagic cormorants. The findings of deformed chicks were coincident with the period of highest PCDD and PCDF contamination; however, the sample sizes were too low to derive a substantive connection. Colony-wide productivity of double-crested cormorants was poorer in the southern colonies where PCBs in particular were elevated. While of concern, these lines of evidence are insufficient to conclude that chlorinated hydrocarbon contamination was a dominant contributor to population declines. It seems more probable that ecological variables, particularly changing prey and predator dynamics, drove the reductions in population size. PMID- 16215697 TI - Degradation of abamectin and doramectin on sheep grazed pasture. AB - Avermectins are widely used veterinary medicines. They bind strongly to faeces in their non-metabolized form and their half-life in faeces depends on field conditions. There are conflicting data regarding the behaviour of avermectins in the environment. Therefore, we investigated the degradation of abamectin and doramectin on sheep grazed pasture under field conditions in soil, soil-faeces and faeces samples from day 6 to day 70 (abamectin) or to day 50 (doramectin) after sheep treatment. Field conditions were recorded periodically during the experiment. Degradation of abamectin in sheep faeces and in soil-faeces was observed until day 60, with small amounts present in faeces until 70 days post treatment. Because the concentration of abamectin residues in soil was very low on day 6 after treatment, further significant degradation could not be measured. The concentration of doramectin in all analysed matrices decreased rapidly until day 50. It can be concluded that high concentrations of both avermectins were present during the first 20 days after treatment and that field conditions have an important role in degradation of avermectins on grazed pasture of treated animals. Clear identification of the consequences of avermectin exposure and the period of the greatest environmental risk will require further investigations. PMID- 16215698 TI - Fluctuating asymmetry and growth as biomarkers for exposure to androgen disrupting chemicals in Japanese quail. AB - The effects of embryonic exposure to androgen disrupting chemicals (ADCs) on growth and fluctuating asymmetry (FA) were determined in Japanese quail chicks. Embryos were exposed to an anti-androgenic chemical, 1,1,1-Trichloro-2,2-bis(p chlorophenyl)ethane (p,p'-DDE) at 20 or 40 microg, or to an androgenic chemical, trenbolone acetate, at 5 or 50 microg on day one of incubation. Growth was measured by body weight and tarsus and culmen lengths from day of hatch until day 29. FA was measured as differences in right versus left lengths of the tarsus, radius, zygomatic process, and premaxilla in day old carcasses. No differences in FA were observed for either treatment. Embryonic exposure to DDE resulted in no significant differences in all measures of growth, although the same quail exhibited significant differences in immunological, reproductive, and behavioral measurements (reported elsewhere). Chicks exposed to trenbolone exhibited no differences in body weight or measures of FA at day of hatch, however, subsequent growth was inhibited. This study shows that although growth and FA are often used as measures of chemical stress experienced during embryonic development, they are not sensitive measures for exposure to these ADCs at these levels in Japanese quail. PMID- 16215699 TI - A review of factors influencing measurements of decadal variations in metal contamination in San Francisco Bay, California. AB - This review summarizes some of the principal results of systematic measurements of trace metal concentrations throughout San Francisco Bay that began in 1989, and that have yielded insights on the factors controlling temporal and spatial variations of those concentrations on seasonal to decadal time scales. Pronounced seasonal variation in some metal concentrations is associated with gradients in the system's hydrology and the diagenetic remobilization of metals from benthic sediments. Additional temporal variation is associated with interannual differences in hydrologic flushing (e.g., ENSO cycles) and episodic storm events. While intra- and inter-annual variabilities complicate assessments of long-term variations in metal concentrations, recent analyses using stable lead isotopic composition distributions and time-series models have deconvoluted decadal changes in lead and silver concentrations in the estuary. Decadal variations in concentrations of other contaminant metals (e.g., mercury) are now being characterized, as well as projections of future concentrations of other metals of concern (e.g., copper). These historic assessments and projections of trace metal variations attest to the importance of long-term, systematic monitoring programs to quantify past and future impacts on water quality in San Francisco Bay and other complex estuarine systems. PMID- 16215700 TI - Effect of varying physicochemistry of European surface waters on the copper toxicity to the green alga Pseudokirchneriella subcapitata. AB - Most standard toxicity test results, used in present environmental risk assessment and water quality criteria (WQC) setting procedures are obtained with standard test media that are not representative for natural surface waters when metal toxicity modifying factors like pH, water hardness and dissolved organic carbon (DOC) are considered. The aim of this study was, using the green alga Pseudokirchneriella subcapitata, (1) to investigate the individual effects of Ca, Mg (the hardness cations) and pH on the toxicity of copper in reconstituted artificial test waters and (2) to study the copper toxicity in 13 spiked surface waters originating from different European eco-regions. Surface waters were selected such that a broad range of DOC (1.55-20.4 mg/l), pH (5.52-8.30) and water hardness (7-238 mg CaCO(3)/l) was covered. Tests in reconstituted artificial waters demonstrated that the 72 h-E(b)C50 (expressed as dissolved Cu) increased by about a factor of 3 when the Ca and Mg concentrations increased from 0.25 to 2.5 mM. When pH was increased from 5.8 to 8.0, dissolved 72 h-E(b)C50 decreased by a factor of 3. It is suggested that competition between Cu2+, Ca2+, Mg2+ and H+ ions at the cell surface are the most likely explanation for these observations. Dissolved 72 h-E(b)C50s in the natural surface waters varied between 32.0 and 245 mug Cu/l and were up to a factor 15 higher than the 72 h E(b)C50 in standard artificial medium (16.5+/-4.8 mug Cu/l). Consequently, Water Effect Ratio's (WER, the ratio between the EC50 in natural water to the EC50 in standard test water) ranged from 1.9 to 14.8. Linear regression analysis revealed that higher E(b)C50 were significantly related to higher DOC-concentration of the natural waters (R2 = 0.69), but that water hardness and pH did not show a significant relation with copper toxicity in these surface waters. In European surface waters, a positive correlation is observed between water hardness and pH. As a result, hardness and pH effects on copper toxicity are counteractive in European surface waters, resulting in the highly significant relation between the 72 h-E(b)C50 and DOC-concentration. Normalisation of the obtained effect concentrations using a Biotic Ligand based predictive Cu-toxicity model revealed that variation in DOC and pH are mainly responsible for the observed differences of Cu-toxicity in natural waters. PMID- 16215702 TI - On science, stories, quality and quantity. PMID- 16215701 TI - Antineural antibody in patients with Tourette's syndrome and their family members. AB - It has been proposed that antineural antibodies were present in patients with Tourette's syndrome (TS) and other neuropsychiatric disorders. The purpose of our study was to investigate the presence of antineural antibodies in the individuals with Tourette's syndrome and the family members of TS patients. The sera of four TS patients with no current streptococcal infection, their tic-free family members including father, mother and sibling, and a age-matched control group who were tic free were assayed for antineural antibodies directed against rat tissue and neurons in primary cell culture. There were prominent antineural antibodies present in TS patients and their first-degree family members, but not in the control group. Western blotting showed proteins of about 120 kDa in their sera that were not present in the sera of controls. The preliminary results of our study suggest the importance of genetic vulnerability in the immunological pathophysiology of tic disorders. Future studies should investigate the interactions of genetics, environment, infectious agents, and immunity on symptom expression in families with tic disorders. PMID- 16215706 TI - Radicular pain after vertebroplasty: complication and prevention. PMID- 16215705 TI - Mice conditionally lacking the Wolfram gene in pancreatic islet beta cells exhibit diabetes as a result of enhanced endoplasmic reticulum stress and apoptosis. AB - AIMS/HYPOTHESIS: Wolfram syndrome is an autosomal recessive disorder characterised by childhood diabetes mellitus, optic atrophy and severe neurodegeneration, resulting in premature death. The aim of this study was to investigate the mechanisms responsible for the phenotype of carbohydrate intolerance and loss of pancreatic beta cells in this disorder. MATERIALS AND METHODS: To study the role of the Wolfram gene (Wfs1) in beta cells, we developed a mouse model with conditional deletion of Wfs1 in beta cells by crossing floxed Wfs1 exon 8 animals with mice expressing Cre recombinase under the control of a rat insulin promoter (RIP2-Cre). Complementary experiments using RNA interference of Wfs1 expression were performed in mouse insulinoma (MIN6) cell lines (WfsKD). RESULTS: Male knockout mice (betaWfs(-/-)) began developing variable and progressive glucose intolerance and concomitant insulin deficiency, compared with littermate controls, by 12 weeks of age. Analysis of islets from betaWfs(-/-) mice revealed a reduction in beta cell mass, enhanced apoptosis, elevation of a marker of endoplasmic reticulum stress (immunoglobulin heavy chain-binding protein [BiP]), and dilated endoplasmic reticulum with decreased secretory granules by electron microscopy. WfsKD cell lines had significantly increased apoptosis and elevated expression of the genes encoding BiP and C/EBP-homologous protein (CHOP), two markers of endoplasmic reticulum stress. CONCLUSIONS/INTERPRETATION: These results indicate that (1) lack of expression of Wfs1 in beta cells was sufficient to result in the diabetes mellitus phenotype; (2) beta cell death occurred by an accelerated process of apoptosis; and (3) lack of Wfs1 was associated with dilated endoplasmic reticulum and increased markers of endoplasmic reticulum stress, which appears to be a significant contributor to the reduction in beta cell survival. PMID- 16215707 TI - Periprosthetic soft tissue recurrence of chondroblastoma after attempted en bloc excision from the proximal humerus. AB - A case of soft tissue recurrence of chondroblastoma after attempted en bloc excision and endoprosthetic replacement is described. This tumor in the proximal humerus recurred after initial curettage and was subsequently treated by attempted en bloc excision with positive microscopic margins. The patient then presented with a large soft tissue recurrence surrounding the endoprosthesis. This periprosthetic recurrence necessitated re-excision and revision of the endoprosthesis. Recurrence is not uncommon following curettage of chondroblastoma. However, less is known about soft tissue recurrence after en bloc resection of this tumor with positive margins. A subset of chondroblastoma may exist with more locally aggressive behavior. PMID- 16215708 TI - [Gleason grading: diagnostic criteria and clinical implications]. AB - Prostate cancer offers a wide spectrum ranging from clinically insignificant to aggressive and fatal disease. The Gleason grade is a powerful prognostic indicator and does influence treatment decision. Educational programs and websites are currently available to improve reproducibility and reliability of Gleason grading. A major problem in reporting of Gleason grading in needle biopsy is undergrading. The Gleason grade 3 is the lowest grade which can be assigned reliably in needle biopsies. The major prognostic shift is between Gleason grades 3 und 4 which is characterized by fusion of the glandular formations. Reporting the proportion of Gleason grades 4 and 5 in needle biopsies may be critical in terms of treatment decisions. The present review deals with diagnostic criteria of the Gleason grades and its clinical implications. PMID- 16215709 TI - [The role of PSA in diagnosis of prostate cancer and its recurrence]. AB - Prostate specific antigen is the most important tumor marker of prostate cancer. PSA, in conjunction with digital rectal examination, is the first-line clinical tool for detection of prostate cancer. To improve its specificity PSA-density, PSA-ratio (fPSA/tPSA), PSA-velocity, and complexed PSA have been introduced into clinical praxis. The treatment of lower stage disease in younger men has resulted in a longer period of post-treatment cancer surveillance. Biochemical recurrence is an early indicator for clinical disease recurrence. PSA doubling time allows to distinguish between local and systemic progression and is also a valid predictor for distant metastasis and death of disease. PMID- 16215710 TI - Intrusive growth of flax phloem fibers is of intercalary type. AB - Flax (Linum usitatissimum L.) phloem fibers elongate considerably during their development and intrude between existing cells. We questioned whether fiber elongation is caused by cell tip growth or intercalary growth. Cells with tip growth are characterized by having two specific zones of cytoplasm in the cell tip, one with vesicles and no large organelles at the very tip and one with various organelles amongst others longitudinally arranged cortical microtubules in the subapex. Such zones were not observed in elongating flax fibers. Instead, organelles moved into the very tip region, and cortical microtubules showed transversal and helical configurations as known for cells growing in intercalary way. In addition, pulse-chase experiments with Calcofluor White resulted in a spotted fluorescence in the cell wall all over the length of the fiber. Therefore, it is concluded that fiber elongation is not achieved by tip growth but by intercalary growth. The intrusively growing fiber is a coenocytic cell that has no plasmodesmata, making the fibers a symplastically isolated domain within the stem. PMID- 16215711 TI - [Severity of chronic back pain. Assessment with the Mainz Pain Staging System]. AB - BACKGROUND: Among the possible reasons for low back pain, psychological factors are getting more attention. Early assessment of the stage of chronicity is important for successful therapy. The aim of the present study was to analyse the applicability of the Mainz Pain Staging System (MPSS) for chronic back pain outpatients. METHODS: Each of 679 back pain outpatients were assigned to one of the three MPSS stages of chronic back pain severity. Pain intensity and duration, anxiety and depression, pain sensation, impression of control and muscular capacity (isometric power of lumbal extensor), number of clinical consultations, hospitalisation, and days of work inability were also recorded. RESULTS: Males and females showed levels of chronic pain increasing with age. Patients with high MPSS levels showed significantly higher mean scores for anxiety, depression, number of consultations, hospitalisation, and days of inability to work. Furthermore, the same patients showed lower impressions of control and muscular capacity. CONCLUSIONS: The present study confirms the significance and usefulness of the MPSS applied to outpatients with back pain. To assess completely the stage of chronicity, psychological and physical/functional parameters should also be obtained. PMID- 16215712 TI - Lack of effect of combination antibiotic therapy on mortality in patients with pneumococcal sepsis. AB - In order to determine whether combination antibiotic therapy decreases mortality after severe pneumococcal infection, a retrospective study of a cohort of 1,840 adult patients with severe sepsis or septic shock enrolled in two multicenter clinical trials between 1994 and 1999 was conducted. Among 107 patients with monobacterial pneumococcal sepsis, the case-fatality rate was 20% (five of 25) for patients who received antibiotic monotherapy compared with 19.5% (16 of 82) for patients who received combination therapy (adjusted hazard ratio, 1.1; 95% CI, 0.4-3.1). Similarly, monotherapy did not increase the risk of death (adjusted hazard ratio, 1.0; 95% CI, 0.2-4.8) among bacteremic patients (n=75). However, the latter analysis may have been underpowered (power, 58%) to detect a difference in mortality. Overall, in contrast to recently published reports, these results suggest that combination antibiotic therapy does not decrease mortality after severe pneumococcal sepsis. PMID- 16215714 TI - Isolation and characterization of a carotenoid oxygenase gene from Chlorella zofingiensis (Chlorophyta). AB - The green alga Chlorella zofingiensis produces large amounts of the valuable ketocarotenoid astaxanthin under dark, heterotrophic growth conditions, making it potentially employable for commercial production of astaxanthin as feed additives, colorants, and health products. Here, we report the identification and characterization of a beta-carotene oxygenase (CRTO) gene that is directly involved in the biosynthesis of ketocarotenoids in C. zofingiensis. The open reading frame of the crtO gene, which is interrupted by three introns of 243, 318, and 351 bp, respectively, encodes a polypeptide of 312 amino acid residues. Only one crtO gene was detected in the genome of C. zofingiensis. Furthermore, the expression of the crtO gene was transiently up-regulated upon glucose treatment. Functional complementation in Escherichia coli showed that the coding protein of the crtO gene not only exhibits normal CRTO activity by converting beta-carotene to canthaxanthin via echinenone, but also displays a high enzymatic activity of converting zeaxanthin to astaxanthin via adonixanthin. Based on the bifunctional CRTO, a predicted pathway for astaxanthin biosynthesis in C. zofingiensis is described, and the CRTO is termed as carotenoid 4,4'-beta-ionone ring oxygenase. PMID- 16215715 TI - Optimization of culture on the overproduction of TRAIL in high-cell-density culture by recombinant Escherichia coli. AB - Different nutrient-feeding cultures were carried out in producing recombinant protein of truncated tumor necrosis factor related apoptosis-inducing ligand (TRAIL) (114-281 amino acids of TRAIL) in Escherichia coli strain C600/pBV-TRAIL. The effects of preinduction specific growth rate, postinduction carbon source (glucose and glycerol), and feeding strategies were investigated. The higher preinduction specific growth rate (mu = 0.22 h(-1)) contributed to the increase in the TRAIL production, at which TRAIL was accumulated in bacterial cells as 7.2% of total cellular protein, corresponding to 1.99 g l(-1) in contrast with 5.1% (1.29 g l(-1)) at preinduction specific growth rate (mu = 0.1 h(-1)) during high-cell-density culture. Glycerol was superior to glucose as the postinduction carbon source for TRAIL production. Under similar culture conditions, the final concentration of TRAIL was produced 1.59-fold more when glycerol was used as postinduction carbon source than when glucose was used. At the same time, the results showed that it is efficient to adopt the pH-stat feeding strategy at postinduction for the overproduction of TRAIL. The TRAIL production was increased up to 4.51 g l(-1), approximately 16.1% of total cellular protein. The mechanisms behind the preinduction specific growth rate effect on the expression level may be ascribed to the leakage secretion of acetate. PMID- 16215716 TI - Single-step ion exchange purification of the coagulant protein from Moringa oleifera seed. AB - The coagulant protein from Moringa oleifera (MO) seed was purified using a single step batch ion exchange (IEX) method. Adsorption and elution parameters were optimized. Impact of the purification on the reduction of organic and nutrient release to the water was studied. The matrix was equilibrated using ammonium acetate buffer, and the optimum ionic strength of NaCl for elution was 0.6 M. The time for adsorption equilibrium was between 90 and 120 min. Maximum adsorption capacity of the matrix, estimated with the Langmuir model, was 68 mg protein/g adsorbent. The purified protein does not release organic and nutrient loads to the water, which are the main concerns of the crude extract. This work suggests that a readily scalable single-step IEX purification method can be used to produce the coagulant protein and it can be carried out with locally available facilities. This will promote the use of MO in large water treatment plants and other industries. PMID- 16215713 TI - Interaction between simvastatin and metoprolol with respect to cardiac beta adrenoceptor density, catecholamine levels and perioperative catecholamine requirements in cardiac surgery patients. AB - Beta-blockade is a standard cardiovascular therapy known to induce the up regulation of beta-adrenoceptor density. Upon ligand-binding, beta-adrenoceptors are normally internalised via the arrestin pathway, and after dissociation they are re-inserted into the membrane. This means that at high catecholamine levels the adrenoceptor density is low and under beta-blockade it is high. The insertion of receptors into the membrane is often dependent on farnesylation processes that can be inhibited by statins. We carried out a prospective, controlled, observational study to determine whether beta-blockade-induced up-regulation of beta-adrenoceptor density is attenuated by statin therapy and whether this would subsequently affect catecholamine consumption during surgery. We obtained pre operative blood samples and intra-operative biopsies of the right atrial appendage from 39 patients (age: 65+/-5 years; BMI: 28+/-1) undergoing coronary bypass surgery with or without simvastatin (20 mg/day) therapy and with or without concomitant metoprolol therapy (50 mg/day). The atrial tissue was used for radioligand-binding studies with (-)-[125I]-iodocyanopindolol (ICYP) and for assessment of the beta-adrenoceptor subtype distribution following standard protocols. In the blood samples, plasma adrenaline and noradrenaline concentrations were determined using HPLC. In all tissue samples, we found a total beta-adrenoceptor density of 38+/-4 fmol/mg protein in untreated controls; this which was up-regulated to 55+/-5 fmol/mg protein in patients receiving metoprolol. This increase in receptor number was nearly prevented completely by simvastatin therapy (42+/-5 fmol/mg protein). The up-regulation could be attributed to increases in the beta1-adrenoceptor subtype. In contrast, simvastatin alone had no effect on beta-adrenoceptor density. Pre-operative adrenaline levels were slightly reduced in all drug therapy groups (nonsignificant differences), while the levels noradrenaline were not significantly different among the groups. With respect to the perioperative catecholamine requirements, patients on metoprolol needed significantly less dopamine than control patients, while patients undergoing simvastatin/metoprolol therapy needed as much as the controls. The post-operative total catecholamine requirements were not different among the four groups of patients. There were no differences in plasma metoprolol concentration between patients receiving metoprolol alone and those receiving a combination of metoprolol and simvastatin. IN CONCLUSION: Simvastatin therapy seems to counter-regulate the up-regulation of beta-adrenoceptor density. In the up-regulated state induced by metoprolol therapy, the patients seemed to need less catecholamines during cardiac surgery, which may be due to the higher number of beta-adrenoceptors. Additional simvastatin therapy did not reduce post-operative catecholamine consumption. PMID- 16215717 TI - Photodynamic therapy-generated vaccine for cancer therapy. AB - A target tumor-derived whole cancer cell therapeutic vaccine was developed based on an in vitro pre-treatment by photodynamic therapy (PDT) and was investigated using a poorly immunogenic tumor model. The vaccine was produced by incubating in vitro expanded mouse squamous cell carcinoma SCCVII cells for 1 h with photosensitizer benzoporphyrin derivative (BPD), then exposing to light (690 nm, 1 J/cm2) and finally to a lethal X-ray dose. Treatment of established subcutaneous SCCVII tumors growing in syngeneic C3H/HeN mice with 2 x 10(7) PDT vaccine cells per mouse by a peritumoral injection produced a significant therapeutic effect, including growth retardation, regression and cures. Tumor specificity of this PDT-generated vaccine was demonstrated by its ineffectiveness when prepared from a mismatched tumor cell line. Vaccine cells retrieved from the treatment site at 1 h postinjection were intermixed with dendritic cells (DC), exhibited heat shock protein 70 on their surface, and were opsonized by complement C3. Tumor-draining lymph nodes treated by the PDT-vaccine contained dramatically increased numbers of DC as well as B and T lymphocytes (with enlarged memory phenotype fraction in the latter), while high levels of surface bound C3 were detectable on DC and to a lesser extent on B cells. The PDT-vaccine produced no therapeutic benefit against tumors growing in C3-deficient hosts. It is suggested that surface expression of heat shock proteins and complement opsonization are the two unique features of PDT-treated cells securing avid immune recognition of vaccinated tumor and the development of a strong and effective antitumor adaptive immune response. PMID- 16215718 TI - A phase II trial of vaccination with autologous, tumor-derived heat-shock protein peptide complexes Gp96, in combination with GM-CSF and interferon-alpha in metastatic melanoma patients. AB - The aim of this study was to determine the immunogenicity and antitumor activity of autologous, tumor-derived heat shock protein gp96-peptide complex vaccine (HSPPC-96; Oncophage given with GM-CSF and IFN-alpha in pre-treated metastatic (AJCC stage IV) melanoma patients. Patients underwent surgical resection of metastatic lesions for HSPPC-96 production. HSPPC-96 was administered subcutaneously (s.c.) in four weekly intervals (first cycle). Patients with more available vaccine and absence of progressive disease received four additional injections in 2-week intervals (second cycle) or more. GM-CSF was given s.c. at the same site at days -1, 0 and +1, while IFN-alpha (3 MU) was administered s.c. at a different site at days +4 and +6. Antigen-specific anti-melanoma T and NK lymphocyte response was assessed by enzyme-linked immunospot assay on peripheral blood mononuclear cells obtained before and after vaccination. Thirty-eight patients were enrolled, 20 received at least four injections (one cycle) of HSPPC 96 and were considered assessable. Toxicity was mild and most treatment-related adverse events were local erythema and induration at the injection site. Patients receiving at least four injections of HSPPC-96 were considered evaluable for clinical response: of the 18 patients with measurable disease post surgery, 11 showed stable disease (SD). The ELISPOT assay revealed an increased class I HLA restricted T and NK cell-mediated post-vaccination response in 5 out of 17 and 12 out of the 18 patients tested, respectively. Four of the five class I HLA restricted T cell responses fall in the group of SD patients. Vaccination with autologous HSPPC-96 together with GM-CSF and IFN-alpha is feasible and accompanied by mild local and systemic toxicity. Both tumor-specific T cell mediated and NK cell responses were generated in a proportion of patients. Clinical activity was limited to SD. However, both immunological and clinical responses were not improved as compared with those recorded in a previous study investigating HSPPC-96 monotherapy. PMID- 16215719 TI - Why were limbs amputated? An evaluation of 216 surgical specimens from Chiang Mai University Hospital, Thailand. AB - INTRODUCTION: Limb loss has a devastating effect on patients. To know the underlying causes of limb amputation would be helpful in planning public health strategies in the country. The objectives of this study are (1) to identify the primary causes and the feature of limb amputations in the setting of a university hospital, and (2) to study the time trends of the causes of limb amputation over a period of 5 years. MATERIALS AND METHODS: The clinical and pathological data from 216 amputated limbs submitted to the Pathology Department of Chiang Mai University Hospital from 2000 to 2004 were reviewed. RESULTS: Of these, 188 cases were first time amputations, and 28 cases were repeat amputations. The 188 first amputated specimens included 23 upper limbs (12%) and 165 lower limbs (88%), from 115 male (61%) and 73 female (39%) patients. Dysvascular (46%), tumor-related (36%), and infection-related (10%) amputations were the three most common scenarios. The rate of amputation was high in 2004 (32%) owing to an unexpected increase in the numbers of dysvascular amputation. Atherosclerosis accounted for at least 52% of dysvascular amputations. The leading cause of tumor-related amputations was sarcoma (72%), almost half of which were osteosarcomas. The major cause of lower limb amputation was dysvascular (51%) whereas that of upper limb amputation was tumor related (61%). Subgroup analysis of the major limb amputations revealed that 44% were tumor related, 39% were dysvascular, and 8% were infection-related causes. The proportion of major limb losses in the tumor related group (87%, 59/68) was significantly higher than those in the dysvascular group (62%, 53/86) (P = 0.001). In addition, the proportion of upper limb losses in the tumor-related group (21%, 14/68) was significantly greater than those in the dysvascular group (2%, 2/86), (P < 0.001). The causes of 28 repeat amputations were similar, i.e., dysvascular (61%), tumor related (29%), and infectious related (7%). CONCLUSION: (1) Atherosclerosis, a potentially preventable disease is responsible for the great proportion of limb losses in Northern Thailand; (2) the numbers of dysvascular amputation seem to be increasing; (3) tumor, especially sarcoma, is the most common cause of major limb amputations as well as upper limb loss. PMID- 16215721 TI - Painful nonunion of fracture of the entire posterior process of the talus: a case report. AB - We report a 14-year-old boy in whom isolated nonunion fracture of the posterior process of the talus. He underwent surgical repair with Herbert Whipple screw fixation and plaster immobilization. Osseous union was achieved 3 months after surgery, resulting in the resolution of symptoms and complete functional recovery. To our knowledge, the present report is the first to describe a successful outcome for surgical treatment of painful nonunion of fracture of the entire posterior process of the talus. PMID- 16215720 TI - Expandable vertebral body replacement in patients with thoracolumbar spine tumors. AB - INTRODUCTION: The objectives of surgical interventions for tumoral lesions of the spine include the establishment and improvement of tumor-related symptoms. Anterior tumor resection followed by reconstruction indicated if surgical treatment allowed a marginal removal of the tumor or could extend the individual survival rate in combination with adjuvant therapy options. Sufficient re stabilization depends on adequate anterior column reconstruction. The purpose of this retrospective study was to present our experiences and results after anterior tumor resection followed by reconstruction with the expandable vertebral body replacement device (VBR, Ulrich, Germany) based on clinical application over 4 degrees years. PATIENTS AND METHODS: We carried out an anterior tumor resection followed by reconstruction using an anterior extendable device in 32 patients with different spine tumors between 1996 and 2000. A retrospective evaluation was executed considering the patients medical records and radiological findings. Additionally, a clinical and radiological investigation of still living postoperative patients was carried out. RESULTS: The mean surgical time of all evaluated patients was 317.2 min. The average blood loss was 1,272.5 ml. According to the Tokuhashi score, patients with a postoperative survival time of at least 12 months demonstrated a score value > or = 9 points. According to our evaluated patients group metastatic lesions of the spine represented the largest group (78.1%). The average survival rate of this group amounted to 18.4 months postoperatively. Considering primary tumors the average survival rate at the time of last re-examination amounted to 34.8 months postoperatively. Preoperative neurological pathologies were present in 12 patients (Frankel stage C-D). During the postoperative monitoring period 58.3% of the patients demonstrated an improvement in initial neurological findings. There were no intraoperative complications or perioperative deaths. Implant dislocations were not observed. CONCLUSION: On account of the underlying, the anterior tumor resection with supplementary instrumentation represented a sufficient procedure in spinal tumor surgery. Adjuvant therapy can influence the postoperative survival period positively in addition to the surgical procedure. Following anterior tumor resection, extendable vertebral body replacements like the VBR device provide immediate spine stability by excellent defect adaptation. With regard to their intraoperative flexibility, expandable cages are more advantageous in contrast to non-expandable implants or bone grafts. PMID- 16215722 TI - Second generation of meniscus transplantation: in-vivo study with tissue engineered meniscus replacement. AB - INTRODUCTION: The options available after meniscus loss offer only limited chances for a long-term success. In the following experimental study, we investigated the effect of meniscus tissue engineering on properties of the collagen meniscus implant (CMI). METHODS: Autologous fibrochondrocytes, obtained per biopsy from adult Merino sheep (n=25), were released from the matrix, cultured in-vitro and seeded into CMI scaffolds (n=10, group 1). Following a 3 week in-vitro culture, the tissue engineered menisci were used for autologous transplantation. Macroscopical and histological evaluation were performed in comparison with non-seeded CMI controls (n=10, group 2) and with meniscus resected controls (n=5, group 3) after 3 weeks (each 1 animal group 1 and 2) and 3 months. RESULTS: The lameness score did not show any difference between the groups. Meniscus tissue was found in seven knee joints (group 1), in five knee joints (group 2) and in two knee joints (group 3). The size of the transplants reduced from 25.9+/-4.5 to 20.1+/-10.8 mm (group 1) and from 25.9+/-1.5 to 14.4+/ 12.5 mm (group 2). Histologically, enhanced vascularisation, accelerated scaffold re-modelling, higher content of extra-cellular matrix and lower cell number were noted in the pre-seeded menisci in comparison with non-seeded controls. Dense high-cellular fibrous scar tissue was found in two of five cases in the resection control group. CONCLUSION: Tissue engineering of meniscus with autologous fibrochondrocytes demonstrates a macroscopic and histological improvement of the transplants. However, further development of the methods, especially of the scaffold and of the cell-seeding procedure must prove the feasibility of this procedure for human applications. PMID- 16215723 TI - Ischaemic cholangiopathy and sickle cell disease. AB - We report a case of a 6-year-old girl of Afro-Caribbean origin, known to have sickle cell disease (SCD), with recurrent history of jaundice and abdominal pain. She was extensively investigated, including endoscopic retrograde cholangiopancreatography (ERCP), which revealed diffuse cholangiopathy of both extrahepatic and intrahepatic bile ducts. A pigtail stent was placed and balloon dilatation was performed for stricture of the extrahepatic duct. Since then, she remains well and asymptomatic. We suggest that cholangiopathy is the consequence of sickling in the end arteries of the biliary arterial tree. PMID- 16215724 TI - Renal amyloidosis due to pulmonary tuberculosis in a patient with Down syndrome. AB - A pulmonary-tuberculosis-related renal amyloidosis patient with Down syndrome, who presented with nephrotic syndrome, was studied. PMID- 16215726 TI - Microbial population response to changes of the operating conditions in a dynamic nutrient-removal sequencing batch reactor. AB - A nutrient-removal sequencing batch reactor operated with short anaerobic/aerobic cycles was subjected to different operating conditions, namely, cycle length, feeding pattern and feed composition. The changes in microbial population, as well as the contribution of microbial groups to the total nutrient removal, were estimated using the kinetic parameters obtained in this study. Denitrifying polyphosphate-accumulating organisms (DPAOs) were detected in the system, representing a fraction of 23% of phosphorus-accumulating organisms (PAOs). The results suggest that DPAOs and non-DPAOs are different microorganisms. The presence of nitrate in the feed stimulated DPAOs to predominate over non-DPAOs. Feeding the reactor with a mixture of organic substrates also stimulated DPAOs. Glycogen-accumulating organisms (GAOs) were likely to be present in the system and their development over PAOs was apparently favoured by increasing the aeration time and feeding during the aerobic phase. In contrast, the presence of propanoate in the feed apparently favoured PAOs over GAOs. PMID- 16215725 TI - A fatal case of ruptured giant coronary artery aneurysm. AB - A 5-year-old Japanese boy died because of a ruptured left coronary artery aneurysm (CAA). He was diagnosed as having Kawasaki disease (KD) on the 5th day from onset, with all of the principal signs. On the 7th day of illness, bilateral CAAs were already found via echocardiography, and he was treated with intravenous (IV) gamma globulin and oral ASA. However, the fever persisted and the CAA progressed rapidly. Echocardiography on the 12th illness day showed a giant (18 mm) left anterior descending (LAD) artery aneurysm. Oral propranolol and nifedipine were administered, in conjunction with warfarin/aspirin anti coagulation therapy. On the 13th day of illness, cardiac arrest developed abruptly, and, despite cardiopulmonary resuscitation (CPR), the patient remained unresponsive and died one hour later. The final pathological diagnosis was a ruptured LAD artery aneurysm and cardiac tamponade. Microscopic investigation of the ruptured vascular wall revealed marked neutrophilic infiltration, with fewer macrophages and lymphocytes. CAA ruptures are a very rare, but fatal, complication of KD. Based on a review of previous reports on CAA ruptures, we consider it useful to distinguish aneurysms which rapidly dilate and continue to expand beyond a diameter of 10 mm with ongoing vasculitis (these CAAs can be termed "super-giant") from the more common giant CAAs limited to a diameter of 8 or 9 mm, because a decision must be made as to whether to start intensive care or to intervene surgically, in order to ensure the survival of patients with such a potentially critical complication. PMID- 16215727 TI - A model of xylitol production by the yeast Candida mogii. AB - Production of xylitol from xylose in batch fermentations of Candida mogii ATCC 18364 is discussed in the presence of glucose as the cosubstrate. Various initial ratios of glucose and xylose concentrations are assessed for their impact on yield and rate of production of xylitol. Supplementation with glucose at the beginning of the fermentation increased the specific growth rate, biomass yield and volumetric productivity of xylitol compared with fermentation that used xylose as the sole carbon source. A mathematical model is developed for eventual use in predicting the product formation rate and yield. The model parameters were estimated from experimental observations, using a genetic algorithm. Batch fermentations, which were carried out with xylose alone and a mixture of xylose and glucose, were used to validate the model. The model fitted well with the experimental data of cell growth, substrate consumption and xylitol production. PMID- 16215728 TI - Dynamic kinetic resolution: alternative approach in optimizing S-ibuprofen production. AB - A lipase catalysed enantioselective hydrolysis process under in situ racemization of the remaining (R)-ibuprofen ester substrate with sodium hydroxide as the catalyst was developed for the production of S-ibuprofen from (R,S)-ibuprofen ester in isooctane. Detailed investigations on parameters study indicated that 0.5 M NaOH, addition of 20% (v/v) co-solvent (dimethyl sulphoxide), operating temperature of 45 degrees C, and 40 mmol/L substrate gave 86% conversion and 99.4% optical purity of S-ibuprofen in dynamic kinetic resolution. Meanwhile, in common enzymatic kinetic resolution process, only 42% conversion of the racemate and 93% enantiomeric excess of the product was obtained which are of lower values as compared to dynamic kinetic resolution. The S-ibuprofen produced during each process was evaluated and approximately 50% increment in concentration of S-acid product was produced when dynamic kinetic resolution was applied into the process. PMID- 16215730 TI - Physical and genetic mapping of amplified fragment length polymorphisms and the leaf rust resistance Lr3 gene on chromosome 6BL of wheat. AB - The Argentinian wheat cultivar Sinvalocho MA carries the Lr3 gene for leaf rust resistance on distal chromosome 6BL. In this cultivar, 33 spontaneous susceptible lines were isolated and cytogenetically characterized by C-banding. The analysis revealed deletions on chromosome 6BL in most lines. One line was nulli-6B, two lines were ditelo 6BS, two, three, and ten lines had long terminal deletions of 40, 30, and 20%, respectively, three lines showed very small terminal deletions, and one line had an intercalary deletion of 11%. Physical mapping of 55 amplified fragment length polymorphism (AFLP) markers detected differences between deletions and led to the division of 6BL into seven bins delimited by deletion breakpoints. The most distal bin, with a length smaller than 5% of 6BL, contained 22 AFLP markers and the Lr3 gene. Polymorphism for nine AFLPs between Sinvalocho MA and the rust leaf susceptible cultivar Gamma 6 was used to construct a linkage map of Lr3. This gene is at a genetic distance of 0.9 cM from a group of seven closely linked AFLPs. The location of the gene in a high recombinogenic region indicated a physical distance of approximately 1 Mb to the markers. PMID- 16215729 TI - A composite linkage map from two crosses for the species complex Picea mariana x Picea rubens and analysis of synteny with other Pinaceae. AB - Four individual linkage maps were constructed from two crosses for the species complex Picea mariana (Mill.) B.S.P. x Picea rubens Sarg in order to integrate their information into a composite map and to compare with other Pinaceae. For all individual linkage maps, 12 major linkage groups were recovered with 306 markers per map on average. Before building the composite linkage map, the common male parent between the two crosses made it possible to construct a reference linkage map to validate the relative position of homologous markers. The final composite map had a length of 2,319 cM (Haldane) and contained a total of 1,124 positioned markers, including 1,014 AFLPs, 3 RAPDs, 53 SSRs, and 54 ESTPs, assembled into 12 major linkage groups. Marker density of the composite map was statistically homogenous and was much higher (one marker every 2.1 cM) than that of the individual linkage maps (one marker every 5.7 to 7.1 cM). Synteny was well conserved between individual, reference, and composite linkage maps and 94% of homologous markers were colinear between the reference and composite maps. The combined information from the two crosses increased by about 24% the number of anchor markers compared to the information from any single cross. With a total number of 107 anchor markers (SSRs and ESTPs), the composite linkage map is a useful starting point for large-scale genome comparisons at the intergeneric level in the Pinaceae. Comparisons of this map with those in Pinus and Pseudotsuga allowed the identification of one breakdown in synteny where one linkage group homologous to both Picea and Pinus corresponded to two linkage groups in Pseudotsuga. Implications for the evolution of the Pinaceae genome are discussed. PMID- 16215731 TI - Evidence of the domestication history of flax (Linum usitatissimum L.) from genetic diversity of the sad2 locus. AB - A phylogenetic analysis was conducted on 34 alleles of 2.5 kb sized stearoyl-ACP desaturase II (sad2), obtained from 30 accessions of cultivated and pale flax (Linum spp.), to elucidate the history of flax domestication. The analysis supports a single domestication origin for extant cultivated flax. The phylogenetic evidence indicates that flax was first domesticated for oil, rather than fibre. The genetic diversity of the sad2 locus in cultivated flax is low when compared to that of the pale flax assayed. An absolute archaeological date could be applied to the synonymous substitution rate of sad2 in cultivated flax, yielding a high estimate of 1.60-1.71x10(-7) substitutions/site/year. The occurrence of nonsynonymous substitutions at conserved positions of the third exon in alleles from cultivated flax suggests that the locus may have been subjected to an artificial selection pressure. The elevated synonymous substitution rate is also compatible with a population expansion of flax since domestication, followed by a population decline in historic times. These findings provide new insight into flax domestication and are significant for the continuous exploration of the flax germplasm for utilization. PMID- 16215732 TI - High-throughput DNA typing of HLA-A, -B, -C, and -DRB1 loci by a PCR-SSOP-Luminex method in the Japanese population. AB - We have developed a new high-throughput, high-resolution genotyping method for the detection of alleles at the human leukocyte antigen (HLA)-A, -B, -C, and DRB1 loci by combining polymerase chain reaction (PCR) and sequence-specific oligonucleotide probes (SSOPs) protocols with the Luminex 100 xMAP flow cytometry dual-laser system to quantitate fluorescently labeled oligonucleotides attached to color-coded microbeads. In order to detect the HLA alleles with a frequency of more than 0.1% in the Japanese population, we created 48 oligonucleotide probes for the HLA-A locus, 61 for HLA-B, 34 for HLA-C, and 51 for HLA-DRB1. The accuracy of the PCR-SSOP-Luminex method was determined by comparing it to the nucleotide sequencing method after subcloning into the plasmid vector using 150 multinational control samples obtained from the International HLA DNA Exchange University of California Los Angeles. In addition, we performed the PCR-SSOP Luminex method for HLA allele typing on DNA samples collected from 1,018 Japanese volunteers. Overall, the genotyping method exhibited an accuracy of 85.91% for HLA-A, 85.03% for HLA-B, 97.32% for HLA-C, and 90.67% for HLA-DRB1 using 150 control samples, and 100% for HLA-A and -C, 99.90% for HLA-B, and 99.95% for HLA DRB1 in 1,018 Japanese samples. The PCR-SSOP-Luminex method provides a simple, accurate, and rapid approach toward multiplex genotyping of HLA alleles to the four-digit or higher level of resolution in the Japanese population. It takes only approximately 5 h from DNA extraction to the definition of HLA four-digit alleles at the HLA-A, HLA-B, HLA-C, and HLA-DRB1 loci for 96 samples when handled by a single typist. PMID- 16215733 TI - Analysis of the expressed heavy chain variable-region genes of Macaca fascicularis and isolation of monoclonal antibodies specific for the Ebola virus' soluble glycoprotein. AB - The cynomolgus macaque, Macaca fascicularis, is frequently used in immunological and other biomedical research as a model for man; understanding it's antibody repertoire is, therefore, of fundamental interest. The expressed variable-region gene repertoire of a single M. fascicularis, which was immune to the Ebola virus, was studied. Using 5' rapid amplification of cDNA ends with immunoglobulin (Ig)G specific primers, we obtained 30 clones encoding full-length variable, diversity, and joining domains. Similar to the human V(H) repertoire, the M. fascicularis repertoire utilized numerous immunoglobulin heavy variable (IGHV) gene fragments, with the V(H)3 (41%), V(H)4 (39%), and V(H)1 (14%) subgroups used more frequently than the V(H)5 (3.9%) or V(H)7 (1.7%) subgroups. Diverse immunoglobulin heavy joining (IGHJ) fragments also appeared to be utilized, including a putative homolog of JH5beta gene segment identified in the related species Macaca mulatta, Rhesus macaque, but not in humans. Although the diverse V region genes in the IgG antibody repertoire of M. fascicularis had likely undergone somatic hypermutations (SHMs), they nevertheless showed high nucleotide identity with the corresponding human germline genes, 80-89% for IGHV and 72-92% for IGHJ. M. fascicularis and human V(H) genes were also similar in other aspects: length of complementarity-determining regions and framework regions, and distribution of consensus sites for SHMs. Finally, we demonstrated that monoclonal antibodies (mAbs) specific for an Ebola protein could be obtained from M. fascicularis tissue samples by phage display technology. In summary, the study provides new insight into the M. fascicularis V region gene repertoire and further supports the idea that macaque-derived mAbs may be of therapeutic value to humans. PMID- 16215734 TI - Mycoplasma pneumoniae pneumonia: CT features in 16 patients. AB - The objective of this study was to assess the computed tomography (CT) features of Mycoplasma pneumoniae pneumonia. We retrospectively reviewed CT findings of 16 patients (M:F = 9:7, age range 1-74 years, median 9 years) with serologically proven Mycoplasma pneumoniae pneumonia and with chest CT scan available. Two distinctive patterns of CT features of M. pneumoniae pneumonia were noted between the paediatric (age < 18 years) and the adult (age > or = 18 years) groups. The pediatric group (n=11) showed lobar or segmental consolidation (100%) with frequent pleural effusion (82%) and regional lymphadenopathy (82%) and mild volume decrease of the involved lobe (73%), while four of the five adult patients showed diffuse and/or multifocal, centrilobular or peribronchovascular areas of ground-glass attenuation (80%) with a lobular distribution, and frequent thickening of interlobular septa (60%) and the bronchial walls (40%) were also detected at high-resolution CT. The CT finding of a lobar or segmental consolidation with a parapneumonic effusion seen in our children with M. pneumoniae pneumonia was similar to that of bacterial lobar pneumonia. In contrast, the CT findings noted in our adult patients consisted of a mixture of a bacterial bronchopneumonia pattern and a viral interstitial pneumonia pattern. PMID- 16215735 TI - Adrenal masses falsely diagnosed as adenomas on unenhanced and delayed contrast enhanced computed tomography: pathological correlation. AB - OBJECTIVES: To assess the accuracy of CT for the diagnosis of histologically confirmed adrenal adenoma and nonadenoma using CT numbers. MATERIALS AND METHODS: Our study included 91 adrenal masses in 83 patients; histopathological diagnoses were 45 adenomas, 31 pheochromocytomas, 6 hyperplasias, 4 metastasis, and 5 miscellaneous lesions. Unenhanced CT in 46 patients and unenhanced and delayed contrast-enhanced (DCE) CT in 37 patients were retrospectively reviewed to examine the correlation between CT findings and those on pathological examination and to obtain diagnostic accuracy. RESULTS: Sensitivity, specificity, and accuracy for adenoma were 40% (18/45), 91% (42/46), and 66% (60/91) with unenhanced CT, and 96% (24/25), 61% (11/18), and 81% (35/43) with DCE CT. Adrenal masses falsely diagnosed as adenoma on unenhanced CT included three hyperplasias and one endothelial cyst, and those falsely diagnosed as adenoma on DCE CT were five pheochromocytomas, one oncocytic cortical tumor, and one primary pigmented nodular adrenocortical dysplasia. Twenty-five lipid-poor adenomas were falsely diagnosed as nonadenomas on unenhanced CT and one degenerated adenoma both on unenhanced CT and on DCE CT. CONCLUSION: Diagnosing adenoma merely on CT numbers can lead to misdiagnosis. The lower specificity than expected is due to pheochromocytomas presenting as false positives. PMID- 16215736 TI - Assessment of left ventricular myocardial function using 16-slice multidetector row computed tomography: comparison with magnetic resonance imaging and echocardiography. AB - OBJECTIVE: To assess functional parameters using multidetector-row computed tomography (MDCT) and echocardiography and to compare the results with magnetic resonance imaging (MRI). MATERIALS AND METHODS: End-diastolic-volume (EDV), end systolic-volume (ESV), stroke-volume (SV), ejection-fraction (EF), and myocardial mass (MM) were calculated based on CT data sets from 52 patients. Echocardiography was performed in 24 of the 52 patients. The results from MDCT and echocardiography were compared with MRI. RESULTS: A strong correlation between MDCT and MRI (r=0.66-0.90) was found for all parameters. Echocardiography revealed a low or moderate correlation (0.05-0.59). Compared to MRI the average differences with MDCT were for EDV 15.1 ml, ESV 10.6 ml, SV 4.5 ml, EF 1.8%, and MM 8.2 g, for EDV determined by echocardiography 36.2 ml, ESV 6.8 ml, and EF 13.9%. Bland-Altman analysis revealed acceptable limits of agreement between MRI and MDCT. CONCLUSIONS: MDCT enables reliable quantification of left ventricular function. Echocardiography was found to have only a moderate agreement of functional parameters with MRI. PMID- 16215737 TI - Age-related changes in cortical bone mass: data from a German female cohort. AB - To describe data from digital radiogrammetry (DXR) in an unselected German female cohort over a wide age range. Using a retrospective study design we analyzed radiographs of the hand from 540 German women (aged 5-96 years) using an automated assessment of cortical thickness, metacarpal index (MCI), and estimated cortical bone mineral density (DXR-BMD) on digitized radiographs. Both hands were radiographed in 97 women. In this group DXR-BMD and cortical thickness were significantly higher in the right metacarpals while there was no significant difference in MCI. To study the association with age we differentiated young (<20 years), middle-aged (20-45 years), and an older patients (>45 years). In young women all parameters increased significantly with age in a linear fashion (r=0.8 for DXR-BMD, r=0.7 for MCI). In those aged 25-45 years DXR-BMD and MCI were highest (peak bone mass). In women aged 45 or older all parameters decreased with age in an almost linear fashion with an annual change ranging from 0.7% to 0.9%. Our results for an unselected German female cohort indicate that DXR is a reliable, widely available osteodensitometric technique based on the refinement of conventional radiogrammetry. These findings are comparable to those from other studies and represent a valid resource for clinical application and for comparisons with other ethnic groups. PMID- 16215738 TI - The germ-line-restricted chromosome in the zebra finch: recombination in females and elimination in males. AB - In the zebra finch (Taeniopygia guttata), there is a germ-line-restricted chromosome regularly present in males and females. A reexamination of male and female meiosis in the zebra finch showed that this element forms a euchromatic bivalent in oocytes, but it is always a single, heterochromatic element in spermatocytes. Immunostaining with anti-MLH1 showed that the bivalent in oocytes has two or three foci with a localized pattern, indicating the regular occurrence of recombination. In male meiosis, the single restricted chromosome forms an axis that contains the cohesin subunit SMC3, and the associated chromatin is densely packed until late pachytene. Electron microscopy of thin-sectioned seminiferous tubules shows that the restricted chromosome is eliminated in postmeiotic stages in the form of packed chromatin inside a micronucleus, visible in the cytoplasm of young spermatids. The selective condensation of the restricted chromosome during early meiotic prophase in males is interpreted as a strategy to avoid the triggering of asynaptic checkpoints, but this condensation is reversed prior to the final condensation that leads to its (ulterior) elimination. Recombination during female meiosis may prevent the genetic attrition of the restricted chromosome and, along with the elimination in male germ cells, ensures its regular transmission through females. PMID- 16215739 TI - Independent differentiation of mammotropes and somatotropes in the chicken embryonic pituitary gland. Analysis by cell distribution and attempt to detect somatomammotropes. AB - It has been reported that mammotropes in a rodent pituitary gland are derived from somatotropes via somatomammotropes (SMTs), cells that produce both growth hormone (GH) and prolactin (Prl). However, no studies have been done on the transdifferentiation of somatotropes in the chicken pituitary gland. In this study, in order to determine the origin of mammotropes, we studied detail property of appearance of chicken somatotropes, mammotropes and pit-1 cells and then evaluated the existence of SMTs in the chicken embryonic pituitary gland. Immunohistochemical analysis revealed that GH-immunopositive (GH-ip) cells appeared on embryonic day (E) 14 and were mainly distributed in the caudal lobe, while Prl-immunopositive (Prl-ip) cells appeared in the cephalic lobe of the pituitary gland on E16. In situ hybridization (ISH) and RT-PCR analysis showed that expression of GH and Prl mRNA starts at E12 in the caudal lobe and at E14 in the cephalic lobe respectively. Pit-1 mRNA was first detected on E5 by RT-PCR, and pit-1 mRNA-expressing cells were found in the cephalic lobe on E8. Then with the ontogeny of the chicken, these cells spread into both lobes. Using a double staining method with ISH and immunohistochemistry, we could not detect the existence of SMTs in the chicken embryonic pituitary gland even in the marginal region of either lobe. These results suggest that chicken somatotropes and mammotropes independently appear in different lobes of pituitary gland and that transdifferentiation from somatotropes to mammotropes is not the central route for differentiation of mammotropes in the embryonic chicken pituitary gland. PMID- 16215740 TI - Immunoelectron microscopic study of polyamines in the gastrointestinal tract of rat. AB - Polyamines (PAs) are ubiquitous polycationic metabolites in the eukaryotic and prokaryotic cells and are believed to be intimately involved in the regulation of DNA, RNA, and protein biosynthesis. However, the subcellular localization of PAs has not yet been fully elucidated in a variety of cell types. In the present study, a pre-embedding indirect immunoperoxidase approach was used to define the fine structural localization of PAs in the gastrointestinal tract of rat, which was fixed with glutaraldehyde and the monoclonal antibody ASPM-29 specific for spermine (Spm) and spermidine (Spd). Examination by a transmission electron microscopy showed that the peroxidase end products were commonly and predominantly localized in the free and attached ribosomes of the rough endoplasmic reticulum (rER) in the active protein- or peptide-secreting cells, and in rapidly proliferating cells including the gastric chief cells, mucous neck cells, and intestinal crypt cells. The nuclei, mitochondria, and secretory vesicles were devoid of PAs. Of note is the new finding that PAs are also located even on the small number of ribosomes in the cytoplasm of the parietal cells and of the villus-tip cells, because these were the cell types that were found to be almost PA-negative at the light microscopic level. These results seem to be completely consistent with those recently obtained for rat neurons. Thus, the present study generalized the subcellular localization of PAs on the ribosomes, and demonstrated that PAs are one of the components of biologically active ribosomes, possibly in any type of cell, that are closely involved in the translation processes of protein biosynthesis. PMID- 16215741 TI - Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. AB - In vitro studies on the pathogenesis in swine have been hampered by the lack of relevant porcine cell lines. Since many bacterial infections are swine-specific, studies on pathogenic mechanisms require appropriate cell lines of porcine origin. We have characterized the permanent porcine intestinal epithelial cell line, IPEC-J2, using a variety of methods in order to assess the usefulness of this cell line as an in vitro infection model. Electron microscopic analyses and histochemical staining revealed the cells to be enterocyte-like with microvilli, tight junctions and glycocalyx-bound mucin. The functional integrity of monolayers was determined by transepithelial electrical resistance (TEER) measurements. Both commensal bacteria and important bacterial pathogens were chosen for study based on their principally different infection mechanisms: obligate extracellular Escherichia coli, facultative intracellular Salmonella and obligate intracellular Chlamydia. We determined the colonization and proliferation of the bacteria on and within the host cells and monitored the host cell response. We verified the expression of mRNAs encoding the cytokines IL 1alpha, -6, -7, -8, -18, TNF-alpha and GM-CSF, but not TGF-beta or MCP-1. IL-8 protein expression was enhanced by Salmonella invasion. We conclude that the IPEC J2 cell line provides a relevant in vitro model system for porcine intestinal pathogen-host cell interactions. PMID- 16215742 TI - Histone lysine methylation patterns in human cell types are arranged in distinct three-dimensional nuclear zones. AB - The impact of histone lysine methylation as an essential epigenetic mechanism for gene regulation has been demonstrated by numerous studies where it was functionally and structurally linked to euchromatin and heterochromatin. Most of these data have been obtained by biochemical and two-dimensional (2D)-microscopic techniques providing little information about the global nuclear arrangement of histone modifications. We investigated the 3D architecture and spatial interrelationships of different histone lysine methylation sites (tri-H3K4, mono H4K20, mono-H3K9, tri-H3K27, tri-H4K20 and tri-H3K9) in various human cell types. Immunofluorescence and confocal microscopy were used together with a quantitative evaluation of 3D images, to reveal spatial relations of specific methylation sites with either centromeres, nascent RNA or with each other. A close association with centromeres was found only for histone methylation sites previously linked to constitutively repressed chromatin. Differences observed in these sites in relation to the cell cycle emphasize the potential relevance of the dynamic properties of heterochromatin for nuclear functions. Nascent RNA was found associated, though to a different degree, with all histone methylation sites, supporting the increasing evidence that transcription occurs across a wide range of the human genome. Finally we demonstrated by simultaneous visualization of different histone lysine methylation sites that methylation patterns are organized in distinct nuclear zones with little apparent intermingling. PMID- 16215743 TI - Collinearity, curvature interpolation, and the power of perceptual integration. AB - In three experiments, participants determined the orientation of a global triangle formed by three Gabor patches of a target spatial frequency in a field of distracters. The orientation of the target patches and their proximity were varied between conditions. When all the target patches had the same orientation this facilitated the response compared to random orientations. This effect occurred only when the patches were in close proximity. When the orientations of the target patches were different but aligned to the global triangle, facilitation occurred regardless of proximity. These contrasting types of facilitation were attributed to different early perceptual integration mechanisms that enable the perception of holistic structure. PMID- 16215744 TI - The psychology of film: perceiving beyond the cut. AB - First-order editing violations in film refer either to small displacements of the camera position or to small changes of the image size. Second-order editing violations follow from a reversal of the camera position (reversed-angle shot), leading to a change of the left-right position of the main actors (or objects) and a complete change of the background. With third-order editing violations, the linear sequence of actions in the narrative story is not obeyed. The present experiment focuses on the eye movements following a new shot with or without a reversed-angle camera position. The findings minimize the importance of editing rules which require perceptually smooth transitions between shots; there is also no evidence that changes in the left-right orientation of objects in the scene disturb the visual processing of successive shots. The observed eye movements are due either to the redirecting of attention to the most informative part on the scene or to attention shifts by motion transients in the shot. There is almost no evidence for confusion and/or for activities to restore the spatial arrangement following the reversal of the left-right positions. PMID- 16215745 TI - Emergent perceptual features in the benefit of consistent stimulus-response mappings on dual-task performance. AB - Duncan (1979) examined all combinations of compatible and incompatible stimulus response mappings for two spatial three-choice tasks in the psychological refractory period paradigm. Performance was better when the mappings for the tasks were consistent than when they were not, even when both mappings were incompatible. He attributed the benefit for the consistent incompatible mapping to an emergent choice between mappings when they are inconsistent that slows performance. Consistent incompatible mappings also may benefit from emergent perceptual features. The present study examined the role of emergent perceptual and mapping-choice features in two experiments that used pairs of two-choice tasks. Results similar to Duncan's were obtained with visual stimuli mapped to keypresses at short (stimulus onset asynchrony) SOAs. However, the benefit of the consistent incompatible mapping condition over the inconsistent mapping conditions was eliminated at an SOA of 1,000 ms. Furthermore, this benefit was not evident when the stimuli were auditory for Task 1 and visual for Task 2. With two-choice tasks, the benefit for consistent mappings apparently is due primarily to an emergent perceptual feature. PMID- 16215746 TI - Task-set inertia and memory-consolidation bottleneck in dual tasks. AB - Three dual-task experiments examined the influence of processing a briefly presented visual object for deferred verbal report on performance in an unrelated auditory-manual reaction time (RT) task. RT was increased at short stimulus-onset asynchronies (SOAs) relative to long SOAs, showing that memory consolidation processes can produce a functional processing bottleneck in dual-task performance. In addition, the experiments manipulated the spatial compatibility of the orientation of the visual object and the side of the speeded manual response. This cross-task compatibility produced relative RT benefits only when the instruction for the visual task emphasized overlap at the level of response codes across the task sets (Experiment 1). However, once the effective task set was in place, it continued to produce cross-task compatibility effects even in single-task situations ("ignore" trials in Experiment 2) and when instructions for the visual task did not explicitly require spatial coding of object orientation (Experiment 3). Taken together, the data suggest a considerable degree of task-set inertia in dual-task performance, which is also reinforced by finding costs of switching task sequences (e.g., AC --> BC vs. BC --> BC) in Experiment 3. PMID- 16215747 TI - How attentional focus on body sway affects postural control during quiet standing. AB - The purpose of this study was to investigate how attentional focus on body sway affects postural control during quiet standing. To address this issue, sixteen young healthy adults were asked to stand upright as immobile as possible on a force platform in both Control and Attention conditions. In the latter condition, participants were instructed to deliberately focus their attention on their body sways and to increase their active intervention into postural control. The critical analysis was focused on elementary motions computed from the centre of pressure (CoP) trajectories: (1) the vertical projection of the centre of gravity (CoG(v)) and (2) the difference between CoP and CoG(v) (CoP-CoG(v)). The former is recognised as an index of performance in this postural task, whilst the latter constitutes a fair expression of the ankle joint stiffness and is linked to the level of neuromuscular activity of the lower limb muscles required for controlling posture. A frequency-domain analysis showed increased amplitudes and frequencies of CoP-CoG(v) motions in the Attention relative to the Control condition, whereas non-significant changes were observed for the CoG(v) motions. Altogether, the present findings suggest that attentional focus on body sway, induced by the instructions, promoted the use of less automatic control process and hampered the efficiency for controlling posture during quiet standing. PMID- 16215748 TI - Psychobiological personality dimensions in two environmental-illness patient groups. AB - The aim of the present study was to investigate the psychobiological personality dimensions in two subgroups of patients with environmental illness (EI). Fifty nine patients, 34 women and 25 men (aged 32-69 years), were referred for symptoms allegedly caused by abnormal sensitivity to either dental fillings (DF; n=26) or electromagnetic fields (EMF; n=33). For the evaluation of personality, the Swedish 238-item version of the Temperament and Character Inventory (TCI) was used. Compared with a control group, the EMF group scored higher on the temperament dimension Persistence. The DF group scored higher on the TCI subscales Harm Avoidance (fatigability and asthenia) and Self-Directedness (self acceptance). Women scored higher than men did on the Novelty Seeking and Reward Dependence (RD) dimensions in the DF group and on RD in the control group, indicating an inherited gender difference. No differences were found between men and women in the EMF group. Our results indicate that the high level of persistence found in the EMF group and the high level of fatigability and asthenia in combination with high self-acceptance found in the DF group represent vulnerable personalities. No significant differences were found between the two patient groups, indicating that these groups are quite similar regarding personality. This vulnerability can be expressed as various mental and somatic symptoms, which can be interpreted as EI symptoms by the affected individual. PMID- 16215749 TI - Comparison of in vivo visual and computer-aided tooth shade determination. AB - The aim of this study was to evaluate the performance of shade-determining devices. For the daily practitioner, it is essential to know whether modern computer-aided shade selection is reliable in everyday life. So the question of how the clinical usability of these machines could be rated has to be clarified. In the following, three actual devices available in the market were compared using a human observer's perception. The SpectroShade device (MHT Optic Research AG, 8155 Niederhasli, Switzerland), the ShadeVision device (X-Rite Co., Grandville, USA) and the Digital Shade Guide DSG4 (A. Rieth, 73614 Schorndorf, Germany) were assessed with respect to their agreement with the color perception of three examiners looking at 57 test persons (six teeth each for a total of 342). Shades were reported in Vita Classical shades. It could be demonstrated that every single human examiner showed a significantly higher agreement value (human group on average 40.2%) when compared with the remaining five methods than each computer-aided tooth shade determination device. The devices reached on average only a value of 28.6%, whereas the X-Rite ShadeVision showed a significant better result (33.2%) than the MHT SpectroShade and Rieth DSG4 (27.0 and 25.7%). Identical shade results given by all three methods of a group (group of three devices and three humans) were found to be rather low for the computer aided devices (9.9%) compared with humans (36.7%). All six methods together agreed in 3.3% of the cases. It becomes evident that the methods-especially the computer-aided shade determination-are rather divided about the respective tooth color. Deficiencies of the instrumental as well as the visual detection become obvious. The best agreement level was performed by the human examiners. The best agreement of the evaluated devices was obtained-generally as well as among the human testers-by the X-Rite ShadeVision system, followed at a statistically significant distance by the MHT SpectroShade and the Rieth DSG4. The agreement among the examiner group was 52.9%, significantly better than that of each device compared to this group (31.3% on average). Color detection and its realization are very complex. As shown, in many cases, computer-aided color shade determination of natural teeth seems to not reflect human perception. PMID- 16215750 TI - Cell membrane fluidity related to electroporation and resealing. AB - In this paper, we report the results of a systematic attempt to relate the intrinsic plasma membrane fluidity of three different cell lines to their electroporation behaviour, which consists of reversible and irreversible electroporation. Apart from electroporation behaviour of given cell lines the time course required for membrane resealing was determined in order to distinguish the effect of resealing time from the cell's ability to survive given electric pulse parameters. Reversible, irreversible electroporation and membrane resealing were then related to cell membrane fluidity as determined by electron paramagnetic resonance spectroscopy and computer characterization of membrane domains. We found that cell membrane fluidity does not have significant effect on reversible electroporation although there is a tendency for the voltage required for reversible electroporation to increase with increased membrane fluidity. Cell membrane fluidity, however, may affect irreversible electroporation. Nevertheless, this effect, if present, is masked with different time courses of membrane resealing found for the different cell lines studied. The time course of cell membrane resealing itself could be related to the cell's ability to survive. PMID- 16215751 TI - Spectral broadening of the Soret band in myoglobin: an interpretation by the full spectrum of low-frequency modes from a normal modes analysis. AB - In this work the temperature dependence of the Soret band line shape in carbon monoxy myoglobin is re-analyzed by using both the full correlator approach in the time domain and the frequency domain approach. The new analyses exploit the full density of vibrational states of carbon-monoxy myoglobin available from normal modes analysis, and avoid the artificial division of the entire set of vibrational modes coupled to the Soret transition into "high-frequency" and "low frequency" subsets; the frequency domain analysis, however, makes use of the so called short-times approximation, while the time domain one avoids it. Time domain and frequency domain analyses give very similar results, thus supporting the applicability of the short-times approximation to the analysis of hemeprotein spectra; in particular, they clearly indicate the presence of spectral heterogeneity in the Soret band of carbon-monoxy myoglobin. The analyses also show that a temperature dependence of the Gaussian width parameter steeper than the hyperbolic cotangent law predicted by the Einstein harmonic oscillator and/or a temperature dependence of inhomogeneous broadening are not sufficient to obtain quantitative information on the magnitude of an-harmonic contributions to the iron-heme plane motion. However, the dependence of the previous two quantities may be used to obtain semiquantitative information on the overall coupling of the Soret transition to the low-frequency modes and therefore on the dynamic properties of the heme pocket in different states of the protein. PMID- 16215752 TI - Intracellular redox state: towards quantitative description. AB - Redox state is a widely used term for the description of redox phenomena in biological systems. The regulating mechanisms responsible for maintaining the redox state are not yet fully known. But it was shown that changes in the redox state might lead to a cascade of intracellular events, beneficial or deleterious to the cell. There are several methods for the description of the intracellular redox state. These methods are based on using measured intracellular concentrations of reduced and oxidized glutathione in the Nernst equation. However, glutathione is not always a basic redox component in biological fluids, organelles, cells or tissues. As a result, changes in the intracellular redox state are not always accompanied by considerable changes of glutathione concentration. In this work it was proposed to use the concept of effective reduction potential for the quantitative characteristic of the intracellular redox state. The effective reduction potential was substantiated on the basis of a thermodynamic description. A new equation for the calculation of the effective reduction potential was derived. This equation summarizes the contribution of different oxidizing and reducing agents in the formation of an effective redox potential. The theoretical estimation of the effective reduction potential values for the different biological fluids and cells was carried out with the use of a method developed. PMID- 16215755 TI - Long-term development of the radionuclide exposure of murine rodent populations in Belarus after the Chernobyl accident. AB - As a determinant of the associated health risks, the behavior of radionuclides in natural ecosystems needs to be better understood. Therefore, the activity concentration of various long-lived radionuclides released due to the Chernobyl accident, and the corresponding contributions to the whole-body dose rate, was studied as a function of time in mammalian indicator species inhabiting the natural forest ecosystems of Belarus, the bank vole (Clethrionomys glareolus) and the yellow-necked mouse (Apodemus flavicollus). The activity concentrations of 137Cs, 134Cs, 90Sr, 238Pu, 239,240Pu, 241Pu and 241Am in soil and in animals were measured at five monitoring sites with different ground deposition of radionuclides at different distances from the destroyed reactor. The observed temporal pattern of the radionuclide activity concentration in the studied animal populations reflects the changes in biological availability of these isotopes for biota, mostly due to fuel particle destruction and appearance of dissolved and exchangeable forms of radionuclides. The time course of 134+137Cs activity concentrations in animal populations appeared as a sequence of increase, peak and decrease. Maximal levels of radiocesium occurred 1-2 years after deposition, followed by an exponential decrease. Concentrations of incorporated 90Sr increased up to the tenth year after deposition. The activity concentrations of transuranic elements (238Pu, 239,240Pu, 241Pu and 241Am) were much lower than those of the other radionuclides, in the studied animals. A considerable activity of 241Am in animals from areas with high levels of contamination was firstly detected 5 years after deposition, it increased up to the tenth year and is expected to increase further in the future. Maximal values of the whole-body absorbed dose rates occurred during the year of deposition, followed by a decrease in the subsequent period. Generally, this decrease was monotonic, mainly determined by the decrease of the external gamma-ray dose rate, but there were exceptions due to the delayed maximum of internal exposure. The inter-individual distributions of radionuclide concentrations and lifetime whole-body absorbed doses were asymmetric and close to log-normal, including concentrations and doses considerably higher than the population mean values. PMID- 16215754 TI - [The complement system and its possible role in the pathogenesis of age-related macular degeneration (AMD)]. AB - The discovery of the complement factor H (CFH) polymorphism in age-related macular degeneration (AMD) strongly suggests a causative role of the complement system in the pathogenesis of this disease. The complement system is part of the innate immune system and is closely associated with the cellular response and the adaptive immune system. This article provides an overview of the complement system and, taking the new data into account, of possible immunopathogenetic processes in AMD. PMID- 16215756 TI - Economic bismuth-film microsensor for anodic stripping analysis of trace heavy metals using differential pulse voltammetry. AB - Stripping analysis has been widely recognised as a powerful tool in trace metal analysis. Its remarkable sensitivity is attributed to the combination of a preconcentration step coupled with pulse measurements that generate an extremely high signal-to-background ratio. Mercury-based electrodes have traditionally been used to achieve high reproducibility and sensitivity in the stripping technique. Because of the toxicity of mercury, however, new alternative electrode materials are highly desired, particularly for on-site monitoring. Use of thin films of bismuth deposited on platinum or glassy-carbon substrates has recently been proposed as a possible alternative to mercury--bismuth is "environmentally friendly", of low toxicity, and is in widespread pharmaceutical use. In this paper the preparation of economic bismuth-film microelectrodes by electrodeposition on a copper substrate and their application to heavy metal analysis are described. Bismuth-film electrodes were prepared by potentiostatic electrodeposition. Optimum conditions for chemical and electrochemical deposition to obtain an adherent, reproducible, and robust deposit were determined. The suitability of such microelectrodes for analysis of heavy metals was evaluated by anodic stripping voltammetry of cadmium. The analytical performance of bismuth film electrodes for anodic stripping voltammetry of heavy metals was evaluated for non-deaerated solutions containing Cd2+, Pb2+, and Zn2+ ions. Well-defined peaks with low background current were obtained by use of differential pulse voltammetry. Linear calibration plots were obtained for Cd2+ in acidified tap water at concentrations ranging from 2 x 10(-8) to 1 x 10(-7) mol L(-1) and from 1 x 10(-7) to 1 x 10(-6) mol L(-1) with relative standard deviations of 5% (n = 15) at the 1 x 10(-7) mol L(-1) level. The method was then successfully used to monitor the Cd2+ content of plant extracts and validated by polarographic and ICP MS measurements. These results open the possibility of using bismuth-coated copper electrodes as an alternative to mercury-based electrodes for analysis of heavy metals. The main problem remaining, which prevents on-site monitoring of heavy metals, is the need to use slightly acidic media, because formation of bismuth hydroxide on the film surface above pH 4.3 leads to non-reproducible measurements. Further experiments will be performed to discover whether electrode conditioning can be used to enable reproducible measurement in on-site monitoring of cadmium in natural waters. Moreover, further study should be conducted to evaluate the potential of BiFE for analysis of several pollutants of interest that are usually determined electrochemically by using mercury-based electrodes. PMID- 16215757 TI - PPAR-gamma is expressed and NF-kB pathway is activated and correlates positively with COX-2 expression in stromal myofibroblasts surrounding colon adenocarcinomas. AB - PURPOSE: Accumulated evidence indicates that carcinogenesis is closely associated with the transformation of normal stroma into a 'reactive' stromal phenotype. The present study investigated the role of PPARgamma, COX-2 and p-IkB-alpha- important molecular targets of colon cancer chemoprevention--in this stromal remodeling by evaluating and comparing the expression of these factors in stromal myofibroblasts, macrophages and endothelial cells that surround normal colonic mucosa and colon cancer. METHODS: Immunohistochemical methodology was employed on archived paraffin-embedded sections prepared from tumors and adjacent normal colon from 45 patients with colon adenocarcinomas. Double immunostaining with the universal marker for myofibroblasts (alpha-smooth muscle actin/alpha-SMA) as second primary antibody was also performed. RESULTS: Stromal macrophages and endothelial cells expressed these factors both in normal colonic mucosa and colon cancer. By contrast, stromal myofibroblasts expressed PPARgamma, COX-2 and p-IkB alpha only in colon adenocarcinomas (77.7%, 100% and 100% of cases, respectively) and not in normal colon. COX-2 and p-IkB-alpha expressions were strongly correlated in these cells (P < 0.001). PPARgamma, COX-2 and p-IkB-alpha expression did not correlate with the stage or differentiation of the adenocarcinomas. CONCLUSIONS: NF-kB pathway is activated and COX-2 expression is upregulated in stromal myofibroblasts surrounding colon adenocarcinomas compared to normal colon. Induction of COX-2 expression is primarily induced by NF-kB. NSAIDs, selective COX-2 inhibitors and PPARgamma ligands may exert their chemoprophylactic properties through direct actions on these cells. PMID- 16215760 TI - Pentane-1,5-diol as a percutaneous absorption enhancer. AB - Propylene glycol (propane-1,2-diol) is the only diol widely used in dermatology. Pentane-1,5-diol is mainly used as a plasticizer in cellulose products and adhesives, in dental composites and in brake fluid compositions and as a preservative for grain. However, pentane-1,5-diol is also an effective solvent, water-binding substance, antimicrobial agent and preservative and may therefore replace several ingredients in a skin composition. The release of tri iodothyroacetic acid (TRIAC) and percutaneous absorption of hydrocortisone and mometasone furoate with either pentane-1,5-diol or propane-1,2-diol and 2-methyl pentane-2,4-diol (hexylene glycol), respectively, as enhancers was compared. The release of TRIAC was 21% higher when pentane-1,5-diol was used as an enhancer instead of propane-1,2-diol. The percutaneous absorption of hydrocortisone through the skin was increased 12 times with propane-1,2-diol compared to 4.4 times with pentane-1,5-diol. However, the percutaneous absorption of hydrocortisone into the skin was 50% higher with pentane-1,5-diol compared to propane-1,2-diol. There was no significant difference, between the original mometasone furoate cream, with 2-methyl-pentane-2,4-diol, and the new cream with pentane-1,5-diol in the amount of mometasone furoate that was absorbed into the skin and through the skin. However, the cosmetic properties of the new mometasone furoate cream was superior to the original mometasone furoate cream, for examples, no bad odour, more even texture, goes better into the skin and has less greasiness. Pentane-1,5-diol can be used as a technology platform, which adds a series of desirable properties to dermatological preparations and enhances product usability. This will result in improved formulations for a series of major and commonly used dermatological drugs. When used in pharmaceutical topical preparations, pentane-1,5-diol will increase the percutaneous absorption of the active substance and it is an efficient antimicrobial agent that will act as an effective preservative in topical formulations. Pentane-1,5-diol is cosmetically attractive, has low risk for skin and eye irritation compared to other diols, low toxicity risk and no bad odour. PMID- 16215759 TI - Early zygote-specific nuclease in mitochondria of the true slime mold Physarum polycephalum. AB - The active, selective digestion of mtDNA from one parent is a possible molecular mechanism for the uniparental inheritance of mtDNA. In Physarum polycephalum, mtDNA is packed by DNA-binding protein Glom, which packs mtDNA into rod-shaped mt nucleoids. After the mating, mtDNA from one parent is selectively digested, and the Glom began to disperse. Dispersed Glom was retained for at least 6 h after mtDNA digestion, but disappeared completely by about 12 h after mixing two strains. We identified two novel nucleases using DNA zymography with native-PAGE and SDS-PAGE. One is a Ca2+-dependent, high-molecular-weight nuclease complex (about 670 kDa), and the other is a Mn2+-dependent, high-molecular-weight nuclease complex (440-670 kDa); the activity of the latter was detected as a Mn2+ dependent, 13-kDa DNase band on SDS-PAGE. All mitochondria isolated from myxamoebae had mt-nucleoids, whereas half of the mitochondria isolated from the zygotes at 12 h after mixing had lost the mt-nucleoids. The activity of the Mn2+ dependent nuclease in the isolated mitochondria was detected at least 8 h after mixing of two strains. The timing and localization of the Mn2+-dependent DNase activity matched the selective digestion of mtDNA. PMID- 16215761 TI - SCCA2-transfected human keratinocytes show increased secretion of IL-1alpha and IL-6, but not of TNF-alpha. PMID- 16215762 TI - Acute skin alterations following ultraviolet radiation investigated by optical coherence tomography and histology. AB - Optical coherence tomography (OCT) appears to be a promising technique to study skin in vivo. As part of an exploratory study to investigate UV induced effects non-invasively we aimed to evaluate the kinetics of acute UVB- as well as UVA1 induced skin alterations by means of OCT, and to correlate the results obtained with routine histology. Twelve healthy subjects received daily 60 J/cm2 of UVA1 and 1.5 minimal erythema doses of UVB on their upper back over three consecutive days. One day (24 h) after the last UV exposure, OCT measurements and skin biopsies were performed in four subjects (day 1) on the centre of the irradiated sites and an adjacent non-irradiated control site. The same procedure was performed in four subjects 3 days and 6 days after irradiation, respectively. Prior to OCT assessment two waterproof marks were drawn on the centre of UVB and UVA1 exposed sites and the control site. The OCT scanner, SkinDex 300, was used in the RI1D measurement modus in order to investigate morphological features, epidermal thickness, and scattering coefficients. Immediately after OCT assessment, 4 mm punch biopsies were taken from the previously marked sites. OCT as well as histological examinations performed on day 1, 3, and 6, revealed markedly higher values for epidermal thickness on UVB exposed skin sites, and slightly increased epidermal thickening in UVA1 exposed sites. UVB exposed sites showed disruption of the entrance signal in the B-scan of OCT resulting in a thickened layer with a signal-poor centre corresponding to hyperkeratosis and parakeratosis as confirmed by routine histology. Surprisingly, the mean scattering coefficients of the epidermis were slightly lower on UVA1 exposed sites, as compared to non-irradiated skin. By contrast, the scattering coefficient of the upper dermis of UVA1 irradiated skin was hardly altered. Moreover, the scattering coefficient of the upper dermis assessed on UVB exposed skin on day 1 was clearly smaller than the scattering coefficient observed on non irradiated and UVA1 exposed skin. Conclusively, it was possible to demonstrate by means of OCT differences of epidermal thickness and pathological features of the stratum corneum following UV exposure. UVA1 induced epidermal pigmentation as well as UVB induced dermal inflammation may affect the light attenuation in the tissue indicated by a decrease of the scattering coefficient. OCT seems to be a useful tool to monitor UV induced effects in vivo. PMID- 16215758 TI - Unexplained complexity of the mitochondrial genome and transcriptome in kinetoplastid flagellates. AB - Kinetoplastids are flagellated protozoans, whose members include the pathogens Trypanosoma brucei, T. cruzi and Leishmania species, that are considered among the earliest diverging eukaryotes with a mitochondrion. This organelle has become famous because of its many unusual properties, which are unique to the order Kinetoplastida, including an extensive kinetoplast DNA network and U insertion/deletion type RNA editing of its mitochondrial transcripts. In the last decade, considerable progress has been made in elucidating the complex machinery of RNA editing. Moreover, our understanding of the structure and replication of kinetoplast DNA has also dramatically improved. Much less however, is known, about the developmental regulation of RNA editing, its integration with other RNA maturation processes, stability of mitochondrial mRNAs, or evolution of the editing process itself. Yet the profusion of genomic data recently made available by sequencing consortia, in combination with methods of reverse genetics, hold promise in understanding the complexity of this exciting organelle, knowledge of which may enable us to fight these often medically important protozoans. PMID- 16215763 TI - The changes in expression of ICAM-3, Ki-67, PCNA, and CD31 in psoriatic lesions before and after methotrexate treatment. AB - Although the effectiveness of methotrexate (MTX) in the treatment of psoriasis is very well established, the mechanism of action is poorly understood. It was suggested that the therapeutic effect of MTX in psoriasis might be mediated by inhibition of adhesion molecule expression. The aim of our study was to investigate the different effects of MTX treatment on cell proliferation, inflammatory infiltrate, adhesion molecules, and angiogenesis in psoriasis, and to clarify the mechanism by which MTX exerts its therapeutic effects. Clinical response, the morpho-phenotypic changes, epidermal thickness, and mitosis count were analyzed and the expression of CD31 and ICAM-3, proliferative markers such as Ki-67, PCNA, were evaluated by immunohistochemical techniques in lesional psoriatic epidermis, before and after the treatment with MTX in ten patients. In posttreatment biopsies a decrease in the degree of epidermal hyperplasia and a significant reduction in the severity of the inflammatory infiltrate (P<0.05) were observed. In addition, CD31 and ICAM-3 expression was significantly decreased on dermal cellular infiltrate, (respectively; P<0.05, P<0.01). Ki67 and PCNA expression were suppressed concurrently in about 90% of cases (P<0.01). We suggest that MTX may have an inhibitory effect on an initial integral component of the pathways that lead to psoriasis. Immunopharmacologic intervention in adhesion event has the potential to improve psoriasis. Inhibition of revascularization may be another mechanism of action of MTX. PMID- 16215765 TI - Identification of two novel mutations in Chinese patients with Dyschromatosis symmetrica hereditaria. AB - Dyschromatosis symmetrica hereditaria (DSH) is a rare autosomal dominant cutaneous disorder characterized by a mixture of hyperpigmented and hypopigmented macules of various sizes on the extremities. Pathogenic mutations in the DSRAD gene have recently been identified. In this study, we report and identify the mutations of the DSRAD gene in two Chinese pedigrees with DSH. Two novel mutations in the functional domains of the DSRAD gene were identified and verified in two pedigrees. The c.3244A>G (H1075R) mutation was found in all patients but not in the healthy individuals from family A and c.3335_3336delAT (Y1112fs-->1112X) mutation was found in three patients but not in the healthy family members from family B. Our data suggests that these two novel mutations in the DSRAD gene could cause DSH and add new variants to the repertoire of DSRAD mutations in DSH. PMID- 16215764 TI - Levels of matrix metalloproteinase-2, -9 and -8 in the skin, serum and saliva of smokers and non-smokers. AB - Smoking induces skin ageing, affects wound healing and inflammatory responses in skin and mucous membranes but the mechanisms behind these adverse effects of smoking are not clear. The objective was to elucidate the mechanisms of smoking related tissue damage, by comparing the levels of matrix metalloproteinases (MMPs) -2, -9, and -8 in the skin, serum and saliva of smokers and non-smokers. The study population consisted of 47 current smokers and 51 non-smokers, all males of Finnish origin. Skin samples from the upper inner arm were frozen in liquid nitrogen. Levels of MMP-2 and MMP-9 protein in the skin were assessed by zymography and MMP-8 isoforms were determined by Western blotting. From the serum samples, MMP-2 and MMP-9 were assessed by zymography and MMP-8 levels by time resolved immunofluorometric assay (IFMA). From the salivary samples, MMP-8 levels were analysed by IFMA and MMP-9 levels by capture activity assay. In skin tissue, lower levels of both the pro and active forms of MMP-9 and of the active forms of MMP-8 were found in the smokers compared to the non-smokers. In serum, higher levels of proMMP-2 and proMMP-9 were found in the smokers compared to the non smokers (P=0.001 and P<0.001, respectively), whereas MMP-8 levels did not differ significantly between the groups. Active forms of MMP-9 and MMP-2 could not be found in serum. In saliva, the amount of total MMP-9 was significantly lower in the smokers (156.0 U/ml) compared to the non-smokers (223.9 U/ml, P=0.032), whereas the levels of MMP-8 or active MMP-9 did not differ significantly between the groups. We conclude that smoking alters the levels of matrix metalloproteinases in skin tissue, serum and saliva, which may affect the turnover of extracellular matrix of skin even though the clinical impact of our findings is not clear. PMID- 16215766 TI - Arsenic removal from the aqueous system using plant biomass: a bioremedial approach. AB - Metal species released into the environment by technological activities tend to persist indefinitely, circulating and eventually accumulating throughout the food chain, thus becoming a serious threat to the environment. Environment pollution by toxic metals occurs globally through military, industrial, and agricultural processes and waste disposal. Bioremediation processes are the target of recent research and are considered low-cost, ecofriendly methods to alleviate the current problems of water decontamination, particularly for remote and rural areas. The present piece of work reports the unexploited sorption properties of the powdered seed of the plant Moringa oleifera (SMOS) for the removal of Arsenic [As(III) and As(V)] from aqueous solutions. Sorption studies, using standard practices, result in the standardization of optimum conditions such as biomass dosages (2.0 g), metal concentrations (25 ppm), contact time (60 min) and volume of the test solutions (200 ml) at pH 7.5, for As(III) and pH 2.5 for As(V). Maximum sorption for As(III) and As(V) species is 60.21 and 85.6%, respectively. Protein/Amino acid-Arsenic interactions are found to play an important role in the biosorption process using plant biomass SMOS. PMID- 16215767 TI - Antiangiogenic therapy for a large splenic hemangioma. AB - Hemangiomas involving the spleen are rare and seldom symptomatic. Treatment options for large lesions usually consist of splenectomy, embolization, or both. Antiangiogenic treatment has not been reported previously as an effective alternative for this type of lesion. We report our experience of successfully using glucocorticoids in an infant with a large hemangioma of the spleen. PMID- 16215768 TI - Inhibitors of proprotein convertases. AB - The discovery of mammalian subtilases, proprotein convertases (PCs) or subtilisin like proprotein convertases (SPCs), in 1990 was a result of sustained efforts in searching for enzyme/s responsible for maturation of inactive protein precursors. Since then, seven PCs have so far been discovered that cleave at the carboxy terminal of a basic amino acid characterized by the consensus sequence Arg/Lys/His-X-X/Lys/Arg-Arg downward arrow, where X denotes any amino acid other than Cys. Two additional PC subtypes--called subtilisin kexin isozyme 1 (SKI-1) or site 1 protease (S1P) and neural apoptosis regulated convertase 1 (NARC-1), also known as PCSK9--that cleave at the carboxy terminus of nonbasic amino acids were discovered later. Numerous studies revealed various important functional roles of PCs in health and diseases such as tumorigenesis, diabetes, viral infections, bacterial pathogenesis, atherosclerosis, and neurodegenarative diseases such as Alzheimer's. Owing to these findings, PCs became a promising frontier for treatment of diverse pathologies. Thus modulation of PC activity with designed inhibitors is an attractive proposition not only for intervention of diseases, but also for biochemical characterization of these enzymes. Various physiological and bioengineered proteins as well as small molecules such as peptide, peptidomimetic, and nonpeptide compounds as inhibitors of PCs have been described in the literature. Among the strategies used for design of PC inhibitors, the most successful is the one based on bioengineered serpin proteins, of which the best example is alpha1-PDX, the double mutant variant of alpha1-antitrypsin (from A(355)IPM(358) to R(355)IPR(358)). Others include small peptide inhibitors with C-terminal carboxyl function modified with a potent neucleophile or those containing pseudo or isosteric peptide bond at the scissile site of a suitable peptide substrate. Among nonpeptide PC inhibitors, the number is very limited. So far, these include 20-carbon atoms containing alicyclic diterpenes of andrographolide family and heterocyclic compounds that are ligands of Zn2+ and Cu2+ ions. Overall, these molecules display only a modest enzyme inhibition; however, they may serve as important lead structures for further development of more potent and specific nonpeptide PC inhibitors as potential therapeutic agents. Many PC inhibitors display their functional properties in proliferation, fertilization, tumorigenesis, obesity, embryogenesis, or diabetes via their inhibitory action on PC activities. PMID- 16215769 TI - Effects of elevated plasma adrenaline levels on substrate metabolism, effort perception and muscle activation during low-to-moderate intensity exercise. AB - The aim of this study was to differentiate the role of raised plasma adrenaline (Adr) concentrations from sympathoadrenal activation associated with moderate intensity exercise, on muscle activation, cardiopulmonary responses, fuel metabolism, and ratings of perceived exertion (RPE) during low-intensity exercise. Two groups of subjects (MOD, n=6; LOW, n=7) cycled on two occasions for 90 min. MOD cycled at 68% VO(2max) with saline infusion, and at 34% VO(2max) with Adr infusion. LOW cycled twice at 34% VO(2max), with either Adr or saline infusion. Infusions (0.015 g Adr/kg/min) started at 15 min and increased plasma [Adr] somewhat higher than during exercise at 68% VO(2max) (approximately 1.9 vs. 1.4 nM, at 75 min). Mean plasma glucose and lactate concentrations during LOW were significantly higher with Adr than saline infusion (5.1+/-0.6 vs. 4.4+/-0.3 mmol/l, P<0.01 and 2.1+/-0.8 vs. 1.3+/-0.5 mmol/l, P<0.01, respectively). Elevated [Adr], without increased exercise intensity, did not alter glycogenolysis. There were also no effects of Adr infusion at 34% VO(2max) on heart rate, oxygen consumption, [FFA], respiratory exchange ratio, intramuscular triglyceride utilization, muscle activation or RPE. In conclusion, elevated [Adr] similar to those found during moderate-intensity exercise increased plasma glucose and lactate availability, but did not alter intramuscular fuel utilization, effort perception or muscle activation. PMID- 16215770 TI - [Pathophysiology of acne]. AB - Several pathogenic factors contribute to the development of acne, among them, seborrhea, follicular hyperkeratosis, propionibacteria, and inflammatory events. This article reviews current knowledge of these pathogenic factors. PMID- 16215773 TI - Tepidimonas taiwanensis sp. nov., a novel alkaline-protease-producing bacterium isolated from a hot spring. AB - The bacterial strain designated I1-1(T) was isolated from a hot spring located in the Pingtung area, southern Taiwan. Cells of this organism were Gram reaction negative rods, motile by a single polar flagellum. Optimum conditions for growth were 55 degrees C and pH 7. Strain I1-1(T) grew well in lower nutrient media such as 5-10% Luria-Bertani broth, and its extracellular products expressed alkaline protease activity. The 16S rRNA gene sequence analysis indicates that strain I1 1(T) is a member of beta-Proteobacteria. On the basis of a phylogenetic analysis of 16S rDNA sequences, DNA-DNA similarity data, whole-cell protein analysis, physiological and biochemical characteristics, as well as fatty acid compositions, the organism belonged to the genus Tepidimonas and represented a novel species within this genus. The predominant cellular fatty acids of strain I1-1(T) were 16:0 (about 41%), 18:1 omega7c (about 13%), and summed feature 3 [16:1 omega7c or 15:0 iso 2OH or both (about 26%)]. Its DNA base ratio was 68.1 mol%. We propose to classify strain I1-1(T) (=BCRC 17406(T)=LMG 22826(T)) as Tepidimonas taiwanensis sp. nov. PMID- 16215775 TI - Viscoelastic properties of Carbopol 940 gels and their relationships to piroxicam diffusion coefficients in gel bases. AB - PURPOSE: This study was conducted to determine the effect of formula compositions on viscoelastic properties of piroxicam gels using Carbopol 940 as a gelling agent and to determine the relationships between viscoelastic properties of Carbopol 940 gel bases and diffusion coefficients of piroxicam in gel bases. METHODS: Piroxicam gels (1.0% w/w) were prepared by using Carbopol 940 as a gelling agent and varying Carbopol 940 concentrations, glycerin, and sodium chloride contents. The in vitro release of piroxicam from gel bases to the receiving media, isotonic phosphate buffer solution (pH 7.4), were carried out using Franz-modified cell. The piroxicam diffusion coefficients were obtained by Higuchi's equation. Rheological property measurements of gel samples were performed via a cone and plate fluid rheometer. Relationships between viscoelastic properties of gel samples and piroxicam diffusion in gel bases were analyzed by Pearson's test at a p value of less than 0.05. RESULTS: All piroxicam gels exhibited predominantly elastic solid behavior whose magnitude depended on Carbopol 940 concentration. Preparations containing good solvent exhibited more elastic solid characters. In contrast, the piroxicam gels containing higher sodium chloride contents possessed more viscous fluid behavior. Analyzed by Pearson's test at a p value of less than 0.05, piroxicam diffusion coefficients were directly proportional to loss tangent, but were inversely proportional to storage modulus, loss modulus, complex modulus, and viscosity. CONCLUSIONS: There is a potential for predicting drug diffusion coefficients from their correlations to rheological parameters. This could be beneficial to the formulation design of transdermal drug delivery systems including mucoadhesive drug delivery systems. PMID- 16215774 TI - Additional materials of Myanmarpithecus yarshensis (Amphipithecidae, Primates) from the middle Eocene Pondaung Formation. AB - Myanmarpithecus yarshensis is an amphipithecid primate from the middle Eocene Pondaung Formation in Myanmar. It was previously known based on maxillary fragments with P(4)-M(3) and mandibular fragments with C-P(3) and M(2-3). This study reports new materials for the genus, including a humeral head fragment, a lingual fragment of the right M(2), a lingual fragment of the right M(3), and a left I(1). These new materials were collected from approximately the same point, and likely belonged to the same individual. The upper molar morphology and size of the new materials show similarity to those of the type specimen, indicating that the new materials can be assigned to M. yarshensis. The humeral head is the first postcranial element that is associated with dental materials for amphipithecids. The morphological similarity between the previously reported larger humerus and this specimen confirms the assignment of the former specimen to Amphipithecidae and suggests common locomotor adaptations in the family. The upper central incisor is large relative to the molar fragments, but is within the variation among extant platyrrhines. The tooth is spatulate-shaped and high crowned, and lacks the mesial process, indicating similarity to I(1) of haplorhines and clear differences from that of adapoids. It has been suggested that amphipithecids, including Myanmarpithecus, have affinities with notharctine adapoids, but the morphology of I(1) does not support the notharctine hypothesis of the Amphipithecidae. PMID- 16215792 TI - Employing patient expertise: introduction to the theme. PMID- 16215793 TI - The expert patient: outline of UK government paper. AB - This introduction outlines key elements in a recent United Kingdom Department of Health report that, it is hoped, will change attitudes, expectations and practices in the care of patients with chronic illness [Department of Health: 2001, The Expert Patient: A New Approach to Chronic Disease Management for the 21st Century. London: Department of Health.]. The findings of the Task Force are summarised as accurately as possible and without comment. Analysis and comment can be found in the accompanying papers. PMID- 16215794 TI - An expert in what?: the need to clarify meaning and expectations in "The Expert Patient". AB - This paper critiques particular aspects of the published UK government Department of Health's proposal to promote 'The Expert Patient' as a way of enhancing patient autonomy and reducing reliance on limited health care resources. Although the broad aims of the report are supported the detail is criticised on the basis that lack of clarity over key terms, including 'expert' 'illness' and 'disease', means that there is no clear focus for action and threatens to undermine the effectiveness of the proposals. PMID- 16215795 TI - Justifying patient self-management--evidence based medicine or the primacy of the first person perspective. AB - Patient self-management programs have become increasingly popular and are now also receiving official endorsements. This paper analyses two possible types of positive justifications for promoting patient self-management: evidence-based and patient-centred justifications. It is argued that evidence-based justifications, although important politically are deficient and that the primary justification for patient self-management must be a patient-centred justification focusing on the patient's privileged access to his or her own lived body. PMID- 16215796 TI - The expert patient: Illness as practice. AB - This paper responds to the Expert Patient initiative by questioning its over reliance on instrumental forms of reasoning. It will be suggested that expertise of the patient suffering from chronic illness should not be exclusively seen in terms of a model of technical knowledge derived from the natural sciences, but should rather include an awareness of the hermeneutic skills that the patient needs in order to make sense of their illness and the impact that the illness has upon their sense of self-identity. By appealing to MacIntyre's concepts of "virtue" and "practice", as well as Frank's notion of the "wounded story-teller", it will be argued that chronic illness can be constituted as a practice, by building a culture of honest and courageous story-telling about the experience of chronic suffering. The building of such a practice will renew the cultural resources available to the patient, the physician and the rest of the community in understanding illness and patient-hood. PMID- 16215797 TI - The expert patient: valid recognition or false hope? AB - The United Kingdom Department of Health initiative on "The Expert Patient" (2001) reflects recent trends in political philosophy, ethics and health services research. The overall objective of the initiative is to encourage patients, particularly those suffering from chronic conditions to become more actively involved in decisions concerning their treatment. In doing so there would be (perhaps) an expectation of better patient compliance and (arguably) a resultant improvement in quality of life. Despite these anticipated beneficial influences on health outcomes, there may be a danger that such initiatives are being carried along by the general swell of enthusiasm for recognising and facilitating the claims of disadvantaged or discriminated against groups. What more attractive than that patients should be "liberated" from what might be seen as the oppression of medical paternalism? To a great extent the potential for success of the Expert Patient venture turns on:(a) whether and to what extent a patient can be considered truly to be an expert and (b) full acceptance by the medical and heath care professions of allowing patients a more equitable and positive role. Whilst clearly the patient is an expert in the hermeneutic sense - it is they and they alone who experience their illness - there is nevertheless a risk of confusing experience with expertise. Experience limited to an individual does not of itself give rise to the generalisations that underlie reliable clinical treatment. Neither do the vast majority of patients possess the physiological and pharmacological knowledge to fully appreciate the biological nature of their illness nor the basis, risks or limitations of therapeutic measures. Might the notion of "The Expert Patient" as informed co-decision maker become a well meaning but rather vacuous aspiration similar to that of informed consent? Even worse, could patient "empowerment" have a deleterious effect? The paper reviews some of the major issues and concludes that the expert patient initiative could have benefits for both patients and health professionals if operated on the basis of concordance: an informed collaborative alliance that optimises the potential benefits of medical care. PMID- 16215798 TI - Bodily integrity and male and female circumcision. AB - This paper explores the ambiguous notion of bodily integrity, focusing on male and female circumcision. In the empirical part of the study we describe and analyse the various meanings that are given to the notion of bodily integrity by people in their daily lives. In the philosophical part we distinguish (1) between a person-oriented and a body-oriented approach and (2) between four levels of interpretation, i.e. bodily integrity conceived of as a biological wholeness, an experiential wholeness, an intact wholeness, and as an inviolable wholeness. We argue that bodily integrity is a prima facie principle in its own right, closely connected with, but still fundamentally different from, the principle of personal autonomy, that is, autonomy over the body. PMID- 16215799 TI - The story of the body and the story of the person: towards an ethics of representing human bodies and body-parts. AB - Western culture has a few traditions of representing the human body - among them mortuary art (gisants), the freak show, the culture of the relics, renaissance art and pre-modern and modern anatomy. A historical analysis in the spirit of Norbert Elias is offered with regard to body - person relationship in anatomy. Modern anatomy is characterized by separating the story of the person from the story of the body, a strategy that is incompatible with the bio-psycho-social paradigm of clinical medicine. The paper discusses different aspects of the above traditions and how they might bear on this conflict and on contemporary bioethics and bedside practice. PMID- 16215800 TI - The medical theory of Richard Koch I: theory of science and ethics. AB - Richard Koch first made his appearance in the 1920s with works published on the foundations of medicine. These publications describe the character of medicine as an action and the status of medicine within the theory of science. One of his conclusions is that medicine is not a science in the original sense of the word, but a practical discipline. It serves a practical purpose: to heal the sick. All medical knowledge is oriented towards this purpose, which also defines the physician's role. One kind of knowledge is diagnosis, which is strictly understood in relation to therapy, and is at the core of medical thinking. Diagnosis is not the assignment of a term of a species to a patient's disease: this would not do justice to the individuality of a clinical manifestation and would fail to provide a reason for individual therapy. Nevertheless, the terms assigned to diseases, although fictitious, are not useless, but assist in differentiating various phenomena. These conclusions carry ethical consequences. Because the task of helping the sick constitutes medicine, morals not only set ethical limits: medicine originates in a moral decision. If there are no diseases but only individual sick people, disease can not be defined as an abnormality. The individual benefit to the patient must not necessarily be the complete restoration of health. With its object being incalculable, medicine cannot guarantee its own success. Here the physician has to develop principles that allow for the best possible response to the challenges faced in varying situations of conduct. PMID- 16215801 TI - Health and autonomy. AB - Individual autonomy is a prominent value in Western medicine and medical ethics, and there it is often accepted that the only way to pay proper respect to autonomy is to let the patients themselves determine what is good for them. Adopting this approach has, however, given rise to some unwanted results, thus motivating a quest for an objective conception of health. Unfortunately, the purportedly objective conceptions of health have failed in objectivity, and if a conception of health is not acceptable for all agents, the threat of offending the patients' autonomy arises. This article sketches an objective conception of health that is able to respect individual autonomy. PMID- 16215802 TI - Respect for persons, respect for integrity: remarks for the conceptualization of integrity in social ethics. AB - Even though respect for integrity is hailed in several authoritative legal and ethical documents, and is typically presented as a complement to respect for autonomy, it is largely neglected in many leading works in ethics. Is such neglect warranted, or does it express a prejudice? This article argues that the latter is the case, and that this is due to misplaced conceptual concerns. It offers some proposals as regards the conceptualization of integrity in social ethics in general and in biomedical ethics in particular. Five main directions of interpretation of "integrity" are discerned and shown to be relevant for different areas of biomedical ethics. The defense of respect for integrity is served by a softening of principlism and by greater attention to context among the initial critics of this principle. PMID- 16215803 TI - Respect for persons, autonomy and palliative care. AB - This paper explores some of the values that underpin health care and how these relate more specifically to the values and ethics of palliative care. The paper focuses on the concept of autonomy because autonomy has emerged as a foundational concept in contemporary health care ethics and because this is an opportunity to scratch the surface of this concept in order to reveal something of its complexity, a necessary precaution when applying the concept to the context of palliative care. The paper begins with a theoretical discussion of autonomy exploring an aspect of its contemporary meaning and relevance to health care. The second part of the paper focuses more closely on how the principle of respect for autonomy can be applied in the context of palliative care. In this section an ethical framework is employed to explore a practical application of this principle within a broader context of respect for persons. PMID- 16215804 TI - The challenges of evidence-based medicine: a philosophical perspective. AB - Although evidence-based medicine (EBM) has gained prominence in current medical practice and research, it has also had to deal with a number of problems and inconsistencies. For example, how do clinicians reconcile discordant results of randomized trials or how do they apply results of randomized trials to individual patients? In an attempt to examine such problems in a structured way, this essay describes EBM within a philosophical framework of science. Using this approach, some of the problems and challenges faced by EBM can be explained at a more fundamental level. As well, by employing a similar description of the competing alternative research tradition of clinical medicine, this essay not only highlights the philosophical differences between these two modes of medical practice, but suggests that they, in fact, play a de facto complementary role in current clinical medicine. PMID- 16215810 TI - Metastatic tumors of the nervous system. PMID- 16215811 TI - Brain metastases: epidemiology and pathophysiology. AB - Metastases are the most common tumors of the central nervous system (CNS), but cancer databases are often incomplete leading to underestimation of the incidence of even symptomatic brain metastases. Brain imaging studies are not routinely performed on neurologically asymptomatic cancer patients and autopsy studies are outdated. Furthermore, while incidence rates for cancers are stable and mortality is decreasing due to earlier detection and better therapy, the incidence of brain metastases appears to be increasing. The pathophysiology of brain metastases is a complex multistage process, mediated by molecular mechanisms; from the primary organ, cancer cells must transform, grow and be transported to the CNS where they can lay dormant for various lengths of time before invading and growing further. Understanding the pathophysiology of brain metastases is of great importance, because it may lead to the development of more efficient therapies to combat brain tumor growth or to possibly make the CNS an undesirable environment for tumor progression. PMID- 16215812 TI - Symptomatic management and imaging of brain metastases. AB - Brain metastasis is the most common malignancy of the nervous system. Survival is short and the majority of patients die within 5 months after diagnosis. In this review, clinical and pathophysiological aspects of brain metastases are described, including novel radiological methods as triple-dose gadolinium enhanced MRI. Recursive partitioning analysis is a powerful tool to analyse prognosis, and recent studies contribute to subgroup division. Subsequently, treatment choices can be made, based on prognostic characteristics of the individual patient. Commonly, symptomatic therapy starts with the administration of corticosteroids, often resulting in improvement of neurological deficit. Anticonvulsants are administered in patients with symptomatic epilepsy. The risk on vascular complications in patients with brain metastases is increased and needs special attention. Treatment of psychiatric complications e.g. delirium or depression may also improve quality of life. PMID- 16215813 TI - Surgery of brain metastases--is there still a place for it? AB - The role of surgery in the treatment of metastatic brain tumors has always been a source of controversy. It was only in the early 1990s that two randomized prospective trials demonstrated that surgery plus radiation therapy improved survival in patients with single metastatic brain tumors vs. radiation therapy alone. This paper reviews these articles as well as other evidence outlining management options for multiple brain metastases. An attempt has been made to better define the role of surgery in brain metastases. The prognostic factors for brain metastases after surgery are also reviewed and the data comparing stereotactic radiosurgery to surgery is examined. A short description of surgical planning, operative techniques and tools, followed by a discussion on complication avoidance before, during, and after surgery is included. PMID- 16215814 TI - Radiotherapy and chemotherapy of brain metastases. AB - The authors have reviewed the results, the indications and the controversies regarding radiotherapy and chemotherapy of patients with newly diagnosed and recurrent brain metastases. Whole-brain radiotherapy, radiosurgery, hypofractionated stereotactic radiotherapy, brachytherapy and chemotherapy are the available options. New radiosensitizers and cytotoxic or cytostatic agents are being investigated. Adjuvant whole brain radiotherapy, either after surgery or radiosurgery, and prophylactic cranial irradiation in small-cell lung cancer are discussed, taking into account local control, survival, and risk of late neurotoxicity. Increasingly, the different treatments are tailored to the different prognostic subgroups, as defined by Radiation Therapy Oncology Group RPA Classes. PMID- 16215815 TI - Metastasis to nervous system: spinal epidural and intramedullary metastases. AB - Spinal cord epidural metastasis (SEM) is a common complication of systemic cancer with an increasing incidence. Prostate, breast and lung cancer are the most common offenders. Metastasis usually arises in the posterior aspect of vertebral body with later invasion of epidural space. Pathophysiologically, vascular insufficiency is more important than direct spinal cord compression. The most common complaint is pain, and two thirds of patients with SEM have motor signs at initial diagnosis. Currently magnetic resonance imaging is the most sensitive diagnostic tool. The optimal management of SEM is still arguable, but recent advances in surgical management of SEM and higher complication rate of surgery following radiotherapy should persuade clinicians to consider de novo surgery where possible. Radiotherapy has an important role, particularly in treatment of radiosensitive tumors and in patients who are not candidates for surgery. Novel approaches such as stereotactic radiosurgery are promising; however, response to chemotherapy depends on inherent properties of primary tumor. Recurrent SEM is a substantial problem for which surgery or repeat radiotherapy may be options. Intramedullary metastasis is rare but should be considered in patients with systemic malignancy and asymmetrical presentation of myelopathic symptoms. The prognosis is usually poor and preferred modality of treatment is radiotherapy. PMID- 16215816 TI - Dural metastases. AB - Dural metastases are found at autopsy in 8-9% of patients with advanced systemic cancer. They arise either by direct extension from skull metastases or by hematogeneous spread. Dural metastases are often clinically asymptomatic but they may produce progressive neurological deficits and sometimes subdural hematomas. MRI may be misleading when the metastasis simulates a meningioma or when a subdural hematoma masks the underlying tumor. Whenever possible, surgical removal is the most appropriate treatment. The prognosis is poor because of the progressive systemic cancer but prolonged survival has been reported in operated patients, when the systemic cancer was controlled. PMID- 16215817 TI - Skull-base metastases. AB - Metastasis to the skull-base particularly affects patients with carcinoma of the breast and prostate. Clinically, the key feature is progressive ipsilateral involvement of cranial nerves. Five syndromes have been described according to the metastatic site including the orbital, parasellar, middle-fossa, jugular foramen and occipital condyle syndromes. Magnetic resonance imaging (MRI) is nowadays the most useful examination to establish the diagnosis but plain films, CT scans with bone windows and isotope bone scans remain helpful to demonstrate bone erosion. Normal imaging studies do not exclude the diagnosis. The treatment depends on the nature of the underlying tumor. Radiotherapy is generally the standard treatment, while some patients with chemosensitive or hormonosensitive lesions benefit from chemotherapy or hormonotherapy and selected patients from surgical removal. Gamma Knife radiosurgery is sometimes a useful alternative, particularly for previously irradiated skull-base regions, and for small tumors (diameter < 30 mm). The overall prognosis is poor, with an overall median survival of about 2.5 years, probably because skull-base metastases appear late in the course of the disease. PMID- 16215818 TI - Leukemic and lymphomatous meningitis: incidence, prognosis and treatment. AB - Neoplastic meningitis (NM) is a common problem in neuro-oncology occurring in approximately 5% of all patients with cancer. Notwithstanding frequent focal signs and symptoms in NM, NM is a disease affecting the entire neuraxis and therefore staging and treatment need encompass all cerebrospinal fluid (CSF) compartments. Central nervous system (CNS) staging of NM includes contrast enhanced cranial computerized tomography (CE-CT) or magnetic resonance imaging (MR-Gd), contrast enhanced spine magnetic resonance imaging (MR-S) or computerized tomographic myelography (CT-M) and radionuclide CSF flow study (FS). Treatment of NM involves involved-field radiotherapy of bulky or symptomatic disease sites and intra-CSF drug therapy. The inclusion of concomitant systemic therapy may benefit patients with NM and may obviate the need for intra-CSF chemotherapy. At present, intra-CSF drug therapy is confined to three chemotherapeutic agents (i.e. methotrexate, cytosine arabinoside and thio-TEPA) administered by a variety of schedules either by intralumbar or intraventricular drug delivery. Although treatment of NM is palliative with an expected median patient survival of 4 to 6 months, it often affords stabilization and protection from further neurologic deterioration in patients with NM. In patients with leukemia or lymphoma, prophylaxis of the CNS is used (utilizing a combination of high-dose systemic chemotherapy and intra-CSF chemotherapy) for patients at high risk as defined by specific tumor-related laboratory markers. Using such a risk stratified approach, the late occurrence of CNS relapse has decreased dramatically attesting to the value of CNS prophylaxis. PMID- 16215821 TI - Laboratory scale bioremediation of acid mine water drainage from a disused tin mine. AB - Real acidic mine-water drainage was seeded with Acidithiobacillus ferrooxidans to catalyse the removal of iron contained therein. The addition of At. ferrooxidans increased metal precipitation kinetics and decreased the water iron content by approximately 70%. Supplementing non-sterile mine water with a bacterial growth medium accelerated metal removal by indigenous micro-organisms both at the 500 ml shake-flask and 5 l bioreactor scale. PMID- 16215822 TI - Inhibition of platelet aggregation of a mutant proinsulin chimera engineered by introduction of a native Lys-Gly-Asp-containing sequence. AB - An eight amino acid sequence, CAKGDWNC, from disintegrin barbourin, was introduced into an inactive human proinsulin molecule between the B28 and A2 sites to construct a chimeric, anti-thrombosis recombinant protein. The constructed Lys-Gly-Asp (KGD)-proinsulin gene was expressed in Escherichia coli and then purified. The KGD-proinsulin chimera protein inhibits human platelet aggregation, induced by ADP, with an IC50 value (molar concentration causing 50% inhibition of platelet aggregation) of 830 nM: and demonstrates also specific affinity to glycoprotein IIb/IIIa receptor. Its insulin receptor binding activity remains as low as 0.04% with native insulin as a control. PMID- 16215820 TI - Metastases to the peripheral nervous system. AB - Cancer metastasis can affect any part of the nervous system. When the peripheral nervous system is involved, the usual targets are cranial nerves, nerve roots and plexi. However, peripheral nerves and muscles can also be affected by compression or infiltration of neoplastic cells. This review focuses in the diagnosis and treatment of metastatic complications of cancer involving plexi, peripheral nerves and muscles. PMID- 16215823 TI - Construction of an effective protein expression system using the tpl promoter in Escherichia coli. AB - An effective protein expression system was constructed in Escherichia coli using the promoter of the tyrosine phenol-lyase (tpl) gene of Erwinia herbicola. This system involves a mutant form of the TyrR protein with an enhanced ability to activate tpl and the TutB protein with an ability to transport L-tyrosine (an inducer of Tpl). The highest expression level obtained for this system was more than twice that obtained for the tac system, although it was lower than the level obtained for the T7 system, as revealed with the lac-reporter assay and SDS polyacrylamide gel electrophoresis. PMID- 16215819 TI - Leptomeningeal metastases from solid malignancy: a review. AB - Leptomeningeal metastases (LMM) consist of diffuse involvement of the leptomeninges by infiltrating cancer cells. In solid tumors, the most frequent primary sites are lung and breast cancers, two tumors where the incidence of LMM is apparently increasing. Careful neurological examination is required to demonstrate multifocal involvement of the central nervous system (CNS), cranial nerves, and spinal roots, which constitute the clinical hallmark of the disease. Cerebro-spinal fluid (CSF) analysis is almost always abnormal but only a positive cytology or demonstration of intrathecal synthesis of tumor markers is diagnostic. T1-weighted gadolinium-enhanced sequence of the entire neuraxis (brain and spine) plays an important role in supporting the diagnosis, demonstrating the involved sites and guiding treatment. Radionuclide CSF flow studies detect CSF compartmentalization and are useful for treatment planning. Standard therapy relies mainly on focal irradiation and intrathecal or systemic chemotherapy. Studies using other therapeutic approaches such as new biological or cytotoxic compounds are ongoing. The overall prognosis remains grim and quality of life should remain the priority when deciding which treatment option to apply. However, a sub-group of patients, tentatively defined here, may benefit from an aggressive treatment. PMID- 16215824 TI - Purification and properties of an N-acetylglucosaminidase from Streptomyces cerradoensis. AB - An N-acetylglucosaminidase produced by Streptomyces cerradoensis was partially purified giving, by SDS-PAGE analysis, two main protein bands with Mr of 58.9 and 56.4 kDa. The Km and Vmax values for the enzyme using p-nitrophenyl-beta-N acetylglucosaminide as substrate were of 0.13 mM: and 1.95 U mg(-1) protein, respectively. The enzyme was optimally activity at pH 5.5 and at 50 degrees C when assayed over 10 min. Enzyme activity was strongly inhibited by Cu2+ and Hg2+ at 10 mM, and was specific to substrates containing acetamide groups such as p nitrophenyl-beta-N-acetylglucosaminide and p-nitrophenyl-beta-D-N,N' diacetylchitobiose. PMID- 16215825 TI - A modified PCR system for amplifying beta-ketoacyl-ACP synthase gene fragments with DNA from Streptomyces luteogriseus. AB - Streptomyces luteogriseus strain 099, producing a new type of macrolide antibiotic with anti-coxB6 virus and anti-HIV protease activities, was isolated from soil. PCR was optimized to amplify beta-ketoacyl-ACP synthase (KS) genes. The system was optimized around the use of higher concentrations of DMSO (15% vs. 10% v/v) and dNTP (500 microM vs. 50-200 microM) and a lower annealing temperature (55 degrees C vs. 60-70 degrees C) than the normal PCR method used to amplify high GC content DNA. PMID- 16215826 TI - Molecular cloning and tissue distribution of SF-1-related orphan receptors during sexual maturation in female goldfish. AB - The steroidogenic factor (SF)-1 gene is one of a number of orphan nuclear receptors, which is a key transcriptional regulator in vertebrate reproduction. We have isolated the SF-1 homologue cDNA from the goldfish pituitary and designed primers for SF-1 on the basis of the highly conserved regions of various known SF 1 superfamily genes. SF-1 cDNA contained 1,948 nucleotides including an open reading frame predicted to encode a protein of 503 amino acids. The distribution pattern of SF-1 in a variety of tissues during sexual maturation in female goldfish was also examined by RT-PCR. Significant variations in the relative expression of SF-1 were observed in different tissues in immature and mature female goldfish. SF-1 transcript in pituitary was significantly higher than other tissues tested in immature and mature female goldfish. Lower expression of SF-1 was observed in the liver but was not detected in brain and ovary of the immature female goldfish. Presence of SF-1 was the predominant expression in the pituitary and brain of mature female goldfish. Also, in the mature female goldfish, a weak transcript was detected in liver and ovary. Interestingly, RT-PCR analysis revealed that the expression of SF-1 became higher in the brain and weaker in the liver in maturing female goldfish. Thus, SF-1 may be regulated in goldfish brain and/or liver. Thus is also tissue-specific distribution of SF-1 during sexual maturation in female goldfish. PMID- 16215827 TI - Enhancement of isoflavone synthase activity by co-expression of P450 reductase from rice. AB - Plant cytochrome P450s interact with a flavoprotein, NADPH-cytochrome P450 reductase (CPR), to transfer electrons from NADPH. The gene for rice P450 reductase (RCPR) was cloned and expressed in Saccaromyces cerevisiae, where the specific activity of the expressed RPCR was 0.91 U/mg protein. When isoflavone synthase gene (IFS) from red clover, used as a model system of plant cytochrome P450, was co-expressed with RCPR in yeast, the production of genistein from naringein increased about 4.3-fold, indicating that the RCPR efficiently interacts with cytochrome P450 to transfer electrons from NADPH. PMID- 16215828 TI - Structural characterization of beta-glucans of Agaricus brasiliensis in different stages of fruiting body maturity and their use in nutraceutical products. AB - beta-Glucans of Agaricus brasiliensis fruiting bodies in different stages of maturity were isolated and characterized by FTIR and NMR. These fractions had greater amount of (1-->6)-beta-glucan and the (1-->3)-beta-glucan increased with fruiting bodies maturation. Yields of beta-glucans increased from 42 mg beta glucans g(-1) fruiting bodies (dry wt) in immature stage to 43 mg g(-1) in mature stage with immature spores, and decreased to 40 mg g(-1) in mature stage with spore maturation. Mature fruiting bodies, which included these glucans, have potential therapeutical benefits for use in nutraceutical products. PMID- 16215829 TI - Taxane production in suspension culture of Taxus x media var. Hicksii carried out in flasks and bioreactor. AB - Paclitaxel and 10-deacetylbaccatin III (10-DAB III) were produced in suspension cultures of Taxus x media var. Hicksii grown in shake-flasks and in a 7-l bioreactor reaching, in the bioreactor, 4.4 mg l(-1) (on day 14) and 37.5 mg l( 1) (on day 11). In shake-flasks the highest total content of paclitaxel and 10 DAB III was 7.3 mg l(-1) (on day 4) and 8.8 mg l(-1) (on day 18). Phenylalanine, at 0.05 mM, increased paclitaxel accumulation in cells cultivated in bioreactor and flasks 30-fold and 9-fold (from 0.02 mg l(-1) to 0.6 mg l(-1) and to 0.2 mg l(-1), respectively). The 10-DAB III content in cells from flasks was increased from 0.4 mg l(-1) to 1.6 mg l(-1). PMID- 16215830 TI - Regio- and stereo-selective hydroxylation of abietic acid derivatives by Mucor circinelloides and Mortierella isabellina. AB - Mucor circinelloides and Mortierella isabellina hydroxylated dehydroabietic acid (DehA). DehA was converted regio- and stereo-selectively by whole cells of Mr. circinelloides to give 2alpha-hydroxydehydroabietic acid in a 75% molar conversion yield (11 mM from 14.7 mM DehA) after 72 h in the cultivation medium containing 3% (v/v) Tween 80. With cells of Ma. isabellina, under the same conditions, 20.5 mM (6.5 g l(-1)) 2-hydroxydehydroabietic acid (alpha/beta=81/19) was formed from 26.4 mM DehA. PMID- 16215831 TI - Increased conformational and thermal stability properties for phenylalanine dehydrogenase by chemical glycosidation with end-group activated dextran. AB - A mono-aminated dextran derivative was attached to Bacillus badius phenylalanine dehydrogenase via a carbodiimide-catalyzed reaction. The optimum temperature for the conjugate was 10 degrees C higher than for native enzyme, and its thermostability was improved by 8 degrees C. The activation free energy of thermal inactivation at 45 degrees C was increased by 16.8 kJ/mol. The improved conformational stability of the modified enzyme was confirmed by fluorescence spectroscopy. PMID- 16215832 TI - Production of fungal biomass immobilized loofa sponge (FBILS)-discs for the removal of heavy metal ions and chlorinated compounds from aqueous solution. AB - A white rot basidiomycete, Phanerochaete chrysosporium, was immobilized on loofa sponge (FBILS) discs. It removed ca. 37 and 71 mg Cd (II) g(-1) from 50 and 200 mg l(-1) aqueous solutions and up to 89% of 4-chloroanisole from a 10 mg l(-1) aqueous solution. FBILS are physically strong and chemically recalcitrant, resisting temperature, mechanical agitation, and variations in pH without alteration to shape, structure or texture. PMID- 16215834 TI - Characterization of an extracellular serine protease gene from the nematophagous fungus Lecanicillium psalliotae. AB - The gene encoding a cuticle-degrading serine protease was cloned from three isolates of Lecanicillium psalliotae (syn. Verticillium psalliotae) by 3' and 5' RACE (rapid amplification of cDNA ends) method. The gene encodes for 382 amino acids and the protein shares conserved motifs with subtilisin N and peptidase S8. Comparison of translated cDNA sequences of three isolates revealed one amino acid polymorphism at position 230. The deduced protease sequence shared high degree of similarities to other cuticle-degrading proteases from other nematophagous fungi. PMID- 16215833 TI - A metal ion as a cofactor attenuates substrate inhibition in the enzymatic production of a high concentration of D-glutamate using N-acyl-D-glutamate amidohydrolase. AB - N-Acyl-D-glutamate amidohydrolase (D-AGase) was inhibited by 94 % when 1 mol/l N acetyl-DL-glutamate was used as a substrate. The addition of 1 mM Co2+ stabilized D-AGase. Moreover, the substrate inhibition was weakened to 88% with the addition of 0.4 mM Co2+ to the reaction mixture. Although D-AGase is a zinc-metalloenzyme, the addition of Zn2+ from 0.01 to 10 mM did not increase the D-glutamic acid production in the saturated substrate. Under optimal conditions, 0.38 M D glutamic acid was obtained from N-acyl-DL-glutamate with 100% of the theoretical yield after 48 h. PMID- 16215836 TI - The occurrence of mycoplasmas in the lungs of swine in Gran Canaria (Spain). AB - The study was conducted to investigate the mycoplasmal flora in the lungs of pigs with enzootic pneumonia at Gran Canaria (Spain). From 54 pneumonic lungs collected at an abattoir, 85 isolates were cultivated. On the basis of cultural and biochemical characteristics, the isolates were preliminarily identified as Mycoplasma species. Using different species-specific PCRs, 40, 27, 11 and 7 of the isolates were identified as M. hyorhinis, M. hyopneumoniae, M. hyosynoviae and M. flocculare, respectively. Nine of the M. hyopneumoniae cultures were found to be in mixed culture with M. flocculare as demonstrated by PCR. By use of a M. flocculare antiserum it was possible to eliminate M. flocculare from M. hyopneumoniae mixed cultures. This study is the first report on isolation of porcine mycoplasmas at Gran Canaria (Spain). PMID- 16215837 TI - A single-chain fragment variable recombinant antibody against F5 fimbria of enterotoxigenic Escherichia coli inhibits agglutination of horse red blood cells induced by F5 protein. AB - Bovine colibacillosis caused by enterotoxigenic Escherichia coli (ETEC) is a worldwide problem. Adhesion of ETEC to intestinal cell receptors mediated by the surface protein F5 fimbriae is the initial step in the establishment of colibacillosis. Prevention of ETEC F5(+) adhesion to enterocytes protects newborn calves against collibacillosis. On the enterocytes, the F5 fimbriae bind to a ganglioside that is also found on horse red blood cells. Thus, the presence of F5 fimbriae induces haemagglutination, which is useful as an indicator in a functional assay system. In this study, recombinant anti-F5 scFv antibody fragment produced in E. coli HB2151 reacted with F5 fimbriae in ELISA and Western immunoblot, and prevented haemagglutination induced by the binding of the F5 fimbriae to its natural host receptors on horse red blood cells. Given the ease with which recombinant antibodies can be mass-produced, the presently described scFv may hold promise as a prophylactic agent for colibacillosis. PMID- 16215839 TI - Development of a latex agglutination test using the major epitope domain of glycoprotein E of pseudorabies virus expressed in E. coli to differentiate between immune responses in pigs naturally infected or vaccinated with pseudorabies virus. AB - A 0.8 kb DNA fragment encoding the major epitope domain of glycoprotein E (gE) of pseudorabies virus (PRV) was inserted downstream of the T7 promoter of an expression vector, pET-28b, to yield the recombinant plasmid pETgE804. After induction by isopropy1-beta-D-thiogalactopyranoside (IPTG), a high level expression of fusion protein was obtained. SDS-PAGE and western immunoblotting analysis showed that the fusion protein was 38 kDa and could bind with antisera against PRV. The protein existed mainly in the form of the inclusion body. After being denatured and renatured, the protein was used to prepare the latex antigen. The concentration of antigen, temperature and time for sensitization were optimized. The latex agglutination test (LAT) was able to differentiate sera of PRV-infected pigs from those of gE-deletion vaccine-immunized pigs. The diagnostic specificity and sensitivity of the developed gE latex agglutination test (gE-LAT) were also evaluated by using sets of sera. The diagnostic specificity and diagnostic sensitivity of the gE-LAT were 96.77% and 95.76%, respectively. For comparison between gE-LAT and a commercial blocking enzyme linked immunosorbent assays (ELISA), 260 serum samples were tested. The coincidence frequency of both assays was 96.94% (252/260). No significant difference was found between the two methods (p>0.05). For comparison between the abilities of gE-LAT and gE-ELISA to detect sera with low titres of gE-specific antibody, 66 sera from 22 pigs were tested. The data indicate that the gE-LAT is of similar sensitivity to gE-ELISA. These results indicate that gE-LAT using recombinant gE might be very useful as a routine screening method for the differential diagnosis of PRV infection. PMID- 16215838 TI - Comparison of expression of glucokinase gene and activities of enzymes related to glucose metabolism in livers between dog and cat. AB - Plasma glucose and immunoreactive insulin (IRI) concentrations and activities of enzymes related to glucose metabolism in livers were measured in dogs and cats. Nucleotide sequences of the conserved region of glucokinase (GK) cDNA that contained ATP- and glucose-binding domains were determined in canine liver and feline pancreas for design of the species-specific oligonucleotide primers for reverse transcription-polymerase chain reaction (RT-PCR) analysis. There were no significant differences in plasma glucose and IRI concentrations between dogs and cats. In feline liver, although GK activities were not detected, activities of hexokinase, fructokinase, pyruvate kinase, glucose-6-phosphate dehydrogenase, fructose-1,6-bisphosphatase and glucose-6-phosphatase were significantly higher than those in canine liver. The partial sequences of canine liver GK and feline pancreas GK cDNA were respectively 88% and 89% identical with the rat liver GK cDNA. Expression of GK gene was observed in canine liver and pancreas and feline pancreas with RT-PCR using species specific primers based on the cDNA sequences. PMID- 16215841 TI - Aflatoxin B1 binding to sorbents in bovine ruminal fluid. AB - A recent approach to the problem of contamination of agricultural products by aflatoxin B(1) (AFB(1)) is to add non-nutritional adsorbents to animal diets in order to sequester ingested aflatoxins. We conducted in vitro experiments to develop a rapid and cheap model using ruminal fluid to assess the ability of sorbent materials to bind AFB(1). Seven sorbents (hydrated sodium calcium aluminosilicate; clinoptilolite; zeolite; two types of bentonite; sepiolite; and PHIL 75), commonly added to bovine diets were incubated in water and ruminal fluid in the presence of AFB(1). Hydrated sodium calcium aluminosilicate, sepiolite and one of the bentonites bound 100% of the AFB(1) in the presence of both ruminal fluid and water; clinoptilolite bound about 80% of AFB(1) in both liquids; whereas the affinities for the mycotoxin of zeolite (50%) and the other sample of bentonite (60%) in water seem to be increased by about 40% in ruminal fluid incubations. PHIL 75 had the poorest binding ability: about 30% in water and 45% in ruminal fluid. In view of the differences in toxin binding in water and ruminal fluid, it is preferable to use the ruminal fluid model for the in vitro pre-screening of sorbent materials potentially useful as adjuvants to ruminant feeds. PMID- 16215840 TI - Canine epidermal langerhans cells express alpha and gamma but not beta chains of high-affinity IgE receptor. AB - Epidermal Langerhans cells (LC) express a high-affinity receptor for IgE (FcepsilonRI), consisting of two chains (alpha and gamma chains) in humans that allows LC to perform Fc receptor-mediated uptake of allergens. We found that canine LC express alpha and gamma chains but not beta chain of FcepsilonRI, identical to human but not to mouse LC, which do not express functional FcepsilonRI (only gamma chain is expressed). This finding indicates that canine LC have FcepsilonRI-mediated function similar to or identical to human LC, raising the possibility that canine species provides a better model than mouse to understand the pathogenesis of human atopic dermatitis and investigate the therapeutic effect of drugs. PMID- 16215842 TI - Capsid protein genetic analysis and viral spread to the spinal cord in cats experimentally infected with feline calicivirus (FCV). AB - We investigated primitively the molecular basis of the neural spread of a feline calcivirus isolate (FCV-S) from the spinal cord of a cat that died after manifesting excitation. Experimental infections of cats with three clones from parent virus isolate FCV-S, isolated based on plaque size, were performed, and virus recovery from the spinal cord and the nucleotide and predicted amino acid sequences of the viral capsid protein region (ORF2) were compared. In the experimental infection with the one-time cloned virus (C1L1) isolated from a large plaque, the C1L1 was recovered from the spinal cord. In contrast, seven times cloned C6L7 (from large plaque) and five-times cloned C5S2 (isolated from small plaque) were not recovered from the spinal cord. Genetic analysis of the capsid protein gene of the three viral clones revealed that four bases were different and two amino acids were different at positions 34 (Val in C6L7 and Ala in C1L1 and C5S2) and 46 (Leu in C6L7 and Pro in C1L1 and C5S2) between C6L7 (with large plaque) and C5S2 (with small plaque). The amino acid at position 434 of C1L1 was different from those of C6L7 and C5S2 (Gly in C1L1, D (Asp) in C6L7 and C5S2). From these results, the plaque size seemed not to be related to the spread of virus to the spinal cord. Clone C1L1, which spread to the spinal cord, had a difference of one amino acid from the other two clones, which may be related to the ability to spread to the spinal cord. PMID- 16215843 TI - Ribotyping of Indian isolates of Pasteurella multocida based on 16S and 23S rRNA genes. AB - The applicability of ribotyping based on 16S and 23S rRNA was evaluated for molecular epidemiological studies. Forty-eight isolates of Pasteurella multocida isolated from different hosts and geographical locations and one reference isolate were ribotyped. Only four ribotypes were found. All the isolates including reference isolate from wild carnivores had the same ribotype, though they had different serotypes. The isolate from a tiger had one band in addition to the bands present in the major ribotype. The isolates from lions represented two ribotypes; of these ribotypes, one (r2) had an additional band of 3.6 kbp, which was absent in all other ribotypes. The second ribotype (r4) from a lion had one band missing (6 kbp) that was present in the other ribotypes. These isolates were further typed using ERIC-PCR and REP-PCR. With ERIC-PCR and REP-PCR, higher D values of 0.83 and 0.89 were obtained. The current study revealed that ribotyping is not a very efficient typing tool for use in molecular epidemiology for differentiation of isolates. PMID- 16215844 TI - Cloning and characterization of tbpA gene encoding transferrin-binding protein (TbpA) from Pasteurella multocida serogroup B:2 (strain P52). PMID- 16215845 TI - Using stochastic differential equations for PK/PD model development. AB - A method for PK/PD model development based on stochastic differential equation models is proposed. The new method has a number of advantages compared to conventional methods. In particular, the new method avoids the exhaustive trial and-error based search often conducted to determine the most appropriate model structure, because it allows information about the appropriate model structure to be extracted directly from data. This is accomplished through quantification of the uncertainty of the individual parts of an initial model, by means of which tools for performing model diagnostics can be constructed and guidelines for model improvement provided. Furthermore, the new method allows time-variations in key parameters to be tracked and visualized graphically, which allows important functional relationships to be revealed. Using simulated data, the performance of the new method is demonstrated by means of two examples. The first example shows how, starting from a simple assumption of linear PK, the method can be used to determine the correct nonlinear model for describing the PK of a drug following an oral dose. The second example shows how, starting from a simple assumption of no drug effect, the method can be used to determine the correct model for the nonlinear effect of a drug with known PK in an indirect response model. PMID- 16215846 TI - Effect of P-selectin inhibition on leukocyte-endothelium interaction and survival after global cerebral ischemia. AB - Cerebral ischemia induces activation of leukocyte-endothelium interactions requiring upregulation of specific adhesion molecules including the selectins. The aim of the current study was to elucidate the therapeutic potency of P selectin blockade on microcirculatory disturbances and secondary brain damage after global cerebral ischemia. Global cerebral ischemia for 15 minutes was induced in Mongolian gerbils. Functional blockade of P-selectin was achieved by pretreatment with the antibody RB 40.34 (2 mg/kg, n = 7). In vivo observation of brain microcirculation was performed by epifluorescence microscopy of a cranial window. Survival was assessed daily up to 4 days after ischemia. In the control group leukocyte rolling increased during reperfusion with a maximum at 3 h (28 +/ 14 x 100 microm(-1) x min(-1)) and was significantly reduced by the P-selectin antibody (13 +/- 9 x 100 microm(-1) x min(-1), p < 0.05). No effect on firm leukocyte adhesion was observed (4 +/- 3 vs. 2 +/- 1 x 100 microm(-1) x min(-1)). The survival of animals that received the Pselectin antibody (28 %) was significantly reduced compared with controls (71 %). Anti-P-selectin antibody reduces leukocyte rolling but has no positive effect on survival. Our data question the role of the inflammatory response in the development of secondary brain damage and do not support this kind of therapeutical approach in global cerebral ischemia. PMID- 16215847 TI - Human antimicrobial peptides: defensins, cathelicidins and histatins. AB - Antimicrobial peptides, which have been isolated from many bacteria, fungi, plants, invertebrates and vertebrates, are an important component of the natural defenses of most living organisms. The isolated peptides are very heterogeneous in length, sequence and structure, but most of them are small, cationic and amphipathic. These peptides exhibit broad-spectrum activity against Gram-positive and Gram-negative bacteria, yeasts, fungi and enveloped viruses. A wide variety of human proteins and peptides also have antimicrobial activity and play important roles in innate immunity. In this review we discuss three important groups of human antimicrobial peptides. The defensins are cationic non glycosylated peptides containing six cysteine residues that form three intramolecular disulfide bridges, resulting in a triple-stranded beta-sheet structure. In humans, two classes of defensins can be found: alpha-defensins and beta-defensins. The defensin-related HE2 isoforms will also be discussed. The second group is the family of histatins, which are small, cationic, histidine rich peptides present in human saliva. Histatins adopt a random coil conformation in aqueous solvents and form alpha-helices in non-aqueous solvents. The third group comprises only one antimicrobial peptide, the cathelicidin LL-37. This peptide is derived proteolytically from the C-terminal end of the human CAP18 protein. Just like the histatins, it adopts a largely random coil conformation in a hydrophilic environment, and forms an alpha-helical structure in a hydrophobic environment. PMID- 16215848 TI - Mutation detection in DNA oligonucleotides based on a Guanine quenching method coupled with enzymatic digestion of single-stranded DNA. AB - Fluorescence quenching by guanine allows DNA hybridization to be monitored and any point mutations in oligonucleotides to be detected. However, fluorescence quenching is often affected by untargeted guanine located in a protruding end (single-strand DNA) of the probe-target DNA duplex resulting in an unsatisfactory sensitivity. In the present study, we used enzymatic digestion of the protruding end of a probe-target DNA duplex to avoid interference by untargeted guanine on fluorescence quenching for detection of a nucleobase mutation. Enzymatic digestion of the protruding end of the DNA duplex fully prevented interference by untargeted guanine, and produced a marked difference in the quenching ratios (36% for wild-type, and 0% for mutant). PMID- 16215849 TI - Cloning and characterization of OsORC2, a new member of rice origin recognition complex. AB - In eukaryotic cells, the origin recognition complex (ORC) governs the initiation site of DNA replication and formation of the prereplication complex. The isolation, characterization and tissue-specific expression of a putative ORC subunit 2 (OsORC2) in Oryza sativa is described here. A novel cDNA fragment encoding rice ORC2 was isolated by screening the subtractive library, which had a higher expression level in inflorescence meristem than in shoot apical meristem. The full-length cDNA of rice ORC2 was obtained by the method of rapid amplification of cDNA ends, which contained an 1140 bp open reading frame encoding a 379 amino acid polypeptide. Sequence alignment shows that there is a high homology between the deduced amino sequence of OsORC2 and maize ORC2 (85%). The tissue-specific expression pattern of OsORC2 reveals that it is abundant in roots, seedling and inflorescence meristem, while its expression level is much lower in mature leaves and shoot. PMID- 16215850 TI - Timing of bifidobacterium administration influences the development of allergy to ovalbumin in mice. AB - C3H/HeJ mice were sensitized with ovalbumin (OVA) and choleratoxin (CT) for 5 weeks, and then Bifidobacterium bifidum BGN4 was administered continuously for 7 weeks, starting 2 weeks before (pre-treatment group) and 2 weeks after (post treatment group) the initial sensitization. After sensitization, the OVA-induced (sham group) mice showed growth inhibition and had scab-covered tails which was associated with serum levels of 9887+/-175 ng OVA-specific IgE/ml and 758+/-525 ng IgG1/ml. The sera of the pre-treatment group had 4805+/-245 ng OVA-specific IgE/ml and 193+/-87 ng IgG1/ml, as well as less severe tail symptoms. The sera of the post-treatment group had 5723+/-207 ng OVA-specific IgE/ml but the IgG1 and IgG2a levels were the same as those of the sham group. In spleen cultures, both pre-treatment and post-treatment increased the levels of IFN-gamma but decreased the levels of IL-6 and IL-18. Taken together, the in vivo and in vitro results show that treatment with Bifidobacterium before OVA sensitization suppresses or modulates the allergic response more effectively than treatment with Bifidobacterium following OVA sensitization. PMID- 16215851 TI - Expression, purification and characterization of a recombinant beta-glucosidase from Volvariella Volvacea. AB - For the first time, a beta-glucosidase gene from the edible straw mushroom, Volvariella volvacea V1-1, has been over-expressed in E. coli. The gene product was purified by chromatography showing a single band on SDS-PAGE. The recombinant enzyme had a molecular mass of 380 kDa with subunits of 97 kDa. The maximum activity was at pH 6.4 and 50 degrees C over a 5 min assay. The purified enzyme was stable from pH 5.6-8.0, had a half life of 1 h at 45 degrees C. The beta glucosidase had a K(m) of 0.2 mM: for p-nitrophenyl-beta-D: -glucopyranoside. PMID- 16215852 TI - Metabolic flux analysis of diterpene biosynthesis pathway in rice. AB - Relative transcript levels of eight rice diterpene cyclases at the branch points of gibberellins and phytoalexins biosynthesis pathway were measured by reverse transcription quantitative PCR. Metabolic flux analysis by the distribution ratio of common substrate showed that UV-irradiation of etiolated rice seedlings decreased the flux for primary metabolism of gibberellins biosynthesis by half (from 62 to 27%) and 41% of geranylgeranyl pyrophosphate was used for induction of pimaradiene intermediate as the major phytoalexin. In comparison, light illumination used almost all geranylgeranyl pyrophosphate (96%) for gibberellin biosynthesis to stimulate the plant growth and strongly repressed the metabolic flux for phytoalexins biosynthesis. PMID- 16215853 TI - Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from gluconate and glucose by recombinant Aeromonas hydrophila and Pseudomonas putida. AB - Aeromonas hydrophila 4AK4 and Pseudomonas putida GPp104 were genetically engineered to synthesize poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) using gluconate and glucose rather than fatty acids. A truncated tesA gene, encoding cytosolic thioesterase I of Escherichia coli which catalyzes the conversion of acyl-ACP into free fatty acids, was introduced into A. hydrophila 4AK4. When grown in gluconate, the recombinant A. hydrophila 4AK4 synthesized 10% (w/w) PHBHHx containing 14% (mol/mol) 3-hydroxyhexanoate. If additional PHBHHx synthesis genes, phaPCJ, were over-expressed with the truncated tesA in A. hydrophila 4AK4, the PHBHHx content increased to 15% (w/w) and contained 19% (mol/mol) 3-hydroxyhexanoate. Recombinant P. putida GPp104 harboring phaC encoding PHBHHx synthase of A. hydrophila, phaB encoding acetoacetyl-CoA reductase of Wautersia eutropha and phaG encoding 3-hydroxyacyl-ACP-CoA transferase of P. putida, synthesized 19% (w/w) PHBHHx containing 5% (mol/mol) 3 hydroxyhexanoate from glucose. The results suggest that the engineered pathways were applicable to synthesize PHBHHx from unrelated carbon sources such as gluconate and glucose. PMID- 16215854 TI - Hydroxynitrile lyase catalysis in ionic liquid-containing systems. AB - The cleavage of mandelonitrile catalysed by hydroxynitrile lyases (HNL) from Prunus amygdalus (PaHNL) and Manihot esculenta (MeHNL) proceeded more rapidly in monophasic aqueous media containing 1-propyl-3-methylimidazolium tetrafluoroborate [C4MIm][BF4] than in media containing acetonitrile or THF. Both HNLs were much more thermostable in [C4MIm][BF4] than in acetonitrile or THF. The addition of each of the four ionic liquids 1-butyl-, 1-pentyl- and 1-hexyl-3 methylimidazolium tetrafluoroborates at 2-6% (v/v in the aqueous phase) increased both the enzyme activity and the product e.e. in the PaHNL-catalysed transcyanation in an aqueous/DIPE biphasic system. However, MeHNL was inactivated by the ionic liquids, as indicated by the decreased reaction rate, substrate conversion and product e.e. PMID- 16215855 TI - Differential patterns of constitutive intracellular laccases of the vegetative phase of Pleurotus species. AB - The constitutive intracellular laccase activity of ten strains of Pleurotus spp. was determined in vitro and by zymograms, using different substrates. Differences in the in vitro activities were observed between all the strains; however, zymogram patterns were only similar for strains within same species, independently of any of the three substrate (2,6-dimethoxyphenol, p-anisidine or o-tolidine) used. The differences observed in the number and positions of the isoforms in the gel suggest that laccase zymograms can be used to differentiate species of this organism. PMID- 16215857 TI - Repeated batch cultivation of Ralstonia eutropha for Poly (beta-hydroxybutyrate) production. AB - Batch cultivation of Ralstonia eutropha NRRL B14690 attained 21 g biomass l(-1) and 9.4 g poly(beta-hydroxybutyrate) l(-1) (0.45 g PHB g(-1 )dry wt(-1)) in 60 h. Repeated batch operation (empty-and-fill protocol) to remove 20% (v/v) of the culture broth and to supplement an equal volume of fresh media resulted in 49 g biomass l(-1) and 25 g PHB l(-1) (0.51 g PHB g(-1) dry wt(-1)) with an overall productivity of 0.42 g PHB l(-1 )h(-1 )in 67 h. In the two cycles of repeated batch fermentation there was a 3-fold increase in productivity as compared to batch. PMID- 16215856 TI - Fed-batch cultivation of Mucor indicus in dilute-acid lignocellulosic hydrolyzate for ethanol production. AB - Mucor indicus fermented dilute-acid lignocellulosic hydrolyzates to ethanol in fed-batch cultivation with complete hexose utilization and partial uptake of xylose. The fungus was tolerant to the inhibitors present in the hydrolyzates. It grew in media containing furfural (1 g/l), hydroxymethylfurfural (1 g/l), vanillin (1 g/l), or acetic acid (7 g/l), but did not germinate directly in the hydrolyzate. However, with fed-batch methodology, after initial growth of M. indicus in 500 ml enzymatic wheat hydrolyzate, lignocellulosic hydrolyzate was fermented with feeding rates 55 and 100 ml/h. The fungus consumed more than 46% of the initial xylose, while less than half of this xylose was excreted in the form of xylitol. The ethanol yield was 0.43 g/g total consumed sugar, and reached the maximum concentration of 19.6 g ethanol/l at the end of feeding phase. Filamentous growth, which is regarded as the main obstacle to large-scale cultivation of M. indicus, was avoided in the fed-batch experiments. PMID- 16215858 TI - Biosynthesis and characterization of poly(3-hydroxybutyrate-co-3- hydroxyhexanoate) from palm oil products in a Wautersia eutropha mutant. AB - Palm kernel oil, palm olein, crude palm oil and palm acid oil were used for the synthesis of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] by a mutant strain of Wautersia eutropha (formerly Ralstonia eutropha) harboring the Aeromonas caviae polyhydroxyalkanoate (PHA) synthase gene. Palm kernel oil was an excellent carbon source for the production of cell biomass and P(3HB-co-3HHx). About 87% (w/w) of the cell dry weight as P(3HB-co-3HHx) was obtained using 5 g palm kernel oil/l. Gravimetric and microscopic analyses further confirmed the high PHA content in the recombinant cells. The molar fraction of 3HHx remained constant at 5 mol % regardless of the type and concentration of palm oil products used. The small amount of 3HHx units was confirmed by 13C NMR analysis. The number average molecular weight (M(n)) of the PHA copolymer produced from the various palm oil products ranged from 27 0000 to 46 0000 Da. The polydispersity was in the range of 2.6-3.9. PMID- 16215859 TI - Synthesis of guaiacol-alpha-D: -glucoside and curcumin-bis-alpha-D: -glucoside by an amyloglucosidase from Rhizopus. AB - Guaiacol (2-methoxyphenol) and curcumin [1E,6E-1,7-di(4-hydroxy-3-methoxy-phenyl) 1,6-heptadiene-3,5-dione] were converted into their corresponding glucosides using glucose and an amyloglucosidase from Rhizopus. Guaiacol-alpha-D: -glucoside yields ranged from 3 to 52% with the highest at pH 7.0. Curcumin-bis-alpha-D: glucoside yields ranged from 3 to 48% with the highest at pH 4.0 with 50% (w/w D: -glucose) of enzyme. The phenolic hydroxyl group of guaiacol and both phenolic hydroxyl groups of curcumin were glucosylated at the C1 carbon of alpha-D: glucose indicating that the enzymatic reaction is stereospecific. Both guaiacol alpha-D: -glucoside and curcumin-bis-alpha-D: -glucosides had antioxidant activities. PMID- 16215860 TI - Optimal light regime for the cultivation of transgenic Laminaria japonica gametophytes in a bubble-column bioreactor. AB - Fluctuating light intensity had a more significant impact on growth of gametophytes of transgenic Laminaria japonica in a 2500 ml bubble-column bioreactor than constant light intensity. A fluctuating light intensity between 10 and 110 microE m(-2) s(-1), with a photoperiod of 14 h:10 h light:dark, was the best regime for growth giving 1430 mg biomass l(-1). PMID- 16215861 TI - Use of a novel immobilization yeast system for winemaking. AB - Penicillium was used to immobilize Saccharomyces cerevisiae, without using physico-chemical external supports, to form yeast biocapsules. The biocapsules, once the Penicillium was killed by the ethanol produced, were used in a grape must fermentation. Must fermentation was carried out for 160 h with the biocapsules and for 300 h with free yeast cells. Acetaldehyde (84 vs. 63 mg/l), isobutanol (217 vs. 194 mg/l), L: -proline (7.7 vs. 6.5 mM: ) and aspartic acid (0.42 vs. 0 mM: ) in final wine were higher with the biocapsules than with free cells. PMID- 16215863 TI - Intake of fruits and vegetables, carotenoids, folate, and vitamins A, C, E and risk of bladder cancer among women (United States). AB - OBJECTIVE: To examine the relation between fruits and vegetables, carotenoids, folate, and vitamins A, C, E and the risk of bladder cancer in a prospective study of women. METHODS: A total of 237 incident bladder cancer cases were documented during 20 years of follow-up among 88,796 women enrolled in the Nurses' Health Study. Dietary intake was assessed by food-frequency questionnaires every two to four years and incident diagnosis of bladder cancer was ascertained every two years. Cox proportional hazard models were used to estimate incidence rate ratios (RR) and 95% confidence intervals (CI) for bladder cancer risk, adjusting for age, pack-years of smoking, current smoking, and total caloric intake. RESULTS: Consumption of total fruits and vegetables was not associated with bladder cancer risk (RR = 1.08, 95% CI = 0.70-1.65, for > 5.5 compared to < 2.5 servings per day). Similarly, dietary intakes of carotenoids, folate, and vitamins A, C, E, were not related to bladder cancer risk. No association was observed between supplemental intake of multivitamins, vitamins A, C, E and bladder cancer risk. CONCLUSIONS: We did not observe any association for fruit and vegetable consumption or vitamin intake and bladder cancer risk among women. PMID- 16215864 TI - Genetic, anthropometric, and lifestyle factors associated with IGF-1 and IGFBP-3 levels in Hispanic and non-Hispanic white women. AB - Circulating concentrations of IGF-I and IGFBP-3 are associated with risk of pre menopausal breast cancer. Racial differences in levels of these factors have been reported, and determinants of IGF-I and IGFBP-3 levels within racial and ethnic groups are unclear. In this study we examine genetic, anthropometric, diet, and lifestyle factors that may predict serum levels of IGF-I and IGFBP-3 among Hispanic and non-Hispanic white women. A sample of healthy controls participating in the SHINE (Southwest Hormone, Insulin, Nutrition, and Exercise Study) case control breast cancer in Arizona, Colorado, New Mexico, and Utah were included in these analyses. Subjects included 210 Hispanic and 284 non-Hispanic white women. Hispanic women had significantly lower levels of IGFBP-3 (mean=3764.3 mcg/ml) after adjusting for age, body size, physical activity, menopausal status, and dietary factors than non-Hispanic white women (mean = 4058.0 mcg/ml; p<0.01). The CC genotype of the -202 A>C polymorphism of the IGFBP3 gene was associated with lower IGFBP-3 levels in both ethnic groups. The frequency of the IGFBP3 C allele differed between Hispanic (0.65) and non-Hispanic white women (0.53), but serum levels of IGFBP-3 were lower for Hispanic women than non-Hispanic after accounting for IGFBP3 genotype. Body size indicators, vigorous physical activity, and dietary factors appeared to influence serum levels of IGF-1 and the ratio of IGF-1 to IGFBP-3 in pre-menopausal women more than in post-menopausal women. On the other hand, using aspirin/NSAIDs appeared to increase IGFBP-3 levels significantly among pre-menopausal Hispanic women. Results from this study suggest that differences in IGFBP-3 levels exist in Hispanic and non-Hispanic white women. These differences could be due to the combined effects of genetic and behavioral factors which could account for ethnic differences in the risk of breast cancer and other chronic diseases. PMID- 16215865 TI - Reducing cigarette smoking among working adolescents: results from the SMART study. AB - OBJECTIVE: The SMART Teens Against the Risks of Tobacco Study was designed to test the feasibility and efficacy of tobacco control intervention methods for employed teens. METHODS: A randomized controlled pilot study tested the efficacy of a behavioral intervention delivered between September, 1999, and August, 2000. Baseline and final survey data were collected on 560 teens in four intervention and five control stores. RESULTS: Although smoking prevalence decreased and intention to quit increased more among teens in the intervention stores compared to those in the control stores, the differences were not statistically significant. CONCLUSIONS: The worksite holds promise as a possible venue for tobacco prevention and cessation interventions for youth although further research is needed to increase the efficacy of interventions for this setting. PMID- 16215866 TI - Glycemic load, glycemic index, and the risk of breast cancer among Mexican women. AB - OBJECTIVE: The amount and composition of dietary carbohydrates is a major determinant of postprandial blood glucose and insulin, and risk of breast cancer has been positively associated with plasma levels of insulin and insulin-like growth factor 1. We sought to evaluate dietary glycemic load (GL) and overall glycemic index (GI) in relation to breast cancer risk in Mexican women. METHODS: We examined dietary GL and overall GI and breast cancer risk among 475 women with histologically-confirmed breast cancer and a random sample of 1391 women from Mexico City households. Diet was assessed using a food frequency questionnaire adapted to the Mexican population. RESULTS: The multivariate adjusted or for all women comparing the highest quartile of dietary GL with the lowest quartile was 1.62 (95% CI 1.13-2.32; p-test for trend = 0.02) with a significant trend. In postmenopausal women, the multivariate adjusted or comparing the extreme quartiles was 2.18 (95% CI 1.34-3.55; p-test for trend=0.005). Overall GI was not significantly associated with risk of breast cancer. CONCLUSION: High intake of rapidly absorbed carbohydrate appears to play an important role in the risk of breast cancer in Mexican women. PMID- 16215867 TI - Patient perspectives on research recruitment through cancer registries. AB - OBJECTIVE: To gain preliminary insight into patients' levels of awareness and preferences about research recruitment through cancer registries. METHODS: We developed four questions about the North Carolina Central Cancer Registry's educational brochure and about patient preferences regarding physician involvement in research recruitment. These questions were included in the baseline interview conducted among 100 consecutively enrolled participants in the North Carolina Colorectal Cancer Care Outcomes Research Study, an ongoing observational study. RESULTS: Patients who read the Registry's educational brochure generally reported that it helped them understand that a researcher could contact them, but only about one-fourth of patients recalled receiving and reading the brochure. Over two-thirds of patients said they preferred that researchers contact them directly about their interest in research participation, rather than checking with their physician first. Among patients who wanted their physician involved, most preferred a physician notification rather than a physician permission approach. CONCLUSIONS: Registry policies about patient education and physician involvement can have an important impact on researchers' ability to conduct population-based studies. Understanding patient perspectives is key to developing balanced policies that protect patients' privacy, as well as facilitate their opportunities to make autonomous decisions about participating in research. PMID- 16215868 TI - Colorectal cancer incidence among female textile workers in Shanghai, China: a case-cohort analysis of occupational exposures. AB - Previous studies have suggested increased risks of colorectal cancers among textile industry workers, potentially related to synthetic fibers. To investigate risks of colon and rectum cancers in relation to these and other textile industry exposures, we conducted a case-cohort study nested within a cohort study of female employees from the Shanghai Textile Industry Bureau (STIB). Cox proportional hazard regression modeling was used to estimate hazard ratios (HR) for colon and rectum cancers associated with duration of employment (e.g., 0, >0 to <10, 10 to <20 years, > or =20 years) in various jobs classified according to process type and exposures to specific agents. Our findings indicate that certain long term exposures may pose increased risk of colorectal cancers, especially dyes and dye intermediates with colon cancer (> or =20 years exposure versus never, HR=3.9; 95% CI: 1.4-10.6), and maintenance occupation (HR = 2.3; 95% CI: 1.0-5.7) and metals exposure (HR = 2.0; 95% CI: 1.1-3.6) with rectum cancer. A decreased risk of rectum cancer was associated with exposure to natural fibers such as cotton (HR = 0.7; 95% CI: 0.5-0.9), and a trend of decreasing rectum cancer incidence was observed by category of cumulative quantitative cotton dust or endotoxin exposures, when exposures were lagged by 20 years. PMID- 16215869 TI - Association of vasectomy and prostate cancer among men in a Maryland cohort. AB - OBJECTIVES: To evaluate the association of vasectomy with prostate cancer. METHODS: Participants were male members of the CLUE II cohort followed since 1989. On a questionnaire mailed in 1996, the men were asked if they had had a vasectomy and their age at vasectomy. Between 1996 and April 2004, 78 prostate cancer cases were confirmed among the 3373 men who were at least 35 years old at baseline and who completed the questions about vasectomy. Cox proportional hazards regression was used to estimate age-adjusted hazard ratios (HR) of prostate cancer. RESULTS: The HR for prostate cancer for men who had had a vasectomy was 2.03 (95% CI: 1.24-3.32). Risk of low-grade disease (HR=2.87; 95% CI 1.49-5.54), but not high-grade disease (HR=0.99; 95% CI 0.36-2.76), was higher in men who had had a vasectomy. No statistically significant associations were observed for low- or high-stage disease. The association for vasectomy was more pronounced in men who were 40 years at the time of vasectomy (HR=2.63; 95% CI 1.40-4.94) than in men who were younger at vasectomy. CONCLUSIONS: The results from this prospective study suggest a positive association between vasectomy and prostate cancer, especially low-grade disease. PMID- 16215870 TI - The estrogen receptor alpha gene and breast cancer risk (The Netherlands). AB - OBJECTIVE: In this study we aimed to investigate whether the PvuII, XbaI and B variant polymorphisms in the estrogen receptor alpha gene (ER-alpha) are associated with an increased risk of breast cancer in postmenopausal women, and whether the effect of high estradiol (E2) levels on breast cancer risk is altered by these polymorphisms. The selection of these polymorphisms was based on previously published associations with osteoporosis and spontaneous abortions. METHODS: The effect of the three polymorphisms on breast cancer risk was studied using a case-cohort design nested within a large population-based cohort study (n = 9349) in the Netherlands (the DOM-cohort). In total 380 incident breast cancer cases and a subcohort of 422 women were genotyped by RFLP or ASO hybridization methods. RESULTS: Women with the PvuII pp genotype had a 1.5 times non significant increased risk of breast cancer (95% CI: 0.94-2.42; p(trend) = 0.09) compared to women with the PP genotype. The Pp or pp genotype in combination with high E2 levels raised breast cancer risk significantly when compared to women with low E2 levels and the PP genotype (RR=2.26; 95% CI: 1.24-4.13). This interaction was statistically significant on the multiplicative scale (p = 0.01). The XbaI genotype (RR(xx versus XX) = 1.19; 95% CI: 0.73-1.95) and the B' allele (RR(BB'+B'B' versus BB) = 0.87; 95% CI: 0.56-1.33) were not associated with breast cancer risk. CONCLUSION: The results of this study suggest that the PvuII polymorphism in the ER-alpha, or another mutation in linkage disequilibrium with PvuII, in combination with high E2 levels increases breast cancer risk in postmenopausal women. PMID- 16215871 TI - Anthropometrics, physical activity, related medical conditions, and the risk of non-hodgkin lymphoma. AB - BACKGROUND: Recent reports suggest that obesity, or conditions associated with obesity, might be risk factors for non-Hodgkin lymphoma (NHL), a cancer with dramatically increasing incidence in western countries over the last several decades. Physical inactivity increases the risk of obesity and of type 2 diabetes, but there are few data on the association of physical activity with risk of NHL. METHODS: We evaluated these factors in a population-based case control study conducted in Detroit, Iowa, Los Angeles, and Seattle from 1998 to 2000. Incident HIV-negative NHL cases aged 20-74 years were rapidly reported in each area (n = 1321). Controls were identified through random digit dialing and Medicare files, and were frequency matched to cases on sex, age, race, and study site (n = 1057). Risk factor data were collected by in-person interviews and self administered questionnaires. Unconditional logistic regression was used to estimate the odds ratio (OR) and 95% confidence intervals (CI), adjusted for age, sex, race and study center. RESULTS: High body mass index (OR = 1.73 for 35+ versus <25 kg/m2; 95% CI 1.15-2.59) and history of gallstones (OR = 1.95, 95% CI 1.11-3.40) were positively associated with diffuse NHL, but were not associated with follicular or all NHL combined. Height was positively associated with risk of all NHL combined (OR = 1.38 for >70 versus <65 inches; 95% CI 0.98-1.94), and positive associations were apparent for both diffuse and follicular NHL. Non occupational physical activity was inversely associated with risk of all NHL combined (ORs with increasing level: 1, 0.75, 0.71, 0.55, 0.68; p-trend = 0.04) and for diffuse and follicular NHL. We observed no association of total energy intake, type 2 diabetes, or hypertension with risk of NHL. In a multivariable model to predict risk of diffuse NHL, BMI (OR = 2.15 for 35+ versus <25 kg/m2; 95% CI 1.09-4.25) and height (OR = 1.63 for 71+ versus <65 inches; 95% CI 0.75 3.57) were positively associated with risk while physical activity was weakly and inversely associated risk (ORs with increasing level: 1, 0.76, 0.72, 0.78, 0.82; p-trend = 0.9). CONCLUSION: BMI and history of gallstones were positively associated with risk of diffuse NHL, supporting a role for obesity in this NHL subtype. Height was positively associated with NHL risk across subtypes, and suggests a role for early life nutrition in NHL risk. Non-occupational physical activity was only weakly and inversely associated with NHL risk after adjustment for obesity, height and alcohol use. PMID- 16215872 TI - Personal and occupational exposure to organic solvents and risk of non-Hodgkin's lymphoma (NHL) in women (United States). AB - OBJECTIVES: The authors assessed whether home and occupational exposure to organic solvents is associated with risk of NHL in women. METHODS: A population based, incidence case-control study was conducted in upstate New York, involving 376 NHL cases and 463 population controls selected from the Medicare beneficiary files and State driver's license records. Exposure information was obtained by telephone interview. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using an unconditional logistic regression model, adjusting for a number of risk factors for NHL. RESULTS: Overall, history of exposure to organic solvents was not associated with the risk of NHL. A statistically significant increase in risk associated with occupational exposure was observed only for the subjects whose first exposure occurred before 1970 (OR=1.87, 95% CI 1.03-3.40). When occupational and home exposures to paint thinners/turpentine were combined and analyzed together, the risk of NHL associated with any exposure, compared to no exposure at either job or home, was a statistically significantly increased (OR=1.46, 95% CI: 1.05-2.03). This observation was more pronounced for B-cell lymphoma and for low-grade lymphoma with ORs of 1.52 (95 CI: 1.08-2.14) and 2.20 (95% CI; 1.42-3.41), respectively. CONCLUSIONS: The results of this case-control study do support of a major role of organic solvents in the development of NHL among women currently living in the US. However, relatively intensive exposure in past occupations and use of paint thinners/turpentine may deserve further investigation. PMID- 16215873 TI - MnSOD Val-9Ala genotype, pro- and anti-oxidant environmental modifiers, and breast cancer among women on Long Island, New York. AB - Excessive oxidative stress may induce and promote breast carcinogenesis. Manganese superoxide dismutase (MnSOD) is critical to management of oxidative stress by catalyzing the formation of hydrogen peroxide from two superoxide anions. To examine the relationship between MnSOD Val-9Ala polymorphism, breast cancer and potential modifiers, we analyzed data from a large population-based case-control study. Study participants completed an in-home interviewer administered questionnaire, and self-completed a Block food frequency questionnaire. Age-adjusted unconditional logistic models included 1034 cases and 1084 controls. As compared with Val/Val genotype, we found no association between MnSOD Ala/Val (OR = 0.98, 95% CI: 0.79-1.21) and Ala/Ala (OR = 1.00, 95% CI: 0.79 1.28) genotypes and breast cancer. Results did not differ by menopausal status, stage of tumor, or estrogen and progesterone receptor status. No discernable patterns of interaction were found between this MnSOD variant and anti-oxidative exposures, including fruit and vegetable intake or NSAID use, or pro-oxidant exposures, including smoking and alcohol. This study provides little evidence that variation in Val-9Ala polymorphism of MnSOD alone or through substantial interaction with key exposures believed to be pro- or anti-oxidant properties influences breast cancer risk. PMID- 16215874 TI - Anthropometric factors and pancreatic cancer in a population-based case-control study in the San Francisco Bay area. AB - OBJECTIVE: To investigate the association between pancreatic cancer, anthropometric factors, physical activity and caloric intake. METHODS: Participants in our population-based case-control study of adenocarcinoma of the exocrine pancreas (532 cases, 1701 frequency-matched controls) in the San Francisco Bay Area were accrued between 1995 and 1999 and interviewed in-person. Data were analyzed by sex in age-adjusted unconditional logistic models and main effects were considered significant for two-sided p-values < or = 0.05. RESULTS: Odds ratios were elevated for body mass index (BMI) at age 25 years (4th versus 1st quartile: odds ratio (OR) = 2.0, 95% confidence interval (CI): 1.4-3.1), maximum BMI (OR = 1.8, 95% CI: 1.2-2.7) and usual adult BMI (OR = 2.1, 95% CI: 1.4-3.2) among men. Odds ratios were elevated for increased caloric intake among men (4th versus 1st quartile: OR = 2.6, 95% C: 1.7-3.8). Increased physical activity was suggestive of decreased risk in men and women although CIs included unity. Our results suggest that increased BMI and caloric intake are associated with pancreatic cancer among men. CONCLUSIONS: These results are consistent with other cancer studies and support further research to determine the mechanism by which increased BMI may influence the development of pancreatic cancer. PMID- 16215875 TI - Effects of a school-based obesity-prevention intervention on menarche (United States). AB - OBJECTIVE: Early menarche is a risk factor for breast cancer. Since body composition influences age at menarche we decided to estimate the effects of a school-based intervention for the prevention of obesity on the initiation of menses in young girls. METHODS: Ten schools were randomized to a modified curriculum or no curricular changes for 2 school-years. Data of 508 pre menarcheal girls at baseline (age range: 10-13 years) were analyzed. RESULTS: Girls attending an intervention school experienced menarche less frequently than girls attending control schools during the intervention period (intervention schools = 54%, control schools = 59%; RR = 0.76; 95% CI [0.66, 0.87]). Attending an intervention school was also associated with lower increase in BMI (-0.3 kg/m2; p = 0.003), lower gains in triceps skinfold thickness (-1.5 mm; p = 0.007), decreased television viewing (-0.6 h/day; p<0.0001) and increased physical activity (3.1 MET-h/week; p = 0.032). Including these changes as predictors of menarche incidence attenuated the intervention effect (RR = 0.94; 95% CI [0.80, 1.10]). CONCLUSIONS: The intervention delayed menarche in this group of girls. The delay was produced by increased physical activity, reduced television viewing and changes in BMI and fat distribution. These findings may have implications for the primary prevention of breast cancer. PMID- 16215876 TI - Occupational exposures and mycosis fungoides. A European multicentre case-control study (Europe). AB - OBJECTIVES: Mycosis Fungoides (MF) is a rare disease with an occurrence indicating that occupational exposures may play a role. To estimate the association between MF and occupational exposures as measured by means of an job exposure matrix (JEM). METHODS: A European multicenter case-control study was conducted from 1995 to 1997 and included seven rare cancers, one of which was MF. Patients between 35 and 69 years of age, diagnosed with MF (n=140), were recruited and the diagnoses were checked by a reference pathologist who classified 83 cases as definite, 35 cases as possible and 22 cases as not accepted. Among the 118 accepted cases, 104 cases were interviewed, of which 76 were definite cases. We selected population controls and colon cancer controls to serve all seven case groups. Altogether 833 colon cancer controls and 2071 population controls were interviewed. Based on the reported occupational experiences, a team of industrial hygiene specialists identified five potential exposures and developed an JEM. This JEM was used to estimate the odds ratios (OR) for MF as a function of these exposures. The JEM included aromatic and/or halogenated hydrocarbons (AAHs), chrome (VI) and its salts, electromagnetic radiations, silica and pesticides. RESULTS: Exposures to AHHs (OR 6.3, C.I 2.4 16.7 for male) were associated with a high MF risk. CONCLUSIONS: The study supports the hypothesis that some MFs have an occupational etiology but only a small fraction of exposed workers are apparently susceptible since the disease is so rare. PMID- 16215878 TI - The economics of integrated depression care: the University of Michigan study. AB - A goal of the Robert Wood Johnson Depression and Primary Care Initiative at the University of Michigan is to create and implement the clinical care and financial systems necessary to enable links between primary care and mental health specialty depression care. This paper describes the economic issues related to resources required, the mechanisms to distribute those resources, and the support that must be garnered from stakeholders. By systematic measurement and application, we assess the cost, price and selected consequences of these efforts. The study illustrates the need for both centralized and distributed capacity and support for innovative models of care. PMID- 16215877 TI - Trends in in-patient hospital utilization and surgical procedures for breast, prostate, lung and colorectal cancers in Canada. AB - OBJECTIVE: To analyse population-based trends of in-patient surgical procedures for breast (female), prostate, lung and colorectal cancers. METHODS: The Hospital Morbidity Files supplied hospital data and the Canadian Cancer Registry, incidence data. Age-adjusted rates were standardized to the 1991 Canadian population. RESULTS: All four cancers showed major changes in trends of surgical procedures. For breast cancer, the rate of in-patient breast conservation surgery (BCS) increased from 1981 to the early 1990s while the rate of mastectomy decreased. Because day surgery was not included, the subsequent in-patient BCS rate stayed level. For prostate cancer, the rate of transurethral prostatectomy was initially high but decreased after 1990, while the rate of radical prostatectomy increased rapidly, only minimally affected by the PSA-related peak in incidence. The lung cancer lobectomy rate in men remained at 10/100,000 after 1986, but in women rose from 3/100,000 to 7/100,000, reflecting increasing lung cancer incidence. For colorectal cancer, right hemicolectomies and anterior resections increased, especially in men. CONCLUSIONS: Surgery trends reflected changes in incidence and treatment preferences. Canadian trends were generally similar to US trends, although the timing of some of the changes differed. Canadians tended to use less invasive procedures such as BCS and anterior resection. PMID- 16215879 TI - Public attitudes toward psychiatric treatment. An international comparison. AB - AIM: In order to examine whether there is a relationship between the state of mental health care and the acceptance of psychiatry, public attitudes toward psychiatric treatment in three countries where the reform of mental health care has progressed to a different degree will be compared. METHODS: Population surveys on public beliefs about mental illness and attitudes toward psychiatric treatment were conducted in Bratislava, Slovak Republic, and Novosibirsk, Russia. The data were compared with those from a population survey that had recently been carried out in Germany. In all three surveys, the same sampling procedure and fully structured interview were applied. RESULTS: Although respondents from all three countries were equally inclined to seek help from mental health professionals, those from Bratislava and Novosibirsk tended to recommend more frequently to address other medical or nonmedical professionals or members of the lay support system. In all three countries, psychotherapy was the most favored treatment modality, followed by psychotropic medication. Although natural remedies were more frequently recommended in Bratislava and Novosibirsk, meditation/yoga was more popular among the German public. Across all three countries, the endorsement of a brain disease as cause was associated with a greater willingness to seek help from medical professionals (psychiatrist, GP). Respondents who adopted biological causes tended to recommend psychotropic medication more frequently. CONCLUSION: In countries with less developed mental health care systems, there appears to be a tendency of the public toward more frequently relying on helping sources outside the mental health sector and on traditional "alternative" treatment methods. However, it is our prognosis that with the progress of reforms observed, differences may further decrease. PMID- 16215880 TI - Methods of extracorporeal liver support for treatment of liver cell failure. PMID- 16215882 TI - Albumin-bound substances--a new target in liver failure therapy. PMID- 16215883 TI - Is it worth removing albumin-bound substances in hepatic encephalopathy? PMID- 16215884 TI - Acute alcoholic hepatitis. PMID- 16215885 TI - Mars in decompensated alcoholic liver disease with multi-organ failure. PMID- 16215886 TI - Successful clinical application of extracorporal albumin dialysis in a patient with benign recurrent intrahepatic cholestasis (BRIC). AB - This is a case report of a 36 years old man who has been suffering for 20 years from benign recurrent intrahepatic cholestasis (BRIC). BRIC is a rare autosomal recessive disease characterised by prolonged episodes of intrahepatic cholestasis and pruritus alternating with periods of nearly normal liver function, and does not progress to cirrhosis. Since all former approaches to medical treatment of the patients severe pruritus were ineffective, the patient was treated by 3 sessions of albumin dialysis (MARS, Molecular Adsorbents Recirculating System). MARS dialysis decreased serum bilirubin levels by more than 60 % and effectively lowered serum bile acid levels by 45 %. The course of serum parameters was accompanied by a dramatic clinical improvement of the patients symptoms (pruritus, jaundice, fatigue etc.). MARS therapy appeared to shorten the duration of the cholestatic attack. PMID- 16215887 TI - Artificial liver support as a bridge to orthotopic liver transplantation in a case of acute liver dysfunction on non-alcoholic steatohepatitis (NASH). PMID- 16215889 TI - First experience in Spain with MARS in three patients with advanced liver failure. PMID- 16215888 TI - Treatment of split-liver recipients with poor graft function by albumin-dialysis (Mars). PMID- 16215890 TI - Preliminary results of a phase I trial evaluating an extracorporeal hepatic support device utilizing albumin dialysis. PMID- 16215891 TI - Albumin binding capacity (ABiC) is reduced in commercially available human serum albumin preparations with stabilizers. PMID- 16215892 TI - Influence of albumin dialysis (MARS) on neuronal network activity in vitro--early results. PMID- 16215894 TI - Drugs in liver disease and during albumin dialysis -MARS. PMID- 16215918 TI - Mild postnatal anemia: is it a problem? AB - Mild postnatal anemia is common. However, determination of postnatal hemoglobin level or iron supplementation are not routine in many obstetric units. This study was designed to evaluate the impact of mild postnatal anemia and iron supplementation on women. One hundred fifty women who had postnatal hemoglobin levels between 80 and 99 g/L were randomly assigned into two groups with iron tablets or placebo provided from the time of diagnosis until reassessment at 6 weeks. The patients' general well-being, hemoglobin levels, iron status, side effects, and compliance were assessed. One hundred twenty-two women returned for reassessment. Twelve (9.8%) of the 122 women recruited and four (3.2%) of 122 nonanemic matched controls complained of dizziness 2 days after delivery; the difference was statistically significant (chi2 test, p<0.05). At 6 weeks postpartum, significantly more women were anemic (chi2 test, p<0.05) and iron deficient (chi2 test, p<0.01) in the placebo group. The score of general well being as assessed by a 4-point scale was significantly higher in the iron supplementation group (Mann-Whitney test, p<0.05). The amounts of drugs consumed in both groups were comparable and the incidences of side effects were similar. PMID- 16215919 TI - Prenatal diagnosis of cri du chat syndrome with encephalocele. AB - A 19-year-old primigravida was found to have an encephalocele on screening ultrasound study. Amniocentesis indicated cri du chat syndrome, 5p-. Although cri du chat syndrome has been noted in association with central nervous system malformations, encephalocele is a rare finding in this syndrome. PMID- 16215920 TI - Necrotizing enterocolitis in a premature infant as the presenting symptom of familial dysautonomia in the neonatal period: case report and review of the literature. AB - We present a case of a premature infant with early-onset necrotizing enterocolitis as the presenting symptom of familial dysautonomia. The diagnosis of familial dysautonomia in the neonatal period is usually rare and difficult, because many of the symptoms may be mild or nonspecific, whereas other characteristic signs and symptoms appear only later in life. The neonatologist should be aware of this rare diagnosis in the neonatal period, especially in the offspring of Ashkenazi Jews. The diagnosis is genetic. Preconception counseling to future parents belonging to populations at risk should include recommendation for genetic screening test to identify carriers of the mutations in the familial dysautonomia gene. PMID- 16215921 TI - Clinical use of new-generation pulse oximeters in the neonatal intensive care unit. AB - Continuous monitoring by pulse oximetry is a common practice for preterm and critically ill newborns. A new generation of motion-tolerant pulse oximeters have been designed for improved clinical performance with a substantial reduction in alarm frequency. However, little is known about the differences among these new generation pulse oximeters in the neonatal intensive care unit (NICU). The purpose of this study is to assess the clinical performance of two new-generation pulse oximeters in the NICU. Two new-generation pulse oximeters were used simultaneously to monitor 36 patients in the NICU. The two devices studied were the Philips FAST and the Masimo SET. Patients were randomly assigned for their digit selection and data were collected only when waveforms were of good quality and/or the pulse oximeter's pulse rate (PR) correlated with the electrocardiogram heart rate (HR). The data for oxygen saturation measurements, number of true and false alarms, and number of dropouts as well as the duration of dropouts for each pulse oximeter were recorded by the primary investigator at 5-minute intervals for a period of 2 hours on each patient. Dropouts are instances when the pulse oximeter alarm sounds due to its inability to identify the arterial pulse and provide an oxygen saturation reading. The mean gestational age for the study group was 32 weeks (rang, 24 to 42 weeks). Repeated-measures analysis of variance indicated no difference between the two devices across all time measurements (p=0.357). In addition, paired t-tests for true alarms and false alarms were not significant, with p-values of 0.151 and 0.869, respectively. There was a difference in the number of data dropouts (p<0.001): the Philips device had a nearly six-fold increase in the number of dropouts (Philips 247 versus Masimo 38). The duration of dropouts was also significant; the Philips device had three times longer duration of dropouts. Physiologic monitoring in the critical care setting requires accurate data measurements. The two new-generation pulse oximeters, the Philips FAST and Masimo SET, are equally sensitive in their ability to identify true and false alarms. Masimo pulse oximeter with its signal extraction technology, however, provides more consistent and accurate reporting of SpO2 values as demonstrated by its markedly decreased incidence of data dropouts. The Masimo unit appears to be more resistant to the effects of motion artifact. PMID- 16215922 TI - Bubbles in the brain veins as a complication of daily management of a scalp vein catheter. AB - Recently, it has been suspected that venous dural sinus air embolisms could occur as a result of scalp vein infusion. The possible mechanism for this complication has not been documented to date, and procedures to avoid venous dural sinus air embolism have not been presented. We report a preterm neonate who demonstrated venous dural sinus air embolism following daily management of a scalp vein catheter. The air embolism disappeared in a few days without complications. Clinicians should avoid injecting air into a scalp vein catheter and leaving it open to air. Following careful examination and review of the infant's case, we were able to conclude that positioning the infant in either supine or Trendelenburg position during daily management of scalp venous infusions and using an air-occlusive dressing once the catheter is removed could limit the risk of venous dural sinus air embolisms caused by scalp vein infusions. PMID- 16215923 TI - Bart syndrome with associated anomalies. AB - Bart syndrome is an inherited condition characterized by epidermolysis bullosa and congenital absence of skin. It has been associated with other anomalies including pyloric atresia. The genetic abnormality has been linked to chromosome 3, with an autosomal dominant pattern of inheritance. We present a case of Bart syndrome that was associated with pyloric atresia. The literature is reviewed pertaining to this unusual association. Recommendations are offered regarding genetic counseling and anticipatory guidance for affected families. PMID- 16215924 TI - Cytomegalovirus seroprevalence among women of childbearing age during a 10-year period. AB - The incidence of congenital cytomegalovirus (CMV) disease is dependent on the rate of virus acquisition by susceptible women in temporal proximity to pregnancy. Information about serostatus may help to identify high-risk groups and target interventions. Each year from 1991 to 2000, cord blood specimens (surrogates for maternal serum) from 100 consecutive deliveries were obtained at three hospitals. The presence of CMV immunoglobulin G antibody was correlated with demographic data. Overall, 58% of 2992 women were seropositive. After stratification by socioeconomic status (SES), nonwhite race was strongly associated with seropositivity (odds ratio, 3.0; 95% confidence interval [CI], 2.5 to 3.8), and after stratification by race, lower SES also was associated (odds ratio, 2.0; 95% CI, 1.7 to 2.3). There were no trends in seroprevalence over time in any demographic group. However, among white women 21 to 29 years of age, a significant association was found with later birth cohort. Seropositivity ranged from 40% among older, white primigravidas of upper SES to 89% among older, nonwhite, multigravid women of lower SES. These data suggest high rates of CMV transmission among nonwhite women of lower SES. Because these women tended to deliver infants at younger ages, delaying pregnancy even a few years might reduce the incidence of congenital disease. In contrast, most white, upper SES women remain susceptible to primary infection during their childbearing years. Such women born after 1975 may be more likely to reach the childbearing years with CMV immunity as compared with women born before then. PMID- 16215925 TI - Cesarean section, gestational age, and transient tachypnea of the newborn: timing is the key. AB - The purpose of this study was to identify risk factors and to characterize infants with transient tachypnea of the newborn (TTN). A total of 67 newborns with TTN, born at gestational age (GA)>or=35 weeks, were studied. Newborns delivered before and after each study case served as controls. Mean GA was lower and cesarean section (CS) rate was higher in the TTN group (38.2+/-2.3 versus 39.5+/-1.4 weeks, p<0.001; 50.7% versus 22.4%, p<0.001). GA<38 weeks was found to be associated with increased risk for TTN in infants delivered by elective CS. TTN was associated with significant morbidities and longer hospital stay (7.2+/ 5.6 versus 2.9+/-1.4 days; p<0.001). Delivery by CS and younger GA are risk factors for TTN. Although TTN is a self-limited disease, it is associated with significant morbidities. Scheduling elective CS at GA of not less than 38 weeks may decrease the frequency of TTN. PMID- 16215926 TI - Peripartum disseminated extrapulmonary tuberculosis simulating ovarian carcinoma. AB - Disseminated extrapulmonary tuberculosis is an uncommon complication of pregnancy. We present a 26-year-old multiparous immigrant from Haiti who was admitted following an extramural preterm delivery. Marked ascites was confirmed by computerized tomography, which also revealed a thickened greater omentum. These findings were considered suggestive of advanced ovarian carcinoma, although extrapulmonary tuberculosis was also considered despite negative tuberculin skin test screening. Image-guided omental biopsy demonstrated caseating granulomas substantiating the diagnosis of abdominal tuberculosis, which was later confirmed by cultures. The patient responded well to antituberculosis medications. This case describes the unusual peripartum presentation of abdominal tuberculosis simulating advanced ovarian carcinoma, and demonstrates the importance of considering extrapulmonary tuberculosis when encountering ascites and omental thickening during pregnancy despite negative tuberculin skin test screening. PMID- 16215927 TI - Successful perinatal management of a very low birthweight infant with congenital complete atrioventricular block. AB - We report a very low birthweight infant with congenital complete atrioventricular block (CCAVB) who underwent delayed implantation of a permanent pacemaker without temporary pacing. The female infant was born at 30 weeks gestation and weighed 1422 g. After birth, the infant showed respiratory failure due to pleural effusion and respiratory distress syndrome at birth. The heart rate ranged between 50 and 55 bpm. The chest x-ray demonstrated dilated heart, but echocardiogram showed good systolic ventricular function. Respiratory failure was resolved after supportive treatment without temporary pacing. Mild heart failure due to low heart rate persisted, but was successfully managed by conventional heart failure therapy combined with nasal continuous positive airway pressure. She achieved a body weight gain to 1856 g at the age of 49 days and underwent implantation of a permanent pacemaker. We conclude that it is important to determine the timing of both delivery and pacemaker implantation for successful perinatal management of infants with CCAVB. PMID- 16215928 TI - Trends in mode of delivery for gastroschisis infants. AB - Cesarean delivery for infants with gastroschisis has not been clearly demonstrated to produce superior outcomes compared with vaginal delivery; however, it not clear whether this has been translated into common clinical practice. Therefore, we analyzed the shifts in type of delivery during the last three decades at our institution. The medical records of all infants with gastroschisis treated at our hospital during the last 33 years were reviewed. Two groups, based on mode of delivery, were compared with regard to demographics, treatment, and outcome. A statistical analysis of changes in type of delivery was performed, and compared with the neonatal population as a whole. A total of 222 patients with gastroschisis were analyzed: 103 delivered via cesarean section and 119 delivered via vaginal birth. Logistic regression demonstrated a significant trend toward cesarean delivery over time (p=0.0001). The odds ratio indicated a 16% per year increase in the chance of cesarean delivery. There has been a significant trend toward cesarean delivery in patients with gastroschisis treated at our institution. PMID- 16215929 TI - Aortic dissection in a neonate: case report and review of the literature. AB - We report a case of aortic dissection causing hypertension in a neonate, which occurred following iatrogenic intimal injury during umbilical arterial cannulation. The intimal flap was diagnosed by color Doppler sonography, and treated by conservative management with complete spontaneous healing of the intimal injury. PMID- 16215930 TI - Cloning of full-length cDNA sequence of a novel candidate gene related to mouse cleft palate. AB - OBJECTIVE: Cloning and screening a novel candidate gene related to developing mouse cleft palate. METHODS: The differentially expressed genes were cloned by modified PCR-based subtractive hybridization. After identification using reverse dot blotting, positive clones were sequenced and analyzed for homology in GenBank databases. RESULTS: Four novel express sequence tags were obtained, one of which spanning 809 bp was the full-length of the novel gene cDNA identified by Northern hybridization. CONCLUSION: A novel candidate gene related to mouse cleft palate was cloned. PMID- 16215931 TI - Effect of polymorphism of uncoupling protein 3 gene -55 (C>T) on the resting energy expenditure, total body fat and regional body fat in Chinese. AB - OBJECTIVE: To investigate the relationship of the C to T variant at the -55 site of the promoter region of uncoupling protein 3 gene (UCP3) with the resting energy expenditure and the parameters of body fat in Chinese population. METHODS: Three hundred Chinese (91 normal weight subjects, 209 overweight/obesity subjects) were genotyped for the UCP3 gene -55(C>T) by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Resting energy expenditure (REE), fat mass (FM), fat free mass (FFM) and the parameters for regional adipose tissue distribution were measured. RESULTS: Genotype frequencies of UCP3 gene -55(C>T) were not associated with obesity and different types of obesity. The REE level in normal weight subjects with TT homozygotes was higher than that in those with CT heterozygotes and CC homozygotes (P=0.0200). Similar tendency was also observed in overweight/obesity subjects. The FM/FFM exhibited significant difference between the overweight/obesity subjects with a TT genotype and those with a CT or CC genotype (P=0.0096). CONCLUSION: The level of difference in REE caused by the polymorphism of promoter region of UCP3 -55(C>T) may play a role in energy metabolism in Chinese. PMID- 16215932 TI - Novel GLA gene mutations in two Chinese families with classic Fabry disease. AB - OBJECTIVE: To search mutations in GLA gene in two Chinese families with classic Fabry disease. METHODS: Two families with Fabry disease confirmed by pathological and clinical studies were reported here. In pedigree 1, 12 family members had paroxysmal pain on limb extremities. In pedigree 2, there were 8 patients and most of them had multi-organ involvement at the end stage of the disease. Two probands from the two families together with several of their family members were searched for mutations in GLA gene. After extraction of genomic DNA from peripheral leukocytes, all of the 7 exons and their flanking introns were amplified by PCR and directly sequenced. RESULTS: Both the proband 1 and proband 2 were identified to be hemizygotes of novel GLA missense mutations. G132T (TGG- >TGT) mutation in exon 1, resulting in the substitution of amino acid from tryptophan to cysteine (W44C), was detected in proband 1. G874C (GCT-->CCT) mutation in exon 6, resulting in the substitution of amino acid from alanine to praline (A292P), was detected in proband 2. Mothers of the 2 probands were heterozygotes carrying the same mutation as their sons. CONCLUSION: We report here 2 novel missense mutations in two Chinese families with classic Fabry disease. Different mutations in the same gene can result in phenotypes with significant deviation. Several female patients with the same clinical manifestations as male patients in the 2 families suggest that the X-linked dominant inheritance of the disease, possibly related to be the random X chromosome inactivation. PMID- 16215933 TI - A minidystrophin-EGFP fusion gene expressed in Cos-7 cells mediated by human source vector. AB - OBJECTIVE: To construct a human source vector containing minidystrophin-EGFP fusion gene and investigate its expression in Cos-7 cells. METHODS: The recombinant human source vector named pHrnDysG was constructed with PCR-clone methods. Three fragments of dystrophin gene were PCR amplified from normal human dystrophin gene cDNA (GenBank NM04006). These three fragments were ligated to generate a minidystrophin gene. The enhanced green fluorescent protein (EGFP) gene was fused to the C terminal of the minidystrophin gene, and then the pHrnDysG was finally obtained by cloning the fusion gene to pHrneo. Fluorescence microscope and RT-PCR were used to detect the expression of minidystrophin-EGFP fusion gene after the recombinant construct was transfected into Cos-7 cells by lipofectamine. RESULTS: Restrictive enzyme digestion analysis and sequencing confirmed that pHrnDysG vector was constructed successfully. After the recombinant pHrnDysG was transfected to Cos-7 cells, RT-PCR demonstrated that the fusion gene was successfully transcribed, and the green fluorescence was observed at the cell membrane. CONCLUSION: The minidystrophin-EGFP fusion gene mediated by pHrneo vector could express in Cos-7 cells and its products' localization in the cell membrane was the same as that of full length dystrophin. These results suggested that the recombinant human source vector pHrnDysG might be potentially used in studies on the gene therapy of Duchenne muscular dystrophy. PMID- 16215934 TI - Analysis of single nucleotide polymorphisms and haplotypes in HOXC gene cluster within susceptible region 12q13 of simple congenital heart disease. AB - OBJECTIVE: In the candidate region 12q13 of simple congenital heart disease(CHD), four single nucleotide polymorphisms(SNPs) in HOXC4 gene were chosen in order to investigate the distribution of SNP and haplotypes in simple CHD patients and normal people. METHODS: The genotype of 4 SNPs in 108 simple CHD patients and 200 normal people were analyzed by restriction fragment length polymorphism(RFLP) and denaturing high-performance liquid chromatography(DHPLC). The statistical contingency table method was used to analyze SNP genotype frequency and gene frequency in patients and control group; then, the haplotypes were established and their frequencies in the two groups were assessed by PHASE software. RESULTS: C16476T polymorphism was not detected; A17860G located in 3' flanking sequence of HOXC5 gene displayed significant difference between the two groups. The G allele frequency in simple CHD patients was higher than that in healthy controls(P < 0.05); the distribution of frequencies of 4 haplotypes showed significant difference(P < 0.01). CONCLUSION: The A17860G located in 3'flanking sequence of HOXC5 gene is associated with simple CHD; the risk of CHD in the persons with G17860 is higher than that in those with A17860. the haplotype of 3 SNPs may be linked with the susceptible gene of simple CHD. PMID- 16215936 TI - Mutations of 15 short tandem repeat loci in Chinese population. AB - OBJECTIVE: To explore the mutations of 15 short tandem repeat (STR) loci in PlowerPlex16 System which are world-widely used in parentage testing. METHODS: Mutations of 15 STR loci in PlowerPlex16 System were investigated in 1921 parentage testing cases from Chinese population. RESULTS: In 1921 parentage cases, seventy cases (3.644%) were found to have mutations. Among these were one case with double mutations (D21S11 and PentaD) and another case with two different mutations (D7S820 and D16S539) in two children. The total number of mutated STR loci observed was 72 over 3764 meiosis with a mutation rate of 0.128% +/- 0.1104% x 10(-3). The highest mutation rate was 0.292% at vWA and D21S11. No mutation was observed at TH01 or at TPOX. The mutated alleles coming from father were five times more than those from mother. The majority (98.611%) of mutated alleles were the results of one-step mutation. The ratio of one-step gain versus loss was 1.826:1. There was only one multiple-step mutation with a double-repeat gain observed at PentaD locus. In the PlowerPlex16 System, nine loci, namely D8S1179, Penta D, D13S317, D16S539, D7S820, D5S818, D3S1358, TH01 and TPOX, have lower mutation rates and are more suitable for parentage testing. CONCLUSION: Mutation of STR is relatively common and often makes parentage testing more complicated. Selecting stable STR locus with low mutation rate is more important in parentage testing. PMID- 16215935 TI - The construction, characterization and transfection of liposome- polycation CDKN1B plasmid complexes. AB - OBJECTIVE: To develop an efficient non-viral gene delivery system in order to transfer CDKN1B gene efficiently into lung and liver carcinoma cells. METHODS: A recombinant plasmid composed of CDKN1B sequence and EYFP as reporter gene was constructed and identified. The recombinant DNA was then formulated the lipids polycation-DNA complexes(LPDs) with protamine sulfate. Several kinds of lung and liver carcinoma cells were transfected by means of LPDs. The physicochemical properties of LPDs were investigated using PCS method and TEM, respectively. The expression of EYFP in A549 cells was observed under fluorescent microscope and evaluated by flow cytometry analysis. Finally, the production of CDKN1B protein in transfected LLC, Chang and 7721 cells was identified by Western blot analysis. RESULTS: The average diameter of the LPDs were 167 nm with the polydispersity index of 0.35. The average zeta potential of LPDs was +32.6 mV. LPDs look like a sunken sphere. The fluoresent microscope picture clearly indicated the expression of EYFP in A549 cells. The flow cytometry result showed that the transfection efficiency of LPDs in A549 cells was comparable with that of LipofectAMINE, the positive control. Western blot analysis confirmed the production of CDKN1B protein in LLC, Chang and 7721 cells transfected with LPDs, while no CDKN1B protein was detected in cells transfected with naked DNA. CONCLUSION: The construction of the recombinant plasmid is successful. LPDs can deliver the recombinant plasmid to lung carcinoma cells and liver carcinoma cells with high efficiency. Therefore, this kind of gene delivery system has the potential uses for the treatment of lung and liver cancer. PMID- 16215937 TI - [Mutation analysis of small heat shock protein 27 gene in Chinese patients with Charcot-Marie-Tooth disease]. AB - OBJECTIVE: To investigate the features of small heat shock protein 27 (HSP27) gene mutation in Chinese patients with Charcot-Marie-Tooth disease (CMT). METHODS: DNA samples from 114 CMT probands were screened for mutations in HSP27 gene by polymerase chain reaction and direct sequencing, and haplotype analysis was further carried out on the mutation detected families. RESULTS: One missense mutation C379T was detected in 4 autosomal dominant CMT2 families. Haplotype analysis indicated that the 4 families probably had a common ancestor. CONCLUSION: To the authors' knowledge, this is the first report of HSP27 gene mutation in Chinese patients with CMT, but it may be not common(0.90%). The C379T mutation in HSP27 gene also causes CMT2 except for distal hereditary motor neuropathy, thus providing further evidence that even the same mutation in the same gene may lead to distinct phenotypes. PMID- 16215938 TI - [A study on PARKIN gene in three pedigrees with autosomal recessive early-onset Parkinson's disease]. AB - OBJECTIVE: To detect the possible relationship between PARKIN gene and the Chinese pedigree with autosomal recessive early-onset Parkinson's disease(AREP). METHODS: Clinical examination was carried out in 6 patients from 3 Chinese pedigrees with AREP and their 23 family members. PCR amplification of all exons of PARKIN gene was performed. The PCR products were analyzed by denaturing high performance liquid chromatography(DHPLC) to screen for point mutation and polymorphism. And in the samples with abnormal DHPLC result, further sequencing was conducted to confirm the type of mutation and polymorphism. RESULTS: All exons of PARKIN gene from the research subjects were successfully amplified. A heterozygous point mutation (Gly284Arg) in exon 7 was found in one pedigree. A polymorphism (Ser167Asn) in exon 4 was found in another pedigree. All the patients had the past history of exposure to environmental poison. CONCLUSION: When acting together with risky environmental factors, the heterozygous mutation Gly284Arg in PARKIN gene may cause AREP. The polymorphism Ser167Asn in PARKIN gene increases the risk of developing Parkinson's disease and may cause AREP when acting together with hydrargyrism. PMID- 16215939 TI - [Identification and characterization of a novel gene EOLA1 stimulating ECV304 cell proliferation]. AB - OBJECTIVE: To amplify the full-length cDNA and characterize the structure and biological function of a novel expression sequence tag ST55 (GenBank Accession No. BM121646). METHODS: Rapid amplification of cDNA ends was used to clone the full-length of cDNA of ST55 in this study. Then, its tissue distribution was checked by Northern blots, and the associated protein was screened by GAL 4-based yeast two-hybrid. The effect of stable transfection of the cDNA on cell proliferation was evaluated in ECV304 cells. RESULTS: A full-length 1404 bp cDNA was cloned, and it was accepted as a novel human mRNA by GenBank (No. AY074889), named endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1). Bioinformatic analysis found that the EOLA1 encoded 158 amino acids, 17.89 kDa protein, and mapped to chromosome Xq27.4 with 5 exons. EOLA1 expressed in different human normal tissues and cancer cell lines. Using the EOLA1 cDNA as bait, we performed a yeast two-hybrid screening of a human liver cDNA library and identified metallothionein 2A (MT2A) as associated protein. The interaction between EOLA1 and MT2A was confirmed by co-immunoprecipitation experiments. Stable transfection of EOLA1 was noted to stimulate ECV304 cell proliferation (P < 0.05). CONCLUSION: The findings suggest that EOLA1 is a novel gene and the interaction of EOLA1 and MT2A may play an important role in cell protection in inflammation reaction. PMID- 16215940 TI - [Sex chromosome analysis and IGF-II expression on activated human unfertilized oocytes after ICSI with calcium ionophore A23187 and puromycin]. AB - OBJECTIVE: To investigate the sex chromosomes and the expression of insulin-like growth factor-II (IGF-II) on activated human unfertilized oocytes after intracytoplasmic sperm injection(ICSI) with calcium ionophore A23187 and puromycin. METHODS: All 95 discarded oocyes that showed no evidence of fertilization at 16-18 h after in vitro maturation and intracytoplasmic sperm injection cycles (IVM-ICSI)/conventional ICSI were exposed to calcium ionophore A23187 (5 micromol/L) for 5 min and then were incubated with puromycin (10 microg/mL) for 4 h. After activation, the oocytes were cultured in vitro for 3-5 days. The sex chromosome analysis was performed by dual color fluorescence in situ hybridization. The expression of IGF-II on the activated embryos, normal embryos, and parthenotes was examined. RESULTS: The combination of calcium ionophore A23187 with puromycin could activate the unfertilized oocytes 22 h after ICSI. The activated rate, cleavage rate, and quality of activated embryos of the IVM-ICSI group were similar to those of ICSI group, respectively. Sex chromosome analysis indicated that 8 male and 5 female embryos had been derived from two pronucleus and a second polar body. The expression of IGF-II on activated embryos and normal embryos was high and similar, which was much stronger than that of parthenotes. CONCLUSION: The combination of calcium ionophore A23187 with puromycin could effectively activate unfertilized oocytes 22 h after ICSI. Moreover, the unfertilized oocytes activated by calcium ionophore A23187 and puromycin had normal sex chromosomes and expression of IGF II like the normal embryos. These suggest that oocyte activation may be considered as a remedial measure in the presence of total or nearly total fertilization failure in ICSI. PMID- 16215941 TI - [Relationship of interleukin-18 gene promoter polymorphisms with chronic hepatitis B in Chinese Han population]. AB - OBJECTIVE: To investigate the polymorphisms of interleukin-18(IL-18) gene promoters, and to disclose whether such polymorphisms are associated with susceptibility to chronic hepatitis B in Chinese Han population. METHODS: Using polymerase chain reaction with sequence specific primers method, the authors detected the single nucleotide polymorphisms of the promoter region of IL-18 gene at position -607C/A and -137G/C in 231 patients with chronic hepatitis B and 300 normal controls. RESULTS: The frequency of CC genotype in IL-18 gene promoter region at position -607 was 0.22(66/300) in normal controls and 0.27(62/231) in patients. The frequency of CA genotype was 0.53(160/300) in normal controls and 0.50(116/231) in patients. The frequency of AA genotype was 0.25(74/300) in normal controls and 0.23(53/231) in patients. The frequencies of -137GG, GC and CC genotype were 0.67, 0.30 and 0.03 in normal controls respectively; whereas in chronic hepatitis B patients the frequencies were 0.79, 0.19 and 0.02. The genotype frequency of -137GG in chronic hepatitis B groups was significantly higher than that in normal controls(chi2: 8.55, P=0.003), but the frequencies of 607C/-137C and -607A/-137C haplotypes in chronic hepatitis B groups were significantly lower than those in normal controls. The association between genotype of IL-18 promoter region polymorphisms and hepatitis B virus(HBV) copies showed that the frequency of -607AA genotype in high HBV-DNA copies groups was lower than that in low HBV-DNA copies groups(chi2: 6.03, P=0.014). CONCLUSION: The polymorphisms of the promoter region of IL-18 gene at position -607C/A and 137G/C are correlated with chronic hepatitis B in Chinese Han population. The people with -137C allele in the promoter region of IL-18 gene may be protected against HBV infection, and the IL-18 -607AA genotype may be linked to HBV-DNA copy. PMID- 16215942 TI - [Association study of sleep apnea syndrome and polymorphisms in the serotonin transporter gene]. AB - OBJECTIVE: To identify polymorphisms of the serotonin transporter(5-HTT) gene and to find out whether there was relationship between any such polymorphisms and sleep apnea syndrome (SAS). METHODS: For two polymorphisms of 5-HTT target DNA gene was amplified using polymerase chain reaction (PCR) and 6% non-denaturing polyacrylamide gels electrophoresis. The frequencies of the different forms of the genotypes and alleles of 5-HTT gene were analyzed in 104 patients with SAS and 150 healthy controls. RESULTS: The frequencies of the S or L alleles and the S/S, S/L or L/L genotypes in promoter region of 5-HTT gene in SAS group were not significantly different to those in healthy controls (P > 0.05). However, the frequencies of 10/10, 12/10 genotypes of 5-HTT-VNTR in SAS patients were significantly higher than those in healthy control subjects (P < 0.05). Moreover, the frequency of the allele 10 of 5-HTT-VNTR in SAS patients was significantly higher than that in healthy controls (P<0.01). CONCLUSION: The allele 10 of 5-HTT VNTR might be a susceptible factor in the pathogenesis of SAS. PMID- 16215943 TI - [Detection of duplications or deletions of the PMP22 gene using real-time quantitative PCR]. AB - OBJECTIVE: To detect the duplication or deletion of peripheral myelin protein 22(PMP22) gene in Chinese patients with Charcot-Marie-Tooth disease(CMT) or hereditary neuropathy with liability to pressure palsies(HNPP) using real-time quantitative polymerase chain reaction. METHODS: Duplications or deletions of PMP22 gene were detected in 113 CMT cases, 4 HNPP cases and 50 normal controls by using real-time quantitative PCR. RESULTS: Thirty-six of 113 CMT cases had the PMP22 duplication, 4 HNPP cases had the PMP22 deletion. No duplication or deletion was found in 50 normal controls. CONCLUSION: The PMP22 duplication rate in Chinese patients with CMT is 31.9%(36/113). PMP22 deletion is the common cause of HNPP. PMID- 16215944 TI - [Generation of transgenic mice expressing human lysozyme in mammary gland]. AB - OBJECTIVE: To evaluate the feasibility of generating animal mammary gland bioreactors expressing human lysozyme (hLYZ). METHODS: The recombinant vector p205C3-hLYZ, as a result of connecting the hLYZ cDNA with the mammry gland expression vector p205C3, was used to generate transfer genic mice by microinjection. RESULTS: A total of 136 F0 mice were obtained, of which 7 (2 females and 5 males) and 4 (1 females and 3 males) were found to contain the transfer-gene by PCR and Southern blotting respectively. The results of Western blotting indicated that the expressed protein had the same molecular weight as that of normal hLYZ. From the F1 generation on, the mice mated only with their brothers or sisters and a colony of F7 transgenic mice was obtained. Among the offspring, the female transgenic mice maintained and expressed the transfer-gene stably with an expression level as high as 750 mg/L. The expressed protein had strong tissue specificity, and in addition to the mammary glands, some degree of ectropic expression in the spleens and intestines of the transgenic mice was confirmed by dot blotting assay. CONCLUSION: These data indicate that the mice mammary gland bioreactors expressing hLYZ have been successfully generated. PMID- 16215945 TI - [BCL-2/IgH translocation in peripheral blood cells of healthy Chinese individuals of Han nationality located in Zhejiang area]. AB - OBJECTIVE: To investigate the relationship between the frequency of BCL-2/IgH rearrangement in peripheral blood cells of healthy Chinese individuals of Han nationality located in Zhejiang area and the low incidence of follicular lymphoma (FL). METHODS: Nested-PCR and direct DNA sequencing were used to detect the Bcl 2/IgH rearrangement in peripheral blood cells of 196 healthy individuals. DNA sequences involved were then searched and aligned in NCBI database to confine the broken points in major breakpoint region and the IgH segments involved. RESULTS: First, in this sample the frequency of BCL-2/IgH translocation in Chinese individuals of Han nationality located in Zhejiang area is 9.66%, being much lower than that in North America and Europe countries. Second, the breakpoints tend to fall into 3 clusters: 3055, 3116 and 3165 bp. Usage of J6 segment is most common. Third, There are different subclones of BCL-2/IgH rearrangements in the same individual. CONCLUSION: The low frequency of BCL-2/IgH translocation in healthy Chinese individuals of Han nationality located in Zhejiang area may be one of the reasons for the difference in the incidence of FL between China and Western countries. PMID- 16215946 TI - [Detection of fusion genes resulting from chromosome abnormalities in childhood acute lymphoblastic leukemia]. AB - OBJECTIVE: To detect the expression of the fusion genes resulting from chromosome abnormalities in childhood acute lymphoblastic leukemia(ALL) and its conformity to WHO classification. METHODS: Sixty-two children with ALL were investigated. The expression of fusion genes was determined by multiplex reverse transcription polymerase chain reaction (RT-PCR), karyotyping (R band) and immunophenotyping (by flow cytometry) were also performed. RESULTS: Of the 62 patients, 23(37.1%) were found to carry 13 different fusion genes. The patients with immunophenotype of Pre-B-ALL were found to carry: TEL/AML1(3 cases); E2A/PBX1, E2A/HLF, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX-MLL/AF6-MLL/ELL, MLL/AF6-MLL/ELL, dupMLL (one case for each); and HOX11 (6 cases). The patients with immunophenotype of Pre-T ALL were found to carry: TAL1D (4 cases, one is also found to have HOX11 expression); and HOX11 (2 cases). The multiplex RT-PCR in combination with chromosome analysis revealed genetic abnormalities in 69.4%(43/62) of childhood ALL. CONCLUSION: Multiplex RT-PCR combined with chromosome analysis and immunophenotyping can provide reliable and helpful information for the diagnosis, therapy evaluation and prognosis prediction in childhood ALL, which may also serve as a basis on which to implement the criteria of WHO classification. PMID- 16215948 TI - [Association of -160(C-->A) polymorphism in CDH1 gene with gastric cancer risk in Fujian Chinese population]. AB - OBJECTIVE: CDH1, encoding E-cadherin, is an important tumor suppressor gene. The present study aims to investigate the association of -160(C-->A) polymorphism in CDH1 gene with susceptibility to gastric cancer in Fujian province. METHODS: One hundred and two patients from independent families and 101 healthy control subjects were analyzed. Genotype analysis was performed through polymerase chain reaction-based denaturing high performance liquid chromatography. The odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by unconditional logistic regression model. RESULTS: The frequencies of -160(C-->A) polymorphism CC, CA and AA genotype in case group and controls were 58(56.9%), 38(37.3%)i6(5.9%) and 55(54.5%), 41(40.6%), 5(5%), respectively. AA genotype did not present a significantly increased risk for gastric cancer (OR=1.12; 95% CI:0.32-3.95). No association was found between A allele and clinicopathological characteristics of gastric cancer. CONCLUSION: -160(C-->A) polymorphism in CDH1 gene promoter region may not be in association with genetic susceptibility to gastric cancer in Chinese population from Fujian. PMID- 16215947 TI - [A Chinese autosomal dominant polycystic kidney disease family probably related to PKD2 gene]. AB - OBJECTIVE: To study the genetic heterogeneity of autosomal dominant polycystic kidney disease (ADPKD) in Chinese. METHODS: Using polymerase chain reaction (PCR) and non-denatured polyacrylamide gel electrophoresis, the authors analyzed eight microsatellite markers closely linked to PKD1 or PKD2 genes respectively in a Chinese ADPKD family. RESULTS: Seven informative markers were found in this family, including KG8, SM6, CW4 and CW2 which are tightly linked to PKD1, and D4S1563, D4S414 and D4S423 which are linked to PKD2. After the process of genotyping, the haplotypes were estimated with Cyrillic 2.0, and the linkage based analysis suggested that the disease is not linked to PKD1 other than PKD2. CONCLUSION: In China this non-PKD1 family is the second one, but it is the first reported PKD2 family showing the genetic heterogeneity of ADPKD in Chinese. In the family the affected mother transmits the disease and the affected members' phenotypes are eterogeneous. In addition, the existing "anticipation" and the presence of the disease in a child of this family suggest that non-PKD1 linked families may have early-onset of the disease in child. PMID- 16215949 TI - [Using multiplex PCR to analyze the breakpoint of a severe Y-chromosome deletion]. AB - OBJECTIVE: To elucidate the relationship between azoospermia factor(AZF) microdeletion of Y chromosome and azoospermia, the exact breakpoint of a severe Y chromosome deletion was determined according to the physical map of AZF. METHODS: Multiplex polymerase chain reaction was used to amplify fifteen sequence tagged sites (STS), namely sY82, sY84, sY86 in AZFa, sY124, sY127, sY128, sY133, sY134, sY143 in AZFb, sY239, sY242 sY254, sY255 in AZFc, and sY145, sY152 in AZFd; sex determining region Y(SRY) was taken as an internal control. And then sY82,sY86,sY85,sY84 were further analyzed using the sample of the patient who had Y-chromosome deletion by G band analysis to map the breakpoint at molecular level. RESULTS: All 15 STS and sY85 were amplified in positive control while only sY82, sY86 were amplified in the clinical sample, thus the breakpoint was found to be between sY86 and sY85. CONCLUSION: This study on the patient provided the direct biomolecular evidence of the exact breakpoint of the severe Y-chromosome deletion and established the deletion map of acrocentric chromosome. It also proved that the patient's azoospermia was due to the deletion of AZF. PMID- 16215950 TI - [Preliminary analysis of retinal gene expression profile of diabetic rat]. AB - OBJECTIVE: Establishing the retinal gene expression profiles of non-diabetic rat and diabetic rat and comparing the profiles in order to analyze the possible genes related with diabetic retinopathy. METHODS: The whole retinal transcriptional fragments of non-diabetic rat and 8-week diabetic rat were obtained by restriction fragments differential display-PCR (RFDD-PCR). Bioinformatic analysis of retinal gene expression was performed using soft wares, including Fragment Analysis. After comparison of the expression profiles, the related gene fragments of diabetic retinopathy were initially selected as the target gene of further approach. RESULTS: A total of 3639 significant fragments were obtained. By means of more than 3-fold contrast of fluorescent intensity as the differential expression standard, the authors got 840 differential fragments, accounting for 23.08% of the expressed numbers and including 5 visual related genes, 13 excitatory neruotransmitter genes and 3 inhibitory neurotransmitter genes. At the 8th week, the expression of Rhodopsin kinase, beta-arrestin, Phosducinirod photoreceptor cGMP-gated channel and Rpe65 as well as iGlu R1-4 were down-regulated. mGluRs and GABA-Rs were all up-regulated, whereas the expression of GlyR was unchanged. CONCLUSION: These results prompt again that the changes in retinal nervous layer of rat have occurred at an early stage of diabetes. The genes expression pattern of visual related genes and excitatory and inhibitory neurotransmitters in rat diabetic retina have been involved in neuro dysfunctions of diabetic retina. PMID- 16215951 TI - [Effect of the mutation of promoter region in Wilson disease ATP7B gene on the expression of reporter gene]. AB - OBJECTIVE: To find out the relationship between mutation of ATP7B gene promoter region and pathogenesis of Wilson disease(WD). METHODS: Two of 48 WD patients presented C-->T base substitution mutations at the position -183. DNA sequences of the promoter region from normal and mutant samples were separated. The fragments containing the promoter region were cloned upstream of the luciferase. Luciferase activity was analyzed. RESULTS: The luciferase activity of reporter gene containing normal sequence of ATP7B gene promoter region did not show significant difference as compared with that of reporter gene containing mutant promoter(n=3, P > 0.05). CONCLUSION: No influence of C-->T base substitution mutations on the activity of promoter was observed in study. The results suggest that WD pathogenesis relates little to the mutations of the promoter region in Chinese. PMID- 16215953 TI - [Correlation between fibrinogen polymorphisms and the type of cerebral infarction]. AB - OBJECTIVE: To study the association of beta-fibrinogen(Fg) gene -148 C/T and 448 G/A polymorphisms, plasma Fg concentration, molecular reactivity and the type of cerebral infarction. METHODS: Gene polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism. The plasma Fg concentration and the molecular reactivity were also determined. RESULTS: The Fg concentration in MCI patients with T -148 allele was higher than that in PCI patients and controls. The MCI patients with A448 allele had higher Fg concentration, FMPV and FMPV/Amax when compared with controls, and had higher FMPV/Amax when compared with PCI patients. CONCLUSION: FgB beta -148 and 448 mutational genotypes have impact on Fg concentrationi and therefore increase the risk of MCI. PMID- 16215952 TI - [Association of angiotensin I converting enzyme 2 gene polymorphism with essential hypertension in Chinese]. AB - OBJECTIVE: To identify the genetic variants of angiotensin I converting enzyme 2 (ACE2) gene in a Chinese population and to determine whether the ACE2 gene polymorphisms are associated with essential hypertension (EH). METHODS: Seven hundred and forty-five patients with EH and 362 normal blood pressure controls were included in the study to assess the contribution of polymorphism of ACE2 gene. Direct DNA sequencing was performed to detect the single nucleotide polymorphisms (SNPs) in 20 subjects who were randomly selected from the EH patients. RESULTS: One SNP named G8790A located in the 4th base of the third intron was found in the 20 patients. The genotyping data indicate that the A allele frequency in male EH patients complicated with cardiac incompetence(55%) is significantly different from that in the control group(43.3%)(P<0.01). The A allele frequency in female patients with cardiac incompetence (56.1%) is higher than that in the controls (50.5%), but the difference does not reach statistical significance. CONCLUSION: The G8790A polymorphism may be related to the essential hypertension with cardiac incompetence in Chinese population. Additional investigation will be need to confirm the association. PMID- 16215954 TI - [No association of -1438G/A polymorphism in promoter region of 5-HT2A receptor gene with antipsychotic agent-induced weight gain]. AB - OBJECTIVE: To investigate whether the -1438G/A polymorphism in the promoter region of 5-HTR2A gene associates with the weight gain following antipsychotic agents (APS) acute treatment in schizophrenic patients. METHODS: Eighty-four Chinese Han patients with schizophrenia at the first onset were recruited from among 70 nuclear families. The polymorphism of 5-HTR2A gene was determined with PCR-RFLP technique. Body weight was measured in the patients on admission after 10 weeks of treatment with risperidone or chlorpromazine. RESULTS: There were no statistically significant differences in the distribution frequencies of genotype (chi2: 0.172, v1, P > 0.05) and allele (chi2: 0.121, v1, P > 0.05) of -1438G/A polymorphism of 5-HTR2A gene between subgroups (weight gain >or= 7% or < 7%). Likewise, there was no significant difference in weight gain between genotype groups. By means of transmission disequilibrium test and quantitative transmission disequilibrium test, no significant association between the -1438G/A polymorphism of 5-HTR2A gene and weight gain was observed. CONCLUSION: 5-HTR2A gene -1438G/A polymorphism was probably not associated with APS-induced weight gain in Chinese Han patients with schizophrenia in this study. PMID- 16215955 TI - [Polymorphism of DXYS267 locus and application of its Y-specific single nucleotide substitutions]. AB - OBJECTIVE: To investigate the polymorphism of DXYS267 locus in China Han population and find the application and characters of its Y-specific single nucleotide substitutions. METHODS: The locus was analyzed by PCR and PAGE in silver-staining. The Y-specific STR was amplified with newly designed primers according to the Y-specific single nucleotide substitutions. RESULTS: Six alleles were detected in Han population in Wuhan. Exact tests demonstrated that genotype frequencies did not deviate from Hardy-Weinberg equilibrium. Heterozygosity of DXYS267 was 0.6706, discrimination power (DP) was 0.8433, and the probability of paternity exclusion (PE) was 0.5957. The Y-specific STR of DXYS267 was successfully amplified with the new primer. The 4 alleles for Y-STR were detected with haplotype diversity (HD) 0.6372. CONCLUSION: The DXYS267 locus is appropriate for individual identification and paternity testing. The new primer is useful for individual and paternity testing involving brothers and mixed stains. PMID- 16215956 TI - [Comparison of Rhesus boxes in Hans and Uighurs]. AB - OBJECTIVE: To study the difference and similarity between Hans and Uighurs in regard to Rhesus box and its significance. METHODS: The sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed on the basis of RHD gene sequence. The upstream, downstream and hybrid Rhesus boxes were determined by polymerase chain reaction-sequence specific primer(PCP-SSP) and mismatched PCR. RESULTS: The percentage of RHD-/RHD-, RHD+/RHD- and RHD+/RHD+ genotypes ascertained in the unrelated Hans with RhD(-) were 61.40%, 34.21% and 4.39% respectively, while those in the unrelated Chinese Uighurs with RhD(-) were 94.44%, 2.78% and 2.78% respectively. Furthermore, all 6 cases of some other minorities were RHD-/RHD- types. The percentage of RHD-/RHD- and RHD+/RHD- genotypes ascertained in the unrelated Chinese Uighurs were significantly higher than those in Chinese Hans (P < 0.01), whereas no statistically significant difference in the percentage of RHD+/RDH+ genotype between the two groups was observed (P > 0.05). CONCLUSION: The Rh blood group of Uighurs in Xingjiang possesses both Oriental and Caucasian characteristics, which embodies a special ethnical aspect of the Chinese nation and is in accord with the anthropologic research results. PMID- 16215958 TI - [Genetic polymorphism of Y-chromosome short tandem repeat in Elunchun ethnic group of China]. AB - OBJECTIVE: To reveal the allelic frequencies and haplotype frequencies of ten Y chromosome short tandem repeats(STR) loci (DYS392, DYS438, DYS439, DYS456, DYS459, DYS460,DYS461, DYS462, DYS389I and DYS389II systems) in an Elunchun population sample. METHODS: PCR and polyacrylamidegel electrophoresis(PAGE) followed by silver staining were performed to analyze the blood samples from 102 unrelated Elunchun male individuals. RESULTS: The allele diversity values for each locus ranged from 0.418(DYS461) to 0.727(DYS389 I). Gene diversities were higher than 0.5 except DYS462(0.479) and DYS461 (GATAA7.2)(0.418). Furthermore, 101 haplotypes from 10 Y-chromosome STRs loci were found in the sample and a high haplotype diversity 0.99 was determined. CONCLUSION: The above findings suggest that the ten Y-chromosome STR loci are valuable Y-specific markers for establishing Y-STR database, understanding human origin, paternity testing and personal identification. PMID- 16215957 TI - [Polymorphism of HLA-A,-B and DRB1 in Han population of Shanxi province]. AB - OBJECTIVE: To investigate the polymorphism of HLA-A, -B, -DRB1 genes in Han population in Shanxi of China. METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) technique was used to identify the polymorphism of HLA A, -B, -DRB1 genes of 7440 healthy and unrelated individuals of Han population in Shanxi, and the gene frequency distribution of HLA-A, -B, -DRB1 genes in population was compared with the results from other populations. RESULTS: Eighteen HLA-A, forty HLA-B and thirteen HLA-DRB1 alleles were found. The frequencies of A*02, A*24, A*11, A*01, A*03, B*13, B*51, B*15, B*40, B*35, DRB1*15, DR*09, DR*12, DR*04, DR*07 alleles in Hans of Shanxi were significantly higher and displayed distinctive distribution profiles when compared with those of Caucasian and Afro-American. CONCLUSION: The HLA-A,-B,-DRB1 distribution in Shanxi Han population shares some genetic characteristics with other Han populations in northern part of China, but it exhibits its own characteristics. PMID- 16215959 TI - The outlook for peptide drugs and the intricate relationship between the Active Pharmaceutical Ingredients manufacturer and their sponsors. AB - As research laboratories discover an ever-increasing number of peptides of pharmacological interest, there is an increased need for Good Manufacturing Practices (GMP) services, as these drugs candidates undergo clinical trials. It is therefore essential to understand the importance of the relationship with the Active Pharmaceutical Ingredients (API) manufacturer and its implications in the development and commercialization of the future peptide drug. PMID- 16215960 TI - In vivo multiple-mouse MRI at 7 Tesla. AB - We developed a live high-field multiple-mouse magnetic resonance imaging method to increase the throughput of imaging studies involving large numbers of mice. Phantom experiments were performed in 7 shielded radiofrequency (RF) coils for concurrent imaging on a 7 Tesla MRI scanner outfitted with multiple transmit and receive channels to confirm uniform signal-to-noise ratio and minimal ghost artifacts across images from the different RF coils. Grid phantoms were used to measure image distortion in different positions in the coils. The brains of 7 live mice were imaged in 3D in the RF coil array, and a second array of 16 RF coils was used to 3D image the whole bodies of 16 fixed, contrast agent-perfused mice. The images of the 7 live mouse brains at 156 microm isotropic resolution and the 16 whole fixed mice at 100 microm isotropic resolution were of high quality and free of artifacts. We have thus shown that multiple-mouse MRI increases throughput for live and fixed mouse experiments by a factor equaling the number of RF coils in the scanner. PMID- 16215961 TI - Detection of the myo-inositol 4.06-ppm resonance by selective J rewinding: application to human prefrontal cortex in vivo. AB - A new proton NMR single-voxel spectral editing strategy for the rapid measurement of myo-inositol in human brain is proposed. The spectral editing detects the 4.06 ppm, weakly coupled resonance by means of selective J rewinding. An 84.6-ms-long quadruple-resonance selective 180 degrees radiofrequency pulse, implemented within an adiabatic-refocused localization sequence, induces an in-phase triplet at 4.06 ppm, while eliminating the contribution from creatine, phosphorylethanolamine, lactate, and serine in this spectral region. The myo inositol concentration in human prefrontal cortex is estimated to be 5.7 +/- 0.9 mumol/g (mean +/- SD, n = 7), with reference to NAA at 10 micromol/g. PMID- 16215962 TI - Magnetic field shift due to mechanical vibration in functional magnetic resonance imaging. AB - Mechanical vibrations of the gradient coil system during readout in echo-planar imaging (EPI) can increase the temperature of the gradient system and alter the magnetic field distribution during functional magnetic resonance imaging (fMRI). This effect is enhanced by resonant modes of vibrations and results in apparent motion along the phase encoding direction in fMRI studies. The magnetic field drift was quantified during EPI by monitoring the resonance frequency interleaved with the EPI acquisition, and a novel method is proposed to correct the apparent motion. The knowledge on the frequency drift over time was used to correct the phase of the k-space EPI dataset. Since the resonance frequency changes very slowly over time, two measurements of the resonance frequency, immediately before and after the EPI acquisition, are sufficient to remove the field drift effects from fMRI time series. The frequency drift correction method was tested "in vivo" and compared to the standard image realignment method. The proposed method efficiently corrects spurious motion due to magnetic field drifts during fMRI. PMID- 16215963 TI - Mechanisms of tissue-iron relaxivity: nuclear magnetic resonance studies of human liver biopsy specimens. AB - MRI is becoming an increasingly important tool to assess iron overload disorders, but the complex nature of proton-iron interactions has troubled noninvasive iron quantification. Intersite and intersequence variability as well as methodological inaccuracies have been limiting factors to its widespread clinical use. It is important to understand the underlying proton relaxation mechanisms within the (human) tissue environment to address these differences. In this respect, NMR relaxometry was performed on 10 fresh human liver biopsy specimens taken from patients with transfusion-dependent anemia. T1 (1/R1) inversion recovery, T2 (1/R2) single echo, and multiecho T2 CPMG measurements were performed on a 60-MHz Bruker Minispectrometer. NMR parameters were compared to quantitative iron levels and tissue histology. Relaxivities R1 and R2 both increased linearly with hepatic iron content, with R2 being more sensitive to iron. CPMG data were well described by a chemical-exchange model and predicted effective iron center dimensions consistent with hemosiderin-filled lysosomes. Nonexponential relaxation was evident at short refocusing intervals with R2 and amplitude behavior suggestive of magnetic susceptibility-based compartmentalization rather than anatomic subdivisions. NMR relaxometry of human liver biopsy specimens yields unique insights into the mechanisms of tissue-iron relaxivity. PMID- 16215965 TI - Comparison of gated and non-gated fast multislice black-blood carotid imaging using rapid extended coverage and inflow/outflow saturation techniques. AB - PURPOSE: To comparatively analyze two fast in vivo multislice black-blood carotid artery vessel wall imaging techniques with and without cardiac gating. MATERIALS AND METHODS: Eight subjects with carotid artery atherosclerosis, and four healthy subjects were studied using two black-blood multislice techniques: rapid extended coverage double inversion recovery (REX-DIR), and inflow/outflow saturation band (IOSB) rapid acquisition with relaxation enhancement (RARE) multislice acquisitions. Quantitative, qualitative, and morphometric analyses were performed on images. RESULTS: Gating produced significantly lower values for the REX-DIR sequence with respect to signal intensity in muscle and the carotid artery wall, whereas it had no effect on flow suppression compared to non-gated images. For the IOSB sequences, gating had no significant effect on signal intensity of muscle and the carotid artery wall, but worsened flow suppression. REX-DIR and IOSB sequences were statistically different with respect to signal intensity of muscle (with REX-DIR sequences having lower values), while no statistical significance was observed for flow suppression and wall delineation. A morphologic analysis of the vessel wall and lumen comparing REX-DIR gated, IOSB gated, REX-DIR non-gated, and IOSB non-gated sequences revealed no significant differences between the acquisition techniques tested. CONCLUSION: Non-gated sequences may be used instead of gated sequences in atherosclerotic vessel wall imaging without compromising image quality. This may shorten examination time and improve patient comfort. PMID- 16215966 TI - Texture analysis for tissue discrimination on T1-weighted MR images of the knee joint in a multicenter study: Transferability of texture features and comparison of feature selection methods and classifiers. AB - PURPOSE: To investigate the reproducibility and transferability of texture features between MR centers, and to compare two feature selection methods and two classifiers. MATERIALS AND METHODS: Coronal T1-weighted MR images of the knees of 63 patients, divided into three groups, were included in the study. MR images were obtained at three different MR centers. Regions of interest (ROIs) were drawn in the bone marrow and fat tissue. Then texture analysis (TA) of the ROIs was performed, and the most discriminant features were identified using Fisher coefficients and POE+ACC (probability of classification error and average correlation coefficients). Based on these features, artificial neural network (ANN) and k-nearest-neighbor (k-NN) classifiers were used for tissue discrimination. RESULTS: Although the texture features differed among the MR centers, features from one center could be successfully used for tissue discrimination in texture data on MR images from other centers. The best results were achieved using the ANN classifier in combination with features selected by POE+ACC. CONCLUSION: The differences in texture features extracted from MR images from different centers seem to have only a small impact on the results of tissue discrimination. PMID- 16215967 TI - Implications of pulse sequence in structural imaging of trabecular bone. AB - PURPOSE: To investigate the SNR and image properties of 3D steady-state free precession (SSFP), fast large-angle spin echo (FLASE), gradient-recalled acquisition in steady state (GRASS), and spoiled GRASS (SPGR) for structural imaging of trabecular bone (TB). MATERIALS AND METHODS: SNR was examined theoretically and experimentally on phantoms, bone specimens, and in vivo. The bone volume fraction, TB thickness, and echo time (TE) dependence of the thickness were compared. The trabecula was modeled as a cylinder in simulations to examine the intra-voxel spin-dephasing in SSFP and GRASS. Images were acquired on a 1.5 T Siemens Sonata system (40 mT/m maximum gradient, 200 T/m/s peak slew rate). RESULTS: Within the hardware and safety limit constraints, SNR of FLASE was superior, followed by SSFP, GRASS, and SPGR. The trabecular thickness derived from gradient-echo images was 10-45% greater than that obtained with FLASE. Conversely, SSFP images delineated partial volume trabeculae better than FLASE. Simulations indicated that the artifactual thickening was more severe in SSFP than in GRASS, which was attributed to off-resonance effects from the induced gradients at the bone/marrow interface. CONCLUSION: FLASE had the highest SNR and was insensitive to susceptibility dephasing. Although SSFP has superior SNR compared to GRASS, off-resonance effects and duty cycle limitations may compromise its practicality in this application. Inc. PMID- 16215968 TI - Extrathoracic solitary fibrous tumor of the pelvic peritoneum with central malignant degeneration on CT and MRI. AB - We describe a 61-year-old man who presented with an extrathoracic solitary fibrous tumor (SFT) of the pelvic peritoneum with central malignant degeneration as seen on computed tomography (CT), magnetic resonance imaging (MRI), and histopathology. When a central focus of heterogeneity and variable contrast enhancement are identified within a fibrous tumor of the pelvis on CT or MRI, malignant degeneration of an extrathoracic SFT, although rare, should be considered as a diagnostic possibility. PMID- 16215970 TI - Application of 3D-MR image registration to monitor diseases around the knee joint. AB - PURPOSE: To estimate the accuracy and consistency of a method using a voxel-based MR image registration algorithm for precise monitoring of knee joint diseases. MATERIALS AND METHODS: Rigid body transformation was calculated using a normalized cross-correlation (NCC) algorithm involving simple manual segmentation of the bone region based on its anatomical features. The accuracy of registration was evaluated using four phantoms, followed by a consistency test using MR data from the 11 patients with knee joint disease. RESULTS: The registration accuracy in the phantom experiment was 0.49+/-0.19 mm (SD) for the femur and 0.56+/-0.21 mm (SD) for the tibia. The consistency value in the experiment using clinical data was 0.69+/-0.25 mm (SD) for the femur and 0.77+/-0.37 mm (SD) for the tibia. These values were all smaller than a voxel (1.25 x 1.25 x 1.5 mm). CONCLUSION: The present method based on an NCC algorithm can be used to register serial MR images of the knee joint with error on the order of a sub-voxel. This method would be useful for precisely assessing therapeutic response and monitoring knee joint diseases; normalized cross-correlation; accuracy. PMID- 16215969 TI - Multi-contrast delayed enhancement provides improved contrast between myocardial infarction and blood pool. AB - PURPOSE: To develop and test a delayed-enhancement imaging method for improving the contrast between myocardial infarction (MI) and blood pool. MATERIALS AND METHODS: The T(2) of blood is significantly longer than that of acute or chronic MI. The proposed multi-contrast delayed-enhancement (MCODE) imaging method produces a series of images with both T(1) and T(2) weightings, which provides both excellent contrast between normal and infarcted myocardium, and between blood and MI. RESULTS: The subendocardial border between MI and blood pool was easily discriminated in the T(2)-weighted image. The measured MI-to-blood contrast-to-noise ratio (CNR) was better in the T(2)-weighted image than in the T(1)-weighted image (22.5+/-8.7 vs. 2.9+/-3.1, mean+/-SD, N=11, P<0.001, for True FISP, and 19.4+/-10.8 vs. 3.9+/-2.3, N=11, P<0.001, for Turbo FLASH). CONCLUSION: The MCODE method provides a significant improvement in the ability to easily discriminate subendocardial MI by providing a T(2)-weighted image with high contrast between blood and MI. MCODE should improve both the detection and accurate sizing of MI. PMID- 16215971 TI - Planum parietale of chimpanzees and orangutans: a comparative resonance of human like planum temporale asymmetry. AB - We have previously demonstrated that leftward asymmetry of the planum temporale (PT), a brain language area, was not unique to humans since a similar condition is present in great apes. Here we report on a related area in great apes, the planum parietale (PP). PP in humans has a rightward asymmetry with no correlation to the L>R PT, which indicates functional independence. The roles of the PT in human language are well known while PP is implicated in dyslexia and communication disorders. Since posterior bifurcation of the sylvian fissure (SF) is unique to humans and great apes, we used it to determine characteristics of its posterior ascending ramus, an indicator of the PP, in chimpanzee and orangutan brains. Results showed a human-like pattern of R>L PP (P = 0.04) in chimpanzees with a nonsignificant negative correlation of L>R PT vs. R>L PP (CC = -0.3; P = 0.39). In orangutans, SF anatomy is more variable, although PP was nonsignificantly R>L in three of four brains (P = 0.17). We have now demonstrated human-like hemispheric asymmetry of a second language-related brain area in great apes. Our findings persuasively support an argument for addition of a new component to the comparative neuroanatomic complex that defines brain language or polymodal communication areas. PP strengthens the evolutionary links that living great apes may offer to better understand the origins of these progressive parts of the brain. Evidence mounts for the stable expression of a neural foundation for language in species that we recently shared a common ancestor with. PMID- 16215972 TI - Structure and growth of ultrasmall protein microcrystals by synchrotron radiation: I. microGISAXS and microdiffraction of P450scc. AB - Ultrasmall P450scc cytochrome microcrystals are grown by classical hanging vapor diffusion and by its modification using homologous protein thin-film template displaying a long-range order. The nucleation and growth mechanisms of P450scc microcrystals are studied at the thin cytochrome film surface by a new microbeam grazing incidence small angle X-ray scattering (microGISAXS) technique developed at the microfocus beamline of the European Synchrotron Radiation Facility (ESRF) in Grenoble, France. P450scc cytochrome crystals of about 5 microm are also investigated by synchrotron radiation diffraction in order to attempt a preliminary analysis of the atomic structure of this unique protein system yet unsolved. PMID- 16215973 TI - Structure and growth of ultrasmall protein microcrystals by synchrotron radiation: II. microGISAX and microscopy of lysozyme. AB - The early steps of growth and nucleation of the lysozyme microcrystals by classical and nanotemplate-based hanging vapor diffusion methods are studied using microGISAXS at the European Synchrotron Radiation Facility (ESRF) in Grenoble, France. Out-of-plane cuts in the Yoneda regions of the 2D scattering profiles point to the detection of ultrasmall lysozyme crystals by microGISAXS quite before than by light microscopy. Furthermore lysozyme crystal formation occurs quite earlier with the nanotemplate than with the classical method. Our data are compatible with two distinct modes of crystal nucleation and growth for P450sc and lysozyme. PMID- 16215974 TI - Self-assembly of nanoparticles on live bacterium: an avenue to fabricate electronic devices. PMID- 16215975 TI - Design of a mechanism-based probe for neuraminidase to capture influenza viruses. PMID- 16215976 TI - The importance of a beta-beta bond for long-range antiferromagnetic coupling in directly linked copper(II) and silver(II) diporphyrins. PMID- 16215977 TI - Bulk preparation of Si-SiOx hierarchical structures: high-density radially oriented amorphous silica nanowires on a single-crystal silicon nanocore. PMID- 16215978 TI - Spiraling steroids: organic crystals with asymmetric nanometer-scale channels. PMID- 16215979 TI - Microarray-based CGH of sporadic and syndrome-related pheochromocytomas using a 0.1-0.2 Mb bacterial artificial chromosome array spanning chromosome arm 1p. AB - Pheochromocytomas (PCC) are relatively rare neuroendocrine tumors, mainly of the adrenal medulla. They arise sporadically or occur secondary to inherited cancer syndromes, such as multiple endocrine neoplasia type II (MEN2), von Hippel-Lindau disease (VHL), or neurofibromatosis type I (NF1). Loss of 1p is the most frequently encountered genetic alteration, especially in MEN2-related and sporadic PCC. Previous studies have revealed three regions of common somatic loss on chromosome arm 1p, using chromosome-based comparative genomic hybridization (CGH) and LOH analysis. To investigate these chromosomal aberrations with a higher resolution and sensitivity, we performed microarray-based CGH with 13 sporadic and 11 syndrome-related (10 MEN2A-related and 1 NF1-related) tumors. The array consisted of 642 overlapping bacterial artificial chromosome (BAC) clones mapped to 1p11.2-p36.33. Chromosomal deletions on 1p were detected in 18 of 24 cases (75%). Among 9 tumors with partial 1p loss, the deleted region was restricted to 1cen-1p32.3 in six cases (25%), indicating a region of genetic instability. The consensus regions of deletion in this study involved 1cen 1p21.1, 1p21.3-1p31.3, and 1p34.3-1p36.33. In conclusion, these data strongly suggest that chromosome arm 1p is the site for multiple tumor suppressor genes, although the potential candidate genes CDKN2C and PTPRF/LAR are not included in these regions. PMID- 16215980 TI - Introduction of an initiator element in the mouse thymidylate synthase promoter alters S phase regulation but has no effect on promoter bidirectionality. AB - The promoter of the mouse thymidylate synthase (TS) gene lacks a TATAA box and an initiator element, is bidirectional and initiates transcription at multiple start sites across broad initiation windows upstream and downstream of the 30 nt essential promoter region. The TS promoter also plays an essential role in the post-transcription regulation of TS gene expression during the G(1)-S phase transition. The goal of this study was to determine if the addition of a TATAA box or an initiator element would have a significant effect on start-site pattern, promoter bidirectionality and S phase regulation of the TS gene. A TATAA box and/or an initiator element were inserted downstream of the TS essential promoter region, and the modified promoters were used to drive expression of indicator genes. The engineered genes were transfected into cultured mammalian cells, and the effects of the mutations were determined. Addition of the TATAA box and especially the initiator element had a significant effect on the transcription start site pattern, indicating that the elements were functional. Unexpectedly, addition of one or both of these elements had no effect on promoter bidirectionality. However, inclusion of the initiator element led to a significant reduction in S phase regulation of TS mRNA levels, indicating that changes in promoter architecture can perturb normal S phase regulation of TS gene expression. PMID- 16215981 TI - MAP kinases p38 and JNK are activated by the steroid hormone 1alpha,25(OH)2 vitamin D3 in the C2C12 muscle cell line. AB - In chick skeletal muscle cell primary cultures, we previously demonstrated that 1alpha,25(OH)2-vitamin D3 [1alpha,25(OH)2D3], the hormonally active form of vitamin D, increases the phosphorylation and activity of the extracellular signal regulated mitogen-activated protein (MAP) kinase isoforms ERK1 and ERK2, their subsequent translocation to the nucleus and involvement in DNA synthesis stimulation. In this study, we show that other members of the MAP kinase superfamily are also activated by the hormone. Using the muscle cell line C2C12 we found that 1alpha,25(OH)2D3 within 1 min phosphorylates and increases the activity of p38 MAPK. The immediately upstream mitogen-activated protein kinase kinases 3/6 (MKK3/MKK6) were also phosphorylated by the hormone suggesting their participation in p38 activation. 1Alpha,25(OH)2D3 was able to dephosphorylate/activate the ubiquitous cytosolic tyrosine kinase c-Src in C2C12 cells and studies with specific inhibitors imply that Src participates in hormone induced-p38 activation. Of relevance, 1alpha,25(OH)2D3 induced in the C2C12 line the stimulation of mitogen-activated protein kinase activating protein kinase 2 (MAPKAP-kinase 2) and subsequent phosphorylation of heat shock protein 27 (HSP27) in a p38 kinase activation-dependent manner. Treatment with the p38 inhibitor, SB203580, blocked p38 phosphorylation caused by the hormone and inhibited the phosphorylation of its downstrean substrates. 1Alpha,25(OH)2D3 also promotes the phosphorylation of c-jun N-terminal protein kinases (JNK 1/2), the response is fast (0.5-1 min) and maximal phosphorylation of the enzyme is observed at physiological doses of 1alpha,25(OH)2D3 (1 nM). The relative contribution of ERK 1/2, p38, and JNK-1/2 and their interrelationships in hormonal regulation of muscle cell proliferation and differentiation remain to be established. PMID- 16215983 TI - Evolution of sensory specializations in insectivores. AB - Although insectivores have traditionally been thought of as primitive mammals with few specializations, recent studies have revealed great diversity in the sensory systems and brain organization of members of this mammalian order. The present article reviews some of these findings in three insectivore families that are thought to form a monophyletic group. These include hedgehogs (Erinaceidae), moles (Talpidae), and shrews (Soricidae). Members of each group live in unique ecological niches, have differently specialized senses, and exhibit different behaviors. Hedgehogs have well-developed visual, auditory, and somatosensory systems. Shrews make use of visual and auditory cues, but appear to depend most heavily on touch, particularly through prominent vibrissae. Moles are somatosensory specialists with small eyes and ears and unique epidermal mechanoreceptors called Eimer's organs used to identify prey and investigate their environment. In contrast to historical views of the insectivore order, members of this group have discrete and well-organized cortical sensory areas with sharp borders as determined from both electrophysiological mapping and analysis of cortical histology. Comparison of cortical organization across species reveals a number of specializations, including expansion of cortical representations of important sensory surfaces, the addition of cortical areas to some processing networks, and the subdivision of areas into separate cortical modules. In the case of the star-nosed mole, the somatosensory system has a tactile fovea and shares a number of features in common with the visual systems of sighted mammals. PMID- 16215982 TI - Regulating leukotriene synthesis: the role of nuclear 5-lipoxygenase. AB - Leukotrienes are lipid messengers involved in autocrine and paracrine cellular signaling. They are synthesized from arachidonic acid by the 5-lipoxygenase pathway. Current models of this enzymatic pathway recognize that a key step in initiating leukotriene synthesis is the calcium-mediated movement of enzymes, including 5-lipoxygenase, to intracellular membranes. However, 5-lipoxygenase can be imported into or exported from the nucleus before calcium activation. As a result, its subcellular localization will affect its ability to be activated by calcium, as well as the membrane to which it binds and its interaction with other enzymes. This commentary focuses on the role of 5-lipoxygenase compartmentation in determining its regulation and, ultimately, leukotriene synthesis. PMID- 16215984 TI - Cellular localization of human Rad51C and regulation of ubiquitin-mediated proteolysis of Rad51. AB - Rad51-catalyzed homologous recombination is an important pathway for repair of DNA double strand breaks and maintenance of genome integrity in vertebrate cells. Five proteins referred to as Rad51 paralogs promote Rad51 activity and are proposed to act at various, and in some cases, multiple stages in the recombination pathway. Imaging studies of native Rad51 have revealed its cellular response to DNA damage, yet visualization of the paralog proteins has met with limited success. In this study, we are able to detect endogenous Rad51C and Xrcc3 in human cells. In an effort to determine how Rad51, Rad51C, and Xrcc3 influence the pattern of localization of each other over the time course of DNA damage and repair, we have made the unexpected observation that Rad51 degradation via the ubiquitin-mediated proteasome pathway occurs as a natural part of recombinational DNA repair. Additionally, we find that Rad51C plays an important role in regulating this process. This article contains supplementary material, which may be viewed at the Journal of Cellular Biochemistry website at http://www.interscience.wiley.com/jpages/0730-2312/suppmat/index.html. PMID- 16215985 TI - The RING finger protein RNF8 recruits UBC13 for lysine 63-based self polyubiquitylation. AB - The heterodimeric ubiquitin conjugating enzyme (E2) UBC13-UEV mediates polyubiquitylation through lysine 63 of ubiquitin (K63), rather than lysine 48 (K48). This modification does not target proteins for proteasome-dependent degradation. Searching for potential regulators of this variant polyubiquitylation we have identified four proteins, namely RNF8, KIA00675, KF1, and ZNRF2, that interact with UBC13 through their RING finger domains. These domains can recruit, in addition to UBC13, other E2s that mediate canonical (K48) polyubiquitylation. None of these RING finger proteins were known previously to recruit UBC13. For one of these proteins, RNF8, we show its activity as a ubiquitin ligase that elongates chains through either K48 or K63 of ubiquitin, and its nuclear co-localization with UBC13. Thus, our screening reveals new potential regulators of non-canonical polyubiquitylation. PMID- 16215986 TI - The control of chondrogenesis. AB - Chondrogenesis is the earliest phase of skeletal development, involving mesenchymal cell recruitment and migration, condensation of progenitors, and chondrocyte differentiation, and maturation and resulting in the formation of cartilage and bone during endochondral ossification. This process is controlled exquisitely by cellular interactions with the surrounding matrix, growth and differentiation factors, and other environmental factors that initiate or suppress cellular signaling pathways and transcription of specific genes in a temporal-spatial manner. Vertebrate limb development is controlled by interacting patterning systems involving prominently the fibroblast growth factor (FGF), bone morphogenetic protein (BMP), and hedgehog pathways. Both positive and negative signaling kinases and transcription factors, such as Sox9 and Runx2, and interactions among them determine whether the differentiated chondrocytes remain within cartilage elements in articular joints or undergo hypertrophic maturation prior to ossification. The latter process requires extracellular matrix remodeling and vascularization controlled by mechanisms that are not understood completely. Recent work has revealed novel roles for mediators such as GADD45beta, transcription factors of the Dlx, bHLH, leucine zipper, and AP-1 families, and the Wnt/beta-catenin pathway that interact at different stages during chondrogenesis. PMID- 16215987 TI - Tandem affinity purification revealed the hypusine-dependent binding of eukaryotic initiation factor 5A to the translating 80S ribosomal complex. AB - Eukaryotic initiation factor 5A (eIF5A) is the only protein in nature that contains hypusine, an unusual amino acid formed post-translationally in two steps by deoxyhypusine synthase and deoxyhypusine hydroxylase. Genes encoding eIF5A or deoxyhypusine synthase are essential for cell survival and proliferation. To determine the physiological function of eIF5A, we have employed the tandem affinity purification (TAP) method and mass spectrometry to search for and identify the potential eIF5A-interacting proteins. The TAP-tag was fused in-frame to chromosomal TIF51A gene and eIF5A-TAP fusion protein expressed at its natural level was used as the bait to fish out its interacting partners. At salt concentrations of 150 mM, deoxyhypusine synthase was the only protein bound to eIF5A. As salt concentrations were lowered to 125 mM or less, eIF5A interacted with a set of proteins, which were identified as the components of the 80S ribosome complex. The eIF5A-ribosome interaction was sensitive to RNase and EDTA treatments, indicating the requirement of RNA and the joining of 40S and 60S ribosomal subunits for the interaction. Importantly, a single mutation of hypusine to arginine completely abolished the eIF5A-ribosome interaction. Sucrose gradient sedimentation analysis of log versus stationary phase cells and eIF3 mutant strain showed that the endogenous eIF5A co-sedimented with the actively translating 80S ribosomes and polyribosomes in an RNase- and EDTA-sensitive manner. Our study demonstrates for the first time that eIF5A interacts in a hypusine-dependent manner with a molecular complex rather than a single protein, suggesting that the essential function of eIF5A is mostly likely mediated through its interaction with the actively translating ribosomes. PMID- 16215988 TI - Cytoprotective effect of phloroglucinol on oxidative stress induced cell damage via catalase activation. AB - We investigated the cytoprotective effect of phloroglucinol, which was isolated from Ecklonia cava (brown alga), against oxidative stress induced cell damage in Chinese hamster lung fibroblast (V79-4) cells. Phloroglucinol was found to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, hydrogen peroxide (H(2)O(2)), hydroxy radical, intracellular reactive oxygen species (ROS), and thus prevented lipid peroxidation. As a result, phloroglucinol reduced H(2)O(2) induced apoptotic cells formation in V79-4 cells. In addition, phloroglucinol inhibited cell damage induced by serum starvation and radiation through scavenging ROS. Phloroglucinol increased the catalase activity and its protein expression. In addition, catalase inhibitor abolished the protective effect of phloroglucinol from H(2)O(2) induced cell damage. Furthermore, phloroglucinol increased phosphorylation of extracellular signal regulated kinase (ERK). Taken together, the results suggest that phloroglucinol protects V79-4 cells against oxidative damage by enhancing the cellular catalase activity and modulating ERK signal pathway. PMID- 16215990 TI - Removal of kinetic traps and enhanced protein folding by strategic substitution of amino acids in a model alpha-helical hairpin peptide. AB - The presence of non-native kinetic traps in the free energy landscape of a protein may significantly lengthen the overall folding time so that the folding process becomes unreliable. We use a computational model alpha-helical hairpin peptide to calculate structural free energy landscapes and relate them to the kinetics of folding. We show how protein engineering through strategic changes in only a few amino acid residues along the primary sequence can greatly increase the speed and reliability of the folding process, as seen experimentally. These strategic substitutions also prevent the formation of long-lived misfolded configurations that can cause unwanted aggregations of peptides. These results support arguments that removal of kinetic traps, obligatory or nonobligatory, is crucial for fast folding. PMID- 16215989 TI - Knocking down PML impairs p53 signaling transduction pathway and suppresses irradiation induced apoptosis in breast carcinoma cell MCF-7. AB - The promyelocytic leukemia (PML) can selectively and dynamically recruit a number of proteins including p53 to form a sub-nuclear multiprotein chamber named PML NBs. In DNA damage response, p53 is recruited into PML-NBs and modified by phosphorylations and acetylations, which in turn potentiate its transcriptional and pro-apoptotic activities. In contrast, in carcinoma cells, the role of PML in the irradiation induced p53-mediated apoptosis is not precisely understood. In this study, we have used the breast carcinoma cell line, MCF-7, and stably suppressed the expression of PML. Inhibition of PML expression had no detectable effect on the expression of endogenous p53 at the mRNA level; however, a significant decrease of p53 protein was observed. There was also an increase in the p53-MDM2 complexes, which may facilitate p53 protein degradation by the ubiquitin-proteasome pathway, also in irradiation treated cells. The p53 transcriptional activity was attenuated both in unstressed and 10 Gy irradiation treated cells. Moreover, inhibition of PML expression in MCF-7 cells significantly reduced p53 downstream genes, cell cycle arrest gene p21(WAF/cip-1) and pro-apoptotic gene Bax expression, then irradiation-induced apoptosis. These results suggest that PML is a key regulator in the irradiation activated p53 apoptotic pathway in breast carcinoma cells. PMID- 16215991 TI - Specific anion effects on the optical rotation of glucose and serine. AB - Optical activity is directly related to molecular conformation through the anisotropic polarizabilities of molecules and the refractive index of materials. L-amino acids and D-sugars are characteristic essential bioactive molecules. Since molecular recognition and enzyme activity are related to the conformation of substrates, the relevance of optical activity to biological processes is evident. Specific ion, or Hofmeister, effects that occur with electrolytes at moderately high concentrations modify the behavior of interfaces, molecular forces between membranes, of bulk solutions, of enzymes, and even of DNA. Such effects are universal. Here we report a study on the change in optical rotation induced by some sodium salts for the enantiomers of serine and glucose in water solution. The optical rotation is shown to depend on the kind of anion and on the salt concentration. To obtain further insights into the mechanism behind the phenomenon, Fourier transform infrared (FTIR) spectral studies of serine and glucose solutions in electrolytes were also carried out. The results suggest that it is the differences in interactions of anions at specific chemical sites of the solutes that are responsible for the effects. These forces depend strongly on anion polarizability in water. Such specific ion preferential interactions can affect conformation and internal field, and result in significant changes in optical rotation. PMID- 16215992 TI - Cell differentiation dependent expressed CCR6 mediates ERK-1/2, SAPK/JNK, and Akt signaling resulting in proliferation and migration of colorectal cancer cells. AB - The expression of CCL20 (MIP-3alpha), which chemoattracts leukocytes to sites of inflammation, has been shown in intestinal epithelial cells (IEC). Aim of this study was to analyze the role of the CCL20 receptor CCR6 in IEC and colorectal cancer (CRC) cells. Expression of CCR6 and CCL20 was analyzed by RT-PCR and immunohistochemistry. Signaling was investigated by Western blotting, proliferation by MTS assays and chemotactic cell migration by wounding assays. The effect of CCL20 on Fas-induced apoptosis was determined by flow cytometry. CCR6 and its ligand CCL20 are expressed in IEC. Moreover, CRC and CRC metastases express CCR6, which is upregulated during IEC differentiation. Stimulation of IEC with CCL20 and proinflammatory stimuli (TNF-alpha, IL-1beta, LPS) significantly upregulates CCL20 mRNA expression. CCL20 expression was significantly increased in inflamed colonic lesions in Crohn's disease and correlated significantly with the IL-8 mRNA expression in these lesions (r = 0.71) but was downregulated in CRC metastases. CCL20 activated Akt, ERK-1/2, and SAPK/JNK MAP kinases and increased IL-8 protein expression. The CCL20 mediated activation of these pathways resulted in a 2.6-fold increase of cell migration (P = 0.001) and in a significant increase of cell proliferation (P < 0.05) but did not influence Fas-induced apoptosis. In conclusion, IEC and CRC express CCL20 and its receptor CCR6. CCL20 expression is increased in intestinal inflammation, while CCR6 is upregulated during cell differentiation. CCR6 mediated signals result in increased IEC migration and proliferation suggesting an important role in intestinal homeostasis and intestinal inflammation by mediating chemotaxis of IEC but also in mediating migration of CRC cells. PMID- 16215993 TI - HGF induction of postsynaptic specializations at the neuromuscular junction. AB - A critical event in the formation of vertebrate neuromuscular junctions (NMJs) is the postsynaptic clustering of acetylcholine receptors (AChRs) in muscle. AChR clustering is triggered by the activation of MuSK, a muscle-specific tyrosine kinase that is part of the functional receptor for agrin, a nerve-derived heparan sulfate proteoglycan (HSPG). At the NMJ, heparan sulfate (HS)-binding growth factors and their receptors are also localized but their involvement in postsynaptic signaling is poorly understood. In this study we found that hepatocyte growth factor (HGF), an HS-binding growth factor, surrounded muscle fibers and was localized at NMJs in rat muscle sections. In cultured Xenopus muscle cells, HGF was enriched at spontaneously occurring AChR clusters (hot spots), where HSPGs were also concentrated, and, following stimulation of muscle cells by agrin or cocultured neurons, HGF associated with newly formed AChR clusters. HGF presented locally to cultured muscle cells by latex beads induced new AChR clusters and dispersed AChR hot spots, and HGF beads also clustered phosphotyrosine, activated c-Met, and proteins of dystrophin complex; clustering of AChRs and associated proteins by HGF beads required actin polymerization. Lastly, although bath-applied HGF alone did not induce new AChR clusters, addition of HGF potentiated agrin-dependent AChR clustering in muscle. Our findings suggest that HGF promotes AChR clustering and synaptogenic signaling in muscle during NMJ development. PMID- 16215994 TI - Increased expression of deltaCaMKII isoforms in skeletal muscle regeneration: Implications in dystrophic muscle disease. AB - The expression of delta isoforms of calcium-calmodulin/dependent protein kinase II (CaMKII) has been reported in mammalian skeletal muscle; however, their functions in this tissue are largely unknown. This study was conducted to determine if deltaCaMKII expression was altered during regeneration of skeletal muscle fibers in two distinct models. In the first model, necrosis and regeneration were induced in quadriceps of normal mice by intramuscular administration of 50% glycerol. Immunostaining and confocal microscopy revealed that deltaCaMKII expression was clearly enhanced in fibers showing centralized nuclei. The second model was the mdx mouse, which undergoes enhanced muscle necrosis and regeneration due to a mutation in the dystrophin gene. sern blot analysis of hind leg extracts from 4 to 6 week old mdx mice revealed that deltaCaMKII content was decreased when compared to age-matched control mice. This loss in delta kinase content was seen in myofibrillar and membrane fractions and was in contrast to unchanged deltaCaMKII levels in cardiac and brain extracts from dystrophic mice. Confocal microscopy of mdx quadriceps and tibialis muscle showed that deltaCaMKII expression was uniformly decreased in most fibers from dystrophic mice; however, enhanced kinase expression was observed in regenerating muscle fibers. These data support a fundamental role for deltaCaMKII in the regeneration process of muscle fibers in normal and mdx skeletal muscle and may have important implications in the reparative process following muscle death. PMID- 16215995 TI - Effect of bax deletion on ethanol sensitivity in the neonatal rat cerebellum. AB - The developing cerebellum is highly sensitive to ethanol during discrete neonatal periods. This sensitivity has been linked to ethanol-induced alterations in molecules of the Bcl-2 survival-regulatory gene family. Ethanol exposure during peak periods of cerebellar sensitivity, for example, results in increased expression of proapoptotic proteins of this family, while overexpression of the antiapoptotic Bcl-2 protein in the nervous system protects against ethanol neurotoxicity. For the present study, neonatal mice with a targeted deletion of the proapoptotic bax gene were used to determine whether elimination of this protein would mitigate ethanol toxicity. bax knock-out and wild-type mice pups were exposed to ethanol via vapor inhalation during the maximal period of neonatal cerebellar ethanol sensitivity and cerebellar tissue was subsequently assessed for Purkinje and granule cell number and ethanol-mediated generation of reactive oxygen species (ROS). The results revealed that: (1) ethanol exposure during the peak period of cerebellar vulnerability resulted in substantial loss of Purkinje cells in wild-type animals, but not in bax knock-outs; (2) granule cells in the bax gene-deleted animals were not similarly protected from ethanol effects; and (3) levels of ROS following acute ethanol exposure were appreciably enhanced in the wild-type animals but not in the bax knock-outs. These results imply that Bax is important to ethanol-induced Purkinje cell death during critical neonatal periods, but that ethanol effects on granule cells may function at least partially independent of this apoptosis agonist. Amelioration of ethanol mediated increases in ROS production in the knock-outs may contribute to the observed effects. PMID- 16215996 TI - Localization of estrogen receptor-alpha and -betamRNA in brain areas controlling sexual behavior in Japanese quail. AB - Two estrogen receptors (ERs), denoted ERalpha and ERbeta, have been identified in humans and various animal species, including the Japanese quail. Estrogens play a key role in sexual differentiation and in activation of sexual behavior in Japanese quail. The distribution of ERalpha in the brain of male and female adult quail has previously been studied using immunohistochemistry, whereas in situ hybridization has been employed to study the distribution of ERbeta mRNA in males only. In this article, we used in situ hybridization to study the distribution of mRNAs for both ERalpha and ERbeta in brain areas controlling sexual behavior of Japanese quail. Our results show that both ERalpha mRNA and ERbeta mRNA are localized in areas important for sexual behavior, such as the preoptic area and associated limbic areas, in both males and females. Moreover, we found differences in distribution of mRNA for the two receptors in these areas. The results of this article support previously reported data and provide novel data on localization of ER mRNAs in adult quail brain of both sexes. PMID- 16215997 TI - R-Ras3/(M-Ras) is involved in thermal adaptation in the critical period of thermal control establishment. AB - Neuroanatomically, the body temperature is balanced by the preoptic anterior hypothalamus (PO/AH) and controlled by thermosensitive neurons. Hot or cold exposure during the critical period of temperature control development causes a plastic change in the ratio between hot- and cold-sensitive cells and can modulate temperature tolerance. In this project, mRNA fingerprinting was used to identify the proteins involved in thermal adaptation in 3-day-old chicks. Fifteen genes were induced, among which were NADH dehydrogenase, protocadherin, anolase alpha, 14-3-3epsilon, and R-Ras3. The role of each of these genes is potentially interesting and requires detailed evaluation, but since the present working hypothesis assumed neuronal remodeling, we concentrated on the role of R-Ras3/(M Ras), which is uniquely expressed in the brain and whose physiological role has not been described. In the present study, R-Ras3 expression during thermal conditioning was investigated by several molecular techniques and its mRNA was found to be induced in the PO/AH with a tenfold peak after 12 h of heat conditioning and a fourfold increase after 6 h of cold conditioning. To improve our understanding of thermal adaptation-related signal transduction, we screened for changes in the expression of transcription factors that were implicated with the Ras gene family, and found that both jun mRNA expression and Jun phosphorylation were induced after 30 min of temperature conditioning. Taken together, the present findings correlate the R-Ras3-jun pathway with thermal control establishment. PMID- 16215998 TI - Wasp venom injected into the prey's brain modulates thoracic identified monoaminergic neurons. AB - The wasp Ampulex compressa injects a cocktail of neurotoxins into the brain of its cockroach prey to induce an enduring change in the execution of locomotory behaviors. Our hypothesis is that the venom injected into the brain indirectly alters the activity of monoaminergic neurons, thus changing the levels of monoamines that tune the central synapses of locomotory circuits. The purpose of the present investigation was to establish whether the venom alters the descending control, from the brain, of octopaminergic neurons in the thorax. This question was approached by recording the activity of specific identified octopaminergic neurons after removing the input from the brain or after a wasp sting into the brain. We show that the activity of these neurons is altered in stung and "brainless" animals. The spontaneous firing rate of these neurons in stung and brainless animals is approximately 20% that in control animals. Furthermore, we show that an identified octopamine neuron responds more weakly both to sensory stimuli and to direct injection of current in all treated groups. The alteration in the activity of octopamine neurons is likely to be part of the mechanism by which the wasp induces a change in the behavioral state of its prey and also affects its metabolism by reducing the potent glycolytic activator fructose 2,6-bisphosphate in leg muscle. To our knowledge, this is the first direct evidence of a change in electrical activity of specific monoaminergic neurons that can be so closely associated with a venom-induced change in behavioral state of a prey animal. PMID- 16215999 TI - Cdc42 participates in the regulation of ADF/cofilin and retinal growth cone filopodia by brain derived neurotrophic factor. AB - Rho family GTPases have important roles in mediating the effects of guidance cues and growth factors on the motility of neuronal growth cones. We previously showed that the neurotrophin BDNF regulates filopodial dynamics on growth cones of retinal ganglion cell axons through activation of the actin regulatory proteins ADF and cofilin by inhibiting a RhoA-dependent pathway that phosphorylates (inactivates) ADF/cofilin. The GTPase Cdc42 has also been implicated in mediating the effects of positive guidance cues. In this article we investigated whether Cdc42 is involved in the effects of BDNF on filopodial dynamics. BDNF treatment increases Cdc42 activity in retinal neurons, and neuronal incorporation of constitutively active Cdc42 mimics the increases in filopodial number and length. Furthermore, constitutively active and dominant negative Cdc42 decreased and increased, respectively, the activity of RhoA in retinal growth cones, indicating crosstalk between these GTPases in retinal growth cones. Constitutively active Cdc42 mimicked the activation of ADF/cofilin that resulted from BDNF treatment, while dominant negative Cdc42 blocked the effects of BDNF on filopodia and ADF/cofilin. The inability of dominant negative Cdc42 to block ADF/cofilin activation and stimulation of filopodial dynamics by the ROCK inhibitor Y-27632 indicate interaction between Cdc42 and RhoA occurs upstream of ROCK. Our results demonstrate crosstalk occurs between GTPases in mediating the effects of BDNF on growth cone motility, and Cdc42 activity can promote actin dynamics via activation of ADF/cofilin. PMID- 16216000 TI - Correlates of breast reconstruction: results from a population-based study. AB - BACKGROUND: Immediate or early postmastectomy breast reconstruction is performed infrequently. To the authors' knowledge, little is known regarding surgeon or patient perspectives on reconstruction treatment decisions. The purpose of the current study was to identify patient attitudes and preferences associated with breast reconstruction, and whether these differed by race. METHODS: A sample of women age < or = 79 years who were diagnosed with ductal carcinoma in situ and invasive breast carcinoma between December 2001 and January 2003 was identified from the Surveillance, Epidemiology, and End Results (SEER) registries of Detroit and Los Angeles. Eligible subjects completed a questionnaire at a mean of 7 months after diagnosis. The Wald chi-square test and logistic regression were used for data analysis. RESULTS: Of the 1844 respondents, 646 underwent a mastectomy (35.0% of the total sample) and 245 of these patients received breast reconstruction (38.0%; of the mastectomy group). On multivariate analysis, younger patient age, higher educational levels, and earlier stage of disease were found to be significantly associated with breast reconstruction. Although 78.2% of women reported that breast reconstruction was discussed, only 11.2% correctly answered 3 basic knowledge questions regarding the procedure. The desire to avoid more surgery was the most common reason for not undergoing breast reconstruction. CONCLUSIONS: The results of the current study found that the majority of women were aware of breast reconstruction but choose not to undergo the procedure. Lack of knowledge and a greater perception of barriers to the procedure were more common among African-American patients and women with a lower education level, suggesting a need for improved educational strategies. PMID- 16216001 TI - Gemcitabine as frontline treatment for cutaneous T-cell lymphoma: phase II study of 32 patients. AB - BACKGROUND: Based on the activity of gemcitabine in heavily pretreated patients with cutaneous T-cell lymphoma (CTCL), the objective of the current study was to determine the role of gemcitabine in the treatment of patients with advanced, untreated CTCL. METHODS: Between June 2002 and February 2004, 32 untreated patients with mycosis fungoides (MF) (n = 26 patients); peripheral T-cell lymphoma, unspecified (PTCLU) with exclusive skin involvement (n = 5 patients); and Sezary syndrome (SS) (n = 1 patient) were enrolled in a 7-institution, Phase II trial and treated with gemcitabine. This drug was given on Days 1, 8, and 15 of a 28-day schedule at a dose of 1200 mg/m2 intravenously over 30 minutes for a total of 6 cycles. RESULTS: Of the 32 patients studied, 7 (22%) achieved a complete response (CR) and 17 (53%) achieved a partial response (PR), whereas the remaining 8 patients showed no benefit from the treatment. Five of the CRs were confirmed histologically. The CR and PR rates were found to be the same for patients with MF and PTCLU, respectively. The median duration of CR was 10 months (range, 4-22 mos). Treatment appeared to be well tolerated; hematologic toxicity was mild and no nausea/emesis or organ toxicity was noted. CONCLUSIONS: The results of the current Phase II study demonstrate the activity of gemcitabine as a single agent in untreated CTCL patients. Further studies using gemcitabine in combination, either contemporary or sequentially, with other drugs in patients with advanced stage, untreated CTCL are needed. PMID- 16216002 TI - The cost of adjuvant chemotherapy in patients with early-stage breast carcinoma. AB - BACKGROUND: In the U.S., the majority of premenopausal patients with early-stage breast carcinoma are treated with adjuvant chemotherapy. However, to the authors' knowledge, there have been few formal analyses of adjuvant chemotherapy cost performed to date, especially in premenopausal women. The objective of the current study was to evaluate the direct medical cost of adjuvant chemotherapy in women with early-stage breast carcinoma. METHODS: The attributable cost of adjuvant chemotherapy was evaluated by comparing the total cost of care for patients with breast carcinoma (cases) during the period from diagnosis to 9 months subsequent with the cost for age-matched and gender-matched control patients without breast carcinoma over an equivalent time period. The authors identified cases from a linked database of claims records from a managed care organization, and the Cancer Surveillance System registry. Controls were identified from the managed care organization from which the cases originated. Resource prices were based on reimbursements from the managed care organization. RESULTS: The attributable cost of adjuvant chemotherapy was estimated to be dollar 23,019 (95% confidence interval, dollar 19,596-dollar 26,441), based on 1239 women. The cost appeared to decrease with increasing age at diagnosis, with total costs of dollar 26,834, dollar 19,889, and dollar 17,098 for women < 50, 50 59, and > or = 60 years, respectively. Also, costs were higher for regional versus local disease (dollar 36,076 vs. dollar 12,659), for women who had a mastectomy versus breast-conserving surgery (dollar 31,075 vs. dollar 17,889), and for women who had no comorbidities versus > or = 1 comorbidity (dollar 23,606 vs. dollar 21,340). Contributors to high chemotherapy cost included use of chemotherapy agents not included in clinical guidelines during the study period, use of supportive care agents, and hospitalizations. CONCLUSIONS: The attributable cost of adjuvant chemotherapy in patients with early-stage breast carcinoma is significant. The integration of managed care claims data with clinical data from the Cancer Surveillance System registry offered a unique opportunity to derive more informative and accurate disease burden estimates in oncology. PMID- 16216003 TI - Single-fraction radiosurgery for the treatment of spinal breast metastases. AB - BACKGROUND: The spine is the most common site of bony metastases in patients with osseous breast carcinoma metastases. Spine metastases are the source of significant pain and occasionally neurologic deficit in this patient population. Conventional external beam radiotherapy lacks the precision to allow delivery of large single-fraction doses of radiation and simultaneously limit the dose to radiosensitive structures such as the spinal cord. This study evaluated the clinical efficacy of the treatment of spinal breast carcinoma metastases with a single-fraction radiosurgical technique. METHODS: In this prospective cohort evaluation, 68 breast carcinoma metastases to the spine in 50 patients were treated with a single-fraction radiosurgery technique with a follow-up period of 6-48 months, median 16 months. The most common indication for radiosurgery treatment was pain in 57 lesions, as a primary treatment modality in 8 patients, and for radiographic tumor progression, as a postsurgical boost, and for a progressive neurologic deficit in 1 patient each. RESULTS: Tumor volume ranged from 0.8-197 cm3 (mean, 27.7 cm3). Maximum tumor dose was maintained at 15-22.5 Gy (mean, 19 Gy). No radiation-induced toxicity occurred during the follow-up period (6-48 mo). Long-term axial and radicular pain improvement occurred in 55 of 57 (96%) patients who were treated primarily for pain. Long-term radiographic tumor control was seen in all patients who underwent radiosurgery as their primary treatment modality, for radiographic tumor progression, or as a postsurgical treatment. CONCLUSIONS: Spinal radiosurgery was found to be feasible, safe, and clinically effective for the treatment of spinal metastases from breast carcinoma. The results indicate the potential of radiosurgery in the treatment of patients with spinal breast metastases, especially those with solitary sites of spine involvement, to improve long-term palliation. PMID- 16216004 TI - Plasma cytokine and P-selectin levels in advanced malignancy: prognostic value and impact of low-molecular weight heparin administration. AB - BACKGROUND: The survival benefit described in patients with cancer treated with low molecular weight heparin (LMWH) may result from a LMWH-mediated effect on the immune system or on the cross-talk between platelets and tumor cells. METHODS: Plasma levels of interleukin (IL)-10, IL-6, interferon (IFN)-gamma, and P selectin were measured in patients with advanced stage malignancy who were randomized to receive standard cancer care with or without the addition of LMWH. RESULTS: Patients with IL-6 levels above the median had a median survival of 6.5 months versus 8.8 months for those with values below this cutoff (P = 0.02). IL 10 levels were found to be similarly correlated with survival such that IL-10 concentrations above the detection limit of the assay were associated with a doubled risk of dying in comparison to undetectable IL-10 (P = 0.02). No significant association was found between survival and circulating levels of IFN gamma. For P-selectin, patients with values below the fourth quartile had a median survival of 8.8 months versus 6.5 months for patients with levels above the fourth quartile (P = 0.02). In multivariate analysis, IL-10 remained an independent unfavorable prognostic factor (hazard ratio, 2.13; 95% confidence interval, 1.08-4.20). In patients treated with LMWH, the plasma levels of IL-6, IL-10, IFN-gamma, and P-selectin demonstrated similar correlations with survival. However, none of the markers was associated with the beneficial survival effects observed with the administration of LMWH. CONCLUSIONS: IL-10, IL-6, and P selectin levels predicted a poor outcome in patients with advanced stage malignancy. The prolongation in survival observed with LMWH in patients with cancer apparently cannot be explained by a LMWH effect on these circulating markers. PMID- 16216005 TI - Hippocampal morphology is differentially affected by reproductive experience in the mother. AB - Pregnancy and mothering result in a number of hormonal, neurological, and behavioral changes that are necessary to ensure reproductive success. With subsequent reproductive experience (multiparity and mothering), further neurological and behavioral changes may result. Recent research has shown that previous motherhood enhances both hippocampus-dependent learning and memory and long-term potentiation (LTP); together with decreases in hippocampus volumes during pregnancy it is suggested that the hippocampus is affected by pregnancy and/or mothering. The present experiment aimed to investigate the effect of reproductive experience (nulli, primi-, and multiparity and mothering) on dendritic morphology in the CA1 and CA3 regions of the hippocampus. Brains were stained with a modified version of the single-section Golgi impregnation technique, and dendritic length, number of branch points, and spine density was analyzed for apical and basal regions of CA1 and CA3 pyramidal neurons. Primiparity and/or mothering resulted in dendritic remodeling in both the CA1 and CA3 hippocampal regions, and multiparity resulted in enhanced spine density in the basal CA1 region, which was positively correlated with number of male pups in a litter. These findings point to the effect of reproductive experience and offspring on plasticity in the hippocampus, an area not traditionally associated with motherhood. PMID- 16216006 TI - Tissue inhibitor of metalloproteinase-2(TIMP-2)-deficient mice display motor deficits. AB - The degradation of the extracellular matrix is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Matrix components of the basement membrane play critical roles in the development and maintenance of the neuromuscular junction (NMJ), yet almost nothing is known about the regulation of MMP and TIMP expression in either the pre- or postsynaptic compartments. Here, we demonstrate that TIMP-2 is expressed by both spinal motor neurons and skeletal muscle. To determine whether motor function is altered in the absence of TIMP-2, motor behavior was assessed using a battery of tests (e.g., RotaRod, balance beam, hindlimb extension, grip strength, loaded grid, and gait analysis). TIMP-2(-/-) mice fall off the RotaRod significantly faster than wild-type littermates. In addition, hindlimb extension is reduced and gait is both splayed and lengthened in TIMP-2(-/-) mice. Motor dysfunction is more pronounced during early postnatal development. A preliminary analysis revealed NMJ alterations in TIMP-2(-/-) mice. Juvenile TIMP-2(-/-) mice have increased nerve branching and acetylcholine receptor expression. Adult TIMP-2(-/ ) endplates are enlarged and more complex. This suggests a role for TIMP-2 in NMJ sculpting during development. In contrast to the increased NMJ nerve branching, cerebellar Purkinje cells have decreased neurite outgrowth. Thus, the TIMP-2(-/-) motor phenotype is likely due to both peripheral and central defects. The tissue specificity of the nerve branching phenotype suggests the involvement of different MMPs and/or extracellular matrix molecules underlying the TIMP-2(-/-) motor phenotype. PMID- 16216008 TI - Calmodulin is a critical regulator of osteoclastic differentiation, function, and survival. AB - Increased osteoclastic resorption and subsequent bone loss are common features of many debilitating diseases including osteoporosis, bone metastases, Paget's disease, and rheumatoid arthritis. While rapid progress has been made in elucidating the signaling pathways directing osteoclast differentiation and function, a comprehensive picture is far from complete. Here, we explore the role of the Ca(2+)-activated regulator calmodulin in osteoclastic differentiation, functional bone resorption, and apoptosis. During active bone resorption, calmodulin expression is increased, and calmodulin concentrates at the ruffled border, the organelle utilized for acid transport and bone dissolution. Pharmacologic inhibitors of calmodulin, several of which are already used clinically as anti-cancer and anti-psychotic agents, inhibit osteoclastic acid transport, suggesting their potential as bone-sparing drugs. Recent studies also implicate calmodulin in osteoclast apoptosis through a mechanism involving its direct interaction with the death receptor Fas. During osteoclastogenesis, RANKL induction stimulates a rise in intracellular Ca2+, which in turn activates calmodulin and its downstream effectors. In particular, the Ca(2+)/calmodulin dependent phosphatase calcineurin and its targets, the NFAT family of transcription factors, have been posited as the master regulators of osteoclastogenesis. However, recent in vivo and in vitro studies demonstrate that another Ca(2+)/calmodulin-regulated effector protein, CaMKII, is also involved. CaMKII(+/-) mutant mice have reduced osteoclast numbers, and CaMKII antagonists inhibit osteoclastogenesis in vitro. Furthermore, CaMKII is known to activate AP 1 transcription factors, which are also required for RANKL-induced osteoclast gene transcription, and recent findings suggest that CaMKII can down-regulate gp130, a cytokine receptor involved in bone remodeling and implicated in numerous osteo-articular diseases. PMID- 16216007 TI - Apoptosis evasion: the role of survival pathways in prostate cancer progression and therapeutic resistance. AB - The ability of a tumor cell population to grow exponentially represents an imbalance between cellular proliferation and cellular attrition. There is an overwhelming body of evidence suggesting the ability of tumor cells to avoid programmed cellular attrition, or apoptosis, is a major molecular force driving the progression of human tumors. Apoptotic evasion represents one of the true hallmarks of cancer and appears to be a vital component in the immunogenic, chemotherapeutic, and radiotherapeutic resistance that characterizes the most aggressive of human cancers [Hanahan and Weinberg, 2000]. The challenges in the development of effective treatment modalities for advanced prostate cancer represent a classic paradigm of the functional significance of anti-apoptotic pathways in the development of therapeutic resistance. PMID- 16216009 TI - Brain of the African elephant (Loxodonta africana): neuroanatomy from magnetic resonance images. AB - We acquired magnetic resonance images of the brain of an adult African elephant, Loxodonta africana, in the axial and parasagittal planes and produced anatomically labeled images. We quantified the volume of the whole brain (3,886.7 cm3) and of the neocortical and cerebellar gray and white matter. The white matter-to-gray matter ratio in the elephant neocortex and cerebellum is in keeping with that expected for a brain of this size. The ratio of neocortical gray matter volume to corpus callosum cross-sectional area is similar in the elephant and human brains (108 and 93.7, respectively), emphasizing the difference between terrestrial mammals and cetaceans, which have a very small corpus callosum relative to the volume of neocortical gray matter (ratio of 181 287 in our sample). Finally, the elephant has an unusually large and convoluted hippocampus compared to primates and especially to cetaceans. This may be related to the extremely long social and chemical memory of elephants. PMID- 16216010 TI - Aminopeptidase-N/CD13 (EC 3.4.11.2) inhibitors: chemistry, biological evaluations, and therapeutic prospects. AB - Aminopeptidase N (APN)/CD13 (EC 3.4.11.2) is a transmembrane protease present in a wide variety of human tissues and cell types (endothelial, epithelial, fibroblast, leukocyte). APN/CD13 expression is dysregulated in inflammatory diseases and in cancers (solid and hematologic tumors). APN/CD13 serves as a receptor for coronaviruses. Natural and synthetic inhibitors of APN activity have been characterized. These inhibitors have revealed that APN is able to modulate bioactive peptide responses (pain management, vasopressin release) and to influence immune functions and major biological events (cell proliferation, secretion, invasion, angiogenesis). Therefore, inhibition of APN/CD13 may lead to the development of anti-cancer and anti-inflammatory drugs. This review provides an update on the biological and pharmacological profiles of known natural and synthetic APN inhibitors. Current status on their potential use as therapeutic agents is discussed with regard to toxicity and specificity. PMID- 16216011 TI - Adaptive response of human tendon to paralysis. AB - To gain insight into the adaptive response of human tendon to paralysis, we compared the mechanical properties of the in vivo patellar tendon in six men who were spinal cord-injured (SCI) and eight age-matched, able-bodied men. Measurements were taken by combining dynamometry, electrical stimulation, and ultrasonography. Tendon stiffness and Young's modulus, calculated from force elongation and stress-strain curves, respectively, were lower by 77% (P < 0.01) and 59% (P < 0.05) in the SCI than able-bodied subjects. The cross-sectional area (CSA) of the tendon was 17% smaller (P < 0.05) in the SCI subjects, but there was no difference in tendon length between the two groups. Our results indicate that paralysis causes substantial deterioration of the structural and material properties of tendon. This needs to be taken into consideration in the design of electrical stimulation protocols for rehabilitation and experimental purposes, and when interpreting changes in the contractile speed of paralyzed muscle. PMID- 16216012 TI - Genome scans with gene-covariate interaction. AB - Genetic models for gene-covariate interactions are described. Methods of linkage analysis that utilize special features of these models and the corresponding score statistics are derived. Their power is compared with that of simple genome scans that ignore these special features, and substantial gains in power are observed when the gene-covariate interaction is strong. Quantitative trait mapping in randomly ascertained sibships and affected sibpair mapping are discussed. For the latter case, a simpler statistic is proposed that has similar performance to the score statistic, but does not require the estimation of nuisance parameters. Since the nuisance parameters are not estimable solely from affected sib-pair data, this statistic would be much easier to apply in practice. Similarities with linkage analysis of models for longitudinal data and multivariate phenotypes are also briefly discussed. Approximations for the P value and power are derived under the framework of local alternatives. PMID- 16216015 TI - Is it safe to perform cardiac catheterizations on adults with congenital heart disease in a pediatric catheterization laboratory? AB - OBJECTIVE: To determine the complication rate during the catheterization in adults with congenital heart disease (CHD) in a pediatric catheterization laboratory (PCL). BACKGROUND: An increasing number of patients with CHD are surviving into adulthood, with diagnostic and interventional cardiac catheterization being essential for the management of their disease. The complication rate during the catheterization of adults with CHD has not been reported. METHODS: A retrospective chart review was performed on all adult patients (>18 years) with CHD who underwent diagnostic or interventional catheterization in our PCL within the past 8.5 years. RESULTS: A total of 576 procedures were performed on 436 adult patients (median age 26 years). Complex heart disease was present in 387/576 (67%) procedures. An isolated atrial septal defect or patent foramen ovale was present in 115/576 (20%) procedures, and 51/576 (9%) procedures were performed on patients with structurally normal hearts with arrhythmias. Interventional catheterization was performed in 378/576 (66%) procedures. There were complications during 61/576 (10.6%) procedures; 19 were considered major and 42 minor. Major complications were death (1), ventricular fibrillation (1), hypotension requiring inotropes (7), atrial flutter (3), retroperitoneal hematoma, pneumothorax, hemothorax, aortic dissection, renal failure, myocardial ischemia and stent malposition (1 each). The most common minor complications were vascular entry site hematomas and hypotension not requiring inotropes. Procedures performed on patients > or = 45 years of age had a 19% occurrence of complications overall compared with 9% occurrence rate in patients of age < 45 years (P < 0.01). CONCLUSIONS: The complication rate during the catheterization of adults with CHD in a PCL is similar to the complication rate of children with CHD undergoing cardiac catheterization. The older subset of patients are more likely to encounter complications overall. The encountered complications could be handled effectively in the PCL. With screening in place, it is safe to perform cardiac catheterization on most adults with CHD in a PCL. PMID- 16216016 TI - Critical dose of lead affecting postural balance in workers. AB - BACKGROUND: The threshold blood lead (BPb) level for nervous system dysfunction in adults has been estimated to be 30-40 microg/dl. This study was carried out to estimate the critical dose of lead affecting the neuromotor function in workers by introducing the benchmark dose (BMD) approach, as well as to identify the specific postural sway to lead exposure. METHODS: Postural sway parameters with spectral analysis were compared between 121 lead workers with BPb levels of 6-89 (mean 40) microg/dl and 60 unexposed controls. RESULTS: All sway parameters, except for sagittal sways with eyes open, were significantly larger in the lead workers than in the controls; also, the Romberg quotient for sagittal sway was significantly higher in the lead workers. The BPb level in the lead workers was significantly related to sagittal sways at 1-2 Hz and 2-4 Hz with eyes open, and sagittal and transversal sways at 1-2 Hz and 2-4 Hz with eyes closed. The BMD levels of BPb (i.e., lower 95% confidence limits of BMD) were estimated to be 12.1-17.3 (mean 14.4) microg/dl for postural sway. CONCLUSIONS: Neuromotor dysfunction in lead workers may be initiated at BPbs below the level previously accepted as effectless, and be characterized mainly by an increased sway of high frequency (1-4 Hz) in the sagittal direction with eyes closed (high Romberg quotient). PMID- 16216018 TI - Improved survival for stenting vs. balloon angioplasty for the treatment of coronary artery disease in patients with ischemic left ventricular dysfunction. AB - While earlier studies of balloon angioplasty (BA) in patients with left ventricular (LV) dysfunction suggested high late mortality, a study directly comparing coronary stenting and BA has not been performed. Since stenting provides a more durable revascularization, we sought to compare long-term survival in patients undergoing stenting vs BA in patients with decreased left ventricular ejection fractions (LVEF). We evaluated consecutive patient procedures performed in our institution from 1996 through 1999. Patients were considered part of the stent group if they received at least one stent. To be included, patients had to have a technically adequate angiographic LV gram with a calculated LVEF0.05); five days later the inhibitory rates remained as 33.87% and 36.86% respectively (P>0.05). The pain responses of both hindlimbs in the rats with the right DC destroyed showed no significant difference compared with the intact rats. CONCLUSION: DC is not involved in the inhibitory effect of DPN on the pressor response induced by PVN stimulation. PMID- 16216057 TI - [Cardioprotection of mitoSlo1 channel activation involves mitochondrial permeability transition in ischemia and reperfusion of rat hearts]. AB - OBJECTIVE: To investigate whether the cardioprotection of mitochondrial Slo channel (mitoSlo(1) channel) is associated with mitochondrial permeability transition in isolated rat hearts subjected to ischemia and reperfusion. METHODS: Isolated perfused rat hearts were subjected to 30 min regional ischemia (occlusion of left anterior descending artery) and 120 min reperfusion. The infarct size, lactate dehydrogenase (LDH) release during reperfusion and ventricular hemodynamic parameters were measured. RESULTS: Pretreatment with mitoSlo(1) channel opener, NS1619 10 micromol/L for 10 min reduced the infarct size and LDH release, and improved the recovery of left ventricular developed pressure, left ventricular end-diastolic pressure, maximal rise/fall rate of left ventricular pressure and coronary flow during reperfusion. Administration of atractyloside (20 micromol/L), an opener of mitochondrial permeability transition pore, for 20 min (last 5 min of ischemia and first 15 min of reperfusion) attenuated the reduction of infarct size and LDH release and improvement of left ventricular performance induced by NS1619. In the isolated mitochondria, a significant inhibition of Ca(2+)-induced swelling was observed when mitochondria were incubated with NS1619. CONCLUSION: MitoSlo(1) channel activation by NS1619 protects the myocardium against ischemia and reperfusion injury by inhibiting mitochondrial permeability transition pore opening. PMID- 16216058 TI - [Influence of fluvastatin on left ventricular remodeling after myocardial infarction in rats]. AB - OBJECTIVE: To investigate the effect of long-term administration of fluvastatin on improvement of ventricular remodeling of rats after myocardial infarction and its mechanism. METHODS: Sprague-Dawley rats were subjected to ligation in anterior descending branch of coronary artery and treated with fluvastatin (20 mg.kg(-1) d(-1)) or distilled water for 8 weeks. Doppler echocardiography, hemodynamic study and cardiac histomorphometry were used to estimate the ventricular remodeling and cardiac function. Laser scanning confocal microscope was used to definite the distribution of superoxide anion (O(2)(*-)) and nitrogen monoxide. RT-PCR and immunohistochemistry were used to detect the expression of NOS2 and p22phox in mRNA and protein level. The level of lipid peroxidation, glutathione peroxidase, nitrogen monoxide and total cholesterol were detected too. RESULTS: Administration of fluvastatin ameliorated left ventricular remodeling without affecting the infarct size [(40 +/- 6 vs 42 +/-5)%, P>0.05]. The level of left ventricular end-diastolic pressure [(18.24 +/-6.58 vs 10.74 +/ 4.71) mmHg, P<0.05], right ventricular ameliorated relative weight [(0.92 +/-0.19 vs 0.71 +/-0.13) g/kg, P<0.05], the thickness of left ventricular posterior wall [(3.04 +/-0.28 vs 2.60 +/-0.36) mm, P<0.05] decreased after fluvastatin treatment. The left ventricular ejection fraction was not influenced, the relative lung weight and the left atrium diameter reduced [(5.79 +/-2.92 vs 3.69 +/-0.68) g/kg, (0.55 +/-0.12 vs 0.45 +/-0.04) mm, P<0.05]; the expressions of LPO in the plasma and myocardium [(8.64 +/-0.59 vs 7.71 +/-0.66) U/dl, P<0.05; (3.12 +/-0.38 vs 1.93 +/-0.40) ng/microg.pro, P<0.01] were reduced, and the overexpressed NO was inhibited [(436.87 +/-47.22 vs 313.78 +/-34.35) mg/dl, P<0.01], but the expression of GPx increased [(66.13 +/-8.31 vs 79.78 +/-2.38) mg/dl, P<0.01]. The expression of O(2)(*-) and the activity of NADPH oxidase subunit p22phox increased; NOS2 and its products NO were over-expressed too. CONCLUSION: Ventricular remodeling and hemodynamics are improved profoundly in MI rats treated with fluvastatin. The effect of antioxidative stress of fluvastatin might be involved in the mechanism. PMID- 16216059 TI - [Alterations of myocardial ultrastructure and gene expression of calcium handling proteins in diabetic rat heart]. AB - OBJECTIVE: To investigate the ultrastructure of myocardium and gene expression of calcium handling proteins in diabetic rat heart. METHODS: Diabetes was induced in male Sprague-Dawley rats by a single injection of alloxanm (40 mg/kg ) and the rats in control group were injected with normal saline. At the end of 2, 4, 6 weeks after the induction of diabetes, the animals were sacrificed. The expression of calcium handling proteins was detected by reverse transcription polymerase chain reaction (RT-PCR) and actin mRNA was used as internal standard. Heart tissue at the apex was obtained for light and electron microscope study. RESULTS: At the end of 4 and 6 weeks, cardiosomatic ratio of diabetic rats was higher than that of control. Electron microscopy revealed a spectrum of subcellular remodeling in myocardium which was characterized by damaged myofibrils and mitochondria, dilated and swollen sarcoplasmic reticulum. Expression of phospholamban mRNAs was significantly increased, but 1,4,5 trisphosphate inositol receptor type 2, ryanodine receptor type 2 mRNAs were significantly decreased compared with those in the age-matched control rats. In contrast, the expression of sarco/endoplasmic reticulum Ca(2+)-ATPase mRNAs was not affected. CONCLUSION: In diabetic rat heart, gene expression of calcium handling proteins was characterized by up-regulation of phospholamban and down regulation of sarcoplasmic reticulum calcium release channel while electron microscopic analysis of myocardium revealed a spectrum of subcellular remodeling. PMID- 16216060 TI - [Efficacy and safety of azosemide in patients with edema and ascites]. AB - OBJECTIVE: To assess the efficacy and safety of azosemide in patients with edema and ascites. METHODS: A multicentral, randomized, double-blind, controlled clinical trial was applied. All 223 patients (cardiac edema 92, hepatogenic edema 63, renal edema 68) were randomized to azoesmide and furosemide group, and all patients were treated for 2 weeks. Patients with cardiac or renal edema took azosemide (30 mg/d) or furosemide (20 mg/d); patients with hepatogenic edema took azosemide (60 mg/d) or furosemide (40 mg/d). The dosage were adjusted to azosemide 60 mg/d (cardiac, renal edema), 90 mg (hepatogeic edema); or furosemide 40 mg/d (cardiac, renal edema), 60 mg (hepatogeic edema), if diuretic effects were not obtained at the end of third day. RESULTS: At the end of the study, the weight changes were (2.87+/-3.10) kg and (2.81 +/-2.84) kg; the total effective rate of edema lessen was 89.19% and 89.81%; the total effective rate of heart function improvement was 64.44% and 66.66%; the 24 h urine output increased (321.85 +/-669.52) ml and (273.80 +/-645.72) ml for azosemide and furosemide, respectively. The total effective rate of ascites lessen (tested by B-ultrasound) was 89.28% and 86.66%; abdominal girth decreased (5.20 +/-3.58) cm and (5.03 +/ 3.74) cm for azosemide and furosemide, respectively. The adverse event rate was 23.01% in azosemide group and 21.01% in furosemide group; the main adverse effects were hypokalemia, hyperuricemia, hypertriglyceridemia and thirsty. CONCLUSION: Azosemide could effectively lessen edema, improve heart function and decrease ascitesuit is well tolerated and is particularly useful for the diuretic treatment. PMID- 16216061 TI - [Total thyroidectomy with a modified Miccoli's approach for treatment of Graves' disease--feasibility and its applying techniques]. AB - OBJECTIVE: To assess the feasibility and relevant applying techniques of total thyroidectomy for Graves' disease with a modified Miccoli's approach. METHODS: Forty-two patients with Graves' disease consecutively received the radical operation from June 2002 to December 2004.The modification includes: (1) Incision extending according to the degree of lobe enlargement (3-6 cm, average 4 cm); (2) A space maintain-regulating device was used to change dimensionally the volume of working space (mainly height) when specific manipulation needed; (3) A volume reducing resection step was performed for the gland with degree III hyperplasia by cutting off the middle-inferior part of the lobe prior to endoscopic lobectomy. The approach was designed to mainly use ultrasonically-activated scalpels, with suction-dissector or others as supplementary instruments. During the operations, a method of "sequenced dissect-coagulate-cut" was employed to directly divide all branches of thyroid vessels without ligation or application of hemoclips. RESULTS: All procedures were completed successfully. None of them were converted to open surgery due to uncontrolled bleeding or severe postoperative hematoma. No severe complications occurred, except 2 cases who suffered from temporary hoarseness. CONCLUSION: Total thyroidectomy for Graves' disease can be safely performed with the modified Miccoli's approach by using ultrasonic scalpel and the space maintain-regulating device. Application of these adaptive reforms can obviously reduce the difficulties in manipulation, and thus, make the usage of this minimally invasive design also clinically possible for even radical treatment of the gland. PMID- 16216062 TI - [Adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD mutation]. AB - OBJECTIVE: To evaluate the efficacy and safety of adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine therapy. METHODS: The disease relapsed in 14 hepatitis B patients with decompensated liver cirrhosis during lamivudine treatment due to the YMDD motif mutation. All 14 patients had positive HVBDNA and active hepatitis, and were evaluated as Child-Pugh Score C (CPS-C). The patients were treated with lamivudine 50 mg/d and adefovir dipivoxil 10 mg/d for 6 months. RESULTS: One patient signed off due to non-hypoxemic hyperlactacidemia; other 13 patients showed decreased serum HBVDNA. All patients had serum HBVDNA < or =10(5) copies/ml and 7 patients had HBVDNA < or =10(4) copies/ml. Six patients regained normal serum ALT level and Child-Pugh scores decreased in all patients. CONCLUSION: Adefovir dipivoxil has satisfied efficacy and safety in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine treatment. PMID- 16216063 TI - [Ultrasonic evaluation of nut cracker syndrome before and after stent implantation]. PMID- 16216064 TI - [Progress in research on two-pore-domain potassium channel family]. PMID- 16216065 TI - [Total aortic arch replacement]. PMID- 16216067 TI - Mimicking the first step of RNA splicing: an artificial DNA enzyme can synthesize branched RNA using an oligonucleotide leaving group as a 5'-exon analogue. AB - The 7S11 deoxyribozyme synthesizes 2',5'-branched RNA by mediating the nucleophilic attack of an internal 2'-hydroxyl group of one RNA substrate into the 5'-triphosphate of a second RNA substrate, with pyrophosphate as the leaving group. Here we comprehensively examined the role of the leaving group in the 7S11 catalyzed reaction by altering the 5'-phosphorylation state and the length of the second RNA substrate. When the leaving group is the less stabilized phosphate or hydroxide anion as provided by a 5'-diphosphate or 5'-monophosphate, the same 2',5'-branched product is formed as when pyrophosphate is the leaving group, but with an approximately 50- or approximately 1000-fold lower rate (Bronsted beta(LG) = -0.40). When the 5'-end of the RNA substrate that bears the leaving group is longer by one or more nucleotides, either the new 5'-terminal alpha phosphate or the original alpha-phosphate can be attacked by the branch-site 2' hydroxyl group; in the latter case, the leaving group is an oligonucleotide. The choice between these alpha-phosphate reaction sites is determined by the subtle balance between the length of the single-stranded 5'-extension and the stability of the leaving group. Because the branch-site adenosine is a bulged nucleotide flanked by Watson-Crick duplex regions, we earlier concluded that 7S11 structurally mimics the first step of natural RNA splicing. The observation of 7S11-catalyzed branch formation with an oligonucleotide leaving group strengthens this resemblance to natural RNA splicing, with the oligonucleotide playing the role of the 5'-exon in the first step. These findings reinforce the notion that splicing-related catalysis can be achieved by artificial nucleic acid enzymes that are much smaller than the spliceosome and group II introns. PMID- 16216066 TI - Molecular dynamics simulations of salicylate effects on the micro- and mesoscopic properties of a dipalmitoylphosphatidylcholine bilayer. AB - Salicylate, an amphiphilic molecule and a popular member of the nonsteroidal anti inflammatory drug family, is known to affect hearing through reduction of the electromechanical coupling in the outer hair cells of the ear. This reduction of electromotility by salicylate has been widely studied, but the molecular mechanism of the phenomenon is still unknown. In this study, we investigated one aspect of salicylate's action, namely the perturbation of electrical and mechanical membrane properties by salicylate in the absence of cytoskeletal or membrane-bound motor proteins such as prestin. In particular, we simulated the interaction of salicylate with a dipalmitoylphosphatidylcholine (DPPC) bilayer via atomically detailed molecular dynamics simulations to observe the effect of salicylate on the microscopic and mesoscopic properties of the bilayer. The results demonstrate that salicylate interacts with the bilayer by associating at the water-DPPC interface in a nearly perpendicular orientation and penetrating more deeply into the bilayer than either sodium or chloride. This association has several affects on the membrane properties. First, binding of salicylate to the membrane displaces chloride from the bilayer-water interface. Second, salicylate influences the electrostatic potential and dielectric properties of the bilayer, with significant changes at the water-lipid bilayer interface. Third, salicylate association results in structural changes, including decreased headgroup area per lipid and increased lipid tail order. However, salicylate does not significantly alter the mechanical properties of the DPPC bilayer; bulk compressibility, area compressibility, and bending modulus were only perturbed by small, statistically insignificant amounts by the presence of salicylate. The observations from these simulations are in qualitative agreement with experimental data and support the conclusion that salicylate influences the electrical but not the mechanical properties of DPPC membranes. PMID- 16216068 TI - Photolabeling the Torpedo nicotinic acetylcholine receptor with 4-azido-2,3,5,6 tetrafluorobenzoylcholine, a partial agonist. AB - The interactions of a photoreactive analogue of benzoylcholine, 4-azido-2,3,5,6 tetrafluorobenzoylcholine (APFBzcholine), with nicotinic acetylcholine receptors (nAChRs) were studied using electrophysiology and photolabeling. APFBzcholine acted as a low-efficacy partial agonist, eliciting maximal responses that were 0.3 and 0.1% of that of acetylcholine for embryonic mouse and Torpedo nAChRs expressed in Xenopus oocytes, respectively. Equilibrium binding studies of [3H]APFBzcholine with nAChR-rich membranes from Torpedo electric organ revealed equal affinities (K(eq) = 12 microM) for the two agonist binding sites. Upon UV irradiation at 254 nm, [3H]APFBzcholine was photoincorporated into the nAChR alpha, gamma, and delta subunits in an agonist-inhibitable manner. Photolabeled amino acids in the agonist binding sites were identified by Edman degradation of isolated, labeled subunit fragments. [3H]APFBzcholine photolabeled gammaLeu 109/deltaLeu-111, gammaTyr-111, and gammaTyr-117 in binding site segment E as well as alphaTyr-198 in alpha subunit binding site segment C. The observed pattern of photolabeling is examined in relation to the predicted orientation of the azide when APFBzcholine is docked in the agonist binding site of a homology model of the nAChR extracellular domain based upon the structure of the snail acetylcholine binding protein. PMID- 16216069 TI - Chemoenzymatic synthesis of cryptophycin/arenastatin natural products. AB - Microbially derived modular polyketide synthase and nonribosomal peptide synthetase biosynthetic pathways are a rich source of novel natural products. Development of these systems for the engineered biosynthesis of diverse secondary metabolites continues to progress as a robust source of chemical diversity. Recent efforts that employ individual enzymes and catalytic domains for the production or modification of small molecules have met with growing success. In this study, the thioesterase domain from the cryptophycin biosynthetic pathway was isolated and its function evaluated with a series of linear chain elongation intermediates in developing a novel chemoenzymatic synthesis of the cryptophycin/arenastatin class of antitumor agents. The results show the high efficiency of the thioesterase in generating the 16-membered depsipeptide ring of this important natural product system. Moreover, analysis of selected substrates revealed considerable tolerance for structural variation within the seco cryptophycin unit C beta-alanine residue, but strict structural requirements at the phenyl group position of the unit A delta-hydroxy octadienoate chain elongation intermediates. PMID- 16216070 TI - Mutagenesis of Glycine 179 modulates both catalytic efficiency and reduced pyridine nucleotide specificity in cytochrome b5 reductase. AB - Cytochrome b5 reductase (cb5r), a member of the ferredoxin:NADP+ reductase family of flavoprotein transhydrogenases, catalyzes the NADH-dependent reduction of cytochrome b5. Within this family, a conserved "GxGxxP" sequence motif has been implicated in binding reduced pyridine nucleotides. However, Glycine 179, a conserved residue in cb5r primary structures, precedes this six-residue "180GxGxxP185" motif that has been identified as binding the adenosine moiety of NADH. To investigate the role of G179 in NADH complex formation and NAD(P)H specificity, a series of rat cb5r variants were generated, corresponding to G179A, G179P, G179T, and G179V, recombinantly expressed in Escherichia coli and purified to homogeneity. Each mutant protein was found to incorporate FAD in a 1:1 cofactor/protein stoichiometry and exhibited absorption and CD spectra that were identical to those of wild-type cb5r, indicating both correct protein folding and similar flavin environments, while oxidation-reduction potentials for the FAD/FADH2 couple (n = 2) were also comparable to the wild-type protein (E(o)' = -272 mV). All four mutants showed decreased NADH:ferricyanide reductase activities, with kcat decreasing in the order WT > G179A > G179P > G179T > G179V, with the G179V variant retaining only 1.5% of the wild-type activity. The affinity for NADH also decreased in the order WT > G179A > G179P > G179T > G179V, with the Km(NADH) for G179V 180-fold greater than that of the wild type. Both Ks(H4NAD) and Ks(NAD+) values confirmed that the G179 mutants had both compromised NADH- and NAD+-binding affinities. Determination of the NADH/NADPH specificity constant for the various mutants indicated that G179 also participated in pyridine nucleotide selectivity, with the G179V variant preferring NADPH approximately 8000 times more than wild-type cb5r. These results demonstrated that, while G179 was not critical for either flavin incorporation or maintenance of the appropriate flavin environment in cb5r, G179 was required for both effective NADH/NADPH selectivity and to maintain the correct orientation and position of the conserved cysteine in the proline-rich "CGpppM" motif that is critical for optimum NADH binding and efficient hydride transfer. PMID- 16216071 TI - C-terminal tyrosine of ferredoxin-NADP+ reductase in hydride transfer processes with NAD(P)+/H. AB - Ferredoxin-NADP+ reductase (FNR) catalyzes the reduction of NADP+ to NADPH in an overall reversible reaction, showing some differences in the mechanisms between cyanobacterial and higher plant FNRs. During hydride transfer it is proposed that the FNR C-terminal Tyr is displaced by the nicotinamide. Thus, this C-terminal Tyr might be involved not only in modulating the flavin redox properties, as already shown, but also in nicotinamide binding and hydride transfer. FNR variants from the cyanobacterium Anabaena in which the C-terminal Tyr has been replaced by Trp, Phe, or Ser have been produced. All FNR variants show enhanced NADP+ and NAD+ binding, especially Tyr303Ser, which correlates with a noticeable improvement of NADH-dependent reactions. Nevertheless, the Tyr303Ser variant shows a decrease in the steady-state kcat value with NADPH. Fast kinetic analysis of the hydride transfer shows that the low efficiency observed for this mutant FNR under steady-state conditions is not due to a lack of catalytic ability but rather to the strong enzyme-coenzyme interaction. Three-dimensional structures for Tyr303Ser and Tyr303Trp variants and its complexes with NADP+ show significant differences between plant and cyanobacterial FNRs. Our results suggest that modulation of coenzyme affinity is highly influenced by the strength of the C-terminus-FAD interaction and that subtle changes between plant and cyanobacterial structures are able to modify the energy of that interaction. Additionally, it is shown that the C-terminal Tyr of FNR lowers the affinity for NADP+/H to levels compatible with steady-state turnover during the catalytic cycle, but it is not involved in the hydride transfer itself. PMID- 16216073 TI - Stability and flexibility in the structure of the hyperthermophile DNA-binding protein Sac7d. AB - Sac7d is a chromatin protein from the hyperthermophile Sulfolobus acidocaldarius that severely kinks duplex DNA with negligible change in protein structure. In previous work, the overall stability of Sac7d has been well-characterized with a global analysis of the linkage of folding, protonation, and anion binding. We extend that work here with NMR measurements of global stability as well as the distribution of stability and flexibility in the solution structure. Native state amide hydrogen exchange has been used to identify the most-protected core amide protons which exchange through global unfolding. The pH and temperature dependence of stability defined by native state exchange is in excellent agreement with the free energy surface determined by a linkage analysis of the dependence of folding on pH, salt, and temperature. These results confirm that the deltaC(P) obtained from a Kirchhoff analysis of DSC data (i.e., deltaH vs Tm) is incorrect, and an accurate description of the protein stability curve for Sac7d requires a measure of the thermodynamic contributions of protonation and anion binding. Amide hydrogen exchange, along with generalized order parameters determined by 15N relaxation data, demonstrates considerable variation in stability throughout the structure with some of the least stable regions occurring at the N- and C-termini. The most stable and inflexible region of the backbone occurs primarily in the DNA-binding beta-sheet which is responsible for bending DNA. PMID- 16216072 TI - Mechanisms of product feedback regulation and drug resistance in cytidine triphosphate synthetases from the structure of a CTP-inhibited complex. AB - Cytidine triphosphate synthetases (CTPSs) synthesize CTP and regulate its intracellular concentration through direct interactions with the four ribonucleotide triphosphates. In particular, CTP product is a feedback inhibitor that competes with UTP substrate. Selected CTPS mutations that impart resistance to pyrimidine antimetabolite inhibitors also relieve CTP inhibition and cause a dramatic increase in intracellular CTP concentration, indicating that the drugs act by binding to the CTP inhibitory site. Resistance mutations map to a pocket that, although adjacent, does not coincide with the expected UTP binding site in apo Escherichia coli CTPS [EcCTPS; Endrizzi, J. A., et al. (2004) Biochemistry 43, 6447-6463], suggesting allosteric rather than competitive inhibition. Here, bound CTP and ADP were visualized in catalytically active EcCTPS crystals soaked in either ATP and UTP substrates or ADP and CTP products. The CTP cytosine ring resides in the pocket predicted by the resistance mutations, while the triphosphate moiety overlaps the putative UTP triphosphate binding site, explaining how CTP competes with UTP while CTP resistance mutations are acquired without loss of catalytic efficiency. Extensive complementarity and interaction networks at the interfacial binding sites provide the high specificity for pyrimidine triphosphates and mediate nucleotide-dependent tetramer formation. Overall, these results depict a novel product inhibition strategy in which shared substrate and product moieties bind to a single subsite while specificity is conferred by separate subsites. This arrangement allows for independent adaptation of UTP and CTP binding affinities while efficiently utilizing the enzyme surface. PMID- 16216074 TI - Crystal structure analysis of phosphatidylcholine-GM2-activator product complexes: evidence for hydrolase activity. AB - GM2-activator protein (GM2AP) is a lysosomal lipid transfer protein with important biological roles in ganglioside catabolism, phospholipid metabolism, and T-cell activation. Previous studies of crystal structures of GM2AP complexed with the physiological ligand GM2 and platelet activating factor (PAF) have shown binding at two specific locations within the spacious apolar pocket and an ordering effect of endogenous resident lipids. To investigate the structural basis of phospholipid binding further, GM2AP was cocrystallized with phosphatidylcholine (PC), known to interact with GM2AP. Analysis of three crystal forms revealed binding of single chain lipids and fatty acids only and surprisingly not intact PC. The regions of best defined electron density are consistent with the presence of lyso-PC and oleic acid, which constitute deacylation products of PC. Their acyl tails are in stacking contact with shorter, less well-defined stretches of electron density that may represent resident fatty acids. The GM2AP associated hydrolytic activity that generates lyso-PC was further confirmed by mass spectrometry and enzymatic assays. In addition, we report the structures of (i) mutant Y137S, assessing the role of Tyr137 in lipid transfer via the hydrophobic cleft, and (ii) apo-mouse GM2AP, revealing a hydrophobic pocket with a constricted opening. Our structural results provide new insights into the biological functions of GM2AP. The combined effect of hydrolytic and lipid transfer properties has profound implications in cellular signaling. PMID- 16216075 TI - Calbindin D28k EF-hand ligand binding and oligomerization: four high-affinity sites--three modes of action. AB - Calbindin D28k, a highly conserved protein with Ca2+-sensing and Ca2+-buffering capabilities, is abundant in brain and sensory neurons. This protein contains six EF-hand subdomains, four of which bind Ca2+ with high affinity. Calbindin D28k can be reconstituted from six synthetic peptides corresponding to the six EF hands, indicating a single-domain structure with multiple interactions between the EF-hand subdomains. In this study, we have undertaken a detailed characterization of the Ca2+-binding and oligomerization properties of each individual EF-hand peptide using CD spectroscopy and analytical ultracentrifugation. Under the conditions tested, EF2 is monomeric and does not bind Ca2+, whereas EF6, which binds Ca2+ weakly, aggregates severely. We have therefore focused this study on the high-affinity binding sites, EF-hands 1, 3, 4, and 5. Our sedimentation equilibrium data show that, in the presence of Ca2+, EF-hands 1, 3, 4, and 5 all form dimers in solution in which the distribution between the monomer, dimer, and higher order oligomers differs. The processes of Ca2+ binding and oligomerization are linked to different degrees, and three main mechanisms emerge. For EF-hands 1 and 5, the dimer binds Ca2+ more strongly than the monomer and Ca2+ binding drives dimerization. For EF-hand 4, dimer formation requires only one of the monomers to be Ca2+-bound. In this case, the Ca2+ affinity is independent of dimerization. For EF-hand 3, dimerization occurs both in the absence and presence of Ca2+, while oligomerization increases in the presence of Ca2+. PMID- 16216076 TI - Crystal structure of the RIM2 C2A-domain at 1.4 A resolution. AB - RIMs are large proteins that contain two C2-domains and are localized at presynaptic active zones, where neurotransmitters are released. RIMs play key roles in synaptic vesicle priming and regulation of presynaptic plasticity. A mutation in the RIM1 C2A-domain has been implicated in autosomal dominant cone rod dystrophy (CORD7). The RIM C2A-domain does not contain the full complement of aspartate residues that commonly mediate Ca2+ binding at the top loops of C2 domains, and has been reported to interact with SNAP-25 and synaptotagmin 1, two proteins from the Ca2+-dependent membrane fusion machinery. Here we have used NMR spectroscopy and X-ray crystallography to analyze the structure and biochemical properties of the RIM2 C2A-domain, which is closely related to the RIM1 C2A domain. We find that the RIM2 C2A-domain does not bind Ca2+. Moreover, little binding of the RIM2 C2A-domain to SNAP-25 and to the C2-domains of synaptotagmin 1 was detected by NMR experiments, suggesting that as yet unidentified interactions of the RIM C2A-domain mediate its function. The crystal structure of the RIM2 C2A-domain using data to 1.4 A resolution reveals a beta-sandwich that resembles those observed for other C2-domains, but exhibits a unique dipolar distribution of electrostatic charges whereby one edge of the beta-sandwich is highly positive and the other edge is highly negative. The location of the mutation site implicated in CORD7 at the bottom of the domain and the pattern of sequence conservation suggest that, in contrast to most C2-domains, the RIM C2A domains may function through Ca2+-independent interactions involving their bottom face. PMID- 16216077 TI - Proteolytic action of GlpG, a rhomboid protease in the Escherichia coli cytoplasmic membrane. AB - We characterized Escherichia coli GlpG as a membrane-embedded protease and a possible player in the regulated intramembrane proteolysis in this organism. From the sequence features, it belongs to the widely conserved rhomboid family of membrane proteases. We verified the expected topology of GlpG, and it traverses the membrane six times. A model protein having an N-terminal and periplasmically localized beta-lactamase (Bla) domain, a LacY-derived transmembrane region, and a cytosolic maltose binding protein (MBP) mature domain was found to be GlpG dependently cleaved in vivo. This proteolytic reaction was reproduced in vitro using purified GlpG and purified model substrate protein, and the cleavage was shown to occur between Ser and Asp in a region of high local hydrophilicity, which might be located in a juxtamembrane rather than an intramembrane position. The conserved Ser and His residues of GlpG were essential for the proteolytic activities. Our results using several variant forms of the model protein suggest that GlpG recognizes features of the transmembrane regions of substrates. These results point to a detailed molecular mechanism and cellular analysis of this interesting class of membrane-embedded proteases. PMID- 16216078 TI - Metal binding characteristics and role of iron oxidation in the ferric uptake regulator from Escherichia coli. AB - The ferric uptake regulator is a metal-dependent transcription repressor that is activated by divalent transition metal cations. Fe(II) is believed to be the primary functional metal in vivo; however, the ability of other divalent cations to activate Fur brings into question the true physiological metal. Furthermore, the role of different oxidation states of iron in activating Fur has not been determined. Comparison of the affinity of different metals with intracellular metal concentrations would suggest which metals activate Fur in vivo; however, no accurate determinations of the affinity of Fur for metals have been reported. In this study, methods for reconstituting Fur with Fe(II), Fe(III), Co(II), and Zn(II) are described. Reconstituted protein was assayed for DNA affinity by gel shift assays. Fur is activated for DNA binding when reconstituted with Fe(III), as well as Fe(II), Zn(II), Co(II), and Mn(II), with little difference in DNA affinity for the different metallo forms of Fur. The affinity of Fur for the different metals was determined and ranges over several orders of magnitude in the following order: Zn(II) >> Co(II) > Fe(II) > Mn(II). Only Fe(II) binds with sufficient affinity to activate Fur significantly at physiological metal concentrations, when compared to previously determined total metal concentrations in Escherichia coli. PMID- 16216079 TI - Catalytic mechanism of fungal homoserine transacetylase. AB - Homoserine transacetylase is a required catalyst in the biochemical pathway that metabolizes Asp to Met in fungi. The enzyme from the yeast Schizosaccharomyces pombe activates the hydroxyl group of L-homoserine by acetylation from acetyl coenzyme A. This enzyme is unique to fungi and some bacteria and presents an important new target for drug discovery. Steady-state kinetic parameters provide evidence that this enzyme follows a ping-pong mechanism. Proton inventory was consistent with a single-proton transfer, and pH studies suggested the participation of at least one residue with a pKa value of 6.4-6.6, possibly a His or Asp/Glu in catalysis. Protein sequence alignments indicate that this enzyme belongs to the alpha/beta-hydrolase fold superfamily of enzymes, indicating the involvement of an active-site nucleophile and possibly a canonical catalytic triad. We constructed site-specific mutants and identified Ser163, Asp403, and His432 as the likely active-site residues of a catalytic triad based on steady state kinetics and genetic complementation of a yeast null mutant. Moreover, unlike the wild-type enzyme, inactive site mutants were not capable of producing an acetyl-enzyme intermediate. Homoserine transacetylase therefore catalyzes the acetylation of L-homoserine via a covalent acyl-enzyme intermediate through an active-site Ser. These results form the basis of future exploitation of this enzyme as an antimicrobial target. PMID- 16216080 TI - Transient formation of a neutral ubisemiquinone radical and subsequent intramolecular electron transfer to pyrroloquinoline quinone in the Escherichia coli membrane-integrated glucose dehydrogenase. AB - The membrane-bound quinoprotein glucose dehydrogenase (mGDH) in Escherichia coli contains pyrroloquinoline quinone (PQQ) and participates in the direct oxidation of D-glucose to D-gluconate by transferring electrons to ubiquinone (UQ). To elucidate the mechanism of ubiquinone reduction by mGDH, we applied a pulse radiolysis technique to mGDH with or without bound UQ8. With the UQ8-bound enzyme, a hydrated electron reacted with mGDH to form a transient species with an absorption maximum at 420 nm, characteristic of formation of a neutral ubisemiquinone radical. Subsequently, the decay of the absorbance at 420 nm was accompanied by an increase in the absorbance at 370 nm. Experiments with the PQQ free apoenzyme showed no such subsequent absorption changes, although ubisemiquinone was formed. These results indicate that a pathway for an intramolecular electron transfer from ubisemiquinone radical at the UQ8 binding site to PQQ exists in mGDH. The first-order rate constant of this process was calculated to be equal to 1.2 x 10(3) s(-1). These findings are consistent with our proposal that during the catalytic cycle of mGDH the bound UQ8 mediates electron transfer from the reduced PQQ to UQ8 pools. PMID- 16216081 TI - The carboxy-terminal tail of pyruvate dehydrogenase kinase 2 is required for the kinase activity. AB - Pyruvate dehydrogenase kinase 2 (PDK2) is a prototypical mitochondrial protein kinase that regulates the activity of the pyruvate dehydrogenase complex. Recent structural studies have established that PDK2 consists of a catalytic core built of the B and K domains and the relatively long amino and carboxyl tails of unknown function. Here, we show that the carboxy-terminal truncation variants of PDK2 display a greatly diminished capacity for phosphorylation of holo-PDC. This effect is due largely to the inability of the transacetylase component of PDC to promote the phosphorylation reaction catalyzed by the truncated PDK2 variants. Furthermore, the truncated forms of PDK2 bind poorly to the lipoyl-bearing domain(s) provided by the transacetylase component. Taken together, these data strongly suggest that the carboxyl tails of PDK isozymes contribute to the lipoyl bearing domain-binding site of the kinase molecule. We also show that the carboxyl tails derived from isozymes PDK1, PDK3, and PDK4 are capable of supporting the kinase activity of the kinase core derived from PDK2 as well as binding of the respective PDK2 chimeras to the lipoyl-bearing domain. Furthermore, the chimera carrying the carboxyl tail of PDK3 displays a stronger response to the addition of the transacetylase component along with a better binding to the lipoyl-bearing domain, suggesting that, at least in part, the differences in the amino acid sequences of the carboxyl tails account for the differences between PDK isozymes. PMID- 16216082 TI - Escherichia coli cyclopropane fatty acid synthase: is a bound bicarbonate ion the active-site base? AB - Cyclopropane synthases catalyze the cyclopropanation of unsaturated fatty acid using S-adenosyl-L-methionine as the methylene donor. The crystal structure of three cyclopropane synthases from Mycobacterium tuberculosis showed a bicarbonate ion bound in the active site that was proposed to act as a general base in the reaction mechanism [Huang, C., Smith, V., Glickman, M. S., Jacobs, W. R., and Sacchettini, J. C. (2002) J. Biol. Chem. 277, 11559-11569]. Because the in vitro activity of M. tuberculosis cyclopropane synthases has not yet been reported and because the ligands of the bicarbonate ion are all strictly conserved in cyclopropane synthases, we used the closely related Escherichia coli cyclopropane fatty acid synthase for this study. The putative ligands that share a hydrogen bond with the bicarbonate through their side chains were mutated. H266A, Y317F, E239A, and E239Q mutants were thus constructed and purified, and their catalytic efficiencies were 5.3, 0.7, 0.2, and <0.02%, respectively. C139 that is bound to the bicarbonate by its NH amide had already been mutated to serine in a previous work, and this mutant retains 31% of the activity of the wild-type enzyme. Kinetic analyses and binding studies using spectrofluorimetry showed that these mutations affected the catalytic constant rather than the binding of the substrates. While addition of free bicarbonate had almost no effect on the wild type enzyme activity, all mutants, with the exception of E239A and E239Q, were rescued by the addition of free bicarbonate. The catalytic efficiencies of the rescued mutants were 85, 16, and 14% for C139S, H266A, and Y317F, respectively. This effect was specific to bicarbonate. The kinetic parameters of the rescued mutants were determined, and it is shown that the rescuing effect is due to an increase in kcat. These data are interpreted by assuming that the E. coli cyclopropane fatty acid synthase specifically binds a bicarbonate ion that is involved in catalysis, as proposed for the M. tuberculosis enzymes, and that mutation of the bicarbonate ligands decreases the affinity for that ion. However, because the E239Q mutation could not be rescued, we propose that E239 forms a catalytic dyad with the bicarbonate to perform the proton abstraction necessary in the chemical pathway to the cyclopropane ring. PMID- 16216083 TI - ATP- and ADP-dependent modulation of RNA unwinding and strand annealing activities by the DEAD-box protein DED1. AB - DEAD-box RNA helicases, which are involved in virtually all aspects of RNA metabolism, are generally viewed as enzymes that unwind RNA duplexes or disrupt RNA-protein interactions in an ATP-dependent manner. Here, we show in vitro that the DEAD-box protein DED1 from Saccharomyces cerevisiae promotes not only RNA unwinding but also strand annealing, the latter in such a profound fashion that the physical limit for a bimolecular association rate constant is approached. We further demonstrate that DED1 establishes an ATP-dependent steady state between unwinding and annealing, which enables the enzyme to modulate the balance between the two opposing activities through ATP and ADP concentrations. The ratio between unwinding and annealing and the degree to which both activities are ATP- and ADP modulated are strongly influenced by structured as well as unstructured regions in the RNA substrate. Collectively, these findings expand the known functional repertoire of DEAD-box proteins and reveal the capacity of DED1 to remodel RNA in response to ADP and ATP concentrations by facilitating not only disruption but also formation of RNA duplexes. PMID- 16216084 TI - The 2F5 epitope is helical in the HIV-1 entry inhibitor T-20. AB - The HIV-1 envelope glycoprotein gp41 is responsible for viral fusion with the host cell. The fusion process, as well as the full structure of gp41, is not completely understood. One of the strongest inhibitors of HIV-1 fusion is a 36 residue peptide named T-20, gp41(638-673) (Fuzeon, also called Enfuvirtide or DP 178; residues are numbered according to the HXB2 gp160 variant) now used as an anti HIV-1 drug. This peptide also contains the immunogenic sequences that represent the full or partial recognition epitope for the broadly neutralizing human monoclonal antibodies 2F5 and 4E10, respectively. Due to its hydrophobicity, T-20 tends to aggregate at high concentrations in water, and therefore the structure of this molecule in aqueous solution has not been previously determined. We expressed a uniformly 13C/15N-labeled 42-residue peptide NN-T-20-NITN (gp41(636-677)) and used heteronuclear 2D and 3D NMR methods to determine its structure. Due to the additional gp41-native hydrophilic residues, NN-T-20-NITN dissolved in water, enabling for the first time determination of its secondary structure at near physiological conditions. Our results show that the NN-T-20-NITN peptide is composed of a mostly unstructured N terminal region and a helical region beginning at the center of T-20 and extending toward the C-terminus. The helical region is found under various conditions and has been observed also in a 13-residue peptide gp41(659-671). We suggest that this helical conformation is maintained in most of the different tertiary structures of the gp41 envelope protein that form during the process of viral fusion. Accordingly, an important element of the immunogenicity of gp41 and the inhibitory properties of Fuzeon may be the propensity of specific sequences in these polypeptides to assume helical structures. PMID- 16216085 TI - Disruption of the interaction of alpha-synuclein with microtubules enhances cell surface recruitment of the dopamine transporter. AB - Mutations in alpha-synuclein have been implicated in the genesis of Parkinson's disease. A probable normative function of alpha-synuclein is the maintenance of dopamine homeostasis, partly through a negative modulation of dopamine transporter (DAT) activity, by reducing its level at the cell surface. To study the possible involvement of the microtubular network in the alpha-synuclein dependent trafficking of DAT, we treated cotransfected cells and primary mesencephalic neurons with either colchicine, vinblastine, or nocodazole, each of which disrupts microtubules or affects microtubule dynamics. Treatment of both types of cells with vinblastine, colchicine, or nocodazole reversed alpha synuclein-mediated inhibition of DAT activity, resulting in an increased rate of dopamine uptake and and increased level of extracellular dopamine-induced oxidative stress, with accelerated cell death. Treatment with these agents also reversed the alpha-synuclein-induced decrease in levels of cell surface associated DAT. This effect of colchicine, vinblastine, or nocodazole was not linked to a disruption of formation of the alpha-synuclein-DAT complex but paradoxically caused an increased level of interaction between these proteins. Both alpha-synuclein and DAT co-immunoprecipitated with both alpha- and beta tubulins, in both transfected cells and rat primary mesencephalic dopaminergic neurons, suggesting heteromeric complex formation between these various proteins. Treatment with the microtubule depolymerizing agents disrupted the heteromeric protein complex between either alpha-synuclein or the DAT, and alpha- or beta tubulins. These results indicate a previously unappreciated role of microtubules in the modulation of DAT trafficking, and provide insight into a novel mechanism by which alpha-synuclein regulates DAT activity, by tethering the transporter to the microtubular network. PMID- 16216086 TI - The geographic spread of the CCR5 Delta32 HIV-resistance allele. AB - The Delta32 mutation at the CCR5 locus is a well-studied example of natural selection acting in humans. The mutation is found principally in Europe and western Asia, with higher frequencies generally in the north. Homozygous carriers of the Delta32 mutation are resistant to HIV-1 infection because the mutation prevents functional expression of the CCR5 chemokine receptor normally used by HIV-1 to enter CD4+ T cells. HIV has emerged only recently, but population genetic data strongly suggest Delta32 has been under intense selection for much of its evolutionary history. To understand how selection and dispersal have interacted during the history of the Delta32 allele, we implemented a spatially explicit model of the spread of Delta32. The model includes the effects of sampling, which we show can give rise to local peaks in observed allele frequencies. In addition, we show that with modest gradients in selection intensity, the origin of the Delta32 allele may be relatively far from the current areas of highest allele frequency. The geographic distribution of the Delta32 allele is consistent with previous reports of a strong selective advantage (>10%) for Delta32 carriers and of dispersal over relatively long distances (>100 km/generation). When selection is assumed to be uniform across Europe and western Asia, we find support for a northern European origin and long range dispersal consistent with the Viking-mediated dispersal of Delta32 proposed by G. Lucotte and G. Mercier. However, when we allow for gradients in selection intensity, we estimate the origin to be outside of northern Europe and selection intensities to be strongest in the northwest. Our results describe the evolutionary history of the Delta32 allele and establish a general methodology for studying the geographic distribution of selected alleles. PMID- 16216087 TI - Enhanced odor discrimination and impaired olfactory memory by spatially controlled switch of AMPA receptors. AB - Genetic perturbations of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptors (AMPARs) are widely used to dissect molecular mechanisms of sensory coding, learning, and memory. In this study, we investigated the role of Ca2+ permeable AMPARs in olfactory behavior. AMPAR modification was obtained by depletion of the GluR-B subunit or expression of unedited GluR-B(Q), both leading to increased Ca2+ permeability of AMPARs. Mice with this functional AMPAR switch, specifically in forebrain, showed enhanced olfactory discrimination and more rapid learning in a go/no-go operant conditioning task. Olfactory memory, however, was dramatically impaired. GluR-B depletion in forebrain was ectopically variable ("mosaic") among individuals and strongly correlated with decreased olfactory memory in hippocampus and cortex. Accordingly, memory was rescued by transgenic GluR-B expression restricted to piriform cortex and hippocampus, while enhanced odor discrimination was independent of both GluR-B variability and transgenic GluR-B expression. Thus, correlated differences in behavior and levels of GluR-B expression allowed a mechanistic and spatial dissection of olfactory learning, discrimination, and memory capabilities. PMID- 16216089 TI - Endovascular therapy of inflammatory aortic aneurysms: a meta-analysis. AB - PURPOSE: To report a meta-analysis of results from endovascular aneurysm repair (EVAR) of inflammatory aortic aneurysms (IAA). METHODS: A comprehensive literature review was performed to identify all studies reporting the results of EVAR in patients with IAA. To be included in the analysis, an article had to provide a minimum follow-up of 6 months, information about primary technical success, details of immediate and long-term complications, and evaluation of at least one of the basic outcome criteria: changes in aneurysm sac diameter, periaortic fibrosis (PAF), and/ or renal impairment. All studies were reviewed by 2 independent observers for the inclusion criteria. Data were retrieved on the technical and clinical success, outcome criteria, mortality in follow-up, and reinterventions from 14 articles selected from among 701 initially identified. RESULTS: The 14 articles encompassed 46 patients (45 men [97.8%]; mean age 65 years, range 59-75) with a mean follow-up of 18 months after endovascular repair of IAAs located in the abdominal aorta. The primary technical success rate was 95.6% (44/46) and the 30-day clinical success rate was 93.4% (43/46). The median aneurysm sac diameter regression was 11 mm. Of 43 patients with PAF prior to the intervention, 22 (51.2%) patients showed complete regression, 18 (41.8%) remained unchanged, and 3 (7.0%) showed progression after EVAR. Renal impairment disappeared in 11 (45.8%) of 24 patients. Reinterventions were reported in 8 patients. The procedure-related and follow-up mortality rates were 0% and 13.0%, respectively. CONCLUSIONS: EVAR of IAA is feasible, excludes the aneurysm effectively, and reduces PAF and renal impairment in most patients with very low periprocedural and midterm mortality and an acceptable reintervention rate. PMID- 16216090 TI - Getting the balance right. PMID- 16216088 TI - Rapid behavioral and genomic responses to social opportunity. AB - From primates to bees, social status regulates reproduction. In the cichlid fish Astatotilapia (Haplochromis) burtoni, subordinate males have reduced fertility and must become dominant to reproduce. This increase in sexual capacity is orchestrated by neurons in the preoptic area, which enlarge in response to dominance and increase expression of gonadotropin-releasing hormone 1 (GnRH1), a peptide critical for reproduction. Using a novel behavioral paradigm, we show for the first time that subordinate males can become dominant within minutes of an opportunity to do so, displaying dramatic changes in body coloration and behavior. We also found that social opportunity induced expression of the immediate-early gene egr-1 in the anterior preoptic area, peaking in regions with high densities of GnRH1 neurons, and not in brain regions that express the related peptides GnRH2 and GnRH3. This genomic response did not occur in stable subordinate or stable dominant males even though stable dominants, like ascending males, displayed dominance behaviors. Moreover, egr-1 in the optic tectum and the cerebellum was similarly induced in all experimental groups, showing that egr-1 induction in the anterior preoptic area of ascending males was specific to this brain region. Because egr-1 codes for a transcription factor important in neural plasticity, induction of egr-1 in the anterior preoptic area by social opportunity could be an early trigger in the molecular cascade that culminates in enhanced fertility and other long-term physiological changes associated with dominance. PMID- 16216091 TI - A prospective study of the use of nitrous oxide inhalation sedation for dental treatment in anxious children. AB - AIM: To determine baseline data in relation to procedures undertaken during nitrous oxide inhalation sedation sessions within the Hospital Dental Service. STUDY DESIGN: A prospective study. METHODS: Data was collected over a twelve month period for patients attending the Department of Paediatric Dentistry, Dundee Dental Hospital, Dundee, Scotland, for dental treatment using nitrous oxide inhalation sedation. Overall behaviour and the outcome of treatment were assessed by the dentist providing sedation using the Frankl and Houpt Behaviour Rating Scales respectively. RESULTS: Data was available for 312 patients (F:169; M:143) with a median age of 11.0 (inter-quartile range 8.8, 12.7) years. Overall, 93% of patients successfully completed treatment using sedation. The majority of treatments comprised dental extractions; 19.8% and 41.3% were primary and permanent tooth extractions respectively. Regarding permanent teeth, nearly 50% were first permanent molar extractions compared with just over 30% for first premolars. Nearly three-quarter of cases were treated using a mixture of 30% nitrous oxide and 70% oxygen, with a median sedation time of 35.7 minutes. Only 7% of cases failed to commence or complete treatment and overall, there was a greater failure rate amongst visiting Community Dental Officers, compared with Hospital-based clinicians. CONCLUSION: Extraction of first permanent molar teeth can be successfully achieved using inhalation sedation. There is both a need for further postgraduate training in sedation techniques as well as the necessity to train further Specialists in Paediatric Dentistry to undertake care within the Community Dental Service. PMID- 16216092 TI - Enamel defects in children with coeliac disease. AB - AIM: This was to investigate the prevalence of enamel developmental defects in a group of children with a history of coeliac disease. METHODS: A study group of children attending the Dept. Paediatrics (Leeds General Infirmary), born between 1985 and 1986 and subsequently diagnosed and treated for coeliac disease (CD) were recruited. A group of age/sex-matched children attending the Paediatric Dentistry department were used as a control group (Cont). Examinations were carried out for enamel defects and opacities (DDE index), dmf, dmfs, DMF and DMFS (BASCD method), and a full medical and dental history were obtained. RESULTS: Significantly more children in the CD group had a greater number of enamel defects than controls for both primary (p=<0.02) and permanent (p=<0.001) dentitions. Opacities in both primary and permanent teeth were statistically significantly greater in the CD group than controls (p=<0.04 and p=<0.001 respectively). Dental caries in both primary and permanent dentitions was less in the CD group compared with the control group of children. CONCLUSION: Coeliac disease was associated with an increased prevalence of developmental enamel defects. PMID- 16216093 TI - Evaluation of three pulpotomy medicaments in primary teeth. AB - AIM: To examine the success of the one-appointment pulpotomy technique with three different medicaments on primary molar teeth. METHODS: The study was conducted on 104 primary molars in 104 children with an indication for pulpotomy treatment on at least one primary molar. Primary teeth were treated with either formocresol (FC) (34 teeth), calcium hydroxide (CA) (33 teeth) or ferric sulphate (FS) (37 teeth) using standardised criteria for the pulpotomy procedures. Teeth were clinically and radiographically evaluated during the examination period of 18 months. RESULTS: The clinical success rate at 18 months for the FC and FS groups was 90.9% and 89.2%, respectively. The CA showed an overall clinical success rate of 82.3%, with no statistical difference compared with either the FC or FS groups. The overall radiographic success for each technique was: FC 84.8%, CA 76.5%, and FS 81.1%. The presence of a dentine bridge above the pulp amputation site was observed radiographically for CA (47%), and FS (40.5%) pulpotomies without any statistical difference. Radiographic examination did not reveal the presence of a dentine bridge for any of the teeth treated with FC pulpotomy. CONCLUSION: Favourable clinical and radiographic success rates of ferric sulphate pulpotomy, comparable to formocresol were obtained. Therefore, ferric sulphate can be recommended as a pulpotomy medicament. PMID- 16216094 TI - Asthma inhalers and subsurface enamel demineralisation: an in situ pilot study. AB - AIM: The purpose of this pilot study was to identify the subsurface enamel demineralising potential of two possible acidogenic lactose-based powders and their corresponding generic pump inhalers. METHODS: Ten healthy non-asthmatic adults participated in a 5- leg randomised crossover study including a 10% sucrose control. A twice-daily 400 microg dose of inhaler was applied in vitro to a demineralised enamel slab on the buccal flange of a mandibular removable appliance before in situ placement for 14 days each. Lesion parameters were determined using transverse microradiography and digitised image analysis. RESULTS: Minimal demineralisation occurred with sucrose, both pump and one powder inhaler. The remaining powder was associated with remineralisation (p = 0.29). Overall, mean lesion depth increased (p = 0.12). CONCLUSION: Asthma inhalers failed to demonstrate a significant acidogenic/cariogenic effect. PMID- 16216095 TI - Parental presence versus absence in the dental operatory: a technique to manage the uncooperative child dental patient. AB - AIM: Societal changes have led to an increased parental presence in paediatric dental surgeries. The aim of this study was to assess the deliberate use of a parental presence/absence (PPA) intervention as a behavioural technique to manage very uncooperative children. STUDY DESIGN: This is a retrospective study of the PPA technique in a Greek paediatric dental practice. METHODS: Practice patient records were examined over a six year period, during which time Frankl behaviour ratings were made for each patient. At the initial dental visit, 85 otherwise normal healthy children were found to be very uncooperative (Frankl score of definitely negative); 46 female and 39 male, mean age 4.6+/-1.5 yrs (range 2.7 8.8). Their parents stepped out of the operatory and returned after the child became cooperative. These children were compared with 85 randomly selected cooperative children, matched for age and gender. STATISTICS: A one sample binomial test was used to assess the technique's success, while chi(2) and t tests were used for between group comparisons of proportions and means, respectively. RESULTS: Seventy-four out of the 85 initially uncooperative children displayed Frankl positive behaviour as the initial appointment progressed, while 6 children remained uncooperative during the first appointment but were cooperative at following visits. The total technique success was 80/85 (94%, p<0.001). The two groups differed significantly with regards to control in caries scores (frequency 77.6% with 12.2 +/- 11.8 dmfs as opposed to 54.1% with 6.7 +/- 5.7 dmfs), and in parent's awareness of patient dental needs or symptoms (76 vs 52). There was no significant difference between groups with regards to completing the treatment planned (63/80 vs 75/85). There was no significant difference in the proportion of returning children who displayed uncooperative behaviour in subsequent visits. CONCLUSIONS: The PPA technique can be successful in gaining the cooperation of children displaying very negative behaviour, thus minimising the need for other more aversive techniques. PMID- 16216096 TI - Efficacy of chlorhexidine varnish applications in the prevention of early childhood caries. AB - AIM: A prospective clinical study was conducted to evaluate the influence of 3 monthly applications of the chlorhexidine- containing varnish Cervitec on the colonisation of a child's oral cavity by mutans streptococci (MS) and on caries prevalence. METHODS: 200 children aged 11.7 +/- 0.7 months were examined. Children's caries risk was assessed on the basis of their MS scores in saliva. Out of the 48 children in whom a high caries risk (> or =10(5) CFU/ml) was expected, 23 were treated with Cervitec at 3-month intervals (CHX group). The remaining 25 children of the high-risk group received no treatment and served as controls (group C). All parents received detailed information on the prevention of early childhood caries. 172 children had completed the study after one year. RESULTS: During the course of the study the percentage of children with visible plaque on their maxillary incisors increased from 17.8% to 40.1% and the percentage of children given sweetened drinks in nursing bottles for the night rose from 16.3% to 18.0%. At the closure of the study 26.2% of the two-year-olds had salivary scores of MS > or =10(5) CFU/ml of saliva. The mean d(1-4)mft value increased from 0.05 -/+ 0.4 to 0.8 -/+ 2.9 and the mean d(1-4)mfs value rose from 0.08 -/+ 0.8 to 1.8 -/+ 5.9. No significant differences were demonstrable between the two-year-olds in groups CHX and C for colonisation of the oral cavity by MS or for d(1-4)mft values. In contrast the d(1-4)mfs values were significant lower in the CHX group as in the group C. CONCLUSION: Poor feeding habits and deficits in oral hygiene cannot be compensated by the application of Cervitec. PMID- 16216097 TI - Primary tooth fluorosis in 5-year-old schoolchildren in Ireland. AB - AIM: This was to determine the prevalence of primary tooth fluorosis in the dentitions of 5-year-old schoolchildren. A subsidiary aim was to investigate whether an association existed between the presence of primary tooth fluorosis, fluoridation status, infant feeding practices or the oral hygiene practices of the child. STUDY DESIGN: A cross-sectional and stratified by fluoridation status study. METHODS: Fluorosis was recorded using a modification of the Tooth Surface Index of Fluorosis (TSIF). Demographic data, information on infant feeding practices and oral hygiene practices were collected via a parental questionnaire. STATISTICS: Stepwise logistic regression analysis. RESULTS: Fluorosis prevalence in the fluoridated group (n=208) was 32%; 29.3% (n=61) had a modified TSIF score of 1; 2.4% (n=5) had a modified TSIF score of 2; and 1% (n=1) had a modified TSIF score of 5. In the non-fluoridated group (n=86) one child had a modified TSIF score of 1. Primary tooth prevalence of fluorosis in the entire sample (n=294) was 23%. Factors that were associated with primary tooth fluorosis were: fluoridation status (p= 0.0003, 95% CI = 5-281) and the age at which toothbrushing with toothpaste commenced (p = 0.016, 95% C.I. 1.1 - 3.8). No association with infant feeding practices was identified. CONCLUSION: The overall prevalence of primary tooth fluorosis was 23%. Lifetime residence in a fluoridated area and commencement of toothbrushing with toothpaste between 12 and 18 months of age were associated with primary tooth fluorosis. No association with infant feeding practices was identified. PMID- 16216098 TI - Infection with Fasciola hepatica. AB - Fascioliasis, caused by the liver fluke Fasciola hepatica, is an infection that occurs worldwide, although humans are accidental hosts. F. hepatica infection comprises two stages, hepatic and biliary, with different signs and symptoms. Stool examination and ELISA can be used for the initial diagnosis. Radiographic techniques, such as computerised tomography and ultrasonography, as well as magnetic resonance imaging, are used widely for confirmation and follow-up of the disease. Invasive techniques, such as percutaneous cholangiography, endoscopic retrograde cholangiography and liver biopsy, may aid in the diagnosis but are not essential. Triclabendazole is recommended as the first-line agent for the treatment of F. hepatica infection, with bithionol as an alternative. PMID- 16216099 TI - Irrigation with antibiotic-containing solutions for the prevention and treatment of infections. AB - Administration routes for antimicrobial agents used in clinical practice include the topical, inhaled, enteral and parenteral routes. An antibiotic administration route used frequently worldwide, although not well-studied, involves the irrigation of wounds with antibiotic-containing solutions for the prevention and treatment of infections. This review considers the data available from various experimental and clinical studies in order to provide an update on the use of antibiotic-containing solutions in modern clinical practice. Although irrigation with antibiotic-containing solutions has been suggested to be beneficial in the prevention or treatment of infections in several settings and patient populations, no firm, evidence-based recommendations can be made regarding its use until additional data from well-designed, randomised clinical trials become available. Current exceptions include empyema following lobectomy, or pneumonectomy and pyocystis (vesical empyema), since irrigation with solutions containing antimicrobial agents seems to be a crucial component of the management of these conditions. PMID- 16216100 TI - Clinical impact and pathogenicity of Acinetobacter. AB - Members of the genus Acinetobacter have been implicated in a wide spectrum of infectious diseases. Although this organism is associated primarily with nosocomial infections, it has also been involved in cases of community-acquired infection. Before the 1970s, Acinetobacter infections were mostly post-surgical urinary tract infections in patients hospitalised in surgical units. The significant improvement in resuscitation techniques during the last 30 years has changed the types of infection caused by Acinetobacter. Since the 1980s, Acinetobacter has spread rapidly among patients in intensive care units. Today, Acinetobacter accounts for c. 9% of nosocomial infections, with most Acinetobacter infections involving the respiratory tract. Transmission via the hands of hospital staff has become the most important contributory factor in patient colonisation. Acinetobacter baumannii is the species that is involved most frequently in infections of humans, but a natural reservoir for A. baumannii outside the hospital environment has not yet been identified. Community-acquired infection and infections acquired following war or natural disasters (e.g., earthquakes) have been described. Acinetobacter causes mild-to-severe illness, but can be fatal. The severity of Acinetobacter infection depends upon the site of infection and the patient's susceptibility to infection as a result of underlying disease. The circumstances that allow Acinetobacter to assume a pathogenic role are not really well-understood. As this organism is a low-grade pathogen, the pathogenesis of Acinetobacter infections probably involves numerous factors, including virulence determinants, which have yet to be investigated. PMID- 16216101 TI - Risk-factors for the acquisition of imipenem-resistant Acinetobacter baumannii in Spain: a nationwide study. AB - Potential risk-factors for the acquisition of imipenem-resistant Acinetobacter baumannii were investigated in a cohort study in 25 Spanish hospitals. The clonal relationship among isolates was determined by pulsed-field gel electrophoresis (PFGE). In total, A. baumannii was isolated from 203 patients, with imipenem resistant (MIC(90) 128 mg/L) isolates being obtained from 88 patients (43%), and imipenem-susceptible isolates from 115 patients (57%). A wide clonal distribution was observed among the imipenem-resistant isolates, but spread of the same clone among centres was not demonstrated. The results indicated that imipenem-resistant A. baumannii is a widely distributed nosocomial pathogen in Spain and reaches an alarming frequency in some centres. Independent risk-factors for the acquisition of imipenem-resistant A. baumannii were a hospital size of >500 beds (multivariate OR, 6.5; 95% CI, 1.8--23), previous antimicrobial treatment (multivariate OR, 4.3; 95% CI, 1.6--11), a urinary catheter (multivariate OR, 2.7; 95% CI, 1.1--6.7) and surgery (multivariate OR, 2; 95% CI, 1.07--3.8). PMID- 16216102 TI - Stenotrophomonas maltophilia: antimicrobial resistance and molecular typing of an emerging pathogen in a Turkish university hospital. AB - Despite its limited pathogenicity, Stenotrophomonas maltophilia is an emerging nosocomial pathogen. This study investigated the isolation frequency, antimicrobial resistance and genotypic relationships of 205 S. maltophilia isolates from 188 patients in a university hospital between 1998 and 2003. Susceptibility profiles for 11 antimicrobial agents were determined by the NCCLS agar dilution method for non-fermentative bacteria, while enterobacterial repetitive intergenic consensus sequence (ERIC)-PCR and pulsed-field gel electrophoresis (PFGE) were used for genotyping of the isolates. Of the 205 isolates, 56.1% were isolated in the last 2 years of the study. The risk of S. maltophilia isolation was higher in intensive care units, S. maltophilia was isolated mostly (86.8%) after hospitalisation for >or= 48 h, and 90.4% of the patients had underlying diseases. Resistance levels were>60% for all antimicrobial agents tested except co-trimoxazole. High genetic diversity was found among the S. maltophilia isolates, and cross-infection with S. maltophilia was not common. Although ERIC-PCR revealed fewer genotypes than PFGE, it proved to be a rapid and easy method for S. maltophilia genotyping, and was more economical than PFGE. PMID- 16216103 TI - Contribution of clonal dissemination and selection of mutants during therapy to Pseudomonas aeruginosa antimicrobial resistance in an intensive care unit setting. AB - Rates of antibiotic resistance in Pseudomonas aeruginosa isolates from intensive care unit (ICU) patients are expected to be dependent on the selection of resistance mutations during therapy, the availability of exogenous resistance determinants and their dissemination potential, and the efficiency of transmission of the resistant strains. The relative contributions of these three factors were studied in an ICU with no apparent outbreak in 216 sequential P. aeruginosa isolates recovered from 102 patients between September 2002 and November 2003. Analysis of pulsed-field gel electrophoresis patterns revealed the presence of 82 different clones. Thus, the dissemination of particular resistant clones had a minimal effect on the relatively high overall resistance frequencies found for imipenem (32%), cefepime (25%), ceftazidime (24%), meropenem (22%), ciprofloxacin (18%) and tobramycin (2%). Rates of primary resistance were relatively low, and resistance development during treatment (secondary resistance) was the main factor contributing to the overall high resistance rates. In ICU settings with a low prevalence of epidemic resistant strains, the main strategy for resistance control should focus on the design of targeted regimens to avoid the development of resistance. PMID- 16216104 TI - Identification of a novel cephalosporinase (DHA-3) in Klebsiella pneumoniae isolated in Taiwan. AB - A strain of Klebsiella pneumoniae resistant to cefoxitin and oxyimino cephalosporins, but susceptible to cefepime, was isolated from an adult patient hospitalised in Taichung, Taiwan. Isoelectric focusing revealed three beta lactamases with isoelectric points of 5.4, 8.2 and 7.9, respectively. Following PCR with plasmid DNA templates and gene sequencing, these enzymes were shown to correspond to TEM-1, SHV-5 and a novel DHA-1-like enzyme (designated DHA-3). The bla genes for TEM-1 and SHV-5 were transferable, but the bla(DHA-3) gene was non self-transferable in conjugation experiments. All three bla genes were successfully introduced by electrotransformation into an Escherichia coli recipient (DH5alpha), resulting in a similar resistance profile to that observed in the original donor strain. Other K. pneumoniae strains producing DHA-1-like enzymes have been identified previously in Taiwan, and this report suggests that DHA-type beta-lactamases are continuing to emerge in this country. PMID- 16216105 TI - Class 1 integrons in ciprofloxacin-resistant Escherichia coli strains from two Dutch hospitals. AB - A significant increase in the isolation frequency of ciprofloxacin-resistant Escherichia coli was observed in the haematology departments of two university hospitals in The Netherlands. Amplified fragment length polymorphism analysis revealed that this increase was not caused by the emergence of unique ciprofloxacin-resistant clones. Determination of the presence of class 1 integrons indicated that 81% of the ciprofloxacin-resistant isolates contained an intI1 gene, compared with 11% of the ciprofloxacin-susceptible isolates (p<0.0001). The quinolone resistance gene qnrA was not present in any of the integrons characterised and could not be detected using dot-blot hybridisation of total DNA. In addition, conjugation experiments showed that ciprofloxacin resistance was not co-transferred with class 1 integrons. Ciprofloxacin-resistant isolates harboured mutations in the gyrA gene, which are known to encode ciprofloxacin resistance. In conclusion, an association was observed between ciprofloxacin resistance and the presence of class 1 integrons, which could not be explained by the currently known genetic determinants of quinolone resistance. PMID- 16216107 TI - Secular trends in nosocomial candidaemia in non-neutropenic patients in an Italian tertiary hospital. AB - A retrospective study was performed in an Italian tertiary hospital to evaluate trends in candidaemia between 1992 and 2001, and to compare the characteristics of episodes of fungaemia between 1992--1997 and 1998--2001. In total, 370 episodes of candidaemia were identified, with an average incidence of 0.99 episodes/10 000 patient-days/year (range 0.49--1.29 episodes). On an annual trend basis, the overall incidence was essentially stable in surgical and medical wards, but decreased in intensive care units (ICUs) (p 0.0065). The average use of fluconazole was 37.9 g/10 000 patient-days/year (range 21.4--56.1 g), and did not change significantly during the 10-year period. Nearly two-thirds of patients were in ICUs at the onset of candidaemia, but none was neutropenic in either study period. Candida albicans remained the predominant species isolated (53.8% vs. 48.1%), followed by Candida parapsilosis, Candida glabrata and Candida tropicalis, the distribution of which did not change significantly. The 30-day crude mortality rate was essentially similar (44% vs. 35%) in both study periods. Thus the incidence of nosocomial candidaemia, although high in this institution, decreased among critically-ill patients during the 10-year period. This finding seemed to be related to an improvement in infection control practices, particularly regarding the prevention of intravascular catheter-related infections in ICUs. Although the overall use of fluconazole was considerable, no increase in azole-resistant non-albicans Candida spp. was detected. PMID- 16216106 TI - Performance of the Now Malaria rapid diagnostic test with returned travellers: a 2-year retrospective study in a French teaching hospital. AB - Malaria caused by Plasmodium falciparum remains the major life-threatening parasitic infection in the world. The number of cases in non-endemic countries continues to increase, and it is important that misdiagnosis of malaria should not occur, especially in non-immune travellers, because of the high risk of a fatal outcome. In a retrospective study of 399 sera, the Now Malaria rapid test was compared with the quantitative buffy coat (QBC) test and microbiological examination of thin blood films. Compared with the QBC test and thin blood films, the Now Malaria test had sensitivity and specificity values of 96.4% and 97%, respectively, for the detection of pure P. falciparum infection. A negative predictive value of 99.4% allows this test to be included in diagnostic strategies for patients presenting with clinical suspicion of malaria. Two false negative results were associated with low levels of parasitaemia in the specimens. Thus, use of the Now Malaria test alone to detect P. falciparum infection in non-endemic countries could lead to misdiagnosis of malaria. This rapid diagnostic test should therefore be performed in association with another prompt traditional method such as examination of thin blood films. PMID- 16216108 TI - Aetiology of sexually transmitted disease (STD) and comparison of STD syndromes and aetiological diagnosis in Taipei, Taiwan. AB - Between July 2002 and July 2004, all male patients (n=307) presenting to a sexually transmitted disease (STD) clinic who reported unprotected sex were enrolled in the study. Of these, 63 (20.5%), 19 (6.2%) and one (0.3%) were infected with one, two and three pathogens, respectively. The prevalences of chlamydial infection, gonorrhoea, syphilis, human immunodeficiency virus infection and trichomoniasis were 14.3%, 10.1%, 6.8%, 2.2% and 0%, respectively. The sensitivities of urethral discharge syndrome for detection of chlamydial, gonococcal, and combined forms of infection were 31.8%, 58.5% and 70.0%, respectively, with specificities of 93.5%, 97.1% and 93.9%, respectively. Positive predictive values (PPVs) were 56.0%, 68.0% and 28.0% for chlamydial and gonococcal infections and combinations of the two forms, respectively. In contrast, the sensitivity of genital ulceration syndrome for syphilis detection was only 38.0%, although the specificity was 82.5%, and the PPV was 32.0%. To reduce the number of false-positive results for STD patients who practise unprotected sex, incorporation of risk assessment and rapid diagnostic tests are recommended. PMID- 16216109 TI - Nosocomial bloodstream infections caused by Streptococcus pneumoniae. AB - A retrospective study of Streptococcus pneumoniae bacteraemia among adult patients in two large teaching hospitals in Spain identified 108 (10.6%) of 1,020 episodes as nosocomial pneumococcal bloodstream infections (NPBIs). Seventy-seven clinical records with sufficient data were available for analysis. The interval between admission and a positive blood culture was 3--135 days (median 17 days; interquartile range 8--27). The main underlying and predisposing conditions for NPBI were malignancy (31%), chronic obstructive pulmonary disease (28.6%), heart failure (16.9%), chronic renal failure (15.6%), liver cirrhosis (13%) and infection with human immunodeficiency virus (13%). Overall, 31.2% of patients developed severe sepsis, 11.7% septic shock, and 3.9% multi-organ failure. The main portals of entry were pneumonia (70.1%), meningitis (5.2%) and primary peritonitis (5.2%). Of the responsible serogroups, 78% were included in the 23 valent polysaccharide vaccine. Thirty-five (45.5%) patients died, with death considered to be related to the NPBI in 21 (27.3%) cases. Following multivariate analysis, factors that independently predicted death after adjusting for age were: ultimately fatal underlying disease (OR, 8.9; 95% CI, 0.8--94.3; p<0.001); rapidly fatal underlying disease (OR, 15.0; 95% CI, 2.8--81.3; p<0.001); heart failure (OR, 8.11; 95% CI, 1.1--60.8; p<0.03); inadequate empirical therapy (OR, 10.6; 95% CI, 1.2--97; p<0.003); a severe sepsis score (OR, 9.5; 95% CI, 1.9- 47.0; p<0.001); and septic shock or multi-organ failure (OR, 63.7; 95% CI, 4.9- 820.7; p<0.001). Adequate empirical therapy was an independent protective factor (OR, 0.05; 95% CI, 0.04--0.58; p<0.005), but the use of more than one antimicrobial agent was not. PMID- 16216110 TI - Decreasing rates of resistance to penicillin, but not erythromycin, in Streptococcus pneumoniae after introduction of a policy to restrict antibiotic usage in Taiwan. AB - A 16% decline in rates of penicillin resistance in Streptococcus pneumoniae isolates in Taiwan between 1998--1999 (25%) and 2001 (9%) was associated with a 46% decrease in total penicillin and other cephalosporin usage in 2001 compared with 1999. However, erythromycin resistance in S. pneumoniae remained high (94%), despite a 45% decrease in macrolide consumption between 1999 and 2001. PMID- 16216111 TI - Distribution of mef(A)-containing genetic elements in erythromycin-resistant isolates of Streptococcus pyogenes from Italy. AB - In total, 124 Streptococcus pyogenes isolates were obtained from throat cultures of different symptomatic patients. All isolates showed M-phenotype macrolide resistance and contained the macrolide efflux gene mef(A). The isolates were screened for the presence and insertion site of mef(A)-containing genetic elements. In 25.8% of the isolates, mef(A) was found to be carried by elements belonging to the Tn1207.3/Phi10394.4 family inserted in the comEC gene, while 74.2% contained chimeric elements with a different genetic structure and chromosomal location, probably associated with the recently described 60-kb tet(O)-mef(A) element. PMID- 16216112 TI - Rapid identification of Burkholderia pseudomallei in blood culture supernatants by a coagglutination assay. AB - In total, 309 blood culture supernatants were tested for the presence of Burkholderia pseudomallei antigen using an in-house coagglutination test prepared by sensitising Cowan I staphylococcal cells with B. pseudomallei polyclonal antiserum. The coagglutination test gave a sensitivity, specificity, positive predictive value and negative predictive value of 100% in comparison with blood culture. A subset of 102 supernatants was also tested for B. pseudomallei antigen using a monoclonal antibody-based latex agglutination test. The sensitivity, specificity, positive predictive value and negative predictive values of this test were 100%, 90%, 75% and 100%, respectively. PMID- 16216113 TI - Clinical and socio-economic impact of influenza and respiratory syncytial virus infection on healthy children and their households. AB - This prospective study compared the clinical and socio-economic impact of laboratory-confirmed influenza and respiratory syncytial virus (RSV) infection on healthy children and their families. Among 1,520 otherwise healthy children aged< 15 years attending the Emergency Department for acute conditions other than trauma, influenza viruses and RSV were found in 234 (15.4%) and 116 (7.6%; p<0.0001) patients, respectively. The fact that influenza has a similar global clinical impact on the community to that of RSV infection, but represents a greater socio-economic burden, may contribute to broadening the acceptance of influenza vaccination. PMID- 16216115 TI - Report of the Consensus Conference on Antibiotic Resistance; Prevention and Control (ARPAC). AB - Antimicrobial resistance is a key public health concern in Europe. It is known that there are significant variations in the prevalence of resistance across Europe, and methods to reduce the problem are also assumed to vary significantly. The 'Antibiotic Resistance; Prevention and Control (ARPAC)' Concerted Action project was funded by the European Commission and conducted by four study groups of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID). The project established a network of European hospitals and collated data on antimicrobial resistance prevalence, antimicrobial susceptibility testing methods, typing methods employed, antimicrobial use, antibiotic policies and practices, and infection control policies and practices. The ARPAC Consensus Conference, entitled 'Control of antibiotic resistance in European hospitals informing future evidence-based practice', was held in Amsterdam in November 2004. The conference was co-hosted by the European Commission, ESCMID and the Dutch Working Party on Antibiotic Policy (SWAB). Key ARPAC findings were presented and discussed in the context of the worldwide situation. The conference delivered a set of high-priority recommendations likely to have a significant impact on antimicrobial resistance. This report summarises these recommendations. PMID- 16216114 TI - Metallo-beta-lactamases produced by carbapenem-resistant Pseudomonas aeruginosa in Brazil. PMID- 16216116 TI - Molecular characterisation of the caprine (Capra hircus) lymphocyte function associated antigen-1 alpha subunit-encoding cDNA. AB - BACKGROUND: Lymphocyte function-associated antigen-1 (LFA-1, CD11a/CD18, alpha L beta 2) is required for many cellular adhesive interactions during the immune response. METHODS: We used SMART RACE technology to obtain caprine CD11a 5'- and 3'-ends and RT-PCR to amplify the full-length CDS. RESULTS: The Capra hircus CD11a-encoding cDNA was sequenced and compared with its human, murine, rat, bovine and ovine counterparts. Despite some focal differences, it shares all the main characteristics of its known mammalian homologues. CONCLUSION: Therefore, along with the caprine CD18-encoding cDNA, which has been available for a few months, the sequence data revealed here will allow the Capra hircus LFA-1 expression in vitro as a tool to explore the specificities of inflammation in the caprine species. PMID- 16216117 TI - Unfavourable birth outcomes of the Roma women in the Czech Republic and the potential explanations: a population-based study. AB - BACKGROUND: Data on the health status of the Roma people in Central and Eastern Europe are sparse and the reasons for their poor health are not clear. The objective of this study was to quantify the differences in birth outcomes between Roma and non-Roma mothers in the Czech Republic and to investigate the potential causes of such differences. METHOD: A population-based study recruited 8938 non Roma and 1388 Roma hospitalised singleton births that occurred in two Czech districts (Teplice and Prachatice) between 1995 and 2004. During their stay in hospital, mothers completed a questionnaire on their demographic and socioeconomic characteristics and maternal smoking and alcohol consumption. Data on maternal height and weight and on infants' birth weight and gestational age were taken from hospital records. RESULTS: Birth weight and gestational age of Roma infants was 373 (SE 15) g and 0.92 (0.05) weeks, respectively, lower than in non-Roma infants. Controlling for demographic, socioeconomic and behavioural factors reduced these differences to 133 (18) g and 0.57 (0.06) weeks, respectively (all p-values < 0.001). In terms of binary outcomes, the Roma vs. non-Roma odds ratios were 4.5 (95% CI 3.7-5.4) for low birth weight (< 2500 g), 2.8 (2.2-3.4) for preterm birth (< 37 weeks of gestation), and 2.9 (2.5-3.4) for intrauterine grown retardation (<10th percentile of birth weight for gestational age); controlling for all covariates reduced these odds ratios to 1.7 (1.3-2.2), 1.5 (1.1-2.0) and 1.3 (1.0-1.6), respectively. Maternal education made the largest contribution to the ethnic differences; the role of health behaviours was relatively modest. CONCLUSION: There are striking differences in birth outcomes between Roma and non-Roma mothers. The causes of these differences are complex but largely socioeconomic. PMID- 16216119 TI - Fat intake and composition of fatty acids in serum phospholipids in a randomized, controlled, Mediterranean dietary intervention study on patients with rheumatoid arthritis. AB - BACKGROUND: We have previously reported that rheumatoid arthritis patients, who adopted a modified Cretan Mediterranean diet, obtained a reduction in disease activity and an improvement in physical function and vitality. This shift in diet is likely to result in an altered intake of fatty acids. Therefore, the objective of the present study was to examine the dietary intake of fatty acids, as well as the fatty acid profile in serum phospholipids, during the dietary intervention study presented earlier. RESULTS: From baseline to the end of the study, changes in the reported consumption of various food groups were observed in the Mediterranean diet group. The change in diet resulted in a number of differences between the Mediterranean diet group and the control diet group regarding the fatty acid intake. For instance, a lower ratio of n-6 to n-3 fatty acids was observed in the Mediterranean diet group, both assessed by diet history interviews (dietary intake) and measured in serum phospholipids. Moreover, the patients in the Mediterranean diet group that showed a moderate or better clinical improvement during the study (diet responders), had a higher reported intake of n-3 fatty acids and a lower ratio of n-6 to n-3 fatty acids compared to the patients with minor or no improvement. Also the fatty acid profile in serum phospholipids differed in part between the diet responders and the diet non responders. CONCLUSION: The changes in the fatty acid profile, indicated both by dietary assessments and through fatty acids in s-phospholipids may, at least in part, explain the beneficial effects of the Cretan Mediterranean diet that we have presented earlier. PMID- 16216118 TI - Conservation and co-option in developmental programmes: the importance of homology relationships. AB - One of the surprising insights gained from research in evolutionary developmental biology (evo-devo) is that increasing diversity in body plans and morphology in organisms across animal phyla are not reflected in similarly dramatic changes at the level of gene composition of their genomes. For instance, simplicity at the tissue level of organization often contrasts with a high degree of genetic complexity. Also intriguing is the observation that the coding regions of several genes of invertebrates show high sequence similarity to those in humans. This lack of change (conservation) indicates that evolutionary novelties may arise more frequently through combinatorial processes, such as changes in gene regulation and the recruitment of novel genes into existing regulatory gene networks (co-option), and less often through adaptive evolutionary processes in the coding portions of a gene. As a consequence, it is of great interest to examine whether the widespread conservation of the genetic machinery implies the same developmental function in a last common ancestor, or whether homologous genes acquired new developmental roles in structures of independent phylogenetic origin. To distinguish between these two possibilities one must refer to current concepts of phylogeny reconstruction and carefully investigate homology relationships. Particularly problematic in terms of homology decisions is the use of gene expression patterns of a given structure. In the future, research on more organisms other than the typical model systems will be required since these can provide insights that are not easily obtained from comparisons among only a few distantly related model species. PMID- 16216120 TI - The CD11a partner in Sus scrofa lymphocyte function-associated antigen-1 (LFA-1): mRNA cloning, structure analysis and comparison with mammalian homologues. AB - BACKGROUND: Lymphocyte function-associated antigen-1 (LFA-1, CD11a/CD18, alphaLbeta2), the most abundant and widely expressed beta2-integrin, is required for many cellular adhesive interactions during the immune response. Many studies have shown that LFA-1 is centrally involved in the pathogenesis of several diseases caused by Repeats-in-toxin (RTX)-producing bacteria. RESULTS: The porcine-LFA-1 CD11a (alpha) subunit coding sequence was cloned, sequenced and compared with the available mammalian homologues in this study. Despite some focal differences, it shares all the main characteristics of these latter. Interestingly, as in sheep and humans, an allelic variant with a triplet insertion resulting in an additional Gln-744 was consistently identified, which suggests an allelic polymorphism that might be biologically relevant. CONCLUSION: Together with the pig CD18-encoding cDNA, which has been available for a long time, the sequence data provided here will allow the successful expression of porcine CD11a, thus giving the first opportunity to express the Sus scrofa beta2 integrin LFA-1 in vitro as a tool to examine the specificities of inflammation in the porcine species. PMID- 16216121 TI - Stimulus-dependent effects on tactile spatial acuity. AB - BACKGROUND: Previous studies have shown that spatio-tactile acuity is influenced by the clarity of the cortical response in primary somatosensory cortex (SI). Stimulus characteristics such as frequency, amplitude, and location of tactile stimuli presented to the skin have been shown to have a significant effect on the response in SI. The present study observes the effect of changing stimulus parameters of 25 Hz sinusoidal vertical skin displacement stimulation ("flutter") on a human subject's ability to discriminate between two adjacent or near adjacent skin sites. Based on results obtained from recent neurophysiological studies of the SI response to different conditions of vibrotactile stimulation, we predicted that the addition of 200 Hz vibration to the same site that a two point flutter stimulus was delivered on the skin would improve a subject's spatio tactile acuity over that measured with flutter alone. Additionally, similar neurophysiological studies predict that the presence of either a 25 Hz flutter or 200 Hz vibration stimulus on the unattended hand (on the opposite side of the body from the site of two-point limen testing - the condition of bilateral stimulation - which has been shown to evoke less SI cortical activity than the contralateral-only stimulus condition) would decrease a subject's ability to discriminate between two points on the skin. RESULTS: A Bekesy tracking method was employed to track a subject's ability to discriminate between two-point stimuli delivered to the skin. The distance between the two points of stimulation was varied on a trial-by-trial basis, and several different stimulus conditions were examined: (1) The "control" condition, in which 25 Hz flutter stimuli were delivered simultaneously to the two points on the skin of the attended hand, (2) the "complex" condition, in which a combination of 25 Hz flutter and 200 Hz vibration stimuli were delivered to the two points on the attended hand, and (3) a "bilateral" condition, in which 25 Hz flutter was delivered to the two points on the attended hand and a second stimulus (either flutter or vibration) was delivered to the unattended hand. The two-point limen was reduced (i.e., spatial acuity was improved) under the complex stimulus condition when compared to the control stimulus condition. Specifically, whereas adding vibration to the unilateral two-point flutter stimulus improved spatial acuity by 20 to 25%, the two-point limen was not significantly affected by substantial changes in stimulus amplitude (between 100 - 200 microm). In contrast, simultaneous stimulation of the unattended hand (contralateral to the attended site), impaired spatial acuity by 20% with flutter stimulation and by 30% with vibration stimulation. CONCLUSION: It was found that the addition of 200 Hz vibration to a two-point 25 Hz flutter stimulus significantly improved a subject's ability to discriminate between two points on the skin. Since previous studies showed that 200 Hz vibration preferentially evokes activity in cortical area SII and reduces or inhibits the spatial extent of activity in SI in the same hemisphere, the findings in this paper raise the possibility that although SI activity plays a major role in two-point discrimination on the skin, influences relayed to SI from SII in the same hemisphere may contribute importantly to SI's ability to differentially respond to stimuli applied to closely spaced skin points on the same side of the body midline. PMID- 16216122 TI - Covalent attachment of the plant natural product naringenin to small glass and ceramic beads. AB - BACKGROUND: Natural products have numerous medicinal applications and play important roles in the biology of the organisms that accumulate them. Few methods are currently available for identifying proteins that bind to small molecules, therefore the discovery of cellular targets for natural products with pharmacological activity continues to pose a significant challenge in drug validation. Similarly, the identification of enzymes that participate in the biosynthesis or modification of natural products remains a formidable bottleneck for metabolic engineering. Flavonoids are one large group of natural products with a diverse number of functions in plants and in human health. The coupling of flavonoids to small ceramic and glass beads provides a first step in the development of high-throughput, solid-support base approaches to screen complex libraries to identify proteins that bind natural products. RESULTS: The utilization of small glass and ceramic beads as solid supports for the coupling of small molecules was explored. Initial characterization of the beads indicated uniform and high capacity loading of amino groups. Once the beads were deemed adequate for the linking of small molecules by the coupling of NHS-fluorescein followed by microscopy, chemical hydrolysis and fluorometry, the flavonoid naringenin was modified with 1,4-dibromobutane, followed by the attachment of aminopropyltriethoxysilane. After NMR structural confirmation, the resulting 7-(4 (3-(triethoxysilyl)propylamino)butoxy) naringenin was attached to the ceramic beads. CONCLUSION: Our results demonstrate that ceramic and glass beads provide convenient solid supports for the efficient and facile coupling of small molecules. We succeeded in generating naringenin-coupled ceramic and glass beads. We also developed a convenient series of steps that can be applied for the solid support coupling of other related flavonoids. The availability of solid-support coupled naringenin opens up new opportunities for the identification of flavonoid binding proteins. PMID- 16216123 TI - An inexpensive and rapid diagnostic method of Koi Herpesvirus (KHV) infection by loop-mediated isothermal amplification. AB - BACKGROUND: Koi Herpesvirus (KHV) affects both juvenile and adult common carp and koi, and is especially lethal to fry. The high mortalities caused by the disease have had a negative impact on the international koi trade. Different diagnostic techniques have been used to detect KHV, including: isolation of the virus in cell culture, electron microscopy, several PCR tests, ELISA and in situ hybridisation. All of these methods are time consuming, laborious and require specialised equipment. RESULTS: A rapid field diagnosis of KHV in common and koi carp was developed using loop-mediated isothermal amplification (LAMP). The LAMP reaction rapidly amplified nucleic acid with high specificity and efficiency under isothermal conditions using a simple water bath. Two methods of extracting DNA from host tissue were compared: extraction by boiling and by using a commercial extraction kit. A set of six primers--two inner primers, two outer primers and two loop primers--was designed from a KHV amplicon. The reaction conditions were optimised for detection of KHV in 60 min at 65 degrees C using Bst (Bacillus stearothermophilus) DNA polymerase. When visualised by gel electrophoresis, the products of the KHV LAMP assay appeared as a ladder pattern, with many bands of different sizes from 50 base-pairs (bp) up to the loading well. The KHV LAMP product could also be simply detected visually by adding SYBR Green I to the reaction tube and observing a colour change from orange to green. All samples positive for KHV by visual detection were confirmed positive by gel electrophoresis. The KHV LAMP had the same sensitivity as a standard PCR assay for the detection of KHV. CONCLUSION: This paper describes an accelerated LAMP assay for diagnosis of KHV. The entire procedure took only 90 minutes to produce a result: 15 minutes for DNA extraction; 60 min for the LAMP reaction; 2 min for visual detection using SYBR Green I. The test can be used under field conditions because the only equipment it requires is a water bath. PMID- 16216124 TI - Changes in accessibility and preferences predict children's future fruit and vegetable intake. AB - BACKGROUND: Most children eat fewer fruits and vegetables than recommended. To be able to design effective interventions, understanding the aetiology of the behaviour is important. Accessibility and preferences have shown to be strong correlates of fruit and vegetable intake in several cross-sectional studies. The aim of this study was to identify predictors of future fruit and vegetable intake and to explore longitudinal patterns of interactions between accessibility and preferences. METHODS: Data presented are based on baseline (September 2001) and follow-up (May/June 2002) surveys of 20 control schools in the Norwegian intervention study Fruits and Vegetables Make the Marks. A total of 816 pupils (77%) completed both baseline and follow-up questionnaires. The average age of the sample at baseline was 11.8 years. The research instrument assessing potential predictor variables was guided by Social Cognitive Theory, and included Accessibility at home, Accessibility at school, Modelling, Intention, Preferences, Self-Efficacy and Awareness of the 5-a-day recommendations. Multiple regression analyses were performed. RESULTS: All independent variables (measured at baseline) were significantly correlated to future fruit and vegetable intake (measured at follow-up). When reported fruit and vegetable intake at baseline (past intake) was included in this model, the effect of the other independent variables diminished. Together with past intake, the observed change in the independent variables from baseline to follow-up explained 43% of the variance in the reported intake at follow-up. Past intake remained the strongest predictor, but changes in accessibility at home and at school, as well as changes in preferences for fruits and vegetables, also explained significant amounts of the variance in fruit and vegetable intake at follow-up. In addition, baseline accessibility was found to moderate the relationship between change in preferences and change in intake. CONCLUSION: Change in accessibility and preferences appear to be important predictors of future fruit and vegetable intake among school children. Interventions should focus on strategies to modify these factors. PMID- 16216125 TI - Comorbidity issues in the pharmacological treatment of Pathological Gambling: a critical review. AB - BACKGROUND: Pathological Gambling (PG) is an impulse control disorder often comorbid with other psychopathology, particularly bipolar spectrum disorders, attention deficit/hyperactivity disorder, obsessive-compulsive disorder (OCD) and substance abuse. This paper reviews the published literature on the pharmacological management of PG, highlighting how clinical and subclinical comorbid psychopathology influences the choice of pharmacological treatment. METHODS: Using Medline, the authors reviewed relevant articles published on this topic from 1995 to 2005, focusing on the best-designed studies for inclusion. RESULTS: Much of the literature on PG-treatment presupposes different theories regarding this disorder. Data suggest the utility of differentiating the pharmacotherapy of pathological gamblers in light of their comorbid profile, specifically assessing for comorbid bipolar, ADHD, OCD, and substance abuse disorders. CONCLUSION: Decisions about pharmacological treatment of PG should take into account current and previous comorbid disorders which influence treatment selection. PMID- 16216127 TI - Host microsatellite alleles in malaria predisposition? AB - BACKGROUND: Malaria is a serious, sometimes fatal, disease caused by Plasmodium infection of human red blood cells. The host-parasite co-evolutionary processes are well understood by the association of coding variations such as G6PD, Duffy blood group receptor, HLA, and beta-globin gene variants with malaria resistance. The profound genetic diversity in host is attributed to polymorphic microsatellites loci. The microsatellite alleles in bacterial species are known to have aided their survival in fatal environmental conditions. The fascinating question is whether microsatellites are genomic cushion in the human genome to combat disease stress and has cause-effect relationships with infections. PRESENTATION OF THE HYPOTHESIS: It is hypothesized that repeat units or alleles of microsatellites TH01 and D5S818, located in close proximity to beta-globin gene and immune regulatory region in human play a role in malaria predisposition. Association of alleles at aforesaid microsatellites with malaria infection was analysed. To overrule the false association in unrecognized population stratification, structure analysis and AMOVA were performed among the sampled groups. TESTING OF HYPOTHESIS: Associations of microsatellite alleles with malaria infection were verified using recombination rate, Chi-square, and powerful likelihood tests. Further investigation of population genetic structure, and AMOVA was done to rule out the confounding effects of population stratification in interpretation of association studies. IMPLICATION OF THE HYPOTHESIS: Lower recombination rate (theta) between microsatellites and genes implicated in host fitness; positive association between alleles-13 (D5S818), 9 (TH01) and strong susceptibility to Plasmodium falciparum; and alleles-12 (D5S818) and 6 (TH01) rendering resistance to human host were evident. The interesting fact emerging from the study was that while predisposition to malaria was a prehistoric attribute, among TH01 alleles; evolution of resistant allele-6 was a recent phenomenon, which could conceivably be driven by infection related selective forces. The host's microsatellite allelic associations with malaria infection were valid in the light of low genetic variance between sampled groups and no population stratification. PMID- 16216126 TI - Rhinovirus infection induces cytotoxicity and delays wound healing in bronchial epithelial cells. AB - BACKGROUND: Human rhinoviruses (RV), the most common triggers of acute asthma exacerbations, are considered not cytotoxic to the bronchial epithelium. Recent observations, however, have questioned this knowledge. The aim of this study was to evaluate the ability of RV to induce epithelial cytotoxicity and affect epithelial repair in-vitro. METHODS: Monolayers of BEAS-2B bronchial epithelial cells, seeded at different densities were exposed to RV serotypes 1b, 5, 7, 9, 14, 16. Cytotoxicity was assessed chromatometrically. Epithelial monolayers were mechanically wounded, exposed or not to RV and the repopulation of the damaged area was assessed by image analysis. Finally epithelial cell proliferation was assessed by quantitation of proliferating cell nuclear antigen (PCNA) by flow cytometry. RESULTS: RV1b, RV5, RV7, RV14 and RV16 were able to induce considerable epithelial cytotoxicity, more pronounced in less dense cultures, in a cell-density and dose-dependent manner. RV9 was not cytotoxic. Furthermore, RV infection diminished the self-repair capacity of bronchial epithelial cells and reduced cell proliferation. CONCLUSION: RV-induced epithelial cytotoxicity may become considerable in already compromised epithelium, such as in the case of asthma. The RV-induced impairment on epithelial proliferation and self-repair capacity may contribute to the development of airway remodeling. PMID- 16216128 TI - Hepatocyte and keratinocyte growth factors and their receptors in human lung emphysema. AB - BACKGROUND: Hepatocyte and keratinocyte growth factors are key growth factors in the process of alveolar repair. We hypothesized that excessive alveolar destruction observed in lung emphysema involves impaired expression of hepatocyte and keratinocyte growth factors or their respective receptors, c-met and keratinocyte growth factor receptor. The aim of our study was to compare the expression of hepatocyte and keratinocyte growth factors and their receptors in lung samples from 3 groups of patients: emphysema; smokers without emphysema and non-smokers without emphysema. METHODS: Hepatocyte and keratinocyte growth factor proteins were analysed by immunoassay and western blot; mRNA expression was measured by real time quantitative polymerase chain reaction. RESULTS: Hepatocyte and keratinocyte growth factors, c-met and keratinocyte growth factor receptor mRNA levels were similar in emphysema and non-emphysema patients. Hepatocyte growth factor mRNA correlated negatively with FEV1 and the FEV1/FVC ratio both in emphysema patients and in smokers with or without emphysema. Hepatocyte and keratinocyte growth factor protein concentrations were similar in all patients' groups. CONCLUSION: The expression of hepatocyte and keratinocyte growth factors and their receptors is preserved in patients with lung emphysema as compared to patients without emphysema. Hepatocyte growth factor mRNA correlates with the severity of airflow obstruction in smokers. PMID- 16216129 TI - Extension of Lander-Waterman theory for sequencing filtered DNA libraries. AB - BACKGROUND: The degree to which conventional DNA sequencing techniques will be successful for highly repetitive genomes is unclear. Investigators are therefore considering various filtering methods to select against high-copy sequence in DNA clone libraries. The standard model for random sequencing, Lander-Waterman theory, does not account for two important issues in such libraries, discontinuities and position-based sampling biases (the so-called "edge effect"). We report an extension of the theory for analyzing such configurations. RESULTS: The edge effect cannot be neglected in most cases. Specifically, rates of coverage and gap reduction are appreciably lower than those for conventional libraries, as predicted by standard theory. Performance decreases as read length increases relative to island size. Although opposite of what happens in a conventional library, this apparent paradox is readily explained in terms of the edge effect. The model agrees well with prototype gene-tagging experiments for Zea mays and Sorghum bicolor. Moreover, the associated density function suggests well-defined probabilistic milestones for the number of reads necessary to capture a given fraction of the gene space. An exception for applying standard theory arises if sequence redundancy is less than about 1-fold. Here, evolution of the random quantities is independent of library gaps and edge effects. This observation effectively validates the practice of using standard theory to estimate the genic enrichment of a library based on light shotgun sequencing. CONCLUSION: Coverage performance using a filtered library is significantly lower than that for an equivalent-sized conventional library, suggesting that directed methods may be more critical for the former. The proposed model should be useful for analyzing future projects. PMID- 16216131 TI - The place of public inquiries in shaping New Zealand's national mental health policy 1858-1996. AB - BACKGROUND: This paper discusses the role of public inquiries as an instrument of public policy-making in New Zealand, using mental health as a case study. The main part of the paper analyses the processes and outcomes of five general inquiries into the state of New Zealand's mental health services that were held between 1858 and 1996. RESULTS: The membership, form, style and processes used by public inquiries have all changed over time in line with constitutional and social trends. So has the extent of public participation. The records of five inquiries provide periodic snapshots of a system bedevilled by long-standing problems such as unacceptable standards, under-resourcing, and poor co ordination. Demands for an investigation no less than the reports and recommendations of public inquiries have been the catalyst of some important policy changes, if not immediately, then by creating a climate of opinion that supported later change. Inquiries played a significant role in establishing lunatic asylums, in shaping the structure of mental health legislation, establishing and maintaining a national mental health bureaucracy within the machinery of government, and in paving the way for deinstitutionalisation. Ministers and their departmental advisers have mediated this contribution. CONCLUSION: Public inquiries have helped shape New Zealand's mental health policy, both directly and indirectly, at different stages of evolution. In both its advisory and investigative forms, the public inquiry remains an important tool of public administration. The inquiry/cause and policy/effect relationship is not necessarily immediate but may facilitate changes in public opinion with corresponding policy outcomes long after any direct causal link could be determined. When considered from that long-term perspective, the five inquiries can be linked to several significant and long-term contributions to mental health policy in New Zealand. PMID- 16216130 TI - A confocal microscopic study of solitary pulmonary neuroendocrine cells in human airway epithelium. AB - BACKGROUND: Pulmonary neuroendocrine cells (PNEC) are specialized epithelial cells that are thought to play important roles in lung development and airway function. PNEC occur either singly or in clusters called neuroepithelial bodies. Our aim was to characterize the three dimensional morphology of PNEC, their distribution, and their relationship to the epithelial nerves in whole mounts of adult human bronchi using confocal microscopy. METHODS: Bronchi were resected from non-diseased portions of a lobe of human lung obtained from 8 thoracotomy patients (Table 1) undergoing surgery for the removal of lung tumors. Whole mounts were stained with antibodies to reveal all nerves (PGP 9.5), sensory nerves (calcitonin gene related peptide, CGRP), and PNEC (PGP 9.5, CGRP and gastrin releasing peptide, GRP). The analysis and rendition of the resulting three-dimensional data sets, including side-projections, was performed using NIH Image software. Images were colorized and super-imposed using Adobe Photoshop. RESULTS: PNEC were abundant but not homogenously distributed within the epithelium, with densities ranging from 65/mm2 to denser patches of 250/mm2, depending on the individual wholemount. Rotation of 3-D images revealed a complex morphology; flask-like with the cell body near the basement membrane and a thick stem extending to the lumen. Long processes issued laterally from its base, some lumenal and others with feet-like processes. Calcitonin gene-related peptide (CGRP) was present in about 20% of PNEC, mainly in the processes. CGRP-positive nerves were sparse, with some associated with the apical part of the PNEC. CONCLUSION: Our 3D-data demonstrates that PNEC are numerous and exhibit a heterogeneous peptide content suggesting an active and diverse PNEC population. PMID- 16216133 TI - Physical stature and intelligence as predictors of the timing of baby boomers' very first dates. AB - The paper analyses to what extent the timing of the very first date of American adolescents in the 1960s correlated with their physical stature and cognitive ability. Using an event history approach, it was found that intelligence and a weight slightly above the average raised the odds of arranging a first rendezvous, whereas the effect of height was gender-specific. Tallness improved chances among boys, whereas girls whose height was slightly below the average had the highest odds. This pattern was mirrored by the adolescents' evaluation of their own physical stature. The fact that the estimated associations are remarkably similar to those observed in the literature on fertility and wages suggests that these attributes are selection criteria in situations of social interaction already among adolescents. PMID- 16216134 TI - The tumour necrosis factor receptor-associated periodic syndrome: current concepts. AB - The tumour necrosis factor receptor (TNFR)-associated periodic syndrome (TRAPS) is an autosomal dominant, multisystemic, autoinflammatory disorder caused by mutations in the TNFR1 gene ( TNFRSF1A ). Traps seems to be the most common hereditary periodic fever (HPF) syndrome in some western populations, and the second most prevalent HPF worldwide, behind familial mediterranean fever (FMF). The proteins involved in susceptibility to TRAPS (TNFRSF1A) and FMF (pyrin) are both members of the death-domain-fold superfamily. Mutations affecting these proteins might cause dysregulation of innate immune responses, with a propensity to autoinflammation. Most TRAPS patients have reduced blood levels of soluble TNFRSF1A between attacks, with an inappropriately small increase during bouts of fever. The pathogenesis of the 'hyperinflammatory state' in TRAPS has been variously ascribed to a shedding defect of TNFRSF1A from the cell surface resulting in increased TNF inflammatory signalling, or impaired TNF apoptotic signalling. Some low-penetrance TNFRSF1A variants also contribute to the clinical phenotype in individuals carrying other HPF-associated mutations, and have been reported in several disorders such as Behcet's disease and systemic lupus erythematosus. Synthetic anti-TNF agents provide a rational form of therapy for TRAPS, and have been shown to delay or indeed prevent development of systemic amyloidosis (AA type), a life-threatening complication in this condition. PMID- 16216132 TI - Urodynamic effects of oxybutynin and tolterodine in conscious and anesthetized rats under different cystometrographic conditions. AB - BACKGROUND: Antimuscarinic agents are the most popular treatment for overactive bladder and their efficacy in man is well documented, producing decreased urinary frequency and an increase in bladder capacity. During cystometry in rats, however, the main effect reported after acute treatment with antimuscarinics is a decrease in peak micturition pressure together with little or no effect on bladder capacity. In the present experiments we studied the effects, in rats, of the two most widely used antimuscarinic drugs, namely oxybutynin and tolterodine, utilising several different cystometrographic conditions. The aim was to determine the experimental conditions required to reproduce the clinical pharmacological effects of antimuscarinic agents, as seen in humans, in particular their ability to increase bladder capacity. RESULTS: Intravenous or oral administration of tolterodine or oxybutynin in conscious rats utilized 1 day after catheter implantation and with saline infusion at constant rate of 0.1 ml/min, gave a dose-dependent decrease of micturition pressure (MP) with no significant change in bladder volume capacity (BVC). When the saline infusion rate into the bladder was decreased to 0.025 ml/min, the effect of oral oxybutynin was similar to that obtained with the higher infusion rate. Also, experiments were performed in rats in which bladders were infused with suramin (3 and 10 microM) in order to block the non-adrenergic, non-cholinergic component of bladder contraction. Under these conditions, oral administration of oxybutynin significantly reduced MP (as observed previously), but again BVC was not significantly changed. In conscious rats with bladders infused with diluted acetic acid, both tolterodine and oxybutynin administered at the same doses as in animals infused with saline, reduced MP, although the reduction appeared less marked, with no effect on BVC. In conscious rats utilized 5 days after catheter implantation, a situation where inflammation due to surgery is reduced, the effect of tolterodine (i.v.) and oxybutynin (p.o.) on MP was smaller and similar, respectively, to that observed in rats utilized 1 day after catheter implantation, but the increase of BVC was not statistically significant. In anesthetized rats, i.v. administration of oxybutynin again induced a significant decrease in MP, although it was of questionable relevance. Both BVC and threshold pressure were not significantly reduced. The number and amplitude of high frequency oscillations in MP were unmodified by treatment. Finally, in conscious obstructed rats, intravenous oxybutynin did not modify frequency and amplitude of non-voiding contractions or bladder capacity and micturition volume. CONCLUSION: Despite the different experimental conditions used, the only effect on cystometrographic parameters of oxybutynin and tolterodine in anesthetized and conscious rats was a decrease in MP, whereas BVC was hardly and non-significantly affected. Therefore, it is difficult to reproduce in rats the cystometrographic increase in BVC as observed in humans after chronic administration of antimuscarinic agents, whereas the acute effects seem more similar. PMID- 16216135 TI - Molecular studies on the seasonal occurrence and development of five myxozoans in farmed Salmo trutta L. AB - Five myxozoan species, Tetracapsuloides bryosalmonae, Sphaerospora truttae, Chloromyxum schurovi, Chloromyxum truttae and a Myxobolus species were detected in farmed brown trout, Salmo trutta L. from Central Scotland. Using PCR and in situ hybridization, this study investigated the seasonal occurrence and tissue location of these species in young of the year brown trout. C. schurovi, C. truttae and Myxobolus sp. were first detected in brown trout in April, 2 months before T. bryosalmonae and S. truttae. T. bryosalmonae and S. truttae showed proliferation in the blood with intravascular stages of T. bryosalmonae accumulating in the heart. In contrast, only small amounts of PCR products of C. schurovi and C. truttae were obtained from the blood, suggesting that these species use the vascular system for transport but proliferate only in their target tissues from which large amounts of PCR product were obtained and where parasites were visible in histological sections. Large amounts of PCR product were obtained for T. bryosalmonae, S. truttae and both Chloromyxum species from the gills of brown trout, suggesting the gills as entry locus for these species. The neurotropic Myxobolus species formed plasmodia predominantly in the peripheral nerves, possibly indicating an entry route through the skin. Presporogonic stages of all other species had disappeared by September and mature spores were present from August onwards. PMID- 16216136 TI - Tick-borne Great Island Virus: (I) Identification of seabird host and evidence for co-feeding and viraemic transmission. AB - Great Island Virus (GIV) is an arbovirus present in the tick Ixodes uriae, a common ectoparasite of nesting seabirds. Common guillemot (Uria aalge) and black legged kittiwake (Rissa tridactyla) are the preferred and most abundant hosts of I. uriae on the Isle of May, Scotland. As part of a study to understand the epidemiology of GIV, the ability of guillemot and kittiwake to support tick-borne transmission of GIV was examined. GIV was present in ticks feeding in isolated guillemot colonies and guillemots had virus-specific neutralizing antibodies demonstrating previous GIV infection. By contrast, only uninfected ticks were found in colonies inhabited solely by kittiwakes. GIV was isolated from kittiwake ticks in colonies which also contained breeding guillemots but no virus-specific neutralizing antibodies were present in blood samples of kittiwake on which infected ticks were feeding. Thus guillemots are the main vertebrate hosts of GIV on the Isle of May whereas kittiwakes do not appear to be susceptible to infection. Virus infection of adult ticks feeding on guillemots was highly efficient and may involve both viraemic transmission and transmission from infected to uninfected ticks feeding together on birds that do not develop a patent viraemia. PMID- 16216138 TI - Workers' involvement--a missing component in the implementation of occupational safety and health management systems in enterprises. AB - Effective implementation of occupational safety and health (OSH) legislation based on European Union directives requires promotion of OSH management systems (OSH MS). To this end, voluntary Polish standards (PN-N-18000) have been adopted, setting forth OSH MS specifications and guidelines. However, the number of enterprises implementing OSH MS has increased slowly, falling short of expectations, which call for a new national policy on OSH MS promotion. To develop a national policy in this area, a survey was conducted in 40 enterprises with OSH MS in place. The survey was aimed at identifying motivational factors underlying OSH MS implementation decisions. Specifically, workers' and their representatives' involvement in OSH MS implementation was investigated. The results showed that the level of workers' involvement was relatively low, which may result in a low effectiveness of those systems. The same result also applies to the involvement of workers' representatives and that of trade unions. PMID- 16216137 TI - Lack of immunological cross-reactivity between parasite-derived and recombinant forms of ES-62, a secreted protein of Acanthocheilonema viteae. AB - The longevity of filarial nematodes is dependent on secreted immunomodulatory products. Previous investigation of one such product, ES-62, has suggested a critical role for post-translationally attached phosphorylcholine (PC) moieties. In order to further investigate this, ES-62 lacking PC was produced, using the Pichia pastoris recombinant gene expression system. Unlike parasite-derived ES 62, which is tetrameric the recombinant material was found to consist of a mixture of apparently stable tetramers, dimers and monomers. Nevertheless, the recombinant protein was considered to be an adequate PC-free ES-62 as it was recognized by existing antisera against the parasite-derived protein. However, subsequent to this, recognition of parasite-derived ES-62 by antibodies produced against the recombinant protein was found to be absent. In an attempt to explain this, recombinant ES-62 was subjected to structural analysis and was found to (i) contain 3 changes in amino acid composition; (ii) demonstrate significant alterations in glycosylation; (iii) show major differences in protein secondary structure. The effects of these alterations in relation to the observed change in immunogenicity were investigated and are discussed. The data presented clearly show that recognition by existing antibodies is insufficient proof that recombinant proteins can be used to mimic parasite-derived material in studies on nematode immunology and vaccination. PMID- 16216148 TI - Addiction and the medical profession. PMID- 16216149 TI - Efficacy and tolerability of long-acting injectable naltrexone for alcohol dependence. PMID- 16216150 TI - Recent advances in the psychotherapy of addictive disorders. AB - Rigorous research has recently identified a range of behavioral therapies that have been shown to be effective across the most prevalent types of substance dependence. This review summarizes the roles of behavioral therapies as contrasted with those of pharmacotherapies for substance use disorders and then provides an overview of the major classes of behavioral therapies (clinical management, coping skills approaches, motivational interviewing, and family and interpersonal approaches), highlighting their effectiveness across cocaine, opioid, alcohol, and cannabis use disorders. Lastly, important areas of current research emphasis including combined treatments, strategies for effectively transporting empirically validated treatments into clinical practice, and increasing the efficiency of treatment are described. PMID- 16216151 TI - Nonmedical use and abuse of scheduled medications prescribed for pain, pain related symptoms, and psychiatric disorders: patterns, user characteristics, and management options. AB - The nonmedical use of scheduled medications commonly prescribed for pain, pain related symptoms, and psychiatric disorders began rising in the mid-1990s. Physicians are confronted with the dilemma of how to minimize the abuse and diversion potential of these products without compromising access for patients with a legitimate medical need. Using data from the National Survey on Drug Use and Health, we describe the scope of nonmedical use of opioids, stimulants, and tranquilizers; characteristics of nonmedical users; and options available to reduce abuse liability. In 2003, lifetime prevalence estimates of nonmedical use were 31.2 million for opioids, 20.7 million for stimulants, and 20.2 million for tranquilizers. Nonmedical users of psychotherapeutics were more likely to be Caucasian; use alcohol, cocaine, or heroin; and to use needles to inject drugs than those who reported using illicit drugs only. Sources of diversion are enumerated, and options for minimizing the abuse liability associated with these medications are described. PMID- 16216152 TI - Expanding treatment of tobacco dependence. AB - Nicotine dependence is the leading preventable cause of adult morbidity and mortality in the world. New research on the treatment of this disorder ranges from studies evaluating access to treatment to studies elucidating the molecular mechanisms of nicotine addiction. As our understanding of the neurobiology of tobacco addiction grows, the number of potential therapeutic targets by which we can intervene in this pernicious disorder also increases. This paper presents an overview of recent research trends in the treatment of tobacco dependence. We review several novel mechanisms of action that may serve as therapeutic targets for the pharmacologic treatment of tobacco dependence, including drugs that affect monamine oxidase, selective nicotinic receptors, glutamate and gamma aminobutyric acid receptors, and the endocannabinoid system. For each of these therapeutic targets, we discuss medications in development that affect these pathophysiologic mechanisms. PMID- 16216153 TI - Training psychiatrists to diagnose and treat substance abuse disorders. AB - Addiction training in psychiatric residency programs needs expansion. Epidemiology research has shown that patients with substance use disorders and co occurring mental health disorders are the norm in nearly all clinical settings. Unfortunately, traditional training approaches built around brief rotations on detoxification or intensive substance abuse rehabilitation units do not adequately train psychiatrists in long-term management skills, and may reinforce misperceptions that these patients do not respond to treatment. An enhanced addiction curriculum coupled with an extended outpatient clinic rotation is an ideal model for teaching the skills needed to successfully care for these patients. Training must include an adequate knowledge base, an opportunity to cultivate positive attitudes toward these patients, and recognition that psychiatrists must take responsibility for treating the addiction problem and any co-occurring psychiatric disorders. The program developed at Boston University Medical Center successfully integrates expanded addiction psychiatry training into the general psychiatry residency. PMID- 16216155 TI - Risperidone for autistic disorder. PMID- 16216156 TI - Repetitive transcranial magnetic stimulation after acute ischemic stroke. PMID- 16216154 TI - The marijuana withdrawal syndrome: diagnosis and treatment. AB - A subset of marijuana smokers develop a cannabis use disorder and seek treatment for their marijuana use on their own initiative. A less well-known consequence of daily, repeated marijuana use is a withdrawal syndrome, characterized by a time dependent constellation of symptoms: irritability, anxiety, marijuana craving, decreased quality and quantity of sleep, and decreased food intake. Treatment studies show that rates of continuous abstinence are low (comparable to relapse rates for other abused drugs), and more treatment options are needed. The objective of this review is to update clinicians on the current state of marijuana research and to describe features of marijuana withdrawal to facilitate the diagnosis and treatment of cannabis use disorders. PMID- 16216157 TI - Neuropsychiatric aspects of migraine. AB - Migraine is a common chronic disorder that presents with recurrent attacks of headache and associated symptoms. Various somatic and psychiatric conditions have been reported as comorbid conditions with migraine. Among the psychiatric disorders that have been reported as comorbid with migraine are major depression, anxiety, panic disorder, and bipolar disorder. Recent studies discussing the comorbidity of headache and psychiatric disorders are reviewed here. PMID- 16216158 TI - Treatment approaches to symptoms associated with frontotemporal degeneration. AB - A systematic review of the literature reveals few randomized, controlled trials for drug therapy in frontotemporal degeneration (FTD). Although there is evidence to support a serotonergic deficit, and clinicians frequently prescribe selective serotonin reuptake inhibitors to patients with FTD, only paroxetine and trazodone have been studied. There is Class II evidence for use of rivastigmine in FTD behavioral disturbances, although there is no consistent evidence of cholinergic deficit in this illness. PMID- 16216159 TI - Transcranial magnetic stimulation for the treatment of depression in neurologic disorders. AB - Depression is commonly associated with neurologic disorders. Although depression in neurologic conditions often is associated with a negative impact on quality of life, it frequently is poorly managed. Some factors, such as a multidrug regimen, lack of efficacy, and side effects of antidepressants may explain why depression is not adequately treated in patients with neurologic disorders. Therefore, this population needs new approaches for depression treatment, and repetitive transcranial magnetic stimulation (rTMS) may be one of them because it has been shown to be effective for the treatment of depression alone and depression in certain neurologic diseases such as Parkinson's disease and stroke. rTMS is a noninvasive, focal, and painless treatment associated with few, mild side effects. It may be effective in the treatment of neurologic diseases such as Parkinson's disease, stroke, and epilepsy. In this paper, we discuss the potential risks and benefits of rTMS treatment for depression in Parkinson's disease, epilepsy, stroke, multiple sclerosis, and Alzheimer's disease. Lastly, a framework that includes the parameters of stimulation (intensity, frequency, number of pulses, and site of stimulation) for the treatment of depression in neurologic diseases is proposed. PMID- 16216160 TI - Neurobehavioral effects of interferon therapy. AB - Interferon (IFN) therapy is associated with neuropsychiatric side effects including cognitive dysfunction and mood syndromes of varying severity. These problems are the most common causes of treatment discontinuation. Dose and duration of treatment influence risk of IFN-induced side effects. Rates of IFN induced depression vary, but approach 50% in recent studies. Presence and severity of depressive symptoms at or before treatment predicts development of mood disorders during IFN therapy. Several possible endocrine and neurotransmitter perturbations may be responsible for IFN neurotoxicity, with recent research suggesting different symptoms clusters are related to different underlying mechanisms. The interpretation of these clusters has been influenced by subjective versus objective evaluation of cognitive function. Effective management of IFN-induced neuropsychiatric side effects should involve pretreatment screening and interval assessment during therapy. Antipsychotic and psychostimulant drugs may be used against cognitive dysfunction. Antidepressants have been shown to be effective against IFN-induced depression and can be very valuable in support of adequate or completed therapy. PMID- 16216162 TI - Rhinosinusitis and the allergist. PMID- 16216161 TI - Update on depression in neurologic illness: stroke, epilepsy, and multiple sclerosis. AB - The risk of depression is increased in chronic neurologic illness and can adversely affect the course of disease. Recent literature is reviewed for depression in stroke, epilepsy, and multiple sclerosis. Depression can share pathophysiologic aspects of the comorbid illness, such as neurotransmitter pathway disturbances, hypothalamus-pituitary-adrenal pathway disturbances, and changes in immunologic function. Depression also can be a psychologic reaction to the burden of the neurologic condition. Risk factors for development of depression are reviewed. Depression and other medical conditions can have shared symptoms (eg, fatigue, psychomotor retardation) that can complicate the diagnosis of depression in neurologic illness. Proper selection of antidepressant treatment is necessary to avoid worsening the neurologic disorder. PMID- 16216166 TI - Outcomes in pediatric asthma. AB - Pediatric asthma is one of the most common chronic conditions seen in children throughout the world. Even with our better understanding of the pathology of pediatric asthma and improved pharmaceutical management for this disorder, we are still far from reaching the goals of optimal asthma outcomes as outlined by the National Heart, Lung, and Blood Institute of the National Institutes of Health. What we mean by outcomes is the change in the patient's current and future health due to the care given. The outcomes in pediatric asthma can be divided into three categories: clinical, humanistic, and economic. This article provides information regarding the different components of outcomes in pediatric asthma and how one can use outcomes to improve asthma control for the child. PMID- 16216167 TI - Pediatric food allergy update. AB - Food allergy is mostly a childhood disease that differs from the adult-onset allergy in several aspects, including the prevalence, commonly offending foods, symptomatology, and prognosis. The prevalence is much overestimated by the public, but it probably affects up to 6% of children. In some cases, the diagnosis might be clearly evident, but in most cases an expert evaluation may be needed that usually includes verification by challenge testing. Currently, the management of food allergy is avoidance of the offending foods together with the availability of pharmacologic agents for symptomatic treatment, including self administered epinephrine. The latter is of utmost importance in highly allergic subjects because accidental trivial exposure is not uncommon. Recent studies promise immunomodulatory agents that might be of potential use for prophylaxis. One such agent is anti-immunoglobulin E, which might be available in the near future. PMID- 16216168 TI - The environment and risk factors for atopy. AB - Atopy is the genetic potential to manifest the trinity of classic allergic diseases--atopic dermatitis, allergic rhinitis, and asthma. All have been observed to have increased over the past several decades. The interplay of the genetics of atopy with various environmental exposures may manifest itself in the form of allergic phenotypes or paradoxically may actually suppress the allergic response. The hygiene hypothesis offers an explanation of why certain environmental exposures early in life may suppress or activate clinical disease. Primary prevention is directed at preventing the clinical manifestations of atopy by suppressing or delaying the onset of allergic sensitivity. Studies of primary prevention have brought mixed results. Secondary prevention is directed at reducing or removing triggers in the environment of the sensitized individual. Secondary prevention measures directed at one allergen have not proven successful. Comprehensive intervention programs dealing with both allergens and other potential triggers appear beneficial. PMID- 16216169 TI - Ethical considerations in research involving children. AB - Ethical concerns and medical advances unique to children and adolescents make pediatric research an evolving endeavor. The child's changing physiology from infancy to maturity requires understanding of the benefits and risks of medical therapies currently available, but often not tested in children, as well as the risks and benefits of inclusion of children in clinical research. Recent mandates from the US Food and Drug Administration (FDA) require that drugs be tested for safety and efficacy in children. Growing concern for the prevalence of chronic disorders and increasing understanding of distinct phenotypes of these disorders points to a likely need for unique interventions matched to specific phenotypes. All contribute to the increasing complexity of ethical considerations in pediatric research. PMID- 16216172 TI - The role of ubiquitous airborne fungi in chronic rhinosinusitis. AB - Chronic rhinosinusitis (CRS) is a confusing disease for both allergists and otorhinolaryngologists, partially due to its poorly understood pathophysiology and partially due to its limited treatment options. Several recent reports now provide evidence for a better understanding of the etiology and the relationship of CRS to airborne fungi, especially to Alternaria. First, the development of novel methods enables detection of certain fungi in mucus from the nasal and paranasal sinus cavities. Second, a non-immunoglobulin E-mediated immunologic mechanism for reactivity of CRS patients to certain common fungi has been described. Third, these fungi are surrounded by eosinophils in vivo, suggesting that they are targeted by eosinophils. Fourth, the preliminary results of studies using antifungal agents to treat patients with CRS are promising. Overall, these recent discoveries provide a logical mechanism for the pathophysiology of CRS, and they also suggest promising avenues for treatment of CRS with antifungal agents. PMID- 16216171 TI - Pathogenesis of nasal polyps: an update. AB - The cause of nasal polyp formation is still unknown. Genetic predisposition has been suggested, but there are scanty data to support such theories. Activated epithelial cells may be the major source of mediators inducing influx of inflammatory cells (mostly eosinophils) and proliferation and activation of fibroblasts leading to nasal polyp formation. Infectious agents (including viruses, bacteria, or fungi) may be potential primary factors activating nasal epithelial cells. Proinflammatory cytokines and growth factors play important roles in the persistence of mucosal inflammation associated with nasal polyps. Arachidonic acid metabolites seem to be particularly important in the pathogenesis of nasal polyps in patients with aspirin hypersensitivity rhinosinusitis/asthma syndrome. PMID- 16216173 TI - Viral infections and susceptibility to recurrent sinusitis. AB - Viral respiratory infection is thought to be the major contributing factor in the pathogenesis of bacterial sinusitis. It is still not clear why some patients suffer from recurrent sinusitis episodes, as information on the exact pathomechanism of how bacterial sinusitis develops as a complication of viral colds is still scarce. In this review, different mechanisms of how a viral infection may predispose some patients to recurrent bacterial sinusitis episodes are presented. Awareness of the fact that different mechanisms may exist behind this single diagnosis of recurrent sinusitis may be of help when diagnostic measures and treatments are planned for these patients. PMID- 16216174 TI - Chronic sinusitis in children and adults: role of bacteria and antimicrobial management. AB - The nasopharynx serves as the reservoir for anaerobic bacteria as well as pathogenic bacteria that can cause respiratory infections including sinusitis. Some of these organisms possess the ability to interfere with the growth of potential pathogens and may play a role in preventing infections. Anaerobic bacteria emerge as pathogens as the infection becomes chronic. This may result from the selective pressure of antimicrobial agents that enable resistant anaerobic organisms to survive, and from the development of conditions appropriate for anaerobic growth, which include the reduction in oxygen tension and an increase in acidity within the sinus. Anaerobes were identified in chronic sinusitis in adults and children whenever techniques for their cultivation were employed. The predominant isolates were pigmented Prevotella, Fusobacterium, and Peptostreptococcus spp. The choice of antimicrobial agent in chronic sinusitis should provide coverage for the usual pathogens in acute sinusitis as well as beta-lactamase-producing aerobic and anaerobic organisms. PMID- 16216175 TI - Treatment of chronic rhinosinusitis with low-dose, long-term macrolide antibiotics: an evolving paradigm. AB - The 14-membered and 15-membered ring macrolide antibiotics express immunomodulatory effects in chronic respiratory disorders in humans that are distinct from their antimicrobial properties. These drugs downregulate the excessive immune and inflammatory responses observed in these conditions while promoting tissue repair. To this end, chronic rhinosinusitis is characterized by mucosal inflammation of nasal and sinus mucosa for more than 3 months and accounts for significant health care resource allocation due to difficulties in treatment. Clinical efficacy of macrolide antibiotics as biologic response modifiers in patients with chronic rhinosinusitis is suggested by compelling basic research and small, uncontrolled clinical studies. Hence, long-term, prospective double-blind placebo-controlled clinical studies are indicated to establish the utility of these drugs in the treatment of patients with chronic rhinosinusitis. PMID- 16216176 TI - Sinusitis in HIV: microbiology and therapy. AB - Rhinosinusitis in the HIV-infected population is an increasingly common finding as HIV infection has transitioned toward becoming a chronic medical condition. In this patient population, rhinosinusitis may be challenging to diagnose and treat effectively. However, adequate diagnostic tools are available, microbial identity can be reasonably anticipated based on the CD4 count, and effective management strategies can be implemented. In this article, we discuss the diagnostic and therapeutic options for HIV-infected patients with rhinosinusitis. PMID- 16216177 TI - Compartment syndrome: time from diagnosis to fasciotomy. AB - A definitive safe time to fasciotomy for compartment syndrome has not been established. Therefore, the records of 28 patients who had a fasciotomy for compartment syndrome at two trauma centers (18 level I, 10 level II) were reviewed to determine time from diagnosis to fasciotomy and clinical outcome. Average times at the two trauma centers (level I: 160 minutes, range, 50-315 minutes; level II: 105 minutes, range, 51-185 minutes) were significantly different. Ten patients (5 level I, 5 level II) with an average time from diagnosis to fasciotomy of 122 minutes (range, 70-185 minutes) sustained residual deficits. There was no correlation between time from diagnosis to fasciotomy and residual deficits. A time from diagnosis to fasciotomy as short as 70 minutes was associated with residual deficit, but a time as long as 315 minutes (patient with deficits) was associated with no functional deficits. PMID- 16216178 TI - Acetabular dysplasia following closed reduction of developmental dislocation of the hip. AB - The purpose of this study was to determine the incidence of acetabular dysplasia following closed reduction of developmental dislocation of the hip, particularly as it relates to the age at reduction and the presence of the ossific nucleus. Thirty-five children with unilateral developmental dislocation of the hip underwent closed reduction. Medical records were reviewed for gender, side, age at reduction, use of Pavlik harness, prereduction traction, presence/absence of the ossific nucleus, and whether adductor longus tenotomy was performed. Acetabular index was measured. The mean age at reduction was 10 months. The mean length of follow-up was 91 months. Following closed reduction, the incidence of acetabular dysplasia was 69% (24 hips). No statistically significant relationship between acetabular dysplasia and age at reduction or presence of the ossific nucleus was demonstrated. PMID- 16216179 TI - Osteoblastoma of the scaphoid: a case report. AB - Osteoblastoma is an uncommon primary bone tumor that rarely is found in the hand or wrist. Recurrent osteoblastomas often have a more aggressive appearance than the original tumor, and differential diagnosis from osteosarcoma is difficult. The pain that can accompany this tumor is debilitating. Therefore, successful treatment requires complete removal of the tumor. The purpose of this report is to present an unusual case of osteoblastoma of the carpal scaphoid. This tumor was treated successfully by curettage and bone grafting. At 1 year postoperatively, the patient presented with a stable, painless wrist with full range of motion. PMID- 16216180 TI - Neurovascular compression following isolated popliteus muscle rupture: a case report. AB - This case report concerns an unusual complication of neurovascular compression following an isolated popliteus muscle rupture. A 59-year-old man, after a fall from a horse, gradually developed symptoms of a swollen leg, dysesthesias in the sole of his foot, and muscle weakness of his toe flexors. At presentation, he was found to have a complete tibial nerve injury at the level of the popliteal fossa and significant neuropathic pain. MRI demonstrated a rupture in the muscular portion of the popliteus muscle with extensive edema and hemorrhage compressing the tibial nerve in the popliteal fossa. The edema extended up to the distal part of sciatic nerve where there was evidence of intraneural hemorrhage. In the course of recovery, the patient additionally developed deep venous thrombosis in the ipsilateral popliteal vein. Spontaneous recovery was documented on serial clinical and electrodiagnostic examinations. The patient's neuropathic pain improved significantly within 6 months and his neurologic function recovered nearly fully by 2 years. PMID- 16216181 TI - Trans-scaphoid, Transcapitate, Transhamate Injury: A Case Report. AB - Traumatic axial dislocation of the carpus in a 20-year-old man is described. This injury was accompanied by a crushing injury to the hand. The disruption pattern was different from those of previously reported cases. Despite the restoration of painless wrist motion postoperatively, grip strength remained below normal. Early accurate reduction, fixation, and range of motion exercise are the treatments of choice in such complex injuries. PMID- 16216182 TI - Intrapartum coccygeal fracture, a cause for postpartum coccydynia: a case report. AB - Coccydynia can result from a varying number of causes, parturition being one of them. Although strains and sprains of the ligaments attached to the coccyx have been thought to be the usual cause for coccydynia occurring after childbirth, an intrapartum coccygeal fracture dislocation can result in the same. A 28-year-old female presented to the orthopaedic department 4 weeks after the birth of her first child with the complaint of coccygeal pain. Examination revealed marked local tenderness over the coccyx but no crepitus was felt. Radiographs established the diagnosis of fracture and posterior dislocation between the second and third coccygeal fragments. Conservative treatment in the form of rest, doughnut ring, local heat, and avoidance of direct pressure over the area resulted in considerable improvement over the next 4 weeks. Coccygeal fracture dislocation may result in introital dyspareunia and tension myalgia of the pelvic floor. Pain from this lesion may become recurrently symptomatic. The diagnosis must be established at the outset and appropriate treatment instituted to avoid these complications. PMID- 16216183 TI - [Factors influencing incipient diabetic nephropathy: ESODIAH study]. AB - BACKGROUND AND OBJECTIVE: High cholesterol levels might contribute to the presence of albuminuria. The objective of our study was to evaluate the influence of lipid levels on the development of incipient diabetic nephropathy. Secondary objectives were to evaluate the effects of diabetes control, high blood pressure, age, sex, years of diabetes evolution, body mass index and smoking. PATIENTS AND METHOD: 930 subjects were enrolled in an open observational prospective cohort study of subjects with type 2 diabetes mellitus and high cholesterol levels (ESODIAH study) for 2 years. In our nephropathy study we selected 590 patients who had albuminuria measurements done. In every 4-month interval visit we made a clinical evaluation and blood analysis including HbA1c, lipid profile and microalbuminuria. Statistical analysis included t-Student, chi2 test, and binary logistic regressions. RESULTS: 51.7% men, aged 62.08 years of age and with an evolution of their diabetes of 8.49 years were studied. 40.6% had microalbuminuria and 59.4% had normoalbuminuria. High HbA1c correlated with the presence of albuminuria (odds ratio [OR] = 1.3; 95% confidence interval [CI], 1.12-1.55; p = 0.001). The development of microalbuminuria was more frequent in younger (OR = 0.93; 95% CI, 0.89-0.98), smoker (OR = 3.19; 95% CI, 1.02-9.96), subjects with high systolic blood pressure (OR = 1.02; 95% CI, 1-1.05). Total cholesterol levels at the end of the study were higher in new microalbuminuric (group I) than normoalbuminuric patients (group II) (group I: 211.08 [34.75] mg/dl vs group II: 200.67 [30.50]; p = 0.042). CONCLUSIONS: Tobacco, blood pressure and diabetes control influences the presence and development of microalbuminuria. More studies are required to study the influence of hypercholesterolemia. PMID- 16216184 TI - [Glucose intolerance in obese children and adolescents]. AB - BACKGROUND AND OBJECTIVE: The rise in the prevalence of glucose intolerance and type 2 diabetes mellitus in childhood and adolescence in recent decades appears to be closely related to the increase in the incidence of obesity in developed countries. We decided to establish the frequency of glucose intolerance and type 2 diabetes mellitus in a population of obese children and adolescents evaluated at our hospital. PATIENTS AND METHOD: Prospective study of 145 obese patients (60 boys: BMI 29.5 (4.9), BMI z score 4.4 (1.7); and 85 girls: BMI 28.8 (4.6), BMI z score 3.8 (1.4); age range: (4-18 years) who underwent an oral glucose tolerance test (OGTT) between 1998 and 2003. OGTT results were evaluated according to WHO criteria. Insulin secretion and sensitivity parameters (HOMA, QUICKI, area under the curve for glycemia, area under the curve for insulin and insulinogenic index) were also calculated. RESULTS: The frequency of glucose intolerance in the whole population was 19.2%. However, this prevalence varied with age and maturation stage (prepuberty 7.0%, puberty 28.2% and postpuberty 26.5%), and with the obesity degree (BMI z-score between +2 and +3: 8.9%; between +3 and +4: 21.9% and higher than +4: 25%). No type 2 diabetes mellitus cases were observed. CONCLUSIONS: Obese children and adolescents display an elevated incidence of glucose intolerance which seems to be related to the degree of adiposity. PMID- 16216185 TI - [Immunity to diphtheria among adults older than 65 years]. AB - BACKGROUND AND OBJECTIVE: To know the immune state against diphtheria through a survey, a consultation of the vaccination documents and a seroepidemiologic study of an old population. PATIENTS AND METHOD: The immunization status and the prevalence of antidiphtheria antibodies were determined in a representative sample (n = 231) of this population group (> 65 years). Concentrations of toxin antibodies were measured using a commercial ELISA kit. Samples with titres > 0.1 UI/ml were considered protective. RESULTS: In the vaccination documents we only found 19 old individuals vaccinated against tetanus-diphtheria. 47.1% had no protective titres. Geometric mean titres were 0.12 UI/ml. There was no statistically significant difference between the variables age and sex and the prevalence of antibodies. 84.4% of the 122 seropositives did not report having received diphtheria vaccine and neither had documents about the vaccination. CONCLUSION: There were almost no documents about diphtheria vaccination. Therefore, the serologic study is the only reliable way to know the immunity status against diphtheria in elderly people. Approximately 50% of this population need primary immunization. The rest of this group need to have completed antidiphtheria vaccination or a dose booster within a period of 2 years in order to guarantee protective antibody levels against diphtheria. PMID- 16216186 TI - [Effectiveness and safety of calcium antagonists in hypertensive patients]. PMID- 16216187 TI - [Hypercholesterolemia and diabetic nephropathy: a multifactorial association]. PMID- 16216188 TI - [Anaphylaxis to hymenoptera sting: study of 113 patients]. AB - BACKGROUND AND OBJECTIVE: The purpose of this study was to describe the clinical and epidemiological features of anaphylactic reactions to hymenoptera stings, with a case-history analysis according to severity. PATIENTS AND METHOD: We conducted an observational descriptive study of patients aged between 10 and 80 years who suffered a systemic reaction after hymenoptera sting. All of them showed specific serum IgE to venoms from Apis, Vespula and/or Polistes. A questionnaire including history of atopy, past reactions and characteristics of the reaction, was performed by individual interview. Anaphylactic reactions were classified into two levels of severity according to Muller's classification. An analysis of independence was carried out in order to relate each level with several factors: age, gender, atopy, type of previous reactions, area of sting and time sequence. RESULTS: 113 patients were included (63 male; mean age [standard deviation]: 40.1 [15.9] years). Reactions were accounted for bee venom in 10.6% of patients, and wasp in 89.4%. Specific IgE was positive to Vespula in 91.9% of subjects, Polistes in 71.4%, and Apis in 28.7%. Furthermore, 50.4% were sensitive to both Vespula and Polistes. Personal history of atopy was found in 20.3%. Among the 106 patients who reminded previous stings, local large reactions were referred by 35.9% and systemic reactions by 16.5%. Upper limb was the most frequent area of sting (38.9%). Most common symptoms were: pruritus (77.8%), hives (57.5%), edema (54.8%), erythema (52.2%), dizziness (51.3%) and dyspnea (49.5%). Severe reactions occurred in 65.5% of patients. Age, gender, atopy, type of previous reactions, area of sting and restoration time were not significantly associated with severity. Time elapsed to first symptom was proportionally shorter in severe cases (p < 0.05). CONCLUSIONS: There is a high frequency of hypersensitivity to wasp venom (Vespula) in the studied population. Except for immediacy, severity-associated data could not be established. PMID- 16216189 TI - [Therapeutic guidelines. What can be expected from them?]. PMID- 16216190 TI - [Usher syndrome: an example of genetic heterogeneity]. AB - Usher syndrome includes hereditary pathologies characterized by bilateral sensorineural deafness and visual impairment due to retinitis pigmentosa. Clinically, there are three distinct subtypes referred to as USH1, USH2, and USH3. Each subtype is genetically heterogeneous. Eleven different genes are implicated in the pathology; most of them are also implicated in isolated auditory or visual pathologies. MYO7A is responsible of 75% of the USH1 cases and Usherin is responsible of 82% of USH2A patients. The proteins have direct interactions with each other, are expressed in cochlea and retina and perform an essential role in stereocilia homeostasis. From 1995 we approach the study of Usher syndrome in Spain from different points of view: epidemiological, clinic, genetic and molecular. This study will provide additional insight into the pathogenic process involved in Usher syndrome, prognosis factors, and guide to the search for targeted therapies. PMID- 16216191 TI - [A 72 year-old man with chronic rhinosinusal lesions]. PMID- 16216192 TI - [Acute poisoning and chronic exposure to cocaine in a child]. PMID- 16216193 TI - [Guidelines for the appropriate use of do-not-resuscitate orders]. PMID- 16216195 TI - [Use of tables for cardiovascular risk assessment in elderly people]. PMID- 16216196 TI - [Disseminated cutaneous infection by Mycobacterium chelonae after botulinic toxin injection in an immunosuppressed patient]. PMID- 16216198 TI - [Alcoholism in the general hospital: 4 years mortality and hospitalization]. AB - BACKGROUND AND OBJECTIVE: We aimed to analyze the mortality, and emergency room and hospital admissions in a sample of alcohol dependent patients during the four years following their identification in a general hospital. PATIENTS AND METHOD: 198 inpatients from a general hospital who where identified as alcohol dependent and referred during 1998 to the liaison-psychiatry service. RESULTS: 30.8% had died, at a mean age of 57.5 (11.3). Deaths had a direct relationship to the illness responsible for the initial admission to hospital. 42.6% of patients had previous admissions, and 74% had been previously attended at emergency room departments. During the 4 years of follow up, there were new hospital admissions (54.6%), emergency room visits (72.7%) and repeated liaison-psychiatry consultations (20%). Previous admissions to hospital predicted future admissions. Acceptance of alcohol treatment did not reduce the mortality rates. Mortality correlated with age, it was increased in patients with cognitive deficits and was reduced in those with psychiatric co-morbidity. CONCLUSIONS: Late detection of alcohol dependence leads to pour outcomes both in the treatment of alcohol dependence and the concomitant somatic diseases. Early detection strategies of hazardous and harmful alcohol consumption should be implemented in hospital settings. PMID- 16216199 TI - [Determinants of postprandial lipemia measured as diurnal triglyceride profile in non diabetic normolipidemic subjects]. AB - BACKGROUND AND OBJECTIVE: We decided to evaluate the clinical and biochemical predictors of postprandial lipemia, measured as daylong capillarly triglycerides (TGc) profiles, in normolipidemic non diabetic subjects. PATIENTS AND METHOD: We studied 76 normolipidemic non diabetic subjects (45 premenopausal females). Accutrend was used to measure daylong TGc profiles during 3 days in 6 previously standardized points: fasting, pre and 3 h after dinner and lunch and at bedtime. The area under the curve of TGc (AUC-TGc) was determined as expression of postprandial lipemia. RESULTS: Males showed significantly higher AUC-TGc (26.20 [11.00] vs 19.12 [6.57] in females; p < 0.001). Obese showed significantly higher values of AUC-TGc (27.87 [12.47] vs 20.05 [7.04]; p < 0.01). The AUC-TGc correlated with: age (r = 0.242; p < 0.05), body mass index (r = 0.312; p < 0.01), waist circumference (r = 0.394; p < 0.01), fasting plasma triglyceride (r = 0.634; p < 0.001), fasting insulinemia (r = 0.485; p < 0.001) and fasting HOMA (r = 0.484; p < 0.001). The multivariate analysis showed that HOMA (regression coefficient: 0.352; p = 0.02) and waist circumference (regression coefficient: 0.4; p = 0.05) were independent predictors of the AUC-TGc. CONCLUSIONS: Independent determinants of postprandial lipemia were waist circumference and HOMA. PMID- 16216200 TI - [Utility of MALT (Munchner Alkoholismus Test) questionnaire for detection of alcoholism in women]. AB - BACKGROUND AND OBJECTIVE: Patients identified as risk drinkers usually fill a questionnaire in order to assess their alcohol dependence. In our setting, the Munchner Alkoholismus Test (MALT) has been recommended, but there are not enough data about its validity and accuracy, particularly in women. The aim of this study was to assess the psychometric properties of this test in a female population. SUBJECTS AND METHOD: 414 woman were recruited in a Health Center and a Specialized Center for addiction treatment. MALT test and the Schedule for Clinical Assessment in Neuropsychiatry (SCAN) semistructured interview (gold standard; DSM-IV and CIE-10 criteria) were performed to all patients. Internal consistence, criteria validity and the most diagnostic efficacy cut-off point (ROC curves) were assessed. RESULTS: The MALT average (standard deviation) score of the whole sample was 3.98 (9.48) points (95% confidence interval [CI], 3.07 4.90). Cronbach alpha was 0.939 (95% CI, 0.931-0.947). With a score of 4 or more points, the test had 100% (95% CI, 98.9-100) sensitivity and 95.4% (95% CI, 95.2 95.5) specificity. CONCLUSIONS: The MALT test has an excellent internal reliability and extremely good diagnostic accuracy. It is very useful to detect alcohol dependence in women. A cut-off of 4 or more points is proposed. PMID- 16216201 TI - [Alcoholism-related mortality in a general hospital]. PMID- 16216202 TI - [Postprandial lipidemia and obesity]. PMID- 16216203 TI - [Prevalence of obesity in Spain]. PMID- 16216204 TI - [Dermatomycosis]. PMID- 16216205 TI - [Bovine tuberculosis in Spain]. PMID- 16216206 TI - [CD44v6 expression in er-negative and PgR-negative infiltrating ductal breast carcinomas]. PMID- 16216207 TI - [Delirium on hospitalized aged hip fracture patients]. PMID- 16216208 TI - [University of Health Sciences: a blow of hope]. PMID- 16216209 TI - [University of Health Sciences]. PMID- 16216211 TI - [Pachymeningitis as a first manifestation of an overlap vasculitic syndrome]. PMID- 16216213 TI - Clinico-toxinological characterization of the acute effects of the venom of the marine snail, Conus loroisii. AB - The venom of the marine snail, Conus loroisii, was studied to assess its risk and lethal factors in regard of human welfare. The lethality of the crude venom (LD50 5.0 mg/kg via i.p.) in mice was associated with reduced motor activity, asphyxiation, followed by respiratory failure. The effects on vital tissues revealed vascular congestion and inflammatory cell infiltration around the portal triad of the liver, spongiosis of the brain, hemorrhages/congested blood vessels in lung and endothelial cells of the renal tubule. Repeated measures of hematological profiles indicated that the venom significantly reduced erythrocytes (P<0.001, GLM repeated measures), followed associated with depletion of hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin and platelet count. Serum enzymes such as, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, lactate dehydrogenase and alkaline and acid phosphatases were altered significantly (P<0.05, Friedman test), which in turn confirmed the damage of vital organ tissues. Dual effect of the venom on the activity of mouse brain acetylcholinesterase stand for concentration specific, whereas maximal inhibition (60.41%, P<0.05, Wilcoxon signed rank test) in erythrocyte acetylcholinesterase did not show the dual activity observed in brain. The Ciphergen ProteinChip analysis of the envenomed serum further revealed that the venom causes changes in definite molecules involved in inflammatory process and ionic transport. In all, the venom of C. loroisii is potentially lethal to mammals, through its rapid action on the central and peripheral nervous systems by blocking neurotransmission with selective interference of ionic channels/receptors. PMID- 16216214 TI - Modification of recombinatorial cloning for small affinity tag fusion protein construct generation. PMID- 16216215 TI - [How to prevent recurrent nerve palsy during thyroid surgery?]. AB - Recurrent laryngeal nerve (RLN) injury and hypoparathyroidism can occur after thyroid surgery. The rate of RLN injury, mostly transient, ranges from 0.5% to 5% of patients. The risk is more important in patients who undergo reoperative thyroid surgery and in patients with thyroid cancer or hyperthyroidism. Rationales for technique of thyroidectomy are discussed. Meticulous and reproductive surgical technique can lower the postoperative morbidity. However, the potential for RLN injury still exists and must be explained to the patients who are candidate for thyroid surgery. The fact that this information has been delivered during the preoperative visit must be written by the surgeon in the patient's chart. PMID- 16216216 TI - Lysolipid incorporation in dipalmitoylphosphatidylcholine bilayer membranes enhances the ion permeability and drug release rates at the membrane phase transition. AB - The enhanced permeability of lipid bilayer membranes at their gel-to-liquid phase transition has been explained using a "bilayer lipid heterogeneity" model, postulating leaky interfacial regions between still solid and melting liquid phases. The addition of lysolipid to dipalmitoylphosphatidylcholine bilayers dramatically enhances the amount of, and speed at which, encapsulated markers or drugs are released at this, already leaky, phase transition through these interfacial regions. To characterize and attempt to determine the mechanism behind lysolipid-generated permeability enhancement, dithionite permeability and doxorubicin release were measured for lysolipid and non-lysolipid, containing membranes. Rapid release of contents from lysolipid-containing membranes appears to occur through lysolipid-stabilized pores rather than a simple enhancement due to increased drug solubility in the bilayer. A dramatic enhancement in the permeability rate constant begins about two degrees below the calorimetric peak of the thermal transition, and extends several degrees past it. The maximum permeability rate constant coincides exactly with this calorimetric peak. Although some lysolipid desorption from liquid state membranes cannot be dismissed, dialyzation above T(m) and mass spectrometry analysis indicate lysolipid must, and can, remain in the membrane for the permeability enhancement, presumably as lysolipid stabilized pores in the grain boundary regions of the partially melted solid phase. PMID- 16216217 TI - Pharmacology of gap junctions. New pharmacological targets for treatment of arrhythmia, seizure and cancer? AB - Intercellular communication in many organs is maintained via intercellular gap junction channels composed of connexins, a large protein family with a number of isoforms. This gap junction intercellular communication (GJIC) allows the propagation of action potentials (e.g., in brain, heart), and the transfer of small molecules which may regulate cell growth, differentiation and function. The latter has been shown to be involved in cancer growth: reduced GJIC often is associated with increased tumor growth or with de-differentiation processes. Disturbances of GJIC in the heart can cause arrhythmia, while in brain electrical activity during seizures seems to be propagated via gap junction channels. Many diseases or pathophysiological conditions seem to be associated with alterations of gap junction protein expression. Thus, depending on the target disease opening or closure of gap junctions may be of interest, or alteration of connexin expression. GJIC can be affected acutely by changing gap junction conductance or- more chronic--by altering connexin expression and membrane localisation. This review gives an overview on drugs affecting GJIC. PMID- 16216218 TI - Novel aspects of osteoclast activation and osteoblast inhibition in myeloma bone disease. AB - Increased bone resorption is a major characteristic of multiple myeloma and is caused by osteoclast activation and osteoblast inhibition (uncoupling). Myeloma cells alter the local regulation of bone metabolism by increasing the receptor activator of NF-kappaB ligand (RANKL) and decreasing osteoprotegerin expression within the bone marrow microenvironment, thereby stimulating the central pathway for osteoclast formation and activation. In addition, they produce the chemokines MIP-1alpha, MIP-1beta, and SDF-1alpha, which also increase osteoclast activity. On the other hand, myeloma cells suppress osteoblast function by the secretion of osteoblast inhibiting factors, e.g., the Wnt inhibitors DKK-1 and sFRP-2. Moreover, they inhibit differentiation of osteoblast precursors and induce apoptosis in osteoblasts. The resulting bone destruction releases several cytokines, which in turn promote myeloma cell growth. Therefore, the inhibition of bone resorption could stop this vicious circle and not only decrease myeloma bone disease, but also the tumor progression. PMID- 16216219 TI - Betaine improved restriction digestion. AB - Here we report that supplementation of a common compound betaine (1-carboxy-N,N,N trimethylmethanaminium inner salt) enhances restriction digestion of DNA molecules being resistant to digestion despite the existence of recognition sites. A previous study reported total isostabilization of DNA was achieved in the presence of 5.2M of betaine, however, we have observed the enhancement of restriction kinetics at 0.3M of betaine, therefore, it likely provided some catalytic proficiency to restriction enzymes rather than the induction of DNA conformational changes. Betaine also enhances catalytic efficiency of PCR, and our result of restriction digestion, taken together, suggests potential application of betaine in other enzymatic reactions in an aqueous solution. PMID- 16216220 TI - Glucose effect on the expression of 150 kDa oxygen-regulated protein in HeLa cells. AB - Chaperones assist in the correct folding of newly synthesised proteins in the endoplasmic reticulum (ER) of cells, this being essential for the translocation of protein molecules to specific subcellular compartments, extracellular matrix or to biological fluids. The biosynthesis of some ER chaperones is regulated by glucose. They are named "glucose-regulated proteins" (GRPs). The function of some GRPs depends on oxygen, a subgroup named "oxygen-regulated proteins" (ORPs). The biosynthesis of ORPs is induced by deprivation of glucose or oxygen. Exposure of HeLa cells to glucose starvation induces the biosynthesis of various GRPs including ORP 150. The expression of ORP 150 is regulated by the concentration of glucose in the culture medium, being induced by a shortage and repressed by a presence of glucose. We have shown that both glucose starvation and transfection of cells with siRNA (specific to ORP 150 mRNA) evoke similar, but quantitatively different, effects. The cells grown for 72 h in a 4.5 mg/ml glucose-containing medium demonstrated low apoptosis (3.7%) whereas in a 0.5 mg/ml glucose containing medium the apoptosis was increased to 10%. The effect of transfection on apoptosis was distinctly higher with almost 22% of apoptotic cells detected in 72 h cultures. One may conclude that ORP 150 reduces the pro-apoptotic effects of glucose starvation. Such a hypothesis is supported by the observation that the transfection procedure makes HeLa cells resistant to the regulatory effect of glucose on ORP 150 production. The transfected cells do not respond to glucose starvation with an overexpression of ORP 150. It is apparent from our experiments that ORP 150 plays an important role in adaptation of cells to the shortage of glucose and reduces the pro-apoptotic effect of glucose starvation. PMID- 16216221 TI - Expression and characterization of human malaria parasite Plasmodium falciparum origin recognition complex subunit 1. AB - In eukaryotes, the origin recognition complex (ORC) is essential for the initiation of DNA replication. The largest subunit of this complex (ORC1) has a regulatory role in origin activation. Here we report the cloning and functional characterization of Plasmodium falciparum ORC1 homolog. Using immunofluorescence and immunoelectron microscopy, we show here that PfORC1 is expressed in the nucleus during the late trophozoite and schizont stages where maximum amount of DNA replication takes place. Homology modelling of the carboxy terminal region of PfORC1 (781-1033) using Saccharomyces pombe Cdc6/Cdc18 homolog as a template reveals the presence of a similar AAA+ type nucleotide-binding fold. This region shows ATPase activity in vitro that is important for the origin activity. To our knowledge, this is the first evidence of an individual ORC subunit that shows ATPase activity. These observations strongly suggest that PfORC1 might be involved in DNA replication initiation during the blood stage of the parasitic life cycle. PMID- 16216222 TI - Augmentation of fatality of influenza in mice by inhibition of phagocytosis. AB - Influenza virus-infected cells undergo apoptosis and become susceptible to phagocytosis by macrophages, and this leads to the inhibition of virus propagation in vitro. To assess if this were also true in vivo, mice infected with influenza A/WSN (H1N1) virus were administered with phagocytosis inhibitors and examined for the progress of influenza. Administration of the inhibitors caused a decrease in the level of phagocytosis observed with bronchoalveolar lavage cells. We found that both the lethality in mice and the extent of inflammation in the lung were augmented in those mice. These results suggest that phagocytosis of virus-infected cells helps suppress the progress of influenza in mice. PMID- 16216223 TI - The metal-responsive transcription factor-1 contributes to HIF-1 activation during hypoxic stress. AB - Hypoxia-inducible factor-1 (HIF-1), the major transcriptional regulator of the mammalian cellular response to low oxygen (hypoxia), is embedded within a complex network of signaling pathways. We have been investigating the importance of another stress-responsive transcription factor, MTF-1, for the adaptation of cells to hypoxia. This article reports that MTF-1 plays a central role in hypoxic cells by contributing to HIF-1 activity. Loss of MTF-1 in transformed Mtf1 null mouse embryonic fibroblasts (MEFs) results in an attenuation of nuclear HIF 1alpha protein accumulation, HIF-1 transcriptional activity, and expression of an established HIF-1 target gene, glucose transporter-1 (Glut1). Mtf1 null (Mtf1 KO) MEFs also have constitutively higher levels of both glutathione (GSH) and the rate-limiting enzyme involved in GSH synthesis--glutamate cysteine ligase catalytic subunit--than wild type cells. The altered cellular redox state arising from increased GSH may perturb oxygen-sensing mechanisms in hypoxic Mtf1 KO cells and decrease the accumulation of HIF-1alpha protein. Together, these novel findings define a role for MTF-1 in the regulation of HIF-1 activity. PMID- 16216224 TI - Plasma membrane calcium-ATPase 2 and 4 in human breast cancer cell lines. AB - There is evidence to suggest that plasma membrane Ca2+-ATPase (PMCA) isoforms are important mediators of mammary gland physiology. PMCA2 in particular is upregulated extensively during lactation. Expression of other isoforms such as PMCA4 may influence mammary gland epithelial cell proliferation and aberrant regulation of PMCA isoform expression may lead or contribute to mammary gland pathophysiology in the form of breast cancers. To explore whether PMCA2 and PMCA4 expression may be deregulated in breast cancer, we compared mRNA expression of these PMCA isoforms in tumorigenic and non-tumorigenic human breast epithelial cell lines using real time RT-PCR. PMCA2 mRNA has a higher level of expression in some breast cancer cell lines and is overexpressed more than 100-fold in ZR-75-1 cells, compared to non-tumorigenic 184B5 cells. Although differences in PMCA4 mRNA levels were observed between breast cell lines, they were not of the magnitude observed for PMCA2. We conclude that PMCA2 mRNA can be highly overexpressed in some breast cancer cells. The significance of PMCA2 overexpression on tumorigenicity and its possible correlation with other properties such as invasiveness requires further study. PMID- 16216225 TI - Mitochondria-targeted peptide prevents mitochondrial depolarization and apoptosis induced by tert-butyl hydroperoxide in neuronal cell lines. AB - Oxidative stress and mitochondrial oxidative damage have been implicated in aging and many common diseases. Mitochondria are a primary source of reactive oxygen species (ROS) in the cell, and are particularly susceptible to oxidative damage. Oxidative damage to mitochondria results in mitochondrial permeability transition (MPT), mitochondrial depolarization, further ROS production, swelling, and release of cytochrome c (cyt c). Cytosolic cyt c triggers apoptosis by activating the caspase cascade. In the present work, we examined the ability of a novel cell penetrating, mitochondria-targeted peptide antioxidant in protecting against oxidant-induced mitochondrial dysfunction and apoptosis in two neuronal cell lines. Treatment with tert-butyl hydroperoxide (tBHP) for 24 h resulted in lipid peroxidation and significant cell death via apoptosis in both N2A and SH-SY5Y cells, with phosphatidylserine translocation, nuclear condensation and increased caspase activity. Cells treated with tBHP showed significant increase in intracellular ROS, mitochondrial depolarization and reduced mitochondrial viability. Concurrent treatment with <1 nM SS-31 (D-Arg-Dmt-Lys-Phe-NH2; Dmt = 2',6'-dimethyltyrosine) significantly decreased intracellular ROS, increased mitochondrial potential, and prevented tBHP-induced apoptosis. The remarkable potency of SS-31 can be explained by its extensive cellular uptake and selective partitioning into mitochondria. Intracellular concentrations of [3H]SS-31 were 6 fold higher than extracellular concentrations. Studies using isolated mitochondria revealed that [3H]SS-31 was concentrated approximately 5000-fold in the mitochondrial pellet. By concentrating in the inner mitochondrial membrane, SS-31 is localized to the site of ROS production, and can therefore protect against mitochondrial oxidative damage and further ROS production. SS-31 represents a novel platform of mitochondria-targeted antioxidants with broad therapeutic potential. PMID- 16216226 TI - Inhibitory modulation of ATP-sensitive potassium channels by gallate-ester moiety of (-)-epigallocatechin-3-gallate. AB - (-)-Epigallocatechin-3-gallate (EGCG), a major polyphenolic substance found in green tea, is well recognized to be beneficial for human health. However, it is still controversial as to what dose of this compound is indeed good for human health. Though some recent studies have interestingly reported various beneficial effects of EGCG in cell culture system, however, plasma levels of EGCG attainable by oral regular intake in humans are normally in nanomolar range. However, potential side effects of EGCG when administered parenterally at higher concentration have not been thoroughly tested. Here, we evaluated the effect of EGCG on ATP-sensitive potassium (K(ATP)) channels expressed in Xenopus oocytes. EGCG inhibited the activity of the Kir6.2/SUR1 and Kir6.2DeltaC36 channels with IC(50) of 142+/-37 and 19.9+/-1.7microM, respectively. Inhibition of EGCG was also observed in Kir6.2/SUR2A or Kir6.2/SUR2B channels. Notably, (-)-epicatechin 3-gallate (ECG), another major polyphenolic substance in green tea, was found to reduce the channel activity with greater potency than EGCG. In contrast to EGCG and ECG, which have the gallic acid-ester moiety in their own structures, (-) epigallocatechin and (-)-epicatechin exhibited very weak inhibition of the K(ATP) channel. Collectively, these results suggest that the gallate-ester moiety of epicatechins may be critical for inhibiting the K(ATP) channel activity via the pore-forming subunit Kir6.2 and this may be a possible mechanism by which green tea extracts or EGCG may cause unexpected side effects at micromolar plasma level. PMID- 16216228 TI - Action pattern of Valencia orange PME de-esterification of high methoxyl pectin and characterization of modified pectins. AB - Valencia pectinmethylesterase (PME) fractions, B-PME, containing 36 and 13 kDa protein bands and U-PME, containing a 36 and 27 kDa protein bands, were used to de-esterify original pectin (O-Pec) from 73% degree of esterification (%DE) to 63% (B-Pec) and 61% DE (U-Pec), respectively. Most O-Pec eluted from ion exchange chromatography at low salt concentration and a smaller component eluted at higher ionic strength. B-Pec and U-Pec eluted as one broad peak at higher ionic strength. PME modification did not change molecular weight: O-pectin (134,000 g/mol), U-Pec (133,850 g/mol), and B-Pec (132,250 g/mol). The NMR signal of GG and GGG increased after modification, whereas the signal of EE and EEE decreased. The negative zeta-potential increased with pH for all pectins. U-PME and B-PME created differently modified pectins that vary in degree and length of multiple attacks and fraction of the pectin population that was modified. PMID- 16216227 TI - Posterior hypothalamus cholinergic stimulation-induced activation of anterior hypothalamic area neurons is enhanced in spontaneously hypertensive rats. AB - We have previously demonstrated that some neurons in the anterior hypothalamic area (AHA) are tonically activated by endogenous angiotensins in rats and that activities of these AHA neurons are enhanced in spontaneously hypertensive rats (SHR). In addition, we have demonstrated that cholinergic mechanisms in the posterior hypothalamic nucleus (PHN) are involved in the activation of AHA angiotensin-II-sensitive neurons. It has been suggested that cholinergic function in the posterior hypothalamus is enhanced in SHR and that this hyperactivity plays a role in hypertension in SHR. In the present study, we examined whether the PHN cholinergic stimulation-induced activation of AHA angiotensin-II sensitive neurons is altered in SHR. Male 15- to 16-week-old SHR and age-matched Wistar Kyoto rats (WKY) were anesthetized and artificially ventilated. Extracellular potentials were recorded from single neurons in the AHA. Microinjection of the cholinoceptor agonist carbachol, the cholinesterase inhibitor physostigmine and the excitatory amino acid glutamate into the PHN caused increases in firing rate of AHA angiotensin-II-sensitive neurons in anesthetized WKY and SHR. The increase in firing rate of AHA neurons induced by these drugs was enhanced in SHR as compared to WKY. The enhancement of the physostigmine-induced activation of AHA neurons in SHR was similar to that of the carbachol-induced activation of AHA neurons in SHR. The enhancement of the glutamate-induced activation of AHA neurons in SHR was similar to that of the carbachol-induced activation of AHA neurons in SHR. Microinjection of scopolamine, a cholinoceptor antagonist, into the PHN caused a small but significant decrease of firing rate of AHA angiotensin-II-sensitive neurons in SHR but not in WKY. These findings indicate that the PHN cholinergic stimulation induced activation of AHA angiotensin-II-sensitive neurons is enhanced in SHR and that PHN cholinergic mechanisms are involved in tonic activation of angiotensin II-sensitive neurons in the AHA of SHR. It appears that the enhancement of the PHN cholinergic stimulation-induced activation of AHA neurons in SHR results mainly from the enhanced neural reactivity to angiotensins in AHA neurons of SHR. PMID- 16216229 TI - Mild one-pot preparation of glycosyl bromides. AB - Mild one-pot protocols for the preparation of glycosyl bromides and alkyl bromides via in situ generation of HBr is reported here. PMID- 16216230 TI - Synthesis of the beta anomer of the spacer-equipped tetrasaccharide side chain of the major glycoprotein of the Bacillus anthracis exosporium. AB - The beta glycoside of the tetrasaccharide sequence beta-Ant-(1-->3)-alpha-l-Rhap (1-->3)-alpha-l-Rhap-(1-->2)-l-Rhap, whose aglycon allows conjugation to proteins, was synthesized for the first time. A stepwise synthetic approach was applied with thioglycosides as glycosyl donors, and the beta anomer of the compound was obtained equipped with a spacer group whose further transformation allows conjugation to suitable carriers. To synthesize the beta-anthrosyl linkage with high stereoselectivity, a linker-equipped rhamnotriose derivative was glycosylated with ethyl 4-azido-3-O-benzyl-2-O-bromoacetyl-4,6-dideoxy-1-thio beta-d-glucopyranoside. Further functionalization of the tetrasaccharide thus obtained, followed by deprotection, gave the target substance. PMID- 16216231 TI - PI 3-kinase regulates the mitochondrial transition pore in controlled reperfusion and postconditioning. AB - OBJECTIVE: We investigated whether phosphatidylinositol 3-kinase (PI3K) might regulate mitochondrial permeability transition pore (mPTP) opening in hearts reperfused with either low pressure or postconditioning. METHODS: Male Wistar rat hearts (n=72) were perfused according to the Langendorff technique, exposed to 30 min of ischemia, and assigned to one of the following groups: (1) reperfusion with normal pressure (NP; 100 cm H2O), (2) reperfusion with low pressure (LP; 70 cm H2O), or reperfusion with postconditioning, i.e. 3 episodes of 30 s reperfusion followed by 30 s of ischemia (PostC). Hearts received either the PI3K inhibitors wortmannin or LY294002, or vehicle at the onset of the 60 min reperfusion. Postischemic functional recovery was assessed by rate-pressure product (RPP), and irreversible injury by lactate dehydrogenase (LDH), creatine kinase (CK) and troponin I (TnI) release. Mitochondria were isolated from the reperfused myocardium, and Ca2+-induced mPTP opening was measured using a potentiometric method. RESULTS: Functional recovery was significantly improved in LP and PostC hearts with RPP averaging 13,880+/-810 (LP) and 17,130+/-900 mm Hgxbeats/min (PostC) versus 6450+/-500 mm Hgxbeats/min in NP hearts (p<0.01). LDH release averaged 230+/-30 and 145+/-15 IU/h/g of myocardial tissue in LP and PostC versus 340+/-10 IU/h/g in NP (p<0.05). Wortmannin and LY294002 prevented both RPP improvement and decrease in LDH, CK, and TnI release in LP and PostC groups. The Ca2+ load required to induce mPTP opening averaged 58+/-3 and 52+/-1 nmol/mg mitochondrial proteins in LP and PostC groups, respectively, versus 35+/ 4 nmol/mg in the NP group (p<0.01). Wortmannin and LY294002 prevented the beneficial effect in both the LP and PostC groups. CONCLUSION: These results suggest that PI3K regulates the opening of the mitochondrial permeability transition pore in rat hearts reperfused with low pressure or postconditioning. PMID- 16216232 TI - Functions and regulations of fibroblast growth factor signaling during embryonic development. AB - Fibroblast growth factors (FGF) are secreted molecules which function through the activation of specific tyrosine kinases receptors, the FGF receptors that transduce the signal by activating different pathways including the Ras/MAP kinase and the phospholipase-C gamma pathways. FGFs are involved in the regulation of many developmental processes including patterning, morphogenesis, differentiation, cell proliferation or migration. Such a diverse set of activities requires a tight control of the transduction signal which is achieved through the induction of different feedback inhibitors such as the Sproutys, Sef and MAP kinase phosphatase 3 which are responsible for the attenuation of FGF signals, limiting FGF activities in time and space. PMID- 16216233 TI - Epithelial membrane protein-2 regulates surface expression of alphavbeta3 integrin in the endometrium. AB - The four-transmembrane protein epithelial membrane protein-2 (EMP2) was recently identified as an endometrial protein necessary for blastocyst implantation, but the mechanism of this role is uncertain. In other cell types, EMP2 controls delivery of certain classes of proteins to the cell surface, including various integrin isoforms (a class of receptors implicated in endometrial-blastocyst interaction). Since alphavbeta3 integrin is an important endometrial molecule involved in blastocyst interaction, we evaluated the role of EMP2 in modulating integrin expression in the HEC1A endometrial cell line and endometrial epithelium in vivo. Elevation of EMP2 expression in HEC1A cells selectively increased the expression of alphavbeta3 integrin on the plasma membrane and was functional as judged by increased cell binding to an alphavbeta3 ligand, fibronectin. Conversely, reduction in EMP2 expression using an EMP2 specific ribozyme decreased the cell alphavbeta3 surface expression. The influence of EMP2 on alphavbeta3 integrin was also observed in vivo as reduction of EMP2 using ribozymes or short hairpin RNA diminished alphavbeta3 integrin expression in glandular and luminal uterine epithelium. Colocalization and coimmunoprecipitation studies suggested that EMP2 and alphavbeta3 integrin predominantly exist in a physically associated state. This study demonstrates for the first time the influence of EMP2 on alphavbeta3 surface expression and suggests that surface trafficking of integrin alphavbeta3 by EMP2 during the window of implantation may be a mechanism for its requirement in endometrial blastocyst interaction. PMID- 16216234 TI - WNT8 and BMP2B co-regulate non-axial mesoderm patterning during zebrafish gastrulation. AB - During vertebrate mesoderm formation, fates are established according to position in the dorsoventral (D/V) axis of the embryo. Initially, maternal signaling divides nascent mesoderm into axial (dorsal) and non-axial (ventral) domains. Although the subsequent subdivision of non-axial mesoderm into multiple D/V fate domains is known to involve zygotic Wnt8 and BMP signaling as well as the Vent/Vox/Ved family of transcriptional repressors, how levels of signaling activity are translated into differential regulation of fates is not well understood. To address this question, we have analyzed zebrafish embryos lacking Wnt8 and BMP2b. Zebrafish wnt8; swr (bmp2b) double mutants display a progressive loss of non-axial mesoderm and a concomitant expansion of axial mesoderm during gastrulation. Mesoderm induction and specification of the axial domain occur normally in wnt8; swr mutants, but dorsal mesoderm genes eventually come to be expressed throughout the mesoderm, suggesting that the establishment of non-axial mesoderm identity requires continual repression of dorsal mesoderm factors, including repressors of ventral genes. Loss-of-function for Vent, Vox, and Ved phenocopies the wnt8; swr mutant phenotype, consistent with Wnt8 and BMP2b maintaining non-axial mesoderm identity during gastrulation through the regulation of these three transcriptional repressors. We postulate that timely differentiation of the mesoderm requires the maintenance of non-axial mesoderm identity by Wnt8 and BMP2b at the onset of gastrulation followed by subdivision of the non-axial mesoderm into different functional domains during gastrulation. PMID- 16216235 TI - A major bristle QTL from a selected population of Drosophila uncovers the zinc finger transcription factor poils-au-dos, a repressor of achaete-scute. AB - Traditional screens aiming at identifying genes regulating development have relied on mutagenesis. Here, we describe a new gene involved in bristle development, identified through the use of natural variation and selection. Drosophila melanogaster bears a pattern of 11 macrochaetes per heminotum. From a population initially sampled in Marrakech, a strain was selected for an increased number of thoracic macrochaetes. Using recombination and single nucleotide polymorphisms, the factor responsible was mapped to a single locus on the third chromosome, poils au dos, that encodes a zinc-finger-ZAD protein. The original, as well as new, presumed null, alleles of poils au dos, is associated with ectopic achaete-scute expression that results in the additional bristles. This suggests a possible role for Poils au dos as a repressor of achaete and scute. Ectopic expression appears to be independent of the activity of known cis regulatory enhancer sequences at the achaete-scute complex that mediate activation at specific sites on the notum. The target sequences for Poils au dos activity were mapped to a 14 kb region around scute. In addition, we show that pad interacts synergistically with the repressor hairy and with Dpp signaling in posterior and anterior regions of the notum, respectively. PMID- 16216236 TI - Lim 1 is required for nephric duct extension and ureteric bud morphogenesis. AB - The nephric duct plays a central role in orchestrating the development of the mammalian urogenital system. Lim 1 is a homeobox gene required for head and urogenital development in the mouse but most Lim 1-deficient embryos die by embryonic day 10. To determine the role of Lim 1 in the development of the nephric duct, we conditionally removed Lim 1 in the nephric epithelium just after the nephric duct begins to form using a floxed allele of Lim 1 and Pax2-cre transgenic mice. We report that Lim 1 conditional knockout mice have renal hypoplasia and hydronephrosis. Developmental studies revealed that the caudal portion of the nephric duct did not reach the urogenital sinus at embryonic day 10.5, formation of the ureteric bud was delayed, the ureteric bud was smaller and branching of the ureteric bud reduced. We also found that the nephric duct was generally not maintained and extension of the Mullerian duct inhibited. Molecular analysis indicated that Pax2 was expressed normally but the expression of Wnt9b and E-cadherin in the nephric duct was markedly altered. These results suggest that Lim 1 influences nephric duct extension and ureteric bud outgrowth by regulating and or maintaining the differentiation of the nephric epithelium. PMID- 16216237 TI - Xenopus Xpat protein is a major component of germ plasm and may function in its organisation and positioning. AB - In many animals, including Drosophila, C. elegans, zebrafish and Xenopus, the germ line is specified by maternal determinants localised in a distinct cytoplasmic structure called the germ plasm. This is consists of dense granules, mitochondria, and specific localised RNAs. We have characterised the expression and properties of the protein encoded by Xpat, an RNA localised to the germ plasm of Xenopus. Immunofluorescence and immunoblotting showed that this novel protein is itself a major constituent of germ plasm throughout oogenesis and early development, although it is also present in other regions of oocytes and embryos, including their nuclei. We found that an Xpat-GFP fusion protein can localise correctly in cultured oocytes, in early oocytes to the 'mitochondrial cloud', from which germ plasm originates, and in later oocytes to the vegetal cortex. The localisation process was microtubule-dependent, while cortical anchoring required microfilaments. Xpat-GFP expressed in late stage oocytes assembled into circular fields of multi-particulate structures resembling endogenous fields of germ plasm islands. Furthermore these structures could be induced to form at ectopic sites by manipulation of culture conditions. Ectopic Xpat-GFP islands were able to recruit mitochondria, a major germ plasm component. These data suggest that Xpat protein has an important role in Xenopus germ plasm formation, positioning and maintenance. PMID- 16216238 TI - Pharmacologic approaches to butterfly wing patterning: sulfated polysaccharides mimic or antagonize cold shock and alter the interpretation of gradients of positional information. AB - Butterflies produce complex and diverse wing patterns by mechanisms that are generally unknown. We have employed a pharmacological approach to explore the molecular mechanisms of pattern formation. In a screen of over 200 compounds injected into developing Junonia coenia pupae, we identified several specific sulfated polysaccharides that caused widespread, dose-dependent effects on adult wing patterns. These compounds were well tolerated and permitted butterflies to eclose normally and take flight at moderate levels of effect. Heparin and closely related chondroitin sulfates caused stage-specific expansion of distal and proximal band systems and reduction and repatterning of eyespots. Dextran sulfate and fucoidan, whose structures are widely divergent from heparin and one another, caused contraction of distal and proximal systems, but had no effect on eyespots. Nonsulfated or nonpolymeric saccharides were without effect. Pattern alterations were indistinguishable from those reported for extreme cold shock and exposure to sodium tungstate and "molsin". When administered after cold shock or coinjected with heparin, dextran sulfate reversed all patterning effects. We suggest that the primary effect of polysaccharide treatments is to alter the interpretation of gradients of positional information along the proximodistal axis of the pupal wing. PMID- 16216239 TI - Polaris and Polycystin-2 in dorsal forerunner cells and Kupffer's vesicle are required for specification of the zebrafish left-right axis. AB - Recently, it has become clear that motile cilia play a central role in initiating a left-sided signaling cascade important in establishing the LR axis during mouse and zebrafish embryogenesis. Two genes proposed to be important in this cilia mediated signaling cascade are polaris and polycystin-2 (pkd2). Polaris is involved in ciliary assembly, while Pkd2 is proposed to function as a Ca(2+) permeable cation channel. We have cloned zebrafish homologues of polaris and pkd2. Both genes are expressed in dorsal forerunner cells (DFCs) from gastrulation to early somite stages when these cells form a ciliated Kupffer's vesicle (KV). Morpholino-mediated knockdown of Polaris or Pkd2 in zebrafish results in misexpression of left-side-specific genes, including southpaw, lefty1 and lefty2, and randomization of heart and gut looping. By targeting morpholinos to DFCs/KV, we show that polaris and pkd2 are required in DFCs/KV for normal LR development. Polaris morphants have defects in KV cilia, suggesting that the laterality phenotype is due to problems in cilia function per se. We further show that expression of polaris and pkd2 is dependent on the T-box transcription factors no tail and spadetail, respectively, suggesting that these genes have a previously unrecognized role in regulating ciliary structure and function. Our data suggest that the functions of polaris and pkd2 in LR patterning are conserved between zebrafish and mice and that Kupffer's vesicle functions as a ciliated organ of asymmetry. PMID- 16216240 TI - Antinociceptive, brain-penetrating derivatives related to improgan, a non-opioid analgesic. AB - The antinociceptive profile of selected histamine H(2) and histamine H(3) receptor antagonists led to the discovery of improgan, a non-brain-penetrating analgesic agent which does not act on known histamine receptors. Because no chemical congener of improgan has yet been discovered which has both antinociceptive and brain-penetrating properties, the present study investigated the antinociceptive effects of a series of chemical compounds related to zolantidine, a brain-penetrating histamine H(2) receptor antagonist. The drugs studied presently contain the piperidinomethylphenoxy (PMPO) moiety, hypothesized to introduce brain-penetrating characteristics. Following intracerebroventricular (i.c.v.) dosing in rats, six of eight drugs produced dose- and time-related antinociception on both the tail flick and hot plate tests over a nearly eight fold range of potencies. Ataxia and other motor side effects were observed after high doses of these drugs, but two of the compounds (SKF94674 and loxtidine) produced maximal antinociception at doses which were completely devoid of these motor effects. Consistent with the hypothesis that PMPO-containing drugs are brain-penetrating analgesics, SKF94674 and another derivative (JB-9322) showed dose-dependent antinociceptive activity 15 to 30 min after systemic dosing in mice, but these effects were accompanied by seizures and death beginning 45 min after dosing. Other drugs showed a similar pattern of antinociceptive and toxic effects. In addition, loxtidine produced seizures without antinociception, whereas zolantidine produced neither effect after systemic dosing in mice. Although several of the drugs tested have histamine H(2) receptor antagonist activity, neither the antinociception nor the toxicity was correlated with histamine H(2) receptor activity. The present results are the first to demonstrate the existence of brain-penetrating antinociceptive agents chemically related to zolantidine and improgan, but further studies are needed to understand the mechanisms of both the pain relief and toxicity produced by these agents. PMID- 16216241 TI - Intracellular formation of collagen microfibrils in granulation tissue. AB - It is important to determine the biosynthesis process of collagen fibers to elucidate the mechanism by which granulation tissue is induced after injury. The purpose of this study is to investigate whether collagen microfibrils can be formed not only outside but also inside a cell. Fibroblast-like cells in granulation tissue resulting from incision and ligation were examined. The cells possessed vesicles containing collagen microfibrils. The vesicles were present in connection with Golgi apparatus or the rough endoplasmic reticulum. Furthermore, the vesicles were exhibited to be secretory granules with the secretory granule marker Rab3A. The fibroblast-like cells were also indicated to be myofibroblasts, using conventional transmission electron microscopy and immunoelectron microscopy for the myofibroblast marker alpha smooth muscle actin. In conclusion, it was demonstrated that collagen microfibrils could be formed in the cell in the case of collagen fiber overproduction. PMID- 16216242 TI - Overexpression of MyoD-inducible lysosomal sialidase (neu1) inhibits myogenesis in C2C12 cells. AB - Lysosomal sialidase, encoded by neu1, is required for the removal of terminal sialic acid residues from a variety of sialoglycoconjugates. In humans, deficiency of this enzyme results in the inborn error of metabolism sialidosis, characterized by the accumulation of sialoglycoconjugates within the nervous system and in peripheral organs. A subset of sialidosis patients present with symptoms of profound muscle dysfunction, including progressive muscular atrophy. We have previously shown that the 5' regulatory region of murine neu1 is typical of skeletal muscle-specific genes due to the presence of several E-boxes and its responsiveness to stimulation by muscle regulatory factors (MRFs) such as MyoD. Here, we report that sialidase activity is increased 6-fold during the first 24 h of differentiation of C2C12 myoblasts followed by an attenuation to pre differentiation levels by 48 h. We demonstrate that the lysosomal sialidase promoter is highly upregulated by MyoD through a mechanism that is dependent on the MyoD chromatin remodeling domain. We also show that the sialidase promoter is repressed by activated MEK. Inappropriate overexpression of sialidase 48 h after the onset of differentiation results in downregulation of myogenin as well as myosin heavy chain expression and in a halt of the differentiation cascade. This study indicates that lysosomal sialidase is a potent regulator of the early stages of myogenesis. PMID- 16216243 TI - TNF-alpha promotes cell survival through stimulation of K+ channel and NFkappaB activity in corneal epithelial cells. AB - Tumor necrosis factor (TNF-alpha) in various cell types induces either cell death or mitogenesis through different signaling pathways. In the present study, we determined in human corneal epithelial cells how TNF-alpha also promotes cell survival. Human corneal epithelial (HCE) cells were cultured in DMEM/F-12 medium containing 10% FBS. TNF-alpha stimulation induced activation of a voltage-gated K+ channel detected by measuring single channel activity using patch clamp techniques. The effect of TNF-alpha on downstream events included NFkappaB nuclear translocation and increases in DNA binding activities, but did not elicit ERK, JNK, or p38 limb signaling activation. TNF-alpha induced increases in p21 expression resulting in partial cell cycle attenuation in the G1 phase. Cell cycle progression was also mapped by flow cytometer analysis. Blockade of TNF alpha-induced K+ channel activity effectively prevented NFkappaB nuclear translocation and binding to DNA, diminishing the cell-survival protective effect of TNF-alpha. In conclusion, TNF-alpha promotes survival of HCE cells through sequential stimulation of K+ channel and NFkappaB activities. This response to TNF-alpha is dependent on stimulating K+ channel activity because following suppression of K+ channel activity TNF-alpha failed to activate NFkappaB nuclear translocation and binding to nuclear DNA. PMID- 16216244 TI - Double-stranded RNA-dependent protein kinase is required for bone calcification in MC3T3-E1 cells in vitro. AB - In this study, we demonstrated that double-stranded RNA-dependent protein kinase (PKR) is required for the calcification of osteoblasts via the signal transducers and activators of transcription 1alpha (STAT1alpha) signaling in vitro. A dominant-negative mutant PKR cDNA, in which the amino acid lysine at 296 was replaced with arginine and which does not have catalytic activity, was transfected into mouse osteoblastic MC3T3-E1 cells; thereby, we established cells that stably expressed the PKR mutant gene (PKR-K/R). Phosphorylation of PKR was not stimulated by polyinosic-polycytidylic acid in the mutant cells. The PKR-K/R mutant cells exhibited up-regulated cell growth and had low alkaline phosphatase (ALP) activity. The PKR-K/R mutant cells were not able to form bone nodules in vitro. In the PKR-K/R mutant cells, runt-related gene 2 (Runx2)-mediated transcription decreased compared with the levels in the control cells. The expression of STAT1alpha protein increased and the protein was translocated to the nucleus in the PKR-K/R mutant cells. When the expression of STAT1alpha protein in PKR mutant cells was suppressed using RNAi, the activity of Runx2 mediated transcription recovered to the control level. Our results indicate that PKR is a stimulator of Runx2 transcription and is a negative modulator of STAT1alpha expression. Our findings also suggest that PKR plays important roles in the differentiation and calcification of osteoblasts by modulating STAT1alpha and/or Runx2 expression. PMID- 16216245 TI - Progesterone inhibits human breast cancer cell growth through transcriptional upregulation of the cyclin-dependent kinase inhibitor p27Kip1 gene. AB - The effects of progesterone derivatives on breast cancer development are still controversial, probably accounting for their biphasic, opposed effects on mammary cell-cycle regulation. Here, we demonstrate in vitro that the growth-inhibitory effects of progesterone on breast cancer T-47D cells require the transcriptional upregulation of the cyclin-dependent kinase inhibitor p27(Kip1) (p27) gene. A statistical analysis of human tumor biopsies further indicates that p27 mRNA levels correlate to progesterone receptor (PR) levels. Moreover, p27 gene expression is inversely associated with tumor aggressiveness, and is a prognostic factor of favorable disease outcome. Thus, progesterone derivatives selectively activating the p27 gene promoter could be promising drugs against breast cancer progression. PMID- 16216246 TI - Nucleotide sequence and embryonic expression of quail and duck Sox9 genes. AB - Sox9 is a member of the Sry-type HMG-box (Sox) gene family. It encodes a transcription factor and is thought to be important for sexual differentiation in chicken. In the present study we have isolated Sox9 cDNAs from quail and duck, and examined the expression patterns of the corresponding genes in early embryonic gonads by whole-mount in situ hybridization. We developed a polymerase chain reaction-based protocol to identify the sex of quail and duck embryos before its morphological manifestation. Sox9 expression was first detected on days 5 and 7 in the gonads of male quail and duck embryos, respectively, and was not apparent in female gonads at these stages. These expression patterns are similar to that of chicken Sox9. Our results thus suggest that the expression of quail and duck Sox9 is associated with testis differentiation. PMID- 16216247 TI - Cloning and characterisation of a Heterodera glycines aminopeptidase cDNA. AB - An aminopeptidase full-length cDNA (Hg-amp-1) was cloned from the adult female soybean cyst nematode Heterodera glycines by heterologous screening of a cDNA library with a Caenorhabditis elegans EST sequence. The predicted open reading frame encoded an 882-amino acid protein containing the conserved zinc-binding domain and GAMEN motif that are characteristic of M1 family aminopeptidases. The putative protein lacks any subcellular targeting signals and displays strong similarity to puromycin-sensitive aminopeptidases from C. elegans, Drosophila and mammals. Hg-amp-1 is expressed in juvenile nematodes and both male and female adults, with highest expression in gravid females. In situ mRNA hybridisation localised the Hg-amp-1 transcript to the genital primordium of pre-parasitic juvenile nematodes and the reproductive tract of adult females. Suppression of Hg amp-1 transcript level by RNA-interference led to a 61% reduction in the number of female nematodes parasitising soybean roots 21 days post infection with infective juvenile nematodes that had been exposed to double-stranded RNA. PMID- 16216249 TI - Compound heterozygosity (G71R/R140H) in the lecithin:cholesterol acyltransferase (LCAT) gene results in an intermediate phenotype between LCAT-deficiency and fish eye disease. AB - The esterification of free cholesterol (FC) in plasma, catalyzed by the enzyme lecithin:cholesterol acyltransferase (LCAT; EC 2.3.1.43), is a key process in lipoprotein metabolism. The resulting cholesteryl esters (CE) represent the main core lipids of low (LDL) and high density lipoproteins (HDL). Primary (familial) LCAT-deficiency (FLD) is a rare autosomal recessive genetic disease caused by the complete or near absence of LCAT activity. In fish-eye disease (FED), residual LCAT activity is still detectable. Here, we describe a 32-year-old patient with corneal opacity, very low LCAT activity, reduced amounts of CE (low HDL cholesterol level), and elevated triglyceride (TG) values. The lipoprotein pattern was abnormal with regard to lipoprotein composition and concentration, but distinct lipoprotein classes were still present. Despite of typical features of glomerular proteinuria, creatinine clearance was normal. DNA sequencing and restiction fragment analyses revealed two separate mutations in the patient's LCAT gene: a previously described G to A transition in exon 4 converting Arg140 to His, inherited from his mother, and a novel G to C transversion in exon 2 converting Gly71 to Arg, inherited from his father, indicating that M.P. was a compound heterozygote. Determination of enzyme activities of recombinant LCAT proteins obtained upon transfection of COS-7 cells with plasmids containing G71R LCAT or wild-type LCAT cDNA revealed very low alpha- and absence of beta-LCAT activity for the G71R mutant. The identification of the novel G71R LCAT mutation supports the proposed molecular model for the enzyme implying that the "lid" domain at residues 50-74 is involved in enzyme:substrate interaction. Our data are in line with the hypothesis that a key event in the etiology of FLD is the loss of distinct lipoprotein fractions. PMID- 16216248 TI - Cyclic nucleotide kinases and tachyzoite-bradyzoite transition in Toxoplasma gondii. AB - The ability of Toxoplasma gondii to cycle between the tachyzoite and bradyzoite life stages in intermediate hosts is key to parasite survival and the pathogenesis of toxoplasmosis. Studies from a number of laboratories indicate that differentiation in T. gondii is a stress-induced phenomenon. The signalling pathways or molecular mechanisms that control formation of the latent bradyzoite stage are unknown and specific effectors of differentiation have not been identified. We engineered a reporter parasite to facilitate simultaneous comparison of differentiation and replication after various treatments. Chloramphenicol acetyltransferase (CAT), expressed constitutively from the alpha tubulin promoter (TUB1), was used to quantitate parasite number. beta galactosidase (beta-GAL), expressed from a bradyzoite specific promoter (BAG1), was used as a measure of bradyzoite gene expression. Sodium nitroprusside, a well known inducer of bradyzoite differentiation, reduced reporter parasite replication and caused bradyzoite differentiation. Stress-induced differentiation in many other pathogens is regulated by cyclic nucleotide kinases. Specific inhibitors of the cAMP dependent protein kinase and apicomplexan cGMP dependent protein kinase inhibited replication and induced differentiation. The beta GAL/CAT reporter parasite provides a method to quantify and compare agents that cause differentiation in T. gondii. PMID- 16216250 TI - Cholestanol: a serum marker to guide LDL cholesterol-lowering therapy. AB - Statins have been the mainstay of lipid-lowering therapy since their introduction. However, as lower LDL cholesterol targets are sought, adjunct therapies are becoming increasingly important. Few patients reach new targets with statin monotherapy. We propose that the cholestanol:cholesterol ratio can be used to guide lipid-lowering therapy and result in greater numbers of patients reaching target LDL cholesterol. By determining whether a patient is mainly a synthesizer or absorber of cholesterol, customized regimens can be used and are expected to improve patient outcomes and minimize costs of treatment. PMID- 16216251 TI - Muscles that support the body also modulate forward progression during walking. AB - The purpose of this study was to characterize the contributions of individual muscles to forward progression and vertical support during walking. We systematically perturbed the forces in 54 muscles during a three-dimensional simulation of walking, and computed the changes in fore-aft and vertical accelerations of the body mass center due to the altered muscle forces during the stance phase. Our results indicate that muscles that provided most of the vertical acceleration (i.e., support) also decreased the forward speed of the mass center during the first half of stance (vasti and gluteus maximus). Similarly, muscles that supported the body also propelled it forward during the second half of stance (soleus and gastrocnemius). The gluteus medius was important for generating both forward progression and support, especially during single-limb stance. These findings suggest that a relatively small group of muscles provides most of the forward progression and support needed for normal walking. The results also suggest that walking dynamics are influenced by non sagittal muscles, such as the gluteus medius, even though walking is primarily a sagittal-plane task. PMID- 16216252 TI - Tensile properties of single stress fibers isolated from cultured vascular smooth muscle cells. AB - Stress fibers (SFs), a contractile bundle of actin filaments, play a critical role in mechanotransduction in adherent cells; yet, the mechanical properties of SFs are poorly understood. Here, we measured tensile properties of single SFs by in vitro manipulation with cantilevers. SFs were isolated from cultured vascular smooth muscle cells with a combination of low ionic-strength extraction and detergent extraction and were stretched until breaking. The breaking force and the Young's modulus (assuming that SFs were isotropic) were, on average, 377 nN and 1.45 MPa, which were approximately 600-fold greater and three orders of magnitude lower, respectively, than those of actin filaments reported previously. Strain-induced stiffening was observed in the force-strain curve. We also found that the extracted SFs shortened to approximately 80% of the original length in an ATP-independent manner after they were dislodged from the substrate, suggesting that SFs had preexisting strain in the cytoplasm. The force required for stretching the single SFs from the zero-stress length back to the original length was approximately 10 nN, which was comparable with the traction force level applied by adherent cells at single adhesion sites to maintain cell integrity. These results suggest that SFs can bear intracellular stresses that may affect overall cell mechanical properties and will impact interpretation of intracellular stress distribution and force-transmission mechanism in adherent cells. PMID- 16216253 TI - Determination of Vitamin B12 in food products and in premixes by reversed-phase high performance liquid chromatography and immunoaffinity extraction. AB - A new, faster and simple method to quantify Vitamin B12, both in foods and in premixes, by reversed-phase liquid chromatography with UV detection has been developed. Vitamin B12 was extracted from food products with 50 mM sodium acetate buffer pH 4.0 (at 100 degrees C for 35 min) in the presence of sodium cyanide, followed by a purification step on an immunoaffinity column prior to the LC analysis. An enzymatic hydrolysis (pepsin at 37 degrees C and pH 4 for 3 h) prior to the purification step efficiently released the bound Vitamin B12, and thus, allowed obtaining total Vitamin B12 content in food products. Vitamin B12 was monitored by UV at 361 nm after its separation on a reversed-phase narrow-bore column with a gradient of mobile phase made of water/acetonitrile and trifluoroacetic acid (TFA) 0.025%. The specificity of the method was demonstrated by the retention characteristics, UV spectra and by comparing the peak purity with the Vitamin B12 standard. The calibration graphs plotted with six concentrations of Vitamin B12 was linear with a regression coefficient R2 > 0.9997. The repeatability of the method was evaluated at different levels of concentration on six fortified products and the relative standard deviation (RSDr) was below 3.2%. The value of the relative standard deviation of the intermediate precision was below 5.6% (n = 4). The method was successfully applied to several food products and consistent results were obtained in comparison with microbiological assay (MBA). Our data demonstrate that the immunoaffinity columns are highly efficient for the purification of Vitamin B12 and that our HPLC could be used as an alternative method to the microbiological assay for the determination of Vitamin B12 in food products. PMID- 16216254 TI - Immobilised peptide displaying phages as affinity ligands. Purification of lactoferrin from defatted milk. AB - An affinity purification procedure for the direct purification of lactoferrin from defatted (skimmed) milk has been developed. The procedure is based on using selected phage clones expressing a peptide with high binding affinity for lactoferrin which were covalently coupled to macroporous poly(dimethylacrylamide) monolithic column. Large pore size (10-100 microm) of macroporous poly(dimethylacrylamide) makes it possible to couple long (1 microm) phage particles as ligands without any risk of blocking the monolithic column. Bound lactoferrin was eluted using 1M NaCl with a purity of >95%. The technique presents a good alternative to conventional immunoaffinity chromatography for purification of a protein of interest from complex samples due to (i) the robustness of the system in terms of recovery and ligand leakage and (ii) economical aspect in terms of low ligand cost. PMID- 16216255 TI - Profiling fatty acids in vegetable oils by reactive pyrolysis-gas chromatography with dimethyl carbonate and titanium silicate. AB - A novel methodology in on-line pyrolysis-gas chromatography (Py-GC) for the fast analysis of fatty acids in vegetable oils with minimal sample treatment and the use of non-toxic reagents is described. Pyrolysis at 500 degrees C for 10 s of sub-microgram quantity of vegetable oil dissolved in dimethyl carbonate (DMC) and in the presence of nanopowder titanium silicon oxide resulted in the production of fatty acid methyl esters (FAMEs) as unique products. Pyrolysis performed by means of a resistively heated filament pyrolyser interfaced to a GC-MS apparatus enabled the direct analysis of evolved FAMEs. The DMC/Py-GC-MS analysis was tested on soybean, coconut, linseed, walnut and olive oil and the results compared to the classical BF(3)-methanol as reference methodology. The DMC method exhibited a lower precision and was biased towards lower levels of polyunsaturated fatty acids (PUFA) in comparison to the BF(3)-methanol method, but was more advantageous in terms of reduced sample treatment, waste generation and risk factors of employed chemicals. PMID- 16216256 TI - Determination of volatile compounds in grape distillates by solid-phase extraction and gas chromatography. AB - Solid-phase extraction (SPE) procedure on octadecylsilica (C18) was developed for accumulation of volatile compounds from grape distillates. The procedure was optimised for final analysis by capillary gas chromatography. At mass concentrations in model solutions ranging from 0.1 to 50 mg/l solid-phase extraction recoveries of all analytes ranged from 69% for 2-phenylethanol to 102% for capric acid, with RSD values from 2 to 9%. SPE recoveries of internal standards to be added in the sample solution prior to extraction, higher alcohols 2-ethyl-1-hexanol and 1-undecanol, were 97 and 93%, respectively, with RSD values of 3%. Detection limits of analyzed compounds in model solutions ranged from 0.011 mg/l for isoamyl acetate to 0.037 mg/l for caproic acid. Method efficiency was tested in relation to acetic acid content, volume fraction of ethanol and possible matrix effects. A significant influence of matrix on SPE efficiency for geraniol, cis-2-hexen-1-ol and cis-3-hexen-1-ol was detected. For the same reason, 2-phenylethanol could not be determined by developed SPE method in samples of grape distillates. The developed solid-phase extraction method was successfully applied to determine the differences in volatile compound content in different grape distillates produced by the distillation of crushed, pressed and fermented grapes. PMID- 16216257 TI - Temperature-responsive stationary phase utilizing a polymer of proline derivative for hydrophobic interaction chromatography using an aqueous mobile phase. AB - A new method of chromatography is proposed, utilizing a thermo-responsive polymer carrying an amino acid ester residue for the stationary phase of high-performance liquid chromatography (HPLC). We have been investigating the new concept of chromatography, a temperature-responsive chromatography, using temperature responsive poly(N-isopropylacrylamide) (PNIPAAm)-modified surface for HPLC with a constant aqueous media as the mobile phase. In this study, we designed and synthesized thermo-responsive poly(acryloyl-L-proline methyl ester) and its copolymer with N-isopropylacrylamide (NIPAAm). Homopolymers of acryloyl-L-proline methyl ester and copolymer were prepared by the reaction of radical telomerization. These polymers underwent a reversible phase transition from water soluble forms into aggregates by changing the temperature, similar to PNIPAAm. The surface properties and functions of stationary phases modified with poly(acryloyl-L-proline methyl ester) were controlled by the external temperature. In the chromatographic system, we separated steroids and amino acids with a variety of hydrophobicities using a sole aqueous mobile phase. In contrast to a PNIPAAm-modified surface, a poly(acryloyl-L-proline methyl ester)-modified surface showed a greater affinity for hydrophobic amino acids. PMID- 16216258 TI - Artificial neural network modeling of peptide mobility and peptide mapping in capillary zone electrophoresis. AB - Recently, we have developed an artificial neural network model, which was able to predict accurately the electrophoretic mobilities of relatively small peptides. To examine the robustness of this methodology, a 3-3-1 back-propagation artificial neural network (BP-ANN) model was developed using the same inputs as the previous model, which were the Offord's charge over mass term (Q/M(2/3)), corrected steric substituent constant (E(s,c)) and molar refractivity (MR). The data set consisted of 102 peptides with a larger range of size than that of our earlier report - up to 42 amino acid residues as compared to 13 amino acids in the initial study - that also included highly charged and hydrophobic peptides. The entire data set was obtained from the published result by Janini and co workers. The results of this model are compared with those obtained using multiple linear regressions (MLR) model developed in this work and the multi variable model released by Janini et al. Better predictive ability of the BP-ANN model over the MLR indicates the non-linear characteristics of the electrophoretic mobility of peptides. The present model exhibits better robustness than the MLR models in predicting CZE mobilities of a diverse data set at different experimental conditions. To explore the utility of the ANN model in simulation of the CZE peptide maps, the profiles for the endoproteinase digests of melittin, glucagon and horse cytochrome C is studied in the present work. PMID- 16216259 TI - Insecticidal effects of Buthus occitanus tunetanus BotIT6 toxin expressed in Escherichia coli and baculovirus/insect cells. AB - BotIT6 is a neurotoxin polypeptide derived from the venom of the scorpion Buthus occitanus tunetanus (Bot). Its mature form is composed of 62 amino acids. BotIT6 has been reported to be the most potent toxin from Bot venom that has a strict selectivity for insects. Such toxin may have potential as a potent animal harmless tool against insects. Using RT-PCR, we isolated and sequenced a cDNA encoding 62 amino acid residues corresponding to the known amino acid sequence of BotIT6. We have expressed a recombinant active form of BotIT6 in significantly high amounts in Escherichia coli. We have also engineered the cDNA into the Autographa californica Nuclear Polyhedrosis Virus (AcMNPV) genome and expressed the protein under control of the polyhedrin promoter. Supernatants of AcIT6-virus infected Sf9 insect cells exhibit a typical intoxication effect when injected to Spodoptera littoralis larvae. Moreover, injection of the recombinant virus showed enhanced insecticidal potency against S. littoralis larvae compared with wild type AcMNPV. PMID- 16216260 TI - Laccase-type phenoloxidase in salivary glands and watery saliva of the green rice leafhopper, Nephotettix cincticeps. AB - The activity and composition of leafhopper saliva are important in interactions with the host rice plant, and it may play a physiological role in detoxifying toxic plant substances or ingesting sap. We have characterized diphenoloxidase in the salivary glands of Nephotettix cincticeps, its activity as a laccase, and its presence in the watery saliva with the objective of understanding its function in feeding on rice plants. Nonreducing SDS-PAGE of salivary gland homogenates with staining by the typical laccase substrate 2,2'-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) (ABTS), hydroquinone or syringaldazine revealed a band at a molecular mass of approximately 85 kDa at pH 5. A band also appeared at a molecular mass of approximately 200 kDa when the gels were treated with dopamine, L-3,4-dihydroxyphenylalanine (DOPA) or catechol at pH 7. The ABTS-oxidizing activity of the homogenates was drastically inhibited by N-hydroxyglycine, a specific inhibitor of laccase. However, the dopamine-oxidizing activity was not inhibited by N-hydroxyglycine, while it was inhibited by phenylthiourea (PTU). Thus, the salivary glands of N. cincticeps contain two types of phenoloxidases: a laccase (85 kDa) and a phenoloxidase (200 kDa). Laccase activity was detected in a holidic sucrose diet that was fed on for 16 h by two females, but only a trace of catechol oxidase activity was observed, suggesting that the laccase-type phenoloxidase was the predominant phenoloxidase secreted in watery saliva. The laccase exhibited an optimum pH of 4.75-5 in McIlvaine buffer and had a PI of 4.8. Enzyme activity was histochemically localized in V cells of the posterior lobe of the salivary glands. It remained at the same level throughout the adult stage from 2 days after eclosion. A possible function of N. cincticeps salivary laccase may be rapid oxidization of potentially toxic monolignols to nontoxic polymers during feeding on the rice plant. This is the first report proving that laccase occurs in the salivary glands of Hemiptera species and is secreted in the watery saliva. PMID- 16216261 TI - Critical transition temperature and activation energy with implications for arthropod cuticular permeability. PMID- 16216262 TI - A parasitic scuticociliate infection in the Norway lobster (Nephrops norvegicus). AB - A histophagous ciliate infection was discovered in a number of Norway lobsters (Nephrops norvegicus) from the Clyde Sea Area, Scotland. Silver-carbonate staining of cultured ciliates revealed an oral apparatus and additional structural features that are morphologically similar to scuticociliates in the genus Mesanophrys, which are known to parasitize crustaceans. However, ribosomal DNA sequences (ITS1/5.8S/ITS2) of the ciliate were identical to Orchitophyra stellarum, a parasitic scuticociliate of sea stars with a different morphology from Mesanophrys spp. and to the ciliate from N. norvegicus. Associated pathology included degeneration and necrosis of the myocardial heart muscle, and large numbers of ciliates in the gill filaments. PMID- 16216263 TI - New method for testing solar sensitivity of commercial formulations of the granulovirus of codling moth (Cydia pomonella, Tortricidae: Lepidoptera). AB - A method for screening codling moth granulovirus (CpGV) formulation sensitivity to sunlight using specially prepared half apples and a solar simulator is described. The half apple preparation allows an even coverage of virus over the surface of the fruit that would not be possible using whole apples. Leaves and artificial medium were not usable for extended periods of exposure in the solar simulator due to excess drying. Fruit was sprayed with 10(-3) and 10(-5) dilutions of three commercial formulations of CpGV (Carpovirusine, Cyd-X, and Virosoft) and infested with codling moth neonates. Half of the sprayed fruit was exposed to 650 W/m2 for 4 h in an Atlas Suntest CPS solar simulator resulting in an accumulated radiant energy of 9.36x10(6) J/m2 before they were infested with neonate codling moth larvae. Spraying non-irradiated fruit with the 10(-3) dilution of Cyd-X and Virosoft resulted in nearly 100% mortality of neonate larvae. Irradiation reduced viral activity by 71-98% at the 10(-3) dilution and by up to 32% at the 10(-5) dilution relative to non-irradiated fruit. The procedures utilized enabled good preservation of the fruit throughout the incubation period and minimized invasion of the fruit by plant pathogens and saprophytic organisms. This laboratory method for screening candidate formulations and potential UV protectants could conserve time and resources by eliminating adjuvants with less potential in laboratory tests and field testing only the most promising candidates. It also enables year-round testing. PMID- 16216264 TI - Effects of A1 adenosine receptor stimulation on the expression of genes involved in calcium homeostasis. AB - We investigated whether A(1) adenosine receptor stimulation affects expression of genes involved in calcium homeostasis, including sarcolemmal L-type Ca(2+) channel, Na(+)/Ca(2+) exchanger, sarcoplasmic reticulum (SR) Ca(2+)-ATPase, phospholamban, or ryanodine receptor. Three models of A(1) stimulation were used: i) an acute model, i.e. isolated perfused rat hearts treated for 120 min with 15 nM R-phenylisopropyladenosine (R-PIA), an A(1) receptor agonist; ii) a subacute model, i.e. rats treated with 1.5 mg/kg R-PIA e.v. and sacrificed after 24 h; iii) a transgenic model, i.e. mice overexpressing A(1) adenosine receptors. In all models gene expression was determined by RT-PCR, and oxalate-supported Ca(2+) uptake, representing SR Ca(2+) uptake, was measured in the crude homogenate. Significant increase in the expression of the phospholamban gene was observed in each model of A(1) stimulation, while the expression of the other four genes was not significantly modified. In the acute model, SR Ca(2+) uptake was unaffected, however in the subacute and transgenic models uptake rate was significantly reduced. In parallel experiments, hearts obtained from the subacute model demonstrated a significant reduction in irreversible tissue injury from 30 min of ischemia and 120 min of reperfusion. Increased resistance to ischemia has already been reported also in our transgenic model. In conclusion, A(1) adenosine receptor stimulation up-regulates phospholamban gene expression, which leads within 24 h to a reduced rate of SR Ca(2+) uptake. Changes in Ca(2+) homeostasis might contribute to the delayed cardioprotective effect of adenosine. PMID- 16216266 TI - Long-chain polyunsaturated fatty acids protect the heart against ischemia/reperfusion-induced injury via a MAPK dependent pathway. AB - The mechanisms by which long-chain dietary polyunsaturated fatty acids (PUFAs) protect against cardiovascular disease are largely unknown. The present study determines the effects of eicosapentaenoic acid (EPA) and arachidonic acid (ARA) on the response of neonatal rat cardiomyocytes to simulated ischaemia (SI) and reperfusion (R). Myocytes isolated from 1-2 day old Wistar rat hearts were cultured with or without EPA or ARA and exposed to 1 h SI followed by 30 minutes reperfusion. Apoptosis was evaluated by caspase-3 activation, poly-(ADP-ribose) polymerase (PARP) cleavage and nuclear condensation. EPA (20microM) and ARA (20microM) significantly inhibited caspase-3 activation and PARP-cleavage and reduced the apoptotic index during reperfusion. Both fatty acids significantly increased ERK phosphorylation and decreased p38 phosphorylation during reperfusion. The mechanism of action of ARA on the MAPKs was further investigated with okadaic acid (to inhibit serine-threonine phosphatases) and orthovanadate (to inhibit tyrosine phosphatases). Vanadate, but not okadaic acid, significantly reduced ARA-induced inhibition of p38 phosphorylation, suggesting the involvement a tyrosine phosphatase during SI/R. Mitogen-activated protein kinase phosphatase 1 (MKP-1), a dual-specificity phosphatase, was targeted and a significant induction of MKP-1 by ARA and EPA was observed. It was demonstrated for the first time that EPA and ARA protect neonatal cardiac myocytes from ischaemia/reperfusion-induced apoptosis through activation of ERK as well as induction of a dual-specific phosphatase, causing dephosphorylation of the pro apoptotic kinase, p38. The cardioprotective effects of EPA and ARA could also be demonstrated on the functional recovery of isolated perfused hearts subjected to global ischemia. PMID- 16216267 TI - Identification and characterization of key substructures involved in the early folding events of a (beta/alpha)8-barrel protein as studied by experimental and computational methods. AB - A number of studies have examined the structural properties of late folding intermediates of (beta/alpha)8-barrel proteins involved in tryptophan biosynthesis, whereas there is little information available about the early folding events of these proteins. To identify the contiguous polypeptide segments important to the folding of the (beta/alpha)8-barrel protein Escherichia coli N (5'-phosphoribosyl)anthranilate isomerase, we structurally characterized fragments and circularly permuted forms of the protein. We also simulated thermal unfolding of the protein using molecular dynamics. Our fragmentation experiments demonstrate that the isolated (beta/alpha)(1-4)beta5 fragment is almost as stable as the full-length protein. The far and near-UV CD spectra of this fragment are indicative of native-like secondary and tertiary structures. Structural analysis of the circularly permutated proteins shows that if the protein is cleaved within the two N-terminal betaalpha modules, the amount of secondary structure is unaffected, whereas, when cleaved within the central (beta/alpha)(3-4)beta5 segment, the protein simply cannot fold. An ensemble of the denatured structures produced by thermal unfolding simulations contains a persistent local structure comprised of beta3, beta4 and beta5. The presence of this three-stranded beta barrel suggests that it may be an important early-stage folding intermediate. Interactions found in (beta/alpha)(3-4)beta5 may be essential for the early events of ePRAI folding if they provide a nucleation site that directs folding. PMID- 16216265 TI - Contractile dysfunction in hypertrophied hearts with deficient insulin receptor signaling: possible role of reduced capillary density. AB - Diabetics have worse outcomes than nondiabetics after a variety of cardiac insults. We tested the hypothesis that impaired insulin receptor signaling in myocytes worsens cardiac remodeling and function following injury, even in the absence of hyperglycemia. Mice with cardiomyocyte-restricted knock out of the insulin receptor (CIRKO) and wild type (WT) mice were treated with isoproterenol (ISO) for 2 or 5 days. Heart rates and cardiac mass increased comparably following ISO in WT and CIRKO mice. After 5 days, WT hearts were hyperdynamic by echocardiographic and left ventricular pressure measurements. However, CIRKO hearts had a blunted increase in contractility and relaxation following ISO. Interestingly, single myocytes isolated from both CIRKO ISO and WT ISO hearts had increased cellular shortening with prolonged time to peak shortening vs. respective shams. Thus, loss of myocytes or extramyocyte factors, rather than intrinsic dysfunction of surviving myocytes, caused the blunted inotropic response in ISO treated CIRKO hearts. Indeed, CIRKO ISO mice had increased troponin release after 2 days and greater interstitial and sub-endocardial fibrosis at 5 days than did ISO WT. Apoptosis assessed by TUNEL and caspase staining was increased in CIRKO ISO compared to WT ISO hearts; however, very few of the apoptotic nuclei were clearly in cardiac myocytes. After 5 days of ISO treatment, VEGF expression was increased in WT but not in CIRKO hearts. In keeping with this finding, capillary density was reduced in CIRKO ISO relative to WT ISO. Basal expression of hypoxia-inducible factor-1alpha was lower in CIRKO vs. WT hearts and may explain the blunted VEGF response. Thus, absence of insulin receptor signaling in the cardiac myocyte worsens catecholamine-mediated myocardial injury, at least in part, via mechanisms that tend to impair myocardial blood flow and increase ischemic injury. PMID- 16216268 TI - Long-range coupling between separate docking sites in interleukin-1beta. AB - The human cytokine interleukin-1beta (IL-1beta) interacts with the interleukin type I receptor using two large docking surfaces designated A and B. Crystallographic studies reveal that a single histidine residue (His30) in IL 1beta makes critical electrostatic interactions at the receptor/ligand interface. To study the function of this residue at site A, four mutant forms of IL-1beta (H30A, H30D, H30F and H30R) were investigated. The mutation that introduces charge repulsion at His30 destabilizes the protein, but paradoxically causes the least effect on receptor binding (H30D). Mutations that enhance hydrophobic or electrostatic interactions have little effect on protein stability yet markedly impair receptor binding (H30F, H30R). All mutations can transmit effects from site A to site B, as evidenced by changes in the binding of a single-chain antibody highly specific for site B. Dihedral scalar coupling constants for the wild-type IL-1beta and the four His mutant proteins showed changes in backbone angles in residues located around site B, some approximately 30 angstroms away from His30 in site A. A comparison of native solvent exchange in wild-type and mutated IL-1beta shows transmission of local destabilization along the hydrogen bond network of the beta-sheet. Taken together, the data indicate that a single residue in site A of IL-1beta can impact stability and function through perturbations in both local and long-range contacts. PMID- 16216269 TI - Mutational analysis of the SARS virus Nsp15 endoribonuclease: identification of residues affecting hexamer formation. AB - The severe acute respiratory syndrome (SARS) coronavirus virus non-structural protein 15 is a Mn2+-dependent endoribonuclease with specificity for cleavage at uridylate residues. To better understand structural and functional characteristics of Nsp15, 22 mutant versions of Nsp15 were produced in Escherichia coli as His-tagged proteins and purified by metal-affinity and ion exchange chromatography. Nineteen of the mutants were soluble and were analyzed for enzymatic activity. Six mutants, including four at the putative active site, were significantly reduced in endoribonuclease activity. Two of the inactive mutants had unusual secondary structures compared to the wild-type protein, as measured by circular dichroism spectroscopy. Gel-filtration analysis, velocity sedimentation ultracentrifugation, and native gradient pore electrophoresis all showed that the wild-type protein exists in an equilibrium between hexamers and monomers in solution, with hexamers dominating at micromolar protein concentration, while native gradient pore electrophoresis also revealed the presence of trimers. A mutant in the N terminus of Nsp15 was impaired in hexamer formation and had low endoribonuclease activity, suggesting that oligomerization is required for endoribonuclease activity. This idea was supported by titration experiments showing that enzyme activity was strongly concentration-dependent, indicating that oligomeric Nsp15 is the active form. Three-dimensional reconstruction of negatively stained single particles of Nsp15 viewed by transmission electron microscopic analysis suggested that the six subunits were arranged as a dimer of trimers with a number of cavities or channels that may constitute RNA binding sites. PMID- 16216270 TI - Characterization and HIV-1 fusion inhibitory properties of monoclonal Fabs obtained from a human non-immune phage library selected against diverse epitopes of the ectodomain of HIV-1 gp41. AB - Using a human non-immune phage library comprising more than 10(9) functional human antibody specificities in Fab format, we have been able to select a set of eight monoclonal Fabs targeted against diverse epitopes of the ectodomain of gp41 from HIV-1. The antigens used for panning the antibodies comprised two soluble, disulfide-linked, trimeric polypeptides derived from gp41, N(CCG)-gp41 and N35(CCG)-N13. The former comprises an exposed trimeric coiled-coil of the N helices of gp41 fused in helical phase to the minimal thermostable ectodomain of gp41, while the latter comprises only the trimeric coiled-coil of N-helices. The selected Fabs were probed by Western blot analysis against four antigens: N(CCG) gp41, N35CCG-N13, N34CCG (a smaller version of N35CCG-N13), and the minimal thermostable ectodomain core of gp41 in its six-helix bundle conformation (6-HB). Three classes of Fabs were found: class A (two Fabs) interact predominantly with the 6-HB; class B (four Fabs) interact with both the 6-HB and the internal trimeric coiled-coil of N-helices; and class C (two Fabs) interact specifically with the internal trimeric coiled-coil of N-helices. The IC50 values for the Fabs, expressed as bivalent mini-antibodies, ranged from 6 microg/ml to 60 microg/ml in a quantitative vaccinia virus-based reporter gene assay for HIV-1 envelope-mediated cell fusion using the envelope from the HIV-1 T tropic strain LAV. The two most potent fusion inhibitors belonged to class B. This panel of Fabs provides a set of useful probes for studying HIV-1 envelope-mediated cell fusion and may serve as a basis for developing Fab-based anti-HIV-1 therapeutics. PMID- 16216272 TI - Crystal structure of atypical cytoplasmic ABC-ATPase SufC from Thermus thermophilus HB8. AB - SufC, a cytoplasmic ABC-ATPase, is one of the most conserved Suf proteins. SufC forms a stable complex with SufB and SufD, and the SufBCD complex interacts with other Suf proteins in the Fe-S cluster assembly. We have determined the crystal structure of SufC from Thermus thermophilus HB8 in nucleotide-free and ADP-Mg bound states at 1.7A and 1.9A resolution, respectively. The overall architecture of the SufC structure is similar to other ABC ATPases structures, but there are several specific motifs in SufC. Three residues following the end of the Walker B motif form a novel 3(10) helix which is not observed in other ABC ATPases. Due to the novel 3(10) helix, a conserved glutamate residue involved in ATP hydrolysis is flipped out. Although this unusual conformation is unfavorable for ATP hydrolysis, salt-bridges formed by conserved residues and a strong hydrogen bonding network around the novel 3(10) helix suggest that the novel 3(10) helix of SufC is a rigid conserved motif. Compared to other ABC-ATPase structures, a significant displacement occurs at a linker region between the ABC alpha/beta domain and the alpha-helical domain. The linker conformation is stabilized by a hydrophobic interaction between conserved residues around the Q loop. The molecular surfaces of SufC and the C-terminal helices of SufD (PDB code: 1VH4) suggest that the unusual linker conformation conserved among SufC proteins is probably suitable for interacting with SufB and SufD. PMID- 16216271 TI - Mutant viral polymerase in the transition of virus to error catastrophe identifies a critical site for RNA binding. AB - A foot-and-mouth disease virus (FMDV) polymerase (3D) with amino acid replacements G118D, V239M and G373D (triple DMD mutant) was obtained from a molecular clone derived from a virus population treated with ribavirin, in the transition to error catastrophe (virus extinction through lethal mutagenesis). DMD 3D was expressed in Escherichia coli, purified, and its activity compared with that of wild-type enzyme and mutant enzymes with either replacement G118D, G118A or D338A (the latter affecting the catalytic motif YGDD), generated by site directed mutagenesis. No differences among the enzymes were noted in their interaction with monoclonal antibodies specific for the FMDV polymerase. Mutant enzymes with G118D or G118A showed a 100-fold decrease in polymerization activity relative to wild-type 3D, using poly(A)/oligo(dT)15 and poly(A)/VPg as template primers, under several reaction conditions. As expected, the activity of 3D with D338A was undetectable (<0.01 times the value for wild-type 3D). DMD and the G118 mutants showed impaired binding to template-primer RNA whereas the D338A mutant showed a binding similar to wild-type 3D. Transfection of cells with FMDV RNA encoding DMD 3D resulted in selection of revertant viruses that maintained only substitutions V239M and G373D. Consistently, when infectious transcripts encoded 3D with either G118D, G118A or D338A, viruses with reversions to the wild-type sequence were isolated. The implication of G118 in template-primer binding is supported by the location of this residue in the template-binding groove of the FMDV polymerase. In addition to identifying an amino acid residue that is critical for the binding of polymerase to RNA, the results document the presence of defective genomes in the transition of virus to error catastrophe. PMID- 16216273 TI - The two DNA clamps Rad9/Rad1/Hus1 complex and proliferating cell nuclear antigen differentially regulate flap endonuclease 1 activity. AB - DNA damage leads to activation of several mechanisms such as DNA repair and cell cycle checkpoints. It is evident that these different cellular mechanisms have to be finely co-ordinated. Growing evidence suggests that the Rad9/Rad1/Hus1 cell cycle checkpoint complex (9-1-1 complex), which is recruited to DNA lesion upon DNA damage, plays a major role in DNA repair. This complex has been shown to interact with and stimulate several proteins involved in long-patch base excision repair. On the other hand, the well-characterised DNA clamp-proliferating cell nuclear antigen (PCNA) also interacts with and stimulates several of these factors. In this work, we compared the effects of the 9-1-1 complex and PCNA on flap endonuclease 1 (Fen1). Our data suggest that PCNA and the 9-1-1 complex can independently bind to and activate Fen1. Finally, acetylation of Fen1 by p300-HAT abolished the stimulatory effect of the 9-1-1 complex but not that of PCNA, suggesting a possible mechanism of regulation of this important repair pathway. PMID- 16216274 TI - Effects of donor and acceptor RNA structures on the mechanism of strand transfer by HIV-1 reverse transcriptase. AB - Template switching during reverse transcription contributes to recombination in human immunodeficiency virus type 1 (HIV-1). Our recent studies suggest that the process can occur through a multi-step mechanism involving RNase H cleavage, acceptor invasion, branch migration, and finally primer terminus transfer. In this study, we analyzed the effects of reverse transcriptase (RT)-pausing, RNase H cleavages and template structure on the transfer process. We designed a series of donor and acceptor template pairs with either minimal pause sites or with pause sites at various locations along the template. Restriction sites within the region of homology allowed efficient mapping of the location of primer terminus transfer. Blocking oligomers were used to probe the acceptor invasion site. Introduction of strong pause sites in the donor increased transfer efficiency. However, the new pauses were not necessarily associated with effective invasion. In this system, the primary invasion occurred at a region of donor cleavage associated with weak pausing. These results together with acceptor structure predictions indicated that a potential invasion site is used only in conjunction with a favorable acceptor structure. Stabilizing acceptor structure at the predicted invasion region lowered the transfer efficiency, supporting this conclusion. Differing from previous studies, terminus transfer occurred at a short distance from the invasion site. Introduction of structure into the acceptor template shifted the location of terminus transfer. Nucleocapsid protein, which can improve cDNA-acceptor interactions, increased transfer efficiency with some shift of terminus transfer closer to the invasion site. Overall results support that the acceptor structure has a major influence on the efficiency and position of the invasion and terminus transfer steps. PMID- 16216275 TI - On the joint valuation of averting fatal and severe injuries in highway accidents. AB - INTRODUCTION: To evaluate the benefits of road transport safety projects, planners need the monetary value of averting fatal and severe injuries. Usually, contingent valuation and risk-risk studies have been used. The contexts posed by both survey techniques do not represent the choice situation a driver faces when having to choose among alternative routes with different levels of safety. METHOD: We set up a stated choice web page survey in which individuals had to choose between two routes for a hypothetical trip between two cities; thus implicitly revealing their preferences for safety both in terms of reducing the number of fatal victims and of severely injured victims. RESULTS: For Chilean routes we were able to estimate approximate values of US$300,000 and US$140,000 for a reduction in one fatality and one severely injured victim, respectively. IMPACTS: Our evidence could be valuable for road planners in other developing nations. PMID- 16216276 TI - Analgesic substances derived from natural products (natureceuticals). AB - From the first recorded accounts, over 7000 years ago, various forms of natural products have been utilized to treat pain disorders. Prototypical examples of such natural products are the opium poppy (Papaver soniferum) and the bark of the willow tree (Salix spp.). It was not until the 19th century when individual compounds were isolated from these substances and were determined to posses the desired effects. The known sources of these substances have been thoroughly investigated. Over the last several decades, more analgesic substances have been purified from natural products resulting in novel structural classes and mechanisms of actions. Plants and other natural products described in historical ethnobotanical and ethnopharmacological literature have become of more recent interest in drug discovery efforts. These manuscripts and reports are being utilized to aid in the identification of natural products that have been historically employed in the alleviation of pain. A large factor that has highlighted the importance of discovering novel compounds to treat pain has been in the fundamental understanding of the complex mechanisms of pain transmission in the nervous system. Nociceptive processing involves many receptor classes, enzymes and signaling pathways. The identification of novel classes of compounds from natural sources may lead to advancing the understanding of these underlying pharmacological mechanisms. With the potential of uncovering new compounds with idealistic pharmacological profiles (i.e., no side effects, no addictive potential), natural products still hold great promise for the future of drug discovery especially in the treatment of pain disorders and potentially drug addictions. PMID- 16216277 TI - Role of lupeol and lupeol linoleate on lipemic-oxidative stress in experimental hypercholesterolemia. AB - Epidemiological studies have shown that there is a positive correlation between the incidence of coronary heart disease (CHD) and the blood cholesterol level. To study the effect of plant derived triterpene, lupeol and its ester lupeol linoleate, on blood lipid status and oxidant stress in heart and hemolysate, male albino Wistar rats were fed high cholesterol diet (normal rat chow supplemented with 4% cholesterol and 1% cholic acid; HCD) for 30 days. A significant increase (p<0.05) in plasma total cholesterol (4.22 fold) and triglycerides (1.7 fold) was observed in HCD fed rats, along with elevated LDL (3.56 fold) and VLDL (1.99 fold) cholesterol and decreased HDL cholesterol (34.14%). Treatment with lupeol and its derivative normalized the lipid profile. The significant increase (p<0.05) in lipid peroxidation (LPO) was paralleled by significantly diminished (p<0.05) activities of antioxidant enzymes (SOD, CAT and GPx) and decreased (p<0.05) concentration of antioxidant molecules (GSH, Vit C and Vit E) in cardiac tissue and hemolysate of HCD fed rats. The oxidative tissue injury in hypercholesterolemic rats was substantiated by the increase in cardiac marker, serum CPK and the drop in its activity in the heart tissue. Lupeol and lupeol linoleate treatment decreased the LPO levels and increased enzymatic and nonenzymatic antioxidants. CPK activity in the treated group was comparable with that of the control. These observations highlight the beneficial effects of the triterpene, lupeol and its linoleate ester derivative, in ameliorating the lipidemic-oxidative abnormalities in the early stage of hypercholesterolemic atherosclerosis. PMID- 16216278 TI - Effects of iptakalim on extracellular glutamate and dopamine levels in the striatum of unilateral 6-hydroxydopamine-lesioned rats: a microdialysis study. AB - In a previous study, we demonstrated that iptakalim (Ipt) significantly ameliorated hypolocomotion and catalepsy induced by haloperidol and rotenone in rats. In order to further understand the mechanism(s), using a rat model of Parkinson's disease (PD) established by unilateral 6-hydroxydopamine (6-OHDA) administration to the substantia nigra pars compacta (SNpc) and reverse microdialysis techniques with high performance liquid chromatography (HPLC), we investigated the effects of Ipt on extracellular levels of glutamate, dopamine (DA) and its metabolite dihydroxyphenylacetic acid (DOPAC) in the striatum of conscious and freely moving rats. The results indicated that unilateral 6-OHDA lesioned rats have a significantly higher level of extracellular glutamate and a lower level of extracellular DOPAC in the lesioned-side of the striatum, and a lower level of extracellular DA in both sides of the striatum compared to the striatum of control rats. Ipt reduced extracellular glutamate levels in both sides of striatum of the lesioned and control rats in a concentration-dependent manner. Ipt, at lower concentrations (0.01, 0.1, 1 microM), enhanced extracellular DA levels in the lesioned-side striatum of the unilateral 6-OHDA lesioned rats, while causing no significant changes in the intact side striatum, and even a significant decline in striatum of control rats at higher concentrations of Ipt (10, 100 microM). In addition, Ipt also caused a significant decline in the extracellular DOPAC levels in the lesioned-side striatum of unilateral 6-OHDA-lesioned rats. These data suggest that the major mechanism underlying the ameliorative effects of Ipt on the behavior in 6-OHDA lesioned rats is the alteration of levels of extracellular neurotransmitters, such as glutamate and DA in the striatum of unilateral 6-OHDA-lesioned rats. PMID- 16216280 TI - Three-dimensional distributions of sewage markers in Tokyo Bay water--fluorescent whitening agents (FWAs). AB - Three-dimensional distributions of fluorescent whitening agents (FWAs: more specifically, DSBP and DAS1), which are sewage-derived water-soluble markers, were observed in Tokyo Bay water through multi-layer sampling of water at 20 locations. In summer, FWAs predominated in the surface layers, with trace but significant concentration of FWAs in bottom water due to stratification of seawater. In winter, on the other hand, FWAs were extensively mixed into the bottom layers because of the vertical mixing of seawater. In the surface layer, FWA concentrations and the DSBP/DAS1 ratio (the concentration ratio of DSBP to DAS1) were lower in summer than in winter, suggesting more efficient photodegradation of FWAs in euphotic zones during the summer due to stronger solar radiation. Horizontally, FWAs were widely distributed over the surface layer of Tokyo Bay. Surface water with DSBP concentrations above 50ng/L, corresponding to <200 times dilution of sewage effluent, was found to have spread up to 10km from the coastline. In addition, an offshore decline in FWA concentrations was observed, showing a half-distance of 10-20km. The decrease was caused by dilution by seawater of fresh water containing FWAs. The eastern part of the bay was different with respect to surface layers, with higher concentrations seen in northeastern parts. Furthermore, dispersion of combined sewer overflow (CSO)-derived water mass was observed in Tokyo Bay after heavy rain. PMID- 16216279 TI - Role of G proteins and ERK activation in hemin-induced erythroid differentiation of K562 cells. AB - Heterotrimeric G proteins which couple extracellular signals to intracellular effectors play a central role in cell growth and differentiation. The pluripotent erythroleukemic cell line K562 that acquires the capability to synthesize hemoglobin in response to a variety of agents can be used as a model system for erythroid differentiation. Using Western blot analysis and RT-PCR, we studied alterations in G protein expression accompanying hemin-induced differentiation of K562 cells. We demonstrated the presence of G(alpha s), G(alpha i2) and G(alpha q) and the absence of G(alpha i1), G(alpha o) and G(alpha 16) in K562 cells. We observed the short form of G(alpha s) to be expressed predominantly in these cells. Treatment of K562 cells with hemin resulted in an increase in the levels of G(alpha s) and G(alpha q). On the other hand, the level of G(alpha i2) was found to increase on the third day after induction with hemin, followed by a decrease to levels lower of those of uninduced cells. The mitogen-activated protein kinase ERK1/2 pathway is crucial in the control of cell proliferation and differentiation. Both Gi- and Gq-coupled receptors stimulate MAPK activation. We therefore examined the phosphorylation of ERK1/2 during hemin-induced differentiation of K562 cells. Using anti-ERK1/2 antibodies, we observed that ERK2 was primarily phosphorylated in K562 cells. ERK2 phosphorylation increased gradually until 48 h and returned to basal values by 96 h following hemin treatment. Our results suggest that changes in G protein expression and ERK2 activity are involved in hemin-induced differentiation of K562 cells. PMID- 16216281 TI - Stable isotope and trace element status of subsistence-hunted bowhead and beluga whales in Alaska and gray whales in Chukotka. AB - Tissues of bowhead, beluga, and gray whales were analyzed for Ag, Cd, Cu, Se, Zn, THg and MeHg (belugas only). Delta15N and delta13C in muscle were used to estimate trophic position and feeding habitat, respectively. Trace element concentrations in tissues were significantly different among whale species. Hepatic Ag was higher in belugas than bowheads and gray whales. Gray whales had lower Cd concentrations in liver and kidney than bowhead and belugas and a sigmoid correlation of Cd with length was noted for all whales. Renal and hepatic Se and THg were higher in belugas than in baleen whales. The hepatic molar ratio of Se:THg exceeded 1:1 in all species and was negatively correlated to body length. Hepatic and renal Zn in subsistence-harvested gray whales was lower than concentrations for stranded whales. Se:THg molar ratios and tissue concentrations of Zn may show promise as potential indicators of immune status and animal health. PMID- 16216282 TI - Are taxonomic distinctness measures compliant to other ecological indicators in assessing ecological status? AB - Assessing the ecological status, a concept implemented in the European Water Framework Directive [Directive of the European Parliament and of the Council 2000/60/EC establishing a framework for community action in the field of water policy PE-CONS 3639/1/00, 72 p.], requires the application of methods capable of distinguishing different levels of ecological quality. Somerfield and Clarke [Marine Environmental Research 43 (2003) 145-156] proposed Average Taxonomic Distinctness to be used as tool in this context. We tested the robustness of Taxonomic Distinctness measures applying it in different scenarios (estuarine eutrophication, organic pollution, and re-colonisation after physical disturbance), analysing simultaneously its compliance to other types of ecological indicators. Results show that, in most of the case studies, only Total Taxonomic Distinctness was relatively satisfactory in discriminating between disturbed situations. Other Taxonomic Distinctness measures have not proved to be more sensitive than other ecological indicators (Shannon-Wiener, Margalef, and Eco-Exergy indices). Therefore, this approach does not seem to be particularly helpful in assessing systems' ecological status with regard to the WFD implementation. PMID- 16216283 TI - Metabolites in bile of fish from Sao Sebastiao Channel, Sao Paulo, Brazil as biomarkers of exposure to petrogenic polycyclic aromatic compounds. AB - This study reports the analysis of polycyclic aromatic compound (PAC) metabolites, as biomarkers of exposure to PACs in marine environment. PAC metabolites were measured in bile samples from 14 species of demersal fish caught in the Sao Sebastiao Channel (SSC), SE Brazilian coastline. Naphthalene (NPH) equivalents, phenanthrene (PHN) equivalents, and benzo[a]pyrene (BaP) equivalents were quantified using a reverse-phase high-performance liquid chromatography coupled with fluorescence detection method. For all samples, the means (+/ standard deviation, n=37) of concentrations obtained for NPH, PHN, and BaP equivalents were, respectively, 290,000+/-200,000 ng/g, 18,000+/-14,000 ng/g, and 970+/-1900 ng/g. These results indicate recent exposure of these fish to PACs in their environment. In addition, two species (Cyclichthys spinosus and Prionotus nudigula) of fish were analyzed in order to investigate local sources of PAC contamination in the SSC and the influence of the petroleum terminal in fish caught in remote areas. The results showed that these fish species potentially migrate along the channel, especially P. nudigula. Correlations among groups of PAC metabolites indicate the same petrogenic source for NPH and PHN equivalents and a combustion source (e.g., automobile, ships) for BaP equivalents. The ratio BaP/PHN equivalents (0.05+/-0.07, n=37) confirms the predominance of petrogenic PACs for contamination by these chemicals in this region. PMID- 16216284 TI - A mathematical model for indirectly transmitted diseases. AB - We consider a mathematical model for the indirect transmission via a contaminated environment of a microparasite between two spatially distributed host populations having non-coincident spatial domains. The parasite is benign in a first population and lethal in the second one. Global existence results are given for the resulting reaction-diffusion system coupled with an ordinary differential equation. Then, invasion and persistence of the parasite are studied. A simplified model for the transmission of a hantavirus from bank vole to human populations is then analysed. PMID- 16216286 TI - Estimation and inference of R0 of an infectious pathogen by a removal method. AB - The basic reproductive ratio, R0, is a central quantity in the investigation and management of infectious pathogens. The standard model for describing stochastic epidemics is the continuous time epidemic birth-and-death process. The incidence data used to fit this model tend to be collected in discrete units (days, weeks, etc.), which makes model fitting, and estimation of R0 difficult. Discrete time epidemic models better match the time scale of data collection but make simplistic assumptions about the stochastic epidemic process. By investigating the nature of the assumptions of a discrete time epidemic model, we derive a bias corrected maximum likelihood estimate of R0 based on the chain binomial model. The resulting 'removal' estimators provide estimates of R0 and the initial susceptible population size from time series of infectious case counts. We illustrate the performance of the estimators on both simulated data and real epidemics. Lastly, we discuss methods to address data collected with observation error. PMID- 16216285 TI - A model with 'growth retardation' for the kinetic heterogeneity of tumour cell populations. AB - In the present paper we propose a continuous cell population model based on Shackney's idea of growth retardation. Cells are characterized by two state variables: the cell maturity x, 0 < or = x < or = 1, and a state variable T that identifies the rate of maturation along cell cycle. During their life span, cells can change T at random by jump transitions to T values corresponding to slower maturation rates, while at each jump the maturity x is conserved. Both the time evolution of the population and the exponential stationary solution are numerically computed. The distribution of the cell cycle transit time in asynchronous exponential growth is investigated by Monte Carlo simulation. An approximated formula for the distribution of cell cycle time is also provided. PMID- 16216287 TI - Asymptotic behaviour of solutions to abstract logistic equations. AB - We analyze the asymptotic behaviour of solutions of the abstract differential equation u'(t)=Au(t)-F(u(t))u(t)+f. Our results are applicable to models of structured population dynamics in which the state space consists of population densities with respect to the structure variables. In the equation the linear term A corresponds to internal processes independent of crowding, the nonlinear logistic term F corresponds to the influence of crowding, and the source term f corresponds to external effects. We analyze three separate cases and show that for each case the solutions stabilize in a way governed by the linear term. We illustrate the results with examples of models of structured population dynamics - a model for the proliferation of cell lines with telomere shortening, a model of proliferating and quiescent cell populations, and a model for the growth of tumour cord cell populations. PMID- 16216288 TI - A pincer-like configuration of TM2 in the human dopamine transporter is responsible for indirect effects on cocaine binding. AB - The second transmembrane segment (TM2) of DAT and other neurotransmitter transporters has been proposed to play a role in oligomerization as well as in cocaine binding. In an attempt to determine whether TM2 contributes to the binding site and/or transport pathway of DAT, we mutated to cysteine, one at a time, 25 residues in TM2 - from Phe98 to Gln122 - in an appropriate DAT background construct. Four of the mutants, F98C, G110C, P112C, and E117C, did not express at the cell surface, and G121C was inactive, despite its presence on the cell surface. Of the 21 mutants that expressed, none of the substituted cysteines reacted with MTSEA biotin-CAP, and none of the 20 functional mutants was sensitive to MTSEA or MTSET. Thus, TM2 does not appear to be water-accessible, based both on the lack of functional effects of charged MTS derivatives, and on the biochemical determination of lack of reaction with a biotinylated MTS derivative. This leads to the conclusion that TM2 does not contribute directly to the substrate-binding site or the transport pathway, and suggests that the observed effect of mutations in this region on cocaine binding is indirect. Three mutants, M106C, V107C and I108C, were crosslinked by treatment with HgCl(2). This crosslinking was inhibited by the presence of the cocaine analogue MFZ 2-12, likely due to a conformational rearrangement in TM2 upon inhibitor binding. However, the lack of crosslinking of cysteines substituted for Leu99, Leu113 and Leu120 - three of the residues that along with Met106 form a leucine heptad repeat in TM2 - makes it unlikely that this leucine repeat plays a role in symmetrical TM2 dimerization. Importantly, a high-resolution structure of LeuT, a sodium-dependent leucine transporter that is sufficiently homologous to DAT to suggest a high degree of structural similarity, became available while this manuscript was under review. We have taken advantage of this structure to explore further and interpret our experimental results in a rigorous structural context. PMID- 16216289 TI - Atypical brain laterality in adults with ADHD during dichotic listening for emotional intonation and words. AB - Few studies directly examined the nature of hemispheric specialization and interaction in ADHD. The present experiment investigated left/right brain dynamics in unmedicated right handed adults with ADHD (n = 19) and in controls (n = 19), using a dichotic listening task to assess hemispheric differences in word and emotion recognition. We also assessed how focusing attention on a single ear modulated lateralized performance and affected cross-callosal interference effects. Analysis of variance indicated that ADHD subjects showed reduced left hemisphere specialization, were better at processing emotions, and worse at processing words compared to controls. These differences were eliminated during focused attention. Finally, during presumed right hemisphere processing of linguistic stimuli, subjects with ADHD showed reduced left hemisphere interference. We concluded that ADHD subjects demonstrated greater right hemisphere and reduced left hemisphere contribution during this task relative to controls. We posit that these hemispheric differences were due to management or use of available cognitive resources rather than inherent capacity. PMID- 16216291 TI - Lack of evidence for a role for either the in utero or suckling periods in the exaggerated salt preference of the spontaneously hypertensive rat. AB - When offered as a choice with drinking water in two-bottle preference tests, the spontaneously hypertensive rats (SHR) of the Okamoto strain exhibit a marked preference for saline solutions. While this behaviour is thought to be in part genetically determined, the role of environmental influences-in particular, perinatal ones-are poorly understood. In this study, we have used combined embryo transfer and cross-fostering techniques between SHR and normotensive Wistar Kyoto (WKY) rats to delineate the relative roles of the prenatal and postnatal, suckling environment on the exaggerated saline preference of male SHR and WKY offspring at 20 weeks of age. We found, using two-bottle preference tests using water and 140 mmol/l sodium chloride solution, that neither the in utero period nor the postnatal, suckling period played a role in the development of the much larger total fluid intake (water plus saline) or saline preference (proportion of the total fluid intake taken as saline) of the SHR. We thus conclude that maternal and perinatal environmental factors do not play a major role in this behaviour and that the exaggerated saline preference of the SHR is probably largely genetically determined. PMID- 16216290 TI - In-depth analysis of spatial cognition in Williams syndrome: A critical assessment of the role of the LIMK1 gene. AB - The LIM kinase1 protein (LIMK1) is thought to be involved in neuronal development and brain function. However, its role in spatial cognition in individuals with Williams syndrome (WS) is currently ambiguous, with conflicting reports on the cognitive phenotypes of individuals who do not have classic WS but harbour partial deletions including LIMK1. Two families with partial WS deletions have been described with deficits in visuospatial cognition (Frangiskakis, J. M., Ewart, A. K., Morris, C. A., Mervis, C. B., Bertrand, & J., Robinson, et al. (1996). LIM-kinase 1 hemizygosity implicated in impaired visuospatial constructive cognition. Cell, 86, 59-69), in contrast to others with similar partial deletions who did not display spatial impairments (Tassabehji, M., Metcalfe, K., Karmiloff-Smith, A., Carette, M. J., Grant, J., & Dennis, N., et al. (1999). Williams syndrome: Use of chromosomal microdeletions as a tool to dissect cognitive and physical phenotypes. American Journal of Human Genetics, 64, 118-125). To determine the role of LIMK1 in the highly penetrant visuospatial deficits associated with classic WS, it is essential to investigate the discrepancies between the two studies. Previous research used a standardised task to measure spatial cognition, which may not pick up subtle impairments. We therefore undertook more extensive testing of the spatial cognition of two adults with partial genetic deletions in the WS critical region (LIMK1 and ELN only), who had not displayed spatial impairments in the previous study, and compared them to two high-functioning adults with WS matched on verbal ability. All participants completed a broad battery of 16 perceptual and constructive spatial tests, and the clear-cut spatial difficulties observed in the WS group were not found in the partial deletion group. These findings rule out the claim that the deletion of one copy of LIMK1 is alone sufficient to result in spatial impairment, but leave open the possibility that LIMK1 contributes to the WS cognitive deficits if deleted in combination with other genes within the WS deletion. We conclude that a deeper assessment of WS at the genetic level is required before the contribution of specific genes to phenotypic outcomes can be fully understood. PMID- 16216292 TI - mGluR2 activation of medial amygdala input impairs vomeronasal organ-mediated behavior. AB - The accessory olfactory bulb normally receives chemosensory input from the vomeronasal organ. Input from accessory bulb to medial amygdala for natural pheromone-containing conspecific chemosignals activates both anterior and posterior medial amygdala and elicits or modulates reproductive and social behavior. Here, a non-specific activation of accessory olfactory bulb by infusion of mGluR2 agonist LCCG1 in male hamsters activates immediate-early gene (Fos) expression only in anterior and not posterior medial amygdala. mGluR2 stimulation concurrently with female chemosensory stimulation produces small changes in the normal chemosensory response in medial amygdala but impairs behavior normally driven by the chemosensory input. The distribution of Fos expression, with an increase in anterior but not posterior medial amygdala, is also seen with chemosensory stimulation by chemosignals from other species, socially non relevant for hamsters, and by artificial electrical stimulation of the vomeronasal organ. We propose that the spatiotemporal pattern of amygdala input is important for eliciting normal species-specific behavior and that artificial and heterospecific stimulation fails to do so because it does not match the required pattern closely enough. Thus, modification of the pattern by addition of non-specific activation from mGluR2 agonist is sufficient to disrupt behavior normally driven by conspecific chemosensory stimulation. PMID- 16216293 TI - Community-based therapeutic care in HIV-affected populations. AB - Community-based therapeutic care (CTC) is a community-based model for delivering care to malnourished people. CTC aims to treat the majority of severely malnourished people at home, rather than in therapeutic feeding centres. This paper describes the potential of the CTC approach to provide effective care and support for people living with HIV and AIDS (PLWHA). CTC includes many of the components of a home-based care model for PLWHA. It provides outpatient treatment for common complications of HIV and AIDS, such as acute malnutrition and simple infections, and an energy-dense ready-to-use food that could be made with the appropriate balance of micronutrients for the HIV-infected patient. Through the de-centralisation of outpatient treatment sites, CTC improves accessibility by moving treatment closer to people's homes and helps to promote the sustainability of care by building on the capacity of existing health infrastructure and staff. The CTC model contains many features that are appropriate for the care and support of HIV-affected people and, in its present form, can provide effective physical care for many HIV-affected individuals. We are currently working to adapt the CTC model to make it more suitable for the support of PLWHA in the longer term. PMID- 16216294 TI - Two distinct Moloney murine leukemia virus RNAs produced from a single locus dimerize at random. AB - Two genetically distinct retroviral RNAs can be co-packaged if the RNAs are co expressed in virion producing cells. For Moloney murine leukemia virus (MLV), co packaged RNAs are not randomly selected from among all packaging-competent RNAs, but instead primarily associate as homodimers. Here, we tested the hypothesis that the distance between proviral templates might hinder RNA heterodimerization, thus generating the observed preferential homodimerization of co-expressed MLV RNAs. To do this, two genetically distinct RNAs were co-expressed from a single locus and the proportions of hetero- and homodimeric virion RNAs were determined. Unlike RNAs transcribed from two different templates, RNAs transcribed from a single locus dimerized at random. Additionally, in vitro transcription experiments suggested that MLV RNA dimerization can occur more efficiently for longer RNAs during transcription than post-synthesis. Together, these findings show that MLV RNA dimer-partner selection likely occurs either co transcriptionally or within a pool of transcripts near the proviral template. PMID- 16216295 TI - Respiratory syncytial virus nonstructural protein 2 specifically inhibits type I interferon signal transduction. AB - Human respiratory syncytial virus (RSV) inhibits type I interferon-induced gene expression by decreasing expression of signal transducer and activator of transcription (Stat)2. To identify the RSV protein that mediates effects on Stat2, airway epithelial cells were infected with vaccinia virus vectors that express single RSV proteins. Expression of RSV nonstructural (NS)2 protein alone was sufficient to decrease Stat2 levels. Furthermore, decreasing RSV NS2 levels using RNA interference in respiratory epithelial cells inhibited the RSV-mediated decrease in Stat2 expression. Airway epithelial cells were also infected with equivalent inoculums of RSV without or with single gene deletions of NS1 or NS2. RSV infection without NS2 expression did not result in decreased Stat2 levels or loss of type I interferon-dependent signaling, indicating that NS2 expression is necessary for RSV effects on Stat2. Taken together, our results indicate that NS2 regulates Stat2 levels during RSV infection, thereby modulating viral effects on interferon-dependent gene expression. PMID- 16216296 TI - Water quality parameter estimation in a distribution system under dynamic state. AB - Chlorine maintenance in distribution systems is an issue for water suppliers. The complex pipe geometry in distribution systems, the dynamic flow conditions experienced within them, and the varied nature of chlorine's reactivity make it difficult to predict chlorine levels throughout a water system. Computer-based mathematical models of water quality transport and fate within distribution systems offer a promising tool for predicting chlorine in a cost-effective manner. Nevertheless, the use of water quality models can only be effective and reliable when both hydraulics and the mechanisms of chlorine dissipation within the water system are properly defined. Bulk water decay can be measured experimentally. However, wall reaction rates are more complex to determine and must be deduced from field measurement by comparison with simulation results. The simulation-optimization model presented in this paper provides an effective tool to simplify the chlorine decay model calibration process that is often tedious. The optimization tool is based on the weighted-least-squares method solved by Gauss-Newton technique. Application of the model onto a real-life system shows that quantity, quality and location of measurement nodes play an important role in estimation of parameters. PMID- 16216297 TI - Thermal tolerance of the invasive oyster Crassostrea gigas: feasibility of heat treatment as an antifouling option. AB - Pacific oysters, Crassostrea gigas are traditionally considered shellfish of great fishery and aquaculture value. For these reasons they are introduced worldwide. Recently there has been increasing reports about the prevalence of C. gigas as biofouling organism in cooling water systems. In the absence of relevant data on the susceptibility of oysters to commonly employed antifouling techniques such as heat treatment, it was presumed that oysters would be controlled by treatment programmes directed against other major fouling organisms. The present study was carried out to test the above hypothesis, and results showed that C. gigas has an upper temperature tolerance that is much higher than other major marine fouling animals including blue mussel Mytilus edulis. Apparently, temperature regimes presently used in heat treatment of cooling water systems fouled by mussels need to be increased, if C. gigas are to be controlled effectively. Our results also indicate that previous exposure of C. gigas to sublethal high temperatures could make them more resistant to subsequent thermal treatment, an aspect that should be taken into account when heat treatment is used as a fouling control option against oyster fouling. PMID- 16216298 TI - Solvent toxicity to amphibian embryos and larvae. AB - Organic micropollutants are often damaging for aquatic organisms. Being usually hydrophobic compounds, they are often dissolved in an organic co-solvent which increases their solubility in water. The aim of this study was to study the toxicity of various solvents on embryos (protected or not by jelly coat) and on tadpoles of the common frog (Rana temporaria). Tested solvents were methanol (MeOH), methylene chloride (CH(2)Cl(2)), dimethyl sulfoxyde (DMSO), acetone (Ac) and ethanol (EtOH). Embryos exhibited higher mortality rates than tadpoles. Embryos with jelly were more sensitive to high concentration of solvents than embryos without jelly (except for acetone). According to these results, Ac, DMSO and CH(2)Cl(2) can be used as co-solvents in water to help the dissolution of micropollutants at concentration equal to or lower than 0.001 ml/l for frog embryos, and EtOH, Ac and CH(2)Cl(2) at concentration equal to or lower than 0.01 ml/l for Rana temporaria tadpoles. PMID- 16216299 TI - Polychlorinated dibenzo-p-dioxins, dibenzofurans and 'dioxin-like' PCBs in flue gas emissions from municipal waste management plants. AB - The aim of this work is to give representative data on polychlorinated dibenzo-p dioxins (PCDD), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyl (PCBs) from stack gas emissions of an urban solid waste management plant which has to comply with the limit of 0.1 ng I-TEQ/Nm3. In particular, the study is focused on 29 target compounds, the seventeen 2,3,7,8-PCDDs/Fs, four non-ortho PCBs and eight mono-ortho PCBs which configure so-called 'dioxin-like' PCBs (DL PCBs). To this end, emission measurements were performed during one year over the three operating combustion lines in a selected waste management plant. In general, accurate methodology allowed characterizing all target compounds in almost all the samples analyzed. In addition, a typical pattern for DL-PCBs is reported. The pattern presented PCB #118 to be the highest, nevertheless the figures demonstrated DL-PCBs contribution to the total TEQ around 3% being PCB #126 the most important congener due to its TEF of 0.1. Finally, remarkable differences were achieved in comparison with both environmental and biological samples such as soils, sediments, human milk or fish since these matrices may present DL-PCB contribution to the total TEQ up to 77%. PMID- 16216301 TI - Mobility of heavy metals from tailings to stream waters in a mining activity contaminated site. AB - In this paper the results of a recent characterization of Rio Piscinas (SW of Sardinia, Italy) hydrological basin are reported. In such area (about 50 km2), previous mining activities caused a serious heavy metal contamination of surface waters, groundwater, soils and biota. Acid mine drainage phenomena were observed in the area. The main sources of contamination are the tailings stored in mine tunnels and abandoned along fluvial banks. A methodological approach was adopted in order to identify relations between tailings and water contamination. Representative samples of tailings and stream sediments samples were collected. XRD analyses were performed for mineralogical characterization, while acid digestion was carried out for determining metal contents. Batch sequential leaching tests were performed in order to assess metal mobility. Also groundwater and stream water were sampled in specific locations and suitably characterized. All information collected allowed the understanding of the effect of tailings on water contamination, thus contributing to the qualitative prediction of pollution evolution on the basis of metal mobility. Finally, a potential remediation strategy of stream water is proposed. PMID- 16216300 TI - Effects of PCBs and MeSO2-PCBs on adrenocortical steroidogenesis in H295R human adrenocortical carcinoma cells. AB - Some endocrine disrupting chemicals (EDCs) in the environment have been shown to exert their biological effects through interference with steroidogenesis. In this study, the potential effects of four selected polychlorinated biphenyl (PCB) congeners (PCB101, PCB110, PCB126 and PCB149) as well as several of their environmentally-relevant methylsulfonyl-(MeSO(2)-) PCB metabolites (3'-MeSO(2) CB101, 4'-MeSO(2)-CB101, 4'-MeSO(2)-CB110, 3'-MeSO(2)-CB149 and 4'-MeSO(2)-CB149) on adrenocortical steroidogenesis were evaluated by in vitro bioassay based on the human adrenocortical carcinoma H295R cell line. The PCBs included in the study represented different structures and potential mechanisms of action. Cells were exposed for 48 h to 10 microM of each PCB congener in the presence or absence of 20% (w/w) of their corresponding MeSO(2)-PCB metabolite(s). After the chemical treatments, changes in mRNA expression of 11 steroidogenic genes (CYP11A, CYP11B1, CYP11B2, CYP17, CYP19, CYP21, 3beta-HSD1, 3beta-HSD2, 17beta HSD1, StAR and HMGR) were quantified using molecular beacon-based real-time RT PCR. Genes coding for enzymes involved in the later or final steps of steroid production (CYP11B1, CYP11B2, CYP19, 3beta-HSD1, 3beta-HSD2 and 17beta-HSD1) were up-regulated to various extents by most PCBs. The greatest transcriptional activations (2.8-29.9-fold) were elicited by PCB110 on CYP11B1, CYP11B2, 3beta HSD2 and CYP19, and PCB149 on CYP11B1, 3beta-HSD1 and 17beta-HSD1. Increased expression of these steroidogenic genes might ultimately lead to a change in hormonal balance through excessive production of steroid hormones including aldosterone, cortisol and estradiol. In addition, co-treatment with 3'- and 4' MeSO(2)-PCB149 resulted in a significant decrease in PCB149-induced 3beta-HSD1 and 17beta-HSD1 expression. This result indicates that some PCB congeners and their MeSO(2)-metabolites may affect steroidogenesis via different mechanisms. Overall, these findings suggest that PCBs and PCB metabolites can affect regulation of adrenocortical steroidogenesis. PMID- 16216302 TI - Effects of combustion and operating conditions on PCDD/PCDF emissions from power boilers burning salt-laden wood waste. AB - This paper discusses the effects of combustion conditions on PCDD/PCDF emissions from pulp and paper power boilers burning salt-laden wood waste. We found no correlation between PCDD/PCDF emissions and carbon monoxide emissions. A good correlation was, however, observed between PCDD/PCDF emissions and the concentration of stack polynuclear aromatic hydrocarbons (PAHs) in the absence of TDF addition. Thus, poor combustion conditions responsible for the formation of products of incomplete combustion (PICs), such as PAHs and PCDD/PCDF precursors, increase PCDD/PCDF emissions. PAH concentrations increased with higher boiler load and/or low oxygen concentrations at the boiler exit, probably because of lower available residence times and insufficient excess air. Our findings are consistent with the current understanding that high ash carbon content generally favours heterogeneous reactions leading to either de novo synthesis of PCDD/PCDFs or their direct formation from precursors. We also found that, in grate-fired boilers, a linear increase in the grate/lower furnace temperature produces an exponential decrease in PCDD/PCDF emissions. Although the extent of this effect appears to be mill-specific, particularly at low temperatures, the results indicate that increasing the combustion temperature may decrease PCDD/PCDF emissions. It must be noted, however, that there are other variables, such as elevated ESP and stack temperatures, a high hog salt content, the presence of large amounts of PICs and a high Cl/S ratio, which contribute to higher PCDD/PCDFs emissions. Therefore, higher combustion temperatures, by themselves, will not necessarily result in low PCDD/PCDFs emissions. PMID- 16216303 TI - The value of biodegradation screening test results for predicting the elimination of chemicals' organic carbon in waste water treatment plants. AB - Based on the test results of a large number of chemical substances, the comparability of different biodegradation test methods was assessed in terms of passing or failing threshold values commonly used for the biodegradability characterisation of chemicals. The comparison revealed a high consistency of the ready biodegradability evaluations in OECD 301 tests regardless of the analytical parameter used, i.e., by mineralisation (CO2 evolution or biological oxygen demand/BOD) or by organic carbon removal measurements. Substances meeting the ready biodegradability threshold values in OECD 301 tests exhibited a high (> 80%) carbon removal in model waste water treatment plants (OECD 303A). Results from the Modified Zahn-Wellens test (OECD 302B) representing a screening test with high sludge concentration, were also shown to be useful for the prediction of the organic carbon removal under WWTP conditions provided some prerequisites are met. The analysis of the compared data pairs formed a solid basis for the conclusion that positive results of a chemical in biodegradability screening tests can be used for a reliable prediction of their carbon-based removal in waste water treatment plants. PMID- 16216304 TI - Sources of priority substances entering an urban wastewater catchment--trace organic chemicals. AB - The implementation of new legislation such as the Water Framework Directive (WFD) requires Member States to gain a better understanding of priority substances entering surface waters. This will include inputs from wastewater treatment works as well as from other urban, industrial and agricultural sources. There is currently a lack of available data regarding the magnitude and sources of organic priority substances entering treatment works. As a consequence the concentrations of organic priority substances including PAHs, surfactants, polybrominated diphenylethers (PBDEs), diethylhexyl phthalate (DEHP), pesticides and solvents were determined in the wastewater from an urban catchment located in the UK, as part of a project undertaken for UK Water Industry Research (UKWIR). Most organic priority substances were detected in the microg/l range. Significant variations in the concentration of linear alkylbenzene sulphonate (LAS), DEHP, PBDEs and nonylphenol ethoxylates (NPEOs), in particular, were observed for the different sources. The greatest differences were evident between new and older domestic discharges. Solvent levels varied considerably reflecting use and production within the catchment. Chloroform levels were highest in domestic effluent, while trichloroethene and tetrachloroethene were detected only in commercial samples. PMID- 16216305 TI - Application of 16S rDNA-PCR amplification and DGGE fingerprinting for detection of shift in microbial community diversity in Cu-, Zn-, and Cd-contaminated paddy soils. AB - Seven soils were sampled from farmland at different distances (0.01-5 km) from a copper and zinc smelter. The total contents of heavy metals in these soils ranged from 46 to 4895 mg Cu kg-1, 96 to 1133 mg Zn kg-1, and 6.9 to 28.8 mg Cd kg-1, respectively. The available fractions were highly correlated with total contents of the metals. In order to assess the impact of combined contamination of heavy metals on soil bacterial communities, denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplicons of 16S rDNA sequence of bacteria in soil was used. Bacterial community structure was affected to some extent by heavy metals. The number of DGGE bands in soils increased with increasing distance from the copper and zinc smelter. Clustering analysis of the DGGE profiles showed that bacteria in the seven soils belonged to three clusters. Bacterial communities in three soils sampled at 0.01-0.60 km from the smelter belonged to one cluster, and those in three soils sampled at 0.8-1.2 km from the smelter belong to another cluster. Bacterial community in soil farthest from the smelter belonged to a single cluster. This study demonstrated that heavy metal contamination decreased both biomass and diversity of bacterial community in the soil. PMID- 16216306 TI - Field study of methidathion in soil amended with biosolid and a cationic surfactant under different irrigation regimes. Solute transport modeling. AB - Four experimental plots located in Granada (Spain) were used to investigate the potential movement of the insecticide methidathion during three treatments in a period of three years. To increase pesticide soil retention a municipal biosolid and the cationic surfactant, tetradecyl trimethyl ammonium bromide (TDTMA), were used as soil amendments. The presence of the insecticide was monitored in soil and water samples at different depths up to one meter. Soil solution was sampled by ceramic suction cups installed at three depths (25, 75 and 100 cm). No effect of the amendments on pesticide mobility was observed. Experimental results showed that pesticide leaching occurred in the upper soil layer. Although some sporadic high water soil concentrations were found, these were attributed to preferential flow processes. This was confirmed by the absence of high pesticide concentration in soil samples at similar depths. Pesticide mobility was mainly affected by the irrigation employed. Experimental results were compared with theoretical data simulated with the mathematical model FocusPelmo. The resemblance between theoretical and experimental soil data seems to confirm the preferential flow processes. Otherwise, the lack of fit between the soil water data were attributed to the ceramic devices employed, that could suffer an "ageing process" which would cause bias in the determinations. PMID- 16216307 TI - Kinetic assessment of persistent halogenated xenobiotics in cell culture models: comparison of mono- and poly-halogenated compounds. AB - We evaluated the suitability of single and multiple cell type cultures as model systems to characterise cellular kinetics of highly lipophilic compounds with potential ecotoxicological impact. Confluent mono-layers of human skin fibroblasts, rat astrocytoma C6 cells, non-differentiated and differentiated mouse 3T3 cells were kept in culture medium supplemented with 10% foetal calf serum. For competitive uptake experiments up to four different cell types, grown on glass sectors, were exposed for 3h to (14)C-labelled model compounds, dissolved either in organic solvents or incorporated into unilamellar lecithin liposomes. Bromo-, or chloro-benzenes, decabromodiphenylether (DBP), and dichlorodiphenyl ethylene (DDE) were tested in rather high concentration of 20 microM. Cellular toxicity was low. Compound levels were related to protein, DNA, and triglyceride contents. Cellular uptake was fast and dependent on physico chemical properties of the compounds (lipophilicity, molecular size), formulation, and cell type. Mono-halogenated benzenes showed low and similar uptake levels (=low accumulation compounds). DBP and DDE showed much higher cellular accumulations (=high accumulation compounds) except for DBP in 3T3 cells. Uptake from liposomal formulations was mostly higher than if compounds were dissolved in organic solvents. The extent of uptake correlated with the cellular content of triglycerides, except for DBP. Uptake competition between different cell types was studied in a sectorial multi-cell culture model. For low accumulation compounds negligible differences were found among C6 cells and fibroblasts. Uptake of DDE was slightly and that of DBP highly increased in fibroblasts. Well-defined cell culture systems, especially the sectorial model, are appropriate to screen for bioaccumulation and cytotoxicity of (unknown) chemical entities in vitro. PMID- 16216308 TI - Metal accumulation in eggs of the red-eared slider (Trachemys scripta elegans) in the Lower Illinois River. AB - The Illinois River is a highly utilized navigable waterway in the US Midwest, and has historically been contaminated with metal toxicants from various industrial and municipal pollution sources. Little information on metal contamination is available in the Lower Illinois River, and in particular, in the habitat of the red-eared slider (Trachemys scripta elegans) at the southern end of the river near Grafton, IL. This study was conducted to determine current levels of metal contamination in water, sediment, soil, and plants in the habitat, as well as to reveal temporal and spatial variations of metal accumulation in eggs of the red eared slider. Aluminum, Cd, Cr, Cu, Mn, Ni, Pb, V, Sn, and Zn were analyzed by inductively-coupled plasma spectroscopy. High concentrations of metals were observed in lake sediment, compared with the concentrations in water, soil, and plant tissues. Sediment Ni concentrations (mg kg(-1)) varied from 66 to 95 and Sn from 1100 to 1600. Five detectable metals in egg content were Zn (24.2 +/- 13), Al (2.2 +/- 1.2), Sn (1.8 +/- 1.1), Mn (1.1 +/- 0.6), and Cu (0.9 +/- 0.5); nine detectable metals in egg shell were Zn (6.8 +/- 3.9), Sn (3.7 +/- 3.1), Cu (1.9 +/- 1.3), Cr (1.6 +/- 1.5), V (1.6 +/- 1.4), Pb (1.3 +/- 0.7), Ni (1.3 +/- 0.9), Mn (1.0 +/- 0.8), and Cd (0.16 +/- 0.11). Zinc accumulation in egg content was significantly correlated with Zn in egg shell (r = 0.445, P < 0.002, n = 42). While significant spatial variation was observed in egg shell, metal accumulation in eggs (content and shell) collected from the same ground of turtles consecutively for 4 years did not show a significant temporal change. PMID- 16216309 TI - Distribution of alkylphenols in the Pearl River Delta and adjacent northern South China Sea, China. AB - The occurrence of alkylphenols (APs) was investigated in surface water and sediments from the Pearl River Delta and adjacent northern South China Sea. Most of the water samples contained detectable amounts of APs, ranging up to 0.628 microg l(-1) for nonylphenol (NP) and 0.068 microg l(-1) for octylphenol (OP). APs were found in all of the sediment samples with concentrations ranging from 59 to 7808 microg kg(-1) for NP and from 1 to 93 microg kg(-1) for OP. The Zhujiang River showed the highest concentrations of APs in both water and sediments. Significant decrease of APs concentrations going from the Zhujiang River to the Shiziyang River was observed. The Xijiang River contained concentrations of APs slightly higher in water but relatively lower in sediments than the Lingding Bay, which might be attributed to their different hydrodynamic and sedimentary characteristics. There was a decreasing trend of APs in water from the rivers to the estuary and further to the sea on the whole. In the Lingding Bay and its outer waters, concentrations of APs in sediments increased to a maximum and then decrease seaward, which was consistent with the distribution trend of the sediment organic carbon contents. Linear regression analyses showed the concentrations of APs were markedly correlated with the sediment organic carbon contents, indicating that the sediment organic carbon is an important factor controlling the levels of APs in sediments. PMID- 16216310 TI - Determination of mercury methylation potentials in the water column of lakes across Canada. AB - A stable isotope technique was used to trace the formation of methylmercury in lake water incubation assays at in situ conditions in five lakes across Canada. Methylation activity was only detected in the anoxic hypolimnia of lakes. The stable isotope was methylated at varying rates between lakes and depths within lakes ranging from 0.56%/day to 14.8%/day. A peak in methylation potential was typically observed just below the oxycline, which decreased with increasing depth. The depth and rates of methylation potential changed seasonally with no methylation activity occurring after fall turnover. A decrease in the sulfate concentration was concomitant with the zone of mercury methylation potential indicating the likely involvement of sulfate reducing bacteria in the methylation process. A simple correlation test between DOC concentrations and methylation rates indicated a positive relationship (r2=0.62; p=0.006; n=27). The demethylation rate constant in the anoxic hypolimnia was less than 0.12 d(-1). PMID- 16216311 TI - Mercury contaminations from historic mining to water, soil and vegetation in Lanmuchang, Guizhou, southwestern China. AB - Concentrations of total mercury and methylmercury (MeHg) were measured in soil and vegetation samples collected from a small area with a long history of Hg mining. Hg distributions were determined in stream-waters during two sampling periods. Total Hg concentrations in soil and vegetation samples were highly elevated ranging from 0.41 to 610 mg kg(-1) and from 0.02 to 55 mg kg(-1), respectively. MeHg concentrations varied from 0.41 to 8.8 microg kg(-1) in soil samples and from 0.65 to 5.5 microg kg(-1) in vegetations. The concentrations of total Hg in stream waters varied from 55.0 to 7020 ng L(-1) in the flood-flow regime and from 24.8 to 679 ng L(-1) in the base-flow regime, respectively. Average dissolved Hg concentration was 15.7 ng L(-1) in the wet season and 21.0 ng L(-1) in the dry season. However, particulate Hg was typically >70% of total Hg in the flood-flow regime. Higher concentrations of particulate Hg primarily originated from summer floods were the major pathway of Hg transportation, which were evidenced by the positive correlation between particulate Hg and total suspended solids (TSS). The contaminated soils and distribution patterns of Hg in the stream-waters may serve as an important source of Hg to the local environment in the study area. PMID- 16216312 TI - Field assessment of treatment efficacy by three methods of phosphoric acid application in lead-contaminated urban soil. AB - In situ soil treatment using phosphoric acid (H(3)PO(4)) may be an effective remedial technology for immobilizing soil Pb and reducing Pb risk to human health and ecosystem. The treatment efficacy of three H(3)PO(4) application methods was assessed in a smelter-contaminated urban soil located in the Jasper County Superfund Site, Missouri. Soil, with an average of 3529 mg Pb kg(-1) and in the 2 by 4-m plot size, was treated with H(3)PO(4) at a rate of 10 g P kg(-1) in four replicates by each of three methods: rototilling; surface application; pressure injection. Three soil cores, 2.5-cm diameter and 30-cm long, were taken from each plot before and 90 days after treatment and analyzed for soluble P, bioaccessible Pb and solid-Pb speciation. Applications of H(3)PO(4) induced the heterogeneity of soluble P in soil, with the highest concentrations in the surface. Three application methods mixed the H(3)PO(4) more effectively in the horizontals than the verticals of treated soil zone. The H(3)PO(4) applications significantly reduced Pb bioaccessibility in the soil, which was influenced by the concentrations of soil soluble P and solid-Pb species. The risk reductions of soil Pb were achieved by formation of pyromorphites or pyromorphite-like minerals. The rototilling appears to be the most effective treatment method in context of the homogeneity of soluble P and the reduction of Pb bioaccessibility in treated soil. PMID- 16216313 TI - Do mucous aggregates affect macro-zoobenthic community and mussel culture? A study in a coastal area of the Northwestern Adriatic Sea. AB - In June 2000 and July 2002, two mucous aggregation events of large proportion occurred in the Adriatic Sea. In order to assess the possible effects that the events had on the macrofauna, we studied macro-zoobenthic assemblages and mussel culture (Mytilus galloprovincialis). Structural parameters of macro-zoobenthos and growth parameters of mussels were recorded. The study area was a mussel farm located 5 nautical miles off Porto Caleri (Rovigo, Italy) in the Northern Adriatic Sea. Between May 2000 and August 2002, two sites were sampled in this area and univariate and multivariate techniques were used to describe the macrobenthic community. Two-way ANOVA showed that mean values of species number, density (individuals m(-2)) and Shannon-Wiener's diversity were significantly influenced (p<0.01) by site and period of sampling and by the interactions of these factors. The Pielou's J averages were significantly influenced only by period (p<0.01). Application of Tuckey's HSD test (p<0.05) to factors detected to be significant by ANOVA did not show significant differences between samples collected after the mucous aggregation events and the other periods. Cluster analysis and MDS ordination did not allow a clear distinction between the samples. Concerning mussels, one-way ANOVA showed that mean values of the shell length and the condition index (dry weight/shell weight) were significantly influenced (p<0.001) by the months. The mean values both for shell length and condition index were higher in 2001. The shell length trend revealed a slowing down of growth in June-July 2000 and July-August 2002, and the condition index trend showed a significant fall in samples of June 2000 and July 2002. The growth of M. galloprovincialis might be influenced negatively by mucous aggregates, whereas the soft-bottom macro-zoobenthos seems not to be directly affected by the event. PMID- 16216314 TI - Relationships between northern Adriatic Sea mucilage events and climate variability. AB - A long term analysis (1865-2002) of meteorological data collected in the Po Valley and Northern Adriatic Basin have been analysed to find possible links between variability in the climatic parameters and the phenomenon of mucilage. Seasonal anomalies of temperature, calculated as spatial mean over the Po Valley area, and anomalies of North Atlantic Oscillation were compared with the historical record of mucilage episodes. Both climatic indices were found to be positively correlated with mucilage events, suggesting a possible relationship between climatic variability and the increased appearance of mucilage aggregates. PMID- 16216315 TI - Clinical results and quality of life analysis for the MVAC combination (methotrexate, vinblastine, doxorubicin, and cisplatin) in carcinoma of the uterine cervix: A Gynecologic Oncology Group study. AB - OBJECTIVES: The Gynecologic Oncology Group (GOG) compared methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) with topotecan and cisplatin (TC) or cisplatin alone (C) in advanced cervical cancer. The primary endpoint was overall survival (OS), with response rate, progression-free survival (PFS), and quality of life (QOL) as secondary objectives. METHODS: Eligible patients were randomly allocated to receive either cisplatin 50 mg/m2 q 3 weeks (C) or cisplatin 50 mg/m2 day 1 and topotecan 0.75 mg/m2 days 1-3 q 3 weeks (TC) or methotrexate 30 mg/m2 days 1, 15, and 22, vinblastine 3 mg/m2 days 2, 15, and 22, doxorubicin 30 mg/m2 day 2, and cisplatin 70 mg/m2 day 2 q 4 weeks (MVAC). QOL was assessed at four time points using the Functional Assessment of Cancer Therapy-Cervix (FACT-Cx), Neurotoxicity Subscale (FACT/GOG-NTX subscale), and Brief Pain Inventory (BPI). RESULTS: One hundred eighty-six patients (C = 60; TC = 63; MVAC = 63) were enrolled before MVAC was closed by the GOG Data Safety Monitoring Board after four treatment-related deaths occurred on that arm. MVAC produced a 22% overall response rate (95% CI: 0.13 to 0.34) and median PFS and OS of 4.4 months and 9.4 months, respectively. Compared with C and TC, there was more hematologic toxicity with MVAC. There were no appreciable differences in QOL scores after controlling for baseline scores. CONCLUSIONS: MVAC's clinical activity tended to be similar to that of TC but with an unacceptable risk of death from sepsis at this dose and schedule. Nevertheless, QOL, as measured by these instruments, was not substantially impaired by this regimen. PMID- 16216317 TI - The AP-2gamma transcription factor is upregulated in advanced-stage ovarian carcinoma. AB - OBJECTIVE: To analyze the expression of the AP-2gamma transcription factor in ovarian borderline tumors, early-stage ovarian carcinoma and advanced-stage ovarian carcinoma, and to evaluate its prognostic role in advanced-stage tumors. METHODS: Sections from 14 normal ovaries, 75 borderline tumors, 22 FIGO stage I invasive ovarian carcinomas, and 306 advanced-stage (FIGO stages II-IV) ovarian carcinomas (42 primary tumors, 62 solid metastases, 202 effusions) were evaluated for expression of the transcription factor AP-2gamma using immunohistochemistry. Sixty-three effusions and two cell lines (SKOV-3 and OVCAR-3) were additionally studied using immunoblotting. The prognostic role of AP-2gamma in advanced-stage carcinomas was analyzed. RESULTS: AP-2gamma was detected in the nucleus of tumor cells in 28/75 (37%) borderline tumors, 13/22 (59%) FIGO stage I carcinomas, and 255/306 (83%) advanced-stage carcinomas (P < 0.001, Chi-square test). Benign ovaries were uniformly negative. Expression was largely limited to carcinoma cells in effusions. Solid lesions and effusions from advanced-stage carcinomas showed comparable expression. Immunoblotting showed AP-2gamma expression in 59/61 effusions and both cell lines. AP-2gamma expression did not correlate with survival. CONCLUSIONS: AP-2gamma expression is upregulated in advanced-stage ovarian carcinoma compared to early-stage carcinomas, borderline tumors, and the ovarian surface epithelium, and AP-2gamma is specifically localized to cancer cells in effusions, suggesting a role in tumor progression. The lack of predictive value for this transcription factor in advanced-stage disease may be related to its frequent expression. PMID- 16216319 TI - Development and evaluation of a highly sensitive human papillomavirus genotyping DNA chip. AB - OBJECTIVES: Test of human papillomavirus (HPV) is a useful adjunctive tool of Pap smear to screen cervical cancer. We have developed a novel HPV genotyping DNA chip arrayed by multiple oligonucleotide probes of both L1 and E6/E7 gene sequence of 42 types of anogenital HPV. METHODS: Consensus PCR products of L1 and E6/E7 gene sequences of HPV are hybridized to arrayed probes on the HPV chip and HPV genotypes are identified by fluorescence scanner. We have comparatively analyzed the value of HPV DNA chip and DNA sequencing in 100 cervical cancer tissues. RESULTS: Overall, 98 cervical cancer tissues were found to harbor DNA sequences of high-risk type HPVs, of which 88 (89.8%) were detected by PCR sequencing of L1 alone, 98 (100%) by PCR-sequencing of both L1 and E6/E7, and 98 (100%) by HPV DNA chip, respectively. All of the genotypes of HPV detected on sequencing analysis were also found on DNA chip analysis. HPV DNA chip was superior to direct DNA sequencing in detection of mixed infection. CONCLUSIONS: These results suggest that HPV DNA chip analysis in the present study is highly accurate for detection and genotyping of HPV and may have potential value as a robust, high-throughput screening test of uterine cervix cancer. PMID- 16216321 TI - Re: "Outcomes of stage I/II vulvar cancer patients after negative superficial inguinal lymphadenectomy". PMID- 16216320 TI - Conservative surgery for borderline ovarian tumors: a review. AB - OBJECTIVE: Borderline tumor of the ovary is an epithelial tumor with a low rate of growth and a low potential to invade or metastasize. This review will outline the most recent information regarding the molecular pathogenesis, pathology, fertility and tumor recurrence rate after conservative management of young women with early-stage borderline ovarian tumors. METHODS: We performed a MEDLINE literature search of relevant clinical trials for the scope of this review that evaluated conservative treatment of borderline ovarian tumors for young women with low-stage disease who wish to preserve their fertility. RESULTS: Recently, investigators have begun to identify subsets of patients with a worse prognosis, such as patients with aneuploid tumors. A number of oncogenes are under investigation to determine their role in the pathogenesis of borderline ovarian tumors. Previous studies have suggested the safety of conservative surgery with unilateral salpingo-oophorectomy or cystectomy for patients with stage I borderline ovarian tumors. Laparoscopic treatment of adnexal masses has proved to be a safe and effective diagnostic and therapeutic tool in the hands of experienced laparoscopists. For women who are treated conservatively, follow-up is important. Surgery remains the most effective therapy for later stage lesions. Adjuvant therapy for advanced stage of borderline ovarian tumors remains controversial. CONCLUSION: Conservative management of borderline ovarian tumors is an appropriate therapeutic option for young women with early-stage lesions who wish to preserve their childbearing potential. Available data indicate that in these patients fertility, pregnancy outcome and survival remain excellent. PMID- 16216323 TI - Stimulus gated cocaine sensitization: interoceptive drug cue control of cocaine locomotor sensitization. AB - Repeated cocaine treatments typically generate sensitization effects which are environment specific. In this study, we investigated whether drug treatments with highly selective receptor specificity can also function as contextual cues to control the expression of cocaine sensitization effects. Two experiments were conducted in which separate groups of rats (N=10) received ten paired or unpaired cocaine (10.0 mg/kg) treatments. In the experiments, autoreceptor preferring low doses of either the 5-HT1A agonist, 8-OHDPAT (8OH) (0.05 mg/kg) or the D1/D2 agonist apomorphine (APO) (0.05 mg/kg) were administered 20 min prior to cocaine administration and test environment placement (paired treatment). Under these conditions, the drug cues generated by the 8OH/APO treatments were associated with the cocaine stimulant effect in the test environment. The unpaired treatment groups received the same drug treatments but the cocaine was administered after testing, in the homecage. Consequently, for these groups, the 8OH/APO drug cues generated by the drug treatments would not become associated with the cocaine stimulant effect in the test environment. Critically, both 8OH and APO pretreatments elicited equivalent unconditioned response effects which were opposite to the cocaine unconditioned response effects; that is, behavioral inhibition vs. behavioral stimulation. Initially, the 8OH and APO pretreatments prevented the locomotor stimulant effects of cocaine; but, these inhibitory effects were reversed in the paired groups with repeated cocaine treatments, consistent with the emergence of cocaine sensitization effects. In the unpaired 8OH and APO pretreatment groups, behavioral suppression persisted throughout the treatment protocol. Subsequently, paired and unpaired groups were compared in four conditioning/sensitization tests. The conditioning tests included: a saline/saline test; and a 8OH/saline test (Experiment 1); and, a saline/saline test and a APO/saline test (Experiment 2). There were no paired/unpaired group differences in these conditioning tests. The sensitization tests included: a saline/cocaine test; and a 8OH/cocaine test (Experiment 1); and, a saline/cocaine test and a APO/cocaine test (Experiment 2). There were no paired/unpaired group differences in the saline/cocaine test for sensitization but paired/unpaired group differences were found in both the 8OH/cocaine and APO/cocaine sensitization tests. In these tests the paired but not the unpaired groups exhibited cocaine locomotor sensitization effects. Critically when, in an additional test, the pretreatments in the cocaine tests were reversed (i.e., 8OH paired group received APO and APO paired group received 8OH prior to cocaine), then there was no evidence for cocaine sensitization. Since the 8OH/APO pretreatments had equivalent inhibitory response effects, it was the stimulus properties of these drugs which controlled the expression of the cocaine locomotor sensitization effects. These findings support the critical role of associative processes in the stimulus-gating of psychostimulant drug sensitization. Importantly, this report incorporates a new methodology in which context can be specified in terms of highly specific brain receptor targets rather than in terms of global environmental situational cues. PMID- 16216322 TI - KKHA-761, a potent D3 receptor antagonist with high 5-HT1A receptor affinity, exhibits antipsychotic properties in animal models of schizophrenia. AB - KKHA-761, 1-{4-[3-(3,4-dimethoxy-phenyl)-isoxazol-5-yl]-butyl}-4-(2-methoxy phenyl)-piperazine, has a high affinity (Ki=3.85 nM) for human dopamine D3 receptor with about 70-fold selectivity over the human dopamine D(2L) receptor (Ki=270 nM). KKHA-761 also showed high affinity for cloned human 5-HT1A receptor (Ki=6.4 nM). KKHA-761 exhibited D3 and 5-HT1A receptor antagonist activities in vitro, reversing dopamine- or 5-HT-mediated stimulation of [35S]GTPrS binding. The in vivo pharmacological profile of KKHA-761 was compared with both typical and atypical antipsychotics including clozapine and haloperidol. Apomorphine induced dopaminergic behavior, cage climbing, in mice was potently blocked by a single administration (i.p.) of KKHA-761 (ID50=4.06 mg/kg) or clozapine (ID50=4.0 mg/kg). Cocaine- or MK-801-induced hyperactivity in animals was markedly inhibited by KKHA-761 or clozapine. In addition, KKHA-761 significantly reversed the disruption of prepulse inhibition (PPI) produced by apomorphine in mice, indicating the antidopaminergic or antipsychotic activity of KKHA-761 in mice. However, KKHA-761 was inactive in the forced swimming behavioral despair model in mice, suggesting lack of antidepressant properties. KKHA-761 attenuated the hypothermia induced by a selective dopamine D3 agonist, 7-OH-DPAT, in mice, whereas clozapine enhanced it. Moderate doses of both KKHA-761 and clozapine did not increase serum prolactin levels in rats. Lower doses of, however, haloperidol significantly increased prolactin secretion. KKHA-761 did not induce cataleptic response up to 20 mg/kg, but significant catalepsy was shown at lower doses of clozapine and haloperidol. Furthermore, KKHA-761 showed a low incidence of rotarod ataxia (TD50=34.4 mg/kg, i.p.) in mice. The present results, therefore, suggest that KKHA-761 is a potent antipsychotic agent with combined dopamine D3 and serotonin 5-HT1A receptors modulation activity, which may further enhance its therapeutic potential for anxiety, psychotic depression, and other related disorders. PMID- 16216325 TI - Evaluation of the water genotoxicity from Santos Estuary (Brazil) in relation to the sediment contamination and effluent discharges. AB - The genotoxic activity of water samples collected in 9 different sites within the area of the Santos estuary was preliminary evaluated, and related to previous data on the genotoxicity of sediments and the contents of PAHs in both water and sediment samples. The liquid discharge of a steel mill (coke plant), known to be mutagenic, was chemically analyzed to determine its PAH content. For the water evaluation we employed the Salmonella/microsome assay with the strains TA98 and TA100 with and without S9 mix in the plate incorporation method. The water was filtered with an AP20 membrane before being extracted with XAD4 at natural and acidic pH. The industrial effluent was filtered in 0.45 microm membranes before being extracted with the liquid/liquid method. Both membranes containing the particulate material were extracted using ultrasonication. PAHs were found associated with the suspended particles present in the industrial effluent in accordance with mutagenicity data previously reported. In relation to the estuarine waters, sites 1 and 5 presented low levels of mutagenic activity only in the filtered water (liquid fraction) extracts. At site 3, both the filtered water and particulate solids presented also low mutagenicity. Results show that the mutagenic activity observed in water could not be directly related to the genotoxic activity and PAHs contents of the bottom sediments. PMID- 16216324 TI - Mechanisms of lysophosphatidic acid-induced increase in intracellular calcium in vascular smooth muscle cells. AB - Although lysophosphatidic acid (LPA) is known to cause an increase in intracellular Ca2+ concentration ([Ca2+]i) in vascular smooth muscle cells (VSMCs), the mechanisms of [Ca2+]i mobilization by LPA are not fully understood. In the present study, the effect of LPA on [Ca2+]i mobilization in cultured A10 VSMCs was examined by Fura-2 fluorescence technique. The expression of LPA receptors was studied by immunostaining. LPA was observed to increase [Ca2+]i in a concentration-dependent manner; this increase was dependent on the concentration of extracellular Ca2+. Both sarcolemmal (SL) Na(+)-Ca2+ exchange inhibitors (amiloride, Ni2+ and KB-R7943) and Na(+)-H+ exchange inhibitor (MIA) as well as SL store-operated Ca2+ channel (SOC) antagonists (SK&F 96365, tyrphostin A9 and gadolinium), unlike SL Ca2+ channel antagonists (verapamil and diltiazem), inhibited the LPA-induced increase in [Ca2+]i. In addition, sarcoplasmic reticulum (SR) Ca2+ channel blocker (ryanodine), SR Ca2+ channel opener (caffeine), SR Ca2+ pump ATPase inhibitor (thapsigargin) and inositol 1,4,5-trisphosphate (InsP3) receptor antagonists (xestospongin and 2 aminoethoxydiphenyl borate) were found to inhibit the LPA-induced Ca2+ mobilization. Furthermore, phospholipase C (PLC) inhibitor (U 73122) and protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate) attenuated the LPA induced increase in [Ca2+]i. These results indicate that Ca2+ mobilization by LPA involves extracellular Ca2+ entry through SL Na(+)-Ca2+ exchanger, Na(+)-H+ exchanger and SL SOCs. In addition, ryanodine-sensitive and InsP(3)-sensitive intracellular Ca2+ pools may be associated with the LPA-induced increase in [Ca2+]i. Furthermore, the LPA-induced [Ca2+]i mobilization in VSMCs seems to be due to the activation of both PLC and PKC. PMID- 16216326 TI - Bio-accumulation of some trace metals in the short-neck clam Paphia malabarica from Mandovi estuary, Goa. AB - Monthly collections of the clam Paphia malabarica were made for period of 1 year from May 2002 to April 2003 from Verem, Mandovi estuary, Goa. Clams were categorised into two groups, 25-35 mm (small size) and 35-40 mm (big size). Significant difference was not observed in the accumulation of metals in the whole soft tissue between the two size groups. Irrespective of size, annual mean, metal content recorded as microg/g dry weight in the whole soft tissue was 3.8, 30.3, 13.5, 36.6 and 105.7, respectively, for Cd, Pb, Cu, Zn and Fe. Of the two toxic elements Cd and Pb, Cd content was almost uniform throughout the year except for a rise in September for the small size and October for the big size. Pb, on the other hand, was low from the beginning of the monsoon and exhibited distinct accumulation from December onwards up to April, May and, to some extent, June. The pattern was similar in both the size groups, the values being higher for the bigger size group. The three essential elements Cu, Fe and Zn exhibit trends similar to one another with peaks in September and December-January in both the size groups. Cadmium accumulation was highest in the mantle and adductor muscle, Lead, in foot, Copper, in digestive gland and gonad, and Zn and Fe, in gills. Correlation coefficient between different metal couplings as tested statistically revealed significant coupling for Zn-Fe (r 0.92) in the bigger size group, the same was observed between Cu-Fe (r 0.62) and Cd-Zn (r 0.94). Seasonal difference in Pb accumulation was highly significant. PMID- 16216328 TI - Mycobacterium tuberculosis in Taiwan. AB - OBJECTIVE: The resurgence of tuberculosis (TB) and the emergence of drug resistance of Mycobacterium tuberculosis (MTB) isolates are of great impact on public health. METHODS: Taiwanese data on disease burden of TB and anti-microbial resistance of MTB identified from Annual Reports of Centre for Disease Control, Department of Health, Taiwan and from peer-reviewed publications from MEDLINE (1995-2004). RESULTS: In Taiwan in 2002, the incidence (per 100,000 population) of tuberculosis was 74.6 and it was higher in aborigines (289.8) and in people living in mountainous regions (256.0). The mortality rate of tuberculosis in Taiwan in 2002 was 5.68 per 100,000 population. Susceptibility data summarized from 1990 to 2002 reports showed primary resistance ranged from 4.7 to 12% for isoniazid, 0.7 to 5.9% for rifampin, 1 to 6% for ethambutol, and 4 to 11% for streptomycin. The overall rates of multidrug-resistant tuberculosis (MDRTB) among new cases and previously treated cases were 1 to 3% and 15 to 46%, respectively. The increasing burden of patients with MDRTB infection, the persistent high rate of mortality, the lack of nationwide surveillance system using the standard methodology to determine the trends and current status of resistance, and the inadequate current TB control infrastructure and training to accomplish the tasks required to implement the directly observed treatment short-course (DOTS) strategy are having a great impact on public health in Taiwan. CONCLUSIONS: High disease burden of TB and high resistance rates in MTB as well as inappropriateness of the current control infrastructure for TB services illustrate increasingly serious health problems from TB in Taiwan. PMID- 16216329 TI - Cognitive outcome in adults with moderate disability after pneumococcal meningitis. AB - OBJECTIVES: To assess cognitive outcome and quality of life in patients with moderate disability after bacterial meningitis as compared to patients with good recovery. METHODS: Neuropsychological evaluation was performed in 40 adults after pneumococcal meningitis; 20 patients with moderate disability at discharge on the glasgow outcome scale (GOS score 4) and 20 with good recovery (GOS score 5). RESULTS: Patients with GOS score 4 had similar test results as compared to patients with GOS score 5 for the neuropsychological domains 'intelligence', 'memory' and 'attention and executive functioning'. Patients with GOS score 4 showed less cognitive slowness than patients with GOS score 5. In a linear regression analysis cognitive speed was related to current intelligence, years of education and time since meningitis. Overall performance on the speed composite score correlated significantly with time since meningitis (-0.62; P<0.001). Therefore, difference between both groups may have been related to a longer time between meningitis and testing for GOS four patients (29 vs. 12 months; P<0.001). CONCLUSIONS: Patients with moderate disability after bacterial meningitis are not at higher risk for neuropsychological abnormalities than patients with good recovery. In addition, cognitive slowness after bacterial meningitis may be reversible in time. PMID- 16216330 TI - Recurrent post-sternotomy mediastinitis. AB - The use of muscle flaps or omentum flap combined with an appropriate anti microbial regimen is the treatment of choice for patients with recurrent mediastinitis. Flap closure obliterates the large mediastinal wounds and prevents spreading of the infection to the aorta, heart, including the vascular coronary artery grafts, and other prosthetic materials. In addition, the use of muscle flaps or omentum flap provides the infected tissues with the essential blood supply including good concentration of intravenously administered antibiotics, and leads to early wound closure with sufficient chest stability and respiratory function. PMID- 16216327 TI - Antagonism of HIV-specific CD4+ T cells by C-terminal truncation of a minimum epitope. AB - Antagonism of T cell responses by variants of the cognate peptide is a potential mechanism of viral escape from immune responses and may play a role in the ability of HIV to evade immune control. We show here a rarely described mechanism of antagonism by a peptide shorter than the minimum length epitope for an HIV p24 specific CD4+ T cell clone. The shorter antagonist peptide-MHC complex bound the T cell receptor (TCR), albeit with lower affinity than the full-length agonist peptide. Prior work showing the crystal structure of the peptide-MHC complex revealed a unique glycine hinge near the C-terminus of the agonist peptide, allowing the generation of full-length antagonist peptide lacking the hinge. These results confirm the dependence of productive TCR engagement on residues spilling out from the C-terminus of the MHC binding groove and show that partial engagement of the TCR with a truncated, low-affinity ligand can result in T cell antagonism. PMID- 16216331 TI - Alkhurma hemorrhagic fever virus is an emerging tick-borne flavivirus. PMID- 16216332 TI - Apoptosis in perinatal hypoxic-ischemic brain injury: how important is it and should it be inhibited? AB - The discovery of safe and effective therapies for perinatal hypoxia-ischemia (HI) and stroke remains an unmet goal of perinatal medicine. Hypothermia and antioxidants such as allopurinol are currently under investigation as treatments for neonatal HI. Drugs targeting apoptotic mechanisms are currently being studied in adult diseases such as cancer, stroke, and trauma and have been proposed as potential therapies for perinatal HI and stroke. Before developing antiapoptosis therapies for perinatal brain injury, we must determine whether this form of cell death plays an important role in these injuries and if the inhibition of these pathways promotes more benefit than harm. This review summarizes current evidence for apoptotic mechanisms in perinatal brain injury and addresses issues pertinent to the development of antiapoptosis therapies for perinatal HI and stroke. PMID- 16216333 TI - A model of phototransduction by the human circadian system. AB - The absolute and spectral sensitivities to light by the human circadian system, measured through melatonin suppression or phase shifting response, are beginning to emerge after a quarter century of active research. The present paper outlines a hypothesized model of human circadian phototransduction that is consistent with the known neuroanatomy and physiology of the human visual and circadian systems. Spectral opponency is fundamental to the model, providing a parsimonious explanation of some recently published data. The proposed model offers a framework for hypothesis testing and subsequent discussion of the practical aspects of architectural lighting with respect to light and health. PMID- 16216334 TI - A purposely designed neural signal amplifier for short interval stimulation and recording microneurography using a common electrode. AB - 'Common Electrode' microneurography (i.e. stimulating a nerve and recording return afferent neural activity through the same microelectrode facilitates investigation of linkages between sensory and motor nervous systems in humans. Currently there is no commercial product designed specifically to conduct common electrode microneurography experiments. However, such experiments would advance investigations in several key areas including spinal injury research. In this paper, we report on the successful production and testing (on a human subject) of an integrated amplifier built specifically for this purpose. The amplifier was built using commercially available components to allow for both easy and economical manufacture. In particular, we report on the design requirements and outline our chosen design solutions. The amplifier handles low-level neural signals amidst large 50 Hz interference, with protection against potentially high stimulation voltages of over 100 V dc, with minimal cross-coupling of rapid stimulus pulses onto the high gain amplifier's input, and a short 'blocking' time between stimulation and recording. The amplifier also includes necessary filters, selectable gains and internal stimulator triggering circuits to provide a simple, integrated solution for common electrode operation on human subjects. PMID- 16216335 TI - Search for a two-input model for future investigations of 'synaptic tagging' in freely moving animals in vivo. AB - Processes of "synaptic tagging" guarantee synaptic input specificity after the induction of a protein synthesis-dependent late long-term potentiation (late LTP). Distinct high-frequency stimulation can set a transient "synaptic tag" at the activated synapses, which captures plasticity-related proteins (PRPs) synthesized synapse-non-specifically in dendritic branches/compartments or the somata. Thus, only those synapses, which expressed a "tag", are also able to express late-LTP. Additionally, it was shown that the synthesis of PRPs is triggered by heterosynaptic, non-glutamatergic requirements during LTP-induction in tissue from adult animals. All these experiments were performed in hippocampal slices in vitro so far. Two questions now arise: first, is it possible to describe processes of 'synaptic tagging' in the intact, freely moving animal and second, is the stimulation of glutamatergic inputs sufficient to induce 'tagging' or is the co-activation of a modulatory-heterosynaptic input, also required for the process? We have first developed a technique, which allows us now to induce distinct forms of LTP at the ipsilateral CA1 site by specifically stimulating glutamatergic hippocampal structures at the contralateral site in the intact, freely moving rat. Thus, the used stimulation protocol allowed us to activate two separate synaptic inputs to the same neuronal stimulation, a pre-requisite for tagging-experiments to be investigated in vivo. PMID- 16216337 TI - Characterization and expression analysis of TNF-related apoptosis inducing ligand (TRAIL) in grass carp Ctenopharyngodon idella. AB - TRAIL (Apo2 ligand) described as a type II transmembrane protein belonging to the TNF superfamily can induce apoptotic cell death in a variety of cell types. In the present study, a putative cDNA sequence encoding the 299 amino acids of TRAIL (GC-TRAIL) and its genomic organization were identified in grass carp Ctenopharyngodon idella. The predicted GC-TRAIL sequence showed 44 and 41% identities to chicken and human TRAILs, respectively. In a domain search, a tumor necrosis factor homology domain (THD) was identified in the C-terminal portion of TRAILs. The GC-TRAIL gene consists of five exons, with four intervening introns, spaced over approximately 4 kb of genomic sequence. Analysis of GC-TRAIL promoter region revealed the presence of a number of putative transcription factor binding sites, such as Sp1, NF-kappaB, AP-1, GATA, NFAT, HNF, STAT, P53 and IRF1 sequences which are important for the expression of other TNF family members. Phylogenetic analysis placed GC-TRAIL and the putative zebrafish (Danio rerio) TRAIL obtained from searching the zebrafish database into one separate cluster near mammalian TRAIL genes, but apart from the reported zebrafish TRAIL-like protein, indicating that the GC-TRAIL is an authentic fish TRAIL. Expression analysis revealed that GC-TRAIL is expressed in many tissues, such as in gills, liver, trunk kidney, head kidney, intestine and spleen. PMID- 16216336 TI - Evidence for dissociation of TLR mRNA expression and TLR agonist-mediated functions in bovine macrophages. AB - Toll-like receptors are of key importance in the recognition of and response to infectious agents by cells of the innate immune system. TLR mRNA expression and TLR-mediated functions were determined in bovine macrophages (MPhi) infected with bovine viral diarrhea virus (BVDV) or stimulated with interferon-gamma (IFN gamma) in order to see whether they are correlated under these conditions. As parameters quantitative real time RT-PCR (QRT-PCR) for TLR2, TLR3 and TLR4, NO and TNF production were measured. Triggering of bovine MPhi with bona fide TLR2 and TLR4 agonists (lipopolysaccharide, lipoteichoic acid, peptidoglycan, lipopetide) led to NO and TNF production but neither TLR3 nor TLR9 agonists (double-stranded RNA, CpG DNA) showed this effect. The mRNA expression of TLR2, TLR3 and TLR4 was neither influenced by MPhi costimulation with IFN-gamma nor by MPhi preinfection with BVDV nor by the ligands themselves. However, NO production induced by TLR2 or TLR4 agonists was strongly modulated either by IFN-gamma costimulation or BVDV preinfection. Thus costimulation of MPhi with IFN-gamma resulted in an increase of both NO synthesis and TNF expression by cells stimulated simultaneously by TLR2 or TLR4 agonists. Preinfection of bovine MPhi by BVDV resulted in upregulation of TLR2- and TLR4-mediated NO synthesis. Collectively, these data show that TLR-mediated functions may be modulated by viral infection or activation via IFN-gamma of MPhi whereas the mRNA concentrations of relevant TLR members were not significantly influenced. Thus, the amount of TLR2, TLR3 and TLR4 mRNA transcripts is stable at least under the conditions tested. More importantly, modulation of TLR-mediated responses was dissociated from mRNA expression of TLR members. PMID- 16216338 TI - Cloning and expression of the extra-cellular part of the alpha chain of the equine high-affinity IgE receptor and its use in the detection of IgE. AB - The high-affinity receptor for IgE (FcepsilonRI) plays a central role in IgE mediated allergic reactions. Cross-linking of FcepsilonRI by IgE-antigen complexes results in the activation of mast cells and basophils and is thought to contribute to the immunopathology of Heaves, a chronic obstructive pulmonary disease of horses. Recombinant protein corresponding to the extra-cellular portion of the FcepsilonRI alpha subunit, cloned and sequenced previously, was expressed using both mammalian cells and insect cells. The yield of expressed protein was considerably greater using insect cells and the baculovirus expression system. The recombinant proteins differed in size between the two systems, presumably due to differences in the extent of glycosylation. However, recombinant protein from both cell systems bound equine IgE present in bronchoalveolar lavage fluid from horses with Heaves. These results suggest that the recombinant extra-cellular part of FcepsilonRI should be a useful tool with which to study equine IgE responses. PMID- 16216339 TI - TLR-2 is upregulated and mobilized to the hepatocyte plasma membrane in the space of Disse and to the Kupffer cells TLR-4 dependently during acute endotoxemia in mice. AB - Membrane components of bacteria and fungi are recognized by Toll-like receptors (TLRs) which, when activated, induce several inflammatory mediators important in the host defense. As the liver is constantly exposed to ingested bacteria, hepatic TLRs must be broadly responsive and highly regulated to prevent uncontrolled inflammatory activation. Although several hepatic cells express microbe recognition molecules and inflammatory mediators in vitro, the regulation and cellular localization of these proteins in vivo remain uncertain. The expression and regulation of TLR-2 and TLR-4, and the cytokine expression patterns were evaluated in mouse tissues using a model of acute inflammation induced by intraperitoneal injection of LPS. Five hours after intraperitoneal LPS, induction of TLR-4 was evident in lung, while the low hepatic TLR-4 expression was non-inducible. TLR-2 mRNA and protein were induced both in lung and liver TLR-4 dependently. However, IL-1alpha also contributed to this induction, and IL-1R1 antibody attenuated the TLR-2 increase. Immunoelectron microscopy showed accumulation of cytoplasmic TLR-2 to vesicles near the hepatocyte plasma membrane in the space of Disse, to the sinusoidal endothelium and to the Kupffer cells. NF-kappaB activation was clear in Kupffer cells and hepatocytes during LPS-challenge, suggesting these cells to be the main source of in vivo cytokine production. Hepatic cytokine response to LPS was remarkably rapid in liver, whereas lung responded less acutely. Secondary inflammatory challenge attenuated the TLR-2 response. The innate immune system of the liver is rapidly and transiently activated during endotoxemia by mechanism involving both TLR-4 and TLR-2. PMID- 16216340 TI - Thyroid alar cartilage graft in paediatric laryngotracheal reconstruction. AB - OBJECTIVE: To evaluate the potential indications of thyroid alar cartilage (TAC) graft in the paediatric laryngotracheal reconstruction (LTR) population based on observations obtained in a case series of 27 consecutive infants referred to our tertiary care center. METHODS: Thyroid alar cartilage grafting was performed for limited Myer grade II and grade III subglottic stenosis requiring a single-stage laryngoplasty and for laryngeal enlargement after translaryngotracheal resection of endolaryngeal tumors. The evolution of the grafted area was evaluated prospectively during endoscopic follow-up. RESULTS: Twenty-five patients (92.5%) were successfully extubated after a mean of 5.1 days. No perioperative or postoperative complications were observed. The mean duration of graft harvesting was 7.7 min. Follow-up of the grafted area revealed one case of partial necrosis without prolapse into the lumen. The mean duration of graft epithelialization was 18.1 days (range: 12-30 days). Development of granulation tissue was observed in eight patients (32%) with a mean duration of granulation tissue persistence of 61.5 days (range: 7-155 days). Endoscopic follow-up did not demonstrate any pharyngolaryngeal asymmetry or feeding difficulties. CONCLUSION: This study demonstrated that the use of thyroid alar cartilage grafting is feasible for pediatric laryngotracheal reconstruction. The indications of thyroid alar cartilage graft should be reserved for moderated subglottic stenosis. The use of TAC reduced the operative time and cosmetic sequelae significantly. The healing of the grafted area was similar to those obtained with other types of graft. The TAC removal did not induce laryngeal deformation but longer follow-up is necessary to confirm this. PMID- 16216341 TI - Is fungal infestation of paranasal sinuses more aggressive in pediatric population? AB - OBJECTIVE: Comparison of characteristic features, radiology, management and recurrence pattern of fungal sinusitis between children and adults. METHODS: A prospective study conducted in the department of Otorhinolaryngology, Head and Neck Surgery, Post Graduate Institute of Medical Education and Research, Chandigarh in which all the cases presenting with the features of allergic fungal sinusitis (AFS) between January 2000 and January 2005 were enrolled. These cases were divided into two groups, group 1 comprised of cases with age less than 15 years and group 2 with age more than 15 years. Detailed history, physical examination and nasal endoscopic examination and computed tomography (CT) scan of the paranasal sinuses was done in all the cases. The cases with prior history of sinonasal surgery were excluded from the study. All patients refractory to medical management were subjected to endoscopic sinus surgery. All the cases were followed up for a period ranging from 6 to 39 months to see for the recurrence. The data of both the groups was analysed statistically using chi square test. RESULTS: The study population comprised of 200 cases, with 68 cases in group 1 and 132 cases in group 2. The most common symptom in both the groups was presence of nasal obstruction. The children had higher incidence of having unilateral disease (46 out of 68) compared with adults, where it was 38 out of 132. The bony erosion was seen more often in group 1. Surgery was done endoscopically in all the cases. The intra orbital or intra cranial extension was seen in 58 cases of group 1 and 47 cases of group 2 (p<0.001). Recurrence was seen in 18 (15 with intraorbital and 3 with intracranial extension) cases in group 1 and 13 cases (11 with intraorbital and 2 with intracranial extension) in group 2 (p<0.005). CONCLUSION: In our study, we found a higher incidence of facial deformities, proptosis, intraorbital/intracranial extension and a higher rate of recurrence in group 1, therefore, suggesting a more aggressive nature of AFS in children than adults mandating an early diagnosis, proper management and regular follow up in these cases. PMID- 16216342 TI - Neurocognitive abilities in children with adenotonsillar hypertrophy. AB - OBJECTIVE: Sleep apnea is one of the most deleterious disorders in children with adenotonsillar hypertrophy because it can induce hypoxemia of brain. Sleep apnea may lead to failure to thrive or to physical and mental delay in development, including cognitive disturbances. The aim of this study was to analyze the influence of adenotonsillar hypertrophy, causing obstructive sleep apnea on neurocognitive abnormalities. We were interested in sensorimotor coordination, perception, memory, learning ability, concentration, focused attention and language reception. MATERIALS AND METHODS: We examined 221 children. One-hundred and seventeen children had sleep apnea caused by adenotonsillar hypertrophy: 87 children aged 6-9 years and 34 children aged 10-13 years. The control group, without adenotonsillar hypertrophy, consisted of 104 healthy children. Both groups of children with and without apnea were examined psychologically to determine abilities and minor neurocognitive deficit. The token test (TT), diagnosis test of brain dysfunction (DCS-test), Luria auditory verbal learning test (LAVLT) and Rey complex figure test (RCFT) were applied to both groups. The tests: TT, DCS-test, RCFT were used to investigate the level of sensorimotor integration and perception processes. Memory and learning abilities were measured using LAVLT. The TT assessed language dysfunction in children. RESULTS: This study shows that adenotonsillar hypertrophy in children aged 6-9 years is associated with neurocognitive abnormalities such as: memory problems, concentration of attention deficits, learning disability, language dysfunction, lower sensorimotor integration and perception. The older children (aged 10-13 years) with adenotonsillar hypertrophy had memory problems and learning disabilities. They are likely to be caused by of concentration of attention deficits. The older children were found to have more severe language dysfunction. CONCLUSIONS: Our study shows that sleep apnea may lead to neurocognitive deficits. The results may be helpful in the process of making decision for or against surgery in doubtful cases. It is important to know the problems connected with apnea in order to recognize them and help the child develop by providing adequate treatment and cognitive stimulation. PMID- 16216344 TI - A RT/PCR-partial restriction enzymatic mapping (PREM) method for the molecular characterisation of the large satellite RNAs of Arabis mosaic virus isolates. AB - The satellite RNA of the grapevine isolate NW of Arabis mosaic virus (ArMV) was cloned and sequenced, and showed 75% identity at the nucleotide level to the satellite RNA of the lilac isolate of ArMV. In order to survey ArMV isolates from various geographical origins and natural hosts for the presence of large satellite RNAs and analyse their degree of variability, a RT/PCR-partial restriction enzymatic mapping (PREM) method was developed. The method is based on the incorporation of 5-methyl-dCTP in the RT/PCR reaction, and the subsequent digestion of the RT/PCR products by methyl-sensitive restriction enzymes. Satellites RNAs were detected by RT/PCR in eight isolates out of 47, six of them originating from grapevine, one from hop and one from lilac. The partial restriction digestion patterns allowed to distinguish six different types of satellites. Cloning and sequencing of the different satellites confirmed these results, the PREM proving able to discriminate sequences with 96% identity. The sizes of the different satellites varied between 1092 and 1139 nucleotides, their encoded proteins between 338 and 360 amino acids. Conserved domains were found in the amino and carboxy-termini between the sequences of the proteins encoded by the satellites of the different isolates of ArMV. PMID- 16216345 TI - Molecular basis of programmed cell death involved in neurodegeneration. AB - Rapid progress in understanding the molecular basis of neurodegeneration has been tightly linked with recent discoveries in the field of programmed cell death (PCD). Analysis of PCD in neuronal demise has led to identification of several associated phenomena, such as re-initiation of the cell cycle and the key role of oxidative stress, although putative causal relationships between these events are still debatable. These issues are reviewed here in the context of acute and chronic neurodegenerative processes. In addition, newly emerging concepts concerning cell-cycle re-initiation are discussed in terms of their potential impact on the development of more effective therapeutic strategies. PMID- 16216346 TI - Sounds do-able: auditory-motor transformations and the posterior temporal plane. AB - Accumulating evidence in humans and non-human primates implicates the posterior superior temporal plane (STP) in the processing of both auditory spatial information and vocal sounds. Such evidence is difficult to reconcile with existing accounts of the primate auditory brain. We propose that the posteromedial STP generates sequenced auditory representations by matching incoming auditory information with stored templates. These sequenced auditory representations are subsequently used to constrain motor responses. We argue for a re-assessment of the much-debated dorsal auditory pathway in terms of its generic behavioral role as an auditory "do" pathway. PMID- 16216347 TI - A single session of emotional stress produces anxiety in Wistar rats. AB - The sensorial stimulation arising from a physically stressed (PS) subject may produce emotional stress in a witnessing partner (WP). Both members of the pair develop functional changes. We tested changes in locomotor activity (crossing) and in the defensive burying test in WP, and PS adult male Wistar rats having been submitted to a single 10 min session in a two-compartment cage. During this session, the WP rats received auditory and olfactory stimulation coming from a PS pair submitted to unavoidable electric footshocks (1 mA, dc, 0.5s, 0.5c/s, 10 min). This experiment was replicated in other groups pre-treated with vehicle or diazepam, and their urine was collected and analyzed by the static Head-Space and Gas Chromatography-Mass Spectrometry (HS-GC/MS) techniques. The WP group displayed a significantly higher crossing [F((2,45))=4.31, P<0.01] and more cumulative burying time [F((2,22))=4.73, P<0.01] than the control or PS groups. Diazepam (1mg/kg) reverted these changes. Our results indicate that the conspecific sensorial communication coming from the PS group produces anxiety probably mediated by 2-heptanone, since the HS-GC/MS analyses showed the highest amount of 2-heptanone in the urine from the PS group [F((2,42))=5.17, P<0.009]. PMID- 16216348 TI - Social and spatial changes induce multiple survival regimes for new neurons in two regions of the adult brain: An anatomical representation of time? AB - Male zebra finches reared in family groups were housed initially in small indoors cages with three other companions. At 4-5 months of age these birds were treated with [(3)H]-thymidine and then placed in large outdoors aviaries by themselves or with other zebra finches. Counts of new neurons were made 40, 60 and 150 days after the change in housing. Recruitment of new neurons in nidopallium caudale (NC) was higher than in the hippocampal complex (HC); but in both brain regions it was higher in communally housed birds than in birds housed singly, suggesting that the complexity of the social setting affects new neuron survival. In addition, the new neurons lived longer in rostral NC than in its caudal counterpart, and neuronal turnover was faster and more significant in NC than in HC. Albeit indirect, this may be the first suggestion that different parts of the brain upgrade memories at different time intervals, yielding an anatomical representation of time. PMID- 16216349 TI - Humic acid reduces gonadotropin activity and hormonal sensitivity of frog oocytes. AB - The specific stimulatory effect of sturgeon Acipenser guldenstadti Br. gonadotropic hormone (GTH) on frog Rana temporaria L. oocyte maturation in vitro was investigated in relation to humic acid (HA) concentrations from 12.5 to 50 mg/l. HA was observed to bind to both the follicular membrane of the oocytes and the GTH molecule, reducing the oocytes' hormone sensitivity and maturation ability. It was also shown that HA inactivated GTH, lowering its specific ability to stimulate oocyte maturation. PMID- 16216350 TI - Regulation of inhibitor of differentiation gene 3 (Id3) expression by Sp2-motif binding factor in myogenic C2C12 cells: downregulation of DNA binding activity following skeletal muscle differentiation. AB - Id3 is a member of the Id family of transcriptional regulators that have been implicated in the development of multiple tissues. Altered expression of the Id genes and proteins contribute to carcinogenesis and atherosclerosis. Id3 is highly expressed in proliferating skeletal muscle cells but becomes downregulated upon terminal differentiation. We have identified several DNase I protected footprints within a proximal region of the mouse Id3 promoter that has been shown previously to support high levels of transcriptional activity in proliferating skeletal muscle cells. Two of these sites interacted, respectively, in vitro with Sp2 and Egr-1 proteins present in muscle cell nuclear extracts. Mutation analysis revealed that the Sp2 site accounted for a major part of the Id3 promoter activity in proliferating muscle cells whereas the Egr-1 site was dispensable. Consistent with the previously observed downregulation of the endogenous Id3 gene, protein binding to the Sp2 site was substantially reduced with extracts from differentiated muscle cells. Our results reveal Id3 as a potential target for Sp2 and raise the possibility that acute activation and the chronic and maintained expression of Id3 gene might be regulated by different mechanisms. PMID- 16216351 TI - Predictors of non-pharmacological intervention in patients with paroxysmal atrial fibrillation: Value of neuroticism. AB - BACKGROUND: Non-pharmacological intervention is gaining increasing popularity in the treatment of patients with paroxysmal atrial fibrillation. We sought to investigate which factors play a role in the choice for non-pharmacological intervention with a particular focus on neuroticism. METHODS: The study group comprised 73 patients with paroxysmal atrial fibrillation (mean age 55+/-13 years, 50 males). On average, patients had a 3-year-history of one symptomatic paroxysm per week lasting 2 h. The degree of neuroticism was assessed using the short scale Eysenck Personality Questionnaire. RESULTS: During a mean follow-up period of 7.0+/-0.6 years, 20 patients (27%) underwent a non-pharmacological intervention for atrial fibrillation including His bundle ablation (n = 1), maze operation (n = 4), DDDR-pacemaker (n = 10), pulmonary vein ablation (n = 5). Multivariate regression analysis showed that age < 55 years (odds ratio 5.3, 95% CI 1.1-24.5), frequency of paroxysms of atrial fibrillation > 1 per week (odds ratio 5.9, 95% CI 1.2-28.5) and total number of anti-arrhythmic drugs (class I and III) used > 2 (odds ratio 3.4, 95% CI 1.6-6.9) were predictive of non pharmacological intervention (all p < 0.05). In contrast, the degree of neuroticism was similar in patients who underwent non-pharmacological intervention as opposed to patients who did not undergo non-pharmacological intervention (4.5+/-3.3 vs. 4.0+/-2.9, p = NS). CONCLUSIONS: On the basis of this small study, neuroticism would not appear to play an important role in the decision to perform a non-pharmacological intervention. Instead, the data indicate that younger patients with pharmacologically refractory atrial fibrillation more often undergo non-pharmacological intervention. PMID- 16216352 TI - Cytokines in acute coronary syndromes. AB - The aim of this study was to show the presence of an imbalance between pro inflammatory and anti-inflammatory mediators in patients affected by acute coronary syndromes (ACS). We evaluated the production in cultured and stimulated lymphomonocytes of interferon (IFN)gamma and tumor necrosis factor (TNF)alpha, which are well known to possess pro-inflammatory effects, and of interleukin (IL)10, which has been shown to have a protective anti-inflammatory activity, in two groups of 30 patients affected by acute myocardial infarction (AMI) and unstable angina (UA), compared with two equivalent groups of patients with stable angina (SA) and of healthy volunteers. We found a significant increase of IFNgamma and TNFalpha production (p<0.01) and a significant decrease of IL-10 production (p<0.01) in cultures of lymphomonocytes taken from patients with AMI and UA compared with SA patients and controls. No significant changes were found between AMI and UA patients and SA patients and controls. We conclude that a relevant imbalance in cytokine release is present in ACS, markedly favoring pro inflammatory effects. PMID- 16216353 TI - Estimated BVDV-prevalence, -contact and -vaccine use in dairy herds in Northern Portugal. AB - A study to evaluate BVDV-prevalence, recent -contact and -vaccine use in dairy herds in the "Entre Douro e Minho" (EDM) region in North Portugal was carried out in 124 dairy herds in 2003. Herds were visited to ascertain BVDV-vaccine use and to collect a bulk tank milk (BTM) sample and serum from 1268 cattle to analyse BVDV-antibodies using an NS2-3 ELISA. Fifty-three percent of farmers used inactivated BVDV-vaccines whilst the remaining farmers were not presently using BVDV-vaccines. BMT-antibody results included 35% positives, 25% negative and 39% inconclusive, and were similar in vaccinated and non-vaccinated herds (p>0.05) and allowed estimating a 10% BVDV herd-prevalence from prior knowledge of the relationship between BMT-antibody results and probability of PI cattle in the herd. Overall individual seroprevalence was 27% and was 23% in non-vaccinated and 36% in vaccinated cattle (p<0.05). Contact of the herd with BVDV was assessed according to seroprevalence in young and adult cattle in the herd and it was estimated that 35% of herds were infected or had recent contact with BVDV, 40% were not infected and did not have recent contact with BVDV and the BVDV infection and -contact status of remaining herds was undetermined. The results from this study indicate BVDV is endemic and BVDV-vaccines are widespread in the dairy-cattle population in EDM region in Portugal. PMID- 16216354 TI - The copy-number of plasmids and other genetic elements can be determined by SYBR Green-based quantitative real-time PCR. AB - In this study, we explored whether SYBR Green-based quantitative real-time PCR (qPCR) could be used to determine the copy number of a plasmid and whether the method was broadly applicable to chromosomally encoded genetic elements often occurring in multiple copies, such as rRNA genes and insertion sequences (IS). Three different template sources (whole cells, total DNA, and restriction-enzyme digested total DNA) derived from the bacterium Comamonas sp. strain JS46 were analyzed by qPCR using primer-pairs targeting plasmid pJS46 and three chromosomally encoded sequences (16S rDNA, ISCsp1, and IS1071). The difference between threshold cycle values, C(T), of amplicons targeting these elements and of an amplicon targeting the single-copy reference element mnbA (chromosomally encoded) was used to establish DeltaC(T). DeltaC(T) values were then used to derive copy number. For pJS46, qPCR analyses of whole cells and total DNA underestimated the copy number of pJS46 approximately 7-fold and approximately 2.5-fold, respectively, whereas copy number values derived from qPCR analyses of digested total DNA were comparable to those derived from Southern blot (SB) analyses. In contrast, for the chromosomally encoded elements, qPCR analyses of all three template sources gave copy number values that were virtually identical to or differed by approximately 2 from copy number values derived by SB analysis. These data indicate that qPCR can be used to estimate the copy number of various genetic elements but that the accuracy of qPCR-derived values is affected by the template source. PMID- 16216355 TI - A CARD-FISH protocol for the identification and enumeration of epiphytic bacteria on marine algae. AB - A CARD-FISH protocol was developed and applied to analyse surface-associated bacteria on the marine algae Ulva lactuca, Delisea pulchra, Corallina officinalis, Amphiroa anceps, Porphyra sp. and Sargassum linearifolium. The combination of Alexa(546)-labelled tyramide as the reporter molecule with SYBR Green II counterstain allowed for superior detection of the hybridised probe fluorescence against plant tissue from which pigment autofluorescence has been reduced. PMID- 16216356 TI - Rapid bacterial identification using evanescent-waveguide oligonucleotide microarray classification. AB - Bacterial identification relies primarily on culture-based methodologies and requires 48-72 h to deliver results. We developed and used i) a bioinformatics strategy to select oligonucleotide signature probes, ii) a rapid procedure for RNA labelling and hybridization, iii) an evanescent-waveguide oligoarray with exquisite signal/noise performance, and iv) informatics methods for microarray data analysis. Unique 19-mer signature oligonucleotides were selected in the 5' end of 16s rDNA genes of human pathogenic bacteria. Oligonucleotides spotted onto a Ta(2)O(5)-coated microarray surface were incubated with chemically labelled total bacterial RNA. Rapid hybridization and stringent washings were performed before scanning and analyzing the slide. In the present paper, the eight most abundant bacterial pathogens representing >54% of positive blood cultures were selected. Hierarchical clustering analysis of hybridization data revealed characteristic patterns, even for closely related species. We then evaluated artificial intelligence-based approaches that outperformed conventional threshold based identification schemes on cognate probes. At this stage, the complete procedure applied to spiked blood cultures was completed in less than 6 h. In conclusion, when coupled to optimal signal detection strategy, microarrays provide bacterial identification within a few hours post-sampling, allowing targeted antimicrobial prescription. PMID- 16216357 TI - Hematopoietic stem cell gene therapy: dead or alive? AB - Despite some reports of toxicity in recent clinical trials, many scientists believe that the use of gene therapy in the treatment of congenital genetic defects and acquired disorders has too much potential to abandon. Hematopoietic stem cells (HSCs) have been primary targets for gene therapy owing to their capacity for differentiation and self-renewal, whereby multiple cell lineages can potentially be corrected for the lifetime of an individual. These efforts represent a long-term investment towards broadening physicians' treatment options for patients whose diseases, in particular certain immunodeficiencies, are fatal and where no other therapy is available. We review the recent progress and clinical triumphs as well as the reported toxicity related to insertional mutagenesis. We also discuss the current risk-to-benefit estimates and future strategies to reduce the risks and allow full realization of clinical potential. Scientists are continually revising protocols: going both from "bench to bedside" and, as strikingly demonstrated by HSC gene therapy, from "bedside to bench." PMID- 16216358 TI - Weak protein-protein interactions as probed by NMR spectroscopy. AB - Weak protein-protein interactions (PPIs) are fundamental to many cellular processes, such as reversible cell-cell contact, rapid enzyme turnover and transient assembly and/or reassembly of large signaling complexes. However, structural and functional characterizations of weak PPIs have been technically challenging and lagged behind those for strong PPIs. Here, we describe nuclear magnetic resonance (NMR) spectroscopy as a highly effective tool for unraveling the atomic details of weak PPIs. We highlight the recent advances of how NMR can be used to rapidly detect and structurally determine extremely weak PPIs (K(d)>10(-4)M). Coupled with functional approaches, NMR has the potential to look into a wide variety of biologically important weak PPIs at the detailed molecular level, thereby facilitating a thorough view of how proteins function in living cells. PMID- 16216359 TI - Robust treatment planning for intensity modulated radiotherapy of prostate cancer based on coverage probabilities. AB - BACKGROUND AND PURPOSE: To evaluate an optimization approach where coverage probabilities are incorporated into the optimization of intensity modulated radiotherapy (IMRT) to overcome the problem of margin definition in the case of overlapping planning target volume and organs at risk. PATIENTS AND METHODS: IMRT plans were generated for three optimization approaches: based on a planning CT plus margin (A), on prostate and rectum contours from five pre-treatment CT plus margin (B), and on coverage probabilities (C). For approach (C), the probability of organ occupation was computed for each voxel from five pre-treatment CTs and the population distribution of systematic setup error and it was used as local weight in the costfunctions. Monte Carlo simulations of treatment courses were used to compute the probability distribution of prostate and rectal wall equivalent uniform dose (EUD). RESULTS: Treatment simulations showed best and most robust results for prostate and rectal wall EUD within the population for (C). For (A) the rectal wall EUD was on average about 1.5 Gy greater than in (C), while the prostate EUD was lower than those from (C) for most of the patients for (B) (especially for those with great organ motion). CONCLUSIONS: The incorporation of coverage probabilities as local weights allows for dose escalation as well as improved rectal sparing and results in a safer and more robust IMRT treatment. PMID- 16216360 TI - A phase II trial of low-dose total body irradiation and subcutaneous interleukin 2 in metastatic melanoma. AB - BACKGROUND AND PURPOSE: Our own experimental data suggests a therapeutic synergism between low-dose total body irradiation (LTBI) and interleukin-2 (IL 2). PATIENTS AND METHODS: Forty-five patients received a maximum of 2 cycles of high dose subcutaneous (s.c.) IL-2 and LTBI. One treatment cycle included 5 weeks treatment followed by 2 weeks break and composed of a single radiation fraction 0.1 Gy on days 1, 8, 22 and 30 and IL-2: 18 MU x 2 daily s.c. on days 2 to 5 and days 16-19 as well as 9 MU x 2 daily s.c. on days 9-12 and 31-34. In 17 patients, flow cytometric analyses of the various subpopulations of immune cells were done on blood samples before the first LTBI fraction and 24h after LTBI as well as after the first week of treatment. RESULTS: Two patients (4.4%) had a partial response (PR) and 13 patients (29%) had stable disease (SD). The duration of the partial remission and stable disease did not exceed 3 months. The median overall survival was 5.8 months (95% CI, 4-8 months). Thirty-four of the 58 treatment cycles (74%) were given in 100% of the intended dose without modification or delay. The dose was modified in 15 cycles (26%) because of progression (6), liver toxicity (3), CNS toxicity (2), thrombocytopenia (1), lung morbidity (1) and itching (1). There were no treatment-related deaths. Flowcytometry data showed a significant increase in the percentage of cells carrying the beta chain of IL-2 receptor (CD122+), a significant increase in the percentage of NK cells (CD56+ cells) as well as a significant reduction in the percentage of B cells (CD20+) and monocytes (CD14+). CONCLUSIONS: This LTBI and IL-2 regimen was well tolerated, however it cannot be recommended because of its low clinical efficacy. No indication of increased efficacy or altered toxicity was seen using LTBI. PMID- 16216361 TI - Experience of external beam radiotherapy given adjuvantly or at relapse following surgery for craniopharyngioma. AB - BACKGROUND AND PURPOSE: Evaluation of effect of timing of external beam radiation therapy (EBRT) following surgery for craniopharyngioma. MATERIALS AND METHODS: Between 1976 and 2002, 87 patients (28 children) received EBRT in a regional referral centre. Forty-four patients received EBRT adjuvantly and 43 on relapse. The median total dose was 42.5Gy (range 34.7-52.5Gy) in 2.25-2.83Gy fractions over a median of 20 days (range 17-32). Effect of EBRT timing, type of original surgery, age on survival, progression-free survival (PFS) and quality of life (QOL) was studied. RESULTS: Survival from diagnosis was 86 and 76% and PFS was 78 and 66% at 10 and 20 years, respectively, with no significant difference seen between those treated adjuvantly or at relapse or according to age. QOL deteriorated significantly from diagnosis to last follow-up. Excluding patients who relapsed following EBRT, QOL did not deteriorate significantly overall (P=0.35). Children had worse QOL and greater morbidity at all timepoints compared to adults. CONCLUSIONS: EBRT is effective both adjuvantly and at relapse. QOL deteriorates over time-relapse following EBRT was the only significant factor. Children have greater morbidity compared to adults, but no evidence for greater EBRT-induced toxicity was seen. PMID- 16216362 TI - Double-blind randomised phase II study of hyperbaric oxygen in patients with radiation-induced brachial plexopathy. PMID- 16216363 TI - Monte Carlo calculations of the dose distribution around a commercial gynecologic tandem applicator. AB - BACKGROUND AND PURPOSE: Dose rate distributions around Fletcher Suit Delclos (FSD) tandem applicators used for intracavitary brachytherapy are usually calculated by assuming each source is a point source and summing the contributions from each of the sources. Consequently, interpellet attenuation and scattering are ignored. Additional error may be introduced because the applicator walls and tip screw are not considered. The focus of this study was a Monte Carlo simulation of a Selectron tandem, verification of the calculations, and presentation of the implications of the point-source approximation for treatment planning. MATERIALS AND METHODS: MCNPX 2.4.k was used to calculate dose rate distributions around straight and curved tandems. The Monte Carlo calculations were verified with radiochromic film. RESULTS: MCNPX calculated dose to within +/ 2% or +/-2 mm for 97% of the points on the film parallel to the long axis and 98% on a film perpendicular to the long axis of the straight portion of the tandem. The point source approximation overestimated dose by as much as 33% superior to the tip of the tandem as compared to MCNPX. The point source approximation overestimated dose when photons passed through multiple pellets by as much as 18% as compared to MCNPX. Laterally, the dose distribution was not affected greatly. CONCLUSIONS: Interpellet attenuation was a dominant factor in determining the distribution along the length of the pellet train. MCNPX calculated doses accurately when the pellets and applicator walls were included in the geometry. The point source approximation is adequate lateral to the tandem. The point source approximation does not calculate dose accurately superior to the tandem or when photons pass through multiple pellets. PMID- 16216364 TI - Primary radical external beam radiotherapy of rectal adenocarcinoma: long term outcome of 271 patients. AB - BACKGROUND AND PURPOSE: To assess local control, survival and toxicity following primary radical external beam radiotherapy of rectal adenocarcinoma treated between 1978 and 1997, and to compare practices and outcomes between the two decades of study. PATIENTS AND METHODS: A retrospective review of 271 patients who underwent primary radical external beam radiotherapy, without prior or concurrent chemotherapy, was conducted. All patients received radiation doses of at least 40 Gy. Most patients (82%) were followed until death. RESULTS: Tumor fixation was the only significant prognostic factor for complete response by multivariate analysis. Complete response rates for mobile, partially fixed and fixed tumors were 49, 22 and 9%, respectively. For patients with mobile, partially fixed and fixed tumors, overall 5 year survival was 48, 26 and 6%, respectively, and cancer specific 5 year survival was 59, 33 and 9%, respectively. On multivariate analysis, tumor fixation and CEA higher than 8.7 microg/L at presentation were associated with significantly shortened survival. Outcomes did not change significantly during the two decades of the study. RTOG grade 3 or higher acute (1.8%) and late (1.5%) toxicity occurred only in the first decade of study. CONCLUSIONS: Radical external beam radiotherapy is a reasonable treatment option for rectal carcinoma for patients who are not surgical candidates or refuse surgery. However in patients with fixed tumors in this study, the outcome is poor and treatment should be considered palliative. Severe radiation toxicity is infrequent with current schedules and techniques of radiotherapy. PMID- 16216365 TI - 192Ir or 125I prostate brachytherapy as a boost to external beam radiotherapy in locally advanced prostatic cancer: a dosimetric point of view. AB - PURPOSE: This work aims at comparing the dosimetric possibilities of 125I or 192Ir prostate brachytherapy (Bt) as a boost to external beam radiotherapy in the treatment of locally advanced adenocarcinoma. METHODS AND MATERIALS: From 1/1997 to 12/2002, 260 patients were treated. Until 12/2001 a low dose rate (LDR) treatment with 192Ir wires was used, later replaced by a high dose rate (HDR) delivered with an 192Ir stepping source technology. For the present work, we selected 40 patients including the last 20 treated, respectively, by LDR and HDR. The planning CT Scans of all these 40 patients were transferred into the 3D Prowess system for 125I permanent implants design according to the Seattle method. The reference data for dosimetric comparisons were the V100 and the prescribed dose for 192Ir as well as the dose delivered with 125I techniques to the 192Ir V100. We compared V100-150 data as well as doses to the organs at risks (OR) and cold spots (CS). RESULTS: The V100 is 85.3+/-8% for 192Ir LDR and 96+/ 2% for HDR techniques (P < 0.0001). In comparison with 125I, the 192Ir LDR mode induces higher hyperdosage volumes inside the CTV but also more CS, while maximal doses to urethra and rectum are, respectively, 17 and 39% less with 125I (P < 0.0001). In comparison with the 192Ir HDR mode, 125I Bt induces higher hyperdosage volumes and slightly more CS deliberately planned around the bladder neck. If delivered doses to urethra are identical, those to the 20% anterior part of the rectum are 33% less with 125I (P < 0,0001). The 125I Bt technique was only possible in 24 out of the 40 patients studied due to pelvic bone arch interference. CONCLUSIONS: At the present time, there is no evident dosimetric superiority of one Bt method when all the criteria are taken into account. However, improving Bt techniques to implant any prostatic size could found the superiority of the 125I or permanent implants. 125I indeed allows large hyperdosage volumes inside the CTV in comparison with 192Ir HDR techniques while lowering doses to OR and minimizing CS. PMID- 16216366 TI - Comparative characterization of Listeria monocytogenes isolated from Portuguese farmhouse ewe's cheese and from humans. AB - In order to investigate the possible relationships between Listeria monocytogenes strains isolated from farmhouse ewe's cheese and clinical strains collected, in partially overlapping dates, from the same geographical area in Portugal, a total of 109 isolates from seven ewe's cheese manufactures (n=94) and from humans (n=15) were characterized by serotyping, RAPD, PFGE and allelic analysis of the virulent actA gene. Serotyping indicated the presence of four different serovars: 1/2a, 1/2b, 1/2c and 4b. The 15 clinical isolates were either serovar 4b (86.7%) or serovar 1/2b (13.3%). Among the 94 isolates from cheese and related environments the serovars prevalence was 1/2a (1.1%), 1/2b (17.0%), 1/2c (12.8%) and, unexpectedly, 4b (69.1%). Based on results obtained with PFGE typing of the strains, 25 genotypes were identified, 10 from farmhouses and 15 from human cases. Isolates from serovars 1/2a and 1/2c were assigned to single genotypes, respectively. Within serovars 1/2b and 4b three and 20 genotypes were established, respectively. RAPD typing of the isolates rendered 18 types indicating the lack of accuracy of the primers used in strain differentiation within serovar 4b. The actA gene typing of the strains showed a prevalence of actA gene type I (90.4%) compared with the rest of the strains that were all actA gene type II (9.6%). In spite of the fact that all the farmhouses were completely independent, the distribution of L. monocytogenes genotypes, intra and inter cheese manufactures, was relatively homogeneous, suggesting the existence of resident strains. In contrast, among human isolates there was a great genetic diversity. There was no common genotype between L. monocytogenes implicated in the cases of listeriosis and these cheese-related isolates, suggesting the absence of a causal relationship. PMID- 16216367 TI - High pressure-induced inactivation of Qbeta coliphage and c2 phage in oysters and in culture media. AB - High pressure (HP) treatment inactivates bacteria in shellfish, but its effects on viruses in shellfish have not yet been determined, although viral illness is frequently associated with shellfish consumption. The aim of this study was to investigate the baroresistance of two bacteriophage viruses, Qbeta coliphage and c2 phage, in oysters and in culture media. High numbers (>or=10(7) ml(-1) or g( 1)) of both phages were obtained in culture media and in oysters. Samples were HP treated at 200-800 MPa at 20 degrees C for up to 30 min. Little or no inactivation of either phage was observed in oysters or in culture media after treatment at 100 degrees F (12.5%). The 12.5% reported missing work because of vaccine adverse events. Most adverse events were anticipated and of short duration. PMID- 16216395 TI - Pathogenicity and protective immunogenicity of cysteine proteinase-deficient mutants of Leishmania mexicana in non-murine models. AB - This study demonstrates that deletion of cysteine proteinase (CP) genes diminishes pathogenicity of Leishmania mexicana in non-murine experimental host models while preserving immunogenicity. Both cpb and cpa/cpb-deficient lines induced delayed disease onset, smaller lesions and lower parasite burden in hamsters. cpa/cpb-deficient L. mexicana grew more slowly as promastigotes and presented lower infectivity and growth in human mononuclear phagocytic host cells. Protection against homologous challenge comparable to that induced by infection with the virulent wild-type (WT) L. mexicana strain was achieved in the highly susceptible hamster model by immunization with 1000 cpb-deficient promastigotes. CP-deficient L. mexicana elicited significantly lower levels of Th2-associated cytokines IL-10 and TGF-beta than the WT in the primary lesion of hamsters. These findings support the feasibility of using genetically attenuated live Leishmania to achieve protective immunity. PMID- 16216397 TI - Is there a need for antibiotic prophylaxis after routine dental extraction in diabetic patients? PMID- 16216398 TI - The effect of turbation on zinc relocation in a vertical floodplain soil profile. AB - Turbation is hypothesized to affect the redistribution of heavy metals in polluted floodplain soils by effects on mobility. This hypothesis was tested in microcosms by turbation of zinc-spiked sediment top layers. Manual turbation caused a fast decrease of the zinc content in the upper 15 cm of the soil, even though turbation was only applied to the upper two centimetres. It was especially zinc attached to colloid and organic matter particles that was redistributed from the top layer. Percolation processes resulted in the attached zinc being drained to depths of more than 15 cm. The decrease in zinc content of the topsoil was even stronger in combination with inundation. No indications were found for the redistribution of zinc as a result of an increase of the extractability with 0.01 M CaCl2 or changes in pH. The findings suggest that mechanical turbation and bioturbation may redistribute heavy metals from topsoils in polluted floodplains just after inundation as observed in these turbation experiments. PMID- 16216399 TI - Export of nutrients from golf courses on the Precambrian Shield. AB - Annual export rates, or fluxes, of total nitrogen (TN), nitrate, total phosphorus (TP) and potassium from four streams on two golf courses on the Precambrian Shield were compared with those from forested reference locations. Overall, the mean annual fluxes of K, TN, NO(3) and TP from golf courses were greater than from forested areas by 10, 2, 6 and 2 times, respectively. The overall mean export coefficients (kg/ha/yr) were 16 for K, 5.2 for TN, 2.1 for NO(3) and 0.14 for TP. For TN and TP, these are similar to those reported from cropland in Canada by Chambers and Dale. PMID- 16216401 TI - Additive or synergetic effects of phenolic compounds on human low density lipoprotein oxidation. AB - The in vitro assessment of the antioxidant capacity of four phenolic compounds; catechin, hesperidin, ferulic acid, and quercetin was evaluated by the examination of their ability to inhibit copper (Cu(2+))-mediated human low density lipoprotein (LDL) oxidation by using the thiobarbituric acid-reactive substances (TBARS) assay. Individually, the enrichment of LDLs with various concentrations of catechin, hesperidin, ferulic acid, and quercetin produced both antioxidant and prooxidant effects depending on enrichment concentrations of the polyphenolic compounds. Catechin and hesperidin had predominantly antioxidant effects (51.1%, 76.9%, respectively) while ferulic acid and quercetin had mostly prooxidant effects (166.4%; 191.8%, respectively) on LDL oxidation. However, when the mixture of the four phenolic compounds was used to enrich the LDLs, significant antioxidant capacity was demonstrated at all enrichment levels with a dose-response. Synergistic effects of the polyphenolic compounds as mixtures in preventing human LDL oxidation may reflect that nutritional advantages are found in the consumption of a variety of fruits and vegetables in preventing LDL oxidation and perhaps a host of cardiovascular diseases. PMID- 16216402 TI - Detection of 2,4-dichlorophenoxyacetic acid in rat milk of dams exposed during lactation and milk analysis of their major components. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D) and its derivatives are herbicides widely used to control the growth of broadleaf and woody plant. Human and animal exposure to 2,4-D through agriculture use, food products, or use in lawn and garden care has been well documented, but little information is available on the transfer from serum to milk in exposed dams. In this study, we measured the content of 2,4-D in rat milk from mother exposed to 15, 25, 50 or 7 0mg 2,4-D/kg bw through the diet (4 treated groups, 8 dam each; 1 control group with 8 dams) over a period of 16 days starting on the post-natal day 1 (PND 1). The effect of 2,4-D on milk components was also evaluated. All doses tested caused a decrease in the body weight gain of the pups (4 groups, 64 pups each). It also produce a 30% in the content of total lipids and a changed the content of minor proteins in milk of the treated groups. 2,4-D produces an important decrease in some fatty acids content, being the polyunsaturated fatty acids the most affected. Further analysis showed that 2,4-D concentrations chromatographically detected both serum of dams and pups and milk were dose-dependent. PMID- 16216400 TI - How positive affect modulates cognitive control: the costs and benefits of reduced maintenance capability. AB - Adaptive action in a constantly changing environment requires the ability to maintain intentions and goals over time and to flexibly switch between these goals in response to significant changes. argued that positive affect modulates these antagonistic control demands in favor of a more flexible but also more distractible behavior. In the present paper, the author will present further evidence for the affective modulation of cognitive control: mild positive affect reduced maintenance capability in a simple cuing paradigm (the AX Continuous Performance Task) as compared to negative and neutral affect. This reduced maintenance capability results in costs when a to be maintained goal has to be executed and conversely results in benefits when a to be maintained goal unexpectedly changes. The data will be discussed with respect to existing theories on positive affect, cognitive control, and dopamine. PMID- 16216403 TI - Levels of polychlorinated biphenyls (PCBs) in meat and meat products from the state of Rio Grande do Sul, Brazil. AB - The levels of six polychlorinated biphenyl congeners (PCBs) were evaluated in 55 samples of meat (bovine and pork) and meat products (sausage, hot dog sausage, bologna sausage, canned export meat and salami) from 11 cities of the state of Rio Grande do Sul, Brazil, between July and August 2002. PCB congeners were found (in fat basis) in the following rank 52 (5.18 ng/g) > 180 (1.69 ng/g) > 10 (1.35 ng/g) > 28 (1.19 ng/g) >153 (0.47 ng/g) >138 (0.43 ng/g), with a summation SigmaPCB amounting to 10.30 ng/g. Meat products had higher PCB levels than meat. PCB levels in samples followed the rank: mixed meat products > pork meat > bovine meat. These results indicate the presence of PCBs in food samples from Rio Grande do Sul, Brazil, but the levels found were well below the maximum level established for animal food products in Brazil (3000 ng/g fat). Only one sample exceeded the maximum level established by the European Community (200 ng/g fat). This is the first paper describing background concentrations of PCBs in meat and meat products from Brazil. PMID- 16216404 TI - An alcohol-free tissue conditioner--a laboratory evaluation. AB - OBJECTIVES: An alcohol-free tissue conditioner based on a n-butyl methacrylate/i butyl methacrylate copolymer has recently been developed. The purpose of the present study was to compare some key properties of the new tissue conditioner with those of poly(ethyl methacrylate)-based conventional materials containing ethyl alcohol. The effect of a coating, which consisted of poly(ethyl methacrylate) and methyl methacrylate, was also evaluated. METHODS: The new alcohol-free tissue conditioner (Fictioner) and three tissue conditioners containing ethyl alcohol (FITT, Hydro-Cast, SR-Ivoseal) were evaluated. The coated alcohol-free material was also used. Gelation characteristics, dynamic viscoelastic properties and compatibility with dental stones were measured using a displacement rheometer, dynamic viscoelastometer and profilometer, respectively. In addition, weight changes during immersion in water were determined. RESULTS: The working time and gelation time of the alcohol-free tissue conditioner were similar to those of the conventional materials. This alcohol-free material had significantly lower shear storage modulus and shear loss modulus, and higher loss tangent (P<0.05) than FITT and SR-Ivoseal at 0.01 and 1 Hz. The alcohol-free material maintained its inherent viscoelastic properties and exhibited only a slight change in weight during 14 days of water immersion when compared to the conventional materials. The application of the coating significantly reduced the loss of the initial viscoelastic properties and surface quality during the test periods. CONCLUSIONS: The coated alcohol-free tissue conditioner would be superior to the conventional materials containing ethyl alcohol in view of viscoelastic properties after gelation, compatibility with dental stones and durability. PMID- 16216405 TI - Antiangiogenic and proapoptotic activity of a novel glycoprotein from U. indica is mediated by NF-kappaB and Caspase activated DNase in ascites tumor model. AB - We have identified a novel glycoprotein from Urginea indica bulbs with potent in vivo antitumor activity against growth of an ascites tumor, mouse mammary carcinoma. In this paper we report characterization of a 29 kDa glycoprotein from U. indica and demonstrate the mechanism of antiangiogenic and proapoptotic activity. N-terminal sequence of the high performance liquid chromatography (HPLC) pure glycoprotein showed sequence homology to an extent of 40-50% with known angiogenesis inhibitor and apoptosis-inducing protein from C. elegans and G. gallus respectively. Our results on antiangiogenic property of the glycoprotein include inhibition of in vivo angiogenesis assays, decreased micro vessel density count and CD31 antigen staining in 29 kDa glycoprotein treated mice peritoneum. In vitro inhibition of vascular endothelial growth factor induced proliferation of human umbilical vein endothelial cells (HUVECs) by the glycoprotein further supports its antiangiogenic activity. The mechanism of antiangiogenesis involved inhibition of translocation of nuclear factor kappa B to the nucleus resulting in decreased expression of vascular endothelial growth factor gene as is demonstrated by our results on quantification of vascular endothelial growth factor levels in the glycoprotein treated tumor bearing mice. Our results on activation of Caspase-3 with concomitant translocation of caspase activated DNase to the tumor cell nuclei resulting in DNA fragmentation induced by the glycoprotein in vivo clearly demonstrated a parallel proapoptotic activity of the glycoprotein. PMID- 16216406 TI - Failure of normal glycaemic regulation in a patient with severe hypothermia. AB - This report describes the case of an 88-year-old non-diabetic female who presented to the emergency department following a presumed hypoglycaemic collapse due to self-neglect. Subsequent rewarming and resuscitation demonstrated a number of the significant consequences of severe hypothermia, including apparent secondary impairment of glycaemic autoregulation. The phenomenon of reversible inhibition of insulin secretion due to severe hypothermia has been documented previously in the field of cardiac surgery. The hyperglycaemia was not treated with any antihyperglycaemic agent, and her recovery was uneventful. Subsequent blood sugar level monitoring was normal. If insulin is administered to the hypothermic patient, intensive monitoring of blood glucose is essential due to the increase in endogenous insulin secretion on rewarming. PMID- 16216407 TI - Effects of switching between leftward and rightward pro- and antisaccades. AB - Previous studies suggested that random switching between pro- and antisaccades increases errors in both tasks. However, little is known about the effects of switching between leftward and rightward saccades (response switching). The present study investigated task and response switching using an alternating runs procedure. Tasks (i.e., prosaccades versus antisaccades) were switched every second trial. Response switches (i.e., leftward saccades versus rightward saccades) were counterbalanced across tasks and task-switching conditions. Task switching increased errors in both tasks. Response switching increased errors when antisaccades were preceded by antisaccades but not when antisaccades were preceded by prosaccades or for prosaccades regardless of the preceding saccade type. The task-switch effects suggest that both pro- and antisaccade trials activate specific production rules that can persist in a subsequent trial. The differential response-switch effects may reflect different modes of response activation in pro- and antisaccades (sensorimotor transformation of visual information versus selection of motor programs). PMID- 16216408 TI - The effect of a nicorandil congener on isolated human myometrium. AB - OBJECTIVE: To determine the effect of a nicorandil congener, SG-209 on the myometrium. STUDY DESIGN: Isolated strips of myometrium, from nine women who underwent hysterectomy, were made to contract with 55 mM KCl before and after incubation with three concentrations of SG-209 (1.5, 3.0 and 10 microM). The mean height of contraction and the area under the curve were analysed using a non parametric test. RESULTS: At a 10 microM concentration, SG-209 significantly decreased the mean area under the curve from 10.8 to 2.5 cm2 (p<0.05). CONCLUSION: SG-209 significantly relaxes the human non-pregnant myometrium. Along with its congener, nicorandil, it may be useful in clinical conditions that require uterine relaxation. PMID- 16216410 TI - Increased protein expression of both inducible nitric oxide synthase and cyclooxygenase-2 in human colon cancers. AB - The present study was conducted to investigate protein levels of inducible NO synthase (iNOS) and cayclooxygenase-2 (COX-2) in human colon cancer tissues. Significant increase of iNOS and COX-2 levels in tumors as compared to paired normal tissue was observed in 15 and 28, respectively, of 29 cases investigated. No correlation was observed between the two enzymes, and immunohistochemically demonstrated iNOS expression was mainly observed in tumor epithelial cells and was higher in well-differentiated than moderately-differentiated adenocarcinomas, while COX-2 expression was mainly detected in stromal cells and higher in Dukes' C and D than Dukes' A and B cases. These results suggest that iNOS and COX-2 have separate roles in colon carcinogenesis, and might both be usefully targeted by chemoprevention. PMID- 16216409 TI - Overexpression of PML induced apoptosis in bladder cancer cell by caspase dependent pathway. AB - The promyelocytic leukemia gene (PML) encodes a growth/tumor suppressor protein that is essential for the induction of apoptosis in response to various apoptotic signals. The mechanism by which PML plays a role in the regulation of cell death is still unknown. Our previous study demonstrated that overexpression of PML suppress the growth of bladder cancer cells by inducing apoptosis and cell cycle arrest. To further elucidate the mechanism of PML induced apoptosis in bladder cancer, we constructed a PML inducible stable cell line. We found that the increased expression of PML significantly inhibit the growth of the UM-UC-2/PML clone cells and present apparent massive apoptosis in 24 h post-induction, while the UM-UC-2/PMEP4 cells are not. We also examined the effect of PML on the cell cycle distribution in UM-UC-2 cells. We showed overexpression of PML cause a cell cycle arrest in G1 phase. In additional, increased expression of PML in bladder cancer UM-UC-2 cells reduce Survivin expression and up regulated Caspase-3, and cleaved PARP expression, these suggested that PML might regulate apoptosis through Caspase dependent pathways. Our results demonstrate a novel mechanism of PML-induced apoptosis by down-regulation of Survivin and activation of Caspase dependent pathway. PMID- 16216411 TI - Molecular pathways in malignant pleural mesothelioma. AB - Malignant pleural mesothelioma, although uncommon, is highly lethal. There is a high correlation between associated environmental exposure factors, carcinogens, and its development. Carcinogenesis is also mediated by genetic defects that result in loss of tumor suppressors or over expression of proto-oncogenes. Factors such as the loss of CDK inhibition function, IGF stimulatory pathways, p14(ARF), p15(INK4b), p16(INK4a), p21, and p53 loss or mutation, VEGF and COX over expression are discussed. Correlations to potential therapeutic modalities are made. PMID- 16216412 TI - Photochemical treatment of phenol aqueous solutions using ultraviolet radiation and hydrogen peroxide. AB - The potential of purifying phenol aqueous solutions (0.0006-0.0064 M) using ultraviolet (UV) radiation and hydrogen peroxide (H2O2; 0.005-0.073 M) was investigated. Although the direct photolysis of phenol and its oxidation by hydrogen peroxide (without ultraviolet light) were insignificant, the combination of UV and H2O2 was extremely effective on phenol degradation. However, the chemical oxygen demand was on no occasion entirely eliminated, indicating the resistance of the intermediate products formed to the photo-oxidation. Increasing the initial concentration of phenol had as a result lower phenol conversions achieved, whereas the increase in hydrogen peroxide initial concentration enhanced significantly the degradation of phenol. In contrast, COD removal was less sensitive to these changes. PMID- 16216413 TI - Functional role of JNK in neuritogenesis of PC12-N1 cells. AB - JNKs, also known as SAPKs, are activated in response to a wide variety of factors including growth factors, cytokines, UV radiation, and heat shock. In the rat pheochromocytoma PC12 cells, the JNK signaling pathway mediates diverse functions such as differentiation and apoptosis. We have previously shown that activated JNK is required for later stages of neuritogeneis induced by NGF in a variant PC12 cell line (N1). Here, the functional role of JNK in N1 cells is further investigated. We show that NGF treatment, which induces extensive neurite branching and cell soma enlargement in the N1 cells, stimulates a biphasic activation of JNK. The first phase of activation is rapid and transient, beginning at 15 min after NGF exposure and lasting approximately 45 min. The second phase of activation is sustained, beginning at 9-12 h of NGF treatment and lasting for at least 24 h. Similar biphasic pattern of JNK activation is also observed in the parental PC12 cells. Using the specific JNK inhibitor SP600125, we show that the biphasic activation is necessary for neurite outgrowth and branching, and that inhibition of either phase suppresses neuritogenesis in the N1 cells. These results suggest that dynamic JNK activation may play a key role in neurite outgrowth during neuronal development. PMID- 16216414 TI - Involvement of neuropeptide Y and Y1 receptor in antinociception in the arcuate nucleus of hypothalamus, an immunohistochemical and pharmacological study in intact rats and rats with inflammation. AB - Neuropeptide Y (NPY) plays an important role in pain modulation at different levels in the central nervous system. In the brain, NPY and NPY receptors distribute abundantly in the arcuate nucleus of hypothalamus (ARC), a structure involved in pain processing. The present study was undertaken to investigate the role of NPY in nociceptive modulation in the ARC of intact rats and rats with carrageenan-induced inflammation. Intra-ARC administration of NPY induced dose dependent increases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation in intact rats, which was attenuated by the Y1 receptor antagonist NPY28-36. Intra-ARC administration of NPY also induced dose-dependent increases in HWLs to noxious stimulation in rats with inflammation. Furthermore, intra-ARC injection of either the antiserum against NPY or NPY28-36 induced decreases in HWLs in rats with inflammation, while both of them produced no effects in intact ones. Additionally, there were marked increases of Y1 receptor in the bilateral ARC of rats with inflammation tested by immunohistochemistry, while no significant changes of NPY were observed, implicating that the increased Y1 receptor has an important effect in the NPY-induced antinociception. We also found that intra-ARC injection of Y2 receptor agonist NPY3-36 produced no significant antinociception in either intact rats or rats with inflammation. Together, we demonstrate that NPY exerts an antinociceptive effect in the ARC of intact rats and rats with inflammation. Both Y1 receptor and endogenous released NPY in the ARC are involved in the nociceptive modulation during inflammation. PMID- 16216416 TI - Screening for and purification of novel self-aggregatable lectins reveal a new functional lectin group in the bark of leguminous trees. AB - A solubility-insolubility transition assay was used to screen the bark and stems of seven leguminous trees and plants for self-aggregatable lectins. Novel lectins were found in two trees, Robinia pseudoacacia and Wisteria floribunda, but not in the leguminous plants. The Robinia lectin was isolated from coexisting lectin by combined affinity chromatographies on various sugar adsorbents. The purified lectins proved to be differently glycosylated glycoproteins. One lectin exhibited the remarkable characteristics of self-aggregatable lectins: localization in the bark of legume trees, self-aggregation dissociated by N acetylglucosamine/mannose, and coexistence with N-acetylgalactosamine/galactose specific lectins, which are potential endogenous receptors. Self-aggregatable lectins are a functional lectin group that can link enhanced photosynthesis to dissociation of glycoproteins. PMID- 16216417 TI - A comparative study of neuronal and inducible nitric oxide synthases: generation of nitric oxide, superoxide, and hydrogen peroxide. AB - Nitric oxide synthases (NOS) independent of the isozyme, produce nitric oxide (.NO), superoxide (O2.-), and hydrogen peroxide (H2O2). Since .NO has been implicated in many physiological processes, the importance of O2.- and H2O2 in regulating cell signaling by .NO cannot be overlooked. Before addressing these questions, we investigated the production of .NO, O2.-, and H2O2 by purified NOS. NOS 1 and NOS 2 were chosen, as the flux of .NO from each isozyme supports differential biological activity. We found that the initial rate and sustained production of .NO was considerably greater for NOS 2 as compared to NOS 1. In the absence of L-arginine, however, NOS 1 generation of O2.- and H2O2 was found to be substantially greater than that measured for NOS 2. Differences between NOS 1 and NOS 2 production of .NO, O2.-, and H2O2 may define the specific physiologic function of each isozyme. PMID- 16216419 TI - Hepatocellular carcinoma as a complex polygenic disease. Interpretive analysis of recent developments on genetic predisposition. AB - The different frequency of hepatocellular carcinoma (HCC) in humans at risk suggests a polygenic predisposition. However, detection of genetic variants is difficult in genetically heterogeneous human population. Studies on mouse and rat models identified 7 hepatocarcinogenesis susceptibility (Hcs) and 2 resistance (Hcr) loci in mice, and 7 Hcs and 9 Hcr loci in rats, controlling multiplicity and size of neoplastic liver lesions. Six liver neoplastic nodule remodeling (Lnnr) loci control number and volume of re-differentiating lesions in rat. A Hcs locus, with high phenotypic effects, and various epistatic gene-gene interactions were identified in rats, suggesting a genetic model of predisposition to hepatocarcinogenesis with different subset of low-penetrance genes, at play in different subsets of population, and a major locus. This model is in keeping with human HCC epidemiology. Several putative modifier genes in rodents, deregulated in HCC, are located in chromosomal segments syntenic to sites of chromosomal aberrations in humans, suggesting possible location of predisposing loci. Resistance to HCC is associated with lower genomic instability and downregulation of cell cycle key genes in preneoplastic and neoplastic lesions. p16(INK4A) upregulation occurs in susceptible and resistant rat lesions. p16(INK4A)-induced growth restraint was circumvented by Hsp90/Cdc37 chaperons and E2f4 nuclear export by Crm1 in susceptible, but not in resistant rats and human HCCs with better prognosis. Thus, protective mechanisms seem to be modulated by HCC modifiers, and differences in their efficiency influence the susceptibility to hepatocarcinogenesis and probably the prognosis of human HCC. PMID- 16216418 TI - Glutathione turnover in human cell lines in the presence of agents with glutathione influencing potential with and without acivicin inhibition of gamma glutamyltranspeptidase. AB - BACKGROUND: We have previously shown that there were great discrepancies between different agents regarding their glutathione stimulating potential and that agents with mainly oxidative effects did not increase concentrations of glutathione in human cell cultures, in contrast to other thiol reactive agents. In order to evaluate whether increased glutathione degradation might be one reason for these discrepancies, we have investigated the effect of different agents with potential influence on glutathione metabolism in human cell cultures with or without acivicin inhibition of gamma-glutamyltranspeptidase (GT), since GT is responsible for the initial degradation of glutathione. METHODS: Intra- and extracellular concentrations of glutathione were investigated in HeLa and hepatoma cell cultures, with and without acivicin inhibition of GT, in the presence of oxidative and electrophilic agents (copper ions, hydrogen peroxide and N-ethylmaleimide), hydroquinone, reducing agents (lipoic acid and N acetylcysteine), and a thiol reactive metal (mercury ions). RESULTS: There were great discrepancies between the different agents regarding their maximal glutathione response (the sum of the intracellular and the extracellular amount of glutathione) in cell cultures. There was only a small increase in total glutathione in the presence of hydrogen peroxide or N-ethylmaleimide before the cell protein decreased compared to findings with mercury ions, lipoic acid or hydroquinone. In both HeLa and hepatoma cell cultures, there were correlations between the original glutathione amount and the total glutathione amount observed after acivicin inhibition. CONCLUSION: The relatively small increase of glutathione amount in the presence of oxidative and electrophilic agents compared to other thiol reactive agents is not due to increased GT degradation of glutathione. PMID- 16216420 TI - Vesicular glutamate transporters type 1 and 2 expression in axon terminals of the rat nucleus of the solitary tract. AB - The nucleus of the solitary tract is the site of termination of primary afferent fibers running in the facial, glossopharyngeal and vagus nerves. The present study was performed to map the distribution of glutamatergic axons terminals in the rat nucleus of the solitary tract using immunodetection of vesicular glutamate transporter 1 and vesicular glutamate transporter 2. The two vesicular glutamate transporters were differentially distributed among nucleus of the solitary tract subdivisions. Vesicular glutamate transporter 1 immunoreactivity was mostly found in the lateral part of the nucleus (ventrolateral, interstitial and intermediate subdivisions) whereas vesicular glutamate transporter 2 labeling was distributed throughout the nucleus of the solitary tract. Electron microscope examination indicated that vesicular glutamate transporter immunoreactivity was localized in axon terminals filled with round synaptic vesicles. After injection of cholera toxin B subunit in sensory ganglia, anterograde labeling was found in vesicular glutamate transporter 1, as well as vesicular glutamate transporter 2 immunoreactive boutons. Double immunolabeling experiments allowed distinctions between terminals expressing either vesicular glutamate transporter 1 or vesicular glutamate transporter 2 or both vesicular glutamate transporter 1 and vesicular glutamate transporter 2 immunoreactivities. The latter population, expressing both transporters immunolabeling, completely disappeared after deafferentation induced by removal of sensory ganglia. This study indicates that vesicular glutamate transporter content identifies three different subpopulations of glutamatergic boutons in the nucleus of the solitary tract and provides definitive evidence that primary afferent neurons contribute glutamatergic terminals to the nucleus of the solitary tract. PMID- 16216421 TI - A study of the role of neuro-glial remodeling in the oxytocin system at lactation. AB - Under conditions of strong secretion of neurohypophysial hormone, such as during parturition, lactation and dehydration, the hypothalamic oxytocin-system displays a remarkable morphological plasticity such that astrocytic coverage of its neurones diminishes, their surfaces become directly juxtaposed and contacted by an increased number of synapses. A growing body of evidence indicates that these anatomical changes have an impact on glutamatergic neurotransmission in the supraoptic nucleus, and may be therefore of physiological consequence. We here evaluated the consequences of the inhibition of such plasticity on the overall activity of the oxytocin system during lactation. Remodeling was prevented by performing hypothalamic microinjections in gestating rats of endoneuraminidase, an enzyme that removes polysialic acid from the neural cell adhesion molecule. Our earlier studies established that the presence of polysialic acid is a prerequisite for remodeling of the oxytocin system in the supraoptic and paraventricular nuclei. In dams in which polysialic acid was absent in all magnocellular nuclei after bilateral endoneuraminidase injections, parturition was normal and neither the frequency nor the amplitude of suckling-induced reflex milk ejections was different from vehicle-treated dams. The weight gain of pups was also normal as was water intake by the dams. We then assessed the electrical activity of antidromically identified magnocellular neurones in the polysialic acid-free supraoptic nucleus of isoflurane-anesthetized lactating rats. Basal and bursting activity characteristic of oxytocin neurones before each reflex milk ejection was not significantly different from that recorded in the supraoptic nucleus of rats with normal levels of polysialic acid. Our results indicate that neuro-glial remodeling, despite its role on fine modulation of oxytocin neuronal activity, is not essential to parturition and lactation. PMID- 16216422 TI - Spectral integration in the inferior colliculus of the CBA/CaJ mouse. AB - The inferior colliculus receives a massive convergence of inputs and in the mustached bat, this convergence leads to the creation of neurons in the inferior colliculus that integrate information across multiple frequency bands. These neurons are tuned to multiple frequency bands or are combination-sensitive; responding best to the combination of two signals of different frequency composition. The importance of combination-sensitive neurons in processing echolocation signals is well described, and it has been thought that combination sensitivity is a neural specialization for echolocation behaviors. Combination sensitivity and other response properties indicative of spectral integration have not been thoroughly examined in the inferior colliculus of non-echolocating mammals. In this study we tested the hypothesis that integration across frequencies occurs in the inferior colliculus of mice. We tested excitatory frequency response areas in the inferior colliculus of unanesthetized mice by varying the frequency of a single tone between 6 and 100 kHz. We then tested combination-sensitive responses by holding one tone at the unit's best frequency, and varying the frequency and intensity of a second tone. Thirty-two percent of the neurons were tuned to multiple frequency bands, 16% showed combination sensitive facilitation and another 12% showed combination-sensitive inhibition. These findings suggests that the neural mechanisms underlying processing of complex sounds in the inferior colliculus share some common features among mammals as different as the bat and the mouse. PMID- 16216424 TI - Acute phase gene expression in mice exposed to the marine neurotoxin domoic acid. AB - Domoic acid is a rigid analog of the neurotransmitter glutamate and a potent agonist of kainate subtype glutamate receptors. Persistent activation of these receptor subtypes results in rapid excitotoxicity, calcium dependent cell death and neuronal lesions in areas of the brain where kainate pathways are concentrated. To better understand responses to domoic acid induced excitotoxicity, microarrays were used to profile gene expression in mouse brain following domoic acid exposure. Adult female mice were subjected intraperitoneally to domoic acid at the lethal dose 50, killed and dissected at 30, 60 and 240 min post-injection. Total brain RNA from treated mice was compared with time-matched controls on Agilent 22K feature microarrays. Real-time PCR was performed on selected genes. For the 30, 60 and 240 min time points, 3.96%, 3.94% and 4.36% of the genes interrogated were differentially expressed (P-value < or = 0.01), respectively. Rigorous filtering of the data resulted in a set of 56 genes used for trending analysis and K-medians and agglomerative clustering. The earliest genes induced consisted primarily of early response gene families (Jun, Fos, Ier, Egr, growth arrest and DNA damage 45) and the inflammatory response element cyclooxygenase 2. Some later responding genes involved glucocorticoid responses (Gilz, Sgk), cold inducible proteins (Cirbp, Rbm3), Map kinases (Map3k6) and NF-kappaB inhibition. Real-time PCR in male mice from an additional study confirmed the expression of several of these genes across gender. The transcriptional profile induced by domoic acid shared similarity with expression profiles of brain ischemia and other excitotoxins, suggesting a common transcriptional response. PMID- 16216423 TI - Chronic exposure to typical or atypical antipsychotics in rodents: temporal effects on central alpha7 nicotinic acetylcholine receptors. AB - A decrease in alpha7 nicotinic acetylcholine receptors in the hippocampus has been hypothesized to contribute to alterations in auditory gating and other behavioral impairments in schizophrenia. However, while both typical and atypical neuroleptics are routinely used in the therapeutics of schizophrenia, little is known about their effects on auditory gating or alpha7 nicotinic acetylcholine receptor expression particularly when they are administered for extended periods of time (which is common in the clinical setting). In the present study in normal rats, the residual effects of prior chronic treatment (90 or 180 days) with representative typical and atypical neuroleptics (oral haloperidol, 2.0 mg/kg/day; chlorpromazine, 10.0 mg/kg/day, risperidone, 2.5 mg/kg/day; or olanzapine, 10.0 mg/kg/day) on prepulse inhibition of the auditory gating response were investigated. The densities of alpha7 nicotinic acetylcholine receptors were subsequently measured using [125I]-alpha-bungarotoxin autoradiography. The results indicated that none of the compounds significantly altered the startle amplitude or prepulse inhibition response either during drug treatment (day 60) or after 90 or 180 days of treatment (i.e. during a drug free washout). However, prior exposure to chlorpromazine, risperidone and olanzapine for 90 days resulted in modest but significant (P<0.01) decreases in [125I]-alpha bungarotoxin binding sites in some brain regions (e.g. posterior cortical amygdala). After 180 days of treatment, decreases in [(125I]-alpha-bungarotoxin binding ranging from approximately 12% (lateral dentate gyrus) up to 24% (e.g. CA1 hippocampal region) were evident in the risperidone group in 13 of the 36 regions analyzed while decreases associated with the other neuroleptics agents were still present, but not statistically significant. These data indicate that the commonly used atypical neuroleptic, risperidone is associated with time dependent and persistent negative effects on an important biological substrate of memory (i.e. the alpha7 nicotinic receptor), but that the magnitude of the deficits was not sufficient to impair auditory gating. PMID- 16216426 TI - Acute effects of ethanol on hippocampal long-term potentiation and long-term depression are mediated by different mechanisms. AB - To determine potential mechanisms contributing to ethanol-induced cognitive impairment, we examined acute effects of ethanol on hippocampal N-methyl-d aspartate receptors and forms of synaptic plasticity thought to underlie memory processing. In the CA1 region of rat hippocampal slices, ethanol partially inhibited N-methyl-d-aspartate receptor-mediated synaptic responses at concentrations up to 180 mM. The block of synaptic N-methyl-d-aspartate receptors by 60mM ethanol occluded the effects of 10 microM ifenprodil, an agent that has relative selectivity for N-methyl-D-aspartate receptors expressing NR1 and NR2B subunits. Ethanol did not occlude the effects of a low concentration of 2-amino-5 phosphonovalerate, an antagonist with less N-methyl-d-aspartate receptor subtype selectivity. Recent studies indicate that ifenprodil and other NR2B-selective antagonists inhibit N-methyl-D-aspartate receptor-dependent long-term depression but not long-term potentiation. We found that ethanol reversibly inhibited long term depression in a manner consistent with its effects on synaptic N-methyl-D aspartate receptors. Ethanol also inhibited the induction of N-methyl-D-aspartate receptor-dependent long-term potentiation, but the actions on long-term potentiation were complex and largely irreversible over the time course of our experiments. Furthermore, ethanol inhibited a form of long-term potentiation induced by very high frequency stimulation that does not depend on N-methyl-D aspartate receptor activation. The effects of ethanol on both forms of long-term potentiation, but not on long-term depression, were at least partially reversed by block of GABA type A receptors with picrotoxin. These results indicate that pharmacologically relevant concentrations of ethanol exert preferential effects on a subtype of synaptic N-methyl-D-aspartate receptors in the CA1 hippocampal region. Inhibition of synaptic N-methyl-D-aspartate receptors appears to contribute strongly to ethanol-mediated long-term depression inhibition, but effects on long-term potentiation are complex, involving, at least partially, changes in GABAergic transmission. PMID- 16216425 TI - A possible role for dopamine D3 receptor stimulation in the induction of neurogenesis in the adult rat substantia nigra. AB - Small molecule neurotransmitters, such as dopamine, have been shown to regulate cell cycles in the developing brain [Spencer GE, Klumperman J, Syed NI (1998) Neurotransmitters and neurodevelopment: Role of dopamine in neurite outgrowth, target selection and specific synapse formation. Perspect Dev Neurobiol 5:451 467; Ohtani N, Goto T, Waeber C, Bhide PG (2003) Dopamine modulates cell cycle in the lateral ganglionic eminence. J Neurosci 23:2840-2850] and may provide an alternative to traditional growth factors for the regulation of neurogenesis. Specifically, the dopamine D3 receptor appears to play an important role in neural development, and shows a persistent expression through adulthood in the proliferative subventricular zone [Diaz J, Ridray S, Mignon V, Griffon N, Schwartz JC, Sokoloff P (1997) Selective expression of dopamine D3 receptor mRNA in proliferative zones during embryonic development of the rat brain. J Neurosci 17:4282-4292]. Furthermore, pharmacological stimulation of D3 receptors promotes proliferation of adult subventricular zone cells, both in vitro [Coronas V, Bantubungi K, Fombonne J, Krantic S, Schiffmann SN, Roger M (2004) Dopamine D3 receptor stimulation promotes the proliferation of cells derived from the post natal subventricular zone. J Neurochem 91:1292-1301] and in vivo [Van Kampen JM, Hagg T, Robertson HA (2004) Induction of neurogenesis in the adult rat subventricular zone and neostriatum following dopamine D3 receptor stimulation. Eur J Neurosci 19:2377-2387]. In earlier work, we have demonstrated the induction of cell proliferation in the subventricular zone of the adult rat brain accompanied by a dramatic 10-fold induction of neurogenesis in the neighboring neostriatum, following administration of the preferential D3 receptor agonist, 7 hydroxy-N,N-di-n-propyl-2-aminotetralin [Van Kampen JM, Hagg T, Robertson HA (2004) Induction of neurogenesis in the adult rat subventricular zone and neostriatum following dopamine D3 receptor stimulation. Eur J Neurosci 19:2377 2387]. Dopamine D3 receptors have also been found in the substantia nigra [Diaz J, Pilon C, Le Foll B, Gross C, Triller A, Schwartz JC, Sokoloff P (2000) Dopamine D3 receptors expressed by all mesencephalic dopamine neurons. J Neurosci 20:8677-8684], a region of the adult brain shown to exhibit ongoing cytogenesis and neurogenic potential [Lie DC, Dziewczapolski G, Willhoite AR, Kaspar BK, Shults CW, Gage FH (2002) The adult substantia nigra contains progenitor cells with neurogenic potential. J Neurosci 22:6639-6649; Zhao M, Momma S, Delfani K, Carlen M, Cassidy RM, Johansson CB, Brismar H, Shupliakov O, Frisen J, Janson AM (2003) Evidence for neurogenesis in the adult mammalian substantia nigra. Proc Natl Acad Sci U S A 100:7925-7930]. We have found that chronic intraventricular administration of 7-hydroxy-N,N-di-n-propyl-2-aminotetralin triggers a profound induction of cell proliferation in the rat substantia nigra and promotes the adoption of a neuronal phenotype in a proportion of these newly generated cells. PMID- 16216427 TI - Increase in proliferation and gliogenesis but decrease of early neurogenesis in the rat forebrain shortly after transient global ischemia. AB - Regarding regenerative strategies early post-ischemic therapeutic interventions might have a great impact on further pathophysiological cascades. To understand the early post-ischemic events we analyzed proliferation and neurogenesis as early as on day 3 after transient global ischemia in rats. Evaluations were performed not only in the dorsal hippocampus, where post-ischemic cell death develops selectively in the cornu ammonis, subfield 1 area, but also in distant areas like the ventricle wall and the striatum. Ischemia was induced by a transient two-vessel occlusion combined with hypotension. Animals received daily i.p. injections of 5-bromo-2-deoxyuridine until decapitation 1 or 3 days after ischemia. Immunohistochemistry was performed to detect 5-bromo-2-deoxyuridine and co-labeling with cell-specific markers. Three days after ischemia, proliferation significantly increased throughout the forebrain. Early neurogenesis, detected by doublecortin labeling, on the other hand, was restricted to the neurogenic zones of the dentate gyrus and the lateral ventricle. Global ischemia reduced the overall number of doublecortin-positive cells in the dentate gyrus, particularly in the upper blade of the dentate gyrus. However, the number of newly generated doublecortin- and 5-bromo-2-deoxyuridine double-labeled cells was unchanged. The vast majority of newly generated cells were microglia/macrophages, which invaded morphologically damaged as well as undamaged regions. Astroglial cells were activated all over the forebrain by the ischemic insult. They were co-localized almost completely with nestin in many areas, yet, sparsely proliferated after the insult. Interestingly, in locally defined zones we found nestin- and glial fibrillary acidic protein-signals clearly separated. In sham-operated animals, nestin could be detected in both neurogenic zones only without co-labeling with glial markers. In conclusion, during the first days after global ischemia, cell death of cornu ammonis, subfield 1-neurons was accompanied by a massive overall proliferation and activation of microglia/macrophages, a reduction of pre ischemia existing doublecortin-positive precursors in the dentate gyrus and a re expression of nestin in glial fibrillary acidic protein-positive astrocytes. PMID- 16216429 TI - Expression of neuronal and inducible nitric oxide synthase in neuronal and glial cells after transient occlusion of the middle cerebral artery. AB - We presently investigated the time-course of neuronal nitric oxide synthase and inducible nitric oxide synthase expression and content in the rat striatum up to 6 days after ischemia induced by transient middle cerebral artery occlusion, a condition that potentially allows functional recovery, with the aim to identify the cell types expressing these two enzymes and to correlate neuronal nitric oxide synthase and inducible nitric oxide synthase changes in order to verify whether and how these changes are related to tissue damage, motor-sensory performances and survival. Before and after surgery, the animals underwent neurological evaluation. The results demonstrated that the rats with a score > or = 12 at the neurological evaluation 24 h after ischemia showed a significant increase in neuronal nitric oxide synthase-immunoreactive neurones and absence of inducible nitric oxide synthase-immunoreactive cells and survived up to the sixth day; conversely, the rats with a score < 12 at the neurological evaluation 24 h after ischemia showed a progressive significant decrease in neuronal nitric oxide synthase-immunoreactive neurones and appearance of inducible nitric oxide synthase-immunoreactive cells and none of the rats survived up to the sixth day. Microglia cells were activated in both groups but only in the latter did these cells express inducible nitric oxide synthase. Measurement of the infarct area demonstrated that it occupied a similar territory in both groups of rats but in those with a score < 12 the edema was more extended. In conclusion, we demonstrated that a neurotoxic insult such as ischemia can induce neuronal nitric oxide synthase expression in the neurones and that when neuronal nitric oxide synthase-immunoreactive neurones increase in number, microglia activation is less extended, inducible nitric oxide synthase-immunoreactive cells are absent, tissue damage reduced and the rats survive longer. Conversely, when there is a significant decrease of neuronal nitric oxide synthase-immunoreactive neurones, microglia cells are intensely activated, inducible nitric oxide synthase immunoreactive cells appear and the animal survival is shortened. PMID- 16216428 TI - Endogenous neuropeptide Y depresses the afferent signaling of gastric acid challenge to the mouse brainstem via neuropeptide Y type Y2 and Y4 receptors. AB - Vagal afferents signal gastric acid challenge to the nucleus tractus solitarii of the rat brainstem. This study investigated whether nucleus tractus solitarii neurons in the mouse also respond to gastric acid challenge and whether this chemonociceptive input is modified by neuropeptide Y acting via neuropeptide Y receptors of type Y2 or Y4. The gastric mucosa of female mice was exposed to different concentrations of HCl or saline, excitation of neurons in the nucleus tractus solitarii visualized by c-Fos immunohistochemistry, gastric emptying deduced from the gastric volume recovery, and gastric lesion formation evaluated by planimetry. Relative to saline, intragastric HCl (0.15-0.35 M) increased the number of c-Fos-expressing cells in the nucleus tractus solitarii in a concentration-dependent manner, inhibited gastric emptying but failed to cause significant hemorrhagic injury in the stomach. Mice in which the Y2 or Y4 receptor gene had been deleted responded to gastric acid challenge with a significantly higher expression of c-Fos in the nucleus tractus solitarii, the increases amounting to 39 and 31%, respectively. The HCl-induced inhibition of gastric emptying was not altered by deletion of the Y2 or Y4 receptor gene. BIIE0246 ((S)-N2-[[1-[2-[4-[(R,S)-5,11-dihydro-6(6H)-oxodibenz[b,e] azepin-11-yl] 1-piperazinyl]-2-oxoethyl]cyclopentyl] acetyl]-N-[2-[1,2-dihydro-3,5 (4H)-dioxo 1,2-diphenyl-3H-1,2,4-triazol-4-yl]ethyl]-argininamide; 0.03 mmol/kg s.c.), a Y2 receptor antagonist which does not cross the blood-brain barrier, did not modify the c-Fos response to gastric acid challenge. The Y2 receptor agonist peptide YY (3-36) (0.1 mg/kg intraperitoneally) likewise failed to alter the gastric HCl evoked expression of c-Fos in the nucleus tractus solitarii. BIIE0246, however, prevented the effect of peptide YY-(3-36) to inhibit gastric acid secretion as deduced from measurement of intragastric pH. The current data indicate that gastric challenge with acid concentrations that do not induce overt injury but inhibit gastric emptying is signaled to the mouse nucleus tractus solitarii. Endogenous neuropeptide Y acting via Y2 and Y4 receptors depresses the afferent input to the nucleus tractus solitarii by a presumably central site of action. PMID- 16216430 TI - Midbrain muscarinic receptors modulate morphine-induced accumbal and striatal dopamine efflux in the rat. AB - Midbrain dopamine neurons are critical in mediating the rewarding effects of opiates in dependent rats, as well as modulating some manifestations of opiate withdrawal. Morphine is known to excite dopamine neurons and thereby facilitate forebrain dopamine transmission through inhibition of GABA neurons. Cholinergic neurons in the mesopontine laterodorsal and pedunculopontine tegmental nuclei provide the principal source of excitatory cholinergic input to ventral tegmental area and substantia nigra pars compacta dopamine-containing neurons, via actions on midbrain muscarinic and nicotinic acetylcholine receptors. The present study hypothesized that a reduction in tonic cholinergic input via blockade of midbrain muscarinic receptors would reduce the pharmacological effects of morphine on forebrain dopamine release. Using in vivo chronoamperometry, alterations in morphine-evoked dopamine efflux were monitored at stearate-graphite paste electrodes implanted unilaterally in the nucleus accumbens and striatum of urethane (1.5 g/kg) anesthetized rats, following the pharmacological inhibition of ventral tegmental area/substantia nigra pars compacta muscarinic receptors. The facilitatory effects of morphine (2.0 mg/kg, i.v.) on accumbens and striatal dopamine efflux were markedly reduced by prior infusion of the non-selective muscarinic receptor antagonist scopolamine (200 microg/microl) into the ventral tegmental area or substantia nigra pars compacta, respectively. These findings demonstrate that decreased activation of midbrain muscarinic receptors attenuates the excitatory effects of morphine on mesoaccumbens and nigrostriatal dopaminergic transmission. PMID- 16216431 TI - PSA-NCAM expression in the rat medial prefrontal cortex. AB - The rat medial prefrontal cortex, an area considered homologous to the human prefrontal cortex, is a region in which neuronal structural plasticity has been described during adulthood. Some plastic processes such as neurite outgrowth and synaptogenesis are known to be regulated by the polysialylated form of the neural cell adhesion molecule (PSA-NCAM). Since PSA-NCAM is present in regions of the adult CNS which are undergoing structural remodeling, such as the hypothalamus or the hippocampus, we have analyzed the expression of this molecule in the medial prefrontal cortex of adult rats using immunohistochemistry. PSA-NCAM immunoreactivity was found both in cell bodies and in the neuropil of the three divisions of the medial prefrontal cortex. All cell somata expressing PSA-NCAM corresponded to neurons and 5' bromodeoxyuridine labeling after long survival times demonstrated that these neurons were not recently generated. Many of these PSA-NCAM immunoreactive neurons in the medial prefrontal cortex could be classified as interneurons on the basis of their morphology and glutamate decarboxylase, isoform 67 expression. Some of the PSA-NCAM immunoreactive neurons also expressed somatostatin, neuropeptide Y and calbindin-D28K. By contrast, pyramidal neurons in this cortical region did not appear to express PSA-NCAM. However, some of these principal neurons appeared surrounded by PSA-NCAM immunoreactive puncta. Some of these puncta co-expressed synaptophysin, suggesting the presence of synapses. Since the etiology of some psychiatric disorders has been related to alterations in medial prefrontal cortex structural plasticity, the study of PSA-NCAM expression in this region may open a new approach to the pathophysiology of these mental disorders. PMID- 16216432 TI - Identification of gene markers based on well validated and subcategorized stressed animals for potential clinical applications in PTSD. AB - Post-traumatic stress disorder (PTSD) is a complex mental disorder that can develop in response to traumatic experiences. The molecular mechanisms underlying the pathology of PTSD are poorly understood, and this lack of knowledge hampers our ability to find superior therapeutic approaches to the treatment of this disorder. There are two main reasons for our lack of study in this area: here is no sufficiently validated animal model and lack of large-scale studies for the search of underlying molecular mechanisms. Thus, to promote research on PTSD (especially its molecular mechanisms) and to set molecular basis for searching novel medications of this disorder, large-scale, genome-wide interrogation of a significant amount of genes based upon a well validated animal model is demanded. We hypothesize that a significant number of genes are involved in PTSD. It is only with a large number of these genes identified in specific samples of PTSD related population, and then it is possible for a sufficient understanding of the pathology at the molecular level of a PTSD, as well as for enhancing the PTSD's therapeutic and preventative strategies. Two prerequisites are needed for testing this hypothesis: (1) relative pure samples from a well validated animal model; and (2) genome-wide screening of PTSD molecular targets. For the animal model, we suggest to use the predator-exposure paradigm, in which rats are exposed to a predator, this model has previously been evaluated behaviorally well emulated the clinical symptoms of PTSD. For a better stringency, three criteria can be used to further validate this animal model: analogous (similarity of behavior), predictive (predictability of drug response) and biological mechanism (e.g., electrophysiological and pathological change in amygdala). For large-scale molecular target screening, the new microarray technology, which can profile expression of tens of thousands genes simultaneously, is the method of choice. The validity and practicability of this hypothesis and the strategy for its testing have been supported by our preliminary laboratory data. PMID- 16216433 TI - Minodronate, a nitrogen-containing bisphosphonate, is a promising remedy for treating patients with diabetic retinopathy. AB - In diabetes mellitus, the formation and accumulation of advanced glycation end products (AGEs) progress. There is a growing body of evidence to show that AGEs their receptor (RAGE) interactions are involved in the development and progression of diabetic retinopathy. Bisphosphonates are potent inhibitors of bone resorption and are widely used drugs for the treatment of osteoporosis and osteolytic bone metastasis. Recently, farnesyl pyrophosphate synthase has been shown as a molecular target of nitrogen-containing bisphosphonates, and inhibition of post-translational prenylation of small molecular weight G proteins is likely involved in their anti-resorptive activity on osteoclasts. NADPH oxidase-derived reactive oxygen species (ROS) generation is required for the AGE RAGE signaling in vascular wall cells, and small G protein Rac is a critical component of vascular NADPH oxidase complex. These observations let us to speculate that minodronate, a newly developed nitrogen-containing bisphosphonate, might be a promising remedy for treating patients with diabetic retinopathy by inhibiting the AGE-RAGE signaling pathways through suppression of ROS generation via inhibition of Rac prenylation. In this paper, we like to propose the possible ways of testing our hypotheses: (1) Does treatment with minodronate decrease the risk for the development and progression of diabetic retinopathy in osteoporotic patients? (2) If the answer is yes, is this beneficial effect of minodronate superior to that of other nitrogen-noncontaining bisphosphonates with equihypolipidemic properties? (3) Does minodronate treatment suppress NADPH oxidase-mediated ROS generation in retinas of diabetic animals? (4) Does treatment with pyridoxamine, a post-Amadori inhibitor of AGE formation, attenuate these beneficial effects of minodronate on diabetic retinopathy? These clinical and animal studies could clarify whether the use of minodronate is of benefit in patients with AGE-RAGE-related disorders such as diabetic retinopathy, even in the absence of osteoporosis. PMID- 16216434 TI - Chloroquine-induced nitric oxide improves insulin sensitivity in rheumatoid arthritis. PMID- 16216435 TI - An hypothesis about genome structures in mammalian polyploid cells based on a new concept that genome is fractal of six hierarchies. AB - A new model for the spatial configurations of DNA is proposed to solve the problem of DNA loss in mammalian polyploid cells. The ordinary concept that chromosomes are situated independently in nuclei cannot account well for the DNA loss in polyploid cells. A new concept about the DNA configurations in diploid cells is constructed based on observations that have been reported. Briefly, the DNA structure is self-similar fractal with a unit of opposite-handed twin circles. In human diploid cells, DNA is constructed with six hierarchies whose sizes are 32(5), 32(4), 32(3), 32(2), 32(1) and 32(0) with a unit of 200 DNA base pairs, corresponding to a genome, a chromosome, a chromosome band, a replicon, a rosette loop (a gene) and a nucleosome, respectively. A model assuming particular spatial configurations of chromosomes in polyploid cells is deduced from this new concept about chromosome configurations in diploid cells. It can account satisfactorily for the problem of DNA loss in polyploid cells. When cell division is inhibited and DNA synthesis progresses, replicated DNA will be stacked. When inhibitors are removed, the polyploidized cells may return to the initial ploidy, because the stacked DNA loops have not been linked. When the stacked DNA twin loops are linked with a proper configuration, the cells may become polyploid cells. There is a distinct difference in genome structure between polyploidized and polyploid cells. The homologous chromosomes of polyploid cells are arrayed mirror-symmetrically and they can come close to each other in the folded structure. If DNA synthesis is bypassed at the paired homologous chromosomes, DNA content is lost at every cell division. As the DNA loss progresses, the chromosome configuration of polyploid cells deviates gradually from mirror symmetry and the DNA loss ceases, resulting in the establishment of semi-stable hypoploid. PMID- 16216436 TI - Atherosclerosis in developing and developed countries: relation with immune dysregulation and hygiene hypothesis. PMID- 16216437 TI - Clinical, phenotypic, and genotypic evidence for Streptococcus sinensis as the common ancestor of anginosus and mitis groups of streptococci. AB - In 2002, we reported the discovery of a novel species of viridans streptococcus, Streptococcus sinensis. Recently, we reported the isolation of two more strains of S. sinensis. Clinically, S. sinensis is a definite cause of infective endocarditis, a characteristic mainly pertaining to the mitis group of streptococci. Phenotypically, two of the three S. sinensis isolates were Lancefield group F, a characteristic of the anginosus group. However, none of the three strains possess the caramel smell typical of this group of streptococci. Biochemically, S. sinensis was identified in 56% of the time as members of the anginosus group, and in 33% of the time as members of the mitis group. These clinical and phenotypic properties should be governed by the presence/absence or expressivity of particular genes in the S. sinensis genome. Genotypically, phylogenetic analysis using 16S rRNA gene sequences showed that S. sinensis branched out as the first branch in the anginosus group, implying that it is the ancestor of the other members of this group. However, the bootstrap value for S. sinensis clustered with members of the anginosus group is only 47%, meaning that it is often not clustered with members of this group, but the mitis group. Furthermore, the differences in the 16S rRNA gene sequences between S. sinensis and Streptococcus intermedius (3.7%) and those between S. sinensis and Streptococcus gordonii (3.6%) are almost the same. All these indicated that it is very likely that S. sinensis is the common ancestor of the anginosus and mitis groups of streptococci. Complete genome sequencing of S. sinensis and comparative genomics studies on the S. sinensis genome and genomes of members in the anginosus and mitis groups should reveal clues to the underlying genotypic differences that govern the different phenotypic properties of the two groups of streptococci, such as why streptococci of the anginosus group are prone to cause abscess formation but not infective endocarditis as compared to other viridans streptococci. PMID- 16216438 TI - Nitric oxide and chronic fatigue syndrome: Are we caring for our patients or are we practicing selfcare? PMID- 16216439 TI - Hazardous alcohol consumption and sense of coherence in emergency department patients with minor trauma. AB - BACKGROUND: The concept of sense of coherence (SOC) measures the extent to which people are able to cope with stressful life events. Since excessive alcohol consumption may be, in part, the result of failed coping strategies, further investigations between hazardous alcohol consumption and SOC are necessary. METHODS: Survey in an Emergency Department in an inner city university hospital. Overall, 1,833 patients were screened for hazardous alcohol consumption, illicit drug use, smoking status, socioeconomic status and sense of coherence. RESULTS: Mean age of participants was 34+/-12 years and 62% were male. Overall, 23% of patients showed a pattern of hazardous alcohol consumption, 23% had abused illicit drugs and 46% were smokers. Mean SOC score was 48.9+/-8.6 points, with no difference between genders. Lower SOC-quartiles were associated with significantly higher prevalence of hazardous alcohol consumption as well as illicit drug abuse and smoking. Lower SOC-quartiles were associated with significantly lower values on socioeconomic variables (e.g. lower income, less education). The negative association between SOC and alcohol consumption was independent of age, gender, smoking, illicit drug use and all socioeconomic variables. CONCLUSION: SOC and hazardous alcohol consumption were significantly associated. Our data support Antonovsky's original concept of increased alcohol consumption as a result of failed coping strategies. PMID- 16216441 TI - Influence of the endosymbiont of Blastocrithidia culicis and Crithidia deanei on the glycoconjugate expression and on Aedes aegypti interaction. AB - Blastocrithidia culicis and Crithidia deanei are trypanosomatid protozoa of insects that normally contain intracellular symbiotic bacteria. The protozoa can be rid of their endosymbionts by antibiotics, producing a cured cell line. Here, we analyzed the glycoconjugate profiles of endosymbiont-harboring and cured strains of B. culicis and C. deanei by Western blotting and flow cytometry analyses using lectins that recognize specifically sialic acid and mannose-like residues. The absence of the endosymbiont increased the intensity of the lectins binding on both trypanosomatids. In addition, wild and cured strain-specific glycoconjugate bands were identified. The role of the surface saccharide residues on the interaction with explanted guts from Aedes aegypti gut was assessed. The aposymbiotic strains of B. culicis and C. deanei presented interaction rates 3.3- and 2.3-fold lower with the insect gut, respectively, when compared with the endosymbiont-bearing strains. The interaction rate of sialidase-treated cells of the wild and cured strains of B. culicis and C. deanei was reduced in at least 90% in relation to the control. The interaction of B. culicis (wild strain) with explanted guts was inhibited in the presence of mucin (56%), fetuin (62%), sialyllactose (64%) and alpha-methyl-D-mannoside (80%), while in C. deanei (wild strain) the inhibition was 53%, 56%, 79% and 34%, respectively. Collectively, our results suggest a possible involvement of sialomolecules and mannose-rich glycoconjugates in the interaction between insect trypanosomatids and the invertebrate host. PMID- 16216440 TI - Hydrogenase genes are uncommon and highly conserved in Rhizobium leguminosarum bv. viciae. AB - A screening for hydrogen uptake (hup) genes in Rhizobium leguminosarum bv. viciae isolates from different locations within Spain identified no Hup+ strains, confirming the scarcity of the Hup trait in R. leguminosarum. However, five new Hup+ strains were isolated from Ni-rich soils from Italy and Germany. The hup gene variability was studied in these strains and in six available strains isolated from North America. Sequence analysis of three regions within the hup cluster showed an unusually high conservation among strains, with only 0.5-0.6% polymorphic sites, suggesting that R. leguminosarum acquired hup genes de novo in a very recent event. PMID- 16216442 TI - Detection of Vibrio parahaemolyticus in cockle (Anadara granosa) by PCR. AB - This study aimed to determine the occurrence of Vibrio parahaemolyticus in cockles (Anadara granosa) at a harvesting area and to detect the presence of virulent strains carrying the thermostable direct hemolysin (tdh) and TDH-related hemolysin genes (trh) using PCR. Of 100 samples, 62 were positive for the presence of V. parahaemolyticus with an MPN (most probable number) value greater than 3.0 (>1100 MPN per g). The PCR analysis revealed 2 samples to be positive for the tdh gene and 11 to be positive for the trh gene. Hence, these results demonstrate the presence of pathogenic V. parahaemolyticus in cockles harvested in the study area and reveal the potential risk of illness associated with their consumption. PMID- 16216443 TI - Implication of hypR in the virulence and oxidative stress response of Enterococcus faecalis. AB - HypR has recently been described as the first transcriptional regulator involved in the oxidative stress response and in the intracellular survival of Enterococcus faecalis within macrophages. In order to characterize the HypR regulon, real-time quantitative RT-PCR experiments were performed. The expression of four genes involved in the oxidative stress response encoding catalase, glutathione reductase, and the two subunits of alkyl hydroperoxide reductase were down regulated in the hypR background under H(2)O(2) condition. These findings show that HypR acts as a transcriptional activator, especially during oxidative stress. In addition, DNAse I footprinting assays allowed us to identify the HypR protected DNA regions corresponding to the "HypR box" in the hypR promoter. Moreover, the effect of the hypR mutation on the virulence of E. faecalis was evaluated in comparison with the wild-type JH2-2 strain using a mouse peritonitis model. Our results revealed that HypR appears to be an important virulence factor in E. faecalis. PMID- 16216444 TI - Type III secretion: the bacteria-eukaryotic cell express. AB - Type III secretion (T3S) is an export pathway used by Gram-negative pathogenic bacteria to inject bacterial proteins into the cytosol of eukaryotic host cells. This pathway is characterized by (i) a secretion nanomachine related to the bacterial flagellum, but usually topped by a stiff needle-like structure; (ii) the assembly in the eukaryotic cell membrane of a translocation pore formed by T3S substrates; (iii) a non-cleavable N-terminal secretion signal; (iv) T3S chaperones, assisting the secretion of some substrates; (v) a control mechanism ensuring protein delivery at the right place and time. Here, we review these different aspects focusing in open questions that promise exciting findings in the near future. PMID- 16216445 TI - Small proteins of plant-pathogenic fungi secreted during host colonization. AB - Small proteins secreted by plant pathogenic fungi in their hosts have been implicated in disease symptom development as well as in R-gene mediated disease resistance. Characteristically, this class of proteins shows very limited phylogenetic distribution, possibly due to accelerated evolution stimulated by plant-pathogen arms races. Partly due to lack of clues from primary sequences, insight into the biochemical functions or molecular targets of these proteins has been slow to emerge. However, for some proteins important progress has recently been made in this direction. Expression of the genes for small secreted proteins is in many cases specifically induced after infection, which should help to advance our still very limited understanding of how plant pathogens recognize and respond to the host environment. PMID- 16216446 TI - A new soluble 10kDa monoheme cytochrome c-552 from the anammox bacterium Candidatus "Kuenenia stuttgartiensis". AB - The chemolithoautotrophic anammox bacterium Candidatus "Kuenenia stuttgartiensis" grows anaerobically using ammonium as electron donor for nitrite reduction. More than 10% of the proteins in cell extracts of "K. stuttgartiensis" consist of c type heme proteins. A 10kDa soluble cytochrome c was purified from cell extracts using ultracentrifugation and anion exchange chromatography. The UV/Vis spectrum of the reduced cytochrome showed the gamma, beta and alpha absorption maxima at 419, 522 and 552nm, respectively. The N-terminal amino acid sequence and peptide fragments of the tryptic digest of the protein were used to identify the corresponding gene. Analysis of the gene product showed that the protein was preceded by a 30 amino acids long leader sequence and that it belonged to the low spin class ID cytochrome c. The CXXCH motive was located at the N-terminal site of the protein. The gene organization of the cytochrome showed some resemblance to cytochrome c clusters of unknown function in the genome of Nitrosomonas europaea and Geobacter sulfurreducens PCA. PMID- 16216447 TI - Valosine-containing proteins (VCP) in an annelid: identification of a novel spermatogenesis related factor. AB - Two cDNAs similar to mammalian valosine-containing proteins (VCPs) were isolated from the common lumbricid earthworm Eisenia fetida (Savigny, 1826). The primary sequences, referred to as eVCP-1 and eVCP-2, display a similarity of 74%. Despite of the variable C-termini, both eVCPs have a conserved intron/exon organization spanning 14 kb, which is also conserved to their mammalian counterparts. Although this finding strongly suggests VCPs have a common ancestral origin, phylogenetic analysis predicts that eVCP-2 may be distinct. An investigation by reverse transcription-polymerase chain reaction (RT-PCR) revealed that, whilst evcp-1 was ubiquitously expressed during all developmental stages, evcp-2 was specifically expressed in the anterior segments of sexually mature earthworms. In situ hybridization clearly demonstrated that evcp-2 is expressed in the seminal vesicles, the location of spermatogenesis, and more precisely within the cytophores surrounded by secondary spermatocytes or spermatids. Taken together, this evidence leads to the notion that eVCP-2 is a likely component involved in the final modulation of spermatogenesis. PMID- 16216448 TI - Characterization of a complex genomic alteration on chromosome 2p that leads to four alternatively spliced fusion transcripts in the neuroblastoma cell lines IMR 5, IMR-5/75 and IMR-32. AB - Genetic aberrations in neuroblastoma (NB) have been extensively characterized over the last years. Alterations of the short arm of chromosome 2 (2p) have been of particular interest, since amplification of the MYCN oncogene on 2p24 is associated with an adverse outcome in NB patients. Here, we report on the characterization of a novel genomic rearrangement involving genetic material from 2p13 and 2p24 in NB cell lines that was discovered based on a serial analysis of gene expression (SAGE) profile of the MYCN-amplified NB cell line IMR-5. By analysis of a highly expressed SAGE tag not matching a Unigene cluster we identified four alternatively spliced corresponding transcripts, each of which consisted of the first 14 exons of the anthrax toxin receptor 1 gene (2p13.1) and varying combinations of exons of an unidentified gene located 1.3 Mb telomeric of MYCN (2p24.3) that was termed novel neuroblastoma gene 1. By Southern Blotting, Fluorescent In Situ Hybridization and Long Distance Inverse-PCR we disclosed that these transcripts result from a genomic alteration including material from distinct regions of chromosome 2p and four genomic breakpoints that are joined by short sequences of unknown origin. Furthermore, we show that this rearrangement lies within the homogeneous staining regions (HSR) in IMR-32 cells and is prevalent in both IMR-32 cells and their sub-clones IMR-5 and IMR-5/75, but not in a panel of 70 primary NB tumors. Our work is the first study discovering a fusion transcript based on a SAGE profile and for the first time precisely describes the DNA sequence of amplified breakpoint regions in NB. PMID- 16216449 TI - Gene duplication and functional divergence during evolution of the cytoskeletal linker protein talin. AB - The animal talins are large, modular proteins that link the actin cytoskeleton to the extracellular environment through interactions with beta-integrins and actin. Dictyostelium discoideum has two talins, TalA and TalB, which have distinct physiological roles in cell adhesion, cell differentiation, and cytokinesis. We previously identified a second talin gene in vertebrates. Thus, talin function in vertebrates is also due to the action of multiple proteins. Using a phylogenomic approach we have determined that D. discoideum TalA/B and the animal talins are related by descent from a common ancestral talin and that duplication of TLN2 early in the chordate lineage produced TLN1. An additional duplication subsequently produced a second Talin-2 in teleost fishes and a second Talin-1 in Xenopus laevis. We also show that vertebrate Talin-2 mRNA is alternatively processed. In the invertebrate Drosophila melanogaster and in the non-vertebrate chordate Ciona intestinalis, which each have only one talin gene, alternative processing of talin mRNA also produces multiple talin species. Thus, in these organisms, talin function may be due to the action of more than one protein. To identify isoform-specific functions of vertebrate talins we have shown through proteomic analysis that mammalian Talin-1 and Talin-2 bind to different protein partners. Further characterization of the differences between animal talins, especially the direct comparison of talins in the model urochordate C. intestinalis, which has one talin gene that produces two talins through alternative mRNA splicing, with Talin-1 and Talin-2 in model vertebrates, will provide an experimental system for studying neofunctionalization or subfunctionalization of talin following the vertebrate talin gene duplication. PMID- 16216450 TI - Sequence divergence and conservation in organizationally distinct subfamilies of Donax trunculus satellite DNA. AB - Characterization of a low-copy number DTF1 satellite DNA detected in the bivalve mollusk Donax trunculus revealed extensive grouping of monomer sequence variants into subfamilies identified by distinctive combinations of diagnostic nucleotides. It can be anticipated that a large number of subfamilies exists in the genome. In addition to the tandem organization of 169 bp long monomers, at least one subfamily was created through amplification of adjacent repeats in a higher order register. This complex satellite unit consists of two distinctive monomer variants that differ both in specific nucleotide changes and in a deleted segment partially substituted with a short unrelated sequence element. Most of the nucleotide substitutions differing between subfamilies are highly homogenized within a corresponding group of monomer variants, and intra-subfamily variability in general is low. Nucleotide diversity analysis of all sequenced variants of DTF1 satellite revealed the presence of two conserved segments, while the rest of the monomer sequence shows uniform and considerably higher level of variability. The persistence of conserved segments stands in contrast to the sequence and organizational divergence of monomer variant groups, and may indicate constraints in the evolution of DTF1 satellite repeats. PMID- 16216451 TI - The fungal biopesticide Metarhizium anisopliae has an adjuvant effect on the allergic response to ovalbumin in mice. AB - The parasitic fungus, Metarhizium anisopliae, is non-pathogenic to humans and licensed for indoor control of cockroach infestation. An important reason for the elimination of this vermin is that sensitisation to cockroaches is associated with asthma. Previously M. anisopliae has been shown to cause allergic- and asthma-like responses in mice and in the present study we have examined the adjuvant activity of M. anisopliae on the allergic response to the model allergen ovalbumin (OVA) in a mouse model. Levels of OVA-specific IgE, IgG1 and IgG2a in serum were measured and the weight and cell number of the excised popliteal lymph node were determined. Mice primed with mycelium+OVA and boosted with OVA had increased anti-OVA IgE and IgG1 levels compared with mice primed with OVA alone or mycelium. Priming with M. anisopliae (as mycelium or MACA) increased weight or cell number of the excised PLNs. These results suggest that M. anisopliae has the ability to increase an allergic response to an allergen and consequently, may worsen allergy in susceptible individuals. PMID- 16216452 TI - Effects of dimethylsulphoxide on mice arsenite-induced dysmorphogenesis in embryo culture and cytotoxicity in embryo cells. AB - Dimethylsulphoxide (DMSO) is a widely used vehicle for water insoluble compounds in experimental studies. Nevertheless, little is known about its potential impact on dysmorphogenesis caused by reactive oxygen species (ROS). In order to evaluate if DMSO at concentrations used as vehicle can alter in vitro sodium arsenite (Asi) teratogenicity and cytotoxicity, mouse embryos with 4-5 somites were grown for 48 h in Asi 0.4 and 4 microM, with and without 0.1% DMSO (v/v). Also embryonic mesenchymal cell were cultured, using mesenchymal mouse embryo cells obtained from gestation day 11 and treated with DMSO 0.1%, 0.2% and 0.5% (v/v) 15 min before Asi was added at final concentrations of 0.4 and 4 microM. Cytotoxicity and intracellular ROS production, were evaluated with MTT and 5-(and 6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM H2DCFDA), respectively. Results indicated that Asi produced growth retardation and abnormal development. Main malformations involved neural tube closure defects, abnormal rotation and optic vesicle defects among others. Co-treatment with DMSO partially reduced neural tube defects as well as facial dysmophology. Asi reduced cell viability inversely to the level of ROS production, and DMSO returned cellular viability to control values by reducing ROS intracellular production. In summary, the protective effect observed for DMSO appeared to reflect free radical scavenger properties, though other mechanisms independent to ROS production may have also been involved. PMID- 16216453 TI - Behavioral changes in metallothionein-null mice after the cessation of long-term, low-level exposure to mercury vapor. AB - The neurobehavioral changes in wild-type and metallothionein (MT)-null mice after the cessation of long-term, low-level exposure to Hg0 were investigated. MT-null and wild-type females were continuously (24 h/day) exposed to mercury vapor (Hg0) at 0.055 mg/m3 (range: 0.043-0.073 mg/m3), which was similar to the current threshold limit value (TLV), for 29 weeks. The effects on behavior, such as locomotor activity in the open field (OPF), learning ability in the passive avoidance response (PA) and spatial learning ability in the Morris water maze (MM) were examined immediately and 12 weeks after the cessation of exposure. Immediately after the exposure had ceased, total locomotor activity in OPF was decreased in the both strain of mice, although the MT-null mice appeared to show more distinct effect. In the PA test, the exposed animals of both strains showed learning impairment as compared to un-exposed mice. Twelve weeks after the cessation of exposure, the locomotor activity in OPF was elevated in the exposed mice of both strains, while the learning ability in the PA test appeared normal in both strains. Spatial learning ability was not affected at all. Immediately after the exposure had ceased, the brain mercury concentration of the exposed wild-type mice was 1.75 microg/g, twofold of that in the MT-null mice. In 12 weeks, brain mercury levels decreased to approximately 1/20 of those in immediately after the exposure in both of the strains. These results for the first time indicated that long-term, low-level exposure to Hg0 could exert neurobehavioral effects, which were not reversible even after a long exposure free period. Whereas the effects on learning ability were presumably transient, the effects on spontaneous behavior as evaluated in OPF were persistent. Finally, the MT-null mice seemed more susceptible to Hg0-induced neurotoxicity than the wild-type mice, confirming our previous results. PMID- 16216454 TI - Effect of Cistus laurifolius L. leaf extracts and flavonoids on acetaminophen induced hepatotoxicity in mice. AB - In this study, the effect of the flavonoids quercetin-3-methyl-ether (isorhamnetin) (1), quercetin-3,7-dimethyl-ether (2) and kaempferol-3,7-dimethyl ether (3) isolated from Cistus laurifolius L. (cistaceae) leaves was assessed on lipid peroxidation (liver and plasma), cellular glutathione (GSH) level and plasma AST (aspartate aminotransferase), ALT (alanine aminotransferase) enzyme activities in acetaminophen-induced liver damage in mice. At 114 mg/kg oral dose quercetin-3,7-dimethyl-ether was shown to possess potent antioxidative activity. PMID- 16216455 TI - Preliminary study on immunological and behavioural effects of Thymus broussonetii Boiss., an endemic species in Morocco. AB - In the present work, we had tried to evaluate the immunotropic and behavioural effects of Thymus broussonetii Boiss. So, we tested the neurostimulant effects of four extracts. This preliminary study allowed to identify both the immunostimulant and the neurotropic antistress effects of the studied extracts. Among the four extracts, only the aqueous and ethyl acetate ones showed an apparent effect on the tested biological activities, whereas the butanolic extract and the essential oil did not show any interesting effect (data not shown). These results showed that the aqueous and ethyl extracts of this endemic species are of interest for two reasons: stimulation of the immunizing system and protection against the stress by a neurotropic activity. Thyme extracts increased in vivo the number of leucocyte categories studied including polynuclears, total lymphocytes, TCD4+, TCD8+ and NK cells. These data suggest that the intraperitoneal administration of Thymus broussonetii extract has a potent direct effect on leucocytes in vivo. The elevation of leucocyte and thrombocyte counts produced by thyme in the peripheral blood was already reported in the literature. These results could be of practical importance in the field of phytotherapy in the treatment of some cases of human immunodeficiency such as cancer, leukaemia and AIDS. PMID- 16216457 TI - Medicinal efficacy of plants utilized as temple food in traditional Korean Buddhism. AB - We investigated the medicinal efficacies of plants used as food in 27 Korean Buddhist temples from 1997 to 2002. We studied 161 species of plants belonging to 135 genera in 65 families. Twenty-one plant parts were utilized as food in 42 different preparations. Approximately 82% of the plants studied had medicinal effects, with a wide range of efficacies (126 types). Of the medicinal plants, 52% were used for digestive problems, circulatory illnesses, and respiratory diseases. These results demonstrate that a high proportion of the food consumed in Korean temples is medicinal, and is used for a wide variety of diseases. PMID- 16216456 TI - Qualea grandiflora, a Brazilian "Cerrado" medicinal plant presents an important antiulcer activity. AB - Qualea grandiflora is one of the species widely used in folk medicine to treat gastric ulcers in Cerrado of the central region of Brazil. The hydroalcoholic extract of bark (HE) of Qualea grandiflora was investigated for their ability to prevent and heal lesions in the gastric mucosa. The oral administration of HE exhibited antiulcer activity decreasing the ulcerative index induced by HCl/ethanol solution, indomethacin/bethanechol and stress. In the Shay model, results showed that HE (p.o.) only reduced the severity of gastric lesions without effects on pH, gastric acidity or volume. When given by intraduodenal route, HE changed the pH, but did not modify the other parameters of the gastric juice. These data were in accordance with those obtained when HE was administered orally for 14 days after gastric ulcers were induced by acetic acid in rats. HE presented healing process in subacute gastric ulcer induced by acetic acid in rats. Moreover, histological examinations showed the simple columnar epithelium, lamina propria with simple branched tubular glandules with dilated lumen and large amounts of mucus secretion. Phytochemical investigation of HE led to the detection of terpenes, steroids, saponins, phenolic compounds and tannins in this extract, which may be involved in the observed activity. PMID- 16216458 TI - Development of latent fingerprints on compact disc and its effect on subsequent data recovery. AB - Chance fingerprints may be found on every type of surfaces of contact and when they are latent, need to be developed by various methods. The type of surface on which latent prints are to be developed is one of the important factors when a choice for a method of development is to be made. The matter becomes more crucial when the surface is unique like a compact disc containing digital data. In this case, to develop the fingerprints is not the only matter to be taken care of but also, is very important to select such a method which may not effect the stored data and its retrieval. In present investigation, various methods have been tried to develop fingerprints on the writing surface of a CD and results are discussed with respect to their development as well as its effect on stored data and data retrieval. PMID- 16216459 TI - Analysis of mutation of the plasma cholinesterase gene in a man who had died following a traffic accident. AB - We analyzed mutation of the butyrylcholinesterase (BCHE) gene in a 69-year-old man on whom a forensic autopsy had been performed after he had died following a traffic accident. Extremely low plasma cholinesterase activity had been pointed out by the emergency doctor at the hospital prior to his death and based on this, organophosphorus poisoning had been suspected. However, no pesticides, which could have reduced the plasma cholinesterase activity, were detected by toxicological analysis using GC/MS. Subsequently, one base insertion was found in exon 2. The frame shift mutation had occurred because a homozygous extra T had been inserted between nucleotides 1343 and 1344, resulting in the appearance of a stop codon in codon 454 (AGA454TAA, Arg454stop). This heterozygous frame shift mutation at this point was identified in the man's son. It is likely that there may be many such latent patients with abnormal plasma cholinesterase activity, and accordingly we should always bear this fact in mind and should carry out molecular genetic testing for an accurate diagnosis of this deficiency. PMID- 16216460 TI - A comparison of efficiency of manual and automatic fibres search with the Maxcan fibre finder. AB - The aim of this work was to study the efficiency of automatic fibre searching with the Maxcan fibre finder (Cox Analytical Systems, Sweden) in comparison to manual searching. The influence of some parameters (color, thickness, background noise) on the results of a fibre search was considered. Eighteen experimental tapes with different target fibres and different background noises were prepared in the laboratory. Searching of fibres was performed manually and with the Maxcan fibre finder by different operators from four European laboratories. Two laboratories have the Maxcan fibre finder system and the two instruments were used and compared in this study. The results show that searching with the Maxcan is generally as efficient as manual searching, except for very pale or very dark fibres. Note that the tapes used for these experiments were prepared in laboratory, and are not completely representative of the tape that could be obtained in real cases. To generalize the results obtained, further research on real case samples would be necessary. PMID- 16216461 TI - Perinatal programming of obesity. PMID- 16216462 TI - An investigation of DNA mismatch repair capacity under normal culture conditions and under conditions of supra-physiological challenge in human CD4+T cell clones from donors of different ages. AB - T cells undergo rapid clonal expansion upon antigenic stimulation to produce an effective immune response. Any defect in the DNA mismatch repair (MMR) system may have a detrimental effect on T cell proliferation. This study employed an in vitro model of human CD4+T cell ageing to investigate MMR capacity at various stages of T cell lifespan. A novel modification of the alkaline comet assay, which utilised T4 endonuclease VII to detect single base DNA mismatches, was used to assess DNA mismatch frequency. No clear pattern in DNA mismatch frequency with increasing culture age was observed. However, the ability to repair induced DNA mismatches (following treatment with acridine mutagen ICR-191) revealed an age related decline in the efficiency of the MMR system in clones derived from a 26 and a 45-year-old donor, but not from an 80-year-old very healthy SENIEUR donor. This study suggests that unchallenged, dividing human T cell clones have variable levels of DNA mismatches throughout their lifespan, not affecting proliferation. However, when challenged with supra-physiological levels of DNA mismatches, deficiencies were found in ageing T cell clones in MMR capacity, with the exception of T cell clones from a SENIEUR donor previously shown to maintain effective DNA excision repair. PMID- 16216463 TI - Genotoxicity and oxidative stress of the mutagenic compounds formed in fumes of heated soybean oil, sunflower oil and lard. AB - This study was to investigate the genotoxicity and cytotoxicity of the oil fumes formed from heating three common commercial cooking oils (soybean oil, sunflower oil, and lard) on human lung carcinoma pulmonary type II-like epithelium cell (A 549 cell). The major alkenal mutagenic compounds (trans-trans-2,4-decadienal, t-t 2,4-DDE; trans-trans-2,4-nonadienal, t-t-2,4-NDE; trans-2-decenal, t-2-DCA and trans-2-undecenal, t-2-UDA) contained in three oil fumes and their effects on the induction of reactive oxygen species (ROS) were also studied. It was found that the most potent mutagenic compound (t-t-2,4-DDE) of oil fumes was 66.4, 35.9 and 40.3 microg/g in soybean oil, sunflower oil and lard, respectively. The results indicated that the methanolic extracts of oil fumes could apparently lead to cytotoxicity and oxidative DNA damage. Glutathione (GSH) contents and the activities of antioxidant enzymes such as GSH reductase, and GSH S-transferase were adversely reduced by the methanolic extracts of oil fumes. When human A-549 cells were exposed to the methanolic extracts of oil fumes for 30 min, there was an increase in the formation of intracellular ROS, which was determined by dichlorofluorescein assay. Moreover, the methanolic extracts of oil fumes caused significant (p<0.05) oxidative damage through the 8-hydroxy-2'-deoxyguanosine formation in A-549 cells at the concentrations from 50 to 200 microg/ml. These results demonstrated that the DNA damage in A-549 cells, induced by cooking oil fumes, was related to the ROS formation. It is inferred that women exposed to emitted fumes from cooking oil were at higher risk of contracting lung cancer. PMID- 16216464 TI - Age-related decline in Digit-Symbol performance: eye-movement and video analysis. AB - Examines age-related decline in Digit-Symbol performance using variables obtained from a slow-motion analysis of a first person perspective video filmed during test completion, including superimposed cross-hairs indicating eye movements. Standard WAIS-3 DSCT scores and the video-derived variables were compared across two age groups (mean age 20 years vs. mean age 59 years). The older group performed more poorly overall, t(16)=-2.359, p=.031. The correlation between writing time per item and overall performance was (negatively) larger in the older group compared with the younger group, z=-2.180, p=.014. There was no difference between the groups' correlation coefficients with respect to key search latency and overall performance, z=-0.064, p=.525. Overall these results suggest that characterisation of the age-related slowing on Digit-Symbol tests as a psychomotor deficit is appropriate. PMID- 16216465 TI - Reliable change scores and their relation to perceived change in memory: implications for the diagnosis of mild cognitive impairment. AB - The relation between the subjective report of memory problems and objective evidence of the same has been debated with mixed results appearing in the literature. Less is known about the relation between objective change in test performance and the perceptions of cognitive change from family members/friends and trained clinicians. These relations were explored using 5-year longitudinal data from the population-based Canadian Study of Health and Aging. Statistically reliable deterioration in memory test performance was determined using a standardized regression-based (SRB) approach and a Reliable Change Index (RCI) that accounts for aging and practice effects. Among a subsample of persons with no cognitive impairment (NCI) at baseline, there was a moderate relation between reliable test score decline and ratings made by clinicians and informants. No relation, however, was found with the subjective reports of memory difficulties. These findings hold implications for current mild cognitive impairment (MCI) criteria which include subjective, informant and/or clinician ratings of cognitive decline. PMID- 16216466 TI - The placenta growth factor in skin angiogenesis. AB - The placenta growth factor (PlGF) is a member of the vascular endothelial growth factor (VEGF) family that has been shown to play an important role in promoting adult pathological angiogenesis. Besides inducing its own signaling in endothelial cells, PlGF exerts its angiogenic action by synergising with VEGF. In the skin, PlGF expression is upregulated during wound healing and PlGF-deficient mice show delayed wound closure, indicating that this factor promotes angiogenesis during skin repair. Moreover, PlGF expression by melanoma cells has been linked to tumor growth. The analysis of a transgenic mouse model constitutively expressing high levels of PlGF in basal keratinocytes has shown that this factor has strong angiogenic properties in the skin during both embryonic and post-natal life. Furthermore, PlGF delivery to the skin via an adenoviral vector induces the formation of large and stable blood vessels, but contrary to VEGF application, does not affect lymphatic vessel functionality. Such evidence opens the possibility of employing PlGF for therapeutic modulation of skin angiogenesis. PMID- 16216468 TI - Antibacterial activity of carbapenems against clinically isolated respiratory bacterial pathogens in Japan between 2003 and 2004. PMID- 16216467 TI - Microbiological rationale for the utilisation of prulifloxacin, a new fluoroquinolone, in the eradication of serious infections caused by Pseudomonas aeruginosa. AB - Minimal inhibitory concentrations (MICs) of prulifloxacin were evaluated in comparison with ciprofloxacin, levofloxacin and moxifloxacin against a large collection (N = 300) of Pseudomonas aeruginosa strains characterised according to the CLSI/NCCLS microdilution method. Additional in vitro tests (time-kill curves and mutant prevention concentration (MPC) determinations) were carried out. Assuming a susceptibility breakpoint for prulifloxacin identical to that of ciprofloxacin, the new fluoroquinolone emerged as the most potent antibiotic (72% of susceptible strains versus 65%, 61% and 23% for ciprofloxacin, levofloxacin and moxifloxacin, respectively). Time-kill tests at 4x MIC confirmed the pronounced bactericidal potency of the drug against P. aeruginosa. Amongst the members of the fluoroquinolone class assessed, prulifloxacin produced the lowest MPC values (< or = 4 mg/L). Our in vitro results indicate that prulifloxacin represents the most powerful antipseudomonal drug available today. PMID- 16216470 TI - Time-kill curves as a tool for targeting ceftazidime serum concentration during continuous infusion for treatment of septicaemic melioidosis. AB - Melioidosis is a fatal community-acquired infection endemic in tropical areas. Ten isolates of the causative microorganism were subjected to time-kill study using a range of ceftazidime concentrations. This study demonstrated that a ceftazidime concentration of eight times the minimum inhibitory concentration yielded an optimal bactericidal effect and should be the target concentration administered by continuous infusion. PMID- 16216469 TI - Cefotaxime and ceftriaxone cerebrospinal fluid levels during treatment of bacterial meningitis in children. AB - Cefotaxime (CTX) and ceftriaxone (CRO) were compared for cerebrospinal fluid (CSF) penetration and antimicrobial efficacy in cases of bacterial meningitis in children. This was a comparative study of CRO (100mg/kg once daily) and CTX (50 mg/kg 6 hourly) in the treatment of children with bacterial meningitis. The aetiological agents included Streptococcus pneumoniae (SPn), Haemophilus influenzae type b (Hib) and Neisseria meningitidis (NMen). Minimum inhibitory concentrations (MICs) were measured. In 33 patients from whom a second CSF specimen was obtained, CSF was cultured and assayed for antibiotic concentration. Median MICs of CTX and CRO for SPn, Hib and NMen were 0.01 and 0.01 microg/mL, 0.004 and 0.002 microg/mL and 0.008 and 0.004 microg/mL, respectively. All 33 repeat lumbar puncture specimens were sterile. The lowest CSF level recorded (0.45 microg/mL for CTX) was 45 times the MIC (0.01 microg/mL). The highest levels (24-35 microg/mL for CRO) were up to 8750 times the MIC of the patient's causative organism. A wide range of CSF levels for both antibiotics was observed. Levels varied with post-dose interval and duration of illness. On the basis of these findings, clinicians should be reassured that repeat lumbar puncture is not recommended for the causative organisms in this study (i.e., for Hib, NMen and penicillin/cefotaxime/ceftriaxone fully-susceptible SPn). PMID- 16216471 TI - Validation of the WHO Quality of Life assessment instrument (WHOQOL-100) in a population of Dutch adult psychiatric outpatients. AB - BACKGROUND: Research concerning the psychometric properties of the WHO Quality of Life Assessment Instrument (WHOQOL-100) in general populations of psychiatric outpatients has not been performed systematically. AIMS: To examine the content validity, construct validity, and reliability of the WHOQOL-100 in a general population of Dutch adult psychiatric outpatients. METHOD: A total of 533 psychiatric outpatients entered the study (438 randomly selected, 85 internally referred). Participants completed self-administered questionnaires for measuring quality of life (WHOQOL-100), psychopathological symptoms (SCL-90), and perceived social support (PSSS). In addition, they underwent two semi-structured interviews in order to obtain Axis-I and Axis-II diagnoses, according to DSM-IV. RESULTS: The drop-out percentage was low (7.1%). Of the 24 facets of the WHOQOL-100, 22 had a good distribution of scores, leaving out the facets physical environment and transport. Exploratory factor analysis revealed a four-factor structure, which was similar to earlier findings in patients with specific somatic diseases and depressive disorders. Various-a priori expected-positive and negative correlations were found between facets and domains of the WHOQOL-100, and dimensions of the SCL-90 and the PSSS-score, indicating good construct validity of the WHOQOL-100. The internal consistency of all facets and the four domains of the WHOQOL-100 was good (Cronbach's alpha's ranging from 0.62 to 0.93 and 0.64 to 0.84, respectively). Sparse and relatively low correlations were found between demographic characteristics (age and sex) and WHOQOL-100 scores. CONCLUSIONS: Content validity, construct validity, and reliability of the WHOQOL-100 in a population of adult Dutch psychiatric outpatients are good. The WHOQOL-100 appears to be a suitable instrument for measuring quality of life in adult psychiatric outpatients. PMID- 16216472 TI - Phospholipid transfer protein activity is associated with inflammatory markers in patients with cardiovascular disease. AB - Plasma phospholipid lipid transfer protein (PLTP) has several known key functions in lipoprotein metabolism. Recent studies suggest that it also may play a role in the inflammatory response. Inflammatory cell activity contributes to the development of atherosclerosis. To seek further evidence for the association of PLTP with inflammation, we studied the relationship between PLTP activity and five inflammatory markers [C-reactive protein (CRP), serum amyloid A (SAA), interleukin 6 (IL-6), white blood cells (WBC), and fibrinogen] in 93 patients with low HDL and cardiovascular disease (CVD). Plasma PLTP activity had the strongest correlation with CRP (r=0.332, P<0.001) followed by SAA (r=0.239, P=0.021). PLTP, CRP, and SAA were significantly associated with body mass index (BMI), insulin or glucose, apolipoprotein (apo) B, and/or apo E level (r=0.264 0.393, P<0.01). PLTP, SAA, and IL-6 also were associated with the concentration of HDL particles without apo A-II [Lp(A-I)](r=0.373-0.472, P<0.005, n=56), but not particles with apo A-II. Smoking was associated with increased PLTP activity, CRP, and WBC, and hypertension with increased PLTP activity. In linear models, CRP remained significantly associated with PLTP after adjustment of CVD risk factors and insulin resistance. Also, much of the variability of plasma PLTP activity was explained by CRP, BMI, Lp(A-I), smoking, glucose, and blood pressure. These findings show for the first time that plasma PLTP activity is associated positively with CRP in CVD, a state of chronic inflammation. PMID- 16216473 TI - HPRTSardinia: a new point mutation causing HPRT deficiency without Lesch-Nyhan disease. AB - Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency always causing hyperuricemia presents various degrees of neurological manifestations, the most severe which is Lesch-Nyhan syndrome. The HPRT gene is situated in the region Xq26-q27.2 and consists of 9 exons. At least 300 different mutations at different sites in the HPRT coding region from exon 1 to exon 9 have been identified. A new mutation in the HPRT gene has been determined in one patient with complete deficiency of erythrocyte activity, with hyperuricemia and gout but without Lesch Nyhan disease. Analysis of cultured fibroblasts revealed minimal residual HPRT activity mainly when guanine was the substrate. Genomic DNA sequencing demonstrated patient's mother heterozygosity for the mutation and no mutation in her brother. The mutation consists in a C-->T transversion at cDNA base 463 (C463T) in exon 6, resulting in proline to serine substitution at codon 155 (P155S). This mutation had not been reported previously and has been designated HPRT(Sardinia). The mutation identified in this patient allows some expression of functional enzyme in nucleated cells such as fibroblasts, indicating that such cell type may add further information to conventional blood analysis. A multicentre survey gathering patients with variant neurological forms could contribute to understand the pathophysiology of the neurobehavioral symptoms of HPRT deficiency. PMID- 16216474 TI - Frequency of hemochromatosis gene (HFE) mutations in Russian healthy women and patients with estrogen-dependent cancers. AB - Possible association between the C282Y and H63D mutations in the HFE gene and estrogen-dependent cancer risk was assessed. Genotyping was performed using PCR amplification followed by digestion of products with specific restrictases. In a population of 260 healthy women (permanent residents of the southwest European Russia), mutant allele frequencies at the C282Y and H63D sites were evaluated as 3.3 and 16.3%, respectively. In patients with breast, ovarian, and endometrial cancer, C282Y frequencies were also low (1.0, 1.3, and 3.8%, respectively), and no cancer risk associated with the C282Y mutation was found. Odds ratios for breast cancer risk associated with the H63D mutation increased significantly with age: 0.5 in women below 48 years old, 1.0 in a range of 48-57 years, and 4.4 in older women (P(trend)=0.002). The latter value was statistically significant (95% CI, 1.4-14.1), indicating that women bearing the H63D mutation may be at an increased breast cancer risk at an age above 57 years. Preliminary results obtained in patients with two other estrogen-dependent malignancies revealed the same tendency to OR increase with age in ovarian cancer patients (P(trend)=0.008), but no age-related OR differences in endometrial cancer patients. PMID- 16216475 TI - 93Nb NMR chemical shift scale for niobia systems. AB - 93Nb solid-state NMR spectra of a series of inorganic niobates with Nb in different oxygen coordination environments were measured. For all studied compounds the chemical shielding and quadrupole tensor parameters were determined using conventional and ultrahigh field NMR facilities, ultrahigh speed MAS, DQ STMAS, solid-echo and computer modeling. It has been demonstrated that the 93Nb isotropic chemical shift is sensitive to the coordination number of Nb sites. For the first time the 93Nb NMR chemical shift scale for NbOx polyhedra in solid materials has been proposed: for four-coordinated Nb sites, the isotropic shifts occur from -650 to -950 ppm; five-coordinated Nb sites have the isotropic shifts in the range of -900 to -980 ppm; for six-coordinated Nb sites the isotropic shifts vary from -900 to -1360 ppm; the shifts from -1200 to -1600 ppm are typical for seven-coordinated Nb sites; for eight-coordinated Nb sites the shifts are higher than -1400 ppm. The possible correlation between the value of the isotropic chemical shift and the ionic character of the NbOx-MOy polyhedra association has been suggested. The magnitude of the 93Nb quadrupole coupling constant depends on the local symmetry of Nb sites and may vary from hundreds of kHz to hundreds of MHz. PMID- 16216477 TI - Amplitude-modulated decoupling in rotating solids: a bimodal Floquet approach. AB - This paper centers on a theoretical study of amplitude-modulated heteronuclear decoupling in solid-state NMR under magic-angle spinning (MAS). A spin system with a single isolated rare spin coupled to a large number of abundant spins is used in the analysis. The phase-alternating decoupling scheme (XiX decoupling) is analyzed using bimodal Floquet theory and the operator-based perturbation method developed by van Vleck. An effective Hamiltonian correct to second order is calculated for the spin system under XiX decoupling. The results of these calculations indicate that under XiX decoupling the main contribution to the residual line width comes from a cross-term between the heteronuclear and the homonuclear dipolar couplings. This is in contrast to continuous-wave decoupling, where the residual line width is dominated by the cross-term between the heteronuclear dipolar coupling and the chemical-shielding tensor of the irradiated spin. For high-power decoupling the method results in very good decoupling provided that certain unfavorable recoupling conditions, imposed by specific ratios of the amplitude modulation frequency and the MAS frequency, are avoided. For low-power decoupling, the method leads to acceptable decoupling when the pulse length corresponds to an integer multiple of a 2pi rotation and the rf field amplitude is less than a quarter of the MAS frequency. The performance of the XiX scheme is analyzed over a range of values of the rf power, and numerical results that agree well with the most recent experimental observations are presented. PMID- 16216478 TI - Optic flow dominates visual scene polarity in causing adaptive modification of locomotor trajectory. AB - Locomotion and posture are influenced and controlled by vestibular, visual and somatosensory information. Optic flow and scene polarity are two characteristics of a visual scene that have been identified as being critical in how they affect perceived body orientation and self motion. The goal of this study was to determine the role of optic flow and visual scene polarity on adaptive modification in locomotor trajectory. An object is said to have visual polarity, or to be "visually polarized", when it contains an identifiable principal axis with one end distinct from the other. Two computer-generated virtual reality scenes were shown to subjects during 20 min of treadmill walking. One scene was a highly polarized scene, while the other was composed of objects displayed in a non-polarized fashion. Both virtual scenes depicted constant rate self motion equivalent to walking counterclockwise around the perimeter of a room. Subjects performed Stepping Tests blindfolded before and after scene exposure to assess adaptive changes in locomotor trajectory. Subjects showed a significant difference in heading direction, between pre- and post-adaptation Stepping Tests, when exposed to either scene during treadmill walking. However, there was no significant difference in the subjects' heading direction between the two visual scene polarity conditions. Therefore, it was inferred from these data that optic flow has a greater role than visual polarity in influencing adaptive locomotor function. PMID- 16216476 TI - Dynamic nuclear polarization and nuclear magnetic resonance in the vicinity of edge states of a 2DES in GaAs quantum wells. AB - Nuclear magnetic resonance is detected via the in-plane conductivity of a two dimensional electron system at unity Landau level filling factor in the regime of the quantum Hall effect in narrow and wide quantum wells. The NMR is spatially selective to nuclei with a coupling to electrons in the current carrying edge states at the perimeter of the 2DES. Interpretation of the electron-nuclear double resonance signals is facilitated by numerical simulations. A new RF swept method for conductivity-detected NMR is introduced which offers more efficient signal averaging. The method is applied to the study of electric quadrupole interactions, weakly allowed overtone transitions, and evaluation of the extent of electron wave function delocalization in the wide quantum well. PMID- 16216480 TI - [Radiological quiz of the month. Multiple neurological symptoms in a 14-year-old child]. PMID- 16216479 TI - Partial unilateral inactivation of the dorsal hippocampus impairs spatial memory in the MWM. AB - The hippocampus is one of the more widely studied structures related with spatial memory. In this study, we assessed the effect of unilateral inactivation of the dorsal hippocampus with tetrodotoxin (TTX) on the performance displayed by Wistar rats in the spatial version of the Morris water maze. In experiment 1, we injected into the dorsal hippocampus in two different groups of rats 1 microl of saline solution or 5 ng of TTX in 1 microl of saline each day immediately after the training during four consecutive days. This procedure blocked consolidation and impaired spatial memory in the TTX group. In experiment 2, a new group of subjects was trained in the Morris water maze for 8 days and was administered 1 microl of saline on day 7 (saline session) and TTX on day 8 (TTX session) into the dorsal hippocampus 40 min before the training. Only the treatment with TTX altered the retrieval of memories. These experiments showed that unilateral interventions on the dorsal hippocampus can affect consolidation as well as retrieval of well-established spatial memories. PMID- 16216481 TI - [Feeding practices in infants: a 6-month prospective cohort study]. AB - The aims of this work were to assess timing of complementary feeding in infants and to precise the underlying factors that may cause inappropriate complementary feeding. PATIENTS AND METHODS: A cohort of 167 newborns, born in the same maternity ward during a 6 week-period, was prospectively analyzed. Only healthy neonates were included in the study. A phone questionnaire was filled at 4 and 6 months of age to evaluate modality of complementary feeding. Multivariate analysis (segmentation tree, analyse by multiple correspondence) was used to study factors associated with inappropriate diversification. RESULTS: Out of the 167 neonates included in the study, 132 mothers could be contacted at 4 months and 116 at 6 months of age. Sixty-seven per cent of mothers started breast feeding at birth. Among these, 33% still breastfed -at least partially- at 4 months and 17% at 6 months. Fifty-two percent of mothers started complementary feeding before 4 months, and 24% of infants received gluten at 4 months of age. Multi-gravida mothers, mothers aged more than 35 years old and mothers who gave infant or follow-up formulae before 4 months, started complementary feeding significantly earlier (P<0.05). Infants who were formula fed received more frequently complementary feeding before the age of 4 months than breast fed infants (57% vs 33%, P<0.05). CONCLUSION: Our study showed that half of infants were introduced solid food too early and allowed to identify a population at risk that could benefit from nutritional intervention programs. PMID- 16216482 TI - [Quality of life of adolescents surviving childhood acute lymphoblastic leukemia]. AB - PURPOSE: To evaluate how adolescents and young adults cured of acute lymphoblastic leukemia (ALL) treated during childhood have integrated the disease, and possible death related to cancer. Particularly, we have focused on experiences related to diagnosis announcement, hospitalisation and treatments and consequences on their social, psychological and somatic behaviour. PATIENTS: Forty-one patients cured of ALL have been enrolled in the study and answered one interview with clinical psychologist or research nurse. RESULTS: Although 60% of the patients argued that they think rarely of their disease, 10% thought about it every day. Traumatic evidence was detectable in most of them. Physical pain was the most reported stress, mainly during hospitalisation (93%), as well as psychological suffering (83%). Afterwards, the mostly often-reported stress was psychological pain (61%). Sixty-six percent declared that they still experience psychological and health consequences at the time of the interview, in some cases reported as a handicap in their life. In 83% of the cases they considered themselves as cured, nevertheless fear of relapse persisted in 1/3. Ninety percent said they have a pleasant life, 56% did not like to talk about leukaemia and 70% thought they could have died. For 85%, disease has been the most important event of their life and 75% testify to repercussions of the disease on their family (family relationship changes, overprotection, siblings difficulties). CONCLUSION: Most of these patients declared to be 'as the others' and developed life projects, but overcoming the pain experience of the disease remained difficult. This study emphasized the need for long-term continuous information and reinforces the importance of addressing treatment psychological and physical pain mainly after the initial hospitalisation period. PMID- 16216483 TI - Eugenol--the active principle from cloves inhibits 5-lipoxygenase activity and leukotriene-C4 in human PMNL cells. AB - Polymorphonuclear leukocytes (PMNL) play an important role in the modulation of inflammatory conditions in humans. PMNL cells recruited at the site of inflammation, release inflammatory mediators such as leukotrienes, proteolytic enzymes and reactive oxygen species. Among these, leukotrienes are implicated in pathophysiology of allergic and inflammatory disorders like asthma, allergic rhinitis, arthritis, inflammatory bowel disease and psoriasis. 5-lipoxygenase (5 LO) is the key enzyme in biosynthetic pathway of leukotrienes. Our earlier studies showed that spice phenolic active principles significantly inhibit 5-LO enzyme in human PMNLs. In this study we have further characterized the inhibitory mechanism of eugenol, the active principle of spice-clove on 5-LO enzyme and also its effect on leukotriene C((4)) (LTC(4)). Substrate dependent enzyme kinetics showed that the inhibitory effect of eugenol on 5-LO was of a non-competitive nature. Further, eugenol was found to significantly inhibit the formation of LTC(4) in calcium ionophore A23187 and arachidonic acid (AA) stimulated PMNL cells. These data clearly suggest that eugenol inhibits 5-LO by non-competitive mechanism and also inhibits formation of LTC(4) in human PMNL cells and thus may have beneficial role in modulating 5-LO pathway in human PMNL cells. PMID- 16216484 TI - How robust is the evidence for viruses in the induction of type 1 diabetes? AB - Viruses associated with type 1 diabetes have eluded definition as causal, with the exception of rubella virus. False-negative results may have occurred due to the focus on subjects at symptomatic onset, who may be heterogeneous and differently affected by viruses. In addition, assays have not always been sufficiently sensitive to deal with transient infections, and pancreatic tissue is scarce. Longitudinal studies of at-risk subjects and more sensitive DNA techniques now reveal that at initiation of islet autoimmunity, enteroviruses have only a small role, but are more likely to be important at symptomatic onset. Rotaviruses remain associated with initiation of islet autoimmunity, and generate strong T cell responses in the young. PMID- 16216485 TI - In vitro evaluation of an asthma dosing device: the smart-inhaler. AB - Monitoring devices attached to pressurised metered dose inhalers provide an important objective measurement of patient adherence with asthma medications in clinical and research settings. The Smart-inhaler is a relatively new device that has not been previously validated. This study examines the accuracy of the Smart inhaler in a bench-top experiment and compares it with a previously validated device, the Doser. Ten Smart-inhalers and five Dosers were actuated twice on two occasions per day for 30 days (120 doses). Six Smart-inhalers were also actuated 30 times in rapid succession to examine the ability of the Smart-inhaler to detect "dumping". Five Smart-inhalers failed to detect the first one or two doses. However, when the aerosol canister was placed more firmly in the device, actuating the device in the process, the following two doses were recorded accurately in all ten devices. Otherwise all ten Smart-inhalers and five Dosers recorded all actuations faithfully and there were no spurious recordings. The six Smart-inhalers recorded all 30 doses delivered in rapid succession. The Smart inhaler and Doser are both highly accurate at measuring actuated doses and no spurious doses were recorded in an in vitro setting. PMID- 16216486 TI - Dietary gangliosides increase the content and molecular percentage of ether phospholipids containing 20:4n-6 and 22:6n-3 in weanling rat intestine. AB - This study was conducted to determine whether dietary ganglioside (GG) increases the content of ether phospholipids (EPL) in intestinal mucosa. Weanling Sprague Dawley rats were fed a semipurified diet consisting of 20% fat as a control diet. Two experimental diets were formulated by adding either 0.1% (w/w fat) GGs (GG diet) or 1.0% (w/w fat) sphingomyelin (SM diet) to the control diet. Fatty acid methyl esters from the alkenylacyl, alkylacyl and diacyl subclasses of phospholipids were measured to determine total and molecular percentage of EPL comprising the choline phosphoglyceride (CPG) and ethanolamine phosphoglyceride (EPG) fraction. Animals fed the GG diet significantly increased total EPL content both in CPG (by 36%) and in EPG (by 66%), and the molecular percentage of EPL in CPG (by 76%) and in EPG (by 59%) compared to animals fed the control diet. Dietary GG-induced increase in EPL resulted in a higher level of polyunsaturated fatty acids (PUFA) specifically in 20:4n-6 and 22:6n-3 compared to control animals, leading to a decrease in the ratio of saturated fatty acids (SFA) to PUFA both in CPG and in EPG. Feeding animals the SM diet showed a higher level of EPL than control animals with a concomitant increase in 22:6n-3 in EPL. The present data demonstrate that dietary GG increases the content and composition of EPL containing PUFA in the weanling rat intestine. PMID- 16216487 TI - Differential action of 13-HPODE on PPARalpha downstream genes in rat Fao and human HepG2 hepatoma cell lines. AB - In rats, oxidized fats activate the peroxisome proliferator-activated receptor alpha (PPARalpha), leading to reduced triglyceride concentrations in liver, plasma and very low density lipoproteins. Oxidation products of linoleic acid constitute an important portion of oxidized dietary fats. This study was conducted to check whether the primary lipid peroxidation product of linoleic acid, 13-hydroperoxy-9,11-octadecadienoic acid (13-HPODE), might be involved in the PPARalpha-activating effect of oxidized fats. Therefore, we examined the effect of 13-HPODE on the expression of PPARalpha target genes in the rat Fao and the human HepG2 hepatoma cell lines. In Fao cells, 13-HPODE increased the mRNA concentration of the PPARalpha target genes acyl-CoA oxidase (ACO), cytochrome P450 4A1 and carnitine-palmitoyltransferase 1A (CPT1A). Furthermore, the concentration of cellular and secreted triglycerides was reduced in Fao cells treated with 13-HPODE. Because PPARalpha mRNA was not influenced, we conclude that these effects are due to an activation of PPARalpha by 13-HPODE. In contrast, HepG2 cells seemed to be resistant to PPARalpha activation by 13-HPODE because no remarkable induction of the PPARalpha target genes ACO, CPT1A, mitochondrial HMG-CoA synthase and delta9-desaturase was observed. Consequently, cellular and secreted triglyceride levels were not changed after incubation of HepG2 cells with 13-HPODE. In conclusion, this study shows that 13-HPODE activates PPARalpha in rat Fao but not in human HepG2 hepatoma cells. PMID- 16216488 TI - A cowpea mosaic virus nanoscaffold for multiplexed antibody conjugation: application as an immunoassay tracer. AB - Cowpea mosaic virus (CPMV), an icosahedral 30 nm virus, offers a uniquely programmable biological nanoscaffold. This study reports initial optimization of the simultaneous modification of two CPMV mutants with AlexaFluor 647 fluorescent dyes and either IgG proteins or antibodies at specific sites on the virus scaffold. The capacity of CPMV as a simultaneous carrier for different types of molecules was demonstrated, specifically, when applied as a tracer in direct and sandwich immunoassays. The ability to label the virus capsid with antibody and up to 60 fluorescent dyes resulted in an improved limit of detection in SEB sandwich immunoassays, when used as a tracer, relative to a mole equivalent of dye-labeled antibody. PMID- 16216489 TI - Ancestral state reconstructions for genomes. AB - The recent expansion of phylogenetic analysis from the traditional field of molecular evolution, analyzing histories of genes, to the nascent field of "genomic evolution", analyzing histories of entire genomes, enables the construction of trees based on genome information, the quantification of the key processes that shape genome content and, ultimately, plausible parsimony reconstructions of ancestral genomes. Thus, when genomes are considered as phylogenetic characters, it is possible to reconstruct not only the history of species but also the ancestral states in terms of genome structure or function. In the future, we might be able to accurately reconstruct--or retrodict--a chain of events that led to the emergence of a specific genome sequence and, ultimately, to synthesize ancestral genomes at will, creating a "Jurassic database" of genomes. PMID- 16216490 TI - A comparative molecular modeling study of dydrogesterone with other progestational agents through theoretical calculations and nuclear magnetic resonance spectroscopy. AB - 6-Dehydroretroprogesterone (dydrogesterone) and three other natural or synthetic progestins (progesterone, retroprogesterone, and 6-dehydroprogesterone) were submitted to a conformational study through theoretical calculations at the B3LYP/6-31G(*) level and high field NMR spectroscopy. The study allows to define the role of the two structural features which differentiate these steroids, i.e., the C9 and C10 configuration and the C6-C7 unsaturation. The combined effects of the conformational preference of A ring, determined by the configuration at C9 and C10, and the enhanced rigidity due to the C6-C7 double bond, could account both for the higher activity and selectivity of dydrogesterone with respect to the other three steroids. PMID- 16216491 TI - Endocrinology of pregnancy: consequences for the diagnosis and treatment of pregnancy disorders. AB - Placental human chorionic gonadotropin and corpus luteum secretion of progesterone and oestradiol are the main endocrine events at the beginning of pregnancy, whilst the luteo-placental shift is an important step during the later stages. Progesterone not only affects decidualisation, but is the major immunological determinant and controls uterine contractibility and cervical competence. These properties all contribute considerably towards the correct development of pregnancy and delivery at term. PMID- 16216492 TI - An LXXLL motif in nuclear receptor corepressor mediates ligand-induced repression of the thyroid stimulating hormone-beta gene. AB - Nuclear receptor corepressor (N-CoR) regulates gene expression through interaction with DNA-bound nuclear receptors, recruiting multicomponent repressor complexes to the sites of target genes. We recently reported the presence of an LXXLL motif in N-CoR, and showed that this motif interacts in vitro and in vivo with retinoic acid receptor alpha (RARalpha) and thyroid hormone receptor beta (TRbeta). Transient transfection experiments now suggest that TRbeta and N-CoR act synergistically and may both be required for ligand-induced repression from the negative TR response element in the thyroid stimulating hormone-beta (TSHbeta) gene promoter. Mutation of the LXXLL motif in N-CoR abolished ligand induced repression at this response element. Furthermore, in vitro binding of N CoR to a complex between TRbeta and the negative TR response element was strictly ligand-dependent. We conclude that N-CoR and TRbeta cooperate in the regulation of the TSHbeta gene and that the ligand-dependent repression is mediated by the LXXLL motif in N-CoR. PMID- 16216493 TI - New anabolic steroid illegally used in cattle-structure elucidation of 19 norchlorotestosterone acetate metabolites in bovine urine. AB - 4-Chloro-estr-4-en-17-ol-3-one, trivially named 19-norclostebol acetate or 4 chloro-19-nortestosterone acetate (NClTA), has been identified on the European black market in the late 1990s for possible use in breeding animals. After oral and subcutaneous administration of NClTA to bovine, urine samples were collected over a period of three weeks, and chemical structure of main excreted urinary metabolites was determined. After oral administration, the most abundant metabolites were mainly reduced as 4-chloro-19-norandrostan-3xi-ol-17-one and 4 chloro-19-norandrostan-3xi,17xi-diol. They were identified until 1 week after administration. Following subcutaneous injection, 4-chloro-19-norandrostan-3xi-ol 17-one was again of major abundance, but so were 4-chloro-19-norandrost-4-ene 3xi,17xi-diol and 4-chloro-19-norandrost-4-en-3xi-ol-17-one. They were detected at least 3 weeks after administration. Whatever the route of administration, metabolites were found mainly glucurono-conjugated; the only exception was metabolite 4-chloro-19-norandrostan-3xi-ol-17-one which was identified both in the sulpho- and glucurono-fractions. PMID- 16216494 TI - The effect of microwave irradiation on enzymatic hydrolysis of rice straw. AB - A series of experiments involving microwave irradiation were carried out to evaluate the effect of microwave irradiation on enzymatic hydrolysis of rice straw. Compared with microwave irradiation free hydrolysis, rice straw pretreated by combining microwave irradiation with alkali could increase the initial hydrolysis rate but the hydrolysis yield remained unchanged. When the enzyme solution was treated by microwave irradiation, the initial hydrolysis rate increased slightly, but the yield was decreased remarkably. Its optimal hydrolysis conditions were temperature (45 degrees C), pH (4.8) and enzyme loading (20 mg g(-1) substrate), which was determined by an orthogonal experiment. When intermittent microwave irradiation was used, initial hydrolysis rate was greatly accelerated but the yield was decreased slightly. Its optimal hydrolysis conditions were temperature (50 degrees C), pH (4.8) and enzyme loading (20 mg g(-1) substrate), which was determined by another orthogonal experiment. PMID- 16216495 TI - Statistical optimization of thermo-tolerant xylanase activity from Amazon isolated Bacillus circulans on solid-state cultivation. AB - A 2(2) factorial design was performed to find the best conditions of pH and temperature for xylanolytic activity of Bacillus circulans BL53 isolated from the Amazon environment. Solid-state cultivation was carried out on an inexpensive, abundant agro-industrial soybean residue. The central composite design (CCD) used for the analysis of treatment combinations showed that a second-order polynomial regression model was in good agreement with experimental results, with R(2) = 0.9369 (P < 0.05). The maximum activity was obtained at a high temperature (80 degrees C) and over a large pH range (4.0-7.0). Enzymatic activity was maintained in heated extracts up to 50 degrees C, suggesting that the xylanases of B. circulans BL53 are thermo-tolerant biocatalysts, being of interest for industrial processes. The crude enzyme extract hydrolyzed rice straw, sugar cane bagasse and soybean fiber and its activity was stimulated by Co(2+), Fe(3+), and beta mercaptoethanol but inhibited by Mn(2+), Cu(2+), Ca(2+), Zn(2+), Ba(2+), Mg(2+) and by EDTA. PMID- 16216496 TI - Pulvinones as bacterial cell wall biosynthesis inhibitors. AB - Pulvinones were synthesized (>180) in arrays and evaluated as inhibitors of early stage cell wall biosynthesis enzymes MurA-MurD. Several pulvinones inhibited Mur enzymes with IC(50)'s in the 1-10 microg/mL range and demonstrated antibacterial activity against Gram-positive bacteria including methicillin-resistant Staphyloccus aureus, vancomycin-resistant Enterococcus faecalis, and penicillin resistant Streptococcus pneumoniae. PMID- 16216497 TI - Discovery of A-770041, a src-family selective orally active lck inhibitor that prevents organ allograft rejection. AB - We describe the identification, SAR, and pharmacology of the src-family selective lck inhibitor A-770041 that prolongs the survival of major histocompatibility mismatched allografts in models of solid organ transplant rejection for greater than 65 days. PMID- 16216498 TI - Screening of electrophilic compounds yields an aziridinyl peptide as new active site directed SARS-CoV main protease inhibitor. AB - The coronavirus main protease, M(pro), is considered a major target for drugs suitable to combat coronavirus infections including the severe acute respiratory syndrome (SARS). In this study, comprehensive HPLC- and FRET-substrate-based screenings of various electrophilic compounds were performed to identify potential M(pro) inhibitors. The data revealed that the coronaviral main protease is inhibited by aziridine- and oxirane-2-carboxylates. Among the trans-configured aziridine-2,3-dicarboxylates the Gly-Gly-containing peptide 2c was found to be the most potent inhibitor. PMID- 16216499 TI - Affinity identification of delta-opioid receptors using latex nanoparticles. AB - Three types of latex nanoparticles carrying naltrindole (NTI) derivatives were synthesized as probes for the affinity isolation of their binding proteins including the delta-opioid receptor. The effect of the attachment of NTI to different positions on the linker was investigated. Only latex nanoparticles in which the NTI derivative was linked through the phenol group were useful for isolating the recombinant delta-opioid receptor solubilized from CHO cell membrane. These latex nanoparticles could be a useful tool for investigations of the pharmacological activity of NTI. PMID- 16216500 TI - Synthesis and biological evaluation of novel heterocyclic quinones as inhibitors of the dual specificity protein phosphatase CDC25C. AB - A focused set of heterocyclic quinones based on the benzothiazole, benzoxazole, benzimidazole, indazole and isoindole was prepared and screened with respect to the inhibition of the phosphatase activity of CDC25C. Benzoxazole- and benzothiazole-diones were at least 50 times more potent in inhibiting CDC25C than their benzimidazole-indazole- or isoindole-dione counterparts. These in vitro activities were in good correlation with the anti-proliferative effects observed with Mia PaCa-2 and DU-145 human tumor cell cultures. The IC(50) values obtained by WST-1 colorimetric assay ranged from 0.10 to 0.50 microM for the benzoxazole- or benzothiazole-diones and were above 10 microM for the other heterocyclic diones. This study further illustrates how the activity of the quinone pharmacophore can be selectively modulated by changing the type of five-membered heterocycle fused to the quinone ring. PMID- 16216501 TI - New 7,8-ethylenedioxy-2,3-benzodiazepines as noncompetitive AMPA receptor antagonists. AB - A series of 1-aryl-3,5-dihydro-7,8-ethylenedioxy-4H-2,3-benzodiazepin-4-ones 2a f, were synthesized and screened as anticonvulsant agents in DBA/2 mice against sound-induced seizures. The new compounds display anticonvulsant properties although the ED(50) values are higher than those of prototypes 1-aryl-3,5-dihydro 7,8-methylenedioxy-4H-2,3-benzodiazepin-4-ones (1) and GYKI 52466, well-known noncompetitive AMPA receptor antagonists. Functional tests were performed to evaluate the antagonistic activity at the AMPA and kainate receptors. PMID- 16216502 TI - Scaffold oriented synthesis. Part 1: Design, preparation, and biological evaluation of thienopyrazoles as kinase inhibitors. AB - We report the synthesis of kinase targeted libraries based on the thienopyrazole scaffold. Several thienopyrazole analogs have been identified as submicromolar inhibitors of KDR. PMID- 16216503 TI - Synthesis of a novel biotin-tagged photoaffinity probe for VEGF receptor tyrosine kinases. AB - A novel biotin-tagged photoaffinity probe was synthesized and evaluated as a vascular endothelial growth factor receptor-2 (VEGFR-2) tyrosine kinase inhibitor. The probe (2) is a potent VEGFR-2 inhibitor with an IC(50) value of 7.1 microM, and inhibits VEGF-induced proliferation in human umbilical vein endothelial cells (HUVEC), with an IC(50) value of 40.3 microM. This probe will be a useful reagent for investigating ligand-protein interactions. PMID- 16216504 TI - Biological evaluation of 1-alkyl-3-phenylthioureas as orally active HDL-elevating agents. AB - A series of 1-alkyl-3-phenylthiourea analogues were prepared and evaluated as HDL and Apo A-I-elevating and triglyceride-lowering agents. Several derivatives were superior to gemfibrozil. The optimal analogue (HDL376) was shown to raise HDL cholesterol in the rat, hamster, dog, and monkey models. PMID- 16216505 TI - A novel class of potent influenza virus inhibitors: polysubstituted acylthiourea and its fused heterocycle derivatives. AB - A series of polysubstituted and fused heterocycle derivatives of acylthiourea was prepared and the biological activity against influenza virus was evaluated. Of the analogues that demonstrated IC(50)s<0.1 microM, acylthiourea derivatives 16 and 50 were further investigated as candidates with the most potential for future development. The SAR of these compounds are discussed and they represent a novel class of highly potent and selective inhibitors of influenza virus. PMID- 16216506 TI - 3-Arylpiperazinylethyl-1H-pyrrolo[2,3-d]pyrimidine-2,4(3H,7H)-dione derivatives as novel, high-affinity and selective alpha(1)-adrenoceptor ligands. AB - The discovery of a new series of selective and high-affinity alpha(1) adrenoceptor (alpha(1)-AR) ligands, characterized by a 1H-pyrrolo[2,3-d] pyrimidine-2,4(3H,7H)-dione system, is described in this paper. Some synthesized compounds, including 20, 22, and 30, displayed affinity in the nanomolar range for alpha(1)-ARs and substantial selectivity with respect to 5-HT(1A) and dopaminergic D(1) and D(2) receptors. Functional assays, performed on selected derivatives, showed antagonistic properties. PMID- 16216507 TI - Synthesis and evaluation of novel 8,6-fused bicyclic peptidomimetic compounds as interleukin-1beta converting enzyme inhibitors. AB - Two novel 8,6-fused bicyclic peptidomimetic ring systems were synthesized utilizing olefin metathesis as the key reaction for the formation of the eight membered ring. Both peptidomimetic scaffolds were further elaborated into potent ICE inhibitors, with numerous compounds exhibiting caspase-1 IC(50)s less than 10nM. PMID- 16216508 TI - Discovery of novel 1-arylmethyl pyrrolidin-2-yl ethanol amines as calcium-sensing receptor antagonists. AB - A 3D quantitative structure-activity relationship study for inhibition of calcium sensing receptor in the aryloxypropanolamine series predicted that these molecules adopt a U-shaped conformation with pi-stacking between the two aromatic rings. This hypothesis led to the discovery of novel 1-arylmethyl pyrrolidin-2-yl ethanol amines capable of antagonizing the calcium-sensing receptor with potency comparable to that of NPS-2143. PMID- 16216509 TI - Novel triazole based inhibitors of Ras farnesyl transferase. AB - A novel series of potent inhibitors of Ras farnesyl transferase possessing a 1,2,4-triazole pharmacophore is described. These inhibitors were discovered from a parallel synthesis effort and were subsequently optimized to in vitro IC(50) value of less than 1nM. PMID- 16216510 TI - Type II secretion: a protein secretion system for all seasons. AB - In Gram-negative bacteria, type II secretion (T2S) is one of five protein secretion systems that permit the export of proteins from within the bacterial cell to the extracellular milieu and/or into target host cells. An analysis of numerous sequenced genomes now reveals that T2S genes are common, but by no means universal, in Gram-negative bacteria. Recent functional studies indicate that T2S can promote the virulence of human, animal and plant pathogens, as well as the physiology of various environmental bacteria. Thus, it is an opportune time to highlight the new and different ways in which T2S serves bacterial function. PMID- 16216511 TI - Absolute stereochemistry and antitumor activity of iejimalides. AB - The absolute configurations at five chiral centers, except for C-32(S) reported previously, in iejimalides A, C, and D, potent cytotoxic 24-membered macrolides isolated from a tunicate Eudistoma cf. rigida, were assigned as 4R, 9S, 17S, 22S, and 23S on the basis of detailed analysis of NMR data and chemical means. Furthermore, the structures proposed for iejimalides A, C, and D were revised to their 13Z-isomers. Iejimalides A-D (1-4) exhibited antitumor activity in vivo. PMID- 16216512 TI - A chemoenzymatic scalable route to optically active (R)-1-(pyridin-3-yl)-2 aminoethanol, valuable moiety of beta3-adrenergic receptor agonists. AB - Enantiomerically pure (R)-2-chloro-1-(pyridin-3-yl)ethanol has been prepared by kinetic resolution performed in the presence of Candida antarctica SP435-L lipase immobilized on a macroporous polyacrylate resin (Novozym 435). It was converted into (R)-1-(pyridin-3-yl)-2-aminoethanol, left-hand side of beta(3)-adrenergic receptor ligands. PMID- 16216513 TI - 3'-N-Alkylamino-3'-deoxy-ara-uridines: a new class of potential inhibitors of ribonuclease A and angiogenin. AB - In this study, we report the inhibition of ribonuclease A (RNase A) by certain aminonucleosides. This is the first such instance of the use of this group of compounds to investigate the inhibitory activity of this protein. The compounds synthesized have been tested for their ability to inhibit the ribonucleolytic activity of RNase A by an agarose gel-based assay. A tRNA precipitation assay and inhibition kinetic studies with cytidine 2',3'-cyclic monophosphate as the substrate have also been conducted for two of the compounds. Results indicate substantial inhibitory activity with inhibition association constants in the micromolar range. The experimental studies have been substantiated by docking of the aminonucleoside ligands to RNase A using AutoDock. We find that the ligands preferentially bind to the active site of the protein molecule with a favorable free energy of binding. The study has been extended to a member of the ribonuclease superfamily, angiogenin, which is a potent inducer of blood vessel formation. We show that the aminonucleosides act as potent inhibitors of angiogenin induced angiogenesis. PMID- 16216514 TI - Phenazine-1-carboxamides: structure-cytotoxicity relationships for 9-substituents and changes in the H-bonding pattern of the cationic side chain. AB - A series of phenazine-1-carboxamides were prepared, including variations in both chromophore substituents and the nature of the cationic side chain. The novel side-chain analogues were prepared from the corresponding phenazine-1-carboxylic acids via Schmidt conversion to the 1-amines and from the corresponding 1 halides. Structure-cytotoxicity relationships for these compounds in a panel of tumor cell lines showed that there is very limited scope for variation of the structure of the 1-carboxamide side chain, consistent with the recent structural model of how tricyclic carboxamides bind to DNA. There was generally little difference in IC(50)s between parent and P-glycoprotein expressing cell lines, suggesting that most of the compounds are not affected by the presence of this efflux pump. PMID- 16216516 TI - C-Glycoside analogues of beta-galactosylceramide with a simple ceramide substitute: synthesis and binding to HIV-1 gp120. AB - The synthesis and HIV-1 gp120 binding of C- and aza-C-glycoside analogues of beta galactosylceramide (GalCer) that contain a simple C-17 hydrocarbon chain as a ceramide substitute are described. Both compounds originate from stearic acid, and a carbohydrate-derived thioacetal-alcohol, and their syntheses are potentially general for beta-C-galactosides and their aza-C-partners. They showed potent and specific affinity for gp120 in an assay based on the change of surface pressure when the glycolipid monolayers were exposed to solutions of gp120. Interestingly, the aza-C-glycoside exhibited a significantly higher affinity than GalCer, whereas the C-glycoside was as active as GalCer. PMID- 16216515 TI - Potent, selective, and orally bioavailable matrix metalloproteinase-13 inhibitors for the treatment of osteoarthritis. AB - Modification of alpha-biphenylsulfonamidocarboxylic acids led to potent and selective MMP-13 inhibitors. Compound 16 showed 100% oral bioavailability in rats and demonstrated >50% inhibition of bovine cartilage degradation at 10 ng/mL. PMID- 16216517 TI - Putative drug binding conformations of monoamine transporters. AB - Structural information about monoamine transporters and their interactions with psychotropic drugs is important for understanding their molecular mechanisms of action and for drug development. The crystal structure of a Major Facilitator Superfamily (MFS) transporter, the lactose permease symporter (lac permease), has provided insight into the three-dimensional structure and mechanisms of secondary transporters. Based on the hypothesis that the 12 transmembrane alpha-helix (TMH) secondary transporters belong to a common folding class, the lac permease structure was used for molecular modeling of the serotonin transporter (SERT), the dopamine transporter (DAT), and the noradrenaline transporter (NET). The molecular modeling methods used included amino acid sequence alignment, homology modeling, and molecular mechanical energy calculations. The lac permease crystal structure has an inward-facing conformation, and construction of outward-facing SERT, DAT, and NET conformations allowing ligand binding was the most challenging step of the modeling procedure. The psychomotor stimulants cocaine and S amphetamine, and the selective serotonin reuptake inhibitor (SSRI) S-citalopram, were docked into putative binding sites on the transporters to examine their molecular binding mechanisms. In the inward-facing conformation of SERT the translocation pore was closed towards the extracellular side by hydrophobic interactions between the conserved amino acids Phe105, Pro106, Phe117, and Ala372. An unconserved amino acid, Asp499 in TMH10 in NET, may contribute to the low affinity of S-citalopram to NET. PMID- 16216518 TI - Activity-guided isolation of cytotoxic constituents from the bark of Aglaia crassinervia collected in Indonesia. AB - Activity-guided fractionation of a CHCl(3)-soluble partition of the MeOH extract of the bark of Aglaia crassinervia collected in Indonesia led to the isolation of three new glabretal-type triterpenoids, aglaiaglabretols A-C (1-3), as well as nine known compounds, 3-epi-cabraleahydroxylactone (4), cabraleahydroxylactone (5), rocaglaol (6), 2beta,3beta-dihydroxy-5alpha-pregn-17(20)-(E)-16-one, scopoletin, and mixtures of cabraleadiol and epicotillol and of beta-sitosterol and stigmasterol. The structures of compounds 1-3 were determined on the basis of spectroscopic and chemical methods. The structure of aglaiaglabretol A (1) was confirmed by single-crystal X-ray analysis, and the absolute stereochemistry of this isolate was established by the Mosher ester method. The cytotoxic activity of all isolates and several chemical transformation products obtained in the present study was evaluated. The known cyclopenta[b]benzofuran, rocaglaol (6), was found to be significantly active and comparable in potency to the positive controls, paclitaxel and camptothecin. Aglaiaglabretol B (2) was further tested in an in vivo hollow fiber model. PMID- 16216519 TI - Parallel synthesis of 9-aminoacridines and their evaluation against chloroquine resistant Plasmodium falciparum. AB - A parallel synthetic strategy to the 9-aminoacridine scaffold of the classical anti-malarial drug quinacrine (2) is presented. The method features a new route to 9-chloroacridines that utilizes triflates of salicylic acid derivatives, which are commercially available in a variety of substitution patterns. The route allows ready variation of the two diversity elements present in this class of molecules: the tricyclic aromatic heterocyclic core, and the disubstituted diamine sidechain. In this study, a library of 175 compounds was designed, although only 93 of the final products had purities acceptable for screening. Impurity was generally due to incomplete removal of 9-acridones (18), a degradation product of the 9-chloroacridine synthetic intermediates. The library was screened against two strains of Plasmodium falciparum, including a model of the drug-resistant parasite, and six novel compounds were found to have IC(50) values in the low nanomolar range. PMID- 16216520 TI - Bright cyclic light accelerates photoreceptor cell degeneration in tubby mice. AB - Photoreceptor cell death is an irreversible, pathologic event in many blinding retinal diseases including retinitis pigmentosa, age-related macular disease, and retinal detachment. Light exposure can exacerbate a variety of human retinal diseases by increasing the rate of photoreceptor cell death. In the present study, we characterize the kinetics of photoreceptor cell death in Tubby (homozygous tub/tub, which have inherited, progressive retinal degeneration) mice born and raised in a bright cyclic light environment. Our data show that raising tub/tub mice in a bright cyclic light environment induces rapid loss of photoreceptors. This effect can be slowed, but not prevented, by raising animals in constant darkness, which suggests the involvement of phototransduction in the accelerated death of photoreceptors in this animal. We further demonstrated that the activities of cytosolic cytochrome c and caspases-3 and -9 were significantly increased in the retinas of tub/tub mice. Raising animals in darkness significantly reduced the increased activities of caspases-3 and -9, as well as cytosolic cytochrome c. We also observed that rhodopsin, a phototransduction protein, is not restricted to the rod outer segment, but is distributed throughout the rod cell, including the inner segments, cell bodies, and synapses. In addition, the light-dependent translocation and compartmentalization of arrestin and transducin are affected by the tubby mutation. Our results support the interpretation that problems in protein trafficking in the photoreceptors of the tub/tub mouse may contribute to retinal degeneration. PMID- 16216521 TI - Suppression of experimental autoimmune myocarditis by sodium fusidate (fusidin). AB - Fusidic acid and its sodium salt sodium fusidate (fusidin) are widely used antibiotics that possess immunomodulating properties. It has been shown that fusidin ameliorates the course of several organ-specific immunoinflammatory diseases and thus we investigated the effect of fusidin on myosin-induced experimental autoimmune myocarditis (EAM) in rats, a well-established animal model for human giant cell myocarditis and postmyocarditis dilated cardiomyopathy (DCM). Fusidin at doses of 80 mg kg(-1) was administrated i.m. to male Dark Agouti (DA) rats, either from days 0 to 10 (early treatment group), or from days 10 to 20 (late treatment group) after induction of EAM. Efficacy of fusidin treatment was determined on day 21 of EAM development. It was observed that both early and late treatment with fusidin markedly ameliorated clinical, histological and immunohistochemical signs of the disease. The treated rats had significantly decreased incidence of EAM, heart weight and heart weight/body weight ratio (Hw/Bw) compared with untreated animals. In contrast to the severe myocardial damage and cellular infiltration in the EAM rats, there was only focal infiltration of inflammatory cells in the myocardium of the treated rats. In both treated groups the mean microscopic score was markedly lower compared with vehicle-treated animals. In addition, the number of CD4+, ED1+ and OX6+ cells was significantly lower in myocardium of fusidin-treated rats than that in untreated group. The present findings suggest that fusidin exhibited both early and late therapeutical effect in EAM. PMID- 16216523 TI - Piezoelectric biosensors: strategies for coupling nucleic acids to piezoelectric devices. AB - The development of a piezoelectric biosensor based on nucleic acids interaction is presented focusing on the methodology for probe immobilization. This is a key step in any DNA biosensor development. Often, the detection limits and, in general, the analytical performances of the biosensor can be improved by optimizing the immobilization of the receptor on the transducer surface. DNA must be attached to the solid support, retaining native conformation, and binding activity. This attachment must be stable over the course of a binding assay and, in addition, sufficient binding sites must be presented to the solution phase to interact with the analyte. In this paper, the optimization of the coating of the gold quartz crystal surface, to immobilize an oligonucleotide probe, is reported. Two immobilization procedures are illustrated in details with a comparison regarding the immobilization of the probe, the detection of the hybridization reaction, and the possibility of regeneration. The two procedures are based on the use of biotinylated or thiolated DNA probes. Specific applications will be also presented. PMID- 16216524 TI - Expression of glycosylated haloalkane dehalogenase LinB in Pichia pastoris. AB - Heterologous expression of the bacterial enzyme haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26 in methylotrophic yeast Pichia pastoris is reported. The haloalkane dehalogenase gene linB was subcloned into the pPICZalphaA vector and integrated into the genome of P. pastoris. The recombinant LinB secreted from the yeast was purified to homogeneity and biochemically characterized. The deglycosylation experiment and mass spectrometry measurements showed that the recombinant LinB expressed in P. pastoris is glycosylated with a 2.8 kDa size of high mannose core. The specific activity of the glycosylated LinB was 15.6 +/- 3.7 micromol/min/mg of protein with 1,2-dibromoethane and 1.86 +/- 0.36 micromol/min/mg of protein with 1-chlorobutane. Activity and solution structure of the protein produced in P. pastoris is comparable with that of recombinant LinB expressed in Escherichia coli. The melting temperature determined by the circular dichroism (41.7+/-0.3 degrees C for LinB expressed in P. pastoris and 41.8 +/- 0.3 degrees C expressed in E. coli) and thermal stability measured by specific activity to 1-chlorobutane were also similar for two enzymes. Our results show that LinB can be extracellularly expressed in eukaryotic cell and glycosylation had no effect on activity, protein fold and thermal stability of LinB. PMID- 16216525 TI - Expression and purification of a single-chain variable fragment antibody derived from a polyol-responsive monoclonal antibody. AB - A previously described polyol-responsive monoclonal antibody (PR-mAb) was converted to a single-chain variable fragment (scFv). This antibody, PR-mAb NT73, reacts with the beta' subunit of Escherichia coli RNA polymerase and has been used for the immunoaffinity purification of polymerase. mRNAs encoding the variable regions of the heavy chain (VH) and light chain (VL) were used as the template for cDNA synthesis. The sequences were joined by the addition of a "linker" sequence and then cloned into several expression vectors. A variety of expression plasmids and E. coli hosts were used to determine the optimal expression system. Expression was highest with the pET22b(+) vector and the Rosetta(DE3)pLysS host strain, which produced approximately 60 mg purified His tagged scFv per liter of culture (3.3 g wet weight cells). Although the production of soluble scFv was preferred, overproduced scFv formed inclusion bodies under every expression condition. Therefore, inclusion bodies had to be isolated, washed, solubilized, and refolded. The FoldIt protein refolding kit and enzyme-linked immunosorbent assay were sequentially used to determine the optimal refolding conditions that would produce active His-tagged scFv. Immobilized metal affinity chromatography was used for the final purification of the refolded active scFv. The polyol-responsiveness of the scFv was determined by an ELISA elution assay. Although the scFv loses considerable affinity for its antigen, it maintains similar polyol-responsiveness as the parent monoclonal antibody, PR-mAb NT73. PMID- 16216522 TI - An extended vision for dynamic high-resolution intravital immune imaging. AB - The past few years have seen the application of confocal and especially two photon microscopy to the dynamic high-resolution imaging of lymphocytes and antigen presenting cells within organs such as lymph nodes and thymus. After summarizing some of the published results obtained to date using these methods, we describe our view of how this technology will develop and be applied in the near future. This includes its extension to a wide variety of non-lymphoid tissues, to the tracking of functional responses in addition to migratory behavior, to the analysis of molecular events previously studied only in vitro, to dissection of the interplay between hematopoietic and stromal elements, to visualization of a wider array of cell types including neutrophils, macrophages, NK cells, NKT cells and others, and to the interaction of the host with infectious agents. Reaching these goals will depend on a combination of new tools for genetic manipulations, novel fluorescent reporters, enhanced instrumentation, and better surgical techniques for the extended imaging of live animals. The end result will be a new level of understanding of how orchestrated cell movement and interaction contribute to the physiological and pathological activities of the immune system. PMID- 16216526 TI - Fusion expression of bovine lactoferricin in Escherichia coli. AB - The drug resistance problem has been growing with the utilization of current antibiotics in feed and medical industries. LfcinB, a 25-amino acid antibacterial peptide derived from bovine lactoferrin, is one of potential alternatives of antibiotics. According to the bias of codon utilization of Escherichia coli, a fragment encoding LfcinB has been chemically synthesized, inserted into vector pGEX-4T-2 and expressed in E. coli. The antibacterial peptide was fused with GST with a protease cleavage site located between them. Two constructs with different cleavage sites were made. One construct, pGEX-Th-LfcinB, contains a thrombin cleavage site carried by the vector, and the other, pGEX-Th-Xa-LfcinB, contains a Factor Xa cleavage site which was introduced after the thrombin cleavage site. Fusion protein GST-Th-LfcinB protein was efficiently cleaved by thrombin, yielding recombinant LfcinB showing antibacterial activity. However, fusion protein GEX-Th-Xa-Lfcin B containing Factor Xa recognition site could not be cleaved by Factor Xa at the conditions tried in this study. Successful expression of LfcinB in E. coli provides a possible method to produce LfcinB in large amounts. PMID- 16216527 TI - Design and optimization of a recombinant system for large-scale production of the MPT64 antigen from Mycobacterium tuberculosis. AB - Early clinical trials of a potential new tuberculosis (TB) diagnostic, the Patch Test for Active TB (PTAT), used MPB64 protein that was purified from the spent medium of Bacillus Calmette-Guerin (BCG) Tokyo 172 vaccine production. The yield was poor, 0.05 mg/L, and the process for purification of the protein was complex, requiring four chromatographic steps. The combination of yield and purification complexity compromised the ability to produce the PTAT diagnostic in quantities sufficient for larger clinical trials and commercialization. We report here a highly efficient method for the overexpression and purification of recombinant MPT64 from Escherichia coli (rMPT64) based upon a mild insolubility of rMPT64 following induction, and scalable anion-exchange and gel filtration chromatographies. Yields of protein were improved substantially to approximately 250 mg/L, and resulted in a preparation greater than 98% pure. Quantitative release assays were developed and used with MALDI-TOF mass spectrometry to confirm the identity of rMPT64. Using a guinea pig model of active TB, we found that rMPT64 elicited a specific immune response indistinguishable from that of MPB64 purified from BCG Tokyo culture filtrates. These results describe the first efficient and scalable protocol for production of rMPT64, demonstrate its activity in an animal model of active TB, and lay the foundation of ongoing and future use of the PTAT in clinical settings. PMID- 16216528 TI - Characterization of Gly-D-Phe, Gly-L-Leu, and D-Phe as affinity ligands to thermolysin. AB - In this study, glycyl-D-phenylalanine (Gly-D-Phe), glycyl-L-leucine (Gly-L-Leu), and D-phenylalanine (D-Phe) were characterized for their abilities as affinity ligands to thermolysin. Each of the ligands was immobilized to the resin. The optimum pH for adsorption of thermolysin is 5.0-6.0 for each of the ligands. By the affinity column chromatography in which 2mg thermolysin was applied onto 4 ml volume of the resins at pH 5.5, the adsorption ratios based on casein hydrolysis activity were 100% for each of the ligands. However, the adsorption ratios of the resins containing Gly-L-Leu and D-Phe, unlike that of Gly-D-Phe, were progressively decreased with increasing the amounts of thermolysin applied to the column. Measurement of adsorption isotherms showed that the association constant to thermolysin at pH 5.5 of the resins containing Gly-D-Phe was (3.3+/ 0.8)x10(5)M(-1), while those of Gly-L-Leu and D-Phe were approximately ten times less. This result is coincident with the observations of performances in affinity column chromatography. On the other hand, maximum thermolysin binding capacities were almost the same among the resins examined. These results indicate that Gly-D Phe is more suitable than Gly-L-Leu and D-Phe as an affinity ligand for purification of thermolysin. PMID- 16216529 TI - Phenylpropanolamine and hemorrhagic stroke in the Hemorrhagic Stroke Project: a reappraisal in the context of science, the Food and Drug Administration, and the law. AB - The report of the Hemorrhagic Stroke Project (HSP), a case-control study of phenylpropanolamine (PPA) was the primary reason that the US Food and Drug Administration (FDA) requested that PPA-containing products voluntarily be withdrawn from the market. In subsequent litigation, scientific information emerged that was not available during the deliberations of the FDA and its advisory committee. Our reappraisal leads us to conclude that chance, bias, and confounding are plausible alternative explanations for the observed findings. Thus, we believe that it is not possible to conclude that there is any valid statistical association between PPA and hemorrhagic stroke, let alone make any judgment of causality. Our reappraisal suggests the FDA's regulatory request may have been premature. PMID- 16216530 TI - Sex hormone-binding globulin and serum testosterone are inversely associated with C-reactive protein levels in postmenopausal women at high risk for cardiovascular disease. AB - PURPOSE: C-reactive protein (CRP), androgens, and menopausal loss of endogenous estrogens are associated with cardiovascular disease (CVD). We hypothesized that high androgens, low estradiol, and low sex hormone-binding globulin (SHBG) would be associated with high CRP in postmenopausal women. METHODS: CRP, SHBG, estradiol, and total testosterone were measured using baseline bloods of 221 hormone therapy (HT)-nonusers and 162 HT-users from a cross-sectional analysis in a nested case-control sample of the Women's Health Study. Hormones and CRP were ln-transformed and relationships were assessed with Spearman correlations and linear regression. RESULTS: ln-SHBG (beta=-0.40; p<0.01) and ln-testosterone (beta=-0.24; p=0.04) were the only independent hormonal predictors of ln-CRP among HT-nonusers after adjusting for age, hypertension, smoking, body mass index, diabetes, exercise, HDL cholesterol, alcohol intake, and CVD occurrence during follow-up. Upon stratification, the association between ln-SHBG and ln-CRP persisted among HT nonusers who subsequently developed CVD (beta=-0.55; p=0.01), but not among women who remained CVD-free (p=0.28). The inverse relationship between ln-SHBG and ln-CRP was strongest among the leanest women. None of the sex hormones predicted ln-CRP among HT-users. CONCLUSIONS: SHBG and total testosterone were inversely associated with CRP among HT nonusers in this study. The relationship between SHBG and CRP was more strongly inverse among leaner women. PMID- 16216531 TI - Parametric modulation of cortical activation during smooth pursuit with and without target blanking. an fMRI study. AB - Smooth pursuit eye movements (SPEM) are performed to track slowly moving visual targets and are accompanied by saccades whenever foveal representation is lost. In the present study, we correlated the cerebral activation as assessed by functional magnetic resonance imaging with parameters of eye movement performance in order to determine the cortical areas involved in the retinal and extraretinal processing of maintaining smooth pursuit velocity (SPV) and generating saccades in 16 healthy males. The stimulus consisted of a target moving at a constant velocity of 10 degrees/s with and without target blanking. During constant target presentation, SPV was positively correlated with the BOLD signal in the right V5 complex and negatively correlated with the BOLD response in the left dorsolateral prefrontal cortex (DLPFC). In the condition with target blanking, additional negative correlations with SPV were found in the left frontal eye field (FEF), the left parietoinsular vestibular cortex (PIVC) and the left angular gyrus. Saccadic frequency was negatively correlated with activations of the right mesial intraparietal sulcus (IPS) during both conditions and the right premotor area during continuous target presentation. We conclude that V5 is directly related to the maintenance of an optimal smooth pursuit velocity during visual feedback, whereas the FEF, PFC, angular gyrus and PIVC are involved in reconstitution and prediction whenever SPV decreases, especially during maintenance of smooth pursuit in the absence of a visual target. Furthermore, we suggest that parietal areas are related to the suppression of saccades during smooth pursuit. PMID- 16216532 TI - MAP-based kinetic analysis for voxel-by-voxel compartment model estimation: detailed imaging of the cerebral glucose metabolism using FDG. AB - We propose a novel algorithm for voxel-by-voxel compartment model analysis based on a maximum a posteriori (MAP) algorithm. Voxel-by-voxel compartment model analysis can derive functional images of living tissues, but it suffers from high noise statistics in voxel-based PET data and extended calculation times. We initially set up a feature space of the target radiopharmaceutical composed of a measured plasma time activity curve and a set of compartment model parameters, and measured the noise distribution of the PET data. The dynamic PET data were projected onto the feature space, and then clustered using the Mahalanobis distance. Our method was validated using simulation studies, and compared with ROI-based ordinary kinetic analysis for FDG. The parametric images exhibited an acceptable linear relation with the simulations and the ROI-based results, and the calculation time took about 10 min. We therefore concluded that our proposed MAP-based algorithm is practical. PMID- 16216533 TI - Localizing human visual gamma-band activity in frequency, time and space. AB - Neuronal gamma-band (30-100 Hz) synchronization subserves fundamental functions in neuronal processing. However, different experimental approaches differ widely in their success in finding gamma-band activity. We aimed at linking animal and human studies of gamma-band activity and at preparing optimized methods for an in depth investigation of the mechanisms and functions of gamma-band activity and gamma-band coherence in humans. In the first step described here, we maximized the signal-to-noise ratio with which we can observe visually induced gamma-band activity in human magnetoencephalographic recordings. We used a stimulus and task design that evoked strong gamma-band activity in animals and combined it with multi-taper methods for spectral analysis and adaptive spatial filtering for source analysis. With this approach, we found human visual gamma-band activity very reliably across subjects and across multiple recording sessions of a given subject. While increases in gamma-band activity are typically accompanied by decreases in alpha- and beta-band activity, the gamma-band enhancement was often the spectral component with the highest signal-to-noise ratio. Furthermore, some subjects demonstrated two clearly separate visually induced gamma bands, one around 40 Hz and another between 70 and 80 Hz. Gamma-band activity was sustained for the entire stimulation period, which was up to 3 s. The sources of gamma-band activity were in the calcarine sulcus in all subjects. The results localize human visual gamma-band activity in frequency, time and space and the described methods allow its further investigation with great sensitivity. PMID- 16216534 TI - Trauma modulates amygdala and medial prefrontal responses to consciously attended fear. AB - Effective fear processing relies on the amygdala and medial prefrontal cortex (MPFC). Post-trauma reactions provide a compelling model for examining how the heightened experience of fear impacts these systems. Post-traumatic stress disorder (PTSD) has been associated with excessive amygdala and a lack of MPFC activity in response to nonconscious facial signals of fear, but responses to consciously processed facial fear stimuli have not been examined. We used functional MRI to elucidate the effect of trauma reactions on amygdala-MPFC function during an overt fear perception task. Subjects with PTSD (n = 13) and matched non-traumatized healthy subjects (n = 13) viewed 15 blocks of eight fearful face stimuli alternating pseudorandomly with 15 blocks of neutral faces (stimulus duration 500 ms; ISI 767 ms). We used random effects analyses in SPM2 to examine within- and between-group differences in the MPFC and amygdala search regions of interest. Time series data were used to examine amygdala-MPFC associations and changes across the first (Early) versus second (Late) phases of the experiment. Relative to non-traumatized subjects, PTSD subjects showed a marked bilateral reduction in MPFC activity (in particular, right anterior cingulate cortex, ACC), which showed a different Early-Late pattern to non traumatized subjects and was more pronounced with greater trauma impact and symptomatology. PTSD subjects also showed a small but significant enhancement in left amygdala activity, most apparent during the Late phase, but reduction in Early right amygdala response. Over the time course, trauma was related to a distinct pattern of ACC and amygdala connections. The findings suggest that major life trauma may disrupt the normal pattern of medial prefrontal and amygdala regulation. PMID- 16216536 TI - Model type, implicit data weighting, and model averaging in phylogenetics. PMID- 16216535 TI - Mapping cerebral blood flow changes during auditory-cued conditioned fear in the nontethered, nonrestrained rat. AB - Conditioned fear (CF) is one of the most frequently used behavioral paradigms; however, little work has mapped changes in cerebral perfusion during CF in the rat-the species which has dominated CF research. Adult rats carrying an implanted minipump were exposed to a tone (controls, n = 8) or a tone conditioned in association with footshocks (CS group, n = 9). During reexposure to the tone 24 h later, animals were injected intravenously by remote activation with [14C] iodoantipyrine using the pump. Significant group differences in regional CBF related tissue radioactivity (CBF-TR) were determined by region-of-interest analysis of brain autoradiographs, as well as in the reconstructed, three dimensional brain by statistical parametric mapping (SPM). CS animals demonstrated significantly greater, fear-enhanced increases in CBF-TR in auditory cortex than controls. The lateral amygdala was activated, whereas the basolateral/basomedial and central amygdala were deactivated. In the hippocampus and medial prefrontal cortex, CBF-TR increased significantly ventrally but not dorsally. Significant activations were noted in medial striatum and the thalamic midline and intralaminar nuclei. However, the ventrolateral/dorsolateral striatum and its afferents from motor and somatosensory cortex were deactivated, consistent with the behavioral immobility seen during CF. Significant activations were also noted in the lateral septum, periaqueductal gray, and deep mesencephalic nucleus/tegmental tract. Our results show that auditory stimuli endowed with aversive properties through conditioning result in significant redistribution of cerebral perfusion. SPM is a useful tool in the brain mapping of complex rodent behaviors, in particular the changes in activation patterns in limbic, thalamic, motor, and cortical circuits during CF. PMID- 16216538 TI - Investigations of the induction of the goat beta(C) globin gene by erythropoietin: studies in transgenic mice. AB - The genes of the beta globin locus of sheep and goats undergo three developmental switches, from embryonic epsilon to fetal gamma to juvenile beta(C) and from beta(C) to adult beta(A). The juvenile beta(C) gene of adult goats and sheep are strikingly induced by erythropoietin (Epo). To obtain insights on the mechanism of beta(C) induction by Epo, we produced transgenic mice carrying various beta(C) gene constructs and examined the inducibility of the beta(C) gene following administration of high doses of erythropoietin. None of the treatments resulted in reproducible induction of the beta(C) gene by erythropoietin. We conclude that the Epo inducibility elements are not contained in the 4.7 kb (which included 0.88 kb of the beta(C) promoter and 2.3 kb downstream sequence) beta(C) gene we used or that a trans-acting environment specific to the goat and sheep erythroid cell lineage is required for induction of the beta(C) globin gene by erythropoietin. PMID- 16216539 TI - Application of the chirp z-transform to MRI data. AB - A version of the chirp z-transform (CZT) enabling signal intensity and phase preserving field-of-view scaling has been programmed. The algorithm is important for all single-point imaging sequences such as SPRITE when used with multiple data acquisition for T2* mapping or signal averaging. CZT has particular utility for SPRITE imaging of nuclei with short relaxation times such as sodium at high field. Here, a complete theory of the properties of CZT is given. This method operates entirely in k-space. It is compared with a conventional interpolation approach that works in image space after the application of a fast Fourier transformation. PMID- 16216540 TI - Spatially resolved measurement of rock core porosity. AB - Density weighted, centric scan, Conical SPRITE MRI techniques are applied in the current work for local porosity measurements in fluid saturated porous media. The methodology is tested on a series of sandstone core samples. These samples vary in both porosity and degree of local heterogeneity due to bedding plane structure. The MRI porosity measurement is in good agreement with traditional gravimetric measurements of porosity. Spatially resolved porosity measurements reveal significant porosity variation in some samples. This novel MRI technique should have applications to the characterization of local porosity in a wide variety of porous media. PMID- 16216537 TI - Signature of an early genetic bottleneck in a population of Moroccan sardines (Sardina pilchardus). AB - Fishery assessment models meant to determine sustainability of commercial marine fish failed to predict recent stock collapses due to overexploitation. One flaw of assessment models is that they strongly rely on catch and age-composition statistics, but largely ignore the genetic background of the studied populations. We examined population genetic structure of sardine (Sardina pilchardus) in the centraleastern and northeastern Atlantic Ocean and Mediterranean Sea to aid fishery management of this heavily fished small pelagic species. We found that sardine has a striking mitochondrial control region, and sequenced a fragment of 387 bp of its 5'-end in 261 individuals collected off the coasts of Morocco (Dakhla, Tantan, Safi, Larache, and Nador), Portugal (Quarteira), Spain (Pasajes, Barcelona), and Greece (Kavala). High levels of haplotypic diversity rendered a rather unresolved NJ phylogeny. The recovered tree had no phylogeographic structuring except for the clustering of 13 individuals of Safi. In contrast, individuals grouped together according to the presence or absence of a 13-bp insertion in the sequence. Phi(ST) pairwise comparisons and molecular variance analyses supported genetic differentiation between the population of Pasajes (Bay of Biscay), and those of the Mediterranean Sea and Moroccan coast, with a contact zone around the Strait of Gibraltar. This result confirms the existence of two subspecies, S. pilchardus pilchardus and S. pilchardus sardina that were previously identified based on meristics and morphometry. Mismatch distribution analysis showed that sardine populations are expanding since the Pleistocene. Surprisingly, the population of Safi showed strong and statistically significant levels of genetic differentiation that could be related with isolation and genetic drift. Comparative analysis of the Safi population versus the rest including mismatch distributions, and a Bayesian skyline plot suggest that the Safi population likely underwent an early genetic bottleneck. The genetic singularity of the Safi population could have been responsible for the historical collapse of this sardine stock in the 1970s. PMID- 16216541 TI - Islet neogenesis: a potential therapeutic tool in type 1 diabetes. AB - Current therapies for type 1 diabetes, including fastidious blood glucose monitoring and multiple daily insulin injections, are not sufficient to prevent complications of the disease. Though pancreas and possibly islet transplantation can prevent the progression of complications, the scarcity of donor organs limits widespread application of these approaches. Understanding the mechanisms of beta cell mass expansion as well as the means to exploit these pathways has enabled researchers to develop new strategies to expand and maintain islet cell mass. Potential new therapeutic avenues include ex vivo islet expansion and improved viability of islets prior to implantation, as well as the endogenous expansion of beta-cell mass within the diabetic patient. Islet neogenesis, through stem cell activation and/or transdifferentiation of mature fully differentiated cells, has been proposed as a means of beta-cell mass expansion. Finally, any successful new therapy for type 1 diabetes via beta-cell mass expansion will require prevention of beta-cell death and maintenance of long-term endocrine function. PMID- 16216542 TI - Complex quality traits: now time to model. AB - A recently published dynamic peach model by Francoise Lescourret and Michel Genard simulates changes in quality during final stages of fruit growth. It predicts quality, emerging from complex processes and influenced by environment and management, remarkably well and describes complex emerging behaviour. The model could prove an excellent tool to integrate novel insight from metabolic profiling and pathways of taste-related compounds, ultimately enabling the analysis of gene networks responsible for fruit maturation processes. PMID- 16216543 TI - Chromium picolinate does not produce chromosome damage in CHO cells. AB - Chromium picolinate (CrPic, Chromax) is a dietary supplement that has been commercially available for the past two decades. CrPic has potential benefits for reducing insulin dependence in diabetics by increasing sensitivity of insulin receptors and in stimulating insulin binding. In this study, CrPic was tested for its ability to produce chromosomal aberrations in vitro using Chinese hamster ovary K1 (CHO) cells. CHO cells were exposed to a range of cytotoxic to non cytotoxic concentrations of CrPic for 4 or 20h in the absence of metabolic (S9) activation or for 4h in the presence of S9 activation. CrPic was solubilized with dimethyl sulfoxide (DMSO) to attain the highest possible solubility for maximizing the test doses. Cells were treated with 96.25, 192.5, 385 or 770 microg/mL of CrPic for 4 h in the presence of S9 activation, and for 4 or 20 h in the absence of S9 activation. A distinct precipitate of CrPic was evident in the cell culture medium at 770 microg/mL, which was the highest dose tested. Results showed no statistically significant increases in structural or numerical chromosome aberrations were produced at any test dose level with CrPic in 4-h treatments up to a precipitating dose of 770 microg/mL in either the presence or absence of S9 activation. Additionally no aberrations were observed up to 385 microg/mL (the maximum analyzable dose) following treatment for 20 h in the absence of S9 activation. The percentage of cells with structural or numerical aberrations in CrPic treated cultures was not statistically different (p>0.05) from that quantified in controls at any dose level. The absence of significant differences from control levels demonstrates that CrPic did not induce structural or numerical chromosome aberrations up to doses that were insoluble in the culture medium. PMID- 16216544 TI - Determination of the period for establishment of a liver network echogenic pattern in Schistosoma japonicum infection. AB - Schistosomiasis is caused by infection with Schistosoma haematobium, S. mansoni, S. japonicum, or S. mekongi. S. japonicum infection results in liver cirrhosis at the final stage. A "network" (NW) echogenic pattern on hepatic ultrasonography appears to be specific to S. japonicum infection. The principal aim of the present study was to determine the exact year(s) or even month(s) required for the establishment of the liver NW echogenic pattern from the initial infection in young patients with schistosomiasis japonica since there are few data on this important point. We conducted yearly ultrasonographic, serologic, coprologic, and physical examinations of schistosomiasis patients in the Philippines from 1996 up to the present. During that period, the total number of patients examined was approximately 2,000, among whom we selected 2 patients for determination of the duration required for NW establishment, when they were 10 years old. Although the exact time of initial exposure to schistosomes cannot be determined, the duration for the establishment of NW was definitively confirmed in patient no. 1 to be between 19-24 months based on the results of serologic and coprologic examinations. For patient no. 2, the circumstantial evidence suggested that the establishment of a NW might require 5 to 6 years at maximum. To the best of our knowledge, this is the first evidence-based report on the determination of the period required for the establishment of a liver NW echogenic pattern in S. japonicum infection in the Philippines. PMID- 16216545 TI - Diagnosis of NASH using delayed parenchymal imaging of contrast ultrasound. AB - BACKGROUND AND AIMS: Non alcoholic steatohepatitis (NASH) is one of the representative liver diseases in the developed countries. Diagnosis of NASH is dependent on histological findings from liver biopsy. Usefulness of contrast ultrasound with Levovist for diagnosis of NASH is described. METHODS AND MATERIALS: Clinical study: Ultrasound contrast agent, Levovist of 2.5g was injected intravenously. The liver was scanned at 5, 10, 15, 20, 30, 40, and 50min after Levovist injection in different planes using a contrast specific ultrasound mode. Changes in microbubble accumulation in the liver were evaluated. The signal intensity from regions of interest (ROI) on the contrast images was measured and accumulation and decrescence of microbubbles were estimated using the time intensity curves (TICs). The image data and TICs were evaluated by blind reviewers. Fifteen patients with NASH, 8 with alcoholic steatohepatitis (ASH), 45 with non alcoholic fatty liver (NAFL), 10 with chronic hepatitis C (CHC) and 10 healthy volunteers were studied. Animal study: Methionine-choline-deficient diet (MCDD) fed rats were used for NASH model. Correlation between microbubble accumulation and morphological and functional changes of sinusoidal endothelium and macrophage was evaluated. RESULTS: The maximum intensity of contrast ultrasound was decreased and time course decrescence was more rapid in NASH than the other groups. These changes were correlated to the degree of centrilobular and pericellular fibrosis but not to steatosis in histological study. Disturbance of microbubble accumulation was correlated with sinusoidal function rather than morphological changes such as fibrosis and parenchymatitis in the animal studies. CONCLUSIONS: The Levovist contrast study enables differential diagnosis between NASH and other diseases that provoke steatosis and fibrosis. PMID- 16216546 TI - Obesity and NASH in Japan. AB - BACKGROUND: NASH is expected to be a major target disease in near future, although we are too short in information to realize NASH. In this study, we revealed the prevalence of fatty liver, which is a background disease of NASH, in Kyoto. Then we made histological diagnosis of NASH from chronic liver disorder with fatty liver. MATERIALS AND METHODS: By using the data obtained from medical check-up with more than 1000 subjects per year in Kyoto, the prevalence of fatty liver on ultrasound, serum ALT, body mass index (BMI) were compared between 1994 and 2004. Next 32 patients with elevated ALT and bright liver on ultrasound were biopsied in order to diagnose them as NASH or not. Biochemical, serological and hormonal parameters as well as adipocytokine profiles were also studied. Lastly, liver tissues from those patients were examined to compare the expression of peroxisome proliferator activator receptor (PPAR) alpha and gamma. RESULTS: It was revealed that the prevalence of fatty liver increased from 12.9 to 34.7% over 10 years. The mean ALT and BMI also increased in this period. From 32 patients, 6 cases were excluded for alcohol, autoimmune hepatitis, and drug-induced liver injury, leaving 26 cases to enter the study. Among them, 12 and 8 patients showed grade 1 and grade 2 steatohepatitis, which were consistant with NASH. Various data were compared among simple steatosis, grade 1 steatohepatitis, and grade 2 steatohepatitis, but highly sensitive CRP was the sole parameter with statistical significance. PPAR alpha seems to be expressed higher in patients with steatohepatitis, although PPAR alpha did not show any significance. CONCLUSION: We found the prevalence of fatty liver was increasing over 10 years. The histological examination showed more patients with grade 1 or grade 2 steatohepatitis than expected, which means we might have much more NASH cases undiagnosed. We were unable to propose reliable diagnostic markers to diagnose NASH from this study. PMID- 16216547 TI - Polyunsaturated fatty acids inhibit telomerase activity in DLD-1 human colorectal adenocarcinoma cells: a dual mechanism approach. AB - As high telomerase activity is detected in most cancer cells, telomerase represents a promising cancer therapeutic target. We investigated the inhibitory effect of various fatty acids on telomerase, with particular emphasis on those with antitumor properties, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). To evaluate the direct effect of fatty acids on telomerase, cell lysates of DLD-1 human colorectal adenocarcinoma cells were mixed with sample fatty acids, and the telomerase activity was determined. Saturated fatty acids and trans-fatty acids showed very weak or no inhibition of telomerase. In contrast, cis-unsaturated fatty acids significantly inhibited the enzyme, and the inhibitory potency was elevated with an increase in the number of double bonds. Accordingly, polyunsaturated fatty acids (PUFAs), like EPA and DHA, appeared to be powerful telomerase inhibitors. To assess the transcriptional effect, DLD-1 cells were cultured in the presence of sample fatty acids, and telomerase activity and gene expression were subsequently evaluated. Culturing DLD-1 cells with either EPA or DHA resulted in a remarkable decrease in telomerase activity. EPA and DHA inhibited telomerase by down-regulating human telomerase reverse transcriptase (hTERT) and c-myc expression via protein kinase C inhibition. These results indicate that PUFAs can directly inhibit the enzymatic activity of telomerase as well as modulate the telomerase at the transcriptional level. PMID- 16216548 TI - Dietary coconut oil increases conjugated linoleic acid-induced body fat loss in mice independent of essential fatty acid deficiency. AB - Conjugated linoleic acid (CLA) induces a body fat loss that is enhanced in mice fed coconut oil (CO), which lacks essential fatty acids (EFA). Our objective was to determine if CO enhancement of CLA-induced body fat loss is due to the lack of EFA. The CLA-EFA interaction was tested by feeding CO and fat free (FF) diets for varying times with and without replenishment of individual EFA. Mice fed CO during only the 2-week CLA-feeding period did not differ from control mice in their adipose EFA content but still tended (P=0.06) to be leaner than mice fed soy oil (SO). Mice raised on CO or FF diets and fed CLA were leaner than the SO+CLA-fed mice (P<0.01). Mice raised on CO and then replenished with linoleic, linolenic, or arachidonic acid were leaner when fed CLA than mice raised on SO (P<0.001). Body fat of CO+CLA-fed mice was not affected by EFA addition. In summary, CO-fed mice not lacking in tissue EFA responded more to CLA than SO-fed mice. Also, EFA addition to CO diets did not alter the enhanced response to CLA. Therefore, the increased response to CLA in mice raised on CO or FF diets appears to be independent of a dietary EFA deficiency. PMID- 16216549 TI - Pulmonary surfactant function is abolished by an elevated proportion of cholesterol. AB - A molecular film of pulmonary surfactant strongly reduces the surface tension of the lung epithelium-air interface. Human pulmonary surfactant contains 5-10% cholesterol by mass, among other lipids and surfactant specific proteins. An elevated proportion of cholesterol is found in surfactant, recovered from acutely injured lungs (ALI). The functional role of cholesterol in pulmonary surfactant has remained controversial. Cholesterol is excluded from most pulmonary surfactant replacement formulations, used clinically to treat conditions of surfactant deficiency. This is because cholesterol has been shown in vitro to impair the surface activity of surfactant even at a physiological level. In the current study, the functional role of cholesterol has been re-evaluated using an improved method of evaluating surface activity in vitro, the captive bubble surfactometer (CBS). Cholesterol was added to one of the clinically used therapeutic surfactants, BLES, a bovine lipid extract surfactant, and the surface activity evaluated, including the adsorption rate of the substance to the air water interface, its ability to produce a surface tension close to zero and the area compression needed to obtain that low surface tension. No differences in the surface activity were found for BLES samples containing either none, 5 or 10% cholesterol by mass with respect to the minimal surface tension. Our findings therefore suggest that the earlier-described deleterious effects of physiological amounts of cholesterol are related to the experimental methodology. However, at 20%, cholesterol effectively abolished surfactant function and a surface tension below 15 mN/m was not obtained. Inhibition of surface activity by cholesterol may therefore partially or fully explain the impaired lung function in the case of ALI. We discuss a molecular mechanism that could explain why cholesterol does not prevent low surface tension of surfactant films at physiological levels but abolishes surfactant function at higher levels. PMID- 16216550 TI - Serine palmitoyl-CoA transferase (SPT) deficiency and sphingolipid levels in mice. AB - Sphingolipids play a very important role in cell membrane formation, signal transduction, and plasma lipoprotein metabolism, and all these functions may have an impact on atherosclerotic development. Serine palmitoyl-CoA transferase (SPT) is the key enzyme in sphingolipid biosynthesis. To evaluate in vivo SPT activity and its role in sphingolipid metabolism, we applied homologous recombination to embryonic stem cells, producing mice with long chain base 1 (Sptlc1) and long chain base 2 (Sptlc2), two subunits of SPT, gene deficiency. Homozygous Sptlc11 and Sptlc2 mice are embryonic lethal, whereas heterozygous versions of both animals (Sptlc1(+/-), Sptlc2(+/-)) are healthy. Analysis showed that, compared with WT mice, Sptlc1(+/-) and Sptlc2(+/-) mice had: (1) decreased liver Sptlc1 and Sptlc2 mRNA by 44% and 57% (P<0.01 and P<0.0001, respectively); (2) decreased liver Sptlc1 mass by 50% and Sptlc2 mass by 70% (P<0.01 and P<0.01, respectively), moreover, Sptlc1 mass decreased by 70% in Sptlc2(+/-) mouse liver, while Sptlc2 mass decreased by 53% in Sptlc1(+/-) mouse liver (P<0.001 and P<0.01, respectively); (3) decreased liver SPT activity by 45% and 60% (P<0.01, respectively); (4) decreased liver ceramide (22% and 39%, P<0.05 and P<0.01, respectively) and sphingosine levels (22% and 31%, P<0.05 and P<0.01, respectively); (5) decreased plasma ceramide (45% and 39%, P<0.01, respectively), sphingosine-1-phosphate (31% and 32%, P<0.01, respectively) and sphingosine levels (22.5% and 25%, P<0.01, respectively); (6) dramatically decreased plasma lysosphingomyelin (17-fold and 16-fold, P<0.0001, respectively); and (7) no change of plasma sphingomyelin, triglyceride, total cholesterol, phospholipids, and liver sphingomyelin levels. These results indicated that both Sptlc1 and Sptlc2 interactions are necessary for SPT activity in vivo, and that SPT activity directly influences plasma sphingolipid levels. Furthermore, manipulation of SPT activity might well influence the course of such diseases as atherosclerosis. PMID- 16216557 TI - Molecular medicine: a lifetime of learning, teaching and caring. PMID- 16216558 TI - BMP7 signaling in renal development and disease. AB - Fibrosis, and in particular tubulointerstitial fibrosis, is a common feature of almost all chronic renal diseases. Over the past several years, significant progress has been made in defining the underlying mechanisms of tubulointerstitial fibrosis. In a variety of mouse models, expression of transforming growth factor-beta is a primary causative factor which leads to increased numbers of myofibroblasts, collagen deposition and loss of tubular epithelia. More recently, another member of the transforming growth factor-beta superfamily, BMP7, was shown to counteract transforming growth factor-beta mediated fibrosis. The activities of these secreted factors are regulated, in part, by extracellular ligand binding proteins which can enhance or suppress receptor ligand interactions. PMID- 16216559 TI - Heterogeneity of heart failure management programs in Australia. AB - BACKGROUND: Heart Failure Management Programs (HFMPs) have proven to be cost effective in minimising recurrent hospitalisations, morbidity and mortality. However, variability between the programs exists which could translate into variable health outcomes. OBJECTIVE: To survey the characteristics of HFMPs throughout Australia and to identify potential heterogeneity in their organisation and structure. METHOD: Thirty-nine post-discharge HFMPs were identified from a systematic search of the Australian health-care system in 2002. A comprehensive 19-item questionnaire specifically examining characteristics of HFMPs was sent to co-ordinators of identified programs in early 2003. RESULTS: All participants responded with six institutions (15%) indicating that their HFMP had ceased operations due to a lack of funding. The survey revealed an uneven distribution of the 33 active HFMPs operating throughout Australia. Overall, 4450 post-discharge HF patients (median: 74; IQR: 24-147) were managed via these programs, representing only 11% of the potential caseload for an Australia-wide network of HFMPs. Heterogeneity of these programs existed in respect to the model of care applied within the program (70% applied a home-based program and 18% a specialist HF clinic) and applied interventions (30% of programs had no discharge criteria and 45% of programs prevented nurses administering/titrating medications). Sustained funding was available to only 52% of the active HFMPs. CONCLUSION: Inequity of access to HFMPs in Australia is evident in relation to locality and high service demand, further complicated by inadequate funding. Heterogeneity between these programs is substantial. The development of national benchmarks for evidence-based HFMPs is required to address program variability and funding issues to realise their potential to improve health outcomes. PMID- 16216560 TI - In vivo genetic selection of renal proximal tubules. AB - Repopulation by transplanted cells can result in effective therapy for several regenerative organs including blood, liver, and skin. In contrast, cell therapies for renal diseases are not currently available. Here we developed an animal model in which cells genetically resistant to a toxic intermediate of tyrosine metabolism, homogentisic acid (HGA), were able to repopulate the damaged proximal tubule epithelium of mice with fumarylacetoacetate hydrolase (Fah) deficiency. HGA resistance was achieved by two independent mechanisms. First, Fah+ transplanted bone marrow cells produced significant replacement of damaged proximal tubular epithelium (up to 50%). The majority of bone marrow-derived epithelial cells were generated by cell fusion, not transdifferentiation. In addition to regeneration by fusion-derived epithelial cells, proximal tubular repopulation was also observed by host epithelial cells, which had lost the homogentisic acid dioxygenase gene. These data demonstrate that extensive regeneration of the renal proximal tubule compartment can be achieved through genetic selection of functional cells. PMID- 16216561 TI - Assaying all of the nitrogen oxides in breath modifies the interpretation of exhaled nitric oxide. AB - Exhaled nitric oxide (NO) assays measure the quantity of NO that emanates from the airway, not the amount of NO that is formed. Consumptive processes-including oxidation reactions-decrease the amount of gas phase NO available for exhalation. Higher oxides of nitrogen (HiNO(x)) are resulting reaction products, and are easily measured in exhaled breath condensate (EBC). We performed concurrent sampling of exhaled breath for gas phase NO and EBC HiNO(x) in controls and stable asthmatics. We identified that, mole for mole, asthma patients hourly exhale more HiNO(x) than they do NO, with a HiNO(x)/NO ratio of 1.21 (0.54-3.4). This is the reverse of the ratio found in controls, in whom the HiNO(x)/NO ratio was 0.75 (0.44-0.93), p=0.04. The sum of the hourly molar exhalation of NO and HiNO(x) was significantly higher in asthmatics (333 nmol/h (221-543) than controls (179 (138-231), p<0.001). We conclude that exhaled oxides of nitrogen are more informative when measured together as opposed to in isolation. We suggest that inflammation can be better evaluated with HiNO(x) and NO measured concurrently, and that the level of oxidation in the lung can be evaluated by comparing the easily measured ratios of HiNO(x) to NO in the exhaled breath. PMID- 16216562 TI - Physical and functional interaction of human nuclear uracil-DNA glycosylase with proliferating cell nuclear antigen. AB - Uracil residues arise in DNA by the misincorporation of dUMP in place of dTMP during DNA replication or by the deamination of cytosine in DNA. Uracil-DNA glycosylase initiates DNA base excision repair of uracil residues by catalyzing the hydrolysis of the N-glycosylic bond linking the uracil base to deoxyribose. In human cells, the nuclear form of uracil-DNA glycosylase (UNG2) contains a conserved PCNA-binding motif located at the N-terminus that has been implicated experimentally in binding PCNA. Here we use purified preparations of UNG2 and PCNA to demonstrate that UNG2 physically associates with PCNA. UNG2 co-eluted with PCNA during size exclusion chromatography and bound to a PCNA affinity column. Association of UNG2 with PCNA was abolished by the addition of 100 mM NaCl, and significantly decreased in the presence of 10 mM MgCl(2). The functional significance of the UNG2.PCNA association was demonstrated by UNG2 activity assays. Addition of PCNA (30-810 pmol) to standard uracil-DNA glycosylase reactions containing linear [uracil-(3)H]DNA stimulated UNG2 catalytic activity up to 2.6-fold. UNG2 activity was also stimulated by 7.5 mM MgCl(2). The stimulatory effect of PCNA was increased by the addition of MgCl(2); however, the dependence on PCNA concentration was the same, indicating that the effects of MgCl(2) and PCNA on UNG2 activity occurred by independent mechanisms. Loading of PCNA onto the DNA substrate was required for stimulation, as the activity of UNG2 on circular DNA substrates was not affected by the addition of PCNA. Addition of replication factor C and ATP to reactions containing 90 pmol of PCNA resulted in two-fold stimulation of UNG2 activity on circular DNA. PMID- 16216563 TI - Identification of kinetin and kinetin riboside in coconut (Cocos nucifera L.) water using a combined approach of liquid chromatography-tandem mass spectrometry, high performance liquid chromatography and capillary electrophoresis. AB - Kinetin (free base and riboside), which was assumed by many scientists to be a synthetic cytokinin plant growth hormone, has been detected for the first time in the endosperm liquid of fresh young coconut fruits ("coconut water"). To facilitate the study, we developed a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the identification and quantification of kinetin and kinetin riboside in purified coconut water extract sample. Following a solid-phase extraction of cytokinins in coconut water using C18 columns, the samples were further purified by Oasis MCX columns and analyzed by LC-MS/MS for kinetin and kinetin riboside. Detection by mass spectrometry was carried out using selected reaction monitoring (SRM) mode, by identifying the putative kinetin and kinetin riboside based on their characteristic fragments. Based on a signal-to-noise ratio of 3, the limits of detection in SRM mode were 0.02 microM and 0.005 microM for kinetin and kinetin riboside, respectively. Furthermore, optimal conditions for a baseline chromatographic separation of 18 cytokinin standards by high performance liquid chromatography (HPLC) were developed. The HPLC method had been employed for the confirmation and further fractionation of kinetin in coconut water extracts. The confirmation and fractionation of kinetin riboside was carried out using a further modified HPLC program due to the presence of other interfering material(s) in the sample matrix. Finally, fractions of putative kinetin and kinetin riboside collected from HPLC eluate of coconut water sample were further authenticated by independent capillary zone electrophoresis (CZE) experiment. PMID- 16216565 TI - Risk scoring in acute upper gastrointestinal haemorrhage. PMID- 16216564 TI - Baker's yeast as a tool for the development of antifungal kinase inhibitors- targeting protein kinase C and the cell integrity pathway. AB - Today, the yeast Saccharomyces cerevisiae is probably the best-studied eukaryotic organism. This review first focuses on the signaling process which is mediated by the unique yeast protein kinase C (Pkc1p) and a downstream mitogen-activated protein kinase (MAPK) cascade. This pathway ensures cellular integrity by sensing cell surface stress and controlling cell wall biosynthesis and progression through the cell cycle. The domain structure of Pkc1p is conserved from yeast to humans. A yeast system for heterologous expression of specific domains in a chimeric yeast/mammalian PKC enzyme ("domain shuffling") is depicted. It is also proposed how this system could be employed for the study of protein kinase inhibitors in high-throughput screens. Moreover, a reporter assay that allows a quantitative readout of the activity of the cell integrity signaling pathway is introduced. Since a variety of protein kinases take part in the signal transduction, this broadens the range of targets for potential inhibitors. PMID- 16216566 TI - Endoscopic follow-up after colorectal cancer resection: an Italian multicentre study. AB - BACKGROUND: Endoscopic follow-up is advised in patients operated for colorectal cancer due to a high risk for both metachronous colorectal cancer and adenomas. Such issue has been scarcely addressed in Italy. This study aimed to evaluate the incidence of neoplastic lesions at a scheduled endoscopic follow-up and to identify the patients at higher risk of recurrence. METHODS: Colorectal cancer patients diagnosed in the three participating hospitals (one North, one Centre and one South Italy) were scheduled for colonoscopies at 1, 3 and 5 years after surgery. Incidence of adenomas, advanced adenomas and colorectal cancer was assessed in all patients. Neoplastic incidence in patients with and without synchronous lesions at entry was also compared. RESULTS: Overall, 318 consecutive patients were prospectively enrolled including 108 (34%, group A) with a synchronous lesion and 210 (group B) without it. A cumulative neoplastic incidence of 20.1, 32.4 and 44% was observed at 1, 3 and 5 years of follow-up, respectively. The cumulative incidence of all the lesions was 70% in group A and 30.2% in group B at 5-year follow-up, being 39.5 and 15.5% after excluding the lesions detected at 1-year examination. A neoplastic lesion was detected more frequently in group A at 1year (30.5% versus 14.7%; p = 0.0013), 3 years (21.4% versus 7.6%; p = 0.0008) and at 5years (18.1% versus 7.8%; p = 0.02). CONCLUSIONS: Our data showed that the incidence of adenomas in patients operated for colorectal cancer is fairly high. Colorectal cancer patients with synchronous lesions are at higher risk of neoplastic recurrence at follow-up as compared to those without them. PMID- 16216567 TI - Qualitative research in inflammatory bowel disease: dispelling the myths of an unknown entity. AB - The potential for qualitative research in the field of inflammatory bowel disease has been overlooked due to a misunderstanding of the methods and intent of this approach. In this paper, we provide a brief overview of qualitative research and address some of the misconceptions and criticisms that we have come across during our use of qualitative research methods in inflammatory bowel disease research. PMID- 16216568 TI - Two-step pathway to aminoacylated tRNA. AB - The 3.0 Angstrom crystal structure of a tRNA-dependent amidotransferase from the hyperthermophilic archaeon Pyrococcus abyssi provides the first detailed insight into how cells lacking canonical tRNA synthetases nonetheless carry out protein synthesis with high fidelity. PMID- 16216569 TI - One channel: open and closed. AB - Structural information about the prokaryotic KirBac3.1 inward rectifier family K(+) channel from Magnetospirillum magnetotacticum is reported. These results from two-dimensional electron cryomicroscopy (EM) shed light on the gating mechanism of members of the Kir channel family. PMID- 16216570 TI - Bending in the right direction. AB - The success of a new Monte Carlo algorithm in describing sequence-specific DNA bending is reported. The approach offers the possibility of improved treatments of "indirect readout" effects in the prediction of transcription factor binding sites. PMID- 16216571 TI - YjjX: from structure "Tu" function. AB - It has been shown by structural analysis that YjjX, a hypothetical protein in E. coli, is an ITPase/XTPase and suggest that it may play dual roles in prokaryotic translational regulation and oxidative cell stress response. PMID- 16216572 TI - Structure and function of argonaute proteins. AB - Argonaute (Ago) family proteins are multidomain proteins expressed in prokaryotic and eukaryotic organisms. In eukaryotes, Ago proteins are most well known for their roles in RNA silencing. In prokaryotes, the functions of Ago proteins are unknown, but based on their similarity to eukaryotic Ago proteins, they could be involved in nucleic acid-directed regulatory pathways related to RNA silencing. Recent structural and biochemical studies have shed new light on the function of this family of proteins. These studies reveal how these proteins recognize and cleave RNA and suggest a function for prokaryotic family members. PMID- 16216573 TI - Raman optical activity: a tool for protein structure analysis. AB - On account of its sensitivity to chirality, Raman optical activity (ROA), measured here as the intensity of a small, circularly polarized component in the scattered light using unpolarized incident light, is a powerful probe of protein structure and behavior. Protein ROA spectra provide information on secondary and tertiary structures of polypeptide backbones, backbone hydration, and side chain conformations, and on structural elements present in unfolded states. This article describes the ROA technique and presents ROA spectra, recorded with a commercial instrument of novel design, of a selection of proteins to demonstrate how ROA may be used to readily distinguish between the main classes of protein structure. A principal component analysis illustrates how the many structure sensitive bands in protein ROA spectra are favorable for applying pattern recognition techniques to determine structural relationships between different proteins. PMID- 16216575 TI - Intermediate states of ribonuclease III in complex with double-stranded RNA. AB - Bacterial ribonuclease III (RNase III) can affect RNA structure and gene expression in either of two ways: as a processing enzyme that cleaves double stranded (ds) RNA, or as a binding protein that binds but does not cleave dsRNA. We previously proposed a model of the catalytic complex of RNase III with dsRNA based on three crystal structures, including the endonuclease domain of RNase III with and without bound metal ions and a dsRNA binding protein complexed with dsRNA. We also reported a noncatalytic assembly observed in the crystal structure of an RNase III mutant, which binds but does not cleave dsRNA, complexed with dsRNA. We hypothesize that the RNase III*dsRNA complex can exist in two functional forms, a catalytic complex and a noncatalytic assembly, and that in between the two forms there may be intermediate states. Here, we present four crystal structures of RNase III complexed with dsRNA, representing possible intermediates. PMID- 16216574 TI - Structural basis for tRNA-dependent amidotransferase function. AB - Besides direct charging of tRNAs by aminoacyl-tRNA synthetases, indirect routes also ensure attachment of some amino acids onto tRNA. Such routes may explain how new amino acids entered into protein synthesis. In archaea and in most bacteria, tRNA(Gln) is first misaminoacylated by glutamyl-tRNA synthetase. Glu-tRNA(Gln) is then matured into Gln-tRNA(Gln) by a tRNA-dependent amidotransferase. We report the structure of a tRNA-dependent amidotransferase-that of GatDE from Pyrococcus abyssi. The 3.0 A resolution crystal structure shows a tetramer with two GatD molecules as the core and two GatE molecules at the periphery. The fold of GatE cannot be related to that of any tRNA binding enzyme. The ammonium donor site on GatD and the tRNA site on GatE are markedly distant. Comparison of GatD and L asparaginase structures shows how the motion of a beta hairpin region containing a crucial catalytic threonine may control the overall reaction cycle of GatDE. PMID- 16216576 TI - Crystal structure of human Taspase1, a crucial protease regulating the function of MLL. AB - Taspase1 catalyzes the proteolytic processing of the mixed lineage leukemia (MLL) nuclear protein, which is required for maintaining Hox gene expression patterns. Chromosomal translocations of the MLL gene are associated with leukemia in infants. Taspase1, a threonine aspartase, is a member of the type 2 asparaginase family, but is the only protease in this family. We report here the crystal structures of human activated Taspase1 and its proenzyme, as well as the characterization of the effects of mutations in the active site region using a newly developed fluorogenic assay. The structure of Taspase1 has significant differences from other asparaginases, especially near the active site. Mutation of the catalytic nucleophile, Thr234, abolishes autocatalytic processing in cis but does not completely block proteolysis in trans. The structure unexpectedly showed the binding of a chloride ion in the active site, and our kinetic studies confirm that chlorides ions are inhibitors of this enzyme at physiologically relevant concentrations. PMID- 16216577 TI - Structural basis for glycogen recognition by AMP-activated protein kinase. AB - AMP-activated protein kinase (AMPK) coordinates cellular metabolism in response to energy demand as well as to a variety of stimuli. The AMPK beta subunit acts as a scaffold for the alpha catalytic and gamma regulatory subunits and targets the AMPK heterotrimer to glycogen. We have determined the structure of the AMPK beta glycogen binding domain in complex with beta-cyclodextrin. The structure reveals a carbohydrate binding pocket that consolidates all known aspects of carbohydrate binding observed in starch binding domains into one site, with extensive contact between several residues and five glucose units. beta cyclodextrin is held in a pincer-like grasp with two tryptophan residues cradling two beta-cyclodextrin glucose units and a leucine residue piercing the beta cyclodextrin ring. Mutation of key beta-cyclodextrin binding residues either partially or completely prevents the glycogen binding domain from binding glycogen. Modeling suggests that this binding pocket enables AMPK to interact with glycogen anywhere across the carbohydrate's helical surface. PMID- 16216578 TI - Two different conformational states of the KirBac3.1 potassium channel revealed by electron crystallography. AB - Potassium channels allow the selective flow of K(+) ions across membranes. In response to external gating signals, the potassium channel can move reversibly through a series of structural conformations from a closed to an open state. 2D crystals of the inwardly rectifying K(+) channel KirBac3.1 from Magnetospirillum magnetotacticum have been captured in two distinct conformations, providing "snap shots" of the gating process. Analysis by electron cryomicroscopy of these KirBac3.1 crystals has resulted in reconstructed images in projection at 9 A resolution. Kir channels are tetramers of four subunits arranged as dimers of dimers. Each subunit has two transmembrane helices (inner and outer). In one crystal form, the pore is blocked; in the other crystal form, the pore appears open. Modeling based on the KirBac1.1 (closed) crystal structure shows that opening of the ion conduction pathway could be achieved by bending of the inner helices and significant movements of the outer helices. PMID- 16216579 TI - Metalloproteomics: high-throughput structural and functional annotation of proteins in structural genomics. AB - A high-throughput method for measuring transition metal content based on quantitation of X-ray fluorescence signals was used to analyze 654 proteins selected as targets by the New York Structural GenomiX Research Consortium. Over 10% showed the presence of transition metal atoms in stoichiometric amounts; these totals as well as the abundance distribution are similar to those of the Protein Data Bank. Bioinformatics analysis of the identified metalloproteins in most cases supported the metalloprotein annotation; identification of the conserved metal binding motif was also shown to be useful in verifying structural models of the proteins. Metalloproteomics provides a rapid structural and functional annotation for these sequences and is shown to be approximately 95% accurate in predicting the presence or absence of stoichiometric metal content. The project's goal is to assay at least 1 member from each Pfam family; approximately 500 Pfam families have been characterized with respect to transition metal content so far. PMID- 16216580 TI - Functional plasticity in the substrate binding site of beta-secretase. AB - The aspartic protease beta-secretase (BACE) cleaves the amyloid precursor protein into a 42 residue beta-peptide, which is the principal biochemical marker of Alzheimer's disease. Multiple explicit-water molecular dynamics simulations of the apo and inhibitor bound structures of BACE indicate that both open- and closed-flap conformations are accessible at room temperature and should be taken into account for inhibitor design. Correlated motion is observed within each of the two lobes of BACE, as well as for the interfacial region. A self-inhibited conformation with the side chain of Tyr71 occupying the S(1) pocket is present in some of the unbound simulations. The reversible loss of the side chain hydrogen bond between the catalytic Asp32 and Ser35, due to the concomitant reorientation of the Ser35 hydroxyl group and a water molecule conserved in pepsin-like enzymes, provides further evidence for the suggestion that Ser35 assists in proton acceptance and release by Asp32 during catalysis. PMID- 16216581 TI - Structural and energetic origins of sequence-specific DNA bending: Monte Carlo simulations of papillomavirus E2-DNA binding sites. AB - DNA bending is an important structural feature for indirect readout in protein DNA recognition. The binding of papillomavirus E2 transcription factors to their DNA binding sites is associated with DNA bending, providing an attractive model system to study the origins of sequence-specific DNA bending. The consensus E2 target is of the general form ACCGN(4)CGGT with a variable four base pair region. We applied a new all-atom Monte Carlo (MC) algorithm that combines effective sampling with fast conformational equilibration. The resulting MC ensembles resemble the corresponding high-resolution crystal structures very well. Distinct bending is observed for the E2-DNA binding site with a central AATT linker in contrast to an essentially straight DNA with a central ACGT linker. Contributions of specific base pair steps to the overall bending are shown in terms of local structural parameters. The analysis of conformational substates provides new insights into the energetic origins of intrinsic DNA bending. PMID- 16216582 TI - Identification of an ITPase/XTPase in Escherichia coli by structural and biochemical analysis. AB - Inosine triphosphate (ITP) and xanthosine triphosphate (XTP) are formed upon deamination of ATP and GTP as a result of exposure to chemical mutagens and oxidative damage. Nucleic acid synthesis requires safeguard mechanisms to minimize undesired lethal incorporation of ITP and XTP. Here, we present the crystal structure of YjjX, a protein of hitherto unknown function. The three dimensional fold of YjjX is similar to those of Mj0226 from Methanococcus janschii, which possesses nucleotidase activity, and of Maf from Bacillus subtilis, which can bind nucleotides. Biochemical analyses of YjjX revealed it to exhibit specific phosphatase activity for inosine and xanthosine triphosphates and have a possible interaction with elongation factor Tu. The enzymatic activity of YjjX as an inosine/xanthosine triphosphatase provides evidence for a plausible protection mechanism by clearing the noncanonical nucleotides from the cell during oxidative stress in E. coli. PMID- 16216583 TI - Retroviral matrix domains share electrostatic homology: models for membrane binding function throughout the viral life cycle. AB - The matrix domain (MA) of Gag polyproteins performs multiple functions throughout the retroviral life cycle. MA structures have an electropositive surface patch that is implicated in membrane association. Here, we use computational methods to demonstrate that electrostatic control of membrane binding is a central characteristic of all retroviruses. We are able to explain a wide range of experimental observations and provide a level of quantitative and molecular detail that has been inaccessible to experiment. We further predict that MA may exist in a variety of oligomerization states and propose mechanistic models for the effects of phosphoinositides and phosphorylation. The calculations provide a conceptual model for how non-myristoylated and myristoylated MAs behave similarly in assembly and disassembly. Hence, they provide a unified quantitative picture of the structural and energetic origins of the entire range of MA function and thus enhance, extend, and integrate previous observations on individual stages of the process. PMID- 16216584 TI - Inter- and intramolecular determinants of the specificity of single-stranded DNA binding and cleavage by the F factor relaxase. AB - The TraI protein of conjugative plasmid F factor binds and cleaves a single stranded region of the plasmid prior to transfer to a recipient. TraI36, an N terminal TraI fragment, binds ssDNA with a subnanomolar K(D) and remarkable sequence specificity. The structure of the TraI36 Y16F variant bound to ssDNA reveals specificity determinants, including a ssDNA intramolecular 3 base interaction and two pockets within the protein's binding cleft that accommodate bases in a knob-into-hole fashion. Mutagenesis results underscore the intricate design of the binding site, with the greatest effects resulting from substitutions for residues that both contact ssDNA and stabilize protein structure. The active site architecture suggests that the bound divalent cation, which is essential for catalysis, both positions the DNA by liganding two oxygens of the scissile phosphate and increases the partial positive charge on the phosphorus to enhance nucleophilic attack. PMID- 16216586 TI - Crystal structures of the Mnk2 kinase domain reveal an inhibitory conformation and a zinc binding site. AB - Human mitogen-activated protein kinases (MAPK)-interacting kinases 1 and 2 (Mnk1 and Mnk2) target the translational machinery by phosphorylation of the eukaryotic initiation factor 4E (eIF4E). Here, we present the 2.1 A crystal structure of a nonphosphorylated Mnk2 fragment that encompasses the kinase domain. The results show Mnk-specific features such as a zinc binding motif and an atypical open conformation of the activation segment. In addition, the ATP binding pocket contains an Asp-Phe-Asp (DFD) in place of the canonical magnesium binding Asp-Phe Gly (DFG) motif. The phenylalanine of this motif sticks into the ATP binding pocket and blocks ATP binding as observed with inhibitor bound and, thus, inactive p38 kinase. Replacement of the DFD by the canonical DFG motif affects the conformation of Mnk2, but not ATP binding and kinase activity. The results suggest that the ATP binding pocket and the activation segment of Mnk2 require conformational switches to provide kinase activity. PMID- 16216587 TI - Human DNA polymerase iota incorporates dCTP opposite template G via a G.C + Hoogsteen base pair. AB - Human DNA polymerase iota (hPoliota), a member of the Y family of DNA polymerases, differs in remarkable ways from other DNA polymerases, incorporating correct nucleotides opposite template purines with a much higher efficiency and fidelity than opposite template pyrimidines. We present here the crystal structure of hPoliota bound to template G and incoming dCTP, which reveals a G.C + Hoogsteen base pair in a DNA polymerase active site. We show that the hPoliota active site has evolved to favor Hoogsteen base pairing, wherein the template sugar is fixed in a cavity that reduces the C1'-C1' distance across the nascent base pair from approximately 10.5 A in other DNA polymerases to 8.6 A in hPoliota. The rotation of G from anti to syn is then largely in response to this curtailed C1'-C1' distance. A G.C+ Hoogsteen base pair suggests a specific mechanism for hPoliota's ability to bypass N(2)-adducted guanines that obstruct replication. PMID- 16216585 TI - Features of reovirus outer capsid protein mu1 revealed by electron cryomicroscopy and image reconstruction of the virion at 7.0 Angstrom resolution. AB - Reovirus is a useful model for addressing the molecular basis of membrane penetration by one of the larger nonenveloped animal viruses. We now report the structure of the reovirus virion at approximately 7.0 A resolution as obtained by electron cryomicroscopy and three-dimensional image reconstruction. Several features of the myristoylated outer capsid protein mu1, not seen in a previous X ray crystal structure of the mu1-sigma3 heterohexamer, are evident in the virion. These features appear to be important for stabilizing the outer capsid, regulating the conformational changes in mu1 that accompany perforation of target membranes, and contributing directly to membrane penetration during cell entry. PMID- 16216588 TI - Pulse-pressure-driven neutral lipid accumulation and correlative proinflammatory markers of accelerated atherogenesis. AB - In athero-prone areas of the human aorta, an accumulation of neutral lipids in the arterial intima precedes the arrival of monocytes and the initiation of the fatty streak. This study continues the investigation of a similar preinflammatory process in the coronary arteries of the sodium-sensitive hypertensive Dahl S rat, focusing on the hemodynamic forces associated with the neutral lipid influx and on concurrent changes in serum levels of certain proinflammatory markers. Animals were conditioned from weaning on a high-saturated fat, high-cholesterol diet until early adolescence (age 12 weeks), at which point dietary sodium was increased, and changes in blood pressure, serum analytes and histologic markers were tracked. The first half of the 12-day induction period was characterized by an immediate rise in pulse pressure that persisted above baseline. Serum levels of free monomeric C-reactive protein (mCRP) (as monomer/dimer ratio by Western blot densitometry) rose concurrently with pulse pressure and preceded the influx of neutral lipid into the arterial intima. Levels of oxidized LDL-cholesterol (oxLDL) closely tracked that of neutral lipid accumulation. In this model, elevated pulse pressure appears to drive the activation of circulating CRP and the influx of neutral lipids into the arterial intima, which leads to increased oxLDL levels. PMID- 16216589 TI - Macrophage foam-cell formation in streptozotocin-induced diabetic mice: stimulatory effect of glucose. AB - BACKGROUND: Diabetes is associated with an increased risk for atherosclerosis. We investigated the effect of diabetes induction on atherogenesis and on macrophage foam cell formation. METHODS AND RESULTS: Atherosclerotic apolipoprotein-E deficient mice were converted into diabetic mice by streptozotocin injection. Aortic atherosclerotic lesion area was significantly enhanced by 67% and 106% in mice that were diabetic for 1 and 3 months, respectively, compared to the non diabetic mice. Moreover, mouse peritoneal macrophages (MPM) from diabetic mice for 1 and 3 months exhibit higher lipid peroxides content by 55% and 63%, respectively, in association with the MPM glucose content. Oxidized LDL (Ox-LDL) uptake by MPM obtained from diabetic mice for 1 and 3 months was significantly increased by 36% and 45%, respectively, in association with the increased macrophage cholesterol content. To determine whether the accelerated foam cell formation in diabetic mice could result from a direct effect of glucose on macrophages, J-774-A.1 macrophages were incubated with increasing glucose concentrations (2.5-62 mM). Glucose-enriched macrophages exhibit dose-dependent higher peroxides content up to 7.5-fold and increased Ox-LDL cellular uptake associated with up-regulation of the scavenger receptor CD36 at the mRNA level. CONCLUSION: Induction of diabetes in atherosclerotic mice led to an accelerated atherosclerosis and macrophage-derived foam cell formation, probably by involving a glucose-dependent related mechanism. PMID- 16216590 TI - Glimepiride induces nitric oxide production in human coronary artery endothelial cells via a PI3-kinase-Akt dependent pathway. AB - Diabetes mellitus is one of the major risk factors for coronary artery disease (CAD). A recent study reported that glimepiride, a new third-generation sulfonylurea, inhibited the formation of atheromatous plaques in high-cholesterol fed rabbits. However, the mechanism by which glimepiride induces atheroprotection remains unknown. In the present study, we tested the hypothesis that glimepiride may stimulate NO production in vascular endothelial cells. Human coronary artery endothelial cells (HCAECs) were treated with glimepiride, glibenclamide or vehicle, and NO release was measured. Akt phosphorylation was evaluated by Western blot. The effects of LY294002, a specific PI3-kinase inhibitor, and antisense oligonucleotides directed to Akt, on glimepiride-induced NO production were examined. Glimepiride (0.1-10 microM), but not glibenclamide, induced NO production, significantly increasing it by 1.8-fold (n=6, p<0.05). LY294002 inhibited glimepiride-induced NO production by 68%. Akt was rapidly phosphorylated by glimepiride and antisense oligonucleotides directed to Akt completely inhibited glimepiride-induced NO production. These data demonstrate that glimepiride induces NO production in HCAECs by activating PI3-kinase and Akt, and also suggest that use of glimepiride in type 2 diabetes may show promise for preventing CAD in addition to lowering glucose levels. PMID- 16216592 TI - Smoking, homocysteine and degree of arteriolar retinopathy. AB - It has been demonstrated that higher degree of arteriolar retinopathy is associated with greater cardiovascular risk, and hyperhomocysteinaemia is also related to increased cardiovascular risk, but interacts with other risk factors, particularly smoking. It still remains unclear regarding relationships of smoking, fasting plasma total homocysteine (tHcy) levels and arteriolar retinopathy. This study was aimed to investigate the relationship and influence of smoking and tHcy levels on degree of arteriolar retinopathy. Two hundred and forty-three subjects were enrolled from an annual health examination. The arteriolar retinopathy was examined by direct ophthalmoscopy. Dundett ANOVA showed that geometric mean of tHcy levels were 11.5+/-1.54 versus 11.2+/-1.41 versus 17.6+/-1.92 (P1=0.883, P2=0.001) in subjects with no arteriolar retinopathy (as control group), grades I and II retinopathy, respectively. Furthermore, multiple linear regression analysis showed that only smoking consumption (P<0.001), gender (P=0.012) and presence of hypertension (P=0.041) were independent determinants of plasma tHcy levels. After females were excluded, T-test showed a significant differences in tHcy levels (15.6+/-1.56 micromol/L versus 12.4+/-1.45 micromol/L, P=0.003) and in prevalence of grade II retinopathy (25.4% versus 9.3%, P=0.029), but no difference in other variables or prevalence of overall retinopathy between smokers and non-smokers. Finally, logistic regression showed that smoking (OR 4.19, 95% CI 1.17-15.0) was a stronger predictor than hyperhomocysteinaemia (OR 2.14, 95% CI 0.85-5.41) for presence of grade II retinopathy. This study showed that smoking was related to increased plasma tHcy levels in subjects with grade II retinopathy, and it could independently contribute to facilitating the progression of arteriolar retinopathy. PMID- 16216591 TI - Aspirin inhibits thrombin action on endothelial cells via up-regulation of aminopeptidase N/CD13 expression. AB - OBJECTIVE: We hypothesized that aspirin may exhibit its anti-atherosclerotic effects via mechanisms other than cyclooxygenase inhibition in platelets. METHODS AND RESULTS: Using enhanced subtraction hybridization analysis, we found in human umbilical vein endothelial cells (HUVECs) that aspirin up-regulates the expression of aminopeptidase N (APN/CD13) mRNA and its surface protein levels in a dose-dependent manner. Enzymatic activity of APN/CD13 on HUVECs was increased approximately 1.5-fold by 1 mmol L(-1) of aspirin, and treatment with bestatin, an inhibitor for APN/CD13 metalloprotease activity, attenuated the enhanced activities of APN/CD13. Since activated thrombin receptor is reported to be inactivated by APN/CD13 in vitro, protective actions of aspirin on HUVECs by thrombin stimulation were examined, resulting in the suppression of endothelin-1 and reactive oxygen species productions in HUVECs. These inhibitory actions of aspirin were partially abrogated by bestatin. CONCLUSIONS: Aspirin may exert its anti-atherothrombotic effects in part via the inhibition of thrombin action by up regulating APN/CD13 on endothelial cells. PMID- 16216593 TI - Acute Chlamydia pneumoniae infection with heat-shock-protein-60-related response in patients with acute coronary syndrome. AB - Recent evidence has suggested an association between Chlamydia pneumoniae infection and coronary atherosclerosis. A significant association has also been detected between heat shock protein (HSP) 60 antibody and the severity of coronary atherosclerosis. The aim of this study was to define the relationship between instability of ischemic heart disease (IHD) and serum levels of HSP60 and C. pneumoniae antibodies. Blood samples for the measurement of serum antibody titers were obtained from 1131 patients with ischemic heart disease (65+/-9 years; male/female, 828/303) and 127 non-IHD controls with normal coronary arteries (64+/-9 years; male/female, 60/67) on the day of cardiac catheterization. The serum levels of anti-human HSP60 IgG antibody and anti chlamydial IgM, but not IgG or IgA, antibody were significantly higher in ACS patients than in stable IHD patients or controls. These results suggest that acute C. pneumoniae infection with HSP60-related immunological responses may contribute to the pathophysiology of acute coronary syndromes. PMID- 16216594 TI - Cardiovascular outcome of patients with abnormal coronary vasomotion and normal coronary arteriography is worse in type 2 diabetes mellitus than in arterial hypertension: a 10 year follow-up study. AB - Diabetes and arterial hypertension are major cardiovascular risk factors. Coronary endothelial dysfunction is frequently observed in diabetic and hypertensive patients. This study was designed to compare cardiovascular outcome of hypertensive (HT) and type 2 diabetic patients (D2) with angiographically normal coronary arteries on the basis of their epicardial coronary endothelial function. Coronary reactivity assessment by cold-pressor test (CPT) using quantitative coronary angiography was achieved in 65 HT (45 males, 20 females) aged 51.9+/-7.6 years, and in 59 D2 (32 males, 27 females) aged 48.9+/-7.3 years, with angiographically normal coronary arteries and without other major coronary risk factor. Cardiovascular events (CVE) were recorded with a mean follow-up of 108+/-15 months in HT, and 113+/-10 months in D2. During CPT, in HT coronary artery dilation occurred in 10.8% of the patients, no change in 21.5%, and constriction in 67.7%. In D2, dilation occurred in 3.4% of the patients, no change in 18.6%, and constriction in 78.0%. During follow-up, in HT there were nine CVE in 6/65 patients (9.2%), all in the 6/44 (13.6%) patients with coronary artery constriction. In D2, there were 18 CVE in 16/59 patients (27.1%, P<0.01 versus HT), with 17 CVE in the 15/46 patients with coronary artery constriction, and one CVE in the 1/13 patients without constriction (32.6% versus 7.7%). In patients with coronary artery constriction, CVE were more frequent in D2 than in HT (P<0.05). Last, CVE were more severe and occurred earlier in D2 than in HT. In conclusion, epicardial coronary endothelial dysfunction is predictive of long term CVE in HT and D2 with angiographically normal coronary arteries. Cardiovascular outcome of patients with coronary constriction is worse in D2 than in HT. At the opposite, patients without constriction have good cardiovascular prognosis in both subgroups. PMID- 16216595 TI - Antibodies against oxidized LDL and cardiolipin and mortality in patients with coronary heart disease. AB - The association between antibodies against oxidized LDL (oxLDL) and cardiolipin and the risks of death and cardiovascular disease events were evaluated in patients with established coronary heart disease (CHD). The patients (mean age: 61 years, range: 33-74 years) were participants in the EUROASPIRE study; 108 of them had undergone coronary artery bypass surgery, 106 had balloon angioplasty, 101 had a diagnosis of acute myocardial infarction (AMI), and 98 acute myocardial ischemia. Antibodies against oxLDL and cardiolipin were measured and incidence of CHD events and deaths were followed up for 5 years in 284 men and 129 women. During the follow-up 36 patients died and 21 had AMI. After adjustment for cardiovascular disease risk factors the relative risks (RR [95% confidence interval]) of death were 1 (reference), 2.50 (0.97-6.49) and 2.21 (0.85-5.80) in increasing tertile categories of anti-oxLDL antibody titers, respectively (P for trend 0.16). The risks of CHD-death or AMI combined were 1 (reference), 2.61 (1.02-6.65) and 1.06 (0.37-3.03) in increasing tertile categories of anticardiolipin antibody titers, respectively (P for trend 0.03). In conclusion, the results suggest that antibodies against oxLDL and cardiolipin are not major predictors of risks of death and CHD events in patients with established CHD. PMID- 16216596 TI - A dietary pattern that lowers oxidative stress increases antibodies to oxidized LDL: results from a randomized controlled feeding study. AB - BACKGROUND: Oxidation of LDL (oxLDL) is thought to have an important role in early stages of atherogenesis. Antibody to oxLDL (Ab-oxLDL) has been proposed as a biomarker which might be directly associated with oxidative stress. Yet studies designed to test this hypothesis are lacking. We tested the hypothesis that consumption of a healthy diet rich in fruits and vegetables and reduced in saturated fat, total fat, and cholesterol will concomitantly reduce oxidative stress and Ab-oxLDL. METHODS: One hundred and three healthy individuals were randomly assigned to consume a typical American (control) diet or the DASH diet rich in fruits, vegetables and low-fat dairy products and reduced in fat (27%), saturated fat (7%), and cholesterol (150 mg/day) for 3 months. Outcomes were urinary isoprostanes (in vivo marker of oxidative stress), oxygen radical absorbing capacity (ORAC, an in vitro assay measuring antioxidant activity in serum), and Ab-oxLDL measured at baseline, 1-3 months of feeding. RESULTS: Compared to the control diet, consumption of the DASH diet significantly lowered urinary isoprostane (-226 pg/ml, 95% CI: -420 to -32, P=0.023). Compared with the control group, change in ORAC was higher in the DASH group, 143 trolox units/ml (95% CI: -23 to 308, P=0.091). In comparison with the control diet, increased titers of Ab-oxLDL (37 mU/ml [95% CI: 16-57, P=0.006]) were seen after consumption of the DASH diet. Higher titers of Ab-oxLDL occurred at month 2 (56 mU/ml, 95% CI: 20-90, P=0.004) and month 3 (41 mU/ml, 95% CI: -6 to 88, P=0.082), after initially small increases at month 1 (20 mU/ml, 95% CI: -10 to 51, P=0.176). End-of-study increases in AB-oxLDL were highly correlated with increased ORAC (Spearman's rho=0.46, P<0.0001), but not with changes in specific carotenoids, tocopherols or with change in LDL cholesterol (each: P>0.10). CONCLUSION: Consumption of a healthy diet replete in antioxidants reduced oxidative stress (urinary isoprostanes) yet increased Ab-oxLDL. This indirect association of Ab-oxLDL with urinary isoprostanes hinders use of Ab-oxLDL as a marker of oxidative damage. PMID- 16216598 TI - Rethinking page charges. PMID- 16216601 TI - Prehospital triage of acute myocardial infarction: wireless transmission of electrocardiograms to the on-call cardiologist via a handheld computer. AB - BACKGROUND: Use of intravenous fibrinolytic agents and percutaneous coronary interventions produce the greatest benefit when they are implemented in the first 2 hours after symptom onset. Further delays in the time to treatment typically lead to reduced benefits and poorer outcomes. METHODS: Cabarrus County Emergency Medical Service personnel complete an acute myocardial infarction case report form and assess a 12-lead electrocardiogram (ECG) to determine if ST elevation of at least 1 mV in at least 2 contiguous leads is present and then to transmit the ECG wirelessly to the emergency department (ED). The ECG is then forwarded wirelessly from the ED to the on-call cardiologist who is carrying the IPAQ handheld computer. RESULTS: Five representative patients managed using this system during the initial year of its implementation are presented. CONCLUSION: The examples included in this article illustrate that the system and technology can work if applied in a coordinated fashion using multiple disciplines including emergency medical service, cardiologists, ED personnel, and the hospital cardiac care team, which includes the catheterization laboratory call team, acute coronary care nurses, and clerical support staff. PMID- 16216602 TI - ECGScan: a method for conversion of paper electrocardiographic printouts to digital electrocardiographic files. AB - BACKGROUND: Measurements of parameters from electrocardiograms (ECGs) are still largely performed from paper ECG records. Recent guidelines from regulatory agencies and, in particular, the requirement of the Food and Drug Administration to enforce the digital submission of annotated ECGs have triggered significant efforts in the pharmaceutical industry, which, to comply with the new guidelines, is adopting digital ECG technology. At the same time, the new requirements justify the need for tools to convert existing paper ECG records into digital format, particularly for retrospective studies. METHODS: This article presents ECGScan, a computer application developed for the conversion of paper ECG records to digital ECG files. An image processing engine is used to first detect the underlying grid and, subsequently, to extrapolate the ECG waveforms using a technique based on active contour modeling. RESULTS: ECGScan was validated using a set of 60 ECGs for which both the original digital waveform and paper printouts were available. Sample-by-sample comparisons provided evidence of a robust wave reconstruction (root mean square value from 169 PQRST complexes was 16.8+/-11.8 microV). Semiautomatic measurements of QT intervals performed on 144 complexes also indicated a strong agreement between original and derived ECGs (DeltaQT=0.577+/-5.41 milliseconds). CONCLUSIONS: ECGScan provides a robust reconstruction of a digital ECG, both in waveform reconstruction and in QT measurements performed on original (digital) ECGs and on digitized ECGs from paper printouts. PMID- 16216603 TI - Points to consider in electrocardiogram waveform extraction. PMID- 16216604 TI - Electrocardiogram digitization: a practical perspective on the usefulness of a new tool to convert paper electrocardiograms into digital waveform. PMID- 16216605 TI - Widely split P waves in the presence of interatrial block. AB - Interatrial block (IAB; P wave>or=110 milliseconds), conduction delay between the atria, is highly prevalent and is associated with atrial tachyarrhythmias, left atrial electromechanical dysfunction, as well as a potential risk for systemic embolism. However, much is still yet to be known of IAB's exact pathophysiology and how it may manifest in relation to medical disease. We present an unusual case of widely split P waves in the presence of IAB in a severely ill patient. PMID- 16216606 TI - The prolonged P wave and interatrial block. Time to consider a broader concept and different terminology. PMID- 16216607 TI - Measurement and reproducibility of magnetocardiographic filtered atrial signal in patients with paroxysmal lone atrial fibrillation and in healthy subjects. AB - Magnetocardiography (MCG) is a method complementary to electrocardiography (ECG). We examined recording and reproducibility of atrial depolarization signal by MCG. Multichannel MCG over anterior chest and orthogonal 3-lead ECG were recorded in 9 patients who had paroxysmal lone atrial fibrillation and in 10 healthy subjects in duplicate at least 1 week apart. Data were averaged using atrial wave template and high-pass filtered at 25, 40, and 60 Hz. Atrial signal duration with automatic detection of onset and offset and root mean square amplitudes of the last portion of atrial signal were determined. Coefficient of variation of atrial signal duration by MCG at 40 Hz was 3.3% and difference between the measurements was 3.5 milliseconds on average. The corresponding figures obtained by signal averaged ECG (SAECG) were 6.1% and 6.9 milliseconds. Coefficient of variation for root mean square of the last 40 milliseconds of atrial signal were 16% in MCG and 17% in SAECG. Reproducibility was best at 40-Hz filter and similar in patients and healthy subjects. In conclusion, the reproducibility of atrial signal variables in MCG is adequate and somewhat better than in SAECG and equal in patients with lone atrial fibrillation and healthy subjects. Magnetocardiography seems to be a potentially valuable method to evaluate features of atrial depolarization in patient studies. PMID- 16216608 TI - An unusual electrocardiogram artifact: what is its source? AB - A diabetic female presented with nausea and vomiting. Her electrocardiogram showed sinus rhythm with two artifactual spikes, not synchronized with the cardiac rhythm. The patient had an implanted gastric electrical stimulation system for treating her diabetic gastroparesis. Recent DC shock for ventricular fibrillation during coronary angiography caused malfunction of the gastric pacemaker. PMID- 16216609 TI - Neurocardiogenic syncope in chronic atrioventricular block. AB - We report a 37-year-old man with type I second-degree atrioventricular (AV) block (atypical Wenckebach's periodicity) referred to our department for pacemaker implantation because of an episode of syncope. After exhaustive evaluation, including electrophysiological test, in which Wenckebach's cycles with block within the AV node was demonstrated, syncope was considered to be neurally mediated. Head-up tilt testing with sublingual isosorbide dinitrate was positive. The decrease in atrial rate at the beginning of the vasovagal reaction was not immediately accompanied by a depressed AV node conduction. Only at the moment of syncope did incomplete AV block appear. This observation illustrates (1) a neurally mediated origin of syncope in a patient with chronic AV block, and (2) the different time-course responses of the sinus and AV nodes to autonomic tone. PMID- 16216610 TI - A new look at an old question-how good is the computer in the diagnosis of arrhythmias? PMID- 16216611 TI - Effects of thrombolysis on vectorcardiographic indices of ventricular repolarization: correlation with ST-segment resolution. AB - To investigate the effects of thrombolysis on vectorcardiographic (VCG) descriptors of ventricular repolarization in association with ST-segment resolution, 70 consecutively recruited patients with acute myocardial infarction underwent digital 12-lead electrocardiograms (ECGs) before and at 3 hours after thrombolysis. The alterations in the VCG descriptors spatial T amplitude and spatial QRS-T angle from the pre- to the post-thrombolysis ECG, as well as the ST segment resolution, were calculated. Angiography revealed patency of the infarct related coronary artery after thrombolysis in 52 (74%) patients (group A) and occlusion in 18 (26%) (group B). The spatial T amplitude was highly significantly reduced after thrombolysis in group A (P<.0001), but only marginally reduced in group B (P=.016). The spatial QRS-T angle was also significantly, although only marginally, reduced after thrombolysis in group A (P=.019), whereas it was not changed after thrombolysis in group B (P=.868). An ST-segment resolution of 60% and a 25% reduction in the spatial T amplitude after thrombolysis were able to identify patency of the infarct-related coronary artery with sensitivities of 90% and 77% and specificities of 94% and 74%, respectively. Both VCG descriptors were significantly affected by thrombolysis in patients with acute myocardial infarction, but constituted only moderate markers of thrombolysis efficacy, as evidenced by the presence of patency in the infarct-related coronary artery, compared with the ST-segment resolution. PMID- 16216612 TI - Is the outcomes of early ST-segment resolution after thrombolytic therapy in acute myocardial infarction always favorable? AB - AIMS: To determine whether the magnitude of ST-segment resolution after thrombolytic therapy (TT) predicts short- and long-term outcomes in an unselected population of patients with an acute myocardial infarction (AMI). BACKGROUND: Recent studies suggest that resolution of ST-segment elevation (STE) on the 2 hour post-TT electrocardiogram (ECG) is a useful predictor of prognosis. However, these studies were restricted to clinical trials where only 15% to 20% of the patients receiving TT were often enrolled. METHODS: The present study evaluated an unselected consecutive group of patients who received TT. All clinical, investigational, and follow-up data had been collected in a prospective manner. The analysis of ECGs was done retrospectively with the reader blinded to the clinical course. STE at 80 milliseconds after the J point was measured on the baseline and 90-minute ECG using a hand-held caliper. The resolution of STE was categorized as complete (>or=70%), partial (30% to <70%), and none (<30%) as has been done in previous studies. RESULTS: Three hundred fifty-two patients (250 men, 102 women; age, mean+/-SEM, 61.8+/-1.0 years; peak creatine kinase, 1938+/ 185 micromol/L; door to needle time, 50.0+/-6 minutes, <30 minutes, 50%; <45 minutes, 70%) with AMIs who received TT were included in the study. Inhospital deaths and recurrent AMI/postinfarct angina revealed no significant association with increasing ST-segment resolution (P>.05). A 70% or higher ST-segment resolution was associated with a significantly lower incidence of inhospital congestive heart failure (CHF) and CHF/death (P<.05). Similarly, with a 70% or higher ST-segment resolution, there was a lower incidence in the 1-year outcomes of CHF and death/CHF. However the 1-year occurrences of unstable angina or recurrent AMIs taken singly did not bear a correlation to increasing magnitudes of ST-segment resolution (P>.05). Although as a composite measure, there was an increasing trend with ST-segment resolution. CONCLUSIONS: Magnitude of ST-segment resolution after TT appears to demonstrate a dichotomous relationship to measured outcomes. Although there is a lower incidence of death/CHF with increasing ST segment resolution, there appears to be a higher likelihood for recurrent AMI/unstable angina. PMID- 16216613 TI - Inverted T waves on electrocardiogram: myocardial ischemia versus pulmonary embolism. AB - Electrocardiogram (ECG) is of limited diagnostic value in patients suspected with pulmonary embolism (PE). However, recent studies suggest that inverted T waves in the precordial leads are the most frequent ECG sign of massive PE (Chest 1997;11:537). Besides, this ECG sign was also associated with the best sensitivity, specificity, and positive and negative predictive values for diagnosing PE. We report 2 cases with similar ECG findings that were referred to us as unstable angina. Both were hemodynamically stable and had moderate-size pulmonary emboli. The ECG findings reverted to normal within a week of anticoagulation treatment. Our observation suggests that even a moderate-size PE can cause these ECG changes. PMID- 16216614 TI - On the mechanisms for the conversion of ventricular fibrillation to tachycardia by perfusion with ruthenium red. AB - We have recently demonstrated that during pacing-induced sustained ventricular fibrillation (VF), perfusion of the heart with either ruthenium red (RR) or Ru 360, blockers of the mitochondrial Ca2+ uniporter, resulted in the reversible conversion of VF to ventricular tachycardia (VT). Here, we aimed at elucidating the electrophysiological mechanisms for the RR-induced conversion of VF to VT. The experiments were performed using Langendorff-perfused isolated rat hearts in which left ventricular pressure and left ventricular intracellular action potential were recorded. Perfusion with either RR or Ru 360 resulted in decreases in the action potential duration (APD), refractory period, and slope of APD restitution curves. These changes were antagonized by cotreatment with S(-)-Bay K8644. In addition, perfusion with verapamil produced the decreases in APD at 90% repolarization, refractory period and slope of APD restitution curves similar to the RR or Ru 360 perfusion. Such electrophysiological changes may be responsible for the reversible conversion of sustained VF to VT caused by perfusion with RR or Ru 360. PMID- 16216616 TI - Short QT syndrome--5 years of progress. PMID- 16216615 TI - Short QT interval syndrome: a case report. AB - Twelve-lead electrocardiograms revealed fine atrial fibrillation and a short QT interval (SQTI) (<300 milliseconds) with an average ventricular rate of 54/min in a 20-year-old male presented with exertional dyspnea. His echocardiographic evaluation revealed interatrial septal aneurysm and slightly dilated pulmonary artery. An electrophysiologic study revealed atrial fibrillation with a very high frequency, short ventricular effective refractory period (130 milliseconds) and ventricular fibrillation inducible with 3 short coupled extrastimuli. Signs were consistent with the rare SQTI syndrome. Although SQTI syndrome is associated with increased risk for sudden cardiac death, the patient was free of arrhythmia symptoms and denied any syncope or presyncope. Family history was also negative for sudden cardiac death and for any symptom suggestive of arrhythmia. The patient refused implantable defibrillator and was treated with anticoagulation and quinidine therapy. PMID- 16216617 TI - Prominent positive U waves appearing with high-dose intravenous phenylephrine. AB - A 71-year-old woman who had critical cerebral ischemia secondary to a carotid artery occlusion was receiving high-dose intravenous phenylephrine for a trial of hypertensive therapy. While on a maximal dose of phenylephrine she developed prominent positive U waves, which disappeared with the cessation of the drug. The possible underlying electrophysiological mechanisms of this phenomenon are discussed in this paper. PMID- 16216618 TI - Report of the first virtual visualization of the reconstructed electrocardiographic display symposium. AB - In August 2004, a group of scientists and clinicians with a deep interest in electrocardiography met to discuss the present and future of the electrocardiogram as an imaging modality. Motivated by a set of challenges to the field, they each presented and discussed their ideas about the basic electrophysiology, the computational approaches required, and the clinical state of the art and where it might go in the future. In this paper, we present a summary of these presentations and discussions, starting with a statement of the challenges and a motivating case study that illustrates the inadequacies of electrocardiography as it is current practiced. Following this introduction are overviews of the present state of the inverse problem of electrocardiography and the underlying assumptions of this form of simulation and modeling. We conclude with a summary of the needs that we feel must be addressed to achieve the full potential of electrically based imaging of the heart. PMID- 16216619 TI - Prolonged recovery time in the left precordial leads reflects increased left ventricular mass in the hypertensive patients. AB - Hypertensive left ventricular hypertrophy (LVH) is considered to be a risk for arrhythmogenicity, but the quantification of the changes in T-wave morphology, as the reflection of repolarization abnormality, has not been fully established. The purpose of this study was to quantify the T-wave changes in the hypertensive patients and to investigate the relationship with the increased left ventricular mass. Standard 12-lead electrocardiogram and echocardiogram were recorded in 90 hypertensive patients. Activation time (AT), activation recovery interval (ARI), and recovery time (RT) were measured in the precordial lead and QT interval in the 12 leads. To compare the left precordial T-wave changes among patients, measurements of ARI and RT in the right precordial negative T wave were excluded. Each parameter excluding AT was corrected with Bazett formula, and then the dispersion was calculated. Left ventricular mass index was determined echocardiographically to select non-LVH group (n=31) and LVH group (n=59). In both groups, AT, ARI, and RT in the left precordial leads were larger compared with those in the right precordial leads. Dispersion of AT was not different between the 2 groups. However, the dispersion of ARI and RT in LVH group was significantly greater than that in non-LVH group. There were correlations between left ventricular mass index and the dispersion of RT (r=0.66, P<.001), ARI (r=0.61, P<.001), and 12-lead QT (r=0.42, P<.001). In patients with LVH, significant prolongation of RT in the left precordial leads was observed, suggesting that this RT change resulted from the nonuniformity of epicardial action potential duration. PMID- 16216620 TI - Acute myocardial infarction in a patient with dextrocardia and situs inversus. AB - We report a case of 54-year-old man who presented to hospital with severe prolonged retrosternal chest pain of anginal nature. Electrocardiogram taken by his general practitioner showed minimal ST elevation in chest leads V1 and V2; there was also marked right axis deviation of P wave (negative in lead I and aVL) and of QRS complexes, together with low voltage in precordial leads V4 through V6 suggestive of dextrocardia. Repeat electrocardiogram with chest and limb leads reversed showed widespread, significant ST elevation in lead I, aVL, and V1 through V5 in keeping with extensive acute anterior myocardial infarction (MI). High cardiac enzymes and troponin level provided further confirmation. The extent of MI in such patients may be underestimated unless dextrocardia is timely recognized and leads reversed. We recommend that for patients with dextrocardia and situs inversus presenting with MI, both chest and limb leads be reversed to reveal the true extent of the infracted area. PMID- 16216621 TI - RE: Differentiating an acute MI from ventricular aneurysm in the presence of RBBB (revised). PMID- 16216622 TI - Apparent conversion of complete to incomplete bundle branch block due to peripheral edema. PMID- 16216623 TI - Current issues in Wilms tumor management. PMID- 16216624 TI - Phytoremediation management of selenium-laden drainage sediments in the San Luis Drain: a greenhouse feasibility study. AB - An estimated 100,000m(3) selenium (Se)-laden drainage sediment resides in the San Luis Drain (SLD) of Central California. This greenhouse study was undertaken to evaluate the feasibility of growing salt- and boron-tolerant plant species in sediment for reduction of Se content by plant extraction. Drainage sediment was collected from the SLD and mixed with control soil (i.e., uncontaminated soil) to the following ratios (sediment:control soil) by volume: 0:3 (i.e., control soil only), 1:2 (i.e., 1/3 sediment and 2/3 control soil), 2:1 (i.e., 2/3 sediment and 1/3 control soil), and 3:0 (i.e., sediment only). Salt-tolerant plant species consisted of canola (Brassica napus var. Hyola 420), tall fescue (Festuca arundinacea var. Au Triumph), salado grass (Sporobulus airoides), and cordgrass (Spartina patens var. Flageo). Increased ratios of sediment:soil resulted in decreased dry matter production for all tested plant species; especially at ratios of sediment:soil greater than 1:2. Plant Se concentrations (mgkg(-1) DM) ranged as follows for plant species at all ratios of sediment:soil: canola (51 72), tall fescue (16-36), and cordgrass and salado grass (9-14). Total Se concentrations in the soil were at least 20% lower at postharvest compared to preplant concentrations for all plant species at each ratio of sediment:soil. In contrast, water-extractable Se concentrations in the soil were at least three times higher at postharvest than at preplant for all plant species, irrespective of the ratio of sediment:soil. Leaching of Se occurred in irrigated bare pots from each respective ratio of sediment:soil over a duration of 60 days. Based upon the downward movement of Se in bare pots of sediment:soil, it may be more prudent to leave the drainage sediment in the SLD, incorporate clean soil, and then grow low maintenance salt-tolerant plants (e.g., cordgrass, salado grass) in the concrete-lined canal. By this means, possible contamination of groundwater with soluble Se will be eliminated, while phytoremediation slowly reduces Se content in the drainage sediment. PMID- 16216625 TI - Physiological effects of thermomechanical newsprint mill effluent on Atlantic salmon (Salmo salar L.). AB - Anadramous Atlantic salmon (Salmo salar L.) returning to Exploits River, Newfoundland, Canada, to spawn encounter low concentrations of thermomechanical pulp (TMP) effluent as they migrate upstream past an integrated newsprint mill. Various physiological responses of adult Atlantic salmon from the Exploits River were studied under laboratory conditions. The effects of a 6-h exposure to 0%, 6%, 12%, or 25% (v/v) TMP effluent or an increasing concentration gradient of effluent (0-25%) on cardiac output (Q ), critical swimming performance (U(crit)), hematocrit, and blood glucose, cortisol, lactate, and osmolality were examined. Relative to other treatment groups, Q during routine and low-level activity was 7 10% higher in fish exposed to at least 12% effluent. The 25% exposure group had a distinctly lower U(crit) and scope for increase in Q than the other treatment groups. These findings suggest that effluent exposure elevates physiological maintenance and repair costs, resulting in a reallocation of energetic resources. PMID- 16216626 TI - Development of a short-term in situ caging methodology to assess long-term effects of industrial and municipal discharges on salmon smolts. AB - Trial experiments to develop an in situ method for determining effects of short term exposure to contaminants on salmon during the sensitive smolt stage were carried out. Wild Atlantic salmon (Salmo salar) smolts were caged at various estuarine locations in northeast New Brunswick, Canada at different levels of anthropogenic impact in 2000 and 2001. Survival, growth (weight and K), and feeding (delta(13)C and delta(15)N) parameters were measured following summer grow-out at a hatchery. A summary of distributions of the four parameters indicated that smolts caged at locations receiving lower levels of industrial and municipal discharges feed and grow better than those caged at sites receiving higher levels. The findings, however, were not repeatable between the 2 years studied. The observed inconsistency between years may be a consequence of the relatively low concentrations of alkylphenolic contaminants (putative causative agents) and overall steroidogenic potency in river water at the time of caging. Differences in temperature and salinity from 2000 to 2001 may have further confounded comparisons across treatments and between years. In future studies, caging in closer proximity to industrial and municipal discharges or in systems with higher concentrations of waterborne contaminants or impoundments would help further the assessment and applicability of this methodology and allow a more robust comparison of smolt feeding and growth among reference and exposure sites. PMID- 16216627 TI - Characterization of the cholinesterases present in head tissues of the estuarine fish Pomatoschistus microps: application to biomonitoring. AB - In recent years biomarkers have been widely used for the assessment of effects and/or exposure to environmental contaminants. One of the most frequently used biomarkers is the inhibition of cholinesterases (ChE), which is a useful indicator of organophosphate and carbamate exposure and/or effects. Recent studies indicated that more than one ChE may be present in tissues of fish and that different forms may vary in their sensitivity to anticholinesterase agents. Cholinesterase activity of the juvenile of the common goby (Pomatoschistus microps), a widespread fish in estuaries of the Atlantic coast of northwestern Europe, was characterized using four substrates (acetylthiocholine iodide, acetyl beta-metylthiocholine iodide, propionylthiocholine iodide, and S butyrylthiocholine iodide) and three ChE inhibitors (eserine sulfate, BW284C51, and iso-OMPA) in different tissues of the fish head. In addition, the range of ChE activity that may be considered as "normal" for non-exposed P. microps was determined. The results suggest the presence of two types of ChE in the whole head homogenate. The present study underscores the relevance of ChE characterization before its use as a biomarker in biomonitoring studies. PMID- 16216628 TI - Detection of DNA damage by alkaline single cell gel electrophoresis in 2,4 dichlorophenoxyacetic-acid- and butachlor-exposed erythrocytes of Clarias batrachus. AB - The alkaline single cell gel electrophoresis, also known as comet assay, is a rapid, simple and sensitive technique for measuring DNA strand breaks in individual cells. The present study was undertaken to evaluate the genotoxic potential of two widely used herbicides; 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-diethyl-N-(butoxymethyl) acetanilide (butachlor) in erythrocytes of freshwater catfish, Clarias batrachus. Fish were exposed by medium treatment with three sub-lethal concentrations of 2,4-D (25, 50, and 75ppm) and butachlor (1, 2, and 2.5ppm) and alkaline comet assay was performed on nucleated erythrocytes after 48, 72, and 96h. The amount of DNA damage in cells was estimated from comet tail length as the extent of migration of the genetic material. A significant increase in comet tail length indicating DNA damage was observed at all concentrations of both the herbicides compared with control (P<0.05). The mean comet tail length showed a concentration-related and time dependent increase as the maximum tail length recorded at highest concentration and longer duration of 2,4-D (9.59microm) and butachlor (9.28microm). This study confirmed that the comet assay applied on the fish erythrocyte is a useful tool in determining potential genotoxicity of water pollutants and might be appropriate as a part of a monitoring program. PMID- 16216629 TI - Interleukin-1alpha and tumor necrosis factor alpha modulate cytochrome P450 activities in carp (Cyprinus carpio). AB - In mammals, it has been shown that the activation of host defense mechanisms down regulates microsomal cytochrome P450 by the liberation of cytokines. We investigated the effect of interleukin-1alpha (IL1alpha) and tumor necrosis factoralpha (TNFalpha) on constitutive and 3-methylcholanthrene (3-MC)-induced biotransformation activities in carp. We have first measured the time course response of ethoxyresorufine O-decthylase (EROD) activity in liver, head kidney, and spleen 1, 2, 3, 5, and 7 days after intraperitoneal injection of a prototypical Cyp 1A inducer (3-MC). This activity was compared to the rate of 3 MC accumulation in all organs tested. A correlation between a diminution of EROD activity and an increase in 3-MC concentration in each organ was observed. We have also tested the effects of two inflammatory cytokines (IL1alpha and TNFalpha) on biotransformation activities. Intravenous injection of these compounds resulted in a marked depression of 3-MC-induced glutathione S transferase activity in all organs tested and in 3-MC-increased cytochrome P450 content in the liver and head kidney. TNFalpha produced an increase in basal EROD activity in the liver and head kidney. Taken together, these results suggested that, as in mammals, the activation of host defense mechanisms regulates microsomal cytochrome P450 and related enzymes in fish. PMID- 16216630 TI - Cytochrome P4501A induction and DNA adduct formation in glaucous gulls (Larus hyperboreus), fed with environmentally contaminated gull eggs. AB - This study indicates that complex mixtures of pollutants found in the Arctic marine environment have genotoxic effects in glaucous gulls (Larus hyperboreus). DNA adducts were quantified, by the (32)P-postlabeling technique, in liver samples from gulls fed with hen eggs (controls) and from gulls fed with environmentally contaminated gull eggs (exposed). All birds were grown and fed under laboratory conditions. Hepatic homologues to mammalian cytochrome P4501A (CYP1A) proteins were also determined by Western blotting. DNA adducts were detected in all but one liver sample, but the exposed birds had a significantly increased level of DNA adducts relative to that of the controls. There was no clear significant correlation between the DNA adduct level and the level of organochlorine compounds (OCs) in blood. The level of CYP1A protein was significantly higher in the liver of exposed male gulls than in the liver of control males and positively correlated, with significance, to the level of OC compounds measured in blood. There was no significant correlation between the level of DNA adducts and the CYP1A protein content. PMID- 16216631 TI - Effects of salinity on biomarker responses in Crassostrea rhizophorae (Mollusca, Bivalvia) exposed to diesel oil. AB - Crassostrea rhizophorae is a euryhaline oyster that inhabits mangrove areas, which are widely distributed along the Brazilian coast. The aim of this study was to investigate the effects of salinity (9, 15, 25, and 35ppt) on the activities of glutathione S-transferase (GST), glucose 6-phosphate dehydrogenase (G6PDH), catalase (CAT), and acetylcholinesterase (AChE) in the digestive gland of this species after exposure to diesel oil for 7 days at nominal concentrations of 0.01, 0.1, and 1mlL(-1) and after depuration for 24h and 7 days. GST activity increased in a diesel oil concentration-dependent manner at salinities 25 and 15ppt and remained slightly elevated even after depuration periods of 24h and 7 days. No changes were observed in the activities of G6PDH, CAT, and AChE in the oysters exposed to diesel and depurated. Based on these results, GST activity in the digestive gland of C. rhizophorae might be used as a biomarker of exposure to diesel oil in sites where the salinity is between 15 and 25ppt, values usually observed in mangrove ecosystems. PMID- 16216632 TI - Comparison of porewater and elutriate bivalve larval development toxicity testing in a sediment quality triad framework. AB - Concurrent porewater (PW) and elutriate (ELU) toxicity testing using newly fertilized larvae of the bivalve Mytilus galloprovincialis was conducted as part of sediment quality triad (SQT) investigations for urban harbor locations. PW samples were consistently more toxic to bivalve larvae than the corresponding ELU sample, including samples collected from uncontaminated reference locations. Ammonia was identified as the most likely toxic agent. EC(20) and EC(50) values of 0.028 and 0.036mg/L un-ionized N, respectively, were determined for M. galloprovincialis. The limitations of incorporating PW bivalve larval development toxicity tests using M. galloprovincialis for routine SQT investigations, as well as possible alternative methods, are discussed. PMID- 16216633 TI - Effect of solar UV radiation on earthworm (Metaphire posthuma). AB - Human health risks like damage to the eyes, immune system, and skin are known to be associated with increasing ultraviolet radiation (UVR) in the environment. In this study, we evaluated the phototoxic effects of UVR in sunlight and its possible mechanism of action by using earthworm as an alternative model because earthworm skin contains several biomolecules (tetraene and triene sterol) similar to human beings. We studied the generation of reactive oxygen species (ROS), photooxidation of lipids, and histopathological changes in earthworm integument. It was observed that UVR-exposed earthworm skin homogenate produced a significant amount of singlet oxygen ((1)O(2)), superoxide anion (O(2)(*)(-)), hydroxyl radicals ((*)OH), and photooxidation of lipids. The production of ROS and lipid peroxidation product was found dependent on the dose of solar UVR in earthworm integument. Histological anomalies such as thickening, vacuolation, and hypertrophy of epidermal cells were observed when the animals were exposed for 1 to 2h, while a higher exposure period (3h) caused degeneration of circular and longitudinal muscles. Continuous sunlight exposure for more than 3h was found lethal to worms. These observations suggested that the current level of UVR in sunlight may produce significant phototoxic effects in the earthworms probably via the generation of ROS (photodynamic action). Possible increases in UVR in view of ozone depletion may be more detrimental to the biomolecules in the worm's skin. The earthworm thus turned out as a simple, sensitive, and cost-effective test organism for the assessment of the hazardous potential of solar radiation and also for planning safety measures for human beings. PMID- 16216634 TI - Assessment of the environmental hazard from municipal and industrial wastewater treatment sludge by employing chemical and biological methods. AB - Chemical analyses and toxicity testing were employed in conjunction to evaluate the environmental hazard from the wasted sludge generated during the biological treatment of urban and industrial wastewaters. Chemical analyses included determination of seven polychlorinated biphenyls (PCBs), 13 polycyclic aromatic hydrocarbons (PAHs), total organic carbon (TOC), and seven heavy metals (As, Cd, Cr, Cu, Pb, Mn, and Zn) in sludge and sludge leachates deriving from two standard leaching procedures: (a) the mild leaching test EN-12457-2 proposed by EC and (b) the relatively aggressive toxicity characteristic leaching procedure (TCLP) leaching test proposed by US EPA. Acute toxicity measurements were performed in aqueous sludge elutriates and leachates by using bioluminescence bacteria. The urban sludge was found to be more enriched with PAHs than the industrial sludge, however, at levels below the EU limits for sludge application. The total PCB content (Sigma7PCBs) in both sludges, particularly in the industrial sludge, exceeded the proposed European limit for sludge use as soil amendment. With regards to their heavy metal content, both sludges met the requirements for use in agriculture. The urban sludge exhibited high ecotoxicity, while the industrial sludge with almost two times lower toxicity was classified as not toxic to slightly toxic. The EN and the TCLP leaching procedures resulted in different sludge characterizations both from chemical and from ecotoxicological points of view. The EN procedure appeared to be more sensitive to the potential environmental risk from sludge disposal. The results of the study revealed the necessity for combining chemical with ecotoxicological criteria for integrated characterization of wasted sludge and the need for harmonization of the methods employed for waste classification. PMID- 16216635 TI - Morphological and metabolic alterations in duckweed (Spirodela polyrhiza) on long term low-level chronic UV-B exposure. AB - Laboratory grown duckweed (Spirodela polyrhiza) plants were exposed to 0.72 and 1.44J of UV-B radiation daily for 7 days at 0.4mW/cm(2) intensity. Chlorosis and necrosis were observed along with depletion in protein, pigments (chlorophyll, pheophytin, carotenoids, phycoerythrin, phycocyanin, and flavoxanthin), biomass, root length, and frond size in UV-B-exposed plants. The study confirms morphological and metabolic alterations leading to reduction in the productivity of duckweed following long-term exposure to UV-B radiation. PMID- 16216636 TI - Assessment of genotoxic activity of petroleum hydrocarbon-bioremediated soil. AB - The relationship between toxicity and soil contamination must be understood to develop reliable indicators of environmental restoration for bioremediation. Two bacterial rapid bioassays, SOS chromotest and the umu test with and without metabolic activation (S-9 mixture), were used to evaluate the genotoxicity of petroleum hydrocarbon-contaminated soil following bioremediation treatment. The soil was taken from an engineered biopile at the Czechowice-Dziedzice Polish oil refinery (CZOR). The bioremediation process in the biopile lasted 4 years, and the toxicity measurements were done after this treatment. Carcinogens detected in the soil, polyaromatic hydrocarbons (PAHs), were reduced to low concentrations (2mg/kg dry wt) by the bioremediation process. Genotoxicity was not observed for soils tested with and without metabolic activation by a liver homogenate (S-9 mixture). However, the umu test was more sensitive than the SOS chromotest in the analysis of petroleum hydrocarbon-bioremediated soil. Analytical results of soil used in the bioassays confirmed that the bioremediation process reduced 81% of the petroleum hydrocarbons including PAHs. We conclude that the combined test systems employed in this study are useful tools for the genotoxic examination of remediated petroleum hydrocarbon-contaminated soil. PMID- 16216637 TI - Effect of sulfur dioxide hydrates on cell cycle, sister chromatid exchange, and micronuclei in barley. AB - The effects of sulfur dioxide (SO(2)) hydrates exposure on cell cycle, sister chromatid exchange (SCE), and micronuclei (MN) were investigated in barley (Hordeum vulgare) roots. A mixture of sodium bisulfite and sodium sulfite (1:3), at various concentrations from 1x10(-5) to 3x10(-2)M, was used for the treatments. The results showed that the mixture induced the formation of SCE and MN in barley root cells with different effective concentrations and with different trends as treatment concentrations increased. At high concentrations of 0.5-30.0mM, SO(2) hydrates inhibited the mitotic activity and the growth of barley roots by cell cycle delay and cell death, but at 0.1mM, the chemicals slightly stimulated mitotic activity and root growth. These remarkable effects in causing DNA damage and consequent chromosome damage suggest that SO(2) is genotoxic agent and its genotoxicity may influence the mitotic activity and plant growth under SO(2) stress. PMID- 16216638 TI - Estrogenic effects in crucian carp (Carassius carassius) exposed to treated sewage effluent. AB - To assess the estrogenicity of a municipal sewage treatment plant, sexually mature crucian carp of both sexes were exposed for 28 days to domestic treated sewage effluent running with a constant flow of water, with tap water and with different concentrations of the sewage effluent (25%, 50% and 100%). Vitellogenin (VTG), gonadosomatic index (GSI), hepatosomatic index (HSI), and histological abnormalities were used to assess the estrogenic potency of the effluent. Results show a significant (P<0.05) increase in VTG levels in all exposed male fish. A significant (P<0.05) reduction in the GSI was only observed in fish exposed to 100% effluent. Morphological changes were detected by histological evaluation, revealing severe effects on the testes. Spermatogenesis was progressively reduced to total inhibition in fish exposed to 100% effluent. One of the most pronounced effects detected was the presence of oocytes in male gonads, observed in 20% of males exposed to 100% sewage effluent. PMID- 16216639 TI - Anti-apoptotic action of zearalenone in MCF-7 cells. AB - Zearalenone (ZEA), a nonsteroidal estrogenic mycotoxin, is present in high concentrations in dairy products and cereals. Studies have indicated that ZEA could strongly provoke proliferation in estrogen-dependent breast cancer MCF-7 cells following estrogen ablation. The current study confirmed the previous studies that within the range of concentrations of 2-96nM, like endogenous estradiol, ZEA could stimulate proliferation in MCF-7 cells with inducing a profound increase in S phase and a modest increase in G(2)/M phase that was accompanied by a decrease in G(0)/G(1) phase. The Cell Death Detection ELISA was used to determine whether the robust cell viability retrieved by ZEA was a result of inhibited apoptosis. Data indicated that ZEA-mediated inhibition of apoptosis is significantly evident (P<0.05) and in a dose-dependent manner. Western blot and multiple RT-PCR analysis revealed that the anti-apoptotic bcl-2 was upregulated at both protein and mRNA levels, together with the downregulation of pro-apoptotic bax. In short, the results here showed that ZEA possessed comparative estrogenic activity and could promote the progression of MCF-7 cells through the cell cycle by a decrease in G(0)/G(1) phase and a significant increase in S phase. The pro-proliferative activity of ZEA was due to inhibition of apoptosis through regulation of bax/bcl-2 expression. Therefore, we conclude that contamination of ZEA in food might contribute to the increasing incidence rates of breast cancer. PMID- 16216641 TI - Lessons from severe acute respiratory syndrome (SARS): implications for infection control. AB - Severe acute respiratory syndrome (SARS), the first global epidemic in the 21st century, affected over 8500 people in approximately 30 countries . With a crude mortality of 9%, its cause was quickly identified as a novel coronavirus that jumped species from animals to man. The SARS coronavirus epidemic, which began in the Fall of 2002, was related to the exotic food industry in southern China, initially involving disproportionate numbers of animal handlers, chefs, and caterers. Subsequently, person-to-person transmission spawned the outbreak. What distinguished this illness clinically was the fact that approximately half of the victims were health care workers , infected while caring for recognized or unrecognized patients with SARS. There are many curiosities and uncertainties surrounding the epidemic of SARS with lessons that may be useful to the community of infectious diseases physicians, especially when looking ahead to the next epidemic. Herein we relate our perspectives on useful lessons derived from a review of the SARS epidemic. PMID- 16216642 TI - Long time due: reducing tuberculosis mortality in the 21st century. AB - The beginning and the end of the 20th century were marked by great pandemics: influenza and AIDS. Medical journals do not describe any major tuberculosis (TB) pandemics in the 20th century. Yet TB likely was responsible for more deaths in the last 100 years than influenza and HIV combined. Steadily, insidiously, millions of people die from TB every year. Even under optimal TB control conditions, it is estimated that more than 50 million people will die from TB between 1998 and 2020. Under current TB control conditions, the number is closer to 70 million. It is long past time that the global community committed to a serious program to eliminate tuberculosis mortality. Such a program would require making treatment universally available, making prevention accessible to those in poor countries as well as affluent, addressing the interaction between HIV and TB, and setting serious verifiable goals. A global 5 x 7 initiative that calls for treating an additional 5 million active TB cases per year, and for screening up to five contacts of every TB case, by 2007 would offer an important beginning. With the sustained effort that comes from public commitment, TB can be changed from one of the most important causes of preventable death worldwide to a historical cause of death. Without this effort, TB will remain the silent, steady killer it has been for centuries. The rationale for action, potential and need for success are detailed in this article. PMID- 16216643 TI - Prions: who should worry about them? AB - Prion diseases, also called transmissible spongiform encephalopathies (TSEs), are a family of neurodegenerative disorders affecting both humans and animals. They are caused by the accumulation of an abnormal form of a protein known as prion that results in neuronal death and a characteristic spongiform appearance of the brain tissue. Human prion diseases can be sporadic, acquired or hereditary. Acquired prion diseases have been linked to entering contaminated food into the human food chain, failure to completely disinfect or sterilize contaminated surgical instruments, patients receiving tissues and organs from infected donors, recipients of blood and other biological contaminated products, and potentially to cross infection in dental procedures. At present, there is unfortunately no efficient therapy that can be administered to clinically infected patients with prion diseases. Moreover, there are no simple diagnostic tests that can be used to show the agent of transmissible spongiform encephalopathy during the preclinical phase of the disease. Therefore, to prevent the spread of this emerging infectious agent it is necessary to implement several health control strategies and maintain surveillance for subclinical infections. PMID- 16216644 TI - Influenza: forecast for a pandemic. AB - Recently, great speculation about a possible influenza pandemic has been made. However, the facts supporting the possibility of this threat are less discussed. During the last decade highly pathogenic strains of avian influenza virus, including the H5N1 subtype, crossed the species barriers from birds to humans and caused fatal disease. The Z strain of H5N1 subtype is characterized by pathogenicity to a larger number of animal species and by resistance to the older class of antiviral drugs. At present, two out of three general conditions for the onset of a pandemic have been met; namely, the emergence of a new virus and its ability to replicate in humans causing serious illness. Should the virus achieve efficient human-to-human transmission, the next influenza pandemic might occur. This review addresses these biological and epidemiological aspects of influenza in the context of history and characteristics of previous epidemics, as well as concrete actions that can be undertaken considering current understanding of influenza pathogenesis, treatment, and control possibilities. PMID- 16216645 TI - Infection control: old problems and new challenges. AB - Infection control faces radical changes at the beginning of the third millennium. The first part of this review focuses on problems not yet solved, such as 1) surveillance systems, which should be active and extremely flexible; 2) infection outbreaks in hospitals and strategies to avoid them; 3) hand washing and alternatives such as rapid hand antisepsis; 4) water and food in the hospital as potential reservoirs of nosocomial pathogens; 5) upgrading of infection control programs to turn them into systems to improve the quality of care; 6) fatal Gram negative bacteremias in hospitals from developing countries, which can be avoided with better standards of care; 7) the elemental role of the microbiology laboratory in the prevention and control of infections; 8) the unprecedented crisis due to the emergence of specific multi-resistant pathogens; 9) the risks for healthcare workers, such as tuberculosis, hepatitis, HIV, SARS, and hemorrhagic fevers; and 10) the need for the consistent application of guidelines. The second part of this review focuses on new challenges for infection control, such as 1) the ever-growing number of immunocompromised patients and basic control measures to avoid opportunistic infections; 2) the concerns about the capacity of the public health systems to deal with terrorist acts; 3) the practice of high-risk procedures in facilities lacking trained personnel, efficient laboratories, and protective items; and 4) gene therapy and its potential infectious complications. Consideration is given to the asymmetric development of infection control globally. PMID- 16216646 TI - Bacteremias: a leading cause of death. AB - Bloodstream infections (BSIs), recognized to be a major cause of morbidity and mortality globally, are increasing in incidence. The reported rates of crude and attributable mortality vary, possibly due to heterogeneity in patient populations and methodology. Few studies, however, have focused on pathogen-specific attributable mortality. These studies include S. aureus, coagulase-negative staphylococci and enterococcus. Other studies of attributable mortality have been conducted in select populations such as nosocomial and community-acquired cohorts, intensive care units, neutropenic patients, and HIV-positive patients. Regrettably, despite advances in treatment and intensive care facilities, mortality remains high. PMID- 16216647 TI - Invasive candidiasis in immunocompromised hospitalized patients. AB - The frequency of infections by Candida species is increasing worldwide, with candidemia representing the fourth most common bloodstream infection in the U.S. The risk of infection is especially high in the immunocompromised, hospitalized patient. The treatment of and prophylaxis for Candida infection have led to the emergence of resistant species and the acquisition of resistance in previously susceptible species. Current therapeutic options include amphotericin B and its lipid compounds, fluconazole, itraconazole, voriconazole, and caspofungin. Research is focusing on better diagnostics and the evaluation of strategies such as prophylaxis in high-risk hosts and pre-emptive therapy. PMID- 16216648 TI - Sepsis: still going strong. AB - Sepsis definitions help to understand and to better define a group of syndromes secondary to an infectious insult. The hierarchical continuum of inflammatory response leads, in absence of counterregulatory forces, to organ damage and death. We have learned first the response to treatment and afterwards the pathophysiology behind it. This lesson has, of course, not always been followed by a reduction of mortality. The definition, natural history, risk factors, diagnoses, and treatment based on emerging evidence will help to improve patient outcomes and mortality. Standardized care seems to improve survival, and validation and further evaluation of this care is necessary to maximize resources and outcomes. PMID- 16216649 TI - AIDS: the second wave. AB - Over two decades ago, the first cases of a new disease now known as AIDS were described in the literature. Since then, remarkable advances have been made, a new retrovirus has been discovered, promising antiviral agents have been developed and affordable and effective preventive interventions exists. Yet the AIDS pandemic continues largely unabated. AIDS is the world's leading infectious disease cause of death and threatens to devastate the economic systems of resource-poor nations. Over the next decade the HIV epidemic is expected to continue to grow with an increasing proportion of infected persons residing in developing countries and increasingly affecting young women. However, reducing the burden of HIV/AIDS is achievable, and there is some cause for cautious optimism. PMID- 16216650 TI - Global warming and infectious disease. AB - Global warming has serious implications for all aspects of human life, including infectious diseases. The effect of global warming depends on the complex interaction between the human host population and the causative infectious agent. From the human standpoint, changes in the environment may trigger human migration, causing disease patterns to shift. Crop failures and famine may reduce host resistance to infections. Disease transmission may be enhanced through the scarcity and contamination of potable water sources. Importantly, significant economic and political stresses may damage the existing public health infrastructure, leaving mankind poorly prepared for unexpected epidemics. Global warming will certainly affect the abundance and distribution of disease vectors. Altitudes that are currently too cool to sustain vectors will become more conducive to them. Some vector populations may expand into new geographic areas, whereas others may disappear. Malaria, dengue, plague, and viruses causing encephalitic syndromes are among the many vector-borne diseases likely to be affected. Some models suggest that vector-borne diseases will become more common as the earth warms, although caution is needed in interpreting these predictions. Clearly, global warming will cause changes in the epidemiology of infectious diseases. The ability of mankind to react or adapt is dependent upon the magnitude and speed of the change. The outcome will also depend on our ability to recognize epidemics early, to contain them effectively, to provide appropriate treatment, and to commit resources to prevention and research. PMID- 16216651 TI - Resistance to antibiotics: are we in the post-antibiotic era? AB - Serious infections caused by bacteria that have become resistant to commonly used antibiotics have become a major global healthcare problem in the 21st century. They not only are more severe and require longer and more complex treatments, but they are also significantly more expensive to diagnose and to treat. Antibiotic resistance, initially a problem of the hospital setting associated with an increased number of hospital-acquired infections usually in critically ill and immunosuppressed patients, has now extended into the community causing severe infections difficult to diagnose and treat. The molecular mechanisms by which bacteria have become resistant to antibiotics are diverse and complex. Bacteria have developed resistance to all different classes of antibiotics discovered to date. The most frequent type of resistance is acquired and transmitted horizontally via the conjugation of a plasmid. In recent times new mechanisms of resistance have resulted in the simultaneous development of resistance to several antibiotic classes creating very dangerous multidrug-resistant (MDR) bacterial strains, some also known as "superbugs". The indiscriminate and inappropriate use of antibiotics in outpatient clinics, hospitalized patients and in the food industry is the single largest factor leading to antibiotic resistance. In recent years, the number of new antibiotics licensed for human use in different parts of the world has been lower than in the recent past. In addition, there has been less innovation in the field of antimicrobial discovery research and development. The pharmaceutical industry, large academic institutions or the government are not investing the necessary resources to produce the next generation of newer safe and effective antimicrobial drugs. In many cases, large pharmaceutical companies have terminated their anti-infective research programs altogether due to economic reasons. The potential negative consequences of all these events are relevant because they put society at risk for the spread of potentially serious MDR bacterial infections. PMID- 16216652 TI - The initial hospital response to an epidemic. AB - The last decades have been characterized by the appearance of a substantial number of newly recognized or novel infectious agents and by the re-emergence of infectious diseases with a global impact. The objective of this article is to briefly describe the model of hospital response for early diagnosis and prompt management of patients with highly contagious infectious diseases. We reviewed the main components of hospital preparedness in response to clustering of highly contagious diseases. A model for the initial hospital response to an epidemic in our referral Institute is discussed. Prompt recognition and identification is the initial and indispensable step in facing any communicable diseases, regardless of whether it is a prevalent, a newly emerging one or deliberately released. The importance of developing and implementing nontraditional methods of public health surveillance and a system that allows a wide and immediate dissemination of information and exchange of views on risk assessment and risk management are highlighted. Case identification and laboratory capabilities and isolation procedures are the essential components for an initial hospital response. The recent bioterrorist events and the worldwide outbreaks of highly contagious infectious diseases have evidenced the need for institutional preparedness at each hospital and for identification of referral centers for patient isolation and of laboratories with adequate capabilities. Moreover, hospitals should develop a plan for coordinating all hospital components to respond to critical situations deriving from the admission of patients with highly contagious infectious diseases. PMID- 16216653 TI - The modern autopsy: what to do if infection is suspected. AB - Deaths due to infectious diseases are common worldwide. The autopsy, although less frequently performed than previously, is important to our understanding of disease pathogenesis. The autopsy also provides critical information regarding potential disease outbreaks. To optimize the benefits of an autopsy, the pathologist should approach the autopsy with a well-constructed differential diagnosis that provides the framework for appropriate selection of diagnostic specimens and tests. Standard microbiologic cultures, although necessary and important, are often insufficient and must be supplemented by newer molecular methodologies. PMID- 16216654 TI - The internet and the global monitoring of emerging diseases: lessons from the first 10 years of ProMED-mail. AB - Now in its eleventh year, ProMED-mail is a robust and sensitive mechanism for the discovery of emerging disease outbreaks involving humans, animals, and plants around the world and for rapid dissemination of information. Fast reporting translates into quicker arrival of prevention and control assistance from other countries, better advance warning to neighboring countries and intending travelers, and other benefits to public health such as heightened awareness of health workers to the threat, time to prepare countermeasures and to react in an appropriate manner rather than a reflex emergency response. PMID- 16216655 TI - The contribution of international agencies to the control of communicable diseases. AB - Although inequality is often measured through three critical indicators education, income and life expectancy-health-related differences are also essential elements for explaining levels of equality or inequality in modern societies. Investment and investigation in health also involve inequalities at the global level, and this includes insufficient North-South transfer of funds, technology and expertise in the health field, including the specific area of communicable diseases. Globally, epidemics and outbreaks in any geographic region can represent international public health emergencies, and this type of threat requires a global response. Therefore, given the need to strengthen the global capacity for dealing with threats of infectious diseases, a framework is needed for collaboration on alerting the world to epidemics and responding to public health emergencies. This is necessary to guarantee a high level of security against the dissemination of communicable diseases in an ever more globalized world. In response to these needs, international health agencies have put a number of strategies into practice in order to contribute to the control of communicable diseases in poor countries. The principle strategies include: 1) implementation of mechanisms for international epidemiologic surveillance; 2) use of international law to support the control of communicable diseases; 3) international cooperation on health matters; 4) strategies to strengthen primary care services and health systems in general; 5) promotion of the transfer of resources for research and development from the North to the South. PMID- 16216656 TI - Use of computerized surveillance to detect nosocomial pneumonia in neonatal intensive care unit patients. AB - BACKGROUND: Pneumonia surveillance is difficult and time-consuming. The definition is complicated, and there are many opportunities for subjectivity in determining infection status. OBJECTIVE: To compare traditional infection control professional (ICP) surveillance for pneumonia among neonatal intensive care unit (NICU) patients with computerized surveillance of chest x-ray reports using an automated detection system based on a natural language processor. METHODS: This system evaluated chest x-rays from 2 NICUs over a 2-year period. It flagged x rays indicative of pneumonia according to rules derived from the National Nosocomial Infection Surveillance System definition as applied to radiology reports. Data from the automated system were compared with pneumonia data collected prospectively by an ICP. RESULTS: Sensitivity of the computerized surveillance in NICU 1 was 71%, and specificity was 99.8%. The positive predictive value was 7.9%, and the negative predictive value (NPV) was >99%. Data from NICU 2 were incomplete. CONCLUSIONS: Computer-assisted surveillance has the potential to decrease ICP workload and make pneumonia surveillance feasible. The high NPV means the system can safely screen out many chest x-rays of noninfected patients. However, all data must be available to the computer system and must be analyzed the same way for results to be comparable. PMID- 16216657 TI - Handheld computer surveillance: shoe-leather epidemiology in the "palm" of your hand. AB - BACKGROUND: Although urinary tract infections (UTIs) are the most common nosocomial infection, active UTI surveillance is often not feasible for infection control departments. METHODS: As part of an ongoing urinary catheter evaluation, we investigated the accuracy and cost-effectiveness of using handheld personal digital assistants (PDA) and computer-based UTI surveillance in comparison with traditional surveillance of UTIs among medical intensive care unit (MICU) patients. From September 22, 2000, to October 22, 2000, an infection control practitioner (ICP) actively surveyed all MICU patients who had a urinary catheter to determine criteria for a nosocomial UTI as defined by the Centers for Disease Control and Prevention (CDC). Research assistants visited the unit each day to determine which patients had urinary catheters, and this information was uploaded to a database designed for laboratory-based UTI surveillance. Patients with a urinary catheter were included if admitted to the MICU for >48 hours and excluded if symptoms developed <48 hours after admission. RESULTS: Both data collection methods identified 8 UTIs within the 1-month period, with 3 UTIs meeting inclusion criteria. Traditional surveillance required 8 hours weekly, whereas the PDA-computer surveillance required 1-and-a-half hours weekly. Traditional surveillance correctly attributed all 3 UTIs to the MICU. The PDA-computer surveillance incorrectly attributed the location of 2 of the UTIs. In both cases, the UTI occurred less than 48 hours after the patient was transferred from the MICU. CONCLUSION: Our data demonstrate that PDA surveillance detects nosocomial UTIs efficiently and is a novel, timesaving technique that realizes a cost savings in the first year. PMID- 16216658 TI - Surgical site infection rates following cardiac surgery: the impact of a 6-year infection control program. AB - BACKGROUND: To evaluate the impact of an infection control program on surgical site infections (SSIs) complicating cardiac operations. METHODS: Prospective cohort study of patients undergoing cardiac operations. Interventions included prospective surveillance, povidone-iodine scrub showers, depilation before surgery, administration of preoperative antibiotic prophylaxis in the operating room, and postdischarge follow-up. Logistic regression models were fitted to assess infection rates over time, adjusting for factors known to affect SSI rates. RESULTS: The overall SSI rate for 2051 procedures was 10.4%. Rates of superficial and deep incisional SSIs remained unchanged over the study period. The rates of all organ/space infections, mediastinitis, and SSIs because of methicillin-resistant Staphylococcus aureus during the first 2 years were 3.25%, 2.22%, and 1.48%, respectively, and they decreased to 1.17%, 0.73%, and 0.73%, respectively, by the end of 2002 (P = .01, P = .01, and P = .09, respectively). The adjusted odds ratios for these 3 types of infection at the end of 2002 compared with December 31, 1998, were 0.19 (95% confidence interval [95% CI]: 0.07-0.48), 0.20 (95% CI: 0.06-0.66), and 0.28 (95% CI: 0.08-0.97), respectively. CONCLUSION: We observed significant reductions in organ/space infection rates, particularly mediastinitis. These differences remained significant when adjusted for potential confounding variables. PMID- 16216659 TI - Evaluation of postdischarge surveillance of surgical site infections after total hip and knee arthroplasty. AB - BACKGROUND: Artificial joint replacement of hip (HPRO) and knee (KPRO) are 2 of about 20 categories of operative procedures of the surveillance of surgical site infection (SSI) as stated by nosocomial infections surveillance systems in the United States and in Germany. Periprosthetic SSI can manifest itself after a long period. METHODS: Seven hundred fifty-six orthopedic patients from 2 centers were evaluated after HPRO (n = 508) or KPRO (n = 248). SSI was recorded during hospitalization and for 12 month postdischarge. The surveillance regimen was extended by also sending patients a questionnaire after 12 months postdischarge. All complaints were followed up by contacting the patients and any clinicians and general practitioners (GPs) involved. Stratified infection rates and standardized infection ratio (SIR) were calculated and compared with reference data of the national surveillance system. RESULTS: The total response rate to the postal questionnaire survey was 85.2%. SSI was recorded in 16 patients (3.15%) after HPRO; 12 were detected by predischarge surveillance, and the 4 cases found postdischarge were all organ/space SSI. In total, only 1 SSI was detected after KPRO before discharge and none after discharge (SSI rate 0.40%). Time between discharge and detection of SSI cases ranged from 8 days to 8 months. SIR of HPRO was 1.25 and SIR of KPRO was 0.36. CONCLUSION: Because 25% of SSIs after HPRO occurred after discharge and all were organ/space SSI, highlights the importance of postdischarge surveillance of nosocomial infections (NIs). Because all SSIs were reported already by current surveillance, the extended postdischarge surveillance appears to be unnecessary. The pursuit of shorter hospital stay after surgery may challenge the methods of surveillance systems in future. PMID- 16216660 TI - Policies to increase influenza and pneumococcal immunizations in chronically ill and institutionalized settings. AB - BACKGROUND: The objective of this study was to understand better the status of and ways to improve dissemination of influenza and pneumococcal standing-order vaccination policies to at-risk adults in health care institutions. METHODS: A statewide sample of 5 different types of institutions serving at-risk elderly persons in North Carolina was surveyed. A 45-question telephone survey was administered to infection control nurses or facility directors at 267 (86% response rate) health care facilities involved in direct patient care. RESULTS: A majority of respondents reported that influenza (81%) and pneumococcal (59%) diseases were important to their facility, and 63% stated that the influenza vaccine was very effective versus 47% for pneumococcal. Except nursing homes, few facilities reported adoption of standing-order policies to vaccinate routinely the at-risk adults. Over 70% of respondents stated that their facilities might consider adopting standing-order policies for influenza and pneumococcal disease. A majority of respondents also supported a state law that requires such vaccines for high-risk patients unless contraindicated or the patient refuses. CONCLUSIONS: Respondents across diverse health care institutions appear interested in adopting standing-order policies to increase influenza and pneumococcal vaccination rates and are more likely to do so if provided with appropriate administrative and/or financial support for implementation. PMID- 16216661 TI - Specific risk factors for Clostridium difficile-associated diarrhea: a prospective, multicenter, case control evaluation. AB - BACKGROUND: Clostridium difficile is a toxin-producing bacterium that is responsible for toxicity to the colonic mucosa, causing inflammation, necrosis, and, in some extreme cases, intestinal dilation and perforation. C difficile associated diarrhea (CDAD) occurs when patients have a reduction in their natural gastrointestinal flora that allows for the proliferation of and toxin production by C difficile. METHODS: Using a multicenter, prospective observational case control study, we assessed and quantified risk factors associated with the development of diarrhea caused by Clostridium difficile, with particular attention to antibiotic use. All hospitalized patients with diarrhea requiring a C difficile toxin test as part of their routine clinical workup were considered for study inclusion. Patients with a negative specimen (controls) were considered for enrollment if matched (by age, sex, length of stay, and institution) to a case. Variables associated with CDAD were identified using univariate analysis. Significant factors were then entered into multivariate logistic regression analysis to identify independent factors. RESULTS: There were no significant differences in antibiotic use between cases and controls. Patient severity, classified by Horn's Index, was significantly different between cases and controls (P = .0022). No other significant variables were identified. CONCLUSION: The severity of illness of the cases was classified as more severe than the controls, but no significant differences in antibiotic use were identified between the groups. The negative C difficile toxin studies on the well-matched control patients indicate a different etiology of diarrhea (such as antibiotic associated diarrhea), which may have developed in the presence of similar antibiotic use as the cases. PMID- 16216662 TI - Influenza immunization of medical residents: knowledge, attitudes, and behaviors. AB - BACKGROUND: There have been few studies of barriers to acceptance of influenza immunization among medical residents. EXPERIMENTAL DESIGN: We conducted a cross sectional survey of residents at the Western Pennsylvania Hospital during the 2003-2004 influenza season. An anonymous questionnaire designed specifically for this study was used to collect demographic, health beliefs and attitudes, and medical knowledge data related to the influenza vaccine. RESULTS: 43 residents were surveyed from January to February 2004. 58% of the respondents reported receiving the vaccine. Immunization rates were significantly associated with postgraduate level, prior vaccination, media influence, whether they knew co residents who were vaccinated, medical knowledge scores, and plan to be vaccinated next year. Immunization rates by age, sex, type of medical school, department, whether they had children younger than 16, whether they would recommend the vaccine to patients, and the respondents' health status did not differ significantly. Residents who had higher medical knowledge scores were significantly more likely to be immunized and recommend the vaccine to patients. CONCLUSIONS: Resident influenza immunization rate in this sample was higher than the national average for healthcare workers. The rate of immunization was associated with demographic, knowledge, and behavioral factors. PMID- 16216663 TI - Clean care is safer care: the first global challenge of the WHO World Alliance for Patient Safety. PMID- 16216664 TI - A statewide survey of nosocomial infection surveillance in acute care hospitals. AB - Increasingly, states are considering mandating the reporting of nosocomial infection data. To determine the impact of potential legislation, a questionnaire was mailed to the infection control department of each hospital in Virginia to assess the size of the infection control workforce and methodologies used for nosocomial infection surveillance. Most hospitals (64%) had 1 ICP full-time equivalent (FTE), and, at 86% of hospitals, the ICPs had other major responsibilities. The estimated mean additional ICP FTE required to perform hospital-wide surveillance was 1.7. Statewide, an additional 160 ICPs at an estimated annual cost of 11.5 million dollars would be required if reporting of all nosocomial infections were mandated. PMID- 16216665 TI - Colonization of Yankauer suction catheters with pathogenic organisms. AB - Oral suction devices may be fomites for nosocomial infections. This study was designed to evaluate the rate of contamination of Yankauer suction catheters. Among the 20 catheters tested, 16 (80%) yielded cultures for pathogens. Seven (35%) were colonized with multiple pathogens. Among the organisms encountered included methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant Enterococci (VRE). These devices should be handled and stored with appropriate care. PMID- 16216666 TI - Knowledge, attitudes, and practices of contact precautions among Iranian nurses. AB - BACKGROUND: Knowledge, attitudes, and contact precaution practices were surveyed in nurses at Shiraz University of Medical Sciences. METHODS: Two hundred seventy nurses, midwives, and auxiliary nurses completed a questionnaire consisting of 8 knowledge items with corresponding attitudes and practices items. RESULTS: Compliance with precaution practices was low, 19.5%, and little more than half, 51.8%, held positive attitudes toward the guidelines, whereas 65.5% could correctly answered all precaution knowledge items. Nurses with correct knowledge were 11.3 times more likely (P < .0001) to hold positive attitudes toward the guidelines and 14.2 times more likely (P < .0001) to comply with all 8 precaution practices, and compliers were 6.3 times more likely (P < .0001) to hold positive attitudes than noncompliers. CONCLUSION: Although correct knowledge was associated with compliance and positive attitudes, the proportion of nurses who held positive attitudes also had good knowledge, and compliance with practices was not abundant. Better training coverage may result in compliance with precaution practices becoming the norm. PMID- 16216668 TI - Report of the Wet Workshop for Quantification of Soluble HLA-G in Essen, 2004. AB - Membrane-anchored and soluble human leukocyte antigen HLA-G (sHLA-G) molecules exert strong inhibiting signals after interaction with their cognate receptors ILT2 (CD85j), ILT4 (CD85d), and KIR2DL4 (CD158d) that are differentially expressed by natural killer cells, T cells, and antigen-presenting cells. These inhibitory functions can become operative in conditions in which such immune cells try to attack viral infected or tumor cells. Recently, clinical studies showed that sHLA-G molecules are also relevant in the prediction of allograft acceptance after heart transplantation, liver-kidney cotransplantation, and the successful implantation and development of embryos after in vitro fertilization. In view of this diagnostic potential, reliable methods for the measurement of sHLA-G molecules in various body fluids are of interest. Thus, the aims of the Wet Workshop for measurement of sHLA-G held in Essen, Germany (at the Institute of Immunology October 18-20, 2004) were to select and validate HLA-G-specific enzyme-linked immunosorbent assay (ELISA) formats and purified standard HLA-G proteins, which can be easily generated and used as consensual references. To this end, the antibody combinations monoclonal antibody (mAb) MEM-G/9 (capture) + anti-beta2m (detection) and the mAb 5A6G7 (capture) + mAb W6/32 (detection) were chosen in an ELISA format for the simultaneous determination of shed HLA-G1 + soluble HLA-G5 (sHLA-G1 + HLA-G5) and for the exclusive detection of HLA-G5 molecules, respectively. As standard, protein HLA-G5 molecules were purified from insect SF9 cells coinfected by HLA-G5 + human beta2m and characterized for their antigenic determinants. A total of 24 members in 13 teams participated in the 3 day sHLA-G Wet Workshop. All workshop materials, protocols, standard reagents, and samples were provided to each team by the organizers. The Wet-Workshop results clearly demonstrated that (1) the HLA-G5 standard reagent was equally detected by both ELISA formats; (2) sHLA-G1 + G5 and HLA-G5 molecules, respectively, were specifically detected by the two ELISA formats; and (3) both ELISA formats measure reproducibly the amounts of sHLA-G. The comparison of the two ELISA results obtained evidenced that in healthy donors sHLA-G1 molecules can exist in body fluids besides HLA-G5. Moreover, a novel soluble HLA-G structure can be predicted that is recognized by the mAb 5A6G7 + mAb W6/32 antibody combination, but not by the one of mAb MEM-G/9 + anti-beta2m. PMID- 16216667 TI - Preventing infections in nursing homes: a survey of infection control practices in southeast Michigan. AB - BACKGROUND: Studies on adherence to infection control policies in nursing homes (NHs) are limited. This pilot study explores the use of various infection control practices and the role of infection control practitioners in southeast Michigan NHs. METHODS: A 43-item self-administered questionnaire and explanatory cover letter were mailed to 105 licensed NHs in southeast Michigan. A second mailing was sent to the nonresponders 4 weeks later. RESULTS: Significant variability existed in adoption of various infection control measures with respect to time spent in infection control activities (50% of facilities having a full-time infection control practitioner), definitions used in monitoring infections, and immunization rates (influenza: range, 0%-100%; mean, 73.2%; pneumococcal: range, 0%-100%; mean, 38.5%). CONCLUSION: Although strides have been made in infection control research in NHs, significant variations exist in implementation of infection control methods and guidelines. Future research should focus on identifying barriers to infection control in NHs. PMID- 16216669 TI - Analysis of vascular endothelial growth factor (VEGF) functional variants in rheumatoid arthritis. AB - The vascular endothelial growth factor (VEGF) is one of the most important pro angiogenic mediators related to inflammation-associated synovial angiogenesis. The aim of this study was to asses the role of -1154 G-->A (rs1570360) and -634 G ->C (rs2010963) VEGF gene functional variants with rheumatoid arthritis (RA). The population under study was composed of a total of 753 unrelated RA patients and 801 healthy controls. The VEGF -1154 G-->A and -634 G-->C polymorphism genotyping was performed by real-time polymerase chain reaction technology, using TaqMan 5' allelic discrimination assay. No evidence of association was observed between the -1154 G-->A and the -634 G-->C VEGF polymorphisms, or inferred VEGF haplotypes with RA susceptibility or clinical manifestations. Our results suggest that the analyzed VEGF promoter polymorphisms may not play a relevant role in RA pathogenesis in our population. PMID- 16216671 TI - MHC class I-associated peptides identified from normal platelets and from individuals with idiopathic thrombocytopenic purpura. AB - Major histocompatibility complex (MHC) class I molecules bind and display peptide antigens on the cell surface. CD8(+) T lymphocytes recognize peptides in association with class I proteins to initiate a cytotoxic immune response. To understand the specificity of such immune responses and to facilitate the development of therapies for disease, it is important to identify MHC-presented peptides. In this study, platelets, easily obtainable and often associated with immune-mediated disease, were selected to identify MHC class I-associated peptides. MHC-associated peptides presented on platelets of normal individuals and individuals with idiopathic thrombocytopenic purpura (ITP) were characterized. ITP is characterized by the premature immune destruction of platelets. It is associated with the production of antiplatelet autoantibodies, most often targeting platelet membrane GPIIb/IIIa or GPIb/IX. In addition to characterizing five fully and several partially sequenced peptides from platelets, the peptide GPRGA(L/I)S(L/I)(L/I) was identified from four of the five ITP patients. The anchor motif of this peptide correlates with the presence of the HLA-B7 allele. A BLAST search identified this peptide as GPIb (4-12). In conclusion, platelets from normal and ITP individuals can present peptides from general cellular proteins and platelet specific proteins, such as GPIb, to the immune system via MHC class I. PMID- 16216670 TI - Analysis of a GT microsatellite in the promoter of the foxp3/scurfin gene in autoimmune diseases. AB - The aim of this study was to assess the possible association of the functional (GT)(n) microsatellite polymorphism in the FOXP3 gene with predisposition to several autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), ulcerative colitis (UC), Crohn's disease, and celiac disease. We analyzed a case-control cohort composed of 231 SLE patients, 293 RA patients, 528 inflammatory bowel disease (354 Crohn's disease patients and 260 UC patients) patients, 103 celiac disease patients, and 274 healthy controls ethnically matched. Genotyping of (GT)(n) microsatellite was performed by polymerase chain reaction (PCR)-based method combined with fluorescent technology. We found no evidence for association of this polymorphism between controls and these autoimmune disease patients. Additionally, no differences in the genotype and allele distribution were found when patients were stratified according to clinical manifestation. The (GT)(n) microsatellite of the FOXP3 gene may not play a relevant role in the susceptibility to SLE, RA, inflammatory bowel disease, and celiac disease in our population. PMID- 16216672 TI - Continuous presence of Th1 conditions is necessary for longer lasting tumor specific CTL activity in stimulation cultures with PBL. AB - The generation of tumor-associated, but self-antigen specific cytotoxic T lymphocytes (CTL) response is possible by vaccination even in patients at the advanced stages of the disease. The in vivo expansion of such CTLs is now the most important objective of the immunotherapy. In human melanoma, we show here that MART-1(27-35)-specific CTLs generated with purified CD8+ cells survive and maintain their activity longer in culture than those CTLs generated by using total peripheral blood lymphocytes (PBL) taken either from patients or from normal donors. When PBL are grown under Th1 conditioning the quantity and quality of CTL with total PBL are comparable with that of the CTLs generated with purified CD8+ cells. For patients either autologous melanoma tumor cells or MART 1(27-35) peptide pulsed autologous DC were used to generate CTL responses. For normal donors MART-1(27-35) peptide pulsed autologous DC were used. For both normal donors or patients, polarization of PBL with Th1 conditioning with interleukin (IL)-12 (250 U/ml) and anti IL-4 antibody 1 mug/ml for 7 days before CTL generation, induced better and longer living CTL response and prevented the expansion of CD4+ T cells that have downregulatory activity. We show that continuous presence of Th1 conditioning in cultures with total PBL generated significantly higher number of antigen-specific CTLs as detected by MART-1 HLA-A2 tetramer staining. The antigen specificity of such CTLs were determined by IFN gamma secretion in response to target cells bearing the specific antigen. Our observations indicate that Th1 conditioning results in a longer lasting CTL response in vitro and points toward a newer approach for vaccine strategy. PMID- 16216673 TI - Characterization of mannose-binding lectin gene polymorphism among human T-cell lymphotropic virus 1 and 2-infected asymptomatic subjects. AB - The present study investigated the association between mannose-binding lectin (MBL) gene polymorphism and the susceptibility to human T-cell lymphotropic virus (HTLV) infection in a group of 83 HTLV-infected asymptomatic subjects (62 HTLV-1 and 21 HTLV-2) and 99 healthy controls. Detection of MBL*A, MBL*B, and MBL*C was performed by amplifying a fragment of 349 bp (exon 1) and submitting the product to restriction fragment length polymorphism analysis with BanI and MboII endonucleases. Allele MBL*D was investigated by sequence-specific primer polymerase chain reaction. The frequency of MBL*A, MBL*B, and MBL*D was 63%, 22%, and 15% among seropositive subjects and 70%, 14%, and 16% among healthy controls, respectively. Genotype differences were statistically significant (chi2 = 11.57; p = 0.04); the presence of genotype BB was 9.6% among HTLV-infected patients compared with 1% among controls (chi2 = 7.151; p = 0.019). A significant difference of the genotype frequencies between HTLV-1 and HTLV-2 infections was observed, but this result could be attributed to the number of investigated HTLV 1-infected subjects. The odds ratio to the presence of BB genotype was 10.453 (1.279 < or = IC95% < or = 85.40; p = 0.019). Results reveal a strong association between MBL polymorphism and HTLV infection. Presence of genotype BB may be associated with the susceptibility to HTLV, but further studies, with a larger number of individuals, will be necessary. MBL polymorphism could possibly have an impact on diseases associated with HTLV infection. PMID- 16216674 TI - Th1 cytokine polymorphisms in spanish patients with type 1 diabetes. AB - Several polymorphisms in regions where Th1 cytokines (IL12B and IFNG genes) are located were analyzed in 303 Spanish subjects with type 1 diabetes and compared with a control cohort (n = 548). Both groups comprised residents of the Madrid area. The haplotype frequencies were estimated by the expectation-maximization algorithm, and p values were corrected by the number of haplotypes taken into account in the study. Two haplotypes were significantly associated with the disease, one in the IL12B region (D5S2038*8/D5S1352*2/SNP1188C; OR = 3.01, p(c) = 0.0255) and another involved the IFNGgene (D12S313*9/IFNG*1; OR = 1.58, p(c) = 0.0217). Furthermore, a protective IL12B haplotype was found (D5S2038*4/D5S1352*1/SNP1188A; OR = 0.40, p(c) = 0.0405). No association was found for any of IL12B and IFNG markers individually. PMID- 16216675 TI - HLA-DQA1 introns 2 and 3 sequencing: DQA1 sequencing-based typing and characterization of a highly polymorphic microsatellite at intron 3 of DQA1*0505. AB - DQA1 class II gene encodes the alpha-chain of the human leukocyte antigen (HLA) DQ heterodimer. Sequencing-based typing (SBT) for HLA genes is the most powerful methodology described. However, most of the SBT procedures reported for HLA class II genes are not able to define complete exon 2 region. For that purpose, we have characterized introns 2 and 3 from most DQA1 alleles to design amplification procedures that were able to obtain complete exon 2 and 3 sequences from DQA1 genes. This coding information allowed us to reduce the number of ambiguities for DQA1 typing. DQA1 intron 2 and 3 characterization demonstrated the presence of two polymorphisms for alleles with the same exons 2 and 3 sequence from DQA1*05 group. Different samples including the DQA1*050101 alleles showed a single nucleotide polymorphism at position 53 of intron 2 (G53T). Additional haplotypic analysis showed the possible association of T53 allele with the Ax-Cw5-B18-DR17 DQ2 extended haplotype. On the other hand, DQA1*0505 sequencing from different control samples noticed the existence of a microsatellite (TTTC/AAAG)n located at position 126 of intron 3. Fragment length analysis demonstrated a high polymorphism for this short tandem repeat system (0505STR), defining alleles that ranged from 8 to 20 repetitions in our population. PMID- 16216676 TI - Identification of HLA-A*0111N: a synonymous substitution, introducing an alternative splice site in exon 3, silenced the expression of an HLA-A allele. AB - A new variant of the HLA-A*010101 allele designated as HLA-A*0111N, previously known as HLA-A*010101var, was identified in a patient requiring a stem-cell transplantation. The patient was typed by serologic methods as HLA-A2 homozygous and by sequence-based typing (SBT) as A*010101,020601. Flow-cytometric (FCM) analysis with 11 human monoclonal antibodies (mAbs) for the A1 molecule confirmed lack of any cell membrane expression of the A*0111N allele. One-dimensional isoelectric focusing (1D-IEF) of total cell lysate from the patient's cells revealed no cell surface and cytoplasmic A1 protein expression, whereas the HLA A2 molecule was identified by both FCM analysis and 1D-IEF. DNA sequence analysis showed the presence of a synonymous substitution from G to T at position 597 in codon 175. RNA SBT revealed a deletion of 24 bp in exon 3, position 596 through 619, encoding codons 175 through 182 of the HLA-A*0111N allele. The synonymous substitution introduced a new splice site, resulting in an efficient splicing, because no classical A1 protein could be detected in the patient. This alternative splicing prevented the translation into a correct and stable class I molecule expression on the cell surface. PMID- 16216677 TI - TAP1 and TAP2 polymorphisms and their linkage disequilibrium with HLA-DR, -DP, and -DQ in an eastern Andalusian population. AB - Transporter associated with antigen processing (TAP) molecules are involved in the processing of endogenous peptides that bind to major histocompatibility complex (MHC) class I molecules. The possible functional significance of TAP polymorphisms for antigenic peptide transport is an unresolved issue. Population genetics is a tool for investigating the evolutionary and functional significance of genetic polymorphisms. We studied 105 unrelated individuals from Eastern Andalusia in Southern Spain for TAP1 and TAP2 polymorphisms and to detect linkage disequilibrium between TAP1 and TAP2 and between TAP1/TAP2 and human lymphocyte antigen (HLA) DR, DP, and DQ genes. HLA-DR, -DQ, -DP, and TAP1 loci were genotyped with the polymerase chain reaction (PCR)-sequence-specific oligonucleotide method, and TAP2 genes were typed by using the amplification refractory mutation system-PCR technique. The alleles TAP1*D (3.3%), TAP2*D (2.4%), and TAP2*E (2.9%) were present in the Eastern Andalusian population but not in the general Spanish population. No evidence of linkage disequilibrium was found between TAP1 and TAP2 or between the TAP genes and HLA-DR, -DP, and -DQ in the Eastern Andalusian population. These results are consistent with the absence of coevolution between TAP and MHC class II genes and the hypothesis of selective neutrality. PMID- 16216678 TI - MICA polymorphism in a population from north Morocco, Metalsa Berbers, using sequence-based typing. AB - The MICA gene encodes a family of nonclassical major histocompatibility complex class I molecules. Data on MICA polymorphism in different populations are still limited. In the present study, MICA allele frequencies (af) were assessed in 82 unrelated healthy individuals from a Moroccan Berber population named Metalsa (ME) by means of sequence-based typing of exons 2, 3, 4, and 5. In consideration of the linkage disequilibrium existing between MICA and human leukocyte antigen (HLA) class I alleles, MICA/HLA-B, MICA/HLA-Cw, and MICA/HLA-A haplotype frequencies (hf) were estimated. A wide allelic distribution including 16 different MICA alleles was found in ME. The most common MICA alleles were MICA*00801 (af = 0.268), *004 (0.232), *00902 (0.140), *00901 (0.085), and *00901 (0.073). The most common MICA/HLA-B haplotypes were MICA*004-B*4403 and MICA*009 B*50 (hf = 0.113 for both these haplotypes). Some known MICA and HLA-B associations were confirmed in this population. Noteworthy was the high frequency of MICA*009 (af = 0.226); the high frequency of B*50 found in ME (af = 0.114) permitted us to evidence the associations of MICA*00902 with B*5001 (hf = 0.068) or *5002 (hf = 0.045), whereas MICA*00901 was mainly associated with B*5101 (hf = 0.038), which corresponds to the previously described association MICA*009/A6-HLA B*51. This study extends the previous knowledge on MICA polymorphism to a North African white population and may have implications for disease associations and transplantation. PMID- 16216679 TI - Proceedings of the 2nd Conference on NeuroEconomics. May 25-27, 2004. Munster, Germany. PMID- 16216680 TI - Neuroeconomics: an overview from an economic perspective. AB - Until now, economic theory has not systematically integrated the influence of emotions on decision-making. Since evidence from neuroscience suggests that decision-making as hypothesized in economic theory depends on prior emotional processing, interdisciplinary research under the label of "neuroeconomics" arose. The key idea of this approach is to employ recent neuroscientific methods in order to analyze economically relevant brain processes. This paper aims to provide an overview of the current state of neuroeconomic research by giving a brief description of the concept of neuroeconomics, outlining methods commonly used and describing current studies in this new research area. Finally, some future prospects and limitations are discussed. PMID- 16216681 TI - Neuroeconomics--from neural systems to economic behaviour. AB - Neuroeconomics is a new and highly interdisciplinary field. Drawing from theories and methodologies employed in both economics and neuroscience, it aims at understanding the neural systems supporting and affecting economically relevant behaviour in real-life situations. Although incomplete, the evidence is beginning to clarify with the possibility that neuroeconomic methodology might eventually trace whole processes of economically relevant behaviour. This paper accompanies the author's ConNEcs 2004 keynote speech on applications of neuroeconomic research. PMID- 16216682 TI - Experimental design in brain activation MRI: cautionary tales. AB - The use of functional magnetic resonance imaging (fMRI) in cognitive neuroscience has expanded at an amazing rate in the past 10 years. Current research includes increasingly subtle and specific attempts to dissect the cognitive and emotional mechanisms called into play when humans make decisions. The present essay will briefly review some of the general considerations and domains of information needed when one designs fMRI-based experiments. However, the main theme will be the difficulties associated with designing, conducting, analyzing and interpreting such research. Functional MRI is an unusually complicated technique, and there are numerous ways for experiments to go wrong. As well as demanding exceptional care in maintaining the quality of one's own research, this makes the universal problem of evaluating other peoples' research particularly challenging. PMID- 16216683 TI - Motivating forces of human actions. Neuroimaging reward and social interaction. AB - In neuroeconomics, reward and social interaction are central concepts to understand what motivates human behaviour. Both concepts are investigated in humans using neuroimaging methods. In this paper, we provide an overview about these results and discuss their relevance for economic behaviour. For reward it has been shown that a system exists in humans that is involved in predicting rewards and thus guides behaviour, involving a circuit including the striatum, the orbitofrontal cortex and the amygdala. Recent studies on social interaction revealed a mentalizing system representing the mental states of others. A central part of this system is the medial prefrontal cortex, in particular the anterior paracingulate cortex. The reward as well as the mentalizing system is engaged in economic decision-making. We will discuss implications of this study for neuromarketing as well as general implications of these results that may help to provide deeper insights into the motivating forces of human behaviour. PMID- 16216684 TI - Reward-based decision-making and aging. AB - Healthy aging is associated with a number of neuroanatomical and neurobiological alterations that result in various cognitive changes. Both, the dopaminergic as well as the serotonergic system are subject to change during aging. Receptor loss and severe structural changes in PFC and striatum have been reported. Aging is associated with a progressive decline in several cognitive functions, such as episodic memory, working memory, and processing speed. Furthermore, it is associated with deficits in tasks requiring adaptation to external feedback of right or wrong, or task-switching. Here, we develop the hypothesis that this loss of behavioral flexibility is caused by structural and functional alterations of the reward system leading to impairments in reward processing, learning stimulus reinforcement associations, and reward-based decision-making. We review (a) data on neural correlates and substrates of reward processing in young healthy animals and humans, (b) evidence for age related functional and structural alterations of the reward system, and (c) behavioral and neuroimaging data of age effects on reward-based decision-making processes. Implications for neuroeconomics and neurodegenerative diseases are discussed. PMID- 16216685 TI - The neuroscience of investing: fMRI of the reward system. AB - Functional magnetic resonance imaging (fMRI) has proven a useful tool for observing neural BOLD signal changes during complex cognitive and emotional tasks. Yet the meaning and applicability of the fMRI data being gathered is still largely unknown. The brain's reward system underlies the fundamental neural processes of goal evaluation, preference formation, positive motivation, and choice behavior. fMRI technology allows researchers to dynamically visualize reward system processes. Experimenters can then correlate reward system BOLD activations with experimental behavior from carefully controlled experiments. In the SPAN lab at Stanford University, directed by Brian Knutson Ph.D., researchers have been using financial tasks during fMRI scanning to correlate emotion, behavior, and cognition with the reward system's fundamental neural activations. One goal of the SPAN lab is the development of predictive models of behavior. In this paper we extrapolate our fMRI results toward understanding and predicting individual behavior in the uncertain and high-risk environment of the financial markets. The financial market price anomalies of "value versus glamour" and "momentum" may be real-world examples of reward system activation biasing collective behavior. On the individual level, the investor's bias of overconfidence may similarly be related to reward system activation. We attempt to understand selected "irrational" investor behaviors and anomalous financial market price patterns through correlations with findings from fMRI research of the reward system. PMID- 16216686 TI - Social memory, social stress, and economic behaviors. AB - Social memory plays a pivotal role in social behaviors, from mating behaviors to cooperative behaviors based on reciprocal altruism. More specifically, social/person recognition memory is supposed, by behavioral-economic and game theoretic analysis, to be required for tit-for-tat like cooperative behaviors to evolve under the N-person iterated prisoner's dilemma game condition. Meanwhile, humans are known to show a social stress response during face-to-face social interactions, which might affect economic behaviors. Furthermore, it is known that there are individual differences in a social stress response, which might be reflected in individual differences in various types of economic behaviors, partially via different capacities of social memory. In the present study, we investigated the acute effects of social stress-induced free cortisol (a stress hormone) elevation on hippocampus-dependent social memory by utilizing the Trier social stress test (consisting of a public speech and a mental arithmetic task). We also examine the correlation between an economic behavior-related personality trait (i.e., general trust scale) and social stress-induced cortisol elevations. We found that (1) social stress acutely impairs social memory during social interaction and (2) interpersonal trust reduces social stress response. Together, interpersonal trust may modulate economic behaviors via stress hormone's action on social cognition-related brain regions. PMID- 16216687 TI - Variants of uncertainty in decision-making and their neural correlates. AB - When leaving the tidy world of rules and people start judging probabilities on an intuitive basis, it revealed that they have some intuitions to choose from. One could refer to them as a family of subjective probability concepts or following Kahneman and Tversky, as variants of uncertainty. The authors distinguished between external and internal attributions of uncertainty and could show that the perceived reason of uncertainty determines the selected coping strategy. To investigate whether variants of uncertainty can also be distinguished on the cerebral level, two functional magnetic resonance imaging studies were conducted. Participants had to predict events (abstract visual stimuli) under parametrically varying degrees of (un-)certainty. In the first experiment, uncertainty was induced by the manipulation of event probability (externally attributed uncertainty). In the second experiment, uncertainty depended on participants' knowledge of valid rules of event occurrence, as trained before the experimental session (internally attributed uncertainty). As a result, parametric analyses revealed that activation within the posterior fronto-median cortex, particularly within mesial Brodmann area (BA) 8, increased with increasing uncertainty, no matter for which reason uncertainty emerged. Furthermore, it was found that different variants of uncertainty entailing different coping strategies can be dissociated due to additionally activated networks. Concluding, increasing activation within mesial BA 8 reflects that we are uncertain, additional networks what we do to resolve uncertainty in order to achieve future rewards. Hence, the phenomenological distinction between processes related to externally and internally attributed uncertainty is paralleled on the cerebral level. PMID- 16216688 TI - Evidence for a neural correlate of a framing effect: bias-specific activity in the ventromedial prefrontal cortex during credibility judgments. AB - BACKGROUND: Neural processes within the medial prefrontal cortex play a crucial role in assessing and integrating emotional and other implicit information during decision-making. Phylogenetically, it was important for the individual to assess the relevance of all kinds of environmental stimuli in order to adapt behavior in a flexible manner. Consequently, we can in principle not exclude that environmental information covertly influences the evaluation of actually decision relevant facts ("framing effect"). OBJECTIVE: To test the hypothesis that the medial prefrontal cortex is involved into a framing effect we employed functional magnetic resonance imaging (fMRI) during a binary credibility judgment task. METHODS: Twenty-one subjects were asked to judge 30 normalized news magazine headlines by forced answers as "true" or "false". To confound the judgments by formally irrelevant framing information we presented each of the headlines in four different news magazines characterized by varying credibility. For each subject the susceptibility to the judgment confounder (framing information) was assessed by magazine-specific modifications of the answers given. RESULTS: We could show that individual activity changes of the ventromedial prefrontal cortex during the judgments correlate with the degree of an individual's susceptibility to the framing information. CONCLUSION: We found (i) a neural correlate of a framing effect as postulated by behavioral decision theorists that (ii) reflects interindividual differences in the degree of the susceptibility to framing information. PMID- 16216689 TI - A behavioral window on the mind of the market: an application of the response time paradigm. AB - This article focuses on the role of implicit knowledge consumers have about particular brands, products or names. The major findings of several studies, conducted at the Mind of the Market Laboratory at Harvard Business School, are presented with specific emphasis on studies in which response time measurements were the core method. The results revealed that implicit measures provide a rich understanding of the meaning conveyed by a product or brand. Moreover, there is also considerable evidence that implicit measures may be better than traditional explicit measures as predictors of consumer behavior. We discuss the implications for the study of consumer behavior and the importance of combining several methods including neuroimaging, which has received recent attention by marketers, economists and social scientists. PMID- 16216690 TI - Arousal and consumer in-store behavior. AB - From a psychophysiological point of view, arousal is a fundamental feature of behavior. As reported in different empirical studies based on insights from theories of consumer behavior, store atmosphere should evoke phasic arousal reactions to attract consumers. Most of these empirical investigations used verbal scales to measure consumers' perceived phasic arousal at the point-of-sale (POS). However, the validity of verbal arousal measurement is questioned; self reporting methods only allow a time-lagged measurement. Furthermore, the selection of inappropriate items to represent perceived arousal is criticized, and verbal reports require some form of cognitive evaluation of perceived arousal by the individual, who might (in a non-measurement condition) not even be aware of the arousal. By contrast, phasic electrodermal reaction (EDR) has proven to be the most appropriate and valid indicator for measuring arousal [W. Boucsein, Physiologische Grundlagen und Messmethoden der dermalen Aktivitat. In: F. Rosler (Ed.), Enzyklopadie der Psychologie, Bereich Psychophysiologie, Band 1: Grundlagen and Methoden der Psychophysiologie, Kapitel, Vol. 7, Hogrefe, Gottingen, 2001, pp. 551-623] that could be relevant to behavior. EDR can be recorded simultaneously to the perception of stimuli. Furthermore, telemetric online device can be used, which enables physiological arousal measurement while participants can move freely through the store and perform the assigned task in the experiments. The present paper delivers insights on arousal theory and results from empirical studies using EDR to measure arousal at the POS. PMID- 16216691 TI - Reasoning or reacting to others? How consumers use the rationality of other consumers. AB - Consumers adapt their behavior to the structure of the information available in the environment where they form expectations. One factor in people's environments is other people. "Others" are a significant source of information and means of orientating oneself. In conditions such as uncertainty, insecurity, anxiety but also euphoria, it is "the others" who provide market participants with coordinates. The purpose of this paper is to pass forward an approach that relates to consumers' other-directedness and to demonstrate in what ways consumers use the rationality of other consumers. PMID- 16216692 TI - Discrepancy between effects of milligram and nanogram doses of a COX-2 inhibitor (celecoxib) on morphine state-dependent memory of passive avoidance in mice. AB - This experiment examined and compared the effects of pre-test administration of a selective COX-2 inhibitor (celecoxib), at the doses in the range of mg/kg and ng/kg on morphine state-dependent learning in step-down passive avoidance task in mice. Pre-training administration of 5mg/kg of morphine-impaired memory retrieval tested 24h later, which was restored by pre-test administration of the same dose of the drug. Pre-test administration of celecoxib (12.5, 25 and 50mg/kg), alone or in combination with morphine (1mg/kg) prevents morphine-induced memory impairment. Ultra-low doses (ULDs) of celecoxib (2, 10 and 50 ng/kg) produced no change in morphine-induced memory impairment. However, co-administration of nanogram doses of celecoxib with 5mg/kg of morphine in the test day prevented morphine-induced memory improvement, an action different from mg/kg doses. These findings implicate the involvement of COX-2 in memory retrieval and demonstrate that the effect of celecoxib ULD is different from that of mg/kg doses. PMID- 16216693 TI - Sensorimotor transduction of time information is preserved in subjects with cerebellar damage. AB - The cerebellar contribution to motor entrainment through rhythmic auditory stimuli was analyzed by comparing rhythmic motor responses in subjects with cerebellar pathologies and in healthy controls. Eleven patients with cerebellar lesions and eight healthy subjects tapped in synchrony with an auditory rhythmic stimulus using a hand-held pencil-shaped electrode connected to a PC. A 60 stimulus sequence was delivered with an ISI of 500 ms and changed at random to a new ISI value with either consciously perceived (+/-50 ms) or unperceived tempo changes (+/-10 ms). Synchronization patterns for both groups were computed based on the timing of inter-response intervals (IRIs) and synchronization errors (SE). Variability of IRI as well as the timing of adaptation patterns after the tempo changes were modeled and analyzed mathematically using a logistic/sigmoid function. Healthy subjects performed with significantly lower IRI variability than cerebellar patients. Patients with focal lesions performed with significantly lower IRI variability than patients with atrophic lesions. Asymptote parameters during isochronous synchronization as well as slope angles and symmetry points of the adaptation curves after tempo perturbation showed no significant differences between groups. Present data indicate that temporal variability of rhythmic motor responses is differentially affected by distinct cerebellar pathologies but that motor entrainment to auditory rhythms is not affected by lesion of the cerebellar circuits. PMID- 16216694 TI - Reversal of ethanol toxicity in embryonic neurons with growth factors and estrogen. AB - Prenatal exposure to ethanol is the cause of fetal alcohol syndrome, which is characterized by brain abnormalities and decreased mental capacity. In the current study, cultured neurons from embryonic rat cortices were used to study the reversal of ethanol toxicity on neuronal survival and neurite outgrowth. Ethanol treatment followed by treatment with estrogen and certain growth factors were used to assess the potential of these growth factors and estrogen to reverse the effects of ethanol damage. Cortical neurons from embryonic day (E) 16 rats were grown in defined medium with a glial plane at a distance of 1mm from the neurons. Ethanol (45 mM) was administered on day in vitro 1 (DIV 1) and DIV 4. Insulin-like growth factor-I (IGF-I, 10 ng/ml), insulin-like growth factor-II (IGF-II, 10 ng/ml), basic fibroblast growth factor (bFGF, 5 ng/ml), nerve growth factor (NGF, 100 ng/ml), and estrogen (Es, 10 ng/ml) were administered on DIV 4 and DIV 5. Cell viability was determined on DIV 6 using the intravital dyes fluorescein diacetate and propidium iodide. IGF-I and bFGF reduced ethanol's toxic effect on neuronal survival. Estrogen, bFGF, and NGF increased total neurite length after ethanol treatment. Although none of the treatments had a statistically significant effect on the mean number of primary neurites, all caused a statistically significant increase in the mean number of secondary neurites per cell (a measure of neuritic branching) relative to the ethanol treatment alone. PMID- 16216695 TI - Metallothionein provides zinc-mediated protective effects against methamphetamine toxicity in SK-N-SH cells. AB - Methamphetamine (METH) is a drug of abuse and neurotoxin that induces Parkinson's like pathology after chronic usage by targeting dopaminergic neurons. Elucidation of the intracellular mechanisms that underlie METH-induced dopaminergic neuron toxicity may help in understanding the mechanism by which neurons die in Parkinson's disease. In the present study, we examined the role of reactive oxygen species (ROS) in the METH-induced death of human dopaminergic SK-N-SH cells and further assessed the neuroprotective effects of zinc and metallothionein (MT) against METH-induced toxicity in culture. METH significantly increased the production of reactive oxygen species, decreased intracellular ATP levels and reduced the cell viability. Pre-treatment with zinc markedly prevented the loss of cell viability caused by METH treatment. Zinc pre-treatment mainly increased the expression of metallothionein and prevented the generation of reactive oxygen species and ATP depletion caused by METH. Chelation of zinc by CaEDTA caused a significant decrease in MT expression and loss of protective effects of MT against METH toxicity. These results suggest that zinc-induced MT expression protects dopaminergic neurons via preventing the accumulation of toxic reactive oxygen species and halting the decrease in ATP levels. Furthermore, MT may prevent the loss of mitochondrial functions caused by neurotoxins. In conclusion, our study suggests that MT, a potent scavenger of free radicals is neuroprotective against dopaminergic toxicity in conditions such as drug of abuse and in Parkinson's disease. PMID- 16216696 TI - TRPV1 desensitisation and endogenous vanilloid involvement in the enhanced analgesia induced by capsaicin in inflamed tissues. AB - The intra-plantar acute administration of 10 microg of capsaicin to mice which had received complete Freund's adjuvant (CFA) 1 week before inhibits the thermal inflammatory hyperalgesia it induces and even produces a long-lasting analgesia for at least 2 weeks. In this study, we show that the administration of capsaicin (10 microg) also reduces the immediate licking behavior evoked by the intra plantar administration of a lower dose of capsaicin (0.1 microg), the duration of this inhibitory effect being greater in CFA-inflamed mice (at least 2 weeks) than in non-inflamed animals (less than 4 days). Since this reduction of capsaicin induced licking behavior may be interpreted as a consequence of the transient receptor potential vanilloid 1 receptor (TRPV1) unresponsiveness, we conclude that the administration of 10 microg of capsaicin into inflamed tissues can render the TRPV1 desensitised. We next explored whether endogenous vanilloids released during inflammation contribute to maintain the analgesia triggered by exogenous capsaicin. The acute administration of capsazepine (10 microg; intra plantarly (i.pl.)) abolished the analgesic effect induced by the injection of capsaicin 1 week before in inflamed mice. From these results, it may be proposed that the maintenance by endovanilloids of the TRPV1 desensitisation induced by capsaicin could contribute to prolonging the analgesic effect induced by this agonist in inflamed tissues. PMID- 16216697 TI - Neuroprotective potential of dietary restriction against kainate-induced excitotoxicity in adult male Wistar rats. AB - The influence that dietary factors have on the nervous system and its susceptibility to disease, is an active area of biomedical research. Recent studies have shown that dietary restriction (DR) can have profound effect on brain function and vulnerability to injury and disease and can also enhance synaptic plasticity, which may increase the ability of brain to resist aging and restore function following injury. The dietary restriction may result in neuroprotection as suggested by marked reduction in neuronal cell death of the CA3 region of hippocampus after kainate administration in our study. We examined the effects of 3 months of DR (alternate day feeding regimen) on the antioxidants and antioxidant enzymes from different brain regions such as cerebral hemispheres, diencephalon, cerebellum and brain stem after kainate-induced excitotoxicity in adult male Wistar rats. The present study reports the beneficial effects of dietary restriction on different antioxidants and antioxidant enzymes against kainate-induced excitotoxicity in different brain regions of young adult male Wistar rats. The expression of stress response protein heat shock protein 70 (HSP 70) was also studied from discrete regions of rat brain under the same set of experimental conditions. DR significantly enhanced the expression of HSP 70 in kainic acid (KA)-treated rats, whereas KA treatment of ad libitum fed rats resulted in decreased HSP 70 expression. The DR was observed to exert neuroprotection by enhancing the expression of HSP 70 in kainic acid treated rats. PMID- 16216698 TI - Glycine modulates the center response of ON type rod-dominant bipolar cells in carp retina. AB - Effects of glycine on ON type rod-dominant bipolar cells (RBCs) were studied in isolated, superfused carp retina by intracellular recording technique and in carp retinal slice preparation by whole cell recording. Glycine of 4mM hyperpolarized RBCs and potentiated their light responses to large light spots, which was reversed by co-application of 10 microM strychnine. It was further found that illumination of the receptive field surround did not affect the depolarizing center response of RBCs. The above result therefore suggests that glycine modulates the center response of RBCs. Focal application of glycine to either dendrites or axon terminals of RBCs failed to induce any currents in both isolated cell and retinal slice preparations. On the other hand, glycine of 4mM increased the amplitude of the scotopic electroretinographic PIII component, which reflects the activity of rod photoreceptors. It seems likely that modulation by glycine of the RBC center response may be in part ascribed to a consequence of the potentiation of rod responses by glycine. PMID- 16216699 TI - Different effects of chronic exposure to ELF magnetic field on spontaneous and amphetamine-induced locomotor and stereotypic activities in rats. AB - The effects of chronic (7 days) exposure to an extremely low frequency magnetic field (ELF-MF, 50 Hz, 0.5 mT) on spontaneous and amphetamine-induced (1.5mg/kg, i.p.) locomotor and stereotypic activities in adult rats were examined by open field test for 2h on exposure days 1, 3, and 7. After 1 day of exposure to ELF MF, the spontaneous locomotor activity was increased clearly at the first hour of observation and significantly at the second one as compared to the corresponding values in other series with ELF-MF and sham-exposed animals. After 7 days of exposure to ELF-MF, an amphetamine enhancing effect on the locomotor activity was significantly reduced at the second hour of observation as compared to that in 1 day- and sham-exposed rats treated with amphetamine. In contrast to the locomotor activity, the amphetamine-induced stereotypic behaviour in 7-day pre-exposed rats was significantly reduced at the first hour versus sham-exposed rats. While at the second hour of observation this effect was significant as compared to 1- and 3-day exposed animals (but not sham-exposed rats). Our results indicate that an extremely low frequency magnetic field is able to affect differently two types of behaviour, which are dependent on both the time course of exposure and the imbalance in the brain mediatory systems. PMID- 16216700 TI - Interaction between M1-muscarinic and glutamatergic NMDA receptors on an inhibitory avoidance task. AB - It has been demonstrated that MK-801 potentiates the effects of the non-selective muscarinic antagonist scopolamine on memory in rats. In this study, we investigated the role of the M1-muscarinic receptor in this interaction, by administering different doses of dicyclomine (DIC) and MK-801 in combination to male Wistar rats before training on the inhibitory avoidance task. MK-801 and DIC in sub-effective doses were administered in combination. It was observed that MK 801 at a dose of 0.1125 mg/kg with a sub-effective dose of 8 mg/kg of DIC significantly impaired the retention test when compared with saline-treated animals, i.e. MK-801 potentiated the effects of dicyclomine on memory impairment. Our results suggest an important role for the M1-muscarinic receptor in the synergistic interaction between cholinergic muscarinic and glutamatergic NMDA receptors, which is in line with the findings that the interactive modulation between these two neurotransmitters systems constitutes an important mechanism in cognitive functions. PMID- 16216701 TI - Heat shock proteins protect both MPP(+) and paraquat neurotoxicity. AB - The exposure of immortalized rat neuroblast cells to MPP(+) and paraquat results in cell death. Heat shock pre-treatment prior to the addition of MPP(+) and paraquat significantly reduced cell death and led to an increase in the synthesis of Hsp 27 and Hsp70 proteins. Quercetin inhibits the synthesis of heat shock proteins (Hsp) and prevents their protective effect, which suggests that this protection was dependent on the Hsps synthesis. These data indicate that heat shock protects cells from the toxic effect of MPP(+) and paraquat. These results and the structural similarity between paraquat and MPP(+) support the role of paraquat as a putative risk factor in the etiology of Parkinson's disease. PMID- 16216702 TI - Illicit drug use, alcohol use and problem drinking among infrequent and frequent road ragers. AB - The purpose of this paper is to investigate the relationships between illicit drug and alcohol use, problem drinking, and road rage. Particular attention is devoted to the association between these behaviors and frequent involvement in road rage activities. The data are taken from the 2002 Centre for Addiction and Mental Health (CAMH) Monitor, a representative telephone survey with a sample of 2421 adults aged 18 and older in Ontario. A cluster analysis was performed and analysis of variance procedures were used to test for group differences. The cluster analysis revealed five distinct groups involved in various types of road rage behavior. Frequent road ragers, accounting for 5.3% of the sample, were involved in the most severe forms of road rage behavior and were most likely (24%) to report problem drinking and past year cannabis (23.8%), cocaine (5.4%), and ecstasy (10%) use. These data indicate that illicit drug use and alcohol problems are significantly greater for those involved in the most serious forms of road rage behavior. Further work is needed to identify the mechanisms by which illicit drug use and problem drinking are linked to road rage. PMID- 16216703 TI - Twelve-month prevalence and changes in driving after drinking: United States, 1991-1992 and 2001-2002. AB - BACKGROUND: Drinking and driving has been identified as one of the most important contributors of motor vehicle fatalities. This paper addressed the existing gap in our public health knowledge regarding the current prevalence of driving after drinking and how this has changed over the past decade. METHODS: Prevalence rates of drinking and driving in 2001-2002, and changes in those prevalence rates between 1991-1992 and 2001-2002 were examined in two large nationally representative surveys of the U.S. population. RESULTS: Overall, the prevalence of driving after drinking was 2.9% in 2001-2002 representing approximately six million U.S. adults. This rate was about three quarters of the rate observed in 1991-1992 (3.7%), reflecting a 22% reduction. Generally, the male-female differentials in the rate of driving after drinking decreased over the past decade. However, the sex ratios increased substantially for under-aged youth over the past decade, reflecting the sharp decrease in prevalence of driving after drinking among 18-20-year-old women. Constant and emerging subgroups at high risk for drinking and driving included Whites, Native Americans, males, under-aged young adults and 21-25-year-olds. CONCLUSIONS: The results of this study highlighted the need to continue to monitor prevalence and changes in driving after drinking. Results are discussed in the context of strengthening existing prevention and intervention efforts and developing new programs with the sociodemographic differentials observed in this study in mind. PMID- 16216704 TI - Postmortem diagnosis of hypertonic dehydration. AB - Beside morphological signs of hypertonic dehydration as tinting of skin, sunken eyes, dry surface of the galea or dry cutting areas of organs, a chemical profile of vitreous humor was proposed as a diagnostic tool for the diagnosis of hypertonic dehydration. The profile consists of an elevation of sodium >155 mmol/l, chloride >135 mmol/l and urea >40 mg/dl. This profile was named dehydration pattern. The value of this dehydration pattern for the diagnosis of hypertonic dehydration will be discussed by a short review of the literature and case reports. So far, the published literature on the dehydration pattern is not a sound scientific basis for the diagnosis of dehydration. PMID- 16216705 TI - Death due to neurogenic shock following gastric rupture in an anorexia nervosa patient. AB - We report a case of fatal gastric rupture discovered after death, which developed due to a bulimic attack of a 19-year-old woman suffering from anorexia nervosa. An autopsy revealed an acute gastric dilatation and rupture without commonly observed ischemic damage of gastric wall structures. However, it may be difficult to determine the cause of death despite the marked findings. The death as a consequence of neurogenic shock accounts for all the results of gross examination and histologic analysis. This case is the first reported case of fatal gastric rupture of an anorectic patient discovered after death. PMID- 16216706 TI - Sudden cardiac death in a 5-year-old girl associated with parvovirus B19 infection. AB - We report on a 5-year-old girl who suddenly collapsed and died while dancing at a family party. Histological examination of the heart including the cardiac conduction system revealed lymphocytic infiltrations of the sinu-atrial node and perivascular infiltration in the atrio-ventricular region. Additionally, foci of mononuclear infiltrates were observed in the myocardium. Consequently, myocarditis was diagnosed as cause of death. The child also had lymphocytic conjunctivis, parotitis and tracheitis. Evaluation of infections by means of nested polymerase chain reaction revealed parvovirus B19 DNA (PVB19) in tissue samples of the trachea. PMID- 16216707 TI - Evaluation of postmortem serum calcium and magnesium levels in relation to the causes of death in forensic autopsy. AB - There appears to be very poor investigation of postmortem serum calcium (Ca) and magnesium (Mg) for diagnostic evidence to determine the cause of death. The aim of the present study was a comprehensive analysis of the serum levels in relation to the causes of death in routine casework. Autopsy cases (total, n=360; 5-48 h postmortem), including blunt injury (n=76), sharp injury (n=29), asphyxiation (n=42), drownings (n=28: freshwater, n=11; saltwater, n=17), fire fatalities (n=79), methamphetamine (MA) poisoning (n=8), delayed death from traumas (n=37), and acute myocardial infarction/ischemia (AMI, n=61), were examined. In total cases, there was no significant postmortem time-dependent rise in serum Ca and Mg. Both Ca and Mg levels in the heart and peripheral blood were significantly higher in saltwater drowning compared with those of the other groups. In addition, a significant elevation in the Ca level was observed in freshwater drowning and fire fatalities, and in the Mg level in fatal MA intoxication and asphyxiation. Topographic analyses suggested a rise in serum Ca and Mg due to aspirated saltwater in drowning, that in serum Ca in freshwater drowning and fire fatalities of peripheral skeletal muscle origin and that in serum Mg in MA fatality and asphyxiation of myocardial and/or peripheral origin. These markers may be useful especially for diagnosis and differentiation of salt- and freshwater drownings and may be also helpful to determine the causes of death involving skeletal muscle damage, including burns and MA intoxication. PMID- 16216708 TI - Perianal and vulvar Crohn's disease presenting as suspected abuse. AB - Misdiagnosis of sexual abuse may arise in cases of vulvar and/or perianal diseases such as lichen sclerosus et atrophicus, Behcet's syndrome, bullous diseases, contact dermatitis, or neoplastic lesions. We present the case of a 7 year-old girl who was referred by her general paediatrician to the local hospital for perianal fissures and swelling of the labia majora. A report to the judicial authorities was made, for suspected sexual abuse, and the patient was transferred to the department of paediatric surgery. Here, histopathologic examination of vulvar and rectal biopsies revealed multiple non-caseating and non-confluent epithelioid-gigantocellular granulomas, consistent with a diagnosis of Crohn's disease, with cutaneous involvement of the genitalia. In cases of suspected sexual abuse, examination of children should be performed by a specialist in legal medicine in collaboration with a gynaecologist or paediatric surgeon. If the patient is hospitalised and the question of protection does not arise, physicians should exclude dermatological diseases before reporting to the judicial authorities. PMID- 16216709 TI - Accuracy of facial soft tissue thickness measurements in personal computer-based multiplanar reconstructed computed tomographic images. AB - The purpose of this study was to determine the precision and accuracy of facial soft tissue measurement using personal computer (PC)-based multiplanar reconstructed (MPR) computed tomography (CT) images and to evaluate the effect of the various CT scanning protocols on the facial soft tissue thickness measurement. Thirteen different CT imaging protocols were used to image a cadaver head. MPR reformations and three-dimensional (3D) reconstructions viewed on a laptop PC were used to make measurements at six specific sites on each set of images. These measurements were compared to physical measurements at the same sites. Increasing the slice thickness resulted in decreased image quality. Within the same slice thickness, increasing the pitch ratio in the spiral mode, resulted in decreasing image quality. The image quality of conventional CT scanning was relatively poorer than that of the spiral CT scanning. However, the mean deviation from the physical measurement was within 0.43 mm in every instance. This mean deviation was quite small and clinically acceptable for measuring the soft tissue thickness of the facial area. PC-based MPR CT images of the face using routine scanning CT protocols can be used to accurately measure soft tissue thickness in the facial region. However, for more fine and accurate data collection, scanning protocols with slice thicknesses less than 5mm, and a spiral/helical mode pitch less than 2:1 are recommended. PMID- 16216710 TI - Pistol image retrieval by shape representation. AB - Databases have been used to record data in forensic science, such as fingerprints, shoeprints, and photos. In traditional databases, we use "text" as the "keyword" for retrieving data (text-keyword retrieval); however, in some applications, "text" is not proper to describe the searching target, and in this case, "image" plays an important role. In this paper, we use "image features" as the "keywords", and show its potential for building up a prototype of pistol image databases. In current firearm databases of forensic science, the retrieval method is still by text-keyword retrieval. For experienced forensic examiners, this kind of databases may satisfy their requirement. However, for people who have little gun knowledge, how can they find the possible gun candidates or similar guns in the databases, if there are not any available words or marks on the gun? In this paper, we try to retrieve similar pistol images by the pistol shape instead of "text-keyword". This method can narrow down the searching range while identifying pistols by firearm databases. There are more than 300 pistol images in our pistol image database. From the experimental results, we can retrieve the similar pistol images in top five candidates for each pistol image. PMID- 16216711 TI - Legal perceptions of forensic DNA profiling part I: a review of the legal literature. AB - A forensic biologist is usually involved in the criminal justice system process somewhere between the police and the legal system, interacting in a practical context regularly and extensively with both. Forensic DNA research and development commonly involves initiatives that encroach into the neighbouring domains of the law enforcement or legal agencies. Despite this level of association, establishing meaningful cross-disciplinary communication and understanding within the justice system remains a challenge. As an example, there is an abundance of literature relating to forensic DNA profiling in legal and criminological periodicals. Such journals are perhaps outside the regular reading of forensic scientists and much of the legal discussion appears to go unnoticed. This situation is understandable; however, it is also undesirable particularly as forensic DNA developments are intertwined with significant changes in legislation and contentious issues of privacy, civil liberty and social justice. This paper attempts to address this shortcoming directly by summarising - from the viewpoint of a forensic scientist - some of the discussion in the legal literature. In particular the review focuses on discussion raising ideological and ethical concerns. Awareness of these views is of relevance to forensic science. It assists us to accurately place DNA evidence into context and to develop its role in achieving the broader criminal justice system objectives. Understanding the discussion also provides a way to enter the debate and communicate at an appropriate level the true potential of DNA to the legal community. PMID- 16216712 TI - Postfeeding radial dispersal in larvae of Chrysomya albiceps (Diptera: Calliphoridae): implications for forensic entomology. AB - Blowflies utilize discrete and ephemeral breeding sites for larval nutrition. After the exhaustion of food, larvae begin dispersing in search of sites to pupate or additional food sources, a process referred as postfeeding larval dispersal. Some of the most important aspects of this process were investigated in the blowfly Chrysomya albiceps, employing a circular arena to allow radial dispersion of larvae from the center. The results showed a positive correlation between burial depth and distance, and a negative correlation between distance and pupal weight. These results can be used in forensic entomology for the postmortem interval estimation of human corpses in medico-criminal investigations. PMID- 16216713 TI - A study of mutation rates and the characterisation of intermediate, null and duplicated alleles for 13 Y chromosome STRs. AB - Previously reported Y chromosome STR haplotype databases for three UK population groups, plus additionally analysed samples, have been scrutinised for the presence of non-standard (intermediate, null and duplicated) alleles. These alleles have been characterised by sequencing, some showing changes in the repeat structure, and the frequencies reported. Mutation rates for each of the 13 STRs have been calculated when analysis of father-son pairs has been possible. An example illustrating the use of non-standard alleles in a large family tree is outlined. PMID- 16216714 TI - Development of two pentaplex systems with X-chromosomal STR loci and their allele frequencies in a northeast German population. AB - In this study we present two new pentaplex systems for the coamplification of X chromosomal short tandem repeats (STRs). X-penta-1 comprises DXS9898, DXS6807, HPRTB, DXS101, and androgen receptor (ARA); X-penta-2 consists of DXS7133, DXS10011, DXS7424, DXS8377, and DXS8378. In addition, allele frequencies for these loci in a northeast German population comprising 100 females and 105 males were shown. The applicability and usefulness of our two PCR pentaplex approaches in paternity deficiency cases is demonstrated by a combined power of discrimination (PD(c)) for both females and males with PD(c)>0.999999. PMID- 16216715 TI - Population data on 15 autosomal STRs in a sample from East Timor. AB - Allele frequencies for the fifteen STRs included in the AmpF/STR Identifiler (CSF1PO, D2S1338, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D19S433, D21S11, FGA, TH01, TPO and VWA) were estimated from a sample of 186 unrelated individuals from East Timor. No deviations from Hardy-Weinberg equilibrium were observed (only after applying the Bonferroni correction in the cases of D2S1338, TPO and D5S818). Genetic parameters of forensic interest were calculated and comparison with geographically nearby populations was performed. PMID- 16216716 TI - Validation of a first-generation long-oligonucleotide microarray for transcriptional profiling in the pig. AB - A first-generation porcine oligonucleotide set, representing 13,297 cDNAs and ESTs, has been designed by Qiagen-Operon for transcriptional profiling. To validate this set, microarrays containing each 70-mer oligonucleotide, referred to as the Qiagen-NRSP8 array, were hybridized with targets from porcine adult liver, lung, muscle, or small intestine. Transcriptome analyses showed that 11,328 of the oligonucleotides demonstrated expression in at least one tissue. Statistical analyses revealed that 1810 genes showed differential expression among tissues (Bonferroni adjusted p < 0.05). Biological pathways identified by DAVID/EASE analysis using a list of 423 tissue-selective genes matched archetypal pathways in the corresponding human or mouse tissue. Real-time quantitative PCR confirmed expression patterns for 9 of 11 genes tested. Our results demonstrate that this first-generation porcine oligonucleotide array is informative and the specificity is high. This is essential validation for investigators using the Qiagen-NRSP8 array for porcine functional genomics and for using the pig in modeling important physiological problems. PMID- 16216717 TI - Lipoprotein(a): Looking ahead. PMID- 16216718 TI - Improving guidelines for cardiovascular practice? PMID- 16216719 TI - Isocaloric substitution of plant sterol-enriched fat spread for carbohydrate-rich foods in a low-fat, fibre-rich diet decreases plasma low-density lipoprotein cholesterol and increases high-density lipoprotein concentrations. AB - BACKGROUND AND AIM: The aim of the study was to determine the effects on plasma cholesterol of replacing a plant sterol-enriched fat spread with carbohydrate rich foods relative to a diet high in saturated fat. METHODS AND RESULTS: Twenty nine men and women, from the general community, with mean age (SD) 48 (14)y, body mass index 29.0 (6.2)kg/m(2), and plasma total cholesterol concentration 6.48 (0.97)mmol/L completed the randomised, crossover dietary intervention. There were three diets: New Zealand diet (NZ diet) high in total (34%kJ) and saturated (15%kJ) fat, a cholesterol-lowering fibre-rich diet reduced in total (30%kJ) and saturated fat (8%kJ) but including a plant sterol spread (PS diet), and the same cholesterol-lowering diet with the plant sterol spread isocalorically replaced with carbohydrate (CHO diet); total fat, 26%kJ; saturated fat 7%kJ. All foods were provided and each diet was followed for four weeks. Mean (SD) plasma low density lipoprotein cholesterol concentration declined from 4.68 (0.91)mmol/L on the high saturated fat diet to 4.12 (0.83)mmol/L (P<0.001) on the carbohydrate diet and 3.76 (0.84)mmol/L (P<0.001) on the plant sterol diet. The 20% decrease on the plant sterol diet was significantly greater (P<0.001) than the 12% decrease on the carbohydrate diet. Relative to the NZ diet, mean (95% CI) plasma high-density lipoprotein cholesterol concentration changed by -0.11 (-0.16, 0.06)mmol/L on the CHO diet but was not different at the end of the PS diet, 0.03 (-0.09, 0.02). CONCLUSION: Including a plant sterol-enriched fat spread in a cholesterol-lowering diet produces a more favourable plasma lipid profile than the same diet made lower in total and saturated fat by replacing the spread with carbohydrate-rich foods. PMID- 16216720 TI - Alpha-linolenic acid and heart rate variability in women examined for coronary artery disease. AB - BACKGROUND AND AIM: Some studies suggest that alpha-linolenic acid (ALA) may protect against coronary artery disease (CAD) and CAD related death (i.e. sudden cardiac death). Thus, some limited data suggest an antiarrhythmic effect of ALA. We therefore investigated the association between ALA and 24-h heart rate variability (HRV), a strong predictor of arrhythmic events and sudden cardiac death, in women. METHODS AND RESULTS: The content of ALA in adipose tissue and HRV was determined in 106 women referred for coronary angiography due to suspected CAD. Mean age was 59.5 years (+/-7) and the content of ALA in adipose tissue was 0.9% (+/-0.2). Smokers (n=33) had significantly lower HRV compared to non-smokers, whereas the two groups were otherwise comparable regarding clinical characteristics. If the patients were divided according to SDNN quartiles (the major HRV parameter), women belonging to the lowest quartile had the lowest ALA level (0.8% vs 0.9%, p < 0.01). Furthermore, a positive correlation was found between ALA and HRV, especially among smokers (r=0.5, p < 0.001). Linear multiple regression analysis revealed that ALA was independently and positively associated with HRV. CONCLUSIONS: A positive and independent association was found between ALA in adipose tissue and HRV in women referred for coronary angiography due to suspected coronary artery disease. This association was even stronger for women who smoked. The results might suggest a possible antiarrhythmic effect of ALA in women, an effect which could explain results from previous studies. However, intervention trials with ALA in humans, not least women are warranted. PMID- 16216721 TI - Consumption of argan oil may have an antiatherogenic effect by improving paraoxonase activities and antioxidant status: Intervention study in healthy men. AB - BACKGROUND AND AIM: Due to its high antioxidant and mono- and polyunsaturated fatty acid content virgin argan oil (VAO) could play a beneficial role in cardiovascular prevention. We were therefore interested in determining whether the consumption of VAO could improve plasma paraoxonase (PON1) activities and antioxidant status in healthy men. METHODS AND RESULTS: Sixty young men were included in this interventional study. They were given a controlled diet for 2 weeks as baseline and then received 25 g/day of butter. The group was randomised to two diet group periods of 3 weeks each. The VAO group received 25 ml/day of oil and the extra virgin olive oil (EVO) group received the same quantity of EVO as control group. Plasma PON1 activities, antioxidant vitamins and LDL susceptibility to oxidation were measured. The analysis of the results shows that PON1 activities increase significantly in both groups and that lipoperoxides and conjugated dienes formation decreases significantly in VAO and EVO groups compared to baseline values (P=0.001 and P=0.014, respectively). Vitamin E concentration increases significantly only in VAO group (P=0.007). Susceptibility of LDL to lipid peroxidation shows a significant increase in lag phase and a significant decrease in maximum diene production in VAO (P=0.005) and EVO groups (P=0.041 and P=0.005, respectively). CONCLUSIONS: Our findings confirm the beneficial effect of EVO on plasma antioxidant status and show for the first time the same effect for VAO supplementation in man. Thus, VAO offers an additional natural food supplement to reduce cardiovascular risk. PMID- 16216722 TI - Family history, diabetes and extension of coronary atherosclerosis are strong predictors of adverse events after PTCA: A one-year follow-up study. AB - BACKGROUND AND AIM: In this study we addressed some open questions in patients with coronary artery disease (CAD). First, we analysed which of the traditional risk factors was associated with the spreading of coronary stenosis and second, we aimed to identify if any variable was predictive of post-percutaneous transluminal coronary angioplasty (PTCA) clinical events. METHODS AND RESULTS: We collected a consecutive series of patients with CAD (n=301) and in the subgroup of patients undergoing PTCA (n=135) we performed a prospective one-year follow-up study recording cardiovascular morbidity and total mortality. According to the extension of coronary atherosclerosis, we found a significant relationship with the prevalence of diabetes in men and with plasma HDL-cholesterol concentrations in women. The follow-up was completed in 95% of patients; we did not document any death whereas clinical events were registered in 16% of patients. At univariate analysis, we found that patients with clinical events had a higher prevalence of family history of CAD (43% vs 14%, p<0.005), diabetes (52% vs 21%, p<0.005) and multivessel disease (52% vs 35%, p<0.05). Multivariate analysis (logistic regression) confirmed that family history of CAD (OR 4.6, 95% CI 1.7-12.8, p<0.005), diabetes (OR 4.0, 95% CI 1.5-10.6, p<0.01) and multivessel disease (OR 2.8, 95% CI 1.1-7.4, p<0.05) were the only variables predictive of clinical events. CONCLUSIONS: In this study, factors associated with the spreading of coronary stenosis were different according to the gender. Moreover, the presence of diabetes and multivessel disease had a negative impact on the long-term prognosis of patients undergoing PTCA. In addition, the family history of CAD represented in our study a strong predictor of clinical events. We suggest that in the management of post-PTCA patients, the role of individual baseline clinical characteristics must be taken into account and that subjects with a family history of premature CAD, diabetes and a wide extension of coronary disease represent those with the highest risk. PMID- 16216723 TI - Association of LDL cholesterol with carotid atherosclerosis in menopausal women affected by the metabolic syndrome. AB - BACKGROUND AND AIM: The metabolic syndrome is a highly prevalent condition associated with cardiovascular disease. However, the contribution of LDL to cardiovascular risk is not estimated since it is not part of ATP III criteria. METHODS AND RESULTS: This is an observational study evaluating the association between metabolic syndrome and carotid atherosclerosis, according to LDL cholesterol levels. Two hundred and sixty-five menopausal women were consecutively enrolled, they all underwent clinical examination, biochemical characterization and ultrasound evaluation. In particular, carotid atherosclerosis, a well known marker of cardiovascular disease, was evaluated. Women affected by cardiovascular disease were excluded from the study. The metabolic syndrome was found strongly associated with carotid atherosclerosis in our study population. In individuals with normal or near normal LDL, the incidence of carotid atherosclerosis was significantly lower than in subject with high LDL. A high plasma LDL concentration was independently associated with carotid atherosclerosis (p=0.026) among women with the metabolic syndrome. CONCLUSIONS: High LDL cholesterol levels are associated with carotid atherosclerosis in menopausal women with the metabolic syndrome. Although it remains prudent to recommend an integrated control of all modifiable risk factors to prevent cardiovascular disease, decreasing LDL levels should be considered a high priority. PMID- 16216724 TI - Implications of emerging risk factors for therapeutic intervention. AB - Recently, the National Cholesterol Education Panel (NCEP) of the United States of America commented on the implications of new clinical trials for the Adult Treatment Panel III (ATP III) guidelines. In this commentary, new categories of "moderately high" and "very high" coronary risk were proposed with new "therapeutic options" for low-density lipoprotein (LDL) cholesterol of < or = 100 mg/dL and < or = 70 mg/dL respectively. In ATP III, these "moderately high" risk patients had been classified as moderate risk with an LDL treatment goal of < or = 130 mg/dL, while the "very high" risk patients had been classified as high risk with a treatment goal of < or = 100 mg/dL. Risk classification in the new NCEP publication is based essentially on the combination of the Framingham risk score plus counting of classical risk factors. In the present document, the International Task Force for Prevention of Coronary Heart Disease responds to this NCEP commentary and supports the suggestion of more intensive LDL cholesterol lowering in particular cases. However, the Task Force feels that a classification based on a combination of a risk score plus a count of emerging risk factors is a more logical way to identify such patients requiring lower LDL cholesterol levels than a scheme in which classical risk factors are taken into account twice, once in a count and once in a risk score. PMID- 16216725 TI - Arterial hypertension and cardiovascular prognosis after successful repair of aortic coarctation: a clinical model for the study of vascular function. AB - Despite successful surgical repair, aortic coarctation is associated with unfavourable prognosis mainly due to cardiovascular disease. Late timing of repair and arterial hypertension represent adverse prognostic factors. Arterial hypertension can recur after coarctation repair, despite the absence of residual obstruction, with a prevalence of up to 45%. Furthermore, even subjects with normal blood pressure values at rest may show an abnormal blood pressure elevation during exercise and daily life activities. The pathophysiology of such abnormal blood pressure behaviour is unclear. Different mechanisms have been proposed: resetting of the renin-angiotensin system, neurological dysfunction and impaired vascular reactivity and/or elastic properties. Several studies have supported these hypotheses, although the suggestion of a causative role of vascular dysfunction persisting late after coarctation repair has recently become more popular. Further studies are needed to investigate this issue; this particular syndrome may represent an important study model for the understanding of systolic hypertension. PMID- 16216726 TI - Effect of oxygen, ozone and hydrogen peroxide bleaching stages on the contents and composition of extractives of Eucalyptus globulus kraft pulps. AB - The effects of oxygen (O), ozone (Z) and hydrogen peroxide (P) bleaching stages on the composition and total amount of Eucalyptus globulus kraft pulp lipophilic extractives was studied. These bleaching stages led to the partial removal and to several oxidative transformations of fatty acids and sterols, the main lipophilic extractives found in the unbleached pulp. Unsaturated extractives were found to be partially degraded while saturated ones were, in general, stable. The oxygen and hydrogen peroxide bleaching stages were more effective than ozone in removing fatty acids from pulp, by dissolution in the liquid phase. On the other hand, the ozone stage was more effective in the oxidative degradation of sterols. Oxygen and hydrogen peroxide bleaching stages were also effective in sterols removal, but led to the formation of sterol oxidation derivatives, previously shown to be involved in the formation of pitch that accumulates in the bleaching filtrates. PMID- 16216727 TI - Olive bagasse (Olea europea L.) pyrolysis. AB - Olive bagasse (Olea europea L.) was pyrolysed in a fixed-bed reactor. The effects of pyrolysis temperature, heating rate, particle size and sweep gas flow rates on the yields of the products were investigated. Pyrolysis runs were performed using pyrolysis temperatures between 350 and 550 degrees C with heating rates of 10 and 50 degrees C min(-1). The particle size and sweep gas flow rate varied in the ranges 0.224-1.8mm and 50-200 cm3 min(-1), respectively. The bio-oil obtained at 500 degrees C was analysed and at this temperature the liquid product yield was the maximum. The various characteristics of bio-oil obtained under these conditions were identified on the basis of standard test methods. The empirical formula of the bio-oil with heating value of 31.8 MJ kg(-1) was established as CH(1.65)O(0.25)N(0.03). The chemical characterization showed that the bio-oil obtained from olive bagasse may be potentially valuable as a fuel and chemical feedstock. PMID- 16216728 TI - Willow growth in response to nutrients and moisture on a clay landfill cap soil. I. Growth and biomass production. AB - The growth and biomass production by willow (Salix viminalis L.) was studied in lysimeters containing Oxford clay landfill cap soil with different amendments, bulk densities and watering regimes. Three years from planting, stem biomass in well-watered plants was least (0.28 kg plant(-1)) with high bulk density soil (1480 kg m(-3)) and no nutritional amendment but was increased 10-fold (2.53 kg plant(-1)) by reducing soil bulk density (1200 kg m3) and adding amendments. In comparison, on a sandy loam soil it was 6.23 kg plant(-1). There were similar differences in number of stems plant(-1), stem basal area plant(-1) and plant leaf area which can be attributed to low nitrogen and phosphorus levels in Oxford clay. Water stress reduced stem biomass production by 26-37% and caused higher root:stem ratios. These were also higher on Oxford clay than on the sandy loam. Successful biomass production from willow on Oxford clay landfill caps will therefore require nutritional amendment. PMID- 16216729 TI - Willow growth in response to nutrients and moisture on a clay landfill cap soil. II: Water use. AB - Water use by willow (Salix viminalis L.) was studied in lysimeters containing clay landfill cap and sandy loam soils under different watering and amendment regimes. With plentiful water and amendments, seasonal ET increased annually and was highest in the sandy loam, increasing from 360 l plant(-1) in the establishment year to almost 1200 l plant(-1) in the third year. Seasonal ET was highly correlated with leaf area duration. Amendment of Oxford clay resulted in increases in plant leaf area, dry matter production and seasonal ET. Water stress reduced seasonal ET by 10-14% in the second year and 25-41% in the third. Water use efficiency was low for the un-amended clay treatment (1.4 g kg(-1)) but was similar in the amended clay (5.0 g kg(-1)) and sandy loam (4.9 g kg(-1)). This highlights the interdependence of water use and biomass production in willow. PMID- 16216730 TI - Manganese peroxidase production in submerged cultures by free and immobilized mycelia of Nematoloma frowardii. AB - The agaric basidiomycete Nematoloma frowardii has been suggested as a good alternative for production of the extracellular ligninolytic enzyme, manganese dependent peroxidase (MnP). Some cultural and environmental factors influencing the enzymatic activity in shaken flasks and aerated fermenter cultures were evaluated to improve the yields of the process. A low nitrogen medium (1.36 mM N added as ammonium tartrate), containing 16 g/l glucose (C/N ratio=65.3), 2mM Mn2+ and inoculated with immobilized polyurethane foam mycelium, made it possible to obtain a MnP yield of 2304 nkat/l in 8 days. Under these operational conditions, the enzyme productivity in the immobilized cells of N. frowardii was 1.4 times higher than that obtained with the free fungus. In the procedure with the reusable immobilized mycelium (semi-continuous culture) as many as three subsequent 10 day batches could be fermented by using the same carrier with no loss of MnP activity. PMID- 16216731 TI - Fungal solid state culture of palm kernel cake. AB - Palm kernel cake (PKC), an agro-industrial by-product used extensively in the animal feed industry, has limited use in fish feeds due to its high fiber and low protein contents. In this study, PKC was processed under solid state culture conditions with five fungal strains and the effect of this fungal culturing on the amino acid, fatty acid, cellulose and hemicellulose fractions was evaluated. Fungal strains used were Sclerotium rolfsii, Trichoderma harzianum, Trichoderma longiobrachiatum, Trichoderma koninggi and Aspergillus niger. Fungal growth was carried out at 50% moisture level and 1% inoculum level for 7 days. A significant increase in protein content from 18.76% to 32.79% was obtained by growing T. longibrachiatum on PKC. Cellulose level decreased significantly from 28.31% to 12.11% for PKC cultured with T. longibrachiatum, and hemicellulose from 37.03% to 19.01% for PKC cultured with A. niger. Fungal culturing of PKC brought about an increase in the level of unsaturated- and a decrease in the level of the saturated-fatty acids. PMID- 16216732 TI - Arsenic removal from groundwater by pretreated waste tea fungal biomass. AB - Arsenic contamination in ground water poses a serious threat on human health. The tea fungus, a waste produced during black tea fermentation has been examined for its capacity to sequester the metal ions from ground water samples. Autoclaved tea fungal mat and autoclaving followed by FeCl3 pretreated tea fungal mat were exploited for removal of As(III), As(V) and Fe(II) from ground water sample collected from Kolkata, West Bengal, India. The biosorption rate tends to increase with the increase in contact time and adsorbent dosage. FeCl3 pretreated and autoclaved fungal mats removed 100% of As(III) and Fe(II) after 30 min contact time and 77% of As(V) after 90 min contact time. The optimum adsorbent dosage was 1.0 g/50 mL of water sample. The results revealed that the FeCl3 pretreated fungal mat could be used as an effective biosorbent for As(III) and As(V); autoclaved fungal mat for Fe(II) removal from ground water sample. PMID- 16216733 TI - Biosorption of dyes using dead macro fungi: effect of dye structure, ionic strength and pH. AB - Biosorbents prepared from dead macro fungi, namely Fomes fomentarius and Phellinus igniarius, were applied for the uptake of Methylene Blue (MB) and Rhodamine B (RB). Equilibrium isotherm data could be well described by the Langmuir and Freundlich models. Methylene Blue was found to be more adsorbable than Rhodamine B. Langmuir monolayer coverage was determined as 204.38-232.73 mg/g and 25.12-36.82 mg/g for MB and RB, respectively. Molecular structure and ionic radius of dyes were found to be responsible for differences in their uptakes. Results showed that sorption of MB increased while that of RB decreased as pH of respective dye solutions changed from 3 to 11. An increase in ionic strength also exhibited an adverse effect on dye sorption capacity. Ionic strength and pH affected the sorption of MB more as compared to the sorption of RB. The presence of carboxylic (-ve) and amino (+ve) groups in RB could explain the lower sorption of RB compared to MB. PMID- 16216734 TI - Characterization of a phosphorus-potassium solution obtained during a protein concentrate process from sunflower flour. Application on rye-grass. AB - The process that permits the ability to obtain a protein extract from defatted sunflower flour also produces a solution very rich in phosphorus (P) and potassium (K), which also contains small concentrations of humic substances. The aim of this study has been to determine the possible agricultural use of this extract. Therefore the phosphorus-potassium solution (experimental solution) was analyzed to determinate its pH and its content of nitrogen, proteins, organic carbon, humic substances potassium and phosphorous. The experimental solution was applied on rye-grass (Lolium multiflorum Lam.) and afterwards the results were analyzed we calculated the germination percentage and the fresh and dry weights that were obtained after each cut throughout the duration of the experiment. In addition the different pigment types (chlorophyll a, chlorophyll b and carotenoids) were quantified. The conclusions of the study examine how this time stable experimental solution improves the long-term effects and also the level of pigments, especially carotenoids, of the plants that have been treated. PMID- 16216736 TI - Can sonography be used to help differentiate between radial scars and breast cancers? AB - The aim of this study was to determine whether sonography can help differentiate radial scars (RS) from breast cancers. Series of 75 consecutive mammographic screen-detected RS and carcinomas were reviewed: shape, orientation, echotexture, halo, acoustic attenuation and architectural distortion were compared for sonographic RS and cancers. RS were 43% sonopositive (25/58 examined) vs. 93% sonopositive carcinomas (68/73 examined); P<0.001. Of 22 RS and 66 cancers available for film review, findings were: echogenic halo in zero RS vs. 38 cancers (0% vs. 58%); tiny sonographic cysts in 3 RS vs. zero carcinomas (14% vs. 0%); assessment category malignant vs. indeterminate/suspicious (8% vs. 59%, P<0.001); breast architecture disruption (43% vs. 91%, P<0.001); sound attenuation (55% vs. 86%, P<0.005), taller-than-wide shape (36% vs. 56%, P=0.11). RS showed echogenic components more than cancers (32% vs. 9%, P=0.016). Jagged margins were equally seen (77% vs. 76%, P=0.89). The findings suggest that sonographic differences can help discriminate between RS and carcinomas. PMID- 16216737 TI - The elevated level of CXCR4 is correlated with nodal metastasis of human breast cancer. AB - CXCR4, the receptor for stromal cell-derived factor-1(SDF-1), belongs to the chemokine receptor family and has been shown to play an important role in regulating the directional migration of breast cancer cells to sites of metastasis. In the present study, we evaluated the expression of CXCR4 and its association with pathological features and clinical outcome in human breast cancer. Expression of CXCR4 in eight breast cancer cell lines and breast cancer tissues was investigated using conventional PCR. Levels of CXCR4 transcript and protein were examined in human breast cancer tissues (n=120) and corresponding normal tissues (n=32) using real-time quantitative PCR and immunohistochemistry, respectively. The level of CXCR4 expression was analyzed against tumour types, grade, nodal status, recurrence, metastasis, and survival over a median 120 month follow-up period. The expression of CXCR4 was detected in all breast cancer cell lines examined, as well as in breast cancer tissues and breast normal tissues. Breast cancer tissues highly expressed CXCR4 compared with corresponding normal tissues (P=0.029). The level of CXCR4 expression showed a significant difference between node-positive group and node-negative group (19+/-13 vs. 49.7+/-9, respectively, P=0.03). The level of CXCR4 expression was marginal, yet statistically insignificant, higher in tumours from patients with metastatic disease compared with those who remained disease free. No correlation was seen between levels of CXCR4 and the overall survival, although at higher levels of CXCR4 linked to shorter disease free survival (113.0 vs. 136.7 months in patients with low CXCR4, P=0.14, Cox proportional test). The level of CXCR4 expression is significantly correlated with lymph node metastasis. The elevated levels of CXCR4 suggest that the patient has high possibility of lymph node metastasis. CXCR4 may be a useful prognostic indicator and a potential therapeutic target in cancer therapies in patients with breast cancer. PMID- 16216738 TI - Reduced dose capecitabine is an effective and well-tolerated treatment in patients with metastatic breast cancer. AB - Fifty-seven patients with metastatic breast cancer have been treated with reduced dose capecitabine 1g/m2 twice daily for 14 days repeated every 3 weeks after failure of a number of chemotherapy regimens or hormonal treatment. The overall objective response rate was 28% with 2% complete response rate and 26% partial response. Twenty nine percent objective response rate was achieved in 17 patients pretreated with anthracyclines and taxanes. The median duration of response and survival were 8.5 and 9 months, respectively. One and two year overall survival probabilities were 37% and 20%, respectively. The most common treatment-related adverse events (all grades) were hand-foot syndrome (HFS) (32%), nausea (21%) and diarrhoea (19%). Dose limiting toxicities, especially HFS were rare. Within the limits of phase II trial, this reduced dose of capecitabine appears as effective for advanced breast cancer as full dose capecitabine with a lower incidence of toxicity. PMID- 16216739 TI - Management of lactational breast abscesses. AB - The purpose of the present prospective study was to compare incision and drainage against needle aspiration for the treatment of breast abscesses in lactating women. During the 3-year study period, patients with breast abscesses were randomized 1:1 to undergo either incision and drainage (23 patients) or needle aspiration (22 patients). Ultrasound guidance was not used for any of these patients. Age, parity, localization of abscess, whether or not nipples were cracked, duration of symptoms and lactation, abscess diameter, pus culture results, breast infection history during any previous period of lactation, healing time, recurrence, cosmetic outcome in the case of incision and drainage, and volume of pus removed and number of aspirations needed in the case of aspiration were recorded. The treatment value of each of these techniques was investigated. Student's t-test, Fisher's exact test, a Chi-square test and the Mann-Whitney U-test were used for statistical analysis. In the incision and drainage group all patients were treated successfully, but 1 patient (4%) had a recurrence 2 months after complete healing and 16 patients (70%) in this group were not pleased with the cosmetic outcome. In the needle aspiration group, overall 3 patients were treated with a single aspiration and 10 patients (45%) with multiple aspirations, but 9 patients (41%) did not heal following needle aspiration and subsequently required incision and drainage in addition. No recurrences were observed in the needle aspiration group during the follow-up period. The risk factors for failure of needle aspiration for breast abscesses were abscesses larger than 5 cm in diameter, unusually large volume of aspirated pus, and delay in treatment. In conclusion, breast abscesses smaller than 5 cm in diameter on physical examination can be treated with repeated aspirations with good cosmetic results. Incision and drainage should be reserved for use in patients with larger abscesses. PMID- 16216740 TI - Response of bilateral choroidal metastases of breast cancer to therapy with trastuzumab. AB - Intraocular metastases, especially those of the choroidal plexus, are not common in metastatic breast cancer patients and are typically associated with a poor prognosis and impaired quality of life. A 45-year-old woman with breast cancer overexpressing HER2 and metastasizing to choroidal plexus, lymph nodes and skin received a combination of trastuzumab and paclitaxel as first-line treatment. Subsequently, at progression, trastuzumab was reintroduced together with vinorelbine. Administration of trastuzumab with either paclitaxel or vinorelbine led to a rapid improvement of the ocular symptoms, associated with a rapid objective response of all metastatic lesions and a prompt improvement in the quality of life. Choroidal metastases from breast cancer overexpressing HER2 are responsive to trastuzumab and chemotherapy (paclitaxel or vinorelbine). The susceptibility of ocular metastases to this approach seems different to that of other sanctuary disease sites. PMID- 16216741 TI - A case of nodular fascitis of the breast and review of the literature. AB - Nodular fascitis is a benign condition commonly affecting the limbs. In the breast, however, it may mimic Breast Cancer clinically and radiologically. We report a case of nodular fascitis of the breast and its conservative management. With the use of cytology, core biopsy and imaging we were able to confidently diagnose nodular fascitis and, therefore, avoid surgical intervention. Recent literature is reviewed demonstrating the benign nature of this disease. PMID- 16216742 TI - Extramammary Paget's disease of the breast: an unusual location with prognostic implications. AB - Extramammary Paget's disease is a neoplastic process of intraepidermal origin; its management and prognosis are very different from those of mammary Paget's disease. We present a case of extramammary Paget's disease located in the breast. This is an exceptional location, whose differential diagnosis is important because of its therapeutic and prognostic implications. PMID- 16216743 TI - Breast carcinoma in diabetic mastopathy. AB - Diabetic mastopathy, or diabetic fibrous breast disease is a well-characterised benign, fibro-inflammatory condition affecting women with insulin dependent diabetes. To date the relationship between this condition and breast carcinoma has been poorly reported. We describe a case of breast carcinoma arising within a diabetic fibrous breast lesion, in a renal transplant recipient. PMID- 16216744 TI - Tumour localisation with a metal coil before the administration of neo-adjuvant chemotherapy. AB - Complete clinical response (CR) with tumour disappearance is not uncommon after neo-adjuvant chemotherapy (NAC) for locally advanced breast cancer, avoiding 25% mastectomies by facilitating breast-conserving procedures. We reviewed our series to understand the feasibility and utility of marking the cancer site before administering NAC. In total, 23 women (median age 47 years) with T2-4, N0-1, M0 tumours were considered unsuitable for breast conserving surgery between January 2002 and November 2003, thus received NAC following a coil placement at the core of tumour. All patients had the coil successfully inserted and no migration or infection was recorded. Eight patients (35%) had a radiological CR (rCR) including 3 (13%) with pathological CR (pCR). In total, 87% patients were managed conservatively. The insertion of a metal coil is a simple mean to provide a landmark for localisation and excision when the breast lump becomes impalpable and radiologically undetectable after the administration of NAC. PMID- 16216745 TI - Management of breast cancer in patients prenatally exposed to diethylstilbestrol: are we prepared? AB - The use of diethylstilbestrol (DES) for high risk pregnancy has exposed millions of mothers to an increased risk of breast cancer, and also resulted in a generation of women with genital tract abnormalities, such as vaginal adenosis. It is still too early to say that exposure to DES will also result in an increased risk of breast cancer in the offspring, though there is some preliminary evidence to support this. The employment of optimal hormonal therapy (for breast cancer) in this special population may be hampered by the fact that agents with oestrogen agonistic activity (such as tamoxifen) may be contraindicated. Though some of the newer hormonal agents, such as the pure anti oestrogen Fulvestrant and the aromatase inhibitors, could be considered interesting alternatives for postmenopausal patients, their safety in this population has never been evaluated. Finally, the prevalence prenatal exposure to DES may have been underestimated patients diagnosed with breast cancer, though this information might have major implications in their management. We report on the interesting example of a young woman with a history of vaginal adenosis, who was also diagnosed with early breast cancer. PMID- 16216746 TI - Fine needle aspiration cytology as an adjunct to core biopsy in the assessment of symptomatic breast carcinoma. AB - INTRODUCTION: Core biopsy (CB) has now largely replaced fine needle aspiration cytology (FNAC) in the pre-operative assessment of breast cancer. We studied the contribution of FNAC, when done as an adjunct to CB, in cancer patients presenting symptomatically. PATIENTS: 112 patients had both CB and FNAC pre operatively and subsequently had surgery (wide local excision or mastectomy). CB & FNAC were done clinically in half the patients and under ultrasound guidance in the majority of others. RESULTS: The complete sensitivity of FNAC was 90% and that of CB was 99%. Absolute sensitivity was 67% and 94%, respectively. CB was not suspicious of malignancy (B1) only in 1 patient in whom FNAC was suspicious (C3). In this patient there was a high degree of suspicion of breast cancer clinically & radiologically so a negative CB would not have been accepted even in the absence of FNAC. CONCLUSIONS: FNAC, when performed in addition to CB, does not provide useful additional information in symptomatic breast cancer patients. PMID- 16216747 TI - An investigation into the effect of axillary surgery on ipsilateral axillary sweating--a pleasing 'complication' or a potential treatment for hyperhidrosis? AB - Although there had not been complaints of excessive sweating pre-operatively, it was noticed incidentally that a large proportion of patients who had undergone axillary lymph node sampling or clearance for breast carcinoma seemed to report ipsilateral sweat loss as a pleasing 'complication'. We interviewed 65 consecutive patients who had all had axillary dissections performed by the same surgeon's technique; the interviews were held 2-102 months after their operations. In 36 cases, patients reported no change in axillary sweating (mean 35 months), whereas 29 patients reported a decrease (mean 50 months). This result was statistically significant at the 95% confidence interval (p=0.022). PMID- 16216748 TI - Invasive ductal carcinoma of the breast with large central acellular zones (ring carcinoma): imaging and clinical findings in eight cases. AB - OBJECTIVE: To review the mammographic and ultrasound appearances in patients who have invasive ductal carcinoma with a central acellular zone (ring carcinoma), as this feature has been reported to be associated with a poorer outcome. MATERIALS: Eight patients were identified with ring carcinomas. Two breast radiologists reviewed their mammograms and ultrasound images. Patient records were reviewed to assess outcome. RESULTS: All patients had lesions deep within the breast, adjacent to the chest wall, five lesions were incompletely visualised on mammography. The appearance was of a circumscribed or obscured mass, without microcalcification. Five patients had ultrasound demonstrating a solid well circumscribed hypoechoic microlobulated lesion. CONCLUSION: In our series of patients who have a ring carcinoma of the breast, mammographic and ultrasound appearances were similar in all cases and lacked the typical features of malignancy. PMID- 16216749 TI - Rationale for the use of trastuzumab in patients with cerebral metastases who previously receive trastuzumab-based therapy for metastatic breast cancer. PMID- 16216750 TI - Defibrillation efficacy testing: long-term follow-up and mortality. AB - AIMS: Extensive defibrillation threshold testing is no longer necessary to perform as devices have become more effective. We assessed the lowest effective defibrillation (LED) level at implantation and before hospital discharge and related this to outcome. METHODS AND RESULTS: One hundred and twenty-seven consecutive patients with biphasic shock and active can devices were studied at intraoperative and predischarge testing. A subgroup of 67 patients had > or = 3 VF inductions at implant. Improvement was defined when LED decreased by > or = 3 J. The LED was significantly higher at implantation compared with predischarge (P < 0.001). Improvement was seen in 73/127 patients (58%). In the group with > or = 3 VF inductions, an implantation LED > 9 J was related to a lower LVEF (P < 0.01); 34/67 patients (51%) had improvement in LED. During follow-up, 18/127 patients died, four received heart transplantation. No different outcome was observed in patients with and without improvement. However, for those with > or = 3 VF inductions, an independent predictor of mortality was implantation LED > 9 J without improvement on the second test. Safety margin < 10 J was not related to mortality. CONCLUSION: Repeated defibrillation efficacy testing before hospital discharge may confirm that a relatively high defibrillation energy is required. This is related to a higher mortality in long-term follow-up. PMID- 16216751 TI - Electroanatomic mapping characteristics of ventricular tachycardia in patients with arrhythmogenic right ventricular cardiomyopathy/dysplasia. AB - BACKGROUND: Ventricular tachycardia (VT) in arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVD) has been previously explored using entrainment mapping techniques but little is know about VT mechanisms and the characteristics of their circuits using an electroanatomical mapping system. METHODS AND RESULTS: Three-dimensional electroanatomical mapping was performed in 11 patients with well tolerated sustained VT and ARVD. Sinus rhythm mapping of the right ventricle was performed in eight patients showing areas of low bipolar electrogram voltage (<1.2 mV). In total 12 tachycardias (mean cycle length 382+/-62 ms) were induced and mapped. Complete maps demonstrated a reentry mechanism in eight VTs and a focal activation pattern in four VTs. The reentrant circuits were localized around the tricuspid annulus (five VTs), around the right ventricular outflow tract (one VT) and on the RV free lateral wall (two VTs). The critical isthmus of each peritricuspid circuit was bounded by the tricuspid annulus with a low voltage area close to it. The isthmus of tachycardia originating from the right ventricular outflow tract (RVOT) was delineated by the tricuspid annulus with a low voltage area localized on the posterior wall of the RVOT. Each right ventricular free wall circuit showed an isthmus delineated by two parallel lines of block. Focal tachycardias originated on the right ventricular free wall. Linear radiofrequency ablation performed across the critical isthmus was successful in seven of eight reentrant tachycardias. The focal VTs were successfully ablated in 50% of cases. During a follow-up of 9-50 months VT recurred in four of eight initially successfully ablated VTs. CONCLUSIONS: Peritricuspid ventricular reentry is a frequent mechanism of VT in patients with ARVD which can be identified by detailed 3D electroanatomical mapping. This novel form of mapping is valuable in identifying VT mechanisms and in guiding RF ablation in patients with ARVD. PMID- 16216753 TI - One heart, two minds. AB - A case of complex atrial tachyarrhythmias following orthotopic heart transplantation is presented. The ablation strategy in such a situation and the outcome achieved are discussed. PMID- 16216752 TI - Indication of the radiofrequency induced lesion size by pre-ablation measurements. AB - BACKGROUND: During radiofrequency ablation of arrhythmias tissue heating and hence lesion size depend on electrode-tissue contact and cooling of the electrode tip caused by cavitary blood flow. These factors are unique and unknown for each catheter placement in the beating heart. A tool for assessing these factors prior to ablation may indicate the lesion size which will be obtained for any given catheter position. METHODS AND RESULTS: Radiofrequency ablation was performed in vitro on strips of left ventricular porcine myocardium during two different levels of convective cooling (0 or 0.1 m/s), two different contact pressures (10 or 30 g) and parallel or perpendicular electrode-tissue orientation using 7F 4 mm tip catheters. Prior to ablation the impedance rise (DeltaIMP) caused by the obtained contact and the temperature rise with a 0.6 W 5 s test pulse (DeltaT) were measured. Subsequently, during unchanged conditions, radiofrequency ablation was performed as either temperature-controlled, power-controlled or irrigated tip ablation and lesion size was determined. DeltaIMP increased significantly (P < 0.05) by improved contact, whereas it was not affected by convective cooling. DeltaT was significantly increased by increasing contact pressure (P < 0.05) and significantly decreased by increased cooling (P < 0.001). DeltaT was not systematically affected by electrode orientation. The product of DeltaT and DeltaIMP showed a significant correlation between the obtained lesion size and power output for temperature-controlled and between lesion size and tip temperature for power-controlled ablation (P < 0.001). CONCLUSIONS: Pre-ablation measurement of DeltaIMP and DeltaT can indicate the lesion size resulting after ablation in temperature-controlled, power-controlled and irrigated ablation in vitro, since DeltaT reflects cavitary cooling and to a smaller extent electrode tissue contact, and DeltaIMP reflects only electrode-tissue contact. PMID- 16216754 TI - Cibenzoline induced Brugada ECG pattern. AB - We report a case of a 61-year-old female patient who presented with palpitations. The baseline electrocardiogram showed incomplete right bundle branch block with saddle back pattern of the ST segment in one precordial lead, but without any significant ST elevation. She was treated with oral cibenzoline. The subsequent ECG showed a coved Brugada ECG (type I) pattern, which resolved following the discontinuation of cibenzoline. An ajmaline test reproduced the coved type Brugada ECG pattern. Our case is the first report of oral cibenzoline therapy unmasking the diagnostic coved Brugada ECG pattern in a patient with a baseline normal ECG. Cibenzoline, a class I sodium channel blocker antiarrhythmic drug, should probably be avoided in the treatment of patients with Brugada syndrome. PMID- 16216755 TI - Further evidence of a close anatomical relation between the oesophagus and pulmonary veins. AB - BACKGROUND: Atrio-oesophageal fistula has been reported as a rare but life threatening complication of ablation of atrial fibrillation (AF). Therefore, the position of the oesophagus in relation to the left atrium (LA) is of major importance for AF ablation. METHODS AND RESULTS: In order to investigate the possible anatomical variability between the oesophagus and the left atrium, multidetector-row spiral computed tomography (MDCT) of 60 healthy males (age 58.1+/-5.1 years; LA diameter 5.4+/-0.7 x 3.8+/-0.6 cm; LA volume 60.5+/-15.4 ml) was analyzed. The distance between the oesophagus and the ostia of the pulmonary veins (PV) ranged between 0 and 50.7 mm. Especially for the left PV, the oesophagus was closer than 5 mm to the ostia in 29 cases (48%; n = 24 for left superior PV; n = 10 for left inferior PV; n = 0 for right superior PV; n = 1 for right inferior PV). In addition, the oesophagus was very close to the LA wall (0.8+/-0.9 mm; range 0-3.3 mm). Intraobserver variability was 1.1+/-0.7 mm or 3.5%. CONCLUSION: The position of the oesophagus in relation to the LA and the PV demonstrates high variability. In many cases, the oesophagus is very close to the ostia of the PVs and lies only a short distance from the LA wall. Thus, an anatomical localization of the oesophagus may be critical before or during AF ablation to prevent atrio-oesophageal fistula, especially as there is a need for transmural atrial lesions. PMID- 16216756 TI - Comparison of acute and long-term effects of single-dose amiodarone and verapamil for the treatment of immediate recurrences of atrial fibrillation after transthoracic cardioversion. AB - AIMS: Amiodarone and verapamil have been employed to treat immediate recurrences of AF (IRAF) after cardioversion. This study compares the efficacy of these agents for the treatment of IRAF. METHODS AND RESULTS: One hundred and eighty five patients underwent transthoracic cardioversion (CV) for AF. AF recurred within 10 min in 20 patients (10.8%). These patients were randomized to verapamil (seven patients), or amiodarone (13 patients). After administration of verapamil and repeat CV, five patients (71%) experienced IRAF, compared with seven patients (54%) receiving amiodarone (P = 0.4). Including the results after crossover, IRAF occurred in 8/10 patients (80%) who received verapamil, compared with 7/15 patients (47%) who received amiodarone (P = 0.1). The combination of these agents prevented IRAF in 10/20 patients (50%). After a follow-up of 319+/-189 days, 42% of the IRAF patients treated with verapamil and/or amiodarone remained in sinus rhythm, which did not differ from patients without IRAF (53%, P = 0.7). CONCLUSIONS: IRAF occurs in 10% of patients undergoing CV. Amiodarone and verapamil are effective in preventing IRAF and result in a sinus rhythm maintenance rate of 50%. Since there is no difference in the long-term maintenance of sinus rhythm between patients with and without IRAF, attempts to restore sinus rhythm after pharmacological pretreatment are justified. PMID- 16216757 TI - Does treatment with ACE inhibitors or angiotensin II receptor antagonists prevent atrial fibrillation after dual chamber pacemaker implantation? AB - AIMS: A retrospective observational study was performed to test the hypothesis that a lower incidence of atrial fibrillation (AF) would be observed in patients treated with either angiotensin converting enzyme (ACE) inhibitors or angiotensin II receptor antagonists (AIIRAs) than those without these drugs, 1-year following implantation of a dual chamber pacemaker for all indications. METHODS: One hundred and sixty consecutive patients who underwent implantation of a dual chamber pacemaker between January and August 2002 were identified and their case notes were retrospectively analysed. The primary endpoint was the presence of persistent AF (confirmed by 12-lead ECG recorded from the visit to the pacemaker clinic) at 12-month follow-up. RESULTS: Overall, 8% patients developed new onset persistent AF at 1-year follow-up. The incidence of AF at 1-year was 4% in patients treated with ACE inhibitors, 8% in patients taking AIIRAs or 5% on either drug. Although a trend towards a higher incidence of AF was observed at 1 year (10%) in patients not receiving either of these drugs, this was not statistically significant (P = 0.21, drug vs. no drug). The incidence of AF in patients with a previous history of paroxysmal AF or cardioversion was significantly higher (23%) than those patients without (5%), P < 0.0001. An odds ratio (95% CI) of 7.9 (2.3-27.8) was obtained. CONCLUSION: To confirm these interesting initial results and to investigate this important relationship further, larger prospective randomised controlled studies are required. PMID- 16216758 TI - The effects of 5-HT4 receptor blockade and stimulation, during six hours of atrial fibrillation. AB - AIMS: Stimulation of atrial 5-HT4 receptors is associated with arrhythmias. Their blockade prolongs atrial effective refractory period (ERP), following short runs of atrial fibrillation (AF). The role of 5-HT4 receptors during longer periods of AF is unknown. In this study, we investigated the effects of the selective 5-HT4 receptor stimulation and blockade on porcine atria, during 6 h of AF. METHODS: Atrial ERP, monophasic action potential (MAP) duration, time to sinus rhythm restoration (TSRR) and ERP/MAP ratio were assessed in 27 pigs, at baseline and every hour, during 6 h of AF, induced by rapid atrial pacing. Ten animals were used as controls, 10 were administered the selective 5-HT4 antagonist SB203186 and seven were administered the selective 5-HT4 agonist RS67333. RESULTS: During the first few hours of fibrillation, ERP, MAP and TSRR were preserved in SB203186 treated pigs, while they were shortened in controls and RS67333-treated animals. After 6 h of arrhythmia, ERP and MAP were shortened in all three groups, but the decrease was less in SB203186-treated pigs. ERP/MAP ratio increased in controls and RS67333-treated animals, while it remained unchanged in SB203186-treated pigs. Towards the end of the AF period, four of the SB203186-treated pigs developed sustained atrial tachycardia. CONCLUSION: Following short periods of AF, 5-HT4 receptors' blockade protects the porcine atria against ERP and MAP shortening, while their stimulation has the opposite result. This beneficial effect, though, is gradually diminished following longer periods of AF and atrial tachycardia may develop. PMID- 16216759 TI - Long-term results of endocardial pacing with Autocapture threshold tracking pacemakers in children. AB - AIM: We aimed to evaluate the long-term results of endocardial pacing with Autocapture threshold tracking pacemakers in children. METHODS AND RESULTS: Implantation and follow-up data of 20 children with these pacemakers were retrospectively evaluated. The pacemakers were implanted subpectorally in five and subcutaneously in 15 patients. The indication for pacing was high-grade atrioventricular block in 18 cases. The mean age at implantation was 7+/-4.8 years. Four patients were pacemaker dependant (heart rate < 30 bpm). At implantation, the mean pacing threshold was 0.5 V at 0.5 ms. The mean evoked response (ER) signal was 8.5+/-3.6 mV, and the polarisation signal (PS) was <1 mV in 15 patients and 1-2 mV in five patients. During the mean follow-up period of 60 months, mean ER signal decreased significantly to 7.7+/-6.3 mV at 24 months and 6.5+/-2.5 mV at 60 months (P < 0.05). In four of 15 patients (26.6%), with a predischarge PS value of <1 mV, it increased between 1 and 2 mV over time. During follow-up, autocapture function was deactivated in six (30%) patients; due to inappropriate ER/PS values in four and due to severe muscle twitching in two with subpectoral implants. These problems occurred during a median period of 21 months after implantation. Generators were replaced in three patients with Microny pacemakers because of battery depletion at 54, 66 and 78 months. In two of them autocapture function had been working since implantation. In seven of 10 patients, who completed > or = 60 months of follow-up, battery impedances were still at the predischarge level. CONCLUSIONS: Autocapture function works well in most children at implantation. Mean ER signal significantly decreases over time despite stable pacing parameters. Autocapture function may become nonoperational due to decreased ER signal in some patients. Muscle twitching may be an important problem that may result in discontinuation of autocapture function in children with subpectoral implants. PMID- 16216760 TI - Effects of variation of atrioventricular interval on left ventricular diastolic filling dynamics and atrial natriuretic peptide levels in patients with DDD pacing for complete heart block. AB - AIMS: This study was designed to evaluate the effect of variation of atrioventricular (AV) interval (AVI) on left ventricular (LV) diastolic function and ANP and c-GMP levels during DDD pacing in patients with complete AV block and normal systolic function. METHODS: The study population comprised 22 patients (mean age 65.2+/-14.3, 12 males) with complete AV block. All patients underwent complete Doppler echocardiography before implantation of a DDD-pacemaker. Twenty four hours later, patients were paced for a period of 30 min, at three different AVIs (100 ms, 150 ms and 200 ms), at rest. During each pacing period, Doppler derived LV diastolic indices were re-evaluated and ANP and c-GMP levels were reassessed. RESULTS: Overall comparison showed a significant progressive augmentation, from 200 ms to 100 ms AVI, in transmitral E/A wave ratio (from 0.53+/-0.13 to 0.90+/-0.25, P = 0.0005) and in LV filling time (from 0.33+/-0.05 to 0.40+/-0.06s, P = 0.0005), followed by a significant progressive reduction in ANP and c-GMP levels. An AVI of 100 ms or 150 ms was associated with improved diastolic indices and lower natriuretic peptides levels, compared with the longer AVI. CONCLUSION: Programmed AVI during DDD pacing affects LV diastolic performance and plasma ANP and c-GMP levels. The assessment of these parameters constitutes a useful modality for AVI optimization. PMID- 16216761 TI - Effects of right ventricular pacing on regional myocardial glucose metabolism. AB - AIMS: Permanent right ventricular apical pacing (RVP) is associated with a wide range of myocardial abnormalities. The purpose of this study was to determine the changes over time of RVP on myocardial blood flow (MBF) and glucose metabolism as assessed by positron emission tomography (PET). METHODS: In eight candidates for permanent pacemaker implantation PET imaging was performed with 13N-ammonia and 18F-Fluorodeoxyglucose (FDG) to assess MBF and glucose metabolism before (PET1) and repeated after 3 months of RVP (PET2). For the analysis, the left ventricle was divided into three parts (apex, mid-ventricular and base) and subdivided into six segments (inferior, posterior, lateral, anterior, antero-septal and infero septal). RESULTS: After RVP, defects of FDG uptake were found in the left ventricle near the stimulation site, without corresponding changes in MBF. Changes over time in the mean FDG uptake were statistically significant between PET1 and PET2 in the apical inferior, apical-posterior, apical-anterior, apical antero-septal, apical infero-septal, mid-inferior and mid-infero-septal segments. CONCLUSIONS: This study shows that RVP induces major changes in the distribution of FDG uptake in the left ventricular myocardium. FDG uptake significantly decreases in the regions surrounding the pacing site. PMID- 16216762 TI - Runaway pacemaker: a forgotten phenomenon? AB - Runaway is an uncommon pacemaker dysfunction, characterized by fast and erratic spikes at non-physiological rates. This infrequent but potentially lethal failure mode may be related to low battery voltage. Four single chamber pacemaker patients were analyzed (Medtronic Minix ST 8330, Minneapolis, MN, had been implanted in two patients and two CPI Triumph VR 1124, St Paul, MN, in the other two). They had been admitted because of presyncopal episodes. Typical high rate stimuli at 2000 ppm alternating with pacing at 60-65 ppm were recorded in all ECGs. Lead system tests were normal. The pulse generators had to be replaced. PMID- 16216763 TI - Twins with implanted pacemakers: Is there an increased mortality risk for the co twin? A follow-up study based on the Danish Twin Registry and the Danish Pacemaker Register. AB - BACKGROUND: The main indications for implanting a cardiac pacemaker are either acquired atrio-ventricular block (AVB) or sick sinus syndrome (SSS). In general, the exact aetiology of these conditions is unknown. Genetic factors may play a role. Untreated AVB is associated with increased mortality but SSS seems to have a good prognosis. The present study was designed to assess whether first-degree relatives of these patients have an increased mortality risk. METHODS: We investigated the survival pattern of the co-twins of twins who have had a cardiac pacemaker implanted, by linking two nationwide registries in Denmark: the Danish Pacemaker Register and the Danish Twin Registry. For each "pacemaker-twin" four control-twins matched with respect to age, sex, and zygosity were selected among the twins who were alive at the time of the pacemaker implantation. A total of 159 twins with a pacemaker treated bradycardia of unknown aetiology were identified of which 73 had AVB and 64 had SSS. A total of 622 matched control twins could be identified. The survival from age six of the co-twins of the "pacemaker-twins" was compared with that of the co-twins of the control-twins. The follow-up period ended January 1st, 2000. FINDINGS: The survival curves for the co-twins of the "pacemaker-twins" were virtually identical to those of the co twins of the control-twins. This finding was consistent for both AVB and SSS and for monozygotic as well as dizygotic twins. CONCLUSION: No increased mortality risk could be demonstrated for first-degree relatives--including monozygotic twins--of patients with a cardiac pacemaker implanted due to either acquired AVB or SSS. PMID- 16216764 TI - Evaluation of left bundle branch block as a reversible cause of non-ischaemic dilated cardiomyopathy with severe heart failure. A new concept of left ventricular dyssynchrony-induced cardiomyopathy. AB - OBJECTIVES: We sought to determine if amelioration of left bundle branch block (LBBB)-induced contraction disturbances achieved by left ventricular (LV)-based pacing could result in sustained reversal of severe LV dysfunction in certain patients with chronic heart failure due to non-ischaemic cardiomyopathy. BACKGROUND: It has been shown that LBBB induces asynchronous contraction of LV. However, whether such a functional contraction disturbance, if present for an extended period of time, could account for a dilated cardiomyopathy remains unknown. METHODS: The study population comprised 29 patients with dilated cardiomyopathy, sinus rhythm, LBBB and severe heart failure (14 patients in New York Heart Association (NYHA) class III and 15 in class IV). Patients were followed prospectively after resynchronization therapy. LV function was considered to be normalized when ejection fraction (EF) was >50% at 1 year. RESULTS: Five among the 29 patients (17%: group 1) demonstrated both complete normalization of LV function following resynchronization therapy (EF: from 19+/-6 to 55+/-3%, P = 0.001) and clinical improvement (mean NYHA class: 3.4+/-0.5 to 1.8+/-0.4, P = 0.02; 6-min walk distance: 300+/-136 to 444+/-75 m, P = 0.12; peak VO2: 11.9+/-4 to 15.8+/-2 ml/min/kg, p = 0.03). Among the remaining 24 patients (83%: group 2) EF improved but did not normalize (from 21+/-8 to 23+/-11%, ns). Baseline clinical features could not predict which patients would exhibit the reversal of LV dysfunction. CONCLUSIONS: Normalization of LV function 1 year after resynchronization therapy in a small but important number of patients suggests that long-standing LBBB may be a newly identified reversible cause of cardiomyopathy. PMID- 16216765 TI - Left atrial far-field sensing by left ventricular leads: a potential hazard in cardiac resynchronisation therapy. AB - BACKGROUND: Cardiac resynchronisation therapy (CRT) requires a lead advanced through the coronary sinus (CS) to pace the left ventricle (LV). Left atrial far field signals (LAFFS) may be sensed by the LV lead at the time of implant or after lead dislodgement, and may inhibit ventricular pacing. OBJECTIVE: To assess the incidence of detection of LAFFS > 2 mV and its correlation with the CS lead position. METHODS: Data from the first 75 consecutive patients enrolled in the InSync III multicentre study were analysed. The position of the LV lead was recorded at implant. During follow-up, pacing was temporarily inhibited and the LV channel electrogram was recorded. The amplitude of LAFFS observed before discharge from the hospital and at 1 month of follow-up was retrospectively analysed. A LAFFS > 2 mV was considered clinically significant. RESULTS: CRT systems were successfully implanted in 71 of 75 patients. A LAFFS > 2 mV was recorded by the LV lead channel in six of 71 patients (8.5%). This phenomenon developed between hospital discharge and 1 month of follow-up in two of these patients and in one case disappeared within 1 month. It was observed in all CS tributaries except the anterior and mid-cardiac veins. CONCLUSIONS: Left atrial far-field signals sensed by the LV lead were not rare. Implanting physicians should be aware of this phenomenon in order to prevent potentially serious complications. PMID- 16216766 TI - Transoesophageal left ventricular pacing in heart failure patients with permanent right ventricular pacing. AB - BACKGROUND: Previous studies of biventricular (BV) pacing for treatment of heart failure (HF) patients with left bundle branch block (LBBB) evaluated responders to BV pacing with acute transvenous left ventricular (LV) pacing and arterial pulse pressure (PP). The aim of this study was to assess transoesophageal LV pacing in evaluation of the haemodynamic response with a view to upgrading responders from permanent right ventricular (RV) pacing to BV pacing. METHODS AND RESULTS: Ten HF patients (age 62+/-8 years; one female, nine males) in NYHA III, LV ejection fraction 24+/-9% and permanent RV pacing by means of an implanted pacemaker or ICD were tested using transoesophageal LV pacing and PP. Permanently RV-paced HF patients were analysed with transoesophageal atrial sensed LV pacing in VAT mode with a different AV delay (n = 6) and with transoesophageal LV pacing in V00 mode during atrial fibrillation (n = 4). In five responders, PP was higher during transoesophageal LV pacing than PP during RV pacing (74+/-42 versus 57+/ 31 mmHg, P = 0.015). Responders were upgraded by means of an LV lead via the coronary sinus in the posterior (n = 1) or posterolateral (n = 4) walls and after attaining a high LV pacing threshold with an epicardial LV lead on the anterior (n = 1) or anterolateral (n = 1) walls. NYHA class improved from 3 to 2+/-0.3 (P = 0.003) during 204+/-120 days follow-up and cardiac output increased from 4.4+/ 1.5 to 5.6+/-1.7 l/min (P = 0.027) when comparing BV pacing and optimal AV delay with RV pacing. In five nonresponders, PP was not higher during transoesophageal LV pacing than during RV pacing. CONCLUSION: Transoesophageal LV pacing may be a useful technique to detect responders to BV pacing in permanently RV-paced HF patients. PMID- 16216767 TI - Paradoxical effect of isoprenaline infusion. AB - BACKGROUND: Isoprenaline (isoproterenol) is a beta-adrenergic drug, used to increase the heart rate and, during electrophysiological study, to facilitate the induction of supraventricular (SVT) and ventricular tachycardias (VT). Decrease in heart rate during isoprenaline infusion is a rare phenomenon. The purpose of the study was to evaluate the incidence, the possible mechanisms, and the significance of a paradoxical bradycardia induced by isoprenaline infusion. METHODS: Electrophysiological study was performed for the evaluation of tachycardias (n = 551) or dizziness/syncope (n = 214) in 765 patients aged from 15 to 85 years. The study was negative in the basal state, and was repeated after isoprenaline infusion (2-4 microg/min). RESULTS: In 714 patients, this perfusion increased the heart rate to 100-140 bpm. A bradycardia was noted in 51 patients (7%). Two bradyarrhythmias were noted: (1) sinus or junctional bradycardia (cycle length - 10%) occurred in 43 patients, aged 15-70 years, generally studied for unexplained syncope (n = 37); a young age (40+/-16 years), syncope (n = 37) and absence of heart disease (n = 27) were more frequent than that in patients without isoprenaline-induced sinus or junctional bradycardia; another arrhythmia (SVT or VT) was induced in seven patients with syncope, five with heart disease and two without; six young patients (<50 years) had no syncope and were studied for SVT or VT; (2) eight patients, aged 65+/-11 years, developed second-degree atrioventricular (AV) block which was supraHisian (n = 4) or infraHisian (n = 4); they were studied for exercise-related syncope; they had no signs of myocardial ischaemia and AV block was reproduced by ajmaline testing: isoprenaline revealed organic conduction disturbance. CONCLUSION: The occurrence of paradoxical bradycardia was a rare finding during isoprenaline infusion (7%); sinus or junctional bradycardia was a sign of hypervagotonia, but was without clinical significance in 35% of these patients. The development of second-degree AV block was always pathological and associated with AV conduction disturbances, which occurred spontaneously during exercise. Isoprenaline infusion appeared to be a simple means to detect organic AV conduction disturbance in patients complaining of exercise or stress-related dizziness/syncope and unable to perform exercise test. PMID- 16216768 TI - Repeated tilt testing in patients with tilt-positive neurally mediated syncope. AB - In this study we have included 222 patients with apparent neurally mediated syncope and with a positive diagnostic tilt test. The mean age was 33.4+/-21.2 years (median 25.3): there were 107 men (median age 25.3) and 115 women (median age 22.6). The age difference between males and females was statistically significant (P = 0.002). The response to the diagnostic tilt test was: type 1 (mixed) in 74 patients; type 2A (cardioinhibitory and bradycardia) in 6; type 2B (cardioinhibitory and asystole) in 61; type 3 (vasodepressor) in 81. In all 222 patients the diagnostic tilt test was positive after 19+/-11 min (mean+/-SD), median time: 18 min. For the four types of syncope, the duration in minutes of the diagnostic tilt test was: type 1 (mixed) 19.5+/-11.4; type 2A (cardioinhibitory) 24.8+/-13.6; type 2B (cardioinhibitory and asystole) 14.7+/ 10.2; type 3 (vasodepressor) 21.6+/-11.1. A significant difference was found between type 2B and type 3 responses (P = 0.002). Between males and females no significant differences in the duration of the diagnostic tilt test were found, neither for all responses, nor for the four subtypes. A type 2B (cardioinhibitory and asystole) response occurred in 61 patients. The duration of asystole was 12.8+/-10.6s (mean+/-SD; median 9, minimum 3, maximum 60). The head-up tilt test was repeated day after day: one session per day. The response became negative at the second session in 119 patients (54%); at session 3 in 47 (21%); at session 4 in 30 (13%); at session 5 in 15 (7%); at session 6 in 6 (3%); at session 7 in 2 (1%); at session 8 in 3 (1%). For all 222 patients the mean number of sessions in order to obtain a negative tilt test was 2.9 (SD 1.3; median 2). Only 25% of patients remained tilt-positive for three or more sessions. A negative tilt test was ultimately obtained in every patient. Follow-up data are available for 202/222 patients. The time span between the first and last tilt test was 11.1+/ 10 months (median 8.8). Of these 202 patients, 163 remained free of any event (80.7%). PMID- 16216770 TI - Ventricular fibrillation induced by carotid sinus massage without preceding bradycardia. AB - Carotid sinus massage is widely used to detect carotid sinus hypersensitivity in patients presenting with syncope. Although generally safe, the risks associated with the procedure may not be fully appreciated by either the patient or the attending medical staff. We present the case of a patient who developed ventricular fibrillation during carotid sinus massage, not explained by preceding bradycardia or concomitant predisposing heart disease, and which highlights the need for ready availability of resuscitation equipment during this procedure. PMID- 16216769 TI - Distribution of syncopal episodes in children and adolescents with neurally mediated cardiac syncope through the day. AB - AIMS: To assess the relation between the timing of syncopal attacks and tilt test positivity. METHOD AND RESULTS: Prospective comparisons of distribution of syncopal attacks in 49 consecutive neurally mediated cardiac syncope (NMCS) patients (19 boys, 30 girls, mean age 13.7+/-0.68) were evaluated. Head-up tilt test (HUT) was positive in 28 patients and negative in 21. A questionnaire was given to every patient about the time and number of the syncopal attacks, presyncopal symptoms or signs before HUT. Although syncopal attacks were found to be concentrated in the morning especially between 10 AM and 12 noon in HUT positive patients (P < 0.001), there was a concentration in the late afternoon and evening period of the day especially between 2 PM and 6 PM in HUT negative patients (P < 0.001). CONCLUSION: It was assumed that diurnal variation in autonomic function may be the factor in the timing of syncopal events during morning hours in the HUT positive NMCS children and adolescents. PMID- 16216773 TI - Towards quantitative biology: integration of biological information to elucidate disease pathways and to guide drug discovery. AB - Developing a new drug is a tedious and expensive undertaking. The recently developed high-throughput experimental technologies, summarised by the terms genomics, transcriptomics, proteomics and metabolomics provide for the first time ever the means to comprehensively monitor the molecular level of disease processes. The "-omics" technologies facilitate the systematic characterisation of a drug target's physiology, thereby helping to reduce the typically high attrition rates in discovery projects, and improving the overall efficiency of pharmaceutical research processes. Currently, the bottleneck for taking full advantage of the new experimental technologies are the rapidly growing volumes of automatically produced biological data. A lack of scalable database systems and computational tools for target discovery has been recognised as a major hurdle. In this review, an overview will be given on recent progress in computational biology that has an impact on drug discovery applications. The focus will be on novel in silico methods to reconstruct regulatory networks, signalling cascades, and metabolic pathways, with an emphasis on comparative genomics and microarray based approaches. Promising methods, such as the mathematical simulation of pathway dynamics are discussed in the context of applications in discovery projects. The review concludes by exemplifying concrete data-driven studies in pharmaceutical research that demonstrate the value of integrated computational systems for drug target identification and validation, screening assay development, as well as drug candidate efficacy and toxicity evaluations. PMID- 16216774 TI - The duckweeds: a valuable plant for biomanufacturing. AB - Inherent characteristics of duckweed, including fast, clonal growth, small size and simple growth habit, argue for their use as a biomanufacturing platform for proteins, polymers and small molecules. This review addresses five areas relevant to commercialization of the duckweed platform: (1) the characteristics of wild type duckweed and general cultural requirements; (2) the genetics and biochemistry of the plants and recent scientific developments that provide the technology necessary to genetically modify duckweed; (3) the advantages provided by inherent duckweed characteristics and genetic engineering technology relative to bioproduction; (4) recent progress towards commercialization of duckweed-based products and (5) the major research needs for further R&D. PMID- 16216775 TI - The application of BacMam technology in nuclear receptor drug discovery. AB - The nuclear receptor (NR) superfamily represents a major class of drug targets for the pharmaceutical industry. Strategies for the development of novel, more selective and safer compounds aimed at these receptors are now emerging. Reporter assays have been used routinely for the identification and characterisation of NR ligands. As the NR drug development process evolves, the increase in screening demand in terms of both capacity and complexity has necessitated the development of novel assay formats with increased throughput and flexibility. BacMam technology, a modified baculovirus system for over-expressing genes of interest in mammalian cells has helped answer this requirement. BacMam has many advantages over traditional gene delivery systems including high transduction efficiencies, broad cell host range, speed, cost and ease of generation and use. As outlined in this review, the technology has shown itself to be robust and efficient in various NR assay formats including transactivation (ER alpha/beta, MR, PR and PXR) and transrepression (GR-NFkappaB). In addition, the flexibility of this system will allow greater multiplexing of receptor, reporter, and cell host combinations as NR assays become more complex in order to relate better to relevant cellular and biological systems. PMID- 16216776 TI - Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. AB - Tetrazolium salts have become some of the most widely used tools in cell biology for measuring the metabolic activity of cells ranging from mammalian to microbial origin. With mammalian cells, fractionation studies indicate that the reduced pyridine nucleotide cofactor, NADH, is responsible for most MTT reduction and this is supported by studies with whole cells. MTT reduction is associated not only with mitochondria, but also with the cytoplasm and with non-mitochondrial membranes including the endosome/lysosome compartment and the plasma membrane. The net positive charge on tetrazolium salts like MTT and NBT appears to be the predominant factor involved in their cellular uptake via the plasma membrane potential. However, second generation tetrazolium dyes that form water-soluble formazans and require an intermediate electron acceptor for reduction (XTT, WST-1 and to some extent, MTS), are characterised by a net negative charge and are therefore largely cell-impermeable. Considerable evidence indicates that their reduction occurs at the cell surface, or at the level of the plasma membrane via trans-plasma membrane electron transport. The implications of these new findings are discussed in terms of the use of tetrazolium dyes as indicators of cell metabolism and their applications in cell biology. PMID- 16216777 TI - Display libraries on bacteriophage lambda capsid. AB - Phage display is an established technology that has been successfully applied, in the last fifteen years, to projects aimed at deciphering biological processes and/or at the isolation of molecules of practical value in several diverse applications. Bacteriophage lambda, representing a molecular cloning and expression tool widely utilized since decades, has also been exploited to develop vectors for the display of libraries on its capsid. In the last few years, lambda display approach has been consistently offering new enthralling perspectives of technological application, such as domain mapping, antigen discovery, and protein interaction studies or, more generally, in functional genomics. PMID- 16216778 TI - Vibrational spectroscopy for molecular characterisation and diagnosis of benign, premalignant and malignant skin tumours. AB - Understanding the molecular, cellular and tissue changes that occur during skin carcinogenesis is central to cancer research in dermatology. The translational aspects of this field--the development of clinical applications in dermatology from the laboratory findings--aim at improving clinical diagnosis, monitoring and treatment of skin cancer. Vibrational spectroscopy, both infrared (IR) and Raman spectroscopy, would be helpful in achieving those goals, since it has been shown to have potential in characterising and discriminating tumour and dysplastic tissue from normal tissue. Clinically differential diagnosis of skin tumours is often difficult and a histopathologic analysis of skin biopsies remains the standard for diagnostic confirmation. We review and update the literature on the subject, demonstrating that the IR and Raman spectra of skin tissues provide valid and useful diagnostic information about a number of skin tumours. We also include a survey of introduced sampling methods for IR and Raman spectroscopy in dermatology, and additionally describe the differences between microscopic, macroscopic and fibreoptic diagnosis of skin cancer. Although in its early stages, we remain optimistic that vibrational spectroscopy has the potential to be fully accepted as a rapid screening tool with sufficient sensitivity and specificity for non-destructive in vitro, ex vivo and in vivo analyses by the dermatological community. Further progress toward molecular characterisation of skin cancer by vibrational spectroscopy would have important research and clinical benefits in dermatology. PMID- 16216779 TI - Cell and tissue autofluorescence research and diagnostic applications. AB - Cells contain molecules, which become fluorescent when excited by UV/Vis radiation of suitable wavelength. This fluorescence emission, arising from endogenous fluorophores, is an intrinsic property of cells and is called auto fluorescence to be distinguished from fluorescent signals obtained by adding exogenous markers. The majority of cell auto-fluorescence originates from mitochondria and lysosomes. Together with aromatic amino acids and lipo-pigments, the most important endogenous fluorophores are pyridinic (NADPH) and flavin coenzymes. In tissues, the extracellular matrix often contributes to the auto fluorescence emission more than the cellular component, because collagen and elastin have, among the endogenous fluorophores, a relatively high quantum yield. Changes occurring in the cell and tissue state during physiological and/or pathological processes result in modifications of the amount and distribution of endogenous fluorophores and chemical-physical properties of their microenvironment. Therefore, analytical techniques based on auto-fluorescence monitoring can be utilized in order to obtain information about morphological and physiological state of cells and tissues. Moreover, auto-fluorescence analysis can be performed in real time because it does not require any treatment of fixing or staining of the specimens. In the past few years spectroscopic and imaging techniques have been developed for many different applications both in basic research and diagnostics. PMID- 16216780 TI - Sterilisation in biotechnology. AB - Sterilisation of fermentation media, inlet and outlet gases and maintenance of aseptic conditions during bioprocessing represents one of the most relevant parts of comprehensive upstream processing. The present chapter reviews various sterilisation techniques and methods for sterilisation of inlet gases, liquids, including sterilisation of small and large industrial equipment and validation of sterilisation procedures. Various methods for sterilisation using different kinds of filters, filter cartridges and membrane filters are also presented. Liquid sterilisation methods by filtration as well as heat sterilisation are discussed. Sterilisation of small and industrial scale equipment by chemical agents and physical methods as ionising radiation and dry heat sterilisation including the validation of sterilisation are discussed. PMID- 16216781 TI - Convective Interaction Media (CIM)--short layer monolithic chromatographic stationary phases. AB - Modern downstream processing requires fast and highly effective methods to obtain large quantities of highly pure substances. Commonly applied method for this purpose is chromatography. However, its main drawback is its throughput since purification, especially of large molecules, requires long process time. To overcome this problem several new stationary phases were introduced, among which short layer monoliths show superior properties for many applications. The purpose of this review is to give an overview about short methacrylate monolithic columns commercialised under the trademark Convective Interaction Media (CIM). Their unique properties are described from different perspectives, explaining reasons for their application on various areas. Approaches to prepare large volume methacrylate monolithic column are discussed and optimal solutions are given. Different examples of CIM monolithic column implementation are summarised in the last part of the article to give the reader an idea about their advantages. PMID- 16216782 TI - Detection of metazoan species as a public health issue: simple methods for the validation of food safety and quality. AB - Species identification represents a critical issue in food chain safety and quality control. Several procedures are available to detect animal proteins in cattle feed or to trace transgenic foods. The most effective approach is based on the use of DNA as a marker. Amplification of DNA provides rapid, sensitive and specific protocols. Several target genes can be used, but new insights come from the mitochondrial genome, which is naturally amplified in each cell and shows a remarkable resistance to degradation. These are key points when analysing complex matrices such as foods, animal feedstuff or environmental samples. Traceability is important to prevent BSE or to monitor novel foods, such as genetically modified organisms. Amplification is commonly performed, but it requires expertise and a molecular biology laboratory to perform restriction analysis, electrophoresis or gel staining for the visualisation of results. Hereby, we consider a strategy based on multiple nested amplification and reverse hybridisation assay that virtually requires only a thermocycler and a water bath. The protocol is rapid and simple and can simultaneously detect different species in a DNA sample. This promising approach allows microarray developments, opening up to further perspectives. An international application has been published under the patent cooperation treaty. Presently, a ban on feeding ruminants on cattle derived proteins is in force in Europe and USA. The identification of metazoan traces in a sample is not only a mere preventive measure for BSE, but represents a possible screening system for monitoring biotechnology products and procedures, as well as a quality control strategy to assure consumer's rights. PMID- 16216783 TI - Water ecology of Legionella and protozoan: environmental and public health perspectives. AB - Ecological studies on Legionella spp. are essential to better understand their sources in the natural environments, the mechanism of their entry into man-made water systems and the factors enabling their survival and growth in aquatic habitats. Legionella spp. exhibits peculiar and multiple strategies to adapt to stressful environment conditions which normally impair other germ survival. These strategies include the ability to enter in a viable but non-cultivable (VBNC) state, to multiply intracellularly within a variety of protozoa, such as amoebae, to survive as free organisms within biofilms and to be enhanced/inhibited by the presence of other aquatic bacteria. The host-parasite interaction has been shown to be central in the pathogenesis and ecology of L. pneumophila. The bacterial protozoan interaction contributes to the amplification of Legionella population in water systems, represents a shelter against unfavourable environmental conditions, acts as a reservoir of infection and contributes to virulence by priming the pathogen to infect human cells. Legionella is able to survive as free organism for long periods within biofilms which are widespread in man-made water systems. Biofilm provides shelter and nutrients, exhibits a remarkable resistance to biocide compounds and chlorination, thus representing ecological niches for legionella persistence in such environments. Further knowledge on biofilm associated legionellae may lead to effective control measures to prevent legionellosis. Lastly, new perspectives in controlling legionella contamination can arise from investigations on aquatic bacteria able to inhibit legionella growth in natural and artificial water systems. PMID- 16216785 TI - Cell migration/invasion assays and their application in cancer drug discovery. AB - Invasive capacity is the single most important trait that distinguishes benign from malignant lesions. Tumour cells, during intravasation and extravasation of blood and lymphatic channels and when establishing colonies at secondary sites, must move through tissue boundaries that normal adult cells (other than, for example activated leukocytes) do not cross. Similar mechanisms are also utilised by activated endothelial cells during the generation of new blood vessels that enable the sustained growth and dissemination of tumours. It is now increasingly recognised that these processes--cell motility and invasion--might provide a rich source of novel targets for cancer therapy and that appropriate inhibitors may restrain both metastasis and neoangiogenesis. This new paradigm demands screening assays that can rapidly and quantitatively measure cell movement and the ability to traverse physiological barriers. We also need to consider whether simple reductionist in vitro approaches can reliably model the complexity of in vivo tumour invasion/neoangiogenesis. There are both opportunities and challenges ahead in developing a balanced portfolio of assays that will be able to evaluate accurately and finally deliver novel anti-invasive agents with therapeutic potential for clinical use. PMID- 16216784 TI - A prospective, non-randomised phase 1-2 trial of VACOP-B with filgrastim support for HIV-related non-Hodgkin's lymphoma. AB - Non-Hodgkin's lymphoma (NHL) remains an important complication of associated HIV infection despite advances in antiretroviral therapy (ART), and the optimum chemotherapy regimen for this disease remains to be defined. A dose-escalation trial was performed to determine the maximum tolerated doses of etoposide and doxorubicin as part of the 12-week VACOP-B regimen, supported by filgrastim (r metHuG-CSF). Patients with aggressive histology HIV-related NHL who were previously untreated with chemotherapy, and who had no active opportunistic infection were eligible for the study. Chemotherapy consisted of cyclophosphamide 350 mg/m2, vincristine 2 mg, bleomycin 10 U/m2; and prednisone 100 mg q2 days x 12 weeks, with increasing doses of doxorubicin 25-50 mg/m2 and etoposide 25-50 mg/m2 intravenously and 50-100 mg/m2 orally. Central nervous system prophylaxis (intrathecal cytarabine 50 mg x 4 doses), antifungal, and Pneumocystis carinii prophylaxis were used, and filgrastim was administered to prevent neutropenic complications. One dose level was expanded to permit the concomitant use of ART. Endpoints were determination of maximum tolerated dose of doxorubicin and etoposide, treatment tolerability, and survival. Forty-seven patients were enrolled, most with diffuse large-cell or immunoblastic NHL. Protocol-defined maximum tolerated dose was not reached and the limits of dose-limiting toxicity were not exceeded, even in patients receiving ART. Thirty-two cycles (4.9%) were delayed >6 days because of toxicity; 30 patients (64%) completed all 12 weeks of treatment. After completion of therapy, 14 patients had a complete response (30%), and 4 had a partial response (8%). Median time to progression was 9 months. At 42 months, progression-free survival was 25% and overall survival was 28%. PMID- 16216788 TI - London 2005, bruised but not broken. PMID- 16216789 TI - The 14th World Congress on Disaster and Emergency Medicine introduces an international nursing section. PMID- 16216792 TI - Nursing students' perceptions about disaster nursing. PMID- 16216793 TI - The role of public health nurses in bioterrorism preparedness. AB - BACKGROUND: Public health nurses have a central function in the public health system. Nurses conduct disease surveillance, which is an important first step in recognizing diseases caused by bioterrorist agents. Unfortunately, the current public health infrastructure and expectations for public health nurses are not clearly defined and therefore pose serious difficulties for conducting disease surveillance. Increased surveillance activities for bioterrorism preparedness add more responsibilities to the work of public health nurses. METHODS: A qualitative study on disease surveillance was conducted with public health officials at regional and local levels, working in a variety of urban and rural settings in one large public health region in Texas. Data analysis was supported by qualitative research software, The Ethnograph. RESULTS: The study found that the nurses working at the local level were extremely dedicated to serving their communities, had formed informal partnerships that are essential for disease surveillance, and effectively used informal communication channels to obtain critical surveillance information. The study revealed that nurses had unmet needs and experienced multiple barriers to conducting disease surveillance. CONCLUSIONS: The study's findings could have implications for public policy and nursing leadership. Defining the structure of the public health system and the scope of public health nurses' responsibilities will serve as the cornerstone for improvement of bioterrorism preparedness. PMID- 16216794 TI - Assessing the readiness and training needs of non-urban physicians in public health emergency and response. AB - Emergency readiness has become a public health priority for United States communities after the 9/11 attacks. Communities that have a less developed public health infrastructure are challenged to organize preparedness and response efforts and to ensure that health care providers are capable of caring for victims of terrorist acts. A survey was used to assess non-urban physicians' prior experience with and self-confidence in treating, and preferred training needs for responding to chemical, biologic, radiologic, nuclear, and explosive (CBRNE) cases. Data were collected through a mailed and Web-based survey. Although the response rate was calculated at 30%, approximately one third of the surveys were not able to be delivered. Most respondents reported never having seen or treated CBRNE-inflicted cases and were not confident in their ability to diagnose or treat CBRNE cases, but many were willing to participate in a state led response plan. Almost half of the individuals had not participated in any related training but expressed interest in receiving training in small group workshops or through CD-ROM. These results provide potential direction for strategic preparedness planning for non-urban health care providers. PMID- 16216795 TI - Development of a state medical surge plan, part I: the procedures, process, and lessons learned or confirmed. AB - In 2003, the Utah Department of Health received funding from the Health Resources and Services Administration to develop a medical surge plan. The plan was designed to increase the number of available hospital beds in the state by 1250 beds, including 125 beds for patients with burns or trauma patients. Interested parties were contacted and a coordinating group composed of Utah Department of Health and University of Utah Health Sciences Center representatives was formed, who were responsible for developing the plan. This article is Part I of a 2-part series that discusses the planning process and identification of a group of stakeholders who served as a planning task force, and concludes with a summary of lessons learned or confirmed during the planning process. Part II will discuss the content of the medical surge plan. PMID- 16216809 TI - Reversibility of hepatocyte nuclear modifications in mice fed on genetically modified soybean. AB - In the literature, the reports on the effects of a genetically modified (GM) diet are scanty and heterogeneous; in particular, no direct evidence has so far been reported that GM food may affect human or animal health. Hepatocytes represent a suitable model for monitoring the effects of a GM diet, the liver potentially being a primary target. In a previous study, we demonstrated that some modifications occur in hepatocyte nuclei of mice fed on GM soybean. In order to elucidate whether such modifications can be reversed, in the present study, 3 months old mice fed on GM soybean since their weaning were submitted to a diet containing wild type soybean, for one month. In parallel, to investigate the influence of GM soybean on adult individuals, mice fed on wild type soybean were changed to a GM diet, for the same time. Using immunoelectron microscopy, we demonstrated that a one-month diet reversion can influence some nuclear features in adult mice, restoring typical characteristics of controls in GM-fed animals, and inducing in control mice modifications similar to those observed in animals fed on GM soybean from weaning. This suggests that the modifications related to GM soybean are potentially reversible, but also that some modifications are inducible in adult organisms in a short time. PMID- 16216810 TI - Rhodiola rosea as antioxidant in red blood cells: ultrastructural and hemolytic behaviour. AB - Rhodiola rosea L. (Crassulaceae) is a plant that lives at high altitude in Europe and Asia, widely used for its high capacity to increase the organism resistance to different stress conditions. Although a few international literature supports these effects, today R. rosea has become a common component of many dietary supplements also in the Western world. The aim of the present study was to investigate the effect of the R. rosea roots aqueous extract on in vitro human erythrocytes exposed to hypochlorous acid (HOCl)-oxidative stress. Several damages occur in human erythrocytes exposed in vitro to HOCl, among these membrane protein and lipid modifications, shifting from the discocyte shape to the echinocyte one, and determining lysis ultimately. Therefore, in the present work, the evaluation of the antioxidant capacity of the Rhodiola extract has been carried out by means of scanning electron microscopy and of hemolytic behaviour on human erythrocytes exposed to HOCl in the presence of increasing doses of the aqueous extract in different experimental environments (co-incubation and subsequent incubations). The results obtained are consistent with a significant protection of the extract in presence of the oxidative agent, but a cautionary note emerges from the analysis of the data related to the cell exposition to the plant extract in the absence of any induced oxidative stress. In fact, the addition to erythrocyte of high doses of R. rosea extract always determines severe alterations of the cell shape. PMID- 16216811 TI - Cytochemical properties of Botryllus schlosseri haemocytes: indications for morpho-functional characterisation. AB - In the present study, we carried out a detailed light microscopy investigation of the cytochemical properties of the haemocytes of the colonial ascidian Botryllus schlosseri, using new cytochemical stains and enzymatic markers, a panel of antibodies and lectins as probes to characterise Botryllus blood cells further. Results indicate that lymphocyte-like cells are circulating undifferentiated cells recognised by anti-CD34 antibody and there are at least two defined haemocyte differentiation pathways: i) phagocytes, represented by hyaline amoebocytes and macrophage-like cells, which share similar staining properties, the same hydrolytic enzyme content as well as the presence of detectable cytochrome-c-oxidase activity, recognition by anti-CD39 and Narcissus pseudonarcissus agglutinin; ii) cytotoxic cell line, represented by granular amoebocytes and morula cells which have vacuoles stained by Ehrlich's stain and Neutral Red; DOPA-containing protein are present inside morula cell vacuoles. Pigment cells and nephrocytes are involved in catabolite storage but their relationships with other cell types are less clear. PMID- 16216812 TI - Spatial distribution of mast cells in chronic venous leg ulcers. AB - Chronic venous leg ulcers (CVUs) show chronic inflammation but different pathological changes occur in different parts of the ulcer. There is a lack of re epithelialisation and defective matrix deposition in the ulcer base but epidermal hyperproliferation and increased matrix deposition in the surrounding skin. The role of mast cells in wound healing, inflammation, fibrosis and epidermal hyperproliferation has been extensively studied but less is known about their role in CVUs. In the present study, we investigated the distribution of mast cells in CVUs with specific consideration of the differences between the ulcer base and the skin surrounding the ulcer. Both histochemical and immunohistological methods were used to detect the mast cell marker tryptase in frozen sections of CVU biopsies. Mast cells were counted in the dermis of normal skin, in the ulcer base and in the skin surrounding the ulcer. Double immunofluorescence staining was used to study the location of mast cells in relation to blood vessels. In normal skin few mast cells were seen in the dermis but none in the epidermis. However in CVUs there was a significant increase in intact and degranulated mast cells in the surrounding skin and ulcer edge (184 per field, p<0.003) of CVUs and a significant reduction in the ulcer base (20.5 per field p<0.05) in comparison to normal skin (61 per field). In CVUs mast cells showed a characteristic location near the epithelial basement membrane whilst mast cell granules and phantom cells (mast cells devoid of granules) were predominantly seen in the epidermis. In the dermis, mast cells were seen associated with blood vessels. The marked increase in mast cells in the surrounding skin of CVUs and depletion of mast cells in the ulcer base could implicate mast cell mediators in the pathological changes in CVUs particularly in the epidermal and vascular changes occurring in the surrounding skin. PMID- 16216813 TI - Productive infection of HUVEC by HHV-8 is associated with changes compatible with angiogenic transformations. AB - Kaposi's Sarcoma (KS) is an angioproliferative disease associated with human herpesvirus 8 (HHV-8) infection. We have characterized the morphologic and phenotypic modifications of HUVEC in a model of productive HHV-8 infection. HHV-8 replication was associated with ultra-structural changes, flattened soma and a loss of marginal folds and intercellular contacts, and morphologic features, spindle cell conversion and cordon-like structures formation. Phenotypic changes observed on cordon-like structures included partial loss and redistribution of CD31/PECAM-1 and VE-cadherin, uPAR up-regulation and de novo expression of CD13/APN. Such changes demonstrate the induction, in HUVEC, of an angiogenic profile. Most of these findings are directly linked to HHV-8-encoded proteins expression, suggesting that HHV-8 itself may participate to the initial steps of the angiogenic transformation in KS. PMID- 16216814 TI - Immunoexpression of CD30 and CD30 ligand in deciduas from spontaneous abortions. AB - In the present study, using immunohistochemistry, we studied the expression of CD30 and CD30-L in 35 deciduas obtained from women following elective abortion during normal physiological gestation and in 60 deciduas obtained from women after spontaneous abortion with or without signs of inflammation. The main difference was noticed in the first trimester of gestation in which was found a decrease in CD30/CD30-L-positive decidual glandular and stromal cells in a greater number of cases of spontaneous abortions with respect to cases of physiological pregnancies (70% vs 50%, p<0.05). In addition, deciduas from spontaneous abortions with inflammation and without inflammation reacted similarly. The reduced expression of CD30 and CD30-L and their cellular pattern detected in the deciduas from spontaneous abortions suggest that the CD30/CD30-L system is crucial for preventing abortions in the first trimester. Furthermore, the distinctive expression of CD30/CD30-L in deciduas from physiological pregnancies may indicate that the CD30/CD30-L system exerts its main role in the first trimester. PMID- 16216816 TI - The distribution of cells containing FMRFamide- and 5-HT-related molecules in the embryonic development of Viviparus ater (Mollusca, Gastropoda). AB - The timing and spatial distribution of cells containing FMRFamide- and 5-HT related molecules in the embryonic development of the mollusc Viviparus ater are examined using immunohistochemistry. FMRFamide-like molecules emerge in the early stage E8 (8% of embryonic development) before the 5-HT immunoreactivity, and they are not only found during nervous system ontogeny. As the parts of the digestive tract differentiated, the pattern of the diffuse gut endocrine cells, present in adults, start to be established (E20-E30), and both open and closed cell types are immunoreactive to anti-FMRFamide antibody. From their appearance (E20), cells with a 5-HT-like phenotype are distributed in the central nervous ganglia and progressively assembled during embryonic development. The early occurrence of both these molecules in V. ater embryos reinforces the growing view that neurotransmitters play a regulatory role in embryogenic processes. In particular, the very early presence of FMRFamide-related factors suggests an involvement of these molecules in the regulation of basic, not only neuronal, cell behaviours, while 5-HT seems to be a more specific neural development signal. PMID- 16216815 TI - Seasonal variations of gonadotropins in the pars distalis male viscacha pituitary. Effect of chronic melatonin treatment. AB - The gonadotropes, LH and FSH cells, were immunohistochemically identified in the pituitary pars distalis of the adult male viscacha (Lagostomus maximus maximus) using specific antibodies against hLHbeta and hFSHbeta with the streptavidin biotin-peroxidase complex. The distribution, size and percentage immunopositive area of these cells were analyzed by image analysis in viscachas captured during the annual reproductive cycle and after the chronic administration of melatonin. The LHbeta and FSHbeta cells showed seasonal changes in the distribution, size and percentage immunopositive area. The LHbeta cells were found widely distributed throughout the pars distalis during the reproductive period, and they were found in the ventro-medial region in the pars distalis during the gonadal regression and gonadal recovery periods. The LHbeta cells reached the largest size and immunopositive area during the reproductive period and the smallest size and immunopositive area during the gonadal regression period. The FSHbeta cells were found in the ventro-medial region during reproductive and gonadal regression periods. The FSHbeta cells were found widely distributed throughout the pars distalis during the gonadal recovery period when they showed the maximum percentage immunopositive area. A decrease in the size of LHbeta and FSHbeta cells was observed after the chronic administration of melatonin. Moreover, it produces a decrease in the immunopositive area occupied by the LHbeta cells but not in the immunopositive area occupied by the FSHbeta cells. Our results show great activity of LHbeta and FSHbeta cells in different moments of the annual reproductive cycle demonstrating that these cells do not secrete in parallel. Moreover, melatonin acts differentially on the activity of the gonadotrope cells. PMID- 16216817 TI - Detection of Epstein Barr virus in formalin-fixed paraffin tissues by fluorescent direct in situ PCR. AB - Specific viral laboratory diagnosis of primary Epstein-Barr Virus (EBV) infection is usually based on antibody-detection assays. However, molecular detection is also considered the reference standard assay for diagnosis of central nervous system infections and of most cases of nasopharyngeal carcinoma (NPC). One-step or nested polymerase chain reaction (PCR) has rapidly replaced immunological assays based on virus-specific Ig antibodies for the laboratory diagnosis of Herpesvirus infections, even if serological methods are considered an additional tool for defining clinical diagnosis. In this article, we will present a rapid, sensitive and robust molecular tool for the viral detection of EBV (EBNA-1) within tissue specimens by making use of in situ PCR (IS-PCR). PMID- 16216818 TI - Law and Psychiatry: mental illness and reasonable accommodations at work: definition of a mental disability under the ADA. PMID- 16216819 TI - Genetic consequence of restricted habitat and population decline in endangered Isoetes sinensis (Isoetaceae). AB - BACKGROUND AND AIMS: Isoetes sinensis (Isoeteaceae) is a critically endangered aquatic quillwort in eastern China. Rapid decline of extant population size and local population extinction have occurred in recent years and have raised great concerns among conservationists. METHODS: Amplified fragment length polymorphisms (AFLPs) were used to investigate the genetic variation and population structure of seven extant populations of the species. KEY RESULTS: Eight primer combinations produced a total of 343 unambiguous bands of which 210 (61.2 %) were polymorphic. Isoetes sinensis exhibited a high level of intra-population genetic diversity (H(E) = 0.118; hs = 0.147; I = 0.192; P = 35.2 %). The genetic variation within each of the populations was not positively correlated with their size, suggesting recent population decline, which is well in accordance with field data of demographic surveys. Moreover, a high degree of genetic differentiation (F(ST) = 0.535; G(ST) = 0.608; theta(B) = 0.607) was detected among populations and no correlation was found between geographical and genetic distance, suggesting that populations were in disequilibrium of migration-drift. Genetic drift played a more important role than gene flow in the current population genetic structure of I. sinensis because migration of I. sinensis is predominantly water-mediated and habitat range was highly influenced by environment changes. CONCLUSIONS: Genetic information obtained in the present study provides useful baseline data for formulating conservation strategies. Conservation management, including both reinforcement for in situ populations and ex situ conservation programmes should be carefully designed to avoid the potential risk of outbreeding depression by admixture of individuals from different regions. However, translocation within the same regional population should be considered as a measure of genetic enhancement to rehabilitate local populations. An ex situ conservation strategy for conserving all extant populations to maximize genomic representation of the species is also recommended. PMID- 16216820 TI - Floral ontogeny in ficinia and isolepis (cyperaceae), with focus on the nature and origin of the gynophore. AB - BACKGROUND AND AIMS: The generic delimitations of Ficinia and Isolepis, sister genera in the Cypereae, are blurred. Typical Ficinia flowers have a lobed gynophore, which envelops the base of the nutlet, whereas in Isolepis the character is considered to be absent. Some former species of Isolepis, lacking the gynophore, were recently included in Ficinia. The floral ontogeny of representative taxa in Ficinia and Isolepis were investigated with the aim of evaluating the origin and nature of the gynophore in the Cypereae. METHODS: The spikelet and floral ontogeny in inflorescences collected in the field was investigated using scanning electron microscopy (SEM) and light microscopy (LM). KEY RESULTS: SEM images of Isolepis setacea and I. antarctica, Ficinia brevifolia, F. minutiflora, F. zeyheri and F. gracilis, and LM sections of F. radiata, show that the gynoecium in Ficinia is elevated above the flower receptacle by the development of a hypogynous stalk. From its apex, a (often three-)lobed cup is formed, which envelopes the basal part of the later nutlet. In developing flowers of I. antarctica, a rudimentary hypogynous stalk appears. In I. setacea, rudiments of a hypogynous stalk can be observed at maturity. In F. radiata and F. zeyheri, intralocular hairs are present in the micropylar zone. At the surface of developing gynoecia in flowers of F. gracilis, star-shaped cuticular structures appear which disappear again at maturity. CONCLUSIONS: The overall floral ontogeny of all species studied occurs following a typical scirpoid pattern, though no perianth primordia are formed. The gynophore in Ficinia originates as a hypogynous stalk, from which the typical gynophore lobes develop. The gynophore is not homologous with the perianth. PMID- 16216821 TI - Self management of oral anticoagulation: randomised trial. AB - OBJECTIVE: To determine the clinical effectiveness of self management compared with routine care in patients on long term oral anticoagulants. DESIGN: Multicentre open randomised controlled trial. SETTING: Midlands region of the UK. PARTICIPANTS: 617 patients aged over 18 and receiving warfarin randomised to intervention (n = 337) and routine care (n = from 2470 invited; 193/337 (57%) completed the 12 month intervention. INTERVENTION: Intervention patients used a point of care device to measure international normalised ratio twice a week and a simple dosing chart to interpret their dose of warfarin. MAIN OUTCOME MEASURE: Percentage of time spent within the therapeutic range of international normalised ratio. RESULTS: No significant differences were found in percentage of time in the therapeutic range between self management and routine care (70% v 68%). Self managed patients with poor control before the study showed an improvement in control that was not seen in the routine care group. Nine patients (2.8/100 patient years) had serious adverse events in the self managed group, compared with seven (2.7/100 patient years) in the routine care arm (chi2(df = 1) = 0.02, P = 0.89). CONCLUSION: With appropriate training, self management is safe and reliable for a sizeable proportion of patients receiving oral anticoagulation treatment. It may improve the time spent the therapeutic range for patients with initially poor control. Trial registration ISRCTN 19313375. PMID- 16216822 TI - Meta-analysis of MTHFR 677C->T polymorphism and coronary heart disease: does totality of evidence support causal role for homocysteine and preventive potential of folate? AB - OBJECTIVES: To investigate the association between the MTHFR 677C-->T polymorphism and coronary heart disease, assessing small study bias and heterogeneity between studies. DATA SOURCES: Medline and Embase citation searches between January 2001 and August 2004; no language restrictions. STUDY SELECTION: Case-control and prospective studies of association between MTHFR 677C-->T variant and myocardial infarction, coronary artery occlusion, or both; 80 studies were included. DATA EXTRACTION: Data on genotype frequency and mean homocysteine concentrations by genotype were extracted. Odds ratios were calculated for TT genotype versus CC genotype. Heterogeneity was explored, with stratification by geographical region of the study samples, and meta-regression by difference in mean serum homocysteine concentrations (CC minus TT genotypes) was carried out. RESULTS: 26,000 cases and 31,183 controls were included. An overall random effects odds ratio of 1.14 (95% confidence intervals 1.05 to 1.24) was found for TT versus CC genotype. There was strong evidence of heterogeneity (P < 0.001, I2 = 38.4%), which largely disappeared after stratification by geographical region. Odds ratios in Europe, Australia, and North America attenuated towards the null, unlike those in the Middle East and Asia. CONCLUSIONS: No strong evidence exists to support an association of the MTHFR 677 C-->T polymorphism and coronary heart disease in Europe, North America, or Australia. Geographical variability may be due to higher folate intake in North America and Europe or to publication bias. The conclusion drawn from previous meta-analyses that folic acid, through lowering homocysteine, has a role in prevention of cardiovascular disease is in some doubt. PMID- 16216823 TI - The role of social isolation in the effects of alcohol on corticosterone and testosterone levels of alcohol-preferring and non-preferring rats. AB - AIMS: Alcohol has been reported to affect the hypothalamic-pituitary-gonadal axis (HPG-axis) and hypothalamic-pituitary-adrenal axis (HPA-axis) as expressed by increased or decreased corticosterone and testosterone levels. Both hormones have also been related to the aetiology of alcohol drinking and the development of alcoholism. Our aim has been to study these interrelations in animal models of alcohol drinking by using social isolation as a model of anxiety. METHODS: The effects of alcohol on serum testosterone and corticosterone concentrations were investigated in alcohol-preferring (AA) and alcohol non-preferring (ANA) rat lines. Animals were tested in mornings and afternoons with 0.75 and 1.5 g alcohol/kg. Half of the animals were kept in single cages, while the control animals were housed in groups of four individuals. RESULTS: The group-caged ANA rats displayed higher control corticosterone levels than the corresponding AA rats during morning sessions (P = 0.007). The AA rats displayed elevated corticosterone levels (AM: P = 0.047) and the ANA rats displayed reduced control corticosterone levels (PM: P = 0.016) in the single cage situation compared with the group-cage situation. Corticosterone concentrations were not affected by low doses and increased (P < 0.05) by high doses of alcohol in all test groups except for isolated AA rats during afternoon sessions. In general, more significant reductions in testosterone levels following alcohol administration were found in the ANA line. In group-caged AA rats, alcohol reduced testosterone levels, while no such effect was observed in isolated AA rats. CONCLUSIONS: We suggest that social isolation, representing stress, may constitute a situation in which the HPA and HPG axes are connected together in promoting alcohol drinking. PMID- 16216824 TI - Pharmacological manipulation of CB1 receptor function alters development of tolerance to alcohol. AB - AIMS: The current study investigated the efficacy of CB1 receptor-targeted drugs on the development and expression of tolerance to alcohol (EtOH). METHODS: An EtOH-inhalation model was used to induce tolerance, as measured by EtOH-induced sedation and hypothermia after a 24 h withdrawal period. Two drug treatment procedures, (i) co-treatment with EtOH and (ii) acute drug administration following chronic EtOH treatment, were used to test the efficacy of CB1 receptor manipulations on EtOH tolerance. RESULTS: The effects of the CB1 receptor agonist CP-55,940 varied depending on paradigm and behavioural measure. Chronic CP-55,940 co-treatment blocked tolerance to EtOH-induced hypothermia but not to the sedative effect (sleep time) in EtOH-exposed mice. However, chronic CP-55,940 administration alone resulted in tolerance to the sedative effect of a challenge dose of EtOH in control mice. Acute CP-55,940 administration after chronic alcoholization blocked the development of tolerance to EtOH-induced sedation compared to the EtOH alone exposed group, but induced tolerance to the hypothermic effects of EtOH in control mice. Chronic blockade of CB1 receptor function by SR141716A resulted in tolerance to both the sedative and hypothermic effects of EtOH in control mice, but had no effect on EtOH-exposed mice. CONCLUSIONS: The data support a role for the endocannabinoid (EC) system in EtOH tolerance/dependence and suggest that drugs targeted against EC system could be therapeutically useful in treating alcohol-related disorders. PMID- 16216825 TI - An optimized strategy for ICAT quantification of membrane proteins. AB - The work presented here focuses on the development of a method adapting isotope labeling of proteins with ICAT to the study of highly hydrophobic proteins. Conditions for the labeling of proteins were first established using two standard soluble proteins and iodoacetamidyl-3,6-dioxaoctanediamine biotin (PEO-iodoacetyl biotin). Results demonstrated the efficiency of the labeling in the presence of high concentrations of both SDS and urea. These conditions were then used to label a highly hydrophobic mitochondrial membrane protein, the adenine nucleotide translocator ANT-1, with PEO-iodoacetyl biotin and then with the cleavable ICAT reagent. The results presented here show that labeling of proteins with cleavable ICAT is possible and may even be improved in strong denaturing buffers containing both SDS at a concentration higher than 0.5% (w/v) and urea. These results open the possibility of applying the ICAT strategy to complex samples containing very hydrophobic proteins solubilized in urea-SDS buffers. The adaptability of the developed method is demonstrated here with preliminary results obtained during the study of membrane-enriched fractions prepared from murine embryonic stem cells. PMID- 16216826 TI - Measuring dust on skin with a small vacuuming sampler--a comparison with other sampling techniques. AB - Airborne skin exposure to allergens and irritants may cause dermatitis. There are few methods for assessing skin exposure to airborne particles. We have modified and tested a vacuuming sampler for removing particles from the skin. The sampler was compared with two other skin and surface exposure sampling techniques. These were based on surrogate skin (a patch sampler-adhesive tape on an optical cover glass) and a tape stripping removal procedure. All three samplers measure the mass of dust on skin. Dust containing starch was deposited onto the skin in a whole-body exposure chamber. Samples were taken from forearms and shoulders and analysed using optical microscopy. With the different sampling techniques small differences in the results were obtained. Agreement between the vacuuming sampler and the tape stripping technique was good. The comparison between patch and tape stripping procedure indicated a slight overestimation for the patch. The three techniques are applicable for assessing skin exposure to particles and for dose effect studies. The vacuuming method will be further developed and applied in workplace studies. The technique allows for dust sampling from large areas of skin. PMID- 16216827 TI - Influence of ghrelin on interdigestive gastrointestinal motility in humans. AB - BACKGROUND: Recent studies in animals have shown that ghrelin stimulates upper gastrointestinal motility through the vagus and enteric nervous system. The aim of the present study therefore was to simultaneously investigate the effect of administration of ghrelin on upper gastrointestinal motility and to elucidate its mode of action by measuring plasma levels of gastrointestinal hormones in humans. MATERIALS AND METHODS: Nine healthy volunteers (four males; aged 22-35 years) underwent combined antroduodenal manometry and proximal stomach barostat study on two separate occasions at least one week apart. Twenty minutes after the occurrence of phase III of the migrating motor complex (MMC), saline or ghrelin 40 mug was administered intravenously over 30 minutes in a double blind, randomised, crossover fashion. Ghrelin, motilin, pancreatic polypeptide, glucagon, and somatostatin were measured by radioimmunoassay in blood samples obtained at 15-30 minute intervals. The influence of ghrelin or saline on MMC phases, hormone levels, and intraballoon volume was compared using paired t test, ANOVA, and chi(2) testing. RESULTS: Spontaneous phase III occurred in all subjects, with a gastric origin in four. Administration of ghrelin induced a premature phase III (12 (3) minutes, p<0.001; gastric origin in nine, p<0.05), compared with saline (95 (13) minutes, gastric origin in two). Intraballoon volumes before infusion were similar (135 (13) v 119 (13) ml; NS) but ghrelin induced a longlasting decrease in intraballoon volume (184 (31) v 126 (21) ml in the first 60 minutes; p<0.05). Administration of ghrelin increased plasma levels of pancreatic polypeptide and ghrelin but motilin, somatostatin, and glucagon levels were not altered. CONCLUSIONS: In humans, administration of ghrelin induces a premature gastric phase III of the MMC, which is not mediated through release of motilin. This is accompanied by prolonged increased tone of the proximal stomach. PMID- 16216828 TI - QUASAR--scoring and ranking of sequence-structure alignments. AB - SUMMARY: Sequence-structure alignments are a common means for protein structure prediction in the fields of fold recognition and homology modeling, and there is a broad variety of programs that provide such alignments based on sequence similarity, secondary structure or contact potentials. Nevertheless, finding the best sequence-structure alignment in a pool of alignments remains a difficult problem. QUASAR (quality of sequence-structure alignments ranking) provides a unifying framework for scoring sequence-structure alignments that aids finding well-performing combinations of well-known and custom-made scoring schemes. Those scoring functions can be benchmarked against widely accepted quality scores like MaxSub, TMScore, Touch and APDB, thus enabling users to test their own alignment scores against 'standard-of-truth' structure-based scores. Furthermore, individual score combinations can be optimized with respect to benchmark sets based on known structural relationships using QUASAR's in-built optimization routines. PMID- 16216829 TI - Syntons, metabolons and interactons: an exact graph-theoretical approach for exploring neighbourhood between genomic and functional data. AB - MOTIVATION: Modern comparative genomics does not restrict to sequence but involves the comparison of metabolic pathways or protein-protein interactions as well. Central in this approach is the concept of neighbourhood between entities (genes, proteins, chemical compounds). Therefore there is a growing need for new methods aiming at merging the connectivity information from different biological sources in order to infer functional coupling. RESULTS: We present a generic approach to merge the information from two or more graphs representing biological data. The method is based on two concepts. The first one, the correspondence multigraph, precisely defines how correspondence is performed between the primary data-graphs. The second one, the common connected components, defines which property of the multigraph is searched for. Although this problem has already been informally stated in the past few years, we give here a formal and general statement together with an exact algorithm to solve it. AVAILABILITY: The algorithm presented in this paper has been implemented in C. Source code is freely available for download at: http://www.inrialpes.fr/helix/people/viari/cccpart. PMID- 16216830 TI - The influence of missing value imputation on detection of differentially expressed genes from microarray data. AB - MOTIVATION: Missing values are problematic for the analysis of microarray data. Imputation methods have been compared in terms of the similarity between imputed and true values in simulation experiments and not of their influence on the final analysis. The focus has been on missing at random, while entries are missing also not at random. RESULTS: We investigate the influence of imputation on the detection of differentially expressed genes from cDNA microarray data. We apply ANOVA for microarrays and SAM and look to the differentially expressed genes that are lost because of imputation. We show that this new measure provides useful information that the traditional root mean squared error cannot capture. We also show that the type of missingness matters: imputing 5% missing not at random has the same effect as imputing 10-30% missing at random. We propose a new method for imputation (LinImp), fitting a simple linear model for each channel separately, and compare it with the widely used KNNimpute method. For 10% missing at random, KNNimpute leads to twice as many lost differentially expressed genes as LinImp. AVAILABILITY: The R package for LinImp is available at http://folk.uio.no/idasch/imp. PMID- 16216831 TI - ATID: a web-oriented database for collection of publicly available alternative translational initiation events. AB - SUMMARY: Alternative translational initiation is an important cellular mechanism contributing to the diversity of protein products and functions. We develop a database that provides a comprehensive collection of alternative translational initiation events. The purpose of this alternative translational initiation database (ATID) is to facilitate the systematic study of alternative translational initiation of genes. The current version of database contains 300 genes from Homo sapiens, Mus musculus and other species. Each of the genes has two or more isoforms due to alternative translational initiation. Resources in ATID, including gene information, alternative products of genes and domain structures of isoforms, are provided through a user-friendly web interface. AVAILABILITY: The ATID database is available for public use at http://bioinfo.au.tsinghua.edu.cn/atie/. PMID- 16216832 TI - CoGenT++: an extensive and extensible data environment for computational genomics. AB - MOTIVATION: CoGenT++ is a data environment for computational research in comparative and functional genomics, designed to address issues of consistency, reproducibility, scalability and accessibility. DESCRIPTION: CoGenT++ facilitates the re-distribution of all fully sequenced and published genomes, storing information about species, gene names and protein sequences. We describe our scalable implementation of ProXSim, a continually updated all-against-all similarity database, which stores pairwise relationships between all genome sequences. Based on these similarities, derived databases are generated for gene fusions--AllFuse, putative orthologs--OFAM, protein families--TRIBES, phylogenetic profiles--ProfUse and phylogenetic trees. Extensions based on the CoGenT++ environment include disease gene prediction, pattern discovery, automated domain detection, genome annotation and ancestral reconstruction. CONCLUSION: CoGenT++ provides a comprehensive environment for computational genomics, accessible primarily for large-scale analyses as well as manual browsing. PMID- 16216833 TI - Variation in survival after diagnosis of breast cancer in Switzerland. AB - BACKGROUND: Survival after diagnosis of cancer is a key criterion for cancer control. Major survival differences between time periods and countries have been reported by the EUROCARE studies. We investigated whether similar differences by period and region existed in Switzerland. METHODS: Survival of 11,376 cases of primary invasive female breast cancer diagnosed between 1988 and 1997 and registered in seven Swiss cancer registries covering a population of 3.5 million was analysed. RESULTS: Comparing the two periods 1988-1992 and 1993-1997, age standardized 5 year relative survival improved globally from 77% to 81%. Furthermore, multivariate analysis adjusting for age, tumour size and nodal involvement identified regional survival differences. Survival was lowest in the rural parts of German-speaking eastern Switzerland and highest in urbanised regions of the Latin- and German-speaking northwestern parts of the country. CONCLUSIONS: This study confirms that survival differences are present even in a small and affluent, but culturally diverse, country like Switzerland, raising the issue of heterogeneity in access to care and quality of treatment. PMID- 16216834 TI - Can exposure to very low levels of asbestos induce pleural mesothelioma? PMID- 16216835 TI - Surfactant dysfunction mutations in children's interstitial lung disease and beyond. PMID- 16216836 TI - Exacerbations: the asthma paradox. PMID- 16216837 TI - Where are the guidelines for the treatment of asthma with panic spectrum symptoms? PMID- 16216839 TI - Systemic versus topical glucocorticoid therapy for acute asthma. PMID- 16216840 TI - OSA brain morphology differences: magnitude of loss approximates age-related effects. PMID- 16216841 TI - Inhaled corticosteroids in wheezy infants. PMID- 16216842 TI - Can the electronic nose really sniff out lung cancer? PMID- 16216843 TI - Identification of cadmium-regulated genes by cDNA-AFLP in the heavy metal accumulator Brassica juncea L. AB - In this study, cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis was employed to identify genes that exhibited a modulated expression following cadmium (Cd) treatment in Brassica juncea grown in hydroponic culture. Plants were treated for 6 h, 24 h, and 6 weeks with 10 microM Cd(NO3)2 and untreated 6 week-old plants were used as controls. Cd content was measured at these four time points. Long exposure to Cd affected root morphology: roots appeared thinner and sent out side roots. Seventy-three transcript-derived fragments were identified as Cd responsive. Fifty-two of them showed significant homology to genes with known or putative function, 10 transcript-derived fragments were homologous to uncharacterized genes, while 11 transcript-derived fragments did not show significant matches. The expression pattern of several of these genes was confirmed by northern blot analysis. Fifty-two genes of known or putative function were transcriptional factors, expression regulators, and stress responding and transport facilitation genes, as well as genes involved in cellular metabolism and organization and the photosynthetic process, suggesting that a multitude of processes are implicated in Cd stress response. The transcription of drought- and abscisic acid-responsive genes observed in this study also suggested that Cd imposes water stress and that abscisic acid may be involved in the Cd plant response. PMID- 16216844 TI - Differential responses of maize MIP genes to salt stress and ABA. AB - Salt stress is known to reduce root hydraulic conductivity and growth. To examine a concomitant regulation of aquaporins, the expression of the maize MIP gene family in response to NaCl was analysed by DNA array hybridization. Plants responded differentially to 100 versus 200 mM NaCl treatments. Leaf water content was reduced rapidly and persistently after the application of 200 mM NaCl in contrast to 100 mM NaCl. Endogenous ABA strongly accumulated in roots after 2 h; it remained at a highly elevated level for 48 h after the addition of 200 mM NaCl, but rapidly declined in plants treated with 100 mM NaCl, indicating an early recovery from water deficit. Interestingly, 2 h after the addition of 100 mM NaCl, when maize regained the osmotic potential allowing water uptake, three highly expressed, specific isoforms ZmPIP1;1, ZmPIP1;5, and ZmPIP2;4 were transiently induced. They were preferentially transcribed in the outer root tissue suggesting a role in cellular water transport. None of the ZmTIP genes was altered. By contrast, after the addition of 200 mM NaCl these responses were missing. Instead, multiple ZmPIP and ZmTIP genes were repressed by 200 mM NaCl after 24 h. After 48 h, deregulations were overridden in both cases indicating homeostasis. ABA (1 muM) exogenously applied to the roots transiently induced ZmPIP2;4 similar to 100 mM NaCl as well as ZmPIP1;2. Thus, the early induction of ZmPIP2;4 by NaCl may be mediated by ABA. Previously, an increase in root hydraulic conductivity had been observed upon ABA application. By contrast, 100 muM ABA led to a complete, possibly non-specific repression of all detected ZmPIP and ZmTIP genes after 24 h. PMID- 16216845 TI - Phosphoenolpyruvate carboxykinase and its potential role in the catabolism of organic acids in the flesh of soft fruit during ripening. AB - Previous studies of grapes and tomatoes have shown that the abundance of phosphoenolpyruvate carboxykinase (PEPCK) increases in their flesh at the start of ripening, and that this coincides with a decrease in its citrate and/or malate content. Thus, PEPCK might function in the catabolism of organic acid anions during the ripening of these fruits. In the present study, the abundance of PEPCK was determined in the flesh of blueberries, raspberries, red currants, and strawberries at different stages of their development. In addition, changes in the amounts of citrate, malate, soluble sugars, isocitrate lyase, NADP-malic enzyme, phosphoenolpyruvate carboxylase, and pyruvate, orthophosphate dikinase in the flesh were determined. PEPCK was not detected in strawberry flesh, in which there was no dissimilation of malate or citrate. In the flesh of the other fruits, the abundance of PEPCK increased during ripening to an amount that was similar to that in grapes and tomatoes. In the flesh of blueberries and red currants, PEPCK was most abundant when there was dissimilation of malate. In the flesh of raspberries, PEPCK was most abundant when there was dissimilation of malate and citrate. These results are consistent with PEPCK playing a role in the dissimilation of citrate and/or malate in the flesh of these fruits during ripening. However, PEPCK was also present in the flesh of blueberries, raspberries, and red currants when there was no dissimilation of malate or citrate, and this raises the possibility that PEPCK might have additional functions. Dissection of blueberries provided evidence that both PEPCK and phosphoenolpyruvate carboxylase were present in the same cells, and possible functions for this are discussed. PMID- 16216846 TI - The environmental plasticity and ecological genomics of the cyanobacterial CO2 concentrating mechanism. AB - Cyanobacteria probably exhibit the widest range of diversity in growth habitats of all photosynthetic organisms. They are found in cold and hot, alkaline and acidic, marine, freshwater, saline, terrestrial, and symbiotic environments. In addition to this, they originated on earth at least 2.5 billion years ago and have evolved through periods of dramatic O2 increases, CO2 declines, and temperature changes. One of the key problems they have faced through evolution and in their current environments is the capture of CO2 and its efficient use by Rubisco in photosynthesis. A central response to this challenge has been the development of a CO2 concentrating mechanism (CCM) that can be adapted to various environmental limitations. There are two primary functional elements of this CCM. Firstly, the containment of Rubisco in carboxysome protein microbodies within the cell (the sites of CO2) elevation), and, secondly, the presence of several inorganic carbon (Ci) transporters that deliver HCO3- intracellularly. Cyanobacteria show both species adaptation and acclimation of this mechanism. Between species, there are differences in the suites of Ci transporters in each genome, the nature of the carboxysome structures and the functional roles of carbonic anhydrases. Within a species, different CCM activities can be induced depending on the Ci availability in the environment. This acclimation is largely based on the induction of multiple Ci transporters with different affinities and specificities for either CO2 or HCO3- as substrates. These features are discussed in relation to our current knowledge of the genomic sequences of a diverse array of cyanobacteria and their ecological environments. PMID- 16216847 TI - Utilization of tomato microarrays for comparative gene expression analysis in the Solanaceae. AB - Transcriptional profiling allows for the assessment and comparison of cross species gene activity and function on a comprehensive scale. The Solanaceae is a large, diverse dicot family, with well-established genetic relationships between major crop species (tomato, potato, pepper, eggplant, and tobacco). Although Arabidopsis thaliana is often the model of choice for anchoring comparative studies, certain biological processes are better examined in other plants. The ripening of fleshy fruits is not tractable in Arabidopsis; however, it has received considerable attention in tomato. As a member of the Solanaceae, tomato provides a well-characterized system to anchor transcriptional profiles of fruit ripening and development in related species. By utilizing different stages of tomato, pepper, and eggplant fruit, the use of tomato microarrays for expression analysis has been demonstrated in closely related heterologous species, and groups of candidate expressed sequence tags, which are useful as orthologous markers, have been identified, as well as genes implicated in fruit ripening and development in the Solanaceae. PMID- 16216848 TI - Cadmium-induced plant stress investigated by scanning electrochemical microscopy. AB - In vivo oxygen evolution above single stomata in Brassica juncea has been used to investigate, for the first time, the effect of Cd-induced stress as imaged by scanning electrochemical microscopy (SECM). SECM images showed a clear stomatal structure-a pore, whose aperture is modulated by two guard cells, serving as the conduit for the oxygen produced. Lower stomatal density and larger stoma size were found in plants treated with 0.2 mM CdCl2 compared with control plants. Either the introduction of Cd caused a slower cell replication in the plane of the epidermis, hence fewer stomata, and/or the number of open stomata was reduced when plants were under Cd-stress. Oxygen evolution above individual stomatal complexes in Cd-treated plants was lower than that from control plants, as determined from the electrochemical current above the middle of each stoma. All guard cells under illumination were swollen, indicating that the stomata were open in both control and treated plants. Thus, decreased oxygen evolution in response to Cd cannot be attributed to simple closing of the stomata, but to a lower photosynthetic yield. SECM provides an excellent tool for monitoring the effects of Cd on photosynthetic activity at the scale of individual stomata. PMID- 16216849 TI - Proteomic analysis of grapevine (Vitis vinifera L.) tissues subjected to herbicide stress. AB - Two-dimensional gel electrophoresis coupled to mass spectrometry analysis was used to examine for the first time the effect of a herbicide (flumioxazin) on a crop species (Vitis vinifera L.) at the proteome level. Examination of 2-D maps derived from chemically stressed tissues revealed the presence of 33 spots displaying a differential expression pattern. The presence of stress responsive proteins in the different plant organs analysed suggests that flumioxazin could act systemically. Among the responsive proteins, some photosynthesis-related proteins, including several fragments of the enzyme Rubisco, were identified. This effect suggests that photosynthesis could be impaired by the herbicide. The induction of several enzymatic antioxidant systems was also observed, probably as a result of an oxidative stress. Moreover, the photorespiration pathway was stimulated, as suggested by the induction of some key enzymes involved in this process. Changes in carbon metabolism-associated proteins presumably reflect altered patterns of carbon flux in response to impaired photosynthesis and an increased need for osmotic adjustment in affected tissues. Finally, plant defences were stimulated as revealed by the induction of a set of proteins belonging to the pathogenesis-related 10 class, suggesting that they could play an essential role in cell defence mechanisms against flumioxazin. PMID- 16216850 TI - Response to bosentan in children with pulmonary hypertension. AB - OBJECTIVE: To describe an early experience of treating 40 children with the dual endothelin receptor antagonist bosentan, which is known to be safe and effective in adults with pulmonary hypertension (PH). DESIGN: In this retrospective, observational study the UK Service for Pulmonary Hypertension for children treated 40 children with bosentan, 20 with idiopathic pulmonary arterial hypertension (IPAH) (mean age 8.03 years, range 1.2-17) and 20 with PH associated with other conditions (congenital heart disease, parenchymal lung or connective tissue disease, or HIV). Their mean age was 8.3 years (range 0.6-16 years). PATIENTS: 39 patients were in World Health Organization (WHO) class III and IV, and all had shown recent deterioration. In IPAH the mean pulmonary vascular resistance (PVR) was 21.7 units.m2 (range 5.6-42.8). In secondary PH the mean PVR was 18 units.m2 (range 4.9-49). No child had a positive response to vasodilator testing with nitric oxide. INTERVENTIONS: Bosentan was given as first line treatment to 25. Nine were given intravenous epoprostenol. Children were treated for a mean of 12.7 months (range 2-24 months). MAIN OUTCOME MEASURES: Response to treatment was judged by WHO functional class, six minute walk test, weight, ECG and echocardiographic findings, and need to add additional treatment. RESULTS: Bosentan was well tolerated. In the IPAH group 19 (95%) stabilised with bosentan treatment but 12 (60%) patients needed combined treatment with epoprostenol. In secondary PH, WHO class, six minute walk test, and weight gain improved significantly. CONCLUSION: Bosentan helped stabilise children with IPAH but intravenous epoprostenol was also needed by 60%. Children with secondary PH improved. PMID- 16216851 TI - Mitral valve reconstruction in the presence of infection. AB - Reconstruction of the mitral valve offers an alternative to replacement in the treatment of active infective endocarditis, with long term benefits for the patient. PMID- 16216852 TI - Antiplatelet therapy and the vascular tree. PMID- 16216854 TI - Cardiac structural and functional abnormalities in end stage renal disease patients with elevated cardiac troponin T. AB - OBJECTIVES: To identify in a prospective observational study the cardiac structural and functional abnormalities and mortality in patients with end stage renal disease (ESRD) with a raised cardiac troponin T (cTnT) concentration. METHODS: 126 renal transplant candidates were studied over a two year period. Clinical, biochemical, echocardiographic, coronary angiographic, and dobutamine stress echocardiographic (DSE) data were examined in comparison with cTnT concentrations dichotomised at cut off concentrations of < 0.04 microg/l and < 0.10 microg/l. RESULTS: Left ventricular (LV) size and filling pressure were significantly raised and LV systolic and diastolic function parameters significantly impaired in patients with raised cTnT, irrespective of the cut off concentration. The proportions of patients with diabetes and on dialysis were higher in both groups with raised cTnT. With a cut off cTnT concentration of 0.04 microg/l but not 0.10 microg/l, significantly more patients had severe coronary artery disease and a positive DSE result. The total ischaemic burden during DSE was similar in cTnT positive and negative patients, irrespective of the cut off concentration used. LV end systolic diameter index and E:Ea ratio were independent predictors of cTnT rises > or = 0.04 microg/l and > or = 0.10 microg/l, respectively. Diabetes was independently associated with cTnT at both cut off concentrations. Mortality was higher in all patients with raised cTnT. CONCLUSIONS: Patients with ESRD with raised cTnT concentrations have increased mortality. Raised concentrations are strongly associated with diabetes, LV dilatation, and impaired LV systolic and diastolic function, but not with severe coronary artery disease. PMID- 16216853 TI - Drug eluting stents: an updated meta-analysis of randomised controlled trials. AB - OBJECTIVE: To confirm the overall benefit of drug eluting stents (DES), to evaluate the effect of different DES, and to assess the global safety of DES compared with bare stents through a meta-analysis of randomised controlled trials. METHODS: Randomised controlled trials comparing sirolimus and derivates or paclitaxel and derivates eluting stents versus bare stents. Binary restenosis and major adverse cardiac events (MACE) were chosen as primary end points. Death, Q wave myocardial infarction (MI), and stent thrombosis up to 12 months' follow up were also analysed. RESULTS: MACE overall occurrence was highly reduced with DES from 19.9% to 10.1% (odds ratio (OR) 0.46, 95% confidence interval (CI) 0.41 to 0.52, p < 0.001). A significant heterogeneity (p < 0.001) was found between subgroups according to the drug: MACE OR was 0.28 (95% CI 0.22 to 0.35) in the sirolimus subgroup and 0.62 (95% CI 0.53 to 0.73) in the paclitaxel subgroup. Restenosis was also highly reduced from 31.7% with bare stents to 10.5% with DES (OR 0.25, 95% CI 0.22 to 0.29, p < 0.001) with a similar heterogeneity between subgroups. Mortality, Q wave MI, and stent thrombosis were not significantly different between DES and control group, whereas Q wave MI and stent thrombosis tended to be more frequent with paclitaxel. CONCLUSION: This meta-analysis confirms the overall benefit of DES on restenosis and MACE with significant heterogeneity between drugs, suggesting higher efficacy of sirolimus eluting stents. Additional data with longer follow up and in high risk populations are needed to clarify issues on stent thrombosis. PMID- 16216855 TI - Historical trends in reported survival rates in patients with hypertrophic cardiomyopathy. AB - OBJECTIVE: To determine the range of survival rates of patients with hypertrophic cardiomyopathy (HCM) by comparing and contrasting the natural history of a cohort of patients seen between 1988 and 2002 with that of other published series. METHODS: 956 adult (> or = 16 years old) patients with HCM (572 men, mean (SD) age 42 (15) years, range 16-88) were evaluated by ECG, Holter, exercise testing, and echocardiography. Patient characteristics and survival data were compared with those in natural history studies from referral and non-referral centres published between 1960 and January 2003. RESULTS: The duration of follow up was 69 (45) months. 120 (12.6%) patients died or underwent cardiac transplantation. Sudden cardiac death (n = 48) was the most common mode of death. The annual rate of sudden death or implantable cardioverter-defibrillator discharge was 1.02 (95% confidence interval (CI) 0.76 to 1.26). Annual rates for heart failure death or transplantation and stroke related death were 0.55% (95% CI 0.37% to 0.78%) and 0.07% (95% CI 0.02% to 0.19%), respectively. When studies published within the last 10 years of the study period were compared with earlier reports, the size of individual study cohorts was larger (309 (240.6) v 136.5 (98.8), p = 0.058) and the proportion with severe functional limitation NYHA class III/IV lower (12.4% v 24.8%, p < 0.0001), and fewer patients underwent septal myotomy-myectomy (5.2% v 18.7%, p < 0.0001). Published sudden death rates over the last 10 years were lower than previously published figures (median 1.0% (range 0.1-1.7) v 2.0% (0 3.5)). CONCLUSION: Published survival rates in HCM cohorts have improved progressively over the past 40 years. In the modern era the prevalence of disease related complications is similar in all reporting centres. PMID- 16216856 TI - Impact on cardiovascular events of increasing high density lipoprotein cholesterol with and without lipid lowering drugs. AB - OBJECTIVE: To estimate the impact on cardiovascular events of changes in high density lipoprotein (HDL) adjusted for changes in total cholesterol. DESIGN: Cohort study based on a record linkage database. SETTING: Community study in Tayside, Scotland, UK. PATIENTS: 18,815 patients were identified for the study between 1989 and 2001. MAIN OUTCOME MEASURES: Cardiovascular events. RESULTS: 5510 patients taking lipid lowering treatment who had not been hospitalised previously for cardiovascular disease had 314 cardiovascular events recorded (9407 person years of follow up). Patients whose HDL rose by > 20% were less likely to have an event (23.5/1000 person years, 95% confidence interval (CI) 17.3 to 29.6) compared with patients whose HDL did not rise (42.6/1000 person years, 95% CI 35.5 to 49.7, adjusted relative risk 0.60, 95% CI 0.44 to 0.83). HDL change and cardiovascular outcome were not significantly associated among patients who had been hospitalised previously for cardiovascular disease or among patients who were not taking lipid lowering drugs. CONCLUSION: In this study a rise in HDL independently predicted reduced cardiovascular risk in patients taking lipid lowering treatment who had not been hospitalised previously for cardiovascular disease. PMID- 16216857 TI - The future of drug eluting stents. AB - In-stent restenosis (ISR) is the major drawback of percutaneous coronary interventions, occurring in 10-40% of patients. Drug eluting stents (DES) are successful in a large majority of patients in preventing restenosis for the first year after implantation. Recently, new stents have emerged that are loaded with anti-inflammatory, antimigratory, antiproliferative, or pro-healing drugs. These drugs are supposed to inhibit inflammation and neointimal growth and subsequently ISR. The future of DES lies in the development of better stents with new stent designs, better polymers including biological polymers and biological biodissolvable stent coatings, and new, better drugs. PMID- 16216858 TI - Relation between aortic stiffness and coronary flow reserve in patients with coronary artery disease. AB - OBJECTIVES: To investigate the relation between aortic stiffness and coronary flow reserve (CFR) in patients with coronary artery disease (CAD). DESIGN: Observational study. SETTING: Coronary care unit of a primary care hospital. PATIENTS: 192 consecutive patients who underwent coronary angiography. MAIN OUTCOME MEASURE: Brachial-ankle pulse wave velocity (ba-PWV), CFR, and severity of CAD. RESULTS: According to the angiographic findings, patients were divided into four subgroups: patients without significant stenosis (normal coronary artery (NCA) group, n = 28) and those with one vessel disease (1VD group, n = 92), two vessel disease (2VD group, n = 50), or three vessel disease (3VD group, n = 22). ba-PWV increased with the number of diseased vessels and was significantly correlated with the number of diseased vessels (NCA group v 1VD group v 2VD group v 3VD group: 1481 (252) v 1505 (278) v 1577 (266) v 1727 (347) cm/s, p < 0.001). CFR had a significant negative correlation with ba-PWV (r = 0.45, p < 0.0001). The diastolic to systolic velocity ratio obtained in 45 patients also was significantly correlated with ba-PWV (r = -0.35, p < 0.05). Multiple regression analysis showed that ba-PWV was an independent determinant of CFR (p < 0.01). CONCLUSIONS: Coronary flow is altered with aortic stiffening in patients with CAD. These results suggest one possible mechanism for recent reports that aortic stiffness is a key cardiovascular risk factor. PMID- 16216859 TI - Does a well developed collateral circulation predispose to restenosis after percutaneous coronary intervention? An intravascular ultrasound study. AB - OBJECTIVE: To evaluate whether a well developed collateral circulation predisposes to restenosis after percutaneous coronary intervention (PCI). DESIGN: Prospective observational study. PATIENTS AND SETTING: 58 patients undergoing elective single vessel PCI in a tertiary referral interventional cardiac unit in the UK. METHODS: Collateral flow index (CFI) was calculated as (Pw-Pv)/(Pa-Pv), where Pa, Pw, and Pv are aortic, coronary wedge, and right atrial pressures during maximum hyperaemia. Collateral supply was considered poor (CFI < 0.25) or good (CFI > or = 0.25). MAIN OUTCOME MEASURES: In-stent restenosis six months after PCI, classified as neointimal volume > or = 25% stent volume on intravascular ultrasound (IVUS), or minimum lumen area < or = 50% stent area on IVUS, or minimum lumen diameter < or = 50% reference vessel diameter on quantitative coronary angiography. RESULTS: Patients with good collaterals had more severe coronary stenoses at baseline (90 (11)% v 75 (16)%, p < 0.001). Restenosis rates were similar in poor and good collateral groups (35% v 43%, p = 0.76 for diameter restenosis, 27% v 45%, p = 0.34 for area restenosis, and 23% v 24%, p = 0.84 for volumetric restenosis). CFI was not correlated with diameter, area, or volumetric restenosis (r2 < 0.1 for each). By multivariate analysis, stent diameter, stent length, > 10% residual stenosis, and smoking history were predictive of restenosis. CONCLUSION: A well developed collateral circulation does not predict an increased risk of restenosis after PCI. PMID- 16216860 TI - The changing face of infective endocarditis. AB - Infective endocarditis (IE) is an evolving disease with a persistently high mortality and morbidity, even in the modern era of advanced diagnostic imaging, improved antimicrobial chemotherapy, and potentially curative surgery. Despite these improvements in health care, the incidence of the disease has remained unchanged over the past two decades and may even be increasing. Chronic rheumatic heart disease is now an uncommon antecedent, whereas degenerative valve disease of the elderly, mitral valve prolapse, intravenous drug misuse, preceding valve replacement, and vascular instrumentation have become increasingly common, coinciding with an increase in staphylococcal infections and those caused by fastidious organisms. The current understanding of this difficult condition is reviewed and recent developments in medical and surgical management are updated. PMID- 16216861 TI - Left ventricular adaptation after atrial septal defect closure assessed by increased concentrations of N-terminal pro-brain natriuretic peptide and cardiac magnetic resonance imaging in adult patients. AB - OBJECTIVE: To determine in an observational study whether N-terminal pro-brain natriuretic peptide (NT-proBNP) is raised in patients with an atrial septal defect (ASD) and whether concentrations change after interventional closure. METHODS: 12 patients (6 men, mean (SD) age 44.4 (18.6) years) with a moderate sized ASD type II (23.3 (4.5) mm, pulmonary to systemic flow ratio 2.1 (0.68)) were investigated. In all patients a magnetic resonance imaging (MRI) study was performed and NT-proBNP was assessed at baseline and early (9 (13) days) and late (138 (64) days) after intervention. RESULTS: Concentrations of NT-proBNP were found to be within the normal range at baseline (median 87 pg/ml, interquartile range 65-181 pg/ml) but increased early after the interventional closure (315 pg/ml, 133-384 pg/ml, p = 0.005 versus baseline). The increase of NT-proBNP was associated with an increase in left ventricular dimensions as assessed by MRI (left ventricular end diastolic volume 104 (27) ml to 118 (27) ml, p = 0.003). Late after ASD closure NT-proBNP returned to baseline concentrations (102 pg/ml, 82-188 pg/ml, p = 0.004 versus early follow up). CONCLUSION: These findings suggest the presence of transitory haemodynamic stress during adaptation of the left ventricle after ASD closure, which may contribute to the understanding of the pathological mechanism of acute heart failure and delayed improvement of exercise capacity after ASD closure. PMID- 16216862 TI - Socioeconomic status and ischaemic heart disease mortality in 10 western European populations during the 1990s. AB - OBJECTIVE: To assess the association between socioeconomic status and ischaemic heart disease (IHD) mortality in 10 western European populations during the 1990s. DESIGN: Longitudinal study. SETTING: 10 European populations (95,009,822 person years). METHODS: Longitudinal data on IHD mortality by educational level were obtained from registries in Finland, Norway, Denmark, England/Wales, Belgium, Switzerland, Austria, Turin (Italy), Barcelona (Spain), and Madrid (Spain). Age standardised rates and rate ratios (RRs) of IHD mortality by educational level were calculated by using Poisson regression. RESULTS: IHD mortality was higher in those with a lower socioeconomic status than in those with a higher socioeconomic status among men aged 30-59 (RR 1.55, 95% confidence interval (CI) 1.51 to 1.60) and 60 years and over (RR 1.22, 95% CI 1.21 to 1.24), and among women aged 30-59 (RR 2.13, 95% CI 1.98 to 2.29) and 60 years and over (RR 1.36, 95% CI 1.33 to 1.38). Socioeconomic disparities in IHD mortality were larger in the Scandinavian countries and England/Wales, of moderate size in Belgium, Switzerland, and Austria, and smaller in southern European populations among men and younger women (p < 0.0001). For elderly women the north-south gradient was smaller and there was less variation between populations. No socioeconomic disparities in IHD mortality existed among elderly men in southern Europe. CONCLUSIONS: Socioeconomic disparities in IHD mortality were larger in northern than in southern European populations during the 1990s. This partly reflects the pattern of socioeconomic disparities in cardiovascular risk factors in Europe. Population wide strategies to reduce risk factor prevalence combined with interventions targeted at the lower socioeconomic groups can contribute to reduce IHD mortality in Europe. PMID- 16216863 TI - Anaemia is associated with higher mortality among patients with heart failure with preserved systolic function. AB - OBJECTIVE: To investigate the prevalence of anaemia and its influence on mortality among hospitalised patients with congestive heart failure (CHF) with preserved left ventricular systolic function (LVSF). METHOD AND RESULTS: 210 patients with preserved LVSF admitted to the cardiology department of a tertiary hospital for CHF between 1 January 2000 and 31 December 2002 were analysed. Anaemic patients, who constituted 46% of the whole group, were older (75 v 72 years, p = 0.036); were in hospital longer (mean (SD) 13 v 11 days, p = 0.007); had a higher prevalence of ischaemic heart disease (54% v 35%, p = 0.009), left bundle branch block (12% v 4%, p = 0.018), and kidney failure (56% v 34%, p = 0.003); and had faster erythrocyte sedimentation rates (mean (SD) 50 v 26 mm in the first hour, p < 0.001), a tendency to lower serum cholesterol concentration (mean (SD) 4.65 v 5.22 mmol/l, p = 0.073), and smaller body mass index (mean (SD) 27 v 29 kg/m2, p = 0.126) than their non-anaemic counterparts. Kaplan-Meier analysis showed the anaemic group to have significantly poorer survival (p = 0.0001), with a one year survival rate of 72.2% versus 90.5% in the non-anaemic group. Multivariate analysis showed anaemia to be the most powerful independent predictor of mortality, increasing the risk of death by a factor of 2.7 (p = 0.007). CONCLUSION: Anaemia is a very prevalent condition in hospitalised patients with CHF with preserved LVSF and is independently associated with higher mortality. Appropriately designed randomised studies are needed to determine whether the prevention or treatment of anaemia can improve survival of these patients. PMID- 16216864 TI - Effect of statins on the mortality of patients with ischaemic heart disease: population based cohort study with nested case-control analysis. AB - OBJECTIVE: To measure the effect of statins on mortality for community based patients with ischaemic heart disease and determine whether the likely benefits are similar for women, the elderly, and patients with diabetes. DESIGN: Open prospective cohort study with nested case-control analysis. SETTING: 1.18 million patients registered with 89 practices spread across 23 strategic health authority areas within the UK. All practices had a minimum of eight years of longitudinal data and were contributing to the UK QRESEARCH database. SUBJECTS: All patients with a first diagnosis of ischaemic heart disease between January 1996 and December 2003. OUTCOMES: Adjusted hazard ratio with 95% confidence intervals (CIs) for all cause mortality (cohort analysis) and odds ratio (OR) with 95% CI (case-control analysis) for current use of statins. Adjustments were made for current use of aspirin, beta blockers, and angiotensin converting enzyme inhibitors, co-morbidity (myocardial infarction, diabetes, hypertension, congestive cardiac failure), smoking, body mass index, and quintile of deprivation. RESULTS: 13,029 patients had a first diagnosis of ischaemic heart disease in the study period giving an incidence rate of 3.38/1000 person years. 2266 patients with ischaemic heart disease died during the 43,460 person years of observation giving an overall mortality rate of 52.1/1000 person years (95% CI 50.0 to 54.3). In the case-control analysis, patients taking statins had a 39% lower risk of death than did patients not taking statins (adjusted OR 0.61, 95% CI 0.52 to 0.72) after use of other medication, co-morbidity, smoking, body mass index, and deprivation were taken into account. The benefits found in this study compared favourably with those found in the randomised controlled trials, although the current study population is at higher overall risk. The benefits extend to women, patients with diabetes, and the elderly and can be seen within two years of treatment. Longer duration of usage was associated with lower OR for risk of death with a 19% reduction in risk of death with each additional year of treatment (adjusted OR 0.81, 95% CI 0.77 to 0.86 per year). Mortality was similarly reduced among patients prescribed atorvastatin (adjusted OR 0.62, 95% CI 0.48 to 0.79) and simvastatin (adjusted OR 0.62, 95% CI 0.50 to 0.76). CONCLUSIONS: The benefits of statins found in randomised controlled trials extend to unselected community based patients. The benefits can be seen within the first two years of treatment and continue to accrue over time. Since patients in the community are likely to be at higher risk than those in trials, the potential benefits from statins are likely to be greater than expected. PMID- 16216865 TI - B-type natriuretic peptide identifies silent myocardial ischaemia in stroke survivors. AB - OBJECTIVE: To test the hypothesis that B-type natriuretic peptide (BNP) predicts reversible myocardial ischaemia in stroke survivors who do not have chest pain or previous myocardial infarction. METHODS: 56 stroke survivors (mean (SE) age 68 (8) years) underwent tetrofosmin myocardial perfusion scanning with dipyridamole as the stressor. The degree of ischaemia was assessed by a scoring system (out of 64) by an experienced observer blinded to the results of BNP. RESULTS: In the whole cohort, BNP was significantly correlated with the degree of myocardial ischaemia on stress scanning (Spearman's r = -0.475, p < 0.001). BNP also correlated with the degree of reversible ischaemia (stress score - rest score; Spearman's r = 0.28, two tailed p = 0.049). In the cohort who did not have left ventricular systolic dysfunction (n = 44), BNP remained higher in patients with relevant myocardial ischaemia (mean (SE) BNP 20.9 pg/ml, 95% confidence interval (CI) 15.2 to 26.5 v 12.2 pg/ml, 95% CI 5.95 to 18.5; p = 0.046); 33 of the 44 patients had no chest pain or history of myocardial infarction. The relation between resting BNP and both inducible ischaemia and dipyridamole stress score remained significant (Spearman's r = 0.37 and -0.38, respectively). CONCLUSIONS: BNP correlates with the degree of reversible myocardial ischaemia in patients who do not have chest pain or a history of myocardial infarction or evidence of left ventricular systolic dysfunction. Stroke survivors with a high BNP deserve further investigations to rule out significant reversible myocardial ischaemia, in order to reduce their risk of cardiac death. PMID- 16216866 TI - Natriuretic peptides as a prognostic marker and therapeutic target in heart failure. AB - Natriuretic peptides may have an increasing role in assisting clinicians to target treatment in patients with chronic heart failure. PMID- 16216867 TI - Detection of oligomerization and conformational changes in the Na+/H+ antiporter from Helicobacter pylori by fluorescence resonance energy transfer. AB - Oligomerization and conformational changes in the Na+/H+ antiporter from Helicobacter pylori (HPNhaA) were studied by means of fluorescence resonance energy transfer (FRET) analysis. Na+/H+ antiporter-deficient Escherichia coli cells expressing C-terminal fusions of HPNhaA to green fluorescent protein (GFP) variants exhibited wild-type levels of antiporter activity in their everted membrane vesicles. Vesicles containing both HPNhaA-CFP and HPNhaA-YFP or HPNhaA Venus exhibited FRET from CFP (donor) to YFP or Venus (acceptor), suggesting that HPNhaA forms an oligomer. Co-precipitation of HPNhaA tagged by Venus and FLAG sequences confirmed oligomerization. FRET decreased extensively after treatment of the vesicles with proteinase K, which released GFP variants from the fusion proteins. FRET was not observed by merely mixing vesicles expressing the donor or acceptor fusion alone. Fluorescence of Venus is less sensitive to anions and stronger than that of anion-sensitive YFP. Using HPNhaA-Venus as the acceptor, Li+ was found to cause a significant decrease in FRET regardless of the presence or absence of DeltapH across the membranes, whereas Na+ caused a much weaker effect. This Li+ effect was minimal in vesicles prepared from cells expressing HPNhaA containing an Asp141 to Asn mutation, which results in defective Li+/H+ antiporter activity, possibly Li+ binding. These results demonstrate that monomer interactions within the HPNhaA oligomer are weakened possibly by Li+ binding. Dynamic interactions between HPNhaA monomers were detectable in membranes by FRET analysis, thus providing a new approach to study dynamic conformational changes in NhaA during antiport activity. PMID- 16216868 TI - Subplasmalemmal mitochondria modulate the activity of plasma membrane Ca2+ ATPases. AB - Mitochondria are dynamic organelles that modulate cellular Ca2+ signals by interacting with Ca2+ transporters on the plasma membrane or the endoplasmic reticulum (ER). To study how mitochondria dynamics affects cell Ca2+ homeostasis, we overexpressed two mitochondrial fission proteins, hFis1 and Drp1, and measured Ca2+ changes within the cytosol and the ER in HeLa cells. Both proteins fragmented mitochondria, decreased their total volume by 25-40%, and reduced the fraction of subplasmalemmal mitochondria by 4-fold. The cytosolic Ca2+ signals elicited by histamine were unaltered in cells lacking subplasmalemmal mitochondria as long as Ca2+ was present in the medium, but the signals were significantly blunted when Ca2+ was removed. Upon Ca2+ withdrawal, the free ER Ca2+ concentration decreased rapidly, and hFis1 cells were unable to respond to repetitive histamine stimulations. The loss of stored Ca2+ was due to an increased activity of plasma membrane Ca2+-ATPase (PMCA) pumps and was associated with an increased influx of Ca2+ and Mn2+ across store-operated Ca2+ channels. The increased Ca2+ influx compensated for the loss of stored Ca2+, and brief Ca2+ additions between successive agonist stimulations fully corrected subsequent histamine responses. We propose that the lack of subplasmalemmal mitochondria disrupts the transfer of Ca2+ from plasma membrane channels to the ER and that the resulting increase in subplasmalemmal [Ca2+] up-regulates the activity of PMCA. The increased Ca2+ extrusion promotes ER depletion and the subsequent activation of store-operated Ca2+ channels. Cells thus adapt to the lack of subplasmalemmal mitochondria by relying on external rather than on internal Ca2+ for signaling. PMID- 16216869 TI - Peroxisome proliferator-activated receptor gamma suppresses proximal alpha1(I) collagen promoter via inhibition of p300-facilitated NF-I binding to DNA in hepatic stellate cells. AB - Depletion of peroxisome proliferator-activated receptor gamma (PPARgamma) represents one of the key molecular changes that underlie transdifferentiation (activation) of hepatic stellate cells in the genesis of liver fibrosis (Miyahara, T., Schrum, L., Rippe, R., Xiong, S., Yee, H. F., Jr., Motomura, K., Anania, F. A., Willson, T. M., and Tsukamoto, H. (2000) J. Biol. Chem. 275, 35715 35722; Hazra, S., Xiong, S., Wang, J., Rippe, R. A., Krishna, V., Chatterjee, K., and Tsukamoto, H. (2004) J. Biol. Chem. 279, 11392-11401). In support of this notion, ectopic expression of PPARgamma suppresses hepatic stellate cells activation markers, most notably expression of alpha1(I) procollagen. However, the mechanisms underlying this antifibrotic effect are largely unknown. The present study utilized deletion-reporter gene constructs of proximal 2.2-kb alpha1(I) procollagen promoter to demonstrate that a region proximal to -133 bp is where PPARgamma exerts its inhibitory effect. Within this region, two DNase footprints with Sp1 and reverse CCAAT box sites exist. NF-I, but not CCAAT DNA binding factor/NF-Y, binds to the proximal CCAAT box in hepatic stellate cells. A mutation of this site almost completely abrogates the promoter activity. NF-I mildly but independently stimulates the promoter activity and synergistically promotes Sp1-induced activity. PPARgamma inhibits NF-I binding to the most proximal footprint (-97/-85 bp) and inhibits its transactivity. The former effect is mediated by the ability of PPARgamma to inhibit p300-facilitated NF-I binding to DNA as demonstrated by chromatin immunoprecipitation assay. PMID- 16216870 TI - Structural basis for activation of the thiamin diphosphate-dependent enzyme oxalyl-CoA decarboxylase by adenosine diphosphate. AB - Oxalyl-coenzyme A decarboxylase is a thiamin diphosphate-dependent enzyme that plays an important role in the catabolism of the highly toxic compound oxalate. We have determined the crystal structure of the enzyme from Oxalobacter formigenes from a hemihedrally twinned crystal to 1.73 A resolution and characterized the steady-state kinetic behavior of the decarboxylase. The monomer of the tetrameric enzyme consists of three alpha/beta-type domains, commonly seen in this class of enzymes, and the thiamin diphosphate-binding site is located at the expected subunit-subunit interface between two of the domains with the cofactor bound in the conserved V-conformation. Although oxalyl-CoA decarboxylase is structurally homologous to acetohydroxyacid synthase, a molecule of ADP is bound in a region that is cognate to the FAD-binding site observed in acetohydroxyacid synthase and presumably fulfils a similar role in stabilizing the protein structure. This difference between the two enzymes may have physiological importance since oxalyl-CoA decarboxylation is an essential step in ATP generation in O. formigenes, and the decarboxylase activity is stimulated by exogenous ADP. Despite the significant degree of structural conservation between the two homologous enzymes and the similarity in catalytic mechanism to other thiamin diphosphate-dependent enzymes, the active site residues of oxalyl-CoA decarboxylase are unique. A suggestion for the reaction mechanism of the enzyme is presented. PMID- 16216872 TI - Multifunctional roles of the conserved Arg residues in the second region of homology of p97/valosin-containing protein. AB - The 97-kDa molecular chaperone valosin-containing protein (VCP) belongs to a highly conserved AAA family and forms a hexameric structure that is essential for its biological functions. The AAA domain contains highly conserved motifs, the Walker A, Walker B, and the second region of homology (SRH). Although Walker A and B motifs mediate ATP binding and hydrolysis, respectively, the function of the SRH in VCP is not clear. We examined the significance of the SRH in VCP, especially the conserved Arg(359) and Arg(362) in the first AAA domain, D1 and Arg(635) and Arg(638) in the second AAA domain, D2. We show that Arg(359) and Arg(362) in D1 are critical for maintaining the hexameric structure and the ability to bind the polyubiquitin chains. Although the rest of the tested SRH mutants retain the hexameric structure, all of them exhibit severely reduced ATPase activity. Tryptophan fluorescence analysis showed that all of the tested mutants can bind to ATP or ADP. Thus, the reduced ATPase activity likely results from the hampered communications among protomers during hydrolysis. Moreover, when the ATPase-defective mutant R635A or R638A is mixed with the Walker A mutant of D2, the ATPase activity is partially restored, suggesting that Arg(635) and Arg(638) can stimulate the ATPase activity of the neighboring protomer. Interestingly, mutation of Arg(359) and Arg(362) uncouples the inhibitory effect of p47, a VCP co-factor, on the ATPase activity of VCP. Therefore, the Arg residues allow D1 to take on a specific conformation that is required for substrate binding and co-factor communications. Taken together, these results demonstrate that the conserved Arg residues in the SRH of both D1 and D2 play critical roles in communicating the conformational changes required for ATP hydrolysis, and SRH in D1 also contributes to substrate binding and co-factor communications. PMID- 16216871 TI - The docosatriene protectin D1 is produced by TH2 skewing and promotes human T cell apoptosis via lipid raft clustering. AB - Docosahexaenoic acid, a major omega-3 fatty acid in human brain, synapses, retina, and other neural tissues, displays beneficial actions in neuronal development, cancer, and inflammatory diseases by mechanisms that remain to be elucidated. In this study we found, using lipid mediator informatics employing liquid chromatography-tandem mass spectrometry, that (10,17S) docosatriene/neuroprotectin D1, now termed protectin D1 (PD1), is generated from docosahexaenoic acid by T helper type 2-skewed peripheral blood mononuclear cells in a lipoxygenase-dependent manner. PD1 blocked T cell migration in vivo, inhibited tumor necrosis factor alpha and interferon-gamma secretion, and promoted apoptosis mediated by raft clustering. These results demonstrated novel anti-inflammatory roles for PD1 in regulating events associated with inflammation and resolution. PMID- 16216874 TI - Lecithin:retinol acyltransferase is responsible for amidation of retinylamine, a potent inhibitor of the retinoid cycle. AB - Lecithin:retinol acyltransferase (LRAT) catalyzes the transfer of an acyl group from the sn-1 position of phosphatidylcholine to all-trans-retinol (vitamin A) and plays an essential role in the regeneration of visual chromophore as well as in the metabolism of vitamin A. Here we demonstrate that retinylamine (Ret-NH2), a potent and selective inhibitor of 11-cis-retinal biosynthesis (Golczak, M., Kuksa, V., Maeda, T., Moise, A. R., and Palczewski, K. (2005) Proc. Natl. Acad. Sci. U. S. A. 102, 8162-8167), is a substrate for LRAT. LRAT catalyzes the transfer of the acyl group onto Ret-NH2 leading to the formation of N retinylpalmitamide, N-retinylstearamide, and N-retinylmyristamide with a ratio of 15:6:2, respectively. The presence of N-retinylamides was detected in vivo in mice supplemented with Ret-NH2. N-Retinylamides are thus the main metabolites of Ret-NH2 in the liver and the eye and can be mobilized by hydrolysis/deamidation back to Ret-NH2. Using two-photon microscopy and the intrinsic fluorescence of N retinylamides, we showed that newly formed amides colocalize with the retinyl ester storage particles (retinosomes) in the retinal pigment epithelium. These observations provide new information concerning the substrate specificity of LRAT and explain the prolonged effect of Ret-NH2 on the rate of 11-cis-retinal recovery in vivo. PMID- 16216873 TI - S100A8 and S100A9 in human arterial wall. Implications for atherogenesis. AB - Atherogenesis is a complex process involving inflammation. S100A8 and S100A9, the Ca2+-binding neutrophil cytosolic proteins, are associated with innate immunity and regulate processes leading to leukocyte adhesion and transmigration. In neutrophils and monocytes the S100A8-S100A9 complex regulates phosphorylation, NADPH-oxidase activity, and fatty acid transport. The proteins have anti microbial properties, and S100A8 may play a role in oxidant defense in inflammation. Murine S100A8 is regulated by inflammatory mediators and recruits macrophages with a proatherogenic phenotype. S100A9 but not S100A8 was found in macrophages in ApoE-/- murine atherosclerotic lesions, whereas both proteins are expressed in human giant cell arteritis. Here we demonstrate S100A8 and S100A9 protein and mRNA in macrophages, foam cells, and neovessels in human atheroma. Monomeric and complexed forms were detected in plaque extracts. S100A9 was strongly expressed in calcifying areas and the surrounding extracellular matrix. Vascular matrix vesicles contain high levels of Ca2+-binding proteins and phospholipids that regulate calcification. Matrix vesicles characterized by electron microscopy, x-ray microanalysis, nucleoside triphosphate pyrophosphohydrolase assay and cholesterol/phospholipid analysis contained predominantly S100A9. We propose that S100A9 associated with lipid structures in matrix vesicles may influence phospholipid-Ca2+ binding properties to promote dystrophic calcification. S100A8 and S100A9 were more sensitive to hypochlorite oxidation than albumin or low density lipoprotein and immunoaffinity confirmed S100A8-S100A9 complexes; some were resistant to reduction, suggesting that hypochlorite may contribute to protein cross-linking. S100A8 and S100A9 in atherosclerotic plaque and calcifying matrix vesicles may significantly influence redox- and Ca2+-dependent processes during atherogenesis and its chronic complications, particularly dystrophic calcification. PMID- 16216875 TI - Identification of six novel allosteric effectors of Arabidopsis thaliana aspartate kinase-homoserine dehydrogenase isoforms. Physiological context sets the specificity. AB - The Arabidopsis genome contains two genes predicted to code for bifunctional aspartate kinase-homoserine dehydrogenase enzymes (isoforms I and II). These two activities catalyze the first and the third steps toward the synthesis of the essential amino acids threonine, isoleucine, and methionine. We first characterized the kinetic and regulatory properties of the recombinant enzymes, showing that they mainly differ with respect to the inhibition of the homoserine dehydrogenase activity by threonine. A systematic search for other allosteric effectors allowed us to identify an additional inhibitor (leucine) and 5 activators (alanine, cysteine, isoleucine, serine, and valine) equally efficient on aspartate kinase I activity (4-fold activation). The six effectors of aspartate kinase I were all activators of aspartate kinase II activity (13-fold activation) and displayed a similar specificity for the enzyme. No synergy between different effectors could be observed. The activation, which resulted from a decrease in the Km values for the substrates, was detected using low substrates concentrations. Amino acid quantification revealed that alanine and threonine were much more abundant than the other effectors in Arabidopsis leaf chloroplasts. In vitro kinetics in the presence of physiological concentrations of the seven allosteric effectors confirmed that aspartate kinase I and II activities were highly sensitive to changes in alanine and threonine concentrations. Thus, physiological context rather than enzyme structure sets the specificity of the allosteric control. Stimulation by alanine may play the role of a feed forward activation of the aspartate-derived amino acid pathway in plant. PMID- 16216876 TI - Low energy pathways and non-native interactions: the influence of artificial disulfide bridges on the mechanism of folding. AB - Four versions of a beta-sheet protein (CD2.d1) have been made, each with a single artificial disulfide bond inserted into hairpin structures. Folding kinetics of reduced and oxidized forms shows bridge position strongly influences its effect on the folding reaction. Bridging residues 58 and 62 does not affect the rapidly formed intermediate (I) or rate-limiting transition (t) state, whereas bridging 33 and 38, or 31 and 41, lowers the t-state energy, with the latter having the stronger influence. Bridging residues 79 and 90 stabilizes both I- and t-states. To assess additivity in the energetic effects of these bridges, four double bridge variants have also been made. All show precise additivity of overall stability, with two showing additivity when ground states and the rate-limiting t state are assessed, i.e. no measurable change in the folding mechanism occurs. However, combining 31-41 and 79-90 bridges produces a molecule that folds through a different pathway, with a much more stable intermediate than expected and a much higher t-state barrier. This is explained by the artificial introduction of stabilizing, non-native contacts in the I-state. More surprisingly, for another double-bridge version (58-62 and 79-90) both I- and t-states are less stable than expected, showing that conformational constraints introduced by the two bridges prevent formation of non-native contacts that would otherwise stabilize the I- and t-states, thereby lowering the energy of the folding landscape in the wild type (unbridged) molecule. We conclude that the lowest energy path for folding has I- and t-state structures that are stabilized by non-native interactions. PMID- 16216877 TI - Subunit a of the yeast V-ATPase participates in binding of bafilomycin. AB - Bafilomycin and concanamycin are potent and highly specific inhibitors of the vacuolar (H(+))-ATPases (V-ATPases), typically inhibiting at nanomolar concentrations. Previous studies have shown that subunit c of the integral V(0) domain participates in bafilomycin binding, and that this site resembles the oligomycin binding site of the F-ATPase (Bowman, B. J., and Bowman, E. J. (2002) J. Biol. Chem. 277, 3965-3972). Because mutations in F-ATPase subunit a also confer resistance to oligomycin, we investigated whether the a subunit of the V ATPase might participate in binding bafilomycin. Twenty-eight subunit a mutations were constructed just N-terminal to the critical Arg(735) residue in transmembrane 7 required for proton transport, a region similar to that shown to participate in oligomycin binding by the F-ATPase. The mutants appeared to assemble normally and all but two showed normal growth at pH 7.5, whereas all but three had at least 25% of wild-type levels of proton transport and ATPase activity. Of the functional mutants, three displayed K(i) values for bafilomycin significantly different from wild-type (0.22 +/- 0.03 nm). These included E721K (K(i) 0.38 +/- 0.03 nm), L724A (0.40 +/- 0.02 nm), and N725F (0.54 +/- 0.06 nm). Only the N725F mutation displayed a K(i) for concanamycin (0.84 +/- 0.04 nm) that was slightly higher than wild-type (0.60 +/- 0.07 nm). These results suggest that subunit a of V-ATPase participates along with subunit c in binding bafilomycin. PMID- 16216879 TI - A unified view of the role of electrostatic interactions in modulating the gating of Cys loop receptors. AB - In the Cys loop superfamily of ligand-gated ion channels, a global conformational change, initiated by agonist binding, results in channel opening and the passage of ions across the cell membrane. The detailed mechanism of channel gating is a subject that has lent itself to both structural and electrophysiological studies. Here we defined a gating interface that incorporates elements from the ligand binding domain and transmembrane domain previously reported as integral to proper channel gating. An overall analysis of charged residues within the gating interface across the entire superfamily showed a conserved charging pattern, although no specific interacting ion pairs were conserved. We utilized a combination of conventional mutagenesis and the high precision methodology of unnatural amino acid incorporation to study extensively the gating interface of the mouse muscle nicotinic acetylcholine receptor. We found that charge reversal, charge neutralization, and charge introduction at the gating interface are often well tolerated. Furthermore, based on our data and a reexamination of previously reported data on gamma-aminobutyric acid, type A, and glycine receptors, we concluded that the overall charging pattern of the gating interface, and not any specific pairwise electrostatic interactions, controls the gating process in the Cys loop superfamily. PMID- 16216878 TI - A novel role for glucocorticoid-induced leucine zipper protein in epithelial sodium channel-mediated sodium transport. AB - The steroid hormone aldosterone stimulates sodium (Na+) transport in tight epithelia by altering the expression of target genes that regulate the activity and trafficking of the epithelial sodium channel (ENaC). We performed microarray analysis to identify aldosterone-regulated transcripts in mammalian kidney epithelial cells (mpkC-CD(c14)). One target, glucocorticoid-induced leucine zipper protein (GILZ), was previously identified by serial analysis of gene expression (SAGE); however, its function in epithelial ion transport was unknown. Here we show that GILZ expression is rapidly stimulated by aldosterone in mpkCCD(c14) and that GILZ, in turn, strongly stimulates ENaC-mediated Na+ transport by inhibiting extracellular signal-regulated kinase (ERK) signaling. In Xenopus oocytes with activated ERK, heterologous GILZ expression consistently inhibited phospho-ERK expression and markedly stimulated ENaC-mediated Na+ current, in a manner similar to that of U0126 (a pharmacologic inhibitor of ERK signaling). In mpkCCD(c14) cells, GILZ transfection similarly consistently inhibited phospho-ERK expression and stimulated transepithelial Na+ transport. Furthermore, aldosterone treatment of mpkCCD(c14) cells suppressed phospho-ERK levels with a time course that paralleled their increase of Na+ transport. Finally, GILZ expression markedly increased cell surface ENaC expression in epidermal growth factor-treated mammalian kidney epithelial cells, HEK 293. These observations suggest a novel link between GILZ and regulation of epithelial sodium transport through modulation of ERK signaling and could represent an important pathway for mediating aldosterone actions in health and disease. PMID- 16216880 TI - Protein kinase C-alpha regulates insulin action and degradation by interacting with insulin receptor substrate-1 and 14-3-3 epsilon. AB - Protein kinase C (PKC)-alpha exerts a regulatory function on insulin action. We showed by overlay blot that PKCalpha directly binds a 180-kDa protein, corresponding to IRS-1, and a 30-kDa molecular species, identified as 14-3 3epsilon. In intact NIH-3T3 cells overexpressing insulin receptors (3T3-hIR), insulin selectively increased PKCalpha co-precipitation with IRS-1, but not with IRS-2, and with 14-3-3epsilon, but not with other 14-3-3 isoforms. Overexpression of 14-3-3epsilon in 3T3-hIR cells significantly reduced IRS-1-bound PKCalpha activity, without altering IRS-1/PKCalpha co-precipitation. 14-3-3epsilon overexpression also increased insulin-stimulated insulin receptor and IRS-1 tyrosine phosphorylation, followed by increased activation of Raf1, ERK1/2, and Akt/protein kinase B. Insulin-induced glycogen synthase activity and thymidine incorporation were also augmented. Consistently, selective depletion of 14-3 3epsilon by antisense oligonucleotides caused a 3-fold increase of IRS-1-bound PKCalpha activity and a similarly sized reduction of insulin receptor and IRS-1 tyrosine phosphorylation and signaling. In turn, selective inhibition of PKCalpha expression by antisense oligonucleotides reverted the negative effect of 14-3 3epsilon depletion on insulin signaling. Moreover, PKCalpha inhibition was accompanied by a >2-fold decrease of insulin degradation. Similar results were also obtained by overexpressing 14-3-3epsilon. Thus, in NIH-3T3 cells, insulin induces the formation of multimolecular complexes, including IRS-1, PKCalpha, and 14-3-3epsilon. The presence of 14-3-3epsilon in the complex is not necessary for IRS-1/PKCalpha interaction but modulates PKCalpha activity, thereby regulating insulin signaling and degradation. PMID- 16216881 TI - Substrate specificity and activity regulation of protein kinase MELK. AB - Maternal embryonic leucine zipper kinase (MELK) is a protein Ser/Thr kinase that has been implicated in stem cell renewal, cell cycle progression, and pre-mRNA splicing, but its substrates and regulation are not yet known. We show here that MELK has a rather broad substrate specificity and does not appear to require a specific sequence surrounding its (auto)phosphorylation sites. We have mapped no less than 16 autophosphorylation sites including serines, threonines, and a tyrosine residue and show that the phosphorylation of Thr167 and Ser171 is required for the activation of MELK. The expression of MELK activity also requires reducing agents such as dithiothreitol or reduced glutathione. Furthermore, we show that MELK is a Ca2+-binding protein and is inhibited by physiological Ca2+ concentrations. The smallest MELK fragment that was still catalytically active comprises the N-terminal catalytic domain and the flanking ubiquitin-associated domain. A C-terminal fragment of MELK functions as an autoinhibitory domain. Our data show that the activity of MELK is regulated in a complex manner and offer new perspectives for the further elucidation of its biological function. PMID- 16216882 TI - Transfer and tunneling of Ca2+ from sarcoplasmic reticulum to mitochondria in skeletal muscle. AB - The role of mitochondrial Ca2+ transport in regulating intracellular Ca2+ signaling and mitochondrial enzymes involved in energy metabolism is widely recognized in many tissues. However, the ability of skeletal muscle mitochondria to sequester Ca2+ released from the sarcoplasmic reticulum (SR) during the muscle contraction-relaxation cycle is still disputed. To assess the functional cross talk of Ca2+ between SR and mitochondria, we examined the mutual relationship connecting cytosolic and mitochondrial Ca2+ dynamics in permeabilized skeletal muscle fibers. Cytosolic and mitochondrial Ca2+ transients were recorded with digital photometry and confocal microscopy using fura-2 and mag-rhod-2, respectively. In the presence of 0.5 mM slow Ca2+ buffer (EGTA (ethylene glycolbis(2-aminoethylether)-N,N,N',N'-tetraacetic acid)), application of caffeine induced a synchronized increase in both cytosolic and mitochondrial [Ca2+]. 5 mM fast Ca2+ buffer (BAPTA (1,2-bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid)) nearly eliminated caffeine-induced increases in [Ca2+]c but only partially decreased the amplitude of mitochondrial Ca2+ transients. Confocal imaging revealed that in EGTA, almost all mitochondria picked up Ca2+ released from the SR by caffeine, whereas only about 70% of mitochondria did so in BAPTA. Taken together, these results indicated that a subpopulation of mitochondria is in close functional and presumably structural proximity to the SR, giving rise to subcellular microdomains in which Ca2+ has preferential access to the juxtaposed organelles. PMID- 16216883 TI - Diacylglycerol induces fusion of nuclear envelope membrane precursor vesicles. AB - Purified membrane vesicles isolated from sea urchin eggs form nuclear envelopes around sperm nuclei following GTP hydrolysis in the presence of cytosol. A low density subfraction of these vesicles (MV1), highly enriched in phosphatidylinositol (PtdIns), is required for nuclear envelope formation. Membrane fusion of MV1 with a second fraction that contributes most of the nuclear envelope can be initiated without GTP by an exogenous bacterial PtdIns specific phospholipase C (PI-PLC) which hydrolyzes PtdIns to form diacylglycerides and inositol 1-phosphate. This PI-PLC hydrolyzes a subset of sea urchin membrane vesicle PtdIns into diglycerides enriched in long chain, polyunsaturated species as revealed by a novel liquid chromatography-mass spectrometry analysis. Large unilammelar vesicles (LUVs) enriched in PtdIns can substitute for MV1 in PI-PLC induced nuclear envelope formation. Moreover, MV1 prehydrolyzed with PI-PLC and washed to remove inositols leads to spontaneous nuclear envelope formation with MV2 without further PI-PLC treatment. LUVs enriched in diacylglycerol mimic prehydrolyzed MV1. These results indicate that production of membrane-destabilizing diglycerides in membranes enriched in PtdIns may facilitate membrane fusion in a natural membrane system and suggest that MV1, which binds only to two places on the sperm nucleus, may initiate fusion locally. PMID- 16216884 TI - Mapping the interaction of the STT3 subunit of the oligosaccharyl transferase complex with nascent polypeptide chains. AB - Many secretory and membrane proteins are N-glycosylated by the oligosaccharyl transferase complex during their translocation across the endoplasmic reticulum membrane. Several experimental observations suggest that the highly conserved STT3 subunit contains the active site of the oligosaccharyl transferase. Here, we report a detailed study of the interaction between the active site of the STT3 protein and nascent polypeptide chains using an in vitro photocrosslinking technique. Our results show that the addition of a glycan moiety in a stretch of approximately 15 residues surrounding a QK(*)T cross-linking site impairs the interaction between the nascent chain and STT3. PMID- 16216885 TI - Determination of peptide substrate specificity for mu-calpain by a peptide library-based approach: the importance of primed side interactions. AB - Calpains are proteases that catalyze the limited cleavage of target proteins in response to Ca(2+) signaling. Because of their involvement in pathological conditions such as post-ischemic injury and Alzheimer and Parkinson disease, calpains form a class of pharmacologically significant targets for inhibition. We have determined the sequence preference for the hydrolysis of peptide substrates of the ubiquitous mu-calpain isoform by a peptide library-based approach using the proteolytic core of mu-calpain (muI-II). The approach, first described by Turk et al. (Turk, B. E., Huang, L. L., Piro, E. T., and Cantley, L. C. (2001) Nat. Biotechnol. 19, 661-667), involved the digestion of an N-terminally acetylated degenerate peptide library in conjunction with Edman sequencing to determine the specificity for residues found at primed positions. The cleavage consensus for these positions was then used to design a second, partially degenerate library, to determine specificity at unprimed positions. We have improved upon the original methodology by using a degenerate peptide dendrimer for determination of specificity at unprimed positions. By using this modified approach, the complete cleavage specificity profile for muI-II was determined for all positions flanking the cleaved peptide. A previously known preference of calpains for hydrophobic amino acids at unprimed positions was confirmed. In addition, a novel residue specificity for primed positions was revealed to highlight the importance of these sites for substrate recognition. The optimal primed site motif (MER) was shown to be capable of directing cleavage to a specific peptide bond. Accordingly, we designed a fluorescent resonance energy transfer-based substrate with optimal cleavage motifs on the primed and non primed sides (PLFAER). The mu-calpain core shows a far greater turnover rate for our substrate than for those based on the cleavage site of alpha-spectrin or the proteolytic sequence consensus compiled from substrate alignments. PMID- 16216886 TI - Essential role for CD103 in the T cell-mediated regulation of experimental colitis. AB - The integrin CD103 is highly expressed at mucosal sites, but its role in mucosal immune regulation remains poorly understood. We have analyzed the functional role of CD103 in intestinal immune regulation using the T cell transfer model of colitis. Our results show no mandatory role for CD103 expression on T cells for either the development or CD4+CD25+ regulatory T (T reg) cell-mediated control of colitis. However, wild-type CD4+CD25+ T cells were unable to prevent colitis in immune-deficient recipients lacking CD103, demonstrating a nonredundant functional role for CD103 on host cells in T reg cell-mediated intestinal immune regulation. Non-T cell expression of CD103 is restricted primarily to CD11c(high)MHC class II(high) dendritic cells (DCs). This DC population is present at a high frequency in the gut-associated lymphoid tissue and appears to mediate a distinct functional role. Thus, CD103+ DCs, but not their CD103- counterparts, promoted expression of the gut-homing receptor CCR9 on T cells. Conversely, CD103- DCs promoted the differentiation of IFN-gamma-producing T cells. Collectively, these data suggest that CD103+ and CD103- DCs represent functionally distinct subsets and that CD103 expression on DCs influences the balance between effector and regulatory T cell activity in the intestine. PMID- 16216887 TI - Nutritional stimulation of cholecystokinin receptors inhibits inflammation via the vagus nerve. AB - The immune system in vertebrates senses exogenous and endogenous danger signals by way of complex cellular and humoral processes, and responds with an inflammatory reaction to combat putative attacks. A strong protective immunity is imperative to prevent invasion of pathogens; however, equivalent responses to commensal flora and dietary components in the intestine have to be avoided. The autonomic nervous system plays an important role in sensing luminal contents in the gut by way of hard-wired connections and chemical messengers, such as cholecystokinin (CCK). Here, we report that ingestion of dietary fat stimulates CCK receptors, and leads to attenuation of the inflammatory response by way of the efferent vagus nerve and nicotinic receptors. Vagotomy and administration of antagonists for CCK and nicotinic receptors significantly blunted the inhibitory effect of high-fat enteral nutrition on hemorrhagic shock-induced tumor necrosis factor-alpha and interleukin-6 release (P < 0.05). Furthermore, the protective effect of high-fat enteral nutrition on inflammation-induced intestinal permeability was abrogated by vagotomy and administration of antagonists for CCK and nicotinic receptors. These data reveal a novel neuroimmunologic pathway, controlled by nutrition, that may help to explain the intestinal hyporesponsiveness to dietary antigens, and shed new light on the functionality of nutrition. PMID- 16216888 TI - SOCS-1 regulates IL-15-driven homeostatic proliferation of antigen-naive CD8 T cells, limiting their autoimmune potential. AB - Mice that are deficient in suppressor of cytokine signaling-1 (SOCS-1) succumb to neonatal mortality that is associated with extensive cellular infiltration of many tissues. T cells seem to be necessary for disease, which can be alleviated largely by neutralizing interferon-gamma. Examining T cell receptor (TCR) specificity shows that even monospecific T cells can mediate disease in SOCS-1 deficient mice, although disease onset is substantially faster with a polyclonal T cell repertoire. A major phenotype of SOCS-1-/- mice is the accumulation of CD44(high)CD8+ peripheral T cells. We show that SOCS-1-deficient CD8, but not CD4, T cells proliferate when transferred into normal (T cell-sufficient) mice, and that this is dependent on two signals: interleukin (IL)-15 and self-ligands that are usually only capable of stimulating homeostatic expansion in T cell deficient mice. Our findings reveal that SOCS-1 normally down-regulates the capacity of IL-15 to drive activation and proliferation of naive CD8 T cells receiving TCR survival signals from self-ligands. We show that such dysregulated proliferation impairs the deletion of a highly autoreactive subset of CD8 T cells, and increases their potential for autoimmunity. Therefore, impaired deletion of highly autoreactive CD8 T cells, together with uncontrolled activation of naive CD8 T cells by homeostatic survival ligands, may provide a basis for the T cell-mediated disease of SOCS-1-/- mice. PMID- 16216889 TI - Endogenous human cytomegalovirus gB is presented efficiently by MHC class II molecules to CD4+ CTL. AB - Human cytomegalovirus (HCMV) infects endothelial, epithelial, and glial cells in vivo. These cells can express MHC class II proteins, but are unlikely to play important roles in priming host immunity. Instead, it seems that class II presentation of endogenous HCMV antigens in these cells allows recognition of virus infection. We characterized class II presentation of HCMV glycoprotein B (gB), a membrane protein that accumulates extensively in endosomes during virus assembly. Human CD4+ T cells specific for gB were both highly abundant in blood and cytolytic in vivo. gB-specific CD4+ T cell clones recognized gB that was expressed in glial, endothelial, and epithelial cells, but not exogenous gB that was fed to these cells. Glial cells efficiently presented extremely low levels of endogenous gB--expressed by adenovirus vectors or after HCMV infection--and stimulated CD4+ T cells better than DCs that were incubated with exogenous gB. Presentation of endogenous gB required sorting of gB to endosomal compartments and processing by acidic proteases. Although presentation of cellular proteins that traffic into endosomes is well known, our observations demonstrate for the first time that a viral protein sorted to endosomes is presented exceptionally well, and can promote CD4+ T cell recognition and killing of biologically important host cells. PMID- 16216890 TI - Functional specialization of gut CD103+ dendritic cells in the regulation of tissue-selective T cell homing. AB - Gut-associated lymphoid tissue (GALT) dendritic cells (DCs) display a unique ability to generate CCR9+alpha4beta7+ gut-tropic CD8+ effector T cells. We demonstrate efficient induction of CCR9 and alpha4beta7 on CD8+ T cells in mesenteric lymph nodes (MLNs) after oral but not intraperitoneal (i.p.) antigen administration indicating differential targeting of DCs via the oral route. In vitro, lamina propria (LP)-derived DCs were more potent than MLN or Peyer's patch DCs in their ability to generate CCR9+alpha4beta7+ CD8+ T cells. The integrin alpha chain CD103 (alphaE) was expressed on almost all LP DCs, a subset of MLN DCs, but on few splenic DCs. CD103+ MLN DCs were reduced in number in CCR7-/- mice and, although CD8+ T cells proliferated in the MLNs of CCR7-/- mice after i.p. but not oral antigen administration, they failed to express CCR9 and had reduced levels of alpha4beta7. Strikingly, although CD103+ and CD103- MLN DCs were equally potent at inducing CD8+ T cell proliferation and IFN-gamma production, only CD103+ DCs were capable of generating gut-tropic CD8+ effector T cells in vitro. Collectively, these results demonstrate a unique function for LP derived CD103+ MLN DCs in the generation of gut-tropic effector T cells. PMID- 16216891 TI - Actin and agonist MHC-peptide complex-dependent T cell receptor microclusters as scaffolds for signaling. AB - T cell receptor (TCR) microclusters form within seconds of T cell contact with supported planar bilayers containing intercellular adhesion molecule-1 and agonist major histocompatibility complex (MHC)-peptide complexes, and elevation of cytoplasmic Ca2+ is observed within seconds of the first detectable microclusters. At 0-30 s after contact, TCR microclusters are colocalized with activated forms of Lck, ZAP-70, and the linker for activation of T cells. By 2 min, activated kinases are reduced in the older central microclusters, but are abundant in younger peripheral microclusters. By 5 min, TCR in the central supramolecular activation cluster have reduced activated kinases, whereas faint peripheral TCR microclusters efficiently generated activated Lck and ZAP-70. TCR microcluster formation is resistant to inhibition by Src family kinase inhibitor PP2, but is abrogated by actin polymerization inhibitor latrunculin A. We propose that Src kinase-independent formation of TCR microclusters in response to agonist MHC-peptide provides an actin-dependent scaffold for signal amplification. PMID- 16216892 TI - The shortage of drugs to treat pediatric malignancies: why does this happen? PMID- 16216893 TI - A review of qualitative research on the childhood cancer experience from the perspective of siblings: a need to give them a voice. AB - Researchers are increasingly adopting the qualitative research paradigm to study the world of siblings of children with cancer. The purpose of this review article is to identify the contributions of qualitative research in advancing the understanding of the childhood cancer experience from the perspective of siblings. Articles were selected for inclusion if (1) written in English; (2) published in a peer-reviewed journal between 1979 and present or, if a doctoral dissertation, easily accessible to the authors; (4) cited a specific qualitative research design or some type of qualitative research method of data collection and analysis; and (4) involved siblings of children with cancer as sole research participants or as research participants within the context of the family. Twenty seven qualitative research studies were reviewed. Three themes emerged from an analysis of the primary findings: (1) changing lives, (2) intense feelings, and (3) unmet needs. Limitations in the conceptualization, research design, and research methods of the qualitative studies are discussed. Suggestions that will help researchers in conducting qualitative research with siblings are also presented. PMID- 16216894 TI - Having a child diagnosed with cancer: an assessment of values from the mother's viewpoint. AB - The purpose of this qualitative study was to explore the mother's experience of having a child diagnosed with cancer. Semistructured interview questions, focused specifically on values, provided the foundation for the study. Each of the 9 participants was a mother of a child diagnosed with cancer 30 days prior to participation. Subsequent data were collected from each participant 6 months after the original interview. Using a phenomenological approach for data collection and analyses, themes were uncovered from each interview data set. The initial interviews identified 3 themes: (1) problems accessing the health care system, (2) challenges of family dynamics, and (3) support structures. Each theme persisted in the second interview, with an additional theme, (4) future plans, uncovered. Data from this study may provide guidance for health care professionals who provide care to these children and their families. The importance of maintaining communication and keeping promises cannot be overstated. PMID- 16216895 TI - Bone mineral density decrements and children diagnosed with cancer. AB - This integrative literature review is focused on (1) the prevalence of bone mineral density (BMD) decrements in children treated for acute lymphoblastic leukemia (ALL), and when these decrements are observed; (2) the risk factors associated with the development of decreased BMD and resultant complications in children treated for ALL; (3) the role, if any, that corticosteroids play in decreasing BMD in children treated for ALL; (4) interventions that can potentially manage bone loss in people treated for ALL during childhood or adolescence. The results showed that people who have been diagnosed with ALL can have decrements in BMD. The etiology of BMD decrements can be attributed to multiple factors including genetic endowment, lifestyle behaviors, the leukemia disease process, and treatment exposures especially to corticosteroids and cranial radiotherapy. Male gender, age greater than 10 years, and physical inactivity are associated with BMD decrements in ALL survivors. The role of pediatric oncology nurses in the management of bone disease in children with cancer across the illness trajectory is discussed. PMID- 16216896 TI - Treatment approaches for metastatic Ewing's sarcoma: a review of the literature. AB - The Ewing's sarcoma family of tumors (ESFT) is an aggressive group of neoplasms that represent approximately 3% of all pediatric malignancies. The overall survival rates in patients with localized disease are approaching 75%. The outcome for the 25% of patients who present with metastatic disease, however, remains poor, with long-term survival rates of less than 30%. This review will explore the natural history of ESFT including clinical presentation, molecular pathology, and high-risk features of the disease. Outcomes of metastatic treatment protocols to date will be examined as well as the rationale for current and future therapies. Nursing considerations in caring for patients with metastatic ESFT will be discussed. A case scenario will be reviewed to highlight treatment and supportive care issues in the management of the disease. Cancer therapy in general is becoming more complex; treatment approaches involve different ways of targeting tumor cells. It is crucial that nurses caring for these patients understand the rationale behind treatment strategies so that appropriate patient education and support may be given. PMID- 16216897 TI - An innovative approach to the care of patients on phase I and phase II clinical trials: the role of the experimental therapeutics nurse. AB - The tremendous strides in survival rates for childhood malignancies in large part can be attributed to the clinical trial mechanism. New and innovative therapies are being developed in the laboratory in an attempt to find a cure for those children who have relapsed or have refractory disease. Phase I and phase II clinical trials move this science from the laboratory to the patient's bedside. With increasing frequency, the oncology staff nurse may be managing the care of a patient receiving a phase I or phase II study drug. Administration of these agents goes beyond what is familiar, requires specialized knowledge, and demands a skill set beyond what is required for standard oncology care. At The Children's Hospital of Philadelphia, the role of the experimental therapeutics nurse was created in an effort to improve the process for identification, treatment, and follow-up of patients receiving these therapies. The broader role of nursing in clinical trials, the multidisciplinary challenges of experimental therapies, and the development of an innovative approach to caring for patients on phase I/II studies are discussed. PMID- 16216898 TI - Mothers' home health care work when their children have cancer. AB - The lack of research devoted to understanding the specific home health care work that mothers of children with cancer perform necessitates research in the area. This study examines the health care activities that mothers of children with cancer must assume in the home (eg, nursing care such as the administration of medicines, protecting the immune-suppressed child from concurrent illness, watching for side effects, record keeping, scheduling checkups and treatments). The implications for public policy of the extent and nature of home and lay health care in the hospital, clinic, and home are discussed. PMID- 16216900 TI - Do pacifiers reduce the risk of sudden infant death syndrome? A meta-analysis. AB - OBJECTIVE: Pacifier use has been reported to be associated with a reduced risk of sudden infant death syndrome (SIDS), but most countries around the world, including the United States, have been reluctant to recommend the use of pacifiers because of concerns about possible adverse effects. This meta-analysis was undertaken to quantify and evaluate the protective effect of pacifiers against SIDS and to make a recommendation on the use of pacifiers to prevent SIDS. METHODS: We searched the Medline database (January 1966 to May 2004) to collect data on pacifier use and its association with SIDS, morbidity, or other adverse effects. The search strategy included published articles in English with the Medical Subject Headings terms "sudden infant death syndrome" and "pacifier" and the keywords "dummy" and "soother." Combining searches resulted in 384 abstracts, which were all read and evaluated for inclusion. For the meta analysis, articles with data on the relationship between pacifier use and SIDS risk were limited to published original case-control studies, because no prospective observational reports were found; 9 articles met these criteria. Two independent reviewers evaluated each study on the basis of the 6 criteria developed by the American Academy of Pediatrics Task Force on Infant Positioning and SIDS; in cases of disagreement, a third reviewer evaluated the study, and a consensus opinion was reached. We developed a script to calculate the summary odds ratio (SOR) by using the reported ORs and respective confidence intervals (CI) to weight the ORs. We then pooled them together to compute the SOR. We performed the Breslow-Day test for homogeneity of ORs, Cochran-Mantel-Haenszel test for the null hypothesis of no effect (OR = 1), and the Mantel-Haenszel common OR estimate. The consistency of findings was evaluated and the overall potential benefits of pacifier use were weighed against the potential risks. Our recommendation is based on the taxonomy of the 5-point (A-E) scale adopted by the US Preventive Services Task Force. RESULTS: Seven studies were included in the meta-analysis. The SOR calculated for usual pacifier use (with univariate ORs) is 0.90 (95% confidence interval [CI]: 0.79-1.03) and 0.71 (95% CI: 0.59-0.85) with multivariate ORs. For pacifier use during last sleep, the SORs calculated using univariate and multivariate ORs are 0.47 (95% CI: 0.40-0.55) and 0.39 (95% CI: 0.31-0.50), respectively. CONCLUSIONS: Published case-control studies demonstrate a significant reduced risk of SIDS with pacifier use, particularly when placed for sleep. Encouraging pacifier use is likely to be beneficial on a population wide basis: 1 SIDS death could be prevented for every 2733 (95% CI: 2416-3334) infants who use a pacifier when placed for sleep (number needed to treat), based on the US SIDS rate and the last-sleep multivariate SOR resulting from this analysis. Therefore, we recommend that pacifiers be offered to infants as a potential method to reduce the risk of SIDS. The pacifier should be offered to the infant when being placed for all sleep episodes, including daytime naps and nighttime sleeps. This is a US Preventive Services Task Force level B strength of recommendation based on the consistency of findings and the likelihood that the beneficial effects will outweigh any potential negative effects. In consideration of potential adverse effects, we recommend pacifier use for infants up to 1 year of age, which includes the peak ages for SIDS risk and the period in which the infant's need for sucking is highest. For breastfed infants, pacifiers should be introduced after breastfeeding has been well established. PMID- 16216901 TI - The changing concept of sudden infant death syndrome: diagnostic coding shifts, controversies regarding the sleeping environment, and new variables to consider in reducing risk. AB - There has been a major decrease in the incidence of sudden infant death syndrome (SIDS) since the American Academy of Pediatrics (AAP) released its recommendation in 1992 that infants be placed down for sleep in a nonprone position. Although the SIDS rate continues to fall, some of the recent decrease of the last several years may be a result of coding shifts to other causes of unexpected infant deaths. Since the AAP published its last statement on SIDS in 2000, several issues have become relevant, including the significant risk of side sleeping position; the AAP no longer recognizes side sleeping as a reasonable alternative to fully supine sleeping. The AAP also stresses the need to avoid redundant soft bedding and soft objects in the infant's sleeping environment, the hazards of adults sleeping with an infant in the same bed, the SIDS risk reduction associated with having infants sleep in the same room as adults and with using pacifiers at the time of sleep, the importance of educating secondary caregivers and neonatology practitioners on the importance of "back to sleep," and strategies to reduce the incidence of positional plagiocephaly associated with supine positioning. This statement reviews the evidence associated with these and other SIDS-related issues and proposes new recommendations for further reducing SIDS risk. PMID- 16216902 TI - Differential expression of endothelin-2 along the mouse intestinal tract. AB - Endothelin (ET)-2, an ET family peptide, is highly expressed in intestine. However, the specific distribution and function of ET-2 remain unknown. We elucidated the expression profile and localization of ET-2 in mouse gastrointestinal tract. Real-time PCR analysis revealed that ET-2 gene expression in the gastrointestinal tract of healthy animals was relatively high in the colon. Immunohistochemical analysis revealed ET-2-like immunoreactivity mainly in epithelial cells of the mucosa throughout the intestinal tract of healthy animals. Intracellularly, ET-2 was concentrated close to the basement membrane of intestinal epithelial cells. A weak ET-2-like immunoreactivity was also localized to some neurofibers and the myenteric plexus of the muscle layer, coexpressing with vasoactive intestinal peptide. ET-2-like immunoreactivity was also detected at Brunner's glands of the duodenum and follicle-associated epithelium of Peyer's patch. In contrast, ET-1-like immunoreactivity was uniformly distributed in epithelial cells. In dextran sulfate sodium (DSS)-induced colitis, colonic ET-2 was upregulated during the late stage of DSS treatment. These results suggest that in intestinal epithelial cells ET-2 could be secreted into the lamina propria and the dome region in Peyer's patch, and that it might modulate immune cells in these sites for mucosal defense. PMID- 16216903 TI - Conformational dynamics of estrogen receptors alpha and beta as revealed by intrinsic tryptophan fluorescence and circular dichroism. AB - Estrogen receptors (ER alpha and ER beta) are ligand-activated nuclear receptors that mediate the action of estrogens. These receptors activate transcription by similar mechanism(s), although the overall amino acid sequence identity is only 47%. In order to compare the structural and conformational features of ER alpha and ER beta, we monitored their intrinsic tryptophan fluorescence during thermal unfolding. The 50% unfolding temperatures (T(M)) of ER alpha and ER beta were 39+/-1 and 40+/-2 degrees C, respectively. Estradiol had no significant effect on the T(M) of ER alpha or ER beta. In contrast, binding of the estrogen-response element increased the T(M) of ER alpha and ER beta by 10 degrees C. Thermal unfolding of estradiol-bound ER alpha and ligand-free ER beta showed two-step transitions, with the formation of intermediates that were stable between 36-48 and 34-42 degrees C, respectively. We confirmed the presence of intermediate states during thermal unfolding by circular dichroism spectroscopy. Atomic force microscopy showed that the ER beta intermediate consisted of discrete globular particles, whereas the ER alpha intermediate showed a speckled appearance, with sparse well-defined particles. Fluorescence-quenching studies showed the presence of two classes of tryptophan in unliganded ER alpha and ER beta. Binding of estradiol to ER beta exposed its tryptophans, whereas estradiol reduced the accessibility of the tryptophans of ER alpha. Our results illustrate the differential effects of ligands on the unfolding of ER alpha and ER beta, and identify partially unfolded intermediates. Differences in the conformational flexibility and stability of ER alpha and ER beta may represent functional differences of ligand-bound ERs in recruiting coactivator proteins and initiating transcription. PMID- 16216904 TI - ERTargetDB: an integral information resource of transcription regulation of estrogen receptor target genes. AB - The estrogen receptor (ER) plays an important role in several physiologic functions of both the reproductive and non-reproductive systems. Malignancies of the ER have been associated with the development of cancers, including those of the prostate and breast. Hence it has become of significant importance to characterize the transcriptional regulation of ER target genes. We have created ERTargetDB in order to integrate the previously published ER target gene information that is available in various publications and databases. This information resource provides researchers with an easy access to ER target genes and the regulatory mechanisms in the corresponding promoters. The current version contains 40 genes with experimentally verified estrogen response elements (EREs), 32 experimentally verified ERE tethering sites, 40 genes identified by the chromatin immunoprecipitation microarray, 381 genes from gene expression microarray and 2948 genes from computational prediction. ERTargetDB provides an integral information resource for direct target genes of ERs for the endocrinology research community. It should prove useful in the investigation of gene regulation and aid the development of computational tools for the prediction of ER target genes. PMID- 16216906 TI - Antiestrogens upregulate estrogen receptor beta expression and inhibit adrenocortical H295R cell proliferation. AB - The molecular mechanisms involved in adrenocortical tumorigenesis are still not completely understood. In this study, using the H295R cell line as a model system, we investigated the role of estrogens and estrogen receptor (ER) alpha and ER beta in the growth regulation of adrenocortical tumors. We demonstrated that H295R cells are able to convert androgens to estrogens by a constitutive expression of active cytochrome P450 aromatase protein and express ER beta to a greater extent than ER alpha. Moreover, physiological concentrations of 17beta estradiol (E2) determined an increase of thymidine incorporation, suggesting the presence of an autocrine mechanism in maintaining H295R cell proliferation. Evaluating the response to ER antagonists like 4-hydroxytamoxifen (OHT) and ICI 182 780 (ICI), we observed an up-regulation of ER beta and a dose-dependent inhibition of H295R cell proliferation. Whereas ICI determined the growth arrest of H295R cells, OHT induced morphological changes that were characteristic of apoptosis. According to the above-mentioned observations, OHT but not ICI clearly induced a marked expression of FasL and the cleavage of both caspase-8 and caspase-3. Interestingly, the apoptotic effects of OHT in H295R cells may be consequent to the enhanced levels of ER beta which stimulate the expression of FasL interacting with activating protein (AP)-1 sites located within its promoter sequence. In conclusion, we have demonstrated that H295R cells are able to transform androgens to estrogens that activate an autocrine mechanism, mediated by their own receptors, and contribute to regulate the proliferation of these cells. Moreover, this study points towards a role for ER beta as an important mediator of the repressive effects exerted by antiestrogens on H295R cells; however, further studies are needed to clarify its role in the control of adrenocortical cell proliferation and on the potential benefits of antiestrogens for treatment of adrenocortical cancer. PMID- 16216905 TI - Regulation of pancreatic beta-cell mass and proliferation by SOCS-3. AB - Growth hormone and prolactin are important growth factors for pancreatic beta cells. The effects exerted by these hormones on proliferation and on insulin synthesis and secretion in beta-cells are largely mediated through the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway. Suppressors of cytokine signaling (SOCS) proteins are specific inhibitors of the JAK/STAT pathway acting through a negative-feedback loop. To investigate in vivo effects of SOCS-3 in growth hormone (GH)/prolactin signaling in beta-cells we generated transgenic mice with beta-cell-specific overexpression of SOCS-3. The relative beta-cell proliferation and volume in the mice were measured by morphometry. Beta-cell volume of transgenic female mice was reduced by over 30% compared with beta-cell volume in wild-type female mice. Stimulation of transgenic islets in vitro with GH showed a reduced tyrosine phosphorylation of STAT-5 when compared with wild-type islets. Transduction of primary islet cultures with adenoviruses expressing various SOCS proteins followed by stimulation with GH or glucagon-like peptide-1 (GLP-1) revealed that SOCS-3 inhibited GH- but not GLP-1-mediated islet cell proliferation, indicating that the decreased beta-cell volume observed in female transgenic mice could be caused by an inhibition of GH-induced beta-cell proliferation by SOCS-3. In spite of the reduced beta-cell volume the transgenic female mice exhibited enhanced glucose tolerance compared with wild-type littermates following an oral glucose-tolerance test. Together these data suggest that SOCS-3 modulates cytokine signaling in pancreatic beta-cells and therefore potentially could be a candidate target for development of new treatment strategies for diabetes. PMID- 16216908 TI - The red wine phenolics piceatannol and myricetin act as agonists for estrogen receptor alpha in human breast cancer cells. AB - Previous epidemiological reports have suggested that red wine intake is associated with beneficial health effects due to the ability of certain phytochemical components to exert estrogen-like activity. It has been also documented that estrogens induce the proliferation of hormone-dependent breast cancer cells by binding to and transactivating estrogen receptor (ER) alpha, which in turn interacts with responsive DNA sequences located within the promoter region of target genes. In order to provide further insight into the positive association between wine consumption and the incidence of breast carcinoma in postmenopausal women, we have evaluated the estrogenic properties of two abundant wine-derived compounds, named piceatannol (PIC) and myricetin (MYR), using as model systems the hormone-sensitive MCF7 and the endocrine-independent SKBR3 breast cancer cells. On the basis of our experimental evidence PIC and MYR may contribute to the estrogenicity of red wine since: (1) they transactivate endogenous ER alpha; (2) they activate the agonist-dependent activation function (AF) 2 of ER alpha and ER beta in the context of the Gal4 chimeric proteins; (3) they rapidly induce the nuclear immunodetection of ER alpha; (4) they regulate the expression of diverse estrogen target genes; (5) they compete with 17beta estradiol for binding to ER alpha and ER beta; and--as a biological counterpart of the aforementioned abilities--(6) they exert stimulatory effects on the proliferation of MCF7 cells. Hence, the estrogenic activity of PIC and MYR might be considered at least as a potential factor in the association of red wine intake and breast tumors, particularly in postmenopausal women. PMID- 16216907 TI - Estrogen receptor alpha mediates neuronal differentiation and neuroprotection in PC12 cells: critical role of the A/B domain of the receptor. AB - Numerous studies, both in vivo and in vitro, have reported neuronal differentiating and neuroprotective actions of estrogens. Most of these estrogenic effects are mediated through specific receptors termed estrogen receptors. The aim of this study was to assess the importance of the N-terminal A/B domain of the estrogen receptor-alpha (ER alpha) in its neuronal aspects. Consequently, estrogen effects on (i) the transcriptional activity of target genes, (ii) neuronal differentiation and (iii) neuroprotection in PC12 cells transfected with either a full length form of ER alpha or an A/B domain truncated form (ER alphaCF), have been studied. We demonstrate that the maximal estrogen induced transcriptional activity of reporter genes requires a full length ER alpha, especially when cells are differentiated. Precisely, the transcriptional activity of ER alpha in differentiated cells relies, predominantly, on the activation function AF-1, located in the A/B domain. Furthermore, in PC12 cells stably expressing ER alpha, 17beta-estradiol markedly enhances the neurite outgrowth triggered by treatment with nerve growth factor and protects cells from oxidative shocks induced by depletion of glutathione. These estrogenic effects are not observed in non-transfected cells and in cells transfected with the truncated ER, devoid of the A/B domain. Altogether, these results underline the importance of the A/B domain of ER alpha in both the differentiating and the neuroprotective effects of estrogens. PMID- 16216909 TI - Structure of the glucocorticoid receptor (NR3C1) gene 5' untranslated region: identification, and tissue distribution of multiple new human exon 1. AB - The 5' untranslated region (UTR) of the glucocorticoid receptor (GR) plays a key role in determining tissue-specific expression and protein isoforms. Analysis of the 5' UTR of the human GR (hGR) has revealed 11 splice variants of the hGR exon 1, based on seven exon 1s, four of which (1-D to 1-F and 1-H) were previously unknown. All of the exon 1 variants have unique splice donor sites and share a common exon 2 splice acceptor site. Due to an upstream in-frame TGA stop codon the predicted translation from all splice variants is identical. The four new exon 1s show remarkable similarity with their rat homologues. Exon 1-D starts and finishes 17 and 36 bp upstream of the corresponding ends of the rat exon 1(4). Exon 1-E is only 6 bp longer than its homologue exon 1(5). Exon 1-F contains two short inserts of 11 and 6 bp when compared with the rat 1(7). 1-H is 18 bp longer than the corresponding rat 1(11). In addition to these new exons, we found that the human exon 1-C occurs as three distinct splice variants, covering the region homologous to the rat exons 1(9) and 1(10). All of the alternative hGR exons 1s presented here were found to be transcribed in human tissue. The human hippocampus expresses mRNA of all the exon 1 variants, while the expression of the other exon 1s seems to be tissue specific. While exon 1-D is only in the hippocampus, exons 1-E and 1-F are also detected in the immune system, and exon 1 H additionally in the liver, lung and smooth muscle. The 5' region of the hGR is more complex than previously thought, and we suggest that each of these untranslated first exons have a distinct proximal promoter region, providing additional depth to the mechanisms available for tissue-specific expression of the hGR isoforms. PMID- 16216911 TI - Androgen metabolism via 17beta-hydroxysteroid dehydrogenase type 3 in mammalian and non-mammalian vertebrates: comparison of the human and the zebrafish enzyme. AB - Formation and inactivation of testosterone is performed by various members of the 17beta-hydroxysteroid dehydrogenase (17beta-HSD) family. The main player in testosterone formation is considered to be 17beta-HSD type 3, which catalyzes the reduction of androstenedione to testosterone with high efficiency and is almost exclusively expressed in testis. So far, only the mammalian homologs have been characterized but nothing is known about the role of 17beta-HSD type 3 in other vertebrates. In this study, we describe the identification and characterization of the zebrafish homolog. We found zebrafish 17beta-HSD type 3 to be expressed in embryogenesis from sphere to 84 h post-fertilization. Expression was also detected in various tissues of both male and female adults, but displayed sexual dimorphism. Interestingly, expression was not highest in male testis but in male liver. In female adults, strongest expression was observed in ovaries. At the subcellular level, both human and zebrafish 17beta-HSD type 3 localize to the endoplasmic reticulum. The zebrafish enzyme in vitro effectively catalyzed the conversion of androstenedione to testosterone by use of NADPH as cofactor. Among further tested androgens epiandrosterone and dehydroepiandrosterone were accepted as substrates and reduced at C-17 by the human and the zebrafish enzyme. Androsterone and androstanedione though, were only substrates of human 17beta-HSD type 3, not the zebrafish enzyme. Furthermore, we found that both enzymes can reduce 11-ketoandrostenedione as well as 11beta-hydroxyandrostenedione at C-17 to the respective testosterone forms. Our results suggest that 17beta-HSD type 3 might play slightly different roles in zebrafish compared with human although testosterone itself is likely to have similar functions in both organisms. PMID- 16216910 TI - A luciferase transgenic mouse model: visualization of prostate development and its androgen responsiveness in live animals. AB - Numerous mouse models of prostate carcinogenesis have been developed, but hitherto there has been no model in which the prostate gland could be imaged in live animals. The transgenic model generated here targeted mouse prostate gland using a firefly luciferase enzyme under the control of a small but highly active and specific supra prostate-specific antigen (sPSA) promoter. We evaluated postnatal prostate development, involution and androgen-induced restoration of prostate growth in adult transgenic mice using bioluminescence imaging. Results of our study showed that: (i) the prostate gland of male offspring did not yield a significant bioluminescence signal until after sexual maturity. Luciferase was detected in the luminal epithelial cells of the ventral and dorsolateral lobes of the prostate gland and caput epididymis, with little or no activity in 18 other organs evaluated. (ii) While a constant high level of bioluminescence was detected in the mouse prostate from 5 to 35 weeks of age, a slight drop in bioluminescence was detected at 36 to 54 weeks. (iii) Upon castration, the luciferase activity signal associated with mouse prostate detected by a cooled charge-coupled device camera was dramatically reduced. This signal could be rapidly restored to pre-castration levels after androgen administration. Androgen induced luciferase activity subsided to nearly basal levels 5 days following the last injection. These data demonstrate that a bioluminescent mouse model with luciferase activity restricted to the prostate gland under the control of a (sPSA) promoter can be used on a real-time basis in live animals to investigate the development and responsiveness of the prostate gland to exogenously administered androgen. This model can be extended to detect the responsiveness of the prostate gland to therapy and used as a founder strain to visualize tumors in hosts with different genetic backgrounds. PMID- 16216912 TI - Repression of the prolactin promoter: a functional consequence of the heterodimerization between Pit-1 and Pit-1 beta. AB - The POU-homeodomain transcription factor Pit-1 is required for the differentiation of the anterior pituitary cells and the expression of their hormone products. Pit-1beta, an alternate splicing isoform, has diametrically different outcomes when it is expressed in different cell types. Pit-1beta acts as a transcriptional repressor of prolactin (PRL) and growth hormone genes in pituitary cells, and as a transcriptional activator in non-pituitary cells. In order to explore these differences, we: (1) identified the transcriptional cofactors necessary for reconstitution of repression in non-pituitary cells; (2) tested the effect of the beta-domain on heterodimerization with Pit-1 and physical interaction with the co-activator CREB binding protein (CBP); and (3) determined the beta-domain sidechain chemistry requirements for repression. Co expression of both Pit-1 isoforms reconstituted the repression of the PRL promoter in non-pituitary cells. The beta-domain allowed heterodimerization with Pit-1 but blocked physical interaction with CBP, and specific chemical properties of the beta-domain beyond hydrophobicity were dispensable. These data strongly suggest that Pit-1beta represses hormone gene expression by heterodimerizing with Pit-1 and interfering with the assembly of the Pit-1-CBP complex required for PRL promoter activity in pituitary cells. PMID- 16216913 TI - Dopamine receptor subtype 2 and somatostatin receptor subtype 5 expression influences somatostatin analogs effects on human somatotroph pituitary adenomas in vitro. AB - Dopamine (DA) and somatostatin (SRIF) receptor agonists inhibit growth hormone (GH) secretion by pituitary adenomas. We investigated DA subtype 2 receptor (DR2) and SRIF receptor (sst) subtypes 2 and 5 expression in 25 GH-secreting pituitary adenomas and tested in primary culture the effects on GH and prolactin (PRL) secretion of sst agonists selectively interacting with sst2 (BIM-23120), sst5 (BIM-23206), and sst2 and sst5 (BIM-23244). All adenomas expressed sst2; eight adenomas expressed both sst5 and DR2, eight sst5 but not DR2, and eight DR2 but not sst5. One tissue lacked expression of DR2 and sst5. GH secretion was inhibited by BIM-23120 in all samples, while it was reduced by BIM-23206 only in adenomas not expressing DR2. BIM-23120's inhibitory effects correlated with sst2 and DR2 expression, whereas DR2 expression correlated inversely with BIM-23206 inhibitory effects on GH secretion. In seven mixed GH-/PRL-secreting pituitary adenomas, PRL secretion was inhibited in sst5-expressing tumors by BIM-23206, but not by BIM-23120. BIM-23244 reduced PRL secretion only in adenomas expressing sst2, sst5 and DR2. sst5 and DR2 expression correlated directly with BIM23206 inhibitory effects on PRL secretion. Our results suggest that adenomas expressing DR2 are less likely to respond to clinically available SRIF analogs in terms of GH secretion inhibition. Therefore, drugs interacting also with DR2 might better control secretion of pituitary adenomas. PMID- 16216914 TI - ADAMTS-1 is involved in normal follicular development, ovulatory process and organization of the medullary vascular network in the ovary. AB - To clarify the role of disintegrin-like and metalloproteinase with thrombospondin type I motifs-1 (ADAMTS-1) in ovarian function, we examined abnormalities in ovulatory processes, folliculogenesis and the vascular system of ADAMTS-1 null ovaries. First, when immature female mice were treated with pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG), the number of ovulated oocytes was markedly decreased in ADAMTS-1 null mice in comparison to ADAMTS-1 (+/-) controls. The proportion of anovulated follicles to total mature follicles was significantly higher in ADAMTS-1 null females when compared with controls. The numbers of growing follicles at each stage were counted. The number of follicles at type 5b (late preantral) and later stages was markedly reduced in ADAMTS-1 null mice, irrespective of gonadotropin treatment (no gonadotropins, PMSG alone or PMSG/hCG). These data demonstrate that impairment of ovarian function to ovulate oocytes in ADAMTS-1 null mice occurs at two different levels: in the development of growing follicles and ovulatory processes. Furthermore, ADAMTS-1 null ovaries included a number of unusual atretic follicles that showed no sign of oocyte degeneration but lost the surrounding granulosa cell layers and were considered to be derived from type 4 or 5a follicles. These results suggest that ADAMTS-1 is important for follicular development beyond the type 4 and/or 5a and for maintaining normal granulosa cell layers in follicles. Finally, the number of large blood vessels in the medullar zone was significantly decreased in ADAMTS-1 null mice ovaries, suggesting that ADAMTS-1 is also involved in the organization of the medullary vascular network. PMID- 16216915 TI - Rab coupling protein (RCP): a novel target of progesterone action in primate endometrium. AB - Acquisition of functional receptivity by the endometrium is assumed to be effected by progesterone-dependent expression and repression of several genes during the implantation window in a menstrual cycle. In the present study, we employed differential display (DD) reverse transcription-polymerase chain reaction (RT-PCR) to identify progesterone-dependent gene/gene fragments that are differentially expressed during the peri-implantation phase in receptive and nonreceptive endometria, obtained from fertile and infertile bonnet monkeys respectively. Receptive endometria were obtained from regularly cycling (n=5) fertile female bonnet monkeys. Endometrial nonreceptivity was induced by treating bonnet monkeys with either 2.5 mg (n=5) or 5.0 mg (n=5) onapristone (ZK 98.299), an antiprogestin, on every third day for one cycle. Ovulation, levels of circulatory hormones (estradiol and progesterone) and menstrual cycle length did not change in treated animals; however, endometrial growth was retarded. DD2, one of the differentially expressed cDNA fragments, showed higher representation in nonreceptive endometria than in receptive endometria. The DD2 sequence was found to be homologous to the sequence of the carboxyl terminal region of Rab coupling protein (RCP), a recently discovered protein involved in intracellular vesicular trafficking. To confirm the identity of DD2 as RCP, RT-PCR studies were carried out with a forward primer deduced from the RCP sequence and a reverse primer from the DD2 sequence. The product (DDRCP) obtained, when sequenced, revealed 95% homology with the nucleotide number 1196-1757 of human RCP cDNA. Furthermore, the pattern of DDRCP expression at transcript level was found to be similar to that shown by DD2; that is, it was higher in nonreceptive endometrium. Northern analysis using labeled DD2 or DDRCP cDNA fragments identified two transcripts of 6.0 and 4.0 kb in human endometrium. In situ hybridization studies using digoxigenin-labeled DD2 revealed significantly higher (P < 0.05) localization of endometrial RCP transcripts in the proliferative phase than in the peri implantation phase in control animals. The localization was also significantly (P < 0.01) higher in peri-implantation-phase endometria from antiprogestin-treated animals than in control animals. These antiprogestin-treated animals, however, did not demonstrate any concomitant increase in the levels of immunoreactive endometrial Rab4 and Rab11 during the peri-implantation phase. A similar pattern of cycle-dependent RCP expression was observed in human endometrial biopsies. Furthermore, significantly higher (P < 0.05) levels of RCP transcripts were detected during the peri-implantation phase in women with unexplained infertility (n=3) than in fertile women (n=3). This is the first report indicating the endometrial expression of RCP and its hormonal regulation. PMID- 16216916 TI - Peroxisome proliferator-activated receptor gamma and its coactivator-1 alpha may be associated with features of the metabolic syndrome in adolescents. AB - Our objective was to search for differences in genotypes of peroxisome proliferator-activated receptor gamma (PPARgamma) (Pro12 Ala) and its coactivator PGC-1alpha (Gly482 Ser) in adolescents harboring features of metabolic syndrome. In a population-based study, we determined medical history, anthropometric variables, biochemical measurements and arterial blood pressures of 934 high school students of Caucasian origin. We selected 220 adolescents who had systolic or diastolic blood pressures more than the 80th or less than the 20th percentiles based on the previous single set of measurements. One hundred and seventy-five adolescents completed the study and underwent two additional blood pressure measurements on different days, as well as biochemical analysis and genotyping. We found no association between insulin resistance, body mass index (BMI) and leptin levels and PPARgamma and PGC-1alpha genotypes. The 12 Ala PPARgamma allele was associated with increased waist-to-hip ratio (WHR) and carriers seemed to have higher diastolic blood pressure and lower pulse pressure than non-carriers, particularly in the hypertensive and overweight group. Although Ser482 Ser PGC 1alpha homozygotes had lower WHRs than other PGC-1alpha genotypes, they were more frequent in the hypertensive group than in the normotensive (44.4 vs 24.5%, P<0.03), so the 482 Ser PGC-1 allele was in our population a risk factor for hypertension independently of WHR, homeostasis model assessment of insulin resistance, BMI and Pro12 Ala PPARgamma variant (odds ratio=4.0, 95% confidence interval 1.5-10.6, P<0.01). Multiple regression analysis showed that age- and sex adjusted systolic blood pressure correlated with the 482 Ser PGC-1 allele regardless of those covariates. In conclusion, the Gly482 Ser variant of the PGC 1alpha gene may be an independent genetic risk factor for young-onset hypertension. PMID- 16216917 TI - Hypothalamic neuropeptide expression following chronic food restriction in sedentary and wheel-running rats. AB - When rats are given access to a running-wheel in combination with food restriction, they will become hyperactive and decrease their food intake, a paradoxical phenomenon known as activity-based anorexia (ABA). Little is known about the regulation of the hypothalamic neuropeptides that are involved in the regulation of food intake and energy balance during the development of ABA. Therefore, rats were killed during the development of ABA, before they entered a state of severe starvation. Neuropeptide mRNA expression levels were analysed using quantitative real-time PCR on punches of separate hypothalamic nuclei. As is expected in a state of negative energy balance, expression levels of agouti related protein (AgRP) and neuropeptide Y (NPY) were increased 5-fold in the arcuate nucleus (ARC) of food-restricted running ABA rats vs 2-fold in sedentary food-restricted controls. The co-regulated expression of AgRP and NPY strongly correlated with relative body weight and white adipose tissue mass. Arcuate expression of pro-opiomelanocortin (POMC) and cocaine- and amphetamine-regulated transcript (CART) was reduced 2-fold in the ABA group. In second-order neurons of the lateral hypothalamic area (LHA), melanin-concentrating hormone (MCH) mRNA expression was upregulated 2-fold in food-restricted running rats, but not in food-restricted sedentary controls. Prepro-orexin, CART and corticotropin releasing hormone expression levels in the LHA and the paraventricular nucleus (PVN) were unchanged in both food-restricted groups. From this study it was concluded that during the development of ABA, neuropeptides in first-order neurons in the ARC and MCH in the LHA are regulated in an adequate response to negative energy balance, whereas expression levels of the other studied neuropeptides in secondary neurons of the LHA and PVN are unchanged and are probably regulated by factors other than energy status alone. PMID- 16216918 TI - Vitamin A supplementation induces adipose tissue loss through apoptosis in lean but not in obese rats of the WNIN/Ob strain. AB - Vitamin A is a known regulator of adipose tissue growth. In this paper, we report the possible role of dietary vitamin A supplementation in the regulation of adipose tissue mass, using a novel obese rat model of the WNIN/Ob strain developed at the National Centre for Laboratory Animal Sciences of the National Institute of Nutrition, India. Twenty-four male lean and obese rats of the WNIN/Ob strain were broadly divided into two groups at 7 months of age; each group was subdivided into two subgroups consisting of six lean and six obese rats and they were given diets containing either 2.6 mg or 129 mg vitamin A/kg diet for 2 months. Feeding a high but non-toxic dose of vitamin A (129 mg/kg diet) resulted in a significant reduction in the adiposity index and retroperitoneal white adipose tissue (RPWAT) weight in obese rats while a marginal reduction was observed in lean rats. Further, this treatment resulted in a significantly increased RPWAT apoptotic index and Bax protein expression and a decreased expression of Bcl2 in the lean rats. However, no such changes were observed in the RPWAT of the obese rats subjected to identical treatment. Thus, our data suggests that chronic dietary vitamin A supplementation at a high dose effectively regulates adipose tissue mass both in the lean and obese phenotypes of the WNIN/Ob rat strain, perhaps through different mechanisms. PMID- 16216919 TI - Independent origin of the growth hormone gene family in New World monkeys and Old World monkeys/hominoids. AB - The growth hormone (GH) gene family represents an erratic and complex evolutionary pattern, involving many evolutionary events, such as multiple gene duplications, positive selection, the birth-and-death process and gene conversions. In the present study, we cloned and sequenced GH-like genes from three species of New World monkeys (NWM). Phylogenetic analysis strongly suggest monophyly for NWM GH-like genes with respect to those of Old World monkeys (OWM) and hominoids, indicating that independent gene duplications have occurred in NWM GH-like genes. There are three main clusters of genes in putatively functional NWM GH-like genes, according to our gene tree. Comparison of the ratios of nonsynonymous and synonymous substitutions revealed that these three clusters of genes evolved under different kinds of selective pressures. Detailed analysis of the evolution of pseudogenes showed that the evolutionary pattern of this gene family in platyrrhines is in agreement with the so-called birth-and-death process. PMID- 16216920 TI - A(beta) generation in autophagic vacuoles. AB - Alzheimer's disease (AD) is the most common form of dementia among older people. It is characterized by the extracellular accumulation of beta-amyloid (Abeta) deposits called senile or neuritic plaques. Abeta is generated by the proteolytic cleavage of Abeta precursor protein (APP) by beta and gamma-secretases localized in the secretory and endocytic compartments. In this issue, Yu et al. (on p. 87) report a novel mechanism for the generation of Abeta peptides, which takes place in autophagic vacuoles (AVs) that accumulate in AD brains. PMID- 16216921 TI - Cdc25B cooperates with Cdc25A to induce mitosis but has a unique role in activating cyclin B1-Cdk1 at the centrosome. AB - Cdc25 phosphatases are essential for the activation of mitotic cyclin-Cdks, but the precise roles of the three mammalian isoforms (A, B, and C) are unclear. Using RNA interference to reduce the expression of each Cdc25 isoform in HeLa and HEK293 cells, we observed that Cdc25A and -B are both needed for mitotic entry, whereas Cdc25C alone cannot induce mitosis. We found that the G2 delay caused by small interfering RNA to Cdc25A or -B was accompanied by reduced activities of both cyclin B1-Cdk1 and cyclin A-Cdk2 complexes and a delayed accumulation of cyclin B1 protein. Further, three-dimensional time-lapse microscopy and quantification of Cdk1 phosphorylation versus cyclin B1 levels in individual cells revealed that Cdc25A and -B exert specific functions in the initiation of mitosis: Cdc25A may play a role in chromatin condensation, whereas Cdc25B specifically activates cyclin B1-Cdk1 on centrosomes. PMID- 16216922 TI - Microdomains bounded by endoplasmic reticulum segregate cell cycle calcium transients in syncytial Drosophila embryos. AB - Cell cycle calcium signals are generated by the inositol trisphosphate (InsP3) mediated release of calcium from internal stores (Ciapa, B., D. Pesando, M. Wilding, and M. Whitaker. 1994. Nature. 368:875-878; Groigno, L., and M. Whitaker. 1998. Cell. 92:193-204). The major internal calcium store is the endoplasmic reticulum (ER); thus, the spatial organization of the ER during mitosis may be important in shaping and defining calcium signals. In early Drosophila melanogaster embryos, ER surrounds the nucleus and mitotic spindle during mitosis, offering an opportunity to determine whether perinuclear localization of ER conditions calcium signaling during mitosis. We establish that the nuclear divisions in syncytial Drosophila embryos are accompanied by both cortical and nuclear localized calcium transients. Constructs that chelate InsP3 also prevent nuclear division. An analysis of nuclear calcium concentrations demonstrates that they are differentially regulated. These observations demonstrate that mitotic calcium signals in Drosophila embryos are confined to mitotic microdomains and offer an explanation for the apparent absence of detectable global calcium signals during mitosis in some cell types. PMID- 16216923 TI - Structural properties and neuronal toxicity of amyotrophic lateral sclerosis associated Cu/Zn superoxide dismutase 1 aggregates. AB - The appearance of protein aggregates is a characteristic of protein misfolding disorders including familial amyotrophic lateral sclerosis, a neurodegenerative disease caused by inherited mutations in Cu/Zn superoxide dismutase 1 (SOD1). Here, we use live cell imaging of neuronal and nonneuronal cells to show that SOD1 mutants (G85R and G93A) form an aggregate structure consisting of immobile scaffolds, through which noninteracting cellular proteins can diffuse. Hsp70 transiently interacts, in a chaperone activity-dependent manner, with these mutant SOD1 aggregate structures. In contrast, the proteasome is sequestered within the aggregate structure, an event associated with decreased degradation of a proteasomal substrate. Through the use of time-lapse microscopy of individual cells, we show that nearly all (90%) aggregate-containing cells express higher levels of mutant SOD1 and died within 48 h, whereas 70% of cells expressing a soluble mutant SOD1 survived. Our results demonstrate that SOD1 G85R and G93A mutants form a distinct class of aggregate structures in cells destined for neuronal cell death. PMID- 16216924 TI - Granuphilin molecularly docks insulin granules to the fusion machinery. AB - The Rab27a effector granuphilin is specifically localized on insulin granules and is involved in their exocytosis. Here we show that the number of insulin granules morphologically docked to the plasma membrane is markedly reduced in granuphilin deficient beta cells. Surprisingly, despite the docking defect, the exocytosis of insulin granules in response to a physiological glucose stimulus is significantly augmented, which results in increased glucose tolerance in granuphilin-null mice. The enhanced secretion in mutant beta cells is correlated with a decrease in the formation of the fusion-incompetent syntaxin-1a-Munc18-1 complex, with which granuphilin normally interacts. Furthermore, in contrast to wild-type granuphilin, its mutant that is defective in binding to syntaxin-1a fails to restore granule docking or the protein level of syntaxin-1a in granuphilin-null beta cells. Thus, granuphilin not only is essential for the docking of insulin granules but simultaneously imposes a fusion constraint on them through an interaction with the syntaxin-1a fusion machinery. These findings provide a novel paradigm for the docking machinery in regulated exocytosis. PMID- 16216925 TI - Domain III from class II fusion proteins functions as a dominant-negative inhibitor of virus membrane fusion. AB - Alphaviruses and flaviviruses infect cells through low pH-dependent membrane fusion reactions mediated by their structurally similar viral fusion proteins. During fusion, these class II viral fusion proteins trimerize and refold to form hairpin-like structures, with the domain III and stem regions folded back toward the target membrane-inserted fusion peptides. We demonstrate that exogenous domain III can function as a dominant-negative inhibitor of alphavirus and flavivirus membrane fusion and infection. Domain III binds stably to the fusion protein, thus preventing the foldback reaction and blocking the lipid mixing step of fusion. Our data reveal the existence of a relatively long-lived core trimer intermediate with which domain III interacts to initiate membrane fusion. These novel inhibitors of the class II fusion proteins show cross-inhibition within the virus genus and suggest that the domain III-core trimer interaction can serve as a new target for the development of antiviral reagents. PMID- 16216926 TI - A WASp-binding type II phosphatidylinositol 4-kinase required for actin polymerization-driven endosome motility. AB - Endosomes in yeast have been hypothesized to move through the cytoplasm by the momentum gained after actin polymerization has driven endosome abscision from the plasma membrane. Alternatively, after abscission, ongoing actin polymerization on endosomes could power transport. Here, we tested these hypotheses by showing that the Arp2/3 complex activation domain (WCA) of Las17 (Wiskott-Aldrich syndrome protein [WASp] homologue) fused to an endocytic cargo protein (Ste2) rescued endosome motility in las17DeltaWCA mutants, and that capping actin filament barbed ends inhibited endosome motility but not endocytic internalization. Motility therefore requires continual actin polymerization on endosomes. We also explored how Las17 is regulated. Endosome motility required the Las17-binding protein Lsb6, a type II phosphatidylinositol 4-kinase. Catalytically inactive Lsb6 interacted with Las17 and promoted endosome motility. Lsb6 therefore is a novel regulator of Las17 that mediates endosome motility independent of phosphatidylinositol 4-phosphate synthesis. Mammalian type II phosphatidylinositol 4-kinases may regulate WASp proteins and endosome motility. PMID- 16216927 TI - Light-regulated interaction of Dmoesin with TRP and TRPL channels is required for maintenance of photoreceptors. AB - Recent studies in Drosophila melanogaster retina indicate that absorption of light causes the translocation of signaling molecules and actin from the photoreceptor's signaling membrane to the cytosol, but the underlying mechanisms are not fully understood. As ezrin-radixin-moesin (ERM) proteins are known to regulate actin-membrane interactions in a signal-dependent manner, we analyzed the role of Dmoesin, the unique D. melanogaster ERM, in response to light. We report that the illumination of dark-raised flies triggers the dissociation of Dmoesin from the light-sensitive transient receptor potential (TRP) and TRP-like channels, followed by the migration of Dmoesin from the membrane to the cytoplasm. Furthermore, we show that light-activated migration of Dmoesin results from the dephosphorylation of a conserved threonine in Dmoesin. The expression of a Dmoesin mutant form that impairs this phosphorylation inhibits Dmoesin movement and leads to light-induced retinal degeneration. Thus, our data strongly suggest that the light- and phosphorylation-dependent dynamic association of Dmoesin to membrane channels is involved in maintenance of the photoreceptor cells. PMID- 16216928 TI - Integrin-dependent actomyosin contraction regulates epithelial cell scattering. AB - The scattering of Madin-Darby canine kidney cells in vitro mimics key aspects of epithelial-mesenchymal transitions during development, carcinoma cell invasion, and metastasis. Scattering is induced by hepatocyte growth factor (HGF) and is thought to involve disruption of cadherin-dependent cell-cell junctions. Scattering is enhanced on collagen and fibronectin, as compared with laminin1, suggesting possible cross talk between integrins and cell-cell junctions. We show that HGF does not trigger any detectable decrease in E-cadherin function, but increases integrin-mediated adhesion. Time-lapse imaging suggests that tension on cell-cell junctions may disrupt cell-cell adhesion. Varying the density and type of extracellular matrix proteins shows that scattering correlates with stronger integrin adhesion and increased phosphorylation of the myosin regulatory light chain. To directly test the role of integrin-dependent traction forces, substrate compliance was varied. Rigid substrates that produce high traction forces promoted scattering, in comparison to more compliant substrates. We conclude that integrin-dependent actomyosin traction force mediates the disruption of cell-cell adhesion during epithelial cell scattering. PMID- 16216930 TI - Fish consumption and cognitive decline with age in a large community study. AB - BACKGROUND: Dietary intake of fish and the omega-3 fatty acids have been associated with lower risk of Alzheimer disease and stroke. OBJECTIVE: To examine whether intakes of fish and the omega-3 fatty acids protect against age-related cognitive decline. DESIGN: Prospective cohort study. SETTING: Geographically defined Chicago, Ill, community. PARTICIPANTS: Residents, 65 years and older, who participated in the Chicago Health and Aging Project. MAIN OUTCOME MEASURE: Change in a global cognitive score estimated from mixed models. The global score was computed by summing scores of 4 standardized tests. In-home cognitive assessments were performed 3 times over 6 years of follow-up. RESULTS: Cognitive scores declined on average at a rate of 0.04 standardized units per year (SU/y). Fish intake was associated with a slower rate of cognitive decline in mixed models adjusted for age, sex, race, education, cognitive activity, physical activity, alcohol consumption, and total energy intake. Compared with a decline rate in score of -0.100 SU/y among persons who consumed fish less than weekly, the rate was 10% slower (-0.090 SU/y) among persons who consumed 1 fish meal per week and 13% slower (-0.088 SU/y) among persons who consumed 2 or more fish meals per week. The fish association was not accounted for by cardiovascular-related conditions or fruit and vegetable consumption but was modified after adjustment for intakes of saturated, polyunsaturated, and trans fats. There was little evidence that the omega-3 polyunsaturated fatty acids were associated with cognitive change. CONCLUSIONS: Fish consumption may be associated with slower cognitive decline with age. Further study is needed to determine whether fat composition is the relevant dietary constituent. PMID- 16216931 TI - Immunotherapy for Alzheimer disease: the promise and the problem. PMID- 16216929 TI - Inhibition of cell movement and proliferation by cell-cell contact-induced interaction of Necl-5 with nectin-3. AB - Immunoglobulin-like Necl-5/Tage4/poliovirus receptor (PVR)/CD155, originally identified as the PVR, has been shown to be up-regulated in cancer cells and to enhance growth factor-induced cell movement and proliferation. In addition, Necl 5 heterophilically trans-interacts with nectin-3, a cell-cell adhesion molecule known to form adherens junctions in cooperation with cadherin. We show here that Necl-5 was down-regulated from cell surface upon cell-cell contacts in NIH3T3 cells. This down-regulation of Necl-5 was initiated by its interaction with nectin-3 and was mainly mediated by clathrin-dependent endocytosis. Then, the down-regulation of Necl-5 induced in this way reduced movement and proliferation of NIH3T3 cells. These results indicate that the down-regulation of Necl-5 induced by its interaction with nectin-3 upon cell-cell contacts may be at least one mechanism underlying contact inhibition of cell movement and proliferation. PMID- 16216932 TI - Keeping "Trk" of paraneoplastic syndromes. PMID- 16216934 TI - Therapeutic considerations for disease progression in multiple sclerosis: evidence, experience, and future expectations. AB - In the management of patients with multiple sclerosis (MS), providers are all faced with the highly formidable challenge of ascertaining whether, and to what degree, disease-modifying therapy is effective in the individual patient. While much has been learned in randomized, controlled clinical trials, we cannot simply extrapolate the outcomes of these initiatives and apply them to the care of a single patient. In the future, the application of pharmacogenetic techniques, proteomics, and microarray analysis will yield novel profiling information on individual patients that will substantially refine the specific therapeutic questions of relevance: (1) What is the best treatment for an individual patient? (2) Which patients require intensive therapeutic combination regimens to optimize control of the disease process? (3) What are the appropriate drug dosing targets for an individual patient? and (4) Which patients will be predisposed to the development of drug-related adverse events? Such data may provide a novel variable of drug responsiveness that will mandate its inclusion into the process of covariate analyses for clinical trials. PMID- 16216935 TI - Risk factors associated with beta-amyloid(1-42) immunotherapy in preimmunization gene expression patterns of blood cells. AB - BACKGROUND: A phase 2a, double-blind, placebo-controlled, multicenter study was conducted to evaluate safety, tolerability, and pilot efficacy of immunization with beta-amyloid((1-42)) in patients with Alzheimer disease. Six immunizations were planned but were halted when meningoencephalitis was recognized as an adverse event in 6% of immunized patients. OBJECTIVE: To identify biomarkers associated with both the risk of meningoencephalitis and antibody responsiveness. PARTICIPANTS: One hundred fifty-three patients with mild to moderate Alzheimer disease.Main Outcome Measure Association between response to immunization and preimmunization expression levels of 8239 messenger RNA transcripts expressed in peripheral blood mononuclear cells that had been collected at the screening visit. RESULTS: Expression patterns of genes related to apoptosis and proinflammatory pathways (tumor necrosis factor pathway in particular) were identified as biomarkers of risk for the development of meningoencephalitis. Expression patterns of genes related to protein synthesis, protein trafficking, DNA recombination, DNA repair, and cell cycle were strongly associated with IgG response to immunization. CONCLUSIONS: Candidate biomarkers associated with risk of immunotherapy-related meningoencephalitis were detected in blood collected prior to treatment. In addition, a different set of biomarkers were identified that were associated with the desired outcome of IgG response. PMID- 16216936 TI - Hyperinsulinemia provokes synchronous increases in central inflammation and beta amyloid in normal adults. AB - BACKGROUND: Inflammation has been implicated as a pathogenetic factor in Alzheimer disease, possibly via effects on beta-amyloid (Abeta). Hyperinsulinemia induces inflammation and is a risk factor for Alzheimer disease. Thus, insulin abnormalities may contribute to Alzheimer disease pathophysiology through effects on the inflammatory network. OBJECTIVES: To determine the effects of induced hyperinsulinemia with euglycemia on Abeta, transthyretin, and inflammatory markers and modulators in plasma and cerebrospinal fluid (CSF). DESIGN: Randomized crossover trial. SETTING: Veterans Affairs hospital clinical research unit. PARTICIPANTS: Sixteen healthy adults ranging from 55 to 81 years of age (mean age, 68.2 years). INTERVENTIONS: On separate mornings, fasting participants received randomized infusions of saline or insulin (1.0 mU.kg(-1).min(-1)) with variable dextrose levels to maintain euglycemia, achieving plasma insulin levels typical of insulin resistance. Plasma and CSF were collected after an approximately 105-minute infusion. MAIN OUTCOME MEASURES: Plasma and CSF levels of interleukin 1alpha, interleukin 1beta, interleukin 6, tumor necrosis factor alpha, F2-isoprostane (CSF only), Abeta, norepinephrine, transthyretin, and apolipoprotein E. RESULTS: Insulin increased CSF levels of F2-isoprostane and cytokines (both P<.01), as well as plasma and CSF levels of Abeta42 (both P<.05). The changes in CSF levels of Abeta42 were predicted by increased F2-isoprostane and cytokine levels (both P<.01) and reduced transthyretin levels (P = .02). Increased inflammation was modulated by insulin-induced changes in CSF levels of norepinephrine and apolipoprotein E (both P<.05). CONCLUSION: Moderate hyperinsulinemia can elevate inflammatory markers and Abeta42 in the periphery and the brain, thereby potentially increasing the risk of Alzheimer disease. PMID- 16216937 TI - Central obesity and the aging brain. AB - BACKGROUND: Central adiposity as an indicator of visceral fat is linked to vascular and metabolic factors that in turn are related to cognitive decline and dementia. OBJECTIVE: To determine whether larger waist-hip ratio (WHR) is associated with structural brain changes that underlie cognitive decline and dementia. DESIGN: Cross-sectional analysis of an epidemiologic cohort study of cognitive and functional decline (Sacramento Area Latino Study on Aging). SETTING: California Central Valley. PARTICIPANTS: A total of 112 individuals selected from an ongoing cohort study of 1789 older Latino individuals. Baseline anthropomorphic measures (WHR) and measurements of fasting blood glucose, cholesterol, and insulin levels and blood pressure were obtained. MAIN OUTCOME MEASURES: Baseline magnetic resonance images were analyzed quantitatively to determine the hippocampal volumes in the right and left hemispheres and rated for the percentage of white matter hyperintensities. RESULTS: Greater WHR (P = .02) and older age (P<.001) were negatively related to hippocampal volumes. The WHR and age were positively related to white matter hyperintensities (P = .02 and P = .001, respectively). A 1-SD increase in WHR was associated with a 0.2-SD decrease in hippocampal volume and a 27% increase in white matter hyperintensities. These relationships were not affected by adjustment for body mass index, total cholesterol, fasting blood glucose, and insulin levels or systolic blood pressure in the models. CONCLUSION: A larger WHR may be related to neurodegenerative, vascular, or metabolic processes that affect brain structures underlying cognitive decline and dementia. PMID- 16216938 TI - Obesity and vascular risk factors at midlife and the risk of dementia and Alzheimer disease. AB - BACKGROUND: Vascular risk factors play a role in the development of dementia, including Alzheimer disease (AD). However, little is known about the effect of body mass index and clustering of vascular risk factors on the development of dementia. OBJECTIVE: To investigate the relation between midlife body mass index and clustering of vascular risk factors and subsequent dementia and AD. DESIGN AND SETTING: Participants of the Cardiovascular Risk Factors, Aging, and Dementia (CAIDE) study were derived from random, population-based samples previously studied in a survey carried out in 1972, 1977, 1982, or 1987. After an average follow-up of 21 years, 1449 individuals (73%) aged 65 to 79 years participated in the reexamination in 1998. MAIN OUTCOME MEASURES: Dementia and AD. RESULTS: Obesity at midlife (body mass index>30 kg/m2) was associated with the risk of dementia and AD even after adjusting for sociodemographic variables (odds ratio [OR], 2.4 [95% confidence interval (CI), 1.2-5.1]). The association was somewhat modified by further adjusting for midlife blood pressure, total cholesterol level, and smoking (OR, 2.1 [95% CI, 1.0-4.6]) and also for apolipoprotein E genotype and history of vascular disorders (OR, 1.9 [95% CI, 0.8-4.6]). Midlife obesity, high total cholesterol level, and high systolic blood pressure were all significant risk factors for dementia with ORs of around 2 for each factor, and they increased the risk additively (OR, 6.2 for the combination). CONCLUSIONS: Obesity at midlife is associated with an increased risk of dementia and AD later in life. Clustering of vascular risk factors increases the risk in an additive manner. The role of weight reduction for the prevention of dementia needs to be further investigated. PMID- 16216939 TI - Central nervous system injury-induced repulsive guidance molecule expression in the adult human brain. AB - BACKGROUND: The repulsive guidance molecule (RGM) is involved in formation of the central nervous system during development by moderating the repulsion of growing axons. However, the role of RGM in adult central nervous system lesions remains to be clarified. OBJECTIVE: To identify and determine RGM expression in adult brains with focal cerebral ischemia or traumatic brain injury and in neuropathologically unaffected control brains. Patients Twenty-one brains of patients with focal cerebral ischemia, 25 brains after traumatic brain injury, and 4 control brains. Main Outcome Measure Expression of RGM as assessed by immunohistochemical analysis. RESULTS: In normal brains, RGM expression was detected on the perikarya of some neurons, choroid plexus, smooth muscle and endothelial cells, oligodendrocytes, and myelinated white matter fibers. After focal cerebral ischemia and traumatic brain injury, RGM-immunopositive cells accumulated in lesional and perilesional areas. In hemorrhagic lesions, a massive accumulation of RGM-immunopositive cells was observed. During the first week after insult, RGM expression remained confined to neurons, smooth muscle and endothelial cells, and leukocytes infiltrating the lesion. Thereafter, with maturation of the lesion, we observed RGM expression by components of the developing scar tissue, such as fibroblastoid cells, reactive astrocytes, and a pronounced extracellular RGM deposition resembling neo-laminae. CONCLUSIONS: This is the first study of RGM in the human central nervous system. Following central nervous system injury, RGM, a novel, potent axonal growth inhibitor, is present in axonal growth impediments: the mature myelin, choroid plexus, and components of the developing scar. PMID- 16216940 TI - Skin denervation in vasculitic neuropathy. AB - BACKGROUND: Skin denervation in vasculitic neuropathy has rarely been documented despite frequent manifestations of small-fiber neuropathy including reduced sensitivity and neuropathic pain. Recently, skin biopsy has been established as a new approach to diagnose small-fiber sensory neuropathy. OBJECTIVES: To investigate the pathologic features of cutaneous nerves and to evaluate inflammatory vasculopathy in the skin of patients with vasculitis. DESIGN: Case series. SETTING: National Taiwan University Hospital, Taipei. Patients Six patients with vasculitic neuropathy. INTERVENTIONS: Patients had 3-mm punch biopsy specimens taken from the distal part of the leg (without active vasculitic lesions) and a sural nerve biopsy specimen was taken in addition to detailed neurologic examinations, laboratory investigations, and nerve conduction studies. MAIN OUTCOME MEASURES: Results of nerve conduction studies, epidermal nerve fiber density studies, and immunohistochemistry. RESULTS: All 6 patients had combined large- and small-nerve-fiber involvement on the neurologic examinations. Nerve conduction studies showed a pattern of axonal neuropathy or mononeuropathy multiplex. Epidermal nerve fiber densities were significantly reduced in the skin of all patients, consistent with concomitant small-fiber neuropathies. Perivascular infiltration by T cells and macrophages was demonstrated by immunohistochemistry. All patients experienced neurologic improvement in muscle strength and alleviation of sensory symptoms after immunotherapy with corticosteroids, plasma exchange, or cyclophosphamide. CONCLUSIONS: Small diameter sensory nerves are affected in vasculitis in addition to the well-known effect of vasculitis on large-diameter nerves. Significant inflammatory vasculopathy is present in the skin despite the absence of clinically active vasculitic lesions. PMID- 16216941 TI - Small-fiber neuropathy/neuronopathy associated with celiac disease: skin biopsy findings. AB - BACKGROUND: Celiac disease (CD) is increasingly recognized in North America and is associated with a peripheral neuropathy. OBJECTIVE: To report the clinical characteristics and skin biopsy results in patients with CD and small-fiber neuropathy symptoms. DESIGN: Case series. SETTING: Academic peripheral neuropathy clinic. PATIENTS: Eight patients with CD and neuropathy symptoms. Intervention Three-millimeter punch biopsy using the panaxonal marker protein gene product 9.5 to assess epidermal nerve fiber (ENF) density and a gluten-free diet. MAIN OUTCOME MEASURE: Clinical data and ENF density. RESULTS: All patients had asymmetric numbness and paresthesias. Three had more prominent involvement of hands than feet, and 3 had facial numbness. Celiac disease was diagnosed in 5 after their neuropathy began. The following serum antibody levels were elevated: tissue transglutaminase (n = 6), IgA gliadin (n = 4), and IgG gliadin (n = 7). Results of nerve conduction studies were normal in 7 patients. One patient had mildly reduced sural amplitudes. The ENF density was reduced in 5 patients. The ENF density was at the low limit of the normal range in 3 additional patients, 2 of whom had morphologic changes in axons. Three patients had decreased ENF density at the thigh or forearm, which was more severe than at the distal leg, compatible with a non-length-dependent process. Four reported improvement with a gluten-free diet. One had no improvement after 4 months. Symptoms developed in 2 while receiving a gluten-free diet. CONCLUSIONS: Patients with CD may have a neuropathy involving small fibers, demonstrated by results of skin biopsy. The pattern of symptoms, with frequent facial involvement and a non-length-dependent pattern on skin biopsy findings, suggests a sensory ganglionopathy or an immune mediated neuropathy. Improvement of symptoms in some patients after initiating a gluten-free diet warrants further study. PMID- 16216942 TI - LAMA2 gene analysis in congenital muscular dystrophy: new mutations, prenatal diagnosis, and founder effect. AB - OBJECTIVE: To determine if laminin-alpha2 deficiency is due to mutations in the LAMA2 gene or secondary to mutations in other congenital muscular dystrophy genes. METHODS: We performed molecular analysis of LAMA2, by single-strand conformation polymorphism and sequencing, in 15 patients with undetectable or greatly reduced laminin-alpha2 expression. We also performed 4 prenatal diagnoses and investigated a founder effect. RESULTS: We found 1 known and 9 previously undescribed LAMA2 mutations spanning all protein domains. These were nonsense or frameshifts causing laminin-alpha2 absence or, in 1 case, a homozygous missense mutation producing partial protein expression and milder phenotype. LAMA2 mutations were undetected in 5 patients, in 2 of whom FKRP mutations explained the phenotype. In 3 prenatal cases, the fetus was heterozygous for the mutation of interest and pregnancy continued; in 1 case, the fetus was affected and aborted. In 2 patients, the Cys967Stop mutation and identical haplotypes flanking the LAMA2 gene indicated a founder effect. CONCLUSIONS: The clinical phenotype was severe in most patients with LAMA2 mutations and associated with undetectable protein expression. One case with no protein and another with partial expression had milder phenotypes. Typical white matter alterations on magnetic resonance imaging were found in all patients with LAMA2 mutations, supporting the utility of magnetic resonance imaging in differential diagnosis. The founder mutation (Cys967Stop) probably originated in Albania. Genetic characterization of affected families is mainly of use for prenatal diagnosis. PMID- 16216943 TI - Autosomal dominant erythermalgia associated with a novel mutation in the voltage gated sodium channel alpha subunit Nav1.7. AB - BACKGROUND: Autosomal dominant primary erythermalgia is a rare disorder characterized by recurrent attacks of red, warm, and painful hands and/or feet. OBJECTIVE: To describe the phenotypes and molecular data of a 10-member family with 5 symptomatic living patients with erythermalgia. RESULTS: The clinical phenotype of this family was featured by episodic or continuous symmetrical red swelling, irritating warmth, and burning pain of feet and lower legs provoked or aggravated by warmth and exercise, and relief was always obtained by application of cold, such as putting feet in (ice-) cold water. The symptoms in this family were only partially controlled by analgesics and sedatives. All affected family members were heterozygous for a novel mutation (S241T) of the voltage-gated sodium channel alpha subunit Nav1.7. CONCLUSION: Primary erythermalgia may be a neuropathic disorder of the small peripheral sensory and sympathetic neurons, and may be caused by hyperexcitability of Nav1.7. PMID- 16216944 TI - Interleukin 4 and interleukin 10 levels are elevated in the cerebrospinal fluid of patients with Creutzfeldt-Jakob disease. AB - BACKGROUND: In neurodegenerative diseases, increasing attention has been focused on inflammatory mediators such as pro-inflammatory and anti-inflammatory cytokines and their potential influence in the process of neurodegeneration. In prion diseases, much data has been gained on the cell culture and animal disease models level, but only limited information is available on humans affected by Creutzfeldt-Jakob disease (CJD). OBJECTIVE: To obtain data on anti-inflammatory cytokines interleukin 4 and interleukin 10 in the cerebrospinal fluid of patients with CJD, patients with other dementia, and nondemented neurological patients and controls. DESIGN: Cerebrospinal fluid samples were collected from CJD patients and control subjects, and concentrations of the anti-inflammatory cytokines interleukin 4 and interleukin 10 were determined using an enzyme-linked immunosorbent assay. PATIENTS: Cerebrospinal fluid samples from 61 patients were analyzed. The group was composed of patients with CJD (n = 20), patients with other forms of dementia (n = 10), patients with motoneuron disease (n = 6), patients with normal pressure hydrocephalus (n = 5), and control subjects (n = 20). RESULTS: Interleukin 10 levels were significantly elevated in the cerebrospinal fluid of CJD patients (median, 9.8 pg/mL). The elevation was significant to other dementia (median, 7.9 pg/mL, P<.05), motoneuron disease (median, 7.9 pg/mL, P<.05), normal pressure hydrocephalus (median, 7.0 pg/mL, P<.05), and controls (median, 1.3 pg/mL, P<.001). Levels of interleukin 4 were significantly elevated in cerebrospinal fluid of patients with CJD (median, 26.4 pg/mL) compared with control subjects (median, 6.2 pg/mL, P<.001) and patients with a motoneuron disease (median, 10.5 pg/mL, P<.001) CONCLUSIONS: Elevated levels of the anti-inflammatory cytokines interleukin 4 and interleukin 10 in cerebrospinal fluid of patients with CJD are new findings. The data of the present study provide a clue toward the possible role of cytokines as immunological modifiers in the neurodegenerative process of CJD. PMID- 16216945 TI - Delayed neurotoxicity in primary central nervous system lymphoma. AB - BACKGROUND: Treatment for primary central nervous lymphoma (PCNSL) with chemotherapy and radiotherapy has resulted in improved survival, but some patients develop neurologic deterioration that represents a treatment-related toxic effect. This delayed neurotoxicity has been poorly defined in the literature, and the underlying mechanisms are unknown. OBJECTIVE: To describe the clinical findings, time course, and pathophysiologic mechanisms associated with neurotoxicity in an attempt to generate hypotheses for future studies that address prevention and treatment of this complication of successful PCNSL therapy. DESIGN: Retrospective review. SETTING: Department of Neurology, Memorial Sloan-Kettering Cancer Center. PATIENTS: One hundred eighty-five patients treated for PCNSL, including 43 who developed neurotoxicity. MAIN OUTCOME MEASURES: Potential risk factors, clinical course, and neuropsychological, neuroimaging, and histologic findings. RESULTS: The 5-year cumulative incidence of neurotoxicity was 24%; this incidence increases over time. Neurotoxicity presented as a rapidly progressive subcortical dementia characterized by psychomotor slowing, executive and memory dysfunction, behavioral changes, gait ataxia, and incontinence. Imaging findings revealed diffuse white matter disease and cortical-subcortical atrophy. Available autopsy data showed white matter damage with gliosis, thickening of small vessels, and demyelination. Statistical analyses were performed, accounting for death as a competing risk. Older age (P = .01), mental status changes at diagnosis (P = .04), female sex (P = .05), and radiotherapy (P<.001) predicted neurotoxicity on univariate analysis, but only radiotherapy remained significant in the multivariate setting. CONCLUSION: These findings suggest that the core pathophysiologic mechanism is the interruption of frontal-subcortical circuits mediated by radiation damage, possibly caused by progressive microvascular alterations, loss of oligodendrocyte progenitors, or oxidative stress. PMID- 16216947 TI - Impairment of Trk-neurotrophin receptor by the serum of a patient with subacute sensory neuropathy. AB - BACKGROUND: Paraneoplastic peripheral neuropathy is sometimes associated with unidentified neuronal autoantibodies. OBJECTIVE: To examine the effects of serum from a patient with subacute sensory axonopathy on the function of the Trk high affinity nerve growth factor receptor. PATIENT: An 86-year-old man with sensory neuropathy exhibiting an autoantibody to Trk. METHODS: Immunoblot analyses of the brain homogenates and immunoprecipitation were performed with human sera. We further examined the effect of sera on nerve growth factor-induced neurite outgrowth and Trk autophosphorylation. RESULTS: The patient showed sensory nerve axonopathy without well-known paraneoplastic autoantibodies. His serum inhibited nerve growth factor-induced neurite outgrowth and Trk autophosphorylation in PCtrk cells. Moreover, the patient's serum, but not control serum, immunoprecipitated Trk and recognized Trk in brain homogenates as well as in Trk immunoprecipitates. CONCLUSION: These data strongly suggest that an anti-Trk autoantibody might cause subacute sensory neuropathy. PMID- 16216948 TI - Clinical stabilization and effective B-lymphocyte depletion in the cerebrospinal fluid and peripheral blood of a patient with fulminant relapsing-remitting multiple sclerosis. AB - BACKGROUND: The anti-CD20 monoclonal antibody rituximab effectively depletes B lymphocytes and has been successfully used in the therapy of immune-mediated disorders of the peripheral nervous system. A limited effect of rituximab on B lymphocytes in the cerebrospinal fluid compartment of patients with primary progressive multiple sclerosis (MS) was recently reported. OBJECTIVE: To determine the effect of rituximab on clinical, magnetic resonance imaging, and immunological variables in a patient with relapsing-remitting MS. DESIGN: A patient with relapsing-remitting MS was treated with rituximab. The patient was repeatedly examined clinically and by magnetic resonance imaging. The frequency of peripheral blood and cerebrospinal fluid B lymphocytes was assessed by flow cytometry before, during, and after rituximab therapy. RESULTS: Rituximab monotherapy resulted in significant clinical improvement. Inflammatory surrogate markers on magnetic resonance imaging were also reduced. B lymphocytes were depleted in the cerebrospinal fluid and peripheral blood. CONCLUSIONS: Our data demonstrate beneficial clinical effects of rituximab in relapsing-remitting MS, mediated through modulation of humoral systemic and central nervous system intrinsic immune responses. Clinical trials should determine optimal therapeutic strategies for patients with relapsing-remitting MS. PMID- 16216946 TI - Delusions and hallucinations are associated with worse outcome in Alzheimer disease. AB - BACKGROUND: Delusions and hallucinations are common in Alzheimer disease (AD) and there are conflicting reports regarding their ability to predict cognitive decline, functional decline, and institutionalization. According to all previous literature, they are not associated with mortality. OBJECTIVE: To examine whether the presence of delusions or hallucinations has predictive value for important outcomes in AD. DESIGN, SETTING, AND PARTICIPANTS: A total of 456 patients with AD at early stages (mean Folstein Mini-Mental State Examination [MMSE] score of 21 of 30 at entry) were recruited and followed up semiannually for up to 14 years (mean, 4.5 years) in 5 university-based AD centers in the United States and Europe. Using the Columbia University Scale for Psychopathology in AD (administered every 6 months, for a total of 3266 visit-assessments, average of 7.2 per patient), the presence of delusions and hallucinations was extracted and examined as time-dependent predictors in Cox models. The models controlled for cohort effect, recruitment center, informant status, sex, age, education, a comorbidity index, baseline cognitive and baseline functional performance, behavioral symptoms, and use of neuroleptics and cholinesterase inhibitors. MAIN OUTCOME MEASURES: Cognitive (Columbia MMSE score of < or =20/57 [approximate Folstein MMSE score of < or =10/30]), functional (Blessed Dementia Rating Scale [parts I and II] score of > or =10), institutionalization equivalent index, and death. RESULTS: During the full course of follow-up, 38% of patients reached the cognitive, 41% the functional, 54% the institutionalization, and 49% the mortality end point. Delusions were noted for 34% of patients at baseline and 70% at any evaluation. Their presence was associated with increased risk for cognitive (risk ratio [RR], 1.50; 95% confidence interval [CI], 1.07-2.08) and functional decline (RR, 1.41; 95% CI, 1.02-1.94). Hallucinations were present in 7% of patients at initial visit and in 33% at any visit. Their presence was associated with increased risk for cognitive decline (RR, 1.62; 95% CI, 1.06 2.47), functional decline (RR, 2.25; 95% CI, 1.54-2.27), institutionalization (RR, 1.60; 95% CI, 1.13-2.28), and death (RR, 1.49; 95% CI, 1.03-2.14). CONCLUSIONS: Delusions and hallucinations are very common in AD and predict cognitive and functional decline. Presence of hallucinations is also associated with institutionalization and mortality. PMID- 16216949 TI - Presenilin 1 Glu318Gly polymorphism: interpret with caution. AB - BACKGROUND: The significance of the presenilin 1 (PSEN1) Glu318Gly polymorphism has been described as either a causal mutation with reduced penetrance or a benign polymorphism. When this polymorphism is found in a symptomatic person with a family history of dementia, counseling on recurrence risk becomes very problematic. OBJECTIVE: To demonstrate that the PSEN1 Glu318Gly polymorphism should be interpreted cautiously. DESIGN: Case histories of 2 patients with presenile dementia and family histories of dementia are described. The PSEN1 gene was sequenced in the patients and in 11 family members of patient 1. RESULTS: Two patients with presenile dementia and personality change were found to carry the PSEN1 Glu318Gly polymorphism. The presence of the polymorphism was confirmed in several family members of patient 1 but was absent in 1 symptomatic relative. CONCLUSIONS: The Glu318Gly polymorphism may be associated with risk for neurodegenerative disease; however, in the cases described here, it did not appear to be a risk factor. Until there is consensus on whether it is associated with disease, families should be informed that the clinical significance of the polymorphism is uncertain. PMID- 16216950 TI - Wilson disease with an initial manifestation of polyneuropathy. AB - BACKGROUND: Recognition of Wilson disease (WD) is sometimes difficult because of its diverse manifestations. Peripheral neuropathy is rarely reported in the context of WD. OBJECTIVE: To report an unusual patient with WD whose initial manifestation was peripheral neuropathy. DESIGN: Case report. SETTING: Neurology department in a tertiary referral center. METHOD: Personal observation. RESULT: A 17-year-old man, who was eventually diagnosed with WD, was initially seen with polyneuropathy at least 6 months prior to developing more typical symptoms of WD. Electrophysiological and pathological studies suggested a neuropathy of mixed type. Treatment for WD resulted in clinical and electrophysiological improvement. CONCLUSION: Wilson disease may initially appear as a treatable polyneuropathy. PMID- 16216951 TI - Central neurogenic hyperventilation: a case report and discussion of pathophysiology. AB - BACKGROUND: Central neurogenic hyperventilation is a rare condition with poorly understood pathophysiology. OBJECTIVE: To describe a patient with central neurogenic hyperventilation caused by an infiltrative brainstem lymphoma. CONCLUSION: Based on analysis of this patient and other case reports, we propose that central neurogenic hyperventilation is uniquely the result of infiltrative tumors that stimulate pontine respiratory centers and central chemoreceptors. PMID- 16216952 TI - Intraoperative monitoring. PMID- 16216953 TI - Resolution of bilateral thalamic lesions due to deep cerebral venous thrombosis. PMID- 16216954 TI - Effective suppression of cerebrospinal fluid B cells by rituximab and cyclophosphamide in progressive multiple sclerosis. PMID- 16216955 TI - Significance of recurrent mutations in the myofibrillogenesis regulator 1 gene. PMID- 16216956 TI - Pallidal vs subthalamic deep brain stimulation for Parkinson disease: winner and loser or a sharing of honors? PMID- 16216957 TI - Multisite randomized trial of deep brain stimulation. PMID- 16216958 TI - Corin gene minor allele defined by 2 missense mutations is common in blacks and associated with high blood pressure and hypertension. AB - BACKGROUND: The natriuretic peptide system contributes to blood pressure regulation. Atrial and brain natriuretic peptides are cleaved into smaller biologically active molecules by corin, a transmembrane serine protease expressed in cardiomyocytes. METHOD AND RESULTS: This genotype-phenotype genetic association study included replication samples and genomic control to correct for population stratification. Sequencing of the human corin gene identified 2 nonsynonymous, nonconservative single nucleotide polymorphisms (Q568P and T555I) in near-complete linkage disequilibrium, thus describing a single minor I555 (P568) corin gene allele. This allele was present in the heterozygote state in &12% of blacks but was extremely rare in whites (<0.5% were homozygous for the minor allele). In our primary population sample, the Dallas Heart Study, after adjustment for potential confounders, including population stratification, the corin I555 (P568) allele remained independently associated with increased risk for prevalent hypertension (odds ratio, 1.63; 95% CI, 1.11 to 2.38; P=0.013). The corin I555 (P568) allele also was associated with higher systolic blood pressure in subjects not using antihypertensive medication in unadjusted (133.7+/-20.7 versus 129.4+/-17.4 mm Hg; P=0.029) and adjusted (132.5+/-1.6 versus 128.9+/-0.6 mm Hg; P=0.029) analyses. The independent association of the minor corin allele with increased risk for prevalent hypertension was confirmed in the Multi-Ethnic Study of Atherosclerosis (odds ratio, 1.50; 95% CI, 1.09 to 2.06; P=0.014). In addition, the association of the minor corin I555 (P568) allele with higher systolic blood pressure was confirmed in adjusted analysis in the Chicago Genetics of Hypertension Study (125.8+/-1.9 versus 121.4+/-0.7 mm Hg; P=0.03). CONCLUSIONS: The corin I555 (P568) allele is common in blacks and is associated with higher blood pressure and an increased risk for prevalent hypertension. PMID- 16216959 TI - Role of nitric oxide in mediating in vivo vascular responses to calcitonin gene related peptide in essential and peripheral circulations in the fetus. AB - BACKGROUND: The role of calcitonin gene-related peptide (CGRP) in cardiovascular regulation is gaining clinical and scientific interest. In the adult, in vivo studies have shown that CGRP-stimulated vasodilation in several vascular beds depends, at least in part, on nitric oxide (NO). However, whether CGRP acts as a vasodilator in the fetus in vivo and whether this effect is mediated via NO have been addressed only minimally. This study tested the hypothesis that CGRP has potent NO-dependent vasodilator actions in essential and peripheral vascular beds in the fetus in late gestation. METHODS AND RESULTS: Under anesthesia, 5 fetal sheep at 0.8 gestation were instrumented with vascular catheters and Transonic flow probes around an umbilical artery and a femoral artery. Five days later, fetuses received 2- and 5-microg doses of exogenous CGRP intra-arterially in randomized order. Doses were repeated during NO blockade with the NO clamp. This technique permits blockade of de novo synthesis of NO while compensating for tonic production of the gas, thereby maintaining basal cardiovascular function. CGRP resulted in potent and long-lasting NO-dependent dilation in the umbilical and femoral circulations, hypotension, and a positive cardiac chronotropic effect. During NO blockade, the femoral vasodilator response to CGRP was diminished. In contrast, in the umbilical vascular bed, the dilator response was not only prevented but reversed to vasoconstriction. CONCLUSIONS: CGRP has potent NO-dependent vasodilator actions in fetal essential and peripheral vascular beds. CGRP-induced NO-dependent effects in the umbilical vascular bed may provide an important mechanism in the control and maintenance of umbilical blood flow during pregnancy. PMID- 16216960 TI - Clinical features, management, and outcome of children with fetal and postnatal diagnoses of isomerism syndromes. AB - BACKGROUND: Isomerism is associated with a complex spectrum of anomalies. There is paucity of data on prenatally detected cases. METHODS AND RESULTS: Between January 1990 and February 2004, 83 of 166 cases (50%) had a prenatal diagnosis of left isomerism (LAI; 52 of 97) or right isomerism (RAI; 31 of 69) at our institution. The spectrum of anomalies, management, and outcomes was compared for fetal and postnatal diagnoses of LAI and RAI. RAI more often than LAI was associated with AV septal defect (90% versus 56%; P<0.0001), pulmonary outflow obstruction (91% versus 37%; P<0.0001), total anomalous pulmonary venous drainage (73% versus 13%; P<0.0001), and abnormal VA connections (68% versus 33%; P<0.0001), whereas inferior vena cava interruption (3% versus 93%; P<0.0001), complete AV block (0% versus 13%; P=0.004), aortic obstruction (6% versus 33%; P<0.0001), and extracardiac defects (5% versus 25%; P=0.006) were less common. The spectrum of lesions was comparable for fetal and postnatal cases, except for AV block (fetal, 25%; postnatal, 0%; P=0.0002) and AV septal defect (fetal, 67%; postnatal, 42%; P=0.023) in LAI. Fetal demise was due mainly to pregnancy termination (LAI, 42%; RAI, 45%). Survival of actively managed children with LAI was significantly better than for those with RAI (P<0.0001) but did not differ with regard to fetal versus postnatal diagnosis. Most LAI cases required no intervention or underwent successful biventricular cardiac surgery (65%), unlike RAI cases (13%; P<0.0001). CONCLUSIONS: Prenatal diagnosis did not affect overall survival despite facilitated care. The prognosis of RAI was worse compared with LAI because of more complex associated cardiac defects and the inability to perform successful surgical procedures. PMID- 16216961 TI - Angiotensin receptor blockade and exercise capacity in adults with systemic right ventricles: a multicenter, randomized, placebo-controlled clinical trial. AB - BACKGROUND: Pharmacological blockade of the renin-angiotensin system improves exercise tolerance in patients with left ventricular dysfunction, yet its impact on patients with systemic right ventricles (RVs) remains unknown. METHODS AND RESULTS: A multicenter, randomized, double-blind, placebo-controlled, crossover clinical trial was performed to assess the effects of losartan on exercise capacity and neurohormonal levels in patients with systemic RVs. Of 29 patients studied (age, 30.3+/-10.9 years), 21 had transposition of the great arteries with a Mustard baffle, and 8 had congenitally corrected transposition of the great arteries. Baseline values were as follows: VO2max, 29.8+/-5.6 mL.kg(-1).min(-1) (73.5+/-12.9% predicted value); RV ejection fraction, 41.6+/-9.3%; N-terminal pro brain natriuretic peptide (NT-proBNP), 257.7+/-243.4 pg/mL (normal <125 pg/mL); and angiotensin II, 5.7+/-4.9 pg/mL (normal <5.0 pg/mL). Comparing losartan to placebo showed no differences in VO2max (29.9+/-5.4 versus 29.4+/-6.2 mL.kg( 1).min(-1); P=0.43), exercise duration (632.3+/-123.0 versus 629.9+/-140.7 seconds; P=0.76), and NT-proBNP levels (201.2+/-267.8 versus 229.7+/-291.5 pg/mL; P=0.10), despite a trend toward increased angiotensin II levels (15.2+/-13.8 versus 8.8+/-12.5 pg/mL; P=0.08). CONCLUSIONS: In adults with systemic RVs, losartan did not improve exercise capacity or reduce NT-proBNP levels. Minimal baseline activation of the renin-angiotensin system may explain this lack of benefit and imply an alternative pathophysiological mechanism for the progressive ventricular dysfunction and impaired exercise capacity observed in such patients. PMID- 16216962 TI - Endogenous vascular hydrogen peroxide regulates arteriolar tension in vivo. AB - BACKGROUND: Although many studies suggested direct vasomotor effects of hydrogen peroxide (H2O2) in vitro, little is known about the vasomotor effects of H2O2 in vivo. METHODS AND RESULTS: We have generated mice overexpressing human catalase driven by the Tie-2 promoter to specifically target this transgene to the vascular tissue. Vessels of these mice (cat++) expressed significantly higher levels of catalase mRNA, protein, and activity. The overexpression was selective for vascular tissue, as evidenced by immunohistochemistry in specimens of aorta, heart, lung, and kidney. Quantification of reactive oxygen species by fluorescence signals in cat++ versus catalase-negative (catn) mice showed a strong decrease in aortic endothelium and left ventricular myocardium but not in leukocytes. Awake male cat++ at 3 to 4 months of age had a significantly lower systolic blood pressure (sBP, 102.7+/-2.2 mm Hg, n=10) compared with their transgene-negative littermates (catn, 115.6+/-2.5 mm Hg, P=0.0211) and C57BL/6 mice (118.4+/-3.06 mm Hg, n=6). Treatment with the catalase inhibitor aminotriazole increased sBP of cat++ to 117.3+/-4.3 mm Hg (P=0.0345), while having no effect in catn (118.4+/-2.4 mm Hg, n=4, P>0.05). In contrast, treatment with the NO-synthase inhibitor nitro-L-arginine methyl ester (100 mg.kg BW(-1).d( 1)) increased sBP in cat++ and C57Bl/6 to a similar extent. Likewise, phosphorylation of vasodilator-stimulated phosphoprotein in skeletal muscle, left ventricular myocardium, and lung was identical in cat++ and catn. Endothelium- and NO-dependent aortic vasodilations were unchanged in cat++. Aortic KCl contractions were significantly lower in cat++ and exogenous H2O2 (10 micromol/L) induced vasoconstriction. CONCLUSIONS: These data suggest that endogenous H2O2 may act as a vasoconstrictor in resistance vessels and contribute to the regulation of blood pressure. PMID- 16216963 TI - Angiotensin-converting enzyme inhibitors: a new mechanism of action. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors are valuable agents for the treatment of hypertension, heart failure, and other cardiovascular and renal diseases. The cardioprotective effects of ACE inhibitors are mediated by blockade of both conversion of angiotensin (Ang) I to Ang II and kinin hydrolysis. Here, we report a novel mechanism that may explain the cardiac antifibrotic effect of ACE inhibition, involving blockade of the hydrolysis of N acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP). METHODS AND RESULTS: To study the role of Ac-SDKP in the therapeutic effects of the ACE inhibitor captopril, we used a model of Ang II-induced hypertension in rats treated with the ACE inhibitor either alone or combined with a blocking monoclonal antibody (mAb) to Ac-SDKP. These hypertensive rats had left ventricular hypertrophy (LVH) as well as increases in cardiac fibrosis, cell proliferation, transforming growth factor beta (TGF-beta) expression, and phosphorylation of Smad2 (P-Smad2), a signaling mediator of the effects of TGF-beta. The ACE inhibitor did not decrease either blood pressure or LVH; however, it significantly decreased LV collagen from 13.3+/-0.9 to 9.6+/-0.6 microg/mg dry wt (P<0.006), and this effect was blocked by the mAb (12.1+/-0.6; P<0.034, ACE inhibitor versus ACE inhibitor+mAb). In addition, analysis of interstitial collagen volume fraction and perivascular collagen (picrosirius red staining) showed a very similar tendency. Likewise, the ACE inhibitor significantly decreased LV monocyte/macrophage infiltration, cell proliferation, and TGF-beta expression, and these effects were blocked by the mAb. Ang II increased Smad2 phosphorylation 3.2+/-0.9-fold; the ACE inhibitor lowered this to 0.6+/-0.1-fold (P<0.001), and the mAb blocked this decrease to 2.1+/-0.3 (P<0.001, ACE inhibitor versus ACE inhibitor+mAb). Similar findings were seen when the ACE inhibitor was replaced by Ac-SDKP. CONCLUSIONS: We concluded that in Ang II-induced hypertension, the cardiac antifibrotic effect of ACE inhibitors is a result of the inhibition of Ac-SDKP hydrolysis, resulting in a decrease in cardiac cell proliferation (probably fibroblasts), inflammatory cell infiltration, TGF-beta expression, Smad2 activation, and collagen deposition. PMID- 16216964 TI - Occult cardiac contractile dysfunction in dystrophin-deficient children revealed by cardiac magnetic resonance strain imaging. AB - BACKGROUND: Duchenne muscular dystrophy (DMD) is an inherited disease characterized by early onset of skeletal muscle degeneration and progressive weakness. Although dilated cardiomyopathy may occur during adolescence, it is often undetected early in its course because of physical inactivity and generalized debilitation. The purpose of this study was to apply the technique of cardiac magnetic resonance (CMR) tagging to detect occult cardiac dysfunction in young subjects with DMD by measuring myocardial strain and torsion. METHODS AND RESULTS: Thirteen DMD pediatric subjects without clinically apparent heart disease and 9 age-matched healthy males were recruited. Each was scanned on a 1.5 T clinical scanner to acquire contiguous short-axis planes from the apex to the mitral valve plane and then 3 tagged images at base, midventricle, and apex. Global and segmental myocardial net twist and circumferential strain were computed with the use of 2D homogeneous strain analysis. Ventricular torsion was computed by normalizing net twist by the distance from apex to mitral valve plane. DMD patients exhibited normal left ventricular volumes and ejection fractions but manifested reduced midventricular and basal cross-sectional global circumferential strain compared with the reference group (P<0.005). These alterations also appeared in segmental analyses in the septal, anterior, lateral, and inferior walls (P<0.05). CONCLUSIONS: In patients predisposed to cardiomyopathies because of dystrophinopathy, occult regional cardiac dysfunction can be diagnosed with CMR tagging. This method of strain imaging analysis may offer a sensitive approach for delineating the presence and progression of cardiovascular disease and for assessing therapies designed to modulate the onset and course of heart failure. PMID- 16216965 TI - Therapy of stable angina pectoris: the uncomplicated patient. PMID- 16216966 TI - Images in cardiovascular medicine. Traumatic pseudoaneurysm of the descending thoracic aorta. PMID- 16216967 TI - Images in cardiovascular medicine. Heart on the heart. PMID- 16216968 TI - Images in cardiovascular medicine. Giant pulmonary aneurysm associated with pulmonary valve agenesia. PMID- 16216969 TI - Letter regarding article by Kuhl et al, "High prevalence of viral genomes and multiple viral infections in the myocardium of adults with 'idiopathic' left ventricular dysfunction". PMID- 16216970 TI - Letter regarding article by Becker et al, "Hyperhomocysteinemia, a cardiac metabolic disease: role of nitric oxide and the p22phox subunit of NADPH oxidase". PMID- 16216971 TI - Sudden cardiac arrest from primary electrical diseases: provoking concealed arrhythmogenic syndromes. PMID- 16216972 TI - Anatomy of an emerging diagnostic test: computed tomographic coronary angiography. PMID- 16216973 TI - Inflammatory biomarkers for prediction of outcomes after unprotected left main coronary intervention. PMID- 16216974 TI - Myeloid differentiation factor-88 plays a crucial role in the pathogenesis of Coxsackievirus B3-induced myocarditis and influences type I interferon production. AB - BACKGROUND: Myeloid differentiation factor (MyD)-88 is a key adaptor protein that plays a major role in the innate immune pathway. How MyD88 may regulate host response in inflammatory heart disease is unknown. METHODS AND RESULTS: We found that the cardiac protein level of MyD88 was significantly increased in the hearts of wild-type mice after exposure to Coxsackievirus B3 (CVB3). MyD88(-/-) mice showed a dramatic higher survival rate (86%) in contrast to the low survival (35%) in the MyD88(+/+) mice after CVB3 infection (P<0.0001). Pathological examination showed a significant decrease of cardiac and pancreatic inflammation in the MyD88(-/-) mice. Viral concentrations in the hearts were significantly decreased in the MyD88(-/-) mice. Cardiac mRNA levels for interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and IL-18 were significantly decreased in the MyD88(-/-) mice. Similarly, serum levels of T helper 1 cytokines were significantly decreased in the MyD88(-/-) mice. In contrast, cardiac protein levels of the activated interferon regulatory factor (IRF)-3 and IFN-beta were significantly increased in the MyD88(-/-) mice but not other usual upstream signals to IRF-3. The cardiac expression of coxsackie adenoviral receptor and p56(lck) were also significantly decreased. CONCLUSIONS: MyD88 appears to be a key contributor to cardiac inflammation, mediating cytokine production and T-helper-1/2 cytokine balance, increasing coxsackie-adenoviral receptor and p56(lck) expression and viral titers after CVB3 exposure. Absence of MyD88 confers host protection possibly through novel direct activation of IRF-3 and IFN-beta. PMID- 16216975 TI - Effects of heat stress on thermoregulatory responses in congestive heart failure patients. AB - BACKGROUND: Clinical observations suggest that tolerance to heat stress may be impaired in patients with cardiovascular diseases, particularly those associated with impaired ventricular function and congestive heart failure (CHF). However, thermoregulatory function during a controlled heat stress challenge in patients with CHF has not been studied. METHODS AND RESULTS: To test the hypothesis that thermoregulatory responses are attenuated in such patients, we assessed cutaneous vasodilation and sweat rate in patients with stable class II-III CHF and in matched healthy subjects during passive whole-body heating. Whole-body heating induced a similar increase in internal temperature (approximately 0.85 degrees C) in both groups. The sweating responses in patients with CHF were not significantly different from that in control subjects. In contrast, the elevation in forearm cutaneous vascular conductance in patients with CHF was reduced by nearly 50% relative to the control subjects (3.8+/-0.8 versus 6.9+/-1.0 mL/100 mL tissue per minute per 100 mm Hg, P=0.04). Moreover, maximal cutaneous vasodilator capacity to direct local heating in patients with CHF was also significantly lower than in control subjects, suggesting that vascular remodeling may be limiting cutaneous vasodilation during hyperthermia. CONCLUSIONS: These observations suggest that patients with CHF exhibit attenuated cutaneous vasodilator responses to both whole-body and local heating, whereas sweating responses are preserved. Attenuated cutaneous vasodilation may be a potential mechanism for heat intolerance in patients with CHF. PMID- 16216976 TI - Mechanoreflex mediates the exaggerated exercise pressor reflex in heart failure. AB - BACKGROUND: In heart failure, exercise elicits excessive increases in mean arterial pressure (MAP) and heart rate (HR). Using a novel rat model, we previously demonstrated that this exaggerated cardiovascular responsiveness is mediated by an overactive exercise pressor reflex (EPR). Although we previously determined that abnormalities in the group IV afferent neuron population (associated with the metabolic component of the reflex) initiate the development of the exaggerated EPR in heart failure, these fibers do not mediate the enhanced circulatory responses to exercise. Therefore, we hypothesized that the augmentation in EPR activity is primarily mediated by the mechanically sensitive component of the reflex (mediated predominately by activation of group III afferent fibers). METHODS AND RESULTS: Male Sprague-Dawley rats were divided into 3 groups: sham (control), dilated cardiomyopathic (DCM), and neonatal capsaicin treated animals (NNCAP, group IV afferent fibers ablated). Activation of the EPR by electrically induced static muscle contraction of the hindlimb resulted in larger increases in MAP and HR in DCM and NNCAP compared with sham animals. In all groups, administration of gadolinium (a selective blocker of mechanically sensitive receptors) within the hindlimb attenuated the MAP and HR responses to contraction. However, the magnitude of this reduction was greater in DCM and NNCAP compared with sham animals. CONCLUSIONS: From these data, we conclude that the muscle mechanoreflex mediates the exaggerated EPR that develops in heart failure. Moreover, these findings suggest that mechanoreflex overactivity in heart failure may be a compensatory response to functional alterations in group IV fibers. Given these findings, the muscle mechanoreflex may serve as a novel target in the treatment of the abnormal circulatory responses to exercise in heart failure. PMID- 16216977 TI - Headaches and the treatment of blood pressure: results from a meta-analysis of 94 randomized placebo-controlled trials with 24,000 participants. AB - BACKGROUND: Uncertainty exists over whether blood pressure-lowering drugs prevent headache. METHODS AND RESULTS: A meta-analysis was carried out of the 94 randomized placebo-controlled trials of 4 different classes of blood pressure lowering drugs (thiazides, beta-blockers, ACE inhibitors, and angiotensin II receptor antagonists) in fixed doses in which data on headache were reported. There were 17,641 participants who were allocated blood pressure-lowering drugs and 6603 who were allocated placebo. Treatment lowered systolic and diastolic blood pressures by 9.4 and 5.5 mm Hg, respectively, on average. One third fewer people on average reported headache in the treated groups (8.0%) than the placebo groups (12.4%) (odds ratio, 0.67; 95% CI, 0.61 to 0.74; P<0.001). About 1 in 30 treated persons benefited by having headache prevented. The prevalence of headache was reduced (P<0.001) in trials of each of the 4 classes of drugs. CONCLUSIONS: Our results show that blood pressure-lowering drugs prevent a significant proportion of headaches. That this effect is seen with pharmacologically unrelated classes of drugs indicates that it is likely to be due to the reduction in blood pressure per se, the only recognized action that the drugs have in common. This in turn indicates that high blood pressure is a cause of headache, but this conclusion is not supported by observational studies of blood pressure and headache. The uncertainty over whether high blood pressure causes headache does not, however, detract from the practical benefits of the use of blood pressure-lowering drugs in preventing headaches and cardiovascular disease. PMID- 16216978 TI - Enhanced activity of the myocardial Na+/H+ exchanger NHE-1 contributes to cardiac remodeling in atrial natriuretic peptide receptor-deficient mice. AB - BACKGROUND: Atrial natriuretic peptide (ANP), through its guanylyl cyclase-A (GC A) receptor, not only is critically involved in the endocrine regulation of arterial blood pressure but also locally moderates cardiomyocyte growth. The mechanisms underlying the antihypertrophic effects of ANP remain largely uncharacterized. We examined the contribution of the Na+/H+ exchanger NHE-1 to cardiac remodeling in GC-A-deficient (GC-A(-/-)) mice. METHODS AND RESULTS: Fluorometric measurements in isolated adult cardiomyocytes demonstrated that cardiac hypertrophy in GC-A(-/-) mice was associated with enhanced NHE-1 activity, alkalinization of intracellular pH, and increased Ca2+ levels. Chronic treatment of GC-A(-/-) mice with the NHE-1 inhibitor cariporide normalized cardiomyocyte pH and Ca2+ levels and regressed cardiac hypertrophy and fibrosis, despite persistent arterial hypertension. To characterize the molecular pathways driving cardiac hypertrophy in GC-A(-/-) mice, we evaluated the activity of 4 prohypertrophic signaling pathways: the mitogen-activated protein kinases (MAPK), the serine-threonine kinase Akt, calcineurin, and Ca2+/calmodulin-dependent kinase II (CaMKII). The results demonstrate that all 4 pathways were activated in GC-A(-/-) mice, but only CaMKII and Akt activity regressed during reversal of the hypertrophic phenotype by cariporide treatment. In contrast, the MAPK and calcineurin/NFAT signaling pathways remained activated during regression of hypertrophy. CONCLUSIONS: On the basis of these results, we conclude that the ANP/GC-A system moderates the cardiac growth response to pressure overload by preventing excessive activation of NHE-1 and subsequent increases in cardiomyocyte intracellular pH, Ca2+, and CaMKII as well as Akt activity. PMID- 16216979 TI - Exercise prescription and proscription for patients with coronary artery disease. PMID- 16216980 TI - Percutaneous recanalization of chronically occluded coronary arteries: a consensus document: part I. PMID- 16216981 TI - Are IA-2 and RESP18 involved in trait of blood pressure? PMID- 16216982 TI - Neurocirculatory abnormalities in Parkinson disease with orthostatic hypotension: independence from levodopa treatment. AB - Patients with Parkinson disease often have orthostatic hypotension. Neurocirculatory abnormalities underlying orthostatic hypotension might reflect levodopa treatment. Sixty-six Parkinson disease patients (36 with orthostatic hypotension, 15 off and 21 on levodopa; 30 without orthostatic hypotension) had tests of reflexive cardiovagal gain (decrease in interbeat interval per unit decrease in systolic pressure during the Valsalva maneuver; orthostatic increase in heart rate per unit decrease in pressure); reflexive sympathoneural function (decrease in pressure during the Valsalva maneuver; orthostatic increment in plasma norepinephrine); and cardiac and extracardiac noradrenergic innervation (septal myocardial 6-[18F]fluorodopamine-derived radioactivity; supine plasma norepinephrine). Severity of orthostatic hypotension did not differ between the levodopa-untreated and levodopa-treated groups with Parkinson disease and orthostatic hypotension (-52+/-6 [SEM] versus -49+/-5 mm Hg systolic). The 2 groups had similarly low reflexive cardiovagal gain (0.84+/-0.23 versus 1.33+/ 0.35 ms/mm Hg during Valsalva; 0.43+/-0.09 versus 0.27+/-0.06 bpm/mm Hg during orthostasis); and had similarly attenuated reflexive sympathoneural responses (97+/-29 versus 71+/-23 pg/mL during orthostasis; -82+/-10 versus -73+/-8 mm Hg during Valsalva). In patients off levodopa, plasma norepinephrine was lower in those with (193+/-19 pg/mL) than without (348+/-46 pg/mL) orthostatic hypotension. Low values for reflexive cardiovagal gain, sympathoneural responses, and noradrenergic innervation were strongly related to orthostatic hypotension. Parkinson disease with orthostatic hypotension features reflexive cardiovagal and sympathoneural failure and cardiac and partial extracardiac sympathetic denervation, independent of levodopa treatment. PMID- 16216983 TI - Genome-wide scan for linkage to obesity-associated hypertension in French Canadians. AB - Essential hypertension is a heterogeneous disorder that is thought to develop because of several overlapping subsets of underlying mechanisms. One such causal pathway may involve pathophysiological alterations induced by obesity. In the present study, we examined whether investigating clinically defined subtypes of hypertension, such as obesity-associated hypertension, facilitates the search for its genes. Fifty-five extended families were selected on the basis of having > or =2 siblings affected by hypertension from a geographically remote French-Canadian population. Fifteen of these families showed a high prevalence (> or =70%) of obesity. Genome-wide scan using qualitative multipoint linkage analysis (GeneHunter 2.1; marker density <10 cM) was performed in the entire set of hypertensive families and the subset with high prevalence of obesity. In the scan involving all 55 families, the most significant loci (logarithm of odds [LOD] score=2.5) were identified on chromosomes 1 (D1S1597) and 11 (D11S1999). In the scan including only the subset of families with obesity-hypertension, the most significant locus (LOD score=3.1) was found on chromosome 1 in the same region as the scan involving all families (D1S1597). Genotyping additional markers increased the significance of this locus (LOD score=3.5) and refined its position (D1S2672). Several candidate genes of obesity-hypertension are located in close proximity; these include the tumor necrosis factor receptor 2 and atrial natriuretic peptide genes. These results suggest that investigating clinically defined subtypes of hypertension, such as obesity-associated hypertension, may facilitate the search for genes of this complex disorder. PMID- 16216986 TI - Baroreflexes in hypertension: a mystery revisited. PMID- 16216984 TI - Critical role for CuZn-superoxide dismutase in preventing angiotensin II-induced endothelial dysfunction. AB - The goal of the present study was to test the hypothesis that the CuZn isoform of superoxide dismutase (CuZnSOD) protects against angiotensin II (Ang II)-induced endothelial dysfunction. Vascular responses of carotid arteries from control, CuZnSOD-deficient (CuZnSOD(+/-)), and CuZnSOD transgenic mice were examined in vitro after overnight incubation with either vehicle or Ang II (1 or 10 nmol/L). In control mice, acetylcholine produced concentration-dependent relaxation that was not affected by 1 nmol/L Ang II. In contrast, relaxation to acetylcholine in arteries from CuZnSOD+/- mice was markedly and selectively attenuated after incubation with 1 nmol/L Ang II (eg, 100 micromol/L acetylcholine produced 93+/ 6% and 44+/-15% relaxation in vehicle- and Ang II-treated arteries, respectively). A higher concentration of Ang II (10 nmol/L) selectively impaired relaxation to acetylcholine in arteries from control mice (eg, 100 micromol/L acetylcholine produced 96+/-4% and 45+/-7% relaxation in vehicle- and Ang II treated vessels, respectively). In contrast, 10 nmol/L Ang II had no effect on responses to acetylcholine in carotid arteries from CuZnSOD transgenic mice (or in control mice treated with the superoxide scavenger Tiron [1 mmol/L]). Superoxide levels in control mice were higher in aorta treated with Ang II than with vehicle and were markedly reduced in CuZnSOD transgenic mice. These findings provide the first direct evidence that CuZnSOD limits Ang II-mediated impairment of endothelial function and that loss of 1 copy of the CuZnSOD gene is sufficient to enhance Ang II-induced vascular dysfunction. PMID- 16216987 TI - Influence of prolonged baroreflex activation on arterial pressure in angiotensin hypertension. AB - Despite recent evidence indicating sustained activation of the baroreflex during chronic infusion of angiotensin II (Ang II), sinoaortic denervation does not exacerbate the severity of the hypertension. Therefore, to determine whether Ang II hypertension is relatively resistant to the blood pressure-lowering effects of the baroreflex, the carotid baroreflex was electrically activated bilaterally for 7 days in 5 dogs both in the presence and absence of a continuous infusion of Ang II (5 ng/kg per minute) producing high physiological plasma levels of the peptide. Under control conditions, basal values for mean arterial pressure (MAP) and plasma norepinephrine concentration (NE) were 93+/-1 mm Hg and 99+/-25 pg/mL, respectively. By day 7 of baroreflex activation, MAP and NE were reduced to 72+/ 4 mm Hg (-21+/-3 mm Hg) and 56+/-15 pg/mL, respectively, but PRA was unchanged (control=0.41+/-0.06 ng ANG I/mL per hour). All values returned to basal levels by the end of a 7-day recovery period. After 7 days of Ang II infusion, MAP increased from 93+/-3 to 129+/-3 mm Hg, whereas NE fell from 117+/-15 to 86+/-23 pg/mL. During the next 7 days of baroreflex activation/Ang II infusion, further reductions in NE were not statistically significant, and on the final day of baroreflex activation, the reduction in MAP was only 5+/-1 mm Hg, compared with 21+/-3 mm Hg in the control normotensive state. These findings indicate that long term baroreflex-mediated reductions in arterial pressure are markedly diminished, but not totally eliminated, in the presence of hypertension produced by chronic infusion of Ang II. PMID- 16216988 TI - Activation of vascular BK channel by tempol in DOCA-salt hypertensive rats. AB - Large-conductance Ca2+-activated potassium (BK) channels modulate vascular smooth muscle tone. Tempol, a superoxide dismutase (SOD) mimetic, lowers blood pressure and inhibits sympathetic nerve activity in normotensive and hypertensive rats. In the present study, we tested the hypotheses depressor responses caused by tempol are partly mediated by vasodilation. It was found that tempol, but not tiron (a superoxide scavenger), dose-dependently relaxed mesenteric arteries (MA) in anesthetized sham and deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Tempol also reduced perfusion pressure in isolated, norepinephrine (NE) preconstricted MA from sham and DOCA-salt hypertensive rats. Maximal responses in DOCA-salt rats were twice as large as those in sham rats. The vasodilation caused by tempol was blocked by iberiotoxin (IBTX, BK channel antagonist, 0.1 micromol/L) and tetraethylammonium chloride (TEA) (1 mmol/L). Tempol did not relax KCl preconstricted arteries in sham or DOCA-salt rats, and Nomega-nitro-L arginine methyl ester (L-NAME), apamin, or glibenclamide did not alter tempol induced vasodilation. IBTX constricted MA and this response was larger in DOCA salt compared with sham rats. Western blots and immunohistochemical analysis revealed increased expression of BK channel alpha subunit protein in DOCA-salt arteries compared with sham arteries. Whole-cell patch clamp studies revealed that tempol enhanced BK channel currents in HEK-293 cells transiently transfected with mslo, the murine BK channel a subunit. These currents were blocked by IBTX. The data indicate that tempol activates BK channels and this effect contributes to depressor responses caused by tempol. Upregulation of the BK channel alpha subunit contributes to the enhanced depressor response caused by tempol in DOCA salt hypertension. PMID- 16216989 TI - Sympathectomy or doxazosin, but not propranolol, blunt myocardial interstitial fibrosis in pressure-overload hypertrophy. AB - The adaptive changes that develop in the pressure-overloaded left ventricular (LV) myocardium include cardiomyocyte hypertrophy and interstitial fibrosis. Although the former is known to depend to a sizeable extent on sympathetic (over)activity, little information exists whether the same applies to the latter, ie, whether excess catecholamine exposure contributes to the imbalance between collagen deposition by fibroblasts and degradation by matrix metalloproteases (MMPs), eventually leading to LV collagen accumulation. Sprague-Dawley rats were subjected to abdominal aortic banding (B) or sham operation (S) and treated with beta-blockade (Bb, oral propranolol, 40 mg/kg per day), chemical sympathectomy (Sx, 6-hydroxydopamine, 150 mg/kg intraperitoneal twice per week) or vehicle (Vh). Ten weeks later, systolic blood pressure, LV weight, collagen abundance (computer-aided histology), zymographic matrix metalloproteinase (MMP)-2 activity and its specific tissue inhibitor concentration (TIMP-2) were measured. Both sympathectomy and beta-blockade failed to attenuate the banding-induced blood pressure elevation but significantly attenuated the attendant LV hypertrophy. As expected, pressure-overload hypertrophy was associated with interstitial fibrosis (collagen: 4.37+/-1.23% BVh versus 1.23+/-0.44% SVh, P<0.05), which was abolished by sympathectomy (2.55+/-1.31%, P=not significant versus SSx) but left unchanged by beta-blockade (4.11+/-1.23%, P<0.05 versus both SBb and BSx). beta-blockade, but not sympathectomy, was also associated with an increased TIMP-2/MMP-2 ratio (P<0.05), indicating reduced interstitial collagenolytic activity. In separate groups of banded and sham-operated rats, treatment with the alpha-receptor blocker doxazosin (10 mg/kg per day) displayed similar antifibrotic and biochemical effects as sympathectomy. Thus in the course of experimental pressure overload, the sympathetic nervous system plays a major pro-fibrotic role, which is mediated via alpha-adrenergic but not beta-adrenergic receptors. PMID- 16216990 TI - Gender-specific alteration of adrenergic responses in small femoral arteries from estrogen receptor-beta knockout mice. AB - Estrogen receptor-beta knockout mice become hypertensive as they age, and males have a higher blood pressure than females. We hypothesized that the absence of estrogen receptor-beta may contribute to development of cardiovascular dysfunction by modification of adrenergic responsiveness in the peripheral vasculature. Small femoral arteries (internal diameter <200 microm) were isolated from estrogen receptor-beta knockout and wild-type mice and mounted on a wire myograph. Concentration-response curves to phenylephrine and norepinephrine were compared and the contribution of adrenoceptor subtypes established using specific agonists and antagonists. The involvement of endothelial factors in the modulation of resting tone was also investigated and immunohistochemical analysis used to confirm the presence or absence of estrogen receptor expression. Compared with wild type, arteries from estrogen receptor-beta knockout male, but not female, mice demonstrated gender-specific enhancement of the response to phenylephrine (alpha1-adrenoceptor agonist), which was accompanied by elevated basal tension attributable to endothelial factors. Contractile responses to the mixed adrenoceptor agonist norepinephrine did not differ significantly between estrogen receptor-beta knockout and wild type; however, beta-adrenoceptor inhibition unmasked an enhanced underlying alpha1-adrenoceptor responsiveness in estrogen receptor-beta knockout males. beta-adrenoceptor-mediated dilatation was also enhanced in estrogen receptor-beta knockout versus wild-type males. We suggest that estrogen receptor-beta modifies the adrenergic control of small artery tone in males but not in females. PMID- 16216991 TI - Arterial stiffness is related to systemic inflammation in essential hypertension. AB - The acute phase-reactant high-sensitivity C-reactive protein, a marker of vascular inflammation and an atherosclerotic risk factor, is related to arterial stiffness in healthy subjects and in systemic vasculitis. To explore the relationship between markers of inflammation, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and high-sensitivity C-reactive protein with arterial stiffness, we studied untreated patients (n=78; 56% male; 47+/-1 years of age; mean+/-SEM) with essential hypertension. After overnight fast, augmentation index and pulse wave velocity were assessed noninvasively and related to plasma levels of inflammatory markers measured by ELISA. Pulse wave velocity was significantly related to plasma high-sensitivity C-reactive protein (r=0.31; P<0.001), TNF-alpha, (r=0.30; P<0.001) and IL-6 (r=0.21; P<0.05). There was also a relationship between heart rate-corrected augmentation index to high sensitivity C-reactive protein (r=0.37; P<0.001), IL-6 (r=0.24; P<0.05), and TNF alpha (r=0.19, P=0.06). High-sensitivity C-reactive protein was an independent predictor of pulse wave velocity and augmentation index in a multiple stepwise regression model. High-sensitivity C-reactive protein, a marker of systemic inflammation, is independently related to pulse wave velocity, a marker of aortic stiffness, and augmentation index, a manifestation of wave reflection, in essential hypertension. PMID- 16216992 TI - Does the angiotensin-converting enzyme (ACE)/ACE2 balance contribute to the fate of angiotensin peptides in programmed hypertension? PMID- 16216993 TI - Eplerenone antagonizes atherosclerosis, but what is the agonist? PMID- 16216994 TI - C-reactive protein, heart disease risk, and the popular media. PMID- 16216995 TI - High attributable risk of elevated C-reactive protein level to conventional coronary heart disease risk factors: the Third National Health and Nutrition Examination Survey. AB - BACKGROUND: C-reactive protein (CRP), a marker of systemic inflammation, is predictive of coronary heart disease (CHD) events. However, the extent to which high CRP levels (>3 mg/L) may be attributable to high cholesterol levels and other CHD risk factors has not been well defined. METHODS: The prevalence of high CRP levels in the third National Health and Nutrition Examination Survey (n = 15 341) was studied using CHD risk-factor cut points designated as abnormal (total cholesterol values, >or=240 mg/dL [>or=6.22 mmol/L]; fasting blood glucose levels, >or=126 mg/dL [>or=6.99 mmol/L]; blood pressure, >or=140/90 mm Hg; body mass index [BMI], >or=30 kg/m(2); high-density lipoprotein cholesterol values, <40 mg/dL [<1.04 mmol/L] for men and <50 mg/dL [<1.30 mmol/L] for women; triglyceride levels, >or=200 mg/dL [>or=2.26 mmol/L]; current smoking status) or borderline (total cholesterol values, 200-239 mg/dL [5.18-6.19 mmol/L]; fasting blood glucose levels, 100-125 mg/dL [5.55-6.94 mmol/L]; blood pressure, 120 139/80-89 mm Hg; BMI, 25.0-29.9 kg/m(2), and triglyceride values 150-199 mg/dL [1.70-2.25 mmol/L], former smoking status), or normal. RESULTS: Weighted multiple logistic regression analysis demonstrated that high CRP level was significantly more common with obesity (odds ratio [OR], 3.78; 95% confidence interval [CI], 3.28-4.35]), overweight (OR, 1.88; 95% CI, 1.62-2.18), and diabetes (OR, 1.91; 95% CI, 1.54-2.38) and that high CRP level was rare in the absence of any borderline or abnormal CHD risk factor in men (4.4%) and women (10.3%). Overall, the risk of elevated CRP level attributable to the presence of any abnormal or borderline CHD risk factor was 78% in men and 67% women. CONCLUSIONS: These data suggest that elevated CRP levels in the general population are in large measure attributable to traditional CHD risk factors. Moreover, CRP level elevation is rare in the absence of borderline or abnormal risk factors. As such, CRP measurements may have limited clinical utility as a screening tool beyond other known CHD risk factors. PMID- 16216996 TI - The complexity and cost of drug regimens of older patients hospitalized with heart failure in the United States, 1998-2001. AB - BACKGROUND: Polypharmacy-the concurrent prescription of multiple medications-is a salient consideration in the care of older patients with heart failure. Little is known, however, about the complexity and financial burden of medical therapy in this population. METHODS: This is a study of the chronic medications prescribed at hospital discharge to patients 65 years or older hospitalized for heart failure in 2 cohorts separated by 27 months (April 1998-March 1999, n = 31 602; July 2000-June 2001, n = 30,774). Three utilization measures were assessed: the number of drugs, the estimated number of doses per day, and the estimated annual costs using the same cost standard (2003 average wholesale prices) for both samples. Utilization associated with population characteristics and between time frames was assessed in multivariable models. RESULTS: In 1998-1999, the mean number of drugs was 6.8, representing 10.1 doses daily at a cost of 3142 dollars/y, increasing to 7.5 drugs, 11.1 doses daily and 3823 dollars/y in 2000 2001 (P<.001 for all comparisons). After adjustment, the number of drugs increased by 12% and costs by 24% between samples. Factors associated with greater complexity and cost included diabetes (1.6 additional drugs and 1094 dollars/y additional cost), prior revascularization (1.3 drugs, 1154 dollars/y), and chronic lung disease (1.2 drugs, 814 dollars/y). Younger age and white race were also associated with more drugs and higher costs. CONCLUSIONS: The drug treatment of older patients with heart failure is characterized by rapidly increasing complexity and cost. Efforts should be directed toward optimizing the complex drug regimens of elderly patients with heart failure and multiple comorbidities. PMID- 16216997 TI - The influence of health status, age, and race on screening mammography in elderly women. AB - BACKGROUND: Screening mammography is controversial for elderly women because of an absence of efficacy data. Decisions to screen are based on individualized assessment of risks and benefits. Our objective was to determine how screening mammography varies by age and race when adjusted for propensity to die. METHODS: In a retrospective cohort study, rates of screening mammogram performed in 2000 2001 based on claims, adjusted for propensity to die in 2000, were determined for a nationally representative 5% random sample of female fee-for-service Medicare beneficiaries 65 years and older in (N = 722,310). RESULTS: The overall rate of screening was 39%. When stratified into quintiles by propensity to die, 2-year rates ranged from 61% in the lowest-risk group to 5% in the highest-risk group. In analyses stratified by age and adjusted for propensity to die, 42% of women aged 65 to 69 years were screened, declining to 26% of women 85 years and older (P<.001). Adjusted screening rates for white women, black women, and women of other races were 40%, 30%, and 25%, respectively (P<.001). Thus, among women with similar health status, the youngest women were 1.61 times more likely to be screened compared with the oldest; compared with black women and women of other races, white women were 1.38 and 1.60 times, respectively, more likely to be screened. CONCLUSIONS: Decisions to screen for breast cancer are related not only to health status but also to age and race. Underuse and overuse of screening mammography likely occurs owing to age- and race-associated decision making. Assessment of life expectancy may more accurately identify women who could benefit from screening. PMID- 16216998 TI - Reduced screening mammography among women with diabetes. AB - BACKGROUND: Despite regular health care, preventive health issues may be neglected in patients with chronic diseases such as diabetes. Case-control studies in the United States have shown lower mammogram rates in women with diabetes; however, it is not known whether the presence of diabetes mellitus affects mammography use in a Canadian setting, where there is universal access to health care. METHODS: Using health databases in Ontario from April 1, 1999, to March 31, 2002, this retrospective cohort study observed women aged 50 to 67 years, who were free of breast cancer, until their first mammogram in a 2-year period. Mammogram rates were compared between women who had had diabetes for a minimum of 2 years (n = 69 168) and women without diabetes (n = 663 519). RESULTS: Compared with women without diabetes, diabetic patients were older, had more physician visits, were more often from a lower-income neighborhood, and, in those 65 years or older, were less likely to be taking estrogen. The odds ratio of having a mammogram during the 2-year period was 0.68 (95% confidence interval, 0.67-0.70; P<.001) for women with diabetes, and adjustment for age and other covariates did not modify this effect. CONCLUSIONS: Women with diabetes were significantly less likely to have had a mammogram during a 2-year period than were women without diabetes, despite more health care visits. These results suggest that, because of the complexity involved in diabetes care, routine preventive care such as cancer screening is often neglected. These findings highlight the need for better organization of primary care for patients with chronic diseases. PMID- 16216999 TI - Racial disparities in hypertension prevalence, awareness, and management. AB - BACKGROUND: Effectively reducing cardiovascular disease disparities requires identifying and reducing disparities in risk factors. Improved understanding of hypertension disparities is critical. METHODS: Cross-sectional analysis of nationally representative samples of black and white adults 20 years and older who participated in the National Health and Nutrition Examination Survey (NHANES) 1999-2002 (white, n = 4624; black, n = 1837) and NHANES III conducted in 1988 1994 (white, n = 7121; black, n = 4709). We examined differences in hypertension prevalence, awareness, treatment, and blood pressure (BP) control among both treated and prevalent cases across the 2 periods. RESULTS: Hypertension prevalence increased significantly from 35.8% to 41.4% among blacks and from 24.3% to 28.1% among whites and remains significantly higher among blacks. Awareness is higher among blacks (77.7% vs 70.4%; P<.001), as is treatment (68.2% vs 60.4%; P<.001). These results are driven by higher rates in black women. Blood pressure control rates among those treated have increased in both races, primarily as a result of increased BP control in black and white men (27.3% and 44.7%, respectively; Por=30) at different levels of income. RESULTS: Over the course of 3 decades, obesity has increased at all levels of income. Moreover, it is typically not the poor who have experienced the largest gains. For example, among black women, the absolute increase in obesity is 27.0% (1.05% per year) for those at middle incomes, but only 14.5% (0.54% per year) for the poor. Among black men, the increase in obesity is 21.1% (0.77% per year) for those at the highest level of income, but only 4.5% (0.06% per year) for the near poor and 5.4% (0.50% per year) for the poor. Furthermore, all race-sex groups show income differentials on body mass index, but patterns show substantial variation between groups and consistency and change within groups over time. For example, white women consistently show a strong inverse gradient, while a positive gradient emerges in later waves for black and Mexican American men. CONCLUSION: The persistence and emergence of income gradients suggests that disparities in weight status are only partially attributable to poverty and that efforts aimed at reducing disparities need to consider a much broader array of contributing factors. PMID- 16217003 TI - Disparities despite coverage: gaps in colorectal cancer screening among Medicare beneficiaries. AB - BACKGROUND: Despite its effectiveness in reducing mortality, colorectal cancer (CRC) screening rates are low, especially among low-income and minority groups; however, physician recommendation can increase screening rates. METHODS: We performed a multilevel analysis of the Medicare Current Beneficiary Survey data linked to Medicare claims and the Area Resource File to identify determinants of racial and socioeconomic disparities in CRC screening among 9985 Medicare Parts A and B beneficiaries with a usual physician. Recent CRC screening was defined as receipt of either a home fecal occult blood test, flexible sigmoidoscopy, or colonoscopy at recommended intervals. RESULTS: Unadjusted rates of screening were 48% for white and 39% for black beneficiaries (P<.001). Racial differences in CRC screening receipt were eliminated after adjustment for socioeconomic status as measured by income and education. Socioeconomic status disparities decreased but remained significant after adjustment for personal and health system factors. Awareness of CRC (adjusted odds ratio, 2.76; 95% confidence interval, 2.29-3.33) and having a primary care generalist (vs another specialist) as one's usual physician (adjusted odds ratio, 1.31; 95% confidence interval, 1.12-1.53) were associated with higher odds of screening, controlling for other factors. The odds of screening were also higher among those whose usual physician was rated more highly on information-giving skills. CONCLUSIONS: Racial differences in CRC screening rates among Medicare beneficiaries with a usual physician are explained by differences in socioeconomic status. Beneficiaries with a primary care generalist as their usual physician had higher rates of CRC screening receipt. Increased efforts to make Medicare beneficiaries aware of the benefits of CRC screening may capitalize on the associations found in this study between CRC knowledge, physician information giving, and timely screening. PMID- 16217004 TI - Availability of physician services in Florida, revisited: the effect of the professional liability insurance market on access to health care. AB - BACKGROUND: Access to care remains a key part of improving health care outcomes in the United States. Recent reports have suggested that the number of physicians able to meet the demands for access to care may be decreasing. METHODS: We surveyed physicians practicing in rural and urban/suburban areas of Florida in 2004 to determine whether changes were occurring in health care service delivery. Secondary outcomes assessed included changes in professional liability insurance and their possible effects on changes in service delivery. RESULTS: Overall, 727 (54.4%) of responding physicians stated that the delivery of services had been decreased or eliminated in the previous year. The most commonly eliminated services were nursing home coverage (42.1%), vaginal deliveries (29.1%), cesarean deliveries (26.0%), emergency department coverage (22.8%), and mental health services (21.2%). Surgical specialists (70.2%) and general surgeons (68.5%) were the groups with the highest number of decreased or eliminated services, but this trend was broad, with 63.6% of obstetrician/gynecologists and 60.2% of family medicine physicians also decreasing or eliminating services. Decreases in services seem to be related to changes in professional liability insurance premiums when assessed by both percentage of change and total premium increases for physicians. Rural and urban/suburban physicians did not differ significantly in these assessments. CONCLUSION: The findings suggest that physicians across Florida have continued to decrease or eliminate important health services and that these decreases seem to be related to the difficulty of finding or paying for professional liability insurance. PMID- 16217005 TI - The relation between pulse pressure and cardiovascular mortality in 12,763 middle aged men from various parts of the world: a 25-year follow-up of the seven countries study. AB - BACKGROUND: Hypertension is a dominant characteristic in the prediction of cardiovascular diseases (CVDs). We aimed to evaluate the association of blood pressure measurements with CVD mortality among different populations of the world. METHODS: A total of 12 763 men, aged 40 to 59 years, from 7 countries (United States, Japan, Italy, Greece, former Yugoslavia, Finland, and the Netherlands) were surveyed from 1958 to 1964. Follow-up for vital status and causes of death was carried out over 25 years. RESULTS: All baseline blood pressure measurements were the best predictors of CVD mortality, compared with age, physical activity, total serum cholesterol level, body mass index or height, and smoking. Moreover, pulse pressure and diastolic and systolic blood pressures were the best predictors for CVD death, followed by mean and mid blood pressures. The age-adjusted hazard ratio per 10-mm Hg increase in pulse pressure varied among cohorts from 1.19 in the United States (P = .04) to 1.29 in southern Europe (P = .01). Differences among cohorts were not significant. In the pooled cohorts, pulse pressure measurements were also a significant predictor for coronary heart disease (hazard ratio per 10-mm Hg increase, 1.15; P = .04) as well as stroke death (hazard ratio per 10-mm Hg increase, 1.32; P = .01). CONCLUSIONS: Pulse pressure followed by diastolic and systolic blood pressures were the best predictors for CVD mortality among other blood pressures, as well as age, physical activity, total serum cholesterol level, anthropometric indexes, and smoking habits. No significant differences were observed among the different populations studied. PMID- 16217007 TI - Continued decline in blood lead levels among adults in the United States: the National Health and Nutrition Examination Surveys. AB - BACKGROUND: Declines in blood lead levels between 1976 and 1991 among US adults have been previously reported. More recent trends in blood lead levels and the association of lower blood lead levels with chronic disease have not been reported. METHODS: Data from 2 nationally representative cross-sectional surveys, the Third National Health and Nutrition Examination Survey conducted in 1988-1994 (n = 16,609) and the National Health and Nutrition Examination Survey conducted in 1999-2002 (n = 9961) were analyzed. RESULTS: The geometric mean blood lead level declined 41% from 2.76 microg/dL (0.13 micromol/L) in 1988-1994 to 1.64 microg/dL (0.08 micromol/L) in 1999-2002. The percentage of adults with blood lead levels of 10 microg/dL (0.48 micromol/L) or higher declined from 3.3% in 1988-1994 to 0.7% in 1999-2002 (P<.001). In 1999-2002, the multivariable-adjusted odds ratio of having a blood lead level of 10 microg/dL (0.48 micromol/L) or higher was 2.91 (95% confidence interval [CI], 1.74-4.84) and 3.26 (1.83-5.81) for non-Hispanic blacks and Mexican Americans, respectively, compared with non Hispanic whites. After multivariable adjustment, persons in the highest quartile (>or=2.47 microg/dL [>or=0.12 micromol/L]) compared with those in the lowest quartile (<1.06 microg/dL [<0.05 micromol/L]) of blood lead levels were 2.72 (95% CI, 1.47-5.04) and 1.92 (95% CI, 1.02-3.61) times more likely to have chronic kidney disease and peripheral arterial disease, respectively. In addition, higher blood lead levels were associated with a higher multivariable-adjusted odds ratio of hypertension among non-Hispanic blacks and Mexican Americans. CONCLUSIONS: Blood lead levels continue to decline among US adults, but racial and ethnic disparities persist. Higher blood lead levels remain associated with a higher burden of chronic kidney and peripheral arterial diseases among the overall population and with hypertension among non-Hispanic blacks and Mexican Americans. PMID- 16217006 TI - Lipoprotein peroxidation and mobility limitation: results from the Health, Aging, and Body Composition Study. AB - BACKGROUND: Oxidative damage plays an important role in leading to major health related events. The aim of this study was to assess the predictive value of a lipoprotein peroxidation marker, oxidized low-density lipoprotein (oxLDL) for incident mobility limitation (ML). METHODS: Data are from 2985 well-functioning elders enrolled in the Health ABC study (median follow-up, 4.1 years). All oxLDL levels were measured at the baseline assessment. The oxLDL/LDL cholesterol (LDL C) ratio (log value) was used as a measure of lipoprotein peroxidation. Mobility limitation was defined by 2 consecutive semiannual reports of any difficulty either walking 1/4 mile or climbing up 10 steps without resting. Severe ML was defined by 2 consecutive reports of great difficulty or inability to do the same tasks. Cox proportional hazards models were performed to assess hazard ratios (HRs) and 95% confidence intervals (CIs). RESULTS: The mean (SD) age of the sample was 74.2 (2.9) years. After adjustment for potential confounders (sociodemographic factors, smoking, physical activity, body mass index, clinical conditions, biological markers, and medications), the relationship between the oxLDL/LDL-C ratio and disability events was statistically significant (per log unit difference in the oxLDL/LDL-C ratio) (for ML: HR, 1.22; 95% CI, 1.06-1.41; for severe ML: HR, 1.43; 95% CI, 1.15-1.79). Consistent results were found when interleukin 6 level was included as a covariate in the adjusted models (ML: HR, 1.13; 95% CI, 0.98-1.31; severe ML: HR, 1.31; 95% CI, 1.05-1.64). No significant sex, race, interleukin 6 level, or clinical conditions interaction was found with the oxLDL/LDL-C ratio and mobility disability. CONCLUSIONS: Lipoprotein peroxidation predicts the onset of ML in older persons. The oxLDL predictive value for ML is partly explained by interleukin 6 levels. PMID- 16217008 TI - Left ventricular assessment in myocardial infarction: the VALIANT registry. AB - BACKGROUND: How often echocardiography and cardiac catheterization are used to evaluate left ventricular (LV) function in patients with myocardial infarction (MI) and how they are associated with quality of care is unknown. METHODS: Patients with MI in the Valsartan in Acute Myocardial Infarction (VALIANT) registry were divided into those with (n = 1423) and without (n = 3968) heart failure (HF), and the use of either echocardiography or cardiac catheterization for LV assessment in each group was compared along with associated baseline characteristics. We evaluated the association between LV assessment and discharge medications. Using a multivariable model with a propensity analysis, we evaluated the association of LV assessment with in-hospital outcomes. RESULTS: Of the patients with HF, 322 (22.6%) had no LV assessment. Patients with HF with LV assessment were discharged more frequently under treatment with aspirin (81.3% vs 70.0%; P<.001), beta-blockers (65.6% vs 56.4%; P = .008), clopidogrel (30.4% vs 14.0%; P<.001), and statins (45.9% vs 34.2%; P<.001). Patients without HF who underwent LV assessment were discharged more frequently under treatment with an angiotensin-converting enzyme inhibitor (53.8% vs 41.5%; P<.001). After adjustment for regional use, other covariates, and revascularization, LV assessment was associated with lower in-hospital mortality in patients with HF (adjusted odds ratio [OR], 0.45; P<.001) and in patients without HF (adjusted OR, 0.30; P<.001). After excluding deaths during the first 2 days, LV assessment remained associated with lower mortality in patients with HF (adjusted OR, 0.59; P = .03) and in patients without HF (adjusted OR, 0.41; P<.001). CONCLUSION: Left ventricular assessment was frequently not performed during the in-hospital stay of patients with acute MI, including those with clinical HF, and its use was associated with better quality of care. PMID- 16217009 TI - On the other side of the fence. PMID- 16217010 TI - Low-carbohydrate diet study has serious flaws. PMID- 16217011 TI - The evolutionary origin of a complex scrambled gene. AB - Some species of ciliates undergo massive DNA elimination and genome rearrangement to construct gene-sized "chromosomes" in their somatic nucleus. An example is the extensively scrambled DNA polymerase alpha gene that is broken into 48 pieces and distributed over two unlinked loci in Stylonychia. To understand the emergence of this complex phenomenon during evolution, we examined DNA polymerase alpha genes in several earlier diverging species, representing evolutionary intermediates. Mapping these data onto an evolutionary tree suggests that this gene became extensively fragmented and scrambled over evolutionary time through a series of steps, each leading to greater complexity. Our results also suggest a possible mechanism for intron loss by deletion of intron sequences as DNA during development of the somatic nucleus. PMID- 16217012 TI - Interface connections of a transmembrane voltage sensor. AB - Voltage-sensitive ion channels open and close in response to changes in transmembrane (TM) potential caused by the motion of the S4 voltage sensors. These sensors are alpha-helices that include four or more positively charged amino acids, most commonly arginine. The so-called paddle model, based on the high-resolution structure of the KvAP K+ channel [Jiang, et al. (2003) Nature 423, 33-41], posits that the S4 sensors move within the membrane bilayer in response to TM voltage changes. Direct exposure of S4 sensors to lipid is contrary to the classical expectation that the dielectric contrast between the membrane hydrocarbon core and water presents an insurmountable energetic penalty to burial of electric charges. Nevertheless, recent experiments have shown that a helix with the sequence of KvAP S4 can be inserted across the endoplasmic reticulum membrane. To reconcile this result with the classical energetics argument, we have carried out a molecular dynamics simulation of an isolated TM S4 helix in a lipid bilayer. The simulation reveals a stabilizing hydrogen-bonded network of water and lipid phosphates around the arginines that reduces the effective thickness of the bilayer hydrocarbon core to approximately 10 A in the vicinity of the helix. It suggests that bilayer phospholipids can adapt locally to strongly perturbing protein elements, causing the phospholipids to become a structural extension of the protein. PMID- 16217013 TI - Heterogeneous nuclear ribonucleoprotein C1/C2, MeCP1, and SWI/SNF form a chromatin remodeling complex at the beta-globin locus control region. AB - Locus control regions (LCRs) are regulatory DNA sequences that are situated many kilobases away from their cognate promoters. LCRs protect transgenes from position effect variegation and heterochromatinization and also promote copy number dependence of the levels of transgene expression. In this work, we describe the biochemical purification of a previously undescribed LCR-associated remodeling complex (LARC) that consists of heterogeneous nuclear ribonucleoprotein C1/C2, nucleosome remodeling SWI/SNF, and nucleosome remodeling and deacetylating (NuRD)/MeCP1 as a single homogeneous complex. LARC binds to the hypersensitive 2 (HS2)-Maf recognition element (MARE) DNA in a sequence-specific manner and remodels nucleosomes. Heterogeneous nuclear ribonucleoprotein C1/C2, previously known as a general RNA binding protein, provides a sequence-specific DNA recognition element for LARC, and the LARC DNA-recognition sequence is essential for the enhancement of transcription by HS2. Independently of the initiation of transcription, LARC becomes associated with beta-like globin promoters. PMID- 16217015 TI - A spider that feeds indirectly on vertebrate blood by choosing female mosquitoes as prey. AB - Spiders do not feed directly on vertebrate blood, but a small East African jumping spider (Salticidae), Evarcha culicivora, feeds indirectly on vertebrate blood by choosing as preferred prey female mosquitoes that have had recent blood meals. Experiments show that this spider can identify its preferred prey by sight alone and by odor alone. When presented with two types of size-matched motionless lures, E. culicivora consistently chose blood-fed female mosquitoes in preference to nonmosquito prey, male mosquitoes, and sugar-fed female mosquitoes (i.e., females that had not been feeding on blood). When the choice was between mosquitoes of different sizes (both blood- or both sugar-fed), small juveniles chose the smaller prey, whereas adults and larger juveniles chose the larger prey. However, preference for blood took precedence over preference for size (i.e., to get a blood meal, small individuals took prey that were larger than the preferred size, and larger individuals took prey that were smaller than the preferred size). When presented with odor from two prey types, E. culicivora approached the odor from blood-fed female mosquitoes significantly more often the odor of the prey that were not carrying blood. PMID- 16217014 TI - Ribosomal S6 kinase 2 interacts with and phosphorylates PDZ domain-containing proteins and regulates AMPA receptor transmission. AB - Extracellular signal-regulated kinase (ERK) signaling is important for neuronal synaptic plasticity. We report here that the protein kinase ribosomal S6 kinase (RSK)2, a downstream target of ERK, uses a C-terminal motif to bind several PDZ domain proteins in heterologous systems and in vivo. Different RSK isoforms display distinct specificities in their interactions with PDZ domain proteins. Mutation of the RSK2 PDZ ligand does not inhibit RSK2 activation in intact cells or phosphorylation of peptide substrates by RSK2 in vitro but greatly reduces RSK2 phosphorylation of PDZ domain proteins of the Shank family in heterologous cells. In primary neurons, NMDA receptor (NMDA-R) activation leads to ERK and RSK2 activation and RSK-dependent phosphorylation of transfected Shank3. RSK2-PDZ domain interactions are functionally important for synaptic transmission because neurons expressing kinase-dead RSK2 display a dramatic reduction in frequency of AMPA-type glutamate receptor-mediated miniature excitatory postsynaptic currents, an effect dependent on the PDZ ligand. These results suggest that binding of RSK2 to PDZ domain proteins and phosphorylation of these proteins or their binding partners regulates excitatory synaptic transmission. PMID- 16217016 TI - Rundown of GABA type A receptors is a dysfunction associated with human drug resistant mesial temporal lobe epilepsy. AB - Pharmacotherapeutic strategies have been difficult to develop for several forms of temporal lobe epilepsy, which are consequently treated by surgical resection. To examine this problem, we have studied the properties of transmitter receptors of tissues removed during surgical treatment. We find that when cell membranes, isolated from the temporal neocortex of patients afflicted with drug-resistant mesial temporal lobe epilepsy (TLE), are injected into frog oocytes they acquire GABA type A receptors (GABA(A)-receptors) that display a marked rundown during repetitive applications of GABA. In contrast, GABA(A)-receptor function is stable in oocytes injected with cell membranes isolated from the temporal lobe of TLE patients afflicted with neoplastic, dysgenetic, traumatic, or ischemic temporal lesions (lesional TLE, LTLE). Use-dependent GABA(A)-receptor rundown is also found in the pyramidal neurons of TLE neocortical slices and is antagonized by BDNF. Pyramidal neurons in cortical slices of a traumatic LTLE patient did not show GABA(A)-receptor rundown. However, the apparent affinity of GABA(A)-receptor in oocytes microtransplanted with membranes from all of the epileptic patients studied was smaller than the affinity of receptors transplanted from the nonepileptic brain. We conclude that the use-dependent rundown of neocortical GABA(A)-receptor represents a TLE-specific dysfunction, whereas the reduced affinity may be a general feature of brains of both TLE and LTLE patients, and we speculate that our findings may help to develop new treatments for TLE and LTLE. PMID- 16217017 TI - Dynamic effects of antibody-dependent enhancement on the fitness of viruses. AB - Antibody-dependent enhancement (ADE), a phenomenon in which viral replication is increased rather than decreased by immune sera, has been observed in vitro for a large number of viruses of public health importance, including flaviviruses, coronaviruses, and retroviruses. The most striking in vivo example of ADE in humans is dengue hemorrhagic fever, a disease in which ADE is thought to increase the severity of clinical manifestations of dengue virus infection by increasing virus replication. We examine the epidemiological impact of ADE on the prevalence and persistence of viral serotypes. Using a dynamical system model of n cocirculating dengue serotypes, we find that ADE may provide a competitive advantage to those serotypes that undergo enhancement compared with those that do not, and that this advantage increases with increasing numbers of cocirculating serotypes. Paradoxically, there are limits to the selective advantage provided by increasing levels of ADE, because greater levels of enhancement induce large amplitude oscillations in incidence of all dengue virus infections, threatening the persistence of both the enhanced and nonenhanced serotypes. Although the models presented here are specifically designed for dengue, our results are applicable to any epidemiological system in which partial immunity increases pathogen replication rates. Our results suggest that enhancement is most advantageous in settings where multiple serotypes circulate and where a large host population is available to support pathogen persistence during the deep troughs of ADE-induced large amplitude oscillations of virus replication. PMID- 16217018 TI - Making ATP. AB - We present a mesoscopic model for ATP synthesis by F(1)F(o) ATPase. The model combines the existing experimental knowledge of the F(1) enzyme into a consistent mathematical model that illuminates how the stages in synthesis are related to the protein structure. For example, the model illuminates how specific interactions between the gamma, epsilon, and alpha(3)beta(3) subunits couple the F(o) motor to events at the catalytic sites. The model also elucidates the origin of ADP inhibition of F(1) in its hydrolysis mode. The methodology we develop for constructing the structure-based model should prove useful in modeling other protein motors. PMID- 16217020 TI - Partition congruences and the Andrews-Garvan-Dyson crank. AB - In 1944, Freeman Dyson conjectured the existence of a "crank" function for partitions that would provide a combinatorial proof of Ramanujan's congruence modulo 11. Forty years later, Andrews and Garvan successfully found such a function and proved the celebrated result that the crank simultaneously "explains" the three Ramanujan congruences modulo 5, 7, and 11. This note announces the proof of a conjecture of Ono, which essentially asserts that the elusive crank satisfies exactly the same types of general congruences as the partition function. PMID- 16217019 TI - Inhibition of vagally mediated immune-to-brain signaling by vanadyl sulfate speeds recovery from sickness. AB - To the ill patient with diabetes, the behavioral symptoms of sickness such as fatigue and apathy are debilitating and can prevent recuperation. Here we report that peripherally administered insulin-like growth factor 1 (IGF-1) attenuates LPS-dependent depression of social exploration (sickness) in nondiabetic (db/+) but not in diabetic (db/db) mice. We show that the insulin/IGF-1 mimetic vanadyl sulfate (VS) is effective at augmenting recovery from sickness in both db/+ and db/db mice. Specifically, peak illness was reached at 2 h for both VS and control animals injected with LPS, and VS mice recovered 50% faster than non-VS-treated animals. Examination of the mechanism of VS action in db/+ mice showed that VS paradoxically augmented peritoneal macrophage responsivity to LPS, increasing both peritoneal and ex vivo macrophage production of IL-1beta and IL-6 but not TNF-alpha. The effects of VS in promoting recovery from sickness were not restricted to LPS, because they were also observed after direct administration of IL-1beta. To explore the possibility that VS impairs immune-to-brain communication via vagal afferents, the vagally mediated satiety-inducing effects of cholecystokinin 8 were tested in db/+ mice. Cholecystokinin decreased food intake in saline-injected mice but not in VS-treated mice. VS also inhibited LPS dependent up-regulation of IL-1beta and IL-6 mRNA in the brain, while increasing by 50% the cerebral expression of transcripts of the specific antagonist of IL-1 receptors IL-1RA and IL-1R2. Taken together, these data indicate that VS improves recovery from LPS-induced sickness by blocking vagally mediated immune-to-brain signaling and by up-regulating brain expression of IL-1beta antagonists. PMID- 16217021 TI - Visible and UV coherent Raman spectroscopy of dipicolinic acid. AB - We use time-resolved coherent Raman spectroscopy to obtain molecule-specific signals from dipicolinic acid (DPA), which is a marker molecule for bacterial spores. We use femtosecond laser pulses in both visible and UV spectral regions and compare experimental results with theoretical predictions. By exciting vibrational coherence on more than one mode simultaneously, we observe a quantum beat signal that can be used to extract the parameters of molecular motion in DPA. The signal is enhanced when an UV probe pulse is used, because its frequency is near-resonant to the first excited electronic state of the molecule. The capability for unambiguous identification of DPA molecules will lead to a technique for real-time detection of spores. PMID- 16217022 TI - Regional vegetation die-off in response to global-change-type drought. AB - Future drought is projected to occur under warmer temperature conditions as climate change progresses, referred to here as global-change-type drought, yet quantitative assessments of the triggers and potential extent of drought-induced vegetation die-off remain pivotal uncertainties in assessing climate-change impacts. Of particular concern is regional-scale mortality of overstory trees, which rapidly alters ecosystem type, associated ecosystem properties, and land surface conditions for decades. Here, we quantify regional-scale vegetation die off across southwestern North American woodlands in 2002-2003 in response to drought and associated bark beetle infestations. At an intensively studied site within the region, we quantified that after 15 months of depleted soil water content, >90% of the dominant, overstory tree species (Pinus edulis, a pinon) died. The die-off was reflected in changes in a remotely sensed index of vegetation greenness (Normalized Difference Vegetation Index), not only at the intensively studied site but also across the region, extending over 12,000 km2 or more; aerial and field surveys confirmed the general extent of the die-off. Notably, the recent drought was warmer than the previous subcontinental drought of the 1950s. The limited, available observations suggest that die-off from the recent drought was more extensive than that from the previous drought, extending into wetter sites within the tree species' distribution. Our results quantify a trigger leading to rapid, drought-induced die-off of overstory woody plants at subcontinental scale and highlight the potential for such die-off to be more severe and extensive for future global-change-type drought under warmer conditions. PMID- 16217023 TI - Cenozoic continental climatic evolution of Central Europe. AB - Continental climate evolution of Central Europe has been reconstructed quantitatively for the last 45 million years providing inferred data on mean annual temperature and precipitation, and winter and summer temperatures. Although some regional effects occur, the European Cenozoic continental climate record correlates well with the global oxygen isotope record from marine environments. During the last 45 million years, continental cooling is especially pronounced for inferred winter temperatures but hardly observable from summer temperatures. Correspondingly, Cenozoic cooling in Central Europe is directly associated with an increase of seasonality. In contrast, inferred Cenozoic mean annual precipitation remained relatively stable, indicating the importance of latent heat transport throughout the Cenozoic. Moreover, our data support the concept that changes in atmospheric CO2 concentrations, although linked to climate changes, were not the major driving force of Cenozoic cooling. PMID- 16217024 TI - Dynamics of the acetylcholine receptor pore at the gating transition state. AB - Neuromuscular acetylcholine receptors (AChRs) are ion channels that alternatively adopt stable conformations that either allow (open) or prohibit (closed) ionic conduction. We probed the dynamics of pore (M2) residues at the diliganded gating transition state by using single-channel kinetic and rate-equilibrium free energy relationship (phi-value) analyses of mutant AChRs. The mutations were at the equatorial (9') position of the alpha, beta, and epsilon subunits (n = 15) or at sites between the equator and the extracellular domain in the alpha-subunit (n = 8). We also studied AChRs having only one of the two alpha-subunits mutated. The results indicate that the alpha-subunit, like the delta-subunit, has a region of flexure near the middle of M2, that the two alpha-subunits experience distinct energy barriers to gating at the equator (but not elsewhere), and that the collective subunit motions at the equator are asymmetric during the AChR gating isomerization. PMID- 16217025 TI - Severe muscle disease-causing desmin mutations interfere with in vitro filament assembly at distinct stages. AB - Desmin is the major intermediate filament (IF) protein of muscle. Recently, mutations of the desmin gene have been reported to cause familial or sporadic forms of human skeletal, as well as cardiac, myopathy, termed desmin-related myopathy (DRM). The impact of any of these mutations on filament assembly and integration into the cytoskeletal network of myocytes is currently not understood, despite the fact that all cause the same histopathological defect, i.e., desmin aggregation. To gain more insight into the molecular basis of this process, we investigated how mutations within the alpha-helical rod domain of desmin affect both the assembly of the recombinant protein in vitro as well as the filament-forming capacity in cDNA-transfected cells. Whereas 6 of 14 mutants assemble into seemingly normal IFs in the test tube, the other mutants interfere with the assembly process at distinct stages, i.e., tetramer formation, unit length filament (ULF) formation, filament elongation, and IF maturation. Correspondingly, the mutants with in vitro assembly defects yield dot-like aggregates in transfected cells, whereas the mutants that form IFs constitute a seemingly normal IF cytoskeleton in the cellular context. At present, it is entirely unclear why the latter mutant proteins also lead to aggregate formation in myocytes. Hence, these findings may be a starting point to dissect the contribution of the individual subdomains for desmin pathology and, eventually, the development of therapeutic interventions. PMID- 16217026 TI - Deleted in liver cancer 2 (DLC2) suppresses cell transformation by means of inhibition of RhoA activity. AB - The deleted in liver cancer 2 (DLC2) gene, located at chromosome 13q12.3, is a recently identified tumor suppressor gene. The gene is frequently underexpressed in human hepatocellular carcinoma, and its chromosomal region shows frequent deletion. DLC2 encodes a unique RhoGTPase-activating protein (RhoGAP) specific for small RhoGTPases, RhoA, and Cdc42. With bioinformatic analysis, we have identified four different isoforms of DLC2, which we named DLC2alpha, DLC2beta, DLC2gamma, and DLC2delta. Three of the isoforms contain the RhoGAP domain, namely, DLC2alpha, DLC2beta, and DLC2gamma. Ectopic expression of these three isoforms in mouse fibroblasts showed cytoplasmic localization. Of interest, overexpression of these isoforms suppressed the lysophosphatidic acid-induced stress fiber formation in mouse fibroblasts and changed the morphology of the transfected cells from angular and spindle to round. Furthermore, the RhoA pull down assay demonstrated a remarkable reduction in RhoA activity in the DLC2 transiently transfected cells. In contrast, cells transfected with inactive DLC2 GAP-mutant remained unchanged in cell morphology, actin stress fiber formation, and RhoA activity. HepG2 hepatoma cells stably transfected with the DLC2gamma isoform also changed to a round morphology, as in mouse fibroblasts. Of significance, these DLC2gamma stable transfectants showed marked suppression in cell proliferation, motility, and transformation, and there was a remarkable reduction in in vivo RhoA activity in these cells. These results suggest that DLC2 exhibits its tumor suppressor functions in vivo as a GAP specific for RhoA, exerting its effects in suppression of cytoskeleton reorganization, cell growth, cell migration, and transformation. PMID- 16217029 TI - Deep mantle structure and the postperovskite phase transition. AB - Seismologists have known for many years that the lowermost mantle of the Earth is complex. Models based on observed seismic phases sampling this region include relatively sharp horizontal discontinuities with strong zones of anisotropy, nearly vertical contrasts in structure, and small pockets of ultralow velocity zones (ULVZs). This diversity of structures is beginning to be understood in terms of geodynamics and mineral physics, with dense partial melts causing the ULVZs and a postperovskite solid-solid phase transition producing regional layering, with the possibility of large-scale variations in chemistry. This strong heterogeneity has significant implications on heat transport out of core, the evolution of the magnetic field, and magnetic field polarity reversals. PMID- 16217028 TI - Human infection patterns and heterogeneous exposure in river blindness. AB - Here we analyze patterns of human infection with Onchocerca volvulus (the cause of river blindness) in different continents and ecologies. In contrast with some geohelminths and schistosome parasites whose worm burdens typically exhibit a humped pattern with host age, patterns of O. volvulus infection vary markedly with locality. To test the hypothesis that such differences are partly due to heterogeneity in exposure to vector bites, we develop an age- and sex-structured model for intensity of infection, with parasite regulation within humans and vectors. The model is fitted to microfilarial data from savannah villages of northern Cameroon, coffee fincas of central Guatemala, and forest-dwelling communities of southern Venezuela that were recorded before introducing ivermectin treatment. Estimates of transmission and infection loads are compared with entomological and epidemiological field data. Host age- and sex heterogeneous exposure largely explains locale-specific infection patterns in onchocerciasis (whereas acquired protective immunity has been invoked for other helminth infections). The basic reproductive number, R0, ranges from 5 to 8, which is slightly above estimates for other helminth parasites but well below previously presented values. PMID- 16217027 TI - Crystal structures of oxidized and reduced mitochondrial thioredoxin reductase provide molecular details of the reaction mechanism. AB - Thioredoxin reductase (TrxR) is an essential enzyme required for the efficient maintenance of the cellular redox homeostasis, particularly in cancer cells that are sensitive to reactive oxygen species. In mammals, distinct isozymes function in the cytosol and mitochondria. Through an intricate mechanism, these enzymes transfer reducing equivalents from NADPH to bound FAD and subsequently to an active-site disulfide. In mammalian TrxRs, the dithiol then reduces a mobile C terminal selenocysteine-containing tetrapeptide of the opposing subunit of the dimer. Once activated, the C-terminal redox center reduces a disulfide bond within thioredoxin. In this report, we present the structural data on a mitochondrial TrxR, TrxR2 (also known as TR3 and TxnRd2). Mouse TrxR2, in which the essential selenocysteine residue had been replaced with cysteine, was isolated as a FAD-containing holoenzyme and crystallized (2.6 A; R = 22.2%; R(free) = 27.6%). The addition of NADPH to the TrxR2 crystals resulted in a color change, indicating reduction of the active-site disulfide and formation of a species presumed to be the flavin-thiolate charge transfer complex. Examination of the NADP(H)-bound model (3.0 A; R = 24.1%; R(free) = 31.2%) indicates that an active-site tyrosine residue must rotate from its initial position to stack against the nicotinamide ring of NADPH, which is juxtaposed to the isoalloxazine ring of FAD to facilitate hydride transfer. Detailed analysis of the structural data in conjunction with a model of the unusual C-terminal selenenylsulfide suggests molecular details of the reaction mechanism and highlights evolutionary adaptations among reductases. PMID- 16217030 TI - Gap junction-mediated intercellular biochemical coupling in cochlear supporting cells is required for normal cochlear functions. AB - Dysfunction of gap junctions (GJs) caused by mutations in connexin26 (Cx26) and Cx30 accounts for nearly half of all cases of hereditary nonsyndromic deafness cases. Although it is widely held that GJs connecting supporting cells in the organ of Corti mainly provide ionic pathways for rapid removal of K+ around the base of hair cells, the function of GJs in the cochlea remains unknown. Here we show that GJs were not assembled in the supporting cells of the organ of Corti until 3 days after birth in mice and then gradually matured to connect supporting cells before the onset of hearing. In organotypic cochlear cultures that were confirmed to express GJs, GJs mediated the propagation of intracellular Ca2+ concentration waves in supporting cells by allowing intercellular diffusion of inositol 1,4,5-trisphosphate. We found that a subset of structurally mild Cx26 mutations located at the second transmembrane region (V84L, V95M, and A88S) and a Cx30 mutation located at the first cytoplasmic segment (T5M) specifically affect the intercellular exchange of larger molecules but leave the ionic permeability intact. Our results indicated that Cx26 and Cx30 mutations that are linked to sensorineural deafness retained ionic coupling but were deficient in biochemical permeability. Therefore, GJ-mediated intercellular exchange of biochemically important molecules is required for normal cochlear functions. PMID- 16217031 TI - Low mobility of phosphatidylinositol 4,5-bisphosphate underlies receptor specificity of Gq-mediated ion channel regulation in atrial myocytes. AB - We have shown previously that cardiac G protein-gated inwardly rectifying K+ (GIRK) channels are inhibited by Gq protein-coupled receptors (GqPCRs) via phosphatidylinositol 4,5-bisphosphate (PIP2) depletion in a receptor-specific manner. To investigate the mechanism of receptor specificity, we examined whether the activation of GqPCRs induces localized PIP2 depletion. When we applied endothelin-1 to the bath, GIRK channel activities recorded in cell-attached patches were not changed, implying that PIP2 signal is not diffusible but is a localized signal. To test this possibility, we directly measured lateral diffusion by introducing fluorescence-labeled phosphoinositides to a small area of the membrane with patch pipettes. After pipettes were attached, phosphatidylinositol 4-monophosphate or phosphatidylinositol diffused rapidly to the entire membrane, whereas PIP2 was confined to the membrane patch inside the pipette. The confinement of PIP2 was disrupted after cytochalasin D treatment, suggesting that the cytoskeleton is responsible for the low mobility of PIP2. The diffusion coefficient (D) of PIP2 in the plasma membrane measured with the fluorescence recovery after photobleaching technique was 0.00039 microm2/s (n = 6), which is markedly lower than D of phosphatidylinositol (5.8 microm2/s, n = 5). Simulation of PIP2 concentration profiles by the diffusion model confirms that when D is small, the kinetics of PIP2 depletion at different distances from phospholipase C becomes similar to the characteristic kinetics of GIRK inhibition by different agonists. These results imply that PIP2 depletion is localized adjacent to GqPCRs because of its low mobility, and that spatial proximity of GqPCR and the target protein underlies the receptor specificity of PIP2-mediated signaling. PMID- 16217032 TI - BRIT1/MCPH1 is a DNA damage responsive protein that regulates the Brca1-Chk1 pathway, implicating checkpoint dysfunction in microcephaly. AB - BRIT1 [BRCT-repeat inhibitor of hTERT expression], a repressor of human telomerase function, is implicated in cellular immortalization. Here, we find that BRIT1 acts as a regulator of both the intra-S and G2/M checkpoints. When BRIT1 expression is depleted, cells lose the ionizing radiation (IR)-induced cell cycle arrest and become IR sensitive. BRIT1 is a chromatin-associated protein that forms irradiation-induced nuclear foci that colocalize with gamma-H2AX foci. BRIT1 is also required for the expression of both BRCA1 and the checkpoint kinase Chk1 and phosphorylation of Nbs1. Thus, the checkpoint defects in the absence of BRIT1 are likely to result from its regulation of Nbs1, BRCA1, and Chk1. BRIT1 is identical to the recently discovered MCPH1 gene, found mutant in patients with primary microcephaly. The ataxia telangiectasia mutated-Rad3 related (ATR)-Chk1 pathway is defective in Seckel syndrome, another microcephaly disorder. We propose that the microcephaly observed in patients with MCPH1 deficiencies is due to disruption of the ATR-BRCA1-Chk1 signaling pathway that is also disrupted in Seckel syndrome patients. PMID- 16217034 TI - Tectonic implications of Mars crustal magnetism. AB - Mars currently has no global magnetic field of internal origin but must have had one in the past, when the crust acquired intense magnetization, presumably by cooling in the presence of an Earth-like magnetic field (thermoremanent magnetization). A new map of the magnetic field of Mars, compiled by using measurements acquired at an approximately 400-km mapping altitude by the Mars Global Surveyor spacecraft, is presented here. The increased spatial resolution and sensitivity of this map provide new insight into the origin and evolution of the Mars crust. Variations in the crustal magnetic field appear in association with major faults, some previously identified in imagery and topography (Cerberus Rupes and Valles Marineris). Two parallel great faults are identified in Terra Meridiani by offset magnetic field contours. They appear similar to transform faults that occur in oceanic crust on Earth, and support the notion that the Mars crust formed during an early era of plate tectonics. PMID- 16217033 TI - RGS4 and RGS5 are in vivo substrates of the N-end rule pathway. AB - The ATE1-encoded Arg-transferase mediates conjugation of Arg to N-terminal Asp, Glu, and Cys of certain eukaryotic proteins, yielding N-terminal Arg that can act as a degradation signal for the ubiquitin-dependent N-end rule pathway. We have previously shown that mouse ATE1-/- embryos die with defects in heart development and angiogenesis. Here, we report that the ATE1 Arg-transferase mediates the in vivo degradation of RGS4 and RGS5, which are negative regulators of specific G proteins whose functions include cardiac growth and angiogenesis. The proteolysis of these regulators of G protein signaling (RGS) proteins was perturbed either by hypoxia or in cells lacking ubiquitin ligases UBR1 and/or UBR2. Mutant RGS proteins in which the conserved Cys-2 residue could not become N-terminal were long-lived in vivo. We propose a model in which the sequential modifications of RGS4, RGS5, and RGS16 (N-terminal exposure of their Cys-2, its oxidation, and subsequent arginylation) act as a licensing mechanism in response to extracellular and intracellular signals before the targeting for proteolysis by UBR1 and UBR2. We also show that ATE1-/- embryos are impaired in the activation of extracellular signal-regulated kinase mitogen-activated protein kinases and in the expression of G protein-induced downstream effectors such as Jun, cyclin D1, and beta-myosin heavy chain. These results establish RGS4 and RGS5 as in vivo substrates of the mammalian N-end rule pathway and also suggest that the O2-ATE1 UBR1/UBR2 proteolytic circuit plays a role in RGS-regulated G protein signaling in the cardiovascular system. PMID- 16217035 TI - A multifunctional enzyme adds to its repertoire. PMID- 16217036 TI - The opened processivity clamp slides into view. PMID- 16217037 TI - Profile of Akira Tonomura. PMID- 16217038 TI - Immunotherapy using unconjugated CD19 monoclonal antibodies in animal models for B lymphocyte malignancies and autoimmune disease. AB - Immunotherapy with unconjugated CD20 monoclonal antibodies has proven effective for treating non-Hodgkin's lymphoma and autoimmune disease. CD20 immunotherapy depletes mature B cells but does not effectively deplete pre-B or immature B cells, some B cell subpopulations, antibody-producing cells, or their malignant counterparts. Because CD19 is expressed earlier during B cell development, a therapeutic strategy for the treatment of early lymphoblastic leukemias/lymphomas was developed by using CD19-specific monoclonal antibodies in a transgenic mouse expressing human CD19. Pre-B cells and their malignant counterparts were depleted as well as antibody- and autoantibody-producing cells. These results demonstrate clinical utility for the treatment of diverse B cell malignancies, autoimmune disease, and humoral transplant rejection. PMID- 16217039 TI - Alteration of phosphatidylinositol 3-kinase cascade in the multilobulated nuclear formation of adult T cell leukemia/lymphoma (ATLL). AB - Adult T cell leukemia/lymphoma (ATLL) has been characterized as one of the most aggressive human neoplasias and its incidence is thought to be caused by both genetic and epigenetic alterations to the host cellular genes of T cells infected with human T cell leukemia virus type I (HTLV-I). A multilobulated nuclear appearance is an important diagnostic marker of ATLL, and we have now identified that the molecular mechanisms underlying these formations occur through microtubule rearrangement via phosphatidylinositol 3-kinase (PI3-kinase) activation by AILIM/ICOS signaling. We also show that PTEN and/or SHIP-1, which are PIP3 inositol phosphatases that inhibit the activation of downstream effectors of the PI3-kinase cascade, are disrupted in both ATLL neoplasias and in multilobulated nuclei-forming Jurkat cells. This down-regulation of PTEN was found to be essential for the formation of ATLL-type nuclear lobules. Furthermore, PI3-kinase and PTEN activities were observed to be closely associated with cellular proliferation. Thus, our results suggest that alteration of PI3-kinase signaling cascades, as a result of the down-regulation of inositol phosphatases, induces ATLL-type multilobulated nuclear formation and is also associated with the cellular proliferation of malignant T cell leukemias/lymphomas. PMID- 16217040 TI - Carbonic anhydrase isozyme-II-deficient mice lack the duodenal bicarbonate secretory response to prostaglandin E2. AB - Duodenal bicarbonate secretion (DBS) is accepted as the primary mucosal defense against acid discharged from the stomach and is impaired in patients with duodenal ulcer disease. The secretory response to luminal acid is the main physiological stimulus for DBS and involves mediation by PGE2 produced by mucosal cells. The aim of this investigation is to elucidate the role of carbonic anhydrases (CAs) II and IX in PGE2-mediated bicarbonate secretion in the murine duodenum. CA II- and IX-deficient mice and different combinations of their heterozygous and WT counterparts were studied. A 10-mm segment of the proximal duodenum with intact blood supply was isolated, and DBS was titrated by pH-stat (TitroLine-easy, Schott, Mainz, Germany). Mean arterial blood pressure (MAP) was continuously recorded, and blood acid/base balance and gastrointestinal morphology were analyzed. The duodenal segment spontaneously secreted HCO3(-) at a steady basal rate of 5.3 +/- 0.6 micromol x cm(-1) x h(-1). Perfusing the duodenal lumen for 20 min with 47 microM PGE2 caused a significant increase in DBS to 13.0 +/- 2.9 micromol x cm(-1) x h(-1), P < 0.0001. The DBS response to PGE2 was completely absent in Car2-/- mice, whereas basal DBS was normal. The CA IX-deficient mice with normal Car2 alleles showed a slight increase in DBS. Histological abnormalities were observed in the gastroduodenal epithelium in both CA II- and IX-deficient mice. Our data demonstrate a gastrointestinal phenotypic abnormality associated with CA II deficiency. The results show that the stimulatory effect of the duodenal secretagogue PGE2 completely depends on CA II. PMID- 16217042 TI - Generalized epileptic discharges show thalamocortical activation and suspension of the default state of the brain. AB - Our objective was to evaluate the brain regions showing increased and decreased metabolism in patients at the time of generalized bursts of epileptic discharges in order to understand their mechanism of generation and effect on brain function. By recording the electroencephalogram during the functional MRI, changes in the blood oxygenation level-dependent signal were obtained in response to epileptic discharges observed in the electroencephalogram of 15 patients with idiopathic generalized epilepsy. A group analysis was performed to determine the regions of positive (activation) and negative (deactivation) blood oxygenation level-dependent responses that were common to the patients. Activations were found bilaterally and symmetrically in the thalamus, mesial midfrontal region, insulae, and midline and bilateral cerebellum and on the borders of the lateral ventricles. Deactivations were bilateral and symmetrical in the anterior frontal and parietal regions and in the posterior cingulate gyri and were seen in the left posterior temporal region. Activations in thalamus and midfrontal regions confirm known involvement of these regions in the generation or spread of generalized epileptic discharges. Involvement of the insulae in generalized discharges had not previously been described. Cerebellar activation is not believed to reflect the generation of discharges. Deactivations in frontal and parietal regions remarkably followed the pattern of the default state of brain function. Thalamocortical activation and suspension of the default state may combine to cause the actual state of reduced responsiveness observed in patients during spike-and-wave discharges. This brief lapse of responsiveness may therefore not result only from the epileptic discharge but also from its effect on normal brain function. PMID- 16217041 TI - DNA and mRNA elements with complementary responses to hemin, antioxidant inducers, and iron control ferritin-L expression. AB - Ferritins, an ancient family of protein nanocages, concentrate iron in iron-oxy minerals for iron-protein biosynthesis and protection against oxy radical damage. Of the two genetic mechanisms that regulate rates of ferritin-L synthesis, DNA transcription and mRNA translation, more is known about mRNA regulation where iron targets complexes of an mRNA structure, the iron-responsive element (IRE) sequence, and ferritin IRE repressors (iron regulatory proteins 1 and 2). Neither the integration of mRNA and DNA regulation nor the ferritin-L DNA promoter are well studied. We now report the combined effects of DNA transcription and mRNA translation regulation of ferritin-L synthesis. First, the promoter of human ferritin-L, encoding the animal-specific subunit associated with human diseases, was identified, and contained an overlapping Maf recognition element (MARE) and antioxidant responsive element (ARE) that was positively regulated by tert butylhydroquinone, sulforaphane, and hemin with responses comparable to thioredoxin reductase (ARE regulator) or quinone reductase (MARE/ARE regulator). Iron, a poor regulator of the ferritin-L promoter, was 800 times less effective than sulforaphane. Combining the ferritin-L MARE/ARE and IRE produced a response to hemin that was 3-fold greater than the sum of responses of the MARE/ARE or IRE alone. Regulation of ferritin-L by a MARE/ARE DNA sequence emphasizes the importance of ferritin-L in oxidative stress that complements the mRNA regulation in iron stress. Combining DNA and mRNA mechanisms of regulation, as for ferritin L, illustrates the advantages of using two types of genetic targets to achieve sensitive responses to multiple signals. PMID- 16217043 TI - Unilateral toe-walking secondary to intramuscular hemangioma in the gastrocnemius. PMID- 16217045 TI - Does extracorporeal circulation harm the brain? PMID- 16217044 TI - Teaching NeuroImage: thromboembolic stroke in ICA stenosis. PMID- 16217046 TI - Neurologist-induced sexual dysfunction: enzyme-inducing antiepileptic drugs. PMID- 16217047 TI - Assessing the risk of a necessary harm: placebo surgery in Parkinson disease. PMID- 16217049 TI - Antiplatelet therapy contributes to acute deterioration of intracerebral hemorrhage. AB - OBJECTIVE: The purpose of this study was to examine the effect of antiplatelet therapy on the initial severity and the acute outcome of intracerebral hemorrhage (ICH). METHODS: The authors reviewed records of 251 consecutive patients hospitalized in their cerebrovascular center within 24 hours after onset of ICH. RESULTS: Fifty-seven patients (23%) had development of ICH during oral antiplatelet therapy. The major indication for antiplatelet therapy was the prevention of stroke recurrence (63%). As compared with patients without antiplatelet therapy, those who received antiplatelet therapy more frequently were aged 70 years or older (60% vs 35%; p < 0.001), had previous symptomatic ischemic stroke (54% vs 7%; p < 0.0001), had diabetes mellitus (26% vs 15%; p < 0.05), and had heart disease (32% vs 8%; p < 0.0001). Antiplatelet therapy was predictive of an increase in the hematoma volume by more than 40% on the second hospital day (hematoma enlargement, odds ratio [OR] 7.67, 95% CI 1.62 to 36.4) and the need for emergent surgical evacuation of the hematoma (OR 3.10, 95% CI 1.18 to 8.15). Antiplatelet therapy was an independent predictor for the occurrence of any of hematoma enlargement, emergent death, or evacuation surgery, which suggests that clinical deterioration occurs into the second hospital day (OR 7.45, 95% CI 2.46 to 22.5). CONCLUSIONS: Antiplatelet therapy seems to contribute to the acute clinical deterioration of intracerebral hemorrhage. PMID- 16217048 TI - Coronary artery bypass grafting is not a risk factor for dementia or Alzheimer disease. AB - OBJECTIVE: To study coronary artery bypass grafting (CABG) as a risk factor for dementia and Alzheimer disease (AD) using a case-control design. METHODS: The authors used the records-linkage system of the Rochester Epidemiology Project to ascertain incident cases of dementia in Rochester, MN, for the 5-year period 1990 to 1994. The authors defined dementia and AD using the criteria of the Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV). Each case was individually matched by age (+/-1 year) and sex to a person drawn randomly from the same population, and free of dementia in the index year (year of onset of dementia in the matched case). RESULTS: Among 557 dementia cases, 24 (4.3%) had undergone a CABG prior to the onset of dementia with a median lag time of 5.5 years (range = 0.1 to 15.9). Among 557 controls, 28 subjects (5.0%) had undergone a CABG prior to the index year with a median lag time 3.9 years (range = 0.1 to 12.3); OR = 0.85 (95% CI = 0.49 to 1.49; p = 0.57) for dementia and OR = 0.78 (95% CI = 0.39 to 1.56; p = 0.48) for AD. The findings did not change after adjustment for education. The perioperative courses of cases and controls were comparable. Analyses including only the 481 cases of dementia with presumed neurodegenerative or cerebrovascular etiology were also negative. CONCLUSIONS: This population-based case-control study suggests that coronary artery bypass grafting is not a major risk factor for dementia overall, or for Alzheimer disease. PMID- 16217050 TI - Retinal microvascular signs and risk of stroke and stroke mortality. AB - OBJECTIVE: The purpose of this study was to assess the relation of retinal microvascular signs and incident stroke and stroke mortality in an older population. METHODS: The authors took retinal photographs on baseline participants (3,654 patients aged 49+ years) of the Blue Mountains Eye Study (1992 to 1994). They assessed the presence of retinopathy (microaneurysms, retinal hemorrhages) in participants without diabetes and retinal arteriolar signs in all participants using standardized grading protocols. Incident stroke/TIA/cerebrovascular death (combined stroke events) were identified at follow-up examinations during 1997 to 1999. RESULTS: During a 7-year period, 859 participants died, 97 (11.3%) of which died of cerebrovascular causes. Of survivors, 24 had confirmed incident stroke, and 11 had incident TIA. Combined stroke events were more frequent in participants with retinopathy (5.7%), with moderate/severe arteriovenous nicking (4.2%), or with focal arteriolar narrowing (7.2%) compared with those without (1.9%). After controlling for age, sex, systolic blood pressure, smoking, and self-rated health, retinopathy was significantly associated with combined stroke events (relative risk [RR] 1.7, 95% CI 1.0 to 2.8) in persons without diabetes. This association was stronger in those without severe hypertension (RR 2.7, CI 1.2 to 6.2) or in persons with two or more retinal microvascular signs (RR 2.7, CI 1.5 to 5.2). Generalized or focal arteriolar narrowing or arteriovenous nicking was not independently associated with combined stroke events after multivariate adjustment. CONCLUSIONS: In older Australians without diabetes, retinopathy signs predict stroke or stroke-related death independent of traditional stroke risk factors. PMID- 16217051 TI - Aggressive dream content without daytime aggressiveness in REM sleep behavior disorder. AB - BACKGROUND: REM sleep behavior disorder (RBD) is characterized by vigorous sleep motor activity associated with dream mentation. Patients with RBD frequently report action-filled and violent dreams. OBJECTIVE: To systematically assess dream characteristics and daytime aggressiveness in patients with RBD and controls. METHODS: Forty-nine patients with polysomnographic-confirmed RBD diagnosis and 71 age- and sex-matched controls were asked to recall the most recent dreams and to complete the Aggression Questionnaire (AQ). Forty-one patients with RBD (81.6%; 36 men, 5 women; mean age: 67.5 +/- 7.5 years) and 35 controls (49.3%; 30 men, 5 women; mean age: 69.1 +/- 5.9 years) were able to remember their dreams and a total of 98 (RBD) and 69 (controls) dreams were collected in the two groups. Verbatim dream descriptions were scored and analyzed according to the Hall and Van De Castle method. RESULTS: Patients with RBD showed a higher percentage of dream with at least one aggressive episode (DWA) than controls (66% vs 15%; p < 0.00001), a higher aggression/friendliness interaction ratio (86% vs 44%; p < 0.0001), and a greater frequency of animal characters (19% vs 4%; p = 0.0001). In contrast to controls, no patient with RBD had dreams with elements of sexuality (0% vs 9%; p < 0.0001). The two groups did not differ in total AQ scores, except for a lower score on the physical aggressiveness subscale in patients with RBD compared to control subjects (16.5 +/- 6.4 vs 20.4 +/- 8.3; p = 0.034). No correlation was observed between dream aggressiveness and age, duration, or frequency of RBD symptoms. CONCLUSIONS: Dreams in patients with REM sleep behavior disorder were characterized by an elevated proportion of aggressive contents, despite normal levels of daytime aggressiveness. Dreams with aggressiveness and the known excessive phasic muscle activity during REM sleep may be related to the hyperactivity of a common neuronal generator. PMID- 16217052 TI - Differential effects of antiepileptic drugs on sexual function and hormones in men with epilepsy. AB - OBJECTIVE: To compare sexual function and reproductive hormone levels among men with epilepsy who took various antiepileptic drugs (AEDs), untreated men with epilepsy, and normal controls. METHODS: Subjects were 85 men with localization related epilepsy (25 on carbamazepine [CBZ], 25 on phenytoin [PHT], 25 on lamotrigine [LTG], and 10 untreated for at least 6 months [no AED]) and 25 controls. Sexual function scores (S-scores), hormone levels (bioactive testosterone, estradiol), hormone ratios (bioactive testosterone/bioactive estradiol), and gonadal efficiency (bioactive testosterone/luteinizing hormone) were compared among the five groups. RESULTS: S-scores, bioactive testosterone levels, bioactive testosterone/bioactive estradiol, and bioactive testosterone/luteinizing hormone were significantly greater in the control and LTG groups than in the CBZ and PHT groups. Sex hormone binding globulin was significantly higher in the CBZ and PHT groups than in all other groups. S-scores were below the control range in 20% of the men with epilepsy, including 32.0% on CBZ, 24% on PHT, 20% on no AEDs, and 4% on LTG (chi2: p = 0.08 for all four groups; chi2: p = 0.02 for the three AED groups). Bioactive testosterone was below the control range in 28.2%, including 48% on CBZ, 28% on PHT, 20% on no AEDs, and 12% on LTG (chi2: p = 0.02). Among men with epilepsy who had low S scores, 70.6% had bioactive testosterone levels below the control range as compared to 17.6% among men with normal S-scores (chi2: p < 0.0001). Among men with epilepsy who had abnormally low bioactive testosterone, 50.0% had low S scores; among men with normal bioactive testosterone, 8.2% had low S-scores (chi2: p < 0.0001). Bioactive testosterone decline with age was significantly greater among men with epilepsy than among controls and notably greater in the CBZ and PHT groups than in the LTG and untreated groups. CONCLUSIONS: Sexual function, bioavailable testosterone levels, and gonadal efficiency in men with epilepsy who took lamotrigine were comparable to control and untreated values and significantly greater than with carbamazepine or phenytoin treatment. PMID- 16217053 TI - Preoperative heart rate variability in relation to surgery outcome in refractory epilepsy. AB - BACKGROUND: Epilepsy patients may have an impaired autonomic cardiac control, which has been associated with an increased incidence of sudden unexpected death among people with epilepsy (SUDEP). The risk of SUDEP is particularly high among epilepsy surgery candidates with refractory epilepsy. This risk seems to be reduced after successful surgery but whether this is an effect of surgery or reflects pre-existing differences between good and poor responders is under debate. METHODS: We used spectral analysis to analyze prospectively heart rate variability (HRV) preoperatively in 21 consecutive patients with temporal lobe epilepsy who were planned for epilepsy surgery. The presurgical HRV based on ambulatory 24 hours EKG recordings was analyzed in relation to seizure control at 1 year after surgery. RESULTS: Patients had significantly lower SD of RR intervals, total power, very low frequency power and low frequency power than matched healthy controls. Patients with good outcome of surgery (Engel class I; n = 11) did not differ from their controls while those with poor outcome (Engel class II-IV; n = 10) had significantly lower power in all domains than those with a favorable outcome. CONCLUSIONS: Measurements of heart rate variability preoperatively indicate that patients with a poor outcome of surgery have a more pronounced impairment of sympathetic as well as parasympathetic cardiac control than those with good outcome. Reduced heart rate variability may be associated with an increased risk of sudden unexpected death among people with epilepsy (SUDEP). Good surgery candidates may a priori have a lower risk of SUDEP. PMID- 16217054 TI - 3T phased array MRI improves the presurgical evaluation in focal epilepsies: a prospective study. AB - BACKGROUND: Although detection of concordant lesions on MRI significantly improves postsurgical outcomes in focal epilepsy (FE), many conventional MR studies remain negative. The authors evaluated the role of phased array surface coil studies performed at 3 Tesla (3T PA MRI). METHODS: Forty patients with medically intractable focal epilepsies were prospectively imaged with 3T PA-MRI including high matrix TSE T2, fluid attenuated inversion recovery, and magnetization prepared rapid gradient echo. All patients were considered candidates for epilepsy surgery. 3T PA-MRIs were reviewed by a neuroradiologist experienced in epilepsy imaging with access to clinical information. Findings were compared to reports of prior standard 1.5T MRI epilepsy studies performed at tertiary care centers. RESULTS: Experienced, unblinded review of 3T PA-MRI studies yielded additional diagnostic information in 48% (19/40) compared to routine clinical reads at 1.5T. In 37.5% (15/40), this additional information motivated a change in clinical management. In the subgroup of patients with prior 1.5T MRIs interpreted as normal, 3T PA-MRI resulted in the detection of a new lesion in 65% (15/23). In the subgroup of 15 patients with known lesions, 3T PA MRI better defined the lesion in 33% (5/15). CONCLUSION: Phased array surface coil studies performed at 3 Tesla read by an experienced unblinded neuroradiologist can improve the presurgical evaluation of patients with focal epilepsy when compared to routine clinical 1.5T studies read at tertiary care centers. PMID- 16217055 TI - Transient lesion in the splenium of the corpus callosum and antiepileptic drug withdrawal. AB - OBJECTIVE: To test pathophysiologic hypotheses regarding the occurrence of a splenial lesion in patients with epilepsy. METHODS: The authors studied 16 patients with a splenial lesion and 32 control patients, all of whom had MRI examination immediately after presurgical EEG long-term monitoring (LTM). The authors compared the number of generalized tonic-clonic and partial seizures during LTM, antiepileptic drug (AED) withdrawal, and laboratory results. RESULTS: All of the patients with a splenial lesion had their AEDs stopped completely, vs 47% of the controls (p = 0.001). Patients with SCC lesion had a longer duration of complete withdrawal (median 3.5 vs 2 days, p = 0.03). There was no correlation with seizure frequency or the introduction of new AEDs. CONCLUSION: A lesion of the splenium of the corpus callosum in patients with epilepsy is not associated with toxic drug effects or high seizure frequency, but might be induced by a rapid and relatively long-lasting reduction of antiepileptic drugs. Its frequency might be underestimated as MRI after long-term monitoring is rarely done. PMID- 16217056 TI - The nature and frequency of cognitive deficits in children with neurofibromatosis type 1. AB - OBJECTIVE: To assess the frequency and severity of specific cognitive deficits in children with neurofibromatosis type 1 (NF1) in a large unbiased cohort. METHODS: Extensive cognitive assessments were performed in 81 children with NF1 ages 8 to 16 years and their performance was compared with that of 49 unaffected sibling controls. RESULTS: Eighty-one percent of the children with NF1 had moderate to severe impairment in one or more areas of cognitive functioning. Although 51% of children with NF1 performed poorly on tasks of reading, spelling, and mathematics, specific learning disabilities (as defined by IQ-achievement discrepancies) were present in only 20% of the children. Sustained attention difficulties were present in 63% of children with NF1, with 38% of children with NF1 fulfilling the diagnostic criteria for attention deficit-hyperactivity disorder. The NF1 neuropsychological profile is characterized by deficits in perceptual skills (visuospatial and visuoperceptual), executive functioning (planning and abstract concept formation), and attention (sustained and switching). Interestingly, both verbal and visual memory was unaffected in NF1 children, and their memory skills were in general stronger than their level of general intellectual function. Although both expressive and receptive language skills were significantly impaired in NF1 children, they appeared to be relatively better preserved than visuospatial abilities once IQ is taken into account. CONCLUSION: There is an extremely high frequency of cognitive problems in children with neurofibromatosis type 1, making cognitive dysfunction the most common complication to affect quality of life in these children. PMID- 16217057 TI - Serum cholesterol and risk of Alzheimer disease: a community-based cohort study. AB - OBJECTIVES: To examine the association of serum total cholesterol (TC) and high density lipoprotein (HDL) levels and subsequent incidence of dementia and Alzheimer disease (AD) in a population-based cohort study. METHODS: A cohort of cognitively intact persons, aged 65 and older, was randomly selected from Group Health Cooperative (GHC), a large health maintenance organization, and was assessed biennially for dementia. Premorbid levels of TC and HDL were obtained from a computerized clinical laboratory database at GHC. Cox proportional hazards regression was used to calculate hazard ratios (HR, 95% CI) for dementia and AD associated with quartiles of TC and HDL levels. RESULTS: Of the 2,356 eligible participants, 2,141 had at least one serum TC measure prior to the initial enrollment. Using the lowest TC quartiles as the reference group, the HR in the highest TC quartiles was not significantly elevated for dementia (1.16, 0.81 to 1.67) or for AD (1.00, 0.61 to 1.62) after adjusting for age, sex, education, baseline cognition, vascular comorbidities, body mass index, and lipid-lowering agent use. Serum HDL showed a similar lack of significant association with risk of dementia or AD. Models that included the presence of one or more APOE-epsilon4 alleles showed a typical association of epsilon4 with AD risk. This association was not materially modified by inclusion of TC level. CONCLUSION: The data do not support an association between serum total cholesterol or high density lipoprotein in late life and subsequent risk of dementia or Alzheimer disease (AD). The increased risk of AD with APOE-epsilon4 is probably not mediated by serum total cholesterol levels. PMID- 16217058 TI - Neuroradiologic and clinicopathologic features of oculoleptomeningeal type amyloidosis. AB - OBJECTIVE: To clarify the pathogenesis of leptomeningeal amyloidosis in familial amyloidotic polyneuropathy amyloidogenic transthyretin Y114C (FAP ATTR Y114C). METHODS: The authors analyzed eight FAP ATTR Y114C patients. Six patients showed CNS symptoms associated with leptomeningeal amyloidosis. To examine the function of the blood-CSF barrier and blood-brain barrier (BBB), the authors performed CSF and MRI studies. The authors also performed a histopathologic study of autopsy specimens to examine the distribution of amyloid deposition in the CNS. RESULTS: CSF study showed high total protein concentrations and increased albumin CSF/serum concentration quotients (Qalb; an indication of blood-CSF barrier function). MRI with gadolinium (Gd) revealed enhancement from brainstem to spinal cord. Serial brain MRI studies with FLAIR images after Gd administration showed Gd leakage into the subarachnoid space (two patients). These findings suggested the blood-CSF barrier and BBB dysfunctions. Constructive interference in steady state (CISS) three-dimensional Fourier transformation (CISS-3DFT) sequence analysis demonstrated amyloid-induced funiculus structures joining the spinal cord and dura mater (one patient). Histopathologic study revealed intense amyloid deposition in leptomeninges, vessel walls, and parenchyma in spinal cord and the brain. These distributions of amyloid deposition are unique compared to other TTR related leptomeningeal amyloidosis. CONCLUSIONS: Patients with familial amyloidotic polyneuropathy amyloidogenic transthyretin Y114C had CNS disorders related to amyloid deposition in leptomeninges, vessel walls, and parenchyma in spinal cord and the brain. The pathogenesis of CNS disorders may reflect disruption of the blood-CSF barrier and blood-brain barrier by amyloid deposition. PMID- 16217059 TI - Postinfectious inflammatory disorders: subgroups based on prospective follow-up. AB - BACKGROUND: Acute disseminated encephalomyelitis (ADEM) refers to a monophasic acute multifocal inflammatory CNS disease. However, both relapsing and site restricted variants, possibly associated with peripheral nervous system (PNS) involvement, are also observed, and a systematic classification is lacking. OBJECTIVE: To describe a cohort of postinfectious ADEM patients, to propose a classification based on clinical and instrumental features, and to identify subgroups of patients with different prognostic factors. METHODS: Inpatients of a Neurologic and Infectious Disease Clinic affected by postinfectious CNS syndrome consecutively admitted over 5 years were studied. RESULTS: Of 75 patients enrolled, 60 fulfilled criteria for ADEM after follow-up lasting from 24 months to 7 years. Based on lesion distribution, patients were classified as encephalitis (20%), myelitis (23.3%), encephalomyelitis (13.3%), encephalomyeloradiculoneuritis (26.7%), and myeloradiculoneuritis (16.7%). Thirty patients (50%) had a favorable outcome. Fifteen patients (25%) showed a relapsing course. Poor outcome was related with older age at onset, female gender, elevated CSF proteins, and spinal cord and PNS involvement. All but two patients received high-dose steroids as first-line treatment, with a positive response in 39 (67%). Ten of 19 nonresponders (53%) benefited from high-dose IV immunoglobulin; 9 of 10 had PNS involvement. The data were not controlled. CONCLUSIONS: A high prevalence of "atypical variants" was found in this series, with site-restricted damage or additional peripheral nervous system (PNS) involvement. Prognosis and response to steroids were generally good, except for some patient subgroups. In patients with PNS involvement and steroid failure, a favorable effect of IV immunoglobulin was observed. PMID- 16217060 TI - Changes in the ascertainment of multiple sclerosis. AB - OBJECTIVE: With diagnostic criteria alterations, increased MRI availability, and awareness of therapies, temporal changes in incidence and prevalence rates may occur, with an increase in the proportion of mildly affected persons diagnosed with multiple sclerosis (MS). The authors assessed temporal trends in the delay from symptom onset to diagnosis (DONDX), and determined whether the degree of disability at diagnosis differs by year of symptom onset (YONSET), using the NARCOMS Registry. METHODS: The authors selected US participants with an age at symptom onset of 10 to 60 years, and YONSET > or = 1980 (n = 16,581). The authors divided YONSET into 5-year groups and compared DONDX between groups using multivariate Cox regression. The authors classified participants enrolled within 2 years of diagnosis (n = 5,548) as having mild, moderate, or severe disability using Patient Determined Disease Steps, and assessed the association of disability with YONSET using polytomous logistic regression. RESULTS: DONDX decreased with later YONSET (r = -0.43, p < 0.0001). This association remained after adjustment for demographic factors in a multivariate Cox model. Later YONSET was associated with increased odds of having mild disability at diagnosis as compared to severe disability (OR = 1.10 per year; 1.09 to 1.11). CONCLUSION: The delay from symptom onset to diagnosis is steadily decreasing in MS. An increasing proportion of patients with MS have mild disability at diagnosis after accounting for confounders. As the effectiveness of therapies is influenced by disease duration, this has implications for comparison of treatment effects in modern clinical trials to earlier study results. PMID- 16217061 TI - MRI as a marker for disease heterogeneity in multiple sclerosis. AB - BACKGROUND: Whereas recent data from imaging studies challenge the prevailing notion that multiple sclerosis (MS) is purely an inflammatory disease, pathologic studies suggest differences in the disease processes between individual patients with MS. The ability to dissect the pathophysiologic disease heterogeneity, if it indeed exists, by methodologies that can be applied in vivo is important both for the development of new therapeutics and for the ability to identify the optimal therapy for an individual patient. OBJECTIVE: To design a stratification algorithm for patients with MS based on accepted MRI measurements reflective of inflammation and axonal damage/tissue loss and to assess if such MS subgroups retain their intergroup differences long term. METHODS: Mathematical modeling was used to select three discriminatory MRI measures for clinical outcome based on the cross-sectional analysis of 71 patients with untreated MS and tested general applicability of the stratification scheme on the independent longitudinal cohort of 71 MS patients. RESULTS: By consecutive employment of MRI measures reflective of inflammation and tissue loss, the authors were able to separate MS patients into four clinically meaningful subgroups. The analysis of the longitudinal confirmatory cohort demonstrated persistence of the intergroup differences in selected MRI measures for 8 years. CONCLUSIONS: The inflammatory activity and destructiveness of the multiple sclerosis process are to some degree independent of each other, and the successive evaluation of both of these variables can strengthen prediction of clinical outcome in individual patients. PMID- 16217062 TI - IL-1RN and IL-1B gene polymorphisms and cerebral hemorrhagic events after traumatic brain injury. AB - OBJECTIVE: To investigate the association of (variable number tandem repeat) interleukin (IL) 1RN and (-511) IL-1B gene polymorphisms with brain hemorrhagic events after traumatic brain injury (TBI). METHODS: Data from brain CT, Glasgow Coma Scale (GCS) at admission, and 6-month Glasgow Outcome Scale (GOS) and modified Rankin Scale (mRS) were collected for 151 prospectively recruited patients with TBI. IL-1RN and IL-1B genotypes were determined using standard methods. Presence vs absence of any type of brain hemorrhage was the main outcome. Type of brain hemorrhage, GCS at admission, and 6-month GOS and mRS were secondary outcomes. Odd ratios (ORs) and corresponding 95% CI were calculated using logistic regression analyses. In adjusted models, the associations were controlled for age, gender, diffuse brain edema, volume of intracranial hematoma, neurosurgical intervention, and GCS at admission. p values less than 0.01 were considered significant. RESULTS: Compared with noncarriers, IL-1RN allele 2 carriers had higher odds of having cerebral hemorrhages after TBI (adjusted OR = 4.57; 95% CI = 1.67 to 12.96; p = 0.004). The associations for (-511) IL-1B polymorphism were not significant. CONCLUSION: There is an association between the presence of interleukin-1RN allele 2 and posttraumatic brain hemorrhage. PMID- 16217064 TI - Transverse sinus stenoses persist after normalization of the CSF pressure in IIH. AB - BACKGROUND: Bilateral transverse sinus (TS) stenosis has been found in more than 90% of patients with idiopathic intracranial hypertension (IIH). OBJECTIVE: To evaluate whether TS stenosis changed after normalization of CSF pressure in patients with IIH during medical treatment. METHODS: Fourteen consecutive patients with IIH with bilateral TS stenosis on cerebral MR venography (MRV) during the medical treatment were studied. Patients were followed for over a 6 year period. During the follow-up, patients underwent repeated lumbar punctures (LPs) and cerebral MRV. MRV was always performed before each LP. RESULTS: TS stenosis persisted in all the patients during the follow-up. In 9 of 14 (64%) patients with IIH, CSF pressure normalized during medical treatment. CONCLUSIONS: Transverse sinus (TS) stenoses, as revealed by MR venography, persist in patients with idiopathic intracranial hypertension after normalization of CSF pressure, suggesting the lack of a direct relationship between the caliber of TS and CSF pressure. PMID- 16217063 TI - Andersen-Tawil syndrome: new potassium channel mutations and possible phenotypic variation. AB - OBJECTIVE: To evaluate clinical, genetic, and electrophysiologic features of patients with Andersen-Tawil syndrome (ATS) in the United Kingdom. METHODS: Clinical and neurophysiologic evaluation was conducted of 11 families suspected to have ATS. Molecular genetic analysis of each proband was performed by direct DNA sequencing of the entire coding region of KCNJ2. Control samples were screened by direct DNA sequencing. The electrophysiologic consequences of several new mutations were studied in an oocyte expression system. RESULTS: All 11 ATS families harbored pathogenic mutations in KCNJ2 with six mutations not previously reported. Some unusual clinical features including renal tubular defect, CNS involvement, and dental and phonation abnormalities were observed. Five mutations (T75M, D78G, R82Q, L217P, and G300D) were expressed, all of which resulted in nonfunctional channels when expressed alone, and co-expression with wild-type (WT) KCNJ2 demonstrated a dominant negative effect. CONCLUSION: Six new disease causing mutations in KCNJ2 were identified, one of which was in a PIP2 binding site. Molecular expression studies indicated that five of the mutations exerted a dominant negative effect on the wild-type allele. KCNJ2 mutations are an important cause of ATS in the UK. PMID- 16217065 TI - Three-dimensional preoperative maps of hippocampal atrophy predict surgical outcomes in temporal lobe epilepsy. AB - The authors used surface-based anatomic mapping to detect features of hippocampal anatomy that correlated with surgical outcomes in patients undergoing surgery for mesial temporal lobe epilepsy with hippocampal sclerosis. Compared with a seizure free group, hippocampal profiles for the non-seizure-free group had greater diffuse ipsilateral atrophy and more region-specific contralateral atrophy in the anterior, lateral hippocampus. These atrophic regions may indicate areas of increased epileptogenicity, contributing to poorer surgical outcomes. PMID- 16217066 TI - Clinical features of benign infantile convulsions: familial and sporadic cases. AB - The authors describe the so-called benign convulsions of infancy and confirm the existence of benign nonfamilial infantile convulsions during the first 2 years of life and their benign course. The authors evaluated 58 patients: 17 subjects had a family history of benign epilepsy, and 41 did not. No clinical differences were observed between the two groups. PMID- 16217067 TI - L-DOPS therapy for refractory orthostatic hypotension in autoimmune autonomic neuropathy. AB - The authors report a 46-year-old woman with antibodies to the nicotinic acetylcholine receptor (NiAchR) of the autonomic ganglia. She presented with severe orthostatic intolerance refractory to treatment with midodrine, fludrocortisone, erythropoietin, vasopressin, salt, and fluid loading. Addition of L-threo-3,4-dihidroxyphenylserine (L-DOPS) substantially improved blood pressure and orthostatic tolerance. L-DOPS may benefit patients with severe orthostatic intolerance and be particularly effective in patients with ganglionic NiAchR antibodies. PMID- 16217068 TI - ERG and anatomical abnormalities suggesting retinopathy in dementia with Lewy bodies. AB - The authors used flash electroretinography to demonstrate dysfunction of the photopic and scotopic retina in patients with dementia with Lewy bodies and visual hallucinations (VHs) compared with patients with Parkinson disease, patients without VHs, and controls. The retinal dysfunction may be related to slight alteration of the photoreceptors and numerous pale inclusions in the outer plexiform layer found at the post mortem examination, suggesting a specific retinopathy. PMID- 16217069 TI - Kjellin syndrome: first case with retinal changes in carriers. PMID- 16217070 TI - Autoantibodies in postinfectious acute cerebellar ataxia. AB - The authors found serum immunoglobulin M (IgM) autoantibody in a patient with typical acute cerebellar ataxia (ACA) and identified the antigen molecule as triosephosphate isomerase (TPI). TPI antigenicity to the patient's antibody was the highest in the cerebellar tissue. Eight of 23 patients with ACA had increased IgM anti-TPI antibody titers vs those of healthy controls. Preceding Epstein-Barr virus infection was confirmed serologically in all 8 patients. Anti-TPI antibody decreased with clinical improvement. PMID- 16217071 TI - Reality confusion in spontaneous confabulation. AB - A woman produced spontaneous confabulations after rupture of an anterior communicating artery aneurysm. She confused currently irrelevant with currently relevant information in implicit memory; confabulations about people concerned only new acquaintances; false reality could be induced by an intensive 5-minute discussion; and in a recognition task, she confused false repetitions in another modality with real item repetitions. The findings support the theory that the defect causing spontaneous confabulation precedes conscious memory processing. PMID- 16217072 TI - CSF levels of angiogenesis-related proteins in patients with leptomeningeal metastases. AB - The authors determined the levels of vascular endothelial growth factor (VEGF) and urokinase-type plasminogen activator (uPA) in the CSF of patients with leptomeningeal metastases (LM; n = 53), cancer patients without LM (n = 18), and subjects without malignancy (n = 25). Median levels of uPA and VEGF were significantly higher in patients with LM, supporting the hypothesis that angiogenesis contributes to LM. VEGF was negatively correlated with survival in patients with LM, suggesting its use as a prognostic factor. PMID- 16217073 TI - Psychotic disorders in MS: population-based evidence of an association. AB - Administrative data from a Canadian province were accessed to assess the association of psychotic disorders with multiple sclerosis (MS). Physician assigned ICD-9-CM codes were used to classify MS and psychotic disorder status. A significant association was observed. Additional research is needed to determine the clinical importance and treatment needs of this population. PMID- 16217074 TI - Evidence for progressive gray matter loss in patients with relapsing-remitting MS. AB - Little is known about the temporal evolution of gray matter damage occurring early in the course of multiple sclerosis (MS). The authors investigated the evolution of gray matter volume loss in 117 patients with relapsing-remitting MS, scanned monthly for a 9-month period. Time-trend analysis revealed a decrease of gray matter volumes over the study period (p < 0.001). This study shows that gray matter damage in relapsing-remitting MS evolves markedly over a short period of observation. PMID- 16217075 TI - Primary central nervous system lymphomas (PCNSL): MRI response criteria revised. AB - The authors investigated the applicability of Macdonald response criteria to patients with primary CNS lymphoma (PCNSL). Four of 68 patients with persisting contrast-enhancing lesions after primary therapy did not receive further therapy, and none showed tumor progression after up to 54 months. Because contrast enhancement may not necessarily indicate residual, biologically active lymphoma, the authors propose a modification of the Macdonald response criteria. PMID- 16217076 TI - A new congenital form of X-linked autophagic vacuolar myopathy. AB - In a new family with X-linked congenital autophagic vacuolar myopathy (AVM), seven affected boys presented with congenital hypotonia, dyspnea, and dysphagia with delayed motor milestones. Muscle pathology revealed autophagic vacuoles with sarcolemmal features, multilayered basal lamina with marked sarcolemmal deposition of C5-9 membrane attack complex and calcium, histologically indistinguishable from childhood-onset X-linked myopathy with excessive autophagy (XMEA). Haplotype analysis suggests that this new AVM and XMEA may be allelic despite different clinical presentations. PMID- 16217077 TI - Isolated left vein of Labbe thrombosis. PMID- 16217078 TI - Pulmonary embolism: an unusual complication of cerebral venous thrombosis. PMID- 16217079 TI - Herbal energy drinks, phenylpropanoid compounds, and cerebral vasculopathy. PMID- 16217080 TI - Cervical epidural blood patch for low CSF pressure headaches. PMID- 16217081 TI - Age, dose, and environmental temperature are risk factors for topiramate-related hyperthermia. PMID- 16217083 TI - Fiction: Smug little men. PMID- 16217082 TI - Compressive myelopathy presenting as cervical cord neurapraxia: a differential diagnosis of TIA. PMID- 16217084 TI - The role of radiotracer imaging in Parkinson disease. PMID- 16217085 TI - Subacute sclerosing panencephalitis in the differential diagnosis of encephalitis. PMID- 16217086 TI - Cerebrovascular perfusion in marijuana users during a month of monitored abstinence. PMID- 16217087 TI - Strokes in the subinsular territory: clinical, topographical, and etiological patterns. PMID- 16217088 TI - Katrina, climate change and the poor. PMID- 16217089 TI - The marginalization of some medical students. PMID- 16217091 TI - Excluding the experts? PMID- 16217093 TI - An unusual crystal. PMID- 16217095 TI - Coxibs and cardiovascular risk. PMID- 16217098 TI - Coxibs and cardiovascular risk. PMID- 16217096 TI - Coxibs and cardiovascular risk. PMID- 16217099 TI - Physicians want transparency as Guidant lawsuits grow. PMID- 16217101 TI - Med schools need to train more doctors: Dosanjh. PMID- 16217102 TI - Inconclusive evidence puts Adderall back on the market. PMID- 16217103 TI - Oxidative stress in ocular disease: does lutein play a protective role? PMID- 16217104 TI - Link between porphyria and fasting uncovered. PMID- 16217105 TI - Pathogens fight back. PMID- 16217106 TI - Applying the 2005 Canadian Hypertension Education Program recommendations: 4. Managing uncomplicated hypertension. PMID- 16217107 TI - The changing ecology of avian flu. PMID- 16217108 TI - A newborn requiring selective bronchial intubation. PMID- 16217109 TI - Does beta-blocker prophylaxis improve survival after major noncardiac surgery? PMID- 16217110 TI - Cost-effectiveness of computerized tomographic colonography versus colonoscopy for colorectal cancer screening. AB - BACKGROUND: Computerized tomographic (CT) colonography is a potential alternative to colonoscopy for colorectal cancer screening. Its main advantage, a better safety profile, may be offset by its limitations: lower sensitivity, need for colonoscopy in cases where results are positive, and expense. METHODS: We performed an economic evaluation, using decision analysis, to compare CT colonography with colonoscopy for colorectal cancer screening in patients over 50 years of age. Three-year outcomes included number of colonoscopies, perforations and adenomas removed; deaths from perforation and from colorectal cancer from missed adenomas; and direct health care costs. The expected prevalence of adenomas, test performance characteristics of CT colonography and colonoscopy, and probability of colonoscopy complications and cancer from missed adenomas were derived from the literature. Costs were determined in detail locally. RESULTS: Using the base-case assumptions, a strategy of CT colonography for colorectal cancer screening would cost 2.27 million dollars extra per 100,000 patients screened; 3.78 perforation-related deaths would be avoided, but 4.11 extra deaths would occur from missed adenomas. Because screening with CT colonography would cost more and result in more deaths overall compared with colonoscopy, the latter remained the dominant strategy. Our results were sensitive to CT colonography's test performance characteristics, the malignant risk of missed adenomas, the risk of perforation and related death, the procedural costs and differences in screening adherence. INTERPRETATION: At present, CT colonography cannot be recommended as a primary means of population-based colorectal cancer screening in Canada. PMID- 16217111 TI - Cigarette smoking and cardiovascular risk factors among Aboriginal Canadian youths. AB - BACKGROUND: Aboriginal populations in North America are exhibiting an increased prevalence of cardiovascular disease and associated traditional and nontraditional cardiovascular risk factors, trends believed to be due to the influence of Western lifestyle habits. Because these influences are present at an early age, we sought to study the patterns of one such habit, cigarette smoking, among Aboriginal Canadian youths and to assess the associated accrual of cardiovascular risk factors at an early age. METHODS: Patterns of cigarette smoking were assessed in a population-based, cross-sectional study involving 236 youths aged 10-19 (mean 14.9) years in the Oji-Cree community of Sandy lake, in northwestern Ontario. Participants underwent clinical and metabolic evaluation with assessment of cardiovascular risk factors. RESULTS: The prevalence of cigarette smoking among the study participants was considerably higher than age specific national averages, with fully 50% of the participants overall and 82% of the adolescent participants (aged 15-19) being current smokers. Compared with their peers, children smoking 6 or more cigarettes per day had an enhanced cardiovascular risk profile consisting of a higher mean systolic blood pressure (111 v. 107.5 mm Hg, p = 0.036), a higher mean plasma homocysteine level (8.7 v. 7.6 micromol/L, p = 0.008) and a lower mean serum folate level (4.5 v. 5.4 mmol/L, p = 0.007), after adjustment for age, sex and body mass index. In separate multiple linear regression analyses, current cigarette exposure (number of cigarettes smoked per day) emerged as an independent determinant of both systolic blood pressure and plasma homocysteine level. INTERPRETATION: In this Aboriginal community with remarkably high rates of cigarette smoking among its youth, an independent dose-response relation was found between current smoking exposure and both traditional (systolic blood pressure) and nontraditional (homocysteine level) cardiovascular risk factors. The association of cigarette smoking with an enhanced cardiovascular risk profile at an early age may be a factor contributing to the high prevalence of cardiovascular disease in this Aboriginal population. PMID- 16217112 TI - Improving patient safety: moving beyond the "hype" of medical errors. PMID- 16217113 TI - The Chaoulli decision: a debate in which physicians must be heard. PMID- 16217114 TI - Public versus private: the medical resident perspective. PMID- 16217115 TI - Should Canadian physicians support parallel private health care? PMID- 16217117 TI - Mutable influences on intimate partner abuse screening in health care settings: a synthesis of the literature. AB - For more than a decade, professional medical and nursing associations have recommended universal screening for intimate partner abuse (IPA). This review of 44 studies examines IPA screening during this period. The purposes of the review are to (a) identify mutable influences on IPA screening, (b) summarize what is known about altering these influences, and (c) outline an agenda for improving IPA screening. Evidence to date indicates that screening is not universal. Promising approaches to improvement start with provider training and relatively simple environmental adjustments. Unanswered empirical questions include "What are the essential components of effective training? Which health care providers are best positioned to screen under which circumstances? Which of the untested mutable influences will further improve IPA screening practice?" The review leads to the following practice recommendations: (a) Interventions should focus on clinicians in training (e.g., medical residents), and (b) screening approaches must be tailored to various practice settings. PMID- 16217116 TI - Diagnosis and treatment of ectopic pregnancy. AB - Ectopic pregnancy is a life- and fertility-threatening condition that is commonly seen in Canadian emergency departments. Increases in the availability and use of hormonal markers, coupled with advances in formal and emergency ultrasonography have changed the diagnostic approach to the patient in the emergency department with first-trimester bleeding or pain. Ultrasonography should be the initial investigation for symptomatic women in their first trimester; when the results are indeterminate, the serum beta human chorionic gonadotropin (beta-hCG) concentration should be measured. Serial measurement of beta-hCG and progesterone concentrations may be useful when the diagnosis remains unclear. Advances in surgical and medical therapy for ectopic pregnancy have allowed the proliferation of minimally invasive or noninvasive treatment. Guidelines for laparoscopy and for methotrexate therapy are provided. PMID- 16217118 TI - Intimate partner violence, employment,and the workplace: consequences and future directions. AB - The purpose of this article is to examine the literature on violence against women and employment. After a brief discussion of the definition and consequences of intimate partner violence, the article reviews the research and related literatures to describe the (a) types of job interference tactics used by abusers, (b) employee-level consequences of partner violence, (c) victimized employee responses to intimate partner violence, (d) organizational-level consequences of partner violence, and (e) employer responses to intimate partner violence. Future research directions and workplace implications are discussed. PMID- 16217119 TI - The effectiveness of sexual assault nurse examiner (SANE) programs: a review of psychological, medical, legal, and community outcomes. AB - In sexual assault nurse examiner (SANE) programs, specially trained forensic nurses provide 24-hour-a-day, first-response medical care and crisis intervention to rape survivors in either hospitals or clinic settings. This article reviews the empirical literature regarding the effectiveness of SANE programs in five domains:(a) promoting the psychological recovery of survivors, (b) providing comprehensive and consistent post-rape medical care (e.g., emergency contraception, sexually transmitted disease [STD] prophylaxis), (c) documenting the forensic evidence of the crime completely and accurately, (d) improving the prosecution of sexual assault cases by providing better forensics and expert testimony, and (e) creating community change by bringing multiple service providers together to provide comprehensive care to rape survivors. Preliminary evidence suggests that SANE programs are effective in all domains, but such conclusions are tentative because most published studies have not included adequate methodological controls to rigorously test the effectiveness of SANE programs. Implications for practice and future research are discussed. PMID- 16217120 TI - Responsivity issues in the treatment of sexual offenders. AB - Andrews and Bonta (2003) have presented a model of effective correctional programming focusing on risk, need, and responsivity factors for a general criminal population. When applied to sexual offenders, the first two issues (risk assessment and treatment targets such as cognitive distortions) have received a great deal of research attention. However, little attention has been paid to responsivity issues: those factors that influence the extent to which clients benefit, or fail to benefit, from treatment programs. This article presents a review of several potential responsivity factors related to sexual offender treatment and outlines areas that require further research attention. PMID- 16217122 TI - The rhythm and blues of gene expression in the rodent pineal gland. AB - In all vertebrates, melatonin is rhythmically synthesized in the pineal gland and functions as a hormonal message, encoding for the duration of night. In rodents, the nocturnal rise and fall of the arylalkylamine N-ace-tyltransferase (AA-NAT) activity controls the rhythmic synthesis of melatonin. This rhythm is centered around the transcriptional regulation of the AA-NAT by two norepinephrine inducible transcription factors, the activator CREB (Ca2+/cAMP-response element binding protein) and the inhibitor ICER (inducible cAMP early repressor). CREB is activated by phosphorylation, which is one of the fastest responses in pinealocytes upon adrenergic stimulation, occurring within minutes. ICER in turn accumulates only after several hours, a time gap resulting from the required de novo protein synthesis upon adrenergic stimulation. However, these molecular components of neuroendocrine signaling in the rodent pineal gland are supplemented by the impact of a variety of neurotransmitters and neuromodulators, and by translational and post-translational mechanisms. By molecular crosstalk, those different inputs on pinealocytes seem to fine-tune the shape of the melatonin signal, by interacting at various levels with the NE/cAMP/pCREB/ICER pathway. In addition, these alternate signaling routes may be important in acute "emergency" situations. Together, concerted signaling events in the rodent pineal gland help to generate a stable and reliable hormonal message of darkness for the body, that, however, can be altered rapidly upon sudden and unexpected "error" signals. PMID- 16217124 TI - Measurement of melatonin and its metabolites: importance for the evaluation of their biological roles. AB - Many physiologic changes related to light-dark cycles and antioxidant effects have been related to melatonin (N-acetyl-5-methoxytryptamine) and its metabolites, N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK) and N1-acetyl-5 methoxykynuramine (AMK). In this review, we discuss some methodologies, in particular, those employing high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) assays to quantitatively determine melatonin, AFMK, and AMK. These approaches offer a highly specific and an accurate quantification of melatonin and its metabolites. These characteristics are essential to point out correctly the biological effects of these compounds in physiological and pathological conditions. PMID- 16217123 TI - Functional MT1 and MT2 melatonin receptors in mammals. AB - Melatonin, dubbed the hormone of darkness, is known to regulate a wide variety of physiological processes in mammals. This review describes well-defined functional responses mediated through activation of high-affinity MT1 and MT2 G protein coupled receptors viewed as potential targets for drug discovery. MT1 melatonin receptors modulate neuronal firing, arterial vasocon-striction, cell proliferation in cancer cells, and reproductive and metabolic functions. Activation of MT2 melatonin receptors phase shift circadian rhythms of neuronal firing in the suprachiasmatic nucleus, inhibit dopamine release in retina, induce vasodilation and inhibition of leukocyte rolling in arterial beds, and enhance immune responses. The melatonin-mediated responses elicited by activation of MT1 and MT2 native melatonin receptors are dependent on circadian time, duration and mode of exposure to endogenous or exogenous melatonin, and functional receptor sensitivity. Together, these studies underscore the importance of carefully linking each melatonin receptor type to specific functional responses in target tissues to facilitate the design and development of novel therapeutic agent. PMID- 16217125 TI - Antioxidative protection by melatonin: multiplicity of mechanisms from radical detoxification to radical avoidance. AB - Melatonin has been shown to protect against oxidative stress in various, highly divergent experimental systems. There are many reasons for its remarkable protective potential. Signaling effects comprise the upregulation of antioxidant enzymes, such as superoxide dismutases, peroxidases, and enzymes of glutathione supply, down-regulation of prooxidant enzymes, such as nitric oxide synthases and lipoxygenases, and presumably also the control of quinone reductase 2. Other mechanisms are based on direct interactions with several reactive oxygen and nitrogen species. Among these reactions, the capacity of easily undergoing single electron transfer reactions is of particular importance. Electron donation by melatonin is not only an aspect of direct radical scavenging, but additionally represents the basis for formation of the protective metabolites AFMK (N1-ace-tyl N2-formyl-5-methoxykynuramine) and AMK (N1-acetyl-5-methoxykynuramine). Recent investigations on mitochondrial metabolism indicate that melatonin as well as AMK are capable of supporting the electron flux through the respiratory chain, of preventing the breakdown of the mitochondrial membrane potential, and of decreasing electron leakage, thereby reducing the formation of superoxide anions. Radical avoidance is a new line of investigation, which exceeds mitochondrial actions and also comprises antiexcitatory effects and contributions to the maintenance of internal circadian phase relationships. PMID- 16217126 TI - Metabolic effects of melatonin on oxidative stress and diabetes mellitus. AB - Melatonin, which is synthesized in the pineal gland and other tissues, has a variety of physiological, immunological, and biochemical functions. It is a direct scavenger of free radicals and has indirect antioxidant effects due to its stimulation of the expression and activity of antioxidative enzymes such as glutathione peroxidase, superoxide dismutase and catalase, and NO synthase, in mammalian cells. Melatonin also reduces serum lipid levels in mammalian species, and helps to prevent oxidative stress in diabetic subjects. Long-term melatonin administration to diabetic rats reduced their hyperlipidemia and hyperinsulinemia, and restored their altered ratios of polyunsaturated fatty acid in serum and tissues. It was recently reported that melatonin enhanced insulin receptor kinase and IRS-1 phosphorylation, suggesting the potential existence of signaling pathway cross-talk between melatonin and insulin. Because TNF-alpha has been shown to impair insulin action by suppressing insulin receptor-tyrosine kinase activity and its IRS-1 tyrosine phosphorylation in peripheral tissues such as skeletal muscle cells, it was speculated that melatonin might counteract TNF alpha-associated insulin resistance in type 2 diabetes. This review will focus on the physiological and metabolic effects of melatonin and highlight its potential use for the treatment of cholesterol/lipid and carbohydrate disorders. PMID- 16217127 TI - On the role of melatonin in skin physiology and pathology. AB - Melatonin has been experimentally implicated in skin functions such as hair growth cycling, fur pigmentation, and melanoma control, and melatonin receptors are expressed in several skin cells including normal and malignant keratinocytes, melanocytes, and fibroblasts. Melatonin is also able to suppress ultraviolet (UV) induced damage to skin cells and shows strong antioxidant activity in UV exposed cells. Moreover, we recently uncovered expression in the skin of the biochemical machinery involved in the sequential transformation of l-tryptophan to serotonin and melatonin. Existence of the biosynthetic pathway was confirmed by detection of the corresponding genes and proteins with actual demonstration of enzymatic activities for tryptophan hydroxylase, serotonin N-acetyl-transferase, and hydroxyindole-O-methyltransferase in extracts from skin and skin cells. Initial evidence for in vivo synthesis of melatonin and its metabolism was obtained in hamster skin organ culture and in one melanoma line. Therefore, we propose that melatonin (synthesized locally or delivered topically) could counteract or buffer external (environmental) or internal stresses to preserve the biological integrity of the organ and to maintain its home-ostasis. Furthermore, melatonin could have a role in protection against solar radiation or even in the management of skin diseases. PMID- 16217128 TI - Physiological ischemia/reperfusion phenomena and their relation to endogenous melatonin production: a hypothesis. AB - Ischemia/reperfusion is a frequently encountered phenomenon in organisms. Prolonged ischemia followed then by reperfusion results in severe oxidative injury in tissues and organs; however, some species can tolerate such events better than others. In nature, arousal from hibernation and resurfacing from diving causes animals to experience classic ischemia/reperfusion and, somehow, these animals cope well with the potential oxidative stress. It has been documented that during these physiological ischemia/reperfusion events, the activities of several antioxidant enzymes and the levels of some small-molecular weight antioxidants become elevated. For example, the potent small-molecular weight antioxidant melatonin often attains especially high levels during these physiological ischemia/reperfusion events including during arousal from hibernation or in the newborns during delivery. Highly elevated melatonin production during these physiological ischemia/reperfusion episodes exhibits several features. First, this high melatonin production is transient and fits well with the time schedule of the physiological ischemia/reperfusion period; therefore, it is not related to the normal endogenous melatonin rhythm. Yet, this transient peak protects the animals from destructive oxidative processes that occur during these transition periods. Second, these high levels of melatonin seem to derive from several organs since pinealectomy does not totally reduce circulating levels of this agent. Third, high melatonin production present at arousal from hibernation or in the newborns at birth does not appear to be controlled by light, i.e., it occurs both during the day and at night, and the amplitudes of elevated melatonin levels are equivalent at these times. The significance of these findings is discussed herein. Based on currently available data, we hypothesize that melatonin plays an important role in the physiological ischemia/reperfusion, i.e., as a member of antioxidant defense system, to protect against the potential oxidative injury induced by the physiological ischemia/reperfusion. PMID- 16217129 TI - Melatonin and nitric oxide: two required antagonists for mitochondrial homeostasis. AB - The presence of nitric oxide (NO* ) in the mitochondria led to analysis of its source and functions in mitochondrial homeostasis. Studies have revealed the existence of a mtNOS isoform with similar features to nNOS, with some post traslational modifications, although without the typical signal peptide responsible for addressing proteins to mitochondrion. This isoform may account for the physiological production of NO* related to the respiratory control. During inflammatory conditions there is an excess of NO* in the mitochondria responsible for an increase in reactive oxygen and nitrogen species in sufficient amounts to compromise mitochondrial function. These conditions led to the discovery of the presence of an inducible mtNOS isoform with kinetic properties similar to iNOS. Experiments with knockout mice lacking either nNOS or iNOS further confirmed the existence of these two mtNOS isoforms in mitochondria. Although the increase in NO* in sepsis by inducible mtNOS may have important regulatory functions including the redistribution of oxygen into other pathways under hypoxia, it causes the production of excess NO* that is deleterious for the cell. Melatonin, an endogenous antioxidant, regulates mitochondrial respiration and bioenergetics and protects mitochondria from excess NO* by controlling the activity of mtNOS. PMID- 16217130 TI - Melatonin and Parkinson's disease. AB - Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzheimer's disease. It is characterized by a progressive loss of dopamine in the substantia nigra and striatum. However, over 70% of dopaminergic neuronal death occurs before the first symptoms appear, which makes either early diagnosis or effective treatments extremely difficult. Only symptomatic therapies have been used, including levodopa (l-dopa), to restore dopamine content; however, the use of l-dopa leads to some long-term pro-oxidant damage. In addition to a few specific mutations, oxidative stress and generation of free radicals from both mitochondrial impairment and dopamine metabolism are considered to play critical roles in PD etiology. Thus, the use of antioxidants as an important co-treatment with traditional therapies for PD has been suggested. Melatonin, or N-acetyl-5 methoxy-tryptamine, an indole mainly produced in the pineal gland, has been shown to have potent endogenous antioxidant actions. Because neurodegenerative disorders are mainly caused by oxidative damage, melatonin has been tested successfully in both in vivo and in vitro models of PD. The present review provides an up-to-date account of the findings and mechanisms involved in neuroprotection of melatonin in PD. PMID- 16217131 TI - Putting cancer to sleep at night: the neuroendocrine/circadian melatonin signal. AB - Physiological and pharmacological blood concentrations of melatonin inhibit tumorigenesis in a variety of in vivo and in vitro experimental models of neoplasia. Evidence indicates that melatonin's anticancer effects are exerted via inhibition of cell proliferation and a stimulation of differentiation and apoptosis. A new mechanism by which physiological and pharmacological blood levels of melatonin inhibit cancer growth in vivois via a melatonin-induced suppression of tumor linoleic acid (LA) uptake and its metabolism to the important mitogenic signaling molecule 13-hydroxyoctadecadienoic acid (13-HODE). Melatonin suppresses cAMP formation and inhibits tumor uptake of LA and its metabolism to 13-HODE via a melatonin receptor-mediated mechanism in both tissue isolated rat hepatoma 7288 CTC and human breast cancer xenografts. It has been postulated that in industrialized societies, light at night, by suppressing melatonin production, poses a new risk for the development of breast cancer and, perhaps, other cancers as well. In support of this hypothesis, light during darkness suppresses nocturnal melatonin production and stimulates the LA metabolism and growth of rat hepatoma and human breast cancer xenografts. Nocturnal dietary supplementation with melatonin, at levels contained in a melatonin-rich diet, inhibits rat hepatoma growth via the mechanisms described above. The nocturnal melatonin signal organizes tumor metabolism and growth within circadian time structure that can be further reinforced by appropriately timed melatonin supplementation. Dietary melatonin supplementation working in concert with the endogenous melatonin signal has the potential to be a new preventive/therapeutic strategy to optimize the host/cancer balance in favor of host survival and quality of life. PMID- 16217134 TI - Zinc content in lymphocytes and the activity of zinc ion efflux from lymphocytes in primary arterial hypertension. AB - Clinical observations and experimental data show that zinc (Zn) plays a role in regulating arterial blood pressure and in arterial hypertension etiopathogenesis. To determine the direction of changes in Zn metabolism in primary arterial hypertension, Zn absorption from the alimentary tract, Zn levels in blood serum, its content in lymphocytes, Zn efflux rate constants from lymphocytes, and urinary Zn excretion in patients with hypertension and in healthy subjects were studied. In this article, Zn levels in blood serum, its content in lymphocytes, and Zn efflux rate constants from lymphocytes are presented. In primary arterial hypertension, on the basis of this study, decreasing Zn levels in blood serum and its decreasing content in lymphocytes were found. The Zn efflux rate constants from lymphocytes increased at the initial stage of hypertension (mild arterial hypertension) and decreased in the late stage of the hypertension disease (severe arterial hypertension). Taking into consideration all of the directions of changes and the fact that Zn can be a factor that increases arterial blood pressure, the changes in Zn distribution can be regarded as having, to a certain extent, a protective character leading to weakening of the pressor reaction, assuming a genetic existence of relative or absolute Zn excess in the body. The changes of Zn distribution can lead, after some time, to Zn deficiency and the resulting metabolic changes (e.g., carbohydrate intolerance). PMID- 16217133 TI - Melatonin, aging, and age-related diseases: perspectives for prevention, intervention, and therapy. AB - The high incidence of age-related diseases in the increasing population of elderly people has stimulated interest in the search for protective agents that have the capability of preventing premature aging and delaying the onset of degenerative disorders. To preserve health in old age becomes a primary goal for biomedicine, because the increasing longevity in our societies is associated with a rise in morbidity. The difficulties in finding new approaches and safe strategies for prevention, intervention, and treatment are related to the lack of theoretical background as well as to insufficient models to test the efficacy of anti-aging agents. Melatonin is a prime candidate for slowing the aging process and targeting its underlying pathology. Melatonin has profound gerontoprotective and antioxidant activities. Because enhanced oxidative stress plays a crucial role in the aging process and chronic diseases associated with senescence, the adminstration of a potent amphiphilic antioxidant agent with high bioavailability such as melatonin may become a promising, safe, and effective intervention strategy to slow aging and the initiation and progression of age-related disorders. Investigations on melatonin and its anti-aging activity may be of great benefit in increasing life quality of the elderly. PMID- 16217135 TI - Effect of praseodymium(III) on zinc(II) species in human interstitial fluid. AB - A multiphase model of metal ion species in human interstitial fluid was constructed under physiological conditions. The effect of Pr(III) on Zn(II) species was studied. At the normal conditions, Zn(II) species mainly distribute in [Zn(HSA)], [Zn(IgG)], and [Zn(Cys)(2)H](+). With the Pr(III) level increased, the apparent competition of Pr(III) for ligands lead to the redistribution of Zn(II) species. PMID- 16217136 TI - Profiles of trace elements in toenails of Arab-Americans in the Detroit area, Michigan. AB - Exposure to environmental contaminants is complicated by factors related to socioeconomic status, diet, and other culturally conditioned risk behaviors. Determination of a trace element profile in toenails can be used as a tool in biomonitoring the exposure history or assessing the deficiency of a particular element in a study population, which can lead to a better understanding of environmental and disease risks. Toenail clippings from 259 Arab Americans (163 adults, 96 children) residing in a highly industrialized area were analyzed for Al, V, Cr, Mn, Co, Ni, Cu, As, Se, Mo, Cd, Ba, Tl, and Pb using an inductively coupled plasma-mass spectrometer. Mean concentrations were compared with published values, and the influence of age, gender, and other demographic factors were explored. Elevated levels of Ni in this population warrant further investigation. Significant differences in the mean concentration of Al, V, Cr, Mn, Cd, Pb, and Se exist between toenails of adults and children. Pearson correlation coefficients reveal strong significant associations among Cd, Cr, and Tl (p<0.05), in addition to other elements. These investigations provide insight into exposures and factors influencing exposures in this population while adding to the growing fund of knowledge surrounding use of toenails as a marker of exposure. PMID- 16217137 TI - In vitro evaluation of the antioxidant activity of calcium fructoborate. AB - Although increasing evidence shows the nutritional benefits of calcium fructoborate (CF) on animals and humans, its action mechanism has not been clearly identified. The present study aims to investigate the possible antioxidant function of CF. Based on its efficiency in skin wound healing, the authors tested whether CF possesses antioxidant properties on human keratinocytes cultures, in a complete serum-free medium (KMK-2; Sigma). The cells treated with CF (0-450 nmol/culture medium) were exposed to exogenous 100 micromol of hydrogen peroxide to mimic the oxidative stress. The changes in general cell oxidant production evaluated with dihydrorhodamine-123 showed that the intracellular reactive oxygen species (ROS) were markedly reduced by preincubation with CF. The maximum antioxidant activity was noticed at 90 nmol CF. To assess the reactivity of CF on ROS, we analyzed its ability to inhibit the superoxide- dependent auto oxidation of pyrogallol. The CF inhibited the pyrogallol auto-oxidation depending on time and concentration, which suggests its possible role as a superoxide radical scavenger. Taken together, our results indicate that CF has antioxidant activity, which could have clinical significance in protecting cells from oxidant induced injury. A hypothetic mechanism for the antioxidant activity of CF is proposed. PMID- 16217138 TI - Twenty-eight element concentrations in mane hair samples of adult riding horses determined by particle-induced X-ray emission. AB - The concentrations of 28 elements (Al, Br, Ca, Cl, Co, Cu, Cr, Fe, Ga, Hg, K, Mg, Mn, Mo, Na, Nb, Ni, P, Pb, Rb, S, Se, Si, Sr, Ti, V, Y, and Zn) were measured in mane hair by the particle-induced X-ray emission method. Except for Br, Cl, K, S, and P, the trace element concentrations in mane hair of horses are similar to literature values for human hair. The values obtained are not dependent on the horse's age, breed, and sex and could be used as reference values in the assessment of diseases and nutritional status in equines. PMID- 16217139 TI - Effect of excessive dietary fluoride on nutrient digestibility and retention of iron, copper, zinc, and manganese in growing pigs. AB - Ninety-six crossbred growing pigs were used to evaluate the effects of fluoride levels on growth performance, nutrient digestibility, and the retention of minerals in tissues. Four dietary treatments were formulated by supplementing fluorine (as NaF) to a corn-soybean basal diet (39.75 mg/kg F) to provide the following added fluorine levels: 0, 50, 100, and 150 mg/kg fluorine. The results showed pigs of the 100 and 150 mg/kg fluorine-added groups had decreased average daily gain (ADG) and increased feed gain ratio (F/G) compared to the control (p < 0.05). Apparent digestibility of protein and calcium in 100 and 150 mg/kg fluorine-treated groups was significantly lower than that of the control (p < 0.05). On the other hand, iron, copper, zinc, and manganese levels in most tissues of the 100 and 150 mg/kg fluorine groups were markedly changed compared to the control (p < 0.05). However, growth performance, nutrient digestibility, and mineral concentrations in all tissues of pigs were not significantly affected by the addition of 50 mg/kg fluorine (p > 0.05). Thus, this study suggested that excess fluoride levels could decrease growth performance and change the retention of iron, copper, zinc, and manganese in pigs. PMID- 16217140 TI - Effects of dietary zinc levels on the activities of enzymes, weights of organs, and the concentrations of zinc and copper in growing rats. AB - Zinc (Zn) is an essential nutrient that is required in humans and animals for many physiological functions, including immune and antioxidant function, growth, and reproduction. The present study was performed to investigate the effects of three Zn levels, including Zn adequate (35.94 mg/kg, as a control), Zn deficiency (3.15 mg/kg), and Zn overload (347.50 mg/kg) in growing male rats for 6 wk. This allowed for evaluation of the effects that these Zn levels might have on body weight, organ weight, enzymes activities, and tissues concentrations of Zn and Cu. The results showed that Zn deficiency has negative effects on growth, organ weight, and biological parameters such as alkaline phosphatase (ALP) and Cu-Zn superoxide dismutase (Cu-Zn SOD) activities, whereas Zn overload played an effective role in promoting growth, improving the developments of organs and enhancing immune system. Hepatic metallothionein (MT) concentration showed an identical increase tendency in rats fed both Zn-deficient and Zn-overload diets. The actual mechanism of reduction of Cu concentration of jejunum in rats fed a Zn overload diet might involve the modulation or inhibition of a Cu transporter protein by Zn and not by the induction of MT. PMID- 16217132 TI - A review of the multiple actions of melatonin on the immune system. AB - This review summarizes the numerous observations published in recent years which have shown that one of the most significant of melatonin's pleiotropic effects is the regulation of the immune system. The overview summarizes the immune effects of pinealectomy and the association between rhythmic melatonin production and adjustments in the immune system as markers of melatonin's immunomodulatory actions. The effects of both in vivo and in vitromelatonin administration on non specific, humoral, and cellular immune responses as well as on cellular proliferation and immune mediator production are presented. One of the main features that distinguishes melatonin from the classical hormones is its synthesis by a number of non-endocrine extrapineal organs, including the immune system. Herein, we summarize the presence of immune system-synthesized melatonin, its direct immunomodulatory effects on cytokine production, and its masking effects on exogenous melatonin action. The mechanisms of action of melatonin in the immune system are also discussed, focusing attention on the presence of membrane and nuclear receptors and the characterization of several physiological roles mediated by some receptor analogs in immune cells. The review focuses on melatonin's actions in several immune pathologies including infection, inflammation, and autoimmunity together with the relation between melatonin, immunity, and cancer. PMID- 16217141 TI - Effect of selenite treatment on ultrastructural changes in experimental diabetic rat bones. AB - It is known that streptozotocin (STZ)-induced diabetes causes functional and structural alterations in some types of tissue and organ. A number of methods have been used to characterize the properties of diabetic tissues and their diagnosis. Selenium compounds, playing an antioxidant role, can restore some altered metabolic parameters and diminished functions in experimental diabetes. The first aim of the present study was to investigate the effects of STZ-induced diabetes on structural properties of rat long bones. Electron and light microscopic observations showed deleterious alterations in the structure of the diabetic rat long bones, the most prominent effect being in osteocytic cells. Fine cytoplasmic processes of the osteocytes seemed to be shortened, and diabetes affected the normal cytoplasmic processes in a negative manner. The second aim of the present study was to evaluate the effects of sodium selenite treatment for 4 wk on the long bones of the diabetic rats. Electron and light microscopic observations demonstrated that sodium selenite treatment prevented the STZ induced structural as well as ultrastructural changes in the long bones of the rats. In conclusion, this study first showed that a period of 5-wk diabetes was enough to cause some important and degenerative changes in the structure of the bone tissues, and, second, it demonstrated that sodium selenite treatment of the diabetic rats could normalize these alterations. PMID- 16217142 TI - Evaluation of Nigerian animal feeds by particle-induced X-ray emission. AB - There is need to evaluate the locally available animal feeds in Nigeria so as to be able to combine them in acceptable proportions to the animals to achieve the desired growth rate. The technique of particle-induced X-ray emission (PIXE) was employed for the evaluation of these locally available animal feeds, which include Panicum maximum (Guinea grass), Cynodon plectostachyum (grass), Leucaena leucephala (legume), Calopogonium mucunoides (legume), Gliricidia sepium (legume), Euphorbia polychrome (legume), Pueraria phaseloides (legume), and Centrosema pubescens (legume). The proton beam delivered by the 2.5-MV AN 2,000 Van de Graaff accelerator at the International Centre for Theoretical Physics Laboratori Nazionali di Legnaro (LNL), Padova, Italy was used for the PIXE measurements. Twenty-one different elements were detected at various concentrations and their nutritional effects on different animals are discussed. PMID- 16217143 TI - Halothane sensitizes the guinea-pig heart to pharmacological IKr blockade: comparison with urethane anesthesia. AB - Potential utility of halothane-anesthetized guinea pigs for detecting drug induced repolarization delay was analyzed in comparison with urethane-anesthesia (n = 4 for both groups). Basal QT interval was significantly greater under halothane-anesthesia than urethane-anesthesia (192 +/- 7 vs 132 +/- 5 ms, respectively), whereas the reverse was true for the heart rate (190 +/- 7 vs 248 +/- 11 beats/min, respectively). The typical I(Kr)-blocker dl-sotalol (0.1 to 3 mg/kg, i.v.) induced dose-related bradycardia and QT interval prolongation under each anesthesia. The extent of maximum prolongation in the QT interval was greater under halothane-anesthesia than urethane-anesthesia (+101 +/- 15 vs +49 +/- 3 ms, respectively), whereas that of peak change in the heart rate was smaller under the former than the latter (-49 +/- 8 vs -63 +/- 5 beats/min, respectively). Pretreatment of the animals under urethane-anesthesia with the selective I(Ks) blocker chromanol 293B (n = 6) increased the extent of the dl sotalol-induced QT interval prolongation to +57 +/- 8 ms, which was only 0.56 times of that under the halothane-anesthesia, whereas the pretreatment increased the peak change in the heart rate to -76 +/- 12 ms. These results indicate that the halothane-anesthesia may effectively sensitize the guinea-pig heart to pharmacological I(Kr) blockade. PMID- 16217144 TI - Inhibitory effect of thiopental on ultra-rapid delayed rectifier K+ current in H9c2 cells. AB - Using the whole-cell voltage clamp technique, we investigated the effects of thiopental on membrane currents in H9c2 cells, a cell line derived from embryonic rat heart. Thiopental blocked a rapidly activating, very slowly-inactivating ultra-rapid type I(Kur)-like outward K(+) current in a concentration-dependent manner. The half-maximal concentration (IC(50)) of thiopental was 97 microM with a Hill coefficient of 1.2. The thiopental-sensitive current was also blocked by high concentrations of nifedipine (IC(50) = 9.1 microM) and 100 microM chromanol 293B, a blocker of slowly activating delayed rectifier K+ current (I(Ks)), but was insensitive to E-4031, an inhibitor of rapidly activating delayed rectifier K(+) current (I(Kr)). TEA (tetraethylammonium) at 5 mM and 4-AP (4-aminopiridine) at 1 mM reduced the K(+) current to 30.8 +/- 12.2% and 20.5 +/- 6.5% of the control, respectively. Using RT-PCR, we detected mRNAs of Kv2.1, Kv3.4, Kv4.1, and Kv4.3 in H9c2 cells. Among those, Kv2.1 and Kv3.4 have I(Kur)-type kinetics and are therefore candidates for thiopental-sensitive K(+) channels in H9c2 cells. This is the first report showing that thiopental inhibits I(Kur). This effect of thiopental may be involved in its reported prolongation of cardiac action potentials. PMID- 16217151 TI - Compression therapy for venous ulcers. PMID- 16217145 TI - Regulated expression and function of the somatodendritic catecholamine neurotransmitter transporters. AB - Termination of neurotransmission at catecholaminergic synapses is well documented by the transporters for dopamine and norepinephrine, members of the Na(+)/Cl(-) dependent neurotransmitter transporter family, which accumulates released transmitters within their nerve endings, respectively. Although somatodendritic expression of the transporters and the effects of cocaine and amphetamine on those have been reported, their role is still obscure. Recent findings of the transporter function as an ion channel and/or its reverse transport property provide a clue to identify the role of these transporters in the somatodendrites and their consequential interaction with uptake inhibitors. Differences in ionic environment and maturity of the release machinery in the somatodendrites at developmental stages influence the transporter functions, resulting in the formation of both positive and negative feedback loop of catecholaminergic neurons. PMID- 16217152 TI - Purchasing responsibilities for surgical dressings. PMID- 16217153 TI - Moving toward consensus on deep tissue injury and pressure ulcer staging. PMID- 16217154 TI - Skin care protocols for pressure ulcers and incontinence in long-term care: a quasi-experimental study. AB - OBJECTIVE: To evaluate the effect on pressure ulcer prevalence, incidence, and healing time of incorporating use of a specific body wash and a skin protectant into skin care protocols that are based on guidelines from the Agency for Health Care Policy and Research. DESIGN: Quasi-experimental intervention study. SETTING: 2 rural long-term-care facilities. PARTICIPANTS: A convenience sample of 136 residents at 2 rural long-term-care facilities during a 3-month preintervention and a 3-month postintervention period. INTERVENTIONS: A 3-month preintervention observation period (baseline) was followed by a staff in-service session, in which the use of a body wash and a skin protectant was introduced into skin care protocols, and a 3-month postintervention observation period. The skin care protocols included skin assessment techniques, prevention and treatment strategies for Stage I and Stage II pressure ulcers, and management of incontinence. MAIN OUTCOME MEASURES: Differences in the occurrence and healing time of Stage I and Stage II pressure ulcers before and after introduction of use of a body wash and a skin protectant into skin care protocols and the occurrence rate of urinary and fecal incontinence. RESULTS: Stage I and Stage II pressure ulcers significantly decreased from 35 preintervention to 14 postintervention (t = 19.48, df = 47, P = .05). The prevalence of pressure ulcers preintervention was 11.3%, compared with 4.8% postintervention (t = 2.47, df = 1.0, P = .24), The change in the incidence of pressure ulcers was significant (t = 8.48, df = -2.0, P = .01), with 32.7% preintervention and 8.9% postintervention. Healing time for pressure ulcers ranged from 4 to 70 days preintervention (mean [M] = 22.72 +/- 18.25) to 6 to 49 days postintervention (M = 16.0 +/- 12.93). The decrease in pressure ulcer healing time (rapid, medium, and long) preintervention to postintervention was statistically significant (chi 2= 14.9, P = .001). The presence of fecal and urinary incontinence was significantly associated with the development of Stage I and Stage II pressure ulcers (chi2= 44.8, P = .000). CONCLUSIONS: Implementation of skin care protocols that included use of a body wash and a skin protectant reduced the incidence of Stage I and Stage II pressure ulcers and decreased healing time. The skin protectant and body wash used in the protocols were found to be effective in preventing and treating Stage I and Stage II pressure ulcers. PMID- 16217155 TI - Maggot debridement therapy: free-range or contained? An in-vivo study. AB - OBJECTIVE: To determine which method of maggot debridement therapy--free-range or contained--is more effective for wound healing. METHODS: In vivo study of 64 patients with 69 chronic wounds that showed signs of gangrenous or necrotic tissue. Patients were treated with either free-range or contained maggot debridement therapy according to maggot availability, dressing difficulty, and physician preference. RESULTS: Significantly better outcomes were achieved with the free-range technique versus the contained technique (P = .028). With the free range technique, the mean number of maggot applications and the total number of maggots per treatment were significantly lower than with the contained application technique (P = .028 and P < .001, respectively). CONCLUSION: This clinical in vivo study supports in vitro studies in which containment of maggots was found to reduce the effectiveness of maggot debridement therapy. PMID- 16217156 TI - Management of vascular leg ulcers. PMID- 16217159 TI - Current status of hematopoietic stem cell transplantation after nonmyeloablative conditioning. AB - PURPOSE OF REVIEW: Allogeneic hematopoietic stem cell transplantation was originally developed as a form of rescue from high-dose chemoradiotherapy, which is given both to eradicate malignancy and provide sufficient immunosuppression for allogeneic engraftment. However, it was observed that allogeneic immunocompetent cells transplanted with the stem cells, or arising from them, mediated therapeutic antitumor effects independent of the action of the high-dose therapy. This was termed a graft-versus-tumor effect. This has prompted the recent development of nonmyeloablative conditioning regimens for allogeneic hematopoietic stem cell transplantation that have opened the way to include elderly patients and those with comorbid conditions. RECENT FINDINGS: Recent retrospective studies comparing hematopoietic stem cell transplantation after myeloablative or nonmyeloablative regimens suggested that the use of nonmyeloablative conditioning might be associated with lower transplant-related toxicity, lower nonrelapse mortality, and at least similar intermediate-term progression-free survival. SUMMARY: Hematopoietic stem cell transplantation after nonmyeloablative conditioning might become the procedure of choice also for younger patients. Phase 3 studies are needed to determine outcomes for different diseases and age groups. PMID- 16217160 TI - The lymphocyte homing receptors: gatekeepers of the multistep paradigm. AB - PURPOSE OF REVIEW: Lymphocyte trafficking is regulated by adhesion molecules mediating the initial shear-resistant binding of circulating cells to target tissue endothelium (Step 1). This review focuses on the current and emerging perspectives of the biology of these 'homing receptors.' RECENT FINDINGS: The conventional multistep paradigm holds that leukocyte migration represents a cascade of events, initiated by tethering and rolling interactions of leukocytes on the endothelial surface (Step 1). These interactions are indispensable, required to dampen velocities sufficiently to allow cells to sense the local chemokines and/or other inflammatory signals resulting in activation of integrin adhesiveness (Step 2), with ensuing firm adherence on the vessel wall (Step 3) followed by endothelial transmigration (Step 4). Mechanistic studies now suggest that some effectors of Step 1 interactions themselves activate integrin adhesiveness and trigger transmigration--in some cases by forming complexes with integrins--thus bypassing the need for chemokine signaling. These findings force a reconsideration of the multistep paradigm, and shift focus now to identifying all relevant effectors of Step 1 interactions using adherence assays performed under shear stress to mimic the dynamic conditions of blood flow. SUMMARY: Recent findings suggest that homing receptors are not merely molecular brakes. The cross talk among the homing receptors and integrins opens a new 'avenue' to lymphocyte migration, suggesting that homing receptors may be sufficient alone, in some cases, to perform the function the name implies. PMID- 16217161 TI - Regulatory T cells: prospective for clinical application in hematopoietic stem cell transplantation. AB - PURPOSE OF REVIEW: Regulatory T cells exert a dominant effect in controlling autoimmunity and maintaining peripheral tolerance. Regulatory T cells are also involved in preventing allograft rejection and graft versus host disease. Cellular therapy with expanded regulatory T cells represents a promising approach to control T-cell mediated pathology. In this review we will summarize the efforts to design new methods for expanding regulatory T cells and exploit their regulatory function as cellular therapy for the treatment of graft versus host disease after hematopoietic stem cell transplantation. RECENT FINDINGS: Among CD4+ T cells, the best described are the naturally occurring CD4+CD25+ regulatory T cells and type 1 regulatory T cells. Recent progress has been made in the characterization of both subsets in terms of isolation and induction, respectively. However, a clear definition of their mechanisms of action has still to be achieved. SUMMARY: Better understanding of the mechanisms of suppression mediated by regulatory T cells might enable their use to modulate specific immune responses. Moreover, the recent development of methods allowing the ex-vivo expansion of regulatory T cells, to provide sufficient number of cells for in vivo infusion, represents the first step toward the use of these cells as cellular therapy for the treatment of immunologic and hematological diseases. PMID- 16217162 TI - Antithymocyte globulin for prevention of graft-versus-host disease. AB - PURPOSE OF REVIEW: This review aims to discuss current clinical trial using antithymocyte globulin in the context of hematopoietic stem cell transplantation. Published papers, as well as abstracts from European and American meetings, have been used together with original data. RECENT FINDINGS: The review shows that antithymocyte globulin is used frequently in Europe in the setting of unrelated or family mismatched donor transplants. This may not be the case for American centers, reluctant to introduce antithymocyte globulin possibly due to known side effects such as prolonged immune deficiency and increased incidence of infections, including life threatening Epstein-Barr virus-associated lymphoproliferative disorders. Results from few randomized trials show that antithymocyte globulin reduces acute and chronic graft-versus-host disease, and this is in keeping with results from retrospective analysis on large number of patients. SUMMARY: Several issues remain open, such as optimal dose, optimal timing with respect to infusion of stem cells and whether the commercially available agents have comparable effects. Prospective trials are needed to answer these important questions. PMID- 16217163 TI - Progress in hematopoietic stem cell transplantation in multiple myeloma. AB - PURPOSE OF REVIEW: An attempt is made to value the place of autologous and allogeneic hematopoietic stem cell transplantation in the treatment of multiple myeloma. Recent progress with each treatment modality has significantly changed the outcome for myeloma patients. RECENT FINDINGS: The gold standard for conditioning treatment before autologous transplantation is melphalan 200 mg/m, and the use of peripheral blood stem cells is superior to that of bone marrow. Tandem autologous transplantation is superior to single autologous transplantation, at least for patients who do not respond with complete remission to the first transplant. Allogeneic transplantation using standard high-dose myeloablative conditioning reduces the relapse rate in comparison with autologous transplantation; however, treatment-related mortality is significantly higher. Progress in allogeneic transplantation has been significant, and transplant related mortality has been reduced. The recent use of nonmyeloablative reduced intensity conditioning has significantly decreased transplant-related mortality; however, the relapse rate is higher than with standard conditioning, and therefore no significant improvement in overall survival has as yet been seen. Relapse following allogeneic transplantation can be counteracted by donor lymphocyte infusions, but the survival results are so far inconclusive. SUMMARY: Autologous transplantation is still the first choice for most myeloma patients, but it does not seem to be curative. Therefore, allogeneic transplantation should be continued in clinical trials exploring new conditioning methods, the graft versus-myeloma effect of new cell types, and its combination with new targeted drugs such as bortezomib and lenalidomide. PMID- 16217165 TI - Platelet gels and releasates. AB - PURPOSE OF REVIEW: This review addresses potential roles for platelets and their derivatives (gels, releasates, and lysates) as therapeutic agents for regenerative medicine. Recognizing that activated platelets release chemotactic and growth factors, investigators have attempted to enhance tissue regeneration by applying platelets and various derivatives directly into sites of surgical interventions or injuries. This review analyzes the physiologic basis for this approach to tissue healing and examines the knowledge that has been derived from recent and relevant reports of in-vitro and in-vivo studies. RECENT FINDINGS: In vitro studies have established that platelets and their derivatives accelerate proliferation of an array of cells involved in soft and bony tissue regeneration. These effects have been evaluated, also, in vivo in humans and in animals. The outcomes of in-vivo studies are considerable less homogeneous than the outcomes of in-vitro investigations. The resultant discrepancies reflect not only differences of technical protocols, but also the greater complexity of healing vital tissues compared with circumscribed in-vitro studies. SUMMARY: The preponderance of evidence indicates that platelets and their derivatives have the potential for a substantial therapeutic role in tissue regeneration. The results of recent research indicate that platelet-derived growth factors act in synergy with plasma-derived factors to activate a complex network of autocrine functions that modulate healing. Platelet-derivative products are promising therapeutics that offer new opportunities for research and applications of tissue engineering. PMID- 16217166 TI - Transfusion-related acute lung injury. AB - PURPOSE OF REVIEW: Transfusion-related acute lung injury is an uncommon complication of blood transfusion typically manifested by shortness of breath, fever, and hypotension. Transfusion-related acute lung injury is an important cause of transfusion-related morbidity and mortality. RECENT FINDINGS: Much about the pathogenesis, treatment, and prevention of transfusion-related acute lung injury is poorly understood or is controversial. There is increasing recognition that transfusion-related acute lung injury is an important clinical syndrome, causing most transfusion-related deaths. SUMMARY: In this report, what is known about transfusion-related acute lung injury is summarized with particular emphasis on recent studies. Some of the areas in which knowledge and/or consensus are currently lacking are identified. PMID- 16217167 TI - Massive blood transfusion for trauma. AB - PURPOSE OF REVIEW: Massive blood transfusion saves the lives of thousands of severely injured patients each year, but it does so in the context of the evolving epidemiology of injury, of trauma centers and trauma systems, and of blood safety and new technologies for hemorrhage control. This article reviews recent knowledge and advances that impact on the use and effectiveness of massive transfusion. RECENT FINDINGS: Injury is rapidly becoming the second leading cause of death in the world. These deaths are highly preventable with social and engineering controls and good trauma care. Massive transfusion is readily available, safe, effective, and cheap in the context of modern trauma center care. However, aged blood products can cause transfusion-related acute lung injury, and better blood storage systems are under development. Recent work has improved understanding of the coagulopathy associated with acidosis and provided guidance for limiting dilutional coagulopathy. Nevertheless, massive transfusion always leads to coagulopathy and so is at best an adjunct to good surgical care. Better drugs and devices for hemorrhage control, such as recombinant activated factor VII and hemostatic bandages, are in development. SUMMARY: Injury is a major public health and medical system problem. Progress in basic science, clinical care, and the development of better hemorrhage control devices are all improving outcome for massively transfused patients. Investment in trauma care and supporting blood supply systems is highly cost effective. PMID- 16217168 TI - Hereditary angioedema. AB - PURPOSE OF REVIEW: Hereditary angioedema is an autosomal-dominant deficiency of C1 inhibitor--a serpin inhibitor of kallikrein, C1r, C1s, factor XII, and plasmin. Quantitative or qualitative deficiency of C1 inhibitor leads to the generation of vasoactive mediators, most likely bradykinin. The clinical syndrome is repeated bouts of nonpruritic, nonpitting edema of the face, larynx, extermities, and intestinal viscera. Recently, investigators, physicians, and industry have demonstrated a renewed interest in the biology and treatment of hereditary angioedema. RECENT FINDINGS: Investigators have generated a C1INH-/- mouse model that has demonstrated the importance of the contact activation system for hereditary angioedema-related vascular permeability. An interactive database of mutations is available electronically. Investigators have continued exploration into mRNA/protein levels. The proceedings of a recent workshop have been impressive in the scope and depth. Clinicians have produced consensus documents and expert reviews. The pharmaceutical industry has initiated clinical trails with novel agents. SUMMARY: Hereditary angioedema is often misdiagnosed and poorly treated. Diagnosis requires careful medical and family history and the measurement of functional C1 inhibitor and C4 levels. Attenuated androgens, anti fibrinolytics, and C1 inhibitor concentrates are used for long-term and preprocedure prophylaxis, but have significant drawbacks. C1 inhibitor concentrates and fresh frozen plasma are available for acute intervention. The mainstays of supportive care are airway monitoring, pain relief, hydration, and control of nausea. New agents such as recombinant C1 inhibitor, kallikrein inhibitors, and bradykinin inhibitors may offer safer and more tolerable treatments. PMID- 16217169 TI - Pretransfusion trigger platelet counts and dose for prophylactic platelet transfusions. AB - PURPOSE OF REVIEW: To assess critically both the blood platelet counts that prompt a platelet transfusion (i.e. trigger) in various clinical settings in patients with thrombocytopenia caused by marrow failure and the dose of platelets infused (i.e. number per each transfusion) for optimal hemostasis, feasibility, and safety. RECENT FINDINGS: Definitive studies (e.g. well-designed, prospective, randomized clinical trials) are not available either historically or at present to support evidence-based decisions. Instead, retrospective reviews and anecdotal reports provide observational data to assist in best guess clinical practices. SUMMARY: Reasonable clinical practice, until more definitive data become available, is to transfuse enough platelets per each transfusion to maintain the blood platelet count >10 x 10/L in stable nonbleeding patients, >20 x 10(9)/L in unstable nonbleeding patients, and >50 x 10(9)/L in bleeding patients or in those undergoing invasive procedures. PMID- 16217171 TI - Bibliography current world literature. Hematopoietic stem cell transplantation. PMID- 16217170 TI - Seasonal blood shortages can be eliminated. AB - PURPOSE OF REVIEW: This review is designed to help readers understand seasonal blood shortages and provide solutions through the use of technology that can increase the number of red blood cell units collected and the use of recruitment and marketing initiatives that appeal to the increasingly diverse donor base. RECENT FINDINGS: Seasonal shortages are, in reality, mostly shortages of group O red blood cells and occur most commonly during midsummer and early winter. The shortages occur primarily from increased use of group O red blood cells at times of decreased donor availability. While reducing the disproportionate use of red cells will help, blood centers can more quickly reduce the seasonal deficits by using automated red cell technology to collect double red blood cell units; targeted marketing programs to provide effective messages; seasonal advertising campaigns; and recognition, benefits, and incentives to enhance the donor motivation donation threshold. A multi-level approach to increasing blood donations at difficult times of the year can ensure that donations are increased at a time when regular donor availability is decreased. SUMMARY: Seasonal blood shortages can be eliminated by understanding the nature of the shortages, why and when they occur, and using more sophisticated recruitment and marketing strategies as well as automated collection technologies to enhance the blood supply. PMID- 16217172 TI - Cytokines and normal sleep. AB - PURPOSE OF REVIEW: Cytokines are mediators of immune system responses with multiple biologic actions on several target tissues. Over the past two decades, research has explored the interactions between cytokines and sleep mechanisms of the brain. This short review highlights selected findings that have advanced our understanding of the relation between cytokines and sleep. RECENT FINDINGS: A complex network of cytokines and their receptors exists in brain. Cytokines may either promote or inhibit sleep. Of cytokines studied thus far, evidence indicates that interleukin-1 and tumor necrosis factor play a role in the regulation of non-rapid eye movement sleep. Their sites of action for regulating such sleep likely include the hypothalamic preoptic area and the basal forebrain. Mechanisms of action include direct receptor-mediated effects on neurons and the synthesis and release of numerous transmitters, peptides, and hormones that lead to subsequent changes in sleep. Among others, the cascade of responses induced by cytokines that may lead to subsequent alterations in sleep includes alterations in nitric oxide synthesis and effects on neurohormonal systems such as growth hormone releasing hormone. The activation by cytokines of the hypothalamic pituitary-adrenal axis also influences sleep. Studies suggest that there is a significant overlap between neurohormonal systems such as the somatotropic and hypothalamic-pituitary-adrenal axes and cytokines, particularly with regard to their effects on sleep-wake regulation. SUMMARY: There is increasing evidence of a role for cytokines in regulating spontaneous non-rapid eye movement sleep. The somatotropic hormonal system and hypothalamic-pituitary-adrenal axis mediate, in part, the effects of cytokines on sleep. PMID- 16217173 TI - Recognition and management of complex sleep-disordered breathing. AB - PURPOSE OF REVIEW: The recent rapid evolution of our understanding of the mechanisms involved in control of respiration during sleep has yielded new insights to guide our care of difficult-to-treat sleep apnea patients with complex sleep-disordered breathing. This review will describe these recent advances in the literature and suggest a model for their incorporation into clinical practice. RECENT FINDINGS: Control of respiration during sleep shows amplified instability relative to that seen during wake in these difficult patients. Baseline (eupneic) carbon dioxide levels as well as the responsiveness of the ventilatory system to changes in carbon dioxide are all-important in this relative instability. Furthermore, the instability seen during sleep varies widely across sleep states. A further refinement of our definition of stable and unstable sleep has been developed that directly informs our understanding of the control of respiration across a night of sleep. SUMMARY: Complex sleep-disordered breathing is a distinct form of sleep apnea. It has recognizable characteristics that are present without, and often worsened during, positive airway pressure treatment. Both sleep state stability and the behavior of the respiratory control system contribute to this complexity. It is only with a clear understanding of the factors contributing to complex sleep-disordered breathing that implementation of truly effective clinical therapy can be achieved for this disorder, which to date is poorly controlled. PMID- 16217174 TI - Neurobehavioral effects of obstructive sleep apnea: an overview and heuristic model. AB - PURPOSE OF REVIEW: Obstructive sleep apnea can cause significant daytime behavioral and adaptive deficits, conventionally called 'neurobehavioral' because they are presumed to be mediated by the brain. The past few years have witnessed a marked increase in research into the neurobehavioral effects of obstructive sleep apnea, making it difficult for researchers and clinicians to stay abreast of the field. This article summarizes recent findings on the neurobehavioral effects of adult and pediatric obstructive sleep apnea, and presents a heuristic model to guide future research and clinical conceptualizations. RECENT FINDINGS: Recent publications have suggested overlapping but distinct neurobehavioral deficits that tend to accompany obstructive sleep apnea in the adult versus childhood years. There have been exciting new developments into the mechanisms by which obstructive sleep apnea may result in these deficits, including findings based upon advanced neuroimaging tools and carefully controlled animal research. There has also been accumulating evidence for potential moderators of morbidity; that is, factors that alter the nature or severity of neurobehavioral deficit resulting from obstructive sleep apnea. Task- and context-related factors that may affect outcome have been identified, as have potential markers for individual risk and resiliency. SUMMARY: The relation between obstructive sleep apnea and neurobehavioral deficit is probably not dependent on a single mediating mechanism, nor is it invariant across individuals. In outlining several potential moderating factors, the model presented here was designed to provide a guide for future research and a way of thinking about obstructive sleep apnea that better captures the wide variation in neurobehavioral outcome seen by practicing clinicians. PMID- 16217175 TI - Sleep-disordered breathing and cardiovascular health. AB - PURPOSE OF REVIEW: Sleep-disordered breathing is a widely prevalent condition and may have serious medical, social, and economic consequences. This review evaluates the role of sleep-disordered breathing in cardiovascular morbidity and mortality on the basis of recent literature. RECENT FINDINGS: Epidemiologic studies, retrospective reviews, and prospective clinical trials suggest a strong association between sleep-disordered breathing and adverse cardiovascular events. Individuals with sleep-disordered breathing have a higher prevalence of hypertension, which attenuates with treatment. Furthermore, the presence of sleep disordered breathing augurs an increased risk of coronary artery disease, cardiac arrhythmias, and heart failure. SUMMARY: Recent research provides an emerging evidence of the role of sleep-disordered breathing as a risk factor for diverse cardiovascular disorders. PMID- 16217176 TI - Sleep and the medical profession. AB - PURPOSE OF REVIEW: This review addresses the way in which sleep physiology influences the medical profession and health care delivery. RECENT FINDINGS: The training process for the medical professional has undergone dramatic changes over the past century. In recent times, however, the complexity and level of care delivered has out-stripped a trainee's ability to forego sleep and is compromising both physician and patient safety and thereby threatens the foundation of the profession. Recently, significant strides have been made in our understanding of sleep loss and consequences to physicians-in-training. Nevertheless, the implementation of changes fostered by such findings faces numerous conceptual and practical obstacles. This review updates the reader on recent evidence for changing the way medical professionals are trained, and opines on how solutions generated from such research should be embraced. Additionally, the deficiencies in our current understanding of sleep and medical training are identified so that future research can be undertaken in such areas. SUMMARY: Acknowledging the defects in our current system of training physicians and enacting further changes is sorely needed to improve patient safety and the well-being of physicians-in-training. PMID- 16217177 TI - Manifestations of cystic fibrosis diagnosed in adulthood. AB - PURPOSE OF REVIEW: This review highlights the phenotypic features that lead to the diagnosis of cystic fibrosis in adults, and the prognosis of these patients. RECENT FINDINGS: With the widespread availability of genetic testing and a greater appreciation of the clinical spectrum of the disease, the diagnosis of cystic fibrosis is being made with increasing frequency in adults. Clinical features that lead to the diagnosis include respiratory symptoms and chronic airway infection with typical cystic fibrosis pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus, as well as nontuberculous mycobacteria. Often these patients have previously received diagnoses of asthma, chronic bronchitis, or emphysema. Pancreatic insufficiency is much less common in the adult receiving the diagnosis, but pancreatitis occurs with greater frequency. Occasionally, individuals receive diagnoses of apparent single-organ manifestations such as idiopathic pancreatitis or congenital bilateral absence of the vas deferens, but with negligible involvement of the respiratory tract. On rare occasions, patients receiving the diagnosis as adults can present with classic features of the disease. Although lung disease is generally less severe in cystic fibrosis patients receiving the diagnosis as adults than in adult patients who received the diagnosis as infants, the extent of bronchiectasis can nonetheless be severe. The clinical course of patients receiving a diagnosis of cystic fibrosis in adulthood is largely unknown, but frequently they have milder disease and a more favorable prognosis. SUMMARY: Clinicians must be aware of the potential for adults with chronic respiratory tract infections, unexplained bronchiectasis, congenital bilateral absence of the vas deferens, or pancreatitis to have cystic fibrosis despite the age at presentation. PMID- 16217178 TI - Update on the role of exercise in cystic fibrosis. AB - PURPOSE OF REVIEW: The role of exercise in patients with cystic fibrosis has been recognized for as long as cystic fibrosis has been identified as a clinical syndrome, as exercise intolerance has always been a hallmark of disease progression. RECENT FINDINGS: Work published in the past year has shed new light on several aspects of this field, including physiologic responses to exercise, responses to exercise programs, and the epidemiologic and prognostic implications of activity and exercise testing. SUMMARY: Barker's work is perhaps the most compelling, as it highlights the overwhelming interest and belief in the utility of exercise testing and prescription among cystic fibrosis physicians and the contrasting paucity of programs that test their patients and prescribe exercise for them. More studies are needed to identify the form of exercise programs and tests that are most likely to be effective and to be used by cystic fibrosis centers and patients. PMID- 16217179 TI - Update on the evaluation of pancreatic exocrine status in cystic fibrosis. AB - PURPOSE OF REVIEW: Pancreatic functional status has a very strong effect on outcome in cystic fibrosis and pancreatic insufficiency requires lifelong treatment with pancreatic enzymes. Traditionally, clinical signs and symptoms have been used to decide who should be treated with pancreatic enzyme supplements; however, recent studies show that patients with cystic fibrosis are both undertreated and over-treated using this approach. This paper reviews recent progress in the development of noninvasive, indirect tests of pancreatic function for use as diagnostic tools for patients with cystic fibrosis. RECENT FINDINGS: Breath testing using C-labeled fat and measurement of several pancreatic enzymes in stool, such as chymotrypsin, lipase, and elastase have been explored as ways to define pancreatic functional status. Fecal elastase has good sensitivity, specificity, and positive and negative predictive value for defining severe pancreatic insufficiency in patients with cystic fibrosis and appears to be more useful than measurement of other fecal enzymes. Its role in milder pancreatic insufficiency and in disease states other than cystic fibrosis, such as chronic pancreatitis, is less clear. SUMMARY: Several new noninvasive, indirect tests of pancreatic function have been developed to aid in the definition of pancreatic functional status in patients with cystic fibrosis. An objective measure of pancreatic functional status should be obtained in all patients with cystic fibrosis, and the recent development of new screening tools such as fecal elastase makes this feasible. PMID- 16217181 TI - Early detection of lung disease in preschool children with cystic fibrosis. AB - PURPOSE OF REVIEW: Babies with cystic fibrosis are born with essentially normal lungs but rapidly develop inflammation and infection. Evaluation is generally based on history and physical examination until school age, when children become able to perform spirometry. In these 'silent years', however, unrecognized lung damage can occur. Recently, techniques to improve the evaluation of preschool children have been developed, and an assessment of where they fit into clinical and research practice is now needed. This is particularly urgent to select groups of children for new, phenotype-specific therapies, and monitor their effects. RECENT FINDINGS: The techniques in which there have been major recent advances include physiologic methods, particularly indices of gas mixing; imaging techniques, especially high-resolution computed tomographic scanning; and bronchoscopic studies. The clinical roles of these techniques need to be explored, as do comparisons between techniques. SUMMARY: The preschool years need no longer be silent. Some simple techniques such as spirometry can be performed in much younger children than conventionally believed to be practical. Others currently require sophisticated apparatus (lung clearance index) or carry potential risks (radiation, high-resolution computed tomographic scanning). We need good long-term, prospective comparisons of these techniques, with measurement of inflammatory markers. We also need to know which are sufficiently reproducible, and indicative of prognosis, for use as endpoints in clinical trials. PMID- 16217180 TI - Update on the Burkholderia cepacia complex. AB - PURPOSE OF REVIEW: The Burkholderia cepacia complex is comprised of a group of related bacterial species that are capable of causing life-threatening respiratory tract infection in persons with cystic fibrosis. This article reviews advances in our understanding of Burkholderia cepacia complex infection in cystic fibrosis, focusing on the taxonomy, clinical microbiology, and epidemiology, as well as the natural history and clinical outcomes associated with Burkholderia cepacia complex infection. RECENT FINDINGS: Each of the nine species of the Burkholderia cepacia complex has now received a formal species name. These names are the preferred nomenclature, replacing the former 'genomovar' designations. Studies from several countries reiterate that two species, Burkholderia cenocepacia and Burkholderia multivorans, account for most Burkholderia cepacia complex infection in cystic fibrosis. Bacterial genotyping studies indicate that specific Burkholderia cepacia complex strains infect multiple cystic fibrosis patients, implying that they may have an enhanced capacity for interpatient spread. Emerging clinical outcomes data suggest that at least some of these so called transmissible or epidemic strains are also more virulent in the cystic fibrosis host. Ongoing research is aimed at gaining a better understanding of Burkholderia cepacia complex ecology, defining Burkholderia cepacia complex virulence factors and pathogenic mechanisms, and determining the relative virulence of distinct strains. SUMMARY: Significant advances in our understanding of the Burkholderia cepacia complex serve as a critical foundation for further efforts that ultimately will enable better infection control and the development of novel therapeutics to treat Burkholderia cepacia complex infection in persons with cystic fibrosis. PMID- 16217182 TI - Behavioral aspects of nutrition in children with cystic fibrosis. AB - PURPOSE OF REVIEW: Nutritional status in cystic fibrosis is important to the health and long-term survival of patients; however, children with cystic fibrosis are not achieving the dietary recommendations or achieving normal growth. This paper reviews current knowledge about behavioral and environmental barriers to dietary adherence in this population. RECENT FINDINGS: Children with cystic fibrosis do not have the same appreciation of the need to gain weight as their parents. Parents of children with cystic fibrosis describe meal times as 'battling' with the child. Direct observation studies of meal times demonstrate that school-aged children with cystic fibrosis do not differ from children without cystic fibrosis on the frequency of behaviors incompatible with eating, as was seen in younger age groups, but parents of school-aged children with cystic fibrosis continue to engage in ineffective management styles at a greater frequency than parents of children without cystic fibrosis. Assessment of family functioning during mealtime demonstrates that these interaction patterns have a negative effect on family functioning in families of children with cystic fibrosis compared with families of same-age peers in all age groups: infancy/toddler, preschool, and school age. SUMMARY: Achieving the cystic fibrosis dietary recommendations is challenging, and typical parenting strategies to encourage eating not only are ineffective but may have a negative impact on family functioning. To improve dietary adherence and decrease conflict at meals, children with cystic fibrosis and their parents need to be taught more effective management strategies as part of dietary counseling. PMID- 16217183 TI - Bibliography. Current world literature. Sleep and respiratory neurobiology. PMID- 16217186 TI - Lauriston S. Taylor Lecture: Radiation protection in the aftermath of a terrorist attack involving exposure to ionizing radiation. AB - I would like to start this Twenty-Eighth Lauriston S. Taylor Lecture by expressing my gratitude to the National Council on Radiation Protection and Measurements (NCRP) for this unique occasion. I feel particularly honored for this opportunity to address a highly specialized and qualified audience of professionals who are specifically interested in what appears to be a forthcoming worldwide challenge, namely radiological terrorism and managing its potential radiological consequences. PMID- 16217188 TI - Warren K. Sinclair Keynote Address: Current challenges in countering radiological terrorism. AB - Terrorism, although perhaps known by other names, is not a new phenomenon. It dates back to Roman times and perhaps even further in world history. Caleb Carr says terrorism "is simply the contemporary name given to, and the modern permutation of, warfare deliberately waged against civilians with the purpose of destroying their will to support either their leaders or policies... " In modern times, in the United States, there have been isolated violent acts of citizens against each other, although these acts often were directed toward symbols of the federal government. In the Middle East and other parts of the world, acts of violence against U.S. citizens and military personnel date back into the early 1960's. Some of these acts seem to be almost random in nature. But these events occurred in distant lands of sometimes uncertain locations to the American public, who soon forgot them and their important message. Even though there had been at least one other attempt on the World Trade Center, it was not until 11 September 2001 that successful, large-scale acts of terrorism came to our shores. In 1998, the National Council on Radiation Protection and Measurements (NCRP) formed a Scientific Committee and charged the committee with the task of providing a report on the state of preparation and the potential use by terrorists of radiation and radioactivity. The draft report of the Committee was produced a year in advance of the events of 11 September 2001 and was published in its final form about a month after these terrible events. The report brought together, in one place, information that existed in a number of areas, not all of which were easily accessible. However, there were a number of gaps in the information and in the planning and preparation for such events. These were reflected in a series of recommendations for organization, planning, and training, as well as for research and development in a number of areas. This brief presentation will address a few selected areas that remain a challenge for those preparing for terrorist events involving radioactive materials. More detailed discussions will be provided by the presentations at this NCRP Annual Meeting. PMID- 16217189 TI - Radiological threat assessment and the Federal Response Plan--a gap analysis. AB - The ability of the federal government to effectively and efficiently respond to nuclear or radiological terrorist attacks has been the subject of intense discussion and analysis for many years. Because of recent terrorist activities and intelligence information, there is strong sentiment that it is not a question of if, but when, a radiological or nuclear terrorist attack will occur. As a result, there is considerable concern that the federal government may not be adequately prepared to respond to an attack involving a radiological dispersal device or improvised nuclear device. In response to these concerns, federal departments and agencies have initiated actions to develop a better understanding of the magnitude of the radiological/nuclear terrorist threat, assess the ability of the federal government to support state and local responses to such attacks, and improve the Nation's ability to prepare for, respond to, and recover from these types of attacks. In an era of limited fiscal growth and competing priorities, the federal government will have to enhance its collaboration with state and local governments, the private sector, and academia to ensure that the Nation is capable of responding to a terrorist attack involving radioactive or nuclear material. PMID- 16217191 TI - Advances in radiation detection technologies for responders. AB - The Department of Homeland Security is supporting the development of a large number of standards for first responders. In the area of detection of radioactive and nuclear materials, four new standards (ANSI N42.32, N42.33, N42.34, and N42.35) and their corresponding test and evaluation protocols were developed to meet Department of Homeland Security needs. Testing of the standards and protocols was carried out at the National Institute of Standards and Technology, Oak Ridge National Laboratory, Pacific Northwest National Laboratory, Los Alamos National Laboratory, and Lawrence Livermore National Laboratory. PMID- 16217190 TI - U.S. national response assets for radiological incidents. AB - The federal government has had the ability to respond to incidents of national significance for decades. Since 11 September 2001, there have been enhancements to existing federal assets and the creation of new federal assets. This presentation will provide an overview of the more significant federal assets. Pivotal to a response of national significance is the U.S. Department of Energy (DOE) Federal Radiological Monitoring and Assessment Center, which organizes and coordinates federal agency monitoring activities during an emergency. DOE manages the Federal Radiological Monitoring and Assessment Center during the emergency phase, and the Environmental Protection Agency (EPA) manages the response during the recovery phase once the emergency is terminated. EPA monitoring teams provide support during both the emergency and recovery phases of an emergency. Other DOE teams are available to respond to major nuclear power plant events, transportation accidents, or terrorism events involving the use of radiological materials, including the Radiological Assistance Program, the Aerial Measuring System, the National Atmospheric Release Advisory Center, and the Radiation Emergency Assistance Center/Training Site. For incidents involving a nuclear weapon, an improvised nuclear device, or a radiological dispersal device, DOE assets such as the Nuclear Emergency Support Team and the Accident Response Group could provide capabilities for weapon or device search, recovery, and removal. The Radiological Triage System harnesses the weapons scientists and engineers at the DOE national laboratories to provide gamma spectroscopy interpretation for agencies responding to an incident. In recent years, National Guard Weapons of Mass Destruction-Civil Support Teams have been created to support state and local response to terrorism events. The Civil Support Teams normally come under direct control of the state and can respond without requiring authorization from the U.S. Department of Defense (DoD). Changes brought about by the events of September 11 also extend to changing federal response policy and planning. Therefore, the Catastrophic Incident Response Annex to the National Response Plan is discussed. DoD also provides specifically designated radiological response capabilities that can be utilized within the guidelines of the National Response Plan. While optimally designed to support military missions, these resources also help provide for a well-equipped set of national assets to temporarily support and augment the local, state, and federal civil agencies that have primary authority and responsibility for domestic disaster assistance. The military's role in domestic emergencies is well defined in military regulations, as well as the national plan. PMID- 16217192 TI - Medical resources and requirements for responding to radiological terrorism. AB - Medical planning and response to radiological terrorism is different than planning or responding to an event such as a nuclear power plant accident. The major differences are that now we must plan for multiple simultaneous events, suicide scenarios, and the possibility of biological, chemical, and radiological agents being used at the same time. This demands an "all-hazards" approach and not just a radiological response. An overview of the issues related to diagnosis, treatment, training, and resources is provided. Although the requirements for medical management are clear, the available resources have not been applied in a manner that results in adequate preparedness for radiological events. PMID- 16217194 TI - Medical management of radiological casualties. AB - Victims of radiological terrorism events require prompt diagnosis and treatment of medical and surgical conditions as well as conditions related to radiation exposure. Hospital emergency personnel should triage victims using traditional medical and trauma criteria. Radiation dose can be estimated early post-event using rapid-sort, automated biodosimetry and clinical parameters such as the clinical history, the time to emesis (TE), and lymphocyte depletion kinetics. For TE < 2 h, the effective whole-body dose is at least 3 Gy. If TE < 1 h, the whole body dose most probably exceeds 4 Gy. Lymphocyte depletion follows dose dependent, first order kinetics after high-level gamma and criticality incidents. Patient radiation dose can be estimated very effectively from the medical history, serial lymphocyte counts, and TE, and subsequently confirmed with chromosome-aberration bioassay, the current gold standard. These data are effectively analyzed using the Armed Forces Radiobiology Research Institute Biodosimetry Assessment Tool. The medical management of patients with acute, moderate to severe radiation exposure (effective whole-body dose >3 Gy) should emphasize the rapid administration of colony stimulating factors. All of these compounds decrease the duration of radiation-induced neutropenia and stimulate neutrophil recovery, albeit with some variability, in patients who have received myelotoxic chemotherapy, and all have demonstrated benefit in irradiated animals. For those patients developing febrile radiation-induced neutropenia, adherence to the current Infectious Diseases Society of America guidelines for high-risk neutropenia is recommended. PMID- 16217193 TI - Early-response biological dosimetry--recommended countermeasure enhancements for mass-casualty radiological incidents and terrorism. AB - The effective medical management of a suspected acute radiation overexposure incident necessitates recording dynamic medical data, measuring appropriate radiation bioassays, and estimating dose from dosimeters and radioactivity assessments in order to provide diagnostic information to the treating physician and a dose assessment for personnel radiation protection records. The accepted generic multiparameter and early-response approach includes measuring radioactivity and monitoring the exposed individual; observing and recording prodromal signs/symptoms and erythema; obtaining complete blood counts with white blood cell differential; sampling blood for the chromosome-aberration cytogenetic bioassay using the "gold standard" dicentric assay (translocation assay for long times after exposure) for dose assessment; bioassay sampling, if appropriate, to determine radioactivity contamination; and using other available dosimetry approaches. In the event of a radiological mass-casualty incident, current national resources need to be enhanced to provide suitable dose assessment and medical triage and diagnoses. This capability should be broadly based and include stockpiling reagents and devices; establishing deployable (i.e., hematology and biodosimetry) laboratories and reference (i.e., cytogenetic biodosimetry, radiation bioassay) laboratories; networking qualified reference radioactivity counting bioassay laboratories, cytogenetic biodosimetry, and deployable hematology laboratories with the medical responder community and national radiation protection program; and researching efforts to identify novel radiation biomarkers and develop applied biological dosimetry assays monitored with clinical, deployable, and hand-held analytical systems. These research and applied science efforts should ultimately contribute towards approved, regulated biodosimetry devices or diagnostic tests integrated into a national radioprotection program. PMID- 16217195 TI - Hospital management of mass radiological casualties: reassessing exposures from contaminated victims of an exploded radiological dispersal device. AB - One of the key issues in the aftermath of an exploded radiological dispersal device from a terrorist event is that of the contaminated victim and the concern among healthcare providers for the harmful exposures they may receive in treating patients, especially if the patient has not been thoroughly decontaminated. This is critically important in the event of mass casualties from a nuclear or radiological incident because of the essential rapidity of acute medical decisions and that those who have life- or limb-threatening injuries may have treatment unduly delayed by a decontamination process that may be unnecessary for protecting the health and safety of the patient or the healthcare provider. To estimate potential contamination of those exposed in a radiological dispersal device event, results were used from explosive aerosolization tests of surrogate radionuclides detonated with high explosives at the Sandia National Laboratories. Computer modeling was also used to assess radiation dose rates to surgical personnel treating patients with blast injuries who are contaminated with any of a variety of common radionuclides. It is demonstrated that exceptional but plausible cases may require special precautions by the healthcare provider, even while managing life-threatening injuries of a contaminated victim from a radiological dispersal device event. PMID- 16217196 TI - Addressing the psychosocial and communication challenges posed by radiological/nuclear terrorism: key developments since NCRP Report No. 138. AB - One of the most innovative aspects of NCRP Report No. 138 (Management of Terrorist Incidents Involving Radioactive Material) was the high priority it accorded to psychosocial and communication issues. While previous discussions of radiological and nuclear terrorism had occasionally referred to these topics, NCRP Report No. 138 was the first report of its kind to recognize the profound challenges posed by these issues and to place them at the heart of preparedness and response efforts. In the years since the report's release, a host of important developments have taken place in relation to psychosocial and communication issues. This paper reviews key changes and advances in five broad areas: (1) training exercises, (2) policy and guidance development, (3) findings on hospital and clinician needs, (4) survey research on public perceptions of radiological terrorism, and (5) risk communication for radiological and nuclear terrorism situations. The article concludes with a discussion of continuing psychosocial and communication challenges, including critical areas needing further attention as the nation moves to meet the threat of terrorism involving radioactive materials. PMID- 16217197 TI - Radiation protectants: current status and future prospects. AB - In today's heightened nuclear/biological/chemical threat environment, there is an increased need to have safe and effective means to protect not only special high risk service groups, but also the general population at large, from the health hazards of unintended ionizing radiation exposures. An unfulfilled dream has been to have a globally effective pharmacologic that could be easily taken orally without any undue side effects prior to a suspected or impending nuclear/radiological event; such an ideal radioprotective agent has yet to be identified, let alone fully developed and approved for human use. No one would argue against the fact that this is problematic and needs to be corrected, but where might the ultimate solution to this difficult problem be found? Without question, representative species of the aminothiol family [e.g., Amifostine (MedImmune, Gaithersburg, Maryland)] have proven to be potent cytoprotectants for normal tissues subjected to irradiation or to radiomimetic chemicals. Although Amifostine is currently used clinically, drug toxicity, limited times of protection, and unfavorable routes of administration, all serve to limit the drug's utility in nonclinical settings. A full range of research and development strategies is being employed currently in the hunt for new safe and effective radioprotectants. These include: (1) large scale screening of new chemical classes or natural products; (2) restructuring/reformulating older protectants with proven efficacies but unwanted toxicities; (3) using nutraceuticals that are only moderately protective but are essentially nontoxic; (4) using low dose combinations of potentially toxic but efficacious agents that protect through different routes to foster radioprotective synergy; and (5) accepting a lower level of drug efficacy in lieu of reduced toxicity, banking on the premise that the protection afforded can be leveraged by post-exposure therapies. Although it is difficult to predict which of these strategies will ultimately prove to be successful, it is certain that the probability of a useful protectant being fielded is increased significantly. This is due to the resurgence of interest in radiation protection, increased resources being expended by federal agencies, and by the Food and Drug Administration's willingness to innovate relative to new approval guidance. PMID- 16217198 TI - Defining the full therapeutic potential of recombinant growth factors in the post radiation-accident environment: the effect of supportive care plus administration of G-CSF. AB - Radiation accidents are uncontrolled and ill-defined, the exposure is nonuniform and may be partial body, forecasting a variable dose distribution and sparing of hematopoietic stem cells. We propose that the "best" treatment protocol available now for severely irradiated personnel with acute hematopoietic syndrome is the combination of supportive care and administration of recombinant cytokines as soon as possible after irradiation. Herein, we demonstrate the significant effect of G-CSF administration on lethally irradiated canines. G-CSF administered early and continuously after irradiation increased neutrophil recovery and survival over a lethal and supralethal dose range. The respective LD50/30 for the control, supportive care-alone cohorts of 338 cGy, was increased to 488 cGy with administration of G-CSF. Clearly, the use of supportive care and G-CSF enhanced recovery of myelopoiesis and survival after lethal doses of irradiation. PMID- 16217199 TI - International efforts in countering radiological terrorism. AB - An analysis of the possible goals of terrorists and the means of achieving them leads to an assessment of the most likely target materials and facilities for radiological terrorism. International efforts in countering these terrorist goals cover several objectives. The first is to prevent acquisition of the target radioactive materials, or access to the target facilities. This has to be achieved throughout the life cycle for radioactive sources. The second objective is to detect attempts to acquire, or actual acquisition of, radioactive materials. The third objective is to effectively detect and respond to the use (or threat of use) of these materials, or sabotage of the facilities. An additional goal is to minimize the consequences of any such use or sabotage. The number of international bodies, groups, agencies, and organizations contributing to these goals is very large and almost impossible to catalogue. However, this paper groups the types of efforts that are being undertaken, identifies the key agencies involved, and provides some specific examples of their work. PMID- 16217200 TI - Chernobyl and Goiania lessons for responding to radiological terrorism. AB - The deployment of a radiological dispersal device (RDD) is likely to result in relatively low radiation exposure of the targeted population, insufficient to cause a severe radiation detriment. Nevertheless, due to atmospheric dispersion of the radioactive material, an urban area equaling several city blocks could be affected. The current knowledge base concerning the response to radiological terrorism, focusing mainly on environmental cleanup and site recovery (CSR) of areas with radioactive contamination due to the deployment of an RDD, is largely derived from military scientific tests or exercises assembled over the past 50 y with only limited applicability to the consequences of an RDD detonating in a city. This paper focuses on the extensive experience in CSR gained in the management of the radiological accident contaminating the Brazilian city of Goiania in 1987, and managing the aftermath of the Chernobyl reactor accident in 1986. The incident in Goiania demonstrated the numerous practical difficulties of implementing a sound CSR, based on a balanced judgment of all relevant factors, such as radiation safety, environmental issues, economic consequences, and public fear. A review of the different stages of the intervention policy in the former Soviet Union reveals that risk-benefit cost analysis was not used for the decision-making process during the later stages of the post-accident situation. Instead, a CSR policy was adopted that resulted in continuously escalating costs. The results of this analysis are used to develop an Integrated Cleanup and Site Restoration Concept and recommend practically applicable solutions from Lessons Learned. PMID- 16217201 TI - Proposed framework for cleanup and site restoration following a terrorist incident involving radioactive material. AB - Cleanup following a terrorism incident involving a radiological dispersal device (RDD) or improvised nuclear device (IND) is likely to be technically challenging, costly, and politically charged. Lessons learned from the Top Officials 2 exercise and the increased threat of terrorist use of an RDD or IND have driven federal officials to push for an agreed-upon process for determining appropriate cleanup levels. State and local authorities generally have the ultimate responsibility for final public health decisions in their jurisdictions. In response to terrorist attacks, local authorities are likely to request federal assistance in assessing the risk and establishing appropriate cleanup levels. It is realistic to expect local and state requests for significant federal assistance in planning and implementing recovery operations. State and local authorities may desire "shared accountability" with the federal government in setting the appropriate cleanup levels. Government officials at all levels will face pressure to say how clean is clean enough and how quickly people can re enter affected areas. Issues arising include (1) the nature of the relationship between the federal, state, and local leadership involved in the recovery efforts and (2) where the funding for recovery comes from. Many agencies, including the U.S. Environmental Protection Agency (EPA), the U.S. Nuclear Regulatory Commission (NRC), and the U.S. Department of Energy (DOE) have long been involved in cleanup activities involving radioactive materials. These agencies have recognized the need for a participatory process and realize the need to remain flexible when faced with possible unprecedented environmental challenges following a terrorist attack. Currently, the Department of Homeland Security has a committee process underway, with participation of the EPA, NRC, DOE, and other federal agencies, to try to resolve these issues and to begin engaging state, local, and tribal governments, and others as appropriate. PMID- 16217202 TI - Practical and scientifically based approaches for cleanup and site restoration. AB - This paper presents practical and scientific approaches for cleanup and site restoration following terrorist events. Both approaches are required in actual emergency situations and are complementary. The practical examples are taken from the May 2003 second biannual national emergency exercise, Top Officials 2 (TOPOFF 2), which occurred in Chicago, Illinois, and Seattle, Washington. The scientific examples are taken from the Department of Energy sites at Rocky Flats, Fernald, and Los Alamos where cleanup initiatives based on scientific approaches and community input are underway. Three examples are provided to explain, from a practical standpoint, how decisions during the exercise had to be made quickly, even though the alternatives were not always clear. These examples illustrate how scientific approaches can be integrated into the resolution of these dilemmas. The examples are (1) use of water to wash city roads and freeways contaminated with plutonium, Am, and Cs; (2) decontamination of large public ferries that passed through a radioactive plume; and (3) handling of wastewater following decontamination within a city. Each of these situations posed the need for an immediate decision by authorities in charge, without the benefit of community input or time for an analysis of the important pathways of exposure. It is evident there is a need to merge the practical knowledge gained in emergency response with scientific knowledge learned from cleanup and site restoration. The development of some basic scientific approaches ahead of time in the form of easy to-use tools will allow practical decisions to be made more quickly and effectively should an actual terrorist event occur. PMID- 16217209 TI - Taking your patient's spiritual vital signs. PMID- 16217210 TI - A case study in home health disease management. PMID- 16217211 TI - Efficacy of influenza vaccine in the elderly. PMID- 16217213 TI - Interviews with patients who have cancer and their family members provide insight for clinicians. AB - The words of patients with cancer about pain management are powerful learning tools. Questions were posed about the meaning of pain for patients and families, effective pain management strategies, and professional caregivers' roles to three patients and families. This article details their responses about pain, pain management, and the barriers they encountered. Clinicians are urged to read these case reports to "hear" the importance of effective pain management to individuals and act appropriately. PMID- 16217214 TI - Home care nurses' perceptions of control over cancer pain. AB - This qualitative study examined home care nurses' perceptions of control over cancer pain. Four major themes emerged: Being heard, feeling invisible in the pain management process; not knowing, a need for pain education; control through advocacy; and patient-related barriers to optimal pain management. This study documents the need for continued education in pain management and communication skills for home care nurses. PMID- 16217216 TI - Episodic case management in home care. AB - Home care clinicians are being given increasing responsibility for managing a patient's care from admission to discharge and beyond. Under PPS, case management oversight using model(s) unique to an agency's case mix is critical to the financial and quality outcomes of the entire organization. This article provides the latest thinking on case management and how agencies and clinicians can manage cases successfully. PMID- 16217217 TI - Opportunities in care coordination. AB - The growing incidence of chronic illness and the increase in the number of elderly individuals wishing to live in their own homes as long as possible have created opportunities for care coordination services. Home care providers are well positioned to take advantage of the need for care coordination. The developing nature of the market for these services means providers ready to offer care coordination can influence how care coordination services will be packaged, delivered, and funded. PMID- 16217219 TI - A tribute to hospice. PMID- 16217220 TI - M0488: status of surgical wound. PMID- 16217224 TI - AACVPR: the first 20 years. PMID- 16217225 TI - Quality indicators in cardiovascular care: the case for cardiac rehabilitation. PMID- 16217226 TI - Independent effect of cardiac rehabilitation on lipids in coronary artery disease. AB - PURPOSE: We evaluate the effects cardiac rehabilitation (CR) participation independent of using lipid-altering agents (LAAs) on low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, non-high-density lipoprotein cholesterol, total cholesterol, and triglyceride. Measurements included absolute and relative change in lipids and increases in percent of patients achieving goals. METHODS: Analysis of 766 patients who participated in CR between 2000 and 2003 was performed. On enrollment to CR, all were being treated with an LAA defined as HIviG-CoA reductase inhibitors, bile acid sequestrant, fibrate, and niacin, hormone replacement therapy. Preenrollment and postenrollment lipids were obtained. Analysis was performed on 2 cohorts, participants enrolled on an LAA with no change in medication (n = 13) and participants enrolled on an LAA with a change in medications (n = 153). RESULTS: At completion of CR, 74.9% of patients on LAA at enrollment with no medication adjustments during the program were at Adult Treatment Panel III goal for low-density lipoprotein cholesterol compared with 68.5% at entry (P = .01), all other lipid parameters also significantly improved. Sixty-three percent who started CR on an LAA and had dose adjustment or an additional LAA added achieved low-density lipoprotein cholesterol goal compared with 43.1% at entry (P < .0001). CONCLUSION: Participation in CR significantly potentiates the lipid-improving effects of pharmacological therapy and independently contributed to the percent of patients achieving all lipid levels at Adult Treatment Panel III goal. PMID- 16217228 TI - Cardiac rehabilitation and getting to lipid goals. PMID- 16217229 TI - Validity of hand-held dynamometry for strength assessment in cardiac rehabilitation. PMID- 16217230 TI - Effects of horticultural therapy on mood and heart rate in patients participating in an inpatient cardiopulmonary rehabilitation program. AB - PURPOSE: To assess the effects of horticultural therapy (HT) on mood state and heart rate (HR) in patients participating in an inpatient cardiac rehabilitation program. METHODS: Cardiac rehabilitation inpatients (n = 107) participated in the study. The HT group consisted of 59 subjects (34 males, 25 females). The control group, which participated in patient education classes (PECs), consisted of 48 subjects (31 males, 17 females). Both HT sessions and PEC are components of the inpatient rehabilitation program. Each group was evaluated before and after a class in their respective modality. Evaluation consisted of the completion of a Profile of Mood States (POMS) inventory, and an HR obtained by pulse oximetry. RESULTS: Changes in the POMS total mood disturbance (TMD) score and HR between preintervention and postintervention were compared between groups. There was no presession difference in either TMD score (16 +/- 3.6 and 19.0 +/- 3.2, PEC and HT, respectively) or HR (73.5 +/- 2.5 and 79 +/- 1.8, PEC and HT, respectively). Immediately following the intervention, the HT TMD was significantly reduced (post-TMD = 1.6 +/- 3.2, P < .001), while PEC TMD was not significantly changed (TMD = 17.0 +/- 28.5). After intervention, HR fell in HT by 4 +/- 9.6 bpm (P < .001) but was unchanged in PEC. CONCLUSION: These findings indicate that HT improves mood state, suggesting that it may be a useful tool in reducing stress. Therefore, to the extent that stress contributes to coronary heart disease, these findings support the role of HT as an effective component of cardiac rehabilitation. PMID- 16217231 TI - Impact of brief or extended exercise training on the benefit of a dyspnea self management program in COPD. AB - PURPOSE: To evaluate the differences in the long-term outcomes of dyspnea, exercise performance, health-related quality of life, and health resource utilization following a dyspnea self-management program with 3 different "doses" of supervised exercise. PATIENTS AND METHODS: In a prospective, randomized, single-blind, 1-year trial, patients with stable chronic obstructive pulmonary disease (N = 103; age 66 +/- 8, females 57; FEV1 44.8% +/- 14% predicted) were randomly assigned to either: (1) Dyspnea self-management program (DM); (2) DM plus 4 supervised exercise sessions (DM-exposure); or (3) DM plus 24 supervised exercise sessions (DM-training). The dyspnea self-management program included individualized education and demonstration of dyspnea self-management strategies, an individualized home walking prescription, and biweekly nurse telephone calls. Outcomes were measured at baseline and every 2 months for 1 year. RESULTS: The DM training group had significantly greater improvements in dyspnea during incremental treadmill test and in exercise performance on the incremental and endurance treadmill tests at 6 and 12 months compared with the other 2 groups. Dyspnea with activities of daily living and self-reported physical functioning significantly improved for all groups over time. The dose-response relationship between supervised exercise and improvement in dyspnea present at 2 months was not sustained over the year. CONCLUSION: Consistent with previous findings from evaluation studies of pulmonary rehabilitation programs, the greater number of supervised exercise training sessions improved laboratory dyspnea and performance more than the other two doses of exercise. In the long term, the improvement in dyspnea with activities of daily living and physical functioning was similar for all 3 groups. PMID- 16217232 TI - More is better once again! PMID- 16217236 TI - Stress testing in the medical evaluation of Hazmat workers. PMID- 16217237 TI - Hexavalent chromium study's conclusions unjustified. PMID- 16217238 TI - On an ethic of occupational and environmental medicine: response to doctors Guidotti, Lippin, Key, and anonymous. PMID- 16217241 TI - Mortality among semiconductor and storage device-manufacturing workers. AB - PROBLEM: We evaluated mortality during 1965 to 1999 among 126,836 workers at two semiconductor facilities and one storage device facility. METHOD: We compared employees' cause-specific mortality rates with general population rates and examined mortality patterns by facility, duration of employment, time since first employment, and work activity. RESULTS: Employees had lower-than-expected mortality overall (6579 observed deaths, standardized mortality ratio [SMR] = 65; 95% confidence interval [CI] = 64-67), for all cancers combined (2159 observed, SMR = 78, 95% CI = 75-81) and for other major diseases. Central nervous system cancer was associated with process equipment maintenance at one of the semiconductor facilities (10 observed, SMR = 247, 95% CI = 118-454). Prostate cancer was associated with facilities/laboratories at the storage device facility (18 observed, SMR = 198, (5% CI = 117-313). CONCLUSIONS: Further evaluation of workplace exposures or independent investigations of similar occupational groups may clarify the interpretation of associations observed in this study. PMID- 16217240 TI - Exposure assessment for retrospective follow-up studies of semiconductor- and storage device-manufacturing workers. AB - OBJECTIVE: This exposure assessment was conducted in the first large study of mortality and cancer incidence in semiconductor and storage device manufacturing. METHODS: Unique combinations of division, department and job codes and names (DDJ) from work history records were assigned to work groups and exposure categories. Agent exposure matrices assessed differences in potential exposures between groups. Changes in exposure over time were tracked by dividing the production history into manufacturing eras. RESULTS: Nineteen work groups were developed to capture 310,351 unique DDJs from 1965-1999. Agent exposure matrices contrasted exposure potential to solvents, metals, and work in cleanrooms between groups, and three manufacturing eras were identified for each site. CONCLUSIONS: The work groups, manufacturing eras and agent matrices have been used to classify workers in the study of cancer incidence and mortality. PMID- 16217242 TI - Psychosocial work characteristics as predictors for burnout: findings from 3-year follow up of the PUMA Study. AB - OBJECTIVE: The objective of this study was to investigate the impact of psychosocial work characteristics on burnout. METHODS: A total of 1772 participants in different human service sector organizations were eligible for the cross-sectional analyses (baseline) and 952 for the prospective analyses. We measured 14 psychosocial work characteristics and three types of burnout. Linear regression models were used for analyzing associations between psychosocial work characteristics at baseline and burnout at baseline and at 3 years of follow up. RESULTS: Low possibilities for development, high meaning of work, low predictability, high quality of leadership, low role clarity, and high role conflicts predicted burnout at 3 years of follow up after the psychosocial work characteristics were adjusted for each other, potential confounders, and burnout level at baseline. CONCLUSION: Psychosocial work characteristics were prospectively associated with burnout, suggesting that improving the psychosocial work environment may reduce future burnout in human service work. PMID- 16217243 TI - A randomized, double-blind placebo-controlled trial evaluating the effects of vitamin E and selenium on arsenic-induced skin lesions in Bangladesh. AB - OBJECTIVE: We sought to determine whether supplementation of vitamin E (alpha tocopherol), selenium (L-selenomethionine), or their combination improves arsenical skin lesions. METHODS: A 2 x 2 randomized, placebo-controlled, double blind trial among 121 men and women chronically exposed to arsenic in drinking water was conducted in rural Bangladesh. Participants were randomized to one of four treatment arms: vitamin E, selenium, vitamin E and selenium (combination), or placebo and were treated for 6 months. RESULTS: At baseline, the average skin lesion scores were 2.23, 2.26, and 2.63 and at follow-up, the average skin lesion scores went down to 2.00, 2.06, and 2.47 in those receiving vitamin E, selenium, and the combination, respectively. CONCLUSIONS: Supplementation with vitamin E and selenium, either alone or in combination, slightly improved skin lesion status, although the improvement was not statistically significant. PMID- 16217244 TI - Perceptions of provider communication and patient satisfaction for treatment of acute low back pain. AB - OBJECTIVE: We sought to assess the relationship between perceptions of provider communication and treatment satisfaction for acute, work-related low-back pain (LBP). METHODS: In a prospective cohort study, 544 working adults (67% men) with acute LBP provided 1- and 3-month assessments of pain, function, and work status. RESULTS: In a multiple regression analysis, positive provider communication (took problem seriously, explained condition clearly, tried to understand my job, advised to prevent re-injury) explained more variation in patient satisfaction at 1 month than was explained by clinical improvements in pain and function. At 3 months, clinical improvement variables surpassed provider communication as predictors of patient satisfaction. CONCLUSIONS: Patients with work-related LBP place a high value on provider counseling and education, especially during the acute stage (<1 month) of treatment. PMID- 16217245 TI - Incidence of carpal tunnel syndrome among automobile assembly workers and assessment of risk factors. AB - OBJECTIVES: This study defined the incidence rate of carpal tunnel syndrome (CTS) among active assembly workers and evaluated risk factors. METHODS: This study followed 189 automobile assembly workers over 1 year. Incident cases were defined as workers who had no current or prior history of CTS and were subsequently diagnosed with CTS by means of active surveillance during the study. RESULTS: The estimated annual incidence rate ranged from 1% to 10%, depending on the case definition. Significant predictors for CTS include a higher baseline median-ulnar peak latency difference, history of wrist/hand/finger tendonitis, history of diabetes, nonneutral wrist and elbow postures, and a lower self-reported social support. CONCLUSIONS: This is similar to other prospective studies that demonstrate both ergonomic and medical history are independent risk factors for development of CTS. PMID- 16217246 TI - A review and analysis of the clinical and cost-effectiveness studies of comprehensive health promotion and disease management programs at the worksite: update VI 2000-2004. AB - This critical review focuses on the 12 new studies focused on the clinical and cost outcomes research focused on worksites and published between 2000 and 2004. Although these new studies indicate further evidence of positive clinical and cost outcomes, the quantity and quality of such research continue to decline. When corporations and health plans are demanding more evidence-based outcomes, this decline in rigorous research marks a serious challenge to the field of health promotion and disease management. PMID- 16217247 TI - Evaluation of epidemiologic and animal data associating pesticides with Parkinson's disease. AB - Exposure to pesticides may be a risk factor for developing Parkinson's disease (PD). To evaluate the evidence regarding this association in the scientific literature, we examined both analytic epidemiologic studies of PD cases in which exposure to pesticides was queried directly and whole-animal studies for PD-like effects after systemic pesticide exposure. Epidemiologic studies were considered according to study quality parameters, and results were found to be mixed and without consistent exposure-response or pesticide-specific patterns. These epidemiologic studies were limited by a lack of detailed and validated pesticide exposure assessment. In animal studies, no pesticide has yet demonstrated the selective set of clinical and pathologic signs that characterize human PD, particularly at levels relevant to human populations. We conclude that the animal and epidemiologic data reviewed do not provide sufficient evidence to support a causal association between pesticide exposure and PD. PMID- 16217250 TI - Once in a lifetime. PMID- 16217251 TI - Cure is not enough. PMID- 16217252 TI - Nasopharyngeal carcinoma in the pediatric age group: the northern Israel (Rambam) medical center experience, 1989-2004. AB - Nasopharyngeal carcinoma (NPC) is rare in children, accounting for less than 1% of all malignancies. Radiation therapy has been the mainstay of treatment of many years, but to improve survival, the use of chemotherapy has been advocated. This is a retrospective analysis of 13 patients less than 20 years of age treated for NPC the Rambam Medical Center during 1989 to 2004. Eight boys and five girls with a median age of 14.5 years (range 10-19) were included. Median follow up (including patients who died) was 6.15 years (range 1-15 years). Duration of symptoms was 1 to 24 months (median 5 months). Of the 13 patients, one patient had stage I, 6 had stage III, 5 had stage IV-A, and 1 had stage IV-B disease. Ten patients (77%) had undifferentiated carcinoma (WHO type III) and three patients (23%) had nonkeratinizing carcinoma (WHO type II). Most of the children received two or three courses of neoadjuvant multiagent chemotherapy consisting of cisplatin and 5-FU, followed by radiotherapy with doses in excess of 60 Gy. One child received concomitant chemoradiation. Ten of the 13 patients (77%) are alive without disease 6 years after diagnosis (range 1-15 years). One patient developed local and distant metastases 1 year after diagnosis and is currently receiving combined radiochemotherapy. Two patients died. Overall survival was 84%; event free survival was 77%. Nine patients (69%) developed moderate to severe long-term complications. Pediatric NPC is curable by combined radiation and chemotherapy, with doses of radiation in excess of 60 Gy. Long-term follow-up is important for early detection of second malignancies as well as for radiation-induced endocrinologic deficiencies and other normal tissue complications. PMID- 16217253 TI - Second malignancy in 597 patients with ewing sarcoma of bone treated at a single institution with adjuvant and neoadjuvant chemotherapy between 1972 and 1999. AB - The relative risk of second tumors in patients with Ewing sarcoma is controversial, and little is known about their treatment and outcome. The purpose of the current study was to define the incidence and features of second tumors among 597 long-term survivors of nonmetastatic Ewing sarcoma treated with adjuvant and neoadjuvant chemotherapy, radiotherapy, and/or surgery. The authors found that the risk of secondary malignancy after adjuvant or neoadjuvant treatment of Ewing sarcoma is higher than that after other childhood or adolescent cancers only after radiotherapy. Based on this, postoperative radiotherapy should be avoided when surgery with adequate margins is feasible. PMID- 16217254 TI - Wilms tumor in monozygous twins: clinical, pathological, cytogenetic and molecular case report. AB - The concomitant occurrence of Wilms tumor (WT) was observed in two monozygotic twin sisters without evidence of congenital malformations. Twin 1 was diagnosed with a stage I WT at 11 months of age, whereas twin 2 developed a bilateral (stage V) WT at 13 months of age. In both cases pathologic examination showed a nonanaplastic stromal type WT, with marked rhabdomyomatous elements. Cytogenetic analyses performed on blood samples and on tumor specimens revealed no karyotypic abnormality. No alteration of the WT1 and POU6F2 genes was identified in constitutional and tumor DNA of both sisters, and no anomaly in WT1 expression was evidenced in the normal kidney of one of them. However, loss of heterozygosity on chromosome 11p, involving the alleles of maternal origin, was detected both in the single tumor of twin 1 and in the two distinct tumors of twin 2, thus suggesting a common etiology of the diseases. To the authors' knowledge, this is the first report describing at both the clinical and genetic level a couple of monozygotic twins concordant for WT development. PMID- 16217255 TI - Risk score for pediatric intensive care unit admission in children undergoing hematopoietic stem cell transplantation and analysis of predictive factors for survival. AB - The authors retrospectively analyzed postransplantation events in 198 children who underwent hematopoietic stem cell transplantation (HSCT) between 1998 and 2002 to obtain a risk score for pediatric intensive care unit (PICU) admission and to ascertain variables predicting a poor outcome. Thirty-six patients (18%) were admitted to the PICU. Median age was 9 years (range 1-18). On univariate analysis, variables significantly associated with PICU admission were male gender (P = 0.01), more than first complete remission (P = 0.003), allogeneic transplantation (P = 0.001), engraftment syndrome (P = 0.03), and acute graft versus-host disease grade of at least two (P = 0.05). According to this, patients were divided in two levels of risk (low and high), with a respective probability of PICU admission of 8.8 +/- 2.2% and 63.8 +/- 8.8% (P < 0.0001). Seventeen (47%) patients were discharged from the PICU. The probability of event-free survival after PICU admission at 3 years was 24.2 +/- 7%. On univariate analysis, variables with a negative impact on event-free survival were type of transplantation, inotropic support, a C-reactive protein level of at least 10 mg/dL, and a high O-PRISM score. On multivariate analysis, the only variable that influenced event-free survival was the O-PRISM score (< or =10 points, 54.6 +/- 15.3%; >10 points, 8.6 +/- 5.8%; P = 0.007). In conclusion, the risk of PICU admission may be easily estimated using simple variables. A high O-PRISM score at the time of PICU admission predicts a dismal outcome. PMID- 16217256 TI - Antithymocyte globulin pharmacokinetics in pediatric patients after hematopoietic stem cell transplantation. AB - To analyze the dose effects of rabbit-derived antithymocyte globulin (ATG) in children after allogeneic hematopoietic stem cell transplantation (HSCT), ATG serum levels were monitored in 32 children and adolescents (median age 3.42 years, range 0.34-18.67 years) and the incidence of acute and chronic graft versus-host disease, rejection, viral infections, EBV-lymphoproliferative disease, and survival was correlated with the ATG dose used. Cumulative doses from 7.5 to 20 mg/kg showed a constant half-life and linear correlation between dose and Cmax, whereas higher doses (30-40 mg/kg) accumulated in the body. High dose ATG is of no benefit for preventing graft-versus-host disease but is associated with a significant increase in EBV-linked disease, and it appears to enhance the susceptibility to fatal viral infections and rejection. These data strongly support the use of a low-dose ATG regimen in pediatric HSCT. PMID- 16217257 TI - Gene expression profile of ewing sarcoma cell lines differing in their EWS-FLI1 fusion type. AB - The t(11;22)(q24;q12) translocation is present in up to 95% of Ewing tumor patients and results in the formation of an EWS-FLI-1 fusion gene that encodes a chimeric transcription factor. Many alternative forms of EWS-FLI-1 exist because of variations in the location of the EWS and FLI-1 genomic breakpoints. Previous reports have shown that the type 1 fusion is associated with a significantly better prognosis than the other fusion types. It has been suggested that the observed clinical discrepancies result from different transactivation potentials of the various EWS-FLI-1 fusion proteins. In an attempt to identify genes whose expression levels are differentially modulated by structurally different EWS-FLI 1 transcription factors, we have used microarray technology to interrogate 19,000 sequence genes to compare gene expression profile of type 1 or non-type 1 Ewing sarcoma cell lines. Data analysis showed few qualitative differences on gene expression; expression of only 41 genes (0.215% of possible sequences analyzed) differed significantly between Ewing tumor cell lines carrying EWS-FLI-1 fusion type 1 with respect to those with non-type 1 fusion. PMID- 16217258 TI - Will the next generation of "safer" cigarettes be safer? AB - There are three basic means of avoiding smoking-related diseases: never starting to smoke, quitting, and avoiding smoke-filled environments. Recently, a fourth possibility has emerged: use of new, ostensibly less toxic products by smokers who cannot or will not quit, including "reduced toxin" cigarettes and novel smokeless tobacco products. To their purveyors, these new "tobacco harm reduction" (THR) products represent an opportunity for inveterate smokers to reduce their risk of lung cancer and other diseases. To health professionals, the new products pose a myriad of risks. This new generation of THR products is not the first to promise reduced risk, however. Both filtered cigarettes and low tar and nicotine cigarettes were marketed with explicit health themes, ultimately with disastrous results for public health. THR products enter the market subject to no product regulation whatsoever; thus, the opportunity for objective, independent scientific evaluation of their risks and benefits, and for regulation of advertising or sale as a result, is absent. This paper describes the new generation of THR products, discusses potential benefits and risks, examines lessons from the experience with filtered and low tar and nicotine cigarettes, and describes the principal challenges that confront the medical profession, government, and the public in determining what to do with this perplexing array of novel products. PMID- 16217259 TI - Fulminant hepatitis due to varicella zoster virus in a girl with acute lymphoblastic leukemia in remission: report of a case and review. AB - The authors describe a 4-year-old girl with acute lymphoblastic leukemia in remission who developed fulminant hepatic failure due to varicella-zoster virus (VZV). Diagnosing VZV visceral infection in immunocompromised patients is often difficult due to atypical clinical presentation with few or no skin lesions and severe abdominal or back pain. Prompt initiation of empirical treatment with acyclovir and VZV immunoglobulin pending results of the serum polymerase chain reaction for VZV is warranted in this clinical setting. PMID- 16217260 TI - Acquired Glanzmann's thrombasthenia associated with acute lymphoblastic leukemia. AB - Acquired Glanzmann's thrombasthenia is an uncommon event in association with leukemia. The authors describe a patient with acute lymphoblastic leukemia (ALL) who presented with severe hemorrhagic syndrome, without disseminated intravascular coagulation. The patient's course was complicated by the occurrence of severe hemorrhagic episodes, with a thrombasthenia-like profile, requiring multiple transfusions with packed red cells, platelets, and fresh-frozen plasma. Biological explorations detected anti-GPIIb/IIIa complex antibodies. The patient finally died with refractory disease and persistent bleeding. This case is the first reported of autoantibodies to GPIIb/IIIa in ALL. Such paraneoplastic syndrome is potentially responsible for severe life-threatening hemorrhage. PMID- 16217261 TI - A spontaneous intramural hematoma of the bowel presenting as obstruction in a child receiving low-molecular-weight heparin. AB - Low-molecular-weight heparin (LMWH) is a safe and effective alternative to unfractionated heparin and coumadin in the treatment and prophylaxis of thrombosis in children. When compared with these more established anticoagulants, it is easier to achieve therapeutic levels and the incidence of hemorrhagic complications is equivalent or lower. In children there is less published experience than in adults, but the low frequency of significant bleeding appears to be similar. The authors describe a child on therapeutic doses of LMWH for a deep vein thrombosis who spontaneously developed an intramural hemorrhage in his small bowel, leading to infarction and a partial bowel resection. PMID- 16217262 TI - Primary malignant lymphoma of the central nervous system in an immunocompetent child: a case report. AB - Primary lymphoma of the central nervous system (PCNSL) is extremely rare, especially in childhood. A 9-year-old Japanese boy was diagnosed as having precursor-B cell-type lymphoblastic lymphoma, based on morphologic and immunocytochemical analysis of mononuclear cells in the cerebrospinal fluid and a positive reaction for terminal deoxynucleotidyl transferase (TdT), CD19, CD79a, and CD179b. After seven courses of chemotherapy and craniospinal radiotherapy, the patient is alive, well, and in continuous complete remission. Despite its rarity, PCNSL should be included in the differential diagnosis in the presence of symptoms of increased intracranial pressure and/or unusual imaging findings of the brain. PMID- 16217263 TI - Clonal monosomy 7 in a megakaryoblastic leukemia developed on the basis of Fanconi anemia. AB - A 13-year-old girl with a history of Fanconi anemia developed acute myeloid leukemia of the M7 subtype with a 45,XX,-7 karyotype, which is rare in M7 subtype. Treatment protocols were set up, but she died of sepsis and osteomyelitis during induction. PMID- 16217264 TI - Congenital hepatic fibrosis: a very uncommon cause of pancytopenia in children. AB - The disease presentation of autosomal recessive polycystic kidney disease (OMIM #263200, ARPKD) is highly variable and includes polycystic kidneys, pulmonary hypoplasia, and congenital hepatic fibrosis. The authors report an unusual case of ARPKD presenting with hepatosplenomegaly and cytopenia mimicking acute leukemia. PMID- 16217265 TI - Unique role of enalapril in the treatment of severe hypertension in a child with an unusual form of neuroblastoma. PMID- 16217270 TI - Renal cell carcinoma: urologists in a new era. PMID- 16217271 TI - Factors relating to residency training and medical student career choices in urology. PMID- 16217272 TI - Changing aspects in residency education. PMID- 16217273 TI - Quality of life after cystectomy and urinary diversion: an evidence based analysis. AB - PURPOSE: We critically examined the evidence supporting the widely accepted notion that patients undergoing continent urinary tract reconstruction after cystectomy experience superior quality of life outcomes than patients receiving a conduit. MATERIALS AND METHODS: Based on a comprehensive MEDLINE literature search we retrieved and evaluated all full-length articles published in the English, French, German, Italian and Spanish languages comparing conduit diversion with continent cutaneous diversion and/or orthotopic bladder substitution with respect to quality of life or similar concepts. All studies were rated according to the International Consultation on Urological Diseases modification of Oxford Center for Evidence-Based Medicine levels of evidence. RESULTS: The literature on quality of life after radical cystectomy for bladder cancer was rather extensive but generally of questionable quality. The main problems were flaws in the patient materials and methodologies used. To our knowledge not a single randomized, controlled study exists in the field. Because only few articles achieved a level of evidence better than III, the International Consultation on Urological Diseases rating system does not allow further differentiation among studies. Most studies showed that overall quality of life after cystectomy remained good in most patients irrespective of urinary diversion type. CONCLUSIONS: Existing studies are unable to prove that continent reconstruction after radical cystectomy is superior to conduit diversion. This review emphasizes the importance of performing well designed studies in the future. PMID- 16217274 TI - Incidence and management of gynecomastia in men treated for prostate cancer. AB - PURPOSE: Gynecomastia is a potentially treatment limiting adverse event in men receiving hormone therapy for prostate cancer. MATERIALS AND METHODS: In large, randomized, placebo controlled studies approximately 50% or more of patients with prostate cancer experienced gynecomastia due to multiple mechanisms. Although its severity was mostly reported as mild to moderate, gynecomastia was cited as the reason for most premature withdrawals from therapy. In patients with advanced forms of prostate cancer bilateral orchiectomy was associated with the lowest incidence of gynecomastia, followed by nonsteroidal antiandrogen therapy, diethylstilbestrol and estrogen in rank order. RESULTS: It is important that gynecomastia is well managed in patients with prostate cancer who want to proceed with hormone therapy. Patients should be assessed for the likely etiology of gynecomastia and preventive therapy or treatment for established gynecomastia should be instituted. Prophylactic radiotherapy has been shown to decrease the incidence of hormone induced gynecomastia by more than 50%. An alternative course of action, which may be more convenient for the patient, is the prophylactic use of tamoxifen. Tamoxifen may also mitigate or resolve gynecomastia during its early or proliferative phase. In severe long-standing gynecomastia surgery is warranted since medical therapies are less likely to succeed. Aromatase inhibitors and 4-hydroxytamoxifen are investigational. CONCLUSIONS: Gynecomastia is a significant problem in men undergoing hormonal therapy for prostate cancer. It requires prompt recognition, evaluation and management. PMID- 16217275 TI - Combined use of alpha-adrenergic and muscarinic antagonists for the treatment of voiding dysfunction. AB - PURPOSE: We provide an overview of the medical literature supporting the combined use of muscarinic and alpha-adrenergic antagonist therapy for the treatment of voiding dysfunction. MATERIALS AND METHODS: The MEDLINE database (1966 to 2004) of the United States National Library of Medicine was searched for pertinent studies. RESULTS: Although the mechanism of action of alpha-adrenergic antagonist therapy for voiding dysfunction has traditionally been assumed to be relaxation of the periurethral, prostatic and bladder neck smooth muscle, substantial evidence supports action at extraprostatic sites involved in micturition, including the bladder dome smooth muscle, peripheral ganglia, spinal cord and brain. Likewise the mechanism of action of anticholinergic therapy has been traditionally assumed to be inhibition of the M3 muscarinic receptor subtypes that mediate normal bladder contractions. However, M2 receptor mediates hypertrophied bladder contractions and there is evidence for an M2 component to the suprasacral control of voiding. CONCLUSIONS: Based on the physiology of alpha adrenergic and muscarinic receptors the inhibition of each one would be expected to be more beneficial than that of either alone because they would work on 2 components of detrusor function. Patients who would likely benefit from this combination therapy are men with lower urinary tract symptoms, women with urgency/frequency syndrome (overactive bladder), patients with uninhibited bladder contractions due to neurogenic bladder, and patients with pelvic pain and voiding symptoms, ie interstitial cystitis and chronic prostatitis/chronic pelvic pain syndrome. PMID- 16217276 TI - Renal cell and transitional cell carcinoma in a Japanese population undergoing maintenance dialysis. AB - PURPOSE: We verified differences in the incidence, clinical characteristics and outcomes between patients on chronic dialysis for end stage renal disease with renal cell carcinoma (RCC) and those with transitional cell carcinoma (TCC). MATERIALS AND METHODS: Data regarding RCC and TCC were reviewed in the medical records of 6,201 patients with end stage renal disease who underwent chronic dialysis between January 1990 and June 2003 in our 38 affiliated dialysis centers, and data were compared with those reported in Australia and New Zealand. RESULTS: Among the patients RCC developed in 38 (0.61%) and TCC developed in 16 (0.26%) during maintenance dialysis. The primary renal disease was chronic glomerulonephritis in patients with RCC (68.4%) and diabetic nephropathy in patients with TCC (43.8%, p = 0.002). Mean patient age at initiation of dialysis was 45 years for those with RCC and 63 for those with TCC (p < 0.001). Mean interval from dialysis induction to tumor diagnosis was 143 months for patients with RCC and 54 months for patients with TCC (p < 0.001). Of 38 RCCs 23 (60.5%) were incidentally detected by regular abdominal imaging examinations while painless gross hematuria was the cardinal symptom in 13 (81.2%) of 16 TCCs. Overall and cancer specific survivals after tumor diagnosis were significantly superior in patients with RCC compared to those with TCC (p = 0.0001 and p = 0.0003, respectively), and the cancer specific 5-year survival was 88.9% for RCC and 29.5% for TCC. In both cancers tumor stage significantly increased the risk of cancer specific death. Compared with patients from Australia and New Zealand, the incidence of RCC was higher and that of TCC was lower in our patients (p <0.001). CONCLUSIONS: In the Japanese population on dialysis RCC is more common than TCC. Since long-term dialysis is a risk factor for RCC, regular imaging examinations may have contributed to the favorable outcome of our patients on dialysis with RCC. In contrast, the unfavorable outcome of TCC suggests the need for effective diagnostic measures for early detection of TCC in patients on dialysis. PMID- 16217277 TI - Optical spectroscopy characteristics can differentiate benign and malignant renal tissues: a potentially useful modality. AB - PURPOSE: Promising results of optical signals have been reported in the literature for the diagnosis of Barrett's esophagus, oral cavity lesions, brain tumor margins, cervical intraepithelial neoplasia, skin cancer and bladder cancer. The potential usefulness of these techniques in renal tissues and neoplasms has not been described to date. This initial study examined the feasibility of using fluorescence and diffuse reflectance spectroscopy to differentiate between malignant and benign renal tissues. MATERIALS AND METHODS: An ex vivo study was conducted to identify optical characteristics of various renal tissue types. Pathologically confirmed benign and malignant renal samples were obtained from nephrectomy specimens from patients undergoing radical nephrectomy. Fluorescence and diffuse reflectance spectra were measured from benign and malignant renal tissues. RESULTS: All renal tissues, malignant or benign, contain 2 primary emission peaks-a strong one at approximately 285 nm excitation, approximately 340 nm emission (Peak A), and a weak one at approximately 340 nm excitation, approximately 460 nm emission (Peak B). Peak A of normal renal tissue typically locates at the shorter excitation wavelength region than that of malignant tissue. The intensity of Peak B from benign tissues tends to be greater than that from malignant renal tissues. Diffuse reflectance intensities from malignant renal tissues between 600 and 800 nm are markedly greater than those from normal renal tissue. Empirical discrimination algorithms developed based on selected fluorescence and diffuse reflectance spectral characteristics yields accurate differentiation between benign and malignant renal tissues. CONCLUSIONS: Highly accurate differentiation between normal human renal tissues and renal cell cancers is feasible using combined fluorescence and diffuse reflectance spectroscopy in an ex vivo setting. If successful in future clinical studies, optical spectroscopy could aid in margin detection and tissue discrimination while performing nephron sparing surgery. PMID- 16217278 TI - A scoring algorithm to predict survival for patients with metastatic clear cell renal cell carcinoma: a stratification tool for prospective clinical trials. AB - PURPOSE: We developed a clinically useful scoring algorithm to predict cancer specific survival for patients with clear cell metastatic renal cell carcinoma (RCC). MATERIALS AND METHODS: We studied 727 patients treated with radical nephrectomy for clear cell RCC from 1970 to 2000 who had distant metastases at nephrectomy (285) or in whom metastases subsequently developed (442). A scoring algorithm to predict cancer specific survival was developed using the regression coefficients from a Cox proportional hazards model. RESULTS: There were 606 deaths from clear cell RCC at a median of 1.0 years (range 0 to 14) following metastatic RCC. Constitutional symptoms at nephrectomy (+2), metastases to the bone (+2) or liver (+4), metastases in multiple simultaneous sites (+2), metastases at nephrectomy (+1) or within 2 years of nephrectomy (+3), complete resection of all metastatic sites (-5), tumor thrombus level I to IV (+3), and the primary pathological features of nuclear grade 4 (+3) and histological tumor necrosis (+2) were significantly associated with death from RCC. All patients started with a score of 0 and points were added or subtracted as indicated in parentheses. Cancer specific survival rates at 1 year were 85.1%, 72.1%, 58.8%, 39.0%, and 25.1%, respectively, for patients with scores of -5 to -1, scores of 0 to 2, scores of 3 to 6, scores of 7 or 8, and scores of 9 or more. CONCLUSIONS: This scoring algorithm can be used to predict cancer specific survival for patients with metastatic clear cell RCC. PMID- 16217279 TI - Long-term outcome of transcatheter embolization of renal angiomyolipomas due to tuberous sclerosis complex. AB - PURPOSE: Complications from renal angiomyolipomas (AMLs) are common in patients with tuberous sclerosis complex (TSC) and tumors greater than 4 cm are more likely to cause symptoms. AMLs are the most common cause of death in adults with TSC. We present our long-term experience with transcatheter tumor embolization as a definitive treatment for AMLs due to TSC. MATERIALS AND METHODS: A total of 16 patients with TSC between 7.5 and 47.2 years old with symptomatic or large (4 to 21 cm) AMLs underwent embolization. Followup consisted of periodic physician visits or telephone contacts and renal imaging. RESULTS: The 16 patients underwent 18 treatment sessions to embolize 27 tumors. There were no intraoperative complications. The post-embolization syndrome occurred in 11 individuals but all responded to medical management. Two individuals had an arterial aneurysm within a tumor. The AML size decreased in the 13 patients who were imaged 3 months after treatment, and the 7 patients who were imaged 3 to 9 years after treatment have shown no tumor regrowth. No renal failure or hemorrhage has developed in patients following embolization. CONCLUSIONS: Transcatheter embolization of symptomatic or large AMLs due to TSC prevents hemorrhage and renal loss. The treatment is minimally invasive, preserves renal function, and can be performed multiple times. All of the patients who underwent followup renal imaging after embolization showed decreased AML size, and none of the 16 patients has developed renal loss or renal insufficiency in these individuals. Embolization should be considered the initial treatment of choice for large or symptomatic AMLs. PMID- 16217280 TI - Contribution of the prostate limits the usefulness of survivin for the detection of bladder cancer. AB - PURPOSE: We determined if urinary survivin is influenced by the prostate and analyzed survivin levels in a healthy control population. MATERIALS AND METHODS: This was a blinded, retrospective analysis of frozen urine samples. Three groups were designated. Group 1 patients were diagnosed with prostate cancer and samples were collected immediately prior to radical retropubic prostatectomy. Group 2 patients had already undergone radical retropubic prostatectomy at least 3 months prior to providing a urine sample. Group 3 comprised healthy controls. Groups 1 and 2 patients were recruited at an institutional practice. Group 3 patients were recruited from the local community. RESULTS: The frequency of high survivin scores was much lower in patients who did not have a prostate (p < 0.0002). The survivin score was 2 in 35% and 44% of the patients in groups 1 and 3, respectively, while only 9% of those in group 2 had a survivin score of 2 (p < 0.0002). Of healthy controls 50% had a false-positive survivin score. Urinary survivin predicted prostate cancer poorly with 52% sensitivity and 50% specificity, and it did not predict any biological features of prostate cancer. CONCLUSIONS: Urinary survivin markedly decreases after prostatectomy. There is a 50% false-positive rate when using urinary survivin in controls. These 2 features make urinary survivin a poor bladder cancer biomarker. PMID- 16217281 TI - Serum CYFRA 21-1 in patients with invasive bladder cancer and its relevance as a tumor marker during chemotherapy. AB - PURPOSE: Previous studies have shown that serum levels of the degradation products of cytokeratins could be used as surrogate markers in the diagnosis and followup of patients with solid tumors, including tumors of the bladder. MATERIALS AND METHODS: The soluble cytokeratin 19 fragment CYFRA 21-1 was measured by solid phase radioimmunoassay in the serum of 142 patients with invasive transitional cell cancer of the bladder. Of the patients 56 had clinical stage I to III locally confined disease (T1-4aN0M0) and 86 had stage IV metastatic disease with lymph node and/or distant metastases. A control group consisted of 33 healthy volunteers. In a subgroup of 49 patients with metastatic disease receiving combined platinum based chemotherapy serum CYFRA 21-1 was determined prior to the initiation of therapy and after the documentation of response. RESULTS: Abnormal CYFRA 21-1 was observed in 7% of patients with locally invasive disease and in 66% of those with metastatic disease (p < 0.0001). There was no correlation of CYFRA 21-1 with tumor differentiation. Patients with abnormal CYFRA 21-1 showed statistically significant worse median overall survival. Moreover, in the subgroup of patients with metastatic disease receiving chemotherapy CYFRA 21-1 levels correlated with the response to treatment. CONCLUSIONS: Patients with transitional cell cancer of the bladder with evidence of distant metastases showed a significant increase in serum CYFRA 21-1. During chemotherapy CYFRA 21-1 appears to be a potentially sensitive and useful indicator for monitoring treatment response. PMID- 16217284 TI - Percutaneous CT-guided radiofrequency ablation of renal neoplasms: factors influencing success. PMID- 16217283 TI - Long-term survival of patients with unilateral sporadic multifocal renal cell carcinoma according to histologic subtype compared with patients with solitary tumors after radical nephrectomy. PMID- 16217282 TI - Treatment delay and prognosis in invasive bladder cancer. AB - PURPOSE: We studied treatment delay, and the impact on disease specific survival and stage progression in a series of patients who had undergone cystectomy. MATERIALS AND METHODS: All 141 patients underwent radical cystectomy between 1990 and 1997 due to locally advanced bladder cancer. Treatment delay was defined as time from pathological confirmation of invasive disease to performance of cystectomy, and was registered retrospectively from the patient charts. Two patients received neoadjuvant chemotherapy and were excluded from further analyses. Followup continued until April 2003 with death due to bladder cancer as the end point. Causes of death were retrieved from the Swedish Cause of Death Registry. RESULTS: The median treatment delay was 49 days, but was significantly longer for the 71 cases who were referred from other hospitals (63 vs 41 days, p < 0.001). Treatment delay did not influence cumulative incidence of death from bladder cancer. Considering all cases, there was no significant correlation between treatment delay and stage progression. For clinical stage T2 tumors, median treatment delay was 76 days among patients with stage progression compared to 41 and 48 days for those with stage regression and stage equivalence, respectively (p = 0.20). CONCLUSIONS: Treatment delay was not found to influence disease specific survival in the present study. Furthermore, treatment delay was not significantly longer in cases that progressed compared to those with equal or lower pathological stage in the cystectomy specimen. PMID- 16217285 TI - The influence of intravesical therapy on progression of superficial transitional cell carcinoma of the bladder. A metaanalytic comparison of chemotherapy versus bacilli Calmette-Guerin immunotherapy. PMID- 16217286 TI - Weekly paclitaxel and gemcitabine in advanced transitional-cell carcinoma of the urothelium: a Phase II Hoosier Oncology Group Study. PMID- 16217287 TI - Comparison of the incidence of latent prostate cancer detected at autopsy before and after the prostate specific antigen era. AB - PURPOSE: Most data regarding the prevalence of latent prostate cancer found only at autopsy are from old reports. To determine if significant differences exist in the prevalence of latent prostate cancer between periods before and after the advent of screening for prostate cancer, we compared 2 groups of men undergoing autopsy during the 2 periods. MATERIALS AND METHODS: Our institutional autopsy record database was searched to identify all men found to have prostate cancer before or after death between 1955 and 1960 (total 3,307 men and 1,578 men older than 40 years), and between 1991 and 2001 (total 2,938 men, 1,380 men older than 40 years). We calculated the age based incidence of latent prostate cancer detected only at autopsy in an at risk population of men (older than 40 years). We also compared Gleason grade distribution and proportion of stage cT3 or greater cancers between the 2 periods. RESULTS: Between 1955 and 1960 the prevalence of latent prostate cancer detected only at autopsy in men older than 40 years was 4.8% compared to 1.2% (p < 0.0001) between 1991 and 2001. A significant decrease in the prevalence of latent, autopsy detected cancers was observed in men 70 to 89 years old at death. Autopsy detected cancers were found to be grossly invading adjacent structures (stage cT3 or greater) in 17 of 76 (22%) cancers discovered between 1955 and 1960, while none of the latent prostate cancers found in the 1991 to 2001 period were found to extend grossly beyond the prostate. CONCLUSIONS: Autopsy rates are decreasing at our institution. With the more widespread use of screening, the prevalence of latent prostate cancer has decreased 3-fold. The decrease in the prevalence of latent prostate cancer is especially dramatic in men older than 70 years. Further study will determine the significance of many of the tumors currently detected clinically, which may have been latent and found at autopsy if not for screening. PMID- 16217288 TI - The independent impact of extended pattern biopsy on prostate cancer stage migration. AB - PURPOSE: There are many factors impacting stage migration for prostate cancer. The number of prostate core biopsies is known to increase detection of prostate cancers. It is still unknown whether the number of biopsies is an independent predictor of tumor size. This is important as a number of studies show that tumor volume is an independent predictor of cancer progression. MATERIALS AND METHODS: Using the University of California, San Francisco Urologic Oncology database, a retrospective review of 378 patients undergoing radical prostatectomy by a single surgeon during 2000 to 2003 was performed. Patient and tumor specific variables including age, prostate specific antigen (PSA), number of biopsies, biopsy Gleason grade, tumor volume in the surgical specimen and surgical specimen tumor grade were studied. Univariate and multivariate statistical methods including multiple and logistic regression were used to characterize patients by the number of biopsy cores. Tests of significance to identify predictors of tumor size were based on the partial F statistic and the likelihood ratio test. RESULTS: A total of 317 eligible patients were studied, of whom 119 had 6 biopsies and 198 had more than 6 biopsies. The 2 groups of patients were evenly matched in terms of age, PSA and Gleason sum, with no statistically significant differences observed. On univariate analysis, mean tumor volume was larger for patients receiving 6 core biopsies vs greater than 6 core biopsies (3.85 vs 2.04 cc, p = 0.0009). Additionally, statistically significant differences were observed when comparing median tumor volumes, as well as excluding extremely large volume tumors. On multivariate analysis the number of biopsies performed (6 vs more than 6), was an independent predictor of tumor size (p = 0.006), controlling for primary Gleason score, Gleason sum, PSA as a continuous or categorical variable, year of biopsy and year of surgery. CONCLUSIONS: The use of extended pattern prostate biopsy templates results in the detection of smaller volume prostate cancers, independent of PSA and Gleason grade. These biopsy templates have contributed to the downward stage migration of prostate cancer detection and may possibly contribute to the risk of over detection. PMID- 16217289 TI - Management of prostate cancer in China: a multicenter report of 6 institutions. AB - PURPOSE: In China the incidence of prostate cancer (PCa) is low and sparse data are available regarding its management. We analyzed the management of PCa at 6 Chinese urological institutions. MATERIALS AND METHODS: A retrospective analysis was performed of 431 consecutive patients treated for PCa at 6 Chinese institutions, including 5 in the divisions of Shanghai and 1 in the province of Chongqing, between January 2000 and December 2004. Tumor characteristics, therapeutic options and patient outcomes were recorded. RESULTS: At diagnosis median patient age was 72 years and median prostate specific antigen was 46.1 ng/ml. Most PCa cases were revealed by urinary symptoms (75.9%) or bone pain (12.8%). PCa was palpable on digital rectal examination in 74% of cases. At least 44 patients (10.2%) had metastases to lymph nodes at diagnosis and 112 (26%) had bone metastases. A total of 236 patients underwent bilateral orchiectomy and 100 received medical hormone therapy, which in 75% consisted of antiandrogen alone. At a median followup of 16.8 months 60% of these patients experienced biological recurrence. Radical prostatectomy was performed in 24 patients as monotherapy or in combination with bilateral orchiectomy. No patient with clinically localized PCa experienced biological recurrence after radical prostatectomy. CONCLUSIONS: The management of PCa in China differs from that in Western countries. To date surgical castration represents standard treatment. Screening detection of PCa could help detect earlier stage tumors and improve the outcome in patients. PMID- 16217290 TI - Obesity and capsular incision at the time of open retropubic radical prostatectomy. AB - PURPOSE: The goal of radical prostatectomy (RP) is complete removal of the intact prostate. Obese men can represent a technical challenge. However, to our knowledge objective data linking obesity with technically inferior surgery are lacking. Therefore, we examined the association between body mass index (BMI) and capsular incision at RP as a surrogate of a poor technical operation in men treated for prostate cancer by several high volume surgeons at a center of excellence. MATERIALS AND METHODS: The study population consisted of 7,027 men treated with anatomical retropubic RP between 1996 and 2004 by 7 high volume surgeons. We evaluated the association between BMI and capsular incision using logistic regression, adjusting for clinical and pathological variables, and for the surgeon. RESULTS: Overall capsular incision was noted in 4.6% of all RP specimens. After adjustment for preoperative prostate specific antigen, patient race, height, year of surgery, clinical stage, pathological Gleason sum, prostate weight, extraprostatic extension and seminal vesicle invasion increased BMI was associated with increased odds of capsular incision (p trend = 0.005). After further adjustment for surgeon mild obesity was associated with 30% increased odds of capsular incision (OR 1.30, 95% CI 0.92 to 1.83), while moderate and severe obesity was associated with 57% increased odds of capsular incision (OR 1.57, 95% CI 0.82 to 3.00) relative to normal weight men (p trend = 0.06). CONCLUSIONS: In a study of more than 7,000 men treated by 7 experienced surgeons BMI was positively related to capsular incision. This suggests that open retropubic RP is technically more difficult in obese men, which results in a greater likelihood of a less than technically ideal operation. Although this may be predicted to have a negative impact on disease-free survival outcomes in obese men, it is unlikely to alone explain the worse outcomes in obese men noted in previous RP series. PMID- 16217291 TI - Androgen deprivation use with external beam radiation for prostate cancer: results from CaPSURE. AB - PURPOSE: Recent data support the role of androgen deprivation in men undergoing external beam radiotherapy for prostate cancer. The benefits of neoadjuvant, concurrent or adjuvant treatment have been limited to men at intermediate and high risk. We examined the patterns and predictors of androgen deprivation in men undergoing external beam radiation therapy in CaPSURE. MATERIALS AND METHODS: CaPSURE is an observational, longitudinal disease registry, from which 932 men met study inclusion criteria. Androgen deprivation was classified as neoadjuvant within 9 months of radiation or adjuvant-from the start of radiation to 6 months after completion. Time trends in androgen deprivation as well factors associated with combined therapy were elucidated using multivariate analyses. RESULTS: In this study 40%, 39% and 21% of men could be categorized into high, intermediate and low risk groups, respectively. Overall 42% and 33% of patients received neoadjuvant and adjuvant androgen deprivation therapy, respectively. Between 1997 and 2002 neoadjuvant hormone use increased significantly in all risk groups, including patients at low risk. On multivariate analyses only the year of diagnosis and clinical risk group were associated with receiving androgen deprivation with radiation. CONCLUSIONS: A significant increase in combined androgen deprivation and external radiation was observed in the last decade in men with intermediate and high risk disease. Nevertheless, more widespread acceptance is necessary since a substantial minority continue to receive radiation alone. Many patients with low risk disease that is amenable to radiation monotherapy also receive androgen deprivation. No clinical or sociodemographic features predicted the use of androgen deprivation with external radiation. PMID- 16217292 TI - Phase II trial of tesmilifene plus mitoxantrone and prednisone for hormone refractory prostate cancer: high subjective and objective response in patients with symptomatic metastases. AB - PURPOSE: Symptomatic, hormone refractory prostate cancer (HRCAP) is a major cause of morbidity with a median survival of less than 12 months and a 2-year survival of only up to 10% in most series. Mitoxantrone has been approved by the Food and Drug Administration for HRCAP. Preliminary data suggest that DPPE (N,N-diethyl-2 [4-(phenylmethyl) phenoxy]-ethanamine) or tesmilifene modulates cytotoxics to enhance the anticancer effect. In this phase II trial we assessed whether there is sufficient evidence of enhanced efficacy of DPPE and mitoxantrone to lead to a phase III clinical trial. MATERIALS AND METHODS: A total of 29 patients with a median age of 73 years, of whom 10% were older than 80 years, with progressive HRCAP received 5.3 mg/kg DPPE intravenously every 3 weeks, 12 mg/m mitoxantrone intravenously every weeks and 5 mg prednisone orally twice daily. All patients had pain at presentation, while 97% had bone metastases, 10% had liver metastases and 17% had lung metastases. Median prostate specific antigen (PSA) was 210 ng/ml (IQR 77 to 430). RESULTS: Of the patients 75% had some pain improvement, 66% had decreased analgesia, 59% had a PSA decrease of 50% or greater and 45% had a PSA decrease of 75% or greater. Actual (not actuarial) 2-year survival was 21%. CONCLUSIONS: Despite major limitations of historical comparison the PSA decrease and decreased symptoms with DPPE-mitoxantrone-prednisone compare favorably to those of mitoxantrone-prednisone and docetaxel-estramustine in the literature. The 2-year survival rate of 21% mandates further assessment. This will be tested in a phase III Southwest Oncology Group trial. PMID- 16217293 TI - Tissue microarray analysis of hMSH2 expression predicts outcome in men with prostate cancer. AB - PURPOSE: Mismatch repair genes are responsible for the coordinated correction of misincorporated nucleotides formed during DNA replication. Mismatch repair expression is altered in a subset of prostate cancers (PCs) and a recent study suggested that time to biochemical recurrence following prostatectomy correlated with the degree of hMSH2 immunohistochemical staining. We compared hMSH2 expression and survival in clinically organ confined PC. MATERIALS AND METHODS: A prostate tissue microarray was constructed using 243 specimens from patients who underwent radical prostatectomy with extended lymph node dissection for clinically organ confined PC with up to 12 years of followup. Immunohistochemistry was performed with anti-human MSH2 monoclonal antibody. Three independent observers evaluated hMSH2 expression on a scale of 0 to 4. Low expression was defined as a score of less than 2 and high expression was defined as a score of 2 or higher. Statistical analysis used the Fisher exact test, and Goodman and Kruskal gamma coefficient. RESULTS: Higher Gleason score significantly correlated with higher hMSH2 expression (p < 0.0002). Low hMSH2 expression correlated with increased overall, disease-free and biochemical disease-free survival (all p < 0.01). Analysis comparing low vs high hMSH2 expression was significant with respect to overall (p = 0.0004), disease-free (p = 0.005) and biochemical disease-free (p = 0.0177) survival. CONCLUSIONS: hMSH2 is differentially expressed in malignant prostate tissue and hMSH2 immunohistochemical staining intensity correlates with Gleason score, overall and disease-free survival. Taken together our results suggest that hMSH2 expression may be a useful prognostic biomarker for outcome in men with clinically organ confined PC. PMID- 16217294 TI - Increased incidence of testicular cancer in men presenting with infertility and abnormal semen analysis. AB - PURPOSE: We determined the standardized incidence ratio of testicular cancer in infertile men presenting with an abnormal semen analysis compared to the general population. MATERIALS AND METHODS: The charts from more than 3,800 men presenting with infertility and abnormal semen analysis during a 10-year period were retrospectively reviewed. The incidence of testicular tumors diagnosed in this group was compared to that of race and age matched controls during the same period from the general population (as reported by the Surveillance, Epidemiology and End Results [SEER] database). RESULTS: Of 3,847 men 10 (0.3%) with infertility and abnormal semen analysis were diagnosed with testicular tumors. Mean patient age was 32.6 years (range 25 to 52) and all 10 men were diagnosed with a seminomatous germ cell tumor. Two men had a history of cryptorchidism while the remaining 8 had no identifiable risk factors for testicular cancer. The SEER database reported an incidence of 10.6 cases of testicular cancer (95% CI 10.3-10.8) per 100,000 men of similar age group and racial composition during the same period. The standardized incidence ratio of testicular cancer was 22.9 (95% CI 22.4-23.5) when comparing our infertile group to the control population. Exclusion from analysis of the 2 patients with a history of cryptorchidism decreased the standardized incidence ratio to 18.3 (95% CI 18.0-18.8). CONCLUSIONS: Infertile men with abnormal semen analyses have a 20-fold greater incidence of testicular cancer compared to the general population. Patients and physicians should be aware that one of the causes of infertility could be cancer, particularly testicular cancer. PMID- 16217295 TI - Effects of long-term vitamin E supplementation on cardiovascular events and cancer: a randomized controlled trial. PMID- 16217296 TI - Screening by prostate-specific antigen and digital rectal examination in relation to prostate cancer mortality. A case-control study. PMID- 16217297 TI - Serum levels of phytanic acid are associated with prostate cancer risk. PMID- 16217298 TI - Observed effect of age and body mass index on total and complexed PSA: analysis from a National Screening Program. PMID- 16217299 TI - The association of body mass index and prostate-specific antigen in a population based study. PMID- 16217300 TI - Gene expression profiling of early- and late-relapse nonseminomatous germ cell tumor and primitive neuroectodermal tumor of the testis. PMID- 16217301 TI - Renal pedicle lymphatic disconnection for chyluria via retroperitoneoscopy and open surgery: report of 53 cases with followup. AB - PURPOSE: We present our experience with retroperitoneoscopic renal pedicle lymphatic disconnection. We compared the clinical efficacy of this treatment for chyluria with that of open surgery. MATERIALS AND METHODS: From January 1998 to June 2004, 53 patients (55 renal units) with chyluria underwent renal pedicle lymphatic disconnection via the retroperitoneoscopic and conventional open approaches. The diagnosis of chyluria was confirmed by the ether test and the side of chylous reflux was determined by cystoscopy. Operative time, intraoperative blood loss, postoperative intestinal recovery and hospital stay were evaluated. Increases in hemoglobin and serum albumin were compared before and after surgery during followup. RESULTS: Retroperitoneoscopic renal pedicle lymphatic disconnection or open surgery was performed successfully in all patients. In terms of operative time, intraoperative blood loss, postoperative intestinal recovery and hospital stay retroperitoneoscopy was superior to conventional open surgery. During retroperitoneoscopy the inferior vena cava was injured in 1 case but repaired successfully by laparoscopy without conversion to open surgery. Postoperative gross hematuria in 1 case disappeared 4 days later. In the open surgery group the renal segmental artery was inadvertently injured in 1 case and anastomosis was performed successfully. Wound healing was delayed in 1 case due to hypoalbuminemia. Recurrence developed in 2 patients during the 6 to 84-month followup. CONCLUSIONS: Retroperitoneoscopic renal pedicle lymphatic disconnection for chyluria has the advantages of minimal invasion and rapid recovery compared with open surgery. PMID- 16217302 TI - Menstrual cycle affects bladder pain sensation in subjects with interstitial cystitis. AB - PURPOSE: Using psychophysical methods we compared the effect of the menstrual cycle on bladder sensation in subjects with the diagnosis of interstitial cystitis (IC) and in controls. MATERIALS AND METHODS: Female participants with normal menstrual cycles, including 7 with IC and 8 healthy controls, were recruited into this study. They completed daily diaries related to bladder pain and other body pain, and tracked daily micturition frequency. In a subset formal psychophysical testing of thermal and ischemic pain was performed at 2 times of the menstrual cycle, corresponding to the luteal and follicular phases. Cystometrograms were performed at the same time. RESULTS: Subjects with IC had higher pain scores and frequency than controls throughout the entire menstrual cycle. Pain scores were highest in the perimenstrual period in subjects with IC and controls. Micturition frequency was highest in the perimenstrual period in subjects with IC. Cystometric evaluation of a first need to void and the evocation of bladder pain demonstrated that lower intravesical volume and pressure were necessary to evoke bladder pain during the follicular period than during the luteal period in subjects with IC, although there was no effect on the first need to void. CONCLUSIONS: These findings are consistent with clinical lore that suggests a perimenstrual flare in pain in subjects with IC. To our knowledge it also demonstrates for the first time a menstrual cycle effect on bladder sensory function in subjects with IC. This suggests a potential role of gonadal hormones on bladder sensory processing and, therefore, a potential role for hormonal modulation as a therapeutic modality in this patient population. PMID- 16217303 TI - Long-term results of amitriptyline treatment for interstitial cystitis. AB - PURPOSE: We performed a prospective, open label study to examine the safety and efficacy of the long-term administration of the tricyclic antidepressant amitriptyline in patients with interstitial cystitis (IC). MATERIALS AND METHODS: A total of 94 patients were stratified into 2 groups, namely a National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) group of those who fulfilled NIDDK criteria for IC and a nonNIDDK group of those who presented with characteristic IC symptoms but met at least 1 NIDDK exclusion criterion. Amitriptyline was received strictly at bedtime following an established self titration protocol without a limitation of the maximum daily dose. Patients reporting improvement in a global response assessment questionnaire were considered treatment responders. Further efficacy measures were changes in pain and urgency, functional bladder capacity and frequency. Changes in the O'Leary Sant IC index and rating of overall satisfaction with the therapeutic outcome were also reported. RESULTS: Mean study followup +/- SD was 19.0 +/- 12.5 months. The response rate was 64% (60 patients). The overall mean dose was 55 mg (range 12.5 to 150). Side effects occurred in 79 patients (84%), including dry mouth in 79% and weight gain in 59%. Patient overall satisfaction with the therapeutic result was excellent or good in 43 (46%). The dropout rate was 31% (29 patients) after a mean treatment period of 6 weeks at a mean dose of 70 mg. Nonresponse to treatment was the primary reason for dropout in all cases, while side effects contributed to dropout in 25 (86%). Improvement in the various IC symptoms was statistically significant compared with baseline. CONCLUSIONS: Long-term administration of amitriptyline is a feasible, safe and effective treatment for IC, provided that the drug is used judiciously to minimize adverse effects. The therapeutic response to amitriptyline was uniformly observed in patients fulfilling NIDDK criteria and in those with the pure clinical diagnosis of IC. PMID- 16217304 TI - Economic evaluation of an updated guideline for the empiric treatment of uncomplicated urinary tract infection in women. PMID- 16217305 TI - Risk factors associated with acute pyelonephritis in healthy women. PMID- 16217306 TI - Predictors of quality of life and pain in chronic prostatitis/chronic pelvic pain syndrome: findings from the National Institutes of Health Chronic Prostatitis Cohort Study. PMID- 16217307 TI - The prevalence of urinary tract infections and sexually transmitted disease in women with symptoms of a simple urinary tract infection stratified by low colony count criteria. PMID- 16217308 TI - Lactobacillus sepsis associated with probiotic therapy. PMID- 16217309 TI - Intracellular bacterial communities of uropathogenic Escherichia coli in urinary tract pathogenesis. PMID- 16217310 TI - Host subversion by formation of intracellular bacterial communities in the urinary tract. PMID- 16217311 TI - Jejunoileal bypass reversal: effect on renal function, metabolic parameters and stone formation. AB - PURPOSE: While the effect of jejunoileal bypass (JIB) reversal has been well studied regarding hepatic function, there is little information regarding the effect of reversal on renal function and even less data regarding the metabolic urinary stone environment. We evaluated the results of JIB reversal on renal function, the urinary stone milieu and the clinical development of recurrent calculi in affected patients. MATERIALS AND METHODS: From 1995 to 2003, 4 female patients with a mean age of 48.2 years underwent JIB reversal primarily for refractory stone disease. The clinical and metabolic courses prior to and following bypass reversal were reviewed specifically to evaluate renal function, serum and urinary metabolic stone profiles, and clinical stone formation. RESULTS: At initial presentation following JIB all 4 patients had significantly increased 24-hour urinary oxalate (range 80 to 160 mg, mean 112.5, normal less than 50) and significantly low 24-hour urinary citrate (range 5 to 62 mg, mean 21.5, normal greater than 320). Following reversal 24-hour urinary oxalate normalized to between 31 and 36 mg (mean 33.75). However, 24-hour urinary citrate continued to be low (range 215 to 248 mg, mean 226.5). After JIB reversal all 4 patients continued to have new stones until the commencement of urinary alkalization, following which only 1 had 1 calculus, which occurred 47 months after reversal. After JIB mean serum creatinine was 1.48 mg/dl (range 0.8 to 1.9) and mean urinary creatinine excretion was 0.91 mg per hour (range 0.69 to 1.15). After JIB reversal mean serum creatinine was 1.28 mg/dl (range 0.6 to 2.0) and mean urinary creatinine excretion was 1.0 mg per hour (range 0.85 to 1.10). CONCLUSIONS: JIB reversal normalizes 24-hour urinary oxalate. While urinary citrate improves, it continues to be low and such patients are at high risk for recurrent stone formation. However, in this setting appropriate replacement therapy has a significant and positive impact on that propensity. PMID- 16217312 TI - The safety and efficacy of direct trocar insertion with elevation of the rectus sheath instead of the skin for pneumoperitoneum. PMID- 16217313 TI - Ureteric access with flexible ureteroscopes: effect of the size of the ureteroscope. PMID- 16217314 TI - Comparison of needlescopic appendectomy versus conventional laparoscopic appendectomy. A randomized controlled trial. PMID- 16217315 TI - The heilbronn laparoscopic training program for laparoscopic suturing: concept and validation. PMID- 16217316 TI - Laparoscopic ureterocalicostomy: development of a technique simplified by application of Nitinol clips and a wet monopolar electrosurgery device. PMID- 16217317 TI - Endoureterotomy by intraluminal invagination for nonmalignant ureterointestinal anastomotic strictures: description of a new surgical technique and long-term followup. AB - PURPOSE: We describe a new surgical endoscopic technique for nonmalignant ureterointestinal anastomotic strictures. This procedure involving endoureterotomy by intraluminal invagination (the Lovaco technique) is performed by adopting a combined percutaneous antegrade and endoscopic retrograde approach. The results obtained by this technique are reviewed with long-term followup. MATERIALS AND METHODS: A total of 25 ureterointestinal anastomotic strictures were subjected to endoureterotomy by intraluminal invagination, including 12 left, 7 right and 3 bilateral cases. Surgical success was defined by radiological improvement and/or the ability to recover normal activity in the absence of flank pain, infection, or the need for ureteral stents or nephrostomy tubes. RESULTS: At a median followup of 51 months (range 2 to 145) the success rate for endoureterotomy by intraluminal invagination was 80% (20 of 25 ureterointestinal anastomotic strictures). No complications were recorded in the patients following endoureterotomy. CONCLUSIONS: This new endoureterotomy technique for ureterointestinal strictures following urinary diversion can be applied to any type of urinary diversion. It allows direct visualization of the stricture and stricture tissue biopsy. Intraluminal invagination makes it possible to increase the distance between the stricture, and the retroperitoneal vessels and bowels. The technique provides the control required to ensure full-thickness and full length stricture incision. The success rate is high and it persists after long term followup. PMID- 16217318 TI - Improved hemostasis during laparoscopic partial nephrectomy using gelatin matrix thrombin sealant. PMID- 16217319 TI - End-to-end urethroplasty: long-term results. PMID- 16217320 TI - Tissue-engineered buccal mucosa for substitution urethroplasty. PMID- 16217321 TI - Hand-assisted laparoscopic cystoprostatectomy and urinary diversion. PMID- 16217322 TI - CT urography of urinary diversions with enhanced CT digital radiography: preliminary experience. PMID- 16217323 TI - Mucus production after transposition of intestinal segments into the urinary tract. PMID- 16217324 TI - Caveats when interpreting intravenous urograms following ileal orthotopic bladder substitution. PMID- 16217325 TI - Brain control of normal and overactive bladder. AB - PURPOSE: Bladder control problems are common but their cause is often unclear. Many investigators have sought causes in the lower urinary tract, but fewer in the supraspinal control system. We have used functional magnetic resonance imaging (fMRI) to determine brain responses to bladder filling in subjects with normal and with poor bladder control (detrusor overactivity). MATERIALS AND METHODS: Cerebral responses to bladder infusion were recorded in 1 male and 11 females without overt neurological abnormality, aged 26 to 83 years. Six had good bladder control and 6 had poor control on prior urodynamics. fMRI was performed while repeatedly infusing and withdrawing liquid into and out of the bladder, and monitoring intravesical pressure. Measurements were made at small and large bladder volumes. RESULTS: fMRI detected activation of many brain regions involved in bladder control, including periaqueductal gray, thalamus, insula, dorsal anterior cingulate, and ventromedial cerebellum. Orbitofrontal cortex, pontine micturition center and preoptic hypothalamus were visible in subgroup analyses. Activations outweighed deactivations and responses became stronger at large bladder volumes. Among subjects with good control, this strengthening of response was prominent in the orbitofrontal cortex. Among those with poor control cortical responses were exaggerated at larger bladder volumes, except in the orbitofrontal cortex, which remained weakly activated. This difference was not due to concurrent detrusor activity. CONCLUSIONS: Poor bladder control is specifically associated with inadequate activation of orbitofrontal cortex. Clinically, frontal cortical lesions cause bladder control problems. This study suggests a similar neurophysiological basis for poor bladder control in the absence of overt neurological lesion. PMID- 16217326 TI - Factors that distinguish continent from incontinent older adults with detrusor overactivity. AB - PURPOSE: In older adults detrusor overactivity (DO) is almost as common in continent individuals as in those with urge incontinence (UUI). Thus, UUI likely reflects the contribution of additional factors. We postulated that of functionally independent individuals in whom transient causes were excluded those in whom DO was accompanied by UUI would be more likely to have smaller functional bladder capacity, less warning and less ability to avert urine loss in the face of DO. MATERIALS AND METHODS: While blinded to continence status, we reviewed the records of all 52 cognitively intact and nonobstructed volunteers older than 65 years who had DO associated with urgency during urodynamic testing and had been asked to try to prevent leakage for 2 minutes. Of these individuals 31 were continent at home. RESULTS: In patients with UI mean functional bladder capacity +/- SD was smaller (313 +/- 153 vs 390 +/- 178 ml, p = 0.06) and warning was briefer (87 +/- 81 vs 120 +/- 116 ml), although the association with warning was less impressive (p = 0.14). The ability to forestall leakage for at least 2 minutes correlated strongly with being continent at home (p < 0.002). For continence detrusor suppression at the onset of DO was more important than sphincter contraction. CONCLUSIONS: Urge incontinence in older adults involves more than simply the presence of DO. Physical function, cognitive function and medications are already known to be important for continence. Our data suggest that functional bladder capacity and the ability to suppress involuntary contraction (and possibly warning) are also potentially important covariates. These factors may represent additional potential targets for behavioral and pharmacological therapy. PMID- 16217327 TI - Effects of botulinum toxin B on refractory detrusor overactivity: a randomized, double-blind, placebo controlled, crossover trial. AB - PURPOSE: Open, observational studies of intradetrusor injections of botulinum toxin for detrusor overactivity have reported beneficial effects. We tested the efficacy and safety of botulinum toxin B for the treatment of the overactive bladder in a randomized, double-blind, placebo controlled crossover trial. MATERIALS AND METHODS: A total of 20 patients 18 to 80 years old with detrusor overactivity unresponsive to oral antimuscarinic agents participated in the study. They were injected with either placebo (20 ml normal saline) or botulinum toxin B (5,000 IU diluted up to 20 ml) intravesically in a day case setting. After 6 weeks the treatments were crossed over without washout in line with previous findings. The primary outcome was the paired difference in change in average voided volumes. Frequency, incontinence episodes and paired differences in quality of life measured by the King's Health Questionnaire were the secondary outcome measures. RESULTS: The Wilcoxon signed ranks test was used to test the paired difference in change between treatment phases. Little carryover was noted in the second arm placebo and the placebo data from both arms were included in analysis. There were clinically statistically significant paired differences in the change in average voided volume, urinary frequency and episodes of incontinence between active treatment and placebo (average voided volume: 95% CI difference 16, 122; Z2.5; p = 0.012/weekly frequency: 95% CI -21, -1; Z2.1, p=0.033/weekly incontinence: 95% CI -26, -7; Z3.3; p = 0.001). There were similarly significant paired differences in the change in quality of life affecting 5 domains of the King's Health Questionnaire. CONCLUSIONS: This double blind, placebo controlled, crossover study provides evidence of the efficacy of botulinum toxin B in the treatment of overactive bladder. Autonomic side effects were observed in 4 patients. The short duration of action will presumably limit the use to patients who have experienced tachyphylaxis with botulinum toxin A. PMID- 16217328 TI - Long-term urinary continence rates after simple sling incision for relief of urinary retention following fascia lata pubovaginal slings. AB - PURPOSE: We examined long-term urinary continence rates in patients after midline simple sling incision for urinary retention following suburethral fascia lata slings. MATERIALS AND METHODS: A retrospective review was completed of 13 women undergoing a simple sling incision for catheter dependent obstruction after suburethral sling surgery more than 4 years previously. Urinary continence was evaluated by use of the Groutz-Blaivas anti-incontinence surgery response score. The scores were statistically compared as binary categories at mean 111-day and 60.8-month followup. RESULTS: A total of 13 women underwent a simple sling incision for catheter dependent urinary retention after sling surgery, and 11 patients (mean age 73.4 years) were available for long-term followup (60.8 months). The simple sling incision procedure was completed an average of 65 days (range 36 to 235) after original sling placement. Mean post-void residual urine volume at least 1 month after sling surgery was 289 ml (range 75 to 500). At a mean followup of 60.8 months, no patient required catheterization. Of 11 patients 5 wore no pads. There was no statistical difference in leakage episodes per day (p = 1.0), pads per day (p = 0.3), or patient perceived condition (p = 0.3) during long-term followup. The mean Groutz-Blaivas score did not change statistically during the 5-year followup period (p = 0.6). CONCLUSIONS: Midline simple sling incision provides relief of catheter dependent obstruction following fascia lata sling surgery while preserving urinary continence in the majority of patients during a 5-year followup period. PMID- 16217329 TI - Total and immediate daytime and nighttime continence with a right colonic neobladder--What makes it possible? An 11-year followup. AB - PURPOSE: We report the long-term functional results of the colonic neobladder and provide the physiological and urodynamic foundations for them. MATERIALS AND METHODS: From March 1993 to February 2004, 38 patients with cystectomy received a neobladder constructed from detubularized, remodeled right colon and intact cecum following our design at the urology service at our institution. Most patients underwent urodynamics and videourodynamics as postoperative followup. We defined total continence as not using any protection whatsoever (neither pads nor a night alarm), which was immediate upon removing the urethral catheter. RESULTS: A total of 37 patients achieved total daytime continence immediately and the remaining 1 was totally continent after 30 days (100%). Nighttime continence was total and immediate in 36 patients (92%) and satisfactory in 2. Micturition was immediate, satisfactory and total in 37 patients with an average maximum flow of more than 26 ml per second. The patient who did not achieve micturition required clean intermittent catheterization. Videourodynamic studies revealed that continence resulted from the low pressure developed in the large capacity reservoir (more than 600 ml) and from intact intestine haustral contractions, which ejected urine toward the detubularized and remodeled area. In turn, micturition was attained through a combination of abdominal wall tension and mass contractions of the nondetubularized segment, which generated a pressure of more than 100 cm. CONCLUSIONS: The functional results of this new neobladder are significantly greater than those achieved with other procedures already described in the literature because of the different way in which it functions. PMID- 16217330 TI - The natural history of lower urinary tract dysfunction in men: minimum 10-year urodynamic followup of transurethral resection of prostate for bladder outlet obstruction. AB - PURPOSE: Despite long-term symptomatic and uroflowmetry studies following transurethral prostate resection (TURP) there are sparse pressure flow data. Consequently there is minimal information to account for the long-term symptomatic failure and flow rate decrease seen with time following early improvements after surgery. MATERIALS AND METHODS: Men older than 45 years who were investigated at our department between 1972 and 1986, diagnosed with bladder outlet obstruction and elected surgical intervention were invited for repeat symptomatic and urodynamic assessment. Identical methods were used, allowing direct comparison of results. RESULTS: A total of 1,068 men were initially diagnosed with bladder outlet obstruction, of whom 428 (40%) died in the interim. Of the men who were followed 217 underwent TURP with a mean followup since surgery of 13.0 years. A significant, sustained decrease in the majority of symptoms and improvements of urodynamic parameters was seen. Long-term symptomatic failure and decreased flow rate were principally associated with detrusor under activity (DUA) rather than obstruction. Presentation predictive factors for the future development of DUA were decreased detrusor contractility and a lesser degree of obstruction. CONCLUSIONS: This unique long-term study provides valuable information on surgically treated bladder outlet obstruction. The association of long-term failure following surgery with DUA emphasizes the importance of pressure flow studies before repeat surgery. However, our faith in the long-term efficacy of TURP is justified. PMID- 16217331 TI - Troponin T and N-terminal pro-brain natriuretic peptide changes in patients undergoing transurethral resection of the prostate. AB - PURPOSE: We investigated whether transurethral resection of the prostate (TURP) caused subclinical myocardial damage or cardiac dysfunction by measuring troponin T (Trop T) and N-terminal pro-brain natriuretic peptide (pro-BNP). MATERIALS AND METHODS: A total of 52 consenting patients took part in this study. All had a detailed medical history including cardiac history taken. On the day of the operation all patients had troponin T, pro-BNP, full blood count and urea, electrolytes and creatinine measured preoperatively. A preoperative and postoperative electrocardiogram was performed. Patients in renal failure were excluded from analysis. During the operations factors such as blood loss, operative time, tissue resected and fluid absorption were monitored. On postoperative day 1 all the previously mentioned tests were repeated. RESULTS: Mean patient age was 71 years (range 52 to 85). Eight patients had a history of associated cardiac problems. Mean preoperative and postoperative hemoglobin were 14.1 gm/dl (range 10.5 to 17) and 13.3 gm/dl (range 9.9 to 16.2), respectively. None of the patients had significant (greater than 1,000 ml) fluid absorption during TURP, which was calculated using ethanol tagged glycine. Mean blood loss measured with a photometer was 129.7 ml (range 0 to 1,800). Mean operative time was 28.4 minutes (range 5 to 50) and mean weight of prostatic tissue resected was 15.2 gm (range 1 to 47). Preoperative Trop T was less than 0.01 mcg/ml in all patients and mean pro-BNP was 39.2 pg/ml (range 0.5 to 866). Postoperative Trop T was less than 0.01 mcg/ml in all but 1 patient who experienced chest pain after TURP and had an increased Trop T (0.28 mcg/ml). Mean postoperative pro-BNP was 54.57 pg/ml (range 1 to 679). A total of 37 patients had an increase in pro-BNP which was still within the reference range for the age group. There were no significant electrocardiogram changes postoperatively. The Trop T changes were not statistically significant (Wilcoxon sign ranked test p = 0.31) although they may be clinically significant. CONCLUSIONS: Our study indicates that in patients with no prior cardiac history TURP does not cause myocardial damage indicated by nonincrease of Trop T. There are slight increases in pro-BNP after TURP in some patients although the exact clinical significance is uncertain. PMID- 16217332 TI - Development of a multiregional United States Spanish version of the international prostate symptom score and the benign prostatic hyperplasia impact index. AB - PURPOSE: The International Prostate Symptom Score (I-PSS) and Benign Prostatic Hyperplasia Impact Index (BII) have gained widespread use in clinical practice and clinical trials. Although Spanish translations of the I-PSS are available, to our knowledge none was developed for the Spanish speaking population in the United States using a methodology to ensure appropriateness for the diverse United States Spanish speaking population. An existing translation intended for another Spanish speaking country, such as Mexico, or a translation developed without input from each language group may not be understood by those who immigrated from other Latin American regions. Hence, the development of a Spanish translation for the United States should involve input from translators from each region of Latin America. MATERIALS AND METHODS: We reviewed and modified an existing United States Spanish translation of I-PSS using a multiregional reconciliation panel comprised of representatives from each of the major Spanish language groups in the United States. For BII full translation methodology was used to develop a translation for the United States, including 2 forward translations using translators from more than 1 region, a multiregional reconciliation panel meeting, a back translation evaluation, cognitive debriefing interviews with representatives from each language group, developer review, a final evaluation for consistency and proofreading. RESULTS: The revised I-PSS better reflects common Spanish wording in the United States, while the BII translation was confirmed to be comprehended by Spanish speakers in the United States originating from multiple regions of Latin America. CONCLUSIONS: United States Spanish translations of patient reported outcome measures should consider the diversity of the growing Spanish speaking population in the United States to ensure comprehension across the broad population originating from the multiple regions of Latin America. PMID- 16217333 TI - New theories in interstitial cystitis. PMID- 16217334 TI - Full publication of abstracts of randomised controlled trials published at International Continence Society meetings. PMID- 16217335 TI - Prostate botulinum A toxin injection--an alternative treatment for benign prostatic obstruction in poor surgical candidates. PMID- 16217336 TI - Effect of dutasteride on the detection of prostate cancer in men with benign prostatic hyperplasia. PMID- 16217337 TI - Three-dimensional ultrasonography in evaluation of benign prostatic hyperplasia. PMID- 16217338 TI - Patients with a large prostate show a higher prevalence of androgenetic alopecia. PMID- 16217339 TI - Terminal loop cutaneous ureterostomy in renal transplantation: an under utilized urinary diversion technique. AB - PURPOSE: We evaluated the effectiveness of terminal loop cutaneous ureterostomy as a means of urinary drainage in kidney transplant recipients during a 20-year period. MATERIALS AND METHODS: Five cadaveric and 2 living related patients underwent kidney transplantation with terminal loop cutaneous ureterostomy between 1984 and 2004. These patients had no usable bladder or they were not suitable candidates for intermittent catheterization. RESULTS: Followup was 20 months to 17 years. One patient underwent stomal revision 5 months after renal transplantation. Current serum creatinine 4 years later was 166 mumol/l. The remaining 6 patients had no evidence of ureteral obstruction and rarely had bacteriuria or urinary tract infections. Four patients had a functioning allograft with normal serum creatinine. One patient died with a normally functioning allograft and the remaining patient lost his graft due to chronic rejection. No patient in this series lost the graft due to a urological cause. Overall outcomes included excellent allograft function with minimal infection or stomal stenotic complications. CONCLUSIONS: Terminal loop cutaneous ureterostomy is a simple, safe and alternative means of urinary diversion in patients with renal transplant and a defunctionalized lower urinary tract. PMID- 16217341 TI - Acute kidney transplant failure following transurethral bladder polyp fulguration. PMID- 16217340 TI - Increase in utilization of percutaneous renal artery interventions by Medicare beneficiaries, 1996-2000. PMID- 16217342 TI - Stump transureteroureterostomy in en bloc kidney transplantation. PMID- 16217343 TI - Kidney paired donation and optimizing the use of live donor organs. PMID- 16217344 TI - Psychiatric morbidity is frequently undetected in patients with erectile dysfunction. AB - PURPOSE: We investigated the prevalence of psychiatric morbidity in patients with erectile dysfunction (ED). MATERIALS AND METHODS: Consecutive patients who attended an andrology outpatient clinic complaining of ED were recruited for study purposes. ED severity was evaluated by the International Index of Erectile Function erectile function domain score. The psychiatric diagnosis was established by a semistructured clinical interview as well as by the Mini International Neuropsychiatric Interview 5 Diagnostic and Statistical Manual of Mental Disorders-IV. The Beck Depression Inventory was also incorporated as a complementary tool to assess depressive symptom severity. RESULTS: The study included 103 patients 20 to 76 years old (mean age +/- SD 47.06 +/- 14.78) with ED. Most patients were classified with moderate and severe ED (26.2% and 44.7%, respectively). A detectable psychiatric condition was present in 63.1% of the patients, including depressive disorders in 25.2%, anxiety disorders in 11.7%, depression-anxiety comorbidity in 6.8% and personality disorders in 5.8%. A positive psychiatric anamnesis was identified in 41 cases, while 24 were newly diagnosed. No significant association was found between the duration and severity of ED, and psychiatric morbidity or the severity of depressive symptoms (p > 0.05). A positive association was found between the severity of depressive symptoms and the patient tendency to discuss the problem with their partner (p < 0.01). CONCLUSIONS: Psychiatric morbidity is highly prevalent in patients with ED, potentially affecting treatment outcome. Because lifetime psychological problems were reported by more than half of the patients, a psychosocial history seems mandatory. Partner support appeared to have a significant role in the patient psychological state. PMID- 16217346 TI - Sexual dysfunction in women with hyperprolactinemia: a pilot study report. AB - PURPOSE: Hyperprolactinemia is a common hormonal disorder in women that may affect the phases of female sexual function (FSD). We investigated sexual function in patients with hyperprolactinemia. MATERIAL AND METHODS: A total of 25 women with primary hyperprolactinemia and 16 age matched voluntary healthy women who served as the as control group were evaluated with a detailed medical and sexual history, including a female sexual function index (FSFI) questionnaire and the Beck Depression Inventory. Serum prolactin, dehydroepiandrosterone sulfate, free testosterone, androstenedione, 17alpha-hydroxyprogesterone, estradiol, free thyroxin and thyrotropin were measured. These variables were compared statistically between the 2 groups. RESULTS: Except for prolactin serum hormone levels in women with hyperprolactinemia were not different from those in the control group. The median total FSFI score was 23.40 (IQR 17.70 to 27.30) in the hyperprolactinemic group, whereas healthy women had a median total FSFI score of 31.10 (IQR 27.55 to 32.88, p < 0.0001). FSD was diagnosed in 22 of 25 patients (88%), while 4 of 16 healthy women (25%) had FSD (p = 0.03). Desire (p = 0.001), arousal (p < 0.0001), lubrication (p = 0.001), orgasm (p = 0.001), satisfaction (p = 0.07) and pain (p = 0.003) domain scores were also significantly lower in women with hyperprolactinemia. Total FSFI (p = 0.009, r = -0.405), desire (p = 0.001, r = -0.512), arousal (p = 0.002, r = -0.466), orgasm (p = 0.026, r = 0.348) and satisfaction (p = 0.041, r = -0.320) scores negatively correlated with mean prolactin but not with the other hormones measured. CONCLUSIONS: A significant percent of women with hyperprolactinemia whom we evaluated had sexual dysfunction. No hormonal changes other than prolactin and no depression was found as a cause of FSD. PMID- 16217345 TI - Corpus cavernosum electromyography during morning naps in healthy volunteers: further evidence that corpus cavernosum potentials reflect sympathetically mediated activity. AB - PURPOSE: We assessed the practicability of corpus cavernosum (CC) electromyography (EMG) in volunteers during morning naps in the laboratory and further validated this method. MATERIALS AND METHODS: A total of 11 healthy volunteers with a mean age of 23.8 years (range 19 to 31) were included. CC-EMG was started between 6:30 a.m. and 7 a.m. Two surface electrodes were placed at the base of the penis bilaterally and a reference electrode was placed on 1 kneecap. A strain gauge or Barlow gauge (Behavioral Technology, Salt Lake City, Utah) was used to monitor changes in penile circumference. Subjects were asked to sleep. Recording duration was 2 to 3 hours. Two recordings were performed per subject. RESULTS: Full erection was observed on 17 of the 22 recordings (77%), partial erection was noted on 3 (14%) and no tumescence was observed on the other 2 (9%). CC potentials consistently disappeared during tumescence and erection, while continuous CC potential oscillations reappeared during detumescence. During flaccidity bursts of CC potentials and electrical silence were recorded. Penile shrinkage was observed to accompany CC potentials but not to accompany electrical silence. CONCLUSIONS: CC-EMG during morning naps is a practical and valid method for investigating CC electrophysiology. CC-EMG signal patterns during tumescence, detumescence and flaccidity fit the existing theory that CC potentials reflect cavernous smooth muscle sympathetically mediated activity. PMID- 16217347 TI - Surgery or assisted reproduction? A decision analysis of treatment costs in male infertility. AB - PURPOSE: Assisted reproductive technology (ART), including in vitro fertilization and intracytoplasmic sperm injection, is routinely used to treat male factor infertility. Because of the success of ART, the optimal method to achieve pregnancy with male infertility is controversial. Two examples in which ART competes with traditional male infertility treatments are varicocelectomy and vasectomy reversal. We used formal decision analysis to estimate and compare the cost-effectiveness of surgical therapy and ART for varicocele and vasectomy reversal. MATERIALS AND METHODS: Decision analysis models were created for infertile men seeking paternity with varicocele and with post-vasectomy obstruction. Outcome probabilities applied to the model were derived from institutional and published sources. Costs of interventions were calculated from institutional data. Sensitivity analyses determined which elements were most important and, thus, were used to calculate threshold values. RESULTS: Vasectomy reversal is as cost-effective as ART if bilateral vasovasostomy can be performed. However, if unilateral or bilateral vasoepididymostomy is required, sperm retrieval/intracytoplasmic sperm injection may be more cost-effective due to lower patency rates. Vasectomy reversal is more cost-effective across all pregnancy rates provided that patency rates are greater than 79%. Surgical repair of varicocele is more cost-effective when the postoperative pregnancy rate is greater than 14% in men with a preoperative total motile sperm count of less than 10 million sperm and greater than 45% in men with greater than 10 million total motile sperm. CONCLUSIONS: A decision analysis based comparison of ART and classic surgical therapy suggests that varicocelectomy and vasectomy reversal are the most economical treatments in many cases of infertility due to these lesions. Tailoring the decision models to individual centers permits more accurate comparisons using specific costs as well as the surgical outcomes and results of ART. PMID- 16217348 TI - Erectile dysfunction and andropause symptoms in infertile men. AB - PURPOSE: We evaluated the prevalence of andropause symptoms and erectile dysfunction in our infertile population. MATERIALS AND METHODS: A total of 302 consecutive men presenting for infertility evaluation and 60 consecutive men with proven fertility seeking vasectomy (controls) were administered the Androgen Deficiency in the Aging Male and Sexual Health Inventory for Men (SHIM) questionnaires. Information regarding other clinical parameters, including seminal parameters, was collected by review of patient charts. RESULTS: Of the 302 infertile men screened, 38% reported significant andropause symptoms and 28% had abnormal SHIM scores. Of the subgroup of infertile men with nonobstructive azoospermia, 25% reported andropause symptoms and 27% had an abnormal SHIM score. In the fertile group 21% reported andropause symptoms and only 11% had an abnormal SHIM score. The prevalence of erectile dysfunction in infertile men was significantly higher than in the fertile controls (p = 0.007). CONCLUSIONS: Andropause symptoms and erectile dysfunction are common among infertile men, affecting approximately 38% of this population. This finding suggests that the population of infertile men should be carefully screened to identify and treat those with erectile dysfunction. PMID- 16217350 TI - Evaluation of the role of pudendal nerve integrity in female sexual function using noninvasive techniques. PMID- 16217349 TI - Pelviscrotal vasovasostomy: refining and troubleshooting. AB - PURPOSE: Obstruction of the vas deferens in the inguinal canal may occur as a sequel of inguinal surgery. The condition is occurs in 26.7% of cases following childhood herniotomy. Open surgery in the inguinal region for anastomosing the remnants of the vas is difficult and associated with high morbidity. We have previously reported an alternative based on anastomosing the pelvic vas deferens (harvested laparoscopically) to the scrotal vas deferens and bypassing the inguinal vas. This technique, termed Shaeer pelviscrotal vasovasostomy, is easier to perform with much less morbidity. In this study we present the results of performing the procedure bilaterally at the same session as well as technique troubleshooting. MATERIALS AND METHODS: A total of 25 patients with azoospermia due to inguinal obstruction of the vas deferens underwent unilateral (15) or bilateral (10) surgery. Patients were followed for 1 year. RESULTS: Of the 25 patients 17 (68%) had a sperm concentration of between 11.88 and 17 million per ml. Some patients who remained azoospermic underwent reoperation and the obstacles to a successful anastomosis were analyzed and resolved. CONCLUSIONS: Shaeer vasovasostomy is a practical approach to inguinal obstruction of the vas deferens. It enables a reliable anastomosis, simultaneous bilateral repair and lower morbidity in terms of wound healing and hernia as well as a shorter convalescence. PMID- 16217351 TI - Practice patterns of physician members of the American Urogynecologic Society regarding female sexual dysfunction: results of a national survey. PMID- 16217352 TI - Benign prostatic hyperplasia (BPH) and prostatitis: prevalence of painful ejaculation in men with clinical BPH. PMID- 16217353 TI - Sexual, psychological and dyadic qualities of the prostate cancer 'couple'. PMID- 16217354 TI - Alfuzosin 10 mg once daily improves sexual function in men with lower urinary tract symptoms and concomitant sexual dysfunction. PMID- 16217355 TI - Correlation between LUTS (AUA-SS) and erectile dysfunction (SHIM) in an age matched racially diverse male population: data from the prostate cancer awareness week (PCAW). PMID- 16217356 TI - Secondhand tobacco smoke impairs neurogenic and endothelium-dependent relaxation of rabbit corpus cavernosum smooth muscle: improvement with chronic oral administration of L-arginine. PMID- 16217357 TI - Psychobiological correlates of smoking in patients with erectile dysfunction. PMID- 16217358 TI - Inheritance of varicoceles. PMID- 16217359 TI - Induction of spermatogenesis in azoospermic men after internal spermatic vein embolization for the treatment of varicocele. PMID- 16217360 TI - The high incidence of meiotic errors increases with decreased sperm count in severe male factor infertilities. PMID- 16217361 TI - High frequency of Y chromosome microdeletions in idiopathic azoospermic men with high follicle-stimulating hormone levels. PMID- 16217362 TI - Beneficial effect of microsurgical varicocelectomy on human sperm DNA integrity. PMID- 16217363 TI - Health related quality of life after orthotopic neobladder construction and its comparison with normative values in the Japanese population. AB - PURPOSE: We report the difference in quality of life (QOL) between patients with an orthotopic neobladder and normative values in the age matched Japanese population. MATERIALS AND METHODS: Between November 1996 and June 2003, 75 patients underwent orthotopic neobladder construction at our institution, of whom 47 males and 9 females were enrolled in this study. A total of 19 patients were excluded because of death or insufficient followup (less than 6 months). Two types of questionnaire, namely the RAND 36-Item Health Survey, version 2 and the Functional Assessment of Cancer Therapy-Bladder Cancer, were mailed to all 56 patients. The 48 patients (86%) who returned the completed forms were included in our analysis. Postoperative QOL was compared with age matched normative values in the control Japanese population. RESULTS: Overall there was no significant difference in any scale between patients with a neobladder and the age matched control Japanese population. The patient bodily pain score in the seventh decade of life and the role-physical score in those older than 70 years were better than age matched normative values in the Japanese population (p < 0.005 and p < 0.05, respectively). Male patients in the seventh decade of life were more likely to have better QOL in the physical functioning, role-physical and role-emotional subscales in addition to the bodily pain subscale compared to age matched control values (p < 0.05, < 0.05, < 0.001 and < 0.001, respectively). Likewise in male patients older than 70 years role-physical scores remained better than age matched normative values in the Japanese population (p < 0.01). Further analysis of bladder cancer related QOL showed that patients followed more than 24 months were more likely to have a lower incontinence score than those followed less than 24 months (p < 0.05). In addition, the continence related QOL change alone did not affect any health related QOL scales, as analyzed by RAND 36-Intem Health Survey. CONCLUSIONS: Our study shows that there is no essential difference in health related QOL between patients with a neobladder and the age matched Japanese control population. The current results provide some primary evidence to help in decision making and eliminate potential anxiety about worse QOL after cystectomy. PMID- 16217364 TI - The costs of caring for erectile dysfunction in a managed care setting: evidence from a large national claims database. AB - PURPOSE: We examined the direct costs of erectile dysfunction (ED) empirically. MATERIALS AND METHODS: A naturalistic cohort study was done in 285,436 patients with ED and 51 health plans that covered 28 million lives nationwide from 1999 through 2001. Based on claims that had an ED related diagnosis code, procedure code or medication code we categorized the cost structure of ED care and calculated the annual costs of ED care per patient with ED, per user and per member monthly for individual and for all categories of ED care. RESULTS: A patient with ED in a health plan spent about an average of 83.91 dollars in 1999, 95.41 dollars in 2000 and 119.26 dollars in 2001 for ED care. In 2001, 37.08% of ED care costs per patient with ED were spent on phosphodiesterase type 5 (PDE-5) inhibitor therapy, 14.36% were spent on physician office visits, 10.19% were spent on diagnosis procedures, 8.45% were spent on testosterone hormone therapy, 3.85% were spent on penile prosthesis implantation, 4.41% were spent on intracavernous injection, 2.68% were spent on alprostadil pellet insertion and 0.81% was spent on vacuum erection devices. Of the 7 commonly used ED treatments PDE-5 inhibitor therapy has the lowest annual cost per user. CONCLUSIONS: In 2001 ED imposed a 122,669 dollars annual burden to a health plan with 100,000 members, that is or 0.108 dollars per member monthly. Each patient with ED spent 119.26 dollars annually for all ED related services or treatments. Of the 7 commonly used treatments PDE-5 inhibitor therapy had the lowest annual cost per user. PMID- 16217365 TI - What makes a medical student avoid or enter a career in urology? Results of an international survey. AB - PURPOSE: To recruit the best and brightest medical students, it is crucial to understand what causes a medical student to pursue or avoid a career in urology. We performed a survey of 2 cohorts of residency applicants to elucidate these determinants. MATERIALS AND METHODS: All 410 applicants to the 2003-2004 United States urology residency match and all 624 individuals who applied to a Boston area emergency medicine (EM) residency program were invited to participate in the study. The on-line survey for the urology applicants asked the research question "What caused you to pursue the specialty of urology?" The EM applicant survey asked the research question "What caused you NOT to pursue a career in a surgical subspecialty, such as urology?" Qualitative responses were analyzed for themes by 2 researchers and independently coded. RESULTS: Sixty percent (248 of 410) of urology applicants and 40% (252 of 624) of EM applicants completed the survey. Thematic coding of the qualitative responses yielded an inter-rater agreement of 89% to 94%. Positive determinants cited by urology applicants included the mix of medicine/surgery, the diversity of urological procedures, and clinical exposure to the field. Negative determinants cited by EM applicants included the narrowness of the specialty, an unattractive lifestyle, and the demands of a surgical residency. No significant correlations were noted between themes cited and participants' genders, degrees or medical school nationalities. CONCLUSIONS: Ensuring that medical students have clinical exposure to urology, receive appropriate mentorship, and develop realistic perceptions of the specialty may substantially facilitate recruitment. PMID- 16217366 TI - Prospective evaluation of remote, interactive videoconferencing to enhance urology resident education: the genitourinary teleteaching initiative. AB - PURPOSE: Changes in referral patterns and resource allocation into Centers of Excellence affect the educational experience of urology trainees by altering resident exposure to patients and clinicians, especially at sites where subspecialty deficiencies exist. Access to educators at Centers of Excellence using interactive videoconferencing technology may facilitate residency training objectives and enhance trainees' overall educational experience. We prospectively evaluated the implementation of this technology at tertiary care teaching centers to enhance urology resident education. MATERIALS AND METHODS: Using videoconferencing technology, urology residents at the University of Western Ontario (London, Canada) participated in a series of didactic, interactive pediatric urology teleteaching seminars. These were presented by an expert pediatric urologist from the Hospital for Sick Children, Toronto, Canada. Using a 5-point Likert scale (1-strongly disagree, 5-strongly agree), participants responded to statements pertaining to seminar content, technology and ease of use at the completion of each session. The results were subsequently tabulated and evaluated to determine the effectiveness and accessibility of the program in providing expert pediatric urological education to residents at a remote urology training program. RESULTS: The entire urology resident staff from postgraduate year 1 to 5 participated in the seminar program. The overall acceptance of this medium was high (mean score 4.5). The quality of presentation, as well as picture and sound quality, all received mean scores greater than 4. Participants indicated that their ability to interact with the presenter was not inhibited by using this medium. All participants agreed that they would use this technology in the future (mean score 4.5) and that the presentation would not be improved if the presenter were on-site. Due to preexisting technology at both centers, no direct cost was incurred throughout the study. CONCLUSIONS: Our experience suggests that interactive teleteaching using readily available, existing technology, is a cost-effective and accepted method of providing trainees with an appropriate educational experience. In centers where subspecialty deficiencies exist, this medium may provide residents with the necessary education requirements of their respective programs without the need for costly teacher (or student) travel. Continual improvements in technology as well as the addition of multiple sites will increased this medium's impact in the future. PMID- 16217367 TI - Report of the national survey of Urology Program Directors: attitudes and actions regarding the accreditation council for graduate medical education regulations. AB - PURPOSE: Residency programs in the United States are held accountable for achieving outcomes based on the new Accreditation Council for Graduate Medical Education (ACGME) regulations for improving resident fatigue and education. A national survey of Urology Program Directors was conducted to determine the attitudes toward the new regulations and the actions taken to implement them. MATERIALS AND METHODS: A national survey was developed by an ad hoc committee and distributed electronically. A total of 24 multiple-choice customized Likert scale questions were used to obtain opinions from Urology Program Directors across the country. The data were evaluated using statistical analysis. RESULTS: There was an 88% response rate to the survey. Of the respondents 53% demonstrated a statistically significant association between the size of the program and the degree to which the program was involved in addressing the duty hours standard (p = 0.015). A majority of program directors reported involvement in educating residents and faculty about the ACGME competencies. According to program directors, faculty members were negative about training residents in an 80-hour workweek and residents seemed to be apathetic. CONCLUSIONS: The national survey of Urology Program Directors has demonstrated that most programs have started to attend to the cultural change occurring in GME. Although there are some pockets of resistance to the ACGME regulations, the duty hours have been folded into most programs with little apprehension. Barriers to implementation of the Core Competencies discovered in this survey included inadequate validated instrumentation, inadequate staffing, and insufficient financial support. PMID- 16217368 TI - Clinical and physical determinants for toxicity of 125-I seed prostate brachytherapy. PMID- 16217369 TI - Facing erectile dysfunction due to prostate cancer treatment: perspectives of men and their partners. PMID- 16217370 TI - The changing nature of women's sexual health concerns through the midlife years. PMID- 16217371 TI - End stage renal disease in patients with Wilms tumor: results from the National Wilms Tumor Study Group and the United States Renal Data System. AB - PURPOSE: We sought to assess accurately the full spectrum of end stage renal disease (ESRD) in Wilms tumor survivors by combining the unique resources of the National Wilms Tumor Study Group (NWTSG) and the United States Renal Data System (USRDS), and to confirm preliminary reports of an increased incidence of ESRD in patients with the Wilms tumor-aniridia syndrome (WAGR). MATERIALS AND METHODS: ESRD was ascertained in 5,910 patients enrolled in NWTSG studies during 1969 to 1994 by record linkage to USRDS and by direct followup. Cumulative ESRD incidence was estimated accounting for intercurrent mortality. RESULTS: Of 115 cases of ESRD 10 (9%) were ascertained by the NWTSG alone, 13 (11%) by USRDS alone and 92 (80%) by both. Cumulative incidence of ESRD at 20 years from diagnosis of unilateral Wilms tumor was 74% for 17 patients with the Denys-Drash syndrome, 36% for 37 patients with WAGR, 7% for 125 male patients with hypospadias or cryptorchidism (genitourinary [GU] anomalies) and 0.6% for 5,347 patients with none of these conditions. The incidence of ESRD after diagnosis of bilateral Wilms tumor was 50% for the Denys-Drash syndrome (6 patients), 90% for WAGR (10), 25% for GU anomaly (25) and 12% for other (409). ESRD in patients with WAGR or GU anomalies tended to occur relatively late, often during or after adolescence. CONCLUSIONS: The risk of ESRD is remarkably low for the majority of patients with Wilms tumor. However, those with WAGR or associated GU anomalies are at higher risk and should be screened indefinitely to facilitate prospective treatment of impaired renal function. PMID- 16217372 TI - Urothelial neoplasms in patients 20 years or younger: a clinicopathological analysis using the world health organization 2004 bladder consensus classification. AB - PURPOSE: Urothelial neoplasms in patients younger than 20 years are rare, with conflicting data regarding clinical outcomes. MATERIALS AND METHODS: We identified 23 patients 4 to 20 years old with urothelial neoplasms, reclassified the microscopic diagnoses using the 2004 WHO/International Society of Urologic Pathology grading classification and collected data on presentation, risk factors and outcomes. RESULTS: Pathological grading revealed 2 urothelial papillomas, 10 papillary urothelial neoplasms of low malignant potential (PUNLMPs), and 8 low grade and 3 high grade papillary urothelial cancers, all without invasion. Mean patient age was 13.2 years (range 4 to 20), 19 patients were male and 19 presented with gross hematuria. All lesions were solitary and measured 0.1 to 6 cm. One patient had a history of smoking and 1 had parents who smoked. Three patients (13%) had recurrences classified as either urothelial papilloma (1) or PUNLMP (2). All patients were alive with no evidence of disease after a mean followup of 4.5 years (range 6 months to 13 years). CONCLUSIONS: Urothelial neoplasms in individuals younger than 20 years more commonly occur in males and are predominantly low grade with a favorable clinical outcome. Before the current classification system the 10 patients with a diagnosis of PUNLMP would have been classified as having papillary carcinoma. Thus, the diagnostic category of PUNLMP allowed 43.5% of patients in this series to avoid being labeled with "cancer" at a young age. PMID- 16217373 TI - Loss of interstitial cells of Cajal and gap junction protein connexin 43 at the vesicoureteral junction in children with vesicoureteral reflux. AB - PURPOSE: Symmetrical muscular contractions and unidirectional peristalsis are prerequisites for competent valve function at the ureterovesical junction. Interstitial cells of Cajal (ICCs) are pacemaker cells that create and coordinate peristaltic motility. We investigated ureteral endings immunohistochemically to elucidate the presence of c-kit positive ICCs as well as the occurrence of gap junction protein connexin 43 in children with vesicoureteral reflux (VUR) to identify a possible pathogenic factor of an insufficient antireflux mechanism. MATERIALS AND METHODS: Distal ureteral specimens were obtained from 27 children undergoing antireflux surgery. Routine histological paraffin embedded sections were immunostained detecting the c-kit proto-oncogene to study the presence of ICCs as well as connexin 43 positive cells to determine the gap junction density within the ureteral wall. Age matched nonrefluxing ureteral endings served as controls. All investigations were done using high power field magnification for semiquantitative analysis and statistically analyzed. RESULTS: ICCs were present in refluxing and nonrefluxing ureteral endings. Healthy individuals displayed significantly more ICCs than children with VUR. Connexin 43 immunoreactivity was significantly decreased in all refluxing ureteral specimens, whereas it was homogeneously distributed in normal controls. CONCLUSIONS: C-kit positive ICCs are found at the ureterovesical junction. Refluxing ureteral endings significantly lack these pacemaker cells, implying a malfunctioning valve mechanism permitting VUR. A substantial decrease in gap junctions in this region adversely affects intercellular signaling, aggravating coordinated peristalsis, which is essential for a competent anti-reflux mechanism. PMID- 16217375 TI - Bilateral extravesical ureteral reimplantation in toilet trained children: Is 1 day hospitalization without urinary retention possible? AB - PURPOSE: Bilateral extravesical ureteral reimplantation has been associated with urinary retention. We developed a critical pathway and modification of surgical technique to determine whether the bilateral extravesical procedure could be performed in toilet trained children with patients discharged home after a 1-day hospitalization and without urinary retention. MATERIALS AND METHODS: A total of 50 consecutive toilet trained children were evaluated after undergoing bilateral extravesical ureteral reimplantation using a modified technique that limits ureteral dissection, ureteral mobilization and detrusor dissection to as distally as possible so that a 5:1 ratio of tunnel length to ureteral diameter can be accomplished. No surgical dissection occurs in proximity to the obliterated umbilical artery, nor is the artery ligated. Patients follow a strict postoperative critical pathway, and parents receive extensive preoperative and postoperative education. A child is required to fulfill 5 strict criteria to be discharged from the hospital. RESULTS: Patient age ranged from 1.9 to 12.8 years (mean 4.9), with 37 girls and 13 boys participating. All patients were discharged home on postoperative day 1. All patients were able to void postoperatively without any instances of urinary retention. None of the children had acute urinary tract infections or required rehospitalization. All patients had radiographic resolution of the vesicoureteral reflux on postoperative voiding cystourethrogram. CONCLUSIONS: To our knowledge this is the first study to demonstrate that bilateral extravesical ureteroneocystostomy can be performed in selected patients without postoperative urinary retention and with uniform hospital discharge in 1 day. The critical pathway and limited dissection extravesical approach are essential for this success. PMID- 16217374 TI - Unilateral extravesical ureteral reimplantation in children performed on an outpatient basis. AB - PURPOSE: Unilateral extravesical ureteral reimplantation is comparable to intravesical procedures for resolution of primary vesicoureteral reflux (VUR). Defining whether this operation can be consistently performed on an outpatient basis is important. MATERIALS AND METHODS: A total of 80 patients with unilateral VUR were treated with extravesical ureteral reimplantation, of whom 20 were treated on an inpatient basis and 60 on an outpatient basis. We retrospectively reviewed these groups and conducted a telephone survey to evaluate overall patient satisfaction, and pain and nausea on postoperative days 1 and 14. RESULTS: There were no significant differences in age, gender, laterality or operative time between the groups. Average length of hospital stay was 31.25 hours (range 20 to 120) for the inpatient group and 6.6 hours (3.25 to 11.20) for the outpatient group. Average intravenous narcotic use in the inpatient group was 0.39 mg/kg, compared to 0.14 mg/kg for the outpatient group (p < 0.005), and included 1.76 mg/kg ketorolac in inpatients and 0.74 ketorolac in outpatients (p < 0.005), and 0.2 mg/kg ondansetron in inpatients and 0.12 mg/kg ondansetron in outpatients (p = 0.004). Four of the 60 outpatients (6.7%) were either hospitalized postoperatively or rehospitalized on postoperative day 1. The results of the survey for the 2 groups were not significantly different. CONCLUSIONS: Extravesical ureteral reimplantation for unilateral VUR may be performed without compromise in quality on an outpatient basis with significantly less use of intravenous analgesics and anti-emetics. PMID- 16217376 TI - The effect of biofeedback treatment on voiding and urodynamic parameters in children with voiding dysfunction. AB - PURPOSE: We studied the effects of biofeedback treatment on voiding and urodynamic parameters in children with voiding dysfunction. MATERIALS AND METHODS: We prospectively studied 188 children between June 1998 and November 2003. Biofeedback was performed with a urodynamics processor that enables simultaneous recording of urine flow and electromyography, and visual display of flow/electromyography activity. At the beginning of the study this treatment was performed once weekly. After patients understood the concept and performed homework regularly sessions were scheduled at 3 to 4-week intervals and continued for 6 months. All children were evaluated at 6 months and again at 2 years after completing biofeedback training. RESULTS: The number of the children completing biofeedback was 168 (89.4%). Improvement was obtained in all parameters, ranging from 59.2% to 87.8% at 6 months. Maximum improvement was acquired with flattened voiding in 65 of 74 children (87.8%), whereas the least improvement was acquired with daytime wetting in 58 of 98 children (59.2%). These improvements continued at 2-year followup, and ranged from 53.1% to 87.3%. While the rate of improvement for nocturnal enuresis, staccato voiding, detrusor-sphincter dyssynergia, vesicoureteral reflux and urinary tract infection had increased, the remaining parameters had decreased at 2-year followup. CONCLUSIONS: Biofeedback training is a simple, effective and well tolerated treatment modality in children for various parameters resulting from bladder dysfunction. Motivation and willingness to participate in biofeedback treatment are important selection criteria. PMID- 16217377 TI - Congenital mid ureteral strictures. AB - PURPOSE: Congenital mid ureteral stricture is rare. We report 7 cases, and discuss the differences in preoperative evaluation and surgical management compared to other obstructive entities. MATERIALS AND METHODS: Medical records and imaging studies of 7 children identified with mid ureteral strictures between 1998 and 2002 were reviewed retrospectively. Five newborns presented with prenatal hydronephrosis, and 2 children presented at age 15 years, one in the course of evaluation of blunt trauma, and one due to pain and abdominal mass. Imaging studies included renal ultrasound, voiding cystourethrography, radionuclide renography and computerized tomography. All patients underwent retrograde pyelography. Pathological examination of each specimen was undertaken at the respective institutions. RESULTS: Prenatal hydronephrosis was the most common presentation. There were no urinary tract infections. All patients had significant obstruction on the affected side. No patient had vesicoureteral reflux. After imaging but before surgery the urinary obstruction was believed to be at the ureteropelvic junction in 4 patients and the ureterovesical junction in 2, and secondary to posterior urethral valves in 1. At cystoscopy all of the affected ureters had a normally located and normally configured orifice. Retrograde pyelography led to an accurate diagnosis of mid ureteral narrowing in all patients. Six patients underwent ureteroureterostomy, all of whom had satisfactory outcomes. In 1 of these patients contralateral nephrectomy was performed due to nonfunction of the multicystic dysplastic kidney. The remaining patient underwent nephrectomy for ipsilateral end stage kidney disease and hydronephrosis. In this patient the ureters were stenotic and suggested asymmetry in the thickness of the muscular coat, perhaps secondary to extrinsic compression. CONCLUSIONS: Congenital mid ureteral stricture is rare. Renal ultrasound and radionuclide renography alone do not reliably demonstrate the site of obstruction. Retrograde pyelography at the time of surgical correction of presumed ureteral obstruction is an important adjunct for correctly identifying the site of narrowing in the affected ureteral segment, unless the ureter has been imaged with another modality. PMID- 16217378 TI - Adolescent varicocele repair: long-term results and comparison of surgical techniques according to optical magnification use in 100 cases at a single university hospital. AB - PURPOSE: We review the long-term results of varicocele repair, and compare the complication rates of varicocelectomy techniques according to optical magnification use in adolescents with varicocele at a single university hospital. MATERIALS AND METHODS: We prospectively studied 100 males 7 to 19 years old with clinical palpable varicocele. Of the patients 52 (52%) underwent left unilateral varicocelectomy and 48 (48%) underwent bilateral varicocelectomy. Varicocelectomy was performed using a microscope in 49 patients (79 sites), loupe magnification in 25 (35 sites) and no magnification in 26 (34 sites) using either a subinguinal or inguinal approach. Postoperative complications were compared in all patients based on technique. Preoperative and postoperative serum hormone values and semen parameters were compared in 33 patients. RESULTS: Mean postoperative followup was 30.4 +/- 13.06 months (12 to 65). Total motile sperm count increased from 22.6 million +/- 5.16 million to 64.53 million +/- 12.3 million postoperatively, which was statistically significant (p = 0.002). Postoperative recurrence rates were 0% in cases managed by microsurgical varicocelectomy, 2.9% in those where loupe magnification was used and 8.8% in those where no magnification was used. Postoperative hydrocele rates in these cases were 0%, 2.9% and 5.9%, respectively. The highest rates of recurrence and hydrocele were observed in cases where no magnification was used, compared to those managed by microsurgery (p = 0.03 and p = 0.116, respectively). CONCLUSIONS: Adolescent varicocele repair improves semen parameters and is a safe method with low recurrence and low complication rates. Our study suggests that the postoperative complication rate significantly decreases with use of higher magnification, such as microscopy. Microsurgical varicocele repair is the best technique with the lowest postoperative rates of recurrence and hydrocele in the treatment of adolescent varicoceles. PMID- 16217379 TI - Testicular microlithiasis and antisperm antibodies following testicular biopsy in boys with cryptorchidism. AB - PURPOSE: Testicular biopsy (TBx) performed during orchiopexy in boys with cryptorchidism can help to predict future semen analyses and possibly identify patients at risk for testicular cancer. It has been theorized that TBx can be detrimental to the long-term health of the testis. We examined testicular microlithiasis (TM) and antisperm antibody (ASA) production in cryptorchid TBx. MATERIALS AND METHODS: A total of 112 males underwent fertility evaluation. These patients had previously undergone orchiopexy and bilateral TBx (mean age 8.6 years) for unilateral or bilateral undescended testis. At a mean age of 19.6 years all patients underwent physical examination and scrotal ultrasound for evaluation of testis size, echotexture and abnormalities. Of 112 patients 57 also underwent direct Immunobead(R) assay with positive controls. Moving sperm free and/or bound to IgG bead complex were counted in the preparation. RESULTS: A total of 29 patients had bilateral and 83 had unilateral undescended testis. Of the 112 patients 26 were black and 86 were white. Four black patients (15.4%) and 4 white patients (4.7%) had testicular microlithiasis. Three patients had bilateral and 5 had unilateral diffuse TM. No tunica albuginea scars or testis masses were noted. In the 57 patients who underwent direct Immunobead assay no semen sample demonstrated evidence of forming bead-sperm complex, ie none had direct ASA. CONCLUSIONS: No patient exhibited evidence of direct ASA. Prepubertal open TBx does not increase the rate of TM. We found no evidence of additive testicular damage associated with TBx at the time of orchiopexy. PMID- 16217380 TI - Gender assignment in female congenital adrenal hyperplasia: a difficult experience. PMID- 16217381 TI - Culture clash involving intersex. PMID- 16217382 TI - Urinary levels of interleukin-6 and interleukin-8 in children with urinary tract infections to age 2. PMID- 16217384 TI - Myceliatomas (Aspergillus niger) in the renal collecting system and ureters. PMID- 16217383 TI - "Burnt-out" testicular germ cell tumors. PMID- 16217385 TI - Photodynamic therapy with PAD-S31, a new hydrophilic chlorin photosensitizer, in an orthotopic rat bladder tumor model. AB - PURPOSE: PAD-S31 (13,17-bis (1-carboxypropion) carbamoylethyl-3-ethenyl-8 ethoxyiminoethylidene-7-hydroxy-2,7,12,18-tetramethyl-porphyrin sodium) (Photochemical Co., Ltd., Okayama, Japan), 1 of the latest second-generation photosensitizers, has hydrophilic characteristics and excitation wavelengths of around 670 nm. Using an orthotopic rat bladder tumor model we investigated the biodistribution of PAD-S31 and assessed the antitumor effects of photodynamic therapy (PDT) with PAD-S31. MATERIALS AND METHODS: An orthotopic rat bladder tumor was established by implanting AY-27 cells in the bladder wall. After intravenous PAD-S31 administration the accumulation of PAD-S31 in the tumor and normal bladder wall was investigated by a fluorometric technique. One or 3 hours after intravenous administration of PAD-S31 (5 mg/kg) bladder tumors in rats were transurethrally irradiated at 100 mW/cm with a light dose of 50 to 200 J/cm. The efficacy of PDT was evaluated 7 days later by observation with an ultrathin cystoscope and histopathological examination. RESULTS: The ratio of PAD-S31 concentration in tumor tissue to that in normal bladder wall was more than 1 at all time points and it achieved a maximum (more than 10) 150 to 240 minutes after PAD-S31 administration. All rats that were irradiated at 100 J/cm 3 hours after PAD-S31 administration showed more than 50% tumor destruction. When the light dose was more than 150 J/cm, more than half of the rats showed complete tumor eradication, of which the average size was 6 mm. CONCLUSIONS: We report that PDT using PAD-S31 is effective for destroying bladder tumors in an orthotopic rat model. These experimental results suggest that this therapy could be a clinically promising method for the treatment of patients with bladder cancer. PMID- 16217386 TI - Modulation of mitogen-activated protein kinase cascades by differentiation-1 protein: acquired drug resistance of hormone independent prostate cancer cells. AB - PURPOSE: The inhibitor of differentiation-1 protein (Id-1) is over expressed in multidrug resistance prostate cancer cells. We determined the effect of Id-1 expression and its underlying pathways on the development of multidrug resistance in prostate cancer. MATERIALS AND METHODS: AT3 cells were transfected with the Id 1 gene or a blank vector. Id-1 mRNA expression was determined by reverse transcriptase-polymerase chain reaction and Id-1 protein content was detected by immunoblot and flow cytometry. Cellular cytotoxicity was determined by MTT (microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay (Sigma Chemical Co., St. Louis, Missouri). The activation and expression of mitogen-activated protein kinase (MAPK) were measured by transactivation assay and Western blotting, respectively. RESULTS: Id-1 overproduction drove AT3 cells to become resistant to chemotherapeutic agents but did not induce mdr-1 gene expression. The p38MAPK and c-jun N-terminal kinase (JNK) pathways were suppressed, which correlated with increased Id-1 expression. No significant change in extracellular signal-regulated kinase (ERK) activation was observed in Id-1 transfectants compared with that of AT3 or vector control. Treatment of Id-1 expressing cells with p38MAPK and JNK inhibitors resulted in decreased doxorubicin induced apoptosis. In contrast, Id-1 expressing cells treated with ERK inhibitor made cells more sensitive to drug induced apoptosis. CONCLUSIONS: Up-regulation of Id-1 was found in prostate cancer multidrug resistant cells. Sustained ERK activation, and JNK and p38MAPK inhibition by Id-1 in cells may confer drug resistance. These changes in MAPKs could be a mechanism for the acquisition of multidrug resistance in prostate cancer. PMID- 16217387 TI - Urinary response to an oxalic acid load is influenced by the timing of calcium loading in rats. AB - PURPOSE: Dietary intake of calcium or dairy products has been shown to decrease urinary oxalate excretion by limiting its intestinal absorption. However, not enough attention has been given to whether there is any benefit from altering the schedule of ingesting calcium and oxalate. Therefore, we investigated the effects of changes in the timing of calcium and oxalate loading on urinary oxalate excretion. MATERIALS AND METHODS: Male Wistar rats weighing 180 to 200 gm were fasted and randomized into several groups. They were then administered normal saline or oxalic acid with or without calcium or milk. Calcium or milk was given immediately, or 5, 10, 15 or 30 minutes before or after the oxalate load. All treatments were given via gastrostomy. Urine samples were collected by bladder puncture just before administration and at hourly intervals up to 5 hours afterward. Urinary oxalate was measured by capillary electrophoresis. RESULTS: Urinary oxalate increased after the administration of oxalate alone, while it decreased when oxalate was combined with calcium or milk. Urinary oxalate showed a smaller increment when calcium or milk was given before than after oxalate loading, and it was much smaller when calcium or milk was given immediately before oxalate. CONCLUSIONS: Prior calcium loading appears to have a positive influence on decreasing oxalic acid absorption from the intestinal tract. Therefore, calcium or dairy products should always be ingested before a meal rich in oxalate to prevent oxalate absorption and decrease urinary oxalate excretion. PMID- 16217388 TI - Effects of tolterodine on an overactive bladder depend on suppression of C-fiber bladder afferent activity in rats. AB - PURPOSE: We determined whether the effects of antimuscarinics depend on the suppression of C-fiber bladder afferent nerves. We administered tolterodine intravenously or intravesically. MATERIALS AND METHODS: To induce C-fiber bladder afferent nerve desensitization resiniferatoxin (RTX) (0.3 mg/kg) was injected subcutaneously in female Sprague-Dawley rats 2 days prior to left middle cerebral artery occlusion (MCAO). As controls, we used rats treated with ethanol and saline vehicle (VEH). Insertion of a polyethylene catheter through the bladder dome and MCAO were performed using halothane anesthesia. The effects of intravenous (0.2 to 2000 nM/kg) or intravesical (0.2 or 2 nM) tolterodine, an antimuscarinic agent, on cystometrography were investigated in conscious rats with a cerebral infarct (CI). Tolterodine was instilled intravesically for 30 minutes and cystometry was repeated. RESULTS: Bladder capacity (BC) was markedly decreased after MCAO in RTX treated (RTX-CI) and VEH treated (VEH-CI) rats. Low tolterodine doses (0.2 or 2 nM/kg) significantly increased BC in VEH-CI rats without increasing residual volume but it had no effects on BC in RTX-CI rats. At the highest dose (2,000 nM/kg) the drug significantly decreased bladder contraction pressure and increased residual volume in RTX-CI and VEH-CI rats. Intravesical administration of tolterodine (0.2 or 2 nM) significantly increased BC in VEH-CI rats. However, tolterodine had no effect on BC in RTX-CI rats. CONCLUSIONS: These results suggest that at low doses tolterodine exerts an inhibitory effect on C-fiber bladder afferent nerves, thereby, improving BC during the storage phase. PMID- 16217389 TI - Tissue reactions of 5 sling materials and tissue material detachment strength of 4 synthetic mesh materials in a rabbit model. AB - PURPOSE: We evaluated tissue reactions to 5 sling materials used in tension-free vaginal tape (TVT), intravaginal slingplasty (IVS), polypropylene mesh hernia repair, the suprapubic approach to suburethral polypropylene tape (SPARC) and cadaveric fascia lata procedures. We also compared the mesh-to-tissue attachment strength of 4 sling mesh materials (TVT, IVS, surgical polypropylene mesh and SPARC) at on days 2, 7, 15 and 30 after implantation. MATERIALS AND METHODS: A total of 20 female New Zealand White rabbits were randomized to group 1-2 days, group 2-7 days, group 3-15 days and group 4-30 days. After the rabbits were anesthetized an 8 cm midline incision was made for rectus muscle access, and 0.5 x 1 cm pieces of TVT, IVS, SPARC, surgical polypropylene mesh and cadaveric fascia lata were sewn to the rectus muscle with direct contact. At the same time 4 subfascial tunnels in the medial surface of the upper extremities were prepared, and 1.0 x 0.5 cm strips of TVT, IVS, SPARC and polypropylene mesh were implanted in each tunnel. On days 2, 7, 15 and 30 after implantation mechanical testing was performed to define tissue detachment strength. The strips of 5 sling materials were then harvested with the surrounding tissue. Specimens were studied by light microscopy. RESULTS: Mean detachment strength, that is the minimum weight needed to move the mesh, of the synthetic meshes from days 2 to 30 were 291.6 to 2,390.0 gm for TVT, 178.4 to 2,160.0 gm for SPARC, 188.4 to 1,850.0 gm for hernia mesh and 92.8 to 1,510.0 gm for IVS (at all data points TVT vs IVS p < 0.05). Light microscopy revealed a quite uniform tissue reaction with a sign of marked acute inflammation in and around the mesh fibers on days 2 and 7 after implantation. All meshes showed stable fibrosis and muscle infiltration on day 30. CONCLUSIONS: All 5 synthetic sling materials produce similar tissue reactions beginning soon after implantation. Cadaveric fascia lata persisted in tissue with remarkable perifascial fibrosis at day 30. When comparing the 4 polypropylene mesh materials; the attachment capacity of TVT was superior and that of IVS was the least of the 4. TVT was statistically better than IVS at all data points. SPARC and hernia mesh provided results similar to those of TVT. PMID- 16217390 TI - Processed lipoaspirate cells for tissue engineering of the lower urinary tract: implications for the treatment of stress urinary incontinence and bladder reconstruction. AB - PURPOSE: We performed a pilot study to investigate the ability of human adipose derived, multipotent stem cells to be delivered to and survive within bladder and urethral smooth muscle. MATERIALS AND METHODS: Lipoaspirate was acquired from female patients undergoing liposuction. The lipoaspirate was processed to yield a pluripotent population of processed lipoaspirate (PLA) cells. For tissue delivery PLA cells were fluorescent labeled and suspended in Hanks' balanced salt solution (Sigma Chemical Co., St. Louis, Missouri). To assess PLA viability in multiple animal models 8 Rnu athymic rats (Charles River, Wilmington, Massachusetts) and 6 SCID mice (Taconic Farms, Oxnard, California) underwent laparotomy and injection of PLA cells into the bladder and urethra. An additional 8 rats underwent sham injection of Hanks' balanced salt solution alone. Experimental and control animals were sacrificed 2, 4, 8 and 12 weeks after injection, and the bladders and urethras were analyzed. RESULTS: Self-regenerating, pluripotent PLA cells were easily isolated from human adipose tissue. Evaluation 2, 4, 8 and 12 weeks after injection demonstrated PLA cell viability and incorporation into the recipient smooth muscle. Eight weeks following injection PLA cells demonstrated in vivo expression of alpha-smooth muscle actin, an early marker of smooth muscle differentiation. CONCLUSIONS: PLA cells are an easily accessible source of pluripotent cells, making them ideal for tissue regeneration. PLA cells remain viable up to 12 weeks in the lower urinary tract. Human PLA cells injected into the urinary tract show morphological and phenotypic evidence of smooth muscle incorporation and differentiation with time. PLA cells may provide a feasible and cost-effective cell source for urinary tract reconstruction. PMID- 16217391 TI - Expression of survivin and apoptotic biomarkers in benign prostatic hyperplasia. AB - PURPOSE: We assessed the differential expression of survivin and other apoptotic markers in stromal and epithelial compartments of benign prostatic hyperplasia (BPH) and normal prostate tissue. MATERIALS AND METHODS: Immunohistochemical staining for survivin, and transforming growth factor-beta1 and its receptors was done in normal prostate and BPH areas from 114 consecutive patients who underwent radical prostatectomy. Moreover, staining for survivin, Bcl-2, Bax, caspase-3 and Ki-67 was performed in prostate specimens from 23 consecutive patients who underwent open prostatectomy and 4 young patients who underwent cystoprostatectomy and had a normal prostate. RESULTS: Survivin and Bcl-2 expression increased incrementally from normal prostate to epithelial BPH to stromal BPH. Caspase-3 expression was higher in BPH epithelium than in BPH stroma, which in turn was higher than that in normal prostate. Ki-67 was significantly over expressed in BPH stroma and epithelium. Survivin expression in BPH tissue correlated with International Prostate Symptom Score, quality of life, post-void residual urine volume, maximum urine flow rate and transforming growth factor-beta1 expression. CONCLUSIONS: Survivin is over expressed in BPH and it correlates with BPH parameters. Increases in proliferation and inhibition of apoptosis have a role in BPH. PMID- 16217392 TI - Immunolocalization of estrogen receptor alpha and beta in human fetal prostate. AB - PURPOSE: We examined the immunolocalization of estrogen receptor (ER)alpha and ERbeta in the human fetal prostate. MATERIALS AND METHODS: Tissue sections from human fetal prostates at 7 to 22 weeks of gestation were stained with antibodies to ERalpha, ERbeta, and cytokeratin 10 and 14. RESULTS: ERalpha expression was not detected until 15 weeks of gestation with sparse staining in the utricle. By 19 weeks increased ERalpha expression was seen in the luminal cells of the ventral urogenital epithelium (UGE), basal cells of the dorsal UGE, utricle, distal periurethral ducts, peripheral stroma and posterior prostatic duct. K14 was detected in basal cells of the UGE and in several posterior acini. At 22 weeks ERalpha expression was more intense in all of these areas. ERbeta was expressed throughout the UGE, ejaculatory ducts, mullerian ducts and entire stroma at 7 weeks. Intense ERbeta staining was observed in these areas and in the prostatic buds by 8 weeks with persistent intense staining through 22 weeks. CONCLUSIONS: To our knowledge we report the first immunolocalization of ERalpha in the human fetal prostate and the earliest demonstration of ERbeta expression in the prostate at 7 weeks of gestation. ERbeta expression is intense during ductal morphogenesis, suggesting a role in normal glandular growth and proliferation. The induction of squamous metaplasia in the UGE, distal periurethral ducts and utricle is associated with ERalpha expression in these areas, while the induction of squamous metaplasia in peripheral prostatic acini is associated with peripheral stromal ERalpha expression. This study suggests estrogen signaling pathways in the human fetal prostate via ERalpha that involve epithelial-epithelial and epithelial-stromal interactions. PMID- 16217393 TI - Poly(Adenosine diphosphate-ribose) polymerase inhibition preserves erectile function in rats after cavernous nerve injury. AB - PURPOSE: We investigated the involvement of poly(adenosine diphosphate-ribose) (PAR) polymerase (PARP) activation in the development of erectile dysfunction and the therapeutic benefit of the potent PARP inhibitor INO-1001 (Inotek Pharmaceuticals Corp., Beverly, Massachusetts) in a bilateral cavernous nerve crush injury (BCNCI) model in rats. MATERIALS AND METHODS: Sprague-Dawley rats were divided into 3 groups, namely sham treated, BCNCI plus vehicle and BCNCI plus the PARP inhibitor INO-1001. One week after surgical intervention all groups underwent in vivo cavernous nerve stimulation. PAR activation, nitrotyrosine and inducible nitric oxide synthase were evaluated by immunohistochemistry and serum levels of INO-1001 were measured by high performance liquid chromatography. Penile tissues were analyzed for levels of malondialdehyde and myeloperoxidase. Data sets were statistically compared in all groups. RESULTS: Neurogenic mediated erectile responses were evaluated. Mean intracavernous pressure (ICP), the ICP-to blood-pressure ratio and total ICP were significantly decreased in BCNCI plus vehicle rats. These values were not statistically different between the sham and PARP inhibitor treated groups. There was a marked decrease in PAR staining in the treatment group. There was a substantial increase in malondialdehyde tissue levels but not myeloperoxidase in response to BCNCI, which was unchanged with PARP inhibitor treatment. There was a marked increase in tyrosine nitration in the treatment group. Up-regulation of nitric oxide synthase and increased tyrosine nitration were not observed in the penile tissues of the treatment group. CONCLUSIONS: These data demonstrate that BCNCI in a rat model causes increased PARP activation, resulting in severe erectile dysfunction. Treatment with the PARP inhibitor INO-1001 decreases the degree of nitrosative stress, prevents PARP activation and provides significant cavernous neuroprotection, which in turn preserves erectile function. PMID- 16217394 TI - Erythropoietin promotes the recovery of erectile function following cavernous nerve injury. AB - PURPOSE: We investigated the effects of recombinant human (rh) erythropoietin (EPO) on erectile function recovery in a rat model of cavernous nerve (CN) injury. MATERIALS AND METHODS: Male rats underwent unilateral CN transection and excision of a 5 mm segment of the contralateral CN. One group received rhEPO (5,000 U/kg) subcutaneously daily for 14 days, while another received rhEPO 1 day and 1 hour prior to nerve injury. An additional group of animals was pretreated with 1 dose of darbepoetin (25 microg/kg). At 14 days following CN injury rats underwent erection physiology studies. Axonal regeneration was evaluated by electron microscopy. EPO receptor expression in the penis and major pelvic ganglion was evaluated immunohistochemically and by real-time polymerase chain reaction. RESULTS: Daily rhEPO effectively recovered erections after CN injury compared with saline treatment. Maximal intracavernous pressure area under the curve normalized to mean arterial pressure was significantly greater in EPO treated vs saline treated animals (p < 0.05). rhEPO and darbepoetin pretreatment was as effective as continuous 14-day therapy. EPO receptor expression was localized to neuronal cell bodies of the major pelvic ganglion, penile nerves and endothelial cells in the penis. Electron microscopy revealed significant improvement in axonal regeneration in rhEPO treated animals 14 days following injury compared to controls. CONCLUSIONS: EPO receptors are expressed in local neuronal and vascular tissues. Exogenous administration of rhEPO or darbepoetin in the setting of CN injury promotes erectile function recovery. This occurs through axonal regeneration of the injured nerve and possible penile protection. PMID- 16217395 TI - Urine markers for bladder cancer surveillance: a systematic review. PMID- 16217396 TI - Assessment by M-FISH of karyotypic complexity and cytogenetic evolution in bladder cancer in vitro. PMID- 16217397 TI - Effects of N-acetyl transferase 1 and 2 polymorphisms on bladder cancer risk in Caucasians. PMID- 16217398 TI - CDC91L1 (PIG-U) mRNA expression in urothelial cell carcinomas. PMID- 16217399 TI - Re: conservative management of small testicular tumors relative to carcinoma in situ prevalence. PMID- 16217400 TI - Re: renal cell carcinoma with renal vein and inferior vena caval involvement: clinicopathological features, surgical techniques and outcomes. PMID- 16217402 TI - Re: luteinizing hormone-releasing hormone agonist effects on skeletal muscle: how hormonal therapy in prostate cancer affects muscular strength. PMID- 16217404 TI - Re: individualization of the biopsy protocol according to the prostate gland volume for prostate cancer detection. PMID- 16217406 TI - Re: stone and pelvic urine culture and sensitivity are better than bladder urine as predictors of urosepsis following percutaneous nephrolithotomy: a prospective clinical study. PMID- 16217407 TI - Re: percutaneous nephrolithotomy for caliceal diverticular calculi: a novel single stage approach. PMID- 16217409 TI - Re: metabolic consequences and long-term complications of enterocystoplasty in children: a review. PMID- 16217411 TI - Re: the increasing incidence of newborn circumcision: data from the nationwide inpatient sample. PMID- 16217414 TI - When disaster strikes. AB - In response to the healthcare issues resulting from Hurricane Katrina, The Nurse Practitioner has compiled information from the Centers for Disease Control and Prevention (CDC) to educate nurse practitioners (NPs) about previously uncommon illnesses presenting in the wake of the disaster, CDC recommendations on immunizations and infection prevention, and guidelines for providing proper wound care. Visit http://www.cdc.gov for full text of these articles. PMID- 16217415 TI - NPs called to duty in Katrina aftermath. PMID- 16217416 TI - Damage caps--another view. PMID- 16217418 TI - Hypertension: catching the silent killer. PMID- 16217420 TI - Abnormal uterine bleeding. PMID- 16217426 TI - Unmasking the truth behind endocrine disruptors. AB - The increase in reproductive cancers and developmental problems over the past 70 years has led researchers to suspect environmental influences as a root cause. Evidence from wildlife and laboratory studies suggests that exposure to endocrine disruptors (EnDs) may be the cause. An EnD is a foreign substance or mixture that alters the function of the endocrine system. They can be found in food, water, soil, or air. Research into their possible role provides an opportunity to decrease modifiable risk factors. PMID- 16217427 TI - Field guide to pain part 2. Developing a plan of care. AB - In this second installment of the Field Guide to Pain, the authors describe pharmacological treatments, physical therapy, interventional treatments, and behavior therapy as viable options to incorporate into a plan of care. By using simple assessment tools and becoming familiar with the various classes of pain medication, primary care providers can facilitate treatment of patients in pain. PMID- 16217428 TI - Guide to care for patients. Obesity. PMID- 16217434 TI - Setting the stage for changing health behavior. PMID- 16217441 TI - Autogenous bone grafts and bone substitutes--tools and techniques: I. A 20,000 case experience in maxillofacial and craniofacial surgery. AB - Alloplastic bone substitutes can be used to alter facial contour. In contrast, autogenous bone grafts have a successful 80-year history of restoring facial contour as well as the basic functional support of the craniofacial skeleton. The traditional procedures for harvesting and using autogenous bone grafts are not obsolete. During the past 30 years, the techniques have been refined and new sources have been found, such as calvarial grafts. New tools were required and have been designed to make harvesting of grafts easier and faster for the surgeon and safer and less expensive for the patient. Four short articles under the heading of "Techniques and Tools" are presented addressing the harvesting of (1) iliac, (2) costal, (3) tibial, and (4) calvarial grafts. These articles are based on the experience of six surgeons using the same technique and instruments in more than 20,000 autogenous bone grafting procedures. (These figures represent the group experience as of 2001. Since then, one of the junior coauthors has retired, but the remaining five continue to harvest autogenous bone grafts on a regular basis. So, the group experience as of 2004 is in the range of 23,000 procedures). The-senior surgeon's experience of 9500 procedures spans a period of 50 years (from 1946 to 1996). For the other surgeons (10,500 procedures combined), the collection period was 25 years (from 1975 to 2000). PMID- 16217442 TI - Taking bone grafts from the anterior and posterior ilium--tools and techniques: II. A 6800-case experience in maxillofacial and craniofacial surgery. PMID- 16217443 TI - Taking long rib grafts for facial reconstruction--tools and techniques: III. A 2900-case experience in maxillofacial and craniofacial surgery. PMID- 16217444 TI - Taking tibial grafts in the diaphysis and upper epiphysis--tools and techniques: IV. A 650-case experience in maxillofacial and craniofacial surgery. PMID- 16217445 TI - Taking calvarial grafts, either split in situ or splitting of the parietal bone flap ex vivo--tools and techniques: V. A 9650-case experience in craniofacial and maxillofacial surgery. PMID- 16217446 TI - Complications of harvesting autogenous bone grafts: a group experience of 20,000 cases. PMID- 16217447 TI - Taking calvarial grafts--tools and techniques: VI. The splitting of a parietal bone "flap". PMID- 16217448 TI - On the maintenance and sharpening of instruments. PMID- 16217450 TI - Pre-expansion of mucosa-lined flap for lower eyelid reconstruction. PMID- 16217452 TI - Temporary orbital apex syndrome after repair of orbital wall fracture. PMID- 16217453 TI - The reversed Goldstein technique to correct a hypoplastic Cupid's bow following bilateral cleft lip repair. PMID- 16217455 TI - Parry-Romberg syndrome. PMID- 16217456 TI - Posttraumatic thumb reconstruction. AB - LEARNING OBJECTIVES: After reading this article, the reader should be able to: 1. Discuss the critical anatomic features of the thumb as they affect on reconstructive decision making. 2. Define the goals of reconstruction. 3. Discuss an algorithm for thumb reconstruction according to the level of amputation. 4. Understand the role of prosthetics in thumb reconstruction. BACKGROUND: The function of the thumb is critical to overall hand function. Uniquely endowed with anatomic features that allow circumduction and opposition, the thumb enables activities of pinch, grasp, and fine manipulation that are essential in daily life. Destruction of the thumb secondary to trauma represents a much more significant loss than would result from loss of any other digit. Therefore, significant effort has been focused on thumb reconstruction. Numerous techniques have been described, ranging from simple osteoplastic techniques to complex microsurgical procedures. With an appreciation of the unique anatomic properties of the thumb, the hand surgeon is better able to understand the goals of thumb reconstruction and to develop an algorithm for thumb reconstruction. With such an understanding, an individualized reconstructive plan can be developed for each patient. METHODS: A great many options are available for posttraumatic thumb reconstruction. Optimal results are obtained by pursuing an organized and logical approach to reconstruction based upon the level of tissue loss. Reconstruction methods depend on the location of the amputation and range from homodigital and heterodigital flaps to partial-toe transfer or a great-toe wrap-around flap to first-web-space deepening using Z-plasties, a dorsal rotation flap, or a distant flap, to distraction osteogenesis, lengthening of the thumb ray, spare parts from another injured digit in the acute setting for pollicization or heterotopic replantation, and microvascular toe transfer. RESULTS: Amputations in the distal third of the thumb are generally well-tolerated. The primary reconstructive issues are the restoration of a padded and sensate soft-tissue cover, as well as aesthetic considerations. First-web-space deepening will generally provide excellent results for amputations at the distal half of the middle third. In the proximal half of the middle third, lengthening of the thumb ray is generally required. Distraction lengthening of the first metacarpal is a useful and reliable technique that provides up to 3 cm of length without requiring complex microsurgical methods. Spare parts from another injured digit may be used in the acute setting for pollicization or heterotopic replantation. Microvascular toe transfer is an excellent option for elective reconstruction. However, other options also are available and may be more appropriate in some cases. Less ideal options include the various types of osteoplastic reconstruction. CONCLUSIONS: The reconstruction of posttraumatic thumb defects is a challenging and rewarding surgical endeavor. The value of a functioning thumb is immense, and its reconstruction is worthy of considerable effort. Despite the elegant reconstructive options available, the best results are obtained with replantation or revascularization whenever possible. Finally, the treatment plan always must be derived from a careful assessment of each patient's posttraumatic function and specific reconstructive needs. PMID- 16217459 TI - Carcinoma ex benign pleomorphic adenoma of the parotid gland. AB - BACKGROUND: Carcinoma in pleomorphic salivary adenoma is a common histologic subtype of primary parotid malignancy. METHODS: In this study, 28 patients (predominantly male) with histologically diagnosed carcinoma in pleomorphic salivary adenoma presenting over 10 years were retrospectively reviewed. RESULTS: Only 25 percent of patients had a previously treated pleomorphic salivary adenoma. Although the presenting features suggested malignancy in some cases, overall they were nonspecific, overlapping with the presentation of benign disease. Preoperative investigations included fine needle aspiration cytology, which was only 29-percent sensitive, and computed tomography and/or magnetic resonance imaging. There were 14 superficial and 12 total or radical parotidectomies. The facial nerve was resected en bloc with the tumor in nine cases and immediately reconstructed with good reanimation results in patients with recent-onset facial paresis. Only 44 percent of patients had a complete histologic tumor clearance, and this was the most significant determinant of survival (p < 0.01, log-rank analysis). The locoregional control rate was 66 percent at 5 years, but recurrent disease proved invariably fatal. Five-year disease-specific survival was 44 percent with a high rate of disease-specific mortality (87 percent). CONCLUSIONS: Carcinoma in pleomorphic salivary adenoma is very difficult to diagnose preoperatively. Fine needle aspiration cytology had a disappointingly low sensitivity for this tumor, potentially misdirecting surgical management. While good locoregional disease control could be achieved with surgery and radiotherapy, carcinoma in pleomorphic salivary adenoma was shown to be aggressive with a high disease-specific rate of mortality. Given that incomplete tumor resection was the most important prognostic factor, a high index of clinical suspicion, radical ablative surgery, and immediate soft-tissue and nerve reconstruction for proven cases are advocated. PMID- 16217457 TI - Salvage of silicone-treated facial deformities using autogenous free tissue transfer. AB - BACKGROUND: Silicone, in the form of injectable liquid and prosthetic blocks, has been used to correct facial contour defects. However, silicone also has been associated with many complications including malposition, dyschromia, dysesthesia, contractures, infection, and fistula. Complications related to injectable-liquid and prosthetic-block silicone use in treatment of facial deformities pose a challenging surgical dilemma. METHODS: Over a 15-year period, microvascular free flap technique has been used successfully as a surgical tool to correct severe soft-tissue complications of the face resulting from silicone treatment of facial contour deformities. Surgical treatment guidelines, methods, and results for treating this difficult problem with free tissue transfer are presented. RESULTS: In 23 patients, 24 free tissue transfers (22 inframammary extended circumflex scapular free flaps and two superficial inferior epigastric free flaps) were performed. Flap survival was 100 percent. Both hematoma and skin necrosis were encountered in three of 24 cases (12.5 percent). Revisional surgery combined with aesthetic surgical procedures was required in 20 of 24 cases (83 percent). CONCLUSIONS: We recommend the following surgical treatment guidelines for this unique subset of patients: silicone excision with immediate microvascular free flap reconstructive procedure when the skin and soft tissue are mildly to moderately affected, or silicone excision with delayed reconstructive procedure when the skin and soft tissue are severely affected. Free tissue transfer is a useful surgical tool for salvaging severe facial skin and soft-tissue deformities caused by previous silicone treatment. PMID- 16217460 TI - Expansion of the socket and orbit for congenital clinical anophthalmia. AB - BACKGROUND: Congenital clinical anophthalmia requires fitting an artificial eye, achieving a pleasing aspect of the palpebral apparatus, and stimulating growth of the orbit, as well as of the entire midface. METHODS: Out of 25 patients with clinical anophthalmia, 22 were treated according to a new protocol (first described by Wiese et al. in 1999) and followed for up to 6 years. In 10 patients, anophthalmia was bilateral, in nine patients, it was on the right side only and in three it was on the left side only. Treatment consisted of implantation of an osmotic expander to first stimulate growth of the conjunctival sac and eyelids followed by serial implantation of osmotic expanders to stimulate growth of the orbit (and thus of the midface). The material used was a copolymer of methylmethacrylate and N-vinylpyrrolidone. RESULTS: Fitting of an artificial eye was without any problem within the first year of life for the most part. Growth of the orbit was stimulated by more than 65 percent of a healthy one. Aesthetically, the results were very largely pleasing. Complications were encountered, especially when the wrong expander sizes had been implanted. CONCLUSION: Using osmotic expanders of this type, through a staged therapy of minor operations, promises to achieve all goals before the child enters school. PMID- 16217461 TI - Selective marginal mandibular neurectomy for treatment of the marginal mandibular lip deformity in patients with chronic unilateral facial palsies. AB - BACKGROUND: For unilateral facial palsies, reanimation techniques such as cross facial nerve grafting with free muscle transfer have been very successful at improving symmetrical lip elevation when smiling. However, these procedures do not address the residual asymmetry with respect to lip depression resulting from an uncorrected marginal mandibular nerve palsy. Techniques that do address this residual lip asymmetry have had variable results. Selective neurectomy of the unaffected marginal mandibular nerve has largely been abandoned as a treatment since it has been reported to result in severe oral functional and cosmetic deficiencies. The authors argue that selective marginal mandibular neurectomy is a reliable technique for treatment of the marginal mandibular lip deformity that does not compromise oral functionality. METHODS: Thirteen patients with unilateral facial palsies underwent selective marginal mandibular neurectomy of the unaffected side; the procedure was performed by the principal investigator. These patients were then evaluated on the basis of symmetry with smiling and oral functionality. Twelve of the patients were contacted and questioned with a standardized questionnaire. Twelve of the patients were evaluated independently for symmetry with smiling using randomized preoperative and postoperative photographs. RESULTS: Eleven of the 12 patients reported either improvement or no change in oral competence. Ten of the 12 patients reported either improvement or great improvement in symmetry with smiling. Improvement in symmetry with smiling was independently observed 77.2 percent of the time. CONCLUSION: These results support selective marginal mandibular neurectomy of the unaffected side in patients with unilateral facial palsies as a means of improving symmetry with smiling without compromising functionality. PMID- 16217462 TI - Prospective analysis of the outcome of surgical management of drooling in the pediatric population: a 10-year experience. AB - BACKGROUND: Sialorrhea is a common problem in the neurologically impaired pediatric patient and surgery is the mainstay of treatment for severely affected patients refractory to conservative or medical treatment. METHODS: The results of the surgical protocol used since 1993 at the Royal Children's Hospital in Melbourne, Australia, are reported. Seventy-two patients underwent bilateral submandibular duct transposition combined with bilateral sublingual gland excision. Patients were analyzed prospectively and reviewed at 1, 6, and 12 months, then at 2 and 5 years postoperatively. RESULTS: Of 67 patients with 2 years of complete follow-up, the median score for the frequency of drooling fell from 4.0 to 2.9 (p < 0.001) and the median severity-of-drooling score decreased from 4.8 to 3.0 (p < 0.001). There was a fall in the number of clothing/bib changes from a median of four per day to zero (p < 0.0001). Of 41 patients followed to 5 years, both the median frequency and severity of drooling scores remained at 3. Overall the majority of patients benefited from surgery. Thirteen patients (18 percent) experienced complications which were major in 9 percent. CONCLUSION: We have found bilateral sublingual gland excision in combination with bilateral submandibular duct transposition to be superior to bilateral submandibular duct transposition plus parotid duct ligation. The surgical treatment of sialorrhea continues to be a rewarding experience for our multidisciplinary team. PMID- 16217463 TI - Mulliken method of bilateral cleft lip repair: anthropometric evaluation. AB - BACKGROUND: Simultaneous surgical correction of bilateral cleft lip and nasal deformity is becoming more common. This is a major change from the conventional strategy of secondary nasal correction. METHODS: Thirty patients with bilateral cleft lip and nasal deformity were repaired using the Mulliken method between July of 1997 and December of 2002. This series was composed of 10 infants with bilateral complete cleft lip, 12 with bilateral incomplete cleft lip, and eight with bilateral asymmetric cleft lip, defined as complete on one side and incomplete on the other. A preoperative orthopedic appliance was employed in five infants who had severe collapse of the lateral maxillary segments. Mean age at the time of repair was 3.8 months. RESULT: Postoperative results were evaluated anthropometrically. The mean follow-up period was 3.8 years (range, 1.2 to 5.5 years). Nasal tip protrusion was low; interalar dimension was wide; columellar length was slightly short; upper cutaneous labial height was short; and vermilion mucosal height was near normal. All anthropometric values approached controls by 5 years. CONCLUSION: Our modified Mulliken method is effective in correction of all forms of bilateral cleft lip and nasal deformity. Further evaluation is needed for possible differences in outcome based on race and age at the time of operation. PMID- 16217465 TI - Treatment outcome in unilateral cleft lip and palate evaluated with the GOSLON yardstick: a meta-analysis of 1236 patients. AB - BACKGROUND: The goal of this study was to assess determinants for treatment outcome in unilateral cleft lip and palate, evaluated according to the Great Ormond Street London and Oslo (GOSLON) Yardstick and 5-year-index ratings by means of a meta-analysis. METHODS: Multiple databases were searched for publications in which patient groups were evaluated by GOSLON ranking or the GOSLON-like 5-year index. From the 15 selected publications, the following background variables could be extracted and were evaluated as determinants for treatment outcome in unilateral cleft lip and palate: year of birth, average age of the patient at the time of GOSLON classification, racial background, presence of Simonart's band, infant orthopedics, palatal closure before the age of 3 versus palatal closure at a later age, bone graft, and number of surgeons. RESULTS: The total number of patients included in the meta-analysis was 1236. Patients whose soft and hard palate were closed before the age of 3 presented significantly poorer (p = 0.003) GOSLON scores (mean score, 2.9; SD 0.4) than patients whose palate was closed at a later age (mean GOSLON score, 2.3; SD 0.2). CONCLUSIONS: Delayed palatal closure generally results in better dental arch relationships than early palatal closure. Well-designed, randomized clinical trials are required for further investigation of the optimal timing for palatal closure. PMID- 16217468 TI - The use of sentinel node biopsy in breast cancer patients undergoing skin sparing mastectomy and immediate autologous reconstruction. AB - BACKGROUND: Intraoperative frozen section examination of the sentinel node in breast cancer patients is associated with a high number of incorrect negative results with the sentinel node becoming positive in the permanent examination and necessitating a secondary axillary lymph node dissection. A reoperation of the axilla following skin-sparing mastectomy and immediate autologous tissue reconstruction may compromise the vascular pedicle of the flap and should be avoided. METHODS: Eighty breast cancer patients underwent skin-sparing mastectomy with immediate autologous reconstruction and sentinel node biopsy followed by axillary lymph node dissection irrespective of the result of the frozen section of the sentinel node. The goal of the study was to identify a subgroup of patients with incorrect negative sentinel node(s) in the frozen section who may forego a secondary axillary lymph node dissection due to a low risk of positive nonsentinel nodes. RESULTS: Frozen section examination of the sentinel node was negative in 58 patients and positive in 22 patients. Permanent histologic examination revealed tumor in 13 of 58 (22.4 percent) sentinel node(s) found negative in the frozen section. None of these 13 patients showed positive nodes in the axillary specimen, whereas nine of 22 patients with their metastases in the sentinel node found through intraoperative frozen section examination had additional positive nonsentinel node(s) (p = 0.001). CONCLUSIONS: Patients with incorrect negative sentinel node(s) found in the frozen section examination had a significantly decreased risk for additional positive nonsentinel node(s) compared with patients with sentinel node metastases found in the frozen section. However, to avoid a secondary axillary lymph node dissection, the authors suggest performing sentinel node biopsy before mastectomy under local anesthesia to have the permanent result of the sentinel node available before a planned reconstruction. PMID- 16217466 TI - Pelvic, abdominal, and chest wall reconstruction with AlloDerm in patients at increased risk for mesh-related complications. AB - BACKGROUND: The use of polypropylene mesh in the reconstruction of trunk defects increases complication rates when the mesh is placed directly over viscera or the operative site has been irradiated or contaminated with bacteria. An alternative is AlloDerm (decellularized human cadaveric dermis), which becomes vascularized and remodeled into autologous tissue after implantation. When used for fascial reconstruction, AlloDerm forms a strong repair, causes minimal abdominal adhesions, and resists infection. METHODS: We did a retrospective study of cancer patients at increased risk for mesh-related complications who underwent trunk reconstruction with AlloDerm over a 1-year period. Risk factors included unavoidable placement of mesh directly over the bowel or lung, perioperative irradiation, and/or bacterial contamination of the defect. The indications, defect characteristics, reconstructive techniques, complications, and surgical outcomes were evaluated. RESULTS: Thirteen patients were included in the study. Indications for reconstruction were oncologic resection, resection of enterocutaneous fistula, and/or ventral hernia repair. Seven patients had bacterial contamination at the operative site and seven patients received perioperative radiation. The mean musculofascial defect size was 435 cm. AlloDerm was placed directly over the bowel or lung in all patients. Nine patients required flap reconstruction, including 14 pedicled and two free flaps. The mean follow-up was 6.4 months. Complications occurred in six patients, however, there were no clinically evident mesh infections, hernias, or bulges. CONCLUSIONS: AlloDerm successfully can be used in reconstructions for large, complex pelvic, chest, and abdominal wall defects even when placed directly over viscera and when the operative field is irradiated and/or contaminated with bacteria. PMID- 16217469 TI - Prediction of postoperative seroma after latissimus dorsi breast reconstruction. AB - BACKGROUND: The latissimus dorsi flap has become a first-line option in reconstruction of the breast cancer patient. Donor-site seroma is a commonly described postoperative complication of the latissimus dorsi flap. METHODS: A retrospective chart review from 1998 to 2003 of all patients undergoing latissimus dorsi breast reconstruction was performed (n = 50). Age of the patients, timing of breast reconstruction, type of nodal dissection (axillary versus sentinel versus none), and chemotherapy status of the patients were examined. RESULTS: The overall incidence of seroma formation was 47 percent. Those patients who had undergone prior or concurrent nodal dissection at the time of breast reconstruction were found to have a higher incidence of seroma formation than patients who had no nodal dissection (52 percent versus 25 percent) (p = 0.15). Age also was a risk factor for seroma formation, as 63 percent of patients older than 50 had formed seroma as compared with 39 percent of those younger than age 50 (p = 0.08). CONCLUSION: The authors conclude that advanced age and the presence of nodal disruption before or concurrent with latissimus dorsi breast reconstruction are predictors of donor-site seroma formation. PMID- 16217471 TI - Fat injection to correct contour deformities in the reconstructed breast. AB - BACKGROUND: A ten-year, single-surgeon study of 37 patients from 1993 to 2003 who underwent fat injections to improve contour deformities in their reconstructed breasts was reviewed. METHODS: Fat was harvested from elsewhere in the body using a low-pressure syringe lipoaspiration system, washed gently with saline, and injected into depressions along the margins of reconstructed breasts. Blinded physician observers judged preoperative and postoperative photographs of breasts injected with fat and categorized the degree of contour improvement as substantial, minimal to moderate, or none. Complications of fat injections were noted. A total of 43 breasts in 37 patients were injected with autologous fat during 47 discrete events; some patients had the procedure repeated and some were treated bilaterally. Of the 43 treated breasts, 25 (58 percent) were reconstructed with implants, 17 (40 percent) were reconstructed with a TRAM (transverse rectus abdominis muscle) flap, and one (2 percent) was reconstructed with a TRAM and an implant. RESULTS: There were four complications (8.5 percent) in 47 treated breasts: one breast with cellulitis that resolved with antibiotics and three breasts with small, superficial lumps--two of which were biopsied and found to be liponecrotic cysts. Patient follow-up averaged 49 weeks, ranging from 3 weeks to 6 years. There was a substantial contour improvement in ten breasts (21 percent), minimal to moderate improvement in 30 breasts (64 percent), and no improvement in 7 breasts (15 percent). CONCLUSIONS: Although fat injection in and around the reconstructed breast has limitations, such as fat necrosis and need for repeated injections, our experience indicates that overall it is a very safe technique that can improve or correct significant contour deformities that otherwise would require more complicated, riskier procedures to improve. PMID- 16217470 TI - Standardized assessment of breast cancer surgical scars integrating the Vancouver Scar Scale, Short-Form McGill Pain Questionnaire, and patients' perspectives. AB - BACKGROUND: Currently, there is no standardized, comprehensive method to assess surgical scars after breast cancer surgery. This article evaluates the application of the Vancouver Scar Scale, in conjunction with patients' scar self rating and scar-related pain, in a cohort of breast cancer patients. METHODS: Data were prospectively collected in 59 women with breast cancer. Scar assessment comprised: 1. objective rating by pairs of independent observers using the Vancouver Scar Scale; 2. patient's ratings of the scar's physical parameters and overall satisfaction; and 3. pain assessment using the Short-Form McGill Pain Questionnaire. A total of 212 scar scores (59 pairs of breast/chest wall and 47 pairs of axillary scar scores) were generated by 13 observers: three physicians, five radiation therapists, and five nurses. Internal consistency was tested using Cronbach's alpha statistics. Interobserver reliability was evaluated with Spearman's rho and intraclass correlation coefficient computations. Convergent validity of the observer and patient ratings was examined with Spearman's correlation statistics. Linear regression analysis was performed to identify significant factors associated with Vancouver Scar Scale scores and patient satisfaction. RESULTS: The Vancouver Scar Scale, patient self-rating scale, and Short-Form McGill Pain Questionnaire had acceptable internal consistency (Cronbach's alpha 0.79, 0.64, and 0.72 respectively). Interobserver reliability using the Vancouver Scar Scale was significant with Spearman's correlation coefficients of 0.53 for pliability, 0.47 for scar height, 0.49 for vascularity, 0.54 for pigmentation, and 0.66 for overall score (all p values < 0.001). Significant agreement between observer and patient ratings of scar pliability (p = 0.01) and color (p = 0.001) was demonstrated. Mild to moderate pain was reported by more than 40 percent of patients. Patient satisfaction was significantly associated with self-rating of scar pliability and pain, but not Vancouver Scar Scale scores. CONCLUSIONS: The Vancouver Scar Scale is a reliable and valid tool to objectively evaluate scars after breast cancer surgery. Evaluation of scar-related pain and patients' scar rating and satisfaction provide additional information relevant to scar assessment. This integrated approach is feasible in a busy clinical setting to advance care and research in scar management for breast cancer patients. PMID- 16217472 TI - Psychosexual function in women who have undergone reduction mammaplasty. AB - BACKGROUND: Reduction mammaplasty substantially alleviates physical and psychological symptoms of mammary hyperplasia, but the effect on psychosexual function has not been analyzed so far. The aim of this study was to assess the impact of this common procedure on both operative and psychosexual satisfaction. METHODS: All women who underwent reduction mammaplasty (n = 90) at the Royal Free and University College London Hospitals between 1997 and 2000 were sent a series of questionnaires on their psychosexual health and satisfaction with operative outcome and postoperative sexual function. RESULTS: Of the 80 women who responded (response rate, 89 percent), 76 percent were satisfied with their operative results. Postoperative sexual satisfaction was improved in 28 percent, worsened in 19 percent, and unchanged in 53 percent of patients. Nobody dissatisfied with surgical outcome reported improved sexual satisfaction. A strong correlation was found between these two main outcome measures and General Health and Female Sexual Function Index scores. Significant correlations were also noted for breast shape, sensation, and symmetry (p < 0.001) and change in nipple sensation (p < 0.01). In contrast, breast size and scars did not significantly affect patient operative and sexual satisfaction (p > 0.1). When compared with the satisfied group, women unhappy with surgical results were more likely to suffer from postoperative anxiety (58 versus 18 percent) and depression (42 versus 8 percent). CONCLUSIONS: This study confirms that operative outcome following breast reduction does correlate with postoperative psychosexual function, mental health, and some breast parameters. Now that the link is identified, a prospective analysis may clarify the precise relationship between the patient's preoperative psychological status and operative and psychosexual satisfaction. PMID- 16217473 TI - Functional outcome for children with thumb aplasia undergoing pollicization. AB - BACKGROUND: Hand function is difficult to evaluate in young patients. It is helpful to assess young children after surgery for trauma or congenital anomaly to see how they cope as they progress through their developmental milestones. METHODS: Functional outcome in 10 children (12 upper extremities) who had previous pollicization for a congenitally absent or severely hypoplastic thumb were evaluated by standard radiographs, thumb total active range of motion, grip and pinch strength, parent questionnaire, modified Jebsen functional testing, and a pegboard Functional Dexterity Test. RESULTS: Grip strength was significantly less (p = 0.008) in the hands that had been operated on (mean, 2 kg) compared with the hands that had not been operated on (mean, 5.6 kg). Pinch strength was also significantly less (p = 0.008) in the hands that had been operated on (mean, 1.0 kg) compared with those that had not been operated on (mean, 2.1 kg). In most, hands that had been operated on and those that had not been operated on tested outside the 2-SD range of age-matched normals for pinch and grip strength and also for the Functional Dexterity Test. In contrast, total Jebsen Hand Function Test time was not significantly different from hands that had not been operated on, except that some subtests were significantly different, such as checker stacking (p = 0.016; mean difference, 7.2 seconds) and page turning (p = 0.031; mean difference, -10 seconds). The total active range of motion in hands that had been operated on and those that had not been operated on was also not significantly different. All children used their reconstructed thumbs in a normal pattern. Parent questionnaires revealed satisfaction with appearance and good social interactions. Functional activities showed greatest difficulty handling small objects, especially when simultaneous pinch strength was required, such as fastening buttons and small snaps. CONCLUSIONS: This study demonstrates the importance of evaluating multiple aspects of functional outcome for congenital hand problems and of using comparative age-appropriate validated norms. Pollicization is a rewarding procedure for children with thumb aplasia. PMID- 16217475 TI - The Suzuki frame for complex intraarticular fractures of the thumb. AB - BACKGROUND: Suzuki et al. in 1994 described the pins and rubber traction system for comminuted intraarticular fractures and fracture-dislocations of the proximal interphalangeal joint of the hand. The authors used the pins and rubber traction system to treat five complex fractures of the thumb. METHODS: From February of 1999 to April of 2003, five patients, four men and one woman, had a pins and rubber traction system applied for complex fractures of the interphalangeal joint (two patients) and metacarpophalangeal joint (three patients) of the thumb. The mean age of the patients was 35.4 years (range, 28 to 50 years). The system was applied for 4 weeks in all cases. RESULTS: The mean follow-up period was 23.6 months. The mean range of total movement for the metacarpophalangeal joint was 55 degrees (45 degrees of flexion and 10 degrees of extension). The two patients with the interphalangeal joint fractures achieved flexion of 0 to 65 degrees and 0 to 55 degrees, respectively. Joint space narrowing was observed in three cases. One patient had persistent swelling of the metacarpophalangeal joint 14 months after the initial treatment but had no functional deficit or pain. No pin track infection was observed in this series. CONCLUSIONS: Previous reports regarding the application of the pins and rubber traction system have focused on pilon fractures and dorsal fracture-dislocation of the proximal interphalangeal joint. In the present study, the authors described their experience with application of the Suzuki frame for complex intraarticular fractures of the thumb. The authors believe that the pins and rubber traction system can be used to treat difficult intraarticular fractures of the thumb, with very good functional results. PMID- 16217476 TI - Radial midpalmar island flap. AB - BACKGROUND: Although defects and contractures of the first web space and the thumb base can be a difficult problem, several local flaps have been used to reconstruct them. However, as most of these flaps are raised from the dorsal aspect of the thumb and the index finger, they may be unsuitable for reconstruction when defects or contractures of the first web space and the thumb base involve the dorsal skin. METHODS: In an attempt to resolve this problem, an island flap harvested from the radial aspect of the midpalm and based on the terminal branch of the superficial palmar arch was developed at Chonnam National University Medical School. From 1998 to 2003, this flap was used in 15 patients. The sizes of these flaps ranged from 2 to 3 cm in width and from 2.5 to 5 cm in length. RESULTS: All the flaps survived completely. The donor site was covered with a skin graft in six patients and closed primarily in nine. Healing of most donor sites was uncomplicated. However, in one patient, the skin graft on the donor site did not take completely as a result of subgraft hematomas and a subsequent split-thickness skin graft was required to cover the defect. CONCLUSIONS: Although there may be some variations of the vascular pedicle, the radial midpalmar island flap is reliable using careful dissection. This new flap may offer an alternative for reconstructing the first web space and the thumb base in selected patients. PMID- 16217477 TI - Pelvic ring reconstruction with the double-barreled vascularized fibular free flap. AB - BACKGROUND: Although hemipelvectomy has been the standard treatment for malignant tumors of the pelvis, limb salvage surgery is now the treatment of choice, even for patients with advanced tumors. For these patients, pelvic reconstruction is needed to maintain the stability of the pelvis and the spinal column and to allow ambulation. In this report, the authors' experiences with pelvic ring reconstruction are described. METHODS: Pelvic ring reconstruction with free double-barreled vascularized fibular grafts was performed after resection of malignant pelvic tumors in five patients. The graft was fixed with a fixation plate and screws in three patients and with the Cotrel-Dubousset rod system in two patients. After surgery, perioperative and postoperative findings were evaluated. RESULTS: In one patient, a pedicled rectus abdominis musculocutaneous flap was transferred to repair defects of the skin and underlying soft tissue. The free fibular graft was transferred successfully in four of five patients; however, the graft was removed in one patient because of infection with methicillin-resistant Staphylococcus aureus. After surgery, three of the four patients with successful grafts could walk with full weight bearing and without a cane; the fourth patient died as a result of multiple metastases to the lung before walking was attempted. CONCLUSIONS: The double-barreled fibular graft is well vascularized and can achieve satisfactory bone union. It is a safe and effective method for reconstructing the pelvic ring. Furthermore, the Cotrel Dubousset rod system can provide rigid fixation soon after surgery and is useful for early rehabilitation of walking. PMID- 16217478 TI - Tensor fasciae latae perforator flap: minimizing donor-site morbidity in the treatment of trochanteric pressure sores. AB - BACKGROUND: To report a new technique with less morbidity for coverage of trochanteric defects, an anatomical and clinical study was performed. METHODS: Twenty-four fresh cadavers were dissected. The following parameters were measured: origin, location, number, and length of the perforating vessels. In addition, a clinical study was performed on 21 patients with trochanteric pressure sores. RESULTS: The anatomical study of 24 fresh cadavers revealed the constant presence of perforator pedicles anterior to the greater trochanter, which provides an adequate arc of rotation arc for flap harvest without sacrificing the underlying muscles. The mean length of the pedicles was 9.59 +/- 2.16 cm. This flap is nourished by perforator vessels arising from the ascending branch of the lateral circumflex femoral artery, which arises from the deep femoral artery and runs through the intermuscular septum, tensor fasciae latae, and rectus femoralis muscles. In this study, flaps were raised based on perforators located preoperatively using a unidirectional Doppler probe. Good results were obtained with primary closure of the donor site, with only two donor site dehiscences. CONCLUSIONS: This flap is an alternative to myocutaneous flaps, as it preserves local musculature without functional sequelae in patients who walk. It also preserves the local musculature in the event of recurrence, as is usually seen in paralytic patients with pressure sores. PMID- 16217480 TI - Intrauterine fetal constraint induces chondrocyte apoptosis and premature ossification of the cranial base. AB - BACKGROUND: The spheno-occipital synchondrosis is an important growth center of the craniofacial skeleton and a primary site of malformation in syndromic forms of craniosynostosis. Clinical and laboratory investigations have demonstrated that premature closure of cranial vault sutures in nonsyndromic craniosynostosis is associated with characteristic alterations in cranial base morphology. However, a causal link between premature fusion of calvarial sutures and changes in the cranial base remains elusive. The purpose of these experiments was to test the hypothesis that intrauterine head constraint produces ultrastructural changes in the spheno-occipital synchondroses of fetal mice. METHODS: Fetal constraint was induced through uterine cerclage of six pregnant C57Bl/6 mice on the eighteenth day of gestation. Fetuses were harvested after growing to 24, 48, and 72 hours beyond the normal 20-day gestational period. Between six and nine fetuses were harvested at all time points in both treatment and control groups. The morphology and cell biology of the spheno-occipital synchondroses, in constrained fetuses and unconstrained controls, were examined using hematoxylin and eosin-stained sections. Chondrocyte apoptosis was examined using terminal deoxynucleotidyl transferase-mediated dUDP end-labeling assays and electron microscopy. RESULTS: In nonconstrained animals, the spheno-occipital synchondrosis demonstrated normal architecture and normal chondrocyte morphology at all time points. In contrast, intrauterine constraint resulted in a progressive disruption of the normal cellular architecture of the spheno occipital synchondrosis over 72 hours, with premature ossification of the synchondrosis. Widespread chondrocyte apoptosis within the synchondrosial growth center was demonstrated by terminal deoxynucleotidyl transferase-mediated dUDP end-labeling assays and electron microscopy. CONCLUSION: These experiments confirm the ability of intrauterine constraint to induce changes in the morphology and cell biology of the cranial base in synostotic fetuses. PMID- 16217479 TI - Expression and possible mechanisms of regulation of BMP3 in rat cranial sutures. AB - BACKGROUND: Clinical genetics data and investigative studies have contributed greatly to our understanding of the role of numerous genes in craniosynostosis. Recent studies have introduced antagonists of osteogenesis as potential key regulators of suture fusion and patency. The authors investigated the expression pattern of the bone morphogenetic protein antagonist BMP3 in rat cranial sutures and the factors regulating its expression in vitro. METHODS: Microarray analysis was performed on rat posterior frontal and sagittal cranial sutures at 5, 10, 15, 20, and 30 days of life (n = 30 per group). Gene expression was confirmed using quantitative real-time reverse transcriptase polymerase chain reaction. Regulation of BMP3 expression was determined using primary rat calvarial osteoblasts stimulated with recombinant human fibroblast growth factor 2 or recombinant human transforming growth factor beta1, or cultured with primary rat nonsuture dura mater. Gene expression was quantified with quantitative real-time reverse transcriptase polymerase chain reaction. RESULTS: BMP3 expression in the posterior frontal suture decreased over the time course analyzed, whereas it increased in the sagittal suture. Notably, BMP3 expression was higher in the patent sagittal suture during the window of posterior frontal suture fusion. Stimulation of osteoblasts with recombinant human fibroblast growth factor 2 led to a rapid and sustained suppression of BMP3 expression (85 percent, p < 0.01) when compared with controls. Co-culture with dural cells decreased BMP3 mRNA by 50 percent compared with controls (p < 0.01). CONCLUSIONS: BMP3 is expressed in rat cranial sutures in a temporal pattern suggesting involvement in cranial suture patency and fusion. Furthermore, BMP3 is regulated in calvarial osteoblasts by recombinant human fibroblast growth factor 2 and by paracrine signaling from dura mater. These data add to our knowledge of the role of osteogenic antagonists in cranial suture biology. PMID- 16217481 TI - Use of the transforming growth factor-beta1 inhibitor peptide in periprosthetic capsular fibrosis: experimental model with tetraglycerol dipalmitate. AB - BACKGROUND: Capsular contracture is the most common specific complication associated with silicone prostheses. It is thought to be caused by the gradual retraction of the fibrous scar tissue that forms around the prosthesis. Molecular biology has made it possible to determine the role of transforming growth factor beta (TGF-beta) in the scar physiopathology of any fibrotic process, including periprosthetic capsular fibrosis. The effects on the inhibition of TGF-beta have also been demonstrated in experimental models of scar formation and fibrosis, which opens the way for new therapeutic alternatives in the treatment of capsular contracture. METHODS: Three experimental groups of 10 rats each were formed to evaluate periprosthetic fibrosis after its modulation with a newly synthesized TGF-beta1 peptide inhibitor in a tetraglycerol dipalmitate matrix. In the first group, subcutaneously and submuscularly placed, smooth, solid silicone prostheses were left untreated; in the second group, the prostheses were left after being immersed in a solution of tetraglycerol dipalmitate; and in the third group, following the same protocol as in the second group, the solution contained tetra glycerol dipalmitate mixed with the inhibitor peptide of TGF-beta1. The animals were euthanized 8 weeks after implantation, and the capsules were assessed both macroscopically and histologically. RESULTS: Inhibition of capsular thickness and cellularity was significantly more effective in the group of animals treated with the inhibitor peptide of TGF-beta1. CONCLUSIONS: The TGF-beta1 inhibitor peptide applied in a matrix with tetraglycerol dipalmitate is significantly effective in achieving a reduction in periprosthetic fibrosis after placement of silicone implants, either subcutaneously or submuscularly. This result suggests new therapeutic approaches. PMID- 16217482 TI - Differentiation-dependent up-regulation of BMP-2, TGF-beta1, and VEGF expression by FGF-2 in human bone marrow stromal cells. AB - BACKGROUND: Bone tissue formation by bone marrow stromal cells may be supported and enhanced by multiple growth factors, particularly in cases of a compromised local microenvironment. In this study, the authors hypothesized that fibroblast growth factor (FGF)-2 can stimulate the production by human bone marrow stromal cells of osteogenic [i.e., bone morphogenetic protein (BMP)-2 and transforming growth factor (TGF)-beta1] and angiogenic [i.e., vascular endothelial growth factor (VEGF)] factors. METHODS: Human bone marrow stromal cells from six donors were expanded for two passages (expansion phase) and subsequently cultivated in osteogenic medium containing ascorbic acid, beta-glycerophosphate, and dexamethasone (differentiation phase). After each phase, cells were transferred into serum-free medium with or without FGF-2 at different concentrations and for different times, and the expression of BMP-2, TGF-beta1, and VEGF was quantified at the mRNA level by real-time quantitative reverse-transcriptase polymerase chain reaction. The amounts of TGF-beta1 and VEGF released in the culture medium were assessed using enzyme-linked immunosorbent assay kits and normalized to the DNA content. RESULTS: In response to 5 ng/ml FGF-2 for 24 hours, the mRNA expression of VEGF increased at both culture phases (up to 6.1 fold), whereas that of BMP-2 and TGF-beta1 significantly increased only after the expansion (3.1 fold) or differentiation phase (2.1-fold), respectively. Similar trends were observed in the amounts of proteins measured in the culture medium. CONCLUSIONS: The authors' results indicate that FGF-2 up-regulates the expression of BMP-2, TGF-beta1, and VEGF in human bone marrow stromal cells, in a pattern dependent on the cell-differentiation stage. These findings prompt for in vivo investigations on the delivery of FGF-2 for the temporally/functionally regulated enhancement of bone marrow stromal cell-based bone induction. PMID- 16217483 TI - Hypertrophic scar fibroblasts have increased connective tissue growth factor expression after transforming growth factor-beta stimulation. AB - BACKGROUND: Hypertrophic scars and keloids respond to dermal disruption with excessive collagen deposition and increased transforming growth factor (TFG)-beta expression. Connective tissue growth factor (CTGF) is a downstream mediator of TGF-beta activity that is associated with scar and fibrosis. The authors hypothesize that there is increased expression of CTGF by hypertrophic scar and keloid fibroblasts in response to TGF-beta stimulation. METHODS: Primary fibroblasts were isolated in culture from human hypertrophic scar (n = 2), keloid (n = 2), and normal skin (n = 2). After 18 hours of serum starvation, the cells were stimulated with 10 ng/ml of TGF-beta1, TGF-beta2, and TGF-beta3 for 24 hours. Quantitative real-time polymerase chain reaction was performed on extracted RNA samples to assay for CTGF mRNA expression. RESULTS: Baseline CTGF expression was increased 20-fold in unstimulated hypertrophic scar fibroblasts and 15-fold in keloid fibroblasts compared with normal fibroblasts. CTGF expression increased greater than 150-fold when stimulated with TGF-beta1 (p < 0.002) and greater than 100-fold when stimulated by TGF-beta2 or TGF-beta3 compared with normal fibroblasts (p < 0.02 and p < 0.002, respectively). CTGF expression was greatest after TGF-beta1 stimulation in hypertrophic scar fibroblasts compared with TGF-beta2 (p < 0.04) and TGF-beta3 (p < 0.02). Keloid fibroblast CTGF expression also increased greater than 100-fold after stimulation with TGF-beta1 (p = 0.16) and greater than 75-fold after addition of TGF-beta2 and TGF-beta3 (p = 0.06 and p = 0.22, respectively). CONCLUSIONS: Hypertrophic scar fibroblasts have both intrinsic up-regulation of CTGF transcription and an exaggerated capacity for CTGF transcription in response to TGF-beta stimulation. These data suggest that blockage of CTGF activity may reduce pathologic scar formation. PMID- 16217485 TI - Microarray analysis of mechanical shear effects on flexor tendon cells. AB - BACKGROUND: Adhesion formation after flexor tendon repair remains a clinical problem. Early postoperative motion after tendon repair has been demonstrated to reduce adhesion formation while increasing tendon strength. The authors hypothesized that during mobilization, tendon cells experience mechanical shear forces that alter their biology in a fashion that reduces scar formation but also activates key genes involved in tendon healing. METHODS: To test this hypothesis, primary intrinsic tenocyte cultures were established from flexor tendons of 20 Sprague-Dawley rats and sheared at 50 rpm (0.41 Pa) using a cone viscometer for 6 and 12 hours. Total RNA was harvested and compared with time-matched unsheared controls using cDNA microarrays and Northern blot analysis. RESULTS: Microarray analysis demonstrated that mechanical shear stress induced an overall "antifibrotic" expression pattern with decreased transcription of collagen type I and collagen type III. Shear stress down-regulated profibrotic molecules in the platelet-derived growth factor, insulin-like growth factor, and fibroblast growth factor signaling pathways. In addition, shear stress induced an overall decrease in transforming growth factor (TGF)-beta signaling pathway molecules with down regulation of TGF-beta2, TGF-beta3, TGF-RI, and TGF-RII expression. Moreover, sheared tendon cells increased expression of matrix metalloproteinases and decreased expression of tissue inhibitors of metalloproteinase, an expression pattern consistent with an antifibrotic increase in extracellular matrix degradation. However, the authors also found up-regulation of genes implicated in tendon healing, specifically, vascular endothelial growth factor-A and several bone morphogenetic proteins. Interestingly, the known mechanoresponsive gene, TGF beta1, also implicated in tendon healing, was differentially up-regulated by shear stress. Northern blot validation of our results for TGF-beta1, TGF-beta2, TGF-beta3, and collagen type I demonstrated direct correlation with the authors' microarray data. CONCLUSIONS: The authors demonstrate an overall antifibrotic expression pattern in response to shear stress in tendon cells that may provide insight into the mechanisms by which early mobilization decreases adhesion formation without impaired tendon healing. PMID- 16217487 TI - Increased cutaneous nerve fibers in female specimens. AB - BACKGROUND: The control of pain is critical to performing surgery. All surgical procedures cause some degree of pain, and the ability to minimize pain often affects a patient's perception of surgical outcome. Although the development of surgery was boosted by the advent of anesthesia, inadequate pain control continues to plague modern medicine. The mechanism of pain induction is an important area of research in the health care industry. To date, few studies have demonstrated increased perception of pain and lower tolerance for pain in female patients when compared with male patients. The authors hypothesized about whether these differences were related to increased density of nerve fibers in female as compared with male patients. METHODS: The density of nerve fibers at a specific location (the skin directly overlying the infraorbital nerve foramen) was measured to test this hypothesis. Twenty cadaver skin specimens (1 cm2) were harvested, prepared using immunohistochemistry (S-100 polyclonal antibody), and counted using 45x high-powered microscopy. RESULTS: Female specimens (n = 10) demonstrated increased nerve fiber density (34 +/- 19 fibers/cm skin) when compared with male specimens (n = 10; 17 +/- 8 fibers/cm skin; p = 0.038). CONCLUSION: Although preliminary and limited in scope, these findings favor a physical (organic) rather than a psychosocial explanation for more pronounced pain perception in female patients. PMID- 16217488 TI - Nonsurgical delay of dorsal rat cutaneous flap using a long-pulsed 1064-nm Nd:YAG laser with a contact cooling device. AB - BACKGROUND: This study evaluated the efficiency of a long-pulsed neodymium:yttrium-aluminum-garnet laser, operating at 1064 nm and equipped with a contact cooling device, in the delay of a caudally based dorsal rat skin flap (10 x 3 cm). This laser has deeper tissue penetration and has not been used for this purpose before. METHODS: Twelve male Sprague-Dawley rats were used in each of six groups. The delay effects of three different laser treatment patterns (only longitudinal borders, cephalic and longitudinal borders, and entire surface of the 10 x 3-cm flap) were compared with an acute untreated control flap as well as two surgical delay methods (incision of longitudinal borders and incision of longitudinal borders plus flap undermining). The laser effects on the cutaneous vasculature and perfusion were assessed by intravenous fluorescein injection, histologic study, microangiography, and in vivo real-time video monitoring. RESULTS: Selective thermocoagulation of subdermal vessels was achieved using a 6 mm spot, 140-J/cm fluence, and 40-msec pulse width. In the cephalic and longitudinal borders laser-treated group, a delay effect was achieved. The maximum delay effect was achieved by the surgical delay group that used the method of incision of the longitudinal borders plus flap undermining. Laser treatment of only the longitudinal borders did not improve flap survival, whereas treatment of the entire flap surface significantly reduced flap survival. CONCLUSION: Nonsurgical delay of a dorsal rat cutaneous flap is possible by selective occlusion of the subdermal plexus at the longitudinal and cephalic borders of the planned flap using a long-pulsed 1064-nm neodymium:yttrium aluminum-garnet laser equipped with a contact cooling device. PMID- 16217489 TI - Cheek and eyelid reconstruction: the resurrection of the angle rotation flap. AB - BACKGROUND: Reconstruction of larger cheek and eyelid defects may pose a dilemma for surgeons, since flaps used in reconstruction may be difficult to design, be unreliable, require extensive dissection, and result in neck scarring. Consequently, the authors wish to simplify and expand an overlooked flap, the angle rotation flap, which moves tissue in both a medial and upward direction. METHODS: Twenty patients with cheek and eyelid defects were treated by the angle rotation flap. In this flap design, the angle designed below the ear was closed primarily and the neck tissue previously there was rotated upward and forward to lie in front of the ear. The portion of the flap that was in front of the ear was transposed to the lower lid/cheek area. This flap was modified in several patients by elevation in the deep plane and first-stage tissue expansion. RESULTS: In all cases, good coverage was provided for medial cheek and lower eyelid defects with minimal scarring on the neck. There were no flap losses of any kind. There were no major complications, and all minor incidences were treated by minimal procedures without long-term sequelae. CONCLUSION: The modified angle rotation flap is a useful tool for cheek and eyelid defects. PMID- 16217491 TI - Free partial medial rectus muscle flap for closure of complex extremity wounds. PMID- 16217492 TI - Temporary assisting suspension suture technique for successful microvascular anastomosis of extremely small and thin walled vessels for mice transplantation surgery. AB - Dissection and microsurgical anastomosis in small and thin-walled vessels is challenging. Temporary assisting suspension suture technique was developed to overcome those difficulties in establishing successful composite tissue allotransplantation in mice. The operations were performed in 12- to 16-week-old Balb/c mice weighing 25 to 30 grams as both donor and recipient animals. Extended vascularized groin cutaneous flaps based on the superficial epigastric vessels were used. A total of 10 groin cutaneous flaps were transplanted. Three temporary assisting suspension sutures of 11-0 nylon were placed at the 12-, 4-, and 8 o'clock positions to donor and recipient artery and vein before the anastomosis. This technique allowed atraumatic dissection of delicate and thin vessels, prevented vessel wall collapse, and facilitated adequate exposure of the lumen during placement of the permanent microvascular sutures. Thus, the microvascular anastomosis was performed in an unusual manner. The temporary assisting suspension sutures were removed just before the permanent suture was tied down. The mean operation time was 1 hour and 45 minutes with an ischemia time of 1 hour. Ninety-percent success in immediate and late-term patency rates was achieved, which was confirmed by transplant survival. This technique was proven to be useful for microvascular anastomosis in thin-walled vessels and is recommended. PMID- 16217493 TI - Orthognathic surgery: is there a future? AB - BACKGROUND: Despite significant clinical advances in orthognathic surgery, a perceived decrease in recent years in the number of orthognathic cases was noted at the authors' institution. Word of mouth suggested that this phenomenon was region-wide. To explore this possible dichotomy, a one-page questionnaire was designed and sent to all plastic surgeons in the American Society of Maxillofacial Surgeons and to all active practicing oral surgeons and orthodontists in the state of Ohio. METHODS: The questionnaire was designed to estimate both the surgeons' and the orthodontists' perception of whether the number of orthognathic cases was increasing, decreasing, or remaining the same over a 5-year period (from 1996 to 2000). A second questionnaire was then sent only to plastic surgeons and oral surgeons to further evaluate reasons for this possible increase or decrease. RESULTS: The overall response to the first survey was 39 percent (236 of 601 surveys sent); 87 responses (32 percent) were from plastic and oral surgeons, and 132 (40 percent) were from orthodontists. Seventy five percent of responders perceived a decrease in the number of orthognathic operations. Of those, more than 80 percent claimed that reimbursement was the major reason for the reduction in the number of surgical procedures. The degree of reduction in reimbursement over this 5-year period was documented at the authors' institution, and the perceived reduction over this 5-year period was estimated by the respondents. Finally, reimbursement per hour was calculated for orthognathic cases and compared with three other standard plastic surgery procedures. CONCLUSION: Implications of this reduction in orthognathic surgery in Ohio are discussed, and suggestions for better documentation of this health care issue are suggested. PMID- 16217495 TI - The potential of adipose-derived adult stem cells as a source of neuronal progenitor cells. AB - Adipose-derived adult stem cells are a population of mesenchymal stem cells extracted from discarded adipose tissue. Many have reported the differentiation of adipose-derived stem cells into chondrocytes, myocytes, osteoblasts, and, most recently, neural progenitor cells. This article covers the current state of the potential of the differentiation of adipose-derived stem cells into neuronal cells and an overview of their potential as adult stem therapies for neurological disorders. It has been reported that adipose-derived stem cells are capable of undergoing neuronal differentiation using protocols similar to that of Woodbury et al., which reported the differentiation of bone marrow stromal cells specifically into neurons. However, the transdifferentiation of bone marrow stromal cells into neuronal cells has recently fallen under intense criticism, which will likely place the plasticity of adipose-derived stem cells under scrutiny as well. To date, no group has produced evidence that adipose-derived stem cells are capable of differentiating to mature, functional neuronal cells in vitro. However, recent in vivo studies with adipose-derived stem cells are promising. PMID- 16217496 TI - Who benefits from peer review? An analysis of the outcome of 100 requests for review by Plastic and Reconstructive Surgery. AB - BACKGROUND: Little is known of what is done with the comments on submitted manuscripts provided by peer reviewers or to what extent these comments benefit the editor in deciding to accept or reject the manuscript, the author(s) in revising their manuscript, or the readership at large. Furthermore, nothing is known of any possible benefits of the process to the peer reviewer. Finally, the peer-review process may even be maleficent because of its implicit delay of publication and a possible bias against manuscripts originating from non-Anglo American countries. METHODS: The authors evaluated the benefits of the peer review process to authors, editor, readers, and reviewers by a bibliometric analysis of the outcome of 100 requests for review made by the editor of Plastic and Reconstructive Surgery from 1992 through 2003. The publication delay and potential geographical bias were evaluated as potential disadvantages. RESULTS: The authors' reviewer advised acceptance of 56 percent of the manuscripts, and the editor mostly agreed with his advice. This suggests that the editor benefited from the review. The authors addressed 48 to 81 percent of the reviewer's constructive suggestions, and this suggests that they and the readers benefited also. Readers of Plastic and Reconstructive Surgery may further benefit because manuscripts rejected by Plastic and Reconstructive Surgery end up in less prestigious journals. The implicit delay of publication is limited, and the authors found no bias against non-Anglo-American submissions. The cost effectiveness of the process for the peer reviewer remains unclear. CONCLUSIONS: The peer-review system of Plastic and Reconstructive Surgery, in general, is beneficial. PMID- 16217498 TI - An anatomical comparison of transpalpebral, endoscopic, and coronal approaches to demonstrate exposure and extent of brow depressor muscle resection. AB - BACKGROUND: Approaches for exposure of the muscles of brow depression include transpalpebral, endoscopically assisted, and open coronal techniques. The purpose of this anatomical study was to compare the capacity for visualization and amount of brow depressor muscle resection with each technique. METHODS: The corrugator supercilii, depressor supercilii, medial orbicularis oculi, and procerus muscles were studied by gross anatomical dissection carried out on 24 sides of 12 cadaver heads. First, all visible corrugator and depressor supercilii muscles were resected by means of an upper blepharoplasty incision. Subsequently, a subgaleal endoscopic approach was used to evaluate the extent of resection performed and remove the remaining muscle after transpalpebral corrugator resection. This was followed by coronal exposure to assess the anatomical location and extent of muscle resection accomplished by the two previously mentioned techniques. RESULTS: In all dissections, endoscopy demonstrated that up to one-third of the lateral aspect of the transverse heads of the corrugator supercilii remained following transpalpebral resection. Oblique corrugator head resections were complete, without significant residual muscle following transpalpebral corrugator resection. The procerus muscle was able to be blindly transected by means of the transpalpebral incision but exposed and ablated in all 12 specimens using endoscopy. Coronal exposure demonstrated that no significant amount of corrugator, depressor supercilii, or procerus muscle remained in any of the 12 heads following endoscopically assisted exposure and resection. The medial head of the orbital portion of the orbicularis oculi was visualized and accessible regardless of the technique used. CONCLUSIONS: In 24 anatomical dissections, transpalpebral corrugator resection failed to remove up to one-third of the transverse head of the corrugator muscle. Removal of the brow depressor muscles was accomplished with the endoscopic approach, as confirmed by coronal exposure. PMID- 16217500 TI - Mesh lift: a new procedure for long-lasting results in brow lift surgery. AB - BACKGROUND: Prevention of recurrent eyebrow ptosis is the greatest challenge in brow lift surgery today. In this article, the author describes a new surgical procedure to provide long-lasting results in brow lift surgery. METHODS: Over 7 years, this new technique was used in 37 patients. Except for four, all patients were women aged 22 to 57 years. In this procedure, a polypropylene mesh strap is used as a suspender to maintain the elevated position of the eyebrow. The mesh suspender is placed in a subgaleal tunnel that is created between a limited temporal scalp incision and a classic upper blepharoplasty incision. The distal end of the mesh is sutured to the undersurface of the upper orbital part of the orbicularis oculi muscle, and the proximal end is fixed to the periosteum at the temporal region after a desired eyebrow position is obtained by traction of the mesh suspender superolaterally. RESULTS: The polypropylene mesh was tolerated well in all patients. No complication related to foreign body reaction was encountered. During 6 months to 4 years of follow-up, none of the patients experienced recurrent eyebrow ptosis. The long-term results revealed a long lasting, almost permanent eyebrow elevation in all patients. CONCLUSIONS: This new technique seems to be useful in prevention of recurrent eyebrow ptosis following brow lift surgery. The author suggests that, after its integration with reoperative tissue, the mesh suspender becomes an artificial suspensory aponeurosis that provides stable fixation to maintain the elevated position of the eyebrow. This is considered to be the main reason for the excellent long lasting results obtained in the current clinical study. PMID- 16217502 TI - The use of alloplastic materials in secondary rhinoplasties: 32 years of clinical experience. AB - BACKGROUND: A retrospective evaluation of the authors' 32 years of experience in revision rhinoplasty is presented. The authors suggest that iatrogenic nasal deformities are studied under four groups on the basis of their location and the affected structures: lower third, middle third, upper third, and combined deformities. The authors also present the vertical columellar incision for insertion of alloplastic implants and cartilage grafts, a genuine approach avoiding contamination with the nasal flora. METHODS: A total of 182 cases were studied, as follows: lower third (n = 81), middle third (n = 65), upper third (n = 17), and combined (n = 19) deformities. The mean age of the patients was 40 years. RESULTS: Our follow-up was a minimum of 2 years, and some of the cases with Proplast implants have been followed for as long as 21 years. During this rather long follow-up, only two of the Proplast implants had to be removed (one because of an acute infection and the other because of a chronic infection causing extrusion after 5 years). CONCLUSIONS: The authors have always preferred to use autogenous cartilage grafts for tip deformities and Proplast implants for middle and upper third saddle nose deformities whenever necessary. Their long term results with Proplast for more than 20 years show that alloplastic materials may be as reliable as autogenous implants if the surgical principles are met and the cases are carefully selected. PMID- 16217503 TI - Components of the short nostril. AB - BACKGROUND: The short nostril, best visualized on the basilar view, is a multifaceted dysmorphology that requires evaluation beyond that of alar/columellar deformities. While the soft triangle is the key component in short nostril disharmony, the alar rim and cartilaginous structures that border the nostrils play a salient role as well. METHODS: A retrospective review of 200 consecutive rhinoplasties (primary and secondary) examined the specific role of soft triangle excision and other components in the short nostril deformity. Twenty-seven patients underwent soft triangle excision with or without alteration of the other structures influential on nostril length. Of these 27 patients, only three patients required soft triangle excision alone. RESULTS: The distance from the nostril apex to the caudal border of the alar dome was found to be the crucial element in defining the treatment approach for creating nostril length. When this distance was long, excision of the soft triangle lining and approximation of the alar rim to the lining under the dome elevated the nostril apex and elongated the nostril. When the distance between the nostril apex and overlying dome was ideal or short, soft triangle lining removal was not required, and an optimal nostril length was established by repositioning the other components. Raising the dome using transdomal sutures redirected the wide domal arch vertically, narrowing and lengthening the nostril, provided there was no redundancy in the soft triangle. In a similar fashion, interdomal sutures improved both nostril length and inclination. Placement of a columellar strut also elongated the nostril. An alar rim graft, used primarily to correct alar rim retraction and concavity, also elongated the short nostril. CONCLUSIONS: The most important factor in analysis and treatment of the short nostril is the extent of the soft triangle tissue present. Soft triangle lining removal is indicated when the distance from the nostril apex to the caudal dome is excessive. This allows the nostril apex to be pulled anteriorly, thus elongating the nostril. The short nostril often coexists with multiple other abnormalities of the nasal base and tip, mandating a comprehensive approach to address all the deformities encountered. Correction of alar retraction also effectively increases nostril length. Further improvement of asymmetric tips and nostrils can be achieved through unilateral soft triangle lining excision with dome equalization through tip suturing and a subdomal graft. PMID- 16217504 TI - Mini spreader grafts: a new technique associated with reshaping of the nasal tip. AB - BACKGROUND: Reshaping of the nasal tip by excision of a strip of cartilage from the cephalic portion of the lateral crura is a commonly used procedure. This surgical technique is not completely free of risk, above all when associated with hump excision, as is often the case. It can lead to incompetence of the internal nasal valve and thus to postoperative respiratory disorders. METHODS: The author proposes a new surgical technique capable of preventing these problems and improving the degree of nasal tip rotation, where necessary. The method combines resection of the cephalic portion of the lateral crura with its attachment on either side of the dorsal septum as two "mini spreader grafts." Forty patients were treated by means of this technique. RESULTS: Rhinomanometric data demonstrate an improvement in total mean nasal airflow, and no worsening was registered in any case. Correction of droopy tip was also achieved with optimal cephalic rotation of the tip. CONCLUSIONS: Although small in size, the grafts are very important in functional terms in that they restore a physiologic angle in the area of the internal nasal valve between the septum and the upper lateral cartilages. The preservation of a fibromucous connection between the grafts and the two lateral crura from which they are taken has an aesthetic effect. To this end, it is sufficient to secure the two mini spreader grafts to the septum in a more cephalic position to obtain a greater degree of rotation of the tip complex. PMID- 16217505 TI - A classification of contour deformities after bariatric weight loss: the Pittsburgh Rating Scale. AB - BACKGROUND: Contour deformities after massive weight loss are diverse and often severe in nature. Current progress has necessitated a valid, accessible, and comprehensive rating system that correlates appearance and appropriate surgical treatment. Presently, no existing rating system addresses the breadth and variety of deformities that can occur or allows for adequate postsurgical evaluation. METHODS: The authors reviewed full-body photographs of over 300 female patients seen between October of 2002 and May of 2004. The authors targeted body areas most frequently demonstrating skin and soft-tissue laxity and ptosis. A 10 region, four-point grading system was designed to describe the common deformities found in each region of the body. To validate the scale, 12 trained observers applied the rating scale to photographs of 25 patients who showed the 10 regions. Each grading scale ranged from 0, indicating normal, to 3, indicating the most severe deformity. Repeat testing was performed at 2 weeks. Interobserver validity and test-retest reliability were determined using weighted kappa analysis. RESULTS: In all 10 categories, the kappa value was 0.6 or higher (0.6 = threshold for good validity), with a mean kappa value of 0.68 (range, 0.61 to 0.78) and an overall agreement of 69 percent over two sessions. All 12 observers scored an individual mean kappa value of greater than 0.6, indicating good interobserver validity. A given observer had a mean 67 percent agreement, indicating reasonable test-retest reliability. CONCLUSIONS: The Pittsburgh Rating Scale is a validated measure of contour deformities after bariatric weight loss. This scale may have applications in preoperative planning and evaluating surgical outcomes. PMID- 16217507 TI - Show-and-tell: some things never change. PMID- 16217508 TI - Hurricane Katrina, plastic surgery, and you: the art of giving back. PMID- 16217509 TI - Formal decision trees in plastic surgery. PMID- 16217513 TI - Extended lower trapezius island myocutaneous flap in the repair of postburn axillary contracture. PMID- 16217515 TI - Postsurgical eyelid position. PMID- 16217516 TI - Reconstruction in women with bilateral prophylactic mastectomy: a descriptive study. PMID- 16217517 TI - Vascularized fibula graft and osseointegrated teeth implantation after segmental mandibulectomy for fibrous dysplasia. PMID- 16217518 TI - Liposuction breast reduction. PMID- 16217520 TI - Terminolateral fiber count. PMID- 16217521 TI - Concerns over the use of tissue adhesive in facial lacerations: recommendations and a suggestion to increase lag bond strength. PMID- 16217523 TI - Preliminary results after face peels using Aldara cream 5% (imiquimod). PMID- 16217524 TI - Using a roller-bottle filled with adhesive solution to fix silk tape on the face. PMID- 16217525 TI - Marriage of autogenous bone and hydroxyapatite cement for reconstruction of frontal bone defect. PMID- 16217526 TI - Regional bone flap for mandibular reconstruction. PMID- 16217527 TI - A simple mouth-opening device. PMID- 16217528 TI - Temporomandibular joint dislocation in an infant after vomiting. PMID- 16217529 TI - Angled blade for cleft palate surgery. PMID- 16217530 TI - Disposable nipple marker. PMID- 16217531 TI - A simple technique for meshing long split skin grafts. PMID- 16217532 TI - Consult for exposed hardware. PMID- 16217534 TI - Another use for a measuring caliper. PMID- 16217533 TI - Treatment of hidradenitis suppurativa with botulinum toxin A. PMID- 16217535 TI - Further uses of a simple syringe. PMID- 16217536 TI - A novel method for seroma drainage. PMID- 16217537 TI - Atypical location of chickenpox vesicular lesions after fingertip injury. PMID- 16217538 TI - Telfa as donor-site dressing. PMID- 16217539 TI - A simple, effective, and liberating method to protect casts and splints. PMID- 16217540 TI - Malignant blue nevus: a confusing diagnosis for the surgeon. PMID- 16217541 TI - Use of a newspaper for a splint in emergency settings. PMID- 16217542 TI - The palm as a holster. PMID- 16217543 TI - [Amoebic infections of the lung and pleura]. AB - INTRODUCTION: Pleuropulmonary involvement is the second most common extra intestinal manifestation of entamoeba histolytica infection after liver abscess. CASE REPORT: We report 2 cases of pleuropulmonary disease occurring in two men aged 32 and 48 years following an episode of dysentery. Purulent pleural infection was noted in one case. In the other both lung and liver abscesses occurred. CONCLUSION: The diagnosis was confirmed by strongly positive serology in both cases. Treatment with metronidazole (1.5 g per day) for 15 days combined with pleural drainage led to a satisfactory outcome in both cases. PMID- 16217544 TI - [Mediastinal lymph node dissection: curative procedure or Will Rogers phenomenom only]. PMID- 16217545 TI - [Francophonie and multilingualism]. PMID- 16217546 TI - [Rare and orphan diseases--a national priority]. PMID- 16217547 TI - Gain and loss of multiple genes during the evolution of Helicobacter pylori. AB - Sequence diversity and gene content distinguish most isolates of Helicobacter pylori. Even greater sequence differences differentiate distinct populations of H. pylori from different continents, but it was not clear whether these populations also differ in gene content. To address this question, we tested 56 globally representative strains of H. pylori and four strains of Helicobacter acinonychis with whole genome microarrays. Of the weighted average of 1,531 genes present in the two sequenced genomes, 25% are absent in at least one strain of H. pylori and 21% were absent or variable in H. acinonychis. We extrapolate that the core genome present in all isolates of H. pylori contains 1,111 genes. Variable genes tend to be small and possess unusual GC content; many of them have probably been imported by horizontal gene transfer. Phylogenetic trees based on the microarray data differ from those based on sequences of seven genes from the core genome. These discrepancies are due to homoplasies resulting from independent gene loss by deletion or recombination in multiple strains, which distort phylogenetic patterns. The patterns of these discrepancies versus population structure allow a reconstruction of the timing of the acquisition of variable genes within this species. Variable genes that are located within the cag pathogenicity island were apparently first acquired en bloc after speciation. In contrast, most other variable genes are of unknown function or encode restriction/modification enzymes, transposases, or outer membrane proteins. These seem to have been acquired prior to speciation of H. pylori and were subsequently lost by convergent evolution within individual strains. Thus, the use of microarrays can reveal patterns of gene gain or loss when examined within a phylogenetic context that is based on sequences of core genes. PMID- 16217549 TI - 31P MRS analysis of the phospholipid composition of normal human peripheral blood mononuclear cells (PBMC). AB - The aim of this investigation was to characterize the phospholipid composition of normal human blood mononuclear cells using 31P NMR spectroscopy. Mononuclear cells of peripheral blood were obtained from 10 volunteers. Phospholipid extracts were prepared from 60x10(6) cells according to modified Folch's method. An AMX 300 Bruker spectrometer 7.05 T was used. The 31P spectrum of phospholipid extracts from normal human PBMC consisted of 9 peaks, with one each for phosphatidylcholine (PC), plasmalogen of phosphatidylcholine (CPLAS), lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and cardiolipin (CL), and another one due to the external reference substance, methylenediphosphonic acid (MDPA). The concentrations of these phospholipids (PL), based on the integral intensities, were as follows: 0.398 +/- 0.078 mmole/l for PC; 0.033 +/- 0.019 mmole/l for CPLAS; 0.155 +/- 0.043 mmole/l for SM; 0.266 +/- 0.104 mmole/l for PI+PE; 0.101 +/- 0.040 mmole/l for PS, and 0.026 +/- 0.033 mmole/l for CL. The results of this study confirmed that 31P MRS is a convenient tool for measuring the phospholipid concentrations of biological samples. PMID- 16217550 TI - Enhanced production of isoamyl alcohol and isoamyl acetate by ubiquitination deficient Saccharomyces cerevisiae mutants. AB - Aromatic compounds are an important element in the flavor of yeast-fermented alcohol. We isolated mutants of Saccharomyces cerevisiae capable of growth at high levels of hydrostatic pressure. Among them, the HPG1 mutants, with a defect in their Rsp5 ubiquitin ligase, were found to produce high amounts of aromatics due to enhanced leucine uptake, with isoamyl alcohol production 2- to 3-fold and isoamyl acetate production 4- to 8-fold that of the wild-type strain. The result suggests that the HPG1/RSP5 mutant alleles could be new resources for producing these flavoring compounds for yeast-fermented alcoholic beverages. PMID- 16217548 TI - Protein molecular function prediction by Bayesian phylogenomics. AB - We present a statistical graphical model to infer specific molecular function for unannotated protein sequences using homology. Based on phylogenomic principles, SIFTER (Statistical Inference of Function Through Evolutionary Relationships) accurately predicts molecular function for members of a protein family given a reconciled phylogeny and available function annotations, even when the data are sparse or noisy. Our method produced specific and consistent molecular function predictions across 100 Pfam families in comparison to the Gene Ontology annotation database, BLAST, GOtcha, and Orthostrapper. We performed a more detailed exploration of functional predictions on the adenosine-5' monophosphate/adenosine deaminase family and the lactate/malate dehydrogenase family, in the former case comparing the predictions against a gold standard set of published functional characterizations. Given function annotations for 3% of the proteins in the deaminase family, SIFTER achieves 96% accuracy in predicting molecular function for experimentally characterized proteins as reported in the literature. The accuracy of SIFTER on this dataset is a significant improvement over other currently available methods such as BLAST (75%), GeneQuiz (64%), GOtcha (89%), and Orthostrapper (11%). We also experimentally characterized the adenosine deaminase from Plasmodium falciparum, confirming SIFTER's prediction. The results illustrate the predictive power of exploiting a statistical model of function evolution in phylogenomic problems. A software implementation of SIFTER is available from the authors. PMID- 16217551 TI - The influence of methylene blue on the spontaneous contractity of the non pregnant human myometrium and on the myometrial response to DEA/NO. AB - Nitric oxide (NO) is a potent inhibitor of spontaneous contractions of the human non-pregnant myometrium; however, the precise mechanism by which NO causes the myometrial smooth muscles to relax remains unclear. The aim of this study was to determine the influence of methylene blue (MB) on myometrial contractions and the response of the myometrium to DEA/NO in vitro. Concentration-response curves to DEA/NO were constructed in the absence and presence of MB (5x10(-6), 10(-4) and 10(-2) mol/l) and 5x10(-3) mol/l cystamine. Cystamine did not counteract the DEA/NO-induced relaxation of the myometrial strips. MB itself, excluding the lowest concentration, caused noticeable changes in spontaneous activity. The changes involved a concentration-dependent increase in the frequency of contractions, and a decrease in their amplitude. In conclusion, our results confirm that NO relaxes the human myometrium via a cGMP-independent mechanism. The results obtained in the presence of MB may be misleading because of its complex influence on myometrial contractile activity. PMID- 16217552 TI - Cloning and characterization of cDNA for the Oryza sativa phosphate transporter. AB - A putative high-affinity phosphate (Pi) transporter gene in rice (Oryza sativa), OsLPT1, was isolated by RT-PCR from the leaves of the plants. The 1635-bp nucleotide sequence of OsLPT1 spans an open reading frame encoding a polypeptide of 535 amino acids with sequence similarity to phosphate transporters from other plant species. Southern blot analysis showed that the OsLPT1 gene might be present in three transcripts in the rice genome. RT-PCR analysis demonstrated the expression of OsLPT1 in both leaves and roots. The expression of OsLPT1 in the roots was enhanced by Pi deprivation. In situ hybridization revealed OsLPT1 expression in mesophyll cells, xylem parenchyma and phloem cells in the leaves, and in the epidermis, exodermis, and in the vasculature surrounding metaxylem vessels in the roots. The data suggests that the OsLPT1 protein may be involved in enhancing phosphate uptake under conditions of Pi starvation, and in the translocation of Pi among cells in shoots to increase the efficiency of internal Pi use. PMID- 16217553 TI - Lead-induced cell death of human neuroblastoma cells involves GSH deprivation. AB - Lead (Pb2+) is a toxic heavy metal that has adverse effects on the health of humans and other animals. The developing central nervous system is especially sensitive and vulnerable to Pb2+ toxicity. In this study, the effects of low levels of Pb2+ exposure on human SH-SY5Y neuroblastoma cell cultures were assessed. The cells were exposed to Pb2+ (0.01 microM-10 microM) for 48 hrs, and the level of cell proliferation was determined. Pb2+ significantly inhibited the proliferation of neuroblastoma cells in a concentration-dependent manner. A 50% inhibition (IC50) in cellular proliferation was observed with 5 microM Pb2+. A significant decrease in the levels of glutathione (GSH), a critical intracellular antioxidant, was observed at all the lead concentrations. There was also a multifold increase in the activity of caspase-3, a key executioner of apoptosis, and in the levels of prostaglandin E2 (PGE2). Our results suggest that the neurotoxic effects of Pb may be mediated by apoptosis and PGE2 release, which could be potentially detrimental to neuronal survival. PMID- 16217554 TI - Cross-species bacterial artificial chromosome (BAC) library screening via overgo based hybridization and BAC-contig mapping of a yield enhancement quantitative trait locus (QTL) yld1.1 in the Malaysian wild rice Oryza rufipogon. AB - The construction of BAC-contig physical maps is an important step towards a partial or ultimate genome sequence analysis. Here, we describe our initial efforts to apply an overgo approach to screen a BAC library of the Malaysian wild rice species, Oryza rufipogon. Overgo design is based on repetitive element masking and sequence uniqueness, and uses short probes (approximately 40 bp), making this method highly efficient and specific. Pairs of 24-bp oligos that contain an 8-bp overlap were developed from the publicly available genomic sequences of the cultivated rice, O. sativa, to generate 20 overgo probes for a 1 Mb region that encompasses a yield enhancement QTL yld1.1 in O. rufipogon. The advantages of a high similarity in melting temperature, hybridization kinetics and specific activities of overgos further enabled a pooling strategy for library screening by filter hybridization. Two pools of ten overgos each were hybridized to high-density filters representing the O. rufipogon genomic BAC library. These screening tests succeeded in providing 69 PCR-verified positive hits from a total of 23,040 BAC clones of the entire O. rufipogon library. A minimal tilling path of clones was generated to contribute to a fully covered BAC-contig map of the targeted 1-Mb region. The developed protocol for overgo design based on O. sativa sequences as a comparative genomic framework, and the pooled overgo hybridization screening technique are suitable means for high-resolution physical mapping and the identification of BAC candidates for sequencing. PMID- 16217555 TI - The subcellular distribution of the p53 tumour suppressor, and organismal ageing. AB - The p53 protein, the product of a tumour suppressor gene, is a key regulator of cell growth, differentiation and apoptosis. It is able to induce a transient cell cycle arrest and terminal senescence. Most of its functions are exerted by the transcriptional activation of genes involved in cell cycle control, DNA repair and apoptosis. The activation of p53 is primarily mediated by post-translational modifications that affect its conformation and capacity to bind to several proteins, resulting in its stabilization and enhanced DNA-binding potential. Another way to regulate the biological function of p53 involves changes in its intracellular distribution. This paper presents an overview of the role of p53 in cellular senescence and the regulation of p53 activity by its intracellular distribution. PMID- 16217556 TI - Serum-resistant gene transfer to oral cancer cells by Metafectene and GeneJammer: application to HSV-tk/ganciclovir-mediated cytotoxicity. AB - Cationic lipids and polyamines have been used as non-viral gene transfer reagents, both in vitro and in vivo. One of the limitations to their use in vivo is the inhibition of gene delivery by serum. We showed previously that, in the absence of serum, relatively high cytotoxicity in oral cancer cell lines could be achieved via transfection of the Herpes Simplex Virus thymidine kinase (HSV-tk) gene followed by treatment with ganciclovir (GCV), despite the low efficiency of transfection (Konopka et al., Gene Ther. Mol. Biol. 8 (2004) 307-318). In this study we evaluated the effect of high concentrations (20-60%) of fetal bovine serum (FBS) on the transfection efficiency of two novel reagents, the polycationic liposome, Metafectene, and the polyamine reagent, GeneJammer, in HSC 3 and H357 human oral squamous cell carcinoma (OSCC) cells. We also examined whether the HSV-tk gene delivered in the presence of FBS (up to 60%, could induce cell death following treatment with GCV. Transfection was optimized using a luciferase-expressing plasmid. Both Metafectene- and GeneJammer-mediated luciferase gene expression in HSC-3 cells was reduced by 40-50% when transfection was performed in the presence of 20-60% FBS. The delivery of the HSV-tk gene by Metafectene in the absence and the presence of 60% FBS, followed by GCV treatment for 9 days, resulted in 95% and 70% cytotoxicity, respectively. With GeneJammer, transfection in 0% and 60% FBS resulted in 90% and 40% cytotoxicity, respectively, after 9 days. In contrast, very low transfection activity and a much higher inhibitory effect of serum were observed in H357 cells. Nevertheless, about 35% GCV-mediated cytotoxicity was observed with H357 cells at both 0% and 60% FBS, using GeneJamer. Thus, Metafectene and GeneJammer can be used in the delivery of genes in biological milieu and in the gene therapy of OSCC in animal models. PMID- 16217557 TI - Changes in the activity of acetylcholinesterase and Na,K-ATPase in human erythrocytes irradiated with X-rays. AB - The response of human erythrocytes to X-rays in the dose range from 40 Gy to 600 Gy was determined on the basis of changes in the activities of AChE and ATPase. The Na,K-ATPase activity increased above the control value at doses below 200 Gy, while at the doses higher than 200 Gy, it decreased, reaching 96% of the control value at a dose of 600 Gy. In the range of doses up to 200 Gy, the AChE activity, expressed as Vmax, did not change. At higher doses, it fell drastically, reaching 33% of the control value at a dose of 600 Gy. Simultaneously, the enzyme substrate affinity decreased at 200 Gy, and then started to increase at lower values of Vmax. The obtained results suggest that under appropriate conditions, low doses of radiation may have the opposite effects to high doses. PMID- 16217558 TI - The mammalian target of the rapamycin (mTOR) kinase pathway: its role in tumourigenesis and targeted antitumour therapy. AB - The mammalian target of rapamycin (mTOR) is a kinase responsible for mitogen induced cell proliferation/survival signaling. Its activation in response to mitogens leads to a cell-cycle progression from G1 to S phase. mTOR controls the activation of ribosomal protein translation and the initiation of cap-dependent translation. A role of mTOR signaling pathway dysregulation in tumourigenesis is postulated. mTOR and pathways upstream of this kinase were found to be frequently upregulated in neoplastic diseases. Therefore, it is also an attractive target for antitumour therapy. Several mTOR inhibitors were developed, including rapamycin and its analogues: CCI-779, RAD001 and AP23573. After promising phase I studies, their potential clinical significance is currently under evaluation in several phase II-III trials on patients with solid tumours and some hematological malignancies. PMID- 16217559 TI - Construction of a stable cell line uniformly expressing the rat TRPV1 receptor. AB - We constructed and analyzed a new cell line called HT5-1, which stably expresses an enhanced green fluorescent protein-tagged version of the rat vanilloid receptor 1 (VR1/TRPV1). The fluorescent receptor allowed easy measurement of receptor expression and expression level-based purification of cells via fluorescence-activated cell sorting. The HT5-1 cell line was compared to cells transiently transfected with the fluorescent receptor, to cells expressing the native rat vanilloid receptor, and to isolated capsaicin-sensitive rat trigeminal sensory neurons. Fura-2 microfluorimetry measurements of the calcium influx upon capsaicin induction showed that, by contrast to transiently transfected cells, HT5-1 cells respond uniformly to the stimulation, due to the similar level of receptor expression in individual cells. HT5-1 cells showed similar behaviour to isolated trigeminal root ganglion neurons, including marked tachyphylaxis upon repeated capsaicin induction, and a lack of calcium ion release from intracellular storage sites. PMID- 16217560 TI - The correlation between oxidative stress and leaf senescence during plant development. AB - In plants, besides being the final step leading to the death of the whole organism, senescence has a developmental function involving the coordinated degradation of macromolecules and the mobilization of nutrients out of senescing tissues into developing parts of the plant. Free radicals are thought to play an essential role in senescence, especially those derived from oxygen. Since these molecules are extremely toxic, the levels of the different reactive oxygen species have to be tightly regulated. However, at low concentrations, hydrogen peroxide may also serve as a signalling molecule. Therefore, a coordinated regulation of the free radical scavenging system, which comprises enzymatic components such as catalase, superoxide dismutase and ascorbate peroxidase, and non-enzymatic molecules such as ascorbate and glutathione is essential. The increased radical levels displayed during senescence are not only caused by the elevated production of radicals but also by a loss in antioxidant capacity. PMID- 16217562 TI - Tackling health inequalities--general practitioners and physicians working together. PMID- 16217561 TI - Rat colonic lipid peroxidation and antioxidant status: the effects of dietary luteolin on 1,2-dimethylhydrazine challenge. AB - Colon cancer is the third most common cancer and second leading cause of cancer related death in the United States. A number of recent articles demonstrate the importance of natural products as cancer chemopreventive agents. In this study, we evaluated the chemopreventive efficacy of luteolin, a flavonoid, on tissue lipid peroxidation and antioxidant status, which are used as biomarkers in DMH induced experimental colon carcinogenesis. Rats were given a weekly subcutaneous injection of DMH at a dose of 20 mg/kg body weight for 15 weeks. Luteolin (0.2 mg/kg body weight/everyday p.o.) was given to the DMH-treated rats at the initiation and post-initiation stages of carcinogenesis. The animals were killed after 30 weeks. After a total experimental period of 32 weeks (including 2 weeks of acclimatization), tumor incidence was 100% in DMH-treated rats. In those DMH treated rats that had received luteolin during the initiation or post-initiation stages of colon carcinogenesis, the incidence of cancer and the colon tumor size was significantly reduced as compared to that for DMH-treated rats not receiving luteolin. In the presence of DMH, relative to the results for the control rats, there were decreased levels of lipid peroxidation, as denoted by thiobarbituric acid reactive substances (TBARS), conjugated dienes and lipid hydroperoxides, decreased activities of the enzymic antioxidants superoxide dismutase (SOD) and catalase (CAT), and elevated levels of glutathione and the glutathione-dependent enzymes reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione-S transferase (GST) and glutathione reductase (GR), and of the non-enzymic antioxidants vitamin C and vitamin E. Our study shows that intragastric administration of luteolin inhibits colon carcinogenesis, not only by modulating lipid peroxidation and antioxidant status, but also by preventing DMH-induced histopathological changes. Our results thus indicate that luteolin could act as a potent chemopreventive agent for colon carcinogenesis. PMID- 16217563 TI - Encounters with indigenous patients in Australian general practice. AB - The BEACH program, a continuous national study of general practice activity in Australia, gives an overview of consultations with indigenous patients. Between 1998 and 2003, general practitioners recorded 5476 consultations with people who identified themselves as being of Aboriginal and/or Torres Strait Islander origin. These accounted for approximately 1% of total BEACH encounters for the 5 year period. In this article we compare 'indigenous encounters' with total BEACH encounters. This provides a backdrop against which articles in this issue of Australian Family Physician can be further considered. PMID- 16217564 TI - Indigenous male health disadvantage--linking the heart and mind. AB - BACKGROUND: Aboriginal men experience unacceptably high morbidity and mortality rates for almost all medical conditions. Heart disease and depression in particular are common, inter-linked, and potentially amendable to interventions delivered through primary care. OBJECTIVE: This article aims to raise awareness of mental illness and heart disease in Aboriginal men, to explore the links between these disease groups, and to encourage general practitioners to do all they can to help reduce morbidity and mortality, in particular through active secondary prevention. DISCUSSION: It would be simplistic to think that these problems can be adequately addressed merely through improved general practice services, but improved recognition of illness by GPs, improved accessibility and acceptability of health services, active follow up and management of depression and heart disease, and ongoing engagement with communities is likely to improve the current situation. PMID- 16217565 TI - Prevention and socioeconomic disadvantage. AB - BACKGROUND: Counselling in behavioural risk factors links chronic disease prevention and chronic disease care in the day-to-day work of general practice. This is particularly so in diabetes and cardiovascular disease. Each of these conditions is significantly more common in socioeconomically disadvantaged communities, suggesting that preventive activity may be particularly important for these groups; but what does that mean for general practitioners working with individual patients in their practice? OBJECTIVE: This article sets out some broad approaches to making sure that preventive activity in general practice reaches effectively those living in adverse socioeconomic circumstances. DISCUSSION: Rather than different preventive care, we require extra and targeted effort and a modified approach. We need to ensure that preventive care reaches those most in need and is implemented in a way that is sensitive to patient context. Collecting data on patient socioeconomic status is an important step in applying an 'equity lens' to our preventive care. A practice team approach is required to develop clear goals and address any gaps identified in preventive care. At a one-to-one level we need to allocate extra time to patients as well as reflect on our own attitudes and assumptions about social disadvantage and health. PMID- 16217566 TI - Refugee health. AB - BACKGROUND: Refugees and asylum seekers in Australia come from many countries. They present a significant challenge for general practice. OBJECTIVE: This article outlines the unique range of problems presented by refugee patients in general practice and some approaches to dealing with them. DISCUSSION: Refugees and asylum seekers come to Australia with a range of health problems related to their experience both overseas and in Australia. These include the physical and psychological sequelae of torture and trauma such as anxiety, depression or post traumatic stress disorder, infectious diseases such as tuberculosis, as well as chronic illness. These problems need to be addressed in general practice, as should preventive care, which is often overlooked. While those on permanent or temporary visas will have access to Medicare, those on bridging visas may not. PMID- 16217567 TI - Concordance--An alternative term to 'compliance' in the Aboriginal population. AB - BACKGROUND: 'Compliance' is an important issue for the health of the Aboriginal people. The word implies that patients are not following the doctor's 'rules'. Concordance may be a better concept to describe both doctor and patient working together in 'harmony and agreement'. OBJECTIVE: This article describes some of the parameters that affect consultations with Aboriginal patients and how they might be better addressed. DISCUSSION: The broader socio-political issues, the 'ethnocentrism' of the doctor, the health literacy of the patient, a more 'patient centred' model of the doctor-patient interaction, and the support of organisations are some of the variables that can be improved. It is important for general practitioners not to have a pessimistic attitude toward these issues, to identify ways in which the best possible results can be achieved, and to work hard to accomplish them. PMID- 16217568 TI - Inequities in Aboriginal health--access to the Asthma 3+ Visit Plan. AB - BACKGROUND: The prevalence of asthma is higher in Aboriginal and Torres Strait Islander adults than in non-Indigenous Australian adults in all age groups, and hospitalisation rates for acute asthma episodes are much higher, indicating suboptimal prevention. A 2001-2002 Australian Federal Budget initiative, the Asthma 3+ Plan, encouraged doctors to talk to their patients about asthma management over at least three visits and provide an asthma action plan. OBJECTIVE: This article reports on the findings of a study examining the uptake of the Asthma 3+ Visit Plan initiative among Aboriginal community controlled health services. DISCUSSION: Significant barriers prevent ACCHSs accessing the Asthma 3+ Visit Plan limiting Aboriginal people's capacity to benefit. In addition, Aboriginal and Torres Strait Islander patient access to both spacer devices and asthma medication is often poor. A targeted asthma information strategy is needed, and spacer devices must be made accessible to the Aboriginal and Torres Strait Islander population. Chronic disease management Medicare items offer a preferable and alternative funding mechanism for asthma care, if supplemented by pharmaceutical access reforms. PMID- 16217569 TI - PBS medications--improving access for Aboriginal and Torres Strait Islander peoples. AB - BACKGROUND: Expenditure through major commonwealth funded health programs such as the Pharmaceutical Benefits Scheme (PBS) is much lower for Aboriginal and Torres Strait Islander peoples than other Australians. Section 100 of the National Health Act (1953) allows for special access arrangements where pharmaceutical benefits cannot be conveniently supplied. OBJECTIVE: This article discusses the barriers to accessing PBS medications for Aboriginal and Torres Strait Islander peoples and the S100 access scheme. DISCUSSION: The implementation of S100 medications for remote area Aboriginal health services (AHSs) represents a breakthrough in medicines access, and is one of the most significant improvements in health service delivery for many years. If we are to achieve equity in access to the PBS for all Aboriginal and Torres Strait Islander peoples, an extension of this initiative is necessary for rural and urban AHSs. PMID- 16217570 TI - THE RACGP--supporting GPs to work better in Aboriginal and Torres Strait Islander health. AB - BACKGROUND: The Royal Australian College of General Practitioners (RACGP) has worked with funding from the Australian government to improve support and training for general practitioners working in Aboriginal and Torres Strait Islander communities. OBJECTIVE: This article outlines the role being played by the RACGP and addresses the question: 'What can I do?' for interested GPs. DISCUSSION: The RACGP plays an important role in supporting GPs working in Aboriginal and Torres Strait Islander communities. This includes current RACGP Aboriginal and Torres Strait Islander health projects such as providing support for the NACCHO GP Network, provision of accessible relevant cultural safety training for GPs, a national meeting of those working in Aboriginal and Torres Strait Islander health, access to the RACGP library's Aboriginal and Torres Strait Islander health resource collection, and the ongoing distribution of other resources, as well as RACGP organisational support for a range of other initiatives. On an individual level, all GPs are challenged to work at being well informed, and to take a personal leadership role so they can play a part in improving Aboriginal and Torres Strait Islander health. PMID- 16217571 TI - NACCHO GP Network--enhancing communication in Aboriginal health. AB - BACKGROUND: Communication between general practitioners, information sharing, and GP support are important issues, especially for those in rural and remote areas. Internet based links can facilitate enhanced communication. OBJECTIVE: This article describes the new internet based communication forum for GPs working, or interested in the health of, Aboriginal and Torres Strait Islander peoples. DISCUSSION: The NACCHO GP Network is the first on-line service to provide focussed information for GPs in the Aboriginal community controlled health sector. It also provides a place for GPs, locums and general practice registrars to raise issues and share ideas and information on Aboriginal health practice. The NACCHO GP Network has the potential to reduce isolation and address the support needs of GPs, and enhance the interaction of the Aboriginal community controlled sector with general practice organisations. PMID- 16217572 TI - Breast cancer in young women. AB - This eighth article in our series on breast disease focuses on breast cancer in young women. There are several challenges in the diagnosis of breast cancer in young women, and special consideration must be given to the woman's fertility and body image issues. The difficulty in distinguishing benign from malignant lesions on clinical and imaging assessment in this group stresses the importance of the triple test of clinical examination, mammography or ultrasound, and biopsy. PMID- 16217573 TI - Ear examination--a practical guide. PMID- 16217574 TI - Re-entry adjustment of cross cultural workers--the role of the GP. AB - BACKGROUND: Re-entry adjustment affects Australian cross cultural workers returning home; and for many, loss and grief issues arise. General practitioners are often the first point of contact in the health care system and are well placed to deal with these issues. OBJECTIVE: This article examines strategies that GPs can use to support the Australian cross cultural worker on re-entry, and focuses on recognition of re-entry adjustment, the role of loss and grief issues, and the importance of dealing with these issues. DISCUSSION: Australian cross cultural workers are valued members of their communities. However, their loss and grief issues associated with re-entry adjustment on return are often unrecognised and may lead to significant morbidity. Acknowledgment of their disenfranchised grief and appropriate therapy may be part of the role of their GP. Further research is needed to equip GPs to manage this important group in the Australian community. PMID- 16217575 TI - Diabetes and the skin. Part 1--tender skin lumps. PMID- 16217576 TI - Contraception and sexually transmitted infections. AB - BACKGROUND: Many women seek contraceptive advice from general practitioners. These consultations provide an opportunity to discuss the prevention of sexually transmitted infections (STIs) including human immunodeficiency virus (HIV). OBJECTIVE: This article discusses the effect of various contraceptive methods on the transmission of STIs and HIV. It also highlights issues relevant to achieving both contraception and STI/HIV prevention, with a focus on younger women. DISCUSSION: There is no ideal method to achieve protection against both pregnancy and STIs/HIV. Counselling about risks of STIs/HIV, providing the option of being tested for bacterial STIs, hepatitis B and HIV at the start of new relationships and promoting condom use for casual sex and concurrent sexual relationships is to be encouraged. PMID- 16217577 TI - Self directed learning and continuing medical education. AB - Self directed learning (SDL) for continuing medical education (CME) is the most effective approach for improving physician performance and patient care outcomes. Self directed learning is an essential basis for CME. However, instructor directed programs--particularly on-site conferences--remain the most popular method for acquiring CME. This article briefly examines the evolution of SDL and its importance in lifelong learning for general practice. The challenge for CME providers is to facilitate SDL while taking general practitioners' views and preferences into consideration. PMID- 16217578 TI - A GP's duty of confidentiality. AB - Confidentiality is an accepted and fundamental basis of the doctor-patient relationship. Patients have a right to expect that information held about them will be held in confidence by their general practitioner. This article examines the concept of doctor-patient confidentiality and outlines the exceptions to the duty of confidentiality. PMID- 16217579 TI - Do we need USPs? AB - Australia is lacking the vital data relating to current practices needed to perform a meaningful evaluation of the quality of care in general practice. This article proposes that unannounced standardised patients (USPs) represent a valuable method of measuring actual performance in general practice. Constructive debate about the use of USPs may progress its acceptance as a valid tool for performance assessment and quality improvement. PMID- 16217580 TI - Research and development in Central Australia--an indigenous perspective. PMID- 16217581 TI - Indigenous research--a personal perspective. PMID- 16217582 TI - Consent before testing men for prostate cancer--a challenge? PMID- 16217583 TI - An educational workshop on the early detection of prostate cancer--a before-after evaluation. PMID- 16217584 TI - Early management of meningococcal disease--do attitudes of GPs influence practice? AB - BACKGROUND: Survival from early meningococcal disease might be improved if general practitioners followed guidelines by immediately administering parenteral antibiotics (before hospital referral). METHODS: Structured telephone interviews with 20 GPs who had previously treated meningococcal cases. RESULTS: General practitioners knew guideline recommendations for early management of meningococcal disease: early parenteral antibiotics would be given by about half the GPs entertaining a diagnosis of meningococcal infection. Barriers to immediate treatment were: diagnostic uncertainty, regarding the case as nonurgent, and practising close to a hospital. DISCUSSION: Diagnosing meningococcal disease is difficult in general practice. Early antibiotic administration for suspected cases is appropriate even in close proximity to referral hospitals. PMID- 16217585 TI - What factors facilitate a GP survey high response rate? PMID- 16217586 TI - Evidence-based guidelines for the prevention and treatment of glucocorticoid induced osteoporosis: a consensus document of the Belgian Bone Club. AB - Glucocorticoids (GCs) are frequently prescribed for various inflammatory and/or life-threatening conditions concerning many systems in the body. However, they can provoke many aftereffects, of which osteoporosis (OP) is one of the most crippling complications, with its host of fractures. The dramatic increase in bone fragility is mainly attributable to the GC-induced rapid bone loss in all skeletal compartments. We have reviewed the meta-analyses and randomized controlled studies reporting medical therapeutic interventions currently registered in Belgium for the management of GC-OP comparatively with a placebo. Based on this research, an expert meeting developed a consensus on the prevention and therapy of GC-OP. The pathophysiology of GC-OP is complex. Several factors, acting separately or synergistically, have been described. Their great number could help to understand the rapidity of bone loss and of bone fragility occurrence, indicating that a rapid therapeutic intervention should be implemented to avoid complications. All patients on GCs are threatened with OP, so the prevention and/or therapy of GC-OP should be considered not only for postmenopausal females, but also for osteopenic premenopausal females and for males put on a daily dose of at least 7.5 mg equivalent prednisolone that is expected to last at least 3 months. Non-pharmacological interventions, such as exercise and avoidance of tobacco and alcohol, should be recommended, even if their role is not definitely settled in GC-OP prevention. Supplemental calcium and vitamin D should be considered as the first-line therapy because of the decrease in intestinal calcium absorption provoked by GCs. They also could be considered either as isolated therapy in patients taking less than 7.5 mg prednisolone daily and/or for a predicted period shorter than 3 months or as adjuvant therapy to other more potent drugs. Hormone replacement therapy could be considered in young postmenopausal females on GC, such as in postmenopausal OP, or in men with low androgen levels. Calcitonin appears to have a protective effect on trabecular bone in GC-OP, just as in postmenopausal OP. There is an increasing body of evidence supporting the antifracture efficacy of bisphosphonates, notably alendronate and risedronate. Preventative and curative therapy of GC-OP should be maintained as long as the patient is on GC treatment and could be stopped after weaning from GC, because there is more than circumstantial evidence of some recovery of BMD when GCs are stopped. There is no indication in GC-OP for any combination of two antiresorptive agents (except for calcium and vitamin D) or for an antiresorptive and an anabolic agent. There is indeed no proof that the increased costs of combined treatments will translate into increased therapeutic efficacy. PMID- 16217588 TI - Anti-vertebral fracture efficacy of raloxifene: a meta-analysis. AB - In the Multiple Outcomes of Raloxifene Evaluation (MORE) trial, raloxifene reduced the risk of vertebral fracture. However, a systematic analysis of the anti-vertebral fracture efficacy of raloxifene, which includes the results of newly reported studies, has not been performed. A meta-analysis was carried out using all randomized, double-blind, placebo-controlled trials to determine whether the reduction in the risk for vertebral fracture, reported with raloxifene, was consistent among studies, and to define more accurately the point estimate of the odds ratio. Three prevention studies, two arms of the MORE trial, and three additional treatment studies in which fracture data were available from prospectively scheduled spinal radiographs were included in the analysis. A systematic review of the literature (MedLine, EMBASE) confirmed that no studies with raloxifene had been excluded from this analysis. The effects of raloxifene 60 mg/day (RLX60) and 120 mg/day pooled with 150 mg/day (RLX120/150) were analyzed by intention to treat. There was no significant heterogeneity among the studies included in the meta-analysis. Odds ratio estimates (95% CI) were 0.60 (0.49, 0.74) for RLX60 and 0.51 (0.41, 0.64) for RLX120/150. From these data we infer that raloxifene consistently reduces the risk of vertebral fracture in postmenopausal women. PMID- 16217587 TI - Incidence and cost of osteoporotic fractures in France during 2001. A methodological approach by the national hospital database. AB - The objective of this study was to describe the hospital incidence rate and the in-patient costs of three peripheral "osteoporotic" fractures (proximal humerus and hip, distal radius and/or ulna) in women and men aged over 45 in France during 2001. Each stay for fracture was selected from the dataset of the French national hospital database in 2001. The incidence rate (CI 95%) was standardized by age and gender according to the last census of the French population (1999). The effect of age and existence of geographical difference in incidence rates has been studied. For each fracture, we described the number of stays, rate of surgical procedure and in-patient costs according to the 2004 French list of cost per diagnosis-related group (2004 Euros); 118,839 fractures were registered during 2001 (61% hip, 28% distal radius and 11% proximal humerus; sex ratio 0.26). The incidence rate for all fracture was 7,567 (7,519-7,615) and 2,312 (2,283-2,341) for 10(6) inhabitants in women and men aged over 45 years, respectively. The incidence increased significantly whatever type of fracture and gender. There were more fracture incidents in the east of France compared to the west and in the south compared to the north, whatever type of fracture in women and only for hip fracture in men. Surgical procedures were performed in 91% of proximal hip fractures, 83% of distal radius fractures and 53% of proximal humerus fractures. The median in-patient costs were 3,786 Euros for the humerus, from 2,363 to 2,574 Euros for the radius and from 8,048 to 8,727 Euros for the hip. The evaluation of the burden of peripheral fractures is possible using national hospital data in France. The incidence of fractures increased with age and is more common in women. Hip fracture with its higher occurrence, rate of procedure and in-patient costs could be used as a marker of osteoporosis for evaluating strategies of management. PMID- 16217591 TI - Association analysis of interleukin 5 receptor alpha subunit (IL5RA) polymorphisms and asthma. AB - The alpha subunit of interleukin 5 receptor (IL5RA) on chromosome 3p26-p24 is known to regulate the development and function of eosinophils. In an effort to discover additional polymorphism(s) in genes whose variant(s) have been implicated in asthma, we investigated the genetic polymorphisms in IL5RA to evaluate the gene as a potential candidate for a host genetic study of asthma. By direct DNA sequencing in 24 individuals, we identified 22 sequence variants within exons and flanking regions including a 1.5-kb promoter region of IL5RA; 10 common polymorphic sites were selected for genotyping in our asthma cohort (n = 587). Two haplotype blocks were identified in a Korean population. Statistical analysis revealed that one promoter SNP, c.-5993A > G, and one ins/del polymorphism in intron 3, c.-480_482insdelGTT, showed significant association with the risk of asthma development. The genetic effects of c.-5993A>G and c. 480_482insdelGTT on asthma were more apparent among atopic subjects. Our findings suggest that polymorphisms in IL5RA might be among the genetic risk factors for asthma development, especially in atopic populations. IL5RA variant/haplotype information identified in this study will provide valuable information for strategies for the control of asthma. PMID- 16217589 TI - Osteoporosis after spinal cord injury. AB - Osteoporosis is a known consequence of spinal cord injury (SCI) and occurs in almost every SCI patient. It manifests itself as an increase in the incidence of lower extremity fractures. The pattern of bone loss seen in SCI patients is different from that usually encountered with endocrine disorders and disuse osteoporosis. In general, there is no demineralization in supralesional areas following SCI. Several factors appear to have a major influence on bone mass in SCI individuals, such as the degree of the injury, muscle spasticity, age, sex and duration after injury. At the lumbar spine, bone demineralization remains relatively low compared to that of the long bones in the sublesional area. A new steady state level between bone resorption and formation is reestablished about 2 years after SCI. SCI may not only cause bone loss, but also alter bone structure and microstructure. Trabecular bone is more affected than cortical bone in the SCI population. Numerous clinical series have reported a high incidence ranging from 1 to 34% of lower extremity fractures in SCI patients. The pathogenesis of osteoporosis after SCI remains complex and perplexing. Disuse may play an important role in the pathogenesis of osteoporosis, but neural factors also appear to be important. SCI also leads to impaired calcium and phosphate metabolism and the parathyroid hormone (PTH)-vitamin D axis. Pharmacologic intervention for osteoporosis after SCI includes calcium, phosphate, vitamin D, calcitonin and biphosphonates. However, the concomitant prescription of bone active drugs for the prevention and treatment of osteoporosis remains low, despite the availability of effective therapies. Functional stimulated exercises may contribute to the prevention of bone loss to some extent. In addition, many unanswered questions remain about the pathogenesis of osteoporosis and its clinical management. PMID- 16217590 TI - Survival after hip fracture. AB - Although it is known that overall mortality is increased after hip fracture, the influence of hip fracture risk factors on the subsequent mortality and cause of death has not been well studied. The objective of this study was to establish the survival after hip fracture in women and to assess the impact of comorbidity on mortality. We identified a complete population-based set of 2,245 incident hip fracture cases and 4,035 randomly selected population-based controls among women 50-81 years old in Sweden and followed these subjects for an average of 5 years through the Swedish National Inpatient and Cause-of-Death Registers. Information on factors related to hip fracture was obtained through linkage to hospital discharge data and through a mailed questionnaire. We studied excess mortality of hip fracture patients compared to controls using survival curves and proportional hazard regression models. During follow-up, 896 hip fracture patients (40%) and 516 (13%) controls died. The relative risk (RR) of death, adjusted for age and previous hospitalization for serious disease, was 2.3 (95% CI 2.0-2.5). Although the highest mortality risks were in the 1st 6 months post-fracture, RRs for fractures versus controls were increased for at least 6 years. Increased mortality was apparent both in those with evidence of comorbidity and those without. Hip fracture patients have a substantially increased risk of death that persists for at least 6 years post-fracture. The relative excess mortality is independent of comorbidity and known hip fracture risk factors. PMID- 16217592 TI - Aortic valve insufficiency in patients with chronic rheumatic diseases. AB - Aortic valve lesions are often found in patients with rheumatic diseases, but their clinical significance has not been properly evaluated. In the present study, the echocardiographic files of the cardiology unit of the Oulu University Hospital were screened for a diagnosis of aortic insufficiency (AI). The aetiology of the valve disease and specific details of the rheumatic disease were evaluated in 160 patients. Twenty-eight patients (18%) had a history of rheumatic fever. Rheumatic disease was found in 14 patients (8.8%) with AI, which is significantly more than the prevalence of rheumatic diseases (1.8%) in the corresponding age group (35-100 years) in Finland. Rheumatoid arthritis or juvenile rheumatoid arthritis was found in seven patients (4.4%), whereas ankylosing spondylitis or seronegative spondylarthropathy were found in four patients (2.5%). Other rheumatic diseases included Takayasu's arteritis (two patients) and scleroderma (one patient). When 38 patients with pure AI without other possible aetiology were analysed, rheumatic disease was found in five patients (13%). Patients with rheumatic disease as a potential aetiology of AI often had symptomatic valve disease, which required surgical treatment, although great differences between different aetiologies were not found. PMID- 16217593 TI - Public opinions and beliefs about the treatment of depression in urban Turkey. AB - BACKGROUND: Although attitudes towards psychiatric illness influence its presentation, detection, recognition, treatment adherence and rehabilitation, the lay public's opinions and beliefs about the treatment of depression have not been investigated sufficiently. OBJECTIVE: The aim of this study was to determine public opinions and beliefs about the treatment of depression and the influence of perception and causal attributions on attitudes towards treatment of depression in urban areas. METHODS: This study was carried out with a representative sample in Istanbul, which is the biggest metropolis in Turkey. Seven hundred and seven subjects completed the public survey form which consisted of 32 items rating attitudes towards depression. RESULTS: The public believes that psychological and social interventions are more effective than pharmacotherapy, and that the medicines used in treatment of depression are harmful and addictive. There was a general reluctance to consult a physician for depression, and psychiatrists were felt to be more helpful than general practitioners. The public viewed depression as treatable. A high educational level and perceiving depression as a disease is associated with positive beliefs and opinions about the treatment of depression; but the perception of depressive patients as aggressive is associated with negative beliefs and opinions about the treatment of depression. CONCLUSION: The beliefs that "psychological and social interventions are more effective than pharmacotherapy" and "antidepressants are harmful and addictive" must specifically be taken into account in clinical practice and in anti-stigma campaigns. Additional studies are needed to understand the public's tendency to conceptualise depression as a psychosocial problem. In clinical practice, depression should be introduced as a bio psychosocial disease whatever its cause: biological, psychological or social. In addition, the differences between extreme worry and disease, and the lack of aggressiveness of depressive patients, must be emphasised. PMID- 16217594 TI - Suicide among adolescents. A psychological autopsy study of psychiatric, psychosocial and personality-related risk factors. AB - BACKGROUND: The suicide rate among young males in Belgium has doubled over the last decade. As more knowledge about risk factors is required to develop national prevention strategies, we investigated adolescent suicides using the psychological autopsy method. METHODS: A total of 32 informants were interviewed regarding 19 suicide cases (aged 15-19). A semi-structured interview schedule, constructed by Houston et al. (J Affect Disord 63:159-170, 2001), was used. RESULTS: All adolescents were suffering from one or more mental disorder(s) at the time of their death, and almost half of them were diagnosed with personality disorders. Adjustment disorders were diagnosed in one fifth of the sample, which appears to be relevant in view of the multiple life events and other psychosocial problems which adolescents were facing shortly before death. This suggests that difficulties in coping with stressful psychosocial problems are important in the course of the suicidal process. Only a small minority was receiving treatment for their disorders. CONCLUSIONS: Mental disorders, commonly untreated and combined with personality disorders and psychosocial problems, are frequently found in young suicide victims. This study suggests that education in the diagnosis and treatment of depression, adjustment disorders and suicide is important in the prevention of suicide. PMID- 16217597 TI - What it will take to feed 5.0 billion rice consumers in 2030. AB - Major advances have occurred in rice production due to adoption of green revolution technology. Between 1966 and 2000, the population of densely populated low income countries grew by 90% but rice production increased by 130% from 257 million tons in 1966 to 600 million tons in 2000. However, the population of rice consuming countries continues to grow and it is estimated that we will have to produce 40 more rice in 2030. This increased demand will have to be met from less land, with less water, less labor and fewer chemicals. To meet the challenge of producing more rice from suitable lands we need rice varieties with higher yield potential and greater yield stability. Various strategies for increasing the rice yield potential being employed include: (1) conventional hybridization and selection procedures, (2) ideotype breeding, (3) hybrid breeding, (4) wide hybridization and (5) genetic engineering. Various conventional and biotechnology approach are being employed to develop durable resistance to diseases and insect and for tolerance to abiotic stresses. The availability of the rice genome sequence will now permit identification of the function of each of 60,000 rice genes through functional genomics. Once the function of a gene is identified, it will be possible to develop new rice varieties by introduction of the gene through traditional breeding in combination with marker aided selection or direct engineering of genes into rice varieties. PMID- 16217595 TI - Expressed emotion and attributions in relatives of schizophrenia patients with and without substance misuse. AB - OBJECTIVE: To test the hypotheses that carers of patients with schizophrenia (single diagnosis) and schizophrenia and co-occurring drug or alcohol misuse (dual diagnosis) will differ in terms of expressed emotion (EE) and their attributions for patient problems. METHOD: In a cross-sectional study, two samples of 42 single- and dual-diagnosis carers are compared in terms of EE and attributions. Patient symptoms are assessed to control for differences other than substance misuse. RESULTS: The study supports the hypothesis that high-EE, dual diagnosis carers tend to see patient problems as more blaming (internal, controllable and personal) than do single-diagnosis patients. This difference was particularly marked when making causal ascriptions for deficit behaviours. Although there were no differences in overall EE levels in the two groups, there were significantly more carers who were rated as hostile and rejecting in the dual-diagnosis group. CONCLUSIONS: The findings highlight the importance of family intervention for this patient group. PMID- 16217599 TI - Where does the accurate rice genome sequence lead us? PMID- 16217598 TI - How can we use genomics to improve cereals with rice as a reference genome? AB - Rice serves as a model crop for cereal genomics. The availability of complete genome sequences, together with various genomic resources available for both rice and Arabidopsis, have revolutionized our understanding of the genetic make-up of crop plants. Both macrocolinearity revealed by comparative mapping and microcolinearity revealed by sequence comparisons among the grasses indicate that sequencing and functional analysis of the rice genome will have a significant impact on other cereals in terms of both genomic studies and crop improvement. The availability of mutants, introgression libraries, and advanced transformation techniques make functional genomics in rice and other cereals more manageable than ever before. A wide array of genetic markers, including anchor markers for comparative mapping, SSRs and SNPs are widely used in genetic mapping, germplasm evaluation and marker assisted selection. An integrated database that combines genome information for rice and other cereals is key to the effective utilization of all genomics resources for cereal improvement. To maximize the potential of genomics for plant breeding, experiments must be further miniaturized and costs must be reduced. Many techniques, including targeted gene disruption or allele substitution, insertional mutagenesis, RNA interference and homologous recombination, need to be refined before they can be widely used in functional genomic analysis and plant breeding. PMID- 16217600 TI - Genome-wide introgression lines and their use in genetic and molecular dissection of complex phenotypes in rice (Oryza sativa L.). AB - Tremendous efforts have been taken worldwide to develop genome-wide genetic stocks for rice functional genomic (FG) research since the rice genome was completely sequenced. To facilitate FG research of complex polygenic phenotypes in rice, we report the development of over 20,000 introgression lines (ILs) in three elite rice genetic backgrounds for a wide range of complex traits, including resistances/tolerances to many biotic and abiotic stresses, morpho agronomic traits, physiological traits, etc., by selective introgression. ILs within each genetic background are phenotypically similar to their recurrent parent but each carries one or a few traits introgressed from a known donor. Together, these ILs contain a significant portion of loci affecting the selected complex phenotypes at which allelic diversity exists in the primary gene pool of rice. A forward genetics strategy was proposed and demonstrated with examples on how to use these ILs for large-scale FG research. Complementary to the genome wide insertional mutants, these ILs opens a new way for highly efficient discovery, candidate gene identification and cloning of important QTLs for specific phenotypes based on convergent evidence from QTL position, expression profiling, functional and molecular diversity analyses of candidate genes, highlights the importance of genetic networks underlying complex phenotypes in rice that may ultimately lead to more complete understanding of the genetic and molecular bases of quantitative trait variation in rice. PMID- 16217601 TI - The oryza map alignment project: the golden path to unlocking the genetic potential of wild rice species. AB - The wild species of the genus Oryza offer enormous potential to make a significant impact on agricultural productivity of the cultivated rice species Oryza sativa and Oryza glaberrima. To unlock the genetic potential of wild rice we have initiated a project entitled the 'Oryza Map Alignment Project' (OMAP) with the ultimate goal of constructing and aligning BAC/STC based physical maps of 11 wild and one cultivated rice species to the International Rice Genome Sequencing Project's finished reference genome--O. sativa ssp. japonica c. v. Nipponbare. The 11 wild rice species comprise nine different genome types and include six diploid genomes (AA, BB, CC, EE, FF and GG) and four tetrapliod genomes (BBCC, CCDD, HHKK and HHJJ) with broad geographical distribution and ecological adaptation. In this paper we describe our strategy to construct robust physical maps of all 12 rice species with an emphasis on the AA diploid O. nivara -thought to be the progenitor of modern cultivated rice. PMID- 16217602 TI - Molecular evidence for asymmetric evolution of sister duplicated blocks after cereal polyploidy. AB - Polyploidy (genome duplication) is thought to have contributed to the evolution of the eukaryotic genome, but complex genome structures and massive gene loss during evolution has complicated detection of these ancestral duplication events. The major factors determining the fate of duplicated genes are currently unclear, as are the processes by which duplicated genes evolve after polyploidy. Fine scale analysis between homologous regions may allow us to better understand post polyploidy evolution. Here, using gene-by-gene and gene-by-genome strategies, we identified the S5 region and four homologous regions within the japonica genome. Additional phylogenomic analyses of the comparable duplicated blocks indicate that four successive duplication events gave rise to these five regions, allowing us to propose a model for this local chromosomal evolution. According to this model, gene loss may play a major role in post-duplication genetic evolution at the segmental level. Moreover, we found molecular evidence that one of the sister duplicated blocks experienced more gene loss and a more rapid evolution subsequent to two recent duplication events. Given that these two recent duplication events were likely involved in polyploidy, this asymmetric evolution (gene loss and gene divergence) may be one possible mechanism accounting for the diploidization at the segmental level. PMID- 16217603 TI - The plant architecture of rice (Oryza sativa). AB - Plant architecture, a collection of the important agronomic traits that determine grain production in rice, is mainly affected by factors including tillering, plant height and panicle morphology. Recently, significant progress has been made in isolating and collecting of mutants that are defective in rice plant architecture. Although our understanding of the molecular mechanisms that control rice tillering, panicle development and plant height are still limited, new findings have begun to emerge. This review, therefore, summarizes the recent progress in exploring the mechanisms that control rice plant architecture. PMID- 16217604 TI - Chemical- and irradiation-induced mutants of indica rice IR64 for forward and reverse genetics. AB - IR64, the most widely grown indica rice in South and Southeast Asia, possesses many positive agronomic characteristics (e.g., wide adaptability, high yield potential, tolerance to multiple diseases and pests, and good eating quality,) that make it an ideal genotype for identifying mutational changes in traits of agronomic importance. We have produced a large collection of chemical and irradiation-induced IR64 mutants with different genetic lesions that are amenable to both forward and reverse genetics. About 60,000 IR64 mutants have been generated by mutagenesis using chemicals (diepoxybutane and ethylmethanesulfonate) and irradiation (fast neutron and gamma ray). More than 38,000 independent lines have been advanced to M4 generation enabling evaluation of quantitative traits by replicated trials. Morphological variations at vegetative and reproductive stages, including plant architecture, growth habit, pigmentation and various physiological characters, are commonly observed in the four mutagenized populations. Conditional mutants such as gain or loss of resistance to blast, bacterial blight, and tungro disease have been identified at frequencies ranging from 0.01% to 0.1%. Results from pilot experiments indicate that the mutant collections are suitable for reverse genetics through PCR detection of deletions and TILLING. Furthermore, deletions can be detected using oligomer chips suggesting a general technique to pinpoint deletions when genome wide oligomer chips are broadly available. M4 mutant seeds are available for users for screening of altered response to multiple stresses. So far, more than 15,000 mutant lines have been distributed. To facilitate broad usage of the mutants, a mutant database has been constructed in the International Rice Information System (IRIS; http: //www.iris.irri.org) to document the phenotypes and gene function discovered by users. PMID- 16217605 TI - EU-OSTID: a collection of transposon insertional mutants for functional genomics in rice. AB - A collection of 1373 unique flanking sequence tags (FSTs), generated from Ac/Ds and Ac transposon lines for reverse genetics studies, were produced in japonica and indica rice, respectively. The Ds and Ac FSTs together with the original T DNAs were assigned a position in the rice genome sequence represented as assembled pseudomolecules, and found to be distributed evenly over the entire rice genome with a distinct bias for predicted gene-rich regions. The bias of the Ds and Ac transposon inserts for genes was exemplified by the presence of 59% of the inserts in genes annotated on the rice chromosomes and 41% present in genes transcribed as disclosed by their homology to cDNA clones. In a screen for inserts in a set of 75 well annotated transcription factors, including homeobox containing genes, we found six Ac/Ds inserts. This high frequency of Ds and Ac inserts in genes suggests that saturated knockout mutagenesis in rice using this strategy will be efficient and possible with a lower number of inserts than expected. These FSTs and the corresponding plant lines are publicly available through OrygenesDB database and from the EU consortium members. PMID- 16217607 TI - Conservation of the E-function for floral organ identity in rice revealed by the analysis of tissue culture-induced loss-of-function mutants of the OsMADS1 gene. AB - Rapid progress in studies on flower development has resulted in refining the classical 'ABC model' into a new 'ABCDE model' to explain properly the regulation of floral organ identity. Conservation of E-function for flower organ identity among the dicotyledonous (dicot) plants has been revealed. However, its conservation in monocotyledonous (monocot) plants remains largely unknown. Here, we show the conservation of E-function in rice (Oryza sativaL.) by characterizing tissue culture-induced mutants of two MADS-box genes, OsMADS1and OsMADS5, which form a subclade within the well-supported clade of SEP-genes (E-function) phylogeny. Severe loss-of-function mutations of OsMADS1cause complete homeotic conversion of organs (lodicules, stamens, and carpels) of three inner whorls into lemma- and palea-like structures. Such basic deformed structure is reiterated along with the pedicel at the center of the same floret, indicating the loss of determinacy of the flower meristem. These phenotypes resemble the phenotypes caused by mutations of the dicot E-class genes, such as the Arabidopsis SEP123(SEPALLATA1/2/3) and the petunia FBP2(Floral Binding Protein 2), suggesting that OsMADS1play a very similar role in rice to that of defined E-class genes in dicot plants. In case of the loss-of-function mutation of OsMADS5, no defect in either panicles or vegetative organs was observed. These results demonstrate that OsMADS1clearly possesses E-function, and so, E-function is fundamentally conserved between dicot plants and rice, a monocot model plant. PMID- 16217606 TI - Reverse genetic approaches for functional genomics of rice. AB - T-DNA and transposable elements e.g., Ds and Tos17, are used to generate a large number of insertional mutant lines in rice. Some carry the GUS or GFP reporter for gene trap or enhancer trap. These reporter systems are valuable for identifying tissue- or organ-preferential genes. Activation tagging lines have also been generated for screening mutants and isolating mutagenized genes. To utilize these resources more efficiently, tagged lines have been produced for reverse genetic approaches. DNA pools of the T-DNA tagged lines and Tos17 lines have been prepared for PCR screening of insertional mutants in a given gene. Tag end sequences (TES) of the inserts have also been produced. TES databases are beneficial for analyzing the function of a large number of rice genes. PMID- 16217608 TI - A pilot study of transcription unit analysis in rice using oligonucleotide tiling path microarray. AB - As the international efforts to sequence the rice genome are completed, an immediate challenge and opportunity is to comprehensively and accurately define all transcription units in the rice genome. Here we describe a strategy of using high-density oligonucleotide tiling-path microarrays to map transcription of the japonica rice genome. In a pilot experiment to test this approach, one array representing the reverse strand of the last 11.2 Mb sequence of chromosome 10 was analyzed in detail based on a mathematical model developed in this study. Analysis of the array data detected 77% of the reference gene models in a mixture of four RNA populations. Moreover, significant transcriptional activities were found in many of the previously annotated intergenic regions. These preliminary results demonstrate the utility of genome tiling microarrays in evaluating annotated rice gene models and in identifying novel transcription units that will facilitate rice genome annotation. PMID- 16217609 TI - Microarray analysis reveals similarities and variations in genetic programs controlling pollination/fertilization and stress responses in rice (Oryza sativa L.). AB - Previously, we identified 253 cDNAs that are regulated by pollination/fertilization in rice by using a 10K cDNA microarray. In addition, many of them also appeared to be involved in drought and wounding responses. To investigate this relationship, we obtained their expression profiles after dehydration and wounding treatments in this study. Venn diagram analysis indicated that 53.8% (136/253) and 21% (57/253) of the pollination/fertilization related genes are indeed regulated by dehydration and wounding, respectively, and nearly half of the genes expressed preferentially in unpollinated pistils (UP) are responsive to dehydration. These results indicated that an extensive gene set is shared among these responses, suggesting that the genetic programs regulating them are likely related. Among them, the genetic network of water stress control may be a key player in pollination and fertilization. Additionally, 39.5% (100/253) cDNAs that are related to pollination/fertilization appear not to be regulated by the stress treatments (dehydration and wounding), suggesting that the existence of additional genetic networks are involved in pollination/fertilization. Furthermore, comparative analysis of the expression profiles of the 253 cDNAs under 18 different conditions (various tissues, treatments and developmental status) revealed that the genetic networks regulating photosynthesis, starch metabolisms, GA- and defense-responses are involved in pollination and fertilization. Taken together, these results provided some clues to elucidate the molecular mechanisms of pollination and fertilization in rice. PMID- 16217610 TI - Profiling the expression of genes controlling rice grain quality. AB - Rice provides a staple source of energy, protein and other nutrients to half of the world population. Over 90 of the rice seeds consists of starch and protein by dry weight. The quantity and property of starch and protein thus play a dominant role in the yield and quality of rice. The amylase content of starch is a determining factor in the eating and cooking quality while the amount and essential amino acids balance of storage proteins affect the nutritional quality of rice. In China, the super-hybrid rice currently under the last phase of development has a 35 yield advantage over the best inbred rice varieties. However, its grain quality needs further improvement. This study reported the expression patterns of 44 genes participating in starch, storage protein, and lysine synthesis in the developing rice grain. Field grown rice cultivar 9311, the paternal line of an elite super-hybrid rice LYP9 with its draft genomic sequence released, was used as plant material. Results revealed diverse yet coordinated expression profiles of the genes involved in the three pathways which lead to the final composition and property of starch, protein and lysine that determine the quality of rice, providing useful information for rice quality improvement. PMID- 16217611 TI - Rice proteome database: a step toward functional analysis of the rice genome. AB - The technique of proteome analysis using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) has the power to monitor global changes that occur in the protein complement of tissues and subcellular compartments. In this study, the proteins of rice were cataloged, a rice proteome database was constructed, and a functional characterization of some of the identified proteins was undertaken. Proteins extracted from various tissues and subcellular compartments in rice were separated by 2D-PAGE and an image analyzer was used to construct a display of the proteins. The Rice Proteome Database contains 23 reference maps based on 2D-PAGE of proteins from various rice tissues and subcellular compartments. These reference maps comprise 13129 identified proteins, and the amino acid sequences of 5092 proteins are entered in the database. Major proteins involved in growth or stress responses were identified using the proteome approach. Some of these proteins, including a beta-tubulin, calreticulin, and ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice, have unexpected functions. The information obtained from the Rice Proteome Database will aid in cloning the genes for and predicting the function of unknown proteins. PMID- 16217612 TI - Transcription factors in rice: a genome-wide comparative analysis between monocots and eudicots. AB - It is not known how representative the Arabidopsis thaliana complement of transcription factors (TFs) is of other plants. The availability of rice (Oryza sativa) genome sequences makes possible a comparative analysis of TFs between monocots and eudicots, the two major monophyletic groups of angiosperms. Here, we identified 1611 TF genes that belong to 37 gene families in rice, comparable to the 1510 in Arabidopsis. Several gene subfamilies, but no families, were found to be lineage-specific. Phylogenetic analyses indicated that nearly half of the TF genes form clear orthologous pairs or groups, which were derived from 383 ancestral genes in the common ancestor of rice and Arabidopsis. Investigating gene duplication mechanisms revealed twelve pairs of large intragenomic duplicated blocks, which account for more than 40% of the rice genome. About 60% of the duplicated TF genes have been retained on duplicated segments. Functional conservation and diversification of TFs across monocot and eudicot lineages are discussed. PMID- 16217614 TI - The chemical senses, olfaction and taste: a tribute to Al Farbman. PMID- 16217615 TI - Personal reflections on 40 years of research in the chemical senses. PMID- 16217613 TI - Modification of endogenous natural genes by gene targeting in rice and other higher plants. AB - The capability to modify a genomic sequence into a designed sequence is a powerful tool for biologists and breeders to elucidate the function of an individual gene and its cis-acting elements of multigene families in the genome. Gene targeting refers to the alteration of a specific DNA sequence in an endogenous gene at its original locus in the genome. In higher plants, however, the overwhelming occurrence of the random integration of transgenes by non homologous end-joining is the main obstacle to develop efficient gene targeting. Two approaches have been undertaken to modify a genomic sequence in higher plants chimeric RNA/DNA oligonucleotide-directed gene targeting to generate a site specific base conversion, and homologous recombination-dependent gene targeting to produce either a base change or a gene replacement in a sequence-specific manner. The successful and reproducible targeting of an endogenous gene by homologous recombination, independently of gene-specific selection by employing a strong positive-negative selection, has been demonstrated for the first time in rice, an important staple food and a model plant for other cereal species. This review addresses the current status of targeting of an endogenous natural gene in rice and other higher plants and discusses possible models for Agrobacterium- mediated gene targeting by homologous recombination using a strong positive negative selection. PMID- 16217616 TI - Distinct roles for Sema3A, Sema3F, and an unidentified trophic factor in controlling the advance of geniculate axons to gustatory lingual epithelium. AB - Geniculate ganglion axons arrive in the lingual mesenchyme on embryonic day 13 (E13), 3-4 days before penetrating fungiform papilla epithelium (E17). This latency may result from chemorepulsion by epithelial Sema3A (Dillon et al. (2004) Journal of Comparative Neurology 470, 13-24), or Sema3F, which we report is also expressed in this epithelium. Sema3A and Sema3F repelled or suppressed geniculate neurite outgrowth, respectively, and these effects were stage and neurotrophic factor dependent. BDNF-stimulated outgrowth is repelled by Sema3A until E17, but insensitive to Sema3F from E16. NT-4-stimulated neurite outgrowth is sensitive to Sema3A and Sema3F through E18, but NT-4 has not been detected in E15-18 tongue. E15-18 tongue explants did not exhibit net chemorepulsion of geniculate neurites, but the ability of tongue explants to support geniculate neurite outgrowth fluctuates: E12-13 (Rochlin et al. (2000), Journal of Comparative Neurology, 422, 579-593) and E17-18 explants promote and may attract geniculate neurites, but stages corresponding to intralingual arborization do not. The E18 trophic and tropic effects were evident even in the presence of BDNF or NT-4, suggesting that some other factor is responsible. Intrinsic neurite outgrowth capability (without exogenous neurotrophic factors) fluctuated similarly: ganglia deteriorated at E15, but exhibited moderate outgrowth at E18. The chemorepulsion studies are consistent with a role for Sema3A, not Sema3F, in restricting geniculate axons from the epithelium until E17, when axons penetrate the epithelium. The transient inability of tongue explants to promote geniculate neurite outgrowth may signify an alternative mechanism for restricting geniculate axons from the epithelium: limiting trophic factor access. PMID- 16217617 TI - Lingual deficits in neurotrophin double knockout mice. AB - Brain-derived neurotrophic factor (BDNF) and Neurotrophin 3 (NT-3) are members of the neurotrophin family and are expressed in the developing and adult tongue papillae. BDNF null-mutated mice exhibit specific impairments related to innervation and development of the gustatory system while NT-3 null mice have deficits in their lingual somatosensory innervation. To further evaluate the functional specificity of these neurotrophins in the peripheral gustatory system, we generated double BDNF/NT-3 knockout mice and compared the phenotype to BDNF(-/ ) and wild-type mice. Taste papillae morphology was severely distorted in BDNF(-/ ) xNT-3(-/-) mice compared to single BDNF(-/-) and wild-type mice. The deficits were found throughout the tongue and all gustatory papillae. There was a significant loss of fungiform papillae and the papillae were smaller in size compared to BDNF(-/-) and wild-type mice. Circumvallate papillae in the double knockouts were smaller and did not contain any intraepithelial nerve fibers. BDNF(-/-) xNT-3(-/-) mice exhibited additive losses in both somatosensory and gustatory innervation indicating that BDNF and NT-3 exert specific roles in the innervation of the tongue. However, the additional loss of fungiform papillae and taste buds in BDNF(-/-) xNT-3(-/-) mice compared to single BDNF knockout mice indicate a synergistic functional role for both BDNF-dependent gustatory and NT-3 dependent somatosensory innervations in taste bud and taste papillae innervation and development. PMID- 16217618 TI - Taste placodes are primary targets of geniculate but not trigeminal sensory axons in mouse developing tongue. AB - Tongue embryonic taste buds begin to differentiate before the onset of gustatory papilla formation in murine. In light of this previous finding, we sought to reexamine the developing sensory innervation as it extends toward the lingual epithelium between E 11.5 and 14.5. Nerve tracings with fluorescent lipophilic dyes followed by confocal microscope examination were used to study the terminal branching of chorda tympani and lingual nerves. At E11.5, we confirmed that the chorda tympani nerve provided for most of the nerve branching in the tongue swellings. At E12.5, we show that the lingual nerve contribution to the overall innervation of the lingual swellings increased to the extent that its ramifications matched those of the chorda tympani nerve. At E13.0, the chorda tympani nerve terminal arborizations appeared more complex than those of the lingual nerve. While the chorda tympani nerve terminal branching appeared close to the lingual epithelium that of the trigeminal nerve remained rather confined to the subepithelial mesenchymal tissue. At E13.5, chorda tympani nerve terminals projected specifically to an ordered set of loci on the tongue dorsum corresponding to the epithelial placodes. In contrast, the lingual nerve terminals remained subepithelial with no branches directed towards the placodes. At E14.5, chorda tympani nerve filopodia first entered the apical epithelium of the developing fungiform papilla. The results suggest that there may be no significant delay between the differentiation of embryonic taste buds and their initial innervation. PMID- 16217619 TI - Building sensory receptors on the tongue. AB - Neurotrophins, neurotrophin receptors and sensory neurons are required for the development of lingual sense organs. For example, neurotrophin 3 sustains lingual somatosensory neurons. In the traditional view, sensory axons will terminate where neurotrophin expression is most pronounced. Yet, lingual somatosensory axons characteristically terminate in each filiform papilla and in each somatosensory prominence within a cluster of cells expressing the p75 neurotrophin receptor (p75NTR), rather than terminating among the adjacent cells that secrete neurotrophin 3. The p75NTR on special specialized clusters of epithelial cells may promote axonal arborization in vivo since its over expression by fibroblasts enhances neurite outgrowth from overlying somatosensory neurons in vitro. Two classical observations have implicated gustatory neurons in the development and maintenance of mammalian taste buds--the early arrival times of embryonic innervation and the loss of taste buds after their denervation in adults. In the modern era more than a dozen experimental studies have used early denervation or neurotrophin gene mutations to evaluate mammalian gustatory organ development. Necessary for taste organ development, brain-derived neurotrophic factor sustains developing gustatory neurons. The cardinal conclusion is readily summarized: taste buds in the palate and tongue are induced by innervation. Taste buds are unstable: the death and birth of taste receptor cells relentlessly remodels synaptic connections. As receptor cells turn over, the sensory code for taste quality is probably stabilized by selective synapse formation between each type of gustatory axon and its matching taste receptor cell. We anticipate important new discoveries of molecular interactions among the epithelium, the underlying mesenchyme and gustatory innervation that build the gustatory papillae, their specialized epithelial cells, and the resulting taste buds. PMID- 16217620 TI - Are there efferent synapses in fish taste buds? AB - In fish, nerve fibers of taste buds are organized within the bud's nerve fiber plexus. It is located between the sensory epithelium consisting of light and dark elongated cells and the basal cells. It comprises the basal parts and processes of light and dark cells that intermingle with nerve fibers, which are the dendritic endings of the taste sensory neurons belonging to the cranial nerves VII, IX or X. Most of the synapses at the plexus are afferent; they have synaptic vesicles on the light (or dark) cells side, which is presynaptic. In contrast, the presumed efferent synapses may be rich in synaptic vesicles on the nerve fibers (presynaptic) side, whereas the cells (postsynaptic) side may contain a subsynaptic cistern; a flat compartment of the smooth endoplasmic reticulum. This structure is regarded as a prerequisite of a typical efferent synapse, as occurring in cochlear and vestibular hair cells. In fish taste buds, efferent synapses are rare and were found only in a few species that belong to different taxa. The significance of efferent synapses in fish taste buds is not well understood, because efferent connections between the gustatory nuclei of the medulla with taste buds are not yet proved. PMID- 16217621 TI - Protocols for two- and three-color fluorescent RNA in situ hybridization of the main and accessory olfactory epithelia in mouse. AB - The main and accessory olfactory epithelia of the mouse are composed of many cell populations. Each sensory neuron is thought to express one allele of one of the approximately 1000 odorant or approximately 300 vomeronasal receptor genes. Sensory neurons die and are replaced by new neurons that differentiate from precursor cells throughout the lifetime of the individual. Neuronal replacement is asynchronous, resulting in the co-existence of cells at various stages of differentiation. Receptor gene diversity and ongoing neuronal differentiation produce complex mosaics of gene expression within these epithelia. Accurate description of gene expression patterns will facilitate the understanding of mechanisms of gene choice and differentiation. Here we report a detailed protocol for two- and three-color fluorescent RNA in situ hybridization (ISH) and its combination with immunohistochemistry, or detection of bromodeoxyuridine (BrdU) incorporated DNA after labeling. The protocol is applied to cryosections of the main and accessory olfactory epithelia in mouse. PMID- 16217622 TI - Homer 1b/c expression correlates with zebrafish olfactory system development. AB - The zebrafish, (Danio rerio) is an important model organism for the analysis of molecular mechanisms that govern neuronal circuit development. The neuronal circuitry that mediates olfaction is crucial for the development and survival of all teleost fishes. In concert with other sensory systems, olfaction is functional at early stages in zebrafish development and mediates important behavioral and survival strategies in the developing larva. Odorant cues are transduced by an array of signaling molecules from receptors in olfactory sensory neurons. The scaffolding protein family known as Homer is well placed to orchestrate this signaling cascade by interacting with and coupling membrane bound receptors to cytosolic signaling partners. To date, Homer has not been demonstrated in the zebrafish. Here we report that the Homer 1b/c isoform was prominent in the olfactory system from the earliest stages of differentiation. We describe the spatial and temporal distribution of Homer in the zebrafish olfactory system. At 24 hours post fertilization (hpf), Homer expression delineated the boundary of the presumptive olfactory placode. Subsequent expression steadily increased throughout the developing olfactory placode, with a prominent localization to the dendritic knobs of the olfactory sensory neurons. Homer expression in the developing olfactory bulb was punctate and prominent in the glomeruli, displaying an apparent synaptic localization. This work supports the hypothesis that Homer is an important molecule in neuronal circuit development, necessary for crucial behaviors required for development and survival. PMID- 16217623 TI - Transient expression of tyrosine hydroxylase promoter/reporter gene constructs in the olfactory epithelium of transgenic mice. AB - Maturation and survival of olfactory receptor neurons (ORNs) are hypothesized to depend on trophic support from the olfactory bulb during both development and regeneration of the olfactory epithelium (OE). The current study characterized transgene expression in two independently derived transgenic mouse lines in which 9 kb of tyrosine hydroxylase (TH) promoter was utilized to drive either enhanced green fluorescent protein (TH/eGFP) or LacZ (TH/beta-gal) reporters. Transgene expression, found primarily on dorsal aspects of the OE, the dorsal septum and endoturbinate II, resembled the Zone one distribution of olfactory receptor genes. Labeled cells were ovoid to fusiform with dendrites that projected to the epithelial surface but only rarely exhibited discernable cilia. Axons were short and did not extend beyond the basal lamina. As only a subpopulation of the cells contained olfactory marker protein, indicative of ORN maturation, the transgene expressing cells were likely immature neuronal precursors. Demonstration of transgene expression without either TH mRNA or protein was consistent with low basal level transcriptional activity of endogenous TH that may reflect differences between TH and reporter protein stability. Molecules identifying specific olfactory-derived cell populations, PDE2 and LHRH, also did not co localize with either reporter. A higher than predicted proportion of apoptotic neonatal transgene-expressing cells accounted for their apparent paucity in adult mice. These studies support the concept that transgene expressing cells exhibiting morphological and biochemical characteristics of presumptive ORNs are unable to mature and undergo apoptotic cell death possibly because they lack trophic support. PMID- 16217624 TI - Fine structural aspects of apoptosis in the olfactory epithelium. AB - Vertebrate olfactory receptor neurons are unique because they are continually replaced throughout life. They die by apoptosis under physiological conditions at all stages in their life cycle, and the dead olfactory neurons are replaced by the progeny of dividing basal cells. Thus, in the olfactory epithelium apoptosis is involved in tissue homeostasis and may be a direct or indirect trigger of neurogenesis. In this study, we focused on morphological changes occurring in the olfactory epithelium, i.e., degradation of DNA, condensation of nuclear chromatin, condensation of cytoplasm, blebbing of cytoplasmic fragments, and disposal of the dying and dead cells as the final phase of apoptosis. Moreover, we addressed other stages of apoptosis examining the nature of the stimulus that provokes the apoptotic response, the signal or metabolic state, and transduction of the signal that sends the message to the effector apparatus, and the effector or execution phase, which includes the activation of proteases. PMID- 16217625 TI - The role of relatives in discharge planning from psychiatric hospitals: the perspective of patients and their relatives. AB - This study prospectively assessed the preferences and satisfaction of 98 psychiatric inpatients and 40 of their relatives with family involvement in discharge planning. Preferences questionnaires were administered during hospitalization. Satisfaction questionnaires were completed 3 months later. Preferences noted by most participants included information concerning patient health status, ways to prevent further hospitalizations, services for relatives, and signs of patient decompensation. More relatives than patients felt that post discharge residence and activities were important areas to be involved in. Most participants were satisfied if relatives were involved in discharge planning. However, up to 89% of patients, and 84% of relatives, reported no communication between clinical staff and relatives regarding discharge. When this was the case, satisfaction rates dropped sharply, especially for relatives. The need for increased communication between clinicians and relatives regarding discharge planning remains a problem. PMID- 16217626 TI - Precipitants to psychiatric patient assaults on staff: review of empirical findings, 1990-2003, and risk management implications. AB - Psychiatric patient assaults on staff are a serious and continuing problem in health care settings. Over thirty years of empirical research have documented the characteristics of both patient assailants and staff victims. Notably absent from this literature have been similar empirical studies on the nature of patient precipitants to these assaults. This paper reviewed empirical studies of patient assault precipitants from 1990-2003. Six studies from three countries were reviewed. Common precipitants included staff restrictions on patient behaviors, denial of services, excessive sensory overload, and provocation by others. The clinical, methodological, and risk management implications are examined. PMID- 16217629 TI - Treatment of the stress casualty during operation Iraqi freedom one. AB - INTRODUCTION: Service members with mental health issues during Operation Iraqi Freedom receive evaluation and treatment at different echelons of care. Mental health teams consist of professionals and paraprofessionals assigned to Combat Stress Control units, Division Mental Health Sections, and Combat Support Hospitals. METHODS: Army doctrine for mental health care is delineated and examples describing how mental health personnel must be flexible in this challenging environment and are useful mental health tools for commanders in the operational environment are presented. DISCUSSION: Specific treatment issues related to the combat environment during Operation Iraqi Freedom are discussed in relation to each team and in light of doctrine. PMID- 16217630 TI - Psychiatric interventions with returning soldiers at Walter Reed. AB - War is a malefic force and results in many psychiatric and medical casualties. Psychiatry's involvement with soldiers experiencing psychological stress resulting from combat experience has been reported for many years (Zajtchuk, 1995). It has been demonstrated that a myriad of diagnosis to include depression, anxiety, somatoform, adjustment disorders and psychotic behaviors also emerge (Wain et al., 1996, 2005a). Nearly all survivors exposed to traumatic events briefly exhibit one or more stress related symptoms (Morgan et al., 2003). In many instances these symptoms dissipate within a reasonable amount of time. However, symptoms persisting for a prolonged period following a traumatic event increase the probability of developing stress-related psychiatric disorders. PMID- 16217627 TI - A rating scale for disruptive behavior disorders, based on the DSM-IV item pool. AB - DSM IV includes three clusters of items that are used to establish diagnoses for the Disruptive Behavior Disorders: Attention Deficit, Conduct, and Oppositional Defiant. In this report, we examine the feasibility of using the items in each cluster to form a rating scale. We studied eighty-four consecutive school-aged referrals to an inner-city child and adolescent Psychiatry clinic. Case diagnosis was established with a clinician's KID-SCID assessment. Parents and teachers rated the 41 DSM items on four-point scales, and completed the Conners' Rating Scales, in English or Spanish. In this paper we report psychometrics of the new scale, the Rating Scale for Disruptive Behavior Disorders (RS-DBD), along with the agreement among parents and teachers, and concurrence between the new scales and the relevant Conners' scales. While, the parent and teacher ratings may provide a useful index for severity of behavioral disturbance in the home and school environments, it will not establish a diagnosis. There was a great deal of comorbidity among diagnostic groups. PMID- 16217631 TI - Re-entry and reintegration: returning home after combat. AB - Soldier life exists on a continuum of readiness for deployment. Re-entry and reintegration-the return home and reunion with family and community-key the success of the deployment cycle. In current and projected future operations, the Army and society will both bear the burden of this re-entry and re-integration. Programs and procedures in place work towards improving communication, mitigating distress and resolving crises during reentry and reintegration. Key elements include: inclusion of families and communities early into the planning for reentry and reintegration; normalization (non-medicalization of distress); easy access to behavioral health professionals; and education of families on resources and benefits. Through broad collaboration, maximal benefit to the Soldier, family members and society be realized. PMID- 16217632 TI - Military families and children during operation Iraqi freedom. AB - The general public has become increasingly interested in the health and well being of the children and families of military service members as the war in Iraq continues. Observers recognize the potential stresses or traumas that this population might undergo as a result of the military deployment or the possible injury or death of military family members. While such concern is welcomed, it is sometimes misplaced. Not infrequently, conclusions that are drawn are fraught with misunderstanding and bias based upon lack of understanding of the military community or a preconceived notion of the vulnerabilities of the population. This problem is compounded by the paucity of scientific study. In this article the authors review the strengths of military families as well as the unique challenges that they face. The authors also highlight parental deployment, parental injury and parental death as unique stresses to military children and families. Available and pertinent scientific information is reviewed. Clinical observations of children and families during the ongoing war in Iraq are presented. PMID- 16217634 TI - [The health of unemployed women and men. First results of the Telephone Health Survey 2003]. AB - Unemployment has material and psychosocial consequences that can also have an adverse effect on health. According to the Telephone Health Survey 2003, unemployed women and men suffer more frequently from many diseases and complaints. Their assessment of their own state of health is also more negative, and they say more often that their quality of life is restricted by their health. In the case of men, health problems are especially common among the long-term unemployed (>1 year), whereas women can suffer equally (or even more) serious effects on their health as a result of short-term unemployment. On the other hand, the analysis clearly indicates that poor health heightens a person's risk of becoming unemployed. However, no conclusive statement can be made on the direction of causality between unemployment and health, since this requires more longitudinal data. PMID- 16217635 TI - [Diabetes mellitus in Germany. Review of the situation according to the 2003 Telephone Health Survey]. AB - Diabetes mellitus is one of the most prevalent endocrine disorders in Germany. Secondary diseases and complications have serious effects on the lives of diabetics. They also represent an important challenge for economic and health policy processes. According to the 2003 Telephone Health Survey, the prevalence of diabetes mellitus among the population aged 18 years and older is 6.1% for women and 5.4% for men. The percentage of diabetics among older population groups is considerably higher (65+: 16%). The need for effective health care for diabetes patients is reflected in frequent medical consultations. At the same time, there are deficiencies in the treatment of diabetes, for example in HbA1c measurement and the examination of diabetics' ocular fundus and feet. Although special training courses for diabetics are well publicized, only one in two diabetics makes use of them. Future surveys will show whether disease management programmes can help bridge the existing health care gaps. PMID- 16217633 TI - A framework to analyze multiple time series data: a case study with Streptomyces coelicolor. AB - Transcriptional regulation in differentiating microorganisms is highly dynamic involving multiple and interwinding circuits consisted of many regulatory genes. Elucidation of these networks may provide the key to harness the full capacity of many organisms that produce natural products. A powerful tool evolved in the past decade is global transcriptional study of mutants in which one or more key regulatory genes of interest have been deleted. To study regulatory mutants of Streptomyces coelicolor, we developed a framework of systematic analysis of gene expression dynamics. Instead of pair-wise comparison of samples in different combinations, genomic DNA was used as a common reference for all samples in microarray assays, thus, enabling direct comparison of gene transcription dynamics across different isogenic mutants. As growth and various differentiation events may unfold at different rates in different mutants, the global transcription profiles of each mutant were first aligned computationally to those of the wild type, with respect to the corresponding growth and differentiation stages, prior to identification of kinetically differentially expressed genes. The genome scale transcriptome data from wild type and a DeltaabsA1 mutant of Streptomyces coelicolor were analyzed within this framework, and the regulatory elements affected by the gene knockout were identified. This methodology should find general applications in the analysis of other mutants in our repertoire and in other biological systems. PMID- 16217637 TI - [Clinical, radiological and arthroscopical aspects in nail patella syndrome. Literature review based on an affected family]. AB - Nail patella syndrome (NPS) is an autosomal dominant hereditary disorder affecting the nails, skeletal system, kidneys, and eyes. Skeletal features include absent or hypoplastic patellae, patella dislocations, elbow abnormalities, talipes and iliac horns on plain films. The existing literature focuses on clinical and radiographic findings in patients with NPS. We also report the case of a 40-year-old male patient and his family affected by NPS and includes clinical, radiographic as well as arthroscopic findings. Arthroscopic findings in this case are characterized by multiple synovial plicae in the knee joint leading to cartilage defects on the corresponding cartilage surfaces. A review of the recent literature suggests that the occurrence of synovial plicae might be related to NPS and might account for at least part of the characteristic symptoms of these patients. Therefore, in cases of unspecific knee pain associated with NPS arthroscopy of the knee joints and plicae resection seems to be recommendable. PMID- 16217638 TI - [Comparison of 2D and 3D sequences for MRCP. Clinical value of the different techniques]. AB - Magnetic resonance cholangio-pancreaticograpy (MRCP) is a non-invasive imaging modality of the pancreatico-biliary system which plays an increasingly important role in the clinical and diagnostic workup of patients with biliary or pancreatic diseases. The present review is designed to give an overview of the currently available and appropriate sequences, their technical background, as well as new developments and their relevance to the various clinical issues and challenges. The impact of the latest technical innovations, such as integrated parallel imaging techniques and navigator-based respiratory triggering, on the diagnostic capacities of MRCP is discussed. In this context, the individual value of RARE, T2w single shot turbo/fast spin echo (SSFSE) and the recently introduced 3D T2w turbo/fast spin echo sequences (T2w 3D-T/FSE) is reviewed. RARE imaging may be preferred in severely ill patients with limitations in cooperation, SSFSE is particularly effective in differentiating benign and malignant stenosis, and 3D FSE offers additional advantages in the detection of small biliary concrements. PMID- 16217640 TI - VQ scintigram in the CT pulmonary angiogram era. PMID- 16217641 TI - Single-site mutations on the catalase-peroxidase from Sinorhizobium meliloti: role of the distal Gly and the three amino acids of the putative intrinsic cofactor. AB - KatB is the only catalase-peroxidase identified so far in Sinorhizobium meliloti. It plays a housekeeping role, as it is expressed throughout all the growth phases of the free-living bacterium and also during symbiosis. This paper describes the functional and structural characterization of the KatB mutants Gly303Ser, Trp95Ala, Trp95Phe, Tyr217Leu, Tyr217Phe and Met243Val carried out by optical and electron spin resonance spectroscopy. The aim of this work was to investigate the involvement of these residues in the catalatic and/or peroxidatic reaction and falls in the frame of the open dispute around the factors that influence the balance between catalatic and peroxidatic activity in heme enzymes. The Gly303 residue is not conserved in any other protein of this family, whereas the Trp95, Tyr217 and Met243 residues are thought to form an intrinsic cofactor that is likely to play a role in intramolecular electron transfer. Spectroscopic investigations show that the Gly303Ser mutant is almost similar to the wild-type KatB and should not be involved in substrate binding. Mutations on Trp95, Tyr217 and Met243 clear out the catalatic activity completely, whereas the peroxidatic activity is maintained or even increased with respect to that of the wild-type enzyme. The k (cat) values obtained for these mutants suggest that Trp95 and Tyr217 form a huge delocalized system that provides a pathway for electron transfer to the heme. Conversely, Met243 is likely to be placed close to the binding site of the organic molecules and plays a crucial role in substrate docking. PMID- 16217642 TI - The role of histidine 200 in MndD, the Mn(II)-dependent 3,4 dihydroxyphenylacetate 2,3-dioxygenase from Arthrobacter globiformis CM-2, a site directed mutagenesis study. AB - The manganese-dependent 3,4-dihydroxyphenylacetate 2,3-dioxygenase (MndD) from Arthrobacter globiformis CM-2 is an extradiol-cleaving catechol dioxygenase that catalyzes aromatic ring cleavage of 3,4-dihydroxyphenylacetate (DHPA). Based on the recent crystal structure of the MndD-DHPA complex, a series of site-directed mutations were made at a conserved second-sphere residue, histidine 200, to gain insight into and clarify the role this residue plays in the Mn(II)-dependent catalytic mechanism. In this study, we report the activities and spectroscopic data of these H200 variants and their DHPA and 4-nitrocatechol (4-NC) complexes. The data collected from wild-type and mutant MndDs are consistent with a role for H200 interacting with a manganese-bound dioxygen moiety and are inconsistent with other previously proposed roles involving proton transfer. Spectroscopic observations, including unique low-field EPR signals found when DHPA and 4-NC are bound to the Mn(II) center of MndD, are discussed and their relationship to dioxygen activation catalyzed in MndD is explored. PMID- 16217643 TI - Centrally mediated antihyperalgesic and antiallodynic effects of zonisamide following partial nerve injury in the mouse. AB - Some antiepileptic drugs are used clinically to relieve neuropathic pain. We have evaluated the effects and investigated the possible mechanisms of action of zonisamide, an antiepileptic drug, on thermal hyperalgesia and tactile allodynia in a murine chronic pain model that was prepared by partial ligation of the sciatic nerve. Subcutaneously administered zonisamide (10 and 30 mg/kg) produced antihyperalgesic and antiallodynic effects in a dose-dependent manner; these effects were manifested by elevation of the withdrawal threshold in response to a thermal (plantar test) or mechanical (von Frey) stimulus, respectively. Similar analgesic effects were obtained in both the plantar and von Frey tests when zonisamide was injected either intracerebroventricularly (i.c.v., 10 and 30 microg) or intrathecally (i.t., 10 and 30 microg). It is thought that this elevation of the thermal and mechanical withdrawal thresholds after local injection of zonisamide is not generated secondarily via impaired motor activity, since zonisamide (30 microg, i.c.v. or i.t.) did not affect locomotor activity, as assessed in sciatic-nerve-ligated mice. Moreover, the nitric oxide synthase inhibitor L-NAME, when injected either i.c.v. or i.t., potentiated the analgesic effects of zonisamide. In contrast, neither i.c.v. nor i.t. zonisamide produced antinociceptive effects against acute thermal and mechanical nociception in non ligated mice. Together, following peripheral nerve injury, it appears that zonisamide produces centrally mediated antihyperalgesic and antiallodynic effects partly via the blockade of nitric oxide synthesis. PMID- 16217644 TI - Space radiation does not induce a significant increase of intrachromosomal exchanges in astronauts' lymphocytes. AB - Chromosome aberration analysis in astronauts has been used to provide direct, biologically motivated estimates of equivalent doses and risk associated to cosmic radiation exposure during space flight. However, the past studies concentrated on measurements of dicentrics and translocations, while chromosome intrachanges (inversions) have never been measured in astronauts' samples. Recent data reported in the literature suggest that densely ionizing radiation can induce a large fraction of intrachanges, thus leading to the suspicion that interchanges grossly underestimate the cosmic radiation-induced cytogenetic damage in astronauts. We have analyzed peripheral blood lymphocytes from 11 astronauts involved in short- or long-term space flights in low-Earth orbit using high-resolution multicolor banding to assess the frequency of intrachromosomal exchanges in both pre- and post-flight samples. We did not detect any inversions in chromosome 5 from a total of 2800 cells in astronauts' blood. In addition, no complex type exchanges were found in a total of 3590 astronauts' lymphocytes analyzed by multifluor fluorescence in situ hybridisation. We conclude that, within the statistical power of this study, the analysis of interchanges for biological dosimetry in astronauts does not significantly underestimate the space radiation-induced cytogenetic damage, and complex-type exchanges or intrachanges have limited practical use for biodosimetry at very low doses. PMID- 16217646 TI - A model of the coupling between grip aperture and hand transport during human prehension. AB - It has been repeatedly demonstrated that the opening between the index finger and thumb (grasp component) during an object-directed reach-to-grasp movement achieves maximum aperture approximately two-thirds of the way through the duration of the reaching movement (transport component). Here we offer a quantitative model of the temporal coupling between grip aperture and wrist velocity which shows experimentally that the correlation between grip aperture and object size is a sigmoidal function of movement duration. When wrist velocity reaches its peak value, the correlation between the grip aperture and the size of the goal object has reached half of the correlation that is achieved by the end of the movement. PMID- 16217645 TI - Seeing speech affects acoustic information processing in the human brainstem. AB - Afferent auditory processing in the human brainstem is often assumed to be determined by acoustic stimulus features alone and immune to stimulation by other senses or cognitive factors. In contrast, we show that lipreading during speech perception influences early acoustic processing. Event-related brainstem potentials were recorded from ten healthy adults to concordant (acoustic-visual match), conflicting (acoustic-visual mismatch) and unimodal stimuli. Audiovisual (AV) interactions occurred as early as approximately 11 ms post-acoustic stimulation and persisted for the first 30 ms of the response. Furthermore, the magnitude of interaction depended on AV pairings. These findings indicate considerable plasticity in early auditory processing. PMID- 16217647 TI - Lipid membrane-induced optimization for ligand-receptor docking: recent tools and insights for the "membrane catalysis" model. AB - Cells in living organisms are regulated by chemical and physical stimuli from their environment. Often, ligands interact with membrane receptors to trigger responses and Sargent and Schwyzer conceived a model to describe this process, "membrane catalysis". There is a notion that the physical organization of membranes can control the response of cells by speeding up reactions. We revisit the "membrane catalysis" model in the light of recent technical, methodological and theoretical advances and how they can be exploited to highlight the details of membrane mediated ligand-receptor interactions. We examine the possible effects that ligand concentration causes in the membrane catalysis and focus our attention in techniques used to determine the partition constant. The hypothetical diffusional advantage associated with membrane catalysis is discussed and the applicability of existing models is assessed. The role of in depth location and orientation of ligands is explored emphasizing the contribution of new analysis methods and spectroscopic techniques. Results suggest that membranes can optimize the interaction between ligands and receptors through several different effects but the relative contribution of each must be carefully investigated. We certainly hope that the conjugation of the methodological and technical advances here reported will revive the interest in the membrane catalysis model. PMID- 16217648 TI - Paramagnetic liposomes containing amphiphilic bisamide derivatives of Gd-DTPA with aromatic side chain groups as possible contrast agents for magnetic resonance imaging. AB - Three amphiphilic DTPA bisamide derivatives containing long-chain phenylalanine esters (with 14, 16 and 18 carbon atoms in the alkyl chain) were synthesized and their corresponding gadolinium(III) complexes were prepared. The attempts to form paramagnetic micelles carrying the gadolinium(III) complexes yielded unstable or polydisperse micelles implying that the presence of the bulky aromatic side groups in the amphiphilic Gd-DTPA bisamide complexes results in an inefficient packing of the paramagnetic complex into micelles. All complexes were efficiently incorporated into liposomes consisting of 1,2-dipalmitoyl-sn-glycero-3 phosphocholine (DPPC), yielding stable and monodisperse paramagnetic liposomes. All liposomes had a comparable size, typically between 120 and 160 nm. As a result of the reduced mobility of the gadolinium(III) complexes, solutions of these supramolecular structures show a higher relaxivity than solutions of Gd DTPA. However, the relaxivity gain is lower compared to compounds consisting of purely aliphatic chains of the same length, most likely due to the less efficient packing or increased local mobility of the gadolinium(III) complex. In the case of the Gd-DTPA bisamide complex with 18 carbon atoms, the immobilization inside the liposomal structure is less effective, probably because the aliphatic chains of the complex are longer than the alkyl chains of the DPPC host, resulting in a relatively high local mobility. The paramagnetic liposomes containing the Gd-DTPA bisamide complexes with 14 carbon atoms showed the highest relaxivity because the optimal length match between the hydrophobic chains of the DPPC and the ligand allowed very efficient packing of the paramagnetic complex into the liposome. PMID- 16217649 TI - Protein self-association in the cell: a mechanism for fine tuning the level of macromolecular crowding? AB - A new role for protein self-association in the cell is discussed. An argument is advanced that when cellular protein is in its associated state the excluded volume component of the solution is minimized. Conversely, when cellular protein is in its dissociated state the excluded volume component of the solution is maximized. For proteins that make up a substantial fraction of the intracellular protein concentration, control of the self-association event thus presents itself as a means of regulating cellular processes that are influenced by different levels of volume exclusion. In this communication we examine how the control of protein association/dissociation might influence one such important process, namely the folding of a protein to a compact state. PMID- 16217650 TI - Macrophages during retina and optic nerve development in the mouse embryo: relationship to cell death and optic fibres. AB - We compared the spatial and temporal patterns of distribution of macrophages, with patterns of naturally occurring cell death and optic fibre growth during early retina and optic nerve development, in the mouse. We used embryos between day 10 of embryogenesis (E10; before the first optic fibres are generated in the retina) and E13 (when the first optic fibres have crossed the chiasmatic anlage). The macrophages and optic axons were identified by immunocytochemistry, and the apoptotic cells were detected by the TUNEL technique, which specifically labels fragmented DNA. Cell death was observed in the retina and the optic stalk long before the first optic axons appeared in either region. Subsequently, specialized F4/80-positive phagocytes were detected in chronological and topographical coincidence with cell death, which disappeared progressively. As development proceeded, the pioneer ganglion cell axons reached the regions where the macrophages were located. As the number of optic fibres increased, the macrophages disappeared. Therefore, cell death, accompanied by macrophages, preceded the growth of fibres in the retina and the optic nerve. Moreover, these macrophages synthesized NGF and the optic axons were p75 neurotrophin receptor (p75(NTR))- and TrkA-positive. These findings suggest that macrophages may be involved in optic axon guidance and fasciculation. PMID- 16217652 TI - Continuous production of L(+)-tartaric acid from cis-epoxysuccinate using a membrane recycle reactor. AB - The one-step bioconversion of cis-epoxysuccinate (CES) to L(+)-tartaric acid by dried Rhodococcus rhodochrous cells containing CES hydrolase activity was studied by using a continuous bioconversion process. The influence of the pH and the temperature was assessed. A mathematical model was used to quantify the CES hydrolase activity and stability. The optimal pH, which resulted in a maximal CES hydrolase activity and stability, was pH 8.0. A large increase in stability (half life time) could be obtained when the temperature was decreased from 37 to 14 degrees C during the continuous bioconversion. A total bioconversion was maintained for more than 100 days. This resulted in a large value for the specific productivity since the effect of the large increase in stability was much more important than the decrease of activity at the lower temperature. This continuous bioconversion process was further optimised by calculating the productivity for several continuously stirred tank reactors in series. The specific productivity could be nearly doubled when the number of reactors in the series was increased from 1 to 4. PMID- 16217651 TI - The nucleolar structure and the activity of NopA100, a nucleolin-like protein, during the cell cycle in proliferating plant cells. AB - For the purpose of gaining knowledge of the relationships between cell proliferation and ribosome biogenesis, as two fundamental mutually interconnected cellular processes, studies were performed on cell populations synchronized in their cell-cycle progression by treatment with hydroxyurea, followed by sampling at different times after its removal. A structural rearrangement of the nucleolus was observed throughout the interphase, along with changes in the relative amounts of different nucleolar subcomponents. A structural model of nucleolar organization was associated with each interphase period. Throughout interphase, the nucleolin-like protein, NopA100, was immunodetected in the dense fibrillar component of the nucleolus, preferentially near fibrillar centers and its levels were shown to increase from G1 to G2. A western blotting analysis of soluble nuclear protein extracts with anti-NopA100 antibody resulted in the intense labeling of a 100-kDa band, but also of a series of proteins related to it, suggesting that NopA100 undergoes a physiological process of proteolytic maturation, similar to that described for mammalian nucleolin, but not reported in other biological model systems. Physiological proteolysis of NopA100, related to cell-cycle progression, was confirmed after the nuclei extracted from synchronized cells were treated with the protease inhibitor, leupeptin, which resulted in an increase of the 100-kDa band at the expenses of the decrease of some other bands, according to the cell-cycle stages. We therefore conclude that there is a relationship between the increase in nucleolar activity, cell-cycle progression, nucleolar structure, the activity of NopA100, and the proteolysis of this nucleolin-like protein. PMID- 16217653 TI - Expression, purification, and immobilization of His-tagged D-amino acid oxidase of Trigonopsis variabilis in Pichia pastoris. AB - High-level expression of D: -amino acid oxidase (DAO) has been reported in Pichia pastoris by integrating the DAO gene under the control of the alcohol oxidase promoter (PAOX1). However, the time taken to reach peak product concentration is usually long (approximately 43 h), and cultivation requires tight regulation of methanol feeding. In this paper, we describe the expression of His-tagged DAO (HDAO) in P. pastoris using the glyceraldehydes-3-phosphate dehydrogenase promoter (PGAP). The maximal level of HDAO expression using the PGAP integrant is attained in 13 h and is equal to that obtained using the PAOX1 integrant in 43 h. We also explored the possibility of secreting HDAO in P. pastoris. In-frame fusion of Saccharomyces cerevisiae alpha-factor secretion signal under a PGAP or PAOX1 resulted in low-level secretion of active HDAO, which was not of practical use. The intracellularly expressed HDAO under PGAP was purified by agar-based affinity support and then immobilized on Amberzyme oxirane resin. The immobilized HDAO, with specific activity of 75 U g-1 (wet weight), could be recycled more than 14 times without significant loss of activity. The data suggest that intracellular production of HDAO under PGAP, followed by affinity purification and immobilization on oxirane resin, may serve as an effective process for the manufacture of immobilized DAO for industrial application. PMID- 16217654 TI - Active-site engineering of biphenyl dioxygenase: effect of substituted amino acids on substrate specificity and regiospecificity. AB - Biphenyl dioxygenase (Bph Dox) catalyzes the initial dioxygenation step in the metabolism of biphenyl. The large subunit (BphA1) of Bph Dox plays a crucial role in the determination of the substrate specificity of biphenyl-related compounds including polychlorinated biphenyls (PCBs). Previously, the substitution of Asn at Thr-376 near the active-site iron in the BphA1 of Pseudomonas pseudoalcaligenes KF707 expanded the oxidation range and altered the regiospecificity of Bph Dox for PCBs. In this study, we replaced Thr-376 with Gly, Ser, Gln, Tyr, Val, Phe, Asp, and Lys and expressed these enzymes in Escherichia coli. Bph Dox mutants of Thr376Asn, Thr376Val, Thr376Phe, and Thr376Lys showed novel degradation activity for dibenzofuran, which is a poor substrate for KF707 Bph Dox. All active Bph Dox mutants showed altered regiospecificity with 2,2'-dichlorobiphenyl and 2,5,4'-trichlorobiphenyl. The Thr376Gly, Thr376Val, Thr376Phe, and Thr376Asp Bph Dox mutants introduced molecular oxygen at the 2,3 position of 2,2'-dichlorobiphenyl, forming 2-chloro 2',3'-dihydroxybiphenyl with concomitant dechlorination. The Bph Dox mutants of Thr376Gly, Thr376Ser, Thr376Asp, and Thr376Lys attacked 2,5,4'-trichlorobiphenyl via both 2',3'- and 3,4-dioxygenation activities. In particular, the Thr376Phe Bph Dox mutant exhibited enhanced and expanded degradation activities toward all of the compounds tested. Further site-directed mutation was induced to change the oxidizing character of KF707 Bph Dox to that of the Bph Dox of Burkholderia xenovorans LB400 by the substitution of two amino acids, Ile335Phe and Thr376Asn, near the active-site. PMID- 16217655 TI - Molecular detection of the clostridia in an anaerobic biohydrogen fermentation system by hydrogenase mRNA-targeted reverse transcription-PCR. AB - Molecular biological approaches were developed to monitor the potential biohydrogen-producing clostridia in an anaerobic semisolid fermentation system that used brewery yeast waste as the fermentation substrate. The denaturing gradient gel electrophoresis with 16S rDNA gene-targeted polymerase chain reaction (PCR) analysis was employed to confirm the existence of clostridia in the system. Remarkably, reproducible nucleotide sequences of clostridia were obtained from different hydrogen production stages by using hydrogenase gene targeted reverse transcription (RT)-PCR. These RNA-based information suggested that the predominant hydrogen-producing strains possess either a specific Clostridium pasteurianum-like or a specific Clostridium saccharobutylicum-like hydrogenase sequence. Comparison of the hydrogenase gene-targeted sequence profiles between PCR and RT-PCR revealed that the specific C. pasteurianum-like hydrogenase harboring bacterial strains were dominant in both mRNA and bacterial population level. On the other hand, the specific C. saccharobutylicum-like hydrogenase harboring strains expressed high level of hydrogenase mRNA but may not be dominant in population. Furthermore, quantitative real-time RT-PCR analysis showed the expression pattern of the clostridial hydrogenase mRNA and may serve as an activity index for the system. PMID- 16217656 TI - Multi-objective optimization in Aspergillus niger fermentation for selective product enhancement. AB - A multi-objective optimization formulation that reflects the multi-substrate optimization in a multi-product fermentation is proposed in this work. This formulation includes the application of epsilon-constraint to generate the trade off solution for the enhancement of one selective product in a multi-product fermentation, with simultaneous minimization of the other product within a threshold limit. The formulation has been applied to the fed-batch fermentation of Aspergillus niger that produces a number of enzymes during the course of fermentation, and of these, catalase and protease enzyme expression have been chosen as the enzymes of interest. Also, this proposed formulation has been applied in the environment of three control variables, i.e. the feed rates of sucrose, nitrogen source and oxygen and a set of trade-off solutions have been generated to develop the pareto-optimal curve. We have developed and experimentally evaluated the optimal control profiles for multiple substrate feed additions in the fed-batch fermentation of A. niger to maximize catalase expression along with protease expression within a threshold limit and vice versa. An increase of about 70% final catalase and 31% final protease compared to conventional fed-batch cultivation were obtained. Novel methods of oxygen supply through liquid-phase H2O2 addition have been used with a view to overcome limitations of aeration due to high gas-liquid transport resistance. The multi objective optimization problem involved linearly appearing control variables and the decision space is constrained by state and end point constraints. The proposed multi-objective optimization is solved by differential evolution algorithm, a relatively superior population-based stochastic optimization strategy. PMID- 16217657 TI - The role of endocytosis in renal dopamine D1 receptor signaling. AB - Desensitization of G-protein-coupled receptors (GPCR) includes receptor endocytosis. This phenomenon is suggested, at least for some receptors, to be associated with receptor resensitization. Here, we examined the role of receptor endocytosis for two different GPCR, the dopamine-1 (D1) receptor and the beta1 adrenoceptor (beta(1)-AR) in renal tissue. The functional role of receptor endocytosis was examined on Na+, K+ -ATPase activity in microdissected proximal tubules from rat kidney. The spatial regulation of endogenous D1 receptors and beta(1)-AR was examined by confocal microscopy techniques in LLCPK cells. Phenylarsine oxide (PAO) an endocytosis inhibitor, attenuated isoproterenol induced decrease in Na+, K+ -ATPase activity but had no such effect on dopamine induced decrease in Na+, K+ -ATPase activity. We have previously shown that isoproterenol sensitizes the renal dopamine system, by recruiting silent D1 receptors from the interior of the cell towards the plasma membrane. This effect was attenuated by PAO as well as by cytochalasin D while these substances had no effect on dopamine-induced D1 receptor recruitment. The beta(1)-AR was localized to the plasma membrane in control cells. Isoproterenol induced a rapid internalization of the beta(1)-AR; which was prevented by PAO. The results suggest that endocytosis of beta(1)-AR in renal proximal tubular cells is an important step in signal generation, while endocytosis of proximal tubular D1 receptor is not. PMID- 16217658 TI - Ischemia-induced increase of stiffness of alphaB-crystallin/HSPB2-deficient myocardium. AB - The two small heat shock proteins (sHSPs), alphaB-crystallin and HSPB2, have been shown to translocate within a few minutes of cardiac ischemia from the cytosol to myofibrils; and it has been suggested that their chaperone-like properties might protect myofibrillar proteins from unfolding or aggregation during stress conditions. Further evidence of an important role for HSPs in muscle function is provided by the fact that mutations of the alphaB-crystallin gene cause myopathy and cardiomyopathy. In the present study, we subjected isolated papillary muscles of alphaB-crystallin/HSPB2-deficient mice to simulated ischemia and reperfusion. During ischemia in alphaB-crystallin/HSPB2-deficient muscles, the development of contracture started earlier and reached a higher value compared to the wildtype mice. The recovery of contracture of alphaB-crystallin/HSPB2-deficient muscles was also attenuated during the simulated reperfusion period. However, twitch force was not significantly altered at any time of the experiment. This suggests that during ischemic insults, alphaB-crystallin/HSPB2 may not be important for the contraction process itself, but rather serve to maintain muscular elasticity. PMID- 16217660 TI - Plasma thymulin and nerve growth factor levels in patients with primary open angle glaucoma and elevated intraocular pressure. AB - BACKGROUND: The objective was to measure the plasma concentrations of thymulin and nerve growth factor (NGF) in a group of patients with primary open angle glaucoma (POAG) and compare them with age- and sex-matched normal controls. METHODS: Twenty-eight patients newly diagnosed with POAG who were not undergoing treatment were compared with the same number of age- and sex-matched healthy controls. Blood samples were drawn into heparinized tubes and plasma samples were collected for the determination of the concentrations of thymulin and NGF, using specific enzyme-linked immunosorbent assay (ELISA). The Student's t test was used to perform the necessary statistical analysis of the results. RESULTS: Seventeen women and 11 men were enrolled in each of the two groups (study and control), with a mean age of 63.7 (SD 10.3) years in the former and 63.3 (SD 9.6) years in the latter. There was a highly significant (p<0.001) elevation in the thymulin levels in POAG patients compared with the control group. However, no significant difference was observed when comparing the plasma NGF levels. CONCLUSION: This is the first report to measure plasma thymulin levels in glaucoma patients. The significant results point the possible role of this immunomodulator in the pathogenesis of primary open angle glaucoma. The potential role of NGF seems to be less likely. These findings warrant further investigation. PMID- 16217659 TI - Incidence, causes and prognosis of hypotension related to meprobamate poisoning. AB - OBJECTIVE: Meprobamate self-poisoning has been reported as potentially inducing hypotension. We examined the incidence and causes of hypotension induced by this poisoning and its prognosis. DESIGN AND SETTING: Retrospective observational study conducted in a medical ICU between June 1997 and October 2003. Seventy-four patients admitted for meprobamate poisoning and needing mechanical ventilation were included. Demographic, clinical, and laboratory data were compared between patients with and without hypotension. All echocardiograms recorded in patients with hypotension were reviewed, and left ventricular (LV) and right ventricular (RV) functions were assessed. RESULTS: Twenty-nine (40%) patients exhibited hypotension without any significant difference in age, gender, cardiac history, or meprobamate concentration in blood when compared to patients without hypotension. Base excess was significantly lower in patients with hypotension. Echocardiography demonstrated a hypokinetic state, associating decreased LV ejection fraction (45+/-15%) and cardiac index (2+/-0.7 l min(-1) m(-2)), and increased inferior vena cava diameter. Most patients with hypotension received inotropic drugs by infusion, and were ventilated for significantly longer. CONCLUSIONS: Meprobamate self-poisoning induces hypotension, notably related to cardiac failure, in about 40% of cases. This has important therapeutic consequences, as frequent inotropic drug infusion. The mechanisms of cardiac toxicity remain largely unknown, and no predictive factor could be isolated. PMID- 16217661 TI - Alteration of tear cytokine balance by eye closure: analysis by multicytokine assay. AB - PURPOSE: To study the effects of eye closure on the balance of tear cytokines. METHODS: A total of 17 healthy volunteers with normal eyes were recruited. Tears (10 microl) were collected by capillary outflow at 09:00. The right eyes were closed with cotton bandages for 3 h, then tears were collected from each eye separately. The concentrations of inflammatory cytokines, interleukin (IL)-1beta, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor (TNF)-alpha were measured by cytometric bead array. RESULTS: Although the concentration of each cytokine varied between eyes, the increasing ratio for 3 h showed a similar pattern in each cytokine. IL-1beta, IL-6, IL-10, IL-12p70, and TNF-alpha increased and IL-8 decreased in normal eyes. After eye closure, the increase of IL-10 significantly diminished (P=0.037); however, the other cytokines were not affected. CONCLUSIONS: Eye closure decreased the concentration of IL-10 in tears, which might cause a potentially pro-inflammatory status of the ocular surface. PMID- 16217663 TI - Biomechanical characterization of the three-dimensional kinematic behaviour of the Dynesys dynamic stabilization system: an in vitro study. AB - The Dynesys, a flexible posterior stabilization system that provides an alternative to fusion, is designed to preserve intersegmental kinematics and alleviate loading at the facet joints. Recent biomechanical evidence suggests that the overall range of motion (ROM) with the Dynesys is less than the intact spine. The purpose of this investigation was to conduct a comprehensive characterization of the three-dimensional kinematic behaviour of the Dynesys and determine if the length of the Dynesys polymer spacer contributes to differences in the kinematic behaviour at the implanted level. Ten cadaveric lumbar spine segments (L2-L5) were tested by applying a pure moment of +/-7.5 Nm in flexion extension, lateral bending, and axial rotation, with and without a follower preload of 600 N. Test conditions included: (a) intact; (b) injury; (c) injury stabilized with Dynesys at L3-L4 (standard spacer); (d) long spacer (+2 mm); and (e) short spacer (-2 mm). Intervertebral rotations were measured using an optoelectronic camera system. The intersegmental range of motion (ROM), neutral zone (NZ), and three-dimensional helical axis of motion (HAM) were calculated. Statistical significance of changes in ROM, NZ, and HAM was determined using repeated measures analysis of variance (ANOVA) and Student-Newman-Keuls post-hoc analysis with P<0.05. Implantation of the standard length Dynesys significantly reduced ROM compared to the intact and injured specimens, with the least significant changes seen in axial rotation. Injury typically increased the NZ, but implantation of the Dynesys restored the NZ to a magnitude less that that of the intact spine. The Dynesys produced a significant posterior shift in the HAM in flexion-extension and axial rotation. The spacer length had a significant effect on ROM with the long spacer resulting in the largest ROM in all loading directions without a follower preload. The largest differences were in axial rotation. A 4 mm increase in spacer length led to an average intersegmental motion increase of 30% in axial rotation, 23% in extension, 14% in flexion, and 11% in lateral bending. There were no significant changes in NZ with different spacer lengths. Typically, the short spacer caused a greater shift and a greater change in orientation of the HAM than the long spacer. The long spacer resulted in a ROM and a motion pattern, as represented by the HAM, that was closer to that seen in an intact specimen. The results of this study suggest that the length of the Dynesys spacer altered the segmental position and therefore affected kinematic behaviour. PMID- 16217662 TI - Comparison of topical 0.7% dexamethasone-cyclodextrin with 0.1% dexamethasone sodium phosphate for postcataract inflammation. AB - BACKGROUND: To compare 0.7% dexamethasone-cyclodextrin aqueous eye drop solution applied once daily with 0.1% dexamethasone sodium phosphate eye drops applied three times a day for the control of postoperative inflammation after cataract surgery. METHODS: Twenty cataract patients who underwent phacoemulsification and intraocular lens implantation were randomly divided into two postoperative treatment groups. Postoperative medication in group I included 0.1% dexamethasone sodium phosphate eye drops three times daily and in group II 0.7% dexamethasone cyclodextrin eye drop solution once daily. Testing of visual acuity, biomicroscopic examination, applanation tonometry and laser flare cell meter (LFCM) examination were carried out before operation and days 1, 3, 7 and 21 after surgery. RESULTS: Preoperative and postoperative visual acuity, aqueous flare and cells in biomicroscopic examination, and the mean intraocular pressure did not show any statistically significant differences between the treatment groups. LFCM examination showed that the mean postoperative photon count values (P=0.032) and the median cell count values on the 1st (P=0.014), 3rd (P=0.031), 7th (P=0.034), and 21st (P=0.0097) postoperative days in group I were more elevated than in group II. CONCLUSIONS: 0.7% dexamethasone-cyclodextrin eye drops applied once daily is a more effective postoperative anti-inflammatory medication than 0.1% dexamethasone sodium phosphate applied three times a day. In both groups, 3 weeks after the operation the mean visual acuity was normal and intraocular pressure significantly lower than before operation. The use of 0.7% dexamethasone-cyclodextrin eye drops may be useful especially in elderly people who cannot apply themselves the eye drops onto the eye. PMID- 16217664 TI - A critical review of guidelines for low back pain treatment. AB - MAIN PROBLEM: Little is known about the methodological quality of guidelines for low back pain treatment. We evaluated the methods used by the developers according to established standards. METHODS: PubMed, guideline databases, and the World Wide Web were used to identify guidelines. Seventeen guidelines met the inclusion criteria: interventions for low back pain stated, recommendations based on or explicitly linked to evidence, and English version available. Guidelines were evaluated independently by two appraisers using a practical tool for this purpose, Users' Guides to the Medical Literature, and the Appraisal of Guidelines for Research and Evaluation (AGREE) instrument. RESULTS: Thirteen guidelines (76%) specified the most important therapies applied, but only nine (53%) included a complete description of the target population. Explicit processes to identify, select, and combine evidence were described in only six guidelines (35%). Few guidelines (3; 18%) explicitly considered all main outcomes when formulating therapeutic recommendations, and none contained a process to determine the relative value of different outcomes. Methodological criteria for grading the strength of the recommendations varied, and were often insufficiently specified. None of the guidelines assessed the impact of uncertainty associated with the evidence and values used. According to AGREE the quality score was highest for the scope and purpose, and clarity and presentation domains, and lowest for editorial independence and applicability. With regard to the recommendations, there was consensus for some of the interventions for acute pain (analgesics and NSAIDs, maintaining physical activity, and avoiding excessive bed rest), but explicit recommendations were lacking or ambiguous for 41% of the interventions. Most of the guidelines did not contemplate specific recommendations for chronic pain. CONCLUSIONS: A small number of the available guidelines for low back pain treatment achieved acceptable results for specific quality criteria. In general, the methods to develop the guidelines' therapeutic recommendations need to be more rigorous, more explicit and better explained. In addition, greater importance should be placed on the recommendations for chronic pain. PMID- 16217665 TI - Fusion for low-grade adult isthmic spondylolisthesis: a systematic review of the literature. AB - The objective of this study was to evaluate which fusion technique provides the best clinical and radiological outcome for adult low-grade lumbar isthmic spondylolisthesis, and to assess the overall clinical and radiological outcome of each fusion technique. A systematic review was performed. Medline, Embase, Current Contents, and Cochrane databases as well as reference lists of selected articles were searched. Randomised controlled trials (RCTs) were used to evaluate the best treatment; controlled studies and non-controlled studies were used to determine the outcomes after surgery. Two independent reviewers evaluated the studies with the methodological checklists of van Tulder and Jadad for the randomised studies and of Cowley for the non-randomised studies. The search resulted in 684 references and eventually 29 studies met the inclusion criteria, of which eight were RCTs, four were prospective, and 17 were retrospective case series. Ten of the case series did not clearly identify consecutive patient selection. All the eight RCTs evaluated the effect of different techniques of posterolateral fusion (PLF). Evidence was found that the PLF was superior to non operative treatment (exercise). Circumferential fusion was compared to PLF, but no difference could be found. PLF with or without instrumentation was evaluated in three studies, but no benefits from additional instrumentation were found. Other comparisons within PLF showed no effect of decompression, alternative instrumentation, or bone graft substitute. The 21 case series included 24 patient groups. PLF was used in 15 groups, good or excellent clinical outcome varied from 60 to 98% and fusion rate varied from 81 to 100%. Anterior interbody fusion was used in five groups, good or excellent clinical outcome varied from 85 to 94% and fusion rate varied from 47 to 90%. Posterior interbody fusion was used in two groups, good or excellent clinical outcome was 45% and fusion rate was 80 and 95%, respectively. Reduction, loss of reduction, and lordotic angles before and after the treatment was reported in only four studies. Average reduction achieved was 12.3%, average loss of reduction at follow-up was 5.9%. Preoperative lordotic angles were too heterogeneous to pool the results. Adjacent segment degeneration was not reported in any of the publications. A wide variety of complications were reported in 18 studies and included neurological complications, instrument failure, and infections. Fusion for low-grade isthmic spondylolisthesis has better outcomes than non-operative treatment. The current study could not identify the best surgical technique (PLF, PLIF, ALIF, instrumentation) to perform the fusion. However, instrumentation and/or decompression may play a beneficial role in the modern practice of reduction and fusion for low-grade isthmic spondylolisthesis, but there are no studies yet available to confirm this. The outcomes of fusion are generally good, but reports vary widely. PMID- 16217667 TI - Synchronous intracortical adamantinomas with keratin cyst formation. AB - Adamantinoma of the long bones is a rare primary bone tumor of uncertain embryogenesis. It tends to involve the tibia almost exclusively. We report on adamantinomas occurring in a 16-year-old male patient, with synchronous tibial and fibular lesions. Histologically, there were characteristic clusters of epithelial cells in a fibrous background, forming a keratin cyst. Immunohistochemically, these cells were strongly positive for cytokeratin. This keratin cyst formation is quite an unusual finding in classic adamantinoma. PMID- 16217666 TI - Effects of lumbar sympathectomy on pain behavioral changes caused by nucleus pulposus-induced spinal nerve damage in rats. AB - It has been suggested that lumbar sympathectomy can reduce pain behavior, including mechanical allodynia and thermal hyperalgesia, caused by ligation of the spinal nerve. One well-characterized model, which involves application of nucleus pulposus to the spinal nerve and displacement of the adjacent nerve, shows behavioral changes in rats. However, there have been no previous reports regarding sympathectomy performed in this model. Disk incision and adjacent spinal nerve displacement were performed with (n=6) or without (n=6) sympathectomy. Sham surgery was also performed with (n=6) or without (n=6) sympathectomy. The animals were tested for 3 days before surgery and on days 1, 3, 7, 14, and 21 after surgery. Non-noxious mechanical thresholds were tested by determining the hind paw withdrawal response to von Frey hair stimulation of the plantar surface of the footpad using a touch stimulator. Thermal nociceptive thresholds were tested using a sensitive thermal-testing device. While rats in the disk incision with displacement surgery group showed allodynia and hyperalgesia after surgery on the experimental side, sympathectomized animals did not. No allodynia was observed in the sham groups. Sympathectomy seemed to prevent the pain behavioral changes caused by the combination of disk incision and nerve displacement. PMID- 16217668 TI - The convoluted evolution of snail chirality. AB - The direction that a snail (Mollusca: Gastropoda) coils, whether dextral (right handed) or sinistral (left-handed), originates in early development but is most easily observed in the shell form of the adult. Here, we review recent progress in understanding snail chirality from genetic, developmental and ecological perspectives. In the few species that have been characterized, chirality is determined by a single genetic locus with delayed inheritance, which means that the genotype is expressed in the mother's offspring. Although research lags behind the studies of asymmetry in the mouse and nematode, attempts to isolate the loci involved in snail chirality have begun, with the final aim of understanding how the axis of left-right asymmetry is established. In nature, most snail taxa (>90%) are dextral, but sinistrality is known from mutant individuals, populations within dextral species, entirely sinistral species, genera and even families. Ordinarily, it is expected that strong frequency dependent selection should act against the establishment of new chiral types because the chiral minority have difficulty finding a suitable mating partner (their genitalia are on the 'wrong' side). Mixed populations should therefore not persist. Intriguingly, however, a very few land snail species, notably the subgenus Amphidromus sensu stricto, not only appear to mate randomly between different chiral types, but also have a stable, within-population chiral dimorphism, which suggests the involvement of a balancing factor. At the other end of the spectrum, in many species, different chiral types are unable to mate and so could be reproductively isolated from one another. However, while empirical data, models and simulations have indicated that chiral reversal must sometimes occur, it is rarely likely to lead to so-called 'single-gene' speciation. Nevertheless, chiral reversal could still be a contributing factor to speciation (or to divergence after speciation) when reproductive character displacement is involved. Understanding the establishment of chirality, the preponderance of dextral species and the rare instances of stable dimorphism is an important target for future research. Since the genetics of chirality have been studied in only a few pulmonate species, we also urge that more taxa, especially those from the sea, should be investigated. PMID- 16217669 TI - Antimalarial efficacy of methylene blue and menadione and their effect on glutathione metabolism of Plasmodium yoelii-infected albino mice. AB - Plasmodium yoelii infection caused significant decline in the hepatic and splenic glutathione content and the activities of the key enzymes, that is, glutamate cysteine ligase (EC 6.3.2.2) and glutathione reductase (EC 1.8.1.7) of their murine host, that is, Swiss albino mice. Methylene blue as well as menadione were found to restore these constituents when given to P. yoelii-infected mice at the dose levels of 2.5 and 100 mg/kg, respectively, compared to mefloquine which does the same at 5.0 mg/kg dose. Methylene blue, like mefloquine also caused a rapid decline in percent parasitaemia, whereas menadione caused a delay in maturation of the infection, but could not cure the mice. PMID- 16217670 TI - Posttraumatic anterior-inferior instability of the shoulder: arthroscopic findings and clinical correlations. AB - INTRODUCTION: The main purpose of our study was to evaluate intra-articular lesions in glenohumeral-instability with arthroscopy and correlate them with clinical findings as well as history of instability. MATERIAL AND METHODS: In this prospective multi-centre study, we evaluated arthroscopic findings in 303 patients with posttraumatic anterior-inferior instability of the shoulder. The study cohort was divided into 2 groups: patients with a history of one dislocation (Group 1, n=61, 20.1%) and patients with a history of more than one dislocation (Group 2, n=242, 79.9%). RESULTS: In Group 1, 37 patients had an IGHL lesion, 31 a MGHL-lesion and 41 a Hill-Sachs lesion. In Group 2, 182 patients had an IGHL-lesion, 172 a MGHL-lesion and 203 a Hill-Sachs lesion. The percentage of lesions in Group 2 (IGHL-75.2%, MGHL-71.1%, Hill-Sachs-83.9%) was significantly higher than in Group 1 (IGHL-60.7%, MGHL-50.8%, Hill-Sachs-67.2%, P=0.0233, P=0.0026, and P=0.0033, respectively). Within Group 2 we found significantly more Hill-Sachs-lesions with a history of an increasing number of recurrences (P=0.0436). We also found an increase of IGHL- and MGHL-lesions with an increasing number of recurrences, but this difference was not significant. The distribution of lesion types of the anterior labrum-ligament complex showed no significant difference between the two groups, apart from a higher incidence of ALPSA-lesions within Group 2 (34.7% versus 18.0% in Group 1). The results of this study show that recurrences after primary posttraumatic anterior-inferior shoulder dislocation cause increasing ligamental damage as well as increasing Hill-Sachs lesions within the gleno-humeral joint. CONCLUSION: Thus we conclude that early surgical stabilization after posttraumatic anterior-inferior shoulder dislocation is necessary to prevent increasing damage within the shoulder joint. PMID- 16217671 TI - Bifocal disruption of the knee extensor apparatus ("floating patella") in a 13 year-old patient. AB - We present the unusual case of a traumatic "floating patella" in a 13-year-old healthy boy. No predisposing factors were diagnosed. The obvious bony avulsion was treated by immediate open reduction and screwing, whereas the quadriceps tendon rupture was missed diagnostically in the first step. During the course of the treatment, the extension lag lead to the delayed diagnosis of the concomitant rupture of the quadriceps tendon. After the surgical treatment of the latter, the patient healed with full function of the extensor apparatus and full ROM. Bifocal injury of the knee extensor apparatus is therefore possible in young adolescents without presenting the predisposing factors and should thus be considered. PMID- 16217672 TI - The treatment of proximal humeral fractures with intramedullary titanium helix wire by 97 patients. AB - INTRODUCTION: The displaced proximal humeral fracture continues to be a problem and remains the "unsolved fracture". Many guidelines for surgical treatment of different types of these fractures have been described. A conservative functional therapy is recommended as well as a closed or open reposition and internal fixation. Other authors prefer the primary prosthetic replacement of the displaced humeral head. The purpose of this article is to describe a minimal invasive technique--the intramedullary titanium helix wire--and to analyse results and specific problems of this method compared to other techniques. MATERIALS AND METHODS: In a prospective study done over a period of 5 years from 1998 to 2002, 97 patients with two-, three- and four-part-fractures of the proximal humerus were treated with the intramedullary titanium helix wire and given follow-up clinical and radiological checks after 12 months. The functional outcome was evaluated with a 1-year follow-up in 95 of 97 patients. RESULTS: Our 1-year results include: 41 very good, 24 good, 16 satisfactory, 14 poor (average 76 points Constant functional score, 30.7 points UCLA score). CONCLUSION: The advantages of the method arising from the results lie in both the simple operation approach without nerve lesions or wound infections, and also in the indirect, intramedullary splinting of the fracture without damage to the surrounding soft parts, while iatrogenic necroses of the humeral head can be avoided. PMID- 16217673 TI - Association of sporadic Creutzfeldt-Jakob disease with homozygous genotypes at PRNP codons 129 and 219 in the Korean population. AB - Human prion protein gene (PRNP) is considered an important gene in determining the incidence of human transmissible spongiform encephalopathies or prion diseases. Polymorphisms of PRNP at codon 129 in Europeans and codon 219 in Japanese may play an important role in the susceptibility to sporadic Creutzfeldt Jakob disease (CJD); data regarding codon 129 in the Japanese population have led to divergent interpretations. In order to determine which, if any, of the PRNP genotypes in Korean people are associated with sporadic CJD, we examined the genotype and allelic distributions of human PRNP polymorphisms in 150 patients with sporadic CJD. All Korean sporadic CJD patients were Met/Met at codon 129, Glu/Glu at codon 219 and undeleted at the octarepeat region of PRNP. Our study showed significant differences in genotype frequency of PRNP at codon 129 (chi 2=8.8998, P=0.0117) or 219 (chi 2=12.6945, P=0.0004) between sporadic CJD and normal controls. Furthermore, the genotype frequency of the heterozygotes for codons 129 and/or 219 showed a significant difference between the normal population and sporadic CJD patients (chi 2=21.0780, P<0.0001). PMID- 16217674 TI - Arthroscopy and microfracture technique in the treatment of osteochondritis dissecans of the humeral capitellum: report of three adolescent gymnasts. AB - The aim of this paper is to report on three cases of symptomatic osteochondritis dissecans of the humeral capitellum in adolescent gymnasts, two females and one male. In all the cases arthroscopic surgery was performed. During arthroscopy, loose osteochondral fragments were removed, the defect was debrided and microfractures were performed. All the three patients regained the full range of motion of the affected elbow, and returned to the high-level gymnastics within a period of 5 months. At 12 months follow-up, all the three patients remained symptomless and were participating in high-level gymnastics. A combination of arthroscopy and the microfracture technique is a reliable method with excellent short-term results in the treatment of the osteochondritis dissecans of the elbow. PMID- 16217675 TI - Dioxin effects on neonatal and infant thyroid function: routes of perinatal exposure, mechanisms of action and evidence from epidemiology studies. AB - OBJECTIVES: Animal experiments suggest that thyroid function alterations in newborns and infants may represent one of the most sensitive markers of toxicity from 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Dioxin can be transferred from the mother to the offspring either in utero or through lactation. It has been suggested that thyroid-hormone alterations produced by dioxin in utero or shortly after birth may underlie long-term effects, such as cognitive-ability and neurodevelopment impairment. In the present review article, we appraise available evidence on the effects of perinatal exposure to dioxin on fetal and infant thyroid function. METHODS: We summarized the routes of perinatal dioxin exposure and research results on possible mechanisms of dioxin toxic effects on thyroid function. We performed a systematic review of epidemiology studies conducted on mother-child pairs exposed to background environmental levels to investigate dioxin effects on neonatal and infant thyroid function. RESULTS: Toxicological and mechanistic data indicate that dioxin may impair thyroid function in exposed newborns and infants. Investigations on background-exposed children have not consistently demonstrated an association between perinatal TCDD exposure and thyroid function, although some of the studies suggest that sub-clinical hypothyroidism may be induced by perinatal dioxin exposure within 3 months from birth. Between studies inconsistencies may be related to lab method differences, mixed exposures, and small sample size of the populations evaluated. CONCLUSION: Epidemiology studies have as yet failed to demonstrate an association between perinatal TCDD exposure and thyroid function alterations in human subjects, although suggestive evidence from animal and in-vitro experimental data is available. PMID- 16217676 TI - The effects of long-term occupational exposure to dust from herbs. AB - INTRODUCTION: Herbs are a heterogeneous group of many species with several thousand plants, which are used in large quantities in the pharmaceutical and food industries. The aim of the study was to analyse the health effects of long term occupational exposure to dust from herbs. METHODS: A group of 150 people occupationally exposed to dust from herbs, consisting of farmers and workers from herbs processing industry, was examined. As a reference group, 50 urban dwellers not exposed to any kind of organic dust were examined. Examined people were interviewed with the help of the ATS questionnaire compiled by Ferris and by the questionnaire developed in the Institute of Agricultural Medicine in Lublin, Poland for examination of work-related symptoms caused by organic dust. The lung function examination (vital capacity (VC), forced expiratory volume in the first second (FEV(1)), and FEV(1)/VC (%) of normal ranges) and allergological tests (skin prick test, precipitin test and inhibition of leukocyte migration (MIF) test) with microbial antigens were conducted. RESULTS: 71.3% (95% CI 64.1-78.6%) of the exposed people reported occurrence of work-related symptoms. A post-shift decrease of spirometric values (VC, VC%) was observed in the exposed group (mean decrease 2.6%, P<0.01). A significant relationship was found between the number of work-related symptoms and decrease of FEV(1) values, both before (Spearman correlation coefficient r=-0.21, P<0.05) and after work (r=-0.31, P<0.01). In allergological tests, the frequencies of positive reactions in the exposed group were significantly higher than in the reference group. Precipitins specific to Pantoea agglomerans were found in 30.6% (95% CI 23.2-38.1%) of exposed, compared to 12.0% among unexposed (P=0.01). The frequency of positive results in the migration inhibition test was significantly higher among exposed workers for all antigens tested. CONCLUSION: Long-term exposure to dust from herbs causes work related symptoms and decrease of lung function parameters, which, finally, may lead to occupational disease. PMID- 16217677 TI - Lead poisoning from souvenir earthenware. AB - A case of massive lead poisoning from juice contained in a Greek earthenware jug as well as six satellite cases of high lead exposure of similar origin is reported. The intoxicated patient was successfully treated with dimercaptosuccinic acid. Ceramic producers should adhere to the longstanding European legislation. PMID- 16217678 TI - Foliar pH as a new plant trait: can it explain variation in foliar chemistry and carbon cycling processes among subarctic plant species and types? AB - Plant traits have become popular as predictors of interspecific variation in important ecosystem properties and processes. Here we introduce foliar pH as a possible new plant trait, and tested whether (1) green leaf pH or leaf litter pH correlates with biochemical and structural foliar traits that are linked to biogeochemical cycling; (2) there is consistent variation in green leaf pH or leaf litter pH among plant types as defined by nutrient uptake mode and higher taxonomy; (3) green leaf pH can predict a significant proportion of variation in leaf digestibility among plant species and types; (4) leaf litter pH can predict a significant proportion of variation in leaf litter decomposability among plant species and types. We found some evidence in support of all four hypotheses for a wide range of species in a subarctic flora, although cryptogams (fern allies and a moss) tended to weaken the patterns by showing relatively poor leaf digestibility or litter decomposability at a given pH. Among seed plant species, green leaf pH itself explained only up to a third of the interspecific variation in leaf digestibility and leaf litter up to a quarter of the interspecific variation in leaf litter decomposability. However, foliar pH substantially improved the power of foliar lignin and/or cellulose concentrations as predictors of these processes when added to regression models as a second variable. When species were aggregated into plant types as defined by higher taxonomy and nutrient uptake mode, green-specific leaf area was a more powerful predictor of digestibility or decomposability than any of the biochemical traits including pH. The usefulness of foliar pH as a new predictive trait, whether or not in combination with other traits, remains to be tested across more plant species, types and biomes, and also in relation to other plant or ecosystem traits and processes. PMID- 16217679 TI - Defining herbivore assemblages in the Kruger National Park: a correlative coherence approach. AB - Spatial associations of seven herbivore species in the Kruger National Park, South Africa, are analyzed using a new technique, Correlative Coherence Analysis (CoCA). CoCA is a generalization of the concept of correlation to more than two sequences of numbers. Prior information on the feeding ecology and metabolic requirements of these species is used to contrast spatial scales at which hypothesized guild aggregation or competition occurs. These hypotheses are tested using 13 years of aerial census data collected during the dry season. Our results are consistent with the hypothesis that distributions of large and small species of the same feeding type (i.e., grazers and browsers) overlap in potentially resource-rich areas, but have lower similarity values across all areas because the higher tolerance of large species for low quality foods results in a more even spatial distribution of large species compared to small species. PMID- 16217680 TI - Incubation period and immune function: a comparative field study among coexisting birds. AB - Developmental periods are integral components of life history strategies that can have important fitness consequences and vary enormously among organisms. However, the selection pressures and mechanisms causing variation in length of developmental periods are poorly understood. Particularly puzzling are prolonged developmental periods, because their selective advantage is unclear. Here we tested the hypotheses that immune function is stronger in species that are attacked at a higher rate by parasites and that prolonged embryonic development allows the development of this stronger immune system. Through a comparative field study among 12 coexisting passerine bird species, we show that species with higher blood parasite prevalence mounted stronger cellular immune responses than species with lower prevalence. These results provide support for the hypothesis that species facing greater selection pressure from parasites invest more in immune function. However, species with longer incubation periods mounted weaker cellular immune responses than species with shorter periods. Therefore, cellular immune responses do not support the hypothesis that longer development time enhances immunocompentence. Future studies should assess other components of the immune system and test alternative causes of variation in incubation periods among bird species. PMID- 16217683 TI - Loss of Fhit expression is associated with poorer survival in gastric cancer but is not an independent prognostic marker. AB - Several studies have reported conflicting results regarding correlations of the loss of Fhit expression with clinicopathological parameters in gastric cancer. We investigated the immunohistochemical expression of Fhit in 362 cases of sporadic advanced gastric adenocarcinoma. The series included 64 cases with microsatellite instability associated with defective mismatch repair genes. Fhit expression resulted absent in 72% of the tumors analyzed. Absence of Fhit expression was more frequent in cases with diffuse and mixed histotype compared to the intestinal histotype (P=0.009). Absence of Fhit expression also correlated with tumor stage (P<0.001), lymph node involvement (P<0.001), presence of distant metastasis (P=0.033), and increasing histological grade (P=0.005). Retained Fhit expression also correlated with microsatellite instability as 61% of instable tumors had lost Fhit expression compared to 74% of microsatellite stable cancers (P=0.050). While loss of Fhit correlates with poorer survival in univariate analysis, it is not an independent prognostic factor in multivariate analysis and is thus not of clinical utility. PMID- 16217684 TI - Expression of sarcotoxin IA gene via a root-specific tob promoter enhanced host resistance against parasitic weeds in tomato plants. AB - We have developed a simple genetic engineering strategy for conferring resistance against parasitic weeds on host plants. Transgenic tomato plants expressing the sarcotoxin IA gene were grown either in polyethylene bags (PE) or in pots inoculated with Orobanche aegyptiaca seeds. The results indicate that transgenic plants exhibited strong inhibition of parasite growth and significantly increased yield as compared with non-transgenic ones. In both PE and pot systems most of the parasite tubercles attached to the transgenic root plants turned necrotic and developed abnormally. Integration and expression of the gene were confirmed by Southern blot, RT-PCR and Western blot analysis. Our results indicate that the insect gene produced in the plant cells was selectively toxic to the parasite and non-toxic to the host plant. PMID- 16217685 TI - Behavior of Hylamer polyethylene in hip arthroplasty: comparison of two gamma sterilization techniques. AB - Hylamer polyethylene was introduced in the 1990s as an alternative to conventional polyethylene. Its chemical and physical properties, and especially its high crystallinity, were claimed to improve resistance to wear. Initially Hylamer devices were sterilized by gamma radiation in air, then the technique was changed and gamma radiation was performed in the absence of oxygen. Clinical experience has shown the early loosening of some devices made from Hylamer. The aim of this study was to ascertain whether early wear and consequent osteolysis was linked to the sterilization method. We retrospectively compared 31 patients with hip prostheses with Hylamer liners sterilized by gamma radiation in air (group 1) with 30 patients with the same prosthesis, but sterilized in the absence of oxygen (group 2). The groups were similar for sex, age, disease, head diameter and material. Mean follow-up was 84 months and no clinical signs of failure were present. Radiographic measurement revealed that wear of group 1 was significantly greater than that of group 2 (0.23 mm/year vs 0.09 mm/year, p=0.001). Periacetabular and femoral osteolysis in group 1 was significantly greater than group 2. In conclusion, the method of sterilization might have influenced the outcome of Hylamer polyethylene liners. PMID- 16217686 TI - [Epidemiological study of prevalence, incidence and neuropathic pain characterization in neurology units. PREVADOL study]. AB - INTRODUCTION: Neuropathic pain is defined as a pain initiated or caused by a lesion or dysfunction in the nervous system. The objectives of the study were to estimate the prevalence and incidence of neuropathic pain in hospital neurology units and primary care centres, to characterize the clinical profile of the patient with neuropathic pain and to know the most frequent treatments in the pharmacological management of this type of pain. METHODS: Observational, cross sectional epidemiological survey carried out in 36 Neurology Units of the national territory (24 primary care centres and 12 hospitals). During 20 consecutive days neurologists collected the diagnoses of all the attended patients by any reason, up to 30 patients/day. In parallel the 20 first consecutive patients with neuropathic pain were chosen for their characterization in depth by means of a specific questionnaire. RESULTS: A total of 12,688 patients were attended and a total of 13,555 diagnoses were collected through 713 consultation days. The most frequent diagnosis was migraine/cephalea, with a prevalence of 23.40% (95% CI: 22.66%-24.14%). Neuropathic pain represented the eighth more frequent diagnosis, with a prevalence in neurology units of 3.88% (95% CI: 3.54%- 4.22%). The prevalence of neuropathic pain was 2.92% in primary care centres and 6.09% in hospital units (p < 0.01). The daily incidence of new neuropathic pain cases was 1.24% (95% CI: 1.05%-1.53%); 1.14% in primary care neurology centres and 1.45% in hospital units. CONCLUSIONS: The data obtained indicate that neuropathic pain is the eighth more frequent diagnosis in the neurology units. Medical assistance request by neuropathic pain is higher in the hospital units. PMID- 16217687 TI - [Memory loss: a reason for consultation]. AB - INTRODUCTION: Memory loss is an increasingly frequent reason for consultation in neurology. The aim of this work is to know the current frequency as well as the characteristics and disorders of the patients who come for this reason. METHODS: We studied 200 patients who came to general neurology consultation due to loss of memory. RESULTS: 18.47% of the patients who came for the first time to a general neurology consultation did so due to memory loss, this being subjective loss of memory (SLM) in 39% of the cases and referred loss of memory (RLM) in 61% of the cases. The diagnostic groups to which the patients belonged are, in diminishing order, the following: degenerative primary dementia type Alzheimer's disease, mild cognitive impairment, mixed dementia, pure vascular dementia, depressive pseudodementia, attributable to drugs, secondary to systemic disease, non Alzheimer's type disease primary degenerative dementia, structural reasons, transitory global amnesia and epilepsy. No disease was found in 13% of them, and the generally came due to SLM. CONCLUSIONS: Frequency of memory loss as a reason for consultation continues to growing. Patients studied due to memory loss, in whom no disease is found, are generally those having SLM. In spite of this, SLM is a good predictor of cognitive deterioration. It is important to systematically study of every patient and consults for loss of memory and to investigate the possible use of drugs or toxics that could alter the memory. PMID- 16217688 TI - [Clinical validity of two scales of instrumental activities in Alzheimer's disease]. AB - INTRODUCTION: Activities of daily living (ADL) are a major domain in the clinical assessment of Alzheimer's disease (AD) patients. However, ADL scales have not been sufficiently validated in Spain. METHODS: Patients attending a neurology outpatient clinic were classified according to the global deterioration scale (GDS). Afterwards, an independent evaluator administers two scales of instrumental activities of daily living (IADL): Lawton and Brody's scale of IADL (SIADL) and Pfeffer's functional activities questionnaire (FAQ). The SIADL was scored in the original way (dichotomic) (SIADLd) and in an alternative way (ordinal) (SIADLo). Internal consistency (Cronbach alpha coefficient), test retest reliability (intraclass correlation coefficient), diagnostic validity (sensitivity, specificity, number of patients correctly classified) and influence of different variables (regression analysis) were analyzed for the SIADLd, the SIADLo and the FAQ. RESULTS: Ninety-eight patients were recruited. Internal consistency, reliability and diagnostic validity were good or excellent for the three scales. The SIADLo showed better diagnostic and scale features than the SIADLd, but the FAQ surpassed both in all the studied variables. No scale was able to make a proper distinction between patients with subjective complaints (GDS 2) and patients without complaints (GDS 1). Sex and age influenced the SIADL score, but not the FAQ score. The FAQ reached a sensitivity of 0.95 and a specificity of 0.88 in the screening of dementia. CONCLUSIONS: The SIADL and the FAQ are useful, valid and reliable tools for the clinical assessment of AD patients. Ordinal scoring is more advantageous than dichotomic scoring in the SIADL, but the FAQ is preferable. PMID- 16217689 TI - [Spanish version of the Memory Impairment Screen (MIS): normative data and discriminant validity]. AB - OBJECTIVE: Present data on discriminative and normative validity for the Memory Impairment Screen (MIS) in a cross-sectional validation study within a dementia clinic setting using a Spanish adaptation of the MIS. Assess its usefulness as a screening instrument for memory problems related to primary dementing disorders, foremost Alzheimer's Disease (AD). MATERIAL AND METHODS: 403 subjects, including 188 demented subjects according to DSM-IV criteria (119 of which presented AD according to NINCDS-ADRDA criteria) and 215 controls, over 50 years. The MIS, a four word memory test using a specific encoding technique and scoring on a 0-8 range, formed part of the neuropsychological workup, but it was not used for diagnosis. Sensitivity, specificity and predictive values positive (PPV) and negative (NPV) were calculated for the MIS against the gold-standard of clinical diagnosis by two blinded clinicians. ROC curves for the discrimination between demented subjects and controls, and AD subjects and controls were plotted. RESULTS: Control subjects were significantly younger and better educated than the demented samples (p<0.001). No sex differences could be established. ROC curves demonstrated excellent discriminative validity of the MIS 402 for dementia on a whole (0.944) and even better for AD (0.978). The MIS presents satisfying results regarding sensitivity and specificity as well as PPV for different base rates, but the most effective cut-off score lies at < or = 4 points. For all demented subjects this cut-off represents a sensitivity of 0.74 and specificity of 0.96; for AD results are 0.86 and 0.96. More importantly, PPV values are above 0.65 for assumed base rates of 10% and more. Negative predictive values are almost 1 for all assumed base rates. CONCLUSION: The Spanish version of the MIS reliably differentiates between normal and demented subjects. Nevertheless one has to consider the setting of the study when interpreting the results. The use of the S MIS seems to be justified as first level dementia screening tool. PMID- 16217690 TI - [Neurocysticercosis: up-dating in diagnosis and treatment]. AB - Cysticercosis occurs when humans become intermediate hosts of Taenia solium after ingesting its eggs. It is the most common helminthic infection of the nervous system, and is endemic in Latin America, Asia and Africa, as well as in industrialized nations with a high immigrant influx of people coming from endemic areas. Neurocysticercosis is a pleomorphic disease due to individual differences in the number and location of the parasites within the nervous system as well as to differences in the severity of the host's immune reaction against the parasite. Common clinical manifestations include epilepsy, focal neurological signs, intracranial hypertension, and cognitive decline. Diagnosis require proper interpretation of clinical, neuroimaging, and serologic data, in the correct epidemiological context. Neuroimaging studies show the parasites as well as the changes they induce in the nervous system. Tests developed to detect anticysticercal antibodies in serum or cerebrospinal fluid present problems inherent to lack of specificity or sensibility, and should not be used by themselves to confirm or exclude the diagnosis. Cysticidal drugs (albendazole and praziquantel) have improved the prognosis of this condition. However, some patients have torpid clinical courses despite therapy. Surgery plays an important role in the management of some forms of the disease, particularly hydrocephalus and intraventricular cysts. PMID- 16217691 TI - [Percutaneous transluminal angioplasty of the subclavian artery in Takayasu disease: results of long-term follow-up]. AB - Takayasu arteritis is a chronic inflammatory arteriopathy of unknown etiology affecting the aorta and proximal portion of its main branches. Although it was initially reported in young women of Oriental descent, its current worldwide distribution is known to affect both sexes. In the last decade, percutaneous transluminal angioplasty (PTA) has emerged as a viable alternative in its treatment. However, the percentage of restenosis is more common in Takayasu disease than atherosclerotic lesions (21% vs 10%), probably due to diffuse inflammatory vascular involvement. Since the introduction of stent, this technique has emerged as a viable alternative to treatment of atherosclerotic stenotic lesions, although its efficacy and safety in Takayasu disease is still unclear. Herein, we report our experience in a woman with subclavian steal syndrome in whom Takayasu disease was diagnosed and treated with subclavian artery angioplasty and stent, with a good outcome during four years of follow-up. PMID- 16217692 TI - Miliary tuberculosis involving the central nervous system. AB - A woman with chronic systemic lupus erythematosus (SLE) was treated with prednisone, and developed an acute febrile neuroinfection. The magnetic resonance images (MRI) showed miliary micronodular lesions distributed diffusely within the central nervous system and lungs. Lumbar puncture showed pleocytosis with predominance of polymorphonuclear leukocytes, hypoglycorrhachia, elevated proteins and smears positive for acid-fast bacilli. The diagnosis was confirmed by culture of Mycobacterium tuberculosis in the cerebrospinal fluid and bronchial biopsy tissue. She was treated with ethambutol, rifampin, isoniazid and pyrazinamide. MRI taken one month later showed significant improvement, but leg weakness persisted. Epidemiologic research showed her uncle with cavitary tuberculosis as the source of infection. He was also treated with combined antituberculous chemotherapy. PMID- 16217693 TI - [Intercostal neuralgia in diffuse idiopathic skeletal hyperostosis (DISH)]. PMID- 16217694 TI - [Tumoral Parkinsonism]. PMID- 16217695 TI - [Dementia of Alzheimer type, studies of clinico-functional correlation and working memory]. PMID- 16217696 TI - [Testing for neuropsychological profiles. Arguments for Overcoming a Minimal Standard]. PMID- 16217697 TI - [Mild cognitive impairment and Alzheimer's disease: an investigation of the CERAD NP test battery]. AB - To investigate the psychometric properties of the German version of the CERAD-NP, neuropsychological deficits were compared between 49 patients with mild cognitive impairment (MCI), 80 patients with Alzheimer's disease (AD), 36 with major depression (MD), and 26 elderly controls. All participants were outpatients of the memory clinic of the Section of Geriatric Psychiatry, Heidelberg University. Diagnoses were established based on clinical examination, laboratory testing, neuroimaging, and routine neuropsychological testing according to the criteria of aging-associated cognitive decline (AACD) for MCI, NINCDS-ADRDA for AD, and DSM IV for MD, respectively. All CERAD-NP subtests discriminated between controls and AD patients with the latter showing significantly (p< or = 0.05) lower test scores. The subtests verbal fluency and constructive apraxia differed significantly between mildly and moderately AD, while the subtests assessing declarative (epsisodic) memory performance showed only minor, non-significant differences between the respective groups. The LKB patients took an intermediate position between controls and AD patients with significantly lower scores in verbal fluency and declarative memory performance than the controls. When compared with the AD patients, MCI patients were significantly impaired in all subtests except constructive apraxia. Relative to the controls, the patients with MD showed a decreased episodic memory performance but no evidence suggesting an impairment in other neuropsychological domains. Our results indicate that the CERAD-NP is a psychometric instrument which allows a sensitive discrimination between mild and moderate AD, MCI, MD and healthy controls. However, sensitivity of discrimination between different stages of dementia varies with respect to the different subtest. While the subtest for episodic memory showed floor effects already for mild dementia, subtests for verbal fluency and constructive apraxia were able to discriminate even between more advanced stages of the disease. PMID- 16217698 TI - [Neurological manifestations of HIV-infection in the era of highly active antiretroviral therapy (HAART)]. AB - After the introduction of highly active antiretroviral therapy (HAART) in 1996 the neurological manifestations of human immunodeficiency virus (HIV-1)-infection did not decline in incidence and prevalence like the other complications of immunodeficiency; in contrast, due to the longer survival times of HAART treated HIV-1-positive individuals, prevalence of virus associated neurological disease increased during the last years, as international studies underline. Therefore, clinicians and HIV-therapists should be able to diagnose HIV-1-associated neurological disease even in early stages. This article describes symptoms and signs, neuro-imaging and cerebrospinal fluid findings as well as therapy options in primary HIV-1-associated neurological disease like encephalo- and myelopathy and polyneuropathy. Furthermore, those opportunistic infections, caused by bacteria, viruses other than HIV and parasites emerging with manifest immunodeficiency and remaining to be relevant in the HAART era are presented from diagnostic, differential-diagnostic and therapeutic points of view. An extra paragraph describes the interaction of HAART with neurological/psychiatric standard therapies. PMID- 16217699 TI - [Gender differences in cognitive functions]. AB - Gender differences in neuropsychological functioning of patients with psychiatric disorders have been studied extensively in the last years. The available studies provide conflicting results, which can be attributed to the complexity of variables influencing cognitive sex differences. In this article we review the literature about gender differences in cognitive functions in healthy men and women and discuss the relevance of hormones, socio-cultural factors, educational factors and training on the occurrence of these sex differences. Furthermore we summarize the results from functional MRI experiments, which is a useful tool for noninvasively localizing areas in the brain involved in specific cognitive functions. PMID- 16217700 TI - [Intervening between brain and mind: an ethical analysis of the new possibilities of the neurosciences]. AB - The fast progress in neuroscience opens up unprecedented opportunities of research and intervention. Due to their far-reaching medical, social and anthropological implications they raise new ethical issues regarding their goals and legitimacy. The "neuroethical evaluative matrix", which is presented in this article, provides an ethical orientation in neuroscientific research and clinical interventions. Its practicability is demonstrated for the following central neuroethical fields: neuroscientific research, neural grafting, psychopharmacology, neuroimaging, neuro- and psychosurgery, and allocation in neuroscience. It will turn out that the neuroethical matrix provides a systematic and practical possibility to obtain a normative reference frame in neuroscience and neurology. PMID- 16217701 TI - [Vertigo: differential diagnosis and treatment]. PMID- 16217702 TI - Neurological manifestations in lysosomal storage disorders - from pathology to first therapeutic possibilities. AB - Lysosomal storage disorders (LSDs) represent a large and heterogeneous group of inborn errors of metabolism with a rare incidence for the single disease but a respectable overall incidence of 1 in 7700 live births. Neurological involvement in LSDs is quite common and in the last years knowledge about the pathology and clinical course of LSDs has been rapidly increased. Enormous progress has been made in the treatment of LSDs by enzyme replacement, substrate reduction and research on gene therapy. This review aims to describe the progress made as well as the present limitations in this particular field of metabolic medicine. It focuses on those storage disorders with major neurological symptoms or complications where treatment is already available (Gaucher disease, Fabry disease, mucopolysaccharidosis type I) or predictable (Pompe disease, MPS II, MPS IV, MPS VI). PMID- 16217703 TI - Brain MRI and proton MRS findings in infants and children with respiratory chain defects. AB - OBJECTIVE: The aim of this study is to describe neuroimaging patterns in children with respiratory chain (RC) defects using magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (MRS) and to assess their role in the diagnostic evaluation. METHOD: Neuroimaging studies of 49 children (newborn to 15 years old) with biochemical evidence of RC defect were reviewed. Patients were divided in 3 groups ("definite" = 24, "probable" = 14, "possible" = 11) according to Modified Adult Criteria for the diagnosis of RC defect. Eighty-one MRI studies were reviewed for deep gray and white matter changes, degree of myelination, cerebral and cerebellar atrophy, and 67 proton MRS studies were assessed for the presence or absence of lactate elevation, as well as NAA/Cr ratio. The findings were compared among the 3 groups with chi-square test. RESULTS: All patients with "pure" myopathy had normal imaging studies. In patients with CNS involvement, significant differences in the frequency of imaging abnormalities among groups were found for deep gray matter (43 %/8 %/0 %; p = 0.01) and for the presence of lactate elevation on proton MRS (81 %/31 %/0 %; p = 0.001). CONCLUSION: Brain MRI and proton MRS abnormalities were observed only in association with clinical CNS involvement. Deep gray matter signal abnormalities on structural imaging and lactate elevation on proton MRS were more frequently observed in the "definite" group and represent neuroimaging markers for RC mitochondriopathy. PMID- 16217704 TI - Seizure control and acceptance of the ketogenic diet in GLUT1 deficiency syndrome: a 2- to 5-year follow-up of 15 children enrolled prospectively. AB - BACKGROUND: GLUT1 deficiency syndrome is caused by impaired glucose transport into the brain resulting in an epileptic encephalopathy, developmental delay, and a complex motor disorder. A ketogenic diet provides an alternative fuel to the brain and effectively restores brain energy metabolism. METHODS: Fifteen children with GLUT1 deficiency syndrome were enrolled prospectively for a 2.0 - 5.5-year follow-up of the effectiveness of a 3 : 1 LCT ketogenic diet. Eight patients enrolled were described previously, seven patients were novel. RESULTS: Four novel heterozygous GLUT1 mutations were identified. 10/15 patients remained seizure-free on the ketogenic diet in monotherapy. In 2/15 patients seizures recurred after 2(1/2) years despite adequate ketosis, but were controlled by add on ethosuximide. In one patient seizures were reduced without complete seizure control. No serious adverse effects occurred and parental satisfaction with the diet was good. 2/15 patients discontinued the diet. CONCLUSION: GLUT1 deficiency syndrome represents a complex childhood encephalopathy that can be treated effectively by means of a ketogenic diet. The response to the diet did not correlate to clinical, biochemical, or genetic features of the disease. In contrast to previous reports, our results indicate that epilepsy is not always completely controlled by a ketogenic diet and can recur in a subset of patients. PMID- 16217705 TI - Phenotypic heterogeneity in two unrelated Danon patients associated with the same LAMP-2 gene mutation. AB - Danon disease, an X-linked cardioskeletal myopathy caused by primary deficiency of lysosome-associated membrane protein-2 (LAMP-2), is clinically characterized by cardiomyopathy, myopathy, and variable mental retardation. The pathological hallmark of the disease is the absence of LAMP-2 immunohistochemical staining in muscle. The LAMP-2 gene mutations reported thus far are generally private mutations. We describe two cases of Danon disease with different clinical presentation, in whom we identified the same exon skipping mutation c.928G>A in the LAMP-2 gene. The first patient was affected by an early onset myopathy and hypertrophic cardiomyopathy (HCM) that partially improved with drug treatment. A first muscle biopsy at age 4 months showed markedly increased glycogen, and acid maltase deficiency was ruled out biochemically. A second muscle biopsy, performed at age 3(1/2) years, showed very mild abnormalities. The second child at age 15 years had mild, diffuse muscle weakness and wasting, moderate mental deficiency, and HCM. Two serial biopsies performed at age 8 and 15 years showed similar findings of multiple esterase-positive vacuoles in type I myofibers. In both patients the immunohistochemical study demonstrated the absence of LAMP-2 in skeletal muscle. PMID- 16217706 TI - Two new mutations in the MTATP6 gene associated with Leigh syndrome. AB - In this study we have analyzed the mtDNA encoded ATPase 6 and 8 genes ( MTATP6 and MTATP8) in two children with Leigh syndrome (LS) and reduced Mg (2+) ATPase activity in muscle mitochondria. In patient 1, with a mild and reversible phenotype, mutational analysis revealed a heteroplasmic T --> C mutation at nt position 9185 (T9185C) in the MTATP6. The mutation resulted in substitution of a highly conserved leucine to proline at codon 220. The proportion of the mutation was > 97 % in the patient's blood and muscle and 85 % in blood of his asymptomatic mother. Patient 2, with severe clinical phenotype and death at 2 years of age, exhibited a novel heteroplasmic T9191C missense mutation in the MTATP6, which converted a highly conserved leucine to a proline at position 222 of the polypeptide. The proportion of the mutation was 90 % in fibroblasts and 94 % muscle tissue. This mutation was absent in the patient's parents and sister suggesting that the mutation was de novo. Our findings expand the spectrum of mutations causing LS and emphasize the role of MTATP6 gene mutations in pathogenesis of LS. PMID- 16217707 TI - Increased levels of GFAP in the cerebrospinal fluid in three subtypes of genetically confirmed Alexander disease. AB - GFAP levels in the CSF were highly elevated in three genetically confirmed cases of Alexander disease clinically conforming with infantile, early and late juvenile forms. No other CSF abnormalities were detected. Assay of CSF-GFAP may prove to be a rapid and cost-effective screening test in clinical variants of Alexander disease and an indicator of GFAP gene mutations. PMID- 16217708 TI - An unexpected cause of a recurrent cerebral hemorrhage. AB - A 4-year-old previously healthy boy presented with a non-traumatic right parietal hemorrhage. A second life-threatening left cerebral hemorrhage occurred three weeks later and was decompressed with a craniotomy. Transthoracic echocardiography revealed a hypermobile elongated tumor of the mitral valve. The cardiac tumor was successfully resected three weeks after the craniotomy. Histological examination of the cardiac tumor revealed a papillary lesion of spindle cells with smooth muscle cell differentiation. In view of the histological findings and the clinical symptoms, a cellular myofibroblastic tumor was considered the most likely diagnosis in our patient. Although a cardiac tumor is a rare cause of a cerebral hemorrhage, a cardiac evaluation is recommended in pediatric patients with a cerebral hemorrhage of unknown etiology. PMID- 16217709 TI - Marchiafava-Bignami-like injury of the corpus callosum in an infant. AB - We report on a 16-month-old boy who presented with truncal ataxia and intermittent nystagmus. Magnetic resonance imaging (MRI) at 19 months showed a T (2)-hyperintensity of the splenium and the genu of the corpus callosum with extension into the adjacent frontal white matter. Diffusion tensor imaging (DTI) revealed a corresponding area of restricted diffusion, suggesting cytotoxic oedema. The extent and localisation of the signal abnormalities mimic tissue injury as seen in Marchiafava-Bignami disease (MBD). Metabolic investigations were normal. Follow-up imaging at 24 months showed a similar T (2)-hyperintensity of the corpus callosum and white matter while on DTI the cytotoxic oedema had resolved. Clinically a remaining truncal and gait ataxia, clumsiness and a developmental delay is seen. Goal of this case report is (a) to present a rare case of Marchiafava-Bignami-like injury of the corpus callosum in an infant and (b) to discuss the neuroradiological imaging findings including MRI and DTI. PMID- 16217710 TI - Late-onset nephrotic syndrome and severe cerebellar atrophy in Galloway-Mowat syndrome. AB - Galloway-Mowat syndrome (GMS) is a rare autosomal-recessive disorder characterised by nephrotic syndrome, microcephaly, and variable brain anomalies. The prognosis is poor with death almost inevitably supervening before the age of 6 years, but atypical cases with later onset of proteinuria and a more protracted course are on record. We report a female offspring from consanguineous parents suffering from microcephaly, profound psychomotor retardation, epilepsy, hiatal hernia, and striking cerebellar atrophy in whom a nephrotic syndrome became apparent at age 16 years. Renal biopsy revealed focal segmental glomerulosclerosis and glomerular basement membrane abnormalities. We postulate that this patient had a milder form of GMS with severe and diffuse cerebellar atrophy as the leading central nervous system abnormality. PMID- 16217711 TI - Atypical MRI findings in Canavan disease: a patient with a mild course. AB - Canavan disease is a severe, progressive leukodystrophy with an autosomal recessive inheritance, caused by aspartoacylase (ASPA) deficiency. The characteristic MRI features include diffuse, symmetrical white matter degeneration in the subcortical areas, with bilateral involvement of the globus pallidus. Proton magnetic resonance spectroscopy of the brain shows an increase in the concentration of N-acetylaspartic acid (NAA). The altered NAA metabolism has been traced to mutations in the gene encoding ASPA, located on chromosome 17 (17p13-ter). We present here a patient with a mild form of Canavan disease confirmed with the absent ASPA activity, atypical MRI findings, related to compound heterozygosity for a missense mutation, p.Tyr288Cys, and the known pan European mutation, the p.Ala305Glu. PMID- 16217712 TI - [Income reduction due to sickness benefits--when does sickness make you poor?]. AB - PURPOSE: When absent from work due to sickness, most employees in Germany receive continued pay from their employer for six weeks. After this period, sick employees receive sickness benefits from their Statutory Sickness Fund. These sickness benefits are calculated in a rather complicated way as a percentage of gross and net salary. The paper focuses on two questions that have rarely been studied: which income groups show a particularly large difference between net salary and net sickness benefits? Which income groups move below the poverty line after receiving sickness benefits? METHODS: We calculated how much sickness benefit is actually paid to the insured, for different income and tax groups. The definition for the poverty line is outlined as well. Due to methodological difficulties, the comparison between sickness benefits and poverty must be confined to single-person households. RESULTS: In the income groups chosen here (gross salary up to 4000 Euro per month), net sickness benefits amount to about 77 % of net salary, for all insured. Financial problems can mainly be expected for the lower and the upper income groups. Expressed in absolute terms, the upper income groups experience a large reduction in net income. The lower income groups come close to the poverty line or fall below it. CONCLUSIONS: Sickness benefits provide income in case of sickness; this is an important achievement of social policy. However, we should study the financial burden which sickness benefits could have for the insured. More in-depth analyses would require data that are not yet available (e. g. on the number of insured per income group and the income of other household members). The analyses presented here already show that sickness benefits could lead to severe financial problems for at least some insured. They point to the need for more studies in this neglected field. PMID- 16217713 TI - [The new private health insurance subscribers]. AB - The new German legislation concerning the modernisation of statutory health insurance allows statutory health insurers to cooperate with their private health counterparts to offer supplementary health insurance. This study investigates the demand for such policies and how much someone is prepared to pay for them. Furthermore, the study tries to characterise the subscribers of supplementary health insurance. This analysis is based on data from the TNS Healthcare Access Panel on 896 statutory health insured persons aged 20 to 79 years. Statutory health insured persons show a great interest in supplementary health insurance policies. 67.3 % are prepared to subscribe to one or more policies and pay monthly 29.37 on average. Subscribers to supplementary health insurance policies also support a model of statutory health insurance which gives them an opportunity to choose between a basic level of cover and the extended benefits of a supplementary health insurance policy. In addition, they are high earners and rarely see a physician. They are very content with their statutory health insurers; however, their opinion about the German health system differs because part of the subscribers are very content and others are dissatisfied. Moreover, the study shows that persons who see a physician often and who assess their state of health poorly would significantly buy fewer supplementary health insurance policies. It is not certain why this group come to such a decision. However, if supplementary insurance policies help to determine the levels of access granted to see a physician then this study demonstrates that persons with a high demand for medical care and poor persons are excluded. PMID- 16217714 TI - [Reproductive medicine in Germany--peculiarities and dilemmas]. AB - BACKGROUND: Technological progress, as well as increasing success, of reproductive medicine may lower the threshold for childless couples to initiate infertility therapy. However this development may aggravate unsolved problems (e. g. multiple births) and may not increase the pregnancy rate, due to the present unsatisfactory legal situation in Germany (e. g. ban on selective blastocyst culture). METHODS: Based on a systematic review of the literature, we studied actual problems of German reproductive medicine under three public health-related topics: (1) the decline in population in Germany, (2) criteria for success in reproductive medicine and (3) new treatment options in the light of legal and ethical aspects. RESULTS: In Germany (and other industrial nations), reproductive medicine has emerged parallel to an ever-increasing rate of childlessness. Today, 2 % of all births result from treatments using the techniques of reproductive medicine. Nevertheless, the actual extent of involuntary childlessness is lower (below 8 %) than commonly suggested (between 10 and 15 %). Frequently, the success of treatment depends upon a series of treatment cycles and is, especially in Germany, accompanied by a high rate of multiple births (more than 30 %). Hence, a more adequate success rate may be the healthy, i. e., term, singleton baby. The use of selective blastocyst culture, currently not permitted in Germany, could further improve therapy or, at least, reduce patient stress and discomfort. CONCLUSION: Reproductive medicine, as seen from a public health perspective, needs to pursue patient-oriented requirements more in-depth. An open discussion of new technologies which could improve reproductive health but are at present not permitted, would be imperative. PMID- 16217715 TI - [Accessibility of doctors' surgeries in Essen, Germany]. AB - BACKGROUND: Constructional barriers often prevent persons who are only partially able, for example those requiring a wheel chair for pre-ambulation, from entering buildings where doctors practise. Even though many international and national resolutions have long been demanding free access to the environment for the partially able, this has not been specially prescribed in Germany. Hence, no one knows anything about outpatient health care facilities in this regard. The present study aimed at analysing accessibility to orthopaedic and neurological practices and surgeries for wheelchair patients. METHOD: We chose Essen, the sixth largest town in Germany, as an example of an urban area, where orthopaedists and neurologists are frequently accessed by wheelchair patients. We performed on-site investigations of the exterior and interior zones of all orthopaedic and neurological surgery buildings in Essen (each n = 29). Criteria for our descriptive analysis were parking lots for the handicapped, shunting areas, entrances at-grade, steps/stories, banisters, ramps, bells and openers of front, elevator and surgery doors, their opening and width. Following the criteria of the DIN 18 024 standard part 2 ("accessibility") the surgeries were divided into four groups 1) fully accessible; 2) slight barriers; 3) considerable barriers; 4) massive barriers. RESULTS: None of the 58 investigated surgeries was fully accessible, 21 of the 29 surgeries of each medical specialty had massive barriers, so that wheelchair patients could access these surgeries only with the help of at least two (strong) persons. Six of the 29 orthopaedic and three of the 29 neurological surgeries had slight barriers, whereas two orthopaedic and five neurological surgeries had distinct barriers. Main barriers were steps in the entrance area; front, elevator or surgery doors too narrow (width less than 80 cm), and elevators too small. DISCUSSION: For wheelchair patients in Germany, free choice of doctors seems to be massively reduced. Since 80 % of orthopaedic and 90 % of neurological surgeries in Essen do not fulfil the quality feature "constructional accessibility", measures that have been taken in the past to help partially able persons to participate in this respective aspect of social life have not been effective. New measures to improve the present situation should be agreed upon by all the institutions involved (politics, local authorities/construction supervision, sickness funds, doctors and associations of sickness fund physicians, and concerned persons). If voluntary measures do not lead to free choice of doctors for wheelchair patients, further legal regulations appear to be mandatory. PMID- 16217716 TI - [Health related self-help groups--participation and information level in Germany. Results of the German telephone health survey 2003]. AB - Using the data provided by the German Telephone Health Survey 2003 that gathered health related information from 8,318 adult residents, statements concerning the participation in health related self-help groups and the level of information about self-help groups were analysed. According to the survey 8.9 % of sample have already participated in a health related self-help group, 13 % stated to be informed about this topic but have not taken part in such a group. Women are more often informed about self-help groups and have more frequently reported on participation experience than men. In comparison the level of information and participation in former East Germany is slightly smaller than in West Germany. Further, the frequency of participation experience reported as well as the level of information increased with socioeconomic status; the differences between the social stratums were smaller for participation level than for information level. People reporting to suffer from chronic health conditions or to have severe handicaps, stated approximately twice as often to be informed about self-help groups as people who did not so. Compared to estimates for the beginning of the eighties the results indicate an increased part of German population stating experiences with health related self-help groups. Especially persons affected by chronic illnesses and severe handicaps reported to be or have been active in self help groups. However, apparently a considerable unused self-help potential still exists. PMID- 16217717 TI - [Smoking and low socio-economic status]. AB - People of lower socio-economic strata increasingly use legal as well as illegal drugs. Tobacco and alcohol are legal drugs that cause particular concern. Both drugs are widely abused in Germany by people attempting to escape their everyday problems. For decades it has been known that tobacco and alcohol use are more prevalent in lower socio-economic strata of society (those with low educational achievement, compared to people with further or higher education qualifications). Tobacco and alcohol abuse is particularly high among the unemployed, either temporarily or longterm, as well as among persons living alone. Children and women are more concerned about smoking than men. Male loneliness, often accompanied by the appearance of depressive reactions or of depression, increases the likelihood of cigarette smoking. Poor people spend up to 20 % of their income on tobacco. In many industrialised countries, the age of onset of smoking is becoming younger and younger, increasing the risk of development of avoidable tobacco-related illnesses at an earlier age. This means that young smokers who develop chronic tobacco-related illnesses require medical care for many years, increasing the cost of treating tobacco-related disease. Within the next few years, effective prevention programmes against smoking must be developed, particularly for the lower socio-economic populations, to prevent the cost of health care systems spiralling during the next few decades. PMID- 16217718 TI - [Sample survey of persons insured in statutory health insurance institutions in Hessen--concept and realisation of person-related data base]. AB - Statutory health insurance data are being increasingly used for secondary data research. Longitudinal data can be prepared for research in health care, epidemiology or demand planning, in particular through the person-related nature of the data which is a precondition for the creation of inter-sector and inter period data sets. This application possibility was introduced in a method study "person-related sampling of statutory health insurance data" and is now translated into practice on a larger scale for the first time in the regional sample "Versichertenstichprobe AOK Hessen/KV Hessen". For the collection and use of these data, model procedures were designed which take account of organisational (data access, contractual agreement, advisory board), technical (sampling, collection and storage of data) and confidentiality (data protection concept, pseudonymisation) aspects. The insured person-related sample may thus serve as a basis for the data pool planned for the national health system (Social Security Regulation 303 a-SGB V). PMID- 16217719 TI - [Effects of interviews during body weight checks in general population surveys]. AB - While surveying actually measured body weight is largely impractical in national surveys, self-reported weight is a simple and inexpensive method of collecting data. Previous research shows that data on reported body weight are falsified by systematic mis-reporting. This bias is said to be the consequence of the sensitive nature of information on body weight. Numerous studies on survey response suggest that certain modes of data collection are more conducive than others for probing sensitive information. This paper investigates the effect of the anonymous interviews, characteristics of the interviewer and respondents' familiarity with the survey, as factors that may impinge on reported body weight. Findings of this paper show that refusals to state the body weight are rare. Moreover, characteristics of interviewers account for only a small fraction of the variance in reported body weight. Yet the hypothesis that the absence of an interviewer in self-administered interviews increases reported body weight can be confirmed. This interview effect, however, occurred in men only. On average, male respondents in anonymous interview settings report on a body weight which is 1 kg more than they would report in other settings. The repeated participation of respondents in the Socio-Economic Panel Study (SOEP) increases their reported body weight accuracy which suggests a positive panel effect on respondents' willingness to disclose sensitive information. PMID- 16217720 TI - [Quality of life assessment in Inflammatory Bowel Disease (IBD): German version of the Inflammatory Bowel Disease Questionnaire (IBDQ-D; disease-specific instrument for quality of life assessment) -- first application and comparison with international investigations]. AB - BACKGROUND: Health-related quality of life (HRQOL) is an important outcome parameter in health research and care. The aim of the working group Quality of Life in the Competence Network Inflammatory Bowel Disease (IBD; in the original German: "Kompetenznetz chronisch entzundliche Darmerkrankungen") is to generate instruments for assessment of HRQOL and its implementation as standards in clinical trials, health care and research in IBD. METHODS: The Inflammatory Bowel Disease Questionnaire (IBDQ) is an international validated disease specific instrument for HRQOL-assessment. A German version of the IBDQ was elaborated and tested in 415 outpatients with Crohn's disease (CD, n = 306) and ulcerative colitis (UC, n = 109). The aim of the study was to compare the results of HRQOL assessment (IBDQ-D) with international investigations, to correlate HRQOL results with disease activity and to preform a pretest of psychometric properties. RESULTS: International data suggest that the IBDQ-D is a suitable instrument for HRQOL-assessment in CD and UC. For both disease a statistically significant negative correlation with disease activity was found. Tested psychometric properties do not suggest that a revision of the IBDQ-D is required. The IBDQ-D offers the HRQOL-assessment as an primary or secondary outcome in clinical trials in IBD in Germany. PMID- 16217721 TI - [Speech therapy assessment: field study on psychometric properties, practicability, acceptance and process quality]. AB - OBJECTIVE: to investigate the internal consistency, responsiveness, discriminative validity, practicability, acceptance and process quality of a recently developed Speech Therapy Assessment (STA) under routine work conditions of speech and language therapists in German speaking countries. Since standardised, generic and ICF-oriented assessment tools for documentation and evaluation of speech therapy interventions for adult clients are missing in German speaking countries and existing tests cover only sub-areas, the STA has been developed in the years 1995 to 2002. By means of different domains, speech and language therapists assess client (1) communication, (2) aphasia, (3) speech apraxia, (4) dysarthria and (5) dysphagia as well as (6) her or his dealing with corresponding disabilities. METHODS: 17 therapists from 14 institutions applied the STA to 260 adult clients with language, speaking or swallowing disorders. The clients were included in the study consecutively over a period of 7 month. After this period, the therapists completed a questionnaire regarding the benefit and practicability of the STA. Cronbach alpha was calculated as indicator for internal consistency, effect sizes (standardised response means) for responsiveness and ROC values for discriminative validity. The answers of the questionnaire about the benefit of the STA were evaluated both, quantitatively and qualitatively. RESULTS: The internal consistency and discriminative validity were high (Cronbach alpha: 0.79 to 0.95; ROC-values: 0.84 to 0.98). Effect sizes regarding responsiveness were moderate (standardised response means: 0.46 to 0.78). On a 5-step Likert scale (1 = very good, 5 = inadequate), the therapists rated the average (standard deviation) benefit of the STA with: practicability 2.6 (1.2), acceptance 2.8 (1.3), impact on diagnostics 2.8 (1.3), impact on finding therapeutic goals 3.5 (1.2), impact on communication with other rehabilitation partners 2.7 (1.5) and overall judgement 2.6 (0.9). CONCLUSION: The STA fulfils essential quality criteria of the classical test theory. The involved therapists assessed the benefit of the STA as satisfactory. In addition, they pointed out concrete improvement potential for the implementation in practice. It is planed to investigate a refined version of the STA in a multi centre validation study. PMID- 16217722 TI - [Psychosocial stress at work and musculoskeletal pain among police officers in special forces]. AB - This study investigates associations of an adverse psychosocial work environment with musculoskeletal pain among German police officers in special forces. Analyses are based on a survey of 480 officers in special police forces (mainly special assignments and mobile units). An adverse psychosocial work environment is measured by the effort-reward imbalance model that identifies "high effort/low reward" working conditions. Musculoskeletal pain is assessed by means of a validated questionnaire (12 months and 7 days prevalence). Analyses confirm that police work is a stressful occupation. Multivariate logistic regression analysis indicates a twofold risk for neck, back and hip pain among police officers defined by an imbalance of high effort and low reward at work after adjustment for age, gender, socio-economic status, and physical work load. PMID- 16217723 TI - Mineralization of regenerated cellulose hydrogels induced by human bone marrow stromal cells. AB - The proliferation of cultured human bone marrow stromal cells (HBMSC) on regenerated cellulose hydrogels was assessed. Regenerated cellulose hydrogels showed good rates of HBMSC proliferation, the cells exhibiting a flattened morphology, and after 22 days in culture, the cells had homogeneously colonized the surface of the materials. Moreover, since the early days in culture, between the surface of the materials and attached cells a continuous granulated hydroxyapatite layer was formed. It has been previously demonstrated in vitro, but without cells, that these materials did not mineralize. Hence, it seems that HBMSC promoted the mineralization of the surface. PMID- 16217724 TI - Comparative assessment of three in vitro exposure methods for combustion toxicity. AB - A comparative assessment of three approaches for the use of human cells in vitro to investigate combustion toxicity was conducted. These included one indirect and two direct (passive and dynamic) exposure methods. The indirect method used an impinger system in which culture medium was used to trap the toxicants, whilst the direct exposure involved the use of a Horizontal Harvard Navicyte Chamber at the air/liquid interface. The cytotoxic effects of thermal decomposition products were assessed using the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay (Promega) on a selection of human cells including: HepG2, A549 and skin fibroblasts. A small scale laboratory fire test using a vertical tube furnace was designed for the generation of combustion products. Polymethyl methacrylate (PMMA) was selected as a model polymer to study the cytotoxic effects of combustion products. NOAEC (no observable adverse effect concentration), IC10 (10% inhibitory concentration), IC50 (50% inhibitory concentration) and TLC (total lethal concentration) values were determined from dose response curves. Assessment using the NRU (neutral red uptake) and ATP (adenosine triphosphate) assays on human lung derived cells (A549) was also undertaken. Comparison between in vitro cytotoxicity results against published toxicity data for PMMA combustion and predicted LC50 (50% lethal concentration) values calculated from identified compounds using GCMS (gas chromatography mass spectrometry) was determined. The results suggested that the indirect exposure method did not appear to simulate closely exposure via inhalation, whilst exposure at the air/liquid interface by using the dynamic method proved to be a more representative method of human inhalation. This exposure method may be a potential system for in vitro cytotoxicity testing in combustion toxicity. PMID- 16217725 TI - Effects of kaolin-based particle film on spruce budworm (Choristoneura fumiferana (Lepidoptera: Tortricidae)) oviposition in the laboratory. AB - A laboratory study conducted in 2003 determined the influence of a kaolin-based particle film (Surround WP Crop Protectant) on spruce budworm (Choristoneura fumiferana (Clem)) oviposition. Three concentrations (15, 30 and 60 g litre(-1) spray carrier) were applied to white spruce (Picea glauca (Moench) Voss). When presented with different paired choices of oviposition surfaces, spruce budworms laid very few egg masses overall and showed no significant (P < 0.05) preference between kaolin-coated and untreated foliage. During no-choice tests, spruce budworms were significantly (P < 0.05) less inclined to oviposit on the 60 g kaolin litre(-1) treated foliage than on the controls, but no kaolin treatment completely inhibited spruce budworm oviposition. Egg mass size and percentage hatch were unaffected by the kaolin treatments, and overall the percentage of egg masses laid on foliage was inversely proportional to treatment concentration. It is unlikely that kaolin-based particle film would be practical for wide use in natural forests without significant adaptations to current pesticide application equipment and methods. However, the technique might be feasible in selected urban settings. PMID- 16217726 TI - Metabolic regulation at the tricarboxylic acid and glyoxylate cycles of the lignin-degrading basidiomycete Phanerochaete chrysosporium against exogenous addition of vanillin. AB - A proteomic differential display technique was utilized to study cellular responses of Phanerochaete chrysosporium exposed to vanillin, one of the key intermediates found during lignin biodegradation. Intracellular proteins were resolved by 2-DE and target protein spots were identified using MALDI-MS after in gel tryptic digestions. Upon addition of vanillin to P. chrysosporium, up regulation of homogentisate 1,2-dioxygenase, 1,4-benzoquinone reductases, aldehyde dehydrogenase, and aryl-alcohol dehydrogenase, which seem to play roles in vanillin metabolism, was observed. Furthermore, enzymes involved in glycolysis, the tricarboxylic acid cycle, the pentose-phosphate cycle, and heme biosynthesis were also activated. Up-regulation of extracellular peroxidase was also observed. One of the most unique phenomena against exogenous vanillin was a switch from the glyoxylate cycle to the tricarboxylic acid cycle, where a drastic increase in isocitrate dehydrogenase activity was observed. The exogenous addition of other aromatic compounds also caused an increase in its activity, which in turn triggered NAD(P)H production via the action of dehydrogenases in the tricarboxylic acid cycle, heme biosynthesis via the action of aminolevulinic acid synthase on succinyl-CoA, and energy production via activation of the mitochondrial electron transfer system. These metabolic shifts seem to be required for activating a metabolic system for aromatic compounds. PMID- 16217729 TI - From proteins to systems: the British Society for Proteomics Research (BSPR) meeting 2005. AB - This report describes the highlights of the second scientific meeting of the British Society for Proteome Research (BSPR), jointly organised with the European Bioinformatics Institute (EBI), and held at The Genome Centre, Cambridge UK in July 2005. The theme of the meeting was "From Proteins to Systems" covering many diverse aspects of proteomics, bioinformatics and systems biology. PMID- 16217730 TI - Experience-altered oviposition responses to a neem-based product, Neemix, by the diamondback moth, Plutella xylostella. AB - The oviposition responses of the diamondback moth, Plutella xylostella L., to Neemix 4.5, a neem-based oviposition repellent/deterrent, with or without previous experience were studied in the laboratory on plain aluminum foil sheets, on aluminum foil sheets coated with cabbage juice, and on cabbage plants. In the plain aluminum foil sheet experiment, the females without prior experience of Neemix (inexperienced females) deposited more eggs (66.2%) on sections with untreated sheets than on sections with Neemix-treated sheets (33.8%), indicating that the inexperienced females were significantly repelled and/or deterred by Neemix. In contrast, the Neemix-experienced females deposited more eggs (69.2 69.7%) on sections with Neemix-treated sheets than on sections with untreated sheets (30.3-30.8%), implying that the females were not repelled and/or deterred by Neemix following an experience. In the tests using egg-laying sheets coated with cabbage juice or with cabbage juice plus Neemix, the inexperienced females deposited more eggs (63.8%) on the sections with sheets treated with cabbage juice only than on the sections with sheets coated with cabbage juice and Neemix (36.2%), indicating that inexperienced females were repelled and/or deterred by Neemix. Similarly, Neemix-experienced females deposited more eggs (62.2-65.6%) on the sections with sheets treated with cabbage juice plus Neemix than on the sections with sheets treated with juice only (34.4-37.8%), suggesting that the females were not repelled and/or deterred by Neemix following an experience. On cabbage plants, inexperienced females oviposited similar numbers of eggs on both Neemix-treated (50.8%) and untreated plants (49.2%). However, the Neemix experienced females oviposited more eggs on the plants treated with Neemix (56.3%) than on untreated plants (43.7%), indicating that the females were attracted by Neemix-treated plants following an experience. The significance of this study for applications of insect repellents/deterrents in pest management is discussed. PMID- 16217731 TI - Comparing ecological risks of pesticides: the utility of a Risk Quotient ranking approach across refinements of exposure. AB - Environmental risk assessment of pesticides and other chemicals often uses the Risk Quotient (RQ) method to characterize risk quantitatively. An RQ is calculated by dividing an environmental exposure value by a toxicity end-point value. Tier 1 RQs, which are characterized by highly conservative toxicity and exposure assumptions, are used primarily for screening out negligible risks in regulatory decision making. It has been argued that the tier 1 RQ approach is valuable for making direct comparisons of quantitative risk between pesticides. However, an outstanding question is whether relative risks among pesticides would change if refinements of exposure are incorporated into the RQ calculations. This study tested that hypothesis. Aquatic ecological risk assessments were conducted for 12 herbicide and 12 insecticide active ingredients used on agricultural crops in the USA. The pesticides were chosen because surface-water monitoring data for them were available as part of the United States Geological Survey's National Water-Quality Assessment Program (NAWQA). Ecological receptors and effects evaluated were aquatic non-vascular plants (acute risk), aquatic vertebrates (acute risk) and aquatic invertebrates (acute risk) for the herbicides and aquatic vertebrates (acute and chronic risk) and aquatic invertebrates (acute and chronic risk) for the insecticides. The data indicate that there were significant statistical correlations between numerical rankings of tier 1 RQs and RQs using refined environmental exposures. The results support the hypothesis that numerical ranking of RQs for the purpose of comparing potential ecological risks is a valid approach because the rankings are significantly correlated regardless of the degree of exposure refinement. PMID- 16217732 TI - Masking the taste of the conditioned taste aversion agent levamisole using an ion exchange resin, for practical application in wildlife management. AB - For a conditioned taste aversion (CTA) agent to be successful in wildlife management applications, the compound must not be detectable by the animal. Levamisole is an effective CTA agent when administered by oral intubation, but it is readily detected by a number of species when mixed directly in food. This paper describes the development of an ion-exchange resin complex (resinate) to mask the taste of levamisole. Two different resins were evaluated, Amberlite IRP 64 and Amberlite IRP-69, and release studies indicated that the resinate formed using IRP-64 resin would be most suitable for use in wildlife management. Although it contained a relatively low loading of levamisole (77 g kg(-1)), the results indicated that the IRP-64 resinate should be stable in the mouth and release the levamisole quickly in the acid environment of the stomach (93% of levamisole was released into 0.1 M HCl in 5 min). In a bioassay using laboratory rats (Rattus norvegicus Berk), we showed that the taste of levamisole was successfully masked in a biscuit bait using the IRP-64 resinate and that a CTA was generated to untreated bait. The use of ion-exchange resins is a new approach in the taste-masking of CTA agents and could be applied to other wildlife management applications. PMID- 16217733 TI - Diphenylphosphanylsulfoximines as ligands in iridium-catalyzed asymmetric imine hydrogenations. PMID- 16217734 TI - Isolation of a mRNA encoding a glycine-proline-rich beta-keratin expressed in the regenerating epidermis of lizard. AB - During scale regeneration in lizard tail, an active differentiation of beta keratin synthesizing cells occurs. The cDNA and amino acid sequence of a lizard beta-keratin has been obtained from mRNA isolated from regenerating epidermis. Degenerate oligonucleotides, selected from the translated amino acid sequence of a lizard claw protein, were used to amplify a specific lizard keratin cDNA fragment from the mRNA after reverse transcription with poly dT primer and subsequent polymerase chain reaction (3'-rapid amplification of cDNA ends analysis, 3'-RACE). The new sequence was used to design specific primers to obtain the complete cDNA sequence by 5'-RACE. The 835-nucleotide cDNA sequence encodes a glycine-proline-rich protein containing 163 amino acids with a molecular mass of 15.5 kDa; 4.3% of its amino acids is represented by cysteine, 4.9% by tyrosine, 8.0% by proline, and 29.4% by glycine. Tyrosine is linked to glycine, and proline is present mainly in the central region of the protein. Repeated glycine-glycine-X and glycine-X amino acid sequences are localized near the N-amino and C-terminal regions. The protein has the central amino acid region similar to that of claw-feather, whereas the head and tail regions are similar to glycine-tyrosine-rich proteins of mammalian hairs. In situ hybridization analysis at light and electron microscope reveals that the corresponding mRNA is expressed in cells of the differentiating beta-layers of the regenerating scales. The synthesis of beta-keratin from its mRNA occurs among ribosomes or is associated with the surface of beta-keratin filaments. PMID- 16217735 TI - Tissue interactions in the regulation of axon pathfinding during tooth morphogenesis. AB - Like many other organs, the tooth develops as a result of the epithelial mesenchymal interactions. In addition, the tooth is a well-defined peripheral target organ for sensory trigeminal nerves, which are required for the function and protection of the teeth. Dental trigeminal axon growth and patterning are tightly linked with advancing tooth morphogenesis and cell differentiation. This review summarizes recent findings on the regulation of dental axon pathfinding, which have provided evidence that the development of tooth trigeminal innervation is controlled by epithelial-mesenchymal interactions. The early dental epithelium possesses the information to instruct tooth nerve supply, and signals mediating these interactions are part of the signaling networks regulating tooth morphogenesis. Tissue interactions, thus, appear to provide a central mechanism of spatiotemporally orchestrating tooth formation and dental axon navigation and patterning. PMID- 16217736 TI - A non-canonical photopigment, melanopsin, is expressed in the differentiating ganglion, horizontal, and bipolar cells of the chicken retina. AB - Vertebrate melanopsin is a photopigment in the eye, required for photoentrainment. Melanopsin is more closely related to opsin proteins found in invertebrates, than to the other photo-pigments. Although the invertebrate melanopsin-like protein is localized in rhabdomeric photoreceptors in the invertebrate eye, it has been shown to be expressed in a subset of retinal ganglion cells in the mouse and in horizontal cells in the frog, indicating its diversified expression pattern in vertebrates. Here we show that two types of melanopsin transcripts are expressed in the developing chicken retina. Melanopsin is firstly expressed by a small subset of ganglion cells, and then prominently expressed by horizontal cells and later by bipolar cells in the developing chicken retina. This suggests that a subset of ganglion, horizontal, and bipolar cells in the chicken retina may have rhabdomeric properties in their origins. PMID- 16217738 TI - Expression of synaptic vesicle two-related protein SVOP in the developing nervous system of Xenopus laevis. AB - Synaptic vesicle-associated proteins are important regulators of neurotransmitter release at synaptic terminals in mature animals. Some synaptic vesicle-associated proteins are also expressed during development, although their contribution to development is not as clear. Here, we describe the cloning and developmental expression pattern of the Xenopus laevis synaptic vesicle-associated protein SVOP, a gene first identified as an immediate target for proneural basic helix loop-helix factors. Alignment analysis revealed a high level of identity between the SVOP protein sequences from Xenopus and other vertebrates. In developing Xenopus embryos, SVOP expression is restricted to the nervous system and is first detectable at the mid-neurula stage. As development progresses SVOP becomes broadly expressed throughout the central nervous system. Our observation that SVOP is expressed in the developing Xenopus nervous system suggests that it may be involved in neuron formation, maturation, or neuronal function. PMID- 16217737 TI - Inner ear formation during the early larval development of Xenopus laevis. AB - The formation of the eight independent endorgan compartments (sacculus, utricle, horizontal canal, anterior canal, posterior canal, lagena, amphibian papilla, and basilar papilla) of the Xenopus laevis inner ear is illustrated as the otic vesicle develops into a complex labyrinthine structure. The morphology of transverse sections and whole-mounts of the inner ear was assessed in seven developmental stages (28, 31, 37, 42, 45, 47, 50) using brightfield and laser scanning confocal microscopy. The presence of mechanosensory hair cells in the sensory epithelia was determined by identification of stereociliary bundles in cryosectioned tissue and whole-mounts of the inner ear labeled with the fluorescent F-actin probe Alexa-488 phalloidin. Between stages 28 and 45, the otic vesicle grows in size, stereociliary bundles appear and increase in number, and the pars inferior and pars superior become visible. The initial formation of vestibular compartments with their nascent stereociliary bundles is seen by larval stage 47, and all eight vestibular and auditory compartments with their characteristic sensory fields are present by larval stage 50. Thus, in Xenopus, inner ear compartments are established between stages 45 and 50, a 2-week period during which the ear quadruples in length in the anteroposterior dimension. The anatomical images presented here demonstrate the morphological changes that occur as the otic vesicle forms the auditory and vestibular endorgans of the inner ear. These images provide a resource for investigations of gene expression patterns in Xenopus during inner ear compartmentalization and morphogenesis. PMID- 16217739 TI - Guest editors' introduction: is EMF a potential environmental risk for children? PMID- 16217740 TI - Factors influencing distress in Indian cancer patients. AB - BACKGROUND: The current study set out to identify distress in cancer patients undergoing curative treatment within India. PATIENTS AND METHODS: This study was carried out to measure distress and contributory factors in 103 cancer patients undergoing treatment with curative intent. The patients were interviewed using the Distress Inventory for Cancer (DI-C). The data on social, demographic, clinical, treatment, and follow-up details was collected from case records. RESULTS AND CONCLUSION: The distress score for individual respondents ranged from 34 to 90 (mean 62.3). Patients with lower income, those who were single/widowed, or divorced, those living between 150 and 350 km (3-6 h commuting distance) from the cancer centre, presence of pain and patients with advanced tumours at presentation showed higher distress. A higher distress score correlated significantly with patients being lost to follow-up. PMID- 16217741 TI - Post-embryonic expression of C. elegans microRNAs belonging to the lin-4 and let 7 families in the hypodermis and the reproductive system. AB - MicroRNAs (miRNAs) are regulatory molecules that negatively control gene expression by binding to complementary sequences on target mRNAs. The most thoroughly characterized miRNAs, lin-4 and let-7, direct cell fate determination during the larval transitions in C. elegans and act as key regulators of temporal gene expression. lin-4 and let-7 are founding members of two distinct families of miRNA genes sharing strong sequence homology primarily in the 5' end of the mature miRNAs. In this report, we characterize the temporal and spatial expression patterns of lin-4 and let-7 family members using northern blot analysis and mir::gfp fusion studies. Our results show that lin-4 and let-7 homologues possess distinct temporal and spatial expression patterns during nematode development and that known heterochronic genes regulate their expression. We find that certain lin-4 and let-7 family members display overlapping expression patterns in the hypodermis and the reproductive system, suggesting that combinations of miRNAs from across families may control common developmental events. PMID- 16217742 TI - Retinoic acid signaling is required for proper morphogenesis of mammary gland. AB - Retinoic acid (RA), a bioactive chemical compound synthesized from dietary derived vitamin A, has been successfully used as a chemopreventive and chemotherapeutic agent through the regulation of cell proliferation, differentiation, and apoptosis acting via the retinoic acid receptors. Despite two decades of research on the function of retinoic acid, the physiological role of RA in mammary gland development is still not well characterized. In this report, we demonstrate that RA is required for proper morphogenesis of mouse mammary gland in a novel transgenic mouse model system. It was found that inhibition of RA signaling in vivo leads to excessive mammary ductal morphogenesis through upregulation of cyclin D1 and MMP-3 expression. Furthermore, we show that the transgene-induced excessive branching morphogenesis could be reversed by treatment with RA, demonstrating the direct physiological effect of RA signaling in vivo. In addition, we demonstrate that excessive branching morphogenesis in the transgenic mammary gland are cell-autonomous and do not require stromal signals within the transgenic mammary gland. Finally, we provide evidence suggesting that retinoic acid signaling is required for appropriate mammary gland differentiation. Collectively, our data indicate for the first time that retinoic acid signaling is required to maintain the homeostasis of mammary gland morphogenesis. PMID- 16217743 TI - Depression and suicide attempt as risk factors for incident unprovoked seizures. AB - Major depression has been shown to increase the risk for development of epilepsy, but prior studies have not evaluated whether this is due to specific symptoms of depression. We conducted a population-based case-control study of all newly diagnosed unprovoked seizures among Icelandic children and adults aged 10 years and older to test the hypothesis that major depression is a risk factor for developing unprovoked seizure and epilepsy, and to address whether specific symptoms of depression account for this increased risk. Cases were matched to the next two same sex births from the population registry. Using standardized interviews, we ascertained symptoms of major depression to make a Diagnostic and Statistical Manual, Fourth Edition (DSM-IV) diagnosis. A history of major depression was 1.7-fold more common among cases than among controls (95% confidence interval, 1.1-2.7). A history of attempted suicide was 5.1-fold more common among cases than among controls (95% confidence interval, 2.2-11.5). Attempted suicide increased seizure risk even after adjusting for age, sex, cumulative alcohol intake, and major depression or number of symptoms of depression. Major depression and attempted suicide independently increase the risk for unprovoked seizure. These data suggest that depression and suicide attempt may be due to different underlying neurochemical pathways, each of which is important in the development of epilepsy. PMID- 16217744 TI - Management of twin-twin transfusion syndrome in pregnancies with iatrogenic detachment of membranes following therapeutic amniocentesis and the role of interim amniopatch. AB - OBJECTIVE: Detachment of membranes may occur after therapeutic amniocentesis for twin-twin transfusion syndrome (TTTS). Subsequent amniocenteses or endoscopic fetal therapy may be hindered or made altogether impossible by this complication. The purpose of this study was to describe our experience in the assessment and management of TTTS patients with iatrogenic detached membranes (IDM). METHODS: Patients with IDM referred for fetal surgery for TTTS were considered ineligible for standard surgery and were offered different alternatives, including expectant management, serial amniocentesis, or an attempt at surgery with or without prior amniopatch. Pregnancy outcomes were compared between surgical and non-surgical patients. RESULTS: Nine hundred and forty-four patients with a diagnosis of TTTS were referred between July 1997 and December 2004, of whom 322 (34.1%) had a prior therapeutic amniocentesis. Twenty-six of the 322 patients (8%) had IDM. Ten patients opted to be managed with subsequent amniocenteses, two of which had an amniopatch. One patient had voluntary interruption of pregnancy. Fifteen patients underwent surgery, 10 of whom underwent an amniopatch. Overall, resealing of membranes occurred in 8/12 (66%) patients treated with an amniopatch. Survival of at least one fetus was greater in patients treated surgically with or without an amniopatch (12/15, 80% vs. 4/11, 36%, P = 0.04). CONCLUSION: Membrane detachment is an important complication of therapeutic amniocentesis in the treatment of TTTS. Although successful treatment of IDM can be achieved with an interim amniopatch, this alternative is not without risks. Therapeutic amniocenteses should be discouraged in patients considering endoscopic fetal surgery for TTTS. PMID- 16217745 TI - High-resolution real-time spiral MRI for guiding vascular interventions in a rabbit model at 1.5 T. AB - PURPOSE: To study the feasibility of a combined high spatial and temporal resolution real-time spiral MRI sequence for guiding coronary-sized vascular interventions. MATERIALS AND METHODS: Eight New Zealand White rabbits (four normal and four with a surgically-created stenosis in the abdominal aorta) were studied. A real-time interactive spiral MRI sequence combining 1.1 x 1.1 mm(2) in plane resolution and 189-msec total image acquisition time was used to image all phases of an interventional procedure (i.e., guidewire placement, balloon angioplasty, and stenting) in the rabbit aorta using coronary-sized devices on a 1.5 T MRI system. RESULTS: Real-time spiral MRI identified all rabbit aortic stenoses and provided high-temporal-resolution visualization of guide-wires crossing the stenoses in all animals. Angioplasty balloon dilatation and deployment of coronary-sized copper stents in the rabbit aorta were also successfully imaged by real-time spiral MRI. CONCLUSION: Combining high spatial and temporal resolution with spiral MRI allows real-time MR-guided vascular intervention using coronary-sized devices in a rabbit model. This is a promising approach for guiding coronary interventions. PMID- 16217746 TI - Heparanase promotes growth, angiogenesis and survival of primary breast tumors. AB - Despite great strides toward diagnosis and therapy, breast cancer remains a most threatening disease in its incidence, morbidity and mortality; therefore, additional knowledge regarding the molecular mechanisms contributing to breast cancer progression, as well as new targets for drug discovery are highly needed. Heparanase is the predominant enzyme involved in cleavage of heparan sulfate, the main polysaccharide component of the extracellular matrix. Experimental and clinical data indicate that heparanase plays important roles in cancer metastasis and angiogenesis. In breast carcinoma patients, heparanase expression correlates with the metastatic potential of the tumor. The present study was undertaken to investigate the role of heparanase in local growth and angiogenesis of primary breast tumors. MCF-7 breast carcinoma cells were stable transfected with the human heparanase (H-hpa) cDNA, or empty vector (mock), and injected into the mammary pad of nude mice. MRI was applied to monitor progression of tumor growth and angiogenesis. We demonstrate that tumors produced by cells overexpressing heparanase grew faster and were 7-fold larger than tumors produced by mock transfected cells. This enhanced growth was accompanied by increased tumor vascularization and a higher degree of vessel maturation. Histological examination ascribed the differences in tumor growth to heparanase-stimulated cell proliferation and survival. In-vitro experiments reinforced heparanase role as a survival factor under stress conditions. Moreover, H-hpa tumor cells infiltrate into the adjacent stroma, promoting formation of highly vascularized fibrous bands. Our results emphasize the significance and clarify the involvement of heparanase in primary breast cancer progression by generating a supportive microenvironment that promotes tumor growth, angiogenesis and survival. PMID- 16217747 TI - Gene expression profiling of 30 cancer cell lines predicts resistance towards 11 anticancer drugs at clinically achieved concentrations. AB - Cancer patients with tumors of similar grading, staging and histogenesis can have markedly different treatment responses to different chemotherapy agents. So far, individual markers have failed to correctly predict resistance against anticancer agents. We tested 30 cancer cell lines for sensitivity to 5-fluorouracil, cisplatin, cyclophosphamide, doxorubicin, etoposide, methotrexate, mitomycin C, mitoxantrone, paclitaxel, topotecan and vinblastine at drug concentrations that can be systemically achieved in patients. The resistance index was determined to designate the cell lines as sensitive or resistant, and then, the subset of resistant vs. sensitive cell lines for each drug was compared. Gene expression signatures for all cell lines were obtained by interrogating Affymetrix U133A arrays. Prediction Analysis of Microarrays was applied for feature selection. An individual prediction profile for the resistance against each chemotherapy agent was constructed, containing 42-297 genes. The overall accuracy of the predictions in a leave-one-out cross validation was 86%. A list of the top 67 multidrug resistance candidate genes that were associated with the resistance against at least 4 anticancer agents was identified. Moreover, the differential expressions of 46 selected genes were also measured by quantitative RT-PCR using a TaqMan micro fluidic card system. As a single gene can be correlated with resistance against several agents, associations with resistance were detected all together for 76 genes and resistance phenotypes, respectively. This study focuses on the resistance at the in vivo concentrations, making future clinical cancer response prediction feasible. The TaqMan-validated gene expression patterns provide new gene candidates for multidrug resistance. PMID- 16217748 TI - Do feathered dinosaurs exist? Testing the hypothesis on neontological and paleontological evidence. AB - The origin of birds and avian flight from within the archosaurian radiation has been among the most contentious issues in paleobiology. Although there is general agreement that birds are related to theropod dinosaurs at some level, debate centers on whether birds are derived directly from highly derived theropods, the current dogma, or from an earlier common ancestor lacking suites of derived anatomical characters. Recent discoveries from the Early Cretaceous of China have highlighted the debate, with claims of the discovery of all stages of feather evolution and ancestral birds (theropod dinosaurs), although the deposits are at least 25 million years younger than those containing the earliest known bird Archaeopteryx. In the first part of the study we examine the fossil evidence relating to alleged feather progenitors, commonly referred to as protofeathers, in these putative ancestors of birds. Our findings show no evidence for the existence of protofeathers and consequently no evidence in support of the follicular theory of the morphogenesis of the feather. Rather, based on histological studies of the integument of modern reptiles, which show complex patterns of the collagen fibers of the dermis, we conclude that "protofeathers" are probably the remains of collagenous fiber "meshworks" that reinforced the dinosaur integument. These "meshworks" of the skin frequently formed aberrant patterns resembling feathers as a consequence of decomposition. Our findings also draw support from new paleontological evidence. We describe integumental structures, very similar to "protofeathers," preserved within the rib area of a Psittacosaurus specimen from Nanjing, China, an ornithopod dinosaur unconnected with the ancestry of birds. These integumental structures show a strong resemblance to the collagenous fiber systems in the dermis of many animals. We also report the presence of scales in the forearm of the theropod ornithomimid (bird mimic) dinosaur, Pelecanimimus, from Spain. In the second part of the study we examine evidence relating to the most critical character thought to link birds to derived theropods, a tridactyl hand composed of digits 1-2-3. We maintain the evidence supports interpretation of bird wing digit identity as 2,3,4, which appears different from that in theropod dinosaurs. The phylogenetic significance of Chinese microraptors is also discussed, with respect to bird origins and flight origins. We suggest that a possible solution to the disparate data is that Aves plus bird-like maniraptoran theropods (e.g., microraptors and others) may be a separate clade, distinctive from the main lineage of Theropoda, a remnant of the early avian radiation, exhibiting all stages of flight and flightlessness. PMID- 16217749 TI - Arsenic trioxide synergizes with B7H3-mediated immunotherapy to eradicate hepatocellular carcinomas. AB - Arsenic trioxide (As(2)O(3)), a valuable anticancer drug for the treatment of acute promyelocytic leukemia, may also have therapeutic potential for the treatment of solid tumors. However, its therapeutic efficacy against solid tumors is lacking even at high dosages. Other therapeutic strategies are required to enhance the efficacy of As(2)O(3) against solid tumors such as hepatocellular carcinoma (HCC), which is refractory to chemotherapy. B7H3, a new member of the B7 family, has been shown to induce antitumor immunity. Intratumoral injection of B7H3 plasmids eradicates small EL-4 lymphomas, but monotherapy is ineffective against large tumors. Here we investigated whether As(2)O(3) would synergize with B7H3 immunotherapy to combat HCC. Large subcutaneous H22 HCCs (0.7-0.8 cm in diameter) established in BALB/c mice were rapidly and completely eradicated when intratumoral administration of As(2)O(3) was preceded by in situ gene transfer of B7H3. In contrast, neither As(2)O(3) nor B7H3 monotherapy was effective. The antitumor activity of As(2)O(3) was attributed to increased tumor-cell apoptosis, perhaps as a result of direct cytotoxicity as well as decreased tumor angiogenesis. Combination therapy generated potent systemic antitumor immunity mediated by CD8(+) and NK cells that was effective in combating a systemic challenge of 1 x 10(7) parental H22 cells. It led to the simultaneous and complete regression of multiple distant tumor nodules, concomitant with increased levels of serum IFN-gamma and cytotoxic T lymphocyte (CTL) activity. In conclusion, combining B7H3-mediated immunotherapy with As(2)O(3) warrants investigation as a therapeutic strategy to combat HCC, and other malignancies. PMID- 16217750 TI - siRNA gelsolin knockdown induces epithelial-mesenchymal transition with a cadherin switch in human mammary epithelial cells. AB - Epithelial-mesenchymal transition (EMT) describes a process occurring during development and oncogenesis by which epithelial cells obtain fibroblast-like properties and show reduced cell adhesion and increased motility. In this report, we demonstrated typical EMT in human mammary epithelial MCF10A small interfering (si)RNA gelsolin-knockdown cells. EMT was characterized by fibroblastic morphology, loss of contact inhibition and focus formation in monolayer growth, enhanced motility and invasiveness in vitro, increased actin filaments, overexpression of RAC, activation of both extracellular signal-regulated kinase and AKT, inactivation of glycogen synthase kinase-3, conversion of cadherin from the E- to N-type and induction of the transcription factor Snail. These results suggested that gelsolin functions as a switch that controls E- and N-cadherin conversion via Snail, and demonstrated that its knockdown leads to EMT in human mammary epithelial cells and possibly to the development of human mammary tumors. PMID- 16217751 TI - Endothelin system in oral squamous carcinoma cells: specific siRNA targeting of ECE-1 blocks cell proliferation. AB - The present study focused on the endothelin axis in human oral squamous cell carcinoma (SCC) cells. We investigated the expression and distribution of endothelin-1 (ET-1), its receptors (endothelin-A receptor (ET(A)R) and endothelin B receptor (ET(B)R)) and isoforms of its specific converting enzyme (ECE-1a, 1b, 1c) and the report on their relative influences on cell proliferation. We also investigated the effect of an ECE-specific inhibitor (ECE-i) and siRNA targeting of the ECE-1 gene on SCC cell proliferation. We observed the expression of ET-1, ET(A)R, ET(B)R and all endothelin-converting enzyme-1 (ECE-1) isoforms in oral SCC cells, but only the expression of ET-1, ET(B)R and ECE-1 was increased when compared to normal human epidermal keratinocytes. ET-1 alone stimulated proliferation of oral SCC cells. Antagonists of either ET(A)R or ET(B)R inhibited ET-1-mediated proliferation. Decreased ECE-1 expression after ECE siRNA treatment reduced SCC cell proliferation. Antiproliferative effects were also observed by inhibiting ECE activity with ECE-i. In conclusion, the present study demonstrates that modulation of the endothelin system in oral SCC cells might provide a novel therapeutic protocol for oral cancer. PMID- 16217752 TI - Visualization of episomal and integrated Epstein-Barr virus DNA by fiber fluorescence in situ hybridization. AB - For many Epstein-Barr virus (EBV)-associated malignancies, it is still a matter of controversy whether infected cells harbor episomal or chromosomally integrated EBV genomes or both. It is well established that the expression of EBV genes per se carries oncogenic potential, but the discrimination between episomal and integrated forms is of great relevance because integration events can contribute to the oncogenic properties of EBV, whereas host cells that exclusively harbor viral episomes may not carry the risks mediated by chromosomal integration. This notion prompted us to establish a reliable technique that not only allows to unequivocally discriminate episomal from integrated EBV DNA, but also provides detailed insights into the genomic organization of the virus. Here, we show that dynamic molecular combing of host cell DNA combined with fluorescence in situ hybridization (FISH) using EBV-specific DNA probes facilitate unambiguous discrimination of episomal from integrated viral DNA. Furthermore, the detection of highly elongated internal repeat 1 (IR1) sequences provides evidence that this method permits detection of major genomic alterations within the EBV genome. Thus, fiber FISH may also provide valuable insights into the genomic organization of viral genomes other than EBV. PMID- 16217753 TI - Targeting the epidermal growth factor receptor by erlotinib (Tarceva) for the treatment of esophageal cancer. AB - Esophageal cancer is the sixth most common cause of cancer-related death worldwide. Because of very poor 5-year survival new therapeutic approaches are mandatory. Erlotinib (Tarceva), an inhibitor of epidermal growth factor receptor tyrosine kinase (EGFR-TK), potently suppresses the growth of various tumors but its effect on esophageal carcinoma, known to express EGFR, remains unexplored. We therefore studied the antineoplastic potency of erlotinib in human esophageal cancer cells. Erlotinib induced growth inhibition of the human esophageal squamous cell carcinoma (ESCC) cell lines Kyse-30, Kyse-70 and Kyse-140, and the esophageal adenocarcinoma cell line OE-33, as well as of primary cell cultures of human esophageal cancers. Combining erlotinib with the EGFR-receptor antibody cetuximab, the insulin-like growth factor receptor tyrosine kinase inhibitor tyrphostin AG1024, or the 3-hydroxy-3-methylglutaryl coenzyme. A reductase (HMG CoAR) inhibitor fluvastatin resulted in additive or even synergistic antiproliferative effects. Erlotinib induced cell cycle arrest at the G1/S checkpoint. The erlotinib-mediated signaling involved the inactivation of EGFR-TK and ERK1/2, the upregulation of the cyclin-dependent kinase inhibitors p21(Waf1/CIP1) and p27(Kip1), and the downregulation of the cell cycle promoter cyclin D1. However, erlotinib did not induce immediate cytotoxicity or apoptosis in esophageal cancer cells. The inhibition of EGFR-TK by erlotinib appears to be a promising novel approach for innovative treatment strategies of esophageal cancer, as it powerfully induced growth inhibition and cell cycle arrest in human esophageal cancer cells and enhanced the antineoplastic effects of other targeted agents. PMID- 16217755 TI - Dietary carbohydrate, fibre, glycaemic index, glycaemic load and the risk of postmenopausal breast cancer. AB - Evidence that the insulin pathway may be involved in breast carcinogenesis has increased the interest in dietary factors that influence insulin secretion and resistance. We investigated dietary carbohydrate, fibre, glycaemic index (GI) and glycaemic load (GL) in a prospective study of 324 breast cancers diagnosed in 12,273 post-menopausal women. Although an increase of 1 standard deviation in carbohydrate was marginally associated with risk of breast cancer, relative risk (RR) 1.31 (95% CI, 0.98, 1.75), there were no significant associations with fibre, 1.08 (0.92, 1.26), GI, 0.98 (0.88, 1.10) or GL, 1.19 (0.93, 1.52) or with carbohydrate foods (bread, rice, pasta). The RR for carbohydrate and localized disease was elevated, 1.40 (1.02, 1.92), but like those for fibre, GI and GL did not differ significantly between localized and non-localized disease. RRs for grade I, but not grade II or III, tumours were elevated for fibre, 1.38 (1.08, 1.75), carbohydrate, 1.56 (1.08, 2.25) and GL, 1.41 (1.01, 1.98) but not for GI, 0.84 (0.65, 1.09). The RRs for fibre and oestrogen receptor (ER) positive (+) and progesterone receptor (PR) positive (+) tumours, 1.36 (1.10, 1.67), differed significantly from those for ER positive (+) and PR negative (-) tumours, 1.01 (0.61, 1.69) and ER-/PR- tumours, 0.65 (0.43, 0.99), p = 0.005. Our data do not support a strong role for GI and GL in breast carcinogenesis but suggest that increased intake of fibre and carbohydrate may be associated with the diagnosis of cancers of more favourable prognosis. PMID- 16217754 TI - The pro-cell death Bcl-2 family member, BNIP3, is localized to the nucleus of human glial cells: Implications for glioblastoma multiforme tumor cell survival under hypoxia. AB - The Bcl-2 nineteen kilodalton interacting protein 3 (BNIP3) is a hypoxia inducible proapoptotic member of the Bcl-2 family that induces cell death by associating with the mitochondria. Under normal conditions, BNIP3 is expressed in skeletal muscle and in the brain at low levels. In many human solid tumors, BNIP3 is upregulated in hypoxic regions but paradoxically, this BNIP3 expression fails to induce cell death. Herein, we have determined that BNIP3 is primarily localized to the nucleus of glial cells of the normal human brain, as well as in the malignant glioma cell line U251. Upon exposure of U251 cells to hypoxia, BNIP3 expression in the cytoplasm increases and localizes with the mitochondria, contributing to induction of cell death. In contrast, when BNIP3 is forcibly over expressed in the nucleus, it fails to induce cell death. Expression of N-terminal BNIP3 (lacking the transmembrane and conserved domains) in U251 cells blocks hypoxia-induced cell death acting as a dominant negative protein by binding to wild-type BNIP3 and blocking its association with the mitochondria. In glioblastoma multiforme (GBM) tumors, BNIP3 expression is increased in hypoxic regions of the tumor and is primarily localized to the nucleus in approximately 80% of tumors. Hence, BNIP3 is sequestered in the nucleus within the brain but under hypoxic conditions, BNIP3 becomes primarily cytoplasmic, promoting cell death. In GBMs, BNIP3 expression is increased but it remains sequestered in the nucleus in hypoxic regions, thereby blocking BNIP3's ability to associate with the mitochondria, providing tumor cells with a possible survival advantage. PMID- 16217756 TI - Selenium supplementation and colorectal adenomas: an analysis of the nutritional prevention of cancer trial. AB - Selenium status has been inversely associated with colorectal cancers (CRC) and adenomas. This investigation evaluates the association between selenium supplementation and prevalent and incident colorectal adenomas and CRC detected during the Nutritional Prevention of Cancer trial follow-up. Of the 1,312 randomized to 200 mcg of selenized yeast of matching placebo, 598 participants underwent endoscopic screening (flexible sigmoidoscopy or colonoscopy) for CRC sometime during the follow-up period, which ended in February 1, 1996. There was no colorectal screening performed at baseline. Of those screened, 77% were male (with a mean age of 62.8 years), 42% were former and 25% were current smokers. Adenomas were classified as prevalent (identified at the first endoscopic examination post-randomization during the follow-up period) or incident (identified at the second or subsequent examination). Ninety-nine prevalent and 61 incident adenomas were ascertained. Logistic regression odds ratios (OR) and 95% confidence intervals (CI) were calculated, adjusting for age, gender and smoking status. For prevalent adenomas, there was a suggestive but nonsignificant decrease in risk associated with selenium treatment (OR = 0.67, 95% CI = 0.43 1.05). Subjects in the lowest tertile of baseline selenium (OR = 0.27, 95% CI = 0.09-0.77) and current smokers (OR = 0.27, 95% CI = 0.11-0.66) had significant reductions in risk. The OR for incident adenomas was 0.98 (95% CI = 0.57-1.68). In addition to being associated with a reduced risk of incident CRC, selenium supplementation was associated with a significantly reduced risk of prevalent adenomas, but only among subjects with either a low baseline selenium level or among current smokers. PMID- 16217757 TI - Green tea catechin, epigallocatechin-3-gallate, inhibits vascular endothelial growth factor angiogenic signaling by disrupting the formation of a receptor complex. AB - A potential mechanism by which green tea may prevent cancer development is through the inhibition of angiogenesis. We have shown previously that the green tea catechin, epigallocatechin gallate (EGCG), inhibits endothelial cell tube formation through the inhibition of vascular endothelial growth factor (VEGF) induced Akt activation and vascular endothelial (VE)-cadherin phosphorylation. Furthermore, EGCG can suppress oxidant-induced production of the proangiogenic cytokine interleukin (IL)-8. To further elucidate the antiangiogenic mechanisms of EGCG, we investigated its regulation of other molecular processes in VEGF induced signaling in human umbilical vein endothelial cells (HUVECs). We show that EGCG at physiological doses (0.5-10 microM) markedly inhibits the formation of a vascular endothelial growth factor receptor 2 complex formed upon the binding of its ligand VEGF. This disruption results in a significant and dose dependent decrease in PI3-kinase activity. Electrophoretic mobility shift assay revealed that EGCG decreased the PI3 kinase-dependent activation and DNA-binding ability of NF-kappaB, likely acting through decreasing phosphorylation and degradation of IkappaB. VEGF-induced IL-8 production at the mRNA (real time RT PCR) and protein levels (ELISA) are also suppressed with EGCG. These results suggest a novel mechanism for green tea's anticancer effects where EGCG can abrogate VEGF signaling by interfering with the formation of a receptor complex, resulting in attenuated mitogenic and angiogenic signaling. PMID- 16217758 TI - Association of a distinctive strain of Epstein-Barr virus with gastric cancer. AB - Epstein-Barr virus (EBV) has been linked to gastric carcinoma (GC) with worldwide geographical variations attributable to types and variants of EBV. Here, we compare EBV strains between EBVaGC and healthy donors in Latin America, a high frequency area for EBVaGC. Tumor samples from 73 EBVaGC cases and throat washings from 329 healthy adults were examined for types 1 and 2 EBV and polymorphism at BamHI-F and BamHI-W1/I1 boundary regions and XhoI restriction site in LMP1 gene. Type 1 and prototype F of BamHI- F polymorphism accounted 59 (81%) and 69 (95%) of EBVaGC cases and 257 (78%) and 267 (81%) of healthy donors, respectively. Types I and "i" of BamHI W1/I1 polymorphism accounted 2 (3%) and 62 (85%) of EBVaGC and 85 (26%) and 170 (52%) of healthy donors, respectively (p<0.001). XhoI+ and - polymorphism accounted 60 (82%) and 4 (5%) of EBVaGC and 142 (43%) and 92 (28%) of healthy donors, respectively (p<0.001). Cosegregation analysis demonstrated that most of the 62 type "i" EBVaGC cases harbor XhoI+ strain (81%). However, among 143 type "i" healthy adults, both XhoI polymorphism were present in relatively similar frequencies (XhoI+ 58% and XhoI- 42%) (OR 9.0; 95% CI 1.2 69). Our findings are against to the proposed hypothesis that EBV strains are geographically but not disease-restricted. We conclude that most of the EBVaGC cases harbor a distinctive EBV strain (type "i"/XhoI +), but in healthy donors, this strain was as common as other strains. This finding is contrary to the proposed hypothesis that EBV strains are geographically but not disease restricted and identified a healthy population group that share the same strain that predominate in EBVaGC cases. PMID- 16217759 TI - Evaluation of cervical screening strategies with adjunct high-risk human papillomavirus testing for women with borderline or mild dyskaryosis. AB - The management of women with a smear read as borderline/mild dyskaryosis (BMD) found by cervical cancer screening is still under discussion as only few of these cases are associated with high-grade lesions. To determine the optimal screening strategy for these women, a simulation model of cervical cancer development was used that is based on high-risk human papillomavirus (hrHPV) infection. The current strategy of repeat cytological testing at 6 and 18 months after BMD was compared to strategies with adjunct hrHPV testing. Calculations were done for both conventional and liquid-based cytology as the primary screening tool. In comparison to current screening, adjunct hrHPV testing was more effective in preventing cancer and more woman-friendly (reduction in colposcopy referrals with outcome < cervical intraepithelial neoplasia (CIN2) of up to 56% and in repeat smears of 30-100%). In combination with conventional cytology, cost-effective strategies were the ones in which a sample for high-risk human papillomavirus (hrHPV) testing is collected at a return visit within 1 month or in which hrHPV testing is restricted to repeat smears taken at 6 and 18 months. For these strategies, co-collection of samples for hrHPV testing at baseline is not necessary which has organizational and cost advantages. In combination with liquid-based cytology, it was cost-effective to perform a reflex hrHPV test at baseline from the liquid-based specimen. Liquid-based screening was more effective than conventional screening, but annual diagnosis costs were euro5 million higher (population size 16 million). In conclusion, our calculations indicate that implementation of hrHPV testing for the management of women with borderline or mild dyskaryosis (BMD) is feasible both in settings where conventional and liquid-based cytology is current practice. PMID- 16217760 TI - Selective occlusion of tumor blood vessels by targeted delivery of an antibody photosensitizer conjugate. AB - The irregular vasculature and high interstitial pressure of solid tumors hinder the delivery of cytotoxic agents to cancer cells. As a consequence, the doses of chemotherapy necessary to achieve complete tumor eradication are associated with unacceptably high toxicities. The selective thrombosis of tumor blood vessels has been postulated as an alternative avenue for combating cancer, depriving tumors of nutrients and oxygen and causing an avalanche of tumor cell deaths. The human antibody L19, specific to the EDB domain of fibronectin, a marker of angiogenesis, is capable of selective in vivo localization around tumor blood vessels and is thus a suitable agent for delivering toxic payloads to the tumor neovasculature. Here we show that a chemical conjugate of the L19 antibody with the photosensitizer bis(triethanolamine)Sn(IV) chlorin e(6), after intravenous injection and irradiation with red light, caused an arrest of tumor growth in mice with subcutaneous tumors. By contrast, a photosensitizer conjugate obtained with an antibody of identical pharmacokinetic properties but irrelevant specificity did not exhibit a significant therapeutic effect. These results confirm that vascular targeting strategies, aimed at the selective occlusion/disruption of tumor blood vessels, have a significant anticancer therapeutic potential and encourage the use of antibody-photosensitizer conjugates for the therapy of superficial tumors and possibly other angiogenesis related pathologies. PMID- 16217761 TI - Immune stimulatory effects of CD70 override CD70-mediated immune cell apoptosis in rodent glioma models and confer long-lasting antiglioma immunity in vivo. AB - CD70 (CD27 ligand) promotes the expansion of primed lymphocytes by enhancing cell survival. Surprisingly, we previously observed that CD70 aberrantly expressed on human glioma cells promoted immune cell apoptosis and inhibited alloreactive lysis. Here we report that ectopic expression of CD70 in mouse glioma cells enhances apoptosis of T, B and NK cells in coculture, but nevertheless promotes glioma cell lysis by NK cells in vitro. In nude mice, CD70 expression in SMA-560 gliomas delays the glioma growth upon subcutaneous (s.c.) or intracerebral (i.c.) inoculation, suggesting a role for CD70/CD27-dependent NK cell activity in tumor surveillance. In syngeneic immunocompetent VM/Dk mice, CD70 allows the rejection of s.c. and i.c. implanted SMA-560 tumors. The tumorigenicity of CD70-expressing glioma cells is abrogated when TGF-beta signaling is blocked. Moreover, mice surviving the s.c. CD70 glioma challenge subsequently also reject wild-type glioma cells administered i.c. Similarly, CD70-expressing GL-261 gliomas are rejected in syngeneic C57BL/6 mice, while glioma growth is restored in C57BL/6 CD27(-/-) mice, suggesting that the CD70/CD27 interaction recruits a tumor specific T-cell repertoire and induces tumor-specific memory. Altogether, these observations indicate that the net effect of aberrant CD70 expression in gliomas is immune stimulatory rather than immune paralytic and encourage its application in tumor immunotherapy. PMID- 16217762 TI - Coffee and breast cancer risk. PMID- 16217763 TI - Ala228 variant of trail receptor 1 affecting the ligand binding site is associated with chronic lymphocytic leukemia, mantle cell lymphoma, prostate cancer, head and neck squamous cell carcinoma and bladder cancer. AB - Allelic loss of chromosome 8p21-22 is a frequent event in various human cancers including mantle cell lymphoma (MCL), prostate cancer, head and neck squamous cell carcinoma (HNSCC) and bladder cancer. The tumor necrosis factor-related apoptosis inducing ligand (TRAIL) receptors, including TNFRSF10A and TNFRSF10B, are located within this chromosomal region. Since recent studies demonstrate that chronic lymphocytic leukemia (CLL) and prostate cells are TRAIL induced apoptosis, TRAIL-receptors are strong tumor suppressor candidate genes in human cancers exhibiting loss of chromosomal material in 8p21.3. However, no mutation of the TRAIL receptor genes has been reported in CLL, MCL, prostate cancer, HNSCC so far. In this study we analyzed the complete coding region of TNFRSF10A and TNFRSF10B in a series of 32 MCL and 101 CLL samples and detected a single nucleotide polymorphism (SNP) in TNFRSF10A (A683C) with tumor specific allele distribution. We examined allele distribution in 395 samples of different tumor entities (prostate cancer, n = 43; HNSCC, n = 40; bladder cancer, n = 179) and compared them to 137 samples from healthy probands. We found the rare allele of TNFRSF10A is more frequent in CLL, MCL, prostate cancer, bladder cancer and HNSCC. The A683C polymorphism did not cosegregate with other TNFRSF10A polymorphisms previously described. Thus screening for 683A-->C nucleotide exchanges may become important in diagnosis and/or treatment of these malignancies. PMID- 16217764 TI - From gene profiling to diagnostic markers: IL-18 and FGF-2 complement CA125 as serum-based markers in epithelial ovarian cancer. AB - We used an oligonucleotide-based DNA microarray to identify potential markers in 39 primary cultures of ovarian cancer specimens compared with 11 primary cultures of normal ovarian epithelia. Differential gene expression of IL-18 and FGF-2 was validated on a subset of samples by quantitative PCR and by IHC, using an independent tissue array of 90 cores of 20 normal ovarian surface epithelia and 70 EOCs representing different grades and pathologies of ovarian disease. We further compared, by ELISA, these two markers with CA125 in sera from 25 cancer free and 47 ovarian cancer patients. IL-18 and FGF-2 proteins were significantly elevated in tumor tissues (p<0.04) and sera (p<0.05) from patients with ovarian cancer. In combination, the three markers (IL-18, FGF-2, and CA125) showed similar sensitivity in scoring for ovarian cancer (35/45 patients) compared to that of CA125 alone (37/45) and significantly improved the specificity of detection (20/25 patients) compared to each marker individually (15/25 for CA125; 18/25 FGF-2; 16/25 for IL-18). In conclusion we show that a combination of the three serum markers (IL-18, FGF-2 and CA125) is associated with EOC, with higher specificity than CA125 alone. Prospective studies with a large cohort of susceptible ovarian cancer patients will be required to expand these findings. PMID- 16217765 TI - Accelerated repair and reduced mutagenicity of oxidative DNA damage in human bladder cells expressing the E. coli FPG protein. AB - Repair of some oxidized purines such as 8-oxo-7,8-dihydroguanine (8-oxoG) is inefficient in human cells in comparison to repair of other major endogenous lesions (e.g. uracil, abasic sites or oxidized pyrimidines). This is due to the poor catalytic properties of hOGG1, the major DNA glycosylase involved in 8-oxoG removal. The formamidopyrimidine DNA glycosylase (FPG) protein from E. coli is endowed with a potent 8-oxoG glycolytic activity coupled with a beta,delta-AP lyase. In this study, we have expressed FPG fused to the enhanced green fluorescent protein (EGFP) in human bladder cells to accelerate the repair of oxidative DNA damage. Cells expressing the fusion protein EGFP-FPG repaired 8 oxoG and AP sites at accelerated rates, in particular via the single-nucleotide insertion base excision repair (BER) pathway and were resistant to mutagenicity of the oxidizing carcinogen potassium bromate. FPG may stably protect human cells from some harmful effects of oxidative DNA damage. PMID- 16217766 TI - Never smokers and lung cancer risk: a case-control study of epidemiological factors. AB - We performed an analysis of potential epidemiological risk factors for lung cancer using data from 280 cases and 242 hospital-based controls, all lifetime never smokers (those who had smoked <100 cigarettes in their lifetimes) and frequency matched on age, gender and ethnicity. The data on demographic characteristics, medical history of respiratory diseases (asthma, emphysema, pneumonia and hay fever), weight and height, family history, female characteristics and environmental tobacco smoke (ETS) and dust exposure were derived from personal interviews. We performed a logistic regression analysis of these variables adjusting for age, gender, ethnicity, income and years of education. Exposure to ETS (OR = 2.08, 95% CI [1.25-3.43]) and dusts (OR = 2.43, 95% CI [1.53-3.88]) were associated with significantly increased risk. In the analysis for joint effects, exposure to both ETS and dusts conferred a higher risk (OR = 3.25, 95% CI [1.58-6.70]) than exposure to either alone. Family history of any cancer with onset before age 50 in at least 1 first degree relative was a significant risk predictor (OR = 1.70, 95% CI [1.10-2.64]). Individuals with a self-reported physician-diagnosed history of hay fever, but not asthma, had a decreased lung cancer risk (OR = 0.57, 95% CI [0.35-0.92]). In the multivariate analysis, exposure to ETS and dusts, and family history of cancer with onset before age 50 were significant risk factors, while a history of hay fever (occurring without asthma) was significantly protective. PMID- 16217767 TI - Exploring SNP-SNP interactions and colon cancer risk using polymorphism interaction analysis. AB - Several single nucleotide polymorphisms (SNPs) in genes derived from distinct pathways are associated with colon cancer risk; however, few studies have examined SNP-SNP interactions concurrently. We explored the association between colon cancer and 94 SNPs, using a novel approach, polymorphism interaction analysis (PIA). We developed PIA to examine all possible SNP combinations, based on the 94 SNPs studied in 216 male colon cancer cases and 255 male controls, employing 2 separate functions that cross-validate and minimize false-positive results in the evaluation of SNP combinations to predict colon cancer risk. PIA identified previously described null polymorphisms in glutathione-S-transferase T1 (GSTT1) as the best predictor of colon cancer among the studied SNPs, and also identified novel polymorphisms in the inflammation and hormone metabolism pathways that singly or jointly predict cancer risk. PIA identified SNPs that may interact with the GSTT1 polymorphism, including coding polymorphisms in TP53 (Arg72Pro in p53) and CASP8 (Asp302His in caspase 8), which may modify the association between this polymorphism and colon cancer. This was confirmed by logistic regression, as the GSTT1 null polymorphism in combination with either the TP53 or the CASP8 polymorphism significantly alter colon cancer risk (p(interaction) < 0.02 for both). GSTT1 prevents DNA damage by detoxifying mutagenic compounds, while the p53 protein facilitates repair of DNA damage and induces apoptosis, and caspase 8 is activated in p53-mediated apoptosis. Our results suggest that PIA is a valid method for suggesting SNP-SNP interactions that may be validated in future studies, using more traditional statistical methods on different datasets. PMID- 16217768 TI - Inhibitory effects of unmethylated CpG oligodeoxynucleotides on MHC class I deficient and -proficient HPV16-associated tumours. AB - Unmethylated oligodeoxynucleotides containing guanine-cytidine dimers (CpG ODN) have been described as potent inducers of selected antitumour immune responses and the immunotherapeutic efficacy of CpG ODN has been examined either alone or as a vaccine adjuvant. We hypothesized that CpG ODN therapy could be an effective tool for immunotherapy of not only conventional MHC class I(+) tumours but also of those tumours that have lost MHC class I expression during their progression. To address this hypothesis, we employed the animal model resembling MHC class I proficient and -deficient human papilloma virus (HPV) 16-associated tumours. A cell line transformed with HPV16 E6 and E7 oncogenes, TC-1, as a prototype of MHC class I-positive line, and its MHC class I-deficient sublines TC-1/A9 and TC 1/P3C10 were injected into syngeneic C57BL/6 mice and the growing tumours were subjected to immunotherapy with CpG ODN 1826. The therapy started either 1 day after the challenge with the tumour cells or later, when the tumours had reached a palpable size. In both settings, CpG ODN 1826 significantly reduced the growth of MHC class I-proficient and -deficient tumours. Furthermore, we demonstrated that CpG ODN 1585, whose mechanism of action preferably involves indirect activation of the natural killer cells, induced regression of the MHC class I deficient tumours TC1/A9 but not of the MHC class I-proficient tumours TC-1. This study infers that synthetic CpG ODN have a potential for the therapy of both MHC class I-proficient and -deficient tumours and thus could be also used against tumours that tend to down-regulate their MHC class I expression. PMID- 16217769 TI - Bulky DNA adducts as risk indicator of lung cancer in a Danish case-cohort study. AB - Little is known of the predictive value of the levels of DNA adducts in terms of cancer risk. We examined the association between bulky DNA adducts and risk of lung cancer in a population-based cohort, comprising of 25,717 men and 27,972 women aged 50-64 years at entry. We included 245 cases (137 men and 108 women) with lung cancer and a comparison group of 255 individuals (137 men and 118 women), matched on sex, age and smoking duration. Bulky adducts in white blood cells collected at enrollment and stored at -150 degrees C were analyzed by (32)P postlabeling method, using the butanol enrichment procedure. The median level of bulky DNA adducts was 0.196 adduct/10(8) nucleotides (5-95 percentiles: 0.094 0.595) among current smokers who were later diagnosed with lung cancer and 0.163 adduct/10(8) nucleotides (5-95 percentiles: 0.091-0.455) among current smokers in the comparison group. The smoking adjusted incidence rate ratios (IRR) for lung cancer in relation to one log unit (natural logarithm) difference in adduct levels were 1.22 (95% CI 0.85-1.74), 1.33 (95% CI 0.89-1.98) and 0.76 (95% CI 0.39-1.47) among all, current and former smokers, respectively. Current smokers with bulky DNA adduct levels above the median had a significant higher lung cancer rate than those with adduct levels below the median (IRR = 1.61; 95% CI 1.04-2.49). The results are compatible with previous studies, suggesting a slightly higher risk of lung cancer with higher levels of adducts among smokers. Our results indicate that bulky DNA adducts may have a weak association with lung cancer risk. PMID- 16217770 TI - HER2 amplification in recurrent breast cancer following breast-conserving therapy correlates with distant metastasis and poor survival. AB - The authors analyzed the HER2 status in early-stage nonrecurrent and recurrent breast cancer groups following breast-conserving treatment. Retrospective analyses of a group of 36 invasive early breast cancer (IBC) patients who developed a local recurrence as a first event and of a random control group of 69 IBC patients were made. HER2 status was assessed by the HercepTest and fluorescence in situ hybridization. The Kaplan-Meier proportional log-rank test was used to study the impact of the biological factors on the metastasis-free interval (MFI) and the overall survival (OS). The Cox proportional hazards model, using stepwise selection was performed to identify the independent predictors of poor outcome. The median time of follow-up was 156 months (range: 22-230) for the nonrecurrent group of patients and 119 months (range: 36-228) for the recurrent group. No significant differences between either group were observed in terms of either patient or tumor characteristics, or of HER2 expression. However, a higher proportion of HER2 amplified cases were found in the recurrent group, in contrast to a higher proportion of hormonal receptor positive cases in the nonrecurrent group. After univariate and multivariate analyses, HER2 amplification was found to be an independent predictive factor for distant metastasis (HR = 10.75; p = 0.00008) and for survival (HR = 4.22; p = 0.004). In conclusion, HER2 amplification constitutes an independent poor prognostic factor for the MFI and OS in patients with recurrent breast cancer. The clinical implications are discussed. PMID- 16217771 TI - Identification of a novel human tissue factor splice variant that is upregulated in tumor cells. AB - Tissue factor (TF) is a transmembrane glycoprotein that serves as the prime initiator of blood coagulation and plays a critical role in thrombosis and hemostasis. In addition, a variety of tumor cells overexpress cell-surface TF, which appears to be important for tumor angiogenesis and metastasis. To elucidate the mechanism involved in the upregulation of TF in human tumor cells, a comprehensive analysis of TF mRNA from various normal and tumor cells was performed. The results of these studies indicate that, in addition to possessing a normal full-length TF transcript and minor levels of an alternatively spliced transcript known as alternatively-spliced tissue factor (asTF), human tumor cells express additional full-length TF transcripts that are also generated by alternative splicing. Reverse transcriptase-polymerase chain reaction (RT-PCR) and 5'-rapid amplification of cDNA ends- (5'-RACE) based analyses of cytoplasmic RNA from normal and tumor cells revealed that there is alternative splicing of the first intron between exon I and exon II resulting in 2 additional TF transcripts. One of the transcripts has an extended exon I with inclusion of most of the first TF intron (955 bp), while the second transcript is formed by the insertion of a 495 bp sequence, referred to as exon IA, derived from an internal sequence of the first intron. The full length TF transcript with alternatively spliced novel exon IA, referred to as alternative exon 1A-tissue factor (TF-A), represented approximately 1% of the total TF transcripts in normal cells, but constituted 7-10% of the total TF transcript in tumor cells. Quantitative real time RT-PCR analysis indicated that cultured human tumor cells contain 10-25-fold more copy numbers of TF-A in comparison to normal, untransformed cells. We propose that high-level expression of the novel TF-A transcript, preferentially in tumor cells, may have utility in the diagnosis and staging of a variety of solid tumors. PMID- 16217772 TI - Cigarette smoking and risk of glioma: a prospective cohort study. AB - The etiology of glioma, the most commonly diagnosed malignant brain tumor among adults in the United States, is poorly understood. N-nitroso compounds are known carcinogens, which are found in cigarette smoke and can induce gliomas in rats. On this basis, it has been hypothesized that cigarette smoking may be associated with an increased risk of glioma. We investigated the association between cigarette smoking and glioma risk in the National Breast Screening Study, which included 89,835 Canadian women aged 40-59 years at recruitment between 1980 and 1985. Linkages to national cancer and mortality databases yielded data on cancer incidence and deaths from all causes, respectively, with follow-up ending between 1998 and 2000. Cox proportional hazard models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for the association between cigarette smoking and risk of glioma. During a mean of 16.4 years of follow-up, we observed 120 incident glioma cases. Among ever smokers, women who reported having quit smoking had a 51% increase in risk of glioma compared with never smokers (HR = 1.51, 95% CI = 0.97-2.34), while current smokers did not appear to have an increase in risk. When the association with former smokers was further examined by years since quitting, women who had quit smoking >10 years before baseline were at a decreased risk of glioma compared with women who had quit within the 10 years prior to baseline (HR = 0.55, 95% CI = 0.29-1.07), indicating that the association between former smokers and glioma may be driven by women, who recently quit smoking. Compared with nonsmokers, duration of cigarette smoking, number of cigarettes smoked per day and pack-years of smoking were associated with increased glioma risk, although the increases in risk were relatively modest. The present study provides some support for a positive association between cigarette smoking and risk of glioma. PMID- 16217773 TI - Quantifying the effect of posture on intracranial physiology in humans by MRI flow studies. AB - PURPOSE: To quantify the effect of posture on intracranial physiology in humans by MRI, and demonstrate the relationship between intracranial compliance (ICC) and pressure (ICP), and the pulsatility of blood and CSF flows. MATERIALS AND METHODS: Ten healthy volunteers (29+/-7 years old) were scanned in the supine and sitting positions using a vertical gap MRI scanner. Pulsatile blood and CSF flows into and out from the brain were visualized and quantified using time-of-flight (TOF) and cine phase-contrast techniques, respectively. The total cerebral blood flow (tCBF), venous outflow, ICC, and ICP for the two postures were then calculated from the arterial, venous, and CSF volumetric flow rate waveforms using a previously described method. RESULTS: In the upright posture, venous outflow is considerably less pulsatile (57%) and occurs predominantly through the vertebral plexus, while in the supine posture venous outflow occurs predominantly through the internal jugular veins. A slightly lower tCBF (12%), a considerably smaller CSF volume oscillating between the cranium and the spinal canal (48%), and a much larger ICC (2.8-fold) with a corresponding decrease in the MRI-derived ICP values were measured in the sitting position. CONCLUSION: The effect of posture on intracranial physiology can be quantified by MRI because posture related changes in ICC and ICP strongly affect the dynamics of cerebral blood and CSF flows. This study provides important insight into the coupling that exists between arterial, venous, and CSF flow dynamics, and how it is affected by posture. PMID- 16217775 TI - Strategies for shimming the breast. AB - There is evidence in the literature indicating a significant static field inhomogeneity in the human breast. A nonhomogenous field results in line broadening and frequency shifts in MRS and can cause intensity loss and spatial errors in MRI. Thus, there is a clear rationale for determining the regional variations in the static field homogeneity in the breast and providing strategies to correct them. Herein, the nature and extent of the static magnetic field at 3 T were measured in central planes of the human breast using both phase maps and multivoxel MRS techniques. In addition, the effect of first- and high-order shimming and of spatial saturation pulses on the static field inhomogeneity was evaluated. Both the theoretical and the measured field were found to be primarily linear in nature, with a reduction of 300 Hz from the nipple to the chest wall. First-order shimming reduced this inhomogeneity by 65%. Interestingly, the combination of spatial saturation pulses and first-order shimming was more effective than high-order shim alone. Since many clinical scanners do not have either higher-order shim or automated higher shimming algorithms that work in the presence of fat, the suggested combination provides an effective means to correct inhomogeneities in the breast. PMID- 16217776 TI - Magnetic resonance imaging-based spirometry for regional assessment of pulmonary function. AB - In this work MRI-based spirometry is presented as a method for noninvasively assessing pulmonary mechanical function on a regional basis. A SPAMM tagging sequence was modified to allow continuous dynamic imaging of the lungs during respiration. A motion-tracking algorithm was developed to track material regions from time-resolved grid-tagged images. Experiments were performed to image the lungs during quiet breathing and volumetric strain was calculated from the measured displacement maps. Regional volume calculations, derived from volumetric strain, were integrated over the entire lung and compared to segmented volume calculations with good agreement. Results from this work demonstrate that MRI spirometry has the potential to become a clinically useful tool for measuring regional ventilation and assessing pulmonary diseases that regionally affect the mechanical function of the lung. PMID- 16217777 TI - Interleaved acquisition of lipid and water images of the heart using a double inversion fast spin-echo method. AB - In this work we present a new method for the improved detection of lipid infiltration in the heart. The method employs a double-inversion fast spin-echo technique where the acquisition of water- and lipid-suppressed k-space data is alternated between TR periods to produce co-registered lipid and water images from data acquired in a breath hold. The lipid and water images can then be combined to generate a lipid/water image with reduced artifacts due to flow and excellent contrast between lipid and myocardium. The method is demonstrated in ex vivo tissue and in vivo. This novel method may improve the detection of lipid infiltration in the heart in pathologies such as arrhythmogenic right ventricular dysplasia. PMID- 16217778 TI - Equivalent cross-relaxation rate imaging for sentinel lymph node biopsy in breast carcinoma. AB - Sentinel lymph node biopsy (SLNB) is an important technique for detecting axillary lymph node metastasis in breast carcinoma patients. However, false negative results are a problem. Equivalent cross-relaxation rate (ECR) imaging (ECRI) is a measurement method that can be used to quantitatively evaluate a change in the structural organization of lymph nodes by magnetic resonance imaging (MRI). We performed axillary ECRI in an attempt to decrease the false negative results of SLNB. Regions without metastases showed a higher ECR value. On the other hand, regions with metastases showed a lower ECR value. The ECR images were compared with macroscopic histology images in which the presence or absence of axillary lymph node metastasis could be evaluated. ECRI is a potentially useful method for evaluating the efficacy of SLNB. PMID- 16217779 TI - Current usage and attitudes among interventional cardiologists regarding the performance of percutaneous coronary intervention (PCI) in the outpatient setting. PMID- 16217780 TI - Nuclear magnetic resonance parameters for monitoring coagulation of liver tissue. AB - A new NMR parameter is suggested as a sensitive tool for monitoring thermal coagulation of liver tissue. That parameter is the proton magnetization exchange time (tau(MEX)) between water and the proteins. tau(MEX) was very sensitive to coagulation and insensitive to temperature, therefore representing only damage to the tissue, independent of effects caused by temperature fluctuations. The measurement of tau(MEX) by two different methods revealed the existence of two or more groups of proteins, characterized by their different transverse relaxation time, and tau(MEX). PMID- 16217781 TI - Novel rapid fat suppression strategy with spectrally selective pulses. AB - Short repetition time gradient echo sequences are gaining popularity in clinical applications such as dynamic contrast enhancement imaging, cardiac imaging, and MR angiography. Performing fat suppression in these sequences is usually time consuming and often somewhat ineffective, due to the relatively short T(1) and long T(2) of fat. A novel rapid fat suppression strategy using spectrally selective pulses is introduced and compared with clinically popular sequences such as fat presaturated fast field echo (FFE) and turbo field echo (TFE) and binomial water-selective spatial-spectral excitation (SSE, or SPSP excitation) FFE. The new strategy combines fat presaturation with low-order binomial water selective SSE pulses in a TFE sequence. This enables the use of a long echo train length to decrease exam time, but without creation of excess fat signal contamination of the resultant images. The fat nullification is also more reliable as fat signals in central k-space data are suppressed twice. An implementation of this strategy is compared with traditional methods in both phantom and human studies, confirming that the new technique provides strong fat suppression with few artifacts despite the short scan duration. PMID- 16217782 TI - In vivo sodium magnetic resonance imaging of the human brain using soft inversion recovery fluid attenuation. AB - Sodium imaging with soft inversion recovery fluid attenuation, which may be advantageous for intracellular weighting, was demonstrated with cerebrospinal fluid (CSF) suppression in five healthy volunteers at 4.7 T. Long rectangular inversion pulses reduce the average power deposition in an inversion recovery sequence, allowing repetition time to be shortened and more averages acquired for a given scan length. Longer pulses also significantly reduce the "depth" of Mz inversion in environments with rapid T1 and T2 relaxation (i.e., brain relative to CSF). Phantom experiments and simulation show a marked SNR increase when using a 10-ms, rather than a 1-ms, rectangular inversion pulse. Images were acquired in 11.1 min with a voxel size of 0.25 cm3 and the SNR in CSF, which is typically approximately 3 times larger than in brain, was reduced to 23% of that in the brain tissue, which had an average SNR of 17. PMID- 16217783 TI - T1rho contrast in functional magnetic resonance imaging. AB - The application of T1 in the rotating frame (T1rho) to functional MRI in humans was studied at 3 T. Increases in neural activity increased parenchymal T1rho. Modeling suggested that cerebral blood volume mediated this increase. A pulse sequence named spin-locked echo planar imaging (SLEPI) that produces both T1rho and T2* contrast was developed and used in a visual functional MRI (fMRI)experiment. Spin-locked contrast significantly augments the T2* blood oxygen level-dependent (BOLD) contrast in this sequence. The total functional contrast generated by the SLEPI sequence (1.31%) was 54% larger than the contrast (0.85%) obtained from a conventional gradient-echo EPI sequence using echo times of 30 ms. Analysis of image SNR revealed that the spin-locked preparation period of the sequence produced negligible signal loss from static dephasing effects. The SLEPI sequence appears to be an attractive alternative to conventional BOLD fMRI, particularly when long echo times are undesirable, such as when studying prefrontal cortex or ventral regions, where static susceptibility gradients often degrade T2*-weighted images. PMID- 16217784 TI - Comparison of X-ray fluoroscopy and interventional magnetic resonance imaging for the assessment of coronary artery stenoses in swine. AB - The accuracy of a two-step interventional MRI protocol to quantify coronary artery disease was compared to the clinical gold standard, X-ray angiography. Studies were conducted in nine swine with a surgically induced stenosis in the proximal left circumflex coronary artery. The two-step protocol consisted of catheter-directed magnetic resonance angiography (MRA), which was first used to localize the stenosis, followed by MRI cross-sectional images to quantify the degree of stenosis without the use of contrast agent. Line signal intensity profiles were drawn across the vessel diameter at the stenosis site and proximal to the stenosis for each data set to measure percentage stenosis for each animal. Catheter-directed MRA successfully detected eight of nine stenoses. Cross sectional MRI accurately quantified each stenosis, with strong agreement to the measurements made using X-ray fluoroscopy (intraclass correlation coefficient = 0.955; P < 0.05). This study demonstrates that in the future interventional MRI may be an alternative to X-ray angiography for the detection and quantification of coronary artery disease. PMID- 16217785 TI - Myocardial first pass perfusion: steady-state free precession versus spoiled gradient echo and segmented echo planar imaging. AB - The imaging sequences used in first pass (FP) perfusion to date have important limitations in contrast-to-noise ratio (CNR), temporal and spatial resolution, and myocardial coverage. As a result, controversy exists about optimal imaging strategies for FP myocardial perfusion. Since imaging performance varies from subject to subject, it is difficult to form conclusions without direct comparison of different sequences in the same subject. The purpose of this study was to directly compare the saturation recovery SSFP technique to other more commonly used myocardial first pass perfusion techniques, namely spoiled GRE and segmented EPI. Differences in signal-to-noise ratio (SNR), CNR, relative maximal upslope (RMU) of signal amplitude, and artifacts at comparable temporal and spatial resolution among the three sequences were investigated in computer simulation, contrast agent doped phantoms, and 16 volunteers. The results demonstrate that SSFP perfusion images exhibit an improvement of approximately 77% in SNR and 23% in CNR over spoiled GRE and 85% SNR and 50% CNR over segmented EPI. Mean RMU was similar between SSFP and spoiled GRE, but there was a 58% increase in RMU with SSFP versus segmented EPI. PMID- 16217786 TI - Fat/water separation in single acquisition steady-state free precession using multiple echo radial trajectories. AB - Phase detection in fully refocused SSFP imaging has recently allowed fat/water separation without preparing the magnetization or using multiple acquisitions. Instead, it exploits the phase difference between fat and water at an echo time at the midpoint of the TR. To minimize the TR for improved robustness to B0 inhomogeneity, a 3D projection acquisition collecting two half echoes at the beginning and end of each excitation was previously implemented. Since echoes are not formed at the midpoint of the TR, this method still requires two passes of k space for fat/water separation. A new method is presented to linearly combine the half echoes to separate fat and water in a single acquisition. Separation using phase detection provides superior contrast between fat and water voxels. Results from high resolution angiography and musculoskeletal studies with improved robustness to inhomogeneity and a 50% scan time reduction compared to the two pass method are presented. PMID- 16217787 TI - Diagnostic discordance of electromyography (EMG) versus voiding cystourethrogram (VCUG) for detrusor-external sphincter dyssynergy (DESD). AB - AIMS: Detrusor sphincter dyssynergia (DSD) is defined as: "a detrusor contraction concurrent with an involuntary contraction of the urethral and/or periurethral striated muscle." In neurogenic etiology, this usually refers to involuntary contraction of the external striated sphincter and has classically been termed detrusor-external sphincter dyssynergia (DESD). There is currently no consensus regarding diagnosis [specifics of electromyographic (EMG) or voiding cystourethrographic (VCUG) determination], and little data on how well these modalities correlate. We explore the diagnostic congruence for DESD between needle EMG and VCUG in the neurogenic population. METHODS: Consecutive studies performed by a single urodynamicist at a major neurologic center were reviewed. Presence of DESD was determined by increased wire needle EMG activity and/or by dilated bladder neck and proximal urethra during detrusor contraction, in the absence of valsalva or attempt to inhibit voiding. Minimal acceptable criterion for agreement between the two tests was set at 70%. RESULTS: Fourty nine patients were diagnosed with DESD, had a videourodynamic study available, and had no history of sphincterotomy or stent. Binomial testing demonstrated significant disagreement (P < 0.000) in observed proportions. There was 60% agreement (28 patients) and 40% disagreement (21 patients) between EMG and VCUG for diagnosis of DESD. CONCLUSIONS: We found significant disagreement between needle EMG and VCUG for a positive diagnosis of DESD. A combination of EMG and VCUG may identify more cases of DESD than either modality alone and underscores the need for more strict criteria when defining this entity from a urodynamic standpoint. PMID- 16217789 TI - Patterns of axonal branching of neurons of the substantia nigra pars reticulata and pars lateralis in the rat. AB - Axons from neurons of the rat substantia nigra pars reticulata (SNr) and pars lateralis (SNl) were traced after injecting their cell body with biotinylated dextran amine. Thirty-two single axons were reconstructed from serial sagittal sections with a camera lucida, whereas four other SNr axons were reconstructed in the coronal plane to determine whether they innervate the contralateral hemisphere. Four distinct types of SNr projection neurons were identified based on their main axonal targets: type I neurons that project to the thalamus; type II neurons that target the thalamus, the superior colliculus (SC), and the pedunculopontine tegmental nucleus (PPTg); type III neurons that project to the periaqueductal gray matter and the thalamus; and type IV neurons that target the deep mesencephalic nucleus (DpMe) and the SC. The axons of the SNl showed the same branching patterns as SNr axons of types I, II, and IV. The coronal reconstructions demonstrated that SNr neurons innervate the thalamus, the SC, and the DpMe bilaterally. At the thalamic level, SNr and SNl axons targeted preferentially the ventral medial, ventral lateral, paracentral, parafascicular, and mediodorsal nuclei. Axons reaching the SC arborized selectively within the deep layers of this structure. Our results reveal that the SNr and SNl harbor several subtypes of projection neurons endowed with a highly patterned set of axon collaterals. This organization allows single neurons of these output structures of the basal ganglia to exert a multifaceted influence on a wide variety of diencephalic and midbrain structures. PMID- 16217788 TI - Expression of the zebrafish Iroquois genes during early nervous system formation and patterning. AB - Iroquois genes are involved in many patterning processes during development. In particular, they act as prepattern genes to control proneural gene expression both in Drosophila and in vertebrates. In this paper, we have analyzed the expression during embryogenesis of the 11 zebrafish Iroquois genes, with special interest for nervous system formation and patterning. During the first 2 days of development, Iroquois genes are expressed in distinct domains in the neuroepithelium, as well as in groups of neuronal progenitors and neurons. They are also expressed at different stages of placodal development. These expression patterns are similar to the patterns of the murine irx genes and also show features specific to teleosts. For the zebrafish Iroquois gene family, we find both specific patterns and patterns conserved within a cluster, between paralogues, or in most genes of the family. Overall, these expression data suggest functions for the Iroquois family of transcription factors in neural and placodal patterning, neurogenesis, and neuronal specification. PMID- 16217790 TI - Coexpression of cholinergic and noradrenergic phenotypes in human and nonhuman autonomic nervous system. AB - It has long been known that the sympathetic innervation of the sweat glands is cholinergic in most mammalian species and that, during development, rodent sympathetic cholinergic sweat gland innervation transiently expresses noradrenergic traits. We show here that some noradrenergic traits persist in cholinergic sympathetic innervation of the sweat glands in rodents but that lack of expression of the vesicular monoamine transporter renders these cells functionally nonnoradrenergic. Adult human sweat gland innervation, however, is not only cholinergic but coexpresses all of the proteins required for full noradrenergic function as well, including tyrosine hydroxylase, aromatic amino acid decarboxylase, dopamine beta-hydroxylase, and the vesicular monoamine transporter VMAT2. Thus, cholinergic/noradrenergic cotransmission is apparently a unique feature of the primate autonomic sympathetic nervous system. Furthermore, sympathetic neurons innervating specifically the cutaneous arteriovenous anastomoses (Hoyer-Grosser organs) in humans also possess a full cholinergic/noradrenergic cophenotype. Cholinergic/noradrenergic coexpression is absent from other portions of the human sympathetic nervous system but is extended in the parasympathetic nervous system to intrinsic neurons innervating the heart. These observations suggest a mode of autonomic regulation, based on corelease of norepinephrine and acetylcholine at parasympathocardiac, sudomotor, and selected vasomotor neuroeffector junctions, that is unique to the primate peripheral nervous system. PMID- 16217792 TI - Bilateral symmetric organization of neural elements in the visual system of a coelenterate, Tripedalia cystophora (Cubozoa). AB - Cubozoans differ from other cnidarians by their body architecture and nervous system structure. In the medusa stage they possess the most advanced visual system within the phylum, located in sophisticated sensory structures, rhopalia. The rhopalium is a club-shaped structure with paired pit-shaped pigment cup eyes, paired slit-shaped pigment cup eyes, and two complex camera-type eyes: one small upper lens eye and one large lower lens eye. The medusa carries four rhopalia and visual processing and locomotor rhythm generation takes place in the rhopalia. We show here a bilaterally symmetric organization of neurons, with commissures connecting the two sides, in the rhopalium of the cubozoan Tripedalia cystophora. The fortuitous observation that a subset of neurons is strongly immunoreactive for a PCNA (proliferating cell nuclear antigen)-like epitope allowed us to analyze the organization of these neurons in detail. Distinct PCNA-immunoreactive (PCNA-ir) nuclei form six bilateral pairs that are associated with the slit eyes, pit eyes, upper lens eye, and the posterior wall of the rhopalium. Three commissures connect the clusters of the two sides and all clusters in the rhopalium have connections to the area around the base of the stalk. This neuronal system provides an anatomical substrate for integration of visual signals from the different eyes. PMID- 16217794 TI - Nicotinamide promotes long-term survival and extensive neurite outgrowth in ultimobranchial C cells cultured from chick embryos. AB - In avian species, the ultimobranchial anlage is populated with neuronal cells derived from the distal vagal ganglion. We found that ultimobranchial C cells of chick embryos cultured in the presence of nicotinamide continued to grow for at least 60 days and exhibited profound morphological changes, resulting in the formation of dense networks of neuronal fibers. Nicotinamide, thus, facilitated the manifestation of neuronal features in C cells. The neuronal phenotypes of cultured C cells were analyzed in detail by both scanning and transmission electron microscopy. Their neural nature was also positively established by immunostaining with monoclonal antibodies to the neuronal markers neuron-specific class III beta-tubulin (TuJ1), microtubule-associated protein (MAP) 2, and synaptophysin. Confocal laser scanning microscopy confirmed that these neuron specific proteins are colocalized with calcitonin in both the somata and the neuronal processes of C cells. Furthermore, reverse transcriptase-polymerase chain reaction analyses, performed at various times up to 30 days in culture, indicated that the C cells have persistent gene expression of calcitonin, the catecholamine-synthesizing enzyme tyrosine hydroxylase, proenkephalin, proopiomelanocortin, neuron-specific beta-tubulin (cbeta4), SCG10, and Bcl-2. The morphological responses of C cells to nicotinamide treatment were analyzed quantitatively over a period of 60 days. The area of C-cell colonies, number of processes per colony, and length of processes continued to increase until culture day 45. In conclusion, nicotinamide stimulates long-term survival and neuronal differentiation of chick embryo C cells, and this culture system may provide a useful model for studying neuronal differentiation mechanisms. PMID- 16217793 TI - Alpha-9 nicotinic acetylcholine receptor immunoreactivity in the rodent vestibular labyrinth. AB - Vestibular tissues (cristae ampullares, macular otolithic organs, and Scarpa's ganglia) in chinchilla, rat, and guinea pig were examined for immunoreactivity to the alpha9 nicotinic acetylcholine receptor (nAChR) subunit. The alpha9 antibody was generated against a conserved peptide present in the intracellular loop of the predicted protein sequence of the guinea pig alpha9 nAChR subunit. In the vestibular periphery, staining was observed in calyces around type I hair cells, at the synaptic pole of type II hair cells, and in varying levels in Scarpa's ganglion cells. Ganglion cells were also triply labeled to detect alpha9, calretinin, and peripherin. Calretinin labels calyx-only afferents. Peripherin labels bouton-only afferents. Dimorphic afferents, which have both calyx and bouton endings, are not labeled by calretinin or peripherin. In these experiments, alpha9 was expressed in both calyx and dimorphic afferents. A subpopulation of small ganglion cells did not contain the alpha9 nAChR but did stain for peripherin. We surmise that these are bouton-only afferents. Bouton (regularly discharging) afferents also show efferent responses, although they are qualitatively different from those in irregularly discharging (calyx and dimorphic) afferents, much slower and longer lasting. Thus, regular afferents are probably more affected via a muscarinic cholinergic or a peptidergic mechanism, with a much smaller superimposed fast nicotinic-type response. This latter response could be due to one of the other nicotinic receptors that have been described in studies from other laboratories. PMID- 16217796 TI - Two parts of the nucleus prepositus hypoglossi project to two different subdivisions of the dorsolateral periaqueductal gray in cat. AB - The dorsolateral column of the mesencephalic periaqueductal gray (PAG) is a separate part of the PAG. Its afferent sources, efferent targets, and neurochemical properties differ from the adjacent PAG columns. The dorsolateral PAG is thought to be associated with aversive behaviors, but it is not yet understood how these behaviors are brought about. To elucidate the function of the PAG further, in the present study we investigated which brainstem regions project to the dorsolateral PAG. Wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injections involving the dorsolateral PAG, but extending into the lateral part, resulted in many retrogradely labeled cells in the pontine and medullary tegmentum bilaterally. However, it was concluded that these neurons were labeled from the lateral PAG, because no anterograde labeling was found in the dorsolateral PAG after a large injection into the tegmentum. Retrogradely labeled cells were also found in the nucleus prepositus hypoglossi (PPH), mainly contralaterally. Injections of [3H]leucine or WGA-HRP in the PPH resulted in anterogradely labeled fibers in the dorsolateral PAG. Two separate distribution patterns were found. The caudal and intermediate PPH projected to a small region on the dorsolateral edge of the dorsolateral column, whereas the supragenual PPH distributed labeled fibers to all other parts of the dorsolateral PAG, except the area on the dorsolateral edge. These separate PPH projections suggest that two subdivisions exist within the dorsolateral PAG. The present findings suggest a role for the dorsolateral PAG in the oculomotor system. PMID- 16217795 TI - Postnatal changes of vesicular glutamate transporter (VGluT)1 and VGluT2 immunoreactivities and their colocalization in the mouse forebrain. AB - Vesicular glutamate transporter 1 (VGluT1) and VGluT2 accumulate neurotransmitter glutamate into synaptic vesicles at presynaptic terminals, and their antibodies are thus considered to be a good marker for glutamatergic axon terminals. In the present study, we investigated the postnatal development and maturation of glutamatergic neuronal systems by single- and double-immunolabelings for VGluT1 and VGluT2 in mouse forebrain including the telencephalon and diencephalon. VGluT2 immunoreactivity was widely distributed in the forebrain, particularly in the diencephalon, from postnatal day 0 (P0) to adulthood, suggesting relatively early maturation of VGluT2-loaded glutamatergic axons. In contrast, VGluT1 immunoreactivity was intense only in the limbic regions at P0, and drastically increased in the other telencephalic and diencephalic regions during three postnatal weeks. Interestingly, VGluT1 immunoreactivity was frequently colocalized with VGluT2 immunoreactivity at single axon terminal-like profiles in layer IV of the primary somatosensory area from P5 to P10 and in the ventral posteromedial thalamic nucleus from P0 to P14. This was in sharp contrast to the finding that almost no colocalization was found in glomeruli of the olfactory bulb, patchy regions of the caudate-putamen, and the ventral posterolateral thalamic nucleus, where moderate to intense immunoreactivities for VGluT1 and VGluT2 were intermingled with each other in neuropil during postnatal development. The present results indicate that VGluT2-loaded glutamatergic axons maturate earlier than VGluT1-laden axons in the mouse telencephalic and diencephalic regions, and suggest that VGluT1 plays a transient developmental role in some glutamatergic systems that mainly use VGluT2 in the adulthood. PMID- 16217798 TI - Dmrt1 expression in response to estrogen treatment in a reptile with temperature dependent sex determination. AB - Dmrt1 has been implicated as an important factor in sex determination in all classes of vertebrates, including reptiles with temperature-dependent sex determination (TSD). Specifically, early embryonic expression of Dmrt1 appears to be an integral part of normal testicular development in vertebrates. Recently, a number of TSD studies have placed Dmrt1 expression at the top of a short list of putative temperature-sensitive events for TSD. Dmrt1 expression has been shown to be up-regulated at male-producing temperatures during the thermosensitive period (TSP) of several TSD reptile species. An interesting finding in Dmrt1 studies of fish and amphibians has been that in species where exogenous steroids can stimulate sex reversal, Dmrt1 expression can also be manipulated by these steroid treatments. In the current study, we examine the effects of exogenous 17beta estradiol treatment on Dmrt1 expression at male-producing temperature (26 degrees C) in a reptile with TSD, the red-eared slider turtle, Trachemys scripta. Previous studies have demonstrated that exogenous estrogens can stimulate sex reversal (at male-producing temperatures) in this species of turtle. In the current study, T. scripta embryos that received estradiol treatment displayed a significant decrease in Dmrt1 expression during the TSP, while embryos in the control group (no estradiol treatment) showed significant increases in Dmrt1 expression during the TSP. Furthermore, the pattern of Dmrt1 expression for the estradiol-treated group was similar to the pattern we previously reported for Dmrt1 at female-producing temperature (31 degrees C). These findings indicate that exogenous estrogen is acting at or before the Dmrt1 event in the sex determination/sex differentiation cascade of T. scripta. Further, the results are consistent with the hypothesis that exogenous estrogen could be exerting its feminizing effect by down-regulating the expression of Dmrt1. PMID- 16217797 TI - Isolation of the feline alpha1,3-galactosyltransferase gene, expression in transfected human cells and its phylogenetic analysis. AB - The enzyme alpha 1,3-galactosyltransferase (alpha1,3-GT), which catalyzes synthesis of terminal alpha-galactosyl epitopes (Gal alpha1,3Gal beta1-4GlcNAc R), is produced in non-primate mammals, prosimians and new-world monkeys, but not in old-world monkeys, apes and humans. We cloned and sequenced a cDNA that contains the coding sequence of the feline alpha1,3-GT gene. Flow cytometric analysis demonstrated that the alpha-galactosyl epitope was expressed on the surface of a human cell line transduced with an expression vector containing this cDNA, and this alpha-galactosyl epitope expression subsided by alpha galactosidase treatment. The open reading frame of the feline alpha1,3-GT cDNA is 1,113 base pairs in length and encodes 371 amino acids. The nucleotide sequence and its deduced amino acid sequence of the feline alpha1,3-GT gene are 88-90% and 85-87%, respectively, similar to the reported sequences of the bovine, porcine, marmoset and cebus monkey alpha1,3-GT genes, while they are 88% and 82-83%, respectively, similar to those of the orangutan and human alpha1,3-GT pseudogenes, and 81% and 77%, respectively, similar to the murine alpha1,3-GT gene. Thus, the alpha1,3-GT genes and pseudogenes of mammals are highly similar. Ratios of non-synonymous nucleotide changes among the primate pseudogenes as well as the primate genes are still higher than the ratios of non-primates, suggesting that the primate alpha1,3-GT genes tend to be divergent. PMID- 16217799 TI - Expression of ameloblastin during enamel formation in a crocodile. AB - Ameloblastin is an enamel-specific protein that plays critical roles in enamel formation, as well as adhesion between ameloblasts and the enamel matrix, as shown by analyses of ameloblastin-null mice. In the present study, we produced two distinct antibodies that recognize the N-terminus and C-terminus regions of caiman ameloblastin, in order to elucidate the fate of ameloblastin peptides during tooth development. An immunohistochemical study using the antibodies showed that caiman ameloblastin was a tooth-specific matrix protein that may initially be cleaved into two groups, N- and C-terminal peptides, as shown in mammals. The distribution of the N-terminal peptides was much different from that of the C-terminal peptides during enamel formation; however, it was similar to that of mammalian ameloblastin. Although ameloblastin is thought to have a relationship with the enamel prismatic structure in mammals, in the caiman, which has non-prismatic enamel, functional ameloblastin has no relationship with any enamel structure. Consequently, it is suggested that ameloblastin has kept its original functions during the evolutionary transition from reptiles to mammals and that it has been conserved in both lineages during more than 200 million years of evolution. Our results support the notion that ameloblastin acts as a factor for ameloblast adhesion to enamel matrix, because distribution of the C terminal peptides was consistently restricted on the surface layers of enamel matrix specimens ranging from immature to nearly completely mature. The principal molecules that provide the adhesive function are presumably C-terminal peptides. PMID- 16217800 TI - Effects of Low pH acute exposure on survival and gill morphology in Triturus italicus larvae. AB - We examined, from a morphological and ultrastructural point of view, the gill epithelium of Triturus italicus, both in basal conditions and after acute exposure to low pH. Our analysis of gill morphology began with the aim of determining lethal pH levels; we found that the pH value at which 50% of mortality occurs (LC50) is 4.0. We then investigated the effects of the larvae's exposure to a critical value of pH (pH 4.5). No change was observed in the gill during the first 24 hr. After 48 hr, a cellular response was evident and the gills appeared covered with a dense mucous layer. Observations that were carried out by light microscopy (LM) and electron microscopy, both scanning (SEM) and transmission (TEM), showed considerable histological and ultrastructural changes. As regards the principal filament, the alterations resulted in the presence of an external keratinized layer. More changes affected the mitochondria-rich cells (MRCs) in both a qualitative and quantitative way; this cell type shows a wide surface and seems to protrude more than the near pavement cells that, in normal conditions, partially cover the MRCs, thus reducing their external surface. The microvilli were extremely lengthened and often anatomized each other. Changes in the secondary filament affected the thickness of the epithelium, which appeared considerably smaller in the gills of the newts exposed to acid stress. The ciliated cells appeared to be more numerous than in the control specimens and the MRCs showed a widening of the apical surface. PMID- 16217801 TI - Reversal of photoschedule in spring does not prevent photorefractoriness in Siberian hamsters. AB - We studied the influence of light-dark (L:D) cycle reversal on daily variations in the brown adipose tissue (BAT) capacity for nonshivering thermogenesis (NST) in Siberian hamsters (Phodopus sungorus). Continuous and simultaneous measurements of BAT temperature (T(BAT)) and preferred ambient temperature (PT(a)) were made after noradrenaline (NA) injections administered every 4 hr. First, hamsters were acclimated for 4 weeks to an ambient temperature (T(a)) of 23 degrees C and 12L:12D, and then to a reversed photoschedule 12D:12L for 8 weeks. The same was done after a 4- and 8-week acclimation period at the same T(a). We found that after photoschedule reversal, the re-entrainment of T(BAT) and PT(a) rhythms preceded re-entrainment of the NST rhythm. The daily rhythms of T(BAT) and PT(a) were fully re-entrained after 4 weeks of acclimation to the reversed photoschedule, but rhythmicity of the response to NA disappeared. This rhythm was restored in hamsters acclimated to a reversed photoschedule for 8 weeks. We suggest that the daily rhythm of NST capacity is not responsible for generating the rhythm of body temperature (T(b)). Rather, it is a result of the daily rhythm of T(b), but adjusts to the new environment more slowly than the T(b) rhythm. When a daily rhythm of NST was present, the increase in T(BAT) after NA injection was inversely correlated with the pre-injection T(BAT). In addition, NA-induced changes in PT(a) reflected the intensity of NST in BAT; namely, increased T(BAT) was correlated with the post-injection decrease in PT(a). When the increase in T(BAT) was large, animals chose a lower T(a) to dissipate excessive heat and prevent overheating. In the course of the experiments, we recorded a decreased mean NST capacity and increased body mass of hamsters. These changes are representative of the time of photorefractoriness and a transition to a summer status. Despite prolonged exposure to an intermediate day length (12 hr of light) and photoschedule reversal, hamsters continued to change towards their summer condition and were able to acclimate to the new D:L cycle. PMID- 16217802 TI - Triploidy in rainbow trout determined by computer-assisted analysis. AB - This study was designed to assess the use of a computer-assisted system based on erythrocyte measurements as a possible alternative to flow cytometry for identifying triploid rainbow trout (Oncorhynchus mykiss). Blood smears were prepared from 26 triploid and 26 diploid specimens, as determined by flow cytometry after staining blood cells with propidium iodide. The cell and nucleus lengths of 10 erythrocytes were determined in each fish. This was followed by discriminatory analysis to distinguish between diploids and triploids based on their score profiles. Triploid trout showed significantly larger erythrocyte cell and nucleus measurements than their diploid counterparts (N=52; P<0.0001). Erythrocyte length correctly identified 100% of the fish specimens as diploid or triploid, while nucleus length was a less accurate predictor of the level of ploidy. Our findings validate the potential use of computer-assisted analysis for this purpose. PMID- 16217803 TI - Germ cell death in the testis and its relation to spermatogenesis in the wax moth, Galleria mellonella (Lepidoptera: Pyralidae), effects of facultative diapause. AB - The dichotomous spermatogenesis of many Lepidopterans results in the production of two types of sperm: eupyrene sperm possessing a cell nucleus which participates in fertilisation, and apyrene ones, which lose their nuclei during development and whose function remains a mystery. The goal of our study was to analyse spermatogenesis at the end of the larval development of the wax moth, Galleria mellonella, at an optimal temperature of 30 degrees C as well as to describe how they are affected by diapause brought on by a reduction of temperature to 18 degrees C. Spermatogenesis in non-diapausing insects did not differ significantly from that described in other species of Lepidoptera, and any differences found were compared against available literature. Based on the results presented, it may be unequivocally stated that changes in spermatogenesis occur in diapause caused by a suboptimal temperature of 18 degrees C. The main effect of diapause observed in the testes is the degeneration of germ cells, immediately following their differentiation from bipotential spermatocytes. Eupyrene cells seem to reach a more advanced stage of development. Due to the absence of secondary eupyrene spermatocytes in the testis of diapausing insects, it may be surmised that the meiotic divisions, which lead to the formation of secondary spermatocytes and eventually spermatids, do not occur, or are somehow altered. Lastly, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) analyses we performed show that the degenerative changes of eupyrene cells are apoptotic in character. PMID- 16217804 TI - Pigmented epithelium to retinal transdifferentiation and Pax6 expression in larval Xenopus laevis. AB - This study examines the retinal transdifferentiation (TD) of retinal pigmented epithelium (RPE) fragments dissected from Xenopus laevis larvae and implanted into the vitreous chamber of non-lentectomized host eyes. In these experimental conditions, most RPE implants transformed into polarized vesicles in which the side adjacent to the lens maintained the RPE phenotype, while the side adjacent to the host retina transformed into a laminar retina with the photoreceptor layer facing the cavity of the vesicle and with the ganglionar cell layer facing the host retina. The formation of a new retina with a laminar organization is the result of depigmentation, proliferation and differentiation of progenitor cells under the influence of inductive factors from the host retina. The phases of the TD process were followed using BrdU labelling as a marker of the proliferation phase and using a monoclonal antibody (mAbHP1) as a definitive indicator of retina formation. Pigmented RPE cells do not express Pax6. In the early phase of RPE to retinal TD, all depigmented and proliferating progenitor cells expressed Pax6. Changes in the Pax6 expression pattern became apparent in the early phase of differentiation, when Pax6 expression decreased in the presumptive outer nuclear layer (ONL) of the new-forming retina. Finally, during the late differentiation phase, the ONL, which contains photoreceptors, no longer expressed Pax6, Pax6 expression being confined to the ganglion cell layer and the inner nuclear layer. These results indicate that Pax6 may have different roles during the different phases of RPE to retinal TD, acting as an early retinal determinant and later directing progenitor cell fate. PMID- 16217805 TI - Limitation of size by hypoxia in the fruit fly Drosophila melanogaster. AB - The size of an organism is of fundamental importance in all biological processes. It dictates many of the critical interactions and physical factors that delimit the envelope within which an organism can grow. We investigated the effects of reduced oxygen on size and development in the fruit fly Drosophila melanogaster, and showed that limiting the oxygen in the environment limits both whole animal and cell size. When oxygen levels were reduced from 20% in nitrogen to 15%, 10% and 7.5%, there was a linear decrease in both male and female mass. Both cell size and cell number decreased in low oxygen, but changes in cell size accounted for a larger proportion of the overall change in fly size. Cell numbers decreased by a maximum of 11% between flies reared in 20% oxygen and those reared in 7.5% oxygen, whereas cell surface area decreased by 17%. Low oxygen levels increased development time and mortality, but reduced fecundity. Reducing the level of oxygen available significantly slowed development times, with flies reared in 10% oxygen emerging on average 1.5 days later than those in 20% oxygen. The effect of oxygen on size is reversible during embryonic and larval development up to the pupal stage, when final size is set. PMID- 16217806 TI - Effects of prolonged acclimation to intermediate photoperiod and photo-schedule reversal in photosensitive golden hamsters. AB - We investigated the effect of prolonged acclimation to 12 hr of light and photo schedule reversal during the time of photosensitivity in golden hamsters (Mesocricetus auratus). Before the experiments, animals were housed under natural photoperiod and then transferred to 12L:12D (light 12 hr:dark 12 hr) in autumn for 12 weeks. After 4 weeks of acclimation, photo-schedule was reversed (12D:12L). First experiments were done after 4 weeks of acclimation to an ambient temperature (T(a)) of 23 degrees C and a 12L:12D photo-schedule. We examined the daily variations in brown adipose tissue (BAT) capacity for nonshivering thermogenesis (NST). Noradrenaline (NA) injections were given every 4 hr while BAT temperature (T(BAT)) and preferred ambient temperature (PT(a)) were monitored continuously and simultaneously in a thermal gradient system. Then, we investigated the effect of light-dark cycle reversal on a daily rhythm of NST. The hamsters were acclimated to the photo-schedule reversed by 12 hr and the same T(a). After 4 and 8 weeks of acclimation to a reversed photo-schedule, the experiments were repeated. We found that the daily rhythm of the response to NA was entrained to the new light-dark cycle after 4 weeks of acclimation to a reversed photo-schedule. Maximum effect of NA was always recorded during the light phase and in the latter part of the dark phase of the day. NA-induced increase in T(BAT) was correlated with the decrease in PT(a), and was also inversely correlated with pre-injection T(BAT). These data imply that the daily rhythm of the capacity for NST opposes the daily rhythm of body temperature (T(b)). After 8 weeks of acclimation to the reversed photo-schedule, the rhythmicity of the response to NA disappeared, and the daily fluctuations in T(BAT) were the smallest. This lack of rhythm may be a physiological adaptation to winter conditions when the daily amplitude of T(b) rhythm is markedly reduced and, as a consequence, NST capacity does not vary within the day. Moreover, after 8 weeks of acclimation to reversed photo-schedule, NST capacity decreased while response to saline increased. During the experiments, hamsters were photosensitive and were changing to their winter status. However, because of the lack of cold during acclimation, the capacity for NST did not increase. Increased responsiveness to saline, indicating an increase in stress-induced thermogenesis, might be advantageous for "fight or flight" reaction. PMID- 16217807 TI - Adaptation of a nematode parasite to living within the mammalian epithelium. AB - Trichuris muris is a large metazoan pathogen that has been proposed to live intracellularly within living host intestinal epithelial cells. We sought to determine how Trichuris bores its way through the mucosal epithelium and to elucidate the parasite strategies for taking advantage of this intracellular niche. Since the apical surface of the mucosal epithelium is stabilized by the actin cytoskeleton and cell junctions, it remains intact over the worm following its entry into cells. In contrast, non-stabilized lateral membranes of the host epithelial cells are ruptured and cells are killed to form an inert syncytial tunnel. The ventral surface of the nematode worm is studded by pores that overlie bacillary cells; these pores penetrate through the cuticle and are in direct contact with host cytoplasm. From scanning electron micrographs of isolated worms, we calculate that each adult contains approximately 50,000 bacillary cells. The apical surface of the bacillary cells is extensively folded into plicae 40 nm in diameter, thereby increasing the surface area many-fold. Bacillary cells lack organelles for enzyme synthesis and secretion and fail to export protons. However, by confocal light microscopy it was observed that fluorescent macromolecules in excess of 100,000 Da can penetrate into the pores. Taken together, we conclude that the bacillary cells are essential for living inside host epithelium and function predominantly in absorption of soluble molecules from the host mucosal cytoplasm, in essence behaving as an external gut epithelium that is protected from abrasion by the cuticle that surrounds the openings of the bacillary cells. PMID- 16217808 TI - Stressed mothers lay eggs with high corticosterone levels which produce low quality offspring. AB - Organisms frequently encounter stressful ecological conditions. In vertebrates, a major mechanism of physiological response to stress is mediated by the hypothalamic-pituitary-adrenal axis and results in increased secretion of glucocorticosteroids, which can have adverse consequences on diverse phenotypic traits affecting fitness. Maternal stress may thus have carry-over effects on progeny if it influences pre-natal offspring environment in terms of glucocorticosteroid concentration, although this hypothesis has never been tested in any species under field conditions. We manipulated stress experienced by female barn swallows Hirundo rustica, by exposing them to a predator during laying and measured egg corticosterone concentration. Stressed females laid eggs with greater corticosterone concentration than controls exposed to a herbivore. In another experiment, we injected physiological doses of corticosterone in the egg albumen and compared the phenotype of offspring originating from these eggs with their control siblings originating from either sham-inoculated or unmanipulated eggs and reared in the same nest. Eggs injected with corticosterone had lower hatchability and produced fledglings with smaller body size and slower plumage development than did control eggs. Nestling body size in our study population predicts long-term survival. Thus, maternal stress impaired offspring phenotype and viability by increasing transmission of glucocorticosteroids to the eggs. This study identifies a novel mechanism mediating early maternal effects whereby maternal stress affects offspring quality. These results are relevant to biological conservation because they disclose a mechanism that can link environmental conditions to population productivity and viability. PMID- 16217809 TI - Neurobiological specializations in echolocating bats. AB - Although the bat's nervous system follows the general mammalian plan in both its structure and function, it has undergone a number of modifications associated with flight and echolocation. The most obvious neuroanatomical specializations are seen in the cochleas of certain species of bats and in the lower brainstem auditory pathways of all microchiroptera. This article is a review of peripheral and central auditory neuroanatomical specializations in echolocating bats. Findings show that although the structural features of the central nervous system of echolocating microchiropteran bats are basically the same as those of more generalized mammals, certain pathways, mainly those having to do with accurate processing of temporal information and auditory control of motor activity, are hypertrophied and/or organized somewhat differently from those same pathways in nonecholocating species. Through the resulting changes in strengths and timing of synaptic inputs to neurons in these pathways, bats have optimized the mechanisms for analysis of complex sound patterns to derive accurate information about objects in their environment and direct behavior toward those objects. PMID- 16217810 TI - Impact of structure and morphology on charge transport in semiconducting oligomeric thin-film devices. AB - We investigated various thin-film morphologies of vacuum-deposited highly luminescent 2,5-di-n-octyloxy-1,4-bis[4'-(styryl)styryl]benzene (Ooct-OPV5) in a typical light-emitting-diode device structure. Important modifications in the thin-film morphology and structure were obtained by changing the substrate temperature in the range 23-90 degrees C. Structural analysis by X-ray and electron diffraction provided clear evidence for polymorphism in evaporated thin films of Ooct-OPV5. Concomitantly, the hole mobility in the corresponding devices was determined by transient electroluminescence measurements. We demonstrate that the substrate temperature T(sub) is a key parameter that controls the hole mobility of the devices. Increasing T(sub) between 23 and 84 degrees C results in a progressive increase of the zero-field hole mobility from 10(-6) to 10(-4) cm(2) V(-1) s(-1). The increase in hole mobility is correlated to the average grain size in the thin films. In addition, we give evidence for the existence of a peculiar growth mode in the bulk crystal structure of Ooct-OPV5, whereby the (a,b) and (b,c) planes can grow in a homoepitaxial manner. PMID- 16217811 TI - Influence of molecular order on the local work function of nanographene architectures: a Kelvin-probe force microscopy study. AB - We report a Kelvin-probe force microscopy (KPFM) investigation on the structural and electronic properties of different submicron-scale supramolecular architectures of a synthetic nanographene, including extended layers, percolated networks and broken patterns grown from solutions at surfaces. This study made it possible to determine the local work function (WF) of the different pi-conjugated nanostructures adsorbed on mica with a resolution below 10 nm and 0.05 eV. It revealed that the WF strongly depends on the local molecular order at the surface, in particular on the delocalization of electrons in the pi-states, on the molecular orientation at surfaces, on the molecular packing density, on the presence of defects in the film and on the different conformations of the aliphatic peripheral chains that might cover the conjugated core. These results were confirmed by comparing the KPFM-estimated local WF of layers supported on mica, where the molecules are preferentially packed edge-on on the substrate, with the ultraviolet photoelectron spectroscopy microscopically measured WF of layers adsorbed on graphite, where the molecules should tend to assemble face-on at the surface. It appears that local WF studies are of paramount importance for understanding the electronic properties of active organic nanostructures, being therefore fundamental for the building of high-performance organic electronic devices, including field-effect transistors, light-emitting diodes and solar cells. PMID- 16217812 TI - Direct measurement of the end-to-end distance of individual polyfluorene polymer chains. AB - Wide-field imaging of individual multichromophoric molecules and successive photobleaching were used to determine, accurately, the relative position of the chromophores in such systems. First, a polyphenylene dendrimer with well-defined geometry was used to establish the accuracy in localization that can be obtained by this methodology. For a signal-to-noise ratio of 20, interchromophoric distances could be measured with 4 nm accuracy. Next, the method was used to determine the end-to-end distribution of an end-capped polyfluorene polymer. From comparison between the experimental and simulated distributions, information on the conformation of the polymer could be deduced. It was found that the polymer has a nonlinear conformation. A conjugation length of six monomer units gave the best fit of the experimental data to the proposed model. PMID- 16217813 TI - Cooperative photochemical reaction mechanism of femtosecond laser-induced photocoloration in spirooxazine microcrystals. AB - Microcrystalline powders of spirooxazine and spiropyran compounds do not show photocoloration under steady-state illumination, whereas they undergo photochromism on intense femtosecond laser-pulse excitation. We investigated the characteristic mechanism of the crystalline photochromism by studying the photocoloration of spironaphthooxazine (SNO) and its chloro-substituted derivative (Cl-SNO) with our femtosecond diffuse-reflectance spectroscopic system. In particular, femtosecond double-pulse excitation using 390+780-nm pulses and 390+390-nm pulses, with a variable time interval between the two pulses, was applied to reveal an intermediate species involved in the photocoloration. Although 780-nm excitation of an intermediate produced by 390-nm excitation did not lead to isomerization, the 390+390-nm excitation resulted in photocoloration. The yield for SNO decreased on increasing the interval from 40 ps to 5 ns, while that for Cl-SNO was constant. The photocoloration mechanism in the crystalline phase is considered from the viewpoint of the time-dependent density of short-lived transient species, and it is concluded that cooperative interactions of excited states and nonplanar open forms play an important role in femtosecond laser-induced photochromism in these crystals. PMID- 16217814 TI - Circadian variations in the pharmacokinetics, tissue distribution and urinary excretion of nifedipine after a single oral administration to rats. AB - Circadian variations in the pharmacokinetics, tissue distribution and urinary excretion of nifedipine were examined in fasted rats after administering a single oral dose at three different dosing times (08:00 am, 16:00 pm, 00:00 am). The plasma concentrations, the areas under the plasma concentration-time curve from zero to 6 h (AUC(0-6 h)) and the peak plasma concentration (C(max)) were significantly higher in the rats dosed at 08:00 am (immediately inactive), and was lower at 16:00 pm (most inactive) and 00:00 am (most active). The time to reach the C(max) (T(max)) was the shortest in the rats dosed at 08:00 am. It was very interesting to observe the double peak phenomena in the plasma concentration profiles, showing a larger peak followed by a smaller peak. There was a dosing time dependency on the tissue distribution 30 min after administration, showing a similar tendency to the pharmacokinetic behavior. However, there was no distinct dosing time dependency observed at 2 h after administration due to the extensive disposition. The cumulative urine excretion of nifedipine in the rats dosed at 08:00 am was significantly higher (about two-fold) than in those dosed at 16:00 pm and 00:00 am. The pharmacokinetics of nifedipine in the rats was consistent with that observed in human subjects in terms of the day-night clock time but the biological time was the opposite, as marked by the rest-activity cycles. These results may help to explain the circadian time-dependency of nifedipine pharmacokinetics. PMID- 16217815 TI - Structural evidence of mechanical shuttling in condensed monolayers of bistable rotaxane molecules. PMID- 16217816 TI - Accumulation of an E,E,E-triene by the monensin-producing polyketide synthase when oxidative cyclization is blocked. PMID- 16217817 TI - Supramolecular approaches to generate libraries of chelating bidentate ligands for homogeneous catalysis. AB - The process of catalyst discovery and development relying on combinatorial methods has suffered so far from the difficult access to structurally diverse and large libraries of ligands, in particular the structurally more complex class of bidentate ligands. A completely new approach to streamline the difficult ligand synthesis process is to use structurally less complex monodentate ligands that self-assemble in the coordination sphere of a metal center through noncovalent attractive ligand-ligand interactions to generate bidentate, chelating ligands. When complementary attractive ligand-ligand interactions are employed, it is even possible to generate libraries of defined chelate-ligand catalysts by simply mixing two different monomeric ligands. This Minireview summarizes the first approaches and results in this new field of combinatorial homogeneous catalysis. PMID- 16217818 TI - A diiron center stabilized by a bis-TPA ligand as a model of soluble methane monooxygenase: predominant alkene epoxidation with H2O2. PMID- 16217819 TI - A "through-shell" binding isotope effect. PMID- 16217820 TI - Systematic investigation of the Escherichia coli metabolome for the biosynthetic origin of an isocyanide carbon atom. PMID- 16217822 TI - Banana-shaped oligo(aryleneethynylene)s: synthesis and light-emitting characteristics. PMID- 16217821 TI - Chemo-, regio-, and stereoselective cobalt-mediated [2+2+2] cycloaddition of alkynyl boronates to alkenes: 1,3- and 1,4-diboryl-1,3-cyclohexadienes. PMID- 16217823 TI - Janus-faced aluminum: a demonstration of unique Lewis acid and Lewis base behavior of the aluminum atom in [LALB(C6F5)3]. PMID- 16217824 TI - Experimental and theoretical investigation of the electronic and geometrical structures of the Au32 cluster. PMID- 16217825 TI - Ordered multistep synthesis in a single solution directed by DNA templates. PMID- 16217826 TI - Enantiomeric separation of ketoconazole and terconazole antifungals by electrokinetic chromatography: Rapid quantitative analysis of ketoconazole in pharmaceutical formulations. AB - EKC using a neutral CD as chiral selector was applied in this work to the development of a method enabling the enantiomeric separation of ketoconazole and terconazole antifungals. The influence of different experimental conditions such as temperature, CD concentration, pH, and nature and concentration of the buffer on the enantiomeric resolution of the compounds studied was investigated. The use of 10 mM heptakis-(2,3,6-tri-O-methyl)-beta-CD in a 100 mM phosphate buffer (pH 3.5) with a temperature of 15 degrees C allowed the separation of the enantiomers of ketoconazole and terconazole with high resolution (R(s) > 2.0). The rapid separation of ketoconazole enantiomers with an analysis time less than 3 min was carried out after fitting some experimental parameters. The developed method was applied to the determination of ketoconazole in different pharmaceutical formulations. PMID- 16217827 TI - Intracellular FITC-derivatization with PEG. AB - In order to investigate the amino acids (AAs) in plant cells, we explore an avenue for intracellular derivatization with FITC. In this method, FITC was used to mark AAs in living protoplasts derived from embryogenic calli of common wheat (Triticum aestivum L. c.v. Jinan 177) mediated by PEG. After FITC-derivatization, the AAs in the lysate were determined by CE. The result reveals that this PEG method can be used to transfer FITC into plant cells efficiently, which provides a good method for AA analysis in plant cells. PMID- 16217828 TI - Chiral separation of halogenated amino acids by ligand-exchange capillary electrophoresis. AB - The chiral separation of halogenated amino acids by ligand-exchange CE is described. Halogenated amino acids attracted increasing interest in recent years because of their physiological activities. Different chiral selectors, as there are L-4-hydroxyproline, L-histidine, and N-alkyl derivatives of L-4 hydroxyproline in form of their copper(II) complexes, are compared for their chiral recognition ability for halogenated amino acids. The influence of various parameters, such as selector concentration, pH, organic modifier, and field strength, on the resolution was investigated. All halogenated amino acids investigated were baseline-separated under optimized conditions. PMID- 16217830 TI - Chiral separation in capillary electrophoresis using dual neutral cyclodextrins: theoretical models of electrophoretic mobility difference and separation selectivity. AB - Simple equations and theoretical models, related to enantioselectivity (kappa) and C, have been developed for prediction of electrophoretic mobility difference (Deltamu) and separation selectivity (alpha) for enantiomers in CE using dual CDs, where alpha and kappa are defined as the ratio of mu and the ratio of binding constant (K) for enantiomers to each CD, respectively, C the CD concentration, and the average K for enantiomers and each CD. Experiments were carried out using dual CDs as beta-CD and dimethyl-beta-cyclodextrin (DM-beta-CD) and test analytes as five pairs of amphetamine drug enantiomers. A change in observed Deltamu and alpha of enantiomers in dual CDs was found to be in excellent agreement with the theoretical models. For example, in comparison with single CD1, dual CDs can enhance Deltamu and alpha up to the maximum value when enantiomers migrate with the same order in CD1 and CD2, and have the value of rho > 1.0, where rho is the enantioselectivity ratio for CD2 to CD1, while worse Deltamu and alpha are obtained for enantiomers with rho < 1.0. PMID- 16217829 TI - Polymeric alkenoxy amino acid surfactants: IV. effects of hydrophobic chain length and degree of polymerization of molecular micelles on chiral separation of beta-blockers. AB - Four alkenoxy leucine-based surfactants with C8-C11 chains containing a terminal double bond, and one C11 chain surfactant with a terminal triple bond are synthesized and characterized in monomeric and polymeric forms. These polymeric pseudophases are then utilized to study the influence of chain length and DP for the enantioseparations of seven beta-blockers in MEKC. Variations in chain length and concentration of polymeric surfactants showed significant effects on the chiral resolution (Rs) and efficiency (N). A relatively large elution range combined with the highest polarity and aggregation number (A) but the lowest retention time, partial specific volume, and optical rotation generated with C8 polymeric surfactant results in simultaneous enantioseparation of all seven beta blockers with higher N and R(s). In particular, highly hydrophobic beta-blockers are better resolved with shorter hydrocarbon chain even at higher surfactant concentration, which is unachievable with longer chain surfactant. On the other hand, polymer derived from C11-triple bond provided smaller A value compared to C11-double bond surfactant. However, chiral Rs of hydrophobic beta-blockers are still achievable with the C11-triple bond surfactant with enhanced N and shorter analysis time. In addition, effect of polymerization concentration is evaluated by polymerizing all five surfactants at five times their respective CMCs and 100 mM equivalent monomer concentrations. Polymerization of shorter chain (C8 and C9) double-bonded surfactants at five times their respective CMCs results in higher A values with better chiral Rs and N compared to the same two surfactants polymerized at 100 mM. PMID- 16217831 TI - Enantioseparation of chiral N-imidazole derivatives by electrokinetic chromatography using highly sulfated cyclodextrins: mechanism of enantioselective recognition. AB - Baseline separation of ten new substituted [1-(imidazo-1-yl)-1-phenylmethyl)] benzothiazolinone and benzoxazolinone derivatives, with one chiral center, was achieved by CD-EKC using highly sulfated CDs (alpha, beta, gamma highly S-CDs) as chiral selectors. The influence of the type and concentration of the chiral selectors on the enantioseparations was investigated. The highly S-CDs exhibit a very high enantioselectivity power since they allow excellent enantiomeric resolutions compared to those obtained with the neutral CDs. The enantiomers were resolved with analysis times inferior to 2.5 min and resolution factors R(s) of 3.73, 3.90, 1.40, and 4.35 for compounds 1, 2, 3, and 5, respectively, using 25 mM phosphate buffer at pH 2.5 containing either highly S-alpha-CD, highly S-beta CD, and highly S-gamma-CD (3 or 4% w/v) at 298 K, with an applied field of 0.30 kV/cm. The determination of the enantiomer migration order for the various analytes and the study of the analyte structure-enantioseparation relationships display the high contribution of the interactions between the analytes phenyl ring and the CDs to the enantiorecognition process. The thermodynamic study of the analyte-CD affinities permits us to improve our knowledge about the enantioseparation mechanism. PMID- 16217832 TI - Enantioselective separation of the sunscreen agent 3-(4-methylbenzylidene) camphor by electrokinetic chromatography: Quantitative analysis in cosmetic formulations. AB - 3-(4-Methylbenzylidene)-camphor (MBC) is a chiral sunscreen agent used in cosmetic products. In this work, the enantioseparation of MBC has been performed by EKC and applied to the analysis of the MBC enantiomers in cosmetic creams. Different experimental conditions (type and concentration of the chiral selector, temperature, and sample solvent) have been optimized. Due to the neutral nature of this compound, anionic CD derivatives were investigated as chiral selectors. Carboxymethylated-beta-CD (CM-beta-CD) showed the highest chiral separation power, observing that a 15 mM concentration of this CD at a working temperature of 15 degrees C enabled to obtain the highest enantioresolution. However, under these conditions, tailing of peaks obtained for the enantiomers was observed. The addition of increasing concentrations of the neutral alpha-CD to CM-beta-CD at a 15 mM concentration in a 100 mM borate buffer at pH 9.0 improved the enantiomeric separation and decreased peak tailing. The use of DMF for the total dissolution of the cosmetic creams, and methanol:water (1:1 v/v) for appropriate dilution enabled to observe good shape and size for the peaks of the MBC enantiomers. After optimizing a method for the preconditioning of the capillary, the analytical characteristics of the chiral separation method for the analysis of MBC were investigated. Linearity, LODs and LOQs, precision (instrumental repeatability, method repeatability, intermediate precision), accuracy, and selectivity were evaluated. The method was applied to analyze MBC enantiomers contained in two commercial cosmetic creams containing racemic MBC and to study the skin absorption of this compound with time. PMID- 16217833 TI - Chiral analysis of pollutants and their metabolites by capillary electromigration methods. AB - Chiral separation of enantiomers is one of the most challenging tasks for any analytical technique including CE. Since the first report in 1985 showing the great possibilities of CE for the separation of chiral compounds, the amount of publications concerning this topic has quickly increased. Although chiral electromigration methods have mainly been used for enantioseparation of drugs and pharmaceuticals, they have also been applied to analyze chiral pollutants. This article intends to provide an updated overview, including works published till January 2005, on the principal applications of CE to the chiral analysis of pollutants and their metabolites, with special emphasis on articles published in the last 10 years. The main advantages and drawbacks regarding the use of CE for chiral separation of pollutants are addressed including some discussion on the foreseen trends of electromigration procedures applied to chiral analysis of contaminants. PMID- 16217834 TI - Enantioseparation of phenothiazines in cyclodextrin-modified capillary zone electrophoresis using sulfated cyclodextrins as chiral selectors. AB - In this study, enantioseparations of five phenothiazines, including promethazine, ethopropazine, trimeprazine, methotrimeprazine, and thioridazine, in CD-modified CZE using dual CD systems consisting of randomly sulfate-substituted CD (MI-S beta-CD) and a neutral CD as chiral selectors in a citrate buffer (100 mM) at pH 3.0 were investigated. The results indicate that MI-S-beta-CD is an excellent chiral selector for enantioseparation of ethopropazine. The enantiomers of promethazine can also be baseline-resolved with MI-S-beta-CD at concentrations in the range of 0.5-1.0% w/v. On the other hand, thioridazine and trimeprazine interact strongly with neutral CDs. As a result, the enantioselectivity of these two phenothiazines is remarkably and synergistically enhanced with increasing the concentration of neutral CDs in the presence of MI-S-beta-CD and simultaneous enantioseparations of these phenothiazines, except for methotrimeprazine, could favorably be achieved with the use of dual CD systems. Moreover, by varying the concentration of beta-CD or gamma-CD at a fixed concentration of MI-S-beta-CD (0.75% w/v) reversal of the enantiomer migration order of promethazine occurred. This may be attributable to the opposite effects of charged and neutral CDs on the mobility of the enantiomers of promethazine. PMID- 16217835 TI - Confidence intervals for an effect size measure based on the Mann-Whitney statistic. Part 2: asymptotic methods and evaluation. AB - Several asymptotic confidence interval methods for U/mn, the Mann-Whitney U statistic divided by the product of the two sample sizes, are developed and evaluated alongside published methods. Novel methods derived from Gaussian and beta models perform well, though overall a modification of the Hanley-McNeil approach (method 5 in this evaluation) performed better and is recommended for calculating confidence intervals for U/mn for continuously distributed data. PMID- 16217836 TI - Bayesian multivariate hierarchical transformation models for ROC analysis. AB - A Bayesian multivariate hierarchical transformation model (BMHTM) is developed for receiver operating characteristic (ROC) curve analysis based on clustered continuous diagnostic outcome data with covariates. Two special features of this model are that it incorporates non-linear monotone transformations of the outcomes and that multiple correlated outcomes may be analysed. The mean, variance, and transformation components are all modelled parametrically, enabling a wide range of inferences. The general framework is illustrated by focusing on two problems: (1) analysis of the diagnostic accuracy of a covariate-dependent univariate test outcome requiring a Box-Cox transformation within each cluster to map the test outcomes to a common family of distributions; (2) development of an optimal composite diagnostic test using multivariate clustered outcome data. In the second problem, the composite test is estimated using discriminant function analysis and compared to the test derived from logistic regression analysis where the gold standard is a binary outcome. The proposed methodology is illustrated on prostate cancer biopsy data from a multi-centre clinical trial. PMID- 16217837 TI - Electrospray ionization of nucleic acid aptamer/small molecule complexes for screening aptamer selectivity. AB - Molecular recognition of small molecule ligands by the nucleic acid aptamers for tobramycin, ATP, and FMN has been examined using electrospray ionization mass spectrometry (ESI-MS). Mass spectrometric data for binding stoichiometry and relative binding affinity correlated well with solution data for tobramycin aptamer complexes, in which aptamer/ligand interactions are mediated by hydrogen bonds. For the ATP and FMN aptamers, where ligand interactions involve both hydrogen bonding and significant pi-stacking, the relative binding affinities determined by MS did not fully correlate with results obtained from solution experiments. Some high-affinity aptamer/ligand complexes appeared to be destabilized in the gas phase by internal Coulombic repulsion. In CAD experiments, complexes with a greater number of intermolecular hydrogen bonds exhibited greater gas-phase stability even in cases when solution binding affinities were equivalent. These results indicate that in at least some cases, mass spectrometric data on aptamer/ligand binding affinities should be used in conjunction with complementary techniques to fully assess aptamer molecular recognition properties. PMID- 16217839 TI - Dispersion frailty models and HGLMs. AB - In medical research recurrent event times can be analysed using a frailty model in which the frailties for different individuals are independent and identically distributed. However, such a homogeneous assumption about frailties could sometimes be suspect. For modelling heterogeneity in frailties we describe dispersion frailty models arising from a new class of models, namely hierarchical generalized linear models. Using the kidney infection data we illustrate how to detect and model heterogeneity among frailties. Stratification of frailty models is also investigated. PMID- 16217838 TI - Combination of solid-phase affinity capture on magnetic beads and mass spectrometry to study non-covalent interactions: example of minor groove binding drugs. AB - A simple and novel approach was developed to detect non-covalent interactions. It is based on combination of solid-phase affinity capture with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). One of the interacting molecules is bound to magnetic beads and is incubated with the target molecules in solution. The complex bound on the solid support is removed from the solution and transferred for MALDI analysis. Mass spectrometry is used only to detect the target compound, which is far more straightforward than detecting the intact non-covalent complex. To demonstrate the applicability of the method, an AT-rich oligonucleotide (5'-CCCCCAATTCCCCC-3') and its complementary biotinylated sequence (5'-biotin-GGGGGAATTGGGGG-3') were hybridized and immobilized to paramagnetic particles by streptavidin-biotin interaction. The immobilized duplex oligonucleotide was reacted with minor groove binding drugs, Netropsin, Distamycin A, Hoechst 33258 and 4',6-diamidino-2-phenylindole. The resulting DNA-drug complex bound to the particles was separated and analyzed by linear MALDI-TOFMS after washing. Drugs were selectively detected in the spectra. Relative binding strengths were also estimated using competitive complexation. PMID- 16217840 TI - Power and money in cluster randomized trials: when is it worth measuring a covariate? AB - The power to detect a treatment effect in cluster randomized trials can be increased by increasing the number of clusters. An alternative is to include covariates into the regression model that relates treatment condition to outcome. In this paper, formulae are derived in order to evaluate both strategies on basis of their costs. It is shown that the strategy that uses covariates is more cost efficient in detecting a treatment effect when the costs to measure these covariates are small and the correlation between the covariates and outcome is sufficiently large. The minimum required correlation depends on the cluster size, and the costs to recruit a cluster and to measure the covariate, relative to the costs to recruit a person. Measuring a covariate that varies at the person level only is recommended when cluster sizes are small and the costs to recruit and measure a cluster are large. Measuring a cluster level covariate is recommended when cluster sizes are large and the costs to recruit and measure a cluster are small. An illustrative example shows the use of the formulae in a practical setting. PMID- 16217841 TI - Dichotomizing continuous predictors in multiple regression: a bad idea. AB - In medical research, continuous variables are often converted into categorical variables by grouping values into two or more categories. We consider in detail issues pertaining to creating just two groups, a common approach in clinical research. We argue that the simplicity achieved is gained at a cost; dichotomization may create rather than avoid problems, notably a considerable loss of power and residual confounding. In addition, the use of a data-derived 'optimal' cutpoint leads to serious bias. We illustrate the impact of dichotomization of continuous predictor variables using as a detailed case study a randomized trial in primary biliary cirrhosis. Dichotomization of continuous data is unnecessary for statistical analysis and in particular should not be applied to explanatory variables in regression models. PMID- 16217842 TI - Power and sample size computations in simultaneous tests for non-inferiority based on relative margins. AB - In this paper, we address the problem of calculating power and sample sizes associated with simultaneous tests for non-inferiority. We consider the case of comparing several experimental treatments with an active control. The approach is based on the ratio view, where the common non-inferiority margin is chosen to be some percentage of the mean of the control treatment. Two power definitions in multiple hypothesis testing, namely, complete power and minimal power, are used in the computations. The sample sizes associated with the ratio-based inference are also compared with that of a comparable inference based on the difference of means for various scenarios. It is found that the sample size required for ratio based inferences is smaller than that of difference-based inferences when the relative non-inferiority margin is less than one and when large response values indicate better treatment effects. The results are illustrated with examples. PMID- 16217843 TI - Microscale sample deposition onto hydrophobic target plates for trace level detection of neuropeptides in brain tissue by MALDI-MS. AB - A sample preparation method that combines a modified target plate with a nanoscale reversed-phase column (nanocolumn) was developed for detection of neuropeptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A gold-coated MALDI plate was modified with an octadecanethiol (ODT) self-assembled monolayer to create a hydrophobic surface that could concentrate peptide samples into a approximately 200-500-microm diameter spot. The spot sizes generated were comparable to those obtained for a substrate patterned with 200-microm hydrophilic spots on a hydrophobic substrate. The sample spots on the ODT-coated plate were 100-fold smaller than those formed on an unmodified gold plate with a 1-microl sample and generated 10 to 50 times higher mass sensitivity for peptide standards by MALDI-TOF MS. When the sample was deposited on an ODT-modified plate from a nanocolumn, the detection limit for peptides was as low as 20 pM for 5-microl samples corresponding to 80 amol deposited. This technique was used to analyze extracts of microwave-fixed tissue from rat brain striatum. Ninety-eight putative peptides were detected including several that had masses matching neuropeptides expected in this brain region such as substance P, rimorphin, and neurotensin. Twenty-three peptides had masses that matched peaks detected by capillary liquid chromatography with electrospray ionization MS. PMID- 16217844 TI - Porous polymer monolith assisted electrospray from a glass microdevice. AB - The coupling of a lab-on-a-chip microfluidic device to a nanoelectrospray ionization mass spectrometer has the potential to automate many routine analytical procedures and produce a powerful analytical tool. However, past coupling strategies have relied on complex manufacturing steps including drilling and etching the device to attach a capillary or building a nanospray emitter directly into the device. This study shows that a nanospray emitter can be easily fabricated using a porous polymer monolith (PPM) at the end of a glass microdevice. These devices are able to obtain a stable electrospray at a variety of flow rates (50-500 nL/min) but optimal results are obtained at lower flow rates (50-100 nL/min) compatible with electroosmotic flow processes. The PPM is photo-patterned so that it can be placed in any position within the channel of the device with no dead volume. The porous character and the hydrophobic nature of the PPM both aid in development of a stable electrospray process. Total ion current traces for the constant infusion of leucine-enkephalin and PPG show relative standard errors as low as 4%, and produce mass spectra with good signal to-noise (S/N 43) from only 2 fmol of material. In addition, multiple experiments in a given day show good repeatability with variability as low as 13%, and the multiple flow paths inherent in the PPM limit sprayer clogging. PMID- 16217845 TI - Modelling antigenic drift in weekly flu incidence. AB - Since influenza in humans is a major public health threat, the understanding of its dynamics and evolution, and improved prediction of its epidemics are important aims. Underlying its multi-strain structure is the evolutionary process of antigenic drift whereby epitope mutations give mutant virions a selective advantage. While there is substantial understanding of the molecular mechanisms of antigenic drift, until now there has been no quantitative analysis of this process at the population level. The aim of this study is to develop a predictive model that is of a modest-enough structure to be fitted to time series data on weekly flu incidence. We observe that the rate of antigenic drift is highly non uniform and identify several years where there have been antigenic surges where a new strain substantially increases infective pressure. The SIR-S approach adopted here can also be shown to improve forecasting in comparison to conventional methods. PMID- 16217846 TI - Joint analysis of repeatedly observed continuous and ordinal measures of disease severity. AB - In biomedical studies often multiple measures of disease severity are recorded over time. Although correlated, such measures are frequently analysed separately of one another. Joint analysis of the outcomes variables has several potential advantages over separate analyses. However, models for response variables of different types (discrete and continuous) are challenging to define and to fit. Herein we propose correlated probit models for joint analysis of repeated measurements on ordinal and continuous variables measuring the same underlying disease severity over time. We demonstrate how to rewrite the models so that maximum-likelihood estimation and inference can be performed with standard software. Simulation studies are performed to assess efficiency gains in fitting the responses together rather than separately and to guide response variable selection for future studies. Data from a depression clinical trial are used for illustration. PMID- 16217847 TI - Improving ecological inference using individual-level data. AB - In typical small-area studies of health and environment we wish to make inference on the relationship between individual-level quantities using aggregate, or ecological, data. Such ecological inference is often subject to bias and imprecision, due to the lack of individual-level information in the data. Conversely, individual-level survey data often have insufficient power to study small-area variations in health. Such problems can be reduced by supplementing the aggregate-level data with small samples of data from individuals within the areas, which directly link exposures and outcomes. We outline a hierarchical model framework for estimating individual-level associations using a combination of aggregate and individual data. We perform a comprehensive simulation study, under a variety of realistic conditions, to determine when aggregate data are sufficient for accurate inference, and when we also require individual-level information. Finally, we illustrate the methods in a case study investigating the relationship between limiting long-term illness, ethnicity and income in London. PMID- 16217848 TI - A multiscale method for disease mapping in spatial epidemiology. AB - The effects of spatial scale in disease mapping are well-recognized, in that the information conveyed by such maps varies with scale. Here we provide an inferential framework, in the context of tract count data, for describing the distribution of relative risk simultaneously across a hierarchy of multiple scales. In particular, we offer a multiscale extension of the canonical standardized mortality ratio (SMR), consisting of Bayesian posterior-based strategies for both estimation and characterization of uncertainty. As a result, a hierarchy of informative disease and confidence maps can be produced, without the need to first try to identify a single appropriate scale of analysis. We explore the behaviour of the proposed methodology in a small simulation study, and we illustrate its usage through an application to data on gastric cancer in Tuscany. PMID- 16217849 TI - Forecasting age-specific breast cancer mortality using functional data models. AB - Accurate estimates of future age-specific incidence and mortality are critical for allocation of resources to breast cancer control programmes and evaluation of screening programmes. The purpose of this study is to apply functional data analysis techniques to model age-specific breast cancer mortality time trends, and forecast entire age-specific mortality functions using a state-space approach. We use annual unadjusted breast cancer mortality rates in Australia, from 1921 to 2001 in 5 year age groups (45 to 85+). We use functional data analysis techniques where mortality and incidence are modelled as curves with age as a functional covariate varying by time. Data are smoothed using non-parametric smoothing methods then decomposed (using principal components analysis) to estimate basis functions that represent the functional curve. Period effects from the fitted coefficients are forecast then multiplied by the basis functions, resulting in a forecast mortality curve with prediction intervals. To forecast, we adopt a state-space approach and an automatic modelling framework for selecting among exponential smoothing methods.Overall, breast cancer mortality rates in Australia remained relatively stable from 1960 to the late 1990s, but have declined over the last few years. A set of four basis functions minimized the mean integrated squared forecasting error and account for 99.3 per cent of variation around the mean mortality curve. Twenty year forecasts suggest a continuing decline, but at a slower rate, and stabilizing beyond 2010. Forecasts show a decline in all age groups with the greatest decline in older women. The proposed methods have the potential to incorporate important covariates such as hormone replacement therapy and interventions to represent mammographic screening. This would be particularly useful for evaluating the impact of screening on mortality and incidence from breast cancer. PMID- 16217850 TI - Comparing onset of antidepressant action using a repeated measures approach and a traditional assessment schedule. AB - BACKGROUND: It has been recommended that onset of antidepressant action be assessed using survival analyses with assessments taken at least twice per week. However, such an assessment schedule is problematic to implement. The present study assessed the feasibility of comparing onset of action between treatments using a categorical repeated measures approach with a traditional assessment schedule. METHOD: Four scenarios representative of antidepressant clinical trials were created by varying mean improvements over time. Two assessment schedules were compared within the simulated 8-week studies: (i) 'frequent' assessment--16 postbaseline visits (twice-weekly for 8 weeks); (ii) 'traditional' assessment--5 postbaseline visits (Weeks 1, 2, 4, 6, and 8). Onset was defined as a 20 per cent improvement from baseline, and had to be sustained at all subsequent assessments. Differences between treatments were analysed with a survival analysis (KM = Kaplan-Meier product limit method) and a categorical mixed-effects model repeated measures analysis (MMRM-CAT). RESULTS: More frequent assessments resulted in small reductions in empirical standard errors compared with traditional assessments for both analytic methods. More frequent assessments altered estimates of treatment group differences in KM such that power was increased when the difference between treatments was increasing over time, but power decreased when the treatment difference decreased over time. More frequent assessments had a minimal effect on estimates of treatment group differences in MMRM-CAT. The MMRM-CAT analysis of data from a traditional assessment schedule provided adequate control of type I error, and had power comparable to or greater than that with KM analyses of data from either a frequent or a traditional assessment schedule. CONCLUSION: In the scenarios tested in this study it was reasonable to assess treatment group differences in onset of action with MMRM-CAT and a traditional assessment schedule. Additional research is needed to assess whether these findings hold in data with drop-out and across definitions of onset. PMID- 16217851 TI - Statistical properties and inference of the antimicrobial MIC test. AB - A common method for measuring the drug-specific minimum inhibitory concentration (MIC) of an antibacterial agent is via a two-fold broth dilution test known as the MIC test. Because this procedure implicitly rounds data upward, inference based on unadjusted measurements is biased and overestimates bacterial resistance to a drug. We detail this test procedure and its associated bias, which, in many cases, has an expected value of approximately 0.5 on the log(2) scale. In addition, new bias-corrected estimates of resistance are proposed. A numeric example is used to illustrate the extent to which the traditional resistance estimate can overestimate the true proportion of resistant strains, a phenomenon which is remedied by using the proposed estimates. PMID- 16217852 TI - Adaptation of multiple logistic regression to a multiple inverse sampling design: application to the Isfahan healthy heart program. AB - In observational and experimental studies in the health sciences involving human populations, it is sometimes considered desirable to recruit subjects according to designs that specify a predetermined number of subjects in each of several mutually exclusive classes (generally but not necessarily demographic in nature). This type of adaptive sampling design, now generally referred to as multiple inverse sampling (MIS), has received recent attention, and estimation methods are now available for several sequential MIS sampling designs. In this class of designs, subjects are sampled randomly and sequentially, usually one at a time, until all classes have the pre-specified number of subjects. In this paper, we extend MIS for finite population sampling to estimation of the parameters in multiple logistic regression under MIS. Using estimated logistic regression parameters and cost components obtained from the Isfahan Healthy Heart Program (IHHP), we report findings from a simulation experiment in which it appears that, at fixed cost, MIS at the last stage of sampling compares favourably to simple random sampling. The IHHP is a large community intervention study for prevention of cardiovascular disease being conducted in Isfahan, Iran and two other cities in Iran. The IHHP identified subjects through a multistage sample survey in which MIS was used at the final stage of sampling. MIS is one of several methods of adaptive sampling that are generating considerable interest and show promise of being useful in a wide variety of applications. PMID- 16217853 TI - Generalizing the TITE-CRM to adapt for early- and late-onset toxicities. AB - Due to the staggered entry of subjects in phase I trials, some subjects will only be partially through the study when others are ready to be enrolled. Nonetheless, many phase I designs focus solely upon whether or not subjects experience toxicity, thereby determining the maximum tolerated dose (MTD) with a binomial likelihood using data from fully observed subjects. The time-to-event continual reassessment method (TITE-CRM) was the first attempt to incorporate information from partially observed subjects by using a weighted binomial likelihood in which the weights are based upon the actual toxicity time distribution. Unfortunately, it is difficult to accurately estimate the toxicity time distribution because only a small proportion of enrolled subjects will experience toxicity. Creators of the TITE-CRM propose the simple alternative of weighting subjects by the proportion of time observed, as well as two adaptive weights to adjust for late onset toxicities. As a alternative to these approaches, we suggest assuming the toxicity times, as a proportion of the total time under observation, have a Beta distribution with parameters 1.0 and theta; we also allow theta to vary by dose. The value of theta allows us to reflect the occurrence of early- or late-onset toxicities without correctly specifying the actual distribution of toxicity times. Through this model, we do not necessarily expect to improve identification of the MTD, but rather hope to reduce the exposure of subjects to overly toxic doses. Through simulation, we examine how well our model identifies the MTD and allocates dose assignments in three scenarios investigated by previous publications. PMID- 16217854 TI - Likelihood approaches to the non-parametric two-sample problem for right-censored data. AB - The classical two-sample problem with random right-censoring is considered. We show that non- parametric likelihood techniques can be used to obtain tests for either the identity hypothesis or the non-parametric Behrens-Fisher hypothesis (NBFH). In the case of the identity hypothesis, a special imputed permutation distribution is used to estimate the distribution under the null hypothesis. In the case of the NBFH, simulation from the constrained non-parametric maximum likelihood estimate is used. Simulation shows that the tests using either approximation have excellent control of the type I error rate, even with quite small sample sizes. Further, for Lehmann-type alternatives the likelihood-based methods have similar power to the logrank test, while for the non-Lehmann-type alternatives tried here the likelihood-based methods have superior power. PMID- 16217855 TI - Missing data methods for the assessment of surrogate outcomes and treatment mechanisms in clinical trial substudies. AB - Hypotheses about the mechanisms of action by which a treatment affects a clinical outcome may prompt consideration of an alternative outcome as a potential surrogate. In many cases, due to costs or other factors, the candidate for surrogacy will only be measured for patients randomized to a substudy within the main trial. In this situation, the substudy patients provide information about links between the true and surrogate outcomes and the treatment, and these links can be exploited, using methods for handling missing covariates, to allow available information for patients not in the substudy to be incorporated into the analysis. The increased precision with which the treatment effect can be estimated using these methods, compared with using substudy data alone, in turn allows more precise estimates of measures of surrogacy. This paper reviews and compares some methods for handling missing covariate data and applies the methodology to a large heart attack trial, in order to investigate the properties of four measures for assessing the extent to which early response to thrombolytic therapy, as measured by an improvement in coronary blood flow, can be regarded as a surrogate for 30-day survival following heart attack. Design issues for substudies intended to assess treatment mechanisms are also considered. In particular, we consider how the precision of surrogate measures varies with the size of the substudy relative to the main trial. The results suggest that for reasonable surrogates, substudies substantially smaller than the main study can extract most of the available information regarding surrogacy. PMID- 16217856 TI - An appraisal of methods for the analysis of longitudinal categorical data with MAR drop-outs. AB - A number of methods for analysing longitudinal ordinal categorical data with missing-at-random drop-outs are considered. Two are maximum-likelihood methods (MAXLIK) which employ marginal global odds ratios to model associations. The remainder use weighted or unweighted generalized estimating equations (GEE). Two of the GEE use Cholesky-decomposed standardized residuals to model the association structure, while another three extend methods developed for longitudinal binary data in which the association structures are modelled using either Gaussian estimation, multivariate normal estimating equations or conditional residuals. Simulated data sets were used to discover differences among the methods in terms of biases, variances and convergence rates when the association structure is misspecified. The methods were also applied to a real medical data set. Two of the GEE methods, referred to as Cond and ML-norm in this paper and by their originators, were found to have relatively good convergence rates and mean squared errors for all sample sizes (80, 120, 300) considered, and one more, referred to as MGEE in this paper and by its originators, worked fairly well for all but the smallest sample size, 80. PMID- 16217857 TI - Determination of steroid hormones, hormone conjugates and macrolide antibiotics in influents and effluents of sewage treatment plants utilising high-performance liquid chromatography/tandem mass spectrometry with electrospray and atmospheric pressure chemical ionisation. AB - In this study we present a high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) method which has been elaborated to analyse steroid hormones, hormone conjugates, oral contraceptives and macrolide antibiotics unchanged in unfiltered influents and effluents of sewage treatment plants (STPs). HPLC separation of the steroid hormones was achieved in 35 min, as well as those of the antibiotics. The analytes were extracted by solid-phase extraction, followed by clean-up using size exclusion chromatography (SEC). For the final quantification HPLC/MS/MS was used. The two ionisation modes, electrospray ionisation (ESI) and atmospheric pressure chemical ionisation (APCI), in HPLC/MS/MS were compared for the analysis of steroid hormones. For quantitative results drastic matrix effects were observed while using ESI. These effects were less pronounced while using APCI. These pitfalls were additionally reduced by clean-up using SEC as well as isotope dilution. Additionally, two multiple reaction monitoring (MRM) transitions per compound were used to prevent false positive results. Recovery experiments with spiked tap water with concentrations varying from 1 to 1000 ng/L gave constant recovery rates: The recovery rates for the hormones and conjugates ranged from 58 to 107%, those of the contraceptives ranged from 83 to 109%. The relative standard deviation was found to be 7 to 24% and the limits of detection were 0.1 to 4.5 ng/L. The recovery rates of the macrolide antibiotics ranged from 76 to 103%, while the relative standard deviation was found to be 7 to 14% and the limits of detection ranged from 0.6 to 1.8 ng/L. The maximum concentrations found in influents of a STP was 470 ng/L for estriol and 1200 ng/L for erythromycin. PMID- 16217859 TI - The importance of varying the event generation process in simulation studies of statistical methods for recurrent events. AB - Statistical methods for the analysis of recurrent events are often evaluated in simulation studies. A factor rarely varied in such studies is the underlying event generation process. If the relative performance of statistical methods differs across generation processes, then studies based upon one process may mislead. This paper describes the simulation of recurrent events data using four models of the generation process: Poisson, mixed Poisson, autoregressive, and Weibull. For each model four commonly used statistical methods for the analysis of recurrent events (Cox's proportional hazards method, the Andersen-Gill method, negative binomial regression, the Prentice-Williams-Peterson method) were applied to 200 simulated data sets, and the mean estimates, standard errors, and confidence intervals obtained. All methods performed well for the Poisson process. Otherwise, negative binomial regression only performed well for the mixed Poisson process, as did the Andersen-Gill method with a robust estimate of the standard error. The Prentice-Williams-Peterson method performed well only for the autoregressive and Weibull processes. So the relative performance of statistical methods depended upon the model of event generation used to simulate data. In conclusion, it is important that simulation studies of statistical methods for recurrent events include simulated data sets based upon a range of models for event generation. PMID- 16217860 TI - Decision-theoretic designs for dose-finding clinical trials with multiple outcomes. AB - A decision-theoretic framework is proposed for designing sequential dose-finding trials with multiple outcomes. The optimal strategy is solvable theoretically via backward induction. However, for dose-finding studies involving k doses, the computational complexity is the same as the bandit problem with k-dependent arms, which is computationally prohibitive. We therefore provide two computationally compromised strategies, which is of practical interest as the computational complexity is greatly reduced: one is closely related to the continual reassessment method (CRM), and the other improves CRM and approximates to the optimal strategy better. In particular, we present the framework for phase I/II trials with multiple outcomes. Applications to a pediatric HIV trial and a cancer chemotherapy trial are given to illustrate the proposed approach. Simulation results for the two trials show that the computationally compromised strategy can perform well and appear to be ethical for allocating patients. The proposed framework can provide better approximation to the optimal strategy if more extensive computing is available. PMID- 16217861 TI - Regular exercise eases inflammation. PMID- 16217858 TI - Advantage of using CBA/N strain mice in a non-radioisotopic modification of the local lymph node assay. AB - The murine local lymph node assay (LLNA) is currently recognized as a stand-alone test method for determining the skin sensitizing potential of chemicals. It has been incorporated into the official test guidelines published by some authorities, including the OECD. To avoid the use of radioisotopes, efforts have been made recently to develop non-radioisotopic modifications of the LLNA. A non radioisotopic modification of the LLNA was developed previously using 5-bromo-2' deoxyuridine (BrdU) incorporation (non-RI LLNA). However, the non-RI LLNA was found to be somewhat less sensitive than the standard assay. This study reports the advantage of using mice of the CBA/N strain in the non-RI LLNA to improve the sensitivity of this method. The non-RI LLNA was performed using CBA/JN and CBA/N mice exposed to one of four confirmed skin sensitizers, 2,4-dinitrochlorobenzene (DNCB), eugenol (EG), isoeugenol (IEG) or alpha-hexylcinnamic aldehyde (HCA), and to one non-sensitizer, propylene glycol (PG). The EC3 values for DNCB, IEG, EG, HCA and PG were calculated to be 0.1%, 9.6%, 40.6%, 45.5% and >50% in CBA/JN mice and 0.08%, 1.9%, 10.7%, 20.3% and >50% in CBA/N mice, respectively. The EC3 values for DNCB, IEG, EG, HCA and PG in the standard LLNA using CBA/Ca mice and radioisotopes were reported elsewhere as being 0.08%, 1.3%, 13.0%, 8.0% and >50%, respectively. The EC3 values derived from the CBA/N mice in the non-RI LLNA were nearly equivalent to the EC3 values obtained using the standard radioisotopic LLNA with CBA/Ca mice. These data suggest that the use of CBA/N mice may provide a realistic opportunity to develop a version of the LLNA that does not have a requirement for the use of radioisotopes, but which nevertheless has sensitivity approaching, or comparable to, the standard method. PMID- 16217862 TI - Drug combinations best for heart disease. PMID- 16217863 TI - Limit Natrecor to hospital use, say experts. PMID- 16217864 TI - [Abecedary of the "Annales". Part 6]. AB - The terms included and detailed in the present part are: cost-effectiveness analysis, cost-utility analysis, cost-benefit analysis, proportional hazards (Cox) model, inclusion criteria, endpoints or outcomes. PMID- 16217865 TI - [Strategy for management of ovarian immature teratoma. About three cases and review]. AB - Ovarian immature teratoma is a malignant germ cell tumor and represents less than 1% of ovarian malignant tumors. Tissues are derived from the three germ layers (endo-, meso- and ectoderm). Tumor grading is based on the amount of immature neuroepithelium present. The prognosis is directly correlated to histologic grade. Rapid growth leads to large tumors with an early diagnosis. Therapeutic management is balanced between adjuvant chemotherapy and surgery alone. Nevertheless, as immature teratoma mostly occurs in young women, the main objective is to preserve fertility. PMID- 16217866 TI - Microtubules as a gyroscope for cells on the go. PMID- 16217867 TI - Basal lamina instructs innervation. PMID- 16217868 TI - The opposite of eureka II: nature. By Mole. PMID- 16217869 TI - Quiz page. Acquired kidney disease (ACKD) with associated bilateral renal cell carcinoma. PMID- 16217870 TI - Response and adaptation by plants to flooding stress. AB - Stress on plants imposed by flooding of the soil and deeper submergence constitutes one of the major abiotic constraints on growth, species' distribution and agricultural productivity. Flooding stress is also a strong driver of adaptive evolution. This has resulted in a wide range of biochemical, molecular and morphological adaptations that sanction growth and reproductive success under episodic or permanently flooded conditions that are highly damaging to the majority of plant species. However, even seemingly poorly adapted species possess some short-term resilience that is important for overall success of these plants in various habitats. The papers contained in this Special Issue address these topics and emphasize molecular, biochemical and developmental processes that impact on flooding tolerance. Most of the articles are based on lectures given to the 8th Conference of the International Society for Plant Anaerobiosis (ISPA), held at the University of Western Australia, Perth, 20-24 September, 2004. Reviews and research papers are presented from the leading laboratories currently working on plant responses to flooding stress. PMID- 16217872 TI - Guarding the operatory door. PMID- 16217871 TI - President's message. Advocacy: it may be just your thing. PMID- 16217874 TI - The importance of professional organization as it relates to advocacy. PMID- 16217873 TI - National advocacy. PMID- 16217875 TI - Advocacy for the national examination. PMID- 16217876 TI - Disposal of explanted items. PMID- 16217877 TI - [Remembrance of the first congress of Serbian physicians and naturalists on the 100th anniversary]. PMID- 16217878 TI - Margot Kruskall. PMID- 16217879 TI - Cold, cough, allergy, bronchodilator, and antiasthmatic drug products for over the-counter human use; amendment of final monograph for over-the-counter nasal decongestant drug products. Final rule. AB - The Food and Drug Administration (FDA) is amending the final monograph (FM) for over-the-counter (OTC) nasal decongestant drug products (drug products used to relieve nasal congestion due to a cold, hay fever, or other upper respiratory allergies) to remove the indication "for the temporary relief of nasal congestion associated with sinusitis" and to prohibit use of the terms "sinusitis" and "associated with sinusitis" elsewhere on the labeling. This final rule is part of FDA's ongoing review of OTC drug products. PMID- 16217881 TI - [Rasagiline in motor fluctuations]. PMID- 16217880 TI - From philosopher to psychologist: the early career of Edwin Ray Guthrie, Jr. AB - Edwin R. Guthrie rose to prominence as a psychologist in the 1930s. His theoretical outlook was behavioristic. This approach came from his conviction that an objective method could be applied to a scientific treatment of mind. Prior to becoming a psychologist, he was a philosopher of mathematics. Guthrie was initiated into psychology by Stevenson Smith, from whom he learned a psychology of adjustment informed by comparative research, Columbia functionalism, and clinical psychology. Guthrie's first step into psychology was in collaboration with Smith on Chapters in General Psychology (S. Smith & E.R. Guthrie, 1921). To synthesize their own unique position on learning from the contemporary theory and research, they used the principle of association. This articles focuses on Guthrie's origin and his development into a learning theorist. PMID- 16217882 TI - The mind of a rationalist: German reactions to psychoanalysis in the Weimar Republic and beyond. AB - In this article, the author seeks to trace the various attempts on the part of well-known German psychologists in the Weimar Republic to emphasize the rational side of psychoanalysis. In doing so, the author tries to demonstrate that the early reception in this period often resembled a critique of Freud's rationalism. It is possible to discern one particular form of criticism that emerged time and again, namely the association of psychoanalysis with the rationalist mind. If researchers wish to pinpoint further what lay beneath this purported connection, then it is possible to perceive a pronounced desire to prevent analysis of what many deemed to be sacred and beyond scientific scrutiny: the soul. It is precisely this discontent with Freud's thought that survived well into the Federal Republic, when other forms of critique had been discredited or no longer commanded serious attention. PMID- 16217883 TI - [Sports physiotherapy: a systematic overview]. PMID- 16217884 TI - The making of contemporary American psychiatry, Part 2: therapeutics and gender before and after World War II. AB - In this article, the 2nd in a 2-part series, the authors use patient records from California's Stockton State Hospital to explore the changing role of gender norms and other cultural values in the care of psychiatric patients. The authors show that cultural values are always imbedded in psychiatric practice and that their role in that practice depends on the patients, treatments, and therapeutic rationales present in a given therapeutic encounter. Because the decade following World War II witnessed dramatic changes in psychiatry's patients, therapeutics, and rationales, Stockton State Hospital's patient records from this time period allow the authors to show not only the extent to which gender norms shape psychiatric practice but also how psychiatry's expansion into the problems of everyday life has led to psychiatry taking a more subtle and yet more active role in enforcing societal norms. PMID- 16217886 TI - [Few remissions in aged patients]. PMID- 16217887 TI - [Mimic rehabilitation after removal of acoustic neurinomas: The Goettingen Concept]. PMID- 16217885 TI - Importance of the detection method for intact dimeric human chorionic gonadotropin without interference with the free human chorionic gonadotropin beta subunit for pregnancy exclusion before liver transplantation in a woman with cholangiocarcinoma. AB - Assay of human chorionic gonadotropin (hCG) is mainly used for the detection and monitoring of pregnancy, and for the follow-up of trophoblastic tumors. The serum free beta-hCG subunit (hCGbeta) is also a tumor marker in many non-trophoblastic tumors, including gastrointestinal cancers. In this work, we compared the performance of several immunoassays for pregnancy exclusion before liver transplantation and in the follow-up of a woman with cholangiocarcinoma. Serum hCG was detected with the Abbott Testpack plus hCG-Combo and measured with four automated sandwich immunoassays: ADVIA-Centaur, ACS:180, AxSYM and Dimension. hCGbeta was determined by an automated fluorescence sandwich immunoassay (Kryptor Free beta hCG) and with a specific immunoradiometric assay (ELSA-F beta hCG, Schering). The expression of hCG was also evaluated by immunohistochemistry on sections of intrahepatic cholangiocarcinoma cells and on peritoneal metastases. Before transplantation, discordant results were observed for pregnancy exclusion. Qualitative Testpack and Dimension tests detected no hCG-like immunoreactivity, whereas the ADVIA-Centaur, ACS:180 and AxSYM tests revealed positive levels. The same discrepancy was obtained in follow-up of the patient after liver transplantation. hCGbeta assay and immunohistochemical staining revealed tumor cell secretion of hCGbeta. In conclusion, a specific serum immunoassay for intact dimeric hCG without cross-reaction with hCGbeta should be adopted as routine policy for pregnancy exclusion before liver transplantation. PMID- 16217888 TI - [Surgical techniques. Nose, paranasal sinuses, midface, orbits, hypophysis VI]. PMID- 16217889 TI - Women in child development: themes from the SRCD Oral History Project. AB - Research in child development has emerged as a major intellectual discipline and a topic of great interest and importance to society. The Society for Research in Child Development (SRCD) has been the leading scholarly society concerned with the advancement of this research. As part of its focus on a history of the field, the SRCD has undertaken an oral history project in which prominent members are interviewed. Of the 102 completed interviews, 47 are women's accounts, which document important contributions women have made to child development. The experiences reported by individual participants reflect themes in the general history of women in psychology and child development during the early and mid 1900s. Women encountered obstacles to their efforts, even as they were supported by others and struggled to achieve prominence amid the sometimes ambiguous or conflicting circumstances that characterized women's entry into child development. PMID- 16217890 TI - Obituary. David Bakan (1921-2004). PMID- 16217892 TI - [Moyse Charas, an apothecary, a physician, as well as the author of "Pharmacopee royale galenique et chymique". The study of his dynasty]. AB - New research works conducted at the Archives nationales led to the discovery of numerous documents related to Moyse Charas's progeny and made it possible to complete former studies. Presently the filiation of this famous family of Parisians apothecaries of the seventeenth and eighteenth centuries is well established. A family tree has been drawn up. PMID- 16217891 TI - [Accidents in chemistry laboratories in the XIXth century]. AB - The author reviews the main accidents produced for most in French chemistry laboratories in the XIXth century: explosions, burns, serious poisonings. PMID- 16217893 TI - [The beginning of the teaching of chemistry in Lorraine: the Royal College of Medicine and the Faculty of Medicine of Nancy (1752 and 1776)]. AB - The faculty of medicine of the university created in Pont-a-Mousson in the second part of the XVIth century, transferred to Nancy in 1768, was not in possession of a chair of chemistry, but, in the middle of the XVIIIth century, it was interested by the development of this science. In Nancy, the Royal College of medicine, created by King Stanislas in 1752, disposed of a professor of chemistry since 1756, and also of a demonstrator who was one of the apothecaries of the town. In Metz, a course of this science occurred between 1756 and 1769 with the apothecary Thyrion. In 1776, the physician Henry Michel du Tennetar and the apothecary Pierre-Francois Nicolas opened a private course of chemistry, immediately transformed into a chair of the faculty of medicine. This chair will be maintained until Revolution. The personality and work of Michel du Tennetar and Nicolas, the circumstances and conditions of the creation of the chair, the inheritance of Michel, and the appointment of the demonstrator, also an apothecary, are successively described. PMID- 16217894 TI - [The Derosne, Parisian pharmacists from 1779 to 1855]. AB - The pharmacy of the 115 Saint Honore street was successively kept by Francois, by his two sons Jean-Francois and Louis-Charles, and by the son-in-law of the elder, Francois-Bernard who took the Bernard-Derosne family name. The two brothers made many chemical studies, among which the "essential salt" of the opium extract, and the sugar purification. Louis-Charles had many industrial activities, in Chaillot, where he made vapor engines. The two brothers were members of the Academie Royale de Medecine, and Jean-Francois was twice the President of the Societe de Pharmacie de Paris. PMID- 16217895 TI - [The separation of churches and chemist's shops. The "Tisane du cure de Deuil"]. AB - In 1945, in application of the law of September 11, 1941, the first-rate "Tisane du cure de Deuil" was almost renamed "Tisane de Deuil". Some documents found in the archives of Laboratories Lesourd allow to redraw, at least partially, this administrative imbroglio. PMID- 16217896 TI - [Short-term modeling of the effect of air pollution on health: analytical methods of time series data]. PMID- 16217897 TI - Suicide mortality in adolescents and young adults, 1980-1999. PMID- 16217898 TI - [The Book of the Eparch, by Leo VI: interest for the history of pharmacy]. AB - In 1893, Jules Nicole discovered a greek manuscript, named Genevensis 23, which proved to be the Book of the Eparch. This document contained the regulations for trade guilds in Constantinople, at the end of the IXth century and the beginning of the Xth century. It had been published by Emperor Leo VI, who reigned upon the Byzantine Empire from 886 to 911. Three professions mentioned in this book have some interest for history of pharmacy: wax chandlers, spicers and perfumers. The differences between spicers and perfumers consisted in the kind of products they sold and in the way they weighed these products. Spicers were, at the time, far from pharmacy, because they sold mainly food and no spices, while the perfumers dealt with spices and perfumes. Anyway, none of these professions was involved in preparing medicines. No mention was made of pigmentarioi in the Book of the Eparch and that raised the question of the part really played by these so-called "pigment makers" in the preparation and the dispensation of medicines. The role of physicians remained preeminent in supplying patients with drugs. PMID- 16217899 TI - [One hundred years after the Geminal law, Rene Cerbelaud publishes secret remedies]. AB - Our colleague Rene Cerbelaud was an original character. He played an important part in the pharmaceutical history of the 20th century when published recipes of countless medicines and cosmetic products, incurring the hostility of their manufacturers. As a matter of fact, he contributed to the disappearance of secret remedies and therefore to the enforcement of the Germinal law more than a century after it was promulgated. In a sense he may be looked on as the inventor of generic drugs and the promoter of what has become "parapharmacie". PMID- 16217900 TI - [The evolution of pharmaceutical schools in Toyama Prefecture since the Meiji era.]. AB - In this paper, the author attempts to generalize the evolution of pharmaceutical schools in Toyama since the Meiji Era. The copy of a petition drafted by pill peddlers of Toyama in 1873 suggests that there were expectations that a "Semi (chemicals)" school was going to be established; however, this was not realized. In 1877, the Ishikawa Prefectural Government established a small-scale pharmaceutical school associated with the public hospitals of Kanazawa, Toyama and Kukui. In 1883, Toyama Prefecture became isolated from Ishikawa Prefecture, and the above-mentioned pharmaceutical school continued operations until 1887, when the government closed it due to financial problems. Persuaded through a lecture given by Dr. Keizo Tampa, a professor of Tokyo University, volunteers in Toyama Prefecture established an independent pharmaceutical school in 1894. However, student enrollment was too low to maintain operation, and the administration started planning to relocate the school to the city of Toyama. This was realized in 1897. In 1900, a massive fire burnt down much of the city, severely damaging the school building and much of the equipment inside. Restoration would be costly. As a solution to the problem, the administration of the school was transferred from Toyama City to Toyama Prefecture. With money from the prefecture, the school facilities were repaired and teaching staff recruited. In 1909, the school ranking was promoted to college status, and it became one of the first academic institutions to issue a pharmacist licence to graduates without requiring them to take the national examination (i.e., with the exception of medical colleges). Subsequently, in 1920, the administration of the college was transferred from the Toyama Prefectural Government to Japan's Central Government, at which time the facilities were improved and a wider range of education subjects were introduced.The college was again relocated to the city suburbs, where the facilities included the building, a herbs garden and surrounding grounds. This successful move is known to be attributable to Dr. Nagayoshi Nagai, Professor of Tokyo University, as his enthusiasm aroused the interests of people in Toyama and other concerned authorities. The college was named Toyama Pharmaceutical Academy, and has since attracted many qualified applications from throughout the country. It has turned out prominent graduates for various occupations, and continues to remain well-known for its quality education. PMID- 16217901 TI - [Historical study on the introduction and cultivation of medicinal plants in the Edo-era. 1. A paper on the cultivation of 12 medicinal plant species.]. AB - This article describes a two-page response written by a translator in 1721, in which a reply is given regarding an inquiry made on October 5 to a Chinese person related to the cultivation of 12 medicinal plant species imported to Japan from China. One page, dated October 7, describes the cultivation method for seven species and the other page, dated October 10, describes the cultivation method for the other five species. These plants were imported to Japan for the purpose of domestic propagation as important materials for Chinese medicine at the Jyuzengi Medicinal Plant Garden in Nagasaki, and some of them were apparently sent to Koishikawa Medicinal Plant Garden in Edo (Tokyo). This is a historical document concerning when and which plants were imported for cultivation. PMID- 16217902 TI - [Use of crude drugs in "PiWeiLun" in comparison to those used in "NeiWaiShangBianHuoLun".]. AB - PiWeiLun (Hi-i-ron in Japanese) is a traditional Chinese medical formulary written in the 13th century that discusses endogenous disease. For curing endogenous diseases (Naisho) caused by functional deficiency of pi- and wei qi (Hi-I-Ki-Kyo), PiWeiLun was recommended for QingShuYiQi-Tang (Seisho-ekki-to), Ban-XiaBaiZhuTianMaTang (Hange-byakuju-tsu-tenma-to) and BuZhongYiQiTang (Ho-chu ekki-to), which are described in NeiWaiShangBianHuoLun (Naigaisho-benwa-ku-ron).A characteristic of PiWeiLun is that it discusses combining the use of Phellodendri Cortex with sweet Qi-tonics (Hoki-yaku) as Ginseng and Atractylodis Macrocephalae Rhizome for improving false heat syndrome (Kyo-netsu) due to yin-deficiency (In kyo). In PiWeiLun Cimicifugae Rhizome, Angelicae Radux and Atractylodes Rhizome are more frequently used than in NeiWaiShangBianHuoLun.The use of drugs presented in PiWeiLun may be useful to devise new crude drug formulations good for modern deficiency syndrome experienced by persons having post-operative complications. PMID- 16217903 TI - [Historical study of moth repellent, "Fujisawa Camphor" (6) - manufacturing and selling of "Fujisawa Camphor" during World War II.]. AB - During World War II, the amount of camphor production did not decrease, since it was used for munitions. At that time, camphor was not use for moth repellents, are not a life supporting necessity. The factory that took charge of camphor production was busy producing medicine for military use. Due to the war, an abnormal situation in the factory arose when the procurement department requested supplementation because of reinforcement of a lack of materials. Additionally, in the home, the use of moth repellent for clothing was not a concern. Of importance where was ensuring sufficient food to survive. The supply of "Fujisawa Camphor" for home use started in the post-war days, 1947. PMID- 16217905 TI - [The development of the Japanese pharmaceutical industry (Part 8) - the change of opium alkaloid opioid analgesics in Japanese pharmaceutical companies.]. AB - After the Japan-England Friendship and Commercial Treaty of 1858 (Ansei 5), narcotic drugs have been strictly regulated by laws. The production of opium alkaloid analgesics, mainly morphine, became active in Japan after World War I (1918, Taisho 7). Having released the "Cancer Pain Relief" guideline by the WHO in 1986 (Showa 61) and the morphine sulfate controlled-release tablet "MS Contin" in 1989 (Heisei 1) in Japan, the demand for morphine in Japan expanded remarkably. Today, several more kinds of strong opioid analgesics are available for cancer pain treatment in Japan. An increase in the use of strong opioid analgesics is expected in the future. PMID- 16217904 TI - [Pharmacological action of Gambir.]. AB - Gambir, which is called Asen-yaku in Japan, is mainly used as a stegnotic as well as an antiflatuent agent. The above data suggests, however, Gambir may also have anti-aging properties against the metabolic syndrome (MS) that has recently attracted concerns in the health community. While eating in moderation and appropriate exercise are naturally important in minimizing the MS, Gambir can play a role in improving each of the factors Prof. Kawamori presented as indicators of MS, including: 1. Improvement of visceral obesity, 2. Anti hyperlipidemic activity including reduction of triglycerides and cholesterols, 3. Normalization of blood glucose level, and 4. Normalization of blood pressure. PMID- 16217906 TI - [Historical study of the moth repellent, "Fujisawa Camphor" (7) - selling price and packaging.]. AB - The selling price of "Fujisawa Camphor" has increased 1,000-fold in the 100 years since it first went on sale. This 1,000-fold increase is not high at all. The price of "Fujisawa Camphor" was not low in the 1980s, one of grounds for setting the price was the damage caused by moths to expensive Japanese clothes and curios. For packaging in the Edo era, being wrapped in paper was the mainstream form, but modern-age packaging shifted to a cardboard box. As for "Fujisawa Camphor", a beautiful design was created for the cardboard. PMID- 16217907 TI - [Historical and pharmalogical study of Citrus hassaku.]. AB - The original plant of Citrus hassaku Hort. Tanaka was found at the Jyoudo Temple in Inno-shima, Hiroshima Prefecture, Japan by Ekei Shounin in the Edo Era. Citrus hassaku has been called "jagada" when it was first discovered. Since this fruit was seasoned for eating around "hassaku" (August 1st of the lunar calendar), it was given the scientific name as "C. hassaku." Today, the fresh raw fruits of C. Hassaku are cultivated as a seasonal food, and the most famous producing district of C. hassaku fruit is Wakayama Prefecture, representing about 60% of the Japanese output. The immature fruits of C. hassaku and its three main flavanone glycosides (naringin, neohesperidin and narirutin) was found to shown inhibitory activities on a compound 48/80 induced histamine released from mast cells, and anti-allergic effects on dinitrofluorobenzen (DNFB)-induced triphasic skin reactions in mice. The contents of the three flavanone glycosides in the immature fruits were higher than those in mature fruits. The application of extracts from the immature fruits of C. hassaku to skin care for patients with atopitic dermatitis has resulted in improving effects for 76% of the patients. Similar efficacy was obtained for patients pollinosis. PMID- 16217908 TI - [Fragments of information of Naoe Okamoto - the life of one of Japan's first female pharmacists.]. AB - It has generally been believed that Naoe Okamoto of Fukui Prefecture, who was licensed to open a pharmacy in December 1885, was Japan's first female pharmacist. However, there has been no way to confirm this because the pharmacists' registry held by the Ministry of Internal Affairs was destroyed by the Great Kanto Earthquake in 1923. This paper describes a hazy picture of her life, which we traced based on fragments of information from the Tokyo University Pharmacy and Life Science (TUPLS), pharmaceutical journals of her time, and two residents of Fukui City who met her when they were children. Naoe Okamoto was born in Asuwakamicho, Fukui City, in 1870, as the eldest daughter of a druggist operating since the Edo Era. Born in a socially and economically privileged family, Naoe went to Tokyo and entered Tokyo Pharmacists School (currently TUPLS) in 1883 or 1884. After two-years' study, she passed the pharmacist certification examination held by the Tokyo Prefecture Government in the autumn of 1885 and received a license to open a pharmacy from the Minister of Internal Affairs. In 1886 or 1887, she overtook the family business and married a doctor. She got divorced shortly thereafter, but continued to manage the family business although she had a younger brother. She died in 1941 at the age of 71. Her house and all articles left by her were burned to ashes after an air raid by the US Army Air Corps in July 1945 during the Second World War. Naoe left her small native city in Fukui Prefecture to study pharmacy in the capital at the beginning of the Meiji Era, when it was extremely difficult for women to pursue a professional career. She obtained the license to operate a pharmacy and pursued her career as a pharmacist through the Meiji, Taisho and Showa Eras. Her life offers great courage and lessons to today's 139,000 female pharmacists in Japan. Naoe Okamoto should be remembered as a pioneer of female pharmacists at this turning point when the 6-year pharmacy education system is about to start. PMID- 16217909 TI - [Rosa Gallica var. Officinalis.]. AB - I planted a "pharmacist's rose" in my garden about five years ago. It is a red rose of the Lancaster of "Red War" variety. The red rose that Tsunenaga Hasekura brought to "Entsu-ji" in Matshima is drawn on the door. Additionally, the crest used by the Imperial Household engraved by the balustrade. It turns out that it is the same as the one drawn on the ceiling as the "Rose War" monument. That is, the "pharmacist's rose" is a red rose of the Lancaster variety, and is expressed by the picture of an "Entsu-ji" rose. PMID- 16217910 TI - Handling Protocol of Posterior Composites Cavity Preparation. PMID- 16217911 TI - Financial literacy tools for young people with disabilities. PMID- 16217912 TI - Health care in Indian country. PMID- 16217913 TI - Hospital-acquired infection disclosure gains momentum. PMID- 16217914 TI - Texas passes first safe patient handling legislation. PMID- 16217915 TI - Courage and competency to collaborate. PMID- 16217916 TI - Nurses, fish and public health. PMID- 16217917 TI - Methylenetetrahydrofolate reductase (MTHFR) variants and bladder cancer: a population-based case-control study. AB - Functional variants in the methylenetetrahydrofolate reductase (MTHFR) gene, including the 677C>T and 1298A>C polymorphisms, have been associated with a moderately reduced risk of several cancers, including colorectal cancers. While recent studies have investigated the role of these polymorphisms on bladder cancer susceptibility, results have been mixed. To clarify the role of MTHFR polymorphisms on bladder cancer risk, we genotyped MTHFR 677C > T and MTHFR 1298A > C in a population-based study of bladder cancer of 352 patients and 551 controls from New Hampshire, USA. The allelic frequency was 35.6% for MTHFR 677C>T and 40.4% for MTHFR 1298A > C among controls. We found no evidence of a main gene effect for either polymorphism (adjusted OR for MTHFR 677C>T variants versus the reference genotype = 1.1; 95% CI, 0.8-1.4 and adjusted OR for MTHFR 1298A>C variants versus the reference genotype = 1.0; 95% CI, 0.7-1.4). Odds ratios did not appear to differ by smoking status or gender. We observed differences in the risk estimates for the MTHFR polymorphisms by arsenic exposure, but they were not statistically significant (P = 0.67 for MTHFR 677C > T and P = 0.12 for MTHFR 1298A>C). Thus, our findings do not support the presence of a main gene effect. The possibility that MTHFR polymorphism affects susceptibility to environmental exposures warrants further consideration. PMID- 16217919 TI - Is lead considered as a risk factor for high blood pressure during menopause period among Saudi women? AB - This case-control study was designed to examine the association between blood lead levels and high blood pressure in a restricted subpopulation, Saudi women who were 45-93-year old, during or after menopausal period and not occupationally exposed to lead. Blood lead levels were assessed in 100 women with hypertension and 85 control subjects. Lead concentrations were measured in the whole blood using flameless atomic absorption spectrophotometry. Blood pressure measurements were performed according to the World Health Organization recommendations. Results revealed that the mean blood lead levels for hypertensive were 47.52+/ 39.26 and 45.59+/-28.55 microg/l for controls. Participants were classified according to the median of blood lead levels in order to compute odds ratios. After controlling a number of potential confounding variables, the multiple logistic regression analysis revealed that women with blood lead levels of > or = 38.6 microg/l were 5.27 times more likely to be hypertensive than those with blood lead levels of < 38.6 microg/l, but of borderline significance (p = 0.06). Although such observation might support the hypothesis that the depletion of lead from bones during menopause increases blood lead levels placing women at increased risk for high blood pressure, there is a need for further studies with larger number of subjects. A number of risk factors, which were suspected to influence blood lead levels, were also investigated. Use of Kohl, duration of its use, osteoporosis disease and intake of calcium supplements were significantly associated with blood lead levels. PMID- 16217918 TI - Associations of pesticides, HCV, HBV, and hepatocellular carcinoma in Egypt. AB - The rate of hepatocellular carcinoma (HCC) is increasing in Egypt where the major risk factors are chronic infections with hepatitis B and C viruses (HBV and HCV). A major segment of the population is employed in agriculture, raising the possibility that exposure to pesticides is an additional risk factor for HCC. The objective of this study is to investigate pesticides as environmental risk factors for HCC while taking into account viral risk factors. We conducted a case control study of 236 subjects with confirmed HCC recruited from the National Cancer Institute, Cairo University, Egypt, and 236 controls matched on sex, age group and urban-rural status recruited from orthopedic department, Cairo University Hospital, Egypt. Patients who agreed to participate signed a consent form, answered a questionnaire and gave a blood sample for hepatitis virus testing. The manuals of the Ministry of Agriculture for approved use and type of pesticides since 1965 were linked to the questionnaire data for types of crops and pests that the subject had to combat, to attribute specific pesticides that were used by each subject. Subjects also reported duration of the exposure (years). Case-control comparisons in these data were stratified by sex, age group, and urban vs. rural residence. Data were analyzed using unconditional logistic regression models adjusting for age, HCV RNA, and current hepatitis B infection. Among rural males, the adjusted odds ratio (OR) for organophosphorus compounds was 2.7 (95% CI = 1.3-5.3) and for carbamates it was 2.9 (95% CI = 1.4 5.8). No statistically significant associations between HCC and pesticides were observed for urban males or for females. As expected, the strongest risk factors for HCC in this study were HCV RNA (OR = 16-17) and current HBV infection (OR = 27-28). This study therefore suggests that exposures to organophophorus and carbamate pesticides are additive risk factors to current HCV and HBV infection among rural males. Future investigation should address the possible hepatocarcinogenicity of pesticides using biomarkers of exposure and other techniques to better estimate dose-response relationships. PMID- 16217921 TI - Indoor environment and respiratory symptoms in children living in the Dutch German borderland. AB - OBJECTIVE: To investigate the relation between indoor environmental risk factors and respiratory symptoms in 7-8-year-old children living in the Dutch-German borderland. METHODS: A nested case-control study was conducted among children participating in a large longitudinal study on respiratory health. Parents of all 781 children with respiratory complaints and an equal number of randomly selected controls were asked to complete a questionnaire, including questions on indoor environment. RESULTS: The parents of 1191 children (76.2%) participated. Past exposure to environmental (OR = 2.73, 95% CI 1.14-6.67) as well as in utero exposure (OR = 2.28, 95% CI 1.15-4.53) to tobacco smoke, use of an unvented geyser for water heating (OR = 3.01, 95% CI 1.21-7.56), long-term exposure to dampness (OR = 2.98, 95% CI 1.10-8.28) or pets (OR = 2.18, 95% CI 1.39-3.42) increased the risk of asthmatic symptoms in 7-8-year-old children. A middle or low socio-economic status also increased the risk of asthmatic symptoms. An inverse association with asthmatic symptoms was seen for wall-to-wall carpeting (OR = 0.57, 95% CI 0.33-0.95) and insulation measures (OR = 0.46, 95% CI 0.25 0.83). Except for the presence of an unvented geyser, these environmental risk factors also presented a risk for coughing symptoms in children. CONCLUSION: This study showed an increased risk of respiratory symptoms in children exposed to several indoor environmental risk factors. PMID- 16217920 TI - Inhalational and dermal exposures during spray application of biocides. AB - Data on inhalational and potential dermal exposures during spray application of liquid biocidal products were generated. On the one hand, model experiments with different spraying devices using fluorescent tracers were carried out to investigate the influence of parameters relevant to the exposure (e.g. spraying equipment, nozzle size, direction of application). On the other hand, measurements were performed at selected workplaces (during disinfection operations in food and feed areas; pest control operations for private, public and veterinary hygiene; wood protection and antifouling applications) after application of biocidal products such as Empire 20, Responsar SC, Omexan-forte, Actellic, Perma-forte; Fendona SC, Pyrethrum mist; CBM 8, Aldekol Des 03, TAD CID, Basileum, Basilit. The measurements taken in the model rooms demonstrated dependence of the inhalation exposure on the type of spraying device used, in the following order: "spraying with low pressure" < "airless spraying" < "fogging" indicating that the particle diameter of the released spray droplets is the most important parameter. In addition inhalation exposure was lowest when the spraying direction was downward. Also for the potential dermal exposure, the spraying direction was of particular importance: overhead spraying caused the highest contamination of body surfaces. The data of inhalational and potential dermal exposures gained through workplace measurements showed considerable variation. During spraying procedures with low-pressure equipments, dose rates of active substances inhaled by the operators ranged from 7 to 230 microg active substance (a.s.)/h. An increase in inhaled dose rates (6-33 mg a.s./h) was observed after use of high application volumes/time unit during wood protection applications indoors. Spraying in the veterinary sector using medium-pressure sprayers led to inhaled dose rates between 2 and 24mga.s./h. The highest inhaled dose rates were measured during fogging (114 mg a.s./h) and after-high-pressure applications in the antifouling sector (110-300 mg a.s./h). The potential dermal exposure of spray operators was lowest (dose rates from 0.2 to 7 mg a.s./h) in the areas of food and feed disinfection and private and public hygiene during spraying with low-pressure devices. During fogging, wood protection and antifouling applications, high-potential dermal exposures of the operators were determined. Dermal dose rates varied between 100 and 34,000 mg a.s./h. PMID- 16217922 TI - Polymorphisms of DNA repair gene XPD and DNA damage of workers exposed to vinylchloride monomer. AB - Vinyl chloride monomer (VCM) is a human carcinogen. However, the exact mechanism of carcinogenesis remains unclear. 2-Chloroethylene oxide (CEO) and 2 chloroacetaldehyde (CAA), the metabolic intermediates of VCM, induce DNA damage which is mainly repaired by the nucleotide excision repair (NER) pathway. The XPD gene product and the related XPB protein are DNA helicases that are involved in transcription and NER. Polymorphisms of XPD have been implicated in chemical exposure-related health effects. In order to explore the mechanism of VCM-related health effects, a special matched case-control design (exposed workers with DNA damage and without damage) was used to investigate the association between the gene polymorphisms of XPD and DNA damage in 106 male and 44 female workers occupationally exposed to VCM. Exposure and anthropometrics information was collected through in-person interview. Such information was then used to calculate cumulative exposure doses of the workers. DNA damage in peripheral lymphocytes was measured by the single cell gel electrophoresis (SCGE) assay that identified DNA strand breaks. Genomic DNA from lymphocytes was used in genotyping assays. Genotypes of XPD Ile199Met, XPD Asp312Asn, and XPD Lys751Gln were identified by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) procedure. The results indicate that the genotypes of XPD 751Lys/Gln and Gln/Gln were significantly associated with the expression of DNA damages (OR 2.21, P<0.05, 95% CI 1.01-5.13). An interesting observation is the reduction of DNA damage for workers with high VCM exposure and possessing the XPD Asp/Asn and Asn/Asn genotypes (OR 0.33, 95% CI 0.11-0.95). Polymorphisms of XPD may therefore be a major reason of genetic susceptibility in VCM-induced DNA damage and health consequences. PMID- 16217923 TI - Transmissible spongiform encephalopathies: assessment of exposure risk in the histological working environment using GC-MS detection of fatty acids as marker for central nervous tissues. AB - To elucidate the risk of occupational exposure to the agent of transmissible spongiform encephalopathies (TSE) in the histological working environment, we assessed the principal suitability of three analytical methods for the detection of tissues of the central nervous system (CNT). We tested a neuron-specific enolase (NSE) Western blot, a glial fibrillary acidic protein (GFAP) ELISA and the GC-MS detection of some CNT typical fatty acids (FAs): omega9-tetracosenic acid, omega7-tetracosenic acid, lignoceric acid, and cerebronic acid. Histological sample processing (formalin fixation, dehydration, paraffin embedding) affected both of the immunochemical approaches considerably. The NSE Western blot produced negative results without exception. The results for the GFAP ELISA were better but still far too insensitive. Thus, both methods were judged to be unsuitable in their present form without major analytical adjustment. GC-MS sensitivity remained unaffected by the formalin fixation process. Sensitivity was reduced in the course of the final dehydration step using xylene in the histological sample processing. However, this reduction was found to be rather moderate (range 42-59%) when compared to the immunochemical methods. Overall, we judged GC-MS to be a promising analytical approach for the assessment of a potential TSE exposure risk via airborne CNT particles in the histological working environment. All the FAs we tested showed very low but detectable baseline contents. Thus, cut-off values must be used in the present GC MS approach. The most suitable FA turned out to be omega9-tetracosenic acid due to the greatest difference between its content in histological CNT samples and the respective cut-off value (689:1). Preliminary results by GC-MS monitoring of CNT via omega9-tetracosenic acid (and other FAs) on filters of routinely used vacuum cleaners and on filters after air sampling indicate that the airborne CNT/TSE exposure risk in the histological laboratory is minor if existing at all. However, further in depth studies will have to validate our preliminary findings and assess these results in the light of possible future data on human oral and/or pulmonary TSE susceptibility. PMID- 16217924 TI - Possible risk for re-colonization with methicillin-resistant Staphylococcus aureus (MRSA) by faecal transmission. AB - An increase in the frequency of methicillin-resistant Staphylococcus aureus (MRSA) as an important causative agent of nosocomial infections is observed worldwide. Unfortunately, in spite of optimal hygienic conditions (barrier isolation, screening, decontamination), patients often remain MRSA positive or are detected as "re-colonized" at readmission. The objective of our study was to clarify if this is due to an undetected colonization of the gastrointestinal tract, which could possibly lead to re-colonization after primary successful decontamination. Therefore, all MRSA strains were collected from 290 in-patients of a university hospital over a period of 2 years. A surprisingly high number (24.1% of all) was isolated from stool samples. Even 13.1% of the total collection could be first observed in this material before detecting MRSA in other materials of these patients. To evaluate the epidemiology of these isolates, pulsed-field gel electrophoresis (PFGE) was used. On the basis of PFGE restriction types one main clone and 11 singular clones could be identified. Additionally, for six individual patients MRSA isolates from stool specimens were indistinguishable from other isolates from different locations. We show here that colonization of the gastrointestinal tract with MRSA apparently could play an important role in spreading MRSA via faecal contamination. Hence, we suggest that stool colonization with MRSA could be the source of a so far unrecognized transmission of MRSA within individual patients and within the population. Therefore, our findings imply a modification in the hygienic strategies for handling decontamination and therapy of MRSA patients. PMID- 16217925 TI - In-line milk filter analysis: Escherichia coli O157 surveillance of milk production holdings. AB - Escherichia coli O157 is a major etiological agent of food-borne illness. Bovine animals are recognized reservoirs for this organism and represent a significant source from where these pathogens can enter the food chain. Food products derived from these animals are convenient vehicles, and are often the focal point(s) of infection. As a useful strategy to provide herd-level surveillance and to investigate for the presence of this pathogen in a population of Irish dairy cattle, milk filters from 97 farms were analysed by conventional culture and other methods. Five hundred and thirty-six milk filters were evaluated over a 2 year period. Filters from 12 of the 97 farms (12%) were found to contain E. coli O157, based on culture methods. Sixteen verocytotoxigenic E. coli O157 organisms were recovered and characterized in detail. The farm families in each case were consuming raw milk from their respective herds. The potential risk to public health associated with the detection of E. coli O157 and the local consumption of raw milk are discussed. PMID- 16217926 TI - Microbiological quality of carbonated drinking water produced with in-home carbonation systems. AB - The microbiological quality of carbonated water produced with tap water in commercial in-home carbonation systems was determined, the results being discussed in the context of the microbiological quality of the tap water used, the properties of the drink makers, and the procedures of preparation and washing of various parts of the appliance. The last-mentioned data were received from each participant of the study by questionnaire. Escherichia coli, coliforms, fecal streptococci and spore-forming sulphite-reducing anaerobes were used as indicators for the hygienic quality of the water. Tap-water samples were collected according to the usual procedure when filling the carbonating bottle, i.e., without previous flushing and disinfection of the faucet. In 12% of tap water samples, coliforms could be detected. On the other hand, in 20 of 52 carbonated waters (39%), coliforms as indicators of water pollution were found. By means of fecal streptococci and Pseudomonas aeruginosa, it was possible to establish additional contamination not involving E. coli or coliforms alone. Analysis revealed that, in addition to contaminated tap water, a bacterial biofilm on the inner surface of the re-usable bottles had a predominant influence on the microbiological quality of the carbonated water. PMID- 16217927 TI - Characterization of aeromonads isolated from the river Narmada, India. AB - Thirty isolates of Aeromonas species were isolated from river Narmada at Jabalpur during the period of January 2002-December 2002. Diversity of these fresh water isolates were determined by randomly amplified polymorphic DNA (RAPD) analysis. These environmental isolates were found to be positive for virulence factors, i.e. protease, amylase, lipase and DNase. Isolates were also positive for beta hemolytic activity. All Aeromonas species were tested for antibiotic resistance patterns and were found to be resistant to ampicillin and sensitive to gentamycin. PMID- 16217928 TI - Commentary regarding the article by Mutter et al. "Amalgam studies: disregarding basic principles of mercury toxicity" [int. J. Hyg. Environ. Health 207 (2004) 391-397]. PMID- 16217929 TI - Distributive justice in dentistry. PMID- 16217931 TI - Advice for a young editor: my journey in dental journalism. AB - Editing is sometimes a good route to writing; it may certainly be useful for an editor to understand the writer's point of view. Tips are offered regarding building writing skills, developing personal discipline, and generating story ideas. Writing and editing can be a way of finding out who one is. PMID- 16217932 TI - Advice for a young editor: you gotta love the details. AB - Writing and editing is a privilege of self-expression and an ethical responsibility to the reader. Know the process and take control of material, authors, schedules, printers, mailing, contacts and lists, and technology. Borrow where it makes sense and collaborate. PMID- 16217933 TI - Advice for a young editor: a long and satisfying relationship. AB - The mechanics and the details of dental journalism have changed as we have moved from the literal "paste-up" to the electronic world. The challenges of serving the interests of the readers and the sponsoring organization, of finding strong writers and writing the editorials have not. An emerging concern is ensuring that commercial interests do not exert unwarranted influence. There are resources- such as the American Association of Dental Editors, the American Dental Association, and editor colleagues--to help work through these issues. PMID- 16217934 TI - Advice for a young editor: a path to professional growth. AB - Journalism offers writers, editors, and the profession of dentistry an opportunity to share ideas. Editors must treat both writers and readers with respect. Legal standards must be understood and followed and commercial concerns are best addressed through disclosure practices. PMID- 16217936 TI - Advice for a young editor: the scientific publication. AB - Being the editor of a scientific journal involves the same challenges of working with authors, association officers, and readers as in all publications. In addition to the communication in the journal, editors must manage the communication about the journal. The editor's job involves building and maintaining trust. PMID- 16217935 TI - Advice for a young editor: an accidental career in dental journalism. AB - The editor of the Canadian Dental Association addresses issues such as balancing the interests of readers and the professional association, the value of mentors, building relationships with contributing authors and reviewers, and concerns over commercialism. PMID- 16217937 TI - Advice for a young editor: risks and rewards of dental journalism. AB - Editors and authors care to communicate with readers, and this concern guides their work. Politics is part of the job. There are organizations and various support for beginning editors, but becoming engaged with many colleagues and reading widely are essential for editors. Journalism is hard work. Constant innovation, rewriting, and resubmission are expected. PMID- 16217938 TI - Informed consent: does practice match conviction? AB - The need to obtain consent is an ethical principle and a legal requirement in health care that must be applied in practice. Dentists must give people the appropriate information about treatment options, risks, and benefits so that they are able to make informed choices before giving their free consent to any dental intervention. The British research reported on here supports the view that dentists generally have positive views about informed consent. But the companion program of observation of dentists in the clinic suggests that they do not have a systematic approach to obtaining consent and that patients are often told what is going to be done rather than asked. Dentists must approach informed consent more systematically, recognizing it as an integral part of treatment planning with the patient as developing the communication skills needed to do it well. PMID- 16217940 TI - Reflection. AB - Reflection is thinking about practice, based on traces of recalled facts or data, with a view toward better understanding it. Reflection is prompted by discrepancies between what we observe and what we expect and commonly takes the forms of comparing personal experiences, reviewing events mentally, looking for trends in data, building new prototypes, and managing impressions. Normally, the process involves awareness, reviewing selected traces, resolving tensions, and looking for the greater whole. Practitioners both reflect in practice (specific problem solving) and reflect on practice (drawing general conclusions). The essence of reflection is finding a meaningful way to make tacit knowledge available in explicit form. Often this involves story telling, with its attendant features of plot, emotion, dramatic tension, and the success of the hero. PMID- 16217941 TI - Cancer staging, prognostic factors, and our surgical challenges. PMID- 16217942 TI - Hemodynamic and pulmonary fluid status in the trauma patient: are we slipping? AB - Recent reports point to problems in the clinical assessment of the cardiopulmonary system in hemodynamically unstable patients, especially with the decreasing usage of pulmonary artery catheters. Our purpose was to evaluate the hypothesis that clinical judgment alone is inadequate for a reliable estimate of cardiopulmonary status in critically ill patients. Physician assessments (high, normal, or low) of cardiac index (CI) and thoracic fluid content (TFC) were made in 68 acute trauma cases and compared to the results obtained with impedance cardiography (ICG). Physician assessment using clinical judgment alone was correct only 42 per cent and 57 per cent, respectively, for CI and TFC. There was very little difference in heart rate (HR), blood pressure (BP), Glasgow Coma Score (GCS), and the number of injured systems between the incorrect and correct assessments of CI. However, the mean Injury Severity Score (ISS) was markedly higher for the incorrect than the correct CI values (18.8 +/- 9.3 vs 14.2 +/- 9.8, P = 0.0589). Thus, there is a need for an objective measurement of CI and TFC especially in the more severely injured patient. The inaccuracy of the clinical exam strongly suggests the need for a supplemental measurement, which the new and improved ICG monitor could provide. PMID- 16217943 TI - Prospective, blinded evaluation of accuracy of operative reports dictated by surgical residents. AB - Incomplete or inaccurate operative notes result in delayed, reduced, or denied reimbursement. Deficient reports may be more common when dictated by the surgical residents. We performed a blinded study to assess the accuracy of residents' dictations and their effect on the appropriate level of coding for reimbursement. A prospective, blinded study was performed comparing operative reports dictated by senior surgical residents (postgraduate years 3, 4, and 5) to reports dictated by attending surgeons. All residents had previously undergone group instruction on the importance and structure of operative notes. The trainees were blinded to the fact that the attending surgeons were dictating the operative reports on a separate dictation system. The dictations were analyzed by faculty reimbursement billing personnel for accuracy and completeness. Fifty operative reports of general surgical procedures dictated by both surgical residents and attending physicians were reviewed. A total of 97 CPT codes were used to report services rendered. Residents' dictations resulted in incorrect coding in 14 cases (28% error rate). The types of inaccuracies were a completely missed procedure (4) and insufficient documentation for an appropriate CPT code and/or modifier (10). All deficiencies occurred in complex, multicode, and/or laparoscopic cases. Sixty seven per cent of late dictations were incomplete. The financial analysis revealed that deficiencies in resident dictations would have reduced the reimbursement by $18,200 (9.7%). For cases with deficient dictations, 29.5 per cent of charges would have been missed, delayed, or denied if the resident dictated note was used to justify charges. Operative reports dictated by surgical residents are often incomplete or inaccurate, likely leading to reduced or delayed reimbursement. Dictations of complex, multicode, or laparoscopic surgeries, especially if delayed beyond 24 hours, are likely to contain significant deficiencies that affect billing. Attending surgeons may be better equipped to dictate complex cases. Formal housestaff education, mentorship by the attending faculty, and ongoing quality control may be paramount to minimize documentation errors to ensure appropriate coding for the services rendered. PMID- 16217944 TI - The general surgery model: a more appealing and sustainable alternative for the care of trauma patients. AB - The contemporary model of trauma care where dedicated trauma/critical care surgeons exclusively manage trauma patients has become progressively unsustainable. Little objective data, however, is available documenting that a better model exists. From September 2002 through August 2003, the trauma model at a 735-bed level I trauma teaching hospital was changed from the contemporary model to a new one where selected general surgeons with Advanced Trauma Life Support (ATLS) certification covered in-house trauma and emergency surgery call on a rotational basis. As well, each pursued elective practices, admitting all inpatients (trauma, emergent, elective) to a single teaching service (formerly the trauma service). Critical care was managed by a separate group of intensivists. The purpose of this study was to objectively compare the two models. Quantitative, financial, and qualitative data were derived from August 2001 to January 2002 (trauma/critical care model) and compared to August 2003 to January 2004 (general surgery model). During the two periods (trauma/critical care vs general surgery), the mean Revised Trauma Score (7.1 vs 7.2; P = 0.029), the mean Injury Severity Score (ISS) (10.9 vs 10.8; P = 0.84), and the percentage of penetrating trauma (12.5% vs 13.2%; P = 0.79) were similar. Differences (trauma/critical care vs general surgery, % increase/P value) included average daily census (24 vs 54, 225%), cases/attending (262 vs 543, 207%), cases/resident (54 vs 262, 485%), charges/attending (353,811 dollars vs 471,725 dollars, 133%), collections/attending (106,143 dollars vs 165,103 dollars, 156%), number of trauma patients (643 vs 748, 116%), trauma mortality (7.3% vs 4.0%; P = 0.007), trauma mortality with ISS >15 (21.7% vs 12.0%; P = 0.035), trauma complications (33.1% vs 17%; P < 0.001), and ICU morbidity (66.8% vs 43.9%; P < .001). The new general surgery model produced superior financial results and better quantitative surgical experience while exceeding trauma and ICU quality outcomes compared to the former trauma/critical care model. These data objectively support a model such as ours--one that is financially sustainable and more professionally attractive. PMID- 16217945 TI - Postoperative outcomes according to preoperative medical and functional status after infrainguinal revascularization for critical limb ischemia in patients 80 years and older. AB - The purpose of this study was to provide outcomes after intervention for critical limb ischemia (CLI) in elderly patients (> or =80 years) according to medical and functional status at presentation. From January 1998 to September 2003, 140 limbs/122 patients (age range 80-97 years) were treated (57 patients/66 limbs, infrainguinal bypass; 65 patients/74 limbs, infrainguinal angioplasty) for CLI. At presentation, 71 (58.2%) patients were functionally ambulatory, 41 (33.6%) were homebound ambulators, and 10 (8.2%) were transfer-only ambulators. Overall end points after treatment as well as outcomes according to type of treatment and preoperative medical and functional status were determined. End points included reconstruction patency, limb salvage, survival, amputation-free survival, and maintenance of ambulatory and independent living status. Results for the 140 limbs/122 patients at 3 years (Kaplan-Meier curves) include primary patency, 55.3%; secondary patency, 73.2%; limb salvage, 78.3%; survival, 62.5%; amputation free survival, 49.7%; maintenance of ambulation, 77.8%; and maintenance of independent living status, 82.9%. There was essentially no difference in outcomes based on type of treatment (endovascular vs open operation). When analyzing 2 year outcomes by functional status (ambulatory vs homebound vs transfer), there was deterioration in outcomes according to declining functional status at presentation for mortality (84.7% vs 66.4% vs 42%; P < 0.001), amputation-free survival (73.3% vs 48.2% vs 36.9%; P < 0.001), limb salvage (86% vs 66.5% vs 71.9%; P = 0.022), and secondary patency (84.3% vs 61.5% vs 69.2%; P = 0.005) regardless of treatment. Homebound ambulators were two times and transfer-only patients five times more likely to experience death (Cox hazard model); diabetics were four times more likely to lose a limb and experience a decline in ambulation and living status. Overall medical and functional status at presentation predicts postoperative functional outcomes. These data support a policy of aggressive vascular intervention in the functional elderly and clinical restraint in the functionally impaired patient with CLI. PMID- 16217946 TI - Carotid endarterectomy: update on the gold standard treatment for carotid stenosis. AB - Many prospective, randomized clinical trials evaluating the safety and efficacy of carotid endarterectomy (CEA) versus medical management in the prevention of ischemic stroke were performed in the 1990s. Clinical trials are underway that will compare CEA outcomes to carotid stenting; however, relatively few studies have examined the outcomes of modern CEA. The purpose of this report is to examine current outcomes of CEA and evaluate hospital costs and length of stay. Statewide results were collected for all hospitals, except Veterans Administration hospitals, by Virginia Health Information (VHI). Data for the years 1997-2001 were evaluated, and data were based on the All Patient Refined Diagnostic Related Group (APR-DRG; 3M Company). A total of 14,095 CEAs were performed in a 5-year period. The mortality of patients undergoing CEA was 0.5 per cent. The stroke rate was 1 per cent overall and decreased each year of the study. Mean and median lengths of hospital stay were 3 and 2 days, respectively. Length of stay decreased over the course of this study. Mean and median hospital costs were 14,331 dollars and 11,268 dollars. Higher rates of mortality and stroke and higher costs were observed at low-volume hospitals. The need for CEA is substantial. CEA is safe and inexpensive. The data presented here demonstrates continued refinement in CEA, leading to a very low rate of perioperative adverse events, declining lengths of stay, and low hospital costs. PMID- 16217947 TI - Emergent right hemicolectomies. AB - Emergent right hemicolectomies have historically been associated with surprisingly high morbidity and mortality rates. A retrospective review of emergent right hemicolectomies over a 7-year period was performed to assess current morbidity and mortality. Emergent right hemicolectomy was defined as a procedure performed for an acute abdomen with no formal preoperative cleansing of the colon. Demographic data, diagnostic evaluation, length of stay and outcomes were evaluated. Over the study period, 122 emergent right hemicolectomies were performed on both general surgery and trauma patients. The average patient was 52.9 +/- 18.5 years old, and the majority of patients (66.4%) were male. The indications for the procedures performed were bowel perforation (51), hemorrhage (25), cancer (16), benign obstruction (14), phlegmon (8), ischemia (6), or other (2). Resection with primary anastomosis was performed in 98 patients, 16 had an end ileostomy, and 8 underwent damage control procedures in which gastrointestinal continuity was not reestablished at the time of the original operation. Postoperative complications developed in 48 patients (39.3%). The majority of the complications (83.3%) were related to infection including intra abdominal abscess (21 patients), sepsis (16), and wound infection (5). Other complications included anastomotic leak (5), wound dehiscence (3), stoma-related (3) and postoperative bowel obstruction (2). The patients who developed complications did not differ from those who had an uneventful postoperative course in terms of age, indication for procedure, or presence of intraabdominal abscess or gross contamination at the time of the original procedure. The overall mortality rate was 13 per cent. Patients who died were older than those who lived (63 +/- 19 vs 52 +/- 18; P = 0.03) and were significantly more likely to have evidence of shock on presentation (P = 0.0013). Emergent right hemicolectomies continue to be associated with high morbidity and mortality rates. The most common complications are related to infection. Age and manifestations of shock at the time of admission are strong predictors of mortality. PMID- 16217948 TI - Absorbable cyanoacrylate as a vascular hemostatic sealant: a preliminary trial. AB - No absorbable cyanoacrylate tissue adhesive for safe internal surgical use is available. This prospective multicenter preliminary study was designed to evaluate the safety and effectiveness of an investigational absorbable cyanoacrylate (IAC) as an adjunct to hemostasis in arteriovenous shunt (AVS) procedures for dialysis access. Consenting adults (10) underwent placement of expanded polytetrafluoroethylene (ePTFE) upper extremity vascular grafts for AVS access using continuous 5-0 or 6-0 polypropylene after heparinization (> or =3000 units i.v.). Arterial anastomoses were evaluated after sealing with IAC followed by 120 seconds of polymerization time. After vascular clamp removal, the mean time to hemostasis was 9.1 +/- 28.8 seconds. Additionally, 90 per cent (9/10) and 100 per cent (10/10) achieved hemostasis by 1 and 5 minutes, respectively. No patients required further adjunctive hemostatic measures. Adverse event safety data analysis through 12 weeks revealed occlusion of graft or vessel in four patients and graft thrombosis in one patient, all thought unrelated to sealant use. Other unrelated adverse events (bleeding, death, deep venous thrombosis, edema, erythema, hematoma, infection, and rash) occurred in single patients. Thus, IAC could be a useful sealant for vascular procedures with a potentially satisfactory safety profile. Larger, randomized, multicenter, prospective trials to further evaluate the use of this material are indicated and appropriate. PMID- 16217949 TI - A preliminary study of imiquimod treatment in variants of basal cell carcinoma. AB - Imiquimod is a topical immune response modifier that has proved efficacious in the treatment of the superficial variant of basal cell carcinoma. The nodular variant of basal cell carcinoma has shown moderate response to imiquimod; other variants have not been tested. The mechanism of action is largely unknown; however, studies indicate the mechanism involves alteration of local cytokine production. The objective of this study is to evaluate the cytokine response of imiquimod in all variants of basal cell carcinoma. Ten patients were selected who had clinically and histologically proven basal cell carcinoma. All lesions were treated with imiquimod once a day, 5 days a week, for 3 weeks. After a 3-week rest period, the lesions were rebiopsied. All biopsy specimens were analyzed via polymerase chain reaction (PCR) for various cytokines. Nine of 10 lesions resolved clinically, which included nodular, superficial, infiltrative, adenoid, and micronodular variants. The cytokine with the greatest change pre- and post treatment was IL-8, which decreased an average of 44 per cent (P = 0.06). We concluded that topical 5 per cent imiquimod is an effective treatment of various subtypes of basal cell carcinoma. IL-8, which plays an important role in the development and metastasis of melanoma, may be involved in the mechanism of action of imiquimod on cutaneous malignancies. Larger studies are needed to prove the efficacy of imiquimod on nonsuperficial variants of basal cell carcinoma and cutaneous melanoma metastasis. PMID- 16217950 TI - Tailgut cyst (retrorectal cystic hamartoma): report of a case and review of the literature. AB - Tailgut cysts are rare congenital lesions arising from remnants of normally regressing postanal primitive gut. They often present in middle-aged women with perirectal symptoms and a retrorectal multicystic mass. These cysts have occasionally shown malignant transformation. We report a case of a tailgut cyst occurring in a 25-year-old African-American female. The differential diagnosis of a retrorectal mass is briefly explored, and the etiology, diagnostic strategy, and surgical approach for tailgut cysts is examined. We also report an extensive literature review to examine clinical characteristics and surgical data for 43 cases of tailgut cysts spanning 16 years. PMID- 16217951 TI - A survey of the work effort of full-time surgeons of the Southeastern Surgical Congress. AB - This is a survey research project to determine the work hours of practicing surgeons and compare those hours with hours that have been mandated for graduate medical education programs by the Accreditation Council for Graduate Medical Education (ACGME). The survey conducted of the membership of the Southeastern Surgical Congress focused on the amount of time devoted to professional activity. Although several categories of membership were surveyed, those surgeons in full time practice were used for this report. Five hundred ninety-two general surgeons and some surgical specialties from 17 states reported a total professional work effort of 65 hours per week averaged over a month. Twenty per cent reported working more than 80 hours per week. Statistically significant (P < 0.05) factors that characterized these individuals included years in practice (1 to 10 years), more clinical hours per week, fewer administrative hours per week, fewer teaching hours per week, fewer continuing medical education (CME) hours per year, and an increase in recent clinical practice. Interestingly, there was no significant difference in CME over a 2- or 5-year period. Other factors such as type of practice did not have statistical significance. There was no difference between states and no difference in time commitment to political or community activities. This survey indicates that surgeons going into practice in the Southeast from general surgery graduate medical education programs can expect to have a mean work week of 65 hours, and 20 per cent can expect to exceed an 80-hour work week. PMID- 16217952 TI - Early versus delayed single-stage laparoscopic eradication for both gallstones and common bile duct stones in mild acute biliary pancreatitis. AB - Several studies addressed that preoperative endoscopic retrograde cholangiopancreatography (ERCP) for common bile duct (CBD) clearance, followed by interval laparoscopic cholecystectomy (two-stage approach), constitutes the most common practice in cases of uncomplicated mild acute biliary pancreatitis. Between June 1998 and December 2002, 44 patients (35 females and 9 males with a median age of 62 years) suffering from uncomplicated mild acute biliary pancreatitis were treated in our unit. All patients were electively submitted to surgery after subsidence of the acute symptoms, and for definitive treatment we favored the single-stage laparoscopic management, avoiding preoperative ERCP. All patients underwent laparoscopic cholecystectomy plus fluoroscopic intraoperative cholangiogram (IOC). If filling defect(s) were detected in the IOC, a finding suggestive of concomitant choledocholithiasis, laparoscopic common bile duct exploration (LCBDE) was added in the same sitting. Twenty patients were operated upon within 2 weeks since the attack of the acute symptoms and constitute the early group (n=20), whereas 24 patients underwent an operation later on and constitute the delay group (n=24). We retrospectively compare the safety, effectiveness, and outcome after the single-stage laparoscopic management between the two groups of patients. Laparoscopic cholecystectomy alone constituted the definitive treatment in 38 patients, while an additional LCBDE was performed in the remaining 6 patients (14%), and all operations were achieved laparoscopically. There was no statistically significant difference between the groups in terms of operative time, incidence of concomitant choledocholithiasis, morbidity rate, and postoperative hospital stay. During the follow-up, none of the patients experienced recurrent pancreatitis. In uncomplicated mild acute biliary pancreatitis cases, a single-stage definitive laparoscopic management, avoiding preoperative ERCP, can be safely performed during the same admission, after the improvement of symptoms and local inflammation. Postoperative ERCP should be selectively used in patients in whom the single-stage method failed to resolve the problem. PMID- 16217953 TI - Roux-en-Y diversion for debilitating reflux after esophagectomy. AB - Two patients with debilitating reflux after esophagectomy are reported. Complete relief of symptoms after creation of a Roux-en-Y limb to the gastric conduit is described. PMID- 16217954 TI - A needs assessment for regionalization of trauma care in a rural state. AB - Systems of trauma care in urban areas have a demonstrated survival benefit. Little is known of the benefit of trauma system organization in rural areas. We hypothesized that examination of all trauma deaths during a 1-year period would provide opportunities to improve care in our rural state. We used a medical examiner database of trauma deaths occurring during a 1-year period. Five board certified surgeons analyzed deaths as preventable (P), potentially preventable (PP), and non-preventable (NP) using modified Delphi technique. There were 223 trauma deaths during a 1-year period. Most (63%) died at the scene prior to any medical intervention. Adjudication of the deaths demonstrated 5 P (2%; 95% CI 1 5%), 36 PP (16%; 95% CI 12-27%), and 179 NP (81%; 95% CI 76-86%). Agreement among trauma surgeons was only moderate with a k of 0.46. Suicide accounted for a significant number of the overall trauma deaths at 32 per cent. Rural trauma system design should focus on discovery, as that is where the majority of deaths occur. Suicide is a significant problem in this rural state that should be aggressively targeted with prevention programs. PMID- 16217955 TI - An unusual rectosigmoid mass: endometrioid adenocarcinoma arising in colonic endometriosis: case report and literature review. AB - Malignant transformation is an infrequent complication of endometriosis. The ovary is the primary site in 79 per cent of cases, and extragonadal sites are identified in 21 per cent. Primary involvement of these types of tumors with the colon and/or rectum is a rare clinical entity. Endometrioid carcinoma is a common histologic type that remains a diagnostic challenge-the main differential diagnosis includes colorectal carcinomas. We report a case of malignant transformation arising in colonic endometriosis. The patient had a total abdominal hysterectomy and bilateral salpingo-oophorectomy 10 years before she presented with hematochezia. The patient was ultimately treated by surgical resection. Immunohistochemical staining in addition to the usual histopathology was critical for accurate diagnosis of this endometriosis-associated intestinal tumor. PMID- 16217956 TI - Minimally invasive management of obstructive gastroduodenal tuberculosis. AB - Pulmonary tuberculosis (TB) is prevalent in Western urban centers, especially among immunocompromised patients and immigrants. However, TB enteritis is a rare sequela, occurring in less than 1 per cent of this population. Tuberculosis may affect any portion of the gastrointestinal (GI) tract, and 85 per cent of cases manifest in the ileocecal region. However, the stomach and duodenum are involved in just 0.3-2.3% of TB cases that affect the gut. Gastric outlet obstruction due to TB has been traditionally treated by a surgical bypass operation, followed by anti-TB chemotherapy. In a recent review of 17 cases of TB-related gastric outlet obstruction, gastrojejunostomy or duodenojejunostomy was performed in all patients. We present a case of gastric outlet obstruction due to TB that was treated successfully with a minimally invasive approach, without the need for a gastric bypass. PMID- 16217958 TI - [A possible mechanism of influence of modifiable striatal lateral inhibition on the conditioned selection of motor activity]. AB - A mechanism of the influence of dopamine-evoked modulation of lateral inhibition in the striatum on a conditioned selection of motor activity is proposed. According to suggested modulation rules for inhibitory transmission, action of dopamine on postsynaptic D1 (D2) receptors on striatonigral (striatopallidal) cells promotes long-term depression (potentiation) of inhibitory inputs simultaneously with potentiation (depression) of "strong" excitatory inputs that open NMDA channels on these neurons. If excitatory inputs are "weak" and NMDA channels are closed, modulation rules have opposite signs. Activation of presynaptic D2 (D1) receptors results in a decrease (increase) in GABA release from striatopallidal (striatonigral) axon terminals that innervate striatonigral (striatopallidal) cells. Thereof, dopamine-evoked modulation of lateral inhibition simultaneously strengthens both potentiation (depression) of excitatory inputs to "strongly" activated striatonigral (striatopallidal) neurons rising (reducing) their activity, and depression (potentiation) of excitatory inputs to "weakly" activated striatonigral (striatopallidal) neurons reducing (rising) their activity. Subsequent reorganization of neuronal activity in the cortico-basal-ganglia-thalamocortical loop promotes a conditioned selection of motor reaction because of the further increase (decrease) in activity of those motocortical neurons that "strongly" ("weakly") activated the striatum during dopamine release in response to conditioned stimulus. PMID- 16217957 TI - [Suppression of gamma EEG activity as an index of a spreading depression wave in the neocortex of a waking rabbit]. AB - A spreading depression (SD) can spontaneously develop in seizures, attacks of migraine, vascular disorders and other pathological states of the brain. However, problems in technique of recording the DC-potential in the neocortex of humans and waking animals substantially restrict the possibilities of studying functional consequences of the SD. In this article, the EEG pattern was studied in detail at the moment of the SD development. Specific features were revealed, which make it possible to detect the SD without recording shifts of the DC potential. At the moment of the SD arrival, the interhemispheric balance drastically disturbs because of a strong decrease in the high-frequency activity. By the time indices, the course of the suppression of the gammal and gamma2 EEG frequencies is the most reliable symptom of the SD wave development. The EEG spectral power in the delta band increases with a certain delay in reference to the deep depression of the high-frequency activity and is, in essence, an SD aftereffect. The found EEG signs of an SD wave can substantially simplify the identification of this phenomenon both in experiment and clinical conditions in certain pathological states of the brain. PMID- 16217959 TI - [Neurodarvinism: models of selection of neuronal groups]. AB - Concept of neurodarvinism is regarded in the context of simulation of the "natural" and "artificial" selection of neurons, synapses and neuronal groups. "Natural" selection of neurons is based on mobile devices built of neuron-like elements. These devices should be capable for adaptation to real surrounding. "Artificial" selection of neurons is performed using computerized "neurointelligence" model operating in a virtual environment. Comparison of the models suggests the advantage of the integration of these approaches. PMID- 16217960 TI - [Effects of spatial-imagery task performance on the heart rate variability]. AB - Effects of the efficiency of spatial task performance on the autonomic heart control were studied. Dynamic changes in autonomic balance diagnosed on the heart rate variability during maze-model task performance (the type of spatial-imagery activity) showed that the type of heart rate regulation depended on the performance efficiency. The results suggest that in subjects with low performance efficiency, problems in performance are associated with negative emotions that produce the emotional psychogenic stress. In subjects with high efficiency, the autonomic contour of the heart rate regulation prevailed. This was presumable associated with the adequacy of the maze-model task. PMID- 16217961 TI - [Classification of the adolescent EEG by the spectral and segmental characteristics for normals]. AB - The 16-channel EEG records of 45 adolescents with schizophrenia and 39 healthy adolescents were subjected to statistical combinatorial analysis of 160 elementary EEG characteristics (6 spectral and 4 segmental EEG characteristics for a channel). Employing pattern recognition algorithm "Kora-n", a list of 37 combined EEG patterns was compiled. This list characterized with a minimal error the EEG of healthy adolescents in such a way that none of these characters featured the EEG of adolescents with schizophrenia. Analysis of this list of EEG characteristics suggests that the contrast between EEG of healthy and ill adolescents is the sharpest in the F4, Cz, T3 and O1 derivations. Compared to EEG samples of schizophrenic subjects, EEGs of healthy subjects exhibit lower levels of delta and theta activity mainly in the frontal and temporal regions of the cortex and higher level of alpha activity predominantly in the occipital region. Applicability of the list of EEG patterns for diagnostics of schizophrenia-type disorders of adolescents is discussed. PMID- 16217962 TI - [Frequency-spatial organization of brain electrical activity in creative verbal thinking: role of the gender factor]. AB - Gender differences in EEG patterns associated with verbal creativity were studied by EEG mapping. The EEGs of 18 males and 21 females (right-handed university students) were recorded during a performance of Remote Associates Task (RAT) compared with the letter-fluency and simple associate's tasks. Gender differences were found in a factor structure of the indices of verbal thinking and a score of generating words was greater in women than men. No significant gender differences in originality of associations were revealed, however, gender-related differences in the EEG-patterns were found at the final and initial stages of RAT. In men, the beta2-power was increased in both hemispheres at the beginning of test. To the end of testing, the power of oscillations in the beta2 band increased only in the central part of the cortex. In women, the beta2-power was increased to a greater extent in the right than in the left hemisphere at the initial stage of task performance, whereas the final stage was characterized by a relative decrease in beta-activity in parietotemporal cortical regions and increase in the left prefrontal region. It is suggested that the verbal creative thinking in men is based mostly on "insight" strategy whereas women additionally involve the "intellectual" strategy. PMID- 16217963 TI - [Correspondence between spectral power and synchronization of the brain rhythms in the norm and cognitive pathology]. AB - The work is aimed at the study of correlations between the measures of spectral power and cortical interactions of EEG rhythms in healthy subjects and schizophrenic patients ("acute" and chronic cases). All brain rhythms in healthy subjects appeared to be symmetrical and synchronous both in phase and frequency. In "acute" schizophrenics, opposite to healthy subjects, the distribution of cortical activity is asymmetrical, and in the chronic cases, the spectral power of most cortical rhythms is decreased as compared to healthy subjects. In the "acute" patients, interhemispheric connections are absent in all rhythms but alpha. In the chronic patients, the number of cortical connections is slightly higher than in the acute patients; and they are located in the posterior areas in the gamma rhythm. These neurophysiological aberrations evidently underlie the multiple mental activity disorders in schizophrenic patients. Thus, the correspondence between the brain rhythms and their synchronization is a necessary condition for normal perception, emotions and cognition evidently influencing behavior and consciousness. PMID- 16217964 TI - [Positive potentials ot the human brain in the latent presaccadic period under conditions of stimulation of the leading and nonleading eye]. AB - We used backward averaging method to study fast positive presaccadic EEG potentials under conditions of the monocular stimulation of the leading and nonleading eye. Two schemes of the visual stimulus presentation ("no gap" and "overlap") were used. In the "no gap" condition, potential P1 dominated in the hemispere ipsilateral to a saccade direction. In the "overlap" condition, when the gaze was fixed at the central point, foci of this potential were localized in the sagittal derivations or in the same sites as in the "no gap" conditions. Irrespective on the stimulation scheme, the P2 foci were localized in the hemisphere contralateral to a saccade direction. We assume that the fast positive potentials involve both activation and inhibition processes in visuomotor structures and can be also associated with cognitive presaccadic processes (such as fixation disengage, attention lateralization and a preliminary extraction of motor programs from memory). PMID- 16217965 TI - [Habituation of evoked potentials in subjects with high, medium and low levels of anxiety]. AB - The habituation of the peak-to-peak amplitudes of N1P2 and P2N2 complexes of the evoked potentials as responses to series of the increasing number of visual and acoustic stimuli was investigated in 55 male and female volunteers aged 20-30. The size and speed of the habituation process were approximated by hyperbola. The shape and coefficients of the hyperbola were correlated with high, medium and low levels of the anxiety and probably reflected the balance between synchronization and desynchronization processes in the cortex. PMID- 16217966 TI - [Emotionality and spatio-temporal organization of momentary EEG potentials]. AB - With the aim to reveal features of the neocortical spatiotemporal organization of potentials characteristic of different genetically predetermined emotional levels, momentary values of EEG potentials were analyzed in inbred rats of MR and MNRA strains. A topogram was described by a basic parameter such as its general level (the mean of momentary values of potentials derived from 24 symmetrical electrodes bilaterally implanted into the brain cortex) and a similarity coefficient (correlation between the set of its momentary values and that of a standard topogram). The general level and similarity coefficient values were calculated for a series of successive topograms individually for the right and left hemisphere. Also, right- and left-side power spectra of these series were calculated. In rats of MR strain, significant (p < 0.05) peaks in the general level and similarity coefficient spectra were observed in the delta (2.0 Hz), teta (6.5 Hz) and alpha (9 Hz) frequency bands. In this strain, the general level power was higher at the right side, and the similarity coefficient power displayed the left-side dominance. In rats of MNRA strain, peaks in the delta(2.0 Hz) band coinsided in the general level and similarity coefficient spectra, whereas, independently, the general level spectra had peaks in the theta band (7.0 Hz), and similarity coefficient had peaks at frequencies 3.0, 4.5, and 6.0 Hz. The left-side general level spectral power was higher than the right-side general level spectral power in the delta and lower in the high-frequency theta bands. The similarity coefficient power displayed the left-side dominance for the peaks in the delta (2.0 Hz) and theta (3.0 Hz) bands, and it displayed the right side dominance for the peaks in the theta (4.5 Hz) band. The specific features of the cortical spatiotemporal organization of potentials revealed in rats of MR and MNRA strains suggest different modes of functioning of at least two systems, reticulo-thalamo-corticaland hippocampo-cortical. PMID- 16217967 TI - [Hippocampal neurons of the CA3 field in animals of different typological groups exposed to emotional stimuli]. AB - Hippocampal unit activity in of the right and left CA3 fields was studied in rats which were divided in two groups by the method of "emotional resonance": the animals that did ("A") and did not stop ("E") crying of rat-partner. The rate of neuron firing was studied in the state of hunger, satiation, and under exposure to intracranial electrical stimulation of the emotional positive and negative structures of the brain. It was shown that the intracranial stimulation, especially positive, increased the rate of firing. Under exposure to emotional stimuli, the neural activity was asymmetric only in E rats. The activity at the left was more intensive then the right-side activity and did not depend on the sigh of emotion. PMID- 16217968 TI - [Dual manifestation of catatonic reaction in rats]. AB - Results of genetic, neurophysiological, neurochemical and pharmacological suggest that the cataleptic freezing and "nervousness" observed in the cataleptic rat strain GC have a common mechanism. There seems to be a physiological factor causing catalepsy, upon reaching a certain level of intensity, to be transformed into "nervousness", which is observed both at different period and/or moment of individual life and in the processes of breeding the strain for predisposition to catalepsy. PMID- 16217969 TI - [Influence of D1 dopamine receptor activation on extinction of passive avoidance and amnesia in aggressive and submissive mice]. AB - In mice with aggressive and submissive behavioral stereotypes, passive avoidance retrieval in extinction and amnesia was shown to be differently dependent on the activation of D1 dopamine receptors. In extinction, agonist of D1 receptors SKF 38393 injected before training or on 12-th day of testing in a dose of 5 mg/kg impaired the retrieval of a conditioned habit in aggressive mice and improved the retrieval in submissive mice. The opposite effects of D1 receptor activation depending on stereotype were also observed in a procedure of amnesia. In aggressive mice, SKF 38393 essentially reduced the resistance to amnesic effects characteristic of this stereotype. In submissive mice, SKF 38393 attenuated the amnesic effect of a detention in a dangerous compartment and restored the amnesic memory trace. Possible mechanisms of selective involvement of D1 receptors in retention of memory trace of aversive stimuli during extinction and amnesia in mice with different stereotypes of behavior are discussed. PMID- 16217970 TI - [Correlated activity of neurons in the sensorimotor cortices of the left and right hemispheres of rabbit brain during immobilization catatony]. AB - Spike trains of individual neurons in the sensorimotor cortex detected in simultaneous multiunit records from the left and right rabbit brain hemispheres were analyzed in the baseline state, during immobilization ("animal hypnosis"), and after the termination of this state. The crosscorrelation analysis of pulse series revealed a relationship between the moments of spike generation by neurons of the left and right hemispheres. A significant correlation between the spike trains was considered as an interaction between the neurons (i.e., mutual influence of the cells on each other). It was shown that the strength of the effects of left-hemispheric neurons on cells of the right hemisphere could significantly vary (as compared to baseline) at any stage of the experiment and in any of the time periods analyzed. The strength of the effects activity by neurons of the right-hemispheres on cells of the left-hemispheres significantly changed only after the termination of the immobilization state and in substantially lower time limits. PMID- 16217971 TI - [Avoidance conditioning and morphometric characteristics of the sensorimotor cortex of rats exposed to social deprivation in early ontogeny]. AB - Male Wistar rats were exposed to social deprivation in the period from the 22nd to 70th days of postnatal development. The characteristics of the learning of these rats at the age 110-120 days in the passive avoidance box and during training for active avoidance of electroshock were significant lower than those of control animals. Features of motor activities of the "isolants" in learning and testing, and in the "open field" may be related to their higher anxiety level. Morfometric studies of neurons and glial cells and neocortex thickness showed that significantly lower density of neural elements may be the result of the afferent sensory input limitation produced by the social deprivation. PMID- 16217973 TI - The right to life versus the right to live. PMID- 16217972 TI - [Significance of initial emotional state for neuroimmunomodulation under conditions of activation or blockade of 5-HT1A receptors]. AB - Selective agonist of 5-HT1A receptors--8-OH-DPAT (1 mg/kg) induced a suppression of the immune reaction in aggressive male CBA mice immunized with SRBC (5 x 10(8)). In submissive mice with 10-day defeat experience in confrontation tests, the activation of 5-HT1A receptors with 8-OH-DPAT did not alter the immune response, whereas the application of selective antagonist of 5-HT1A receptors WAY 100635 increased the immune reaction only in submissive mice. It is concluded that activation or blockade of 5-HT1A receptors produced different effects on the immune function of CBA mice depending on the initial emotional state which is known to be provided in aggressive and submissive animals by different activities of the brain neurotransmitter systems including the 5-HTergic system. PMID- 16217974 TI - Imiquimod. PMID- 16217975 TI - Hematogenous vertebral osteomyelitis due to Staphylococcus aureus in the adult. PMID- 16217976 TI - Hematogenous vertebral osteomyelitis due to Staphylococcus aureus in the adult: clinical features and therapeutic outcomes. AB - OBJECTIVE: Staphylococcus aureus is the most common cause of hematogenous vertebral osteomyelitis in adults. To better define clinical features and therapeutic outcomes, the charts of 40 adult patients with S aureus hematogenous vertebral osteomyelitis were retrospectively reviewed. METHODS: Retrospective chart review using standardized data collection form. RESULTS: S aureus hematogenous vertebral osteomyelitis commonly occurred in the settings of recent invasive procedures (55% of patients), insulin use (28%), and hemodialysis (20%). Ten percent of patients had S aureus bacteremia or vascular catheter infection within the preceding 6 months. Median time from first symptom to diagnosis was 51.3 days. A portal of entry for S aureus was identified in 13 patients (32.5%); intravenous catheters were the likely origin in 9 of those 13 patients. Concurrent endocarditis was present in 4 patients. Forty-eight percent of patients had neurologic abnormalities and 60% of patients had an epidural, paraspinous, or psoas abscess demonstrated by neuroimaging. S aureus was isolated through fine-needle aspiration in 17 of 23 patients (74%) and from blood cultures in 23 of 34 patients (68%). Infection was due to methicillin-susceptible S aureus in 67.5% of patients. All patients received intravenous antibiotics for a mean duration of 58.6 days; 36 of 40 (90%) also received concomitant rifampin. Twenty seven percent and 12.5% of patients underwent surgical debridement and CT-guided drainage of abscesses, respectively. After intravenous therapy, 19 of 30 eligible patients received oral continuation treatment. The mean duration of total antibiotic therapy was 142.2 days. CONCLUSIONS: Cure of infection was achieved in 83% (24/29) of evaluable patients, but 50% of those achieving cure still had infection-related sequelae. Intravenous antibiotic therapy for at least 8 weeks was the only clinical factor associated with cure (P = 0.05, two-tailed Fisher exact test). PMID- 16217977 TI - Telithromycin for the treatment of acute bacterial maxillary sinusitis: a review of a new antibacterial agent. AB - OBJECTIVE: Telithromycin, the first approved ketolide antibiotic, was developed to treat community-acquired respiratory tract infections, including acute bacterial maxillary sinusitis (ABMS). A previously published study showed that a 5-day course of 800 mg telithromycin once daily is as effective as a 10-day course in the treatment of ABMS. MATERIALS AND METHODS: Data were pooled from two controlled, multinational, prospective, randomized, double-blinded ABMS trials comparing 5-day telithromycin (800 mg once daily) with 10-day amoxicillin clavulanate (500/125 mg 3 times daily) and cefuroxime axetil (250 mg twice daily). Clinical cure and bacteriologic eradication rates were compared by means of descriptive statistics. RESULTS: The clinical cure rate for telithromycin was 80.9% versus 77.4% for comparators; bacteriologic eradication rate for telithromycin was 84.9% versus 81.7% for comparators. Most adverse events were mild to moderate in intensity and, most commonly, gastrointestinal in nature. CONCLUSIONS: These results support the conclusion that 5 days of treatment with telithromycin is as safe and effective in patients with ABMS as a 10-day course of treatment with amoxicillin-clavulanate or cefuroxime axetil. PMID- 16217978 TI - Use of complementary and alternative medicine by parents of children with HIV infection and asthma and well children. AB - OBJECTIVE: Although the use of complementary and alternative medicine (CAM) in children is increasing, little is known about its use in children with human immunodeficiency virus (HIV) infection. This study investigated the use of CAM by parents of children with HIV infection (H), children with asthma (A), and well children (W). Our hypothesis was that parents of the H group are more likely to use CAM than parents of the W or A group because of the nature and severity of the illness. MATERIALS AND METHODS: Parents of 152 subjects [H (n = 46), A (n = 53), and W (n = 53)] were interviewed on the use of CAM for their children, types of CAM, reasons for use/nonuse, methods of payment, and perceived benefits for their children. RESULTS: Compared with parents of the W and A groups, parents of the H group were less likely to be employed, were less likely to have private insurance, were less likely to have a high school or college education, and were more likely to be black. Interestingly, 38% of the W parents used CAM in their children compared with 22% in the H group and 25% in the A group. More than 80% of all three groups paid out of pocket for their use of CAM in their children. Within these groups, H parents were more likely to want CAM as part of their child's medical care (H = 91% vs W = 75% and A = 67%, P = 0.02) and were more likely to believe that CAM was expensive (H = 78% vs W = 57% and A = 60%, P < 0.01). CONCLUSIONS: Our study revealed a relatively high rate of CAM usage by parents of all three study groups. Although parents of children with HIV infection were more likely to want CAM as part of their children's medical care, their rate of CAM usage was not higher than that in well children. This may be related to their socioeconomic factors. A larger and more diverse study population may provide more information on factors contributing to CAM usage in chronically ill and well children. PMID- 16217979 TI - Pre-event willingness to receive smallpox vaccine among physicians and public safety personnel. AB - BACKGROUND: Planning for voluntary smallpox vaccination of health and safety officials began in December 2002. MATERIALS AND METHODS: Surveys were conducted among physicians and fire and police department personnel in Atlanta, Georgia. Information on demographics, willingness to receive smallpox vaccine, self reported knowledge level, and potential vaccine contraindications was analyzed. RESULTS: Forty-one percent of physicians (n = 199) were undecided on vaccination (32% would receive vaccine and 27% would not). Forty-eight percent of firefighters (n = 343) and 41% of police (n = 466) were undecided; 23% and 41% would receive vaccine, whereas 28% and 18% would not (fire and police, respectively). Absence of contraindications was associated with physicians' willingness to be vaccinated (P = 0.006). Many physicians (66%) and most public safety personnel (88%) considered themselves inadequately informed on smallpox vaccine. In a multivariate analysis, inadequately informed respondents were more likely to be undecided (OR = 2.23, CI = 1.39 to 3.56). CONCLUSIONS: Before implementation of the smallpox vaccination program, self-assessed knowledge about smallpox disease and vaccine were poor. PMID- 16217980 TI - Does physician benchmarking improve performance of laparoscopically assisted vaginal hysterectomy? AB - BACKGROUND: Benchmarking techniques were implemented to optimize operating time and charges associated with laparoscopically assisted vaginal hysterectomy (LAVH). MATERIALS AND METHODS: The baseline LAVH profile over a period of 4 years (167 cases) was compared with 1-year data (47 cases) after a benchmarking educational program (disseminating data ranking performance by each surgeon plus suggestions for improvement). Preintervention and postintervention profiles were compared by means of Student t test and wilcoxon rank sum analysis. Hierarchical multiple regression was used to identify additional sources of variation for operative charges and time. RESULTS: Mean operating times after implementing benchmarking were lower, averaging 182 versus 197 minutes in the control subjects (P = 0.05). We found no significant difference in total or operative charges. After adjusting for potential confounders, benchmarking remained associated with decreased operating time in the multivariate model (P = 0.01). CONCLUSIONS: LAVH operating times decreased after a surgical benchmarking and education intervention, but operating charges did not. PMID- 16217981 TI - Knowledge of the principles of judicious antibiotic use for upper respiratory infections: a survey of senior medical students. AB - OBJECTIVE: Senior medical students (n = 2,433) from 21 accredited medical schools in New England and the mid-Atlantic states were surveyed to evaluate their knowledge of and compliance with principles of judicious antimicrobial use, as defined by the Centers for Disease Control and others. MATERIALS AND METHODS: A self-administered questionnaire with six vignettes on the clinical management of different upper respiratory tract infections was used. Compliance was calculated by using an ordinal response scale (1 to 4) for each question on the six vignettes. RESULTS: The adjusted response rate was 46%. More than 47% of the respondents had read none of the principles, and only 2.9% had read all six. Approximately 36% of the respondents would start antibiotics within 2 days of an 18-month-old presenting with purulent rhinitis, whereas 55.9% would immediately prescribe antibiotics if the child had wheezy bronchitis. For a 4-year-old with pharyngitis, 29.5% of respondents would either give an antibiotic office sample to start that night and a prescription for continuation of treatment at home, would give an antibiotic prescription with instructions to discontinue treatment with a negative throat culture, or would treat without a throat culture. Almost all of the respondents (99%) were informed regarding the problems of antibiotic resistance, usually from multiple sources. The number of sources of knowledge about problems of antibiotic resistance was the only predictor of compliance (P = 0.02). The number of principles read was not correlated with compliance. CONCLUSIONS: Among students surveyed, large gaps remain regarding the appropriate use of antimicrobial agents for the treatment of upper respiratory infections. PMID- 16217982 TI - Surgical treatment of acute type II and rostral type III odontoid fractures managed by anterior screw fixation. AB - OBJECTIVES: In the present study, the authors comment on their experience with anterior odontoid screw fixation in the management of odontoid fractures, in an attempt to further assess the safety and the efficacy of this procedure. MATERIALS AND METHODS: A retrospective analysis of 50 consecutive patients with reducible type II or rostral type III odontoid fractures, operated at our hospital with anterior odontoid screw fixation. Radiographic bony fusion, complications, and clinical outcome were evaluated. RESULTS: Solid bony fusion was evident in 38 (90.5%) of the patients. One mechanical instrumentation-related complication occurred, without clinical significance. No other major complications related to the procedure were noted. A satisfactory range of motion in the cervical spine was observed in all patients. CONCLUSIONS: Anterior odontoid screw fixation is a safe and effective procedure for the treatment of type II and rostral type III odontoid fractures. Compliance to the specific indications and contraindications of this operation is crucial for optimal outcome. PMID- 16217983 TI - Insulin resistance in polycystic ovarian disease. AB - The classic polycystic ovarian syndrome (PCOS) was originally described by Stein and Leventhal as the association of amenorrhea with polycystic ovaries and, variably, hirsutism and/or obesity. It is estimated that 5 to 10% of women of reproductive age have PCOS. Although insulin resistance is not part of the diagnostic criteria for PCOS, its importance in the pathogenesis of PCOS can not be denied. PCOS is associated with insulin resistance, independent of total or fat-free body mass. Postreceptor defects in the action of insulin have been described in PCOS that are similar to those found in obesity and type 2 diabetes. Treatment with insulin sensitizers, metformin, and thiazolidinediones (TZDs) improve both metabolic and hormonal patterns and also improve ovulation in PCOS. Recent studies have shown that women who have PCOS have higher circulating levels of inflammatory mediators such as C-reactive protein, tumor necrosis factor, tissue plasminogen activator, and plasminogen activator inhibitor-1 (PAI-1). It is possible that the beneficial effect of insulin sensitizers in PCOS may be partly due to a decrease in inflammation. PMID- 16217984 TI - Current and potential uses of imiquimod. AB - Imiquimod, an imidazoquinoline amine, is an immune response modifier first FDA approved for the treatment of external genital and perianal warts in 1997. Since its appearance on the market, its antiviral and antitumor properties have been used in the treatment of a variety of dermatologic conditions. In this review article, the basic mechanism of action of imiquimod, current FDA-approved and non FDA-approved uses of imiquimod, and key points of medication application frequency, possible adverse effects, and use in combination therapy are discussed. Common skin conditions that may be eradicated with imiquimod are emphasized. PMID- 16217985 TI - Group C streptococcal sepsis complicating Fournier gangrene. AB - Fournier gangrene is a life-threatening necrotizing fasciitis of the perineal scrotal area that occurs in diabetic males. Patients typically present with systemic toxicity and significant inflammatory changes in the scrotum and perineum. Most cases of Fournier gangrene are polymicrobic and require urgent surgical debridement and broad-spectrum antibiotic therapy. We describe a case of Fournier gangrene in a young diabetic man that was associated with group C streptococcal bacteremia, an association previously unreported in the literature to our knowledge. PMID- 16217986 TI - Mycobacterium tuberculosis infection of a presumed Charcot joint. AB - A 65-year-old male with peripheral neuropathy and small lymphocytic lymphoma presented with erythema and edema of the left foot. A Charcot midfoot was diagnosed and treated with a total contact cast and restricted weight bearing. However, subsequent analysis of bone and synovial fluid months later revealed Mycobacterium tuberculosis infection. PMID- 16217987 TI - Unusual endoscopic and microscopic view of Enterobius vermicularis: a case report with a review of the literature. AB - Enterobius vermicularis has the broadest geographic range of any helminth and is the most common intestinal parasite seen in the primary care setting. Underappreciated is the fact that it is not always a benign disorder and could even cause life-threatening medical problems. Visualization of the actual worms during endoscopy is probably underappreciated in part because endoscopists have never actually seen the worm and/or are not actively looking for, or anticipating, worms. This report describes a case of worm infection as documented during colonoscopy and confirmed by microscopy. The gross and microscopic appearance of the worm is described. Literature regarding the wide range of gastrointestinal and nongastrointestinal manifestations, including potentially life-threatening illnesses, as well as treatment options, are also reviewed. PMID- 16217988 TI - Coccidioidomycosis diagnosed in South Carolina. AB - Primary care physicians in the Eastern United States rarely consider coccidioidomycosis in the differential diagnosis of pulmonary infections or febrile illnesses. However, the mobility of the population mandates consideration of this diagnosis, particularly in patients with fever and cough that do not resolve rapidly and in patients with adenopathy on chest radiography. In this report, we describe two unrelated cases encountered during a single week in a South Carolina internal medicine practice. These cases highlight the importance of obtaining travel histories from patients with atypical pulmonary infections. Early consideration of coccidioidomycosis confers several benefits, including allaying patient anxiety by more timely diagnosis, minimizing the empiric use of antibiotics, and reducing the need for extensive and possibly invasive diagnostic testing. PMID- 16217989 TI - Kaposi sarcoma secondary to pulmonary tuberculosis: a rare case. AB - Kaposi sarcoma commonly occurs in HIV-positive and immunocompromised patients. We describe a case of Kaposi sarcoma that developed in an HIV-negative patient with tuberculosis. The Kaposi sarcoma completely regressed with antituberculous therapy without the institution of chemotherapy. The patient remained disease free after a follow-up period of 20 months. Patients with Kaposi sarcoma should be monitored for coexisting diseases such as tuberculosis. PMID- 16217990 TI - Sarcoidosis mimicking disseminated testicular cancer. AB - Sarcoidosis rarely involves the genitourinary system or the spinal cord. This report describes a case of sarcoidosis with a scrotal mass, a mass compressing the spinal cord, and lesions in the liver and spleen. This case illustrates an uncommon manifestation of sarcoidosis, which may mimic disseminated testicular cancer. PMID- 16217991 TI - Necrotizing lymphadenitis associated with the phenytoin-induced hypersensitivity syndrome. AB - A 32-year-old black female was started on phenytoin for seizure prophylaxis following the clipping of an aneurysm. This was stopped after 3 weeks when she developed a generalized skin rash. Over the next week she developed fever, sore throat, dysphagia, and headache. She had an erythematous throat with white exudates on the right tonsil and 1 to 3 cm firm, tender lymphadenopathy in multiple regions. Blood, throat swab and cerebrospinal fluid studies were negative for bacterial or viral infections, except for elevated liver enzymes. CT scan of chest, abdomen, and pelvis showed no lymphadenopathy. Lymph node biopsy suggested necrosis but no evidence of infection, granuloma, or lymphoma. Her lymphadenopathy resolved spontaneously and liver enzymes normalized in 3 weeks. Hypersensitivity syndrome due to antiepileptics manifests as fever, rash, generalized lymphadenopathy, and probably represents a T-cell mediated drug reaction. This reaction may persist despite cessation of the drug, and it may engender expensive evaluation. Careful observation up to 3 weeks after drug cessation may be the best management. PMID- 16217992 TI - Beneficial effect of immunosuppressive drugs on Parry-Romberg syndrome: a case report and review of the literature. AB - Progressive facial hemiatrophy, also known as Parry-Romberg syndrome (PRS), is characterized by slowly progressive atrophy of one side of the face, primarily involving the subcutaneous tissues and fat. Involvement of the central nervous system with impairment of neurologic function occurs infrequently. At present, there is no agreement as to whether PRS is a distinct entity or a clinical variant of linear scleroderma en coup de sabre. The exact reason for PRS has not yet been determined; therefore, no suitable treatment exists. We observed beneficial effects of immunosuppressive agents on neurologic lesions in particular in a patient with PRS who presented with immunoinflammatory findings and neurologic involvement, apart from cutaneous manifestations. PMID- 16217993 TI - Left ventricular endocardial calcification in a patient with myeloproliferative disease. AB - The case of a 57-year-old male with a history significant for myeloproliferative disease, chronic renal failure, hypertension, and prostate cancer is described. His complete blood count was remarkable for neutrophilia and, notably, eosinophilia. Subsequent to two syncopal episodes, a transthoracic echocardiogram was performed as part of the workup, which showed an unusual calcified mass in the left ventricular apical region but separate from the apical myocardium, with normal left ventricular systolic function. A transesophageal echocardiogram and computed tomography of the chest confirmed the presence of extensive calcification in the left ventricle of unusual location and shape. This patient probably had Loeffler endocarditis related to myeloproliferative disorder, complicated by calcification of the endocardial sclerotic lesions. PMID- 16217994 TI - Four primary tumors of lung, bladder, prostate, and breast in a male patient. AB - We present a very rare case of quadruple cancers in a 65-year-old male patient. It is a case of both synchronous and metachronous primary malignant neoplasms occurring in four different organs. Immunohistochemical stains showed tumor cell nuclei to be negative for p53 over-expression. To our knowledge, this is the first documented case with this combination of primary tumors. The tumors included an adenosquamous cell carcinoma of the lung, transitional cell carcinoma of the urinary bladder, and adenocarcinomas of the prostate and the breast. We also review the medical literature for the possible causes of multiple primary malignant neoplasms. PMID- 16217995 TI - Reversible ST-segment elevation associated with atelectasis of the left lung. AB - A 22-year-old male with cerebral palsy and respiratory failure had acute reversible ST-segment elevation in the inferior leads during acute collapse of the left lung, which resolved with re-expansion of the left lung several hours later. This suggests that major lung actelectasis needs to be added to the group of noncardiac conditions considered when evaluating ST elevation in the critically ill patient. PMID- 16217996 TI - Substance-abuse slang terms and implications for cultural groups including Orthodox Jews. PMID- 16217997 TI - Cast abscess. PMID- 16217998 TI - Fatal Trichosporon asahii septicemia in a Guatemalan farmer with acute lymphoblastic leukemia. PMID- 16217999 TI - Unintentional carbon monoxide poisoning cases in Ciudad Juarez, Mexico. PMID- 16218000 TI - Response to type 2 diabetes: epidemiologic trends, evolving pathogenic concepts, and recent changes in therapeutic approach. PMID- 16218001 TI - Pleural effusion as presentation of metastatic adenocarcinoma of prostate. PMID- 16218002 TI - The patient's page. PMID- 16218003 TI - Evaluation: Part I: Evaluating learning activities. PMID- 16218004 TI - Targeted therapies in the treatment of cancer. PMID- 16218005 TI - Keeping participants enrolled: use of gradient. PMID- 16218006 TI - Ethical leadership. PMID- 16218007 TI - Outcomes-based research: evaluating the effectiveness of an online nurse refresher course. AB - According to the Monthly Labor Review, 1 million new nurses will be needed by the year 2010. Returning inactive nurses to the profession is undoubtedly an economical and cost-effective approach to address this need. Embracing an online methodology will reach nurses in less populated, geographically dispersed areas with limited access to traditional educational opportunities. On the basis of the principles of good practice, it is imperative that educators evaluate the effectiveness of distance methodology. This article provides empirical data supporting the online learning process and consumer satisfaction of an online nurse refresher course. PMID- 16218008 TI - Hong Kong nurses' perceptions of and participation in continuing nursing education. AB - As medical technologies change, practicing nurses need to update their knowledge and skills to ensure quality health care for patients and to minimize possible health hazards in the workplace. This article describes a study that explored Hong Kong hospital nurses' perceptions of and participation in continuing nursing education. It found that Hong Kong nurses participate actively in continuing education out of a sense of professional responsibility and personal interest. However, consistent with findings from other studies, the major factors hindering nurses' participation are finances, family commitments, and time. PMID- 16218009 TI - Registered nurse refresher course as an adjunct in nurse recruitment. AB - A registered nurse refresher course in the east central region of Missouri has offered a singular opportunity for nurses wishing to re-enter the nursing profession. By offering both didactic instruction and clinical experience with a nurse mentor in the medical center, nurses who have been out of nursing or in a non-clinical role have had the chance to begin to reorient themselves to professional practice before committing to a permanent position. This article describes one institution's experience and process for those who might be considering developing a re-entry program of their own. PMID- 16218011 TI - Innovative ways to promote recruitment, retention, collegiality, and professionalism using continuing education plans. AB - Technological advances in health care, coupled with the crisis of a nursing shortage, make recruitment and retention of well-educated nurses a key factor for the viability and growth of any healthcare organization. In the past, symbiotic alliances have been formed solely between healthcare organizations and nurses. Professional nursing organizations were rarely involved in the alliance. Using the Magnet Recognition Program, one local chapter of Sigma Theta Tau International has found a way to unite nurses, healthcare organizations, and professional nursing organizations to encourage nursing education, promote positive patient outcomes, and help find a solution to the problem. PMID- 16218010 TI - Attention and restoration in Post-RN students. AB - BACKGROUND: The impact of a restorative intervention using the natural environment on capacity to direct attention and issues that contribute to attention fatigue for diploma-prepared nursing students (Post-RN students) enrolled in a baccalaureate nursing program was examined. METHODS: This study used a quasi-experimental comparison group design. Subjective (Attentional Functional Index) and objective (Finding A's Test and Symbol Digits Modalities Test) measures were employed. RESULTS: Thirty-two students at two universities participated. Results of the split-plot analysis revealed a within-subject effect on the Attentional Functional Index (p < .05), a significant within-subject effect on the Finding A's Test (p < .05), and a significant within- and between subject effect on the Symbol Digits Modalities Test (p < .05). CONCLUSION: Recognizing and managing attention fatigue throughout the school year may enhance Post-RN students' abilities to direct attention and contribute to an enhanced academic experience. PMID- 16218012 TI - Continuing education for nurses: a necessity or a nicety? AB - The changing health care climate has resulted in escalating financial constraints. One department that is often scrutinized for its cost-benefit relation to the hospital is the nursing education department. These departments are increasingly being reduced in size and investment in the continuing education of nurses is being rationalized. However, reducing financial support of education may be counterproductive in both the short- and long-term. This article does not propose a "recipe" for effectively facilitating continuing education. However, it does provide sound justification for investing in the continuing education of nurses and demonstrates not only that quality education results in enhanced knowledge and skills, but that there is also a positive correlation between professional development and factors such as staff satisfaction, staff retention, and quality patient care. PMID- 16218013 TI - British health care and imaging. PMID- 16218015 TI - Health language skips class. PMID- 16218014 TI - Sonographic transducer instrumentation. PMID- 16218016 TI - Educating the hybrid technologist. PMID- 16218017 TI - Tangential projection of the patella. PMID- 16218018 TI - RA certification program. PMID- 16218019 TI - Teaching critical-thinking skills through group-based learning. AB - CONTEXT: Critical-thinking skills require the radiographer to analyze, evaluate and synthesize situations to determine a course of action most beneficial to the patient. OBJECTIVE: For radiography educators, it is important to understand how critical-thinking skills are acquired during the educational process for each student to reach his or her maximum potential. METHOD: Various teaching strategies such as problem-based learning are reviewed and discussed in this article. Although the teaching strategies discussed have been used in medical education for over 40 years, they are fairly new to radiography educational programs. CONCLUSION: Today's educators must prepare graduates who are proficient in a wide range of skills to work in a variety of clinical settings. This requires the integration of group-based learning into the radiography curriculum. PMID- 16218020 TI - Radiation dose in pelvic imaging. AB - OBJECTIVE: To compare the radiation dose during pelvic x-ray examinations using computed radiography (CR) and film-screen (FS) radiography at various x-ray tube voltages (kV) and tube-current time product (mAs) values. METHODS: A pelvic phantom was imaged using FS and CR systems. The entrance surface dose was measured using an ionization chamber, and the gonadal dose and effective dose were calculated using the XDOSE program. The diagnostic quality of the images was assessed using a 5-point subjective scoring system. RESULTS: At standard kV values, the image quality did not vary significantly between the CR and the FS system, but at higher kV values, the CR images werefound to be of better quality than FS images. In addition, the lower limit of entrance skin dose consistent with diagnostically acceptable CR images was 50% lower than that for FS images. CONCLUSION: The gonadal dose and effective dose for pelvic x-ray examinations can be reduced by 50% when CR systems are used and appropriate exposure factors are established. PMID- 16218021 TI - Aortic emergencies. AB - This article discusses the anatomy and pathobiology of the aorta, common aortic emergencies and their diagnosis, and the roles and limitations of diagnostic imaging in assessment, medical intervention and monitoring. PMID- 16218022 TI - Traumatic injuries to the cervical spine. AB - Nowhere in radiography are the standards for imaging more important than in dealing with traumatic spine injury. This article provides an overview of cervical spine trauma and specifically addresses various injuries and how to image them. PMID- 16218023 TI - Oval, egg or evidence? PMID- 16218024 TI - Patient page. If you are from another country. PMID- 16218025 TI - Innate immunity processes in organ allografting--their contribution to acute and chronic rejection. AB - The innate immune processes proceeding in organ allografts and their contribution to acute and chronic rejection have been described. Ischemia/reperfusion phenomenon with damage of the endothelial and parenchymal cells and reaction of granulocytes and macrophages as well as humoral factors including complement, coagulation factors, free radicals, nitrous oxide have an impact on late function of the transplanted organs. Participation of granulocytes and macrophages in acute allogeneic rejection is another example of the role of the innate system in transplant damage. Recurrent basic disease, diabetes, hyperlipidemia, adverse effects of immunosuppressive drugs in allograft recipients are factors evoking reaction of the innate system. Investigations of the innate immune system have shown an essential role, beside of the adaptive system stimulated by allogeneic mismatch, in organ graft rejection. PMID- 16218026 TI - Bone contamination and blood culture in tissue donors. AB - INTRODUCTION: Swab cultures are the most usual method to detect graft contamination; nevertheless it has been confirmed his limited sensibility. We have studied the relationship between blood cultures, swab surface cultures and cultures of entirely samples of cancellous bone. MATERIAL AND METHODS: We have evaluated 5 donors with positive blood culture, from 70 multiorganic donors during 2002. Blood samples were obtained prior the heart arrest. The bone procurement was done just after the organ recovery under aseptic conditions, and surface cultures were performed of each bone. After storage at -80 degrees C, cancellous samples were obtained by trephine and were completely cultured. RESULTS: In one case, the same microorganism grown in blood culture, in 2 of 9 surface cultures, and in 15 of 26 samples of cancellous bone. CONCLUSION: We conclude that to guarantee allograft's safety it is recommended to add donor's blood culture to the habitual surface swab culture if secondary sterilisation is not performed. PMID- 16218027 TI - Increased plasma TGF-beta 1 level in patient with salivary gland carcinoma after renal transplantation. AB - TGF-beta1 maintains homeostasis of rapidly proliferating cells, therefore is a potential tumor suppressor of non-malignant cells. Malignant transformation alters TGF-beta1 signalling pathway, turning it into a stimulator of tumor progression. We describe the case of 54 year old renal transplant patient with highly increased plasma TGF-beta1 level and planoepithelial carcinoma originating from parotid salivary gland. We discuss the role of TGF-beta1 in promotion of carcinogenesis and probable utility in prognosis of malignancy. PMID- 16218028 TI - Non-heart-beating donors: is it worthwhile? AB - OBJECTIVE: Due to organ shortage in renal transplantation, many transplant centers attempt to increase the donor pool. Non-heart-beating donors seem to be a promising alternative. PATIENTS AND METHODS: We performed 14 renal transplantations from 8 non-heart-beating donors. 2, 1, 3 and 2 donors were from groups 1, 2, 3 and 4, respectively, according to Maastricht classification. RESULTS: In 6 of the patients' warm ischemia time was over 30 minutes. Three of them had primary non-function. In 6 patients delayed graft function was seen. The remaining 5 kidneys functioned immediately. Two patients whose kidney functioned returned to hemodialysis in the I I and 13th months after their transplantations. One of the patients with primary non-function died. 9 kidneys function well in their follow-up period between 5-111 months. 1 and 5-year graft survival rates were 69.8 and 61.1 percent, respectively. The mean graft survival time is 69.9 +/ 14.5 months. CONCLUSION: Despite the high primary non-function rate, we think that non-heart-beating donors especially in Maastricht classification 3 and 4 should be used due to dramatic shortage of organs. PMID- 16218029 TI - Monitoring of human cytomegalovirus, HHV-6 and HHV-7 infection in kidney transplant recipients by molecular methods to predict HCMV disease after transplantation: a prospective study. AB - OBJECTIVES: Recently, highly sensitive molecular assays to detect HCMV, HHV-6 and HHV-7 have been developed but their ability to detect patients at high risk for disease is unclear. METHODS: The positive predictive values (PPV) of pp65 antigenemia, quantitative plasma DNA and pp67-mRNA for CMV-disease were prospectively compared in 82 transplant recipients (72 renal, 10 pancreas-kidney) without CMV-prophylaxis. In addition, the prevalence of HHV-6 and HHV-7 infection were assessed using qualitative PCR. The assays were performed weekly. RESULTS: Three patients (3,7%) developed CMV-disease and were effectively treated. They were positive in all three CMV-assays. The PPVs of pp65-Ag, DNA viral load and pp67-mRNA were 33%, 20% and 25% in CMV-positive and 100%, 67% and 50% in seronegative recipients. Sensitivity and negative predictive value were 100% for all assays. Using cut-offs, PPVs were 75% (pp65-Ag > or = 20/200.000 cells) and 100% (PCR > or =30.000 copies/ml). Transfusion of >2 packed red cells, rejection and non-functioning graft were risk factors for CMV Five patients and one patient were positive for HHV-6 and HHV-7 resp.; both were symptomless and did not have a HCMV infection. CONCLUSIONS: Therefore, pp65-antigenemia and plasma PCR with a cut-off could be useful for monitoring preemptive therapy. PMID- 16218030 TI - The impact of immunosuppressive therapy on an early quantitative NK cell reconstitution after allogeneic haematopoietic cell transplantation. AB - OBJECTIVES: The goal of this study was to examine the impact of various immunosuppressive regimens on an early NK cell recovery following haematopoietic cell transplantation (alloHCT). METHODS: The number of peripheral blood NK cells was analysed with the use of flow cytometry on day +30 (+/-2) after alloHCT from an HLA identical sibling (n=43) or an unrelated (n=34) donor. RESULTS: Patients receiving prednisolone as a prophylaxis of acute graft-versus-host disease had lower number of NK cells compared to those not given steroids prophylactically (110(10-694) vs. 212(33-890) x 10(6)/L, p = 0.005). In contrast, administration of anti-thymocyte globulin (ATG) (7.5-15 mg/kg) as a part of preparative regimen was not found to influence the NK cell recovery. Similarly, no effect on the number of peripheral blood NK cells was observed with regard to other analysed factors: cell dose, type of myeloablative conditioning, source of stem cells, patient and donor characteristics, number of post-transplant methotrexate doses. CONCLUSIONS: Immunosuppressive therapy may affect NK cell recovery following alloHCT. Since NK cells are considered a potential tool for cellular therapy of haematological malignancies, our findings should be taken into account when planning this kind of treatment in the context of allotransplantation. PMID- 16218031 TI - Multiple renal arteries in laparoscopic donor nephrectomy. AB - BACKGROUND: Laparoscopic donor nephrectomy has gained popularity throughout the world recently. The more centers became experienced the more this technique began to be used, even in extreme cases. Kidneys with multiple renal arteries are one of the difficult cases for laparoscopic donor nephrectomy. PATIENTS AND METHOD: Thirty living laparoscopic donor nephrectomies have been performed between January 2001 and December 2002. Twenty-three of them had single and seven kidneys had multiple renal arteries. Single (SA) and multiple (MA) artery groups were statistically similar in terms of donor age, rate of received right kidneys and serum creatinine clearance of the donors. RESULTS: The mean duration of the donor surgery was 225 min and 240 min in SA and MA groups. In the SA and MA groups, the mean warm and cold ischemia times were statistically similar. Laparoscopic nephrectomy was converted to open procedure in five and one donors, in the SA and MA artery groups, respectively. Intraoperative bleeding (single artery: 4, multiple arteries: 1) was the most common cause for conversion. Postoperative urinary complications were seen in four and one patients, in the SA and MA groups, respectively. Lengths of hospital stay of the donors were similar in both groups. Serum creatinine levels of the patients on seventh, 30th, 90th days, and 1 year were found to be statistically similar in both groups. CONCLUSION: Laparoscopic donor nephrectomy was found to be a safe procedure in the donors with multiple renal arteries by the experienced surgical teams, as much as in donors with single arteries. PMID- 16218032 TI - Clinical application of monitoring mycophenolic acid trough concentration in heart transplant recipients--single center's experience. AB - The purpose of this study was to assess the clinical utility of mycophenolic acid (MPA) trough concentration monitoring in heart transplant recipients. METHODS: We reviewed 456 MPA plasma level measurements (EMIT/Dade-Behring) which were performed in 76 pts. after orthotopic heart transplantation (OHT): 57 M and 21 F, age 41.9 +/- 16, time after OHT (months) 17.6 +/- 24. Daily dose of mycophenolate mofetil (MMF) was 2-3 g before MPA measurement introduction, then it was adjusted to achieve MPA trough levels (TL) of 2-4 microg/ml. Additionally pts. received either cyclosporine-A (CyA) or tacrolimus and prednisone. We analyzed first MPA levels obtained in pts. without previous monitoring, then we looked for a relation between CyA or tacrolimus and MPA level, and finally we checked for a relation between MPA level and the side-effects of MMF RESULTS: In a group of 59 pts. without earlier MPA level monitoring we found that 36 pts. (61%) had MPA level below, 19 pts. (32%) within, and 4 pts. (7%) above target TL. We identified a group of 11 pts. (characterized by unstable CyA TLs and liver impairment) with a significant positive correlation between CyA and MPA level. For the remaining group of pts. we found a non-significant negative correlation between CyA and MPA concentrations. Target MPA TL was achieved in 40% of cases in pts. with CyA TL below 200 ng/ml (Axsym/Abbott), in 40% of cases in pts. with CyA TL 200-300 ng/ml, and in 27% of cases in pts. with CyA TL over 300 ng/ml. There was no correlation between tacrolimus and MPA level. MPA TL over 4 microg/ml occurred in 22% of results from pts. receiving tacrolimus (n=6) and 11% of pts. on CyA (n=17, p = 0.011). 90% of these pts. had symptoms of GI irritation, 33%--leucopoenia, and 14%--anemia. CONCLUSIONS: It is uncommon to achieve MPA TL of 2-4 microg/ml with typical doses of MMF, especially with concomitant high CyA TL. Typical side effects of MMF should be an indication to check MPA TL. PMID- 16218033 TI - Distribution of porcine endogenous retroviruses (PERVs) DNA in organs of a domestic pig. AB - OBJECTIVES: Domestic pig may serve as the most appropriate organ source for human xenotransplantation in the future. However, there is a serious threat of xenogeneic pathogens transmission, especially porcine endogenous retroviruses (PERVs) which are present in genomes of all pigs. The aim of this study was to monitor the prevalence and distribution of PERV DNA in organs of a domestic pig. METHODS: We used a primer set for a highly conserved fragment of PERV gag sequence to monitor a total PERV DNA copy number and genotype-specific primer sets to study PERV subtypes distribution using Real-Time QPCR (SYBR Green I). RESULTS: Our results showed that PERV DNA was present in all studied pigs, however, most PERV DNA molecules carried numerous mutations thus indicating inability to express functional retroviral particles. The level of PERV DNA in kidney was much higher than in heart (p = 0.007) and in the liver (p = 0.009). CONCLUSIONS: It indicates that kidney is potentially the biggest PERV reservoir which makes it the organ of particular concern in xenotransplantation. We also conclude it is possible to monitor pig herds for individuals with the lowest PERV DNA prevalence, especially lacking PERV-C, and perhaps with only defective PERV proviruses that are unable to express functional RNA. PMID- 16218034 TI - Longitudinal study of renal function in pediatric liver transplant recipients. AB - OBJECTIVES: Study was performed to evaluate prospectively long-term renal function in pediatric liver transplant recipients. METHODS: In 50 children aged 1 18 years, renal function tests (sCr, GFR, ERPF, Cr clearance, renal Doppler ultrasound, ABPM) have been performed before and several times during 36 months follow-up after liver transplantation. RESULTS: Signifficant reduction of renal filtration function was found in studied children (increase of sCr, decrease of GFR, ERPF, Cr clearance) within 12 months after transplantation, which did not progress further in most children after this time. None of patients progressed to stage 4 or 5 renal failure according to CKD. Schwartz formula was not found reliable in assessing renal function in children after liver transplantation. There was no difference in renal function according to cyclosporine or tacrolimus treatment, however arterial hypertension was more common in cyclosporine treated group. CONCLUSIONS: Although renal damage may be a problem for an individual chuild after liver transplantation, majority of children have stable renal function 36 months after Tx. DTPA GFR should be performed yearly in all children until adulthood not to overlook progression of renal injury. PMID- 16218035 TI - Kaposi's sarcoma in renal transplant recipients. AB - Kaposi's sarcoma (KS) is a spindle-shaped vascular cell tumor that occurs in the skin, lymphoid, respiratory and gastrointestinal tissues. It may resemble aggressive malignant neoplasm in HIV-related or in post-transplant types but classic form may behave as benign, potentially controllable and reversible hyperplasia. KS lesions from the onset are dispersed and multicentric. KS probability increases in solid organ transplant recipients (approximately 3/1000 patients). KS occurrence is associated with: type and dose of immunosuppression, chronic stimulation by foreign allograft antigens, viral infections (Herpes virus 8), anti rejection and induction therapy, etc. 90% of KS cases appear as dark blue or purplish macular lesions that may form nodular tumors. Histological picture shows networks of spindle shaped cells and vascular spaces surrounded by an endothelial cell layer. There is no uniform schema of KS treatment in renal transplant recipients. Immunosuppression must be reduced to the lowest levels which preserve allograft function. CsA should be converted to mofetil mycophenolate or mTOR-inhibitors. After conversion to MMF regression of KS was observed, although low therapeutic MMF doses seem to be appropriate. Sirolimus seems to inhibit the growth of established vascularized tumors and this effect is best realized with relatively low immunosuppressive doses of drug. PMID- 16218036 TI - Iliac artery stenosis as a cause of posttransplant renovascular hypertension: report of two cases. AB - Stenosis of renal artery (TRAS) or iliac artery proximal to the anastomosis (pTRAS) results in an impairment of kidney graft perfusion, arterial hypertension and in consequence decrease of glomerular filtration. In this report we present two patients with pTRAS and renovascular hypertension after kidney transplantation (KTx), treated successfully with percutaneous transluminal angioplasty (PTA) with stenting. Resistant hypertension, decrease of diuresis and increase of serum creatinine concentration were observed in both presented patients. Also in both cases pTRAS was diagnosed by Doppler ultrasonography and confirmed by angiography. Iliac artery angioplasty with stenting was successfully applied in these patients, resulting in a decrease of blood pressure, restoration of diuresis and a decrease of serum creatinine concentration. These observations confirm that PTA of iliac arteries with stenting is an effective method of treatment in patients with pTRAS. PMID- 16218037 TI - Infections caused by clostridium difficile in kidney or liver graft recipients. AB - OBJECTIVES: Antibiotic-associated diarrhea is defined as otherwise unexplained diarrhea which occurs in association with the administration of antibiotics. The incidence of this diagnosis increases worldwide due to augmentative usage of broad spectrum antibiotics. Clostridium difficile is the most common identifiable pathogen, leading to antibiotic-associated diarrhea. The aim of the article was to describe our own experience in diagnostics and treatment of infections caused by C. difficile in solid organs recipients. METHODS: In the article, retrospective analysis of infections caused by C. difficile that occurred during first six months of 2003 in the Department of Immunology, Transplant Medicine and Internal Diseases of the Medical University of Warsaw was performed. RESULTS: In this period 18 infections in 16 kidney or/and liver graft recipients were diagnosed. Risk factors are considered and clinical manifestation is described. CONCLUSIONS: Proper diagnostics and therapy in this population of patients are proposed and preventive procedures are discussed. PMID- 16218038 TI - Identifying cardiac troponin T. PMID- 16218040 TI - Puberty onset in Northern Italy: a random sample of 3597 Italian children. AB - Entering puberty is one of the most important milestones in life. Studies from around the world have shown that age of pubertal changes onset can vary with race and ethnicity, environmental conditions, geographical location and nutrition. In the last century, the onset of puberty progressively shifted back towards younger ages in several European countries, with a levelling off in the last decades. The aim of our study was to describe the prevalence of secondary sexual characteristics in a group of children living in Northern Italy comparing them with the percentile values published by Tanner in 1976. We enrolled 3496 children drawn from public schools and evaluated height, weight and pubertal stages. The analysis of our data evidenced that the 50th percentile age of puberty onset in both sexes decreased by about 1 yr compared to data published by Tanner. Mean body mass index (BMI) z-score was significantly higher (p = 0.01) in pubertal than in pre-pubertal girls, on the contrary it was higher (p = 0.005) in pre pubertal than in pubertal boys. In conclusion, our study found that girls and boys of our region are beginning pubertal development about 1 yr earlier than Tanner's British population. Taking into consideration the 3rd percentile age for Tanner's breast stage 2 in girls and testicular volume (TV) of 4 ml in boys, the current internationally used cut-off age for precocious puberty, i.e. 8 yr for girls and 9 yr for boys, can be maintained in our population. PMID- 16218039 TI - Gamma probe-guided surgery for revision thyroidectomy: in comparison with conventional technique. AB - Reoperative thyroid surgery may be required in patients who undergo any procedure less than total or near total thyroidectomy. The aim of this study was to investigate advantages of gamma-probe guided revision thyroidectomy (GGRT) over conventional revision thyroidectomy (CRT) in patients with differentiated thyroid carcinoma (DTC). GGRT was assessed according to the TSH values, complication rates and the incidence of carcinoma in residual thyroid tissue. In this randomised prospective clinical trial, 25 patients with differentiated thyroid carcinoma who had previously undergone surgery for benign multinodular goiter were included in the study. GGRT was performed in 11 (44%) patients (Group 1) and CRT in 14 (65%) (Group 2). The intraoperative mean ratio of thyroid activity to background activity (T/B) was detected as 5.1 +/- 1.4 and the mean ratio of thyroid bed activity to background activity after excision (Tbed/B) was 1.3 +/- 0.3, (p < 0.01). Although the incidence of carcinoma in residual thyroid tissue was higher in group 1 (4/11) in comparison to group 2 (1/14), it was not statistically significant. The elevation of the TSH concentration at the first post-operative month was significantly higher in group 1 in comparison with group 2 (18 +/- 25 5 +/- 3 mlU/l), (p < 0.02). These results indicate that intraoperative gamma probe application may be beneficial to detect and remove residual thyroid tissue in revision thyroidectomy. PMID- 16218041 TI - Effect of 10 yr of the iodine supplementation on the hearing threshold of iodine deficient schoolchildren. AB - OBJECTIVE: Auditory disturbances may be present in iodine deficient children. The aim of this study was to determine the effect of long-term iodine supplementation on auditory thresholds in iodine deficient children. DESIGN AND METHODS: 70, 70 and 72 schoolchildren of an area of severe iodine deficiency were studied before intervention (1989), 3 yr after injection of 480 mg iodized oil (1992) and 7 yr after consumption of iodized salt (1999), respectively. Goiter was graded and serum T4, T3, TSH and thyroglobulin concentrations and urinary iodine levels were measured. Audiometry was performed with a pure tone audiometer. RESULTS: There was significant decrease in the prevalence and severity of goiter and serum TSH and thyroglobulin concentrations, and significant rise in serum T4 in 1992 and 1999, as compared to 1989. Before iodine supplementation, hearing was abnormal in 44% of schoolchildren, mean hearing threshold was 15.8 +/- 5.9 and in all children was >10 dB. Mean hearing threshold decreased to 10.2 +/- 4.6 and 10.0 +/ 5.9, 3 and 10 yr after intervention (p < 0.001). Forty seven and 62% of children had thresholds < 10 dB in 1992 and 1999, respectively. Hearing thresholds > 15 dB were detected in 46, 11 and 10% of schoolchildren in 1989, 1992 and 1999, respectively (p < 0.001). CONCLUSION: Continuous iodine supplementation permanantly improves the auditory thresholds of iodine deficient children. PMID- 16218042 TI - A case-controlled study on the quality of life in a cohort of patients with history of differentiated thyroid carcinoma. AB - Although quality of life (QoL) has become an important aspect of cancer rehabilitation, psychometric studies on thyroid cancer patients are rare. We performed a case-controlled study on QoL in patients with differentiated thyroid carcinoma (DTC). QoL was evaluated in 61 patients with a history of DTC diagnosed from < 1 to 23 yr earlier. An undetectable thyroglobulin (Tg) level after recombinant human TSH (rhTSH) testing was considered the best predictor of cure. QoL was evaluated by means of a general psychiatric interview, the self-rating Kellner Symptoms Questionnaire (KSQ) and the Hamilton Depression Scale (HDS). QoL was also evaluated in a control group of subjects on L-T4 therapy with a non toxic multinodular goiter diagnosed from < 1 to 25 yr earlier. DTC and control subjects were similar in age, male-female distribution and concomitant psychiatric therapies. Per-week dosage of L-T4 was higher in DTC patients than in controls (p < 0.01). In neither group of subjects was there any correlation between current TSH levels or interval from diagnosis and KSQ or HDS scores. Only in DTC patients was there a positive correlation between age and KSQ (p < 0.05) or HDS (p < 0.01) scores. There was a significant difference in overall KSQ scores between DTC (33.4 +/- 2.1) and control (24.5 +/- 1.9; p < 0.01) subjects. The subscales of KSQ showed a significant inter-group difference. HDS scores were higher in DTC subjects (35.8 +/- 1.0) than in controls (30.0 +/- 1.1; p < 0.01). HDS score was significantly (p = 0.02) higher in female than in male DTC patients. In patients with papillary carcinoma there was a positive correlation between the MACIS (metastases, age, completeness, invasiveness, size) score and KSQ (p = 0.01) or HDS (p < 0.01) scores. After rhTSH testing, detectable Tg levels were found in 13% of DTC patients. In Tg-positive patients, KSQ and HDS scores were not different from those of Tg-negative patients. After an 8-14 month period, a significant decrease in the KSQ scale somatization (p = 0.02) was found in a sub-set of 31 DTC patients. In conclusion, even in the age of rhTSH testing, DTC patients suffer an impairment of their QoL, as noted when short-term L-T4 withdrawal was the gold standard. Longitudinal evaluation seems to indicate a slight improvement in QoL when safe rhTSH testing is extensively used in the management of the disease. PMID- 16218043 TI - Adiponectin values are unchanged during pregnancy in rats. AB - Adiponectin is believed to be a key factor in determining insulin sensitivity. In turn, insulin sensitivity is known to change from an enhanced state in early pregnancy to a reduced one in late pregnancy. A role for adiponectin in these changes has been proposed for mice but questioned for humans. We addressed this issue in rats by measuring adiponectin expression in both visceral and subcutaneous white adipose tissue, together with tissue content and release of the hormone in non-pregnant and in pregnant rats by days 8, 15 and 19 of pregnancy. Plasma concentration was also determined. No differences were found in any of the parameters measured between non-pregnant and pregnant rats at any time of pregnancy despite changes in white adipose tissue mass and insulin sensitivity. Adiponectin was also detected in cerebrospinal fluid at a concentration 1,000 times lower than in plasma, but again no differences were found between non-pregnant and pregnant animals. It is concluded that adiponectin does not play any role in regulating changes in insulin sensitivity during pregnancy in rats. PMID- 16218044 TI - Arterial stiffness in type 1 diabetes mellitus is aggravated by autoimmune thyroid disease. AB - OBJECTIVE: The aim of our study was to measure arterial stiffness in patients with Type 1 diabetes mellitus, its contributing factors and its relation to macrovascular arterial changes. MATERIALS AND METHODS: Thirty-one female Type 1 diabetic patients were studied; 11 had concomitant autoimmune thyroid disease although euthyroid during the study period. Stiffness was studied using applanation tonometry and pulse wave analysis for evaluation of systolic arterial pressure augmentation secondary to arterial stiffening and early wave reflection. Results were compared to 24 healthy individuals. In all patients, endothelium related flow-mediated dilation (FMD) of the brachial artery and intima-media thickness (IMT) of the carotid artery were measured. RESULTS: Augmentation pressure (AP) and augmentation index (AI) were higher in Type 1 diabetic patients suggesting stiffer arteries compared to controls (AP: 5.8 +/- 3.6 vs 2.8 +/- 2.2 mmHg, p < 0.001; and AI:18.3 +/- 9 vs 11.1 +/- 8.8%, p = 0.004). The subgroup of diabetic patients with autoimmune thyroid disease presented stiffer arteries than those without (AP: 6.5 +/- 2.9 vs 5.5 +/- 3.9 mmHg, p < 0.05; and AI: 21.3 +/- 5.4 vs 16.7 +/- 10.3%, p < 0.05), though the two groups did not differ statistically by means of age, disease duration, hemoglobin A1c (HbA1c), lipid levels, FMD and IMT. In multiple regression analysis, variables independently associated to AI in the diabetes group were: age (p = 0.028), IMT of the carotid artery bifurcation (p = 0.045), disease duration (p = 0.031) and autoimmune thyroid disease (p = 0.015). No correlation was observed between AI and metabolic control, blood pressure, microalbuminuria, presence of retinopathy and endothelial function (FMD). CONCLUSIONS: Women with Type 1 diabetes have increased arterial stiffness, which indicates macroangiopathy. An independent correlation between these indices and carotid IMT was observed. Concomitant autoimmune thyroid disease seems to aggravate arterial compliance in these patients, a finding that merits further investigation. PMID- 16218045 TI - Effects of raloxifene on body fat distribution and lipid profile in healthy post menopausal women. AB - The aim of our prospective, randomised, controlled and open-label clinical study was to evaluate in healthy post-menopausal women the effects of raloxifene (RLX) on body fat distribution and lipids, and the correlations between these parameters. The fat distribution, by dual energy X-ray absorptiometry, and lipids were evaluated at baseline and after 1 yr in 50 post-menopausal women: 25 were treated with RLX 60 mg/die, while 25 served as control group (CG). After 1 yr, we observed in RLX-users a slight reduction of fat mass in trunk and central region and an increase in legs and, in relation to CG, significantly lower values of adiposity in trunk and abdominal region (p < 0.05). At the same time, HDL cholesterol (HDL-C) and apolipoprotein A1 (ApoA1) were significantly increased in relation to baseline values and CG (p < 0.05) and apolipoprotein B (ApoB), total cholesterol/HDL-C, LDL cholesterol/ HDL-C, and ApoB/ApoA1 ratios significantly decreased compared to baseline values and CG (p < 0.05). No correlation was underlined among lipids and regional fat distribution. These results highlight the positive effect of RLX on lipids and suggest, for the first time, that RLX promotes the shift from android to gynoid fat distribution, and prevents the uptrend of abdominal adiposity and body weight compared with untreated women. PMID- 16218046 TI - Comparison of different regimens of glucocorticoid replacement therapy in patients with hypoadrenalism. AB - Since the optimal glucocorticoid replacement needs to avoid over and under treatment, the adequacy of different daily cortisone acetate (CA) doses was assessed in 34 patients with primary and central hypoadrenalism. The conventional twice CA 37.5 mg/day dose was administered to all patients (A regimen: 25 mg at 07:00 h, 12.5 mg at 15:00 h), while in 2 subgroups of 12 patients the dose was shifted on 2 thrice daily regimens (B: 25 mg at 07:00, 6.25 mg at 12: 00, 6.25 mg at 17:00; C: 12.5 mg, 12.5 mg, 12.5 mg). In other 12 patients the conventional dose was reduced to a thrice 25 mg/day administration (D regimen: 12.5 mg, 6.25 mg, 6.25 mg). In all patients, urinary free cortisol (UFC) excretion and cortisol day curves were evaluated. During the CA 37.5 mg administration, nadir cortisol levels were significantly higher with the thrice daily regimens (143 +/- 31 on B and 151 +/- 34 nmol/l on C) than with the conventional twice (85 +/- 16 nmol/l). Moreover, UFC, morning cortisol levels and mean cortisol day curves were similar in each group. Finally, during D regimen nadir cortisol levels were higher than in A and similar to B and C regimens. No difference in UFC and in cortisol day curves by reducing the CA dose was found. In conclusion, the thrice daily cortisone regimens, in which more physiological cortisol levels are achieved, perform better as replacement therapy. The administration of 25 mg/day CA confirms that replacement therapy is more adequate with a lower dose, particularly in patients with central hypoadrenalism. PMID- 16218047 TI - Role of intron 5 C/T polymorphism of the calcium sensing receptor gene in the regulation of the serum FSH and LH in post-menopausal women. AB - OBJECTIVE: High extracellular calcium concentration (Cao(2+)) acts to inhibit calcium sensing receptor (CaR) signalling on cellular surfaces in parathyroid glands. This receptor is, however, also expressed on the membranes of some non calciotropic endocrine cells, including pituitary-derived cells. The aim of our study was to analyse relationships between the CaR gene and the circulating FSH and LH in normal post-menopausal women. METHODS: A total of 95 untreated euparathyroid post-menopausal women were investigated in the study. The serum FSH and LH levels were evaluated in relationship to allele combinations of the CaR gene (C/T polymorphism in the intron 5 and A986S polymorphism in exon 7), using an analysis of co-variance (ANCOVA) model. RESULTS: Distribution of TT, TC and CC allele combinations (intron 5 C/T polymorphism) was 51, 43 and 6 %, respectively. Higher serum FSH and LH levels were found in carriers of C allele than in women without this allele (p < 0.002 and p < 0.03, respectively). No correlations were found between A986S polymorphism and serum FSH and LH levels. CONCLUSIONS: Serum FSH and LH levels are associated with intron 5 C/T (but not A986S) polymorphism of the CaR gene in untreated post-menopausal women. The physiological role of the CaR gene in the regulation of the gonadotropic function needs to be further investigated. PMID- 16218048 TI - Heterophile antibodies may cause falsely lowered serum cortisol values. AB - OBJECTIVE: We describe a case where interference by heterophile antibodies in a cortisol immunoassay led to a falsely lowered serum cortisol level. CASE DATA: As part of a diagnostic workup for fatigue, a 42-yr-old woman was found to have a low 9.00 h serum cortisol and an abnormal Synacthen stimulation test consistent with adrenal insufficiency. On insulin hypoglycemia testing, markedly discrepant cortisol levels were obtained from paired samples tested by two different immunoassays. Further tests, including plasma ACTH and overnight metyrapone test, confirmed normal hypothalamic-pituitary-adrenal axis function. Negative interference on the initial immunoassay from a heterophile antibody in the patient's serum was detected, thereby explaining the abnormal results. CONCLUSIONS: This report demonstrates that potentially significant consequences can arise from the failure to consider heterophile antibody interference. Clinicians who interpret test results need to be aware of the potential for interference in immunoassays by heterophile antibodies. PMID- 16218049 TI - Multiple intracranial calcifications and spinal compressions: rare complications of type la pseudohypoparathyroidism. AB - Type la pseudohypoparathyroidism (PHP la) is an unusual inherited disease. PHP la often causes extraskeletal calcifications and even soft tissue ossifications. Patients may present neurologic symptoms and signs related to hypocalcemia and hyperphosphatemia. We report here a 38-yr-old woman with PHP la who had two uncommon neurologic complications. One was involuntary movements related to basal ganglia calcification, and the other was myelopathy owing to ossifications of the posterior longitudinal ligament and multiple herniated intervertebral disks. Aggressive body weight control and corrections of hypocalcemia, hyperphosphatemia, and elevated PTH may be important to prevent these unusual neurologic complications. Regular and careful neurologic examinations should be performed for early diagnosis and treatments of these spinal lesions. PMID- 16218050 TI - A naturally occurring deletion in the SRY promoter region affecting the Sp1 binding site is associated with sex reversal. AB - Male to female sex reversal results from failure of testis development. Mutations in the SRY gene or in other genes involved in the sexual differentiation pathway are considered to cause XY gonadal dysgenesis. The majority of the mutations in the SRY described so far are located within the SRY coding region, mainly in the HMG-box conserved domain. Comparison of 5' flanking SRY gene sequences among different species indicated the presence of several putative conserved consensus sequences for different transcription regulators. In this study, we investigated a 360 bp sequence encompassing the SRY putative core promoter, in 17 patients with variable degrees of 46,XY sex reversal, which have been previously shown not to bear mutations in the SRYcoding region. Sequencing analysis of the SRYpromoter in one patient with complete XY gonadal dysgenesis revealed a three base pair deletion in one of the Sp1 binding sites. The deletion abolished Sp1 binding in vitro. This is the first report on a naturally occurring mutation affecting the Sp1 regulatory element in the SRY promoter region, which is associated with sex reversal. Additionally, upon familial investigation the father, who had 18 genital surgeries due to severe hypospadia without cryptorchidism, was found to bear the same deletion and several relatives were referred to have sexual ambiguity. PMID- 16218051 TI - Endocrine and neuropsychological assessment in a child with a novel mutation of thyroid hormone receptor: response to 12-month triiodothyroacetic acid (TRIAC) therapy. AB - OBJECTIVE: To determine the endocrine, neuropsychological and genetic features of a child with resistance to thyroid hormone (RTH), and his response to long-term triiodothyroacetic acid (TRIAC) therapy. METHODS: Growth, thyroid function, and neuropsychology were assessed at baseline and during 12-month TRIAC therapy. Genetic analysis was performed by PCR and denaturing high performance liquid chromatography. RESULTS: The main clinical finding was the attention deficit hyperactivity disorder (ADHD). A novel mutation in exon 10 (phenylalanine to isoleucine in codon 455) was found. Long-term TRIAC therapy was effective in the management of the endocrine and neuropsychological manifestations of the syndrome. CONCLUSIONS: ADHD was the only phenotypic manifestation of this novel mutation of thyroid hormone (TH) receptor. TRIAC is an effective and safe drug in the long-term treatment of children with RTH. PMID- 16218052 TI - Lack of TSH inhibition by exogenous L-T4 in a patient with follicular thyroid carcinoma. AB - We report a case of a male patient thyroidectomized for follicular thyroid carcinoma and presenting extremely elevated serum thyrotropin levels under L-T4 suppressive therapy. Administration of L-T3 in increasing amounts resulted in a significant decrease of serum TSH levels. The nature of the possible molecular defects underlying this unusual condition and pitfalls arising from the failure of L-T4 therapy to inhibit TSH secretion in a patient in post-surgical follow-up for follicular carcinoma are discussed. PMID- 16218053 TI - Andropause: need for concrete guidelines until more evidence becomes available. PMID- 16218055 TI - [Reflections on a new world]. PMID- 16218054 TI - A bronze statue of the tallest acromegalic giant. PMID- 16218056 TI - [Co-therapy with four-footed animals]. PMID- 16218057 TI - [Profound nursing knowledge makes deeper respect possible]. PMID- 16218058 TI - [Ambulatory psychiatric nursing. Surgery of an abscess]. PMID- 16218059 TI - [Politician as a help to nurses in the front]. PMID- 16218061 TI - [Mass nursing]. PMID- 16218060 TI - [No invasion from the East]. PMID- 16218062 TI - [Unrecognized understanding]. PMID- 16218063 TI - [Women's hearts beat differently from men's]. PMID- 16218064 TI - [Small joys happen]. PMID- 16218065 TI - [Vacations adapted to the needs of the child]. PMID- 16218066 TI - [Limits of autonomy]. PMID- 16218067 TI - [A unique offer for the Italian Swiss]. PMID- 16218068 TI - [Victory against discrimination]. PMID- 16218069 TI - ["Is it you, Pierrot?"]. PMID- 16218070 TI - [The sincerity of household pets]. PMID- 16218071 TI - Caveat emptor--because you get what you ask for. PMID- 16218072 TI - The Kleine-Levin syndrome: a paramedian thalamic dysfunction? PMID- 16218074 TI - RBD--an emerging clue to neurodegenerative disorders. PMID- 16218073 TI - Clinical and research implications of a validated polysomnographic scoring method for REM sleep behavior disorder. PMID- 16218075 TI - Influence of tetrodotoxin inactivation of the central nucleus of the amygdala on sleep and arousal. AB - STUDY OBJECTIVES: To determine the effects of temporary functional inactivation of the central nucleus of the amygdala on sleep and on activity in an arousing environment, an open field. DESIGN: Rats were implanted with electrodes for recording the electroencephalogram (EEG) and electromyogram (EMG), and with guide cannulae aimed into CNA. Sleep was recorded for 22 h (10 h light, 12 h dark) following microinjections of tetrodotoxin (TTX: 5.0 ng/0.2 microl given unilaterally [TTXUH] or bilaterally [TTXBH], and 2.5 ng/0.1 microl given bilaterally [TTXBL]) or saline (SAL) alone on separate days. Activity during 1 h in an OF was recorded after microinjections of TTXBH and SAL. SETTING: NA. PATIENTS OR PARTICIPANTS: Three-month-old Wistar rats (n=12). INTERVENTIONS: Functional inactivation of the central nucleus of the amygdala with TTX. MEASUREMENTS AND RESULTS: Compared to SAL, all TTX microinjections significantly shortened NREM latency, but did not alter total NREM during either light or dark periods. During the light period, TTXBH significantly reduced total REM and REM episode number, and TTXBL decreased REM episode number. All TTX microinjections increased EEG slow wave activity (0.5-4 Hz, SWA) during wakefulness, NREM and REM. Activity in OF was decreased after TTXBH compared to SAL. CONCLUSIONS: Functional lesions of the amygdala, including the central nucleus of the amygdala, decreased REM sleep and reduced arousal, as indicated by shortened NREM latency and decreased activity in an arousing environment. These findings suggest that the amygdala plays a broad role in modulating spontaneous sleep and wakefulness and in modulating responsiveness in arousing situations. PMID- 16218076 TI - Short-term homeostasis of REM sleep throughout a 12:12 light:dark schedule in the rat. AB - STUDY OBJECTIVES: Intervals extending from the end of a rapid eye movement (REM) sleep episode until the triggering of the next tend to be longer when they follow a longer REM sleep episode. A short-term REM sleep homeostatic process has been hypothesized to explain this effect. The present study assessed and modeled the REM sleep episode-interval relationship and compared its expression at different phases of a 12:12 light: dark schedule. DESIGN: Chronically implanted rats were continuously recorded for 3 consecutive days. Automated state scoring in 15 second epochs determined lengths of REM sleep episodes and intervals and non rapid eye movement sleep and wakefulness content of intervals. SETTING: Individual sound-attenuated temperature-regulated boxes. PARTICIPANTS: 16 Sprague Dawley rats. INTERVENTIONS: Scheduled 12:12 light:dark cycle. MEASUREMENTS AND RESULTS: The effect of REM sleep episode length is evidenced by a rising trend in the means and robust means of intervals and non-rapid eye movement content that follow REM sleep episodes of a given length. The relationship of robust means of intervals and REM sleep episode length was best fitted by a Gompertz sigmoid function. The parameters of the Gompertz equation were modulated throughout the 24 hours, presenting the highest amplitude and earliest rise in hours 1 to 4 after lights on and the lowest amplitude at the start of lights off. The modulation was also evident when only intervals with less than 3 minutes of wakefulness were considered. CONCLUSIONS: Short-term REM sleep homeostasis is modulated throughout the 24 hours under a 12:12 light:dark regime. Its assessment may provide a useful measure of REM sleep propensity, regulation, and recurrence. PMID- 16218077 TI - Clinical analyses of sighted patients with non-24-hour sleep-wake syndrome: a study of 57 consecutively diagnosed cases. AB - STUDY OBJECTIVES: The objective of this study was to clarify the clinical features of sighted patients with non-24-hour sleep-wake syndrome. DESIGN: Clinical analyses of consecutive patients suffering from non-24-hour sleep-wake syndrome. SETTING: The sleep disorders clinic at Kohnodai Hospital, National Center of Neurology and Psychiatry, Japan. PATIENTS: Fifty-seven patients who were diagnosed consecutively as having non-24-hour sleep-wake syndrome between 1991 and 2001 were included in the study. MEASUREMENTS AND RESULTS: The clinical features and sleep characteristics of the patients were analyzed. A semistructured psychiatric interview that included the criteria for Axis I or II disorders of Diagnostic and Statistical Manual of Mental Disorders, Third Edition Revised was conducted, and relationships between psychiatric problems and non-24 hour sleep-wake syndrome were analyzed. The patient cohort included 41 (72%) men and 16 (28%) women. The onset of non-24-hour sleep-wake syndrome had occurred during the teenage years in 63% of the cohort, and the mean ( +/-SD) period of the sleep-wake cycle was 24.9 +/- 0.4 hours (range 24.4-26.5 hours). The mean sleep length of the patients was 9.3 +/- 1.3 hours, and 44% of them had a sleep length of between 9 and 10 hours. Psychiatric disorders had preceded the onset of non-24-hour sleep-wake syndrome in 16 patients (28%); of the remaining 41 patients, 14 (34%) developed major depression after the onset of non-24-hour sleep-wake syndrome. CONCLUSIONS: These results represent the first detailed clinical review of a relatively large number of sighted patients with non-24-hour sleep-wake syndrome. PMID- 16218078 TI - SPECT findings in the Kleine-Levin syndrome. AB - STUDY OBJECTIVES: The Kleine-Levin Syndrome, is a rare disorder with onset during teenage years, but little is known on etiopathogenesis. Seven subjects with Kleine-Levin Syndrome accumulated over time had systematic SPECT studies during (n=5) and out (n=7) of the symptomatic period. SUBJECTS: Seven boys with symptom onset between 11 and 17 years of age and at least 2 episodes per year were followed for a mean of 6 years. METHODS: Electroencephalogram awake-asleep, computed tomography scan, and magnetic resonance imaging studies were performed before Tc-99m ECD single photon emission tomography (SPECT) obtained during day 4 or 5 (n=5) and at least 1 month away from the symptomatic period (n=7). RESULTS: All imaging tests except SPECT were normal. Hypoperfusion of both thalami were seen during the symptomatic period that completely disappeared during the asymptomatic period. Hypoperfusion in other regions were also noted in some, but not all subjects. They persisted during the asymptomatic period in 2 cases over the temporal lobe (2/7 cases), frontal lobe (1/7 cases), and basal ganglia (1/7 cases). The largest amount of persistent hypoperfusion was seen in the subject with longest clinical evolution. CONCLUSION: Hypoperfusion of the thalamus is a consistent finding during the symptomatic period, but perfusion abnormalities may persist even during the asymptomatic period. The longer the duration of the syndrome, the more extended the hypoperfusion regions during the asymptomatic period. PMID- 16218079 TI - Does insomnia kill? AB - STUDY OBJECTIVES: We investigated the prevalence and hazard ratios for insomnia complaints in a large cohort of middle-aged men and women. DESIGN: The Atherosclerosis Risk in Communities Study is a prospective study of cardiovascular disease. Using multivariate regression analysis, we predicted the likelihood of endorsing the insomnia complaints by age, sex, alcohol intake, smoking, diabetes, heart disease, menopausal status, use of hypnotics, hypertension, depressive symptoms, education level, body mass index, respiratory symptoms, and pulmonary function status. We predicted the hazard ratios (HR) of death at 6.3 +/- 1.1 year by endorsement of insomnia complaints and by hypnotic use controlling for covariates. SETTING: North American communities. PARTICIPANTS: 13563 participants aged 45 to 69 years at baseline INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The prevalence of insomnia complaints in this cohort was 23%. Predictors of insomnia complaints were female sex (odds ratio [OR] 0.56, 95% confidence interval [CI] 0.45-0.70 for men), annual family income below 50,000 dollars (OR 1.23, CI 1.09-1.40), age 40 to 49 years (OR 1.29, CI 1.11-1.50), depressive symptoms (OR 5.05, CI 4.60-5.55), heart disease (OR 1.89, CI 1.67-2.14), severe airflow obstruction (OR 1.61, CI 1.17-2.22), pulmonary symptoms (OR 1.71, CI 1.5-1.95), and restrictive lung disease (OR 1.27, CI 1.10 1.47). After controlling for covariates, insomnia complaints were not associated with an increased risk for death (OR 1.01, CI 0.85-1.21), nor was the use of hypnotics (OR 1.38, CI 0.90-2.13). CONCLUSIONS: In this cohort, the prevalence of insomnia complaints was 23%. After controlling for confounders, neither insomnia complaints nor hypnotic use predicted increased mortality over 6.3 years. PMID- 16218080 TI - The experience of insomnia among older women. AB - STUDY OBJECTIVES: To measure sleeping difficulty and sleep quality among older women, explore experience and attitudes towards sleep, and test for negative association between difficulty sleeping and health-related quality of life. DESIGN: Four-year longitudinal study. SETTING: Women were participants in the Australian Longitudinal Study on Women's Health. PARTICIPANTS: Women were sampled according to use of sleeping medication and classified into 4 groups: sleeping badly and using sleeping medications; not sleeping badly, but using sleeping medications; sleeping badly, not using sleeping medications; not sleeping badly, not using sleeping medications. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Sleeping difficulty and sleeping-medication use were measured at Survey 1, Survey 2 (3 years later), and Survey 3 (4 years later). Survey 3 included: Nottingham Health Profile Sleep Subscale, Pittsburgh Sleep Quality Index, Epworth Sleepiness Scale, Geriatric Depression Scale, Duke Social Support Index, Medical Outcomes Study Short-Form 36-item Health Survey, and a 21-item life events scale. Survey 3 was returned by 1011 women (84%). Sleeping problems were negatively associated with SF-36 subscale scores. Most associations remained significant after comorbid conditions, Geriatric Depression Scale, life events scores, and medication use were added to models. Most women with sleeping problems (72%) sought help from a doctor, and 54% used prescribed sleeping medications in the past month. CONCLUSIONS: Sleeping difficulty is a serious symptom for older women and is associated with poorer quality of life. Some of this effect can be explained by comorbidities, depression scores, life events, and use of sleeping medications. PMID- 16218081 TI - Normative sleep data, cognitive function and daily living activities in older adults in the community. AB - STUDY OBJECTIVES: To present normative data of sleep-wake characteristics and to examine risk factors associated with extreme values (i.e., in the 5 lower and upper percentiles of the distribution) in older adults. DESIGN: Cross-sectional telephone survey SETTING: The metropolitan area of Paris, France. PARTICIPANTS: A total of 7010 randomly selected households were contacted. Among them, 1264 households included at least 1 resident 60 years of age or older; 1026 subjects agreed to participate (participation rate: 80.9%). INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Subjects were interviewed with the Sleep-EVAL System about their sleeping habits and sleep and psychiatric disorders. In addition, the system administered to all the participants the Psychological General Well-Being Schedule, the Cognitive Difficulties Scale (Mac Nair-R), and an independent living scale. The median nighttime sleep duration was 7 hours without significant difference between the age groups. Factors positively associated with the 5 percentile (4 hours 30 minutes or less) of nighttime sleep duration were obesity, poor health, insomnia, and insomnia accompanied by daytime sleepiness and cognitive impairment. At the other extremity (95th percentile), long sleep (9 hours 30 minutes or more) was associated with organic disease, lack of physical exercise, and lower education. A daytime sleep duration of 1 hour or more (95th percentile) was associated with being a man, cognitive impairment, high blood pressure, obesity, and insomnia. Long sleep latency (95th percentile at 80 minutes) was associated with anxiety, lower education, poor health, insomnia without excessive daytime sleepiness, and obstructive sleep apnea syndrome. Obesity and loss of autonomy in activities of daily living was associated with both early (9 PM or earlier) and late bedtime (1 AM or later) and early (< or = 5 AM) and late (> or = 9 AM) wake-up time. CONCLUSIONS: This study illustrates the usefulness of normal distributions of sleep parameters in the general population to calculate different risk factors associated with extreme values of the normal distribution. PMID- 16218082 TI - Effect of using the flow or the volume signals on the measurement of nonapneic respiratory events. AB - STUDY OBJECTIVES: To assess whether the measurement of breathing reduction during obstructive sleep events depends on using the flow or the volume signals recorded with a pneumotachograph. DESIGN: Prospective observational study. SETTING: Sleep laboratory in a University Hospital. PATIENTS OR PARTICIPANTS: Data from 10 male patients with sleep apnea (54 +/- 11 years, apnea-hypopnea index: 43 +/- 21 events/hour, body mass index: 30 +/- 2 kg/m2). INTERVENTIONS: Slow modification of continuous positive airway pressure was performed during full-polysomnography continuous positive airway pressure titration. MEASUREMENTS AND RESULTS: Air flow was measured by a pneumotachograph, and volume was computed by numerical integration. Obstructive events of different magnitude were selected. In 500 breathing cycles analyzed, the reduction in tidal volume was greater than the reduction in the flow amplitude: mean difference of 0.091 (i.e., 9.1% amplitude) and limits of agreement of 0.095 and -0.277 (i.e., 9.5% and -27.7% amplitude). In 14% of the cycles, the reduction in flow was < 50%, whereas the reduction in volume was > 50%, resulting in discordant event classification. CONCLUSIONS: The quantification of breathing reduction depends on whether the flow or the volume signal is used to assess breathing during sleep. PMID- 16218083 TI - Validation of a polysomnographic score for REM sleep behavior disorder. AB - STUDY OBJECTIVES: Rapid eye movement (REM) sleep behavior disorder (RBD) was described more than 2 decades ago, but only 1 report on 5 patients and 5 normal subjects has tested the effectiveness of a method by which relevant polysomnographic findings can be quantified. We sought to validate this method in a larger sample of patients and control subjects. DESIGN: Cross-sectional. SETTING: Academic hospital. INTERVENTIONS: A clinician interviewed 17 patients at risk for RBD secondary to neurodegenerative disorders and 6 controls to assess whether RBD was present by history. Bed partners completed a questionnaire that quantified RBD symptom severity. From 2 consecutive nocturnal studies in each patient, 2 different polysomnographic RBD scores were generated: the percentage of 30-second REM epochs with at least 15 seconds of tonically maintained electromyographic activity, and the percentage of 3-second REM mini-epochs that contained phasic electromyographic bursts. MEASUREMENTS AND RESULTS: The tonic and phasic measures, combined together, were higher in patients with clinical determinations of probable or possible RBD (n=9) than in patients judged unlikely to have RBD (n=4, P = .023). The overall polysomnographic measure correlated with the symptom scores (rho = 0.42, P = .048). Specific polysomnographic RBD measures on night 1 correlated highly with those on night 2 (rho > 0.70, P < .0001). CONCLUSIONS: This quantitative method to assess the severity of RBD polysomnographic features appears to be both valid and reliable in patients at risk for RBD because of neurodegenerative disorders. PMID- 16218085 TI - Restless legs syndrome and attention-deficit/hyperactivity disorder: a review of the literature. AB - STUDY OBJECTIVE: To review evidence on the association between restless legs syndrome (RLS) and attention-deficit/hyperactivity disorder (ADHD), to discuss the hypothetical mechanisms underlying this association, and to consider the potential interest for common pharmacologic treatments of RLS and ADHD when co occurring. METHOD: A PubMed search. RESULTS: In clinical samples, up to 44% of subjects with ADHD have been found to have RLS or RLS symptoms, and up to 26% of subjects with RLS have been found to have ADHD or ADHD symptoms. Several mechanisms may explain this association. Sleep disruption associated with RLS might lead to inattentiveness, moodiness, and paradoxical overactivity. Diurnal manifestations of RLS, such as restlessness and inattention, might mimic ADHD symptoms. Alternatively, RLS might be comorbid with idiopathic ADHD. Subjects with RLS and a subset of subjects with ADHD might share a common dopamine dysfunction. Limited evidence suggests that some dopaminergic agents, such as levodopa/carbidopa, pergolide, and ropinirole, may be effective in children with RLS associated with ADHD symptoms. CONCLUSIONS: Although still limited, evidence from clinical studies demonstrates an association between RLS and ADHD or ADHD symptoms. Further clinical studies using standard criteria and procedures are needed to better estimate the degree of association. Epidemiologic studies are required to assess the relationship between ADHD and RLS symptoms in nonclinical samples. Further investigations should address the mechanisms underlying the relationship between RLS and ADHD. Several dopaminergic agents seem to be promising treatment for RLS associated with ADHD symptoms. To date, however, the absence of randomized and blinded controlled studies does not allow evidence based recommendations. PMID- 16218084 TI - Computer-assisted detection of nocturnal leg motor activity in patients with restless legs syndrome and periodic leg movements during sleep. AB - STUDY OBJECTIVES: To assess the performance of a new method for automatic detection of periodic leg movements during sleep. METHODS: Leg movements during sleep were visually detected in the tibialis anterior muscles recordings of 15 patients with restless legs syndrome and 15 normal controls. Leg movements were detected automatically by means of a new computer method with which electromyogram signals are first digitally band-pass filtered and then rectified; subsequently, the detection of leg movements is performed by using 2 thresholds: one for the starting point and another to detect the end point of each leg movement. Sensitivity and false-positive rate were obtained; the American Sleep Disorders Association parameters were also computed, and the results analyzed by means of the Kendall W coefficient, the linear correlation coefficient and the Bland-Altman plots. SETTING: N/A. PARTICIPANTS: Fifteen patients with restless legs syndrome and periodic leg movements and 15 controls. MEASUREMENTS AND RESULTS: High values of the Kendall W coefficient of concordance between automatic and visual analysis were found with values close to 1 and the linear correlation coefficient for leg movements index and total leg movements index was > 0.950 (p < .000001). The Bland-Altman plots provided the limits of agreement between visual and computer detection, which were -9.01 and +9.89 for the periodic leg movement index. None of the normal controls was found to have periodic leg movement indexes >5 after automatic analysis. CONCLUSIONS: Our method can be applied to the clinical evaluation of periodic leg movements during sleep, with some caution in patients with a low periodic leg movement indexes. Large-scale research application is possible and can be considered as reliable. PMID- 16218086 TI - A pharmacotherapy for obstructive sleep apnea. PMID- 16218087 TI - Statistical artifact in the validation of actigraphy. PMID- 16218088 TI - How to assess endothelial function in patients with obstructive sleep apnea? PMID- 16218089 TI - [Individual developmental care based on the Newborn Individualized Developmental Care and Assessment Program (NIDCAP)]. AB - The number of preterm deliveries is increasing. Major advances in neonatology now enable even infants with a very low gestational age to survive. These infants have to spend months in the intensive care unit, while they have to deal with a lot of painful medical procedures, nursing care units, excessive noise and light, all differing from the normal uterine environment. The immature preterm infant nervous system is in a vulnerable phase of development. Early exposure to stress, or the mismatch of the preterm infant brain with the unphysiologic intensive care environment, may be linked to long-term developmental impairments reported in these children. The Newborn Individualized Developmental Care and Assessment Program (NIDCAP) is a systematic method of assessing the individual needs of preterm newborns. In an attempt to mimic uterine environment, care according to the NIDCAP principles is reducing infant distress and in the same way supporting the individual aibilities. Thus NIDCAP may have an effect on the preterm infant neurodevelopmental outcome. PMID- 16218090 TI - [Typical: the Bremen model of the family midwife and family pediatric nurse]. PMID- 16218092 TI - [Early diagnosis of speech development disorders]. PMID- 16218091 TI - [Reader's letter on the article "New pediatric illnesses--effect of universal living conditions on health and development" by H. Schlack, kks 6/200, p. 245]. PMID- 16218093 TI - [The mental status of children before an operation]. PMID- 16218094 TI - [Dry eyes. Tears absent only rarely]. PMID- 16218095 TI - [Interview with the autistic book author Axel Brauns]. PMID- 16218096 TI - [Back schools. Does the back have to go to school?]. PMID- 16218097 TI - [What is the glycemic index worth?]. PMID- 16218098 TI - [Probiotics in the pediatric and adolescent medicine -- health promotion or vogue?]. PMID- 16218099 TI - [Delegation of physician's activities to unqualified personnel is illegal and can have penal consequences]. PMID- 16218100 TI - [Principles of the delegation of injections, infusions and collection of blood to nursing personnel (or other nonphysician personnel)]. PMID- 16218101 TI - [Questions on mammography screening or breast cancer prevention?]. PMID- 16218102 TI - ["My child is different"]. PMID- 16218103 TI - [Nothing is impossible or the biggest fantasy in the world]. PMID- 16218105 TI - [Joint leaflet of logopedia specialists and pediatricians on child language development]. PMID- 16218104 TI - [Risks to life in legal accident insurance excluded]. PMID- 16218106 TI - [Vitamin C is nevertheless not a common cold killer]. PMID- 16218107 TI - [Fine dust alert not only on much travelled streets?]. PMID- 16218108 TI - [Reader's letter on the article "Cesarean section on demand -- the legend of easy delivery" by A. Mechkat, kks 6/2005, p.254]. PMID- 16218109 TI - Is the UK prepared for a new flu pandemic? PMID- 16218110 TI - Let's end the stigma of mental illness. PMID- 16218111 TI - Primary care pioneers. PMID- 16218112 TI - NT campaign backs nurses. PMID- 16218114 TI - Insomnia. PMID- 16218113 TI - Government initiatives to tackle the obesity epidemic. AB - This article highlights the prevalence of obesity in the UK and the associated health costs. It discusses the attempts the government is making to halt the growth of the 'obesogenic' society and summarises practical tips and interventions of use when advising individuals on obesity issues. PMID- 16218115 TI - Taking a swab. PMID- 16218116 TI - Administering drugs to patients with swallowing difficulties. AB - Dysphagia is common among older patients. Its impact can be profound can swallowing difficulties can not only undermine the quality of life and exacerbate malnutrition but also complicate medication administration. Tablet crushing and capsule opening are widespread, yet few health care professionals are aware of the pharmacological or legal aspects of these practices. This article examines some of the causes of dysphagia in older patients, its impact on medication management and practical ways for nurses to meet this challenge. PMID- 16218117 TI - Sumatriptan and zolmitriptan. PMID- 16218118 TI - Development of a guide for neurological observations. AB - Following publication by the Scottish Intercollegiate Guidelines Network (SIGN) of Early Management of Patients with a Head Injury, local practices were reviewed in a Glasgow trust. This led to both organisational and clinical changes. This article discusses how a training programme in performing neurological observations was developed to meet staff learning needs regarding head injury, and explains changes to the Glasgow Coma Scale (GCS) charts used in the trust. PMID- 16218119 TI - A nurse-led e-mail service for breast cancer information. AB - AIM: To discover why people access the web-based 'Ask the nurse' service, what type of information they require and their overall level of satisfaction. METHOD: A prospective sample of users was invited to respond to an e-mail questionnaire. RESULTS: The results show the service is being used not only by people seeking clarity before or after diagnosis and treatment, but is also used by many people as an alternative to visiting a health care professional (GP or other doctor or breast care nurse). Indeed, 36 per cent of people in this study did not appear to know who else to ask, while others found the service accessible because they found it easy and convenient. CONCLUSION: E-mail communication has advantages, but health care professionals need to be aware of potential problems before offering this type of communication to their patients. PMID- 16218120 TI - Remember those with special needs. PMID- 16218121 TI - Infection control in learning disability services. AB - Services for people with a learning disability are provided by many sectors of the health and social care economy. These include: social services, health services, voluntary organisations, charities, private care agencies and family carers. Care interventions can take place in a variety of settings, from the client's own home to day care, respite care, educational establishments, workshops, social clubs, luncheon clubs, shared housing and the acute services. PMID- 16218122 TI - Legionella bacteria and water systems in health care premises. AB - Legionnaires' disease (Legionellosis) is a bacterial pneumonia that acquired its name following an outbreak of pneumonia among army veterans attending an American Legion convention in Philadelphia in 1976. The previously undiscovered bacterium that caused this form of pneumonia was therefore named Legionella. There are more than 45 species of Legionella bacteria and these are natural inhabitants of water supplies throughout the world (Fliermans, 1981). About half of these have been associated with infection, although the most common cause of Legionnaires' disease is Legionella pneumophila, which is the causative strain in around 90 per cent of cases (Fields et al, 2002). PMID- 16218123 TI - Exposure to blood or body fluids: management for health care. AB - Health care workers are at risk of exposure to blood and body fluids from their patients through needlestick injuries or contamination of mucous membranes - a slash of blood in the eye, for example. Exposure to blood /body fluids is now the second biggest cause of occupational injury among NHS workers (UNISON, 2003). The first example of a case of occupational exposure of a health care worker to human immunodeficiency virus (HIV) and subsequent seroconversion following a needlestick injury was reported in the 1980s. This incident raised awareness of the risk that health care workers face when they are exposed to blood-borne viruses, in particular hepatitis B, hepatitis C and HIV. PMID- 16218124 TI - Mumps: current outbreaks and vaccination recommendations. AB - Mumps is a moderately infectious disease caused by a virus of the paramyxovirus group. The infection is spread by airborne droplets and possibly also by urine. The first symptoms occur after an incubation period of 15-24 days (median 19 days) (Richardson et al, 2001). After a prodromal (warning signs) period of several days with non-specific flu-like symptoms of headache and fever, the classic swelling of the parotid glands (salivary glands) develops. The swelling may be unilateral or, in 90 per cent of cases, bilateral. It is accompanied by abdominal pain and headache and usually resolves in 7-10 days. However, there may be other symptoms of mumps without parotid swelling. Almost one-third of mumps infections are subclinical or mild, with non-specific symptoms, and are never diagnosed as mumps (Gupta et al, 2005). PMID- 16218125 TI - The benefits of team training. PMID- 16218126 TI - Where no two days are the same. PMID- 16218127 TI - Ceramic corundum materials with modified surface as implantation materials. AB - Investigations on implants have proved that not only the kind of material structure, but also the surface character of grafts influences the local reaction of biomaterials. In recent years electrochemical reactions on implants surface leading to the so called zeta potential arouse interests. Applications of these properties has made us carry on experiments on use of biomaterials with active potential on their bio-compatibility. The aim of this work is evaluation of influence of electrical charge with zeta potentials character on the local reaction of bone tissue after implantation. Two kinds of ceramics were used in experiments: solid and solid-silane with active surface. Implantation experiments were made on 20 rabbits. Selections were carried of 12, 26, 36 and 54 weeks after implantation of the tested materials. After implantation of solid ceramics, not only bone tissue but also fibrous tissue was observed around the graft. But in case of solid silane ceramics, in all terms of tests, bone tissue tightly sticking to the implant was observed around the graft. Investigations of electrokinetic zeta potential carried out 36 and 54 weeks after implantation showed that the values of that potential originally introduced on the grafts surface were maintained on stable level. The carried out investigations confirmed that activation of implants surface through silaning with sol-gel method allows to introduce the planned zeta potential depending on the characteristic for a given kind of tissue. PMID- 16218128 TI - [Biophysical properties of membrane dressing made of bacterial cellulose]. AB - In the paper, review of papers devoted to biophysical properties of membrane dressing made of bacterial cellulose was done. These properties were determined on the basis of studies on osmotic and diffusive transport through pure (non modified) bacterial cellulose membrane form called Bio-Fill. The measures of these properties are values of membrane transport parameters resulted from Kedem Katchalsky's theory and interferograms of near-membrane regions made laser interferometric method. PMID- 16218129 TI - [Medical properties of the membrane dressing made of bacterial cellulose]. AB - Review of papers devoted to medical properties of membrane dressing made of bacterial cellulose was done. These properties were determined on the basis of studies on application of this membrane to venous leg ulcer healing. Moreover, quantitative method of valuation of wound healing process efficiency which lies in calculating efficiency coefficient was described. Value of this coefficient is directly proportional to ulcer healing speed and indirectly proportional to product of initial surface and healing time. PMID- 16218130 TI - [The evaluation of early and late results of using Codubix cranial prosthesis]. AB - The filling of the cranial defect is an essential problem in neurosurgery on account of the necessity of brain protection as well as for cosmetic reasons. The aim of this study was to evaluate the direct and distant outcome after polypropylene-polyester Codubix prosthesis implantation. This research was based on the analysis of 41 patients treated surgically in the Department of Neurosurgery at the Jagiellonian University in Cracow between 1995-2004. All patients had the Codubix prosthesis implanted. The implantation was performed either synchronously or after the surgery which caused the cranial defect. Directly after the surgery proper healing of the prosthesis could be observed in all patients. The exception was one patient who died directly after the surgery but the cause of his death was not connected with the prosthesis implantation. Seven patients (17,1%) had the temperature temporarily elevated. Three had haematoma collection in the subgaleal space, one (2,4%) developed meningitis which was successfully treated with antibiotics, one (2,4%) suffered from temporary circulation disturbances of the scalp. The late evaluation was performed with help of a questionnaire to which 30 patients (73,2%) responded. The answer ranged 73,2% (30 patients). In this group 16 patients (53,4%) described the cosmetic effect as very good, 10 patients (33%) as good and 4 (13,3%) as unsatisfactory. The commonest reason for the critical note was the depression of the plate which happened to 3 patients. The other 3 patients complained about inappropriate profile of the plate causing the asymmetry of the cranial vault. Seven patients felt temporary pain in the postoperative scar, in one case the pain was took as the friction between the prosthesis and bone margin. Properly performed implantation of accurate bone prosthesis Codubix is safe and usually brings a good cosmetic effect. Direct complications are transitional and occur in the small percent of cases. Distant examinations show generally good therapeutic results but implanted Codubix requires suitable shape and careful fixation. PMID- 16218131 TI - [Biomaterials and tissues material in the treatment of prosthetic grafts infections]. AB - An article is presented the treatment of vascular prosthetic grafts infections. An the graft infection treated by the replacement of infected prosthesis with autogenic venous material or with venous and arterial allograft harvested from brain-dead organ donors together with multiple organ procurement is presented. Autogenous material has an ability a better healing in infected tissues and used with absorbable sutures may lead to complete recovery from vascular graft infection - a severe and often lethal complication. An article is presented the treatment of vascular prosthetic grafts infections with the use of more resistant prostheses of infection - silver coated prosthesis, prosthesis with antibiotic and polytetrafluoroethylene prostheses. PMID- 16218132 TI - [Human resources and activities in the occupational medicine service of Poland in 2003]. AB - BACKGROUND: The presented paper shows the most essential data concerning the state of human resources and activities of occupational medicine services in Poland in 2003. MATERIALS AND METHODS: The reports (around 10 000) obtained from primary occupational medicine service units and regional occupational medicine centers (forms: MZ-35A and MZ-35) are the source of information about human resources and occupational medicine services activities. RESULTS: A majority of regional centers, due to their restructuring, have approached the legal and organizational model described in the Occupational Medicine Service Act. The assessment of activities of primary occupational medicine centres units, performed by regional occupational medicine centers, indicates the need for constant training of physicians authorized to perform prophylactic examinations of employees. CONCLUSIONS: The year 2003 did not witness any new significant changes in the structure of occupational medicine service. One can speak rather about the continuation of trends observed in previous years. PMID- 16218133 TI - [Exposure to pesticides among pregnant women working in agriculture]. AB - BACKGROUND: Pesticides have been used since the early days of modern agriculture. They are biologically active compounds that may pose a grave risk to health during or after their use. The available data, documenting pesticide exposures and related health effects in farmers, especially in pregnant women, are limited. MATERIALS AND METHODS: Women were informed that the purpose of the research was to gather information about the exposure to pesticides during pregnancy. They were asked to provide information about their work in the field, spraying or preparatory work (washing clothes after spraying, cleaning, spraying equipment, preparing mixture for spraying). We received information about all spraying in household during woman's pregnancy, trade names of pesticides, names of active ingredients, type of cultivation and its area. RESULTS: The survey of pregnant women working in agriculture provided us with knowledge of potential hazards resulting from their work during pregnancy, especially from their involvement in spraying or preparatory work and working in the field after spraying. Other data obtained informed us what kind of pesticides were used in agriculture. CONCLUSIONS: The results indicate the necessity for increasing awareness of potential exposure to pesticides and their harmful effect among workers, especially among pregnant women. There is also a need to estimate real exposure via biological monitoring. PMID- 16218134 TI - [Knowledge of the employees about biological agents in the work environment and the ways of health protection against occupational exposure to biological hazards. Part II]. AB - BACKGROUND: The effectiveness of health protection of workers occupationally exposed to biological agents is equally influenced by the workers' knowledge about the risks related to biological agents and the ways they behave at work. MATERIALS AND METHODS: In this paper, the authors continue to present the results of the study covering 150 workers employed in health services, forestry, and municipal services. They were interviewed on the basis of a specially designed questionnaire. RESULTS: The majority of the study population used personal protective devices in a proper way and followed the principles of occupational safety and hygiene. Only some of the workers informed occupational medicine physicians about their health problems, which they associated with occupational exposure to biological agents. CONCLUSION: The interviewed workers are actually able to sufficiently protect their health against biological hazards despite their generally low knowledge of risks related to occupational exposure to biological agents. PMID- 16218135 TI - [Exposure assessment to harmful agents in workplaces in sewage plant workers]. AB - BACKGROUND: The purpose of the study was to evaluate exposure to biological and chemical agents in a sewage treatment plant. MATERIALS AND METHODS: Sampling was carried out in the summer and wintertime at the morning workshift. Ninety-nine sewage workers taking part in the study were divided into four occupational subgroups: mechanical treatment, biological treatment, sewage sludge treatment, and operation control workers. Exposure to: H2S, SO2, Pb, Cd, Cr3+, Cr6+, endotoxins, (1 --> 3)-beta-D glucans, and microorganisms was evaluated with special identification of Gram-negative rods. RESULTS: The concentrations of dust containing heavy metals and concentrations of gases from all stations did not exceed MAC values. Concentrations of endotoxins ranged from 0.08 to 223 ng/m3, and glucans from 0.00 to 163 ng/m3. The highest concentrations were found among sewage sludge treatment workers, in the summertime (geometric mean value = 37 ng/m3). In the winter, concentrations were almost ten times lower. Over sixty percent of all results exceeded the proposed reference value for airborne endotoxins (10 ng/m3). Concentrations of airborne bacteria in the sewage plant were at low level (10(2)( cfu/m3), except the sludge lagoon and sludge concentration building, where the results exceeded the proposed reference value for mesophilic bacteria (10(5) cfu/m3) "Environmental" bacteria (Pseudomonas, Burkholderia, Shewanella) predominated in the samples. There were also found enterobacteria genus (Enterococcus, family Enterobacteriaceae)--good indicators of hygienic cleanliness of the air. CONCLUSIONS: The study proved that the exposure varied and depended on the stage of sewage treatment. The sewage sludge treatment process was characterized by the highest emission of bioaerosols. All microorganisms found in the sewage plant belong to the second occupational risk group, under the ordinance of the Ministry of Health. PMID- 16218136 TI - [Trait anxiety and type behavior pattern (A and B) as modifiers of immediate reaction towards violent behaviors]. AB - BACKGROUND: Experience of violence at the work place leads to serious consequences for both an employee and the organization. That is why prevention programs are necessary to reduce the risk of violence at work as well as possible adverse consequences of violent acts that cannot be predicted and prevented. Some research studies suggest that the range and severity of individual consequences of experiencing violence at work depend on the personality of victims, including their typical reactions to violent acts. In this paper, such personal characteristics as trait-anxiety, type A/B behavior pattern (TABP/TBBP) are discussed as potential modifiers of individual reaction towards the violence experienced at work. MATERIALS AND METHODS: A sample of 1163 nurses, representative of the Mazovian Voivodship was surveyed. The standardized "paper and pencil" questionnaires were employed in the survey. The relationship between direct reaction to violence and trait-anxiety and TABP/TBBP was analyzed. RESULTS: Tendency to express an aggressive reaction towards violent behavior of others was positively correlated with a high level of trait-anxiety and TABP and submissive reactions to violence with a high level of trait-anxiety and TBBP, whereas tendency to express assertive reaction when being attacked was related to a low level of trait-anxiety and TBBP. CONCLUSIONS: Statistically significant relationship between trait-anxiety and behavior pattern was observed. A high level of anxiety is correlated with ineffective coping with violent acts as submissive and aggressive reaction. The tendency to express these two kinds of behavior when facing violence is strengthen by behavior pattern. The tendency to aggressive reactions as a response to aggression of others is stronger when a high level of trait-anxiety is accompanied by TABP. The tendency to present submissive reactions as a response to aggression of others is stronger when a high level of anxiety and TBBP characterize the individual who faces a violent act. These individual characteristics are thought to be rather stable and difficult to change, but they should be taken into account in the process of designing the violence prevention programs for the organization. We believe that in our attempts to reduce the extent of the problem we should provide information on the role of individual characteristics in the process of coping with violence and recommend to employ positive effects of the repeated assertiveness training in the violence prevention programs as a useful tool for teaching people how to behave in the face of aggressiveness of others. PMID- 16218138 TI - [Assessment of respiratory health status in the population of middle school students]. AB - BACKGROUND: The aim of the study was to assess the prevalence of asthma and to identify the basic host and environmental risk factors for asthma and symptoms suggestive of asthma in Polish adolescents aged 15 years. MATERIALS AND METHODS: The study was performed as a cross-sectional questionnaire survey comprising population of 854 - 68.3% of all eligible students, attending schools located in Jaworzno. Parents or guardians filled the questionnaire, which contained questions on the occurrence of respiratory and allergic symptoms and diseases, family history of these pathologies, data on exposure to indoor factors and socioeconomic status of the family. Statistical analyses involved calculation of the prevalence of asthma and symptoms suggestive of asthma and identification of risk factors on the basis of multivariate logistic regression. RESULTS: Wheezing in the last year preceding the survey was reported in 44 (5.1%) adolescents, attacks of dyspnea with wheezing in 22 (2.6%), and asthma in 46 (5.4%) adolescents. The results of multivariate logistic regression revealed that the presence of atopy was the most important risk factor for asthma (odds ratio: OR - 7.8; p = 0.0001). Other statistically significant risk factors for asthma and symptoms suggestive of asthma were positive history of allergy in mothers (OR - 8.3; p = 0.04) and positive history of severe respiratory disease in early childhood (OR - 4.7; p = 0.007). CONCLUSIONS: The results of our study justify the need for activities aimed at proper job orientation in future for adolescents with the presence of atopy. PMID- 16218137 TI - [Efficacy of therapeutic exercises in low back pain surveyed in a group of nurses]. AB - BACKGROUND: The aim of the survey was to analyze the efficacy of therapeutic exercises in a selected group of nurses and to show the interest of nurses in kinesitherapy as a method for preventing back pain complaints. MATERIALS AND METHODS: A questionnaire survey covered a group of 102 active nurses, being at the same time part-time students of the Poznan Medical University. RESULTS: The study revealed very frequent low back complaints among the nurses. The application of simple physical exercises proved to be effective, but not all the nurses followed the prescribed program of exercises. There was a positive correlation between the intensity and frequency of pains, duration of employment, and the range of responsibilities. CONCLUSIONS: After completing the prescribed program of exercises a significant alleviation of back pains was observed in the study group. The exercises were recognized as efficacious, uncomplicated, and providing relief. Although over a half of nurses under study did not practice exercises twice a day, as recommended, as many as 88% of nurses are going to continue the program of kinesitherapy in the future. PMID- 16218139 TI - [Effect of metals, benzene, pesticides and ethylene oxide on the haematopoietic system]. AB - The hematopoietic system, due to intensive cells proliferation, is very sensitive to toxic substances. Many chemicals, including benzene, pesticides (dithiocarbamines), ethylene oxide and metals (mercury, cadmium, chrome, cobalt, lead, aluminum) exert their toxic effect on the hematopoietic system. Exposure to each of these substances may occur in the work place due to environmental pollution and in municipal or residential areas. Exposure to lead, aluminum, cadmium, and benzene results in the incidence of anemia. In addition, exposure to benzene and its metabolites leads to myelodysplastic syndromes, leukemia, lymphomas and bone marrow aplasia. Ethylene oxide induces neoplasm of the hematopoietic system and lymphomas, especially non-Hodgkin lymphoma. Arsenic compounds act like immunosuppressants. Mercury and chrome affect the immune system by immunosuppression and by evoking autoimmune reactions. Dithiocarbamates are suspected to induce leukemia. An analysis of the pathophysiology of individual substances reveal universal toxic mechanisms. In this paper, the authors discuss the pathomechanism of toxic effects of the aforesaid chemicals on the haematopoietic system and peripheral blood cells from the viewpoint of mutagenesis, apoptosis, myelotoxicity, anemia, immunomodulation, and individual sensitivity. PMID- 16218140 TI - [Does exogenous melatonin prove to be effective in the prevention and treatment of pathologies associated with shift and night work?]. AB - The current data concerning the efficacy of exogenous melatonin treatment, especially with respect to night-shift workers, are presented. Therapy of sleep disorders experienced by night-shift workers is not always successful. To assess the results of melatonin therapy in shift workers, it is essential to perform extensive analyses of numerous physiological functions that may be affected by this work system as well as of possible late health effects in this group of workers. It is also necessary to establish indications and conditions for this kind of therapy. We may suspect that both night-shift work tolerance and efficacy of melatonin therapy are different in individual workers, and thus the choice of therapy should be based on a thorough examination of each worker's psycho-somatic conditions. PMID- 16218141 TI - Nursing television. Definitely not reality! PMID- 16218142 TI - Conscientious objection. Do nurses have the right to refuse to provide care? PMID- 16218144 TI - Near-term but still a preemie. PMID- 16218145 TI - Breast pain. Diagnosis & treatment. PMID- 16218146 TI - Building a breastfeeding center of excellence. PMID- 16218147 TI - Multicultural issues in perinatal loss. PMID- 16218148 TI - Patient safety issues in a tertiary care hospital's labor and delivery unit. PMID- 16218149 TI - Have your teenagers had their calcium today? PMID- 16218150 TI - The "wow!" factor is you. PMID- 16218151 TI - Every woman expanding nurse distributor base. PMID- 16218152 TI - Caring for the near-term infant. PMID- 16218153 TI - If I hadn't asked... PMID- 16218154 TI - The proposed Assisted Dying Bill in the UK. PMID- 16218155 TI - A day in the life of European palliative care. PMID- 16218156 TI - The House of Lords Select Committee on the Assisted Dying for the Terminally III Bill: implications for specialist palliative care. AB - The Assisted Dying for the Terminally III Bill proposed to legalise both euthanasia and physician-assisted suicide for those with a terminal illness in the UK. A House of Lords Select Committee was convened to scrutinise this Bill and has recently published its report, which will be debated in Parliament on October 10th 2005. The written and oral evidence submitted to the Select Committee represented a wide range of views on 'assisted dying'. Much of the evidence from those countries which have legalised euthanasia/physician-assisted suicide (The Netherlands, Belgium, Switzerland and Oregon, USA) dealt with the practicalities of ending life, and the legal procedures and safeguards instigated in these countries. All the written and oral evidence in the public domain was scrutinised by the authors whilst the Select Committee was sitting. We have extracted those themes relevant to specialist palliative care practice and present them in this paper. We hope that this will provide a useful resource to inform the forthcoming public debate on assisted dying. The evidence of harms inherent in making such a change in the law, as presented to the Select Committee, has moved all three authors to oppose a change in the law. PMID- 16218157 TI - Attitudes towards, and wishes for, euthanasia in advanced cancer patients at a palliative medicine unit. AB - BACKGROUND: Most studies on attitudes towards euthanasia and physician-assisted suicide (PAS) have been conducted in healthy populations. The aim of this study is to explore and describe attitudes towards, and wishes for, euthanasia/PAS in cancer patients with short life expectancy. METHOD: Semi-structured interviews with 18 cancer patients with a life expectancy of less than nine months. All patients were recruited from an inpatient palliative medicine unit. RESULTS: Patients holding a positive attitude towards euthanasia/PAS do not necessarily want euthanasia/PAS for themselves. Wishes are different from requests for euthanasia/PAS. Fear of future pain and a painful death were the main reasons given for a possible wish for euthanasia/PAS. Worries about minimal quality of life and lack of hope also contributed to such thoughts. Wishes for euthanasia/PAS were hypothetical; they were future oriented and with a prerequisite that intense pain, lack of quality of life and/or hope had to be present. Additionally, wishes were fluctuating and ambivalent. CONCLUSION: The wish to die in these patients does not seem to be constant. Rather, this wish is more appropriately seen as an ambivalent and fluctuating mental 'solution' for the future. Health care providers should be aware of this when responding to utterances regarding euthanasia/PAS. PMID- 16218158 TI - Challenging the framework for evidence in palliative care research. AB - This paper examines the debate about best evidence within the public health literature and proposes that similar arguments and concerns exist with respect to use of current evidence-based approaches to implementing research and evaluating the literature in palliative care. Whilst randomized controlled trials (RCTs) remain the gold standard and are appropriate in many instances of palliative care research, there is a need for an alternate research design framework that incorporates contextual and compositional effects pertinent to palliative care research. A framework, entitled Equity-Based Evidence, is discussed as an approach to evidence-based knowledge development in palliative care. PMID- 16218159 TI - A validation study of a pain classification system for advanced cancer patients using content experts: the Edmonton Classification System for Cancer Pain. AB - The purpose of this study was to gather construct validity evidence for a pain classification system for advanced cancer patients using content experts. Two expert panels, representing regional (Panel A, n = 18) and national/international (Panel B, n = 52) palliative medicine and pain specialists, were purposefully selected to participate in a modified Delphi survey technique, to evaluate an existing pain classification system, the Revised Edmonton Staging System (rESS). Each panel participated in two survey rounds, with response rates of 67% (Panel A, Round 1), 39% (Panel A, Round 2), 56% (Panel B, Round 1) and 64% (Panel B, Round 2). The rESS consists of five features: mechanism of pain, incidental pain, psychological distress, addictive behavior and cognitive function. Most participants either agreed or strongly agreed with including the five existing rESS features in a pain classification system, ranging from 67% (Panel A, cognitive function) to 100% (Panel B, mechanism of pain). Most participants suggested keeping the current definitions for these features, with some revisions. Based on participant feedback, definitions for incidental pain, psychological distress, addictive behavior and cognitive function were revised, including the development of guidelines for use. To reflect its intended use as a classification system, the name of the instrument was changed to the Edmonton Classification System for Cancer Pain (ECS-CP). PMID- 16218160 TI - Pain and pain treatments in European palliative care units. A cross sectional survey from the European Association for Palliative Care Research Network. AB - The Research Network of the European Association for Palliative Care (EAPC) performed a survey of 3030 cancer patients from 143 palliative care centres in 21 European countries. The survey addressed pain intensity and the use of non-opioid analgesics, adjuvant analgesics and opioids. Patients were treated with analgesics corresponding to the WHO pain ladder step I (n = 855), step II (n = 509) and step III (n = 1589). The investigators assessed 32% of the patients as having moderate or severe pain. In general there were small differences between pain intensities across different countries. Cancer primary sites and the presence of metastasis had only minor influences on pain intensity. The most frequently used non-opioid analgesics were NSAIDs (26%) and paracetamol (23%). Adjuvant analgesics or co-analgesics used by >1% of the patients were corticosteroids (39%), tricylic antidepressants (11%), gabapentin (5%), bisphosphonates (4%), clonazepam (2%), carbamazepine (4%) and phenytoin (2%). The use of non-opioid analgesics and co-analgesics varied widely between countries. Opioids administered for mild to moderate pain were codeine (8%), tramadol (8%), dextropropoxyphene (5%) and dihydrocodeine (2%). Morphine was the most frequently used opioid for moderate to severe pain (oral normal release morphine: 21%; oral sustained-release morphine: 19%; i.v. or s.c. morphine: 10%). Other opioids for moderate to severe pain were transdermal fentanyl (14%), oxycodone (4%), methadone (2%), diamorphine (2%) and hydromorphone (1%). We observed large variations in the use of opioids across countries. Finally, we observed that only a minority of the patients who used morphine needed very high doses. PMID- 16218161 TI - The healthcare needs of chronic obstructive pulmonary disease patients in the last year of life. AB - Chronic obstructive pulmonary disease (COPD) causes almost as many deaths as lung cancer, yet evidence about the impact of COPD in the latter stages of illness is limited. We assessed the healthcare needs of COPD patients in the last year of life through a retrospective survey of the informants of 399 deaths from COPD in four London health authorities between January and May 2001. We assessed symptoms, day to day functioning, contact with health and social services, formal and informal help with personal care, information received and place of death. We obtained data on 209 (52%) deceased subjects (55% male), average age at death was 76.8 years. Based on the reports of informants of the deaths: 98% were breathless all the time or sometimes in the last year of life; other symptoms reported all the time or sometimes included fatigue or weakness (96%), low mood (77%) and pain (70%); breathlessness was partly relieved in over 50% of those treated; control of other symptoms was poor, with low mood relieved in 8% and no treatment for low mood received by 82% of sufferers; 41% left the house less than once a month or never; 47% were admitted to hospital at least twice in the last year of life; 51% received regular check-ups for their chest; 36% had check-ups by a hospital consultant; 35% saw their general practitioner (GP) less than once every three months or never; 63% knew they might die; 67% died in hospital. Patients who died from COPD lacked surveillance and received inadequate services from primary and secondary care in the year before they died. The absence of palliative care services highlights the need for research into appropriate models of care to address uncontrolled symptoms, information provision and end of life planning PMID- 16218162 TI - Preferences for place of care and place of death among informal caregivers of the terminally ill. AB - OBJECTIVES: (1) To determine informal caregivers perceptions about place of care and place of death; and (2) to identify variables associated with a home death among terminally ill individuals who received in-home support services in a publicly funded home care system. PARTICIPANTS AND DESIGN: A total of 216 informal caregivers participated in a bereavement interview. Data collection included care recipient and informal caregiver characteristics, the use of and satisfaction with community services, and preferences about place of death. RESULTS: Most caregivers reported that they and the care recipient had a preferred place of death (77 and 68%, respectively) with over 63% reporting home as the preferred place of death. Caregivers had a greater preference for an institutional death (14%) than care recipients (4.7%). While 30% of care recipients did not die in their preferred location, most caregivers (92%) felt, in retrospect, that where the care recipient died was the appropriate place of death. Most caregivers reported being satisfied with the care that was provided. The odds of dying at home were greater when the care recipient stated a preference for place of death (OR: 2.92; 95% CI: 1.25, 6.85), and the family physician made home visits during the care recipients last month of life (Univariate odds ratios (OR): 4.42; 95% CI: 1.46, 13.36). DISCUSSION: The ethic of self-control and choice for the care recipient must be balanced with consideration for the well being of the informal caregiver and responsiveness of the community service system. PMID- 16218163 TI - Palliative medicine: too easy to bury our mistakes? PMID- 16218164 TI - Opioid cost: a global problem. PMID- 16218165 TI - The palliative medicine SpR training in general practice. PMID- 16218166 TI - A pilot study to assess the effectiveness of a palliative care clinic in improving the quality of life for patients with severe heart failure. PMID- 16218167 TI - Zidovudine administration during pregnancy and mitochondrial disease in the offspring. AB - Prophylactic administration of zidovudine (ZDV) to mother-child pairs reduces HIV transmission. ZDV can impair mitochondrial (mt) DNA polymerase gamma, leading to mtDNA depletion. Signs of mitochondrial dysfunction have been observed in a few children with prenatal exposure to nucleoside analogues, although no mtDNA depletion was demonstrated. Other studies failed to confirm mitochondrial disorders in children who were exposed to antiretroviral agents in utero. A child, whose HIV-infected mother received ZDV from the fourth month of pregnancy, developed neonatal encephalomyopathy, anaemia and hyperlactataemia. At 2 weeks of age, a muscle biopsy exhibited red-ragged-like fibres, proliferation of abnormal mitochondria and a 90% depletion of mtDNA without qualitative abnormalities. At 6 months, the depletion was less profound (about 50% of normal values). Severe psychomotor delay and visual disturbances persisted at 30 months, but they were greatly reduced at 5-year follow-up. These laboratory and clinical findings clearly demonstrated that mtDNA alteration was acquired and not consequent to an inherited disorder. Fetal exposure to ZDV may have caused the mtDNA depletion, which, although temporary, led to irreversible but not progressive brain damage. PMID- 16218168 TI - Cerebrospinal fluid HIV-1 infection usually responds well to antiretroviral treatment. AB - The primary objective of this retrospective study was to determine how many patients in routine practice who were treated with combination antiretroviral treatment reached HIV-1 RNA levels below 50-400 copies/ml in cerebrospinal fluid (CSF). Seventy-four antiretroviral-naive HIV-1-infected patients from five different centres in Germany, Italy, Sweden and the USA were included. Thirty nine percent of the patients had a HIV-1-associated neurological disease and 53% of the patients had AIDS. HIV-1 RNA in CSF and plasma were quantified before and after approximately 3 months of treatment. At baseline, the median value of HIV-1 RNA in CSF was 4.12 log copies/ml (interquartile range (IQR): 3.28-4.85) and it decreased to < 1.70 log copies/ml (IQR: < 1.70-2.48; P < 0.001) after in median 3 months of treatment. Seventy-six percent of the patients had HIV-1 RNA levels below the limits of detection in CSF at follow-up, and 85% reached below 400 copies/ml. In plasma, 45% of the patients had levels of HIV-1 RNA below the limits of detection at follow-up and 80% reached below 400 copies/ml. The group of patients with a neurological disease had a significantly higher CSF viral load both at baseline and at follow-up compared with the neurologically asymptomatic patients. We conclude that the central nervous system (CNS) is usually not a 'sanctuary site', difficult to reach with combination antiretroviral treatment. PMID- 16218169 TI - Long-term lamivudine monotherapy in renal-transplant recipients with hepatitis-B related cirrhosis. AB - BACKGROUND: Chronic hepatitis B virus (HBV) infection is an important cause of morbidity and mortality in renal-transplant recipients. The aim of the study was to assess the efficacy and safety of long-term lamivudine monotherapy in renal transplant recipients with HBV-related cirrhosis. METHODS: Seventeen such patients [median age: 45 years; 7 with hepatitis B e antigen (HBeAg)] received daily oral doses of 75-150 mg lamivudine for a median of 48 (range 11-81) months. All patients had baseline serum levels of HBV DNA of over 6 log10 copies per ml and of alanine transaminase (ALT) of over 1.5 times the upper normal limit (UNL). Clinical lamivudine resistance was defined as a rebound of serum HBV DNA above 5.3 log10 copies per ml and of serum ALT of over 1.5 times the UNL in patients who initially responded with HBV DNA levels of less than 5.3 log10 copies per ml and normal ALT values. Controls were 14 renal-transplant patients (median age 44 years; 3 with HBeAg) with HBV-related cirrhosis, naive to any anti-HBV therapy, followed for 58 months (4-135). RESULTS: Thirteen (77%) treated patients had a persistent response throughout the study period, including three (18%) who developed genotypic resistance, compared with none of the untreated controls (77% versus 0%, P < 0.0001). Four (23%) developed clinical resistance. Two of three patients with initially decompensated cirrhosis had a durable response and clinical improvement compared with the transient responder, whose liver function worsened following lamivudine resistance. Two responders developed chronic rejection requiring chronic haemodialysis. Overall, one treated patient developed liver-related complications, compared with eight untreated controls (6% versus 57%, P < 0.01). CONCLUSIONS: Most renal-transplant patients treated with lamivudine achieved a rapid and durable suppression of HBV, which substantially lowered the risk of liver decompensation and death. PMID- 16218170 TI - The prevalence, clinical features and response to antiviral therapy of patients with chronic hepatitis C who are seropositive for liver-kidney microsome type 1 antibodies. AB - BACKGROUND: Antibodies to liver-kidney microsome type 1 (anti-LKM-1), which are a marker of autoimmune hepatitis, are found in a minority of patients with chronic hepatitis C virus (HCV) infection. Whether interferon/ribavirin therapy is safe and effective in these patients is unclear. AIM: To describe the prevalence, clinical features and response to interferon/ribavirin therapy of anti-LKM-1 seropositive patients with chronic hepatitis C. PATIENTS AND METHODS: All anti LKM-1 seropositive patients with chronic hepatitis C who between 1997 and 2002 underwent a diagnostic liver biopsy at the Liver Center Maggiore Hospital, Milan, were studied. Serum HCV RNA was tested by in-house PCR with a limit sensitivity of 50 IU/ml. Tissue antibodies were assessed by indirect immunofluorescence on cryostat sections from rat liver, kidney and stomach. Liver biopsies were graded and staged by the Ishak score. Autoimmune hepatitis was defined according to the International Autoimmune Hepatitis Grading (IAHG) score. RESULTS: Forty-eight (1.8%) of 2675 HCV patients circulated anti-LKM-1 (30 females, 55 years of age). Twenty-eight had genotype 2, 18 genotype 1, and two genotype 3. Aminotransferase levels had been high for 23 + 12 years, on average. Using IAHG, autoimmune hepatitis was excluded in 44 patients (92%) and found to be probable in 4 patients (8%). Chronic hepatitis was histologically mild in 34 patients (70%), moderate to severe in 7 patients (15%) and with cirrhosis in 7 patients (15%). A sustained virological response (SVR) was achieved in 20 of the 27 patients who received interferon/ribavirin (13 genotype 2c with 87% SVR, and 7 genotype 1b with 58% SVR). None of the patients had serum aminotransferases, immunoglobulins or anti-LKM-1 levels flaring following therapy. CONCLUSIONS: LKM-1 antibodies rarely occur in patients with chronic hepatitis C and do not predict autoimmune hepatitis, interferon/ribavirin hyporesponsiveness or immune-related reactions to therapy. PMID- 16218171 TI - Two-year interferon therapy with or without ribavirin in chronic delta hepatitis. AB - The only beneficial agent for the treatment of chronic delta hepatitis (CDH) is interferon (IFN). However, there is no consensus on the best dosage or duration of IFN therapy. As ribavirin (RBV) increases the sustained response when added to IFN in chronic hepatitis C, probably because of its immunomodulatory effect, we aimed to investigate the efficacy of 2-year IFN treatment and whether RBV had any additive effect to IFN in CDH. METHODS: Patients (n = 31) with CDH were randomized with a 1:2 ratio as 10 patients (3 females/7 males, age 39 +/- 9) receiving IFN monotherapy (9 MU IFN-alpha2a three times weekly) and 21 patients (8 females/13 males, age 38 +/- 11) receiving IFN plus RBV for 2 years (IFN at the same dosage and RBV at 1000-1200 mg/day). Alanine transferase normalization and hepatitis delta virus (HDV) RNA negativity at the end of treatment and at the end of the follow-up period (at least 6 months following 2-year treatment) were primary endpoints of the study. In addition, virological response and biochemical response were determined separately. RESULTS: Eight of 31 patients (25%) had cirrhosis in liver biopsies. Six patients from the IFN monotherapy group and 12 patients from the combination group had biochemical response. Five patients from the IFN monotherapy group and 11 patients from the combination group had virological response at the end of therapy. Two patients from the IFN group and five patients from the combination group had sustained biochemical response at the end of the follow-up period. Hepatitis B virus (HBV) activations with HBV DNA positivity were observed in two patients (one from the IFN monotherapy group, one from the combination group). Two patients (20%) in the IFN group and five patients (23.5%) in IFN plus RBV group remained as virological responders at the end of the follow-up period (P > 0.05). None of the patients with liver cirrhosis were responsive at the end of the follow-up period. CONCLUSION: Almost 20% of the patients with CDH were responsive to 2-year IFN treatment at the end of the follow-up period and no additional effect of RBV was observed. Patients with advanced liver disease failed to respond to treatment. PMID- 16218172 TI - Selection of hepatitis B virus polymerase mutations in HIV-coinfected patients treated with tenofovir. AB - BACKGROUND: Tenofovir (TDF) is an adenosine nucleotide analogue that has been approved for the treatment of HIV-1 infection. It also shows activity against hepatitis B virus (HBV) in patients with or without lamivudine (LAM)-associated mutations. Development of clinical or virological HBV breakthrough during TDF therapy has not been reported so far. The aim of this study was to analyse the HBV polymerase (pol) from HIV/HBV-coinfected patients with detectable serum levels of HBV DNA during treatment with TDF for longer than 6 months. METHODS: The HBV pol was sequenced from 43 patient's serum before and during TDF therapy. Phenotypic analyses were performed using HBV replication-competent plasmids carrying unique mutations selected under TDF therapy. RESULTS: Mean exposure to LAM was 35.3 +/- 27.5 months and to TDF 11.2 +/- 6.7 months. Genotypic analyses from 21 of the patients revealed LAM-associated mutations, and a further two patients developed a novel mutation, rtA194T, along with LAM-resistance associated mutations. Phenotypic analyses revealed that constructs harbouring rtA194T combined with rtL180M and rtM204V displayed an over 10-fold increase in the IC50 for TDF compared with the wild type. CONCLUSION: The selection of HBV mutations in HBV/HIV-coinfected patients failing TDF therapy is an unlikely event within the first 12 months of treatment. However, HBV from two of the 43 patients treated with TDF for more than 12 months was found to contain one novel mutation located distal to the catalytic site of the HBV pol. In vitro, rtA194T conferred a reduced susceptibility to TDF in the presence of LAM-associated mutations. PMID- 16218174 TI - Prostratin and bortezomib are novel inducers of latent Kaposi's sarcoma associated herpesvirus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) establishes latent infections in lymphocytes and endothelial cells, and latent infection is closely linked to tumorigenesis. As few viral markers are expressed during latency, compounds that can safely and efficiently increase lytic gene expression in vivo have been sought. We have found that the non-tumour-promoting phorbol ester prostratin and the proteasome inhibitor bortezomib induce immediate-early, early and late KSHV gene expression from two lymphoma cell lines in vitro. Their ability to induce lytic gene expression supports a role for phorbol-ester and proteasome-regulated signalling pathways in KSHV reactivation and prompts further investigation of prostratin and bortezomib as therapeutic agents for KSHV-associated malignancies. PMID- 16218173 TI - A randomized trial to evaluate lopinavir/ritonavir versus saquinavir/ritonavir in HIV-1-infected patients: the MaxCmin2 trial. AB - OBJECTIVE: To assess the rate of protocol-defined treatment failure and safety of lopinavir/ritonavir (LPV/r) and saquinavir/ritonavir (SAQ/r). DESIGN: Open-label, prospective, randomized (1:1), international multi-centre trial. METHODS: Adult HIV-1-infected patients were assigned LPV/r 400/100 mg twice daily or SAQ/r 1000/100 mg twice daily with two or more nucleoside reverse transcriptase inhibitors (NRTIs)/non-NRTIs. All patients, whether on or off the assigned treatment, were followed for 48 weeks. RESULTS: Of 339 randomized patients, 324 initiated assigned treatment (intention-to-treat/exposed [ITT/e] population). At 48 weeks, treatment failure occurred in 29/163 (18%) and 53/161 (33%) of patients in the LPV/r and SAQ/r arms, respectively (ITT/e, P = 0.002, log rank test). In an analysis that also considered those patients who discontinued treatment as having failed treatment (ITT/e/discontinuation = failure), 40/161 (25%) LPV/r treated individuals versus 63/161 (39%) SAQ/R-treated individuals failed treatment (P = 0.005, log rank test). Discontinuation of the assigned treatment occurred in 23/163 (14%) patients in the LPV/r-treated group, compared with 48/161 (300%) in the SAQ/r-treated group (ITT/e; P = 0.001). The primary reasons for premature discontinuation were non-fatal adverse events (LPV/r: 12/163; SAQ/r: 21/161) and patients' choice (LPV/r: 7/163; SAQ/r: 8/161). In the on treatment analysis of time to treatment failure, no difference was observed between the two arms (P = 0.27, log rank test). CONCLUSION: LPV/r had better antiretroviral effects compared with SAQ/r at the doses and in the formulations studied. This may have been a result of patients' preferences and ability to adhere to assigned therapy, rather than a result of differences in the intrinsic potency of the study protease inhibitors. PMID- 16218175 TI - Comparison of three different interventions for the correction of HIV-associated facial lipoatrophy: a prospective study. AB - OBJECTIVE: To compare autologous fat transfer (AFT), injections of reabsorbable [polylactic acid (PLA)] and non-reabsorbable [polyacrylamide hydrogel (PAAG)] filler materials for the treatment of HIV-related facial lipoatrophy. DESIGN AND METHODS: Eligible individuals with enough residual subcutaneous fat in the abdomen or in the dorso-cervical region were offered AFT surgery. Other individuals were blindly assigned to two different surgical teams, who administered a set of PLA or PAAG injections every 4 weeks. The primary endpoint was the measurement of Bichat's fat pad region, determined by the result of dermal plus subcutaneous thickness. Secondary endpoints included body image evaluation (determined by ABCD questionnaire), facial aesthetic satisfaction (determined by Visual Analogue Scale), and aesthetic pre- and post-picture comparisons by independent reviewers. All variables were measured at baseline and at 24 weeks after the last treatment session. RESULTS: Twenty-four individuals received AFT and 35 were selectively randomized to PLA (20) or PAAG (15) infiltrations. PLA and PAAG groups received a mean of 5 and 6 injections respectively (P = NS). The mean change in dermal and subcutaneous thickness was 3.3 +/- 4.1 mm, 3.5 +/- 4.0 mm; 2.1 +/- 3.0 mm (P = 0.687), respectively. The mean change in ABCD score result was poorer in the AFT arm, but there were no other differences in other measured factors. Four serious adverse events were documented in the AFT arm only. CONCLUSIONS: All three interventional techniques were highly effective in improving the aesthetic satisfaction of the patients. Longer follow-up is necessary to determine the most durable and suitable treatment. PMID- 16218176 TI - A prospective study of efficacy and safety of once-daily saquinavir/ritonavir plus two nucleoside reverse transcriptase inhibitors in treatment-naive Thai patients. AB - OBJECTIVE: To assess the efficacy and safety of first-line treatment with once daily saquinavir/ritonavir with two nucleoside reverse transcriptase inhibitors (NRTIs), as induction therapy before enrollment in a randomized trial of structured treatment interruption strategies. DESIGN: Two-hundred antiretroviral naive patients with CD4+ cell counts between 200-350 at screening were enrolled in this open-label 24week study. METHODS: Patients were followed up every 8 weeks for CD4+ cells, HIV RNA, and clinical and laboratory toxicities. RESULTS: Two hundred patients were enrolled with median baseline CD4+ cell count of 267 cells/microl and HIV RNA 50 118 (4.7 log10) copies/mi. After 24 weeks of treatment, 191 of 200 (96%) patients had below 400 copies/ml HIV RNA, with 177/200 (89%) below 50 copies/ml (intent to treat, missing equals failure method), with a median rise in CD4+ cell count of 122 cells/microl. There was no significant correlation between the minimum concentration of saquinavir and HIV RNA reductions at week 8 (P = 0.957) or absolute HIV RNA at week 24 (P = 0.77). CONCLUSION: First-line highly active antiretroviral therapy (HAART) with once daily saquinavir/ritonavir plus two NRTIs showed strong antiviral efficacy over 24 weeks, and should be evaluated in larger prospective randomized clinical trials. PMID- 16218177 TI - The impact of haematopoietic growth factors on the management and efficacy of antiviral treatment in patients with hepatitis C virus. AB - AIM: To evaluate the benefits of haematopoietic growth factors (HGFs) during the treatment of chronic hepatitis C virus (HCV) infection with severe haematotoxicity. METHODS: This was a 1-year retrospective study of HCV-positive patients receiving pegylated interferon and ribavirin. Patients received different HGFs, depending on certain criteria: they received erythropoietin (EPO) when their haemoglobin (Hb) levels were less than 10 g/dl and granulocyte colony stimulating factor (G-CSF) when their neutrophil count was less than 750 cells/mm3. Haematological data, adherence and virological response were analysed and compared according to HGF use. RESULTS: In total, 132 patients were studied and 31 (23.5%) required HGF. Under multivariate analysis, baseline Hb levels of less than 13g/dl or a drop in Hb levels of over 2% per week predicted severe anaemia, and a baseline neutrophil count under 2900/mm3 predicted severe neutropaenia. HGF administration restored Hb values and the neutrophil count to above 10 g/dl and 1500 cells/mm3, respectively, in all 31 patients. Adherence to antiviral treatment was achieved in 25% of patients versus 58% of controls without severe haematotoxicity. The primary and sustained virological response did not differ statistically between HGF support and the control group (61% versus 57% and 32% versus 39%, respectively). CONCLUSION: HGF administration counteracts the severe haematological adverse effects which occur during antiviral therapy and maintains the rate of sustained response. PMID- 16218178 TI - Nevirapine concentration in cervicovaginal and oropharyngeal secretions after single-dose administration to the mother. PMID- 16218179 TI - [Animal health and animal health care concepts changing over time]. AB - The term "animal health", in particular applied to herds and flocks of food animals, needs to be newly defined in the light of the growing societal concerns with modern husbandry systems and animal welfare. This paper deals with a definition of animal health, which is extended to the well being of the animals and to pre-harvest food safety aspects. The future activities of food animal veterinarians have to meet the new demands and societal expectations discussed in this paper. PMID- 16218180 TI - [Proposal for changing the spot checking examination regarding the control of the maintenance of freedom from Aujeszky's disease (AD)]. AB - In Germany the last outbreak of Aujeszky's disease (AD) in a holding of domestic pigs had been officially confirmed in the year 2000, and, since February 2003 Germany is officially confirmed free of AD. Since this time the AD free status is monitored according to chapter II of the annex of the regulation for the protection against Aujeszky's disease. As an alternative monitoring protocol two variants of a two step random sampling scheme is proposed which reduces the sample size from a complete investigation of all domestic pig holdings to a sample of pig fattening and pig breeding farms. Within the random selected farms, pigs are sampled as usual according to chapter II of the annex of the regulation for the protection against Aujeszky's disease. In this paper, the measures to be taken in case of a positive AD result are discussed: In case of conformation of an AD suspicion it is proposed to establish a restriction area. In addition measures based on regionalisation are proposed in order to maintain the AD free status for areas outside established restriction areas. The proposed monitoring schemes potentiate an enormous reduction of sampling costs, but call for a revision of the regulation for the protection against Aujeszky's disease. PMID- 16218182 TI - [Mange eradication in piglet producing farms in the Weser-Ems region]. AB - A variety of reports on mange eradication exist in international literature. Adequate programs are yet to be established in Germany. The commonly used examination of skin scrapings proved to be insufficient. Due to reliable results the pig health service is using Sarcoptes-ELISA (AFOSA GmbH). In the years 2001 and 2002 a Sacoptes suis eradication was accomplished in two sow farms in the Weser-Ems region and since continuously monitored clinical examinations have been used, as well as ELISA testing and skin scrapings. After two years of monitoring a certificate on "unsuspicious for mange" was given to both farms in November 2003 and June 2004. The practical approach will be described. This Sarcoptes eradication is possible even in areas with intensive pig production. When setting up or expanding high health breeding or multiplying herds "unsuspicious for mange" should be one of the important health targets. The practical results with the use of the Sarcoptes-ELISA for monitoring sow farms are promising. PMID- 16218181 TI - [Methods for population-based herd sanitation: an overview]. AB - There is a considerable number of methods available for the development, maintenance and surveillance of specific-pathogen-free herds. The selection of a method for a specific remediation project is primarily dependent on the goal, the epidemiology of the agent to be eradicated, the feasibility and the available resources. At the beginning of the remediation programme, methods are required for the classification of the herd status. Clinical and analytical diagnostic tests may be used for individual herds or in an entire area. The interpretation of test results applied within a population deserves special attention. A series of techniques are available - alone or in combination - to remediate a herd that was classified as being infected: systematic treatment, vaccination, elimination of infected individual animals ("test and remove"), partial depopulation or total population (stamping out). These strategies are typically accompanied by cleaning and disinfection. After the remediation, the status of the herd needs to be certified and maintained. For the documentation of the status, monitoring and surveillance strategies such as clinical follow-up visits, notification systems and random sampling can be applied. When planning random surveys, statistical and epidemiological considerations are relevant. For the sustainable maintenance of the pathogen-free status, biosecurity measures at the individual farm level, but also broader approaches such as import restrictions, early warning systems and surveillance programmes are needed. General awareness regarding unusual clinical signs needs to be promoted among farmers and veterinarians. The longer a disease or an agent is absent from a herd or region, the lower becomes the risk awareness. In such a situation, mistakes can occur leading to an increased risk of re-infection. PMID- 16218183 TI - [Do we need pathogen-free livestock for food safety?]. AB - The reduction of the entry of pathogens in the food chain is an important premise in improving the safety of food of animal origin. Since food animals are the main reservoir for pathogens, the prevalence of pathogens in livestock is of great importance. Unfortunately, classical measures to improve animal health can not exclude the presence of the main food-borne zoonotic agents (Salmonella, Campylobacter, Listeria monocytogenes, verotoxinogene Escherichia coli, Toxoplasma gondii) in "clinically healthy" animals. Absence of pathogens in livestock must be regarded as an unrealistic aim. To achieve an effective improval of product safety through logistical, hygienic or technological measures both farmers and food producers need valid data about the prevalence of pathogens in livestock, in each animal and in raw material of animal origin, respectively. Yet available data are insufficient. New European directives and regulations concerning monitoring and control of zoonosis demand the systematic control of presence of zoonotic agents in livestock and later steps of the food chain. PMID- 16218184 TI - [Observations on the control and eradication of paratuberculosis in dairy herds]. AB - Johne's disease is widely seen in dairy herds in Germany. Estimates based primarily on epidemiological surveys in neighbouring states assume that 5 to 15 % of German herds are infected. In the past three years several authors have reported that the causative agent of Johne's disease, Mycobacterium avium subspecies paratuberculosis (MAP), is found ubiquitously in the environment and can be isolated from a number of different animals, including non ruminants. These results imply that MAP should be considered an environmental pathogen. Based on this assumption a concept for control and eradication of Johne's disease is presented aiming at minimizing the future spread of disease and reducing environmental contamination with the pathogen at low costs. The concept includes the classification of herds based on an bulk milk ELISA followed by a robot compatible bulk milk PCR in ELISA-positive herds only. Due to the comparatively low costs combined with the high specificity of the approach a detection of heavily infected herds ("tip of the iceberg") all over the country would be possible; based on the eradication of strong shedders in these herds the input of MAP into the environment would be reduced considerably. PMID- 16218185 TI - [Experiences with the eradication of the bovine herpes virus 1 in Lower Saxony]. AB - In this paper first the general cascade of animal disease eradication from the voluntary control programme up to culling of infected animals is described. Afterwards the development of the eradication of Bovine Herpes Virus in Lower Saxony is illustrated. Obvious the number of farms which are not eradicating decreases since the testing on BHV1 is obligate in Germany. In 2004 60% of the cattle holdings were free from BHV1. At least the measures which will be taken in Lower Saxony to increase the success of eradication are explained. PMID- 16218186 TI - [Specified pathogen-free poultry flocks: the current situation]. AB - Before the implementation of strategies to establish specified pathogen-free commercial poultry flocks, the ultimate goals need to be identified: 1) consumer protection by minimizing the risk for zoonotic diseases and food-borne pathogens, and/or 2) animal health protection against primary and secondary pathogens. The success for the establishment of specific pathogen-free poultry flocks depends on the biological characteristics, the epidemiological distribution and the transmission route of each pathogen. For zoonotic pathogens such as Salmonella Typhimurium, Salmonella Enteritidis, Campylobacter jejuni or the high pathogenic avian influenza virus, eradication has to be ultimate goal. Despite tremendous control efforts in field, only partial control of these pathogens has been achieved so far. In the future it will be necessary to continue these eradication efforts by combining optimized hygiene programs at all production levels with intensive monitoring and immunoprophylaxis. For primary pathogens affecting the health condition of poultry without known zoonotic potential, such as Salmonella Gallinarum, avian Mycoplasma or leucosis virus, specified pathogen free flocks have been established on the parent and grandparent level. In order to achieve a status free of these pathogens, rigid hygiene control, especially on the hatchery level and monitoring programs combined with elimination of pathogen- and antibody positive birds were implemented. Nevertheless, the economically most important diseases of modern poultry production are of multifactorial origin. Ubiquitous secondary pathogens in combination with insufficient management or immunosuppressive agents induce great economic losses for the poultry producers. These secondary pathogens can not be eliminated due to their ubiquitous distribution. In the future only a reduction of these factorial diseases will be possible combining hygiene management and optimization of poultry husbandry. For the establishment of specified pathogen free poultry flocks in the field, risk analysis is necessary and the structure of poultry production has to be considered before and eradication program can be carried out successfully. PMID- 16218187 TI - [Safeguarding herds from the animal hygiene point of view]. AB - One of the most important concepts for the protection of herd health is the implementation of structural and organisational measures to prevent infective agents and other adverse compounds from entering the farm. Safeguarding health, well-being and production efficiency is part of the overall management concept of hygiene in animal production systems. This paper presents an overview of the most important rules and recommendations to protect livestock production facilities from the intrusion of infectious pathogens, beginning with the right choice of the site for the farm and the animal housing ("safe distances" to neighbouring farm animal houses), solid fencing and control and disinfecting places at the entrance gate. The traffic of vehicles and people transporting animals, feedstuff, equipment and slurry or manure to and from the facility should be reduced to a minimum. Fallen animals should be stored in separate and safe containers until removed by specialised companies. Regular control of rodents, insects and wild birds is crucial to avoid the transfer of infectious agents from farm to farm and between herds within a farm. Equally important factors are the health status of personnel to avoid transmission of zoonotic diseases, the application of the all-in-all-out system and a strict cleaning and disinfecting regime. The internal and external organisational measures for preventing the spread of infections in animal production will gain increasing importance in the future because the farm animal producer bears the responsibility for the production of safe and healthy food at the primary segment of the food chain. Increasing restrictions on the use of veterinary drugs for food delivering animals will increase the importance of prophylaxis, prevention and protection of production units as the keys for safeguarding health, well-being and efficiency of farm animals. Only the application of strict hygiene principles in animal production will make it possible to meet the consumer demand for safe and high quality food of animal origin. PMID- 16218188 TI - The long-term culture of pig liver sinusoidal endothelial cells: the Holy Grail found. PMID- 16218189 TI - Long-term preservation of high endocytic activity in primary cultures of pig liver sinusoidal endothelial cells. AB - Together with Kupffer cells, liver sinusoidal endothelial cells (LSECs) constitute the most powerful scavenger system in the body. However, studies on LSEC function are hampered by the fact that the cells lose their scavenger ability and start deteriorating after a few days in culture. The purpose of the present study was to improve the conditions of cultivation to prolong the survival of pig LSECs in vitro. We used the high capacity receptor-mediated endocytosis of soluble waste molecules as a marker for functionally intact cells in the cultures. Compared with two commercially-, and two other media specifically designed for use with either SECs or hepatocytes from rat, our newly developed serum-free medium, DM 110/SS, devoid of any components of animal origin, was superior in maintaining the endocytic activity. Of six growth factors studied for their effect on endocytosis, basic fibroblast, and recombinant epidermal, but not vascular endothelial growth factor, were found to be most beneficial. After 8 days in DM 110/SS, LSECs maintained endocytosis via the scavenger receptor, mannose receptor, collagen alpha-chain receptor and the Fc gamma receptor. All endocytosed ligands, except for aggregated IgG were degraded in 8-day-old cultures. Using the new medium, the cells endocytosed ligands for up to 20 days, and survived for at least an additional 10 days, albeit without the high endocytic activity typical of intact LSECs. Importantly, DNA synthesis in prolonged cultures of LSECs was observed only when maintained in DM 110/SS medium. In conclusion, we describe a protocol for the maintenance of LSECs in culture for the longest period yet reported. PMID- 16218190 TI - Lamins A and C are differentially dysfunctional in autosomal dominant Emery Dreifuss muscular dystrophy. AB - Mutations in the LMNA gene, which encodes nuclear lamins A and C by alternative splicing, can give rise to Emery-Dreifuss muscular dystrophy. The mechanism by which lamins A and C separately contribute to this molecular phenotype is unknown. To address this question we examined ten LMNA mutations exogenously expressed as lamins A and C in COS-7 cells. Eight of the mutations when expressed in lamin A, exhibited a range of nuclear mislocalisation patterns. However, two mutations (T150P and delQ355) almost completely relocated exogenous lamin A from the nuclear envelope to the cytoplasm, disrupted nuclear envelope reassembly following cell division and altered the protein composition of the mid-body. In contrast, exogenously expressed DsRed2-tagged mutant lamin C constructs were only inserted into the nuclear lamina if co-expressed with any EGFP-tagged lamin A construct, except with one carrying the T150P mutation. The T150P, R527P and L530P mutations reduced the ability of lamin A, but not lamin C from binding to emerin. These data identify specific functional roles for the emerin-lamin C- and emerin-lamin A- containing protein complexes and is the first report to suggest that the A-type lamin mutations may be differentially dysfunctional for the same LMNA mutation. PMID- 16218191 TI - Quantitative imaging of subcellular calcium stores in mammalian LLC-PK1 epithelial cells undergoing mitosis by SIMS ion microscopy. AB - Quantitative 3-D total calcium gradients, representing subcellular stored calcium, were imaged with a CAMECA IMS-3f SIMS ion microscope in cryogenically prepared frozen freeze-dried LLC-PK1 cells captured in interphase and various stages of mitosis. 39K and 23Na concentrations were also measured in the same cells. Correlative optical (or SEM) and SIMS analysis of cells revealed a redistribution of the interphase Golgi calcium store in prophase and prometaphase cells. In metaphase cells, simultaneous SIMS imaging of total calcium in both the spindle and the non-spindle cytoplasm of individual cells revealed a gradual and dynamic alignment of calcium stores in both half-spindles prior to the onset of anaphase. The anaphase cells revealed the highest local total calcium concentrations in the spindle regions behind the daughter chromosomes and the lowest in the central spindle region. The pericentriolar material in telophase cells contained calcium stores. Quantitatively, a typical metaphase cell with well-aligned calcium stores in the spindle region contained 1.1 mM total calcium in each half-spindle, 0.8 mM total calcium in the non-spindle cytoplasm, and 0.5mM total calcium in the chromosomes. At the submicron scale, the distribution of total calcium was heterogeneous in the chromosomes, metaphase spindle, and non spindle cytoplasm. An increased binding of calcium to chromosomes is not a physiological requirement for chromosomal condensation in mitosis, since interphase nuclei and mitotic chromosomes contained comparable total calcium concentrations measured per unit volume. A significant reduction of total calcium in the non-spindle cytoplasm was observed in the metaphase, anaphase, and telophase cells, which is indicative of the limited storage of the releasable calcium pool in these specific stages of mitosis. Direct total calcium measurements in subcellular regions confirmed that both the spindle and the non spindle cytoplasm of metaphase cells contained inositol 1,4,5-trisphosphate (IP3) sensitive calcium stores sensitive to arginine vasopressin, thapsigargin, and calcium ionophore A23187. The dynamic alignment of calcium stores in both half spindles may be an integral part of the time-dependent process of a cell's overall preparation for exiting the metaphase stage in mammalian LLC-PK1 cells. PMID- 16218192 TI - Comparison of the effects of barbiturate, benzodiazepine and ketamine on visual evoked potentials in rabbits. AB - The aim of this study was to compare the effects of barbiturate, benzodiazepine and ketamine on flash-evoked potentials (F-VEP) in adult rabbits. A total of 36 animals were studied, 16 after pentobarbital endovenous (EV) infusion, 10 after midazolam EV administration, and 10 after ketamine EV infusion. Pentobarbital induced triphasic F-VEP, first negative (N1), second positive (P1), third negative (N2) waves, all with large amplitudes and P1 with well-defined morphology. Mean P1 latency was 33ms. Midazolam induced similar but less defined triphasic waves, with mean latency of 27ms. Ketamine induced poliphasic and poorly defined F-VEP, with mean first positive (P1) latency of 27ms. Statistical analysis showed more elongated latency for the pentobarbital group than the midazolam and ketamine groups. The results of this study suggest that the pharmacological effects of pentobarbital and midazolam on GABA neurotransmission in rabbit visual cortex may be different; another neurotransmission system, possibly cholinergic, may be involved. The ketamine effect seen in rabbit visual cortex seems to be different from pentobarbital and midazolam. PMID- 16218193 TI - Brainstem auditory evoked responses that disappear during sleep: a possible manifestation of a neurogenic vestibular evoked response. AB - OBJECTIVE: To present a case of unilateral hearing loss in which a brainstem auditory evoked potential (BAEP) disappeared during sleep on the symptomatic side, and to argue that this may actually be a manifestation of a neurogenic vestibular evoked potential (NVESTEP). MATERIAL AND METHOD: Brainstem auditory evoked potentials were performed in the standard manner. RESULT: A poorly organized response was obtained during wakefulness on the symptomatic side that resembled a BAEP. The BAEP for the right ear was better organized. During sleep, the response for the left ear disappeared. CONCLUSIONS: The poor organization of the response of the left ear compared to the right, and its disappearance during sleep, suggests that the response for the left ear was actually an NVESTEP and not a BAEP. The possibility of recording vestibular responses with auditory stimuli may have important implications for BAEP examinations performed during wakefulness that may lead to false negative results, in neonatal screening for hearing loss that use tone stimuli, and also in brain mapping using magnetic resonance imaging (MRI) and positron emission tomography (PET) relating to the auditory cortex. PMID- 16218194 TI - The double peak-to-peak analysis for determining EMG onset of muscle contraction. AB - PURPOSE: Identification of the onset of muscle contraction with EMG signal amplitude double of the baseline value (DP-P) has been recently reported for determining the temporal parameters of muscular activity. Due to its convenience, it is suitable for clinical application. However, there is a lack of report on the reliability and comparability of this method to other established methods. Therefore, this study examined the test-retest reliability of the DP-P method and compared it with an established method that used the mean + 3 standard deviations (mean + 3 s.d.) over the baseline value for muscles of the knee. METHODS: The onset of contraction of vastus medialis obliquus (VMO) and vastus lateralis (VL) of eleven able-bodied volunteers performing isometric knee extension at 50%, 75% and 100% of MVC in 30-minute and 7-day intervals were analyzed with both the DP-P and mean + 3 s.d. methods. RESULTS: The ICC for within-day measurements of DP-P method ranged from 0.64 to 0.86 and that for between-day measurements ranged from 0.63-0.81. The ICC values were higher with submaximal than maximal contractions. There was a consistent delay of about 3 ms in EMG onset detection with the DP-P as compared to the mean + 3 s.d method. CONCLUSION: The DP-P method is a reliable method for muscle onset determination but the absolute onset time of muscle contraction obtained from this method should not be directly compared with other methods such as the mean + 3 s.d. PMID- 16218195 TI - Vitamin B12 deficiency: a clinical and electrophysiological profile. AB - OBJECTIVE: The present study was conducted to evaluate the clinical and electrophysiological profile of vitamin B12 deficiency syndrome and whether a correlation exists between the disease process and the various electrophysiological parameters. METHODS: 40 patients with vitamin B12 deficiency neurological syndromes with low serum vitamin B12 and high homocysteine levels were subjected to a detailed motor and sensory nerve conduction studies and pattern reversal VER (P100), SSEP (P37) after stimulation of the posterior tibial nerve and median nerve (N 20) were obtained bilaterally. MR cervical spine was done in all the patients and MR brain in those who presented with neuro psychiatric symptoms. The patients were followed up at three months, six months and one year after treatment. RESULTS: On the basis of clinico electrophysiological profile, 31 patients had myeloneuropathy, 5 isolated myelopathy, 4 isolated neuropathy. Four of the patients with myeloneuropathy, had concomitant dementia. MR imaging revealed abnormality in 12.5% of cases. Prolongation of P37 latency was observed in 39 (97.5%) patients, N 20 latency in 22 (55%), and P100 latency in 19 (47.5%) patients. Peripheral neuropathy was seen in 18 patients; optic neuropathy in 8; and combination of peripheral and optic neuropathy in 9 patients. The peripheral neuropathy was axonal in 19, and demyelinating in 6 patients. There was a significant correlation of the duration of the disease with N 20 latency (P < 0.04). Serum vitamin B12 level correlated well with the latencies of P37 (P < 0.005) and sural SNAP (P < 0.006). On treatment, normalization of P100, MRI signal, N 20 and partial recovery of P37 latencies was seen at 6 months, 9 months and one year respectively. CONCLUSION: Differential recovery of central and peripheral syndromes was seen. This correlated with the underlying demyelinating and axonal processes, which was well reflected by the electrophysiological studies, and has an important bearing on the outcome. PMID- 16218196 TI - Tibial muscular dystrophy with late adult onset in a Spanish family. AB - PURPOSE: We report autosomal dominant distal muscular dystrophy in 5 members of a Spanish family. INTRODUCTION: This unusual muscular disorder has late adult onset and predominantly it affects the anterior compartment of the legs. This myopathy presented clinical and electromyographical characteristics, but unspecific histological findings. Early there have appeared genetical studies, the most frequently used is chromosome linkage, but it is not an absolute criterion for diagnosis, and it is not available in most hospitals. PATIENTS DESCRIPTIONS: In our cases walking difficulties appeared between the fourth and fifth decades, characterized by progressive and varied weakness with amyotrophy in the tibial anterior compartment. The electromyography confirmed the presence of a severe non inflammatory myopathy, chronic and symmetric in the pretibial muscles and of less intensity in the calf muscles. The levels of creatine phosphokinase were normal and muscle biopsy identified a chronic, unspecific lesion with important fibrosis. CONCLUSIONS: The findings, although with some phenotypical differences, were those commonly found in Markesbery-Griggs disease, tibial muscular dystrophy or late onset type 2 distal myopathy. We report a family affected by this muscular disorder, we describe the differential diagnosis and we discuss the review of the available literature. PMID- 16218197 TI - Implementation of the TMS in the early stages of Parkinson's disease. AB - 47 PD patients were investigated with the single-pulse TMS to find out changes in motor evoked potential and motor conduction related to the stage of minimal motor symptoms and its further deterioration in groups with the different clinical types of the disease. The investigation revealed a markedly longer MEP duration along with the increased number of phases, than in controls, which were bilateral and advanced despite the minimal unilateral motor symptoms. There was also increased MEP amplitude in facilitation, with a higher degree of asymmetry, compared to controls. Patients with predominant rigid clinical forms had the further MEP duration and amplitude increase proportionally to bradikinesia and rigidity in the early stages of the disease. Patients with tremor predominant forms had no further changes in the MEP duration and amplitude, but had their motor CCT decreased in the early stages. Patients with the akinetic form were characterized by the asymmetric increase in the MEP Amplitude in relaxation and motor CCT shortening. Thus, TMS allows us to diagnose early the possible central motor changes secondary to Parkinson's disease, reveals the difference in compensational capacity according to the clinical type of the disease and helps in monitoring of the severity of motor changes in early stages. PMID- 16218198 TI - Role of Blink Reflex in diagnosis of subclinical cranial neuropathy in Diabetic Mellitus type II. AB - BACKGROUND AND OBJECTIVES: Peripheral Neuropathy (PN) is one of the late complications of Diabetes Mellitus. Cranial nerves III, VII and V are among the most commonly affected in diabetic patients. Traditional Electrodiagnosis (Edx) studies are useful method for diagnosis of PN and symptomatic cranial neuropathy, and may not be useful for detecting subclinical involvement of cranial nerves. The main objective of this study is to evaluate the role of Blink Reflex (BR) for early diagnosis of cranial neuropathy in diabetic patients with PN. DESIGN: A prospective study was performed on NIDDM patients with a PN. 188 subjects were included in our study in which 142 acted as healthy subjects and 46 as diabetic patients. Patients were excluded with prior history of cranial nerve lesions, stroke, and other disease with polyneuropathy or drug-induced neuropathy. Routine nerve conduction studies were performed and only patients with PN were included in this study. RESULTS: Abnormalities were found in 54.4% of patients. R1, IR2 and CR2 were prolonged relative to healthy group. Statistically there was no significant difference in R/D ratio of patients (P = 0.201). Also there was a positive correlation between R1, IR2 and CR2 latencies with duration of diabetes and severity of polyneuropathy, but not for R/D. The greatest correlation was shown in R1 latency (69.9% abnormality). CONCLUSION: BR is a non-invasive and very useful method for evaluation and diagnosis of subclinical cranial nerve involvement in diabetic patients. PMID- 16218199 TI - Differential diagnosis of intractable lateral knee pain with progressive symptoms: a case problem. AB - This case study of an active 22 year old female who sustained a traumatic incident to the right knee focuses on the differential diagnostic process. Although early subjective complaints, clinical presentation, and diagnostic testing results were commensurate with internal derangement of the knee, appropriate medical and surgical management failed to halt the progression of pain and subsequent onset of neurological symptoms. Electrodiagnostic testing conducted 10 months post initial injury produced a definitive diagnosis of an irritative lesion of the right peroneal nerve in the region of the fibular neck. A fibular tunnel release and peroneal nerve decompression was subsequently performed after which complete symptom resolution was achieved, allowing return to full functional activities. PMID- 16218200 TI - Blink reflex in motor neuron disease. AB - The aim of the study was to analyse the diagnostic value of blink reflex (BR) in motor neuron disease (MND). We studied 25 patients with MND including amyotrophic lateral sclerosis (ALS) in 18 patients, primary muscular atrophy (PMA) in 4 patients, and primary lateral sclerosis (PLS) in 3 patients and 12 healthy volunteers. The blink reflex was obtained in typical way. In ALS group, statistically significant increase in latency and decreased amplitude of R2 responses were found compared to the control group (p < 0.05), whereas BR in patients with PLS and PMA was within normal limits. The presence of low amplitude of R2 responses in patients with ALS may suggest loss of lower brainstem neurons connecting trigeminal and facial system, and probably also decreased facilitator effect of central nervous system on reticular formation in the brainstem. PMID- 16218201 TI - Managing compliance in production-insights and comparisons in chemical pharmaceutical companies. PMID- 16218202 TI - Endotoxin in parenteral medicines may be a risk factor in severe sepsis. PMID- 16218203 TI - Pharmaceutical development of a parenteral lyophilised formulation of the investigational anticancer agent ES-285.HCl. AB - The aim of this study was to design stable parenteral pharmaceutical final products containing 25 mg and 50 mg ES-285.HCl per dosage unit for use in phase I clinical trials. ES-285.HCl drug substance was fully characterised and showed very slight solubility in water. The development of the pharmaceutical product, containing 2-hydroxypropyl-beta-cyclodextrin, is discussed in view of formulation optimisation and manufacture. The developed freeze-dried products were found stable for at least 6 months at an accelerated storage condition of 25 +/- 2 degrees C/60 +/- 5% relative humidity and for at least 12 months at the designated long term storage condition of 5 +/- 3 degrees C, in the dark. Phase I trials using ES-285.HCl 25 mg/vial and 50 mg/vial final products are currently ongoing. PMID- 16218204 TI - Evaluation of peracetic acid permeation during flash sterilization through pharmaceutical plastic polymers used in cytotoxic reconstitution units. AB - Peracetic acid (PAA) permeation in flash sterilization was studied using three different plastic infusion bags made of polypropylene and polyethylene, filled with glucose 5% or NaCl 0.9%. The pH was measured and acetic acid (AA) and PAA concentrations were made by reverse phase high-performance liquid chromatography (RP-HPLC). PAA was derivatized by oxidation of methyl tolyl sulfide (MTS) into methyl tolyl sulfoxide (MTSO) detected by ultraviolet (UV) absorbance at 230 nm. The technique has a sensitivity of 0.3 microg x L(-1) and was highly specific. Results showed that pH measurements remain constant and demonstrated the absence of PAA permeation, which was confirmed by the absence of AA permeation regardless of the brand tested, with both unwrapped and overwrapped infusion bags, when flash sterilization is applied. These results allow flash sterilization to be performed with unwrapped infusion bags without any risk of drug degradation by PAA. This makes compounding safer and easier, which improves productivity. PMID- 16218205 TI - Linking extractables and leachables in container/closure applications. AB - Establishing a link between extractables and leachables may be necessary to understand, interpret, assess, quantify, or control the interaction between a drug product and its container/closure system. This paper considers the various factors that affect the rigor with which such a linkage is established and justified. Such an assessment considers the origin and/or genesis of the leachable or extractable, enumerates situations in which extractables/leachables linkages are useful, ties such situations to the drug product's lifecycle, defines a hierarchy of linkages based on the rigor with which the linkages are established and justified, and establishes guidelines for how to determine what kind of linkage is appropriate for certain circumstances and situations. Additionally, this paper gives several examples of linkages relevant to flexible plastic drug product containers. PMID- 16218206 TI - The axis of orientation of the synlophe in the Heligmosomoidea (Nematoda, Trichostrongylina): a new approach. AB - The definition of the axis of orientation of the synlophe is modified for the Heligmosomoidea so that one or two axes may be recognized. When two axes are present, their inclinations to the sagittal axis are different on the right and left sides, and we propose to name them right axis and left axis, respectively. During the course of evolution, starting from a single oblique axis (plesiomorphic state), an independent rotation of this axis on the right and left sides may bring about a double-axis state with a different inclination on both sides (derived state). When the rotation reaches 90 degrees for both sides, the axis becomes simple once again and is superimposed to the frontal axis (most derived state). PMID- 16218207 TI - Description of Paranoplocephala yoccozi n. sp. (Cestoda: Anoplocephalidae) from the snow vole Chionomys nivalis in France, with a review of anoplocephaud cestodes of snow voles in Europe. AB - We describe Paranoplocephala yoccozi n. sp. (Cestoda: Anoplocephalidae) from the snow vole Chionomys nivalis in Bourg-Saint-Maurice, French Alps, compare it with several related species from rodents, and review the anoplocephalid cestodes of snow voles in Europe. Paranoplocephala yoccozi n. sp. is primarily distinguished from the related species by its large scolex of characteristic shape, robust neck region, and the structure of the cirrus sac, vitellarium and vagina. We show that the anoplocephalid cestodes of snow voles in Europe, representing the genera Anoplocephaloides and Paranoplocephala, include at least seven species. This fauna consists primarily of species that snow voles share with other voles inhabiting the high-mountain areas. Some of the species, including P. yoccozi n. sp., appear to have a very localized distribution, which is assumed to be a consequence of the historical fragmentation of snow vole populations. PMID- 16218208 TI - [Monogenean parasites from Clariidae (Teleostei, Siluriformes) in Cameroon: I. Description of two new species of Gyrodactylus from the Nyong basin]. AB - The study of gill and skin parasites of three fishes species belonging to the genus Clarias: C. camerunensis, C. jaensis and C. pachynema, from the Nyong basin (Cameroon), revealed the presence of two new Monogenean species of the genus Gyrodactylus Nordmann: G. camerunensis n. sp. and G. nyongensis n. sp. parasite of Clarias camerunensis, C. jaensis and C. pachynema. Gyrodactylus camerunensis n. sp. is clearly distinguished from G. nyongensis n. sp. by the smaller size of the haptor sclerites. These two new species differ also from their closest African relatives of the genus Gyrodactylus, G. rysavyi, G. clarii and G. alberti, by the morphology and size of the haptorail sclerites. Their host specificity was discussed and appeared stenoxenous (mesostenoxenous). PMID- 16218209 TI - An ultrastructural and histopathological study of Henneguya pellucida n. sp. (Myxosporea: Myxobolidae) infecting Piaractus mesopotamicus (Characidae) cultivated in Brazil. AB - During a study of myxosporean parasites of cultivated freshwater fish, a new myxosporean species, Henneguya pellucida n. sp., was discovered. Of the 120 Piaractus mesopotamicus sampled, only 10 specimens (8.3%) were infected. Yellow, round plasmodia measuring 0.5-3 mm were found in the serous membrane of the visceral cavity and in the tunica externa of the swim bladder. Sporogenesis was asynchronous, with the earliest developmental stages aligned prevailingly along the endoplasmic periphery and mature spores in the central zone. The mature spores were pear shaped (total length: 33.3 +/- 1.5 microm, mean +/- SD; width: 4.1 +/- 0.4 microm; body length: 11.4 +/- 0.3 microm; caudal process length: 24.1 +/- 1.5 microm). The polar capsules were elongated (length: 4.0 +/- 0.4 microm; width: 1.6 +/- 0.2 microm). The development of the parasite in the swim bladder produced thickening of the tunica externa and a granulomatous reaction. There was no correlation between the prevalence of the parasite and the chemical and physical characteristics of the water. Infection was recorded only in juvenile specimens ranging in size from 9.5 to 20 cm. PMID- 16218210 TI - A two-year survey on mosquitoes of Lebanon. AB - A total of 6,500 mosquitoes were identified during a two-year survey (1999-2001) in Lebanon, and these belonged to twelve species: Culex pipiens, Cx. laticinctus, Cx. mimeticus, Cx. hortensis, Cx. judaicus, Aedes aegypti, Ae. cretinus, Ochlerotatus caspius, Oc. geniculatus, Oc. pulchritarsis, Culiseta longiareolata and Anopheles claviger. Culex pipiens was the most predominant species in Lebanon, collected indoors and outdoors. It was continuously abundant and active throughout the year. Culex judaicus was a small and rare mosquito and it is reported to occur for the first time in Lebanon. On the coastal areas, Ochlerotatus caspius was very common, and proved to be a complex of species as two forms were detected. One of the vectors of malaria, Anopheles claviger, was collected from May to September, from eight sites in Lebanon. Its breeding sites were restricted to fresh, cool, and clean water in pools and wells. Most of these breeding sites were in the populated Metn County where a few indigenous cases of malaria were reported from 1997-2000. This shows that the reported malaria cases were not imported, but caused by the bites of locally infected Anopheles females. PMID- 16218211 TI - [Macrostylophora kinabaluae n. sp. (Insecta: Siphonaptera: Ceratophyllidae), a new flea from Sabah (Borneo, Eastern Malaysia)]. AB - This species, known only by a single male, is described from Mount Kinabalu and thus is recorded from the same area as Macrostylophora borneensis (Jordan, 1926), teste Traub (1972). It is distinguished from its congeneric and characterized by the absence of eriged setae on the thorax and first abdominal tergits, as well as by the shape of terminal segments and the phallosom. Macrostylophora kinabaluae was found to parasitize the Sciurid rodent Callosciurus prevostii that is widespread throughout most areas in the Malaysian subregion. PMID- 16218212 TI - Anatomy of Tunga trimamillata Pampiglione et al., 2002 (Insecta, Siphonaptera, Tungidae) and developmental phases of the gravid female. AB - This paper deals with some internal anatomical features observed in histological sections and freshly dissected mounts of Tunga trimamillata, a Siphonaptera recently discovered in Andean regions of Ecuador from several mammals, including man. It was possible to study in males and also non-gravid and gravid females, the location and anatomy of several organs not previously described for this species: the testes, epididymis, ganglia, Malpighian tubules, eyes, rectal ampulla with one of its pads and structures which could be interpreted as midgut diverticula, whose presence has not been recorded in the Siphonaptera. The process of neosomy in the female during pregnancy is illustrated by photographs of the consecutive developmental phases, taken at the stereomicroscope. Furthermore, some details of the exoskeleton, spermatheca during different phases of pregnancy of the gravid female and the presence of a foreign body (parasite?) within the haemocoel have been displayed in specimens cleared with Hoyer's medium. PMID- 16218213 TI - Antileishmanial activity of polycyclic derivatives. AB - 33 polycyclic derivatives have been studied and tested on Leishmania donovani and L. major promastigotes. Their antileishmanial activity was assessed in vitro and an assay of their cytotoxicity was realized on human myelomonocytic cell line. The reference molecules used in the assays were amphotericin B and pentamidine. Among the compounds tested, 29 possess an antileishmanial activity; 25 of those were more active against L. donovani than amphotericin B, and nine were as effective as amphotericin B against L. major. Many synthesized derivatives were more active against L. donovani than against L. major. The cytotoxicity studies have shown that among the thirty-three derivatives tested, 12 molecules have an IC50 towards THP-1 cells about equal than that reference drugs, the 21 other derivatives are much less toxic. A 3D QSAR study was undertaken and has permitted to predict activity against L. donovani and L. major and to highlight critical area to optimize activity against the two species. PMID- 16218214 TI - [Risk evaluation in sleeping sickness: a mathematical contribution]. AB - The concept of "risk" is important in epidemiology but is often used in a confused way for sleeping sickness. Using a rigorous approach resulting from mathematical modelling, new virtual entomological risk indicators and parasitological transmission indexes derived from the basic reproduction rate R0 are proposed and discussed. PMID- 16218215 TI - Postoperative changes in serum cytokines profile and nitric oxide levels in patients with cystic echinococcosis. AB - The aim of the present study was to examine serum cytokines and nitric oxide (NO) levels in patients with cystic echinococcosis (CE). 28 patients with CE were studied and all underwent surgery. Serum levels of tumour necrosis factor-alpha (TNF-alpha), interleukin IL-1beta, receptor of soluble IL-2R (sIL-2R), IL-6, IL 8, nitrate/nitrite, and C-reactive protein (CRP) were determined before and after induction of treatment. Data were compared with those obtained from 28 healthy volunteers. IL-6 was elevated in all CE patients (100%). IL-8 was increased in 11/28 (39.3%). Increased levels of IL-2R and TNF-alpha were found in a limited number of them particularly those showing cysts in the central area of the liver (5/28, 6/28). IL-1beta level was not elevated in any patient except in secondary severe CE. CRP and nitrate/nitrite levels were also increased. A positive correlation between CRP and IL-6 (r = 0.74; p < 0.001) was found confirming the link between inflammation due to CE and activation of monocytes. All patients completely recovered and the levels of the studied parameters reverted to normal levels except one patient in whom severe recurrent disease occurred two years after the first operation. These results suggest that there are different immunoregulatory events and cytokines response during CE and may be in part related to slight monocytosis and in part to Th2 activation. IL-6, NO and CRP were unambiguously involved in the host parasite interaction and therefore may be useful markers in monitoring CE management and evaluating surgical stress. PMID- 16218216 TI - Leishmaniosis due to Leishmania infantum in a FIV and FelV positive cat with a squamous cell carcinoma diagnosed with histological, serological and isoenzymatic methods. AB - Leishmaniosis caused by Leishmania infantum is an endemic zoonosis present in the Mediterranean area. Canidae (dog and fox) constitute the main reservoir hosts for the parasite, whilst wild rodents or the cat can be carriers of the protozoan and are considered as secondary potential reservoirs. This paper describes a case of disseminated feline leishmaniosis with cutaneous (ulcerative), visceral (spleen and lymph nodes) and blood involvement in a FIV-FelV positive cat. The microscopic identification of the Leishmania infection was initially made on a skin biopsy of the temporal area, where a squamous cell carcinoma was diagnosed. The diagnosis of the disease was achieved by several serological techniques (ELISA, IFAT and Western-blot). The strain was obtained by blood culture, characterized by electrophoresis of isoenzymes and identified as Leishmania infantum zymodeme MON-1. Since the infection due to L. infantum is a zoonosis, the potential feline reservoir should be more investigated. Serological analysis by Western blot on domestic cats provides a useful tool. In veterinary practice, feline leishmaniosis should be systematically included in the differential diagnosis when compatible cutaneous lesions are present, especially in the endemic areas of canine leishmaniosis. PMID- 16218217 TI - Phoretic association of mites and mallophaga with the pigeon fly Pseudolynchia canariensis. AB - Myialges anchora Trouessart, 1906 and M. lophortyx (Furman & Tarshis, 1953) gravid females, surrounded by clusters of eggs, were found strongly inserted into the cuticle of head, thorax, abdomen, femurs and wings of Pseudolynchia canariensis (Macquart, 1840), a hippoboscid fly parasite of the pigeon. This lousefly results obligatory host for ovigerous females of Myialges and for the development of their eggs, and phoretic host because the dispersal of hatching larvae to new hosts may then occur with dispersal of fly carriers. Together with the Myiolges species, not ovigerous females of Ornithocheyletia hallae Smiley, 1970 and Columbicola columbae (Linnaeus, 1758) were found on the pigeon fly. PMID- 16218218 TI - [World news in age-related diabetes: blood sugar lowering can improve prognosis]. PMID- 16218220 TI - [Thus physicians succeed in transference of diet tips to their own body. Build in more enjoyment in your life!]. PMID- 16218219 TI - [Physicians and their overweight. When the physician's jacket is tight]. PMID- 16218221 TI - [Illegal drug trade discovered in Bavaria. How can we protect ourselves from drug falsification?]. PMID- 16218222 TI - [Tennis and golf arm. Elbow ache. So you certainly clarify]. PMID- 16218223 TI - [Questions on "Whole body sport kind with drive". Nordic walking after disk prolapse, too?]. PMID- 16218224 TI - [Those who want to get to the top need counseling. Who can tolerate the heights?]. PMID- 16218225 TI - [Hazards of mountain climbing and hiking]. AB - At elevations above 1500 m, even a healthy person undergoes acclimatization. To avoid problems such as acute mountain sickness (AMS), high altitude cerebral edema (HACE) or high altitude pulmonary edema (HAPE), the speed of ascent and the daily sleeping elevation are of primary importance. Mild symptoms and peripheral swelling are usually harmless. However, when the severity of altitude sickness progresses, rapid therapy and immediate transport to lower elevations can be life saving under certain conditions. A sojourn in the mountains requires effective preparation and prophylaxis against oxygen deficiency, increased UV radiation, as well as against the possibility of hypothermia and frostbite. PMID- 16218226 TI - [Planned sojourn at high elevations--what the primary care physician should know]. AB - For a healthy, active person, the chance that new ischemic events will occur up in the mountains is not any higher than in the valley. However, for inactive persons with health risks, there is a chance that the stress at the unaccustomed elevation could provoke a clinically silent coronary insufficiency. People with hypertension or diabetes should be able to monitor and adjust the dosage of their medication themselves. Asthmatic patients are often surprisingly fit even in the mountains. When the medication dosage is well established, they may also go mountain climbing at high elevations. On the other hand, when COPD or pulmonary hypertension is present, a pulmonologist should be consulted to determine whether high elevations could cause problems for the patient. Patients who have a medical history of thromboembolisms also require special preparation and prophylaxis. Migraine sufferers should acclimate themselves particularly conscientiously because migraine symptoms are very difficult to differentiate from that of acute mountain sickness. PMID- 16218227 TI - [European Hypertension Congress initiative: implement modern risk management without further delay!]. PMID- 16218228 TI - [Self-inflicted injurious behavior]. AB - Self-inflicted injurious behavior Auto-aggressive behavior such as the simulation of crimes, attempted insurance fraud or suicidal intent are observed in psychiatric diseases and, in particular, in borderline personality disorder. The self-inflicted injuries often show typical characteristics, the knowledge of which is helpful for making a diagnosis and can critically influence further therapy. Borderliner injuries are often noticeably uniform, arranged in parallel, superficial and on body locations that are easily reached. For affected persons who are at high risk for suicide, it is of great importance that they receive psychiatric therapy as soon as possible. PMID- 16218229 TI - [One change after the other. Thus is accounted for in emergency services today]. PMID- 16218230 TI - [Differential diagnosis of verrucous skin changes. When wild warts grow rapidly...]. AB - This article gives and overview of the various types of verrucous skin changes (viral warts, seborrheic warts). PMID- 16218231 TI - [Cutaneous horn on the lower lip]. PMID- 16218232 TI - [Education, roles, culture: a profession on the border]. PMID- 16218233 TI - [The theses in tri-annual degree courses for nurses: towards a better orientation of the final dissertations for the nursing degrees]. PMID- 16218234 TI - [Quality of life in transplanted patients]. PMID- 16218235 TI - [Explorative study on opinions, skills and perceived value of the dissertation of the nursing students of the degree course of Verona University]. PMID- 16218236 TI - [The role of the nurse in helping terminal cancer patients to fulfill their project in the last days of life]. PMID- 16218237 TI - [Variability in techniques related to prone positioning in ARDS patients]. PMID- 16218238 TI - [Parents' knowledge on SIDS prevention: the results of a survey]. PMID- 16218239 TI - [Competences and skills of expert nurses in the assessment of breathing problems: the results of a survey]. PMID- 16218240 TI - [The monitoring of surgical drainages: is there a "normal" standard?]. PMID- 16218241 TI - [Patient's problems in managing Kehr tube at home]. PMID- 16218242 TI - [The integration of hospital policies and educational needs: the description of an experience]. AB - The Educational Office of the Hospital and Health District of Venice offered the nurses completing their degree course in Padua University, a support for the final dissertation. The students were invited to select problems or issues relevant for the Hospital or District and were asked to make their final work available to the hospital and their colleagues. Twenty students were involved in this project. The support offered included the development of the study protocol and the bibliographic search. Four articles were published after the final dissertation and/or presented to local conferences. Two contributions were used to introduce relevant changes in the organization of the Health Department. Moreover, the Health Department can use the bibliographic searches, protocols, nursing guidelines and data produced to improve the training of nurses and the local organization of health services. PMID- 16218243 TI - [Factors that influence the duration of peripheral venous catheters used to antibiotic therapy in Cystic Fibrosis patients: results of a study]. AB - Antibiotic intravenous (IV) therapy is of paramount importance in the treatment of cystic fibrosis (CF) lung disease. During their life CF patients have to undergo frequent IV antibiotic treatments. Peripheral venous catheters (PVC) -or cannulas- are often used to reduce the number of venepunctures needed to complete a treatment cycle, as well as to provide patients the best possible level of autonomy. The clinical experience in managing IV therapy in CF patients at Meyer Hospital in Florence, suggested that the in situ duration of PVCs varies greatly. GOAL: To understand which factors influence the duration of PVCs used to administer IVantibiotic in people with CF. STUDY DESIGN: Cohort prospective study, integrated with retrospective data retrieval. MATERIALS, PATIENTS AND METHODS: We observed all the PVCs inserted during an 8-month period to CF patients followed at the CF Centre of Tuscany (Florence, Italy). RESULTS: We studied 87 PVCs inserted to 36 patients in 56 IV antibiotic treatment cycles. The mean duration of PVCs was 8.3 days. The mean duration of PVCs used for 3 daily antibiotic administrations was longer than that of PVCs used for 2 daily antibiotic administrations (10.3 days vs. 6.5 days, p=0.001). Moreover, PVCs with a larger gauge (G22) had longer mean duration than smaller (G24) PVCs (9.6 days vs. 7.2 days, p=0.03). No statistically significant association was found between PVCs' duration and, respectively, the place where the treatment was administered (in hospital or at home), the arm used (dominant or non-dominant), the PVC's brand, patient's clinical condition and the concurrent use of either steroidal antinflammatory drugs or NSAIDs per os. PMID- 16218245 TI - [Microcredit: the alternative option]. PMID- 16218244 TI - [The terminal heart failure patient]. PMID- 16218248 TI - State of Alabama Office of Women's Health "Alabama Together One Step at a Time". Combating obesity in Alabama women. PMID- 16218247 TI - Healthcare contracts, organizational responsibility and licensing. PMID- 16218249 TI - Angel Flight story. PMID- 16218250 TI - Nurses-defenders of environmental health, polyvinyl chloride (PVC). PMID- 16218252 TI - The elder orphans: who are they? PMID- 16218251 TI - Nurse fatigue: the human factor. PMID- 16218253 TI - The pocket guide to patient loyalty. PMID- 16218254 TI - Collaborative cancer care. PMID- 16218255 TI - Clinical predictors of survival in advanced cancer. AB - Prognostication is one of the three cardinal clinical skills. Although it has been undervalued by modern medicine compared with diagnostics and therapeutics, poor prognostication can have dire consequences for the patient with advanced cancer, almost as serious as the wrong diagnosis or treatment. Oncologists relying on their subjective judgment for predicting survival often will be inaccurate, usually as too optimistic, which may result in overly aggressive cancer treatment. Actuarial judgment, based on assessment of statistically derived key factors, has the potential to improve prognostic accuracy. These factors in patients with advanced cancer differ from those in patients with newly diagnosed disease; they include performance status, symptoms of the cancer cachexia syndrome, and patient-rated quality of life, rather than tumor size, tumor grade, or extent of disease. Laboratory markers such as leukocytosis or lymphopenia also appear to be useful. Novel markers include acute phase reactants (C-reactive protein, vitamin B12) and cytokines that may provide more objective evidence of survival prospects. Prognostic indices, nomograms, and web-based tools are in development for the advanced cancer population. Identifying the clinical markers predicting for short-term survival in patients with advanced cancer is important to help form the basis for teaching prognostication skills to physicians. PMID- 16218256 TI - The questions of prognosis. PMID- 16218257 TI - The clinical and research implications of survival prediction. PMID- 16218258 TI - Pathogenesis and management of cancer-related insomnia. AB - Insomnia is the most common sleep disorder reported by cancer patients. Oncologists have noticed that alterations in sleep patterns are endemic among their patients, yet sleep problems are rarely assessed in a typical patient evaluation. Other concerns, such as morbidity and mortality, appear to take precedence. The cause of chronic sleep difficulties is multifaceted and up until recently, little attention has been given to the potential factors associated with the pathogenesis of cancer-related insomnia. The unique contributions of psychologic, medical, treatment side effects, environmental, behavioral, and pharmaceutical pathways on cancer-related insomnia cannot be ignored. This paper explores an overview of the incidence and severity of sleep disturbance in cancer patients, a review of the mechanisms of sleep, and the potential factors associated with the pathogenesis of cancer-related insomnia. Nurses, physicians, and other healthcare providers are in a unique position to greatly improve the quality of sleep in cancer patients. Cancer patients face many challenges; sleep problems do not have to be one of the necessary consequences associated with the cancer experience. PMID- 16218259 TI - Are refined differential diagnosis and pharmacologic treatment for cancer-related insomnia always necessary? Practical recommendations for cancer care providers. PMID- 16218260 TI - Principles for managing cancer-related sleep disturbances. PMID- 16218261 TI - Palonosetron improves prevention of chemotherapy-induced nausea and vomiting in elderly patients. AB - Although elderly patients have been reported to be less prone to chemotherapy induced nausea and vomiting (CINV), its management is complicated by a high frequency of comorbidities and polypharmacy and an increased risk of dehydration and impaired cognition. The comparative efficacy and tolerability of palonosetron and ondansetron/dolasetron were assessed in a retrospective post hoc analysis using pooled data from 171 elderly patients (age > or = 65 years) with cancer enrolled in two randomized, double-blind, phase III clinical studies comparing single IV doses of these antiemetic agents given prior to receipt of moderately emetogenic chemotherapy. The complete response rate during the postchemotherapy period was significantly higher in the palonosetron group than in the ondansetron/dolasetron group in the 5 days following chemotherapy. The proportion of patients who were nausea-free on the problematic days 2 and 3 post chemotherapy and the time to treatment failure also significantly favored palonosetron. In this population that included patients with pre-existing comorbidities, palonosetron was well tolerated, with similar or fewer adverse events than the comparators. Comparisons in electrocardiogram parameters revealed that the mean postdose change from baseline in QTc interval was 3 ms for palonosetron 0.25 mg and 5 ms for ondansetron/dolasetron. In this retrospective analysis, palonosetron provided superior efficacy to ondansetron/dolasetron for the treatment of CINV in elderly patients receiving moderately emetogenic chemotherapy. Based on its safety profile, antiemetic control, and convenient dosing, palonosetron can be recommended for use in elderly patients with cancer receiving emetogenic chemotherapy. PMID- 16218262 TI - The Functional Assessment of Anorexia/Cachexia Therapy (FAACT) Appetite Scale in veteran cancer patients. AB - Anorexia and appetite assessment is an important priority in supportive oncology. A series of 156 veterans participating in a hematology oncology service completed the Functional Assessment of Anorexia/Cachexia Therapy (FAACT), the Functional Assessment of Cancer Therapy-General scale, the Memorial Symptom Assessment Scale Short Form (MSAS-SF), and the Zung Self-Rating Depression Scale and were followed for survival. The FAACT score correlated well with Karnofsky performance status, quality of life, and symptom distress subscales. A single appetite distress item from the MSAS-SF correlated well with these measures. Both appetite measures correlated with the presence of other symptoms and with concurrently measured hemoglobin, serum sodium, albumin, and cholesterol levels. These self-reported appetite measures were univariate predictors of survival and contributed additional prognostic information to data related to weight-loss distress. In a smaller study, the FAACT score correlated with a visual analogue measure of appetite and with the North Center Cancer Treatment Group appetite instrument. These data support use of these tools for the evaluation of appetite concerns among patients with advanced cancer. PMID- 16218264 TI - [Diabetic embryopathy. Report of hundred cases]. PMID- 16218263 TI - How to reduce fears of legal/regulatory scrutiny in managing pain in cancer patients. PMID- 16218265 TI - [Treatment of congenital insufficiency in fibrin stabilising factor (FSF or factor XIII) by purified factor XIII of placental origin. Report of two cases with a four year follow up]. PMID- 16218267 TI - [Primary fibroelastosis of the endocardium. Genetic advice]. PMID- 16218266 TI - [Smith-Lemli-Opitz dwarfism. Report of four cases. Review of the literature]. PMID- 16218268 TI - [The Zollinger-Ellison syndrome in child. Report of one case and review of the literature]. PMID- 16218269 TI - [Neonatal gastric necrosis. Two new cases including one with a gastric capillary hemangioma]. PMID- 16218271 TI - [Recurrent pneumopathy by abnormal implantation of the bronchi of the right upper lobe. Report of two cases]. PMID- 16218270 TI - [Spontaneous pneumothorax and excaved pulmonary metastases in a child with a mesenchymatous tumour]. PMID- 16218272 TI - Bolstering bottom lines. Claims and coding. PMID- 16218273 TI - Claims & coding HotList. PMID- 16218274 TI - Like it? Yes. Need It? Yes. Buy it? Nah. PMID- 16218275 TI - Capacity management: the bedrock of efficiency. PMID- 16218276 TI - Securing the healthcare border. PMID- 16218277 TI - Streamlining pediatric patient care. PMID- 16218278 TI - Saving lives with teletrauma video communications. PMID- 16218280 TI - Right from the start. PMID- 16218281 TI - Patient privacy: the right to know versus the need to access. PMID- 16218282 TI - Ten hard questions about the future of the specialty. PMID- 16218283 TI - Taking care of children. PMID- 16218284 TI - Taking care of children. PMID- 16218285 TI - Is doing good good enough? PMID- 16218286 TI - Is doing good good enough? PMID- 16218287 TI - Is doing good good enough? PMID- 16218288 TI - Diary missed already. PMID- 16218289 TI - How to change behavior. PMID- 16218290 TI - Helping patients help themselves. PMID- 16218291 TI - Terminating a patient: is it time to part ways? PMID- 16218292 TI - Make medical notes better and faster with macros. PMID- 16218293 TI - ICD-9 changes: it's time to tend to your superbill. PMID- 16218294 TI - Coding "routine" office visits: 99213 or 99214? PMID- 16218295 TI - Meet Dr. Wonderful. PMID- 16218296 TI - HIT adoption offers big savings, but logistics worries persist. PMID- 16218297 TI - Smaller physician practices sluggish in EHR adoption. PMID- 16218298 TI - Low comprehension of consent forms creates surge in medical errors. PMID- 16218299 TI - Pay-for-performance incentives: are you missing out on revenue? PMID- 16218300 TI - Patient safety legislation removes reporting barriers. PMID- 16218301 TI - Patient safety alert. HCA nears completion of systemwide eMAR. PMID- 16218302 TI - Discharge planning advisor. Innovative program helps homeless, frees up beds. PMID- 16218303 TI - Quality leaders work to reduce surgical infections. PMID- 16218304 TI - Did we really make a difference? PMID- 16218305 TI - Kudos for tobacco issue. PMID- 16218306 TI - All options on table in wake of Supreme Court decision. PMID- 16218307 TI - Do prophylactic anticonvulsants in patients with brain tumors decrease the incidence of seizures? AB - The well-done systematic review included a moderate number of patients from randomized controlled trials with an objective diagnosis of a brain tumor. There were mild differences between the 5 studies used in the systematic review with patients in 3 of the studies undergoing surgical resection or debulking of their disease. There was no evidence of seizure prevention overall or in the subgroup analysis. Given the small number of patients in each subgroup, these results should be interpreted with caution. Known adverse effects with anticonvulsant therapy include severe rash (including Stevens-Johnson syndrome), hematologic effects, and drug-drug interactions. Therefore, we should be hesitant to place patients on these medications without a proven benefit. The relatively small number of patients in this meta-analysis means that larger scale studies could show a small clinical benefit from anticonvulsants. PMID- 16218308 TI - The Badger Heart Program: a successful model for improving health literacy and helping women make lifestyle changes. PMID- 16218309 TI - Genetic services plan for Wisconsin. AB - During 1999-2001, a multidisciplinary group met to develop a plan for the future of genetic services in Wisconsin. The result was The Genetic Services Plan for Wisconsin (available at www.slh.wisc.edu/genetics/stateplan_toc.html), a problem oriented needs-identification guide to address current and future challenges likely to affect the provision of genetic services in Wisconsin. The Plan is directed to all individuals who have a stake in the future of medical genetic services in Wisconsin. These include, but are not limited to, primary and other health care professionals, genetics professionals, governmental representatives, policy makers, legislators, educators, third-party payers, and current and potential consumers. This article provides an overview of The Genetic Services Plan for Wisconsin (Plan) and highlights the recommendations made for the continuing integration of new genetic knowledge across the continuum of medical care delivery in Wisconsin. PMID- 16218310 TI - Is it time to regulate the pharmaceutical industry? PMID- 16218312 TI - Badger Heart Program: health screenings targeted to increase cardiovascular awareness in women at four northern sites in Wisconsin. AB - PURPOSE: This study reports the results of initial statewide health screenings of the Badger Heart Program (BHP), which aims to increase the cardiovascular health awareness and health status of women living in Wisconsin. METHODS: In May 2004, the BHP provided cardiovascular disease risk screenings to women in 4 geographic areas throughout Wisconsin. Screening participants were informed of their results and counseled on issues concerning diet, exercise, smoking cessation, medications, and/or visiting their primary health care professional. Data collected included total cholesterol, high-density lipoproteins, low-density lipoproteins (LDL), triglycerides, blood pressure (BP), blood glucose measurements, height and weight, along with a brief survey of medical history, family history, smoking status, and current medication use. After the screening, participants were asked to volunteer for an opt-in 6-month educational program. RESULTS: A total of 318 women participated. The majority screened were Caucasian (95.9%) with an average age of 58 years (standard deviation [SD] = 13.2). Participants' blood pressure and LDL goals were determined from the risk profile assessment. According to national guidelines, an optimal blood pressure goal of < 140/90 was recommended for the majority of the participants (294 [92.5%]), while an optimal blood pressure goal of < 120/80 was recommended for only 24 (7.5%) participants. An optimal LDL value <100 was recommended for 48 (15.4%) participants, an LDL goal of <130 was recommended for 106 (33.3%), and an LDL goal of less than <160 was recommended for 157 (50.5%) participants. Of screened participants, 35% were not at BP goal, 32.4% were not at LDL goal, and 53.5% were not at both goals. CONCLUSION: While the number of participants who were at BP and/or LDL goal is higher than what is generally reported in the literature, there is still opportunity for significant improvement. A follow-up analysis including re-screening of individuals aimed at measuring the improvements in CVD profile post educational interventions will occur in November. PMID- 16218311 TI - Vacations improve mental health among rural women: the Wisconsin Rural Women's Health Study. AB - OBJECTIVE: To compare psychological stress, quality of marital life, and disruptive homelife due to work among rural women of central Wisconsin who take vacations frequently and those who do not. METHODS: Women were recruited from 1996 to 2001 for a prospective cohort study from the Marshfield Epidemiologic Study area, a geographic area in central Wisconsin. Stratified sampling was used to select a random sample of 1500 farm and non-farm resident women. RESULTS: The odds of depression and tension were higher among women who took vacations only once in 2 years (Depression: OR=1.92, 95% CI=1.2,3.0; Tension: OR=1.7, 95%CI=1.2, 2.3) or once in 6 years (Depression: OR=1.97, 95% CI=1.2, 3.2; Tension: OR=1.9, 95% CI=1.3,2.8) compared to women who took vacations twice or more per year. The odds of marital satisfaction decreased as the frequency of vacations decreased. CONCLUSION: Women who take vacations frequently are less likely to become tense, depressed, or tired, and are more satisfied with their marriage. These personal psychological benefits that lead to increased quality of life may also lead to improved work performance. PMID- 16218313 TI - Outcomes of complex gastrointestinal procedures performed in a community hospital. AB - BACKGROUND: Complex gastrointestinal (GI) procedures have been defined as those that are associated with higher morbidity and mortality, require a high level of technical expertise, and occur in less than 6000 patients per year in the United States. Prior studies suggest a direct volume-outcome relationship. HYPOTHESIS: Complex GI procedures may be performed with good outcomes in a lower volume hospital with a commitment to surgical residency training. METHODS: Retrospective chart review of all patients undergoing non-emergent operations that are considered complex GI procedures (esophagectomy, total gastrectomy, major hepatic resection, pancreaticoduodenectomy, biliary tract anastomosis, and total abdominal proctocolectomy) from July 1989-June 1997 in a rural referral medical center. RESULTS: One hundred six consecutive patients underwent complex GI procedures during a 7-year period ending June 1997. Patients ranged from 19-90 years (mean 62). Forty-eight patients (45.3%) had 1 or more major comorbidities. Seventy-three patients (68.9%) had operations for malignancies. Average length of stay (LOS) was 13.2 days (range 5-38). Major complications occurred in 15 patients (14%). Two patients died (mortality 1.9%), 1 after esophagectomy and 1 after a Whipple procedure. LOS, morbidity, and mortality were less than or equivalent to published reports from high-volume medical centers. CONCLUSION: Excellent outcomes for complex GI procedures can be achieved at lower volume medical centers. Regionalization strategies to improve patient care should be based on outcome studies rather than volume alone. PMID- 16218314 TI - Deep brain stimulation of the subthalamic nucleus in Parkinson's disease. AB - OBJECTIVE: To evaluate the clinical effects of subthalamic nucleus deep brain stimulation in patients with Parkinson's disease within the first 12 months after surgery. METHODS: We performed a prospective study in 8 patients with Parkinson's disease, in whom electrodes were implanted in the subthalamic nucleus bilaterally. We compared levodopa-equivalents and the scores of the Unified Parkinson's Disease Rating Scale pre- and post-operatively. The post-operative evaluation was done between 3 and 12 months after surgery. RESULTS: Antiparkinsonian medications were reduced post-operatively by a mean of 61.5% (P < 0.01) from a levodopa-equivalent dosage of 1144.9 +/- 572.5 mg/day to 440.9 +/- 172.1 mg/day. Motor scores improved 44.4% (P < 0.01) and activities of daily living scores 38.2% (P < 0.01). Adverse events included a subcutaneous hematoma in 1 patient after internal pulse generator implantation necessitating evacuation. CONCLUSIONS: Bilateral stimulation of the subthalamic nucleus is associated with significant improvement in motor function and reduction of antiparkinsonian medications in patients with Parkinson's disease in the first 12 months after surgery. On-state dyskinesias were greatly reduced, probably due to the reduction of total antiparkinsonian medications. The procedure is well tolerated. PMID- 16218315 TI - Intradiscal electrothermal annuloplasty therapy: a case series study leading to new considerations. AB - OBJECTIVE: To evaluate outcomes of intradiscal electrothermal annuloplasty (IDEA) therapy in the treatment of chronic discogenic low back pain in consecutive IDEA patients treated at a rural pain management clinic. STUDY DESIGN: An observational case series study design was applied to consecutive IDEA patients qualifying under the inclusion and exclusion criteria. Patient assessment of pain and disability were performed at baseline and 6 weeks, 3, 6, 12, and 24 months post-IDEA. METHODS: Selected patients underwent IDEA for an average of 15 minutes at a temperature of 90 degrees C. Analyses of outcomes included Visual Analog Scale (VAS) assessments of levels of pain, and Roland Morris Disability Questionnaire (RMDQ) assessments of functional capacity at pretreatment, and 6 weeks, 3, 6, 12, and 18 months post-treatment time points. RESULTS: At 6 months post-IDEA treatment, patients (n=51) demonstrated statistically significant improvement (P < 0.001) as measured by a mean change of over 20 points from the pretreatment score on the RMDQ. At 1 year, post data remained significant in the 33 patients who had achieved this time point. VAS pain data were also statistically significant at 6 months (P = 0.023). Analysis of patient profiles revealed that statistically significant improvement of pain and functional capacity was strongly associated with female gender and age (range of 18-45 years), and that statistically significant improvement was not sustained in males beyond the 3-6 month point. These data support the outcomes reported in the few existing observational studies to date. Of 86 patients receiving IDEA therapy, 73 provided RMDQ data at baseline and at 3 months or later and were included in the analyses. Some patients were lost to follow-up at later time points. CONCLUSIONS: These data show favorable outcomes after IDEA therapy, and suggest that women may experience more improvement than men, particularly with regard to perceived disability improvements. Data suggest that greater improvement in IDEA outcomes may be achieved by profiling the characteristics of patients who achieve the optimal long-term outcomes following treatment and should be considered during evaluation of patient eligibility for IDEA. PMID- 16218316 TI - Dietary calcium intake among women attending primary care clinics in Wisconsin. AB - BACKGROUND: Osteoporosis and related fractures are becoming more prevalent as the population ages. Primary care professionals counsel all women about adequate calcium intake as part of an overall osteoporosis preventive strategy. Population data shows that the average calcium intakes in women are substantially lower than standard recommendations. No data is available on dietary calcium intakes among a primary care population. This study looks at dietary calcium intakes and calcium supplementation among a sample of women attending 4 primary care clinics in Wisconsin. METHODS: A total of 210 women completed food frequency questionnaires while waiting to see a health care professional in the waiting room of 4 primary care clinics. The women estimated amount of high calcium foods eaten per day and frequency of ingestion per week. Women also indicated whether or not they were taking a calcium supplement. RESULTS: The overall mean calcium intake was 1309 mg per day. Women who were nonsmokers, postmenopausal, older, Caucasian, or who had been diagnosed with osteopenia or osteoporosis had higher calcium intakes. Over half of the study population took a calcium supplement regularly. CONCLUSION: Women in a primary care population in Wisconsin had higher dietary calcium intakes than women from population studies. Primary care professionals need to continue counseling women about adequate calcium and vitamin D intakes as part of an overall osteoporosis prevention program. PMID- 16218317 TI - Optimal collection technique and devices for a quality pap smear. AB - BACKGROUND: Collection technique is critical in the performance of the Papanicolaou (Pap) smear as an important screening tool for cervical cancer. While superior devices have been established, less effective devices continue to be used for both conventional and liquid-based Pap smears. Our aim is to determine the performance of collection devices currently used in obtaining conventional Pap smears and whether sequence of collection is important for higher quality results. METHODS: This prospective, blinded, cohort study used 2 cytology labs to analyze Pap smears done within a 1-year period. Study participants were 128 clinicians who practice in Dane County, Wis, who send their Pap smears to either of the 2 study cytology labs. Participants included advance practice nurses, family physicians, and obstetrician/gynecologists. Logistic regression was utilized for analysis. RESULTS: In conventional Pap smears, sequence of collection did not affect any quality indicators. The Cervex-brush (broom) was associated with absent endocervical cells (Odds Ratio = 3.12, P < .001), limited or unsatisfactory results (OR = 1.68, P < .01), and obscuring inflammation (OR = 2.01, P < .01). Of those clinicians who had high levels of absent endocervical cells on their Pap smears (defined as > 3/30 Pap smears), 47% used the broom alone. The Cytobrush optimized quality indicators, and the combination of the Cytobrush for the endocervix and spatula for the ectocervix was superior. Presence of infectious agents also contributed to the absence of endocervical cells (OR = 3.09, P < .001). CONCLUSIONS: The combination of the Cytobrush (endocervix) and spatula (ectocervix) is superior for a quality Pap smear. The sequence of collection was not important in conventional Pap smears. The broom alone performs poorly. Presence of infection decreases quality. PMID- 16218319 TI - Gene-based drug prescribing: clinical implications of the cytochrome P450 genes. AB - The Institute of Medicine recently mandated an increased effort to improve patient safety and reduce medical error. With the description of genetic polymorphisms in the drug metabolizing enzymes, the field of pharmacogenetics may improve medical care through a reduction in both therapeutic failure and adverse drug reaction. Investigators at the Marshfield Clinic in central Wisconsin are piloting the process of gene-based drug prescribing in a variety of contexts. This paper reviews the field of cytochrome P450 (CYP) genetics and explores factors that impact the utility of this information in clinical practice. PMID- 16218318 TI - Central pontine myelinolysis: case series and review. AB - OBJECTIVE: To review central pontine myelinolysis (CPM) and osmotic demyelination syndrome (ODS) and describe the clinical features, etiologic factors, and clinical outcomes of 6 patients diagnosed with CPM or ODS. STUDY DESIGN: A retrospective case series. METHODS: Medical records of patients diagnosed with CPM or ODS at Marshfield Clinic/St. Joseph's Hospital from 1986 to 2003 were reviewed. Chart abstraction was completed with a standardized data abstraction form. RESULTS: Six patients were identified, ranging in age from 31-73 years (mean age = 51.5 years). Clinical presentations were nonspecific. Common symptoms included lethargy and dysarthria. Five of the 6 patients had chronic alcoholism. All had improvements in their clinical conditions upon hospital discharge. CONCLUSIONS: CPM and ODS are rare demyelinating diseases of the pons and extrapontine sites. Prompt diagnosis and management of associated complications are essential for favorable clinical outcomes. PMID- 16218320 TI - Compartment syndrome and fatal rhabdomyolysis in sickle cell trait. AB - Sickle cell trait is a relatively common condition in the African-American population. Individuals with this condition may have any of several complications under rare circumstances. We report a patient who presented with extensive compartment syndrome leading to death. A 31-year-old African-American male with known history of sickle cell trait developed extensive compartment syndrome followed by rhabdomyolysis, severe acidosis, acute renal failure, and coagulopathy. Although the patient underwent multiple fasciotomies for extensive compartment syndrome and received aggressive resuscitation with massive transfusions of blood, blood products and intravenous fluids, he died. Multiple compartment syndromes in patients with sickle cell trait represent a very unusual complication and can cause significant morbidity and mortality. Avoidance of strenuous exercise, especially in hot climates, early diagnosis, and aggressive therapy are very important to prevent this catastrophic complication. PMID- 16218321 TI - Plan to address physician shortage requires proper support. PMID- 16218322 TI - Your retirement nest egg: are you creating a reliable stream of income? PMID- 16218323 TI - MetaStar's rural health projects. PMID- 16218324 TI - Noise and health. PMID- 16218325 TI - Center for Occupational Health at the National Institute of Public Health. WHO Collaborating Center for Occupational Health. PMID- 16218326 TI - Quality of Life, Sustainable Future and medical education. AB - Substantial social and environmental changes in a global world as well as a new paradigm of medicine emphasizing high technology and evidence based approach bring to the current medicine many paradoxes. Undoubtedly, new era creates many positive opportunities and challenges for medical professionals. On the other side, traditional concept of medicine as basically humanistic "helping profession" is rather threatened. In this context, there is a need for a re defining of medical curricula and to offer to the medical students the concepts and ideas which reflect a current development. Traditional public health terms such as "Primary Prevention", "Health Promotion", "Health Protection", "Health Determinants" should be re-defined and the new ones such as "Quality of Life" and "Sustainable Future" must be introduced as an integral part of medical education. The relevant concepts are discussed in the context of specific situation of the health care transformation in Central and Eastern European Countries. PMID- 16218327 TI - Factors determining changes in self-rated health in the Polish community-dwelling elderly. AB - Changes in self-rated health and its determinants have been analyzed in the group of 551 community-dwelling older age citizens of Krakow during the 12-year interval observation. Multidimensional model showed that changes in self-rated health between the studies have been significantly determined by such variables like age and self-evaluation of health status in the Ist study. Self-rated health was also less markedly decreased in men, who continued professional activity in the Ist study. Among variables analyzed in the IInd study it was age-related functional disability reported by men and reported chronic conditions that deteriorated self-rated health significantly. Changes in self-rated health between the Ist and the IInd study among women were determined by the same variables as in men (except for the continuation of professional activities in the Ist study). Significantly diminished scores were found in women with higher level of functional activity in the Ist study and greater independence in performing daily activities in the IInd study. Analysis of the summary effect of chronic diseases on self-rated health has shown significantly greater deterioration of self-rated health between the first and the second study related to the number of diseases reported in the Ist study. PMID- 16218328 TI - Could socio-economic transformation and the resulting psychological stress influence cancer risk in Opole province, Poland? AB - The paper presents the results of a risk assessment analysis of cancer morbidity in Opole province before and after a political transformation in Poland, i.e. in the 1985-1989 quinquennium and the following two equivalent periods of: 1990-1994 and 1995-1999. Measures of morbidity are given and its growth in males and females are compared with the ageing effect as well as with unemployment. In the paper a general conclusion has been drawn suggesting that the socio-economic transformation begun after 1989 and the resulting stress could have been the one of the possible background effects influencing the health status in the region. It must be accentuated, however, that the relation has not been a subject of statistical proving due to a methodological impossibility; a divagated question is offered for scientific concern and open discussion. PMID- 16218329 TI - The role of physical activity in the prevention of stroke. AB - Although large-scale epidemiologic studies, as well as interventional trials have provided strong evidence of a consistent and robust association between physical activity and cardiovascular disease, the effect of exercise on the burden of stroke is not well understood and appreciated. This review has a purpose to summarize the literature on the effect of physical activity on stroke morbidity and mortality, and to provide current scientific evidences. PMID- 16218330 TI - Risk of congenital anomalies in the vicinity of waste landfills in Denmark; an epidemiological study using GIS. AB - Waste landfills are a potential hazard to health. Public concern exists about this potential hazard and researchers agree that further research is required on this field. The objective of the study was to investigate the association between waste landfill location and congenital anomalies risk in Denmark. The study was a multisided epidemiological geographical comparison study of risk of congenital anomalies combined and congenital anomalies of the cardiovascular and nervous systems with maternal residence in the vicinity of 48 Danish waste landfills compared with those living further away in the years 1997 to 2001. We used routine health and population data in Geographical Information System (GIS) to investigate the risk. The subjects were 2,477 live birth with congenital anomalies. All relative risks in the proximal zones of 0-2 km were set to 1 for comparison. For all anomalies combined relative risk in the middle zones of 2-4 km joint was 0.991 and in the distal zones of 4-6 km joint the relative risk was 1.013. For congenital anomalies of the nervous system, the relative risk in the middle zones was 1.226 and in the distal zones 1.113. For congenital anomalies of the cardiovascular system, the relative risk in the middle zones was 0.926 and in the distal zones 0.854. This result was not supported by the aggregated risk ratio mean. We found no association between waste landfill location and congenital anomalies combined or of the nervous system. However, we found small excess risk for congenital anomalies of the cardiovascular system. No causal mechanisms are available to explain these findings, but alternative explanations include approximated birth rates and residual confounding. It is our recommendation that more comprehensive multisided studies will be executed to examine the safety of waste landfills. PMID- 16218331 TI - Maternal employment status and isolated orofacial clefts in Hungary. AB - AIMS: To study the role of maternal employment status as indicator of socioeconomic status in the origin of isolated orofacial clefts (OFC) and in the use of periconceptional folic acid/multivitamin supplementation. METHODS: 1,975 cases with OFC (1,374 cases with cleft lip +/- palate and 601 cases with posterior cleft palate), 38,151 population controls without any defects and 20,868 patient controls with other isolated defects were compared in the population-based data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities (HCCSCA), 1980-1996. RESULTS: The proportion of professionals and managerials was lower, while the proportion of unskilled workers, housewives and others was higher in the mothers of cases with OFC compared with the population control group. However, the comparison of OFC and patient control groups did not show any difference in the employment status of mothers. A lower level of folic acid supplementation occurred in the professional and skilled worker mothers of cases with OFC compared with the population control group. This difference was confirmed by the comparison of folic acid used by mothers of cases with OFC compared with patient controls. An infrequent multivitamin use was displayed in the studied groups. CONCLUSIONS: The prevalence of OFC at birth shows a slightly lower maternal employment status as indicator of socioeconomic status than in the population control group. The higher level of maternal education does not imply a higher rate of folic acid supplementation in the group of OFC. PMID- 16218332 TI - Cadmium in urine of children and adults from industrial areas. AB - Various human activities result in the release of significant quantity of cadmium to the environment. The study population included adults (230) and children (100) residing near the tobacco plant and a control group, unexposed to elevated cadmium concentrations, living in unpolluted area in the same city. Biomonitoring methods were used to assess the effects of contaminated air on the health of the public (urine samples). Ambient air samples were collected near the cadmium related plant and in unpolluted area during the last ten years. Values of cadmium concentrations in urine found in polluted area were in range of 0.05-0.42 microg/g creatinine for children and 0.15-1.96 microg/g creatinine for adults. The results indicated statistical significant age-dependent differences in urine cadmium concentration between polluted and unpolluted area for adults. Boys and men had significantly higher urine cadmium levels than girls and women, but in children there were no significant gender-dependent differences in cadmium excretion found out. In conclusion, the results from the present study indicate that cadmium urine concentration in polluted area are very high, particularly in children. PMID- 16218333 TI - Some trace elements in coal of the Czech Republic, environment and health protection implications. AB - Mining for coal and its utilization have various impacts on the surrounding environment. Huge volumes of waste materials which are by-products of both the underground and open cast coal mining, pose one of the major environmental hazards in addition to air pollution caused by coal burning in power plants in the Czech Republic. Some of these risks could be reduced when having accurate and comprehensive data on coal quality. Statistical data processing of almost 35,000 coal samples from Late Paleozoic and Tertiary coal basins of the Czech Republic provided a unique information on the quality of lignite, sub-bituminous and bituminous coals and anthracites including the content of toxic trace elements (As, Be, Hg, Pb and Se). In this context related environment and health risks and protection implications are discussed. PMID- 16218335 TI - [Lipopolysaccharides of gram-negative bacteria as exogenous pathogens. Bacterial translocation in vivo, inflammation, and pathology of the cardiovascular system]. PMID- 16218334 TI - Peracetic acid and sodium hypochlorite effectiveness in reducing resistant stages of microorganisms. AB - A comparative study on the efficacy of peracetic acid and sodium hypochlorite in inactivating resistant stages of microorganisms such as Cryptosporidium, Giardia and Cl. perfingens was carried out. Furthermore the evaluation of the potential reciprocal correlation among the concentrations of the organisms was performed. The results obtained indicate that, at the same experimental conditions, peracetic acid and sodium hypochlorite have nearly similar reduction power against the resistant stages of Giardia and Cl. perfringens. Both the oxidants are instead less efficient in the abatement of Cryptosporidium oocysts. Findings have also confirmed our previous studies on the absence of association between Clostridium and the protozoa. PMID- 16218336 TI - [Volatile fatty acids in blood and saliva of children with gastroduodenal diseases]. PMID- 16218337 TI - [Introduction to the analytical cytochemistry of acute leukemia (a lecture)]. PMID- 16218338 TI - [Laser DNA-flow cytofluorometry in the differential diagnosis and prognosis of the cervix uteri cancer]. AB - Laser DNA-flow cytofluorometry (DNA-FC) was used to examine 102 primary patients with morphologically verified pathology of the epithelium of the cervix uteri. There was a significant reduction in the count of cells in the cellular cycle phase (CCP) G0/1 and its increase in the CCP S and G2 + M, as well as a rose in the cell proliferation index (PI) in actually invasive (n=45) and microinvasive cancer of the cervix uteri (CCU) (n=21) as compared with the baseline values (n=8), CIN I-II (n=7), and CIN III (21 patients with severe dysplasia and cancer in situ; their DNA-FC parameters were close and significantly indistinguishable). With unfavorable clinical and morphological factors of CCU prognosis (age over 50 years, postmenopause, stages II-III, high-grade tumor, more than 3 mm stromal invasion, more than 4 cm in size, lymphovascular invasion in the regional lymph nodes, the endophytic form of growth, and a primary focus in the endocervix), aneuploid tumors were more significantly frequently found. The poor CCU prognostic factors significantly decreasing total and relapse-free 4-year survival included tumors of aneuploid type, those containing greater than 40% of aneuploid cells, those containing less than 70% of tumor cells in CCP G0/1, those having 10% of tumor cells or more in CCP S or more than 30% cell PI. While predicting CCU, the DNA index (DNAI) and PI with 0.09 and 0.05 informative value coefficients, respectively are the most significant DNA-FC parameters (after Shenon). By using them, the risk of progressive disease may be predicted with 67.9% probability. When DNAI and PI are used in combination with the most significant clinical and morphological factors of CCU, the likelihood of the latter increases up to 89.3%. PMID- 16218339 TI - [Comparision of cytological study results and data of bio-molecular assay hybrid capture II in patients with precancer of the cervix uteri]. PMID- 16218340 TI - [Characteristics of the statistical data processing in the computed ploidometric diagnosis of melanocytic tumors of the skin]. PMID- 16218341 TI - [Hematological parallels in chronic viral hepatitis B and C]. PMID- 16218342 TI - [Current view of the role of neutrophils in antitumor immunity (a review of literature)]. PMID- 16218343 TI - [Some aspects of the diagnosis of opportunistic bacterial vaginal infections during pregnancy]. PMID- 16218344 TI - [Contemporary trends in the thoracic surgery view to the history, perspective]. AB - The development of thoracic surgery in Czechoslovakia is very closely connected with the 1st Medical Faculty of the Charles University in Prague. The first Czech surgical clinic was established in Prague at this medical faculty in 1882. Professor Vilem Weiss was the first Head of the clinic. The Second Surgical Clinic in Prague was established in 1922 and Docent, later Professor, Academic Jiri Divis was the second head of this clinic. Jiri Divis is rightfully considered the founder of the Czechoslovak School of Thoracic Surgery. Whereas pulmonary tuberculosis was the main problem thoracic surgery faced in the first half of the 20th century, pulmonary carcinoma is the main issue addressed in thoracic surgery in the second half of the 20th century and currently. What new treatments can we currently offer our patients? Bronchoplastic procedures (so called sleeve resections) have developed and carcinomas of the tracheobronchial tree-carina- can be treated in a similar manner. Development in the technology has brought staplers--machines for surgical suture. Staplers allow performing volume reductive pulmonary resections as one of the treatment options for advanced pulmonary emphysema. Introduction of the pulmonary transplantation method in practice on 22.12.1997, when Professor Pafko and his team performed the first pulmonary transplantation in Czech republic, was a major breakthrough in pulmonary surgery. The minimal invasive technique for thoracic surgery has developed further. Video-assisted thoracoscopy is now an integral part of thoracic surgery. Introduction of PET-CT examination brought great benefit to a wide range of medical disciplines, including thoracic surgery. Where may the future development in surgery and thoracic surgery take us? We are finding that specialisation is vital in the development of all surgical disciplines. In certain borderline or technically complex operations, thoracic and mediastinal surgery requires cooperation with cardiac surgeons. Thoracic diseases should therefore be centralised in centres with sufficient experience and resources for further development of the discipline. PMID- 16218345 TI - [Intracranial meningiomas--criteria for selection of the optimum therapeutic modality]. AB - The authors present their own proposal for a standard diagnostic procedure algorithm in intracranial meningiomas, which they used in their prospective, non randomized longitudinal study in a group of 30 subjects within a year. The following four criteria were assessed in each patient: age, physical condition according the ASA classification, location of the meningioma on the MRI (superficial, the scull base) and the growth invasivity using selective DSA (ACI+ACE) and MRI (the vascularization type, the oedema index and the change in the oedema signal intensity in 3.5 hours). The criteria helped to establish the optimum therapeutic procedure for each patient: embolisation without a follow-up surgery (2 subjects), observation (2 subjects), pre-operative embolisation (5 subjects) and surgery without preceeding embolisation (21 subjects). This study did not assess the intracranial meningiomas treatment outcome. It highlights significance of the diagnostic procedures standardization in order to establish their optimum therapeutic modality. PMID- 16218346 TI - [Surgery of ventral hernia using fascia lata graft]. AB - Severe intraabdominal sepsis sometimes recquires very aggressive surgery with repeated surgical revisions of abdominal cavity. This often leads to large ventral hernia. The authors present one of the possibilities of surgical management of the hernia using fascia lata graft. This method is easy to perform and cost effective because of autologous material, which is most ideal from the point of biological view. PMID- 16218347 TI - [Laparoscopic surgical procedures in children with the Hirschprung's disease- initial experience]. AB - Laparoscopically assisted resection of the large intestine in the Hirschprung's disease patients was introduced into practice worldwide in mid 90-ies of the last century. In the Czech Republic this procedure had not been conducted. The aim of this work was to introduce the method in the Czech Republic. In this study the authors present their initial experience with this new method. They have successfully operated three patients with this disorder. PMID- 16218348 TI - Initial experience with the use of fibrin sealant for the fixation of the prosthetic mesh in laparoscopic transabdominal preperitoneal hernia repair. AB - INTRODUCTION: Laparoscopic inguinal hernia repair offers more rapid recovery and less pain than with the traditional open approach. However, injury to the nerves of the lumbar plexus with subsequent chronic pain or neuralgia has a reported incidence of 2% during laparoscopic hernia repair, particularly when the transabdominal preperitoneal technique (TAPP) is used. These complications are inherent to the use of staples for fixation of the mesh. To avoid nerve irritation, we considered the use of fibrin sealant for the fixation of the mesh instead of staples. The aim of this study was to evaluate this technique and to compare the short-term follow-up of these patients with patients who underwent the staple repair technique. This is the first reported use of fibrin sealant in laparoscopic TAPP hernia repair. METHOD: Between September and November 2004, we performed 17 consecutive laparoscopic hernia repairs (TAPP) in 14 patients (3 bilateral hernias) with primary hernias. The prosthetic mesh was fixed (10 x 15 cm) with 1 ml fibrin. The fibrin was applied using a special laparoscopic applicator. The peritoneum was closed with absorbable sutures. The postoperative course of these patients was compared with a cohort of matched patients who received the traditional staple fixation of the prosthetic mesh. RESULTS: Patients were evaluated at a median follow-up of 10.4 months (3.8-16.0 months). All patients underwent postoperative physical examinations. No recurrent hernia was found. There were 2 seromas and one hematoma in the stapled group. In the stapled group, one patient had pain in the area of the lateral femoral cutaneous nerve. There was no postoperative complication in the non-stapled group. CONCLUSION: Fibrin fixation of the mesh during laparoscopic transabdominal preperitoneal inguinal hernia repair is feasible without higher risk of recurrences. In addition the fibrin fixation method may decrease postoperative neuralgia and reduce the incidence of postoperative seromas and hematomas. PMID- 16218349 TI - [Exenteration procedures in the pelvis]. AB - Exenteration pelvic procedures are surgical options for treatment of locally advanced pelvic tumors. Due to the procedure's success rates, it has become a standard therapeutic procedure, when indicated. From the medical point of view, the following factors characterize the level of seriousness of these procedures: the fact that the procedure is extensive, its complicated reconstruction phase and high postoperative morbidity rates. From the patient's point of view, it is characterized by a principal change in the quality of life. In this case review the authors present their experience with a multidisciplinary approach to these procedures. PMID- 16218350 TI - [Analysis of surgical procedures on the vena saphena magna in the Czech Republic and an effect of Detralex during its stripping]. AB - The aim of this clinical study was to compare the degree of postoperative pain (VAS--10 cm, quality of life questionnaire CIVIQ and patient diary) between two groups of patients: patients treated with Detralex 14 days before and 14 days after the stripping of greater saphenous vein (GSV) and patients not treated with Detralex. In addition, the two groups were also compared for the incidence of symptoms associated with chronic venous insufficiency (CVI) (using the VAS scale: edema, tired and heavy legs, cramps, itching sensation), size of hematoma, use of analgesics and overall efficacy of the treatment. Clinical study included 181 patients from 15 medical centers throughout the Czech Republic. High ligation and partial stripping of greater saphenous vein on one lower extremity was performed in all patients (short stripping from groin to knee). Patients were randomly assigned in two groups: patients treated with Detralex (92) and patients not treated with Detralex (89). Patients in the first group were treated with Detralex for the period of 1 month. Degree of pain and patient's health condition were evaluated by the physician during D-14 (14 days prior to the surgery), D7 and D14 (7 and 14 days after the surgery) visits using the 10-cm visual analog scale VAS. The results indicate that Detralex reduced the intensity of postoperative pain, which resulted in decreased consumption of analgesics. Hematoma was smaller in patients already using Detralex 14 days prior to the scheduled stripping procedure. These patients also showed significant improvement of CVI symptoms and the quality of life of patients with CVI. High quality venoactive drugs administered 14 days prior to the surgery improve postoperative course in patients indicated for surgical treatment of varices. PMID- 16218351 TI - [Effect of laser irradiation of diode laser on healing of surgical wounds in rats]. AB - The aim of this work was to continue in previous study, which concerns biostimulation of skin wound healing evaluated after 24, 48, 120, 168 hours and so complete the chronological continuance of the process during the first seven days. Male, Sprague-Dawley rats (n=21) were used for the experiment. The rats were divided into 3 groups of 7 animals. In general anaesthesia (combination of xylazine, ketamine and tramadol) under aseptic condition two 3,5 cm long parallel skin incisions were performed on the left and right side of the rats spine and immediately sutured. The left wounds were daily stimulated with the diode laser (670 nm). The right wounds were not stimulated and served as control. The specimens of skin wounds were removed for histological evaluation 72, 96 and 144 hours after surgery. The biological specimens were stained with hematoxylin and eosin and histopathologically evaluated. In summary, in our histomorphological study of the influence of laser irradiation on primary wound healing evaluated after 72, 96 and 144 hours was concluded, that the healing of stimulated wounds was accelerated in comparison with controls. The histological evaluation showed earlier regress of inflammatory phase, faster finishing of reepithelization and acceleration in maturation phase. Presented experimental study completes the previous study and achieves the positive effect of biostimulation on all phases of skin wound healing in vivo. PMID- 16218352 TI - [An experimental model of the lymphatic vessels transplantation]. AB - The aim of this work is to describe a procedure of primary reconstruction of the lymphatic vessels on a model of heterotopic intestine transplantation in a rat. Both the donor and the recipient part of the procedure are described in a detail, as well as diagnostic methods of postoperative patency of the lympahatic anastomosis. In the end, the authors' own trial group is presented. However, no statistically significant differences were detected in histological findings of the intestinal graft rejection in rats with lymphatic anastomosis, compared with the control group. PMID- 16218353 TI - [Modification of surgical treatment of ingrowing toenail]. AB - The authors present their own modification of Emmert's operation used by treatment of ingrowing toenail. The modification itself consists in special technique of the suture of operation wound which prevents further nail incarnation. In second part the results of treatment of 51 patients are presented and discussed in context of other treatment modes. Patients treated by modified technique were treated approximately 14.06 +/- 5.44 days and needed approximately 3.79 +/- 1.04 checkups, others, treated by standard methods it was approximately 19.54 +/- 1.98 days and 4.73 +/- 0.62 checkups. The disposal of modified suture technique may reduce needed post-operation care. The question is if it is able to reduce the recurrence rate as well. PMID- 16218354 TI - [Contemporary surgical venous problematics for use by ambulatory surgeons]. PMID- 16218355 TI - [New concepts in treatment of gastric carcinoma]. AB - Despite interdisciplinary approach to gastric cancer the outcome of patients remains poor. Even after curative resection most patients suffer from progression of disease and metastasizing course. Adjuvant treatment like chemotherapy or radiation can not improve survival substantialy. This is discussed to be due to dissemination of tumor cells, which are not affected by most treatment options. The new concept of tumor specific antibody therapy with a monoclonal antibody is discussed as an effective treatment especially to fight disseminated tumor cells and thus might help to improve the outcome in stomach cancer. PMID- 16218357 TI - [Diurnal rhythms in internal medicine]. PMID- 16218358 TI - [Contemporary drug therapy: dramatic dangers and the optimism of ability]. PMID- 16218360 TI - [Topical aspects of arterial hypertension in clinical medicine]. PMID- 16218359 TI - [Milestones of the development of arterial pressure measurement methods (on the occasion of the 100th anniversary of N S Korotkov sound method of arterial pressure measurement)]. PMID- 16218361 TI - [Arterial hypertension and body weight: the solved and unsolved problems]. PMID- 16218362 TI - [Diabetes mellitus and the heart]. PMID- 16218363 TI - [Genetic risk factors of thyroid gland pathology]. PMID- 16218364 TI - [Significance of antibodies to cyclic citrullinized peptide in diagnostics of rheumatoid arthritis]. PMID- 16218365 TI - [Correction of thrombocyte hemostasis and biological age reduction in metabolic syndrome]. AB - The study shows that application of metformin, hypocaloric diet, and rational physical loads in patients with arterial hypertension (AH) with metabolic syndrome and impaired glucose tolerance (IGT), reduces pathologically hightened biological age (BA), eliminating shifts in blood lipid spectrum, moderating peroxidation syndrome, and reducing changes in thrombocyte homeostasis. Such complex therapy normalizes the adhesive and aggregation function of thrombocytes in vitro, and their intravascular activity. The parameters of the subjects had approximated the control level by the end of the 24th week of treatment. To reduce the biological age down to the level of chronological one, stabilize metabolic processes and thrombocyte homeostasis in patients with AH and IGT, the offered therapy should be prolonged and controlled. PMID- 16218366 TI - [On the sodium to potassium ratio in daily urine of patients with arterial hypertension]. AB - The aim of the study was to measure Na:K ratio in 24-hour urine depending on salt consumption. The subjects were 230 patients with arterial hypertension (AH). The study found a significantly higher salt consumption by AH patients, and decrease of K excretion together with increase of Na excretion. Measurement of Na:K ratio in these patients demonstrated that their salt consumption should be limited, and the proportion of food containing K should be increased. PMID- 16218367 TI - [Risk of arterial hypertension and social support]. AB - The aim of the study was to investigate the effect of social support on the 8 year risk of arterial hypertension (AH) in men aged 25 to 64. The researchers studied a random representative sample of male residents of a Novosibirsk district aged 25 to 64 years. The level of social support was evaluated by Berkman-Syme method. All the cases of AH which occurred in the cohort within 8 years (1994-2002) were investigated. The statistical analysis was performed using SPSS-11.5 software package. The relative risk (RR) of AH was evaluated using a Cox proportional regressive model. Within these 8 years the RR of AH was higher: 1) in men aged 25 to 64 years with low and medium close contact index (CCI) compared to men with high CCI (8.5 times); 2) in subjects with low social contact index (SCI) compared to subjects with high or medium SCI (4.8 times). In men aged 55 to 64 years the RR of AH was 15.9 times higher in men with low CCI. The highest rate of AH was found in men with low CCI and SCI belonging to the groups of widowers, divorced people, subjects with incomplete secondary or primary education, as well as in pensioners and people with low level of social support. The results show that AH risk in men aged 25 to 64 years is associated with low level of social support. PMID- 16218368 TI - [NO synthesis in the vascular endothelium of patients with type II diabetes]. AB - The subjects of the study were 30 patients with type II diabetes and arterial hypertension. The purpose of the study was comparative clinical characterization of type II diabetes patients with and without microalbuminuria (MAU), and evaluation of effects of 16-week therapy aimed at stimulation of NO synthesis by vascular endothelium, on the pathogenesis of diabetic nephropathy. The results show that patients with MAU, unlike those without it, are characterized by longer course of diabetes, more pronounced lipid exchange disorder, more variable arterial pressure, higher pressure load index, elevated activity of lipid peroxidation (LP) processes, prominent disorder of NO-producing endothelial function. After 16 weeks of treatment, both groups demonstrated significant increase of NO basal secretion, prominent reduction of arterial hypertension, significant improvement of important carbohydrate and lipid exchange parameters, reduction of LP activity, and thus reduction of MAU. The listed changes were more significant in patients with MAU. PMID- 16218369 TI - [Myocardial remodeling in chronic renal failure patients on hemodialysis]. AB - Pathologic myocardial changes in is one of the main causes of death in patients with terminal chronic renal failure (CRF). The authors studied structural and functional condition of the myocardium and the effect of various CRF factors on cardiovascular complications. The study demonstrates positive correlation between systolic arterial pressure (AP) and left ventricular (LV) remodeling, as well as between the degree of anemia and LV remodeling in patients with CRF on program hemodialysis. Concentric LV hypertrophy (LVH) was revealed in 24 (65%) patients, eccentric LVH--in 7 (19%) patients. The results show that eccentric LVH with reduced ejection fraction in CRF patients on hemodialysis is associated with high systolic and pulse pressure. PMID- 16218370 TI - [Efficacy of galavit in patients with duodenal ulcer]. AB - The aim of the study was to investigate clinical efficacy of the medical complex Galavit in patients with acute phase of duodenal ulcer (DU) in. The subjects were 60 DU patients aged 32 +/- 2 years with ulcerous defects of 0.4 to 1.3 cm in diameter. In patients receiving Galavit, pain was coped with by Mann-Whitney method in 2.5 +/- 0.2 days, p < 0.001; in the control group--in 5.7 +/- 0.1 days. In the Galavit group the ulcers healed in 11.3 +/- 0.2 days, p < 0.001; in the control group--17.8 +/- 0.3 days; in 4 cases (13.3%) the ulcerous defects healed with forming of rough scars. Galavit elevated T-lymphocyte rate from 53.1 +/- 0.6% to 65.1 +/- 0.2%, p < 0.001; T-helper inductor level--from 27.8 +/- 0.2% to 38.5 +/- 0.3%, p < 0.001; cytotoxic T-lymphocyte level--from 18.5 +/- 0.5% to 27.3 +/- 0.3%, p < 0.001; B-lymphocyte level--from 12.3 +/- 0.2% to 19.1 +/- 0.1%, p < 0.001. The therapy significantly lowered malonic aldehyde level by 23.5%, trienoic conjugate level--by 61.6%; superoxide dismutase level rose 1.6 times, catalase level--1.4 times, glutathion reductase level--from 19.03 +/- 1.17 to 27.01 +/- 1.24 optical density units/mg, p < 0.001. The study did not find any significant changes in the immune status and lipid peroxidative/antioxidative system of patients receiving basic therapy. The results show that Galavit has anti-inflammatory effect, improves immune status and anti-oxidative protection. It is appropriate to administer Galavit as a component of DU basic therapy. The results show that Galavit has anti-inflammatory effect, improves immune status and anti-oxidative protection. It is appropriate to administer Galavit as a component of DU basic therapy. PMID- 16218371 TI - [Alimentary tract involvement in Behcet's disease]. PMID- 16218372 TI - [Irritable bowel syndrome: moot points and unanswered questions]. PMID- 16218373 TI - [The five paradoxes of the contemporary stroke prevention conception]. PMID- 16218374 TI - [Hypertensic crises]. PMID- 16218375 TI - [On the history of study and teaching of infectious diseases in Moscow State University (on the occasion of the 250th anniversary of M V Lomonosov Moscow State University)]. PMID- 16218376 TI - [Gene expression profiling of migraine]. PMID- 16218377 TI - [An overview of the revised international classification of headache disorders (ICHD -II)]. AB - The International Headache Society (IHS) revised The International Classification of Headache published in 1988. Old version has facilitated epidemiological and multinational clinical trials. Fifteen years after its original publication, a revised International Classification of Headache Disorders 2nd Edition(ICHD-II) has been unveiled in 2004. Modifications are small but significant. A lot of knowledge and evidences were added to the new version. A revised classification classifies headaches in 14 groups and categorized headache groups into four parts; part 1: primary headache, part 2: secondary headache, part 3: neuralgias and facial pain, and appendix. Physicians should continue to consult the IHS criteria to ensure accurate diagnosis and management for headache. PMID- 16218378 TI - [Historical background of researches in headache]. AB - In seventeenth century, Thomas Willis pointed out the source of headache is distention of the vessels nearly identical with Wolf's twentieth century vascular hypothesis of migraine. The concept of the pathophysiology in migraine swung to neural cause by Edward Liveing (1873). The research evidence for special role of serotonin was revealed by modern approach and the mechanism of migraine was presented in trigeminovascular system. The development of the basic science leads to new treatments in headache clinic. PMID- 16218379 TI - [Epidemiology of headache]. AB - We discuss here the epidemiology of chronic headache. Headache is a widespread and costly public health problem. Few people do not experience headache: in men, the lifetime prevalence for headache of any kind is 93 %, and for women it is up to 99%. Approximately 8.4 million people in Japan suffer from migraine, and 22 million have tension-type headache. Despite the painful, costly, and disabling impact of headache, many patients with headache do not seek medical advice. It is important to recognize the incidence of various kinds of chronic headache, and to diagnose and treat them correctly. In this article, we review the incidence, precipitating factors, regional prevalence, and age dependence of the incidence of each type of chronic headache. PMID- 16218380 TI - [Structure and function of nociceptive neuronal receptors]. AB - It has been well known that pain is caused by nociceptive stimulation such as protons (i.e. acidic solutions), heat and capsaicin, a pungent ingredient of chilli peppers. For a long period, the signal transduction mechanism of pain activated by these nociceptive stimuli has not been clarified. Recent advance, especially the identification of TRPV1 receptor (for which capsaicin, protons and heat are ligands), P2X and P2Y receptor (for which ATP is a ligand) and acid sensing ion channel made a remarkable progress in understanding the mechanism of nociceptive neurons. This article reviews the structures and functions of nociceptive neuronal system particularly in TRPV1 receptor, P2X and P2Y receptors. PMID- 16218381 TI - [Migraine headache and mitochondrial DNA abnormality]. AB - Migraine headache is a heterogenous group of neurologic disorders with high prevalence in the general population and strong familial aggregation. Epidemiologic evidence for frequent maternal transmission has long suggested a role for an altered mitochondrial genetic background. This is supported by the observation of impaired energy metabolism and mitochondrial function in migraine patients as well as by the frequent association of migraine headache in MELAS. In the literature, some patients having migraine headache are reported to have mitochondrial DNA abnormalities, however, systematic screening studies have failed to demonstrate the clear relationship between migraine headache and mitochondrial DNA abnormalities. Migraine may be associated with mutations of nuclear genes which encode respiratory chain enzymes. PMID- 16218382 TI - [Genetic analysis of migraine headache: a review]. AB - Recent advances in genetic analysis of migraine headache are reviewed. Point mutations of P/Q -type Ca2+ channel alpha1 subunit(CACNA1A) gene and Na-K ATPase, alpha2 (ATP1A2) gene have been identified in the familial hemiplegic migraine (FHM-1 and FHM-2, respectively). Mutations in notch-3 gene cause the cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), which is an autosomal dominant inherited disorder often accompanying with migraine like headache. Serotonin (5-HT) related genes, dopamine D2 receptors (DRD2) gene, methylenetetrahydrofolate reductase (MTHFR) gene, and angiotensin converting enzyme (ACE) gene have been noticed as the susceptible genes for migraine pathogenesis. Genetic study of migraine is promising and will provide further understanding of the migraine pathophysiology. Discovery of the responsible or susceptible genes will open an avenue to develop new therapeutic strategy. PMID- 16218383 TI - [Migraine update]. AB - Migraine is one of the common diseases suffering 8.4 million patients in Japan. The pathophysiology of migraine remains unclear. The genetic and basic studies of the familial hemiplegic migraine, a specific subtype of migraine with aura, have demonstrated the dysfunction of mutant brain-expressed calcium ion channel and/or the Na+/K+ ion transporter and suggested the association between cortical spreading depression (CSD) and migraine with aura. It is suggested that the CSD, neurogenic inflammation and vasodilatation caused by unknown triggers may activate the 'brainstem migraine generator' and amplified back way. In consequence, headache and/or aura will be appeared and strengthened. Our etiological data of headache in Daisen located in Western Japan clarified as follows; 1) Overall prevalence of migraine in Daisen was 6.0%. Women observed a 5.9-fold higher risk of migraine than men. 2) Fatigue, mental stress, and lack of sleep were the main headache triggers. 3) Only 7.3% of those with migraine with aura and 5.3% of those with migraine without aura had consulted a physician. 4) Migraineurs consume significantly more fatty/oily foods, coffee, and tea than nonheadache subjects of the same community. Migraineurs consume significantly fewer fish than nonheadache residents. As a conclusion, only a few Japanese migraineurs receive benefits of medical services and recent advances of headache medicine. The Japanese guideline for chronic headache treatment has declared in 2002. The International Classification of headache disorders has reedited to the 2nd edition. Public education concerning headaches is one of the most urgent issues in Japan. PMID- 16218385 TI - [Cluster headache]. AB - Cluster headaches are characterized by attacks of strictly unilateral severe pain which is orbital, supraorbital, temporal or in any combination of these sites associated with one or more of the following, ipsilateral conjunctival injection, lacrimation, nasal congestion, rhinorrhea, forehead and facial sweating, miosis, ptosis, and eyelid edema. Most patients are usually unable to lie down, restless or agitated during an attack. The attacks last 15-180 minutes and occur from once every other day to 8 times a day. Cluster periods usually last for weeks or months separated by remission periods, however about 10-15% of patients have chronic symptoms without remission. Age at onset is usually 20-40 years and prevalence is 3-4 times higher in men than in women. Acute attacks involve activation of the posterior hypothalamic gray matter. Pharmacological treatment for cluster headache can be abortive, prophylactic, or a combination of both methods. PMID- 16218384 TI - [Tension-type headache]. AB - Tension-type headache is the most common type of primary headache. Life time prevalence of tension-type headache in general population ranges from 30 to 78%. It is the least studied of the primary headache disorders, despite the fact that it has the highest socio-economic impact. In the new 'The International Classification of Headache Disorders', tension-type headache is divided into episodic and chronic subtypes. The exact mechanisms of tension -type headache are not known. Peripheral pain mechanisms are most likely to play a role in episodic headache, whereas central pain mechanisms play a more important role in chronic tension-type headache. Treatments of tension-type headache include not only medication but also the physical therapy. In many uncertain cases there is overuse of medication. Two months after medication overuse has ceased, chronic tension-type should be diagnosed. PMID- 16218386 TI - [Other primary headaches]. AB - Most primary headaches are classified into a few categories, such as migraine or muscle contraction headache, and patients suffering from these headaches are common. On the other hand, other primary headaches are very rare. In this section entitled "Other primary headaches", eight headaches, including primary stabbing headache, primary cough headache, primary exertional headache, primary headache associated with sexual activity, hypnic headache, primary thunderclap headache, hemicrania continua, and new daily-persistent headache, are described. Some characteristics of other primary headaches are common in symptomatic headaches, such as subarachnoid hemorrhage or arterial dissection. Therefore, careful evaluations including neuroimaging are necessary to exclude organic diseases. PMID- 16218387 TI - [Headache attributed neurosurgical diseases]. AB - When we see headache patients, it is most important thing that we rule out patients with intracranial diseases. Intracranial diseases with headache are so many. I'd like to emphasize some intracranial diseases with headache not to miss. Especially subarachnoid hemorrhage due to ruptured cerebral aneurysm is a serious disease not to miss. Recently the cases of dissecting cerebral aneurysm (vertebral artery) and venous/sinus thrombosis are increasing with spreading MRI/MRA examination. And brain tumors and chronic subdural hematoma is a serious disease not to miss in recent Japanese aging society. I'd like to mention these important diseases with headache. PMID- 16218388 TI - [Secondary headache in children]. AB - Headache is one of the common symptoms of fever and headache without fever is also not rare in general pediatric clinics. The common causes of these headaches involve extracranial infection due to viral illness, migraine and trauma. Headache with vomiting, fever and meningeal signs suggests meningitis. Taking blood pressure is necessary, even though headache caused by hypertension is rare in children. Neuroimaging should be performed with abnormal neurological findings, atypical headache pattern, or significant change of preexisting headache. Serious underlying diseases, such as brain tumor or intracranial hemorrhage, are uncommon, however they should be diagnosed immediately using neuroimaging because of their urgency. PMID- 16218389 TI - [The characteristics of sinus headache resembling the primary headaches]. AB - It has to be excluded organic lesions to diagnose the primary headache, however they are tended to be misdiagnosed in a routine practice. Acute sinusitis is the most common disease to be misdiagnosed as the primary headaches and we have reported that the characteristics of sinus headache have closely resembled migraine, cluster headache or tension type headache. The effectiveness of triptans does not become an evidence for a diagnosis of migraine or cluster headache, because it was also effective for the pain of the acute sinusitis. The etiology of sinus headache that resembles the primary headaches is similar to the trigemino-vascular theory. In this paper, we clarify the characteristics of sinus headache resembling the primary headaches. PMID- 16218390 TI - [The secondary headache in dentistry]. AB - This report summarizes the secondary headache attributed to dental disease. Among a lot of dental diseases, the temporomandibular disorder(TMD) is the most important for understanding the mechanism of this type of headache. Our recent study showed that TMD patients had the following symptoms: dysdiadochokinesia, abnormal induced-rigidity in forearms, abnormal circadian rhythm, tension-type headache and mental torment. All these abnormal symptoms were relieved by the improvement of the patient's sleep-quality. The brain monoaminergic systems are known to be related to muscle tonus, sleep problems and mental symptoms. Our studies of a serotonin transporter(5-HTT) gene promoter polymorphism showed that the L and XL alleles were more frequent and S allele was less frequent in the TMD compared to the controls. We speculate the brain monoaminergic systems play the important pathophysiological roles on the TMD. PMID- 16218391 TI - [Headache in female]. AB - Significant correlation is observed between the prevalence of headache (tension type headache and migraine) and such variables as gender and age. The prevalence of migraine in reproductive-aged females is 4 to 5 times higher than that in males. However, the prevalence in headache does not differ with gender before puberty. Female life stages include the pre-pubertal period, pubertal period, reproductive period (pregnancy, parturition, and puerperium), menopausal period, and postmenopausal period. During these life stages, serum estrogen levels change dynamically. Even during the menstrual cycle, serum estradiol levels change dynamically. These changes definitely differ from changes in serum testosterone levels in males. Headache or migraine during the menstrual cycle is more frequently observed during the period of menstruation. However, the prevalence of headache and migraine become lower after menopause. These findings suggest that the occurrence of headache is in part associated with the decline in serum estradiol levels. PMID- 16218392 TI - [Ophthalmology]. AB - The Headache Classification Subcommittee of the International Headache Society classifies headaches related to eyes as "Headache attributed to disorder of eyes" in the International Classification of Headache Disorders; 2nd Edition(ICHD-II). It consists of "Headache attributed to acute glaucoma", "Headache attributed to refractive errors", "Headache attributed to heterophoria or heterotropia(latent or manifest squint)", "Headache attributed to ocular inflammatory disorder". But other causes of headache related to eyes exist. For example, dry eye causes the headache. This article mentions to "Headache attributed to disorder of eyes" in ICHD-II, and additionally, describes other causes of headache associated with disease of eye. PMID- 16218393 TI - [The role of triptans and analgesics for primary headache treatment]. AB - Primary headache especially migraine is very common disorder. The mainstay in the acute treatment of migraine is triptans (sumatriptan, zolmitriptan, eletriptan, naratriptan) and analgesics or NSAIDs. However, it is still unclear the appropriate usage of triptans and analgesics or NSAIDs for migraine treatment. Mild attacks may be managed with analgesics or NSAIDs while severe disabling ones usually respond better to specific antimigraine drugs, triptans. Analgesics or NSAIDs administration is always plagued with the potential of subsequent drug induced headache phenomenon. Therefore usage of analgesics or NSAIDs should be restricted only for young and typical type patients with migraine. As triptan medication method corresponding to various life style, in addition to tablet formulation, there are subcutaneous injector and nasal spray formulation in sumatriptan, rapid melt tablet formulation in zolmitriptan (rapimelt) and naratriptan (rapidisk). These different type of formulation are valuable for patient's needs. PMID- 16218394 TI - [Management and treatment of headache in the pain clinic]. AB - Nerve block therapy is one of the non-pharmacological methods in headache treatment. Most pain clinics in Japan use nerve block therapy to treat pain together with pharmacological methods. Sensory nerve blocks such as the trigger point block, occipital nerve block, trigeminal nerve block, C-2, 3 spinal nerve block, etc., are effective for headache pain. One of the characteristic treatments of headache in the pain clinic in Japan is the stellate ganglion block(SGB). Although it has not been fully clarified why SGB is effective in treating headache, stabilization of the abnormal sympathetic nerve function, suppression of inflammation of vascular wall etc., are thought to be related to the effectiveness of SGB. Nerve block therapy is effective for the treatment of headache patients and offers us an important therapeutic option for treatment of headache pain. PMID- 16218395 TI - [Prophylactic treatment of migraine]. AB - The aim of prophylactic treatment of migraine is to reduce the frequency and severity of migraine attacks. Identifying relevant trigger factors can help reduce the frequency of migraine attacks. The headache diary is useful to identify trigger factors and pattern of headaches and to assess the efficacy of medication. Prophylactic drugs should be considered when attacks are frequent or severe and the acute treatments such as triptans or NSAIDs are not effective. Lomerizine, propranolol, valproate and amitriptyline are useful. Lomerizine is recommended as the first-line prophylactic drug because it is licensed as a preventive drug for migraine in Japan. If attacks has not improved after using a prophylactic drug for 2 months, the drug can be changed to another drug. Propranolol is particularly useful if a patient has hypertension. Amitriptyline is useful if there is associated depression and/or tension-type headache. Valproate is considered if attacks are frequent. Patients should be informed benefits and potential side-effects of the medicine. PMID- 16218396 TI - [Prophylactic treatment with botulinum toxin type A]. AB - Migraine is a common neurological disorder, characterized by recurrent attacks of severe headache. Its prevalence is estimated 8.4% of general population in Japan. Acute migraine treatment has dramatically improved with the development of a novel class of selective 5-HT1B/1D/1F receptor agonists, known as the triptans, but prophylactic treatment still remains limited. Botulinum toxin type A(BoNT-A) has recently been shown to be effective in randomized control trials, but no prospective study has been conducted yet in Japan. In this paper, we review the results of recently published controlled trials, touching on its methodology, optimal dose, patient selection, and possible mechanisms, as well as on going clinical trial in our university. PMID- 16218397 TI - [Medication-overuse headache]. AB - Medication-overuse headache(MOH) is a clinically important entity and it is well documented that the regular use of acute symptomatic medication by patients with migraine or tension type headache increases the risk of aggravation of the primary headache disorders. MOH is one of the most common causes of chronic refractory headache. The pathophysiological mechanism of MOH is still unclear. But as in most of the headache entities, several different aspects, such as genetic background, peripheral and central nervous system interaction, specific psychotropic effects, appear to play key roles. Management of MOH is a difficult problem. The education for patients with MOH is very important. PMID- 16218398 TI - [Clinical condition and treatment of combined headache]. AB - Coexistence of migraine with tension-type headache, that is, so-called combined headache is a disabling chronic headache disorder and is often seen in daily practice. Combined headache is a common disease, but it is not so easy to do an accurate diagnosis and adequate treatment because clinical spectrum is wide. For clinician, it is especially important to understand characteristics of patient's headache enough to diagnose correctly by using such as a headache diary and, to educate patients to differentiate between migraine and tension-type headache in order to select the right treatment and to prevent medication-overuse headache. PMID- 16218399 TI - [Diagnostic imaging of headache]. AB - The diagnosis of headache is based on the subjective symptom, and it is generally classified into the primary headaches, which have intracranially no abnormality, and the secondary headaches, which have organic abnormality. Neuroimaging examinations, computed tomography (CT) scan, and magnetic resonance imaging (MRI) are useful for differential diagnosis and can perform to rule out vascular malformation, brain tumor, and other pathologies, and to confirm the diagnosis. There are cases of the secondary headaches resembling the primary headaches and patients complaint of pain by complex of them, in those cases, it is necessary to perform more detailed neuroimaging(three dimensional MRA, helical CT and so on). These neuroimaging could contribute to make diagnosis and treatments for the patients. PMID- 16218400 TI - [Relationship between cyclooxygenase (COX)-2 and malignant tumors]. AB - Recently, epidemiologic studies and animal experiments have demonstrated that nonsteroidal anti-inflammatory drugs(NSAIDs) reduce the incidence of colorectal carcinoma. Cyclooxygenase(COX) is the principal target of NSAIDs. COX is the first oxidase in the process of prostaglandins(PGs) production from arachidonic acid(AA). PGs and COX enzyme may be involved in the initiation and/or the promotion of carcinogenesis because the major action of NSAIDs is the inhibition of COX. In this review, we demonstrated the expression of COX-2 in urological cancer(renal cell carcinoma, bladder tumor, prostate cancer, and testicular tumor) tissues as well as the effects of COX inhibitors. PMID- 16218401 TI - [Postprandial hyperglycemia as a risk factor of atherosclerosis]. PMID- 16218402 TI - [Efficacy and problems of bladder volume measurement using portable three dimensional ultrasound scanning device--in particular, on measuring bladder volume lower than 100ml]. AB - OBJECTIVES: Using a portable three dimensional ultrasound scanning device (The Bladder Scan BVI6100, Diagnostic Ultrasound Corporation), we examined measured values of bladder volume, especially focusing on volume lower than 100 ml. MATERIALS AND METHODS: A total of 100 patients (male: 66, female: 34) were enrolled in the study. We made a comparison study between the measured value (the average of three measurements of bladder urine volume after a trial in male and female modes) using BVI6100, and the actual measured value of the sample obtained by urethral catheterization in each patient. We examined the factors which could increase the error rate. We also introduced the effective techniques to reduce measurement errors. RESULTS: The actual measured values in all patients correlated well with the average value of three measurements after a trial in a male mode of the BVI6100. The correlation coefficient was 0.887, the error rate was--4.6 +/- 24.5%, and the average coefficient of variation was 15.2. It was observed that the measurement result using the BVI6100 is influenced by patient side factors (extracted edges between bladder wall and urine, thickened bladder wall, irregular bladder wall, flattened rate of bladder, mistaking prostate for bladder in male, mistaking bladder for uterus in a female mode, etc.) or examiner side factors (angle between BVI and abdominal wall, compatibility between abdominal wall and ultrasound probe, controlling deflection while using probe, etc). CONCLUSIONS: When appropriate patients are chosen and proper measurement is performed, BVI6100 provides significantly higher accuracy in determining bladder volume, compared with existing abdominal ultrasound methods. BVI6100 is a convenient and extremely effective device also for the measurement of bladder urine over 100 ml. PMID- 16218403 TI - [Measurement of salivary testosterone by a simple enzyme immunoassay procedure]. AB - PURPOSE: The level of serum free testosterone (Se free T) is thought to affect personal emotion and behavior. On the other hand, salivary testosterone (Sa-T) exists mainly in an unbound state and its measurement is noninvasive and repeatable. The aim of this study was to evaluate whether Sa-T can be measured in a clinical setting using a simple enzyme immunoassay kit, and to evaluate whether the level of Sa-T obtained by a simple procedure can be used as a substitute for the level of Se free T by comparing the values obtained with those of Se free T. PATIENTS AND METHODS: Seventy-six volunteers from among patients who visited our outpatient clinic. They were 74 male and 2 female patients between the ages of 28 and 90 (mean 67.2 +/- 12.7 years, median 70 years). Sixteen of 74 male patients were treated with hormone therapy for prostate cancer. All of 76 patients were scheduled for blood examinations because of urological diseases, and gave full informed consent to participate in this study. Saliva and serum samples were collected between 06.00 and 11.00 h. Sa-T, Se free T and the concentration of transferine in saliva was measured with a Salivary Testosterone Enzyme Immunoassay Kit, a Coat-A-Count Free Testosterone and a Blood Contamination Enzyme Immunoassay Kit, respectively. The average of duplicate values was used. Bioavailable testosterone (BT) was assayed by Liquid Chromatography-Mass Spectrometry/Mass Spectrometry. Average intraassay coefficient of variations of all assays were within 15%. RESULTS: Eleven out of 76 salivary samples were disqualified as unsuitable because of possible blood contamination or falling outside the range of the assay sensitivity. The ranges of Sa-T, Se free T and the transferrin concentration in saliva were 32 to 360 pg/ml (n = 65, mean 176.8 +/- 96.6 pg/ml, median 168.0 pg/ml), 0.15 to 21.0 pg/ml (n = 76, mean 6.6 +/- 4.2 pg/ml, median 7.0 pg/ml), and 0.08 to 6.0 mg/dl (n = 76, mean 0.66 +/- 0.89 pg/ml median 0.42 mg/dl), respectively. In 10 participants randomized chosen out of all, there was significantly correlation between Se free T and BT (r = 0.964, p < 0.01) . There was significant correlation between Se free T and Sa-T (n = 65, r = 0.592, p < 0.01) and estimated Se free T value can be calculated based on the Sa T level. CONCLUSION: Though disqualified samples accounted for 14.5% of the total number of saliva samples, Sa-T can be used as a substitute parameter for biologically active testosterone. PMID- 16218404 TI - [Uncertain analgetic efficacy of intraurethral instillation of anesthetic jelly in rigid cystoscopy for men]. AB - PURPOSE: The analgetic efficacy of intraurethral instillation of anesthetic jelly in rigid cystoscopic examination for men is not clearly confirmed. This study evaluated the usefulness of intraurethral instillation of anesthetic jelly by questionnaires regarding the pain and uncomfortable feelings during and after the cystoscopic examination. MATERIALS AND METHODS: A total of 62 men were enrolled in the study. Sixteen patients had not received cystoscopy previously (first cases), and 46 had prior experiences of cystoscopy more than twice (repeat cases). These patients were sequentially divided into two groups; one group (jelly-group) received intraurethral instillation of oxybuprocaine hydrochroride jelly (Benoxil jelly) prior to cystoscopic examination, and the other group (no jelly-group) did not receive the jelly instillation. Pain during the jelly instillation to examination, and uncomfortable feelings at micturition after the examination was evaluated by questionnaires. RESULTS: Instillation of the jelly itself caused considerable pain, which almost similar to that of scope insertion. There was no significant difference in pain score between jelly-group and no jelly-group during the insertion of the scope to examination. The miction pain after examination was significantly severer in the jelly-group than in the no jelly-group in repeat cases. In addition, 75% of repeat cases felt better comfortableness when jelly was not instilled, and 96% of repeat cases did not want jelly instillation for their future examination. CONCLUSIONS: Intraurethral instillation of anesthetic jelly had no efficacy for reducing the pain during cystoscopy. Instillation of the jelly itself caused considerable pain, and it made pain and uncomfortable feelings after the examination worse. These results suggested that the instillation of anesthetic jelly before rigid cystoscopy in men might not be useful enough for reducing the uncomfortable feeling during the examination. PMID- 16218405 TI - [Roles of BPH impact index in the evaluation of impaired urination in patients with BPH]. AB - OBJECTIVE: To assess the significance and current status of the benign prostatic hyperplasia (BPH) impact index (BII) in the evaluation of subjective symptoms of impaired urination in so-called QOL disease, BPH. PATIENTS AND METHODS: Over the past 2 year-period, in 159 patients with the diagnosis of BPH were asked to reply to each of the international prostate symptom score (I-PSS), QOL index and BII questionnaires. The subjective symptom scores (a total of 246 points) were evaluated from the viewpoint of clinical statistics in the search for any these and to find which questions cover the BII, most. RESULTS: 1) Statistically significant but moderate correlations were observed among I-PSS total score, QOL index and BII. The correlations among Qmax, BII and QOL were very weak. 2) Out of the 11 domains in both IPSS and BII, 2 questions of BII ("bothersomeness caused by urinary problems" and "degree of worry about well-being") and 4 questions of IPSS ("residual sense," " pollakisuria," "weak urinary stream" and "nocturia") were shared as QOL indices. Patient satisfaction was affected also by the questions in the BII. 3) Of the 7 BPH symptoms assessed in IPSS, 4 symptoms ("residual sense," "pollakisuria," "weak urinary stream" and "nocturia") affected the QOL index, and 4 symptoms ("urgency on micturition," "residual sense," "nocturia" and "strain at urination") affected BII. 4) Of the 7 symptoms assessed by IPSS, different symptoms affected each of the 4 BII questions. CONCLUSION: It is needed to assess BPH symptoms not only by the IPSS and QOL index but also based on BII to provide the detailed therapeutic instructions and thorough patients consultation. PMID- 16218406 TI - [Descriptions of urinary stone in the Hippocratic collection]. AB - PURPOSE: Hippocrates was a famous physician of Kos in ancient Greece (c. 460 B.C.). His works later were described in the Hippocratic Collection, in which I studied references to urinary stones. MATERIALS AND METHODS: I collected mentions of urinary stones from the Hippocratic Collection (Roeb, Otsuki, and Kon editions) to compared descriptions of urinary stones between the Kos and Knidos schools. RESULTS: Urinary stones were mentioned in 24 passages of the collection: 12 (50%) referred to pathogenesis of urinary stones; 6 (25%) to symptoms; 4 (17%) to treatment, and 2 (8%) to other aspects. Symptoms of urinary stones included hematuria, colic, painful urination, difficult urination, and passing a urinary stone. Bladder stones were mentioned in 15 passages (63%), renal stones in 4 (17%), and both bladder and renal stones in 2 (8%). Although no site was mentioned directly in 7 passages (29%), these appeared most likely to refer to bladder stones. Descriptions by Kos, Knidos, and unspecified schools accounted for 15 (63%), 4 (17%), and 5 (21%) of mentions of urinary stones. Descriptions of bladder stones by the Kos school were relatively numerous, while most of the few renal stones were described by the Knidos school. Treatments for urinary stone included medication, presumably to aid in passing the stone or to relieve pain; warm soaks or applications to treat pain; and incision over the kidney, when the affected area showed swelling with elevation. CONCLUSIONS: Urinary stones were mentioned in the Hippocratic Collection more frequently by the Kos school. No description of cystolitotomy included. The Oath of Hippocrates proscribed use of the knife to treat urinary stones, however, incision over the kidney (presumably nephrolithotomy) was mentioned in "Internal Affection" by the Knidos school. PMID- 16218407 TI - [A case report of sarcomatoid carcinoma of the bladder with metastasis to small intestine]. AB - A 67-year-old male presented to our clinic with gross hematuria. Cystoscopic examination revealed a broad-based tumor of 2.5 cm in diameter on the lateral side of the right ureteral orifice. Under the clinical diagnosis of TCC G2 > G3, T3bNOM0, radical cystectomy with orthotopic bladder substitution was performed. Pathological diagnosis was TCC G3 with sarcomatoid carcinoma, pT2pR0pL1 pVlpN0. Adjuvant chemotherapy was not performed because of his transient poor conditions. Lung metastasis was observed 6 months postoperatively. Despite of M-VAC therapy and radiation therapy, additional metastases to brain and liver were observed. One month later, partial ileectomy specimen for occlusive ileum revealed the same histologic findings, TCC G3 with sarcomatoid carcinoma. He died 9 months postoperatively. To our knowledge, this is the first case of sarcomatoid carcinoma of the bladder with metastasis to small intestine, although 6 cases of transitional cell carcinoma of the bladder with metastasis to small intestine has been reported in Japan. PMID- 16218408 TI - [A case of malignant lymphoma of the prostate]. AB - We report a case of primary malignant lymphoma of the prostate. A 76-year-old man was refer to our clinic with a chief complaint of dysuria. Based on a benign prostatic hypertrophy, transurethral resection of the prostate was performed. Histological findings showed diffuse, large cell type malignant lymphoma. His clinical stage was 1 A prostate (+) according to the Ann Abor classification. The combination chemotherapy with THP-COP was performed for 3 courses, followed by irradiation at 30 Gy. His prostate has showed no recurrence for 10 month after treatment. Primary malignant lymphoma of the prostate is rare. About 30 cases have been reported in the japanese literature. Preoperative diagnosis of malignant lymphoma of the prostate is difficult. All cases have been diagnosed after prostatic surgery or biopsy. PMID- 16218409 TI - [Bilateral adrenal lymphoma with adrenal insufficiency: a case report]. AB - A 65 year-old man visited the outpatient clinic of internal medicine complaining of fever and upper abdominal pain. Bilateral adrenal tumors were detected by CT scan and ultrasonography without any hormonal abnormalities. The diagnosis was bilateral primary non-functioning adrenal tumors because there was no tumor except adrenals with the detailed examinations. Then we are informed about the patient and he was transferred for the surgery. Cortisol was administrated because adrenal insufficiency was observed before surgery. Right adrenalectomy was performed after the recovery of patient status with cortisol. The pathological diagnosis was malignant lymphoma (diffuse large, B cell type). The patient was getting worse and dead 15 days after the surgery. PMID- 16218410 TI - [A case of sarcomatoid carcinoma in the region of the female urethra]. AB - A 78-year-old woman was presented with bleeding in the region of the genitalia. The tumor arose from the urinary meatus and biopsy was performed. The pathological examination revealed sarcomatoid carcinoma which was composed of squamous cell carcinoma and spindle cell sarcoma. She underwent urethrectomy and suprapubic cystostomy. She is alive at 26-month follow-up, after local radiation therapy of 50 Gy to the recurrence sites. PMID- 16218411 TI - [Relationship between physical restraint in nursing homes and their institutional characteristics]. AB - OBJECTS: To clarify relationship between physical restrain in nursing home and its institutional characteristics. METHODS: The subjects consisted of all 103 nursing homes in Okayama prefecture which were open in 2002 with a total of 6,829 residents. Data were obtained by questionnaire through the "Welfare ombudsman Okayama" in 2002 and audited by local government in the same year. We then performed multiple regression analysis. The parameter in question was the ratio of residents who were physically restrained to the residents at high risk of needing physically restraint (physically restrained ratio). Independent valuables were institutional characteristics. To increase the accuracy of estimation, we hypothesized that the residents likely to be the physically restrained were elderly with serious dementia who were defined as IV and over in the dementia elderly daily living independence rank. We also performed analysis with the definition of heavy dementia elderly graded as III and over. RESULTS: We obtained data of 72 institutes housing 1,700 elderly with serious dementia, which did not have any missing values. Of these 74% had at least one resident being physically restrained. The average physically restrained ratio was 24.2%. The institutional characteristics which correlated with use of physical restraint were intensive staffing, holding periodical care conference, and having unit care. The results were robust with regard to change in the serious dementia definition. CONCLUSION: The institutional characteristics which correlate with use of physical restraint are intensive staffing, holding periodical care conference, and having unit care. PMID- 16218412 TI - [Indoor air pollution in newly built or renovated elementary schools and its effects on health in children]. AB - PURPOSE: To elucidate the actual status of indoor air pollution at newly built or renovated elementary schools, and to evaluate its effects on health symptoms in the affected children. METHODS: In the classrooms of four newly built or renovated elementary schools in Osaka Prefecture, indoor air levels of formaldehyde and volatile organic compounds (VOC) were measured immediately, 1 month, 3 months, 10 months and 22 months after the completion of the construction work. Also, questionnaire surveys regarding subjective symptoms of sick building syndrome were conducted before and after the renovation on the children who attended classes in the renovated rooms. RESULTS: In the newly built computer classroom, more formaldehyde was detected one month after the completion of the construction work, when computers and furniture were carried in, than immediately after the completion of the work. Then, during the summer season, even 10 months and 22 months after completion of the new building, formaldehyde above the guideline values was detected. In the renovated common classrooms, the formaldehyde level was the same as that in the classrooms which did not undergo renovation, but VOC levels were higher immediately after the completion of the construction work, and the toluene level was above the guideline value. In 4 story reinforced concrete school buildings, indoor air pollution tended to be higher on the third and the fourth floors than on the first and the second floors. In 3-story school buildings, indoor air pollution tended to be higher on the third floor than on the second floor. The survey of subjective symptoms of the children revealed a tendency toward an increase in the prevalence of sick building syndrome after a renovation. However, the actual number of the children complaining of the symptoms hardly changed. Instead, the number of symptoms for each subject increased, and this increase was significant in 5th and 6th grade boys. CONCLUSION: In the some classrooms of newly built or renovated elementary schools, chemical substances above the guideline values may be detected. In such classrooms, more ventilation is required. PMID- 16218413 TI - [Safety awareness of anticancer drug handling among Japanese nurses]. AB - OBJECTIVES: The purpose of this study was to investigate Japanese nurses' awareness of the adverse effects of occupational exposure to anticancer drugs and safe handling, as well as their associations. METHODS: A self-rating questionnaire was mailed to 939 nurses in 107 university hospitals, 13 cancer special hospitals, and 193 general hospitals with over 300 beds and at least five or more clinics in Japan. A total of 571 female nurses responded to the questionnaire. RESULTS: About 40% of the nurses were not at all aware of the potential adverse effects of occupational exposure to anticancer drugs. Eighty eight percent of the nurses prepared anticancer drugs in the hospital wards; in most cases, nurses, not doctors or pharmacists, transported and administered such drugs to cancer patients. Regarding safety, 39% of the nurses took protective countermeasures against anticancer drugs; 15% paid special attention to the handling environment; 10% had guidelines for the handling of anticancer drugs; and, only 7% took countermeasures with body fluids or linen handling of cancer patients. Although 82% of the nurses were concerned about the potential health effects of occupational exposure, 75% or over did not know whether the exposure might affect their future health or progeny. Awareness of adverse effects was significantly related with precaution in anticancer drug handling, for which 95% of the nurses stated a desire for special education and training. CONCLUSIONS: Approximately 60% of Japanese nurses are aware of the risk of occupational exposure to anticancer drugs, but this might be an overestimate because of the sampling bias. Since only small numbers of nurses take sufficient safety precaution, programs for education and training for safe handling of anticancer drugs are crucial. PMID- 16218414 TI - [Significance of RFLP analysis for community-based control against tuberculosis]. AB - PURPOSE: The purpose of this study is to obtain an index for measures against tuberculosis by elucidation of the source/route of infections and prevention of secondary infection by RFLP analysis in Okayama. METHODS: A total of 474 strains isolated in Okayama prefecture from April 2000 to March 2004 were subjected to RFLP analysis. Analysis was only performed for mycobacterial strains from patients whose ages and places of residence were known. The prevalent situation of tuberculosis was analyzed by the distribution of IS6110 copy numbers, and the prevalent strains were examined by cluster analysis of RFLP patterns. To speculate regarding the mode of infections, annual change of RFLP patterns and differences in RFLP patterns with the age group and area were examined. A database was made with the results of RFLP analysis and information about bacterial strains from a public health center and monitored in order to discover latent links to outbreak or sporadic cases. RESULTS: The RFLP patterns for the 474 strains reflected past tuberculosis in highly prevalent times. Among 37 sets of clusters which showed the same pattern (110 strains), relationships were recognized between the patients in 20%. Most of the patients were younger than 60 years of age. By classification according to the similarity of RFLP patterns, three prevalent strain groups were recognized, to which about 40% of all strains belonged. Judging from the RFLP pattern distribution, there was no obvious annual change. The age groups of the patients and the areas where they lived did not influence the RFLP patterns of mycobacteria. Therefore, it was concluded that infection of the various age groups occurred with mycobacteria from reactivated older generation. Accordingly, prevention of tuberculosis reactivation in the aged can be considered the most important measure against tuberculosis. CONCLUSIONS: We obtained basic data on the prevalence for tuberculosis and the prevalent strains in Okayama by RFLP analysis. Together with conventional epidemiological investigation the resulting database allows scientific analysis of cases, although latent links to outbreak or sporadic cases were not revealed in this study. Further analyses with the database should contribute to measures against tuberculosis. PMID- 16218415 TI - [Survey on public health nursing education-in the comparison of nursing education courses, universities, advanced courses for public health nurse with junior nursing colleges, and public health nursing school]. AB - PURPOSE: Changes in public health nursing education have been consideration. Theses changes include a dramatic increase in the number of public health nurses (PHNs) who have enrolled for nursing courses at university. This study was conducted to assess the current status and future of public health nursing education as perceived by teachers and students at three types of schools: universities offering nursing courses, advanced courses for PHNs with junior nursing colleges, and public health nursing schools. METHODS: Questionnaires were distributed to teachers and students by mail. The questions that were sent to teachers asked which subjects were required to become a certified PHN, which lecture methods were employed to teach public health-particularly community health assessment methods, and what was the level of awareness of the activities of PHNs. Students were asked about their motivation to be a PHN, their understanding of public health, their views of public health activities and their images of PHNs. RESULTS: Responses were analyzed and differences between questionnaires from different schools were noted. These included the number of subjects and the total number of hours spent doing practical training and field experience in universities and the other types of schools, and the number of teachers. Differences also were noted among students at three types of schools about their age, methods of public health activities, knowledge about activities undertaken by PHNs, and their images of PHNs. No differences were observed among the schools with respect to the students' conceptual understanding of public health. CONCLUSION: Student age, practical training and field experience were found to contribute to their level of understanding of public health and public health nursing. It is thus necessary to consider the teaching methods employed by universities that administer nursing courses and the effectiveness of courses offered by graduate schools. PMID- 16218416 TI - [Q fever in acute exacerbation of chronic lower respiratory tract infection]. AB - We studied the effect of Q fever in acute exacerbation of chronic lower respiratory tract infection. The subjects consisted of 80 cases with acute exacerbation of chronic lower respiratory tract infection treated during the period from March 2002 till October 2004. Q fever was diagnosed using a PanBio Coxiella burnetii ELISA test kit. Two cases (2.5%) were positive for IgM in the acute stage, and were diagnosed as having acute infection by C. burnetii. They were elderly women with bronchiectasis, aged 76 and 82. They had no history of keeping cats or dogs, but the onset of acute exacerbation of chronic lower respiratory tract infection was June and March which is the breeding seasons for cats and dogs. Acute exacerbation of chronic lower respiratory tract infection were considerd to be a mixed infection with Pseudomonas aeruginosa (the 76-year case) and Haemophilus influenzae (the 82-year-case). It is concluded that C. burnetii can induce exacerbation of chronic lower respiratory tract infection, their cases were considerd to be mixed infection with C. burnetii and other bacteria. PMID- 16218417 TI - [An oxygen delivery device with a jet nebulizer function (Inspiron Nebulizer or Aquapak Nebulizer) can not provide high concentrations of oxygen to adult patients with respiratory failure]. AB - Oxygen delivery devices with a jet nebulizer function are widely used in Japan. The device, utilizing the Bernoulli principle, can provide oxygen concentrations of 24-100%. The present study demonstrated that maximal inspired oxygen concentration provided by this oxygen delivery device is theoretically 60%, as total gas flow is insufficient to meet all inspiratory requirements to provide oxygen concentrations of greater than 60%. This theoretical calculation was verified using a face model to which oxygen was delivered using an Inspiron Nebulizer or Aquapak Nebulizer. PMID- 16218418 TI - [A case of yellow nail syndrome associated with eosinophilic bronchial disease successfully treated with clarithromycin and budesonide]. AB - A 78-year-old man was referred to our hospital complaining of chronic productive cough. Physical examination revealed yellowish, thin nails and pretibial edema. A chest computed tomograph showed bilateral bronchiectasis. A sinus radiograph showed the findings of chronic sinusitis. From these findings, yellow nail syndrome was diagnosed. Long-term low-dose macrolide therapy with 400 mg/day clarithromycin was started and his symptoms began to gradually improve. However, complete resolution of his symptoms was not achieved and fiberoptic bronchoscopy was performed. Transbronchial biopsy specimen obtained from the right second carina showed bronchial asthma-like findings such as eosinophilic infiltration, thickening of the basement membrane, mucosal edema and goblet cell hyperplasia. Airflow reversibility was not detected. Thus a diagnosis of coexistence of yellow nail syndrome and eosinohilic bronchial disease was established. Further improvement of his symptoms was achieved by additional therapy with 800 microg/day budesonide and 100 microg/day salmeterol. To the best of our knowledge, this is the first report of a case of yellow nail syndrome associated with eosinophilic bronchial disease successfully treated with long-term low-dose macrolides and inhaled corticosteroids. PMID- 16218420 TI - [A case of mesalazine-induced pleuritis with hemophagocytic findings]. AB - A 25-year-old woman who had been receiving mesalazine for 2 weeks for ulcerative colitis presented with a nonproductive cough, high fever, and exertional dyspnea. Her chest radiograph showed bilateral pleural effusion. At first, infectious pleuritis was suspected and antibiotics were administered, but the pleural effusion increased and high fever continued. Because clinical and radiographic abnormalities markedly improved with discontinuation of mesalazine and treatment with corticosteroid, mesalazine was thought to be the cause of pleural effusion in this case. This case was thought to be of interest when we consider the onset mechanism of pleurisy by mesalazine. Mesalazine-induced pleuritis must be considered in patients who develop unexplained respiratory symptoms while taking this agent. PMID- 16218419 TI - [A case of bilateral diaphragmatic paralysis clinically suggestive of brachial neuritis]. AB - A 55-year-old man complained of acute onset of shoulder pain and dyspnea in the supine position. A diagnosis of bilateral diaphragmatic paralysis was made based on clinical inspection of his breathing pattern, radiographic appearance, and pulmonary function tests. He had had no traumatic or thoracic surgery or inflammatory episode. He did not suffer from diabetes, other central neural diseases, or any neoplastic disease. From the clinical feature and electromyographic findings, phrenic nerve involvement of brachial neuritis without any other muscle involvement was considered as a causative disease. When he received non-invasive intermittent positive-pressure ventilation by nasal mask in a supine position, his dyspnea was substantially attenuated and Carbon dioxide retention was lessened. After 1 year, his shoulder pain is still persisting and radiographic findings are not remarkably improved. PMID- 16218421 TI - [Stromal bone formation by lung adenocarcinoma]. AB - We report a case of lung adenocarcinoma with stromal bone formation. A 73-year old woman was incidentally found to have a coin lesion in the left lower lung field on a chest roentgenogram and computed tomography showed a nodular lesion containing a few coarse high density areas in the left lower lobe. Since transbronchial cytology revealed adenocarcinoma, left lower lobectomy was performed. Histologically, the tumor was a papillary-tubular adenocarcinoma, and fragments of osseous tissue were found within abundant fibrous stroma. Bone formation in primary lung adenocarcinoma is a very rare condition, and only 7 other cases have been reported in the literature. PMID- 16218422 TI - [Investigation of a pulmonary tuberculosis outbreak]. AB - Diagnoses of infectious tuberculosis (TB) patients were followed by thorough contact investigation on the basis of our hospital's Infectious Disease Manual ever since an infection of an inpatient with extended hospital stay was confirmed by a positive acid-fast sputum smear in October 1998. In September 2000, a nurse was found to have pulmonary TB and another was given a diagnosis of right tuberculous pleuritis the following November. Contact investigations were expedited among all hospital staff, families of the infected nurses, and all suspected inpatients. Five were diagnosed as TB, 8 were given chemoprophylaxis and 8 others required observation. The result verified a TB outbreak within the hospital, and management of TB infection control was re-enforced subsequently. We concluded that immediate contact investigation promoted successful early diagnosis, and reacknowledged the significance of the health supervision of all staff, operations including the environment and equipment control of the institution, and frequent contact and integration with the administration of the public health center. This experience enabled a useful revision of the disease manual for the future. PMID- 16218423 TI - [A case of extramedullary plasmacytoma in the mediastinum]. AB - A 80-year-old man was admitted to our hospital for examination of a mediastinal mass. Chest CT showed a 6 x 9 cm mass in the left anterior mediastinum. The left main bronchus and pulmonary artery were invaded by the mass. Bronchoscopic examination showed compression of the left main bronchus by the mass and enlarged lymph nodes. The left upper lobe and lingular bronchi were almost occluded. The overlying mucosa was invaded by tumor tissue. Microscopic findings of specimen obtained by transtracheal biopsy showed plasma cells of various sizes were at different stages of differentiation. Immunohistochemical staining revealed IgA monoclonality. Whole-body bone scan and bone marrow aspiration excluded multiple myeloma and a diagnosis of extramedullary plasmacytoma was made. Serum protein electrophoresis showed an IgA monoclonal gammopathy. Immunoelectrophoresis of the serum revealed an IgA kappa monoclonal component. These suggested that tumor cells were producing monoclomal IgA kappa light chain. Radiotherapy was effective in reducing the size of the plasmacytoma, and serum IgA reverted to normal. A 67 Ga scan showed strongly increased uptake in the left anterior mediastinum. Chest CT after radiotherapy showed that the decreased mass had an extrapleural sign against the mediastinal pleura. We determined that the origin of the plasmacytoma was probably the mediastinum. We reported a rare case of extramedullary plasmacytoma in the mediastinum. PMID- 16218424 TI - [Clinical features of intradural spinal cord metastases in primary lung cancer]. AB - We retrospectively investigated the clinical features of 9 consecutive cases of intradural spinal cord metastasis from primary lung cancer treated at our hospital between April 1999 and March 2002. During those three years, spinal cord metastasis was diagnosed in seven of 49 (14.3%) cases with small cell carcinoma and only two of 284 (0.7%) cases with non-small cell carcinoma. Eight of the 9 cases had concomitant brain metastasis that preceded spinal cord metastasis and had received brain irradiation. The other patient without brain metastasis had also received prophylactic cranial irradiation. The interval from brain irradiation to the diagnosis of spinal cord metastasis ranged from 116 to 708 days (median 183 days). The most common initial symptom was muscle weakness of the lower extremities in five cases. Seven of the patients rapidly developed transverse myelopathy within two weeks. Magnetic resonance imaging (MRI) with contrast enhancement demonstrated intramedullary tumors in seven cases and intradural extramedullary tumors in two cases. Spinal cord metastasis was often multifocal, and in each case lumbar enlargement was commonly involved. Radiation therapy with or without concurrent chemotherapy produced both neurologic improvement and a tumor response shown on MRI in 4 patients. Heightened awareness of the increasing incidence and background risk factors of this unusual complication could lead to earlier diagnosis and more effective treatment for neurologic palliation. PMID- 16218425 TI - [A case of Wegener's granulomatosis without PR3-ANCA at relapse]. AB - A 49-year-old man was admitted to our hospital with fever. His chest radiograph showed some nodules in the right upper and lower lung fields. The cytoplasmic antineutrophil cytoplasmic antibody test was positive, and histopathologic biopsy of a small nasal polyps yielded a diagnosis of Wegener's granulomatosis. He was started on prednisolone and cyclophosphamide. The findings on his chest radiograph and his symptoms improved rapidly, and we stopped these drugs after one year. Two years after cessation of treatment, his chest radiograph showed two nodules with cavities. Relapse of Wegener's granulomatosis was diagnosed. The proteinase 3-antineutrophil cytoplasmic antibody test was negative. He was started on prednisolone and cyclophosphamide, and the findings on his chest radiograph improved rapidly. Chest radiographs are useful for follow-up observation of patients with Wegener's granulomatosis after treatment. PMID- 16218426 TI - [Impact of metabolic syndrome and diabetes mellitus on cardiovascular events in coronary artery disease without ischemia on stress thallium-201 single photon emission computed tomography after percutaneous coronary intervention]. AB - OBJECTIVES: The metabolic syndrome defined by the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) is a predictor of cardiovascular events. However, the significance of metabolic syndrome for cardiovascular events has been not clarified in Japan. The impact of metabolic syndrome and diabetes mellitus on cardiovascular events was investigated, especially in the high risk group after percutaneous coronary intervention. METHODS: We studied 456 patients (mean age 63 +/- 10 years, range 36-88 years) without ischemia on stress thallium 201 single photon emission computed tomography after percutaneous coronary intervention. The diagnosis of metabolic syndrome was made according to the modified NCEP ATP III criteria. Cardiovascular events were examined for mean 3.7 +/- 1.8 years (range 2.0-8.7 years). There were 196 patients without diabetes mellitus or metabolic syndrome (Group D - M -), 89 patients without diabetes mellitus but with metabolic syndrome (Group D - M +), 61 patients with diabetes mellitus but without metabolic syndrome (Group D + M -), and 110 patients with both diabetes mellitus and metabolic syndrome (Group D + M +). RESULTS: The event free survival curve in Group D - M + was significantly lower than that in Group D - M - (p < 0.05), but not different from that in Group D + M -. The survival curve was markedly lower in Group D + M + than that in Group D - M + (p < 0.005). The Cox proportional hazard model revealed that diabetes mellitus and metabolic syndrome were independent significant risk factors for events. CONCLUSIONS: The diagnosis of metabolic syndrome was helpful for identification of patients with high cardiovascular event rate even in patients after percutaneous coronary intervention. The combination of metabolic syndrome and diabetes mellitus markedly increases the risk for cardiovascular events. PMID- 16218427 TI - [Ultrasonographic assessment of carotid atherosclerosis for the differentiation of ischemic cardiomyopathy and dilated cardiomyopathy]. AB - OBJECTIVES: We assessed the clinical value of carotid intima-media thickness (IMT) for the differentiation of ischemic cardiomyopathy (ICM) and dilated cardiomyopathy (DCM). METHODS: We studied 38 patients who showed left ventricular dilation and low ejection fraction (< 40%) by cardiac catheterization. There were 15 patients with ICM which was defined as stenosis of > 75% at one major branch or more. Twenty-three patients were found to have DCM as diagnosed by histological examination using biopsy specimens. Carotid IMT visualized by B-mode imaging was measured at 16 sites in the extracranial carotid arteries, and the maximum IMT, mean IMT, and plaque score were calculated. These parameters together with cardiovascular risk factors including age, sex, hypertension, diabetes mellitus, hypercholesterolemia, cigarette smoking, and obesity were compared between the ICM and the DCM groups. RESULTS: The maximum IMT, mean IMT, and plaque score in the ICM group were 2.80 +/- 1.63 mm, 1.21 +/- 0.36 mm, 13.05 +/- 8.12, respectively, which were significantly higher than the corresponding parameters in the DCM group (1.19 +/- 0.51 mm, 0.73 +/- 0.15 mm, 1.52 +/- 2.51; p < 0.001, respectively). Furthermore, plaque score of greater than 5 was found to be an excellent diagnostic parameter for ICM with 91% sensitivity and 100% specificity. The cardiovascular risk factors did not differ between the two groups. CONCLUSIONS: Our observations demonstrate that assessment of carotid IMT is a clinically useful tool to differentiate ICM and DCM in patients with left ventricular dysfunction. In particular, coronary angiography should be recommended in patients with a plaque score of greater than 5. PMID- 16218428 TI - [Hypoadiponectinemia implies the development of atherosclerosis in carotid and coronary arteries]. AB - OBJECTIVES: Adiponectin is an adipocyte-derived endocrine factor. Hypoadiponectinemia has been observed in obese patients, and plasma adiponectin levels are reported to increase during weight reduction. Moreover, hypoadiponectinemia has also been observed in patients with coronary artery diseases. The present study investigated the relationships between levels of adiponectin and carotid intimal-medial thickness, a marker of early vascular disease, and carotid artery plaque and the severity of coronary artery disease, a marker of advanced vascular disease. METHODS: Four hundred thirty-one consecutive patients were enrolled from inpatients without acute coronary syndrome who underwent coronary angiography between August 2001 and August 2003. The residual adiponectin levels were calculated by adjusting for sex, age, and body mass index, and a logarithmic transformation was applied. The severity of coronary artery disease was evaluated by coronary angiography and divided into four groups (Group 0: no significant organic stenosis, Group 1: 1-vessel disease, Group 2: 2 vessel disease, Group 3: 3-vessel disease or left main coronary trunk disease). Carotid plaque was evaluated by ultrasonography and divided into two groups [Group(-) : patients without carotid plaque, Group (+): patients with carotid plaque]. The intimal-medial thickness was measured on a longitudinal scan of the common carotid artery at a point 1 cm proximal from the bifurcation bulb. RESULTS: The logarithmic-transformed levels of residual adiponectin were associated with severity of coronary artery disease (Group 0: 0.18 +/- 0.59 microg/ml, Group 1: -0.02 +/- 0.56 microg/ml, Group 2: - 0.09 +/- 0.58 microg/ml, Group 3: - 0.10 +/- 0.66 microg/ml, p = 0.0013). The logarithmic-transformed levels of residual adiponectin were decreased in patients with carotid plaque [Group (-): 0.08 +/- 0.59 microg/ml, Group (+): - 0.08 +/- 0.59 microg/ml, p = 0.045]. However, the logarithmic-transformed levels of residual adiponectin were not associated with intimal-medial thickness (p = 0.6398). CONCLUSIONS: Hypoadiponectinemia adjusted for sex, age, body mass index implies the progression of carotid and coronary sclerosis. PMID- 16218429 TI - Long-term follow-up of a patient with Kawasaki disease and coronary aneurysm associated with asymptomatic thrombosis: a case report. AB - A 20-year-old male was first diagnosed with Kawasaki disease at age 2 years 9 months. Coronary angiography in the acute phase revealed coronary aneurysms, so chronic antiplatelet therapy was initiated with aspirin and ticlopidine. The patient was asymptomatic and was followed up. Stress myocardial imaging showed asymptomatic myocardial ischemia at age 20 years. Coronary angiography was performed, and revealed 99% occlusion of the right coronary artery and collateral circulation from the left coronary artery. Occlusion was attributed to coronary aneurysm thrombosis. Much remains unknown about the long-term prognosis in patients with coronary aneurysm associated with Kawasaki disease. Asymptomatic children who are followed up sometimes develop ischemic heart disease as young adults. This case highlights the need for long-term follow-up in patients with Kawasaki disease and coronary aneurysms. PMID- 16218430 TI - [Aortic stenosis and mitral regurgitation complicated by hemolytic anemia and positive Direct Coombs test: a case report]. AB - A 83-year-old man was admitted because of heart failure due to severe aortic stenosis and mitral regurgitation secondary to chordal rupture of the anterior leaflet. Mild anemia and elevated serum lactate dehydrogenase were present with reticulocytosis and haptoglobinemia. Direct Coombs test was positive. Coexistence of autoimmune hemolytic anemia was identified, but the main cause of his hemolysis was thought to be mechanical hemolysis due to stenotic valve and/or ruptured chordae because of the presence of red cell fragmentation. The patient successfully underwent double valve replacement. Improvement of anemia was coupled with reduction of the serum lactate dehydrogenase level. Valvular shear stress on the red cells and reduction of red cell deformability secondary to autoimmune hemolytic anemia were thought to be responsible for his hemolysis. PMID- 16218431 TI - [Rare cause of severe mitral regurgitation]. PMID- 16218432 TI - [Treatments of diabetic retinopathy in the future]. PMID- 16218433 TI - [Intraocular penetration of cefotiam hydrochloride, a kind of cephalosporin after filtration surgery in rabbit eyes]. AB - PURPOSE: In this study, we measured the cefotiam dihydrochloride (CTM) concentration in ocular tissue after filtration surgery in rabbit eyes. METHODS: CTM (20 mg/kg body weight) was administered intravenously 30 min before filtration surgery which was performed by double flap procedure on the right eyes of white rabbits. The aqueous humor and serum were extracted at 10 min after surgery and at 30 min, 60 min, and 120 min. Drug concentration in all of the specimens was measured by high performance liquid chromatography (HP-LC). RESULT: The CTM concentrations of aqueous humor in the nonoperated eyes were 0.44 +/- 0.16(mean +/- standard deviation) microg/ml (n = 4) (40 min after intravenous dosage), 0.36 +/- 0.17microg/ml (n = 4) (60 min after intravenous dosage), 0.38 +/- 0.34, microg/ml(n = 3) (90min after intravenous dosage) and 0.27 +/- 0.10 microg/ml (n = 5) (150 min after intravenous dosage). In contrast, CTM concentration in the aqueous humor of the operated eyes was 2.4 +/- 0.95 microg/ml (n = 4) at 10 min after surgery (40 min after intravenous dosage), 2.11 +/- 1.10 microg/ml (n = 4) at 30 min after surgery (60 min after intravenous dosage), 1.18 +/- 0.78 microg/ml (n = 4) at 60 min after surgery (90 min after intravenous dosage) and 0.47 +/- 0.1 microg/ml (n = 5) at 120 min after surgery (150 min after intravenous dosage). The intraocular penetration of CTM at 10 min and at 120 min after filtration surgery was significantly higher in comparison with the drug concentration in the nonoperated eyes (p < 0.05). CONCLUSION: The intraocular penetration of CTM after filtration surgery was much higher in comparison with the drug concentration in the nonoperated eyes. These results may be useful to predict the intraocular penetration of CTM in human eyes after filtration surgery. PMID- 16218434 TI - [Grading of diabetic retinopathy from non-stereoscopic color fundus photographs- relationship to fluorescein angiography findings and three-year prognosis]. AB - PURPOSE: To evaluate the grading of diabetic retinopathy from non-stereoscopic color fundus photographs, we examined the relation of the photos to fluorescein angiography findings and to the three-year prognosis. METHODS: Fifty diabetic patients(70 eyes) who had severe non-proliferative diabetic retinopathy or early proliferative retinopathy without photocoagulation treatment were graded regarding 11 items of four-field fundus photographs using a 50-degree mydriatic camera. Fluorescein angiography was performed and the relation of the images to the grades of diabetic retinopathy were analyzed. In 51 eyes, the relation between the grading and the progression and treatment of diabetic retinopathy were evaluated. RESULTS: The grading of microaneurysms and retinal hemorrhages (p < 0.001), soft exudates (p = 0.01), intraretinal microvascular abnormalities (p < 0.001), arteriolar white threads (p = 0.003), venous loops (p = 0.01), and new vessels (p < 0.001) was significantly related to the nonperfused areas on fluorescein angiograms. The grading of diabetic retinopathy was also significantly related to the threeyear prognosis. CONCLUSION: These results indicated that the grading of diabetic retinopathy from fundus photographs may have a potential advantage over conventional classifications of diabetic retinopathy. PMID- 16218435 TI - [Incidence of benign and malignant lesions of eyelid and conjunctival tumors]. AB - PURPOSE: To examine the incidence of benign and malignant eyelid lesions and conjunctival tumors. SUBJECTS AND METHODS: One-hundred-and twenty-eight cases (131 eyes) which were treated during the period from January 1990 to February 2004 were histopathologically diagnosed for eyelid or conjunctival tumors (87 cases of eyelid tumors and 41 cases of conjunctival tumors) in retrospective evaluations. The incidence of benign or malignant lesions, the pathological classification, age, sex, and clinical diagnostic accuracy were all investigated. RESULTS: Sixty-four (73%) of the tumors were found to be benign eyelid tumors. The common benign eyelid tumors were 14 nevocellular nevi, 9 seborrheic keratosis, 7 epidermoid cysts, and 6 papillomas. Twenty-four (27%) eyelid tumors were malignant. These included 9 basal cell carcinomas, 9 sebaceous gland carcinomas, 4 malignant lymphomas, and 2 metastatic tumors. Thirty-four (79%) conjunctival tumors were benign, and the common benign conjunctival tumors were 9 nevocellular nevi and 7 papillomas. Nine (21%) conjunctival tumors were malignant, comprising 7 malignant lymphomas and 2 squamous cell carcinomas. The mean ages of malignant eyelid and conjunctival tumor patients were significantly older than those of benign tumor patients. Clinical accuracy in predicting basal cell carcinoma and sebaceous gland carcinoma was 11.1% and 44.4%, respectively. CONCLUSIONS: Approximately 70 approximately 80% of all eyelid and conjunctival tumors are benign. Clinicians should suspect that the lesions are malignant when seeing elderly patients with eyelid or conjunctival tumors. Excised eyelid lesions should be submitted for histopathologic confirmation because there are some cases where clinical diagnosis does not match pathological diagnosis. PMID- 16218436 TI - [Clinical results of amniotic membrane transplantation in Miyata Eye Hospital according to surgical methods]. AB - PURPOSE: To evaluate clinical results of amniotic membrane transplantation according to three methods : graft, patching, and stuffing. SUBJECT AND METHODS: We retrospectively reviewed 65 patients(65 eyes) who received amniotic membrane transplantation by the three methods, graft, patching or stuffing during the period of March 1998 to May 2003 in the Miyata Eye Hospital. Application of surgery was determined as follows : graft with limbal transplantation for limbal dysfunction, patching for prolonged epithelial defect, and stuffing for corneal perforation. The criteria of success were defined as corneal and conjunctival epithelialization and prevention of invasion of proliferative tissues beyond the limbus for graft, corneal epithelialization for patching, and occlusion of perforated cornea at first use for stuffing, respectively. PMID- 16218437 TI - [Preoperative factors for postoperative resolution of metamorphopsia in idiopathic macular hole surgery]. AB - PURPOSE: To elucidate factors for resolution of metamorphopsia after successful surgery for idiopathic macular hole. PATIENTS AND METHODS: A series of 15 cases of 15 patients with successful surgery for idiopathic macular hole were evaluated for signs of metamorphopsia before and one year after surgery. Preoperative and postoperative assessments included testing of metamorphopsia using M-CHART and optical coherence tomography. The postoperative state of the metamorphopsia was judged to have resolved when the test score was 0.2 or less and to be residual when it was more than 0.2. Classification of preoperative factors in two groups of patients included evaluation of metamorphopsia score, visual acuity, optical coherence tomography findings, and clinical stage of macular hole. RESULTS: Six of 15 eyes (40%) showed postoperative resolution of metamorphopsia. There was no significant correlation of postoperative metamorphopsia score with preoperative score or preoperative visual acuity. Eyes with postoperative resolution of metamorphopsia had a macular hole with a significantly smaller diameter and a significantly thinner retina surrounding the hole. Two eyes with clinical stage 4 macular hole had residual metamorphopsia after surgery. CONCLUSIONS: The diameter of a macular hole and the thickness of the retina surrounding a macular hole are significant preoperative factors for postoperative resolution of metamorphopsia after idiopathic macular hole surgery. PMID- 16218438 TI - [Role of chemokines in the vitreous of proliferative diabetic retinopathy]. AB - PURPOSE: To investigate the role of chemokines in the pathogenesis of proliferative diabetic retinopathy (PDR). SUBJECTS AND METHODS: In total, 41 eyes of 38 patients undergoing vitrectomy were divided into two groups; PDR and non PDR. The PDR group was comprised of 30 eyes, and the non-PDR group of 11 eyes. Vitreous specimens obtained at vitrectomy were centrifuged and separated into supernatants and cellular components. Concentrations of vascular endothelial growth factor(VEGF), interleukin-8 (IL-8), monocyte chemotactic protein-1 (MCP 1), and regulated upon activation, normal T cell expressed and secreted(RANTES) in the supernatants were determined by enzyme-linked immunosorbent assay(ELISA) or chemiluminescence enzyme immunoassay(CLEIA). Expression of VEGF in the cellular components was determined by immunohistochemistry. RESULTS: Vitreous levels of VEGF(p < 0.05), IL8 (p < 0.0001) and MCP-1 (p < 0.05) in the PDR group were significantly higher than in the non-PDR group. However, there was no significant difference in RANTES between the two groups. There was a significant correlation (p < 0.0001, r = 0.84) between vitreous IL-8 and MCP-1 levels in the PDR group. After immunohistochemical staining with antiVEGF monoclonal antibody, VEGF positivity was localized in polymorphonuclear leukocytes and monocytes of the cellular components of PDR vitreous specimens. CONCLUSIONS: These results indicate that chemokines are possibly involved in the recruitment of neutrophils and monocytes into the vitreous and that they play a role in the intraocular neovascularization characteristic of PDR. PMID- 16218439 TI - [Recent outcomes of vitreous surgery for diabetic retinopathy]. AB - PURPOSE: To investigate background, surgical method, complications, prognosis, and prognostic factors in patients undergoing vitrectomy for diabetic retinopathy. SUBJECTS AND METHODS: Three hundred and forty eyes of 261 patients undergoing vitrectomy for diabetic retinopathy in five recent years were studied regarding background, surgical method, complications, and visual prognosis. Factors influencing postoperative visual acuity and complications were also examined using univariate and multivariate analyses. RESULTS: Final postoperative visual acuity (FPVA) improved in 226 eyes (66%). FPVA of 0.1 or better and 0.5 or better was achieved in 80% and 45% of all patients, respectively. Postoperative complications occurred in 89 eyes(26%). In the vitreous hemorrhage group, FPVA improved in 86%, and FPVA of 0.5 or better was achieved in 60%. Postoperative complications were most common in the traction detachment group and the percentage was 40%. Factors influencing FPVA were preoperative visual acuity, postoperative complications, indications for surgery, and preoperative severity. Factors influencing postoperative complications were patient background, preoperative visual acuity, preoperative severity, and iatrogenic breaks. CONCLUSIONS: Vitrectomy is a useful method for diabetic retinopathy but postoperative complications must be managed. PMID- 16218440 TI - [Dramatic alteration in optic nerve head topography after filtering surgery in an adult patient with traumatic glaucoma]. AB - PURPOSE: We treated an adult patient with traumatic glaucoma who exhibited dramatic elasticity in the topography of the optic nerve head, as shown in a Heidelberg Retina Tomograph (HRT) examination after filtering surgery. CASE: A 41 year-old man came to us with a right eye injury following an external blow. At the first examination, visual acuity was 0.4 (1.2) and intraocular pressure (IOP) was 11 mmHg in both eyes. Cells in the anterior chamber and angle dialysis were noted in all quadrants of the right eye, but there was no hyphema. Two weeks later, the patient returned to our hospital with pain in the right eye and a headache. Visual acuity in the right eye had decreased to 0.06 (0.1) and IOP was 50mmHg. A cilioconjunctival injection and corneal edema were observed. The patient was treated with medications, but IOP was uncontrollable and the optic disc cupping in the right eye was enlarged. Further, values for Cup Disc Area Ratio (C/D; 0.553) and Cup Volume (CV; 0.548) in the right eye were larger than those of the left (0.287 and 0.168, respectively) in an HRT examination. We performed a non-penetrating trabeculectomy 5 weeks after the injury. Two weeks after surgery, IOP was lowered to 7 mmHg, and C/D 0.122) and CV (0.062) were improved. Six months after the procedure, C/D (0.304) and CV (0.202) were nearly the same as those of the left eye (0.292 and 0.144). CONCLUSION: Although the duration of high IOP was short, high elasticity in the optic disc morphology of this adult case was shown quantitatively in an HRT examination. PMID- 16218441 TI - [Isolated sphenoid sinus lesions: a clinical analysis of 44 cases]. AB - OBJECTIVE: We reviewed isolated sphenoid sinus lesion (ISL), a relatively uncommon entity found mostly in departments other than otolaryngology. STUDY DESIGN: We retrospectively studied 44 cases of ISL at the Department of Otolaryngology, Kagoshima City Hospital, Kagoshima, Japan, for the 5 years between 1999 and 2003. METHODS: Data was obtained for each patient regarding age, present symptoms, first department visit, pathological classification, and therapy. RESULTS: Eighty percent of patients first visited departments other than otolaryngology. Of these, 32 had inflammatory disease, 8 mucocele, 1 a benign tumor, and 3 malignant tumors. The most common symptoms were headache at 59% followed by ocular symptoms at 27%. Ocular symptoms included ocular pain in 60% of those with inflammatory disease, visual disturbance in 63% of those with mucocele, and diplopia in all of those with neoplasms. CONCLUSION: The type of ocular symptom largely depends on ISL pathology. Patients with headaches and diplopia have a high ratio of malignant neoplasms. Endoscopic sinus surgery was useful in diagnosing and treating ISL in cases in which conservative therapy was ineffective. It is most important to differentiate pathological disease that requires early diagnosis and therapy such as acute inflammatory disease, mucocele with visual disturbance, and malignant tumors. PMID- 16218443 TI - [A rare case of ectopic, normally functioning thyroid tissue presenting as a left submandibular mass]. AB - This report describes the case of a 53-year-old female, who presented with a painless swelling in the left submandibular region which had gradually increased in size. Thirty years ago, this patient underwent subtotal thyroidectomy at another hospital and her thyroid function subsequently appeared normal. She presented no symptoms of hypothyroidism when she visited our hospital. A clinical diagnosis of a benign tumor arising from the submandibular gland was made with enhanced CT scan, MRI and technesium scintigraphy. The submandibular mass was surgically excised and the resected tissue revealed a well-delineated, encapsulated, solid and reddish-brown colored mass. Histological examination confirmed ectopic thyroid tissue with partial adenomatous goiter including vacuolated rich colloid in the thyroid follicles which suggested compensatory hyperplasia. Because this patient presented with post-operative hypothyroidism, we concluded that this ectopic lesion had continued to secrete thyroid hormone. Ectopic thyroid tissue should be considered in the differential diagnosis of swellings involving the submandibular area, especially if the ectopic tissue would be the only functioning thyroid tissue. PMID- 16218442 TI - [Incidence of vertigo and dizziness disorders at a university hospital]. AB - Subjects were 626 patients reporting vertigo or dizziness seen at the University Hospital Department of Otolaryngology from April 2001 to September 2003. Patients were diagnosed based on diagnostic criteria prescribed by the Japan Society for Equilibrium Research. The most common peripheral vestibular disorder was benign paroxysmal positional vertigo (32%), followed by Meniere's disease (12%). All peripheral vestibular disorders accounted for 65%. Central vestibular disorder accounted for 7%, of which space-occupying lesions in the posterior fossa accounted for 1.0% and cerebral infarction 1.9%. Dizziness due to orthostatic hypotension accounted for 4.0%. Among past reports on clinical statistics of vertigo, the incidence of vertigo and dizziness disorders differed greatly, but our research and the past 2 reports based on diagnostic criteria prescribed by the Japan Society for Equilibrium Research showed almost the same incidence, i.e., BPPV of 30-40%, Meniere's disease of 7-10%, other peripheral vestibular disorders of 15-20%, and central vestibular disorder of 6-8%. Unified diagnostic criteria are thus important in the statistical analysis of vertigo disorders. PMID- 16218444 TI - [A case of tetanus showing limited mobility of the bilateral vocal cords]. AB - Patients with tetanus complaining of common signs such as lockjaw, voice disturbance, and difficulty in swallowing, etc, should never be forgotten. A 71 year old woman complained of dyspnea and difficulty in swallowing. She had lockjaw with tongue bite soon after a tracheostomy, and she was diagnosed as having tetanus. She showed the redundant gag reflex even to a slight stimulus and limited mobility of bilateral vocal cords, which improved following intravenous administration of diazepam. This case suggests that the mobility of the vocal cords and the movement of the larynx could be limited in tetanus because even a slight stimulus caused a strong gag reflex, resulting in dyspnea or difficulty in swallowing. PMID- 16218445 TI - [Detection of tuberculosis infection using a whole blood interferon gamma assay in a contact investigation--evaluation using quantiFERon TB-2G]. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of a novel method of detecting tuberculosis infection, QuantiFERON TB-2G (QFT), in a large scale contact investigation when an outbreak of mass tuberculosis infection was suspected. SUBJECTS AND METHODS: The index case was a health-care worker who worked in a maternity hospital. The investigated contacts were categorized as follow according to the grade of closeness of contact; the "very close" contact group (11 subjects), the "close" contact group (33 subjects), and the "non-close" contact group (3,791 subjects). For the former two groups, tuberculin skin test (TST), chest X-ray examination and QFT were conducted. For the last contact group, TST and chest X-ray examination were conducted only to subjects who aged less than 29 years old, while only chest X-ray examination was conducted to those aged 30 years or older. The QFT test, i.e., a whole blood interferon-gamma assay using Mycobacterium tuberculosis-specific antigens, was performed to the "very close" and "close" contacts, and for strong tuberculin reactors among the "non close" contacts. RESULTS: The number of infected subjects in the "very close" contact group, the "close" contact group, and the "non-close" contact group were 7, 7, and 277, respectively, based on TST results. On the other hand, the number of infected subjects in each group were 3, 2, and 5, respectively, based on the QFT test. CONCLUSION: If the indication of chemoprophylaxis was determined based on TST test, this case would have been regarded a large tuberculosis outbreak. However, the use of the QFT test greatly reduced the number of the infected persons, so that the possibility of such massive TB outbreak was denied. Thus, the use of QFT, with which TB infection could be detected more accurately, seems to be very beneficial in contact investigations. PMID- 16218446 TI - [Clinical characteristics in non-symptomatic infants with tuberculosis whose diagnostic clue was regular tuberculin skin test prior to BCG vaccination]. AB - OBJECTIVE: Tuberculin skin test (TST) is a useful tool to find non-symptomatic infants with tuberculosis. In 2005, Japanese government starts direct BCG vaccination without prior TST, and we may miss one of the chances to find the above patients. In this study, we tried to find if there is any clinical examination to find non-symptomatic infants with tuberculosis instead of regular TST. SUBJECTS AND METHODS: We found out 20 non-symptomatic infants with tuberculosis among 131 infants who referred to our clinic because of positive TST prior to BCG vaccination from 1996 to 2003. We examined the data of gastric aspirate examination for Mycobacterium tuberculosis, complete blood counts, and serum chemistries in the 20 infants with tuberculosis. We also tried to find out the source of tuberculosis of these patients by the contact investigation. RESULTS: Induration of TST among 20 non-symptomatic infants with tuberculosis were 7 to 26 (median, 13.5) mm. Mycobacterium tuberculosis from gastric aspirates was positive in 13 infants (65%). The level of blood hemoglobin was more than 11 g/dl in all infants. The white blood cell count was increased in 6 infants (30%), and exaggerated lymphocyte predominance was recognized in 8 infants (40%). Serum CRP was normal, or subtly increased. Serum GOT (AST) was increased in 15 infants (75%), though the reason was unknown. Serum albumin was normal in all the infants. In the contact investigation, suspected sources were found in 11 infants (55%), and 6 out of 11 sources (55%) were their mothers. Nine sources did not belong to the risk groups for tuberculosis except two mothers coming from tuberculosis epidemic countries. CONCLUSION: In 20 non-symptomatic infants with tuberculosis, examination of gastric aspirate for Mycobacterium tuberculosis, TST, and chest imaging were useful tools to diagnose tuberculosis. However, these examinations are not done in non-symptomatic infants. Eleven family members who might infect the infants were found by the contact investigations, and none of them realized that they have tuberculosis. Our study suggests that there is no realistic clue to find non-symptomatic infants with tuberculosis instead of regular TST, and that the control of infant's tuberculosis still depends on the examination of close contacts of sporadic adult patients with tuberculosis. PMID- 16218447 TI - [A case of pulmonary mycobacterium intracellulare infection complicated with pneumothorax and pleurisy]. AB - A 74 year-old female complaining of increased cough and sputum was admitted to our hospital on June 14th 2004. She had been diagnosed as Mycobacterium intracellulare (M. intracellurare) infection since 2002 and had been treated from March to October 2003 in the Department of General Medicine in our hospital. Chest CT on admission showed diffuse small nodular shadows in the lung, a cavity, pneumothorax, and pleural effusion in the right lung. The sputum smear was positive for acid-fast bacilli and sputum PCR examination was positive for M. intracellulare. She was diagnosed as the recurrence of non-tuberculous mycobacterium (NTM) infection and treatment of NTM infection was started. No other infections were suspected and the pneumothorax and pleural effusion gradually improved with the treatment. We concluded that the pneumothorax and pleural effusion were caused by NTM infection. Since pneumothorax is an extremely rare complication in NTM infections we thought it is worth-while to report our case. PMID- 16218448 TI - Tuberculosis: the global timebomb. PMID- 16218449 TI - [Replacing the tuberculin skin test with a specific blood test]. AB - For almost 100 years has the tuberculin skin test (TST) been used for the support the diagnosis of active and latent TB infection. The TST test has, however, a number of limitations most notable low specificity in BCG vaccinated individuals due to cross-reactive components in PPD and the M. bovis BCG vaccine strain and an intensive search for new and more specific diagnostic antigens has therefore be ongoing. In this review we describe the discovery process leading to the identification of the M. tuberculosis specific antigens ESAT6 and CFP10; two low molecular weight proteins which are highly sensitive and specific for detection of a M. tuberculosis infection. PMID- 16218450 TI - [Metabolic pathway, genes, and enzymes for the degradation of chlorinated pesticide, gamma-hexachlorocyclohexane]. PMID- 16218451 TI - [Diversity of component interactions in the bacterial Rieske non-haem iron oxygenase systems]. PMID- 16218452 TI - [Mobile genetic elements for microbial degradation of environmental pollutants]. PMID- 16218453 TI - [Structure function relationship of the LysR type transcriptional regulator]. PMID- 16218454 TI - [PCB degradation systems in Rhodococcus bacteria: multiplex enzyme system determined by multiple isozyme genes]. PMID- 16218455 TI - [Biochemical and genetic bases of chloroethene-dehalorespiring bacteria]. PMID- 16218457 TI - [Molecular mechanisms underlying sex-pheromone reception in insects]. PMID- 16218456 TI - [Development of site- and phosphorylation state-specific antibodies]. PMID- 16218458 TI - Heart, rheumatism and autoimmunity: an old intriguing link. PMID- 16218459 TI - Inflammatory heart disease: a role for cytokines. AB - Inflammatory heart disease is a rising concern worldwide. Similar mechanisms link autoimmune diseases, including the association of increased disease with proinflammatory cytokines and the importance of regulatory mechanisms in the control of chronic inflammation. Many pathogens including bacteria, protozoa and viruses have been associated with heart disease in patients, and are able to induce similar disease in animal models. Recognition of pathogens by the innate immune system leads to release of proinflammatory cytokines that both reduce infection and increase chronic inflammatory heart disease. Elevated levels of proinflammatory cytokines are able to overcome tolerance to chronic disease, indicating that environmental factors are important in determining progression to chronic heart disease. Understanding the mechanisms leading to chronic heart disease will be critical for developing effective therapies to reduce cardiac dysfunction and heart failure. PMID- 16218460 TI - Autoimmune myocarditis and dilated cardiomyopathy: focus on cardiac autoantibodies. AB - Criteria of organ-specific autoimmunity are fulfilled in a subset of patients with myocarditis/dilated cardiomyopathy (DCM). In particular, circulating heart reactive autoantibodies are found in patients and symptom-free relatives. These autoantibodies are directed against multiple antigens, some of which are expressed in the heart (organ-specific), others in heart and some skeletal muscle fibres (partially-heart specific) or in heart and skeletal muscle (muscle specific). Distinct autoantibodies have different frequency in disease and normal controls. Different techniques detect one or more antibodies, thus they cannot be used interchangeably for screening. It is unknown whether the same patients produce more antibodies or different patient groups develop autoimmunity to distinct antigens. IgG antibodies, shown to be cardiac and disease-specific for myocarditis/DCM, can be used as autoimmune markers for identifying patients in whom immunosuppression may be beneficial and relatives at risk. Some autoantibodies may also have a functional role, but further work is needed. PMID- 16218461 TI - Nonorgan specific autoantibodies and heart damage. AB - Heart damage, mediated by different autoantibodies can involve several anatomical heart structures: valves, arteries, conduction tissue. Verrucous endocarditis is frequently reported in patients with antiphospholipid syndrome (APS) with or without systemic lupus erythematosus (SLE), particularly if they suffer from central nervous system involvement. Antiphospholipid antibodies (aPL) were shown deposited at subendothelial level of the affected valves. According to several in vitro and in vivo experimental models, aPL, anti-oxidized LDL (oxLDL), anti-heat shock protein 65 (HSP65) and anti-endothelial cells antibodies (AECA) seem to be involved in the pathogenesis of the atherosclerosis phenomena described in systemic autoimmune disease and vasculitis. However, the observation of the association of the same antibodies with clinical and subclinical atherosclerosis in patients is still controversial. The children of anti-Ro/SSA positive mothers can be affected by the congenital heart block. Anti Ro/SS-A antibodies play a major pathogenic role in affecting the heart conduction tissue in this rare condition. PMID- 16218462 TI - Anti-SSA/Ro and anti-SSB/La antibody-mediated congenital heart block. AB - When anti-SSA/Ro antibodies are present in sera of mothers with connective tissue diseases, the incidence of congenital heart block (CHB) has been reported to be 1 2% in live births. The risk of recurrence of CHB in a subsequent child remains limited to 10-16%. CHBs are usually complete but CHB of the first or second degree can also be observed. In some cases, CHB is associated with endocardial fibroelastosis. Late-onset cardiomyopathy may occur later in life in these children. The mortality of CHB, which is predominant in utero and in the first months of life, is an estimated 16-19%. A pacemaker is required in about 66% of cases. Curative treatment of CHB is based on fluorinated steroids (dexamethasone or betamethasone) that do cross the placenta in an active form. Guidelines are available but further studies are needed to optimize treatment. PMID- 16218463 TI - Predictive and protective autoimmunity in cardiovascular diseases: is vaccination therapy a reality? AB - The role of the immune system in modulating atherosclerosis has recently been well documented. Studies have revealed that cellular and humoral immunity plays crucial roles in atherogenic plaque formation. This includes macrophages, CD4+ T cells and dendritic cells as well as autoantigens such as oxidized low density lipoprotein (oxLDL), heat shock proteins and beta2-glycoprotein I. Given these recent advances, various modifications of the immune system in experimental models have been proposed as therapeutic strategies, with the potential of inhibiting atherosclerosis progression. These modifications are switching the immune system (CD4+ T cells) from Th1 towards a Th2 anti-inflammatory cytokine secretion, and the induction of protective antibodies both of which may be induced by specific vaccination. Recent identification of specific immunoreactive antigenic epitopes on modified LDL, their successful implementation for immunization and the induction of atheroprotection, supports the idea that active vaccination may emerge as a novel immuno-modulating atheroprotective strategy. PMID- 16218464 TI - Idiopathic recurrent acute pericarditis: familial Mediterranean fever mutations and disease evolution in a large cohort of Caucasian patients. AB - Idiopathic recurrent acute pericarditis (IRAP) is suspected to be an autoimmune phenomenon. We studied 46 consecutive patients. We looked for: 1) the occurrence of new diagnoses of autoimmune diseases during our follow up; 2) HLA typing; and 3) the presence of the most frequent mutations linked to familial Mediterranean fever (FMF gene or MEFV). HLA typing was done in 21 patients at loci B, DRB1, DQA1 and DQB1. MEFV gene was looked in 23 patients using specific primers. During the follow-up we made a new diagnosis of primary Sjogren's syndrome in four patients (8.7%) and of rheumatoid arthritis in one patient (2.2%). HLA B14, DRB1*01 and DQB1*0202 were significantly more prevalent, but we did not find a typical HLA typing. MEFV gene was searched: exon 10 was checked by sequence and the E148Q mutation by restriction site analysis. No mutations were found. In conclusion, the prevalence of definite immunorheumatological diseases and the absence of the mutations linked to FMF reinforce the notion that idiopathic acute recurrent pericarditis is an autoimmune condition. PMID- 16218465 TI - Estrogens, autoimmunity and the heart. AB - Systemic inflammatory/autoimmune rheumatic diseases are associated with a significantly increased rate of atherosclerosis and cardiovascular disease. Several mechanisms of accelerated atherosclerosis have been proposed, including abnormal lipid and lipoprotein profiles, oxidative stress, enhanced apoptosis, thrombophilia, immune complexes and increased mononuclear cell infiltration of atherosclerotic lesions, local generation of cytokines and female estrogen deficiency. However, the widely shared enthusiasm about the cardioprotective potential of hormone replacement therapy (HRT) with estrogens, has come to an abrupt halt since very recent randomized trials failed to show a cardiovascular risk reduction in postmenopausal women. Several factors might play a role in these discrepancies, in particular, parts of the striking discrepancy between observational and randomized data have been attributed to an estrogen-mediated adverse effect on inflammation (enhancement, possibly dose-related). In fact, estrogens potentially increase the inflammatory/immune response in autoimmune rheumatic diseases. New roles for estrogen peripheral metabolites (hydroxylated) and their increased formation in inflammatory sites, might partially introduce some explanations for several apparently contrasting evidences. PMID- 16218466 TI - Cardiovascular involvement in rheumatoid arthritis. AB - Cardiovascular (CV) disease morbidity and mortality are increased in patients with rheumatoid arthritis (RA) and much of the excess CV disease morbidity appears to be due to atherosclerosis. The pathogenesis of atherosclerosis (ATS) in RA is complex and there is increasing evidence that many factors including novel and traditional cardiovascular risk factors, RA treatments and the RA inflammatory disease process are involved in the development of CV disease in these patients. Of particular interest are the effects of chronic inflammation and immune dysregulation associated with RA. These have been shown to be associated with endothelial dysfunction, which is an early, potentially reversible, functional abnormality of the arterial wall. However, as several CV disease risk factors and drug prescribing are also influenced by RA disease severity it is very difficult to separate out the effects of the inflammatory disease burden on the cardiovascular system in RA. PMID- 16218467 TI - Cardiac involvement in systemic lupus erythematosus. AB - Pericarditis is the most common cardiac abnormality in systemic lupus erythematosus (SLE) patients, but lesions of the valves, myocardium and coronary vessels may all occur. In the past, cardiac manifestations were severe and life threatening, often leading to death. Therefore, they were frequently found in post-mortem examinations. Nowadays cardiac manifestations are often mild and asymptomatic. However, they can be frequently recognized by echocardiography and other noninvasive tests. Echocardiography is a sensitive and specific technique in detecting cardiac abnormalities, particularly mild pericarditis, valvular lesions and myocardial dysfunction. Therefore, echocardiography should be performed periodically in SLE patients. Vascular occlusion, including coronary arteries, may develop due to vasculitis, premature atherosclerosis or antiphospholipid antibodies associated with SLE. Premature atherosclerosis is the most frequent cause of coronary artery disease (CAD) in SLE patients. Efforts should be made to control traditional risk factors as well as all other factors which could contribute to atherosclerotic plaque development. PMID- 16218468 TI - Atherogenesis and autoimmune disease: the model of lupus. AB - Accelerated atherosclerosis is a major cause of morbidity and mortality in patients with systemic lupus erythematosus (SLE). Certain 'classic' risk factors are associated with atherosclerosis risk in SLE. However, these factors alone do not fully explain the excess risk observed. Atherosclerosis is increasingly recognized as a chronic inflammatory condition and in SLE, complement activation and immune complex formation may promote atheroma development. Similarly, autoantibody production, especially those in the anticardiolipin (ACLA) family are gaining increasing attention. The role of steroids may not be completely straightforward, low doses may have a beneficial anti-inflammatory role whereas higher doses may exacerbate metabolic factors. In contrast, antimalarials have a beneficial effect on lipids as well as anti-inflammatory and anti-platelet effects. The aetiology of atherosclerosis in SLE is therefore multifactorial. A better understanding of the interface of autoimmunity and atherogenesis in the context of SLE will benefit lupus patients and will also help us better understand the pathogenesis of atherosclerosis in general. PMID- 16218469 TI - Cardiac involvement in the antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is a systemic autoimmune disease, associated with a hypercoagulable state and fetal loss and with other clinical manifestations including cardiac involvement. Cardiac manifestations of APS are valve abnormalities (valve thickening and vegetations), occlusive arterial disease (atherosclerosis and myocardial infarction), intracardiac emboli, ventricular dysfunction, and pulmonary hypertension. Antiphospholipid antibodies (aPLs) may have a role in the accelerated atherosclerotic arterial disease observed in APS, related to their ability to induce endothelial activation. aPLs have been incriminated in the pathogenesis of heart valve lesions in APS patients. Markers of endothelial cell activation are up-regulated with prominent deposition of aPL in heart valves, suggesting aPL deposition initiates an inflammatory process that recruits complement leading to the valve lesion. Autoantibody-mediated endothelial cell activation probably plays a role in sustaining a proadhesive, proinflammatory, and procoagulant phenotype. The heterogeneity of APS clinical manifestations is likely linked to the varied effects that aPL can induce on endothelial cells and to the different functions that endothelial cells display depending on the anatomic localization. PMID- 16218470 TI - From rheumatic fever to Libman-Sacks endocarditis: is there any possible pathogenetic link? AB - The heart lesions of rheumatic fever and the heart involvement in antiphospholipid syndrome (APS), have different clinical pictures. Yet, there are several common characteristics linking both diseases: 1) central nervous system (CNS) and heart involvement; 2) molecular mimicry between the a pathogen and the origin of the disease; 3) cross reacting antibodies between the pathogen and self molecules; 4) endothelial cell activation in the 'crime-area' i.e., the valves; 5) some of the patients with RF have circulating antiphospholipid antibodies, while APS may be associated with streptococcal infection; and 6) recently, a cross-reactivity between antibodies directed to the streptococcal M-protein and its synthetic derivative in rheumatic fever (RF) and antibodies derived from APS patients targeting the beta-2-glycoprotein-I (beta2GPI) and a beta2GPI related synthetic peptide. In the current paper, we summarize the possible links between the heart involvement in RF and APS. PMID- 16218471 TI - Heart involvement and systemic sclerosis. AB - Scleroderma heart involvement (SHI) is often manifest, and virtually always present when accurately searched and holds a significant prognostic value. Myocardial involvement by patchy fibrosis (secondary to both repeated ischaemia and immunoinflammatory damage) leads to ventricular diastolic dysfunction, whereas right ventricle overload and failure may complicate pulmonary hypertension. Left ventricular systolic dysfunction is present in a minority of patients, namely those presenting atherosclerotic coronary artery disease and/or arterial hypertension, sometimes triggered by sclerodermic renal involvement. Dysrhythmias and conduction disturbances are considered an hallmark of SHI, facilitated by autonomic dysfunction. SHI is frequently linked to parenchimal and/or vascular lung disease; they determine symptom occurrence, particularly dyspnoea, fatigue, palpitations and chest pain when pericardium is affected. Accurate cardiologic baseline screening and subsequent follow-up are mandatory in all patients, initially consisting in some noninvasive diagnostic procedures: visit, electrocardiogram (EKG), chest X-ray, Doppler-echocardiography. When needed, these examinations should be integrated by EKG Holter-monitoring, cardiopulmonary stress tests, cardiac magnetic resonance imaging, nuclear studies of myocardial function and perfusion, cardiac catheterization to better estimate pulmonary hypertension, and cardiac natriuretic hormone evaluation. Several vasodilator approaches (prostacycline or NO/endothelin) may counteract the microvascular dysfunction at peripheral and cardiopulmonary level, and fight the sequelae of pulmonary hypertension. PMID- 16218472 TI - Cardiac involvement in autoimmune myositis and mixed connective tissue disease. AB - The clinical cardiac manifestations most frequently reported in idiopathic inflammatory myopathies, myositis, are congestive heart failure, conduction abnormalities, that may lead to complete heart block and coronary artery disease. Although clinically overt cardiac involvement is rarely reported in myositis patients, subclinical manifestations are frequently observed and are predominated by conduction abnormalities and arrhythmias detected by ECG. Furthermore, cardiovascular manifestations constitute a major cause of death in myositis, thus cardiac involvement maybe overlooked in these patients. Also children with juvenile dermatomyositis may develop cardiac involvement although the frequency seems to be low. The underlying pathophysiologic mechanisms that may cause cardiac manifestations could involve myocarditis and coronary artery disease as well as involvement of the small vessels of the myocardium. In patients with mixed connective tissue disease (MCTD) clinically significant cardiac involvement is also rare, the most frequently reported manifestations being pericarditis and pulmonary hypertension, the latter often attributable to small vessel disease, and often a prognostic unfavourable manifestation. PMID- 16218473 TI - Pulmonary arterial hypertension associated to connective tissue diseases. AB - Pulmonary arterial hypertension is a well-known complication of connective tissue diseases such as systemic sclerosis, systemic lupus erythematosus, mixed connective tissue diseases, and to a lesser extent, rheumatoid arthritis, dermatopolymyositis and primary Sjogren's syndrome. In these patients, pulmonary hypertension may occur in association with left heart disease, interstitial fibrosis or as a result of a isolated pulmonary arteriopathy. The incidence of pulmonary arterial hypertension in the limited form of systemic sclerosis is about 10%. The pathophysiologic mechanisms leading to pulmonary arterial hypertension remain unknown. Symptoms and clinical presentation are very similar to idiopathic pulmonary arterial hypertension but mortality was confirmed to be higher. Echocardiography is the reference investigation for the detection of pulmonary arterial hypertension but the results should be confirmed by right heart catheterization. Treatment appears more complex as compared to idiopathic pulmonary arterial hypertension. Intravenous epoprostenol therapy has been shown to be effective in a special trail. Also, the endothelin receptor antagonists bosentan and sitaxentan, the phosphodyesterase-type-5 sildenafil and subcutaneous treprostinil have shown favourable results. PMID- 16218474 TI - Cardiac involvement in small and medium-sized vessel vasculitides. AB - The heart can be involved in vasculitides but the frequency of its involvement and the manifestations vary according to the vasculitis. Cardiovascular manifestations include cardiomyopathy (specific or resulting from myocardial infarctions), coronary arteritis (with risk of aneurysms, stenoses and thrombosis formation or rupture), pericarditis, valvulitis, conduction-tissue involvement (with heart blocks), arrhythmias (mainly supraventricular) and/or dissection of the aorta (and/or its proximal branches). As many of these manifestations are clinically silent, at least during their early stages, heart function should be systematically assessed in vasculitis patients, with at least ECG and echocardiography, and more invasive exploratory procedures when the former reveal abnormalities or symptoms become manifest. Specific cardiomyopathy has been identified as a factor of poor outcome in small and medium-sized vessel vasculitides (five-factor score). Therefore, in addition to symptomatic treatments, prescription of corticosteroids and immunosuppressants (mainly cyclophosphamide) is considered mandatory. This regimen has dramatically improved the overall prognosis of affected patients. PMID- 16218475 TI - Behcet's disease and cardiovascular involvement. AB - Behcet's disease (BD) is a multisystem disease of unknown etiology characterized by chronic relapsing orogenital ulcers, uveitis and systemic involvement including articular, gastrointestinal, cardiopulmonary, neurological and vascular pathology. The incidence and nature of cardiac involvement are not clearly elucidated. Cardiovascular manifestations have been reported in 7-46% of patients and mortality occurs in up to 20% of those patients with marked vascular involvement. Sporadic cases of endocarditis, myocarditis, pericarditis, acute myocardial infarction, aortic aneurysm, ventricular thrombosis, congestive cardiomyopathy and valvular dysfunction have been reported. This review discusses the general aspects of the pathogenic mechanisms and clinical features cardiovascular involvement in BD, and provides the data of cardiovascular involvement in a cohort of Italian BD patients. PMID- 16218476 TI - Cardiac imaging techniques in systemic autoimmune diseases. AB - Systemic autoimmune disorders are frequently associated to cardiac involvement and to a high prevalence of ischemic coronary events, often occurring at a younger age than in the normal population. Large increase in mortality is related to premature atherosclerosis with coronary artery disease and stroke in patients with connective tissue diseases. Coronary heart disease is responsible for 40-50% of the deaths of patients with rheumatoid arthritis. Transesophageal or transthoracic echocardiography are the most useful and noninvasive techniques able to detect not only valvular abnormalities, embolic sources or pulmonary hypertension, but also left ventricular systolic or diastolic dysfunction. Furthermore, the introduction of new indexes, contrast agents and software increased the accuracy of this technique. It is possible now to evaluate coronary flow reserve by transthoracic echocardiography in patients with systemic autoimmune disease in order to detect microvasculature disorder. However, an ischemic response in a symptomatic patient requires, in most cases, further evaluation with cardiac catheterization. Coronary artery imaging allows confirmation of the presence, extent and position of atheromatous lesions. More recently, other imaging modalities including magnetic resonance and computerized tomography angiography have been developed to allow imaging of the coronary arteries. PMID- 16218477 TI - T cells and cytokines in atherogenesis. AB - Recent findings suggest that inflammation plays a key role in atherosclerosis from the earliest stage of lesion initiation, to the ultimate complication of thrombosis. In patients who died because of acute coronary syndromes (ACS), coronary atherosclerotic plaques are characterized by the presence of macrophages, and to a lesser extent T-lymphocytes, at the immediate site of either plaque rupture or superficial erosion. The rupture-related inflammatory cells are activated, indicating ongoing inflammation. ACS patients are also characterized by activated circulating lymphocytes, monocytes and neutrophils, and by increased concentrations of proinflammatory cytokines and of the highly sensitive acute phase reactant C-reactive protein. Interestingly, an unusual subset of T cells, CD4+ CD28null T cells, involved in vascular complication of rheumatoid arthritis because of their functional profile predisposing for vascular injury, are expanded in the peripheral blood and infiltrate the coronary lesions of ACS patients. The presence of activated T lymphocytes implies antigenic stimulation, but the nature of such antigen(s) remains to be investigated. Several autoantigens expressed in the atherosclerotic plaque, including oxidized LDL and heat shock proteins, and infectious agents are able to elicit an immune response. The inflammatory component is not localized to the 'culprit' plaque, but it is diffused to the entire coronary vascular bed, and involves also the myocardium. PMID- 16218478 TI - Oxidized LDL/beta2-glycoprotein I complexes: new aspects in atherosclerosis. AB - beta2-glycoprotein I (beta2GPI) is a major antigenic target for antiphospholipid antibodies. Oxidized low-density lipoprotein (oxLDL) is the principal lipoprotein found in atherosclerotic lesions, and it colocalizes with beta2GPI and immunoreactive lymphocytes. oxLDL/beta2GPI complexes appeared in the blood circulation of patients with diseases, such as systemic lupus erythematosus (SLE), antiphospholipid syndrome (APS), systemic sclerosis, diabetes mellitus and chronic renal diseases. Thus, the complexes may be associated with systemic and chronic inflammation of the vasculature. IgG anti-oxLDL/beta2GPI complexes autoantibodies and their immune complexes were detected only in SLE/APS patients and in its animal model and were strongly associated with arterial thrombosis. The oxLDL/beta2GPI complexes were internalized by macrophages via IgG anti beta2GPI antibody-mediated phagocytosis. In contrast, IgM anti-oxLDL antibodies derived from hyperlipidemic mice reduced the incidence of atherosclerosis. The distribution patterns of IgG and IgM anti-oxLDL antibodies in patients suggest the different roles of these antibodies. PMID- 16218479 TI - Role of antiheat shock protein 60 autoantibodies in atherosclerosis. AB - Immuno-inflammatory processes are implicated, as one of the prime pathogenic processes involved, in the development and progression of early atherosclerosis. High levels of circulating antiheat shock protein 60 (HSP60) autoantibodies have been associated with increasing severity of atherosclerosis in patients. We have recently presented evidence, extending this statistical association to that of causality, by showing that anti-HSP60 antibodies purified from sera of patients with documented atherosclerosis when injected into tail vein of apoE deficient mice resulted in accelerated atherosclerosis in them. High degree of sequence homology between microbial and mammalian HSP60, due to evolutionary conservation, carries a risk of misdirected autoimmunity against HSPs expressed on the stressed cells of vascular endothelium. HSPs and anti-HSP antibodies have been shown to elicit production of pro-inflammatory cytokines. These autoimmune reactions to HSPs expressed in the vascular tissue can contribute to both initiation and perpetuation of atherosclerosis. PMID- 16218480 TI - Innate immunity and atherogenesis. AB - Atherosclerosis is recognized as the pathological basis of cardiovascular disease (CVD) and recent advances in basic science have shown that it should be considered as a chronic inflammatory process. Both elements of the innate and the adaptive immunity appear to be actively involved in atherogenesis. In fact, the potential role played by pattern-recognition receptors (Toll-like receptors and scavenger receptors), cytokines (such as IL-1, IL-6, TNFalpha), chemokines and pentraxines (such as CRP and PTX3) represents an emerging field of investigation in atherogenesis. In the near future we expect a better definition of the real biological and clinical impact on CVD of these mediators. On one side, they could become useful to complement traditional risk factors, in order to identify new categories of subjects prone to CVD development. On the other, they could become an additional potential target for therapeutic strategies. PMID- 16218481 TI - CRP is or is not a reliable marker of ischaemic heart disease? AB - In recent years a growing body of evidence has emphasized the role of C-reactive protein (CRP) as a marker of future cardiovascular events. CRP is a pentameric molecule widely utilized as a marker of infections and inflammation. The evidence that inflammation plays an important role in the pathogenesis of coronary artery disease and in plaque destabilization has lead to use of CRP as a marker of cardiovascular disease as well. First described as a component of the inflammatory pathway in acute coronary syndromes, CRP has been consistently found to be associated with the risk of future events in no-ST elevation acute coronary syndromes, independently of other risk factors, including troponine. Subsequently CRP has been described as a powerful marker of risk of future events in large populations of apparently healthy subjects. So far there is very little doubt that CRP represents a reliable marker of cardiovascular events, but some issues remain unanswered such as why CRP is a good marker of cardiovascular events and whether or not a better inflammatory marker exists. It must be stressed that CRP, because of its analytical and biological properties and the large amount of available data, is the only inflammatory marker accepted for clinical use. PMID- 16218482 TI - COX-2 and prostaglandins in atherosclerosis. AB - Arachidonic acid metabolism is involved in acute ischemic syndromes affecting the coronary or cerebrovascular territory, as demonstrated by biochemical measurements of eicosanoid biosynthesis and the results of inhibitor trials in these settings. In particular, the efficacy of low-dose aspirin in reducing the complications of acute ischemic syndromes has focused attention on the cyclooxygenase (COX) pathway of arachidonic acid metabolism and its products, collectively termed prostanoids. Two cyclooxygenase (COX)-isozymes have been characterized, COX-1 and COX-2, that differ in terms of regulatory mechanisms of expression, tissue distribution, substrate specificity and preferential coupling to upstream and downstream enzymes. While the role of platelet COX-1 in acute ischemic diseases is firmly established, the role of COX-2 in atherothrombosis is still uncertain. Studies from our group suggest that variable expression of upstream and downstream enzymes in the prostanoid biosynthesis may represent important determinants of the functional consequences of COX-2 expression and inhibition in different clinical settings. PMID- 16218483 TI - Oxidant stress, inflammation and atherogenesis. AB - Low-grade inflammation, enhanced oxidant stress and lipid peroxidation have been shown in association with increased cardiovascular risk associated with cardiovascular events. It has been hypothesized that the low-grade inflammatory state characterizing metabolic disorders such as obesity, hypercholesterolemia, type 2 diabetes mellitus and homozygous homocystinuria may be the primary trigger of thromboxane-dependent platelet activation mediated, at least in part, through enhanced lipid peroxidation. Interestingly, as the clinical course of systemic lupus erythematosus (SLE), in particular in the presence of antiphospholipid antibodies, may be complicated by vascular disease, several mechanisms contributing to vascular complications have been documented also in this setting, including enhanced lipid peroxidation and thromboxane biosynthesis. Although epidemiological studies show an inverse relationship between antioxidant vitamin intake and cardiovascular disease, several clinical trials have obtained conflicting results on the effects of vitamin E on the risk of cardiovascular events. The availability of analytical tools for measuring F2-isoprostane biosynthesis in man has improved our understanding of the interplay between lipid peroxidation and low-grade inflammation. The use of F2-isoprostane as a biochemical end-point for dose-finding studies may allow reassessing the adequacy of vitamin supplementation in different clinical settings. PMID- 16218484 TI - Statins and autoimmune diseases. AB - Besides the well-known lipid-lowering effect, statins display nonlipid-lowering pharmacological activities. In vitro and in vivo studies suggest that statins have direct anti-inflammatory, anti-thrombotic and plaque-stabilizing effects via a number of mechanisms. A direct immunomodulatory effect has been also demonstrated in in vitro and in vivo experimental models. In addition to traditional risk factors, systemic inflammation, immune-mediated responses and thrombophilia have been suggested to play a major role in sustaining the premature atherosclerosis in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus. This review focuses on the anti-inflammatory and immunomodulating mechanisms of statins as demonstrated in in vitro and in vivo experimental models, providing new insights for the use of statins in treating systemic autoimmune diseases both for their anti-atherosclerotic activity and for their pleiotropic effects on inflammation, haemostasis and the immune responses. PMID- 16218485 TI - Pharmacological modulation of plaque instability. AB - Atherosclerotic plaque rupture is promoted by metalloproteinase (MMP)-2 and MMP 9, enzymes that degrade the fibrous cap leading to plaque erosion. MMP biosynthesis is mediated by prostaglandin (PG)E2, the product of cyclooxygenase (COX)-2/inducible PGE synthase (mPGES) activity. We have recently reported the overexpression of COX-2/mPGES-1 in vulnerable plaques as a basis of MMP-mediated plaque instability. Hypercholesterolemia and hypertension are two important risk factors for atherosclerosis. Recent trial showed that statins and AT1 receptor blockers significantly reduce the incidence of cardiovascular events in humans. Since anti-inflammatory effects have been reported in association to therapy with statins or AT1 receptor blockers, in two different studies we hypothesized that these drugs can stabilize atherosclerotic plaques through modulation of COX 2/mPGES-1-dependent MMP biosynthesis. Our data demonstrated the stabilizing effect of atherosclerotic plaques by simvastatin or irbesartan, that is due, at least in part, to the reduction of inflammatory burden and suppression of PGE2 dependent metalloproteinases release. PMID- 16218486 TI - Bisphosphonates and atherosclerosis: why? AB - The increasing knowledge on bone calcification processes has revealed some similarities with vascular tissue, where calcifications of arteries and cardiac valves contribute to several cardiovascular problems, such as heart failure, systolic hypertension, and myocardial and peripheral ischemic disease. Bisphosphonates have been used extensively for over two decades for the treatment of diseases associated with excessive bone resorption, i.e., osteoporosis, osteolytic bone metastasis, hypercalcemia and Paget's disease, by blocking osteoclastic function. Etidronate, pamidronate and clodronate has been shown to inhibit the development of experimental atherosclerosis, and proposed mechanisms for this action include inhibition of arterial calcification and lipid accumulation, degradation of atherogenic LDL-cholesterol and reduced foam cell formation. Bisphosphonates inhibit various enzymes involved in cholesterol biosynthesis and suppress macrophages in atheromatous lesions. The possibility of pharmacological agents that effectively treat both osteoporosis and atherosclerosis is attractive, however, current evidence is not conclusive and further research is necessary to confirm these actions in the clinical setting. PMID- 16218488 TI - Cardiovascular effects of coxibs. AB - A new class of drugs, the selective cyclooxygenase-2 (COX-2) inhibitors, or coxibs, have recently been marketed as an alternative to conventional nonsteroidal anti-inflammatory drugs (NSAIDs) on the basis of a lower risk of gastrointestinal side effects. The recent withdrawal of rofecoxib, along with safety concerns about other COX-2 selective inhibitors raises important questions about the cardiovascular toxicity of these drugs. Recently some concerns arose even for a possible cardiotoxicity of nonselective nonsteroidal anti-inflammatory drugs. From data available so far, it seems that coxibs still remain a rational choice for patients with low cardiovascular risk and high gastrointestinal risk. Long-term studies with a cardiovascular endpoint involving both selective and nonselective anti-inflammatory drugs are warranted. PMID- 16218487 TI - Tumor necrosis factor-alpha, biologic agents and cardiovascular risk. AB - The increased risk of premature cardiovascular disease (CVD) in rheumatoid arthritis (RA) patients may depend on traditional risk factors but may also be attributable to RA-specific risk factors such as disease-related dyslipidemia, or cytokines such as tumor necrosis factor-alpha (TNF-alpha). TNF-alpha is a proinflammatory cytokine that can produce widespread deleterious effects when expressed in large amounts. It is produced in the heart by both cardiac myocytes and resident macrophages under conditions of cardiac stress, and is thought to be responsible for many of the untoward manifestations of cardiac disease. TNF-alpha may play a role in the triggering and perpetuation of atherosclerosis. Treatment with biologic agents directed against TNF-alpha has significant clinical benefits in inflammatory diseases such as RA and may be able to reduce cardiovascular risk. The disappointing results of the recent studies to antagonize TNF-alpha in CVD may have various explanations. However, the effects of TNF-alpha blockers on incident cases of congestive heart failure (CHF) in RA remains controversial. Due to the lack of evidence of a beneficial effect of anti-TNF-alpha agents in treatment of CHF, they should not be used to treat patients with New York Heart Association (NYHA) class III or IV heart failure. PMID- 16218490 TI - Thiazolidinediones and inflammation. AB - Thiazolidinediones (TZDs) are selective ligands of peroxisome-proliferator activated receptor gamma increasingly used in the treatment of type 2 diabetes. Both in vitro and in vivo studies provide evidence that TZDs have anti inflammatory properties. TZDs inhibit macrophage activation and decrease inflammatory cytokine expression and release in macrophage and monocyte. In vivo, treatment with TZDs decreases circulating mononuclear cells nuclear NF-kB content while increasing, in the same cells, expression of IkB, an NK-kB inhibitor. Furthermore, TZD treatment results in decreased plasma levels of inflammation and cardiovascular risk markers such as CRP, MMP9, PAI-1 and sCD40 in both obese and type 2 diabetic patients. Finally, TZDs induce synoviocyte apoptosis and reduce secretion of TNFalpha, IL-6 and IL-8 in synoviocyte from rheumatoid arthritis patients. TZDs might thus be considered for use in clinical trials targeting prevention of atherosclerosis and cardiovascular diseases and in pilot trials exploring the possibility that TZDs might help in the treatment of rheumatic diseases. PMID- 16218489 TI - New trends in anticoagulant treatments. AB - Classic anticoagulant drugs are very effective, save lives and have been used for more than 50 years. Nevertheless, some drawbacks are encountered in their routine clinical use. Recently, pharmaceutical research has developed new drugs, some of which are already on the market. This is the case of fondaparinux, a pentasaccharide which can interact with antithrombin, thus inhibiting factor Xa. Modification of its structure (idraparinux) has led to more stable binding with antithrombin and to an increase in its half-life allowing for once-a-week administration. Another important oral compound is ximelagatran which directly binds thrombin and blocks its catalytic site. There is no need for laboratory control, and phase II and phase III studies are encouraging. Thus, in the next few years, we may witness great changes in the treatment of patients with thromboembolic disorders. PMID- 16218491 TI - [Management of behavioral risk factors for cardiovascular disease (CVD)]. AB - Changes in many patterns of individual risk behaviors (unhealthy diet, smoking, sedentary lifestyle) are necessary in a large majority of patients with established CVD or at high risk of CVD. Make healthy food choices. Energy intake must be adjusted to maintain ideal body weight. Eating from each major food group will ensure dietary balance. The consumption of fruits and vegetables, whole grain cereals and bread, low fat dairy products, fish and lean meat should be encouraged. Oily fish and omega-3-fatty acids are specially recommended. Saturated and trans fatty acids should be replaced with MUFA's or PUFA's of vegetable and seafood origins. Totalfat intake should account for no more than 30% of energy intake, and intake of saturated fats should not exceed a third of total fat intake. The intake of cholesterol should be less than 300 mg/day. Stop smoking tobacco. All smokers should be professionally encouraged to permanently stop smoking all forms of tobacco. Increase physical activity. A lack of regular physical activity may contribute to the early onset and progression of Cardiovascular Disease. Regular physical activity and the maintenance of physical fitness is beneficial in all age groups and should be promoted as an integral part of cardiovascular prevention. Healthy people should be advised to choose enjoyable activities, which fit into their daily routine, preferably 30-40 min, 4 5 times weekly at Borg 13 or at 60-75% of the average maximum heart rate. For patients with established CVD, advice must be based on a comprehensive clinical judgement including the results of an exercise test. PMID- 16218493 TI - [Stress as a cardiovascular risk factor]. AB - Stress is a reaction of the organism to a stressor with various negative health consequences including the development of arteriosclerosis. Stress may cause myocardial ischemia, rhythm disturbances, platelet stimulation, increased blood viscosity through hemoconcentration as well as endothelial dysfunction and coronary vasoconstriction in the presence of arteriosclerosis of the coronary arteries. Psychosocial risk factors may also lead to the development and to clinical consequences of arteriosclerosis. The five most important psychosocial risk factors are: depression, anxiety, personality factors, social isolation and chronic life stress. Various components are responsible for the fact that life events or certain psychosocial factors can lead to a stress reaction. Psychosocial risk factors have direct pathophysiologic effects but also influence life style and behavior Various stress management intervention programs have proven to be effective. Psychosocial intervention in conjunction with cardiac rehabilitation programs has shown to reduce cardiovascular morbidity and mortality. PMID- 16218492 TI - [Socioeconomic class as a risk factor for cardiovascular diseases]. AB - It's been known for a long time, that certain diseases are more frequent in lower socioeconomic classes. But knowledge about the nature of this association, its main risk factors and how to improve health outcomes in lower social groups is still limited. Social class has been defined by different indicators by e.g. occupation and job position or the highest school qualification achieved. For international comparisons different classifications such as "The Registrar General's Social Class Classification " or the "International Standard Classification of Education" have been used. Several European Studies show a higher prevalence of cardiovascular diseases and cardiovascular risk factors including smoking, obesity, physical inactivity, hypertension and hypercholesterolemia in lower socioeconomic classes. But this studies also show that all socioeconomic groups have access to medical services. The Data from the Swiss Health Survey show the distribution of cardiovascular risk factors and diseases by three levels of education: Behaviouralfactors such as smoking, obesity and physical inactivity are more commonly present in the lower socioeconomic groups. People with a lower educational level visit their GP more often, whereas people with a higher level of educational consult specialists more frequently. Medical services are often used to check of blood pressure, blood sugar and cholesterol. An indication of state of health may be shown by medication and treatment for cardiovascular disease which is more prevalent in lower socioeconomic groups. The present discussion of explanations of the poorer state of health in lower socioeconomic groups goes beyond the classical risk factors for cardiovascular disease. Studies have shown that after the correction for risk factors a correlation remains between social class and state of health. It is believed, that psychosocial factors such as self-esteem, control in the workplace or coping-strategies play an additional important role in staying healthy. Therefore prevention of cardiovascular disease should not be restricted to minimisation of risk factors or to the measurement of certain blood-values and treatments. Efforts must also go into creating the conditions for a healthier life and promote possibilities for healthier behaviour in all socioeconomic classes. PMID- 16218494 TI - [Salt, hypertension and cardiovascular risk]. AB - For more than a century salt and blood pressure have been linked. Prospective randomized clinical trials and meta-analyses provided evidence that a reduction of daily salt intake of 100 mmol will lower systolic blood pressure by 3-5 mm Hg, whereas diastolic blood pressure can be reduced by 1 mm Hg. The effect of salt restriction depends strongly on individual salt sensitivity. Whether a reduction of salt intake will ultimately result in a decline of cardiovascular morbidity and mortality remains to be determined in future studies. PMID- 16218495 TI - [Fruit and vegetable]. AB - An increased fruit and vegetable intake is associated with a reduced risk for chronic diseases. The protective effects are due to different mechanisms such as an increased intake of essential micronutrients as well as different non nutritive phytochemicals. A dietary pattern characterised by a high intake of fruits and vegetables has a much higher protective effect than the intake of pharmacological doses of single nutrients or phytochemicals. PMID- 16218496 TI - [Egg intake and cardiovascular risk]. AB - Egg yolk has the highest content of cholesterol observed in different types of food. Intake of one egg per day increases the serum concentration of LDL cholesterol by 0.10 mmol/l. In healthy people eating a Western diet, egg consumption does not correlate with cardiovascular risk. However, a significant increase of the risk was observed in diabetic subjects starting from an intake of 5-7 eggs per week. However, dietary pattern rather than a single component of nutrition influences cardiovascular risk. The egg - rich in proteins, unsaturated fatty acids, vitamins and minerals - should be part of our nutrition, and it is not justified to recommend a general limitation of egg intake. However, we do not advice unbalanced high egg consumption. A cardioprotective diet is characterized by high variability and contains plenty of fruits, vegetables, and complex carbohydrates. PMID- 16218497 TI - [Antioxidant vitamin supplementation in the prevention of cardiovascular disease]. AB - Oxidative stress, in particular oxidative modification of LDL-cholesterol, appears to be of great importance in the pathogenesis of atherosclerosis. Various observational epidemiological studies have suggested that antioxidant vitamin intake is associated with reduced cardiovascular morbidity and mortality. Also, experimental studies in animals have demonstrated that antioxidant vitamins slow the progression of atherosclerosis. However, prospective controlled clinical trials have failed to demonstrate a benefit of antioxidant vitamin supplementation in primary or secondary prevention of cardiovascular disease. Thus, the use of antioxidants and vitamin supplements as a preventive or therapeutic intervention can not be recommended. PMID- 16218498 TI - [Alpine cheese--healthy for your heart or just a lifestyle product?]. AB - Until the 19th and the early 20th century, milk and milk products, particularly of alpine origins, seemed of special nutritional and health value and were highly recommended for the therapy resp. the therapeutic adjunct of various diseases, particularly for pulmonary tuberculosis. More recently, the association of saturated fat intake and arteriosclerosis led to the reduced use of milk and cheese resp. to the introduction of low-fat milk products. Again, alpine milk and cheese seem to differ somewhat from the others, since they appear to contain 4 times more alpha linolenic acid, three times more conjugated linoleic acid, a lower n-6:n-3 ratio, more total n-3 fatty acids and less palmitic acid as a measure of total saturated fat compared to cheese produced with silage feed in the lowlands (e.g. english cheddar). Even cheese from cows fed with linseed supplementations did not reach the n-3 concentrations of the alpine probes. Thus, alpine milk products from cows kept traditionally, and fed predominantly with alpine grass seem to have an interesting cardiovascular and possibly an economically favourable potential. PMID- 16218500 TI - [Coffee--poison or medicine?]. AB - Intake of coffee, one of the most common beverages worldwide, has often been discussed as a potential cardiovascular risk factor. However, definitive data about this topic are missing and newer studies even point out for a favorable rather than hazardous effect. Despite many studies no clear association between coffee and the risk of hypertension, myocardial infarction and other cardiovascular diseases was found. Recent publications suggest that moderate coffee intake does not represent a health hazard, but may even be associated with beneficial effects on the cardiovascular system and diabetes. PMID- 16218499 TI - [Fats and oils (including omega3, omega6)]. AB - Saturated fatty acids contribute essentially to atherogenesis, especially to coronary artery disease. In contrast, the protective effect of monounsaturated fatty acids such as oleic acid, contained in olive oil and as a constituent of a Mediterranean diet is very well shown epidemiologically. There is a modest beneficial effect on the clinical manifestation of coronary heart disease when saturated fatty acids of animal provenience are partially replaced by omega 6 fatty acids (linoleic acid). However, studies with the addition of omega-3-fatty acids in the diet (fish oil rich in eicosapentaenoic and docosaexaenoic acid and rape seed oil rich in alpha linolenic acid) demonstrate a significant decrease of sudden cardiac death and non fatal myocardial infarction. Long-chain omega-3 fatty acids have a direct antiarrhythmic effect on myocytes. The reduction of non fatal myocardial infarctions during consumption of diets rich in long chain omega 3-fatty acids could at least in part be attributed to inhibitory effects on platelet aggregation and thus on thrombus formation and to a stabilization of atherosclerotic plaques. PMID- 16218501 TI - [The sweet secret of dark chocolate]. AB - For centuries dark chocolate has been known for its taste as well as its beneficial effects on health. Mainly polyphenols, a heterogeneous group of molecules, have been associated with antioxidant and immunomodulatory properties. Furthermore they inhibit primary hemostasis and pathways associated with platelet activation and aggregation. PMID- 16218502 TI - [Alcohol and risk of coronary heart disease]. AB - More than 60 prospective cohort studies have shown a consistent association between regular and moderate alcohol consumption and decrease in risk of coronary heart disease, ischemic stroke and heart failure by 20 to 40% as compared to heavy alcohol intake or drinking no alcohol. Lower protective effects were found in young, in women and in men living outside the Mediterranean area. Moreover, some biological characteristics of alcohol, particularly red wine, could interfere with the athero-thrombotic process and contribute to increase the plausibility for the protective effects of alcohol on cardiovascular diseases. However, the results of meta-analyses also demonstrate harmful effects in relation with dose and pattern of alcohol consumption. In regard to the available scientific data, alcohol consumption cannot be include in the recommendations for the prevention of cardiovascular diseases. On the other hand alcohol should not be prohibited when consumption remains mild to moderate. PMID- 16218503 TI - [Homocystein--an independent risk factor for cardiovascular and thrombotic diseases]. AB - Over the last 20 years homocysteine has taken on increasing importance as an independent, potentially modifiable risk factor for various forms of vascular disease including peripheral and cerebral vascular disease, coronary heart disease and thrombosis. This association has been ascertained in many retrospective and prospective studies but the strength of risk is not yet firmly established although it is clearly dependent on several modifying factors such as other risk factors, nutrition and genetic polymorphisms. Generally it is estimated that hyperhomocysteinaemia is responsible for about 10% of all risks. Homocysteine is formed from the dietary amino acid methionine and plays a pivotal role in folate metabolism and methyl group transfer. Its concentrations in tissues and plasma are influenced by many genetic and environmental factors, especially vitamins such as folate, B12 and B6 as well as certain medications and even life style factors. Nowadays the measurement of plasma homocysteine is freely available although care has to be taken in sample handling and interpretation of results. Final proof that homocysteine is a causal agent and not just a marker for cardiovascular disease and that reduction of plasma homocysteine by vitamin treatment reduces risk of cardiovascular disease is still awaited. Therefore at the present time neither wide-scale screening for homocysteine levels nor general prophylaxis with high dose vitamins is justified. However most experts recommend homocysteine determination in individuals with existing or high risk for arterial or venous blood vessel disease and their relatives. Elevated homocysteine can be lowered in such cases with a combination of folic acid, vitamin B12 vitamin B6. The results of ongoing trials on the impact of such treatment on risk of vascular disease are awaited with great interest. PMID- 16218504 TI - [Low-carbohydrate diets for obesity]. AB - Low-carbohydrate diets for the treatment of obesity are currently receiving widespread popularity, mainly in Anglo-Saxan countries. Several controlled trials have demonstrated larger weight loss (3-6 kg) during these diets compared to conventional low-fat diets. The weight differences between these diets were statistically significant after 6 months but not any more after 12 months. Possible reasons for the increased weight loss may be the initial loss of body water, later on the diminishment of food choices, the satiating properties of proteins and the anorectic effect of ketosis. Controlled studies have not demonstrated any increase in LDL-cholesterol but a favorable increase in HDL-C and a lowering of serum triglycerides. Even if these diets "work" in selected patients, they are problematic because the increase in protein ingestion may lead to increased intake of saturated fat, and intake of water soluble antioxidants, vitamins and of fiber may be too low. For these reasons, their protective effect on the "major killers " atherosclerotic diseases, hypertension, diabetes type 2 and some forms of cancer is questionable. Moreover, these diets are unpleasant on the long run, and social eating is impaired. For all these reasons they should not be generally recommended. Reasonable nutritional changes for the treatment of obesity should be suitable for lifelong use, and they should always be combined with increased physical activity and with behavioral measures. PMID- 16218505 TI - [Sedentarity--sedentary lifestyle and physical activity]. AB - Physical inactivity is the most important planetary reason for non-transmissible mortality. Technical developments have allowed a sedentary lifestyle. This causes health problems such as insulin resistance, atherosclerosis, heart failure and obesity. In addition, disturbances of bones and muscles as well as dementia of the Alzheimer type are associated with sedentarity. Assessing this risk factor and attempting to increase physical activity should be a very important part of any general practitioner's measures. PMID- 16218506 TI - [Smoking cessation--a central role for physicians]. AB - Physicians are in a unique position to advice smokers to quit by the ability to integrate the various aspects of nicotine dependence. This review provides an overview of intervention with smokers presenting in a primary care setting. Strategies used for smoking cessation counseling differed according to patient's readiness to quit. For smokers who do not intend to quit smoking, physicians should inform and sensitize patients about tobacco use and cessation, especially by personalizing benefits to quit and challenging smokers 'beliefs. For smokers who are dissonant, physicians should use motivational strategies, such as discussing barriers to cessation and their solutions. For smokers ready to quit, the physician should show strong support, help set a quit date, prescribe pharmaceutical therapies for nicotine dependence, such as replacement therapy and/or bupropion, with instructions for use, and suggest behavioral strategies to prevent relapse. PMID- 16218507 TI - Cause or effect of increased inflammation? PMID- 16218508 TI - An update on the impact of Staphylococcus aureus enterotoxins in chronic sinusitis with nasal polyposis. AB - Nasal polyps in adults, characterized by abundant eosinophils, local overproduction of immunoglobulin E, and often associated with asthma, have been appreciated as an eosinophilic inflammation, potentially of allergic origin, but unrelated to a bacterial impact. Evidence accumulates, however, that Staphylococcus aureus colonizes chronic rhinosinusitis with, but not without polyps, with significantly increased prevalence. The germs release enterotoxins, which act as superantigens and induce a topical multiclonal IgE-formation as well as a severe, possibly steroid-insensitive eosinophilic inflammation. Recently, S. aureus could be demonstrated to reside intraepithelially, and potentially to release superantigens into the tissue from within the epithelial cells. An immune defect, either in the innate or adaptive immunity, might be responsible for this phenomenon. Follicle-like structures and lymphocyte accumulations, specifically binding enterotoxins, can be found within the polyp tissues, giving rise to local IgE formation. The superantigen-induced immune response also leads to a modulation of the severity of the eosinophilic inflammation, and may be linked to lower airway co-morbidity in polyp patients. Interestingly, IgE antibodies to enterotoxins can be found in the majority of aspirin-sensitive polyp tissues, associated with a substantial increase in ECP and IL-5. The possible role of S. aureus enterotoxins in polyp disease in Europe, the US and Asia has meanwhile been supported by several studies, demonstrating the presence of IgE antibodies to enterotoxins and inflammatory consequences in nasal polyp tissue. First studies also point to an involvement of S. aureus derived enterotoxins in lower airway disease, such as severe asthma and exacerbated COPD, clearly suggesting a clinical need for diagnosis and treatment of the germ and its related effects. Therapeutic approaches are so far empirical, and need further study, also serving to proof the clinical relevance of the concept. PMID- 16218509 TI - Consensus report on acoustic rhinometry and rhinomanometry. AB - With respect to acoustic rhinometry, new elements concern the problems related to the interpretation of the minimal cross-sectional area, and the presentation of a protocol for a multinational study, which aims to define a normal nose. Also, the previously issued recommendations for standardisation for technical specifications and standard operating procedures are briefly reviewed. For rhinomanometry, new insights into the field of fluid dynamics are highlighted, as well as their repercussion on more recent graphical representations for active anterior rhinomanometry such as four phases rhinomanometry and resistometry. For acoustic rhinometry as well as rhinomanometry, a more stringent standardisation of decongestive procedures is suggested. PMID- 16218510 TI - Site of origin of nasal polyps: relevance to pathogenesis and management. AB - The site of origin of sino-nasal polyps was documented in 113 consecutive patients undergoing functional endoscopic sinus surgery (FESS). These patients were assigned pre-operatively to 4 clinical groups according to the out-patient recorded endoscopic appearance of their nasal cavities; chronic rhinosinusitis without polyps (CRSS) n=35, grade 1 polyps n=28, grade 2 polyps n=30 and grade 3 polyps n=20. In the group of patients diagnosed with polyps pre-operatively, 97.4% had polyps originating in the anterior ethmoid complex, of which 89.7% had polyps originating in the anterior ethmoidal cells and over 60% had polyps specifically originating from each of the following sites: the uncinate or infundibulum, the posterior ethmoid sinus, the frontal recess and the face of the bulla ethmoidalis. In the group diagnosed pre-operatively as CRSS without polyps, polyps were found in 60% of patients within the sinuses during surgery. In summary, our findings suggest that polyps originate from the middle meatus, and may be found at surgery when undetectable at pre-operative endoscopy. PMID- 16218511 TI - Determination of reactive oxygen species in nasal polyps. AB - A strong relationship between tissue damage and reactive oxygen species (ROS) has been established by previous studies. The aim of the current study was to evaluate the presence of ROS in nasal polyps (NP) by measuring luminol and lucigenin amplified chemiluminescence (CL). Three groups of specimens were studied. Group 1 composed of NPs of 15 patients, and group 2 constituted of healthy appearing nonpolypoid nasal mucosa of the same patients. Group 3 specimens (control group) were obtained from 15 patients who underwent septoplasty and/or inferior turbinectomy operations, and detected to be free of rhinosinusitis. None of the patients had a history of allergy, asthma, or aspirin sensitivity, and all patients showed negative results to a skin prick test. The ROS levels were measured directly with luminol and lucigenin amplified CL. CL measurements revealed significant differences between ROS constituents of NP (group 1) and control (group 3) tissue samples. CL measurements of healthy appearing nonpolypoid nasal mucosa (group 2) of the NP patients revealed values that were scattered between the values of the other two groups. Although insignificant, ROS levels of the nonpolypoid nasal mucosa (group 2) were found to be higher than normal controls (group 3). In this study, ROS levels of NP tissue samples were directly measured. In our study, by measuring high ROS levels in NP samples, a strong relationship between tissue damage in NPs and ROS has been demonstrated, and the contribution of ROS in the pathophysiology of nasal polyposis has been emphasized. PMID- 16218512 TI - Mucosal immunity of nasopharynx: an experimental study in TCR-transgenic (OVA23 3) mice. AB - The ideal vaccine therapy has been warranted for activation of the mucosal immune response in the upper respiratory tract against various types of microbial infection. However, the precise study in regard to the mucosal route of vaccine administration and its mechanism of action remains to be further investigated. Therefore, to better understand the exact mechanism of nasopharyngeal mucosal immunology, from T-cell aspects, the antigen-specific antibody response was investigated in T cell receptor transgenic (OVA23-3) mice (Tg-mice) and wild type BALB/c mice, in comparison, which were stimulated with repeated nasal antigen challenges of ovalbumin (OVA) together with cholera toxin (CT) or OVA alone. OVA specific IgA and IgG antibodies were not detected in nasal washings of BALB/c mice when these mice were intranasally stimulated with OVA alone. But they were detected in those of BALB/c mice stimulated with OVA and CT, as we have already reported. Interestingly, OVA-specific IgA and IgG antibodies were significantly higher in nasal washings of Tg-mice stimulated with OVA and CT or OVA alone rather than those of BALB/c mice stimulated with OVA and CT. In line with data of the antibody response, OVA-specific IgA and IgG antibody-producing cells significantly increased in number in nasal passage (NP), nasopharyngeal associated lymphoreticular tissue (NALT), cervical lymph node (CLN), and spleen (SP) of these mice. In nasal washings of Tg-mice, interferon (IFN)-gamma and interleukin (IL)-4 was detected even with a small amount of antigen. To see the cytokine profile of NALT, NP, CLN, and SP of these mice, various cytokine concentrations were measured in supernatants of these cells cultured in vitro with OVA. As a result, IFN-gamma was detected at significantly higher levels in culture supernatants of lymphocytes sampled from NP, CLN, SP as well as NALT of mice having increased antibody titers in nasal washings. On the other hand, Th2 type cytokines such as IL-4, IL-6 and IL-13 were efficiently detected in culture supernatants of NP, CLN, and SP cells from Tg-mice mice, but not in those from NALT cells of those mice. All these data taken together indicate that helper T cells recruited into nasal mucosa and locally activated in an antigen-specific fashion, as well as NALT T cells, are essential for mounting local antigen specific antibody responses. PMID- 16218513 TI - Comparison of antigen-induced leukotriene and histamine release from nasal scrapings in allergic rhinitis. AB - BACKGROUND: In the early phase response of allergic rhinitis, the nasal mucosa produces important mediators including histamine and leukotrienes. OBJECTIVE: To investigate the relationship between antigen-induced leukotriene release and histamine secretion in nasal scrapings. METHODS: Using nasal mucosal scrapings from patients sensitized to only house dust mite, we studied the time course of antigen-induced leukotriene release and its relationship to histamine release. RESULTS: Cumulative peptydyl leukotriene (LT) production from nasal scrapings increased from 10 min to approximately 90 min following exposure to mite antigen. The rate of LT release was small (<5 pg/10 min) until 10 min following antigen exposure, increased to approximately 250 to 350 pg LT/10 min from 10 to 45 min post exposure, was reduced to <100 pg/10 min by 60 to 150 min, and by 180 min LT production was negligible. By contrast, histamine secretion began 30 sec after antigen exposure and was complete within approximately 10 min. Net antigen induced LT secretion strongly correlated (R=0.72) with net antigen induced histamine secretion with a ratio of 1:8.7. In addition, net LT/ng histamine and total LT secretion correlated well with antigen-specific IgE in serum, and with the patients' symptoms. CONCLUSION: There is a close relationship between amounts of histamine and LT secretion from antigen challenged nasal mucosa, although the time course of LT release is delayed. In the early phase response, LT are likely to be generated from mucosal mast cells, and thus, mast cell activation will provide an important therapeutic target. PMID- 16218514 TI - Comparison of decongestive capacity of xylometazoline and pseudoephedrine with rhinomanometry and MRI. AB - Topical and systemic sympathicomimetics have been used for many years as a treatment of nasal congestion in diseases such as coryza and sinusitis. The aim of this study was to perform an objective measurement of the decongestive capacity of topical xylometazoline and oral pseudoephedrine in normal subjects and patients with chronic sinusitis. Ten healthy subjects and 10 patients with chronic sinusitis were included in this study. Xylometazoline (0.1%) and pseudoephedrine (120 mg) were each administered to 5 healthy subjects and to 5 patients with chronic sinusitis. Decongestion was measured with active anterior rhinomanometry before, 15 minutes, 30 minutes and 1, 2, 4, 6 and 8 hours after administration. Before and about 90 minutes after drug administration a MRI was performed to visualize the decongestive effect on the turbinates and the mucosa of the sinuses. Xylometazoline reduces the nasal airway resistance for an average 37.3% in all patients and healthy subjects during 8 hours. Pseudoephedrine does not show a clear and long lasting decongestive effect on the turbinates. Important interindividual differences are also noted. MRI clearly shows a clear cut superiority of xylometazoline over pseudoephedrine concerning decongestion of the nasal mucosa. However, there was no decongestive effect whatsoever on the mucosa of the sinuses with either sympathicomimetic. PMID- 16218515 TI - Ethnic variation in sinonasal anatomy on CT-scanning. AB - Computed tomography (CT) scanning of the paranasal sinuses provides valuable information in assessing extent of disease and fine detailed anatomy prior to endoscopic sinus surgery. Awareness of the different anatomic variants of the bony sinonasal anatomy will help the rhinologic surgeon's orientation during the procedure. We conducted a study to look at the incidence of the anatomic variation within the lateral wall of the nose and sinuses and to determine if there is any ethnic difference in these variants between a cohort of 100 Caucasian patients undergoing endoscopic sinus surgery in London and 100 Chinese patients treated in Hong Kong. We compared the two groups with chi-square test and the significant areas are those with p value of less than 0.05. The results show a higher incidence of pneumatisation of the middle turbinate (concha bullosa) and paradoxical bending of the middle turbinate in the Caucasian population. The infraorbital and suprabullar cell development was greater in the Caucasian population though the incidence of sphenoethmoidal cells was much greater in the Chinese population. When asymmetry of the anterior ethmoidal roof was considered, the left was consistently the highest in both groups, though there was no difference in the depth of the cribiform niche between right and left or between Caucasian and Chinese. The incidence of bent uncinate process and of complete absence of a sinus was higher in the Chinese population. There was no difference in the presence of pneumatisation of the agger nasi, of the uncinate process, or of the anterior clinoid process. There was no difference in the presence of septation adjoining the carotid in the lateral wall of sphenoid or in the exposure of the optic nerve within sphenoid or posterior ethmoid sinuses. Although there is no evidence that variants of the sinonasal anatomy seen on CT Scan have a causative effect in the disease process; a knowledge of their presence is paramount in minimising the potential for surgical complications. PMID- 16218516 TI - Nasal cytologies--impact of sampling method, repeated sampling and interobserver variability. AB - The influence of sampling technique, repeated sampling and the interobserver variability on quantitative analysis of nasal cytologies were explored. Nasal cytologies from 12 healthy and 4 allergic volunteers were repeatedly collected with a total of three different sampling techniques. Cells were demonstrated by May-Grunwald-Giemsa staining and by immunostaining for CD45 and eosinophil peroxidase (EPO). To determine the interobserver variability specimens were evaluated by three observers with various levels of nasal cytology experience. When adjusted for the variability due to sampling method and repeated sampling, the staining index for CD45+ cells was 53.4 +/- 11% (interindividual coefficient of variation: 0.21) with an average intraindividual coefficient of variation of 0.14. For CD45, staining indices differed significantly between the washing techniques and the nasal swabs (p<0.01). Repeated sampling did significantly influence the staining indices of specimens collected with nasal swabs (p<0.05). Subjects with nasal allergy had less CD45+ cells in their nasal cytologies than the non-allergic subjects (p=0.02). A high level of compliance for the quantitative analysis of nasal cytologies between three observers was found (kappa(w) range 0.95-0.98). For studies that require repeated sampling, the lavage will be recommended as best suitable to obtain nasal cytologies. PMID- 16218517 TI - Do citation classics in rhinology reflect utility rather than quality? AB - Citation rates have been suggested to be more of an indicator of utility than quality. The aim of this study was to apply measures of utility and quality to articles identified as citation classics in rhinology/anterior skull base surgery. There were 14 articles analysed in the study. The assessment of quality was performed by combining factors from previous publications on quality assessment and the various elements were categorised into four groups; quality of written article and publication, quality of research, quality of evidence-based methodology and quality of outcome. This study revealed that citation classics in rhinology/anterior skull base surgery were well-written and satisfied peer review in reputable journals in the specialty. Quality is satisfied by clarity of exposition and patient numbers. The research was generally asking an important question and the methodology overall was adequate and appropriate for the type of study performed. A good quality of research and outcome was demonstrated with a definite historical importance, and reports that stimulated further research and enquiry. Quality is not satisfied by the lack of randomised controlled trials, appropriate statistical analysis or patient criteria. In conclusion citation rates when considered as an individual measure, reflect utility rather than quality. PMID- 16218518 TI - Isolated maxillary sinus Ewing's sarcoma. AB - Extraskeletal Ewing's sarcoma is rarely found arising in the head and neck region. Only six cases of extraskeletal Ewing's sarcoma invading the nasal cavity or paranasal sinuses have been reported in the literature. The first described case of that Ewing's sarcoma located in isolated maxillary sinus extending into the orbita without involving the nose is presented. A 16-year-old woman presented with swelling in her right cheek. The nasal examination was normal. A computed tomography (CT) and magnetic resonance imaging (MRI) of the sinuses revealed a large mass in the right maxillary sinus with extension to the orbita. Under general anesthesia biopsies were taken. Based on histopathological analysis, a diagnosis of extraskeletal Ewing's sarcoma was made. The patient did not accept the surgical therapy. The patient was treated with combined chemotherapy and radiotherapy. The patient has done without evidence of recurrence or metastasis for one year. PMID- 16218519 TI - Nasal and paranasal sinus manifestation of Rosai-Dorfman disease. AB - Rosai-Dorfman disease is a rare, idiopathic, benign, histiocytic proliferation usually seen in younger patients. Most commonly neck lymph nodes are involved with a predominant infiltration of sinusoidal histiocytes. Nearly half of the patients have extranodal manifestation. Two cases of Rosai-Dorfman disease involving the nose and the paranasal sinuses are presented. The clinical presentation, radiographic findings, treatment and histological characteristics of the disease are discussed. PMID- 16218520 TI - Atrophic rhinitis in a patient with anhidrotic ectodermal dysplasia. AB - We would like to present the rare case of a now 37-year old female patient with autosomal-recessively inherited anhidrotic ectodermal dysplasia being treated in our ENT department for atrophic rhinitis. The clinical appearance very much resembled the picture of an "empty nose" with distinct hypoplasia of the turbinates and extensively wide nasal cavities. We want to point out the possible existence of atrophic rhinitis against the background of an underlying syndromatic disease in adults and also the pediatric patient. PMID- 16218521 TI - European Position Paper on Rhinosinusitis and Nasal Polyps. PMID- 16218522 TI - [Indications for validity of computed tomography and magnetic resonance imaging of the temporomandibular joint]. AB - Recurrent pains of the temporomandibular joint represent a frequent symptom with numerous different causes. CT and MRI can reliably show the cause of these disorders and therefore have substituted conventional X-ray imaging. Modern multi slice-CT (MSCT) allows for examination of the skull base including the mandible in a very short time with thinnest slice collimation (0.75mm). With 2D- and 3D reformations reconstructed out of this volume data set in parasagittal, coronal or any other arbitrary slice orientation excellent imaging of fractures and bony changes of arthrosis as well as benign and malignant tumors of the temporomandibular joint can be performed. MRI offers very good soft tissue contrast in order to visualize the intra-articular disc, the ligaments and muscles, as well the possibility to acquire cross sectional images in any user defined orientation. MRI is the method of choice to diagnose ,,internal derangement", particularly displacement of the intra-articular disc and inflammatory disease of the temporomandibular joint. The present paper will provide diagnostic strategies for the use of MSCT or MRI imaging concerning the different causes of disorders of the temporomandibular joint. PMID- 16218523 TI - [High frequency sonography in the detection of finger injuries in sport climbing]. AB - Sport climbing shows an increasing popularity, including even school sport activity on high climbing levels. Climbing related injuries are predominantly located in fingers/ hands ("climber's finger"), and can present as annular pulley rupture or more common as overuse injury. Beside clinical examination imaging modalities such as MRI and high frequency sonography have shown to allow for detection of climbing related injuries. High frequency sonography enables for exact differential diagnosis of climbing related finger/hand injuries and therefore plays a central role in adequate therapeutic management. PMID- 16218524 TI - [Central pontine myelinolysis with a hyperintense lesion in diffusion weighted MRI: overview by means of a case report]. AB - Central pontine myelinolysis (CPM) is a demyelinating disease of the pons often associated with demyelination of other areas of the central nervous system (CNS). The etiology and pathogenesis of this disorder are still not fully understood. However, almost all cases of CPM are related to severe diseases or chronic alcoholism and occur in the setting of rapidly corrected serum hyponatremia and hypotonicity respectively. Depending on the involvement of other CNS structures, the clinical picture can vary considerably. Magnetic resonance imaging is the most sensitive investigation for the antemortem diagnosis of CPM, although the radiological findings lag behind and do not necessarily correlate with the clinical picture. Quite obviously diffusion-weighted imaging can be useful in the rapid diagnosis of CPM. This short review summarizes the current knowledge on the pathogenesis, clinical presentation, radiological findings, prognosis and therapeutic approaches of CPM. Characteristical clinical features and MR-findings including hyperintensity on diffusion-weighted images are illustrated by a typical case. PMID- 16218525 TI - [Nevoid basal cell carcinoma syndrome]. AB - Nevoid Basal Cell Carcinoma Syndrome (NBCCS) is an autosomal-dominant disorder characterized by multiple basal cell carcinomas, jaw cysts, palmar/plantar pits, calcification of the falx cerebri, and spine and rib anomalies. The combination of clinical, imaging, and histological findings is helpful in identifying NBCCS patients. Imaging plays a crucial role in evaluation of these patients. We present a wide variety of clinical and radiological findings characteristic of this disease. PMID- 16218526 TI - [Long-term follow-up of radical retropubic prostatectomy for prostate cancer in 400 consecutive patients]. AB - PURPOSE: To evaluate the long-term outcomes following radical retropubic prostatectomy for localized prostate cancer. STUDY DESIGN AND METHODS: The charts of 401 patients who underwent radical retropubic prostatectomy between 1992 and 2004 were reviewed. RESULTS: The average age was 63 years with a median PSA of 8.5 and median Gleason score of 5. Ninety one percent of the cases were diagnosed by a single prostate biopsy set (average 7 cores, range 3-16). Pelvic lymph node dissection was performed in 84 patients (21.7%). Nodal involvement was noted in 5 (6%). Capsular invasion was found in 91 patients (24%), positive surgical margin in 78 (20.1%) and seminal vesicle involvement in 37 (9.5%). The average admission stay was 5.2 days (3-22). Fever (11.4%), urine leakage (8%), wound infection (4%) and pulmonary embolism (1.5%) were the most common early complications. One year after the operation 72% of the patients were fully continent. Of the 172 patients who underwent nerve-sparing procedure, 80 patients (46.5%) regained their sexual potency one year after the operation and did not require pharmacological assistance. Median follow-up time was 84 months. The five and eight years overall survival rates were 85% and 73%, respectively. Ten years survival of 136 patients (35%) with clinical stage T1C was 95%. Biochemical failure was observed in 68 patients (18%). The average time to biochemical failure was 12.5 months (range 0 76. median 12). Forty five patients (12%) died, twenty one (5.4%) due to metastatic disease. Mortality correlated to pre-operative PSA, clinical stage and Gleason score. CONCLUSION: At one year follow up, most of the patients are continent and 46.5% have spontaneous erection following nerve-sparing procedure. The overall survival and the long-term outcomes of radical retropubic prostatectomy are equal to those reported in the current literature. PMID- 16218527 TI - [Shock wave lithotripsy for ureteral stones--single institute experience in 661 consecutive cases]. AB - INTRODUCTION: Optimal treatment for ureteral stones is still controversial. The main retreatment options include ureteroscopy and shock wave lithotripsy (ESWL). The study aimed to assess the efficacy of ESWL using the unmodified HM3 lithotripter (Dornier Medical Systems) to achieve stone-free status in patients with ureteral calculi, regardless of stone size and location along the ureter. MATERIAL AND METHODS: We retrospectively reviewed the charts and radiology studies of all patients who had ESWL for ureteral stones. Following ESWL with the HM3 lithotripter, patients had imaging examinations performed after 1 and 3 months, and thereafter, according to the residue status of the stones. RESULTS: During the years 1998-2003, 661 patients underwent ESWL for ureteral stones. The mean age was 51 years (range: 4-85 years) and the average stone size was 8 x 10 mm (range: 3-30 mm). Forty percent of the patients had upper ureteral calculi, 17% mid and 43% lower ureteral stone. Overall stone-free status was 93.6%. According to stone location, the success rate for upper ureteral calculi was 93.6%, and 93.6% and 95.5% for mid and lower ureteral stones. Complications were recorded in 28 (4.2%) patients, 20 had acute urinary tract infection treated with i.v. antibiotics, and 8 had renal colic treated conservatively. CONCLUSIONS: Our data shows that ESWL using HM3 lithotripter for ureteral stones at any level and size has a high success rate with minimal morbidity. ESWL using the HM3 lithotripter is our preferred treatment of choice for ureteral calculi. PMID- 16218528 TI - [The laparoscopic approach to renal tumors outcome of 121 laparoscopic radical and partial nephrectomy procedures]. AB - BACKGROUND AND PURPOSE: We present and review a single center experience with laparoscopic renal surgery for renal cancer including laparoscopic radical and partial nephrectomy. PATIENTS AND METHODS: During the period September 2002 - January 2005, 180 patients underwent laparoscopic renal surgery at our center for several indications. A total of 121 patients underwent radical nephrectomy (76) or partial nephrectomy (45) for solid renal tumors. Their data was recorded and analyzed including age, indication for surgery, operative time, blood loss, intra and postoperative complications, conversion rates, histological results and outpatient follow-up. RESULTS: The mean age at surgery was 65 years (range: 21-89 years). The indications for surgery were solid, enhancing renal masses. For tumors larger than 4 cm, a radical nephrectomy was performed. Tumors that were smaller than 4 cm were removed by partial nephrectomy. The mean operative time was minutes (range: 70-280 minutes) and the mean blood loss was 220 cc (range: 30 700 cc). The mean blood loss was significantly lower in the radical nephrectomy group (120 cc) than in the partial nephrectomy group (320 cc). In eight patients major intraoperative complications were noted including injury of the hilar vessels (5 cases), inferior vena cava (1 case), large bowel and spleen (one patient) and duodenum (1 case). Conversion to open surgery was needed in nine patients (7.4%), four in the radical nephrectomy and five in the partial nephrectomy group. Major postoperative complications were recorded in two patients who underwent radical nephrectomy (pneumothorax and incarcerated inguinal hernia) and in two patients who underwent partial nephrectomy (urine leakage). The pathological examination demonstrated renal cell carcinoma in 109 cases, oncocytoma in 6 cases, angiomyolipoma in 2 cases, sarcoma of the kidney and metastasis from lung cancer in one case each and a hemorrhagic cyst in one case. In all patients who underwent radical nephrectomy negative surgical margins were obtained, in three patients after partial nephrectomy the surgical margins were focally involved by tumor. The mean tumor size was 5.1 cm and 3.1 cm after radical or partial nephrectomy respectively. During follow-up, none of the patients developed local or distant recurrence. CONCLUSIONS: The laparoscopic approach to kidney cancer seems to be safe and oncologically sound. The low morbidity rate together with the inherent advantages of laparoscopic surgery make this approach attractive and we believe it should be considered the new standard of care for renal cancer. PMID- 16218529 TI - [Endoscopic injection of dextranomer/hyaluronic acid for vesico-ureteral reflux- preliminary results]. AB - BACKGROUND: Endoscopic injection is a minimal invasive treatment for vesico ureteral reflux, an alternative for prophylactic antibiotics or open surgery. Concerns of safety were raised regarding traditional injectable materials used in the past. The recently introduced dextranomer/hyaluronic acid is a new biocompatable substance. Its safety and efficacy has been investigated and demonstrated. OBJECTIVES: To report our preliminary results with the endoscopic injection of dextranomer/hyaluronic acid. METHODS: Twenty nine pediatric patients of mean age 58 months (range 6-144) underwent endoscopic treatment. A total of 42 refluxing ureteral units were treated. Reflux grade was 2, 3, and 4 in 10, 24, and 8, respectively. The procedure was performed under general anesthesia, using a 9.5FR pediatric cystoscope and a semi-rigid needle. The substance was injected submucosally at the 6-o'clock position of the ureteral orifice. A mean of 0.87 ml of the substance was injected (range: 0.6-1.2). In the latter part of the series we injected a minimum of 1 ml for each ureter. During follow-up, ultrasonography and a voiding cystography were performed after 1 and 3 months respectively. RESULTS: Vesico-ureteral reflux resolved in 32/42 (76%) of the ureters. The rate of success was 10/10 (100%), 19/24 (79%) and 4/8 (50%) in reflux grades 2, 3, and 4, respectively. De-novo contralateral reflux was noted in 5 patients. A single complication occurred: acute venous bleeding originating in the injection site, which was managed endoscopically. CONCLUSIONS: Endoscopic injection of dextranomer/ hyaluronic acid is a safe and effective treatment. We recommend the injection of 1 ml, the entire volume available for each ureter. PMID- 16218530 TI - [Endourological treatment of ureteropelvic obstruction using holmium YAG laser]. AB - BACKGROUND: Ureteropelvic junction obstruction (UPJO) is defined as an impairment of urine outflow from the pelvis to the ureter. Several treatment approaches are in use. OBJECTIVES: To present the different endo-urological approaches- retrograde and antegrade techniques --for UPJO using the holmium: YAG laser. METHODS: We followed 67 patients who underwent endopyelotomy between 1994 - 1997 and compared the different approaches for the treatment of UPJO. RESULTS: A total of 67 patients were operated, 52 patients using retrograde approach and 15 using antegrade approach. In the retrograde approach success was achieved in 48 patients (93%) and in 14 patients (94%) in the antegrade approach. Complications rate was 10% and 40%, respectively. Hospital stay was 2.2 days in the retrograde approach and 4.6 in the antegrade approach. CONCLUSIONS: The success rate in the different endourologic approaches was high and no difference between the two approaches was identified. From our experience, lower complication rate and shorter hospital stay make the retrograde approach more advantageous. PMID- 16218531 TI - [Nephron sparing surgery as a treatment modality for renal angiomyolipoma]. AB - OBJECTIVE: There is no consensus concerning the indications for active treatment (selective angioembolization (SAE) or surgery) versus observation alone, and the treatment modality for patients with renal angiomyolipoma (AML). Our study aimed to analyze the results of long-term follow-up of patients with renal AML treated by nephron-sparing surgery (NSS). MATERIALS AND METHODS: The medical records of thirteen patients with renal AML were retrospectively reviewed for clinical and histopathologic features. There were twelve women and one man (mean age 55 years) who were treated by NSS in our medical center between the years 1993-2001. RESULTS: One patient with tuberous sclerosis had bilateral multifocal tumors, another patient that presented with shock and retroperitoneal bleeding underwent an urgent NSS. Overall, eight patients were symptomatic at diagnosis. The mean tumor diameter was 5.5 cm (range 2.5-13 cm). There was no need to perform nephrectomy in any of the patients. Two patients had enucleation of more than one tumor in the same session. In one case the adrenal gland was surrounded by the AML and was removed with the specimen. No other serious complications were recorded. Four patients required blood transfusions. The mean follow-up period was 26 months (range: 6-62 months) and during that time there was no evidence of recurrence or decreased renal function. Only one patient developed AML in the contralateral kidney, there was one case of incisional hernia and another patient continued to suffer from flank pain. CONCLUSIONS: The results of the present study suggest that NSS is a safe and effective treatment modality for renal AML. PMID- 16218532 TI - [Tissue microarray--an infrastructure for prostate cancer outcome research]. AB - In tissue microarray (TMA) approximately 500 clinical formalin fixed and paraffin embedded tissue cores are reorganized into a new recipient block. Each recipient block may be sectioned up to 200 times. MATERIAL AND METHODS: Radical prostatectomy specimens of 405 patients operated for prostate cancer (CaP) between 1992 and 2004 were considered. A large database was created incorporating clinical and pathological data. Median follow-up time was 84 months. A full pathological revision was performed. We allocated and marked sites of interest on patients' slides: CaP, normal prostate, capsular or seminal vesicles invasion and the appropriate controls. The corresponding paraffin blocks served as donor blocks for harvesting tissue cores sequentially incorporated into the TMA recipient block. RESULTS: The specimens of 264 patients out of 405 qualified for inclusion into the TMA (yield 65%). Two TMA copies were prepared containing 4 blocks each. Each copy includes approximately 2000 tissue cores. CONCLUSIONS: We have constructed a large scale TMA associated with a detailed clinical database. This TMA will serve for prostate cancer outcome studies by using immunohistochemical as well as other molecular staining. Our TMA is unique because it contains sites of interest for CaP local invasiveness and metastasis. PMID- 16218533 TI - [Does percutaneous nephrolithotripsy (PCNL) affect renal function: assessment with quantitative spect of Tc 99M-DMSA (QDMSA) renal scintigraphy]. AB - PURPOSE: The study aimed to quantitatively investigate the effect of PCNL on global and regional function using quantitative single photon emission computerized tomography (SPECT) measurement of Tc-dimercaptosuccinic acid (DMSA) uptake by the kidneys (QDMSA). METHODS: Seventy nine patients with nephrolithiasis undergoing PCNL were studied by sequential QDMSA examination. There were 42 (53%) males and 37 (47%) females with mean age of 47 +/- 16 years. The initial study was conducted before PCNL procedure and the follow-up studies were performed 1.5-24 months after PCNL. Among 60 of the 79 (76%) patients, in whom PCNL was performed using upper or lower pole access, regional renal function of affected and non-affected pole of operated kidney was calculated separately. RESULTS: There was no statistically significant difference between the uptake of treated kidney before and after PCNL procedure (11.9% +/- 5% vs 11.6% +/- 5%; t = 0.9, p = 0.368). The total renal functional volume of treated kidney was to be slightly decreased from 235cc +/- 62cc to 224cc +/- 59cc (t = 2.7; p = 0.011). The percent of injected dose per ml.. of renal tissue of treated kidney was not affected statistically (0.051 +/- 0.02 vs 0.053 +/- 0.02; t = 0.86, p = 0.296). In the assessment of the regional renal function of treated kidney, a statistically significant decrease in the functional renal volume was revealed at the part which underwent PCNL procedure (91cc +/- 30cc vs 82cc +/- 27cc; t = 2.64, p = 0.013). Regarding percent of injected dose per ml. of renal tissue, no statistically significant difference was found between the part of treated kidney, which underwent PCNL and non-affected area of the same kidney (0.049 +/- 0.02 vs 0.05 +/- 0.02; t = 0.693, p = 0.494). The function of contralateral kidney remained unchanged (13.4% +/- 5.2% vs 13.6% +/- 4.8%; t = 0.68, p = 0.5). Function volume, neither total percent uptake, nor percent of injected dose per ml. of renal tissue were reduced significantly. Further studies with long term follow-up of treated kidney are required. PMID- 16218534 TI - [Detection of sexually transmitted pathogens in patients with hematospermia]. AB - BACKGROUND: Although the current literature attributes most cases of hematospermia to an infectious agent, identification of the specific pathogens involved has been limited. OBJECTIVE: To determine the prevalence of different pathogens, in patients presenting with hematospermia to our sexually transmitted disease clinic. PATIENTS AND METHODS: We retrospectively examined the laboratory results of 30 patients who presented to our clinic with hematospermia after other non-infectious pathologies had been excluded by a referring physician. Firstly, void urine samples as well as genitourinary and serum specimens were tested for Herpes simplex, Chlamydia trachomatis and Ureaplasma urealyticum. Standard bacterial cultures were also performed. RESULTS: In 12 of the 18 patients a possible pathogenic cause for the hematospermia was found. Evidence for Herpes simplex virus (Type 2) was found in 5 of the patients (16.6%), for Chlamydia trachomatis in 3 patients (10%) and for Ureaplasma urealyticum in 2 of the patients. Enterococcus fecalis was detected in 2 (6.6%) of the patients and in 1 patient Herpes simplex virus (Type 1) antigen and antibodies were detected. In all cases in which evidence of a pathogen was identified, the appropriate antimicrobial agent was administered, with recurrence in two patients only. CONCLUSIONS: Recent advances in microbiologic diagnostic techniques have facilitated the detection of pathogens in patients with hematospermia, thereby enhancing the efficacy of treatment. PMID- 16218535 TI - [Trends of electronic publishing in medicine and life sciences]. AB - Scientific publication in the electronic media is gaining popularity in academic libraries, research institutions and commercial organizations. The electronic journal may shorten the processes of writing and publication, decrease publication and distribution costs, and enable access from any location in the world. Electronic publications have unique advantages: it is possible to search them, to create hyperlinks to references and footnotes, as well as to information on the web and to include graphics and photographs at a very low cost. Audio, video and tri-dimensional images may also be included. Electronic publishing may also speed up review and publication processes and enable the writer to receive immediate feedback through the web. However, in spite of the advantages, there are certain points that must be considered: accessibility to previously published material is not guaranteed as databases are not always stable and coverage may change without notice. In addition, the price that commercial publishers charge for their services may be very high or be subject to the purchase of a packaged deal that may include unwanted databases. Many issues of copyright and the use of published material are not yet finalized. In this review we discuss the advantages and disadvantages of the electronic scientific publication, the feasibility of keeping appropriate quality and peer-review process, the stability and accessibility of databases managed by the publishers and the acceptance of the electronic format by scientists and clinicians. PMID- 16218536 TI - [Dermatological manifestations in the antiphospholipid syndrome]. AB - Antiphospholipid syndrome is regarded as a systemic disease. A total of 41% of the patients are presented with one form of the diverse skin findings at diagnosis. Due to the high prevalence of skin affliction it was recently suggested to include this syndrome in the diagnostic criteria of APS. The importance of these findings also entails the fact that in 25% of the patients the skin manifestations precede thromboembolic phenomena. The findings entail livedo reticularis, anetoderma, skin necrosis and others. This article discusses their prevalence and their relationships to other clinical manifestations in APS and how to approach them therapeutically. PMID- 16218537 TI - [New usage for an old drug: acetylcysteine for contrast-induced nephropathy]. AB - Contrast induced nephropathy is the third common cause of hospital-acquired renal failure. The pathophysiology and the mechanism involved in the contrast media induced renal failure are not well understood but it is thought to be the result of direct renal toxicity and medullary ischemia. The only preventive measure proven to reduce such nephropathy is hydration (pre- and post-procedure). However, the above treatment is partially effective. The antioxidant N acetylcysteine has recently been studied for the prevention of contrast induced nephropathy in several prospective, randomized, placebo-controlled studies among patients with moderate renal insufficiency. Although some discrepancies exist, N acetylcysteine appears to reduce the risk of contrast-induced nephropathy. N acetylcysteine has a safety profile and low cost. Taken together, N acetylcysteine is recommended for the prevention of contrast-induced nephropathy in high-risk patients. PMID- 16218538 TI - [Guidelines for the prevention and treatment of atherosclerosis and cardiovasculer diseases: treatment of diabetes mellitus, dyslipidemia and the prevention of stroke]. AB - Atherosclerosis is one of the principal causes of morbidity and mortality worldwide. The recent decades have witnessed great advances both in the identification of risk factors for the development of atherosclerosis and the treatment of its complications. This effort was rewarded with the reduction of mortality rates from cardiovascular diseases. The need for an update of the recommendations for the prevention of atherosclerosis and cardiovascular diseases stems from a large body of recently published trials, leading to fundamental changes in the way we treat patients with various levels of risk. The second part of the guidelines deals with the treatment of diabetes mellitus, dyslipidemia and the prevention of stroke. PMID- 16218539 TI - [ADMA (asymmetric dimethylarginine)--the inhibitor of nitric oxide (NO) synthesis: a new marker for vascular pathology]. AB - Ample evidence is accumulating to suggest that asymmetric dimethylarginine (ADMA), an endogenous competitive inhibitor of nitric oxide synthase, is significantly elevated during phases of endothelial dysfunction. ADMA inhibits NO synthesis, hence its arterial infusion induces local arterial constriction. ADMA is generated ubiquitously in numerous tissues, by proteolysis of methylated proteins, while its degeneration is carried out mainly by the enzyme dimethylarginine dimethylaminohydrolase (DDAH). Administration of L-arginine can override partially NO synthesis by ADMA, yet it cannot eliminate the primary factors involved in the endothelial dysfunction. ADMA measurements might add valuable information about this new risk factor or at least a marker for adverse endothelial events. PMID- 16218540 TI - [Prophecy and mental illness]. AB - It is a well known platitude that a mentally ill person may "think that he is God" or "believes that he is the Messiah". Despite the generalization and shallowness of this attitude, sometimes psychotic patients indeed have delusions with contents of divine revelation, messianic assignments or prophetic power. In this current article we examine the different connections between prophecy and mental condition, especially psychotic. We present sources that combine prophecy and insanity, and also possible psychiatric interpretation of these situations. Finally, we present the attitude of the Rambam to prophecy and the personality characteristics of the prophet, limiting the possibility of the mentally ill patient who pretends to be a prophet. PMID- 16218541 TI - [In Hebrew, please say sensitive bowel syndrome instead of irritable bowel syndrome]. PMID- 16218542 TI - CMS' changes to IPPS will affect your hospital's reimbursements. PMID- 16218543 TI - Ensure quality indicators are documented. PMID- 16218544 TI - Clinical pathways can help you track data. PMID- 16218545 TI - Pathway keeps LOS low for young diabetes patients. PMID- 16218546 TI - Patient safety alert. HCA nears completion of systemwide eMAR. PMID- 16218547 TI - Hospitals collaborate to reduce surgical infections. PMID- 16218548 TI - Early '100,000 Lives' participant sees benefits. PMID- 16218549 TI - Discharge planning advisor. Innovative program helps homeless, frees up beds. PMID- 16218550 TI - Sutter Health seeks dialysis solutions. PMID- 16218552 TI - The knee: breaking the MR reflex. PMID- 16218553 TI - Foot and ankle disorders: radiographic signs. PMID- 16218554 TI - Variants of exostosis of the bone in children. PMID- 16218557 TI - Crocker, not Armit and Robinson, begat the six aromatic electrons. PMID- 16218555 TI - A compartmental approach to the radiographic evaluation of soft-tissue calcifications. AB - Musculoskeletal radiology, like most areas of diagnostic radiology, has seen a dramatic increase in the use of cross-sectional imaging techniques over the last decade. However, conventional radiography remains a vital role in the evaluation of disease involving the spine and extremities. Given its widespread availability and low cost, radiography should be the initial examination obtained when imaging is clinically indicated. Therefore it is essential that the practicing radiologists be able to maximize the diagnostic value of this common study to aid the clinicians with a focused differential diagnosis, if not a definitive diagnosis. While much emphasis has been placed on radiographic interpretation of osseous lesions, similar diagnostic yield can be often obtained about calcified or ossified soft-tissue lesions. Detailed evaluation of the distribution and morphology of the soft-tissue calcifications, combined with a thorough knowledge of the entities that may occur at the site of the noted abnormality, provides significant interpretive value to provide a definitive diagnosis or accurately recommend the next most effective management step. We provide a compartmental based approach to the interpretation of soft-tissue calcifications. PMID- 16218558 TI - Conjugated polymers and aromaticity. PMID- 16218559 TI - Sigma- and pi-electron delocalization: focus on substituent effects. PMID- 16218560 TI - Interrelation between H-bond and Pi-electron delocalization. PMID- 16218561 TI - Tautomeric equilibria in relation to pi-electron delocalization. PMID- 16218562 TI - Spherical aromaticity: recent work on fullerenes, polyhedral boranes, and related structures. PMID- 16218563 TI - Curved pi-conjugation, aromaticity, and the related chemistry of small fullerenes (< C60) and single-walled carbon nanotubes. PMID- 16218564 TI - Mobius aromaticity and delocalization. PMID- 16218565 TI - All-metal aromaticity and antiaromaticity. PMID- 16218566 TI - Anisotropy of the induced current density (ACID), a general method to quantify and visualize electronic delocalization. PMID- 16218567 TI - Energetic aspects of cyclic pi-electron delocalization: evaluation of the methods of estimating aromatic stabilization energies. PMID- 16218568 TI - Description of electron delocalization via the analysis of molecular fields. PMID- 16218570 TI - The magnetic shielding function of molecules and pi-electron delocalization. PMID- 16218569 TI - Nucleus-independent chemical shifts (NICS) as an aromaticity criterion. PMID- 16218571 TI - Theoretical evaluation of electron delocalization in aromatic molecules by means of atoms in molecules (AIM) and electron localization function (ELF) topological approaches. PMID- 16218572 TI - Enzymatic fabrication of DNA nanostructures: extension of a self-assembled oligonucleotide monolayer on gold arrays. AB - Nucleic acid nanostructures are useful as templates for bionanofabrication of composite molecular nanostructures in materials science, molecular electronics, and biosensing. Here, we demonstrate that terminal deoxynucleotidyl transferase, which repetitively adds mononucleotides to the 3' end of a short DNA initiator, can be used to rapidly fabricate DNA nanostructures up to 121 nm high with lateral dimensions from 0.1 to 4 mum in 2 h. These programmable scaffolds can potentially be employed to build more complex nanostructures consisting of natural or unnatural nucleotides with selective docking sites along the single stranded DNA. PMID- 16218573 TI - Dispersing nanotubes with surfactants: a microscopic statistical mechanical analysis. AB - A first theoretical study of surfactant-stabilized carbon nanotube dispersions is presented. Density functional theory is used to compute potential of mean force between nanotubes in an aqueous solution of cationic surfactant n decyltrimethylammonium chloride. In agreement with experimental results, it is found that stable dispersions can be prepared for surfactant bulk concentrations below the critical micelle concentration. Computed density profiles of head and tail segments indicate that surfactants adsorb on nanotube surfaces in a random fashion rather than form cylindrical micelles, which is also in agreement with recent small-angle neutron scattering measurements. PMID- 16218574 TI - Revisiting noncovalent So2- amine chemistry: an indicator-displacement assay for colorimetric detection of So2. AB - A supramolecular approach for potential detection of SO2 is presented, which is based on the "old" donor-acceptor chemistry between SO2 and amines and includes an indicator-displacement assay. When amines were added to Zn tetraphenylporphyrin 1 in CHCl3, the solution changed from red to dark green. A bathochromic shift of Deltalambda approximately 10 nm was observed for the Soret band, indicating the formation of 1*amine complexes. After this, SO2 gas was introduced, and the original red color of the solution was restored. The Soret band returned to its position for free porphyrin 1. The 1*amine complexes dissociated, and new SO2*amine adducts formed. Porphyrin 1 thus served as an indirect colorimetric indicator for SO2. The system discriminates between SO2 and such typical exhaust gases as COX, NOX, and H2O. From the indicator-displacement assay, the Kassoc values between 1000 and 30 000 M-1 for SO2*amine complexes were determined, which are comparable to those obtained by direct titration experiments between SO2 and the amines. Spectroscopic features of SO2*amine complexes are also presented. PMID- 16218575 TI - In vitro reconstitution of EryCIII activity for the preparation of unnatural macrolides. AB - EryCIII is a desosaminyltransferase that converts an inactive macrolide precursor to a biologically active antibiotic. It may have potential for the synthesis of unnatural macrolides with useful biological activities. However, it has been difficult to reconstitute the activity of EryCIII in vitro. We report here that purified, inactive EryCIII can be converted to an active catalyst by the addition of another protein encoded in the same gene cluster, EryCII. The EryCII-treated protein retains activity even when EryCII is removed. We also show that AknT, an activator protein from an unrelated gene cluster, is capable of activating EryCIII. Although the mechanism of activation is not yet understood, we have concluded from these experiments that these antibiotic Gtf activator proteins do not function to deliver substrates to EryCIII and do not exert their effects by forming stable complexes with the Gtf during the glycosyltransfer reaction. We report that activated EryCIII is capable of utilizing an alternative sugar donor, so these results lay the groundwork for the production of novel macrolides. PMID- 16218576 TI - Completely geometrically optimized DFT/ONIOM triple-helical collagen-like structures containing the ProProGly, ProProAla, ProProDAla, and ProProDSer triads. AB - We report completely optimized ONIOM DFT/AM1 molecular orbital calculations on several collagen-like triple helices based upon the repeating triad, ProProGly. The requirement of Gly as every third amino acid in collagen can be attributed to its enantiomorphic nature, as it behaves as a Damino acid in collagen. We, therefore, explored related collagen-like triple helices with one of the central Gly's mutated to either L or DAla; l-Ala appreciably destabilizes, while d-Ala slightly stabilizes the triple helical structure. Mutation of the same Gly to DSer, which is simply DAla with an OH in place of one of the methyl H's, induces a much greater stabilization due to an additional H-bond formed between this OH and a C=O on an adjacent peptide strand. Energies are presented for the triple helices and their component strands (both optimized and distorted to their triple helical geometries) relative to the component amino acids. The variation of relative energies with the chosen reference is delineated. PMID- 16218577 TI - Enzyme redesign: two mutations cooperate to convert cycloartenol synthase into an accurate lanosterol synthase. AB - Efforts to modify the catalytic specificity of enzymes consistently show that it is easier to broaden the substrate or product specificity of an accurate enzyme than to restrict the selectivity of one that is promiscuous. Described herein are experiments in which cycloartenol synthase was redesigned to become a highly accurate lanosterol synthase. Several single mutants have been described that modify the catalytic specificity of cycloartenol to form some lanosterol. Modeling studies were undertaken to identify combinations of mutations that cooperate to decrease the formation of products other than lanosterol. A double mutant was constructed and characterized and was shown to cyclize oxidosqualene accurately to lanosterol (99%). This catalytic change entailed both relocating polarity with a His477Asn mutation and modifying steric constraints with an Ile481Val mutation. PMID- 16218578 TI - Separate metal requirements for loop interactions and catalysis in the extended hammerhead ribozyme. AB - An extended hammerhead ribozyme derived from Schistosoma mansoni, including conserved loops in stems I and II, has been examined to directly monitor the relationship between docking of loops and its activity using site-directed spin labeling (SDSL) and EPR spectroscopy. Dynamics with EPR spectroscopy and fast quench kinetics measurements have shown that the docking of stems I and II occurs at low Mg2+ concentrations ([Mg2+]1/2,dock = 0.7 mM, 0.1 M NaCl), but a much weaker Mg2+ interaction ([Mg2+]1/2,cat approximately 90 mM) increases activity to very high maximum rates of approximately 1 s-1 at 0.1 M Na+ and pH 7.0. PMID- 16218579 TI - Methyltransferase-directed DNA strand scission. AB - We demonstrate here that MTase-modified DNA can undergo the Staudinger ligation with triarylphosphines derivatized with phenanthroline. Presentation of these duplexes with Cu(II) and 3-mercaptopropionic acid leads to strand scission proximal to the MTase recognition site. By virtue of their ability to use a synthetic azide-bearing cofactor, M.TaqI and M.HhaI produce a DNA lesion that induces scission 5' to the base modified by the enzyme. This chemistry represents a new approach by which regions of DNA methylation can be rapidly identified on the basis of DNA damage. PMID- 16218580 TI - A supramolecular fluorescence sensor for pyrovanadate as a functional model of vanadium haloperoxidase. AB - A novel basket-shaped tris(pyrene guanidinium) receptor was synthesized which binds pyrovanadate and pyrophosphate with Ka > 107 M-1. The binding of both anions is associated with quenching of the excimer fluorescence of the pyrenes. The supramolecular vanadate complex catalyzes the bromination of activated C-H bonds and hence is an enzyme mimic of vanadium haloperoxidases. PMID- 16218581 TI - Morphogenesis of highly ordered mixed-valent mesoporous molybdenum oxides. AB - Control of the size, shape, and structure of mesoporous transition metal oxide materials is important in their correlations with corresponding optoelectronic and photocatalytic properties. Highly ordered cubic phases of mixed-valent mesoporous molybdenum oxides have been prepared by the reduction and decomposition of aqueous molybdenum precursor solution in the presence of poly(ethylene oxide) under ultrasonic irradiation. Large-scale uniform molybdenum oxide particles with well-defined crystal-like morphologies (ball-like, rhombic dodecahedral, and cubic shapes) were synthesized and found to be controllable by modifying the molecular chain length of the polymeric additive. Molybdenum oxides with an average oxidation state of 4.8 form a cubic lattice of open mesoporous structures. PMID- 16218582 TI - A general route for post-translational cyclization of mRNA display libraries. AB - Cyclic peptides are attractive scaffolds for the design of conformationally constrained molecular therapeutics. Previously, biological display libraries could only be cyclized via disulfide bonds, which are labile and can be reduced in an intracellular environment. In this paper, we construct high diversity, covalently cyclized mRNA display libraries (>1013 sequences) and analyze the cyclization reaction using MALDI-TOF MS and unnatural amino acid incorporation. Our route allows the extent of cyclization to be evaluated quantitatively and is broadly applicable to a variety of cyclization chemistries. PMID- 16218583 TI - Photophysical properties of [Ru(phen)2(dppz)]2+ intercalated into DNA: an integrated Car-Parrinello and TDDFT study. AB - Combined Car-Parrinello and time-dependent DFT calculations performed on [Ru(phen)2dppz]2+ intercalated into an adenine-thymine tetramer reveal a remarkable influence of the base pairs in determining the electronic structure and the character of the excited states involved in the absorption and emission processes. PMID- 16218584 TI - Enhancing 2-iodoxybenzoic acid reactivity by exploiting a hypervalent twist. AB - A rearrangement of hypervalent bonds, or twisting, proves to be the rate determining step in the 2-iodoxybenzoic acid (IBX) oxidation of alcohols. From this insight, derived from density functional theory calculations, we explain why IBX oxidizes large alcohols faster than small ones and propose a modification to the reagent predicted to make it more active. PMID- 16218585 TI - Potent, selective pyrone-based inhibitors of stromelysin-1. AB - In an effort to develop alternatives to hydroxamate-based matrix metalloproteinase inhibitors (MPIs), we have utilized the drug discovery program LUDI enhanced with the structural coordinates of a bioinorganic model complex. This method has yielded the first pyrone-based MPIs. The inhibitors demonstrate nanomolar potency against MMP-3 and are selective for MMP-3 over MMP-2 and MMP-1. We postulate that the potency and unusual selectivity profile of these MPI is attributable to the pyrone chelating group. PMID- 16218586 TI - Selective dye-labeling of newly synthesized proteins in bacterial cells. AB - We describe fluorescence labeling of newly synthesized proteins in Escherichia coli cells by means of Cu(I)-catalyzed cycloaddition between alkynyl amino acid side chains and the fluorogenic dye 3-azido-7-hydroxycoumarin. The method involves co-translational labeling of proteins by the non-natural amino acids homopropargylglycine (Hpg) or ethynylphenylalanine (Eth) followed by treatment with the dye. As a demonstration, the model protein barstar was expressed and treated overnight with Cu(I) and 3-azido-7-hydroxycoumarin. Examination of treated cells by confocal microscopy revealed that strong fluorescence enhancement was observed only for alkynyl-barstar treated with Cu(I) and the reactive dye. The cellular fluorescence was punctate, and gel electrophoresis confirmed that labeled barstar was localized in inclusion bodies. Other proteins showed little fluorescence. Examination of treated cells by fluorimetry demonstrated that cultures supplemented with Eth or Hpg showed an 8- to 14-fold enhancement in fluorescence intensity after labeling. Addition of a protein synthesis inhibitor reduced the emission intensity to levels slightly above background, confirming selective labeling of newly synthesized proteins in the bacterial cell. PMID- 16218587 TI - Cross dipole stacking in the crystal of distyrylbenzene derivative: the approach toward high solid-state luminescence efficiency. AB - Through adding two substituent phenyl groups on distyrylbenzene, we have obtained the cross stacking of 2,5-diphenyl-1,4-distyrylbenzene with two trans double bonds (trans-DPDSB) in crystalline state. In such a cross-stacking mode, the solid-state emission exhibits high-intensity, having characteristics similar to its single molecule. The organic light-emiiting diodes (OLEDs) with attractive performance have been achieved using trans-DPDSB as a light-emitting layer, and the amplified spontaneous emission of the needlelike crystals has been observed. PMID- 16218588 TI - Preparation of fullerene-polyyne nanospheres via thermally induced solid-state polymerization. AB - A 1,3,5,7-octatetrayne-linked bis(fullerene) compound has been synthesized. Through a thermally induced solid-state polymerization reaction on a surface, the solid thin film of this compound was transformed into a highly uniform and well organized polymer nanosphere array. This finding suggests a simple and useful method for the preparation of carbon-rich, fullerene-containing nanoparticles. PMID- 16218589 TI - Narrow pore-diameter polypyrrole nanotubes. AB - Bulk quantities of electrically conducting nanotubes of polypyrrole having narrow pore diameter (6 nm) can be synthesized rapidly by chemical oxidative polymerization of pyrrole in the presence of stoichiometric amounts of V2O5 nanofibers. The V2O5 nanofibers act as templates for polymerization and yield, as the initial product, polypyrrole nanotubes with pores filled with V2O5. The V2O5 dissolves readily in aq. 1.0 M HCl, yielding hollow polypyrrole nanotubes having conductivity of approximately 2 S/cm. As-synthesized polypyrrole nanotubes spontaneously reduce noble metal ions to the corresponding metal nanoparticles at room temperature without any capping or dispersing agents. For example, 3-5 nm size nanoparticles of Ag, Au, and Pd, etc., deposit readily on the surface of the tubes which then migrate spontaneously to the pore, and, in the case of Ag, coalesce in the core, yielding 4-8 nm diameter coaxial cables of Ag surrounded by a 20-30 nm thick polypyrrole fiber sheath. PMID- 16218590 TI - Synthesis of functionalized asymmetric star polymers containing conductive polyacetylene segments by living anionic polymerization. AB - Novel 3-arm ABC, 4-arm ABCD, and 5-arm ABCDE asymmetric star polymers comprising the conductive polyacetylene precursor, poly(4-methylphenyl vinyl sulfoxide) (PMePVSO), and other segments, such as polystyrene, poly(alpha-methylstyrene), poly(4-methoxystyrene), poly(4-trimethylsilylstyrene), and poly(4-methylstyrene), were synthesized by the methodology based on living anionic polymerization using DPE-functionalized polymers. This methodology involves the addition reaction of a DPE-functionalized polymer to a living anionic polymer followed by the living anionic polymerization of MePVSO initiated from the in situ formed polymer anion with two, three, or four polymer segments. The resultant asymmetric star polymers possessed predetermined molecular weights, narrow molecular weight distributions (Mw/Mn < 1.03), and desired compositions as confirmed by SEC, 1H NMR, SLS, and elemental analysis. After thermal treatment, the PMePVSO segment in the star polymer could be completely converted into a conductive polyacetylene segment, evident from TGA and elemental analysis. These asymmetric star polymers are expected to exhibit interesting solution properties and unique microphase separated morphological suprastructures with potential applications in nanoscopic conductive materials. Moreover, this methodology can afford the target asymmetric star polymers with arm segments varying in a wide range and enables the synthesis of more complex macromolecular architectures. PMID- 16218591 TI - Design and construction of a 2D metal organic framework with multiple cavities: a nonregular net with a paracyclophane that codes for multiply fused nodes. AB - A metal organic framework with two different nodes (circle and square) and a structure related to one of the 20 known 2-uniform nets has been constructed using an organic building unit that codes for multiply fused nodes. PMID- 16218592 TI - Temperature- or guest-induced drastic single-crystal-to-single-crystal transformations of a nanoporous coordination polymer. AB - Aside from a temperature-induced structural transformation, a new nanoporous coordination polymer [Ag6Cl(atz)4].OH.6H2O (Hatz = 3-amino-1,2,4,-triazole) comprised of interpenetrated Ag3(atz)2 coordination nets exhibits a single crystal-to-single-crystal transformation of a 5-fold to 6-fold net interpenetration change triggered by guest desorption/absorption. PMID- 16218593 TI - An NMR determination of CO diffusion on platinum electrocatalysts. AB - We report the first direct measurement of CO diffusion on nanoparticle Pt electrocatalysts at the solid/liquid interface, carried out using 13C nuclear magnetic resonance (NMR) with a spin-labeling pulse sequence. Diffusion parameters were measured in the temperature range of 253-293 K for CO adsorbed on commercial Pt-black under saturation coverage. 2H NMR of the same system indicates that the electrolyte remains in the liquid state at temperatures where the CO diffusion experiments were performed. The CO diffusion parameters follow typical Arrhenius behavior with an activation energy of 6.0 +/- 0.4 kcal/mol and a pre-exponential factor of (1.1 +/- 0.6) x 10-8 cm2/s. Exchange between different CO populations, driven by a chemical potential gradient, is suggested to be the main mechanism for CO diffusion. The presence of the electrolyte medium considerably slows down the diffusion of CO as compared to that seen on surfaces of bulk metals under UHV conditions. This work opens up a new approach to the study of surface diffusion of adsorbed molecules on nanoparticle electrode catalysts, including the possibility of correlating diffusion parameters to catalytic activity in real world applications of broad general interest. PMID- 16218594 TI - Stable metallic behavior and antiferromagnetic ordering of Fe(III) d spins in (EDO-TTFVO)2.FeCl4. AB - We report the crystal structure and physical properties of the 2:1 FeCl4- salt of a new donor molecule, EDO-TTFVO. Crystal structure analysis of this salt revealed that the donor molecules formed a beta' '-type two-dimensional conducting layer, and there is a short S...Cl contact between the donor molecules and the FeCl4- ions, which is expected to mediate a strong pi-d interaction. This salt showed a stable metallic conducting behavior down to 0.3 K and an antiferromagnetic ordering at TN approximately 3.0 K, indicating that this salt becomes a new antiferromagnetic molecular metal at ambient pressure. The appearance of the magnetic ordering is considered to originate from the strong pi-d interactions between the donor molecules and the FeCl4- ions because the field dependence of magnetoresistances was remarkably affected below the antiferromagnetic transition temperature. PMID- 16218595 TI - Self-assembly of programmed building blocks into structurally uniform dendrimers. AB - Selective and independent dimerization of tri- and tetraurea derivatives was used to build up dendritic assemblies which are uniform in size and structure. Dendrimers with the total molecular masses of about 25 000 g/mol were obtained. The existence of uniform assemblies was proved by 1H and 1H DOSY NMR experiments and also by dynamic light scattering. PMID- 16218596 TI - Transformation of isotropic fluid to nematic gel triggered by dynamic bridging of supramolecular nanocylinders. AB - We have synthesized an amphiphilic triblock coil-rod-coil molecule consisting of a rigid aromatic building block and poly(ethylene oxide). The coil-rod-coil molecule was observed to assemble into a cylindrical micelle structure in aqueous solution. Notably, addition of a small amount of a rod-coil-rod molecule into the cylindrical micellar solution can induce the anisotropic gelation due to dynamic interconnection of adjacent cylindrical micelles via hydrophobic and pi-pi interactions. Depending on the concentration of the rod-coil-rod molecule, the nematic gel can be reversibly switched into an isotropic solution of cylindrical micelles. PMID- 16218597 TI - CH activation with an O-donor iridium-methoxo complex. AB - A thermally and air stable O-donor, iridium-methoxo complex is reported that undergoes stoichiometric, intermolecular C-H activation of benzene with co generation of methanol and the iridium-phenyl complex. PMID- 16218598 TI - Evidence for the net addition of arene C-H bonds across a Ru(II)-hydroxide bond. AB - TpRu(PMe3)2(OH) (1) reacts with C6D6 to initiate H/D exchange between the hydroxide ligand and the deuterated benzene. In addition, complex 1 catalyzes H/D exchange between H2O and C6D6. Mechanistic and computational studies suggest that a likely reaction pathway for the H/D exchange involves loss of PMe3 to produce {TpRu(PMe3)(OH)}, followed by the net addition of a benzene C-H(D) bond across the Ru-OH bond to form the putative complex TpRu(PMe3)(OH2)(Ph). PMID- 16218599 TI - Persistence length control of the polyelectrolyte layer-by-layer self-assembly on carbon nanotubes. AB - We have studied layer-by-layer polyelectrolyte self-assembly on pristine individual single-wall carbon nanotubes as a function of solution ionic strength. We report the existence of an ionic strength threshold for the deposition, below which the majority of nanotubes remain uncoated. Once the ionic strength reaches the threshold value, the majority of the individual nanotubes become coated with polyelectrolytes. Our results indicate that the self-assembly process likely involves wrapping of polymer chains around nanotubes and that the polymer chain's ability to bend in order to accommodate the nanotube curvature is one of the critical parameters controlling layer-by-layer electrostatic self-assembly on these one-dimensional templates. PMID- 16218600 TI - Atomic layer deposition in nanometer-level replication of cellulosic substances and preparation of photocatalytic TiO2/cellulose composites. AB - TiO2 replicas of filter paper with nanometer-level accuracy were prepared by atomic layer deposition of thin conformal TiO2 coating, followed by a removal of the paper by air-anneal at 450 degrees C. Photocatalytic anatase TiO2/cellulose composites were also made by leaving the paper intact. The TiO2 films were deposited from Ti(OMe)4 and H2O at 150-250 degrees C. The photocatalytic activity of the TiO2/cellulose composite was verified by photocatalytic reduction of Ag(I) from an aqueous solution to Ag nanoparticles on the TiO2 surface. The TiO2/cellulose composites are mechanically more stable than the free-standing TiO2 replicas and are therefore potentially suitable as lightweight, high surface area photocatalysts. PMID- 16218602 TI - Biradical intermediate in the [2 + 2] photocycloaddition of dienes and alkenes to [60]fullerene. AB - The photocycloaddition of vinylcyclopropanes to C60 yields stereospecifically a five-membered [60]fullerene adduct. These results suggest a biradical intermediate of the [2 + 2] photocycloaddition between dienes or arylalkenes and C60. An electron transfer between the triplet excited state of C60 and the unsaturated substrates precedes the formation of the intermediate. PMID- 16218601 TI - Gold(III)-catalyzed nucleophilic substitution of propargylic alcohols. AB - Gold-catalyzed nucleophilic substitution on propargylic alcohols, with various C , O-, and S-nucleophiles, is described under very mild conditions (room temperature, dichloromethane) in 0-97% yield. PMID- 16218603 TI - Shape-controlled synthesis of manganese oxide octahedral molecular sieve three dimensional nanostructures. AB - Cryptomelane-type manganese oxide octahedral molecular (OMS) sieve three dimensional (3D) nanostructures were synthesized via facile temple-free low temperature hydrothermal reactions. Morphologies of the cryptomelane-type OMS-2 nanoparticles with tunnel dimension of 4.6 x 4.6 A can be tuned by varying reaction temperatures. At low temperature (120 degrees C), OMS-2 dendritic nanoclusters composed of uniform single-crystal nanotetragonal prisms with square cross-sections were formed. At high temperature (180 degrees C), the morphologies of OMS-2 became spherical dandelion-like microspheres composed of uniform single crystal OMS-2 nanoneedles. Slow oxidation of Mn(2+) by Cr(2)O(7)(-) under hydrothermal conditions is critical for the formation of the hierarchically ordered OMS-2 3D nanostructures. PMID- 16218604 TI - Dynamic kinetic asymmetric allylic alkylations of allenes. AB - The dynamic kinetic asymmetric allylic alkylations of racemic allene acetates has been developed with the DACH-phenyl Trost ligand 2 to give general access to allenes with high enantiomeric excess (84-95%) for both malonate and amine nucleophiles. Further, a most unusual dependence of enantioselectivity on base has been uncovered. The magnitude of the enantioselectivity is heavily dependent on the base for the malonate nucleophiles, but the sense and magnitude of the asymmetric induction is dependent on the base for the amine nucleophiles. A Rh(I) catalyzed intramolecular [4 + 2] cycloaddition of the DYKAT products was accomplished to afford formal Diels-Alder adducts, wherein the axial chirality is faithfully transferred into multiple stereogenic centers as well as olefin geometry. PMID- 16218606 TI - Dehydroaromatization of quinoxalines: one-step syntheses of trinuclear 1,6,7,12,13,18-hexaazatrinaphthylene titanium complexes. AB - We report the spontaneous coupling of N-heterocycles, initiated by C-H bond activation reactions. The reaction of quinoxalines and the titanocene acetylene complex Cp2Ti{eta2-C2(SiMe3)2}, as an excellent titanocene source, results in the formation of trinuclear 1,6,7,12,13,18-hexaazatrinaphthylene (HATN) titanium complexes. These HATN titanium complexes are thermally stable but sensitive to air and moisture. A three-fold dehydrogenative C-C coupling is proposed as the main step in the presented synthetic procedure. Particularly using commercial starting materials, an efficient route for the dehydrogenative coupling of N heterocycles, leading to multidentate ligands, has been established. PMID- 16218605 TI - Diazeniumdiolate ions as leaving groups in anomeric displacement reactions: a protection-deprotection strategy for ionic diazeniumdiolates. AB - Diazeniumdiolate ions [R2N-N(O)=N-O-] are of growing interest pharmacologically for their ability to generate up to two molar equivalents of bioactive nitric oxide (NO) spontaneously on protonating the amino nitrogen. Accordingly, their stability increases as the pH is raised. Here we show that the corresponding beta glucosides [R2N-N(O)=N-O-Glc] decreased in stability with pH; when R2N was diethylamino, the rate equation was kobs = ko + kOH- [OH-], where ko = 7.8 x 10-7 s-1 and kOH- = 5.3 x 10-3 M-1 s-1. The primary products were 1,6-anhydroglucose and the regenerated R2N-N(O)=N-O- ion. The results were qualitatively similar to those of beta-glucosyl fluoride and p-nitrophenoxide, whose hydrolyses have been rationalized as proceeding via a glycal oxide intermediate. This chemistry offers a convenient strategy for protecting heat- and acid-sensitive diazeniumdiolate ions during manipulations that would otherwise destroy them. As an example, a poly(urethane) film that generated NO in physiological buffer at a surface flux comparable to that of the mammalian vascular endothelium was prepared by glucosylating the ionic diazeniumdiolate group attached to the diol monomer before reacting it with the bis-isocyanate, then removing the saccharide with base when the protecting group was no longer needed. PMID- 16218607 TI - Alkylation of magnesium enamide with alkyl chlorides and fluorides. AB - A magnesium enamide derived from N-2-(N',N'-diethylamino)ethyl imine reacts with a primary and secondary alkyl chloride or a primary alkyl fluoride to give an alpha-alkylated ketone in good to excellent yield after hydrolysis. The reaction takes place with a high level of inversion of stereochemistry at the electrophilic carbon center and will be useful for production of optically active compounds. PMID- 16218608 TI - Simple alkenes as substitutes for organometallic reagents: nickel-catalyzed, intermolecular coupling of aldehydes, silyl triflates, and alpha olefins. AB - A nickel-catalyzed method for the three-component coupling of alkenes (ethylene and alpha olefins), aldehydes, and silyl triflates is described, and this process represents the first catalytic method for coupling aldehydes and alkenes to give allylic alcohol derivatives. Conceptually, the alkene functions as a replacement for an alkenylmetal reagent. PMID- 16218609 TI - Activation of C-H bonds of arenes: selectivity and reactivity in bis(pyridyl) platinum(II) complexes. AB - The reaction of [PtMe2(NN)] and B(C6F5)3/H2O in CF3CH2OH with arenes Ar-H gives [PtAr{HOB(C6F5)3}(LL)] if the bis(pyridyl) ligand NN forms a six-membered, but not five-membered, chelate ring; methyl-substituted arenes give selectivity for metalation of meta > para > ortho, but methoxy-substituted arenes give ortho > meta, para. PMID- 16218610 TI - N-tosyloxycarbamates as a source of metal nitrenes: rhodium-catalyzed C-H insertion and aziridination reactions. AB - The rhodium-catalyzed decomposition of N-tosyloxycarbamates to generate metal nitrenes which undergo intramolecular C-H insertion or aziridination reaction is described. Aliphatic N-tosyloxycarbamates produce oxazolidinones with high yields and stereospecificity through insertion in benzylic, tertiary, and secondary C-H bonds. Intramolecular aziridination occurs with allylic N-tosyloxycarbamates to produce aziridines as single diastereomers. The reaction proceeds at room temperature using a rhodium catalyst and an excess of potassium carbonate and does not require the use of strong oxidant, such as hypervalent iodine reagents. A rhodium nitrene species is presumably involved, as both reactions are stereospecific. PMID- 16218611 TI - Total synthesis of (+/-)-garsubellin A. AB - The first total synthesis of garsubellin A, a neurotrophic compound with potent choline acetyltransferase-inducing activity, is described. Keys for success were (1) stereoselective intermolecular aldol reaction at the C-4 position with acetaldehyde, (2) stereoelective Claisen rearrangement to introduce an allyl group to the most sterically crowded position at C-6, (3) ring-closing metathesis to construct the B-ring, and (4) Wacker-type oxidative C-ring formation. This synthetic route can be extended to an asymmetric synthesis of garsubellin A using the Koga catalytic enantioselective alkylation, which produced enantioenriched alpha-prenyl cyclohexenone with excellent enantioselectivity (95% ee). PMID- 16218612 TI - Unprecedented coordination modes and demetalation pathways for unbridged polyenyl ligands. Ruthenium eta1,eta4-cycloheptadienyl complexes from allyl/alkyne cycloaddition. AB - Cationic (eta6-hexamethylbenzene)ruthenium(II) mediates the [3 + 2 + 2] cycloaddition of allyl and alkyne ligands, leading to the unexpected isolation of eta1,eta4-cycloheptadienyl complexes, an unprecedented coordination mode for transition metal complexes of simple organic rings. The nonconjugated, eta1,eta4 coordinated complex is obtained as the kinetic reaction product from treatment of the unsubstituted allyl complex with excess ethyne; this complex rearranges slowly at 80 degrees C to the thermodynamically more stable conjugated eta5 cycloheptadienyl isomer. The eta1,eta4-coordinated isomer is fluxional at room temperature, undergoing rapid and reversible equilibration with a cycloheptatriene hydride intermediate via facile beta-hydride elimination/reinsertion. The reinsertion process is remarkably regioselective, returning the nonconjugated eta1,eta4-cycloheptadienyl isomer exclusively at room temperature. For reactions incorporating dimethylacetylene dicarboxylate (DMAD) as one or both of the alkyne components, eta1,eta4-coordination appears to be both kinetically and thermodynamically favored, despite undergoing equilibration among all possible eta1,eta4-cycloheptadienyl and cycloheptatriene hydride isomers prior to arriving at one observed eta1,eta4-isomer. For this series, no isomerization to eta5-coordination is observed even upon prolonged heating. In contrast, the cyclization incorporating both DMAD and phenylacetylene proceeds directly to the eta5-cycloheptadienyl isomer at or below room temperature, indicating that eta5-coordination remains energetically accessible to this system. The DMAD-based cyclization reactions produce structurally diverse minor byproducts, including both eta1,eta4-methanocyclohexadiene and acyclic eta3,eta2 heptadienyl isomers, which have been isolated and rigorously characterized. The unusual eta1,eta4-coordination of the seven-membered ring leads to unique new organic products upon oxidative demetalation by iodinolysis. Thus, reactions with excess iodine afford bridged tricyclic cyclopropane-containing lactones or substituted cycloheptatrienes in good but sometimes variable yields, depending on the substrate and specific reaction conditions. The ruthenium in these reactions is returned in high yield as the interesting cationic mu-triiodo pseudodimer of (eta6-hexamethylbenzene)ruthenium, which is obtained as a triiodide salt. This Ru(III) complex, along with several representative Ru(II) cyclization products, has been characterized in the solid state by X-ray crystallography. PMID- 16218613 TI - A mild radical procedure for the reduction of B-alkylcatecholboranes to alkanes. AB - A mild radical-mediated reduction of organoboranes is reported. The reducing agent is methanol complexed by the Lewis acidic B-methoxycatecholborane. PMID- 16218614 TI - Dichlorodiazirine: a nitrogenous precursor for dichlorocarbene. AB - The reaction of 3-p-nitrophenoxy-3-chlorodiazirine with a 1.1:1.0:1.6 blend of tetrabutylammonium chloride, cesium chloride, and the ionic liquid 1-butyl-3 methylimidazolium chloride at 40-50 degrees C under vacuum at 1 mm.Hg produced a melt from which dichlorodiazirine distilled into a cold trap containing pentane at -70 degrees C over 6-7 h. Dichlorodiazirine, the first nitrogenous precursor for dichlorocarbene, was reasonably stable in the dark at 25 degrees C and efficiently generated the carbene upon photolysis. PMID- 16218615 TI - Light-driven molecular motors: stepwise thermal helix inversion during unidirectional rotation of sterically overcrowded biphenanthrylidenes. AB - To investigate the unidirectional rotation of chiral overcrowded biphenanthrylidenes in more detail, the size of the substituent next to the double bond responsible for the unidirectionality of rotation was varied. The thermal and photochemical isomerization of three sterically overcrowded alkenes is described. The behavior of the biphenanthrylidenes with methyl and ethyl substituents is rather similar, and these compounds undergo a unidirectional 360 degrees rotation around the central double bond in a four-step sequence involving two photochemical cis-trans isomerizations and two thermal helix inversions. The only difference between these two true molecular motors was a small entropic effect, which causes the ethyl substituted molecular motor to rotate slightly faster. The behavior of the i-propyl substituted compound differs significantly from that of the other two. Although not all different isomers of the i-propyl substituted molecular motor were detected spectroscopically, experimental data led to the conclusion that this compound can also be considered as a molecular motor and is capable of performing a 360 degrees unidirectional rotation. (1)H NMR and X-ray analysis show a meso-like form as an intermediate in the unidirectional rotation, which proves that the thermal helix inversion is a stepwise process. PMID- 16218616 TI - Catalysis of enantioselective [2+1]-cycloaddition reactions of ethyl diazoacetate and terminal acetylenes using mixed-ligand complexes of the series Rh2(RCO2)n(L*4 n). Stereochemical heuristics for ligand exchange and catalyst synthesis. AB - This paper describes the synthesis of mixed Rh(2)(II) complexes containing bridging acetate and R,R-diphenyl-N-triflylimidazolidinone (DPTI) ligands (1, 2, and 9-19), and their function as enantioselective catalysts for the conversion of ethyl diazoacetate and terminal acetylenes to chiral cyclopropenes. Of these catalysts, 1 and 10 functioned with the highest enantioselectivity, in accord with a mechanistic model in which one of the ligand bridges is broken in the intermediate Rh-carbene complex. The synthetic results allow conclusions with regard to kinetically and thermodynamically favored pathways for the synthesis of mixed acetate-DPTI complexes. A new C(2)-symmetric complex having only two anti DTBTI bridges (23) is shown to be a highly effective chiral catalyst, as expected from the model. PMID- 16218617 TI - Structure and dynamics of solvated Sn(II) in aqueous solution: an ab initio QM/MM MD approach. AB - Structural and dynamical properties of the hydrated Sn(II) ion have been investigated by ab initio quantum mechanical/molecular mechanical (QM/MM) molecular dynamics (MD) simulations at double-zeta HF quantum mechanical level. The results indicate Sn(II)aq to be a rather peculiar, if not unique, case of a hydrated ion: four of its eight first-shell ligands do not take place in the otherwise frequent ligand-exchange processes, forming an approximately tetrahedral cage around the ion. The remaining ligands, however, exchange at a rate that is rather comparable to monovalent than divalent ions. This very surprising behavior of ligand exchange not yet observed in any previous simulation of over 30 hydrated metal ions is consistently confirmed by vibrational spectra, bond lengths, and a detailed analysis of the trajectories of the simulation. PMID- 16218619 TI - The coordination of uranyl in water: a combined quantum chemical and molecular simulation study. AB - The coordination environment of uranyl in water has been studied using a combined quantum mechanical and molecular dynamics approach. Multiconfigurational wave function calculations have been performed to generate pair potentials between uranyl and water. The quantum chemically determined energies have been used to fit parameters in a polarizable force field with an added charge transfer term. Molecular dynamics simulations have been performed for the uranyl ion and up to 400 water molecules. The results show a uranyl ion with five water molecules coordinated in the equatorial plane. The U-O(H(2)O) distance is 2.40 A, which is close to the experimental estimates. A second coordination shell starts at about 4.7 A from the uranium atom. No hydrogen bonding is found between the uranyl oxygens and water. Exchange of waters between the first and second solvation shell is found to occur through a path intermediate between association and interchange. This is the first fully ab initio determination of the solvation of the uranyl ion in water. PMID- 16218618 TI - Alkene cis-dihydroxylation by [(Me3tacn)(CF3CO2)RuVI O2)]ClO4(Me(3)tacn = 1,4,7 Trimethyl-1,4,7-triazacyclononane): structural characterization of [3 + 2] cycloadducts and kinetic studies. AB - cis-Dioxoruthenium(VI) complex [(Me(3)tacn)(CF(3)CO(2))Ru(VI)O(2)]ClO(4) (1, Me(3)tacn = 1,4,7-trimethyl-1,4,7-triazacyclononane) reacted with alkenes in aqueous tert-butyl alcohol to afford cis-1,2-diols in excellent yields under ambient conditions. When the reactions of 1 with alkenes were conducted in acetonitrile, oxidative C=C cleavage reaction prevailed giving carbonyl products in >90% yields without any cis-diol formation. The alkene cis-dihydroxylation and C=C cleavage reactions proceed via the formation of a [3 + 2] cycloadduct between 1 and alkenes, analogous to the related reactions with alkynes [Che et al. J. Am. Chem. Soc. 2000, 122, 11380]. With cyclooctene and trans-beta-methylstyrene as substrates, the Ru(III) cycloadducts (4a) and (4b) [formula; see text] were isolated and structurally characterized by X-ray crystal analyses. The kinetics of the reactions of 1 with a series of p-substituted styrenes has been studied in acetonitrile by stopped-flow spectrophotometry. The second-order rate constants varied by 14-fold despite an overall span of 1.3 V for the one-electron oxidation potentials of alkenes. Secondary kinetic isotope effect (KIE) was observed for the oxidation of beta-d(2)-styrene (k(H)/k(D) = 0.83 +/- 0.04) and alpha deuteriostyrene (k(H)/k(D) = 0.96 +/- 0.03), which, together with the stereoselectivity of cis-alkene oxidation by 1, is in favor of a concerted mechanism. PMID- 16218620 TI - Charge recombination via intercolumnar electron tunneling through the lipid-like mantle of discotic hexa-alkyl-hexa-peri-hexabenzocoronenes. AB - The recombination of the mobile charge carriers formed in pulse-ionized hexa alkyl-substituted hexa-peri-hexabenzocoronenes occurs mainly via intercolumnar electron tunneling through the intervening hydrocarbon mantle. This is evidenced as a dramatic increase in the time scale of the decay of the radiation-induced conductivity from a few hundred nanoseconds to close to a millisecond as the peripheral alkyl substituents increase in size from 8 to 24 carbon atoms with corresponding disk diameters, D, from 23.4 to 36.6 A. The decay kinetics are a function only of the total number of peripheral carbon atoms with no evidence for specific effects of chain branching. The 1/e decay time, tau(e), increases exponentially with D according to tau(e) = tau(e)(0) exp(betaD) with tau(e)(0) = 48 fs and beta = 0.63 A(-1). Taking into account the tilted columnar configuration of the molecules in the solid phase leads to a beta value of ca. 0.8 A(-1) for the distance dependence of intercolumnar electron tunneling. In contrast to the orders of magnitude changes in the time scale for intercolumnar charge recombination, the intracolumnar charge hopping times vary by only a factor of 4, between 40 and 160 fs, with no systematic dependence on the nature of the alkyl substituents. On the basis of the results, the time scale estimated for electron tunneling across a 40 A thick lipid membrane is estimated to be close to 1 ms. PMID- 16218621 TI - Mechanism of the mild functionalization of arenes by diboron reagents catalyzed by iridium complexes. Intermediacy and chemistry of bipyridine-ligated iridium trisboryl complexes. AB - This paper describes mechanistic studies on the functionalization of arenes with the diboron reagent B(2)pin(2) (bis-pinacolato diborane(4)) catalyzed by the combination of 4,4'-di-tert-butylbipyridine (dtbpy) and olefin-ligated iridium halide or olefin-ligated iridium alkoxide complexes. This work identifies the catalyst resting state as [Ir(dtbpy)(COE)(Bpin)(3)] (COE = cyclooctene, Bpin = 4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl). [Ir(dtbpy)(COE)(Bpin)(3)] was prepared by independent synthesis in high yield from [Ir(COD)(OMe)](2), dtbpy, COE, and HBpin. This complex is formed in low yield from [Ir(COD)(OMe)](2), dtbpy, COE, and B(2)pin(2). Kinetic studies show that this complex reacts with arenes after reversible dissociation of COE. An alternative mechanism in which the arene reacts with the Ir(I) complex [Ir(dtbpy)Bpin] after dissociation of COE and reductive elimination of B(2)pin(2) does not occur to a measurable extent. The reaction of [Ir(dtbpy)(COE)(Bpin)(3)] with arenes and the catalytic reaction of B(2)pin(2) with arenes catalyzed by [Ir(COD)(OMe)](2) and dtbpy occur faster with electron-poor arenes than with electron-rich arenes. However, both the stoichiometric and catalytic reactions also occur faster with the electron-rich heteroarenes thiophene and furan than with arenes, perhaps because eta(2) heteroarene complexes are more stable than the eta(2)-arene complexes and the eta(2)-heteroarene or arene complexes are intermediates that precede oxidative addition. Kinetic studies on the catalytic reaction show that [Ir(dtbpy)(COE)(Bpin)(3)] enters the catalytic cycle by dissociation of COE, and a comparison of the kinetic isotope effects of the catalytic and stoichiometric reactions shows that the reactive intermediate [Ir(dtbpy)(Bpin)(3)] cleaves the arene C-H bond. The barriers for ligand exchange and C-H activation allow an experimental assessment of several conclusions drawn from computational work. Most generally, our results corroborate the conclusion that C-H bond cleavage is turnover-limiting, but the experimental barrier for this bond cleavage is much lower than the calculated barrier. PMID- 16218622 TI - The influence of aqueous versus glassy solvents on protein dynamics: vibrational echo experiments and molecular dynamics simulations. AB - Spectrally resolved infrared stimulated vibrational echo measurements are used to measure the vibrational dephasing of the CO stretching mode of carbonmonoxy hemoglobin (HbCO), a myoglobin mutant (H64V), and a bacterial cytochrome c(552) mutant (Ht-M61A) in aqueous solution and trehalose glasses. The vibrational dephasing of the heme-bound CO is significantly slower for all three proteins embedded in trehalose glasses compared to that of aqueous protein solutions. All three proteins exhibit persistent but notably slower spectral diffusion when the protein surface is fixed by the glassy solvent. Frequency-frequency correlation functions (FFCFs) of the CO are extracted from the vibrational echo data to reveal that the structural dynamics, as sensed by the CO, of the three proteins in trehalose and aqueous solution are dominated by fast (tens of femtoseconds), motionally narrowed fluctuations. MD simulations of H64V in dynamic and "static" water are presented as models of the aqueous and glassy environments. FFCFs are calculated from the H64V simulations and qualitatively reproduce the important features of the experimentally extracted FFCFs. The suppression of long time scale (picoseconds to tens of picoseconds) frequency fluctuations (spectral diffusion) in the glassy solvent is the result of a damping of atomic displacements throughout the protein structure and is not limited to structural dynamics that occur only at the protein surface. The analysis provides evidence that some dynamics are coupled to the hydration shell of water, supporting the idea that the bioprotection offered by trehalose is due to its ability to immobilize the protein surface through a thin, constrained layer of water. PMID- 16218623 TI - Magnetic field dependence of photo-CIDNP MAS NMR on photosynthetic reaction centers of Rhodobacter sphaeroides WT. AB - Photochemically induced dynamic nuclear polarization (photo-CIDNP) is observed in frozen and quinone depleted photosynthetic reaction centers of the purple bacteria Rhodobacter sphaeroides wild type (WT) by (13)C solid-state NMR at three different magnetic fields. All light-induced signals appear to be emissive at all three fields. At 4.7 T (200 MHz proton frequency), the strongest enhancement of NMR signals is observed, which is more than 10 000 above the Boltzmann polarization. At higher fields, the enhancement factor decreases. At 17.6 T, the enhancement factor is about 60. The field dependence of the enhancement appears to be the same for all nuclei. The observed field dependence is in line with simulations that assume two competing mechanisms of polarization transfer from electrons to nuclei, three-spin mixing (TSM) and differential decay (DD). These simulations indicate a ratio of the electron spin density on the special pair cofactors is 3:2 in favor of the L-BChl during the radical cation state. The good agreement of simulations with the experiments raises expectations that artificial solid reaction centers can be tuned to show photo-CIDNP in the near future. PMID- 16218624 TI - Covalent attachment of a transition metal coordination complex to functionalized oligo(phenylene-ethynylene) self-assembled monolayers. AB - We have investigated the reaction of tetrakis(dimethylamido)titanium, Ti[N(CH(3))(2)](4), with N-isopropyl-N-[4-(thien-3-ylethynyl) phenyl] amine and N isopropyl-N-(4-{[4-(thien-3-ylethynyl) phenyl]ethynyl}phenyl) amine self assembled monolayers (SAMs), on polycrystalline Au substrates. The structure of the SAMs themselves has also been investigated. Both molecules form SAMs on polycrystalline Au bound by the thiophene group. The longer-molecular-backbone molecule forms a denser SAM, with molecules characterized by a smaller tilt angle. X-ray photoelectron spectroscopy (XPS) and angle-resolved XPS have been employed to examine the kinetics of adsorption, the spatial extent of reaction, and the stoichiometry of reaction. For both the SAMs, adsorption is described well by first-order Langmuirian kinetics, and adsorption is self-limiting from T(s) = -50 to 30 degrees C. The use of angle-resolved XPS clearly demonstrates that the Ti[N(CH(3))(2)](4) reacts exclusively with the isopropylamine end group via ligand exchange, and there is no penetration of the SAM, followed by reaction at the SAM-Au interface. Moreover, the SAM molecules remain bound to the Au surface via their thiopene functionalites. From XPS, we have found that, in both cases, approximately one Ti[N(CH(3))(2)](4) is adsorbed per two SAM molecules. PMID- 16218625 TI - Naked (C5Me5)2M cations (M = Sc, Ti, and V) and their fluoroarene complexes. AB - The ionic metallocene complexes [Cp*(2)M][BPh(4)] (Cp* = C(5)Me(5)) of the trivalent 3d metals Sc, Ti, and V were synthesized and structurally characterized. For M = Sc, the anion interacts weakly with the metal center through one of the phenyl groups, but for M = Ti and V, the cations are naked. They each contain one strongly distorted Cp* ligand, with one (V) or two (Ti) agostic C-H...M interactions involving the Cp*Me groups. For Sc and Ti, these Lewis acidic species react with fluorobenzene and 1,2-difluorobenzene to yield [Cp*(2)M(kappaF-FC(6)H(5))(n)][BPh(4)] (M = Sc, n = 2; M = Ti, n = 1) and [Cp*(2)M(kappa(2)F-1,2-F(2)C(6)H(4))][BPh(4)], the first examples of kappaF fluorobenzene and kappa(2)F-1,2-difluorobenzene adducts of transition metals. With the perfluorinated anion [B(C(6)F(5))(4)](-), both Sc and Ti form [Cp*(2)M(kappa(2)F-C(6)F(5))B(C(6)F(5))(3)] contact ion pairs. The nature of the metal-fluoroarene interaction was studied by density functional theory (DFT) calculations and by comparison with the corresponding tetrahydrofuran (THF) adducts and was found to be predominantly electrostatic for all metals studied. PMID- 16218626 TI - Interpreting NMR data for beta-peptides using molecular dynamics simulations. AB - NMR is one of the most used techniques to resolve structure of proteins and peptides in solution. However, inconsistencies may occur due to the fact that a polypeptide may adopt more than one conformation. Since the NOE distance bounds and (3)J-values used in such structure determination represent a nonlinear average over the total ensemble of conformers, imposition of NOE or (3)J-value restraints to obtain one unique conformation is not an appropriate procedure in such cases. Here, we show that unrestrained MD simulation of a solute in solution using a high-quality force field yields a conformational ensemble that is largely compatible with the experimental NMR data on the solute. Four 100 ns MD simulations of two forms of a nine-residue beta-peptide in methanol at two temperatures produced conformational ensembles that were used to interpret the NMR data on this molecule and resolve inconsistencies between the experimental NOEs. The protected and unprotected forms of the beta-peptide adopt predominantly a 12/10-helix in agreement with the qualitative interpretation of the NMR data. However, a particular NOE was not compatible with this helix indicating the presence of other conformations. The simulations showed that 3(14)()-helical structures were present in the ensemble of the unprotected form and that their presence correlates with the fulfillment of the particular NOE. Additionally, all inter-hydrogen distances were calculated to compare NOEs predicted by the simulations to the ones observed experimentally. The MD conformational ensembles allowed for a detailed and consistent interpretation of the experimental data and showed the small but specific conformational differences between the protected and unprotected forms of the peptide. PMID- 16218627 TI - Theoretical studies on the magnetic switching controlled by stacking patterns of bis(maleonitriledithiolato) nickelate(III) dimers. AB - Magnetic switchable maleonitriledithiolate (mnt) complexes were studied by density functional theory. The calculations were performed for anion dimers of [RBzPyR'][Ni(mnt)(2)] (RBzPyR' = derivatives of benzylpyridinium) to elucidate magnetostructural correlations and the nature of the weak intermolecular chemical bonding. The calculated results showed that the spin delocalization, favored by the eclipsed stacking and the shorter interlayer distance, was responsible for the diamagnetic character of [1-benzyl-4-aminopyridinium][Ni(mnt)(2)] at low temperature. The weak antiferromagnetic and ferromagnetic interactions were also reproduced for [1-benzyl-4-aminopyridinium][Ni(mnt)(2)] and [1-(4' fluorobenzyl)pyridinium][Ni(mnt)(2)] at high temperature, respectively. The natural bond orbital analysis suggested that the cooperative effect of the weak intermolecular bondings may be the intrinsic driving force resulting in the switchable property, which is essentially similar to those in organic radicals exhibiting magnetic bistability. Further investigations with varying interlayer distance d, the extent of slippage (slipping distance r and deviation angle alpha), and rotational angle theta suggested that the extent of slippage played an important role in magnetic interactions. Therefore, the abrupt modulation of the extent of slippage in the [Ni(mnt)(2)](-) complexes by external perturbations provided new possibilities for the design of molecular magnetic switching devices. PMID- 16218628 TI - Catalytic mechanism of limonene epoxide hydrolase, a theoretical study. AB - The catalytic mechanism of limonene epoxide hydrolase (LEH) was investigated theoretically using the density functional theory method B3LYP. LEH is part of a novel limonene degradation pathway found in Rhodococcus erythropolis DCL14, where it catalyzes the hydrolysis of limonene-1,2-epoxide to give limonene-1,2-diol. The recent crystal structure of LEH was used to build a model of the LEH active site composed of five amino acids and a crystallographically observed water molecule. With this model, hydrolysis of different substrates was investigated. It is concluded that LEH employs a concerted general acid/general base-catalyzed reaction mechanism involving protonation of the substrate by Asp101, nucleophilic attack by water on the epoxide, and abstraction of a proton from water by Asp132. Furthermore, we provide an explanation for the experimentally observed regioselective hydrolysis of the four stereoisomers of limonene-1,2-epoxide. PMID- 16218629 TI - Hydrogen cycling of niobium and vanadium catalyzed nanostructured magnesium. AB - The reaction of hydrogen gas with magnesium metal, which is important for hydrogen storage purposes, is enhanced significantly by the addition of catalysts such as Nb and V and by using nanostructured powders. In situ neutron diffraction on MgNb(0.05) and MgV(0.05) powders give a detailed insight on the magnesium and catalyst phases that exist during the various stages of hydrogen cycling. During the early stage of hydriding (and deuteriding), a MgH(1< x < 2) phase is observed, which does not occur in bulk MgH(2) and, thus, appears characteristic for the small particles. The abundant H vacancies will cause this phase to have a much larger hydrogen diffusion coefficient, partly explaining the enhanced kinetics of nanostructured magnesium. It is shown that under relevant experimental conditions, the niobium catalyst is present as NbH(1). Second, a hitherto unknown Mg-Nb perovskite phase could be identified that has to result from mechanical alloying of Nb and the MgO layer of the particles. Vanadium is not visible in the diffraction patterns, but electron micrographs show that the V particle size becomes very small, 2-20 nm. Nanostructuring and catalyzing the Mg enhance the adsorption speed that much that now temperature variations effectively limit the absorption speed and not, as for bulk, the slow kinetics through bulk MgH(2) layers. PMID- 16218630 TI - Interstrand and intrastrand DNA-DNA cross-linking by 1,2,3,4-diepoxybutane: role of stereochemistry. AB - 1,2,3,4-Diepoxybutane (DEB) is a bifunctional electrophile capable of forming DNA DNA and DNA-protein cross-links. DNA alkylation by DEB produces N7-(2'-hydroxy 3',4'-epoxybut-1'-yl)-guanine monoadducts, which can then form 1,4-bis-(guan-7 yl)-2,3-butanediol (bis-N7G-BD) lesions. All three optical isomers of DEB are produced metabolically from 1,3-butadiene, but S,S-DEB is the most cytotoxic and genotoxic. In the present work, interstrand and intrastrand DNA-DNA cross-linking by individual DEB stereoisomers was investigated by PAGE, mass spectrometry, and stable isotope labeling. S,S-, R,R-, and meso-diepoxides were synthesized from l dimethyl-2,3-O-isopropylidene-tartrate, d-dimethyl-2,3-O-isopropylidene-tartrate, and meso-erythritol, respectively. Total numbers of bis-N7G-BD lesions (intrastrand and interstrand) in calf thymus DNA treated separately with S,S-, R,R-, or meso-DEB (0.01-0.5 mM) were similar as determined by capillary HPLC ESI(+)-MS/MS of DNA hydrolysates. However, denaturing PAGE has revealed that S,S DEB produced the highest number of interchain cross-links in 5'-GGC-3'/3'-CCG-5' sequences. Intrastrand adduct formation by DEB was investigated by a novel methodology based on stable isotope labeling HPLC-ESI(+)-MS/MS. Meso DEB treatment of DNA duplexes containing 5'-[1,7, NH(2)-(15)N(3),2-(13)C-G]GC-3'/3' CCG-5' and 5'-GGC-3'/3'-CC[(15)N(3),2-(13)C-G]-5' trinucleotides gave rise to comparable numbers of 1,2-intrastrand and 1,3-interstrand bis-N7G-BD cross-links, while S,S DEB produced few intrastrand lesions. R,R-DEB treated DNA contained mostly 1,3-interstrand bis-N7G-BD, along with smaller amounts of 1,2-interstrand and 1,2-intrastrand adducts. The effects of DEB stereochemistry on its ability to form DNA-DNA cross-links may be rationalized by the spatial relationships between the epoxy alcohol side chains in stereoisomeric N7-(2'-hydroxy-3',4'-epoxybut-1' yl)-guanine adducts and their DNA environment. Different cross-linking specificities of DEB stereoisomers provide a likely structural basis for their distinct biological activities. PMID- 16218631 TI - Synthesis and crystal structure of an open capsule-type octanuclear heterometallic sulfide cluster with a linked incomplete double cubane framework without an intramolecular inversion center. AB - An open capsule-type octanuclear heterometallic sulfide cluster without an intramolecular inversion center [Ru(eta(6)-C(6)Me(6)){P(OMe)(3)}{MoO(mu(3) S)(3)}(CuI)(2)](2) (5) has been synthesized for the first time by stepwise connection of three mononuclear building blocks, i.e., (i) [RuCl(2)(eta(6) C(6)Me(6)){P(OMe)(3)}] (1a) as an octahedral terminal building block to control the direction of cluster expansion, (ii) [MoOS(3)](2)(-) as a tetrahedral polydentate building block owing to the strong coordination ability of the S atoms, and (iii) a CuI building block to form a trigonal planar (mu-S)(2)CuI unit or to form a linkage unit of two incomplete cubane-type octanuclear frameworks. The stepwise connection was made in the following order: [RuCl(2)(eta(6) C(6)Me(6)){P(OMe)(3)}] (1a, mononuclear) --> [Ru(eta(6) C(6)Me(6)){P(OMe)(3)}{MoOS(mu(2)-S)(2)}] (2a, dinuclear) --> [Ru(eta(6) C(6)Me(6)){P(OMe)(3)}{MoO(mu(2)-S)(2)(mu(3)-S)}CuI] (3a, butterfly-type trinuclear) --> [Ru(eta(6)-C(6)Me(6)){P(OMe)(3)}{MoO(mu(3)-S)(3)}(CuI)(2)](2) (5). When P(OMe)(3) was replaced by P(OEt)(3), which is more bulky than P(OMe)(3), in the starting ruthenium building block [RuCl(2)(eta(6) C(6)Me(6)){P(OEt)(3)}] (1b, mononuclear), only the tetranuclear incomplete single cubane cluster [Ru(eta(6)-C(6)Me(6)){P(OEt)(3)}{MoO(mu(3)-S)(3)}(CuI)(2)] (6) was generated, owing to the steric effect of P(OEt)(3). PMID- 16218632 TI - Diastereomer-differentiating photochemistry of beta-arylbutyrophenones: Yang cyclization versus type II elimination. AB - The diastereomers of ketones 2 and 3 are shown to exhibit distinct photochemical reactivities due to conformational preferences; while the anti isomers of 2 and 3 undergo efficient Yang cyclization in 75-90% yields with a remarkable diastereoselectivity (> 90%), the syn isomers predominantly undergo Norrish Type II elimination. The differences in the product profiles of the diastereomers are consistent with a mechanistic picture involving the formation of precursor diastereomeric triplet 1,4-biradicals in which the substituents at alpha and beta positions stabilize the cisoid (cyclization) or transoid (elimination) geometry. The fact that such a diastereomeric relationship does indeed ensue at the triplet excited-state itself is demonstrated via the nanosecond laser-flash photolysis of model ketones 1. The diastereomeric discrimination in the product profiles observed for ketones 2 and 3 as well as in the triplet lifetimes observed for ketones 1 can both be mechanistically traced back to different conformational preferences of the ground-state diastereomeric ketones and the intermediary 1,4 biradicals. Additionally, it emerges from the present study that the syn and anti diastereomers of ketones 2 and 3 represent two extremes of a broad range of widely examined butyrophenones, which lead to varying degrees of Yang photocyclization depending on the alkyl substitution pattern. PMID- 16218633 TI - Synthesis of lipidated green fluorescent protein and its incorporation in supported lipid bilayers. AB - Herein we report a semisynthetic method of producing membrane-anchored proteins. Ligation of synthetic lipids with designed anchor structures to proteins was performed using native chemical ligation (NCL) of a C-terminal peptide thioester and an N-terminal cysteine lipid. This strategy mimics the natural glycosylphosphatidylinositol (GPI) linkage found in many natural membrane associated proteins; however, the synthetic method utilizes simple lipid anchors without glycans. Synthetically lipidated recombinant green fluorescent protein (GFP) was shown to be stably anchored to the membrane, and its lateral fluidity was quantitatively characterized by direct fluorescence imaging in supported membranes. Circumventing the steps of purification from native cell membranes, this methodology facilitates the reconstitution of membrane-associated proteins. PMID- 16218634 TI - Modeling spontaneous formation of precursor nanoparticles in clear-solution zeolite synthesis. AB - We present a lattice model describing the formation of silica nanoparticles in the early stages of the clear-solution templated synthesis of silicalite-1 zeolite. Silica condensation/hydrolysis is modeled by a nearest-neighbor attraction, while the electrostatics are represented by an orientation-dependent, short-range interaction. Using this simplified model, we show excellent qualitative agreement with published experimental observations. The nanoparticles are identified as a metastable state, stabilized by electrostatic interactions between the negatively charged silica surface and a layer of organic cations. Nanoparticle size is controlled mainly by the solution pH, through nanoparticle surface charge. The size and concentration of the charge-balancing cation are found to have a negligible effect on nanoparticle size. Increasing the temperature allows for further particle growth by Ostwald ripening. We suggest that this mechanism may play a role in the growth of zeolite crystals. PMID- 16218636 TI - A molecular tweezer for lysine and arginine. AB - Lysine and arginine play a key role in numerous biological recognition processes controlling, inter alia, gene regulation, glycoprotein targeting and vesicle transport. They are also found in signaling peptide sequences responsible, e.g. for bacterial cell wall biosynthesis, Alzheimer peptide aggregation and skin regeneration. Almost none of all artificial receptor structures reported to date are selective and efficient for lysine residues in peptides or proteins. An artificial molecular tweezer is introduced which displays an exceptionally high affinity for lysine (K(a) approximately 5000 in neutral phosphate buffer). It features an electron-rich torus-shaped cavity adorned with two peripheral anionic phosphonate groups. Exquisite selectivity for arginine and lysine is achieved by threading the whole amino acid side chain through the cavity and subsequent locking by formation of a phosphonate-ammonium/guanidinium salt bridge. This pseudorotaxane-like geometry is also formed in small basic signaling peptides, which can be bound with unprecedented affinity in buffered aqueous solution. NMR titrations, NOESY and VT experiments as well as ITC measurements and Monte Carlo simulations unanimously point to an enthalpy-driven process utilizing a combination of van der Waals interactions and substantial electrostatic contributions for a conformational lock. Since DMSO and acetonitrile compete with the amino acid guest inside the cavity, a simple change in the cosolvent composition renders the whole complexation process reversible. PMID- 16218635 TI - Modeling multiple species of nicotine and deschloroepibatidine interacting with alpha4beta2 nicotinic acetylcholine receptor: from microscopic binding to phenomenological binding affinity. AB - A variety of molecular modeling, molecular docking, and first-principles electronic structure calculations were performed to study how the alpha4beta2 nicotinic acetylcholine receptor (nAChR) binds with different species of two typical agonists, (S)-(-)-nicotine and (R)-(-)-deschloroepibatidine, each of which is distinguished by different free bases and protonation states. On the basis of these results, predictions were made regarding the corresponding microscopic binding free energies. Hydrogen-bonding and cation-pi interactions between the receptor and the respective ligands were found to be the dominant factors differentiating the binding strengths of different microscopic binding species. The calculated results and analyses demonstrate that, for each agonist, all the species are interchangeable and can quickly achieve a thermodynamic equilibrium in solution and at the nAChR binding site. This allows quantitation of the equilibrium concentration distributions of the free ligand species and the corresponding microscopic ligand-receptor binding species, their pH dependence, and their contributions to the phenomenological binding affinity. The predicted equilibrium concentration distributions, pK(a) values, absolute phenomenological binding affinities, and their pH dependence are all in good agreement with available experimental data, suggesting that the computational strategy from the microscopic binding species and affinities to the phenomenological binding affinity is reliable for studying alpha4beta2 nAChR-ligand binding. This should provide valuable information for future rational design of drugs targeting nAChRs. The general strategy of the "from-microscopic-to-phenomenological" approach for studying interactions of alpha4beta2 nAChRs with (S)-(-)-nicotine and (R)-(-)-deschloroepibatidine may also be useful in studying other types of ligand-protein interactions involving multiple molecular species of a ligand and in associated rational drug design. PMID- 16218637 TI - Heme carbonyls: environmental effects on nu(C-O) and Fe-C/C-O bond length correlations. AB - The synthesis and characterization of four low-spin (carbonyl)iron(II) tetraphenylporphyrinates, [Fe(TPP)(CO)(L)], where L = 1-methylimidazole, 2 methylimidazole, 1,2-dimethylimidazole (unsolvated), and 1,2-dimethylimidazole (toluene solvate) are reported. The complexes show nearly the same value of nu(C O) in toluene solution (1969-72 cm(-1)) but a large range of CO stretching frequencies in the solid-state (1926-1968 cm(-1)). The large solid-state variation results from CO interactions in the solid state, as shown by an examination of the crystal structures of the four complexes. The high precision of the four structures obtained allows us to make a number of structural and spectroscopic correlations that describe the Fe-C-O and N(Im)-Fe-CO units. The values of nu(C-O) and the Fe-C and C-O bond distances are strongly correlated and provide a structural, as well as a spectroscopic, correlation of the pi back bonding model. The interactions of CO described are closely related to the large range of CO stretching frequencies observed in heme proteins and specific interactions observed in carbonylmyoglobin (MbCO). PMID- 16218638 TI - Detection and automatic repair of nucleotide base-pair mutations by coherent light. AB - We show that phase-coherent optical techniques allow for the detection and automatic repair of mutations in nucleotide pairs. We demonstrate computationally that there is a laser pulse sequence that can detect the occurrence of a mutation caused by a double proton transfer between hydrogen-bonded nucleotide pairs and automatically repair it by converting the mutated nucleotide pair to the nonmutated one. The specific system chosen for this demonstration is the hydrogen bonded 2-pyridone.2-hydroxypyridine dimer at typical internucleotide distances, a well-established model for tautomeric acid base pairs. PMID- 16218639 TI - Antibiotic recognition by binuclear metallo-beta-lactamases revealed by X-ray crystallography. AB - Metallo-beta-lactamases are zinc-dependent enzymes responsible for resistance to beta-lactam antibiotics in a variety of host bacteria, usually Gram-negative species that act as opportunist pathogens. They hydrolyze all classes of beta lactam antibiotics, including carbapenems, and escape the action of available beta-lactamase inhibitors. Efforts to develop effective inhibitors have been hampered by the lack of structural information regarding how these enzymes recognize and turn over beta-lactam substrates. We report here the crystal structure of the Stenotrophomonas maltophilia L1 enzyme in complex with the hydrolysis product of the 7alpha-methoxyoxacephem, moxalactam. The on-enzyme complex is a 3'-exo-methylene species generated by elimination of the 1 methyltetrazolyl-5-thiolate anion from the 3'-methyl group. Moxalactam binding to L1 involves direct interaction of the two active site zinc ions with the beta lactam amide and C4 carboxylate, groups that are common to all beta-lactam substrates. The 7beta-[(4-hydroxyphenyl)malonyl]-amino substituent makes limited hydrophobic and hydrogen bonding contacts with the active site groove. The mode of binding provides strong evidence that a water molecule situated between the two metal ions is the most likely nucleophile in the hydrolytic reaction. These data suggest a reaction mechanism for metallo-beta-lactamases in which both metal ions contribute to catalysis by activating the bridging water/hydroxide nucleophile, polarizing the substrate amide bond for attack and stabilizing anionic nitrogen intermediates. The structure illustrates how a binuclear zinc site confers upon metallo-beta-lactamases the ability both to recognize and efficiently hydrolyze a wide variety of beta-lactam substrates. PMID- 16218640 TI - DNA as helical ruler: exciton-coupled circular dichroism in DNA conjugates. AB - The structure and properties of oligonucleotide conjugates possessing stilbenedicarboxamide chromophores at both ends of a poly(dA):poly(dT) base-pair domain of variable length have been investigated using a combination of spectroscopic and computational methods. These conjugates form capped hairpin structures in which one stilbene serves as a hairpin linker and the other as a hydrophobic end-cap. The capping stilbene stabilizes the hairpin structures by ca. 2 kcal/mol, making possible the formation of a stable folded structure containing a single A:T base pair. Exciton coupling between the stilbene chromophores has little effect on the absorption bands of capped hairpins. However, exciton-coupled circular dichroism (EC-CD) can be observed for capped hairpins possessing as many as 11 base pairs. Both the sign and intensity of the EC-CD spectrum are sensitive to the number of base pairs separating the stilbene chromophores, as a consequence of the distance and angular dependence of exciton coupling. Calculated spectra obtained using a static vector model based on canonical B-DNA are in good agreement with the experimental spectra. Molecular dynamics simulations show that conformational fluctuations of the capped hairpins result in large deviations of the averaged spectra in both the positive and negative directions. These results demonstrate for the first time the ability of B-DNA to serve as a helical ruler for the study of electronic interactions between aligned chromophores. Furthermore, they provide important tests for atomistic theoretical models of DNA. PMID- 16218641 TI - Unraveling the crystallization mechanism of CoAPO-5 molecular sieves under hydrothermal conditions. AB - The hydrothermal crystallization of CoAPO-5 molecular sieves has been studied using time-resolved in-situ SAXS/WAXS, UV-vis, Raman, and XAS. Data collected during heating to 180 degrees C allowed the observation of different steps occurring during the transformation of the amorphous gel into a crystalline material from a macroscopic and atomic perspective. Raman spectroscopy detected the initial formation of Al-O-P bonds, whereas SAXS showed that these gel particles had a broad size distribution ranging from ca. 7 to 20 nm before crystallization began. WAXS showed that this crystallization was sharp and occurred at around 160 degrees C. Analysis of the crystallization kinetics suggested a one-dimensional growth process. XAS showed that Co(2+) transformed via a two-stage process during heating involving (i) a gradual transformation of octahedral coordination into tetrahedral coordination before the appearance of Bragg peaks corresponding to AFI, suggesting progressive incorporation of Co(2+) into the poorly ordered Al-O-P network up to ca. 150 degrees C, and (ii) a rapid transformation of remaining octahedral Co(2+) at the onset of crystallization. Co(2+) was observed to retard crystallization of AFI but provided valuable information regarding the synthesis process by acting as an internal probe. A three-stage, one-dimensional crystallization mechanism is proposed: (i) an initial reaction between aluminum and phosphate units forming a primary amorphous phase, (ii) progressive condensation of linear Al-O-P chains forming a poorly ordered structure separated by template molecules up to ca. 155 degrees C, and (iii) rapid internal reorganization of the aluminophosphate network leading to crystallization of the AFI crystal structure. PMID- 16218642 TI - Characterization of the cation and temperature dependence of DNA quadruplex hydrogen bond properties using high-resolution NMR. AB - Variations in the hydrogen bond network of the Oxy-1.5 DNA guanine quadruplex have been monitored by trans-H-bond scalar couplings, (h2)J(N2N7), for Na(+)-, K(+)-, and NH(4)(+)-bound forms over a temperature range from 5 to 55 degrees C. The variations in (h2)J(N2N7) couplings exhibit an overall trend of Na(+) > K(+) > NH(4)(+) and correlate with the different cation positions and N2-H2...N7 H bond lengths in the respective structures. A global weakening of the (h2)J(N2N7) couplings with increasing temperature for the three DNA quadruplex species is accompanied by a global increase of the acceptor (15)N7 chemical shifts. Above 35 degrees C, spectral heterogeneity indicates thermal denaturation for the Na(+) bound form, whereas spectral homogeneity persists up to 55 degrees C for the K(+) and NH(4)(+)-coordinated forms. The average relative change of the (h2)J(N2N7) couplings amounts to approximately 0.8 x 10(-3)/K and is thus considerably smaller than respective values reported for nucleic acid duplexes. The significantly higher thermal stability of H-bond geometries in the DNA quadruplexes can be rationalized by their cation coordination of the G-quartets and the extensive H-bond network between the four strands. A detailed analysis of individual (h2)J(N2N7) couplings reveals that the 5' strand end, comprising base pairs G1-G9* and G4*-G1, is the most thermolabile region of the DNA quadruplex in all three cation-bound forms. PMID- 16218643 TI - Strong resistance of phosphorylcholine self-assembled monolayers to protein adsorption: insights into nonfouling properties of zwitterionic materials. AB - In this work, we show the strong resistance of zwitterionic phosphorylcholine (PC) self-assembled monolayers (SAMs) to protein adsorption and examine key factors leading to their nonfouling behavior using both experimental and molecular simulation techniques. Zwitterions with a balanced charge and minimized dipole are excellent candidates as nonfouling materials due to their strong hydration capacity via electrostatic interactions. PMID- 16218645 TI - Small-molecule dynamics and mechanisms underlying the macroscopic mechanical properties of coordinatively cross-linked polymer networks. AB - Specific metal-ligand coordination between bis-Pd(II) and Pt(II) organometallic cross-linkers and poly(4-vinylpyridine) in DMSO defines a three-dimensional associative polymer network. Frequency-dependent dynamic mechanical moduli of a series of four different bulk materials, measured across several decades of oscillatory strain rates, are found to be quantitatively related through the pyridine exchange rates measured on model Pd(II) and Pt(II) complexes. Importantly, the mechanism of ligand exchange in the networks is found to be the same solvent-assisted pathway observed in the model complexes, and so the bulk mechanical properties are determined by relaxations that occur when the cross links are dissociated from the polymer backbone. It is how often the cross-links dissociate, independently of how long they remain dissociated, that determines the bulk mechanical properties. The quantitative relationship between bulk materials properties and the kinetics and mechanisms observed in model compounds holds promise for the rational, molecular design of materials with tailored mechanical properties. PMID- 16218644 TI - Low-temperature activation and deactivation of high-Curie-temperature ferromagnetism in a new diluted magnetic semiconductor: Ni2+-Doped SnO2. AB - We report the synthesis of colloidal Ni(2+)-doped SnO(2) (Ni(2+):SnO(2)) nanocrystals and their characterization by electronic absorption, magnetic circular dichroism, X-ray absorption, magnetic susceptibility, scanning electron microscopy, and X-ray diffraction measurements. The Ni(2+) dopants are found to occupy pseudooctahedral Sn(4+) cation sites of rutile SnO(2) without local charge compensation. The paramagnetic nanocrystals exhibit robust high-Curie-temperature (T(C)) ferromagnetism (M(s)(300 K) = 0.8 mu(B)/Ni(2+), T(C) >> 300 K) when spin coated into films, attributed to the formation of interfacial fusion defects. Facile reversibility of the paramagnetic-ferromagnetic phase transition is also observed. This magnetic phase transition is studied as a function of temperature, time, and atmospheric composition, from which the barrier to ferromagnetic activation (E(a)) is estimated to be 1200 cm(-1). This energy is associated with ligand mobility on the surfaces of the Ni(2+):SnO(2) nanocrystals. The phase transition is reversed under air but not under N(2), from which the microscopic identity of the activating defect is proposed to be interfacial oxygen vacancies. PMID- 16218646 TI - Divergent oxidative rearrangements in solution and in a zeolite: distal vs proximal bond cleavage of methylenecyclopropanes. AB - Irradiation of 9,10-dicyanoanthracene (DCA) or p-chloranil in the presence of E-1 benzylidene-2-phenylcyclopropane (E-5) in CH(2)Cl(2) causes E-5 to undergo methylenecyclopropane rearrangement. An adduct, Z-7, between DCA and 5 firmly supports the involvement of a bifunctional trimethylenemethane radical cation. In contrast, incorporation of E-5 into HZSM-5 produces trans,trans-1,4-diphenyl-1,3 butadiene radical cation sequestered in the HZSM-5 interior, tt-8(.+)@HZSM-5, identified by ESR and diffuse reflectance spectroscopy. In addition, low yields of tt-8, its cis,trans-isomer (ct-8), and 1-phenyl-1,2-dihydronaphthalene (9) were isolated from the supernatant solution. The sharp contrast between the photoinduced electron-transfer reaction with photosensitizers in solution and the spontaneous reaction with redox-active acidic zeolite offers the prospect of further zeolite-induced regiodivergent reactions in a range of additional substrates. PMID- 16218647 TI - Specific ion effects on the water solubility of macromolecules: PNIPAM and the Hofmeister series. AB - Aqueous processes ranging from protein folding and enzyme turnover to colloidal ordering and macromolecular precipitation are sensitive to the nature and concentration of the ions present in solution. Herein, the effect of a series of sodium salts on the lower critical solution temperature (LCST) of poly(N isopropylacrylamide), PNIPAM, was investigated with a temperature gradient microfluidic device under a dark-field microscope. While the ability of a particular anion to lower the LCST generally followed the Hofmeister series, analysis of solvent isotope effects and of the changes in LCST with ion concentration and identity showed multiple mechanisms were at work. In solutions containing sufficient concentrations of strongly hydrated anions, the phase transition of PNIPAM was directly correlated with the hydration entropy of the anion. On the other hand, weakly hydrated anions were salted-out through surface tension effects and displayed improved hydration by direct ion binding. PMID- 16218648 TI - Charge-mediated recognition of N-terminal tryptophan in aqueous solution by a synthetic host. AB - The molecular recognition of peptides and proteins in aqueous solution by designed molecules remains an elusive goal with broad implications for basic biochemical research and for sensors and separations technologies. This paper describes the recognition of N-terminal tryptophan in aqueous solution by the synthetic host cucurbit[8]uril (Q8). Q8 is known to form 1:1:1 heteroternary complexes with methyl viologen (MV) and a second aromatic guest. Here, the complexes of Q8.MV with (i) the four natural aromatic alpha-amino acids, (ii) four singly charged tryptophan derivatives, and (iii) four tryptophan-containing tripeptides were characterized by isothermal titration calorimetry, mass spectrometry, and UV-visible, fluorescence, and (1)H NMR spectroscopy. We find that Q8.MV binds Trp-Gly-Gly with high affinity (K(a) = 1.3 x 10(5) M(-1)), with 6-fold specificity over Gly-Trp-Gly, and with 40-fold specificity over Gly-Gly Trp. Analysis of the nine indole-containing compounds suggests that peptide recognition is mediated by the electrostatic charge(s) proximal to the indole, and that the mode of binding is consistent for these compounds. Complex formation is accompanied by the growth of a visible charge-transfer band and the quenching of indole fluorescence. These optical properties, combined with the stability and selectivity of this system, are promising for applications in sensing and separating specific peptides. PMID- 16218649 TI - Functionalization of single-walled carbon nanotubes with well-defined polystyrene by "click" coupling. AB - Covalent functionalization of alkyne-decorated SWNTs with well-defined, azide terminated polystyrene polymers was accomplished by the Cu(I)-catalyzed [3 + 2] Huisgen cycloaddition. This reaction was found to be extremely efficient in producing organosoluble polymer-nanotube conjugates, even at relatively low reaction temperatures (60 degrees C) and short reaction times (24 h). The reaction was found to be most effective when a CuI catalyst was employed in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene as an additive. IR spectroscopy was utilized to follow the introduction and consumption of alkyne groups on the SWNTs, and Raman spectroscopy evidenced the conversion of a high proportion of sp(2) carbons to sp(3) hybridization during alkyne introduction. Thermogravimetric analysis indicated that the polymer-functionalized SWNTs consisted of 45% polymer, amounting to approximately one polymer chain for every 200-700 carbons of the nanotubes, depending on polymer molecular weight. Transmission electron microscopy and atomic force microscopy were utilized to image polymer-functionalized SWNTs, showing relatively uniform polymer coatings present on the surface of individual, debundled nanotubes. PMID- 16218650 TI - The impact of saponins or saponin-containing plant materials on ruminant production--a review. AB - Saponins are steroid or triterpene glycoside compounds found in a variety of plants. Some saponin-containing plants, mainly legumes, have been used as animal feed, but others are toxic. Several studies on the effect of saponins on ruminant production have also been reported. Some in vitro and in vivo experiments that demonstrate the beneficial effects of saponin such as defaunation of the rumen and manipulation of the end products of fermentation are described. Defaunation is the selective removal of protozoa from the rumen microbial ecosystem by a cell membrane cholesterol-saponin interaction, which causes cell rupture. Because protozoa in the rumen cause protein turnover by predating on bacteria, defaunation increases the nitrogen utilization of the ruminant and may lead to an increase in growth, milk, or wool production. The growth-promoting effect was evident in the high roughage diet suggesting that the application of saponins or saponin-containing plant materials may be beneficial for the subsistence farmers in developing countries. Saponins are deglycosylated by rumen microbes. Some sapogenins have been detected in the digestive tract of ruminants; however, the direct action of these compounds on the host animal is still unclear. No information on the effects of saponin on ruminant reproduction is available. There is an urgent need for a systematic evaluation of the most active structural components of the saponins, and their interaction with the microbial community, the host animal, and the diet. Along with these studies, the direct effects of saponins or their microbial degradation products on the host must be examined in order to get the full understanding of the metabolism and beneficial effects of saponins on animals. PMID- 16218651 TI - ACE inhibitory peptides derived from enzymatic hydrolysates of animal muscle protein: a review. AB - Naturally occurring ACE (angiotensin converting enzyme) inhibitory peptides have a potential as antihypertensive components in functional foods or nutraceuticals. These peptides have been discovered in various food sources from plant and animal protein origin. In this paper an overview is presented of the ACE inhibitory peptides obtained by enzymatic hydrolysis of muscle protein of meat, fish, and invertebrates. Some of these peptides do not only show in vitro ACE inhibitory activity but also in vivo antihypertensive activity in spontaneously hypertensive rats. To focus on new sources of ACE inhibitory peptides, more specifically insects and other invertebrates, we compared the vertebrate and invertebrate musculature and analyzed phylogenetic relationships. PMID- 16218652 TI - Separation, characterization, and quantitation of phenolic acids in a little known blueberry (Vaccinium arctostaphylos L.) fruit by HPLC-MS. AB - The aim of this study was the qualitative and quantitative determination of free, ester, glycoside, and ester-bound phenolic acids in the blueberry (Vaccinium arctostaphylos L.) fruit. A method for the determination of the profile of phenolic acids of four different phenolic fractions in the fruit was developed using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Thirteen compounds (gallic, protocatechuic, p-hydroxybenzoic, m-hydroxybenzoic, gentisic, chlorogenic, p-coumaric, caffeic, ferulic, syringic, sinapic, salicylic, and trans-cinnamic acids) were identified and quantified in the berry. These experimental results showed that the predominant phenolic acid in the fruit of V. arctostaphylos is caffeic acid in free and insoluble ester-bound forms and p-coumaric acid in soluble ester and glycoside forms. Seven phenolic acids were identified as hydroxybenzoic acid derivatives (HBAs) and four as hydroxycinnamic acid derivatives (HCAs). Total content of HBAs and HCAs in the four phenolic fractions constituted 30.1 and 69.9% of the free, 27.9 and 72.1% of the ester, 24.7 and 75.3% of the glycoside, and 51.7 and 48.3% of the ester-bound forms, respectively. Total phenolics as the sum of individual phenolic acids identified is 698.5 ng/g of fresh weight (fw) for the free, 3399.2 ng/g of fw for the ester, 3522.1 ng/g of fw for the glycoside, and 3671.6 ng/g of fw for the ester-bound phenolic fractions. The present results were compared with reported levels of phenolic acids in the fruits of different Vaccinium species. These data suggest that the fruit can be considered as a potentially good dietary source of phenolic acids. PMID- 16218653 TI - Kinetics of the surface hydrolysis of raw starch by glucoamylase. AB - The hydrolysis of raw starch catalyzed by glucoamylase has been studied with starch granules of different sizes by use of an amperometric glucose sensor by which the direct and continuous observation of the concentration of glucose can be achieved even in a thick raw starch suspension. The initial rate of the enzymatic hydrolysis in the raw starch suspension increased with increasing concentration of the enzyme to approach a saturation value and was proportional to the amount of substrate. Also, the rate was proportional to the specific surface area of the substrate. The experimental results can be explained well by the rate equations derived from a three-step mechanism, which consists of adsorption of the free enzyme onto the surface of the substrate, reaction of the adsorbed enzyme with the substrate, and liberation of the product. PMID- 16218654 TI - Rapid detection of kernel rots and mycotoxins in maize by near-infrared reflectance spectroscopy. AB - Near-infrared (NIR) spectroscopy is a practical spectroscopic procedure for the detection of organic compounds in matter. It is particularly useful because of its nondestructiveness, accuracy, rapid response, and easy operation. This work assesses the applicability of NIR for the rapid identification of micotoxigenic fungi and their toxic metabolites produced in naturally and artificially contaminated products. Two hundred and eighty maize samples were collected both from naturally contaminated maize crops grown in 16 areas in north-central Italy and from ears artificially inoculated with Fusarium verticillioides. All samples were analyzed for fungi infection, ergosterol, and fumonisin B1 content. The results obtained indicated that NIR could accurately predict the incidence of kernels infected by fungi, and by F. verticillioides in particular, as well as the quantity of ergosterol and fumonisin B1 in the meal. The statistics of the calibration and of the cross-validation for mold infection and for ergosterol and fumonisin B1 contents were significant. The best predictive ability for the percentage of global fungal infection and F. verticillioides was obtained using a calibration model utilizing maize kernels (r2 = 0.75 and SECV = 7.43) and maize meals (r2 = 0.79 and SECV = 10.95), respectively. This predictive performance was confirmed by the scatter plot of measured F. verticillioides infection versus NIR predicted values in maize kernel samples (r2 = 0.80). The NIR methodology can be applied for monitoring mold contamination in postharvest maize, in particular F. verticilliodes and fumonisin presence, to distinguish contaminated lots from clean ones, and to avoid cross-contamination with other material during storage and may become a powerful tool for monitoring the safety of the food supply. PMID- 16218655 TI - Quantitation of clovamide-type phenylpropenoic acid amides in cells and plasma using high-performance liquid chromatography with a coulometric electrochemical detector. AB - A high-performance liquid chromatography (HPLC) method was developed for measuring the concentrations of clovamide-type phenylpropenoic acid amides (N caffeoyldopamine and N-caffeoyltyramine) in cell and plasma samples. The separation was performed on a Nova-Pak C18 column using an isocratic buffer with a coulometric electrochemical detector with four electrode channels. Using the HPLC method, N-caffeoyldopamine and N-caffeoyltyramine could be detected with good peak resolutions at respective retention times (4 and 6.4 min). The calibration curves were linear over the ranges (0.1 and 100 microM), and their lower limit of detection was as little as 100 fmol. For quantifying N caffeoyldopamine and N-caffeoyltyramine in cell and plasma samples, the samples were extracted by extraction methods with more than 95% recoveries. After extraction, the amides were detected with the same sensitivity, peak resolutions, and retention times. Using this method, plasma concentrations of N caffeoyltyramine were determined in blood samples collected at 12, 24, 30, 36, 48, 60, and 75 min after the oral administrations of N-caffeoyltyramine (0.5 mg and 2 mg/30 g body weight). This HPLC method with an electrochemical detector is the first reported method able to quantify N-caffeoyldopamine and N caffeoyltyramine in biological samples with excellent detection limits, peak resolutions, discrete retention times, and consistent reproducibility. PMID- 16218656 TI - Determination of phenolic compounds in Perilla frutescens L. by capillary electrophoresis with electrochemical detection. AB - A method based on capillary electrophoresis with electrochemical detection has been developed to analyze flavonoids and phenolic acids in Perilla frutescens L. for the first time. Catechin, ferulic acid, apigenin, luteolin, rosmarinic acid, and caffeic acid are major important active ingredients in the plant. Operated in a wall-jet configuration, a 300 microm diameter carbon-disk electrode was used as the working electrode, which exhibits a good response at 0.90 V (versus saturated calomel electrode) for the analytes. Under the optimum conditions, the analytes were baseline separated within 20 min in a 100 mmol/L borax buffer (pH 8.7). Notably, excellent linearity was obtained over 3 orders of magnitude with detection limits (S/N = 3) ranging from 2 x 10(-7) to 1 x 10(-6) g/mL for all analytes. This proposed method has been successfully applied to monitor the flavonoids and phenolic acids contents in the leaves and seeds of P. frutescens L. at different growth stages with relatively simple extraction procedures, and the assay results were satisfactory. PMID- 16218657 TI - Dying-arm disease in grapevines: diagnosis of infection with Eutypa lata by metabolite analysis. AB - Dying-arm disease in grapevines, produced by infection with the ascomycete Eutypa lata, is responsible for major production losses in vineyards. Dieback of the shoots and cordon is believed to be due to acetylenic phenol metabolites produced by the fungus. To identify specific metabolites that could potentially be used for diagnosis of infection, eight E. lata isolates were grown in vitro on hot water extracts from grape varieties with various degrees of tolerance to the foliar symptoms of E. lata dieback. HPLC analysis showed that eutypinol was consistently produced in large amounts, together with smaller amounts of methyleutypinol and eulatachromene; eutypine, the putative toxin, was produced solely on Sauvignon Blanc extract and then in only barely detectable amounts. When E. lata isolates from Cabernet Sauvignon and Merlot were grown on identical media, the amounts of metabolites produced differed significantly between isolates but the pattern of metabolites was quite similar, with eutypinol again predominating. The consistent production of eutypinol indicated that this was the most suitable metabolite for which to analyze in order to diagnose the presence of E. lata. Extraction and analysis of grapevine tissues exhibiting symptoms of dieback failed to show the presence of any metabolites. However, when infected cordon sections were placed in water and cultured for 5 days, eutypinol was readily detected in the aqueous solution; metabolites were not produced from uninfected tissue. This provides a method for detection of infected tissue and indicates that the toxic metabolites react at the point of production, disrupting the vascular structure and inhibiting transport of nutrients, rather than being translocated to tissues that exhibit symptoms. PMID- 16218658 TI - Human gene expression as a tool to determine horticultural maturity in a bioactive plant (Echinacea purpurea L. Moench). AB - A phenological study was conducted to determine the impact of harvest maturity on the immune-modulating properties of Echinacea purpurea. The aerial parts of this plant were collected during seven stages of development and were assayed for a common botanical marker for this species, cichoric acid. Plants of selected development stages were also assayed for total polysaccharides and compared for their immune-modulating effects on the THP-1 monocyte/macrophage cell line by means of a gene expression study. Although the concentration of cichoric acid did not change significantly during the course of the study, stage 1 (advanced vegetative) had the highest concentration of total polysaccharides and exhibited the most potent induction activity on immune-modulating cytokines such as interferon-gamma and tumor necrosis factor-alpha. These findings suggest that the use of gene expression may be an effective tool not only to standardize botanical extracts but also to optimize harvest time. PMID- 16218659 TI - Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L. (Cav.) andOriganum vulgare L. AB - Antilisterial activities of Thymbra capitata and Origanum vulgare essential oils were tested against 41 strains of Listeria monocytogenes. The oil of T. capitata was mainly constituted by one component, carvacrol (79%), whereas for O. vulgare three components constituted 70% of the oil, namely, thymol (33%), gamma terpinene (26%), and p-cymene (11%). T. capitata essential oil had a significantly higher antilisterial activity in comparison to O. vulgare oil and chloramphenicol. No significant differences in L. monocytogenes susceptibilities to the essential oils tested were registered. The minimum inhibitory concentration values of T. capitata essential oil and of carvacrol were quite similar, ranging between 0.05 and 0.2 microL/mL. Antioxidant activity was also tested, the essential oil of T. capitata showing significantly higher antioxidant activity than that of O. vulgare. Use of T. capitata and O. vulgare essential oils can constitute a powerful tool in the control of L. monocytogenes in food and other industries. PMID- 16218660 TI - Red chicories as potent scavengers of highly reactive radicals: a study on their phenolic composition and peroxyl radical trapping capacity and efficiency. AB - Eight varieties of Cichorium genus vegetables (five heavily red colored, one red spotted, and two fully green) were investigated for their phenolic content (by HPLC and UV-vis spectrophotometry) and for their antioxidant activity. In particular, the capacity (that is, the amount of trapped peroxyl radicals) and the efficiency (that is, the amount of antioxidant necessary to halve the steady state concentration of peroxyl radicals) were measured. All of the studied chicories are characterized by the presence of a large amount of hydroxybenzoic and hydroxycinnamic acids, whereas the red color is due to cyanidin glycosides. The presence of these phenolics in red chicories confers to them an exceptionally high peroxyl radical scavenging activity in terms of both capacity and efficiency, particularly in their early stage of growth, and makes this popular and low-cost foods comparable or superior to many foods having well-known antioxidant properties such as red wine, blueberry, and tomato. PMID- 16218661 TI - Influence of feeding malt, bread crust, and a pronylated protein on the activity of chemopreventive enzymes and antioxidative defense parameters in vivo. AB - The aim of the present study was to investigate whether feeding of malt, bread crust, and a pronylated albumin modulates chemoprevention enzymes, such as glutathione-S-transferase (GST) and UDP-glucuronyl-transferase (UDP-GT), and antioxidative defense parameters in vivo and whether the intake of these foods rich in Maillard reaction compounds results in an accumulation of compounds formed in in vivo glycation reactions. After quantitation of pronylated lysine in malt and bread crust, male Wistar rats were fed a standard chow supplemented with 28% of protein containing different amounts of casein, bread crust, caraffa malt, or pronyl bovine serum albumin (BSA) for 15 days. GST activity in the kidneys was increased by 18% (p > 0.05) in animals of the bread crust group, while UDP-GT activity was elevated by 27% in the liver of animals administered pronyl-BSA. Contents of tocopherol in plasma were increased by 33, 14, and 14% in the bread crust, malt, and pronyl-BSA group compared to the control group, while the levels of thiobarbituric acid reactive substances were decreased and the total antioxidant capacity was increased. Parameters of endogenous glycation indicated a 32 and 46% higher load of advanced glycation end products in the kidneys after administration of the malt and the pronyl-BSA containing diet. However, the main systemic effects of dietary malt, bread crust, and pronyl-BSA were, for the first time, demonstrated to be the enhanced antioxidant capacity and the particulate increase in chemopreventive enzymes. PMID- 16218662 TI - Antioxidative compounds from the outer scales of onion. AB - Antioxidative compounds were isolated from the methanol extract of dry outer scales of onion (Allium cepa L.). Nine phenolic compounds (1-9) were finally obtained by reversed-phase high-performance liquid chromatography, and their structures were elucidated by NMR and mass spectrometry analyses. They were the six known compounds, protocatechuic acid (1), 2-(3,4-dihydroxybenzoyl)-2,4,6 trihydroxy-3(2H)-benzofuranone (2), quercetin 4'-O-beta-D-glucopyranoside (3), quercetin (5), 4'-O-beta-d-glucopyranoside of quercetin dimer (7), and quercetin dimer (8), and three novel compounds, condensation products of quercetin with protocatechuic acid (4), adduct of quercetin with quercetin 4'-O-beta-D glucopyranoside (6), and quercetin trimer (9). These phenolic compounds were tested for their antioxidant properties using autoxidation of methyl linoleate in bulk phase or free radical initiated peroxidation of soybean phosphatidylcholine in liposomes. The flavonoid compounds having o-dihydroxy substituent in the B ring were shown to be effective antioxidants against nonenzymic lipid peroxidation. PMID- 16218663 TI - Trichothecene production by Trichoderma brevicompactum. AB - Trichoderma brevicompactum, T. viride, T. harzianum, T. atroviride, T. longibrachiatum, T. erinaceum, T. citrinoviride, and Hypocrea lutea were screened for production of trichothecenes after growth on one or several solid and liquid media. Trichothecenes were detected by liquid chromatography combined with online UV/vis spectroscopy and electrospray high-resolution mass spectrometry. T. brevicompactum produced trichodermin and/or harzianum A on all media investigated, with liquid media yielding the largest amounts. Detection of octa 2Z,4E,6E-trienedioic acid in the harzianum-A-producing strains indicated that harzianum A was synthesized directly by esterification of trichodermol with octa 2Z,4E,6E-trienedioic acid. Both the T. viride strain from which trichodermin was originally isolated and the T. harzianum strain from which harzianum A was originally isolated were shown to belong to T. brevicompactum based on four independent criteria: metabolite profiles, micromorphology, macromorphology on yeast extract sucrose agar and potato dextrose agar, and DNA sequences of the ITS1/ITS2 regions of the nuclear ribosomal DNA. PMID- 16218664 TI - Inhibitory effects of a chemically standardized extract from Scutellaria barbata in human colon cancer cell lines, LoVo. AB - Scutellaria barbata (SB) is a medicinal plant that contains flavonone compounds such as scutellarein, scutellarin, carthamidin, isocarthamidin, and wogonin. A functional proteomic approach was used to study the inhibitory effects of a chemically standardized extract from SB in human colon adrencarcinoma, LoVo. In this work, a stable isotope was not used in the proposed method developed. The whole cell lysates from the control and treated cells were digested with trypsin, and the peptides were separated by two-dimensional (cation-exchange and reversed phase) liquid chromatography and tandem mass spectrometry. The differentially expressed proteins identified using the current approach supported the data obtained from cell-cycle analysis with flow cytometry. With flow-cytometry analysis, a significant increase in the sub G1 phase was observed with a higher dose of extract from SB. Our results suggest that the chemically standardized extract from SB can induce cell death in the human colon cancer cell line. Our current work showed that the proposed platform provided a rapid approach to study the molecular mechanism because of the inhibitory effects of different doses of the botanical extracts on LoVo cell lines. This included a network of proteins involved in metabolism, regulation of the cell cycle, and transcription-factor activity. PMID- 16218665 TI - Chinese cabbage extracts and sulforaphane can protect H2O2-induced inhibition of gap junctional intercellular communication through the inactivation of ERK1/2 and p38 MAP kinases. AB - The cruciferous vegetables such as Chinese cabbages and broccoli are known to have anticancer phytochemicals, and the consumption of cruciferous vegetables has been proposed to protect against various cancers. The anticarcinogenic properties of some Chinese cabbage extracts and sulforaphane glucosinolate (SFN) were assessed by examining their ability to prevent the inhibition of gap junctional intercellular communication (GJIC) induced by hydrogen peroxide (H2O2) in WB-F344 normal rat liver epithelial cells. The cells were preincubated with Chinese cabbage extracts and SFN for 24 h followed by cotreatment with cells and H2O2 (750 microM) for 1 h. Chinese cabbage extracts and SFN prevented the inhibition of GJIC and phosphorylation of gap junction protein connexin43 (Cx43) by H2O2 treatment. Chinese cabbage extracts and SFN were able to prevent the inhibition of GJIC through the blocking of Cx43 phosphorylaton and inactivation of ERK 1/2 and p38 MAP kinase. The results suggest that cruciferous vegetables and their components, SFN, may exert the anticancer effect by targeting the GJIC as a functional dietary chemopreventive agent. PMID- 16218666 TI - selective production and characterization of levan by Bacillus subtilis (Natto) Takahashi. AB - To meet the industrial need of an efficient microbial method for increased levan production, Bacillus subtilis (natto) Takahashi, a commercial natto starter for preparing fermented soybeans (natto), was used to produce levan. After cultivation for 21 h, 40-50 mg of levan mL(-1) was produced in medium containing 20% (w/w) sucrose, which was approximately 50% yield on available fructose. The product consisted of two fractions with different molecular masses (1794 and 11 kDa), which were easily separated by fractionation using an ethanol gradient. The products were well characterized by GPC, 13C NMR, and 1H NMR. The various sugars and concentrations, initial pH, fermentation temperature, and agitation speed affected the levan production by B. subtilis (natto) Takahashi. Takahashi strain is the most efficient levan-producing strain among all of the B. subtilis strains tested and, as previously reported, it produced the highest yield of levan in the least time (21 h) under the common cultivation condition. PMID- 16218667 TI - Development and characterization of films based on chemically cross-linked gliadins. AB - The aim of the present work has been to study the possibility of obtaining modified gliadin films with improved water resistance and mechanical properties by means of promoting intermolecular covalent bonds between polypeptide chains. Prior to casting films, formaldehyde, glutaraldehyde, and glyoxal were used to cross-link proteins at concentrations ranging from 1% to 4% (grams per 100 g of protein). Mechanical properties (tensile strength and elongation at break), water vapor permeability, moisture sorption isotherms, and optical properties of the films produced were evaluated as a function of the cross-linker used. Experimental results showed that some properties of gliadin films were considerably modified. Cross-linking improved the water resistance of films, avoiding their disintegration. Their water barrier properties were also enhanced, but their moisture sorption properties remained unchanged. Formaldehyde imparted greater mechanical strength to films than glutaraldehyde or glyoxal, increasing tensile strength values 10-fold. Addition of the cross-linkers at concentrations in excess of 2.5% did not further improve the mechanical or barrier properties. However, modification with glutaraldehyde or glyoxal imparted an increasingly yellowish tint to the films. PMID- 16218668 TI - Conidia of black aspergilli as new biological adsorbents for ochratoxin A in grape juices and musts. AB - Biological removal of ochratoxin A (OTA) by living and heat-treated dead conidia of black Aspergillus isolates representing the species Aspergillus niger, Aspergillus carbonarius, and Aspergillus japonicus in synthetic and natural grape juices was found to be a two-stage phenomenon. Several lines of evidence suggest that the first observed stage was passive, metabolism was not required, and OTA adsorption on conidia of black aspergilli could be involved. This removal was fast, without delay just after conidial inoculation both in synthetic and natural grape juices. Moreover, even nonviable, heat-treated conidia were capable of removing OTA. Finally, no OTA degradation products were detected. In the second observed stage, removal of OTA was linked to degradation by live conidia only. Ochratoxin alpha, a degradation product of OTA, was detected in the medium after incubation for 30 and 14 h for biseriate (A. niger and A. carbonarius) and uniseriate (A. japonicus) black aspergilli, respectively, when well-developed mycelium appeared. Comparisons between the three black Aspergillus isolates tested showed that A. carbonarius detoxified grape juice most effectively. However, this species often produces OTA. A. niger and A. japonicus isolates were also effective and because those species are not systematically OTA producers, they could be interesting for further OTA detoxification processes in grape juices and musts. PMID- 16218669 TI - Effect of the stone content on the quality of plum and cherry spirits produced from mash fermentations with commercial and laboratory yeast strains. AB - To evaluate the influence of stone content on spirit quality from stone fruit, cherry and plum mashes were prepared and fermented with a commercial and a diploid laboratory yeast strain. Fermentation parameters such as sugar content and ethanol production were followed. Despite an initial lag phase in cherry spirits, both yeast strains performed similarly, as substantiated by the determination of specific flavor compounds, ethyl carbamate, and methanol in the mashes and after distillation. The spirits produced were subjected to sensory analyses by trained panels of at least 25 judges. Although mashes retaining the stones could be clearly distinguished from those where the stones had been removed, no significant preference could be attributed to either spirit, indicating that qualities added by the presence of stones during fermentation are largely a matter of personal taste. Interestingly, the yeast strain used for fermentation seemed to have little influence on the spirit quality. PMID- 16218670 TI - Molecular cloning and characterization of a novel 1-aminocyclopropane-1 carboxylate oxidase gene involved in ripening of banana fruits. AB - One novel banana fruit ripening related 1-aminocyclopropane-1-carboxylate (ACC) oxidase gene quite different from ACC oxidase genes from other species was cloned. In contrast to other studies, the polypeptide encoded by this gene, named Mh-ACO1, lacks the putative leucine zipper motif which is conserved in all known ACC oxidases including the other previously reported banana ACC oxidase, Mh-ACO2. The locus consists of two nearly identical paralogous ACC oxidase genes arranged in opposite orientation and separated by a 3.1-kb intergenic region. The has only two introns, at positions identical to , which comprises a coding region interrupted by three introns. The predicted amino acid sequence of Mh-ACO1 shares less than 50% identity to those of ACC oxidase from other climacteric fruits, while that of Mh-ACO2 shows more than 65% homology. When expressed in Saccharomyces cerevisiae -encoded protein possessed the enzyme activity for ethylene conversion. The levels of mRNA corresponding to both and increased during fruit ripening and were induced by exogenous ethylene. We conclude that both and contribute to increased ethylene production in fruits and these two genes are differentially expressed in fruits and other organs in banana. PMID- 16218672 TI - Optimizing dough proofing conditions to enhance isoflavone aglycones in soy bread. AB - Native beta-glucosidase activity in soy bread can convert isoflavone glucosides to aglycones during proofing, and this study determined the time-temperature dependence of this process. Samples were taken every hour for 4 h during proofing at 22, 32, and 48 degrees C to determine beta-glucosidase activity and isoflavone profiles of the dough. After 1-2 h, the beta-glucosidase activity increased 43 84% achieving a plateau value at 22 degrees C but declining when proofed beyond 2 h at 32 degrees C and 48 degrees C. Large increases in aglycones and corresponding decreases in the simple glucosides were observed during proofing. The level of malonyl-glucosides decreased 3-15%, and acetyl-glucosides were fairly constant. The two higher temperatures drove more rapid conversion: 70-73% of simple glucosides in 2-4 h. The extent of conversion in the early proofing periods corresponded to beta-glucosidase activity. The optimum time-temperature protocol was 2 h at 48 degrees C resulting in a rapid, high conversion. PMID- 16218671 TI - Hot water dipping of olives (Olea europaea) for virgin oil debittering. AB - Olives (Olea europaea L.) of the Manzanilla, Picual, and Verdial varieties harvested at the green mature stage of ripening were dipped in hot water at a range of temperatures between 60 and 72 degrees C for 3 min. Immediately after treatment, oils were physically extracted from the olives. Olive heating promotes a reduction of oil bitterness in direct relationship to the temperature used. Fruit heating at > or =60 degrees C for 3 min did not cause significant changes in acidity, UV absorption, peroxide index, and panel test score of the oils obtained but decreased its oxidative stability. Oils extracted from heated fruit showed higher concentrations of chlorophylls and carotenes and lower total phenol content. PMID- 16218673 TI - Influence of lipid extraction process on the rheological characteristics, swelling power, and granule size of rice starches in excess water. AB - The influence of the lipid extraction process on both macroscopic and microscopic characteristics of nonwaxy rice starch gelatinization in excess water was examined. Surface lipids extraction did not change the thermodynamics of starch gelatinization but lead to a significant reduction (33%) in the enthalpy of starch-lipid complex melting at high temperature, resulting in less viscous dispersions. Internal lipid extraction using hot aqueous alcoholic solutions resulted in an irreversible increase in starch granule diameter (50% increase in D[4,3]) and a dramatic change in cooking characteristics of the starch. Instead of the bimodal swelling observed for native nonwaxy rice starch, only one broad transition in swelling, solubility, granule size, and viscosity was observed in the case of the totally defatted starch. While the total removal of lipids resulted in a slight increase in starch swelling at intermediate temperatures, the harshness of the process caused irreparable changes leading to notably lower swelling at high temperatures. PMID- 16218674 TI - Effects of added sodium caseinate on the formation of particles in heated milk. AB - The effects of heat at temperatures in the range of 80-90 degrees C on mixtures of reconstituted skim milk powder (RSMP) and sodium caseinate have been determined. In the absence of caseinate, the action of heat on RSMP produces soluble complexes of whey proteins and kappa-casein, as well as complexes of whey protein with the casein micelles. When sodium caseinate was added to RSMP at levels of 0.5 and 1.0%, the denaturation of the whey protein and the production of the soluble complexes in the serum were hardly affected, either in rate or in amount. However, during the heating, the caseinate disappeared from the serum. Further studies on model mixtures of the different components showed that it was probable that the bulk of the caseinate associated with the casein micelles during heating, probably by binding inside the surface layer of kappa-casein, because no increase in the diameters of the casein micelles could be observed. PMID- 16218675 TI - Red clover Trifolium pratense L. phytoestrogens: UV-B radiation increases isoflavone yield, and postharvest drying methods change the glucoside conjugate profiles. AB - Isoflavone extracts of red clover Trifolium pratense L. (cv. Pawera) with dissimilar glucoside conjugate profiles were obtained by employing different postharvest drying methods. The most prominent isoflavones found were formononetin and biochanin A and their corresponding glucosides and malonyl glucoside esters. Postharvest freeze drying inhibited the conversion of the glycosides to the aglycones, while vacuum drying allowed for maximum conversion of the glycosides to their corresponding aglycones. Air drying produced a low level of the aglycones formononetin and biochanin A, and oven drying promoted decarboxylation of the malonyl glucosides to the acetyl glucosides. Exposure to enhanced UV-B radiation resulted in an increase in total formononetin and biochanin A isoflavone levels, indicating that harvest during a period of high ambient UV-B radiation may be appropriate for maximum yield. The levels of caffeic acid and flavonols also increased by about 40 and 250%, respectively, on exposure to enhanced UV-B radiation. PMID- 16218676 TI - Structure and chemical composition of bast fibers isolated from developing hemp stem. AB - Microscopic and chemical changes of hemp bast fibers were studied during the maturation from vegetative to grain maturity stages at both apical and basal regions of the stems. The content of protein was the main factor related to fiber maturation, whereas increased proportions of mannose and glucose and decreasing levels of galactose were also highly significant. Enhanced glucose deposition in apical fibers could be related to the gradual thickening of the fibers, whereas in basal regions the thickness of the fibers nearly reached the maximum at vegetative stages. In contrast, the extent of lignification remained close to 3 4% during plant growth. Hemp fiber lignins were rich in guaiacyl units and would be rather condensed in nature. In addition, the proportion of p-hydroxyphenyl units displayed a constant decline during maturation. A progressive chemical fractionation of hemp fibers provided further insights to the occurrence and nature of noncellulosic polysaccharides. Notably, these data pointed out that maturation is accompanied by a significant increase in water- and alkali-soluble components containing glucose- and mannose-related polymers and a decrease in arabinose and galactose components disrupted by diluted hydrochloric acid. Taken together, chemical features of the noncellulosic components suggest that the architecture of hemp fibers differs slightly from that of the more widely studied flax fibers. PMID- 16218677 TI - Contents of phenolic acids, alkyl- and alkenylresorcinols, and avenanthramides in commercial grain products. AB - The contents of free and total phenolic acids and alk(en)ylresorcinols were analyzed in commercial products of eight grains: oat (Avena sativa), wheat (Triticum spp.), rye (Secale cerale), barley (Hordeum vulgare), buckwheat (Fagopyrum esculentum), millet (Panicum miliaceum), rice (Oryza sativa), and corn (Zea mays). Avenanthramides were determined in three oat products. Free phenolic acids, alk(en)ylresorcinols, and avenanthramides were extracted with methanolic acetic acid, 100% methanol, and 80% methanol, respectively, and quantified by HPLC. The contents of total phenolic acids were quantified by HPLC analysis after alkaline and acid hydrolyses. The highest contents of total phenolic acids were in brans of wheat (4527 mg/kg) and rye (4190 mg/kg) and in whole-grain flours of these grains (1342 and 1366 mg/kg, respectively). In other products, the contents varied from 111 mg/kg (white wheat bread) to 765 mg/kg (whole-grain rye bread). Common phenolic acids found in the grain products were ferulic acid (most abundant), ferulic acid dehydrodimers, sinapic acid, and p-coumaric acid. The grain products were found to contain either none or only low amounts of free phenolic acids. The content of avenanthramides in oat flakes (26-27 mg/kg) was about double that found in oat bran (13 mg/kg). The highest contents of alk(en)ylresorcinols were observed in brans of rye (4108 mg/kg) and wheat (3225 mg/kg). In addition, whole-grain rye products (rye bread, rye flour, and whole wheat flour) contained considerable levels of alk(en)ylresorcinols (524, 927, and 759 mg/kg, respectively). PMID- 16218678 TI - Novel inhibitors of the mitochondrial respiratory chain: oximes and pyrrolines isolated from Penicillium brevicompactum and synthetic analogues. AB - The capacity of inhibition of the mammalian mitochondrial respiratory chain of brevioxime 5a, a natural insecticide compound isolated from Penicillium brevicompactum culture broth, and another 15 analogue compounds, other oximes 5b and 5c; two diastereomeric pyrrolidines 1c' and 1c' '; five pyrrolines 3c', 3c' ' (diastereomers between them), 3a, 3b, and 6; two oxazines 4c' and 4c' ' (also diastereomers between them); and four pyrrol derivatives 7-10, are analyzed in this paper. Compounds 3b, 3c', 3c' ', 4c', 4c' ', 5b, 5c, 6, and 10 were found to be inhibitors of the integrated electron transfer chain (NADH oxidase activity) in beef heart submitochondrial particles (SMP), establishing that all of them except compound 3b and 6 only affected to complex I of the mitochondrial respiratory chain. The most potent product was 5b, with an IC50 of 0.27 microM, similar to the IC50 values of other known complex I inhibitors. The diastereomeric pairs 1c'/1c' ', 3c'/3c' ', 4c'/4c' ', and 5c have not been previously described. Chemical characterization, on the basis of spectral data, is also shown. PMID- 16218679 TI - Effect of sunscreen and antioxidant on the stability of pyrethrin formulations. AB - The stability of pyrethrins in formulations containing different combinations of antioxidant and sunscreen was studied with model system experiments. The mechanism of disappearance of pyrethrins was mainly due to photodegradation, but some loss by codistillation can occur. In formulations, the presence of mineral oil and emulsifiers determined a decrease on codistillation losses. The presence of the antioxidant in formulations even at high concentrations did not affect pyrethrin photodegradation rates, but rising amounts of sunscreen determined a progressive increase on half-life times of pyrethrins. A combination of sunscreen and antioxidant at low concentrations provided an effective protection against sunlight similar to that obtained by high amounts of sunscreen alone. PMID- 16218680 TI - Chemical pretreatment of olive oil mill wastewater using a metal-organic framework catalyst. AB - Olive oil mill wastewaters (OOMW) are not suited for direct biological treatment because of their nonbiodegradable and phytotoxic compound (such as polyphenols) content. Advanced technologies for treatment of OOMW consider mainly the use of solid catalysts in processes that can be operated at room conditions. A system based on combined actions of catalytic oxidations and microbial technologies was studied. The wet hydrogen peroxide catalytic oxidation (WHPCO) process is one of the new emerging oxidation processes particularly attractive for the pretreatment of highly polluted OOMW containing polyphenols that are not suited for classical treatments. In this work, the biodegradability of OOMW was evaluated before and after treating the wastewater samples by the WHPCO process using a metal-organic framework (MOF) as a catalyst. This material, containing Cu and prepared with benzene-1,3,5-tricarboxylic acid (BTC), is a robust metal-organic polymer with a microporous structure that is reminiscent of the topology of zeolite frameworks. PMID- 16218681 TI - Analysis of volatile compounds and triglycerides of seed oils extracted from different poppy varieties (Papaver somniferum L.). AB - Poppy seed oil (Oleum Papaveris Seminis) is used for culinary and pharmaceutical purposes, as well as for making soaps, paints, and varnishes. Astonishingly, hardly anything was yet known about the volatile compounds of this promising comestible. Likewise, there are no current published data about the triglyceride (TAG) composition of poppy seed oils available. In this investigation solid-phase microextraction (SPME) with DVB/Carboxen/PDMS Stable-Flex fiber was applied to the study of volatile compounds of several seed oil samples from Papaver somniferum L. (Papaveraceae). 1-Pentanol (3.3-4.9%), 1-hexanal (10.9-30.9%), 1 hexanol (5.3-33.7%), 2-pentylfuran (7.2-10.0%), and caproic acid (2.9-11.5%) could be identified as the main volatile compounds in all examined poppy seed oil samples. Furthermore, the TAG composition of these oils was analyzed by MALDI ReTOF- and ESI-IT-MS/MS. The predominant TAG components were found to be composed of linoleic, oleic, and palmitic acid, comprising approximately 70% of the oils. TAG patterns of the different poppy varieties were found to be very homogeneous, showing also no significant differences in terms of the applied pressing method of the plant seeds. PMID- 16218682 TI - Influence of the addition of rosemary essential oil on the volatiles pattern of porcine frankfurters. AB - The effect of the addition of increasing levels of rosemary essential oil (150, 300, and 600 mg/kg) on the generation of volatile compounds in frankfurters from Iberian and white pigs was analyzed using solid-phase microextraction coupled to gas chromatography and mass spectrometry (SPME-GC-MS). Lipid-derived volatiles such as aldehydes (hexanal, octanal, nonanal) and alcohols (pentan-1-ol, hexan-1 ol, oct-1-en-3-ol) were the most abundant compounds in the headspace (HS) of porcine frankfurters. Frankfurters from different pig breeds presented different volatile profiles due to their different oxidation susceptibilities as a likely result of their fatty acid composition and vitamin E content. Rosemary essential oil showed a different effect on the generation of volatiles depending on the type of frankfurter in which they were added. In frankfurters from Iberian pigs, the antioxidant effect of the essential oil improved with increasing levels, showing the highest activity at 600 mg/kg. In contrast, 150 mg/kg of the essential oil improved the oxidative stability of frankfurters from white pigs, whereas higher levels led to no effect or a prooxidant effect. The activity of the essential oil could have been affected by the different fatty acid compositions and vitamin E contents between types of frankfurters. SPME successfully allowed the isolation and analysis of volatile terpenes from frankfurters with added rosemary essential oil including alpha-pinene, beta myrcene, l-limonene, (E)-caryophyllene, linalool, camphor, and 1,8-cineole, which might contribute to the aroma characteristics of frankfurters. PMID- 16218683 TI - Identification of metallic-smelling 1-octen-3-one and 1-nonen-3-one from solutions of ferrous sulfate. AB - Taste threshold tests of ferrous sulfate (FeSO4) solutions have been confounded by the presence of putative odorants. To detect the presence of odorants released from these solutions solid-phase microextraction (SPME) was used to collect volatiles in the headspace above FeSO4 solutions. Gas chromatography-olfactometry of samples collected over three time periods (1, 5, and 16 h) and at two temperatures (22 and 37 degrees C) revealed the presence of several metallic smelling odorants in the headspace. Using authentic standards, two of the odorants were conclusively identified as 1-octen-3-one and 1-nonen-3-one. Trace levels of other odorants were also detected, but dilution experiments indicated that 1-nonen-3-one was at least 10 times more potent than anything else released from the solutions. 1-Octen-3-one and 1-nonen-3-one are excellent candidates for the metallic odor responses often observed in threshold testing of solutions of FeSO4. PMID- 16218684 TI - Dynamic headspace analysis of the release of volatile organic compounds from ethanolic systems by direct APCI-MS. AB - Static equilibrium headspace was diluted with a stream of nitrogen to study the stability of the volatile headspace concentration. The headspace dilution profile of 18 volatile compounds above aqueous and ethanolic solutions was measured in real time using atmospheric pressure chemical ionization-mass spectrometry. Under dynamic conditions the volatiles headspace concentration above water solutions decreased readily upon dilution. The presence of ethanol helped to maintain the volatile headspace concentration when the ethanol solution concentration was above 50 mL/L. This effect was such that under dynamic conditions the absolute volatile concentration above an ethanolic solution was higher than that above an aqueous solution, contrary to results observed in equilibrium studies. The ratio of the headspace concentration of volatiles above ethanolic 120 mL/L and water solutions was correlated to their air/water partition coefficient. PMID- 16218685 TI - Effects of the addition of amino acids and peptides on lipid oxidation in a powdery model system. AB - The effects of the addition of amino acids and peptides on the oxidation of eicosapentaenoic acid ethyl ester (EPE) encapsulated by maltodextrin (MD) were investigated. The encapsulated lipid was prepared in two steps, that is, by mixing of EPE with MD solutions (+/-amino acids and peptides) to produce emulsions and freeze-drying of the resultant emulsions. The addition of amino acids and peptides improved the oxidation stability of EPE encapsulated with MD, and the inhibition of lipid oxidation by the amino acids and peptides was more effective at 70% relative humidity (RH). Met, Arg, and Trp were effective amino acids for antioxidation at RH = 10 and 40%, whereas at RH = 70%, His was the most effective amino acid, preventing the oxidation of EPE almost perfectly. Carnosine also exhibited a strong antioxidant effect at RH = 70%, but the effect of anserine was inferior. The addition of Met + Trp or Met + Arg inhibited the oxidation of EPE encapsulated with MD at RH = 40%. Cys accelerated the oxidation of EPE, indicating that the thiyl radical may act as a pro-oxidant. No close relationship was observed between the radical scavenging abilities of amino acids and peptides measured in the aqueous diphenylpicrylhydrazyl solution and their antioxidative effects in the powdery system. It is possible that the radical scavenging ability of amino acids and peptides detected by ESR in the powder system is responsible for the antioxidative activity of these compounds. PMID- 16218686 TI - Relative contribution of phytates, fibers, and tannins to low iron and zinc in vitro solubility in pearl millet (Pennisetum glaucum) flour and grain fractions. AB - In vitro digestions were performed on pearl millet flours with decreased phytate contents and on two dephytinized or nondephytinized pearl millet grain fractions, a decorticated fraction, and a bran fraction with low and high fiber and tannin contents, respectively. Insoluble residues of these digestions were then incubated with buffer or enzymatic solutions (xylanases and/or phytases), and the quantities of indigestible iron and zinc released by these different treatments were determined. In decorticated pearl millet grain, iron was chelated by phytates and by insoluble fibers, whereas zinc was almost exclusively chelated by phytates. In the bran of pearl millet grain, a high proportion of iron was chelated by iron-binding phenolic compounds, while the rest of iron as well as the majority of zinc were chelated in complexes between phytates and fibers. The low effect of phytase action on iron and zinc solubility of bran of pearl millet grain shows that, in the case of high fiber and tannin contents, the chelating effect of these compounds was higher than that of phytates. PMID- 16218687 TI - Classification of wheat varieties based on structural features of arabinoxylans as revealed by endoxylanase treatment of flour and grain. AB - Arabinoxylans (AX) are cell wall polysaccharides of complex structure involved in many aspects of wheat flour end uses. The study of the variations of AX structure can lead to the identification of genes involved in their biosynthesis, and thus in the control of the various aspects of grain quality related to their presence. A method is proposed to identify AX variations directly in whole grain by enzymatic degradation. An endoxylanase from Trichoderma viride was used to extract AX from a collection of 20 wheat cultivars (Triticum aestivum L.). Enzymatic degradation products were analyzed by HPAEC and multivariate analysis techniques (principal component analysis, canonical correlation analysis, and cluster analysis) were applied to analyze chromatographic data. The method evidenced variations in the proportion of mono- and disubstitution of the xylan backbone by arabinose side chains, allowing classification of the different varieties according to the structural features of AX. A similar classification was obtained starting from flour or whole grain, indicating that the method was specific of AX from endosperm tissues. In conclusion, the method combining endoxylanase treatment of wheat grain and the analysis of degradation products, e.g., enzymatic fingerprinting, can be applied to collections of wheat cultivars, and possibly other cereals in order to establish quantitative trait loci related to the biosynthesis of AX. PMID- 16218688 TI - Carotenoids in three species of Corbicula Clams, Corbicula japonica, Corbicula sandai, and Corbicula sp. (Chinese Freshwater Corbicula Clam). AB - Carotenoids were investigated in three species of corbicula clams, Corbicula japonica, Corbicula sandai, and Corbicula sp. (Chinese freshwater corbicula clam). Forty-three carotenoids were isolated. Among them, 7,8-didehydro-beta cryptoxanthin (12), peridininol 5,8-furanoxide (38), pyrrhoxanthin 5,8-furanoxide (40), and pyrrhoxanthinol 5,8-furanoxide (43) are newly reported as naturally occurring carotenoids. Their structures were characterized on the basis of UV vis, FAB-MS including MS/MS experiments, and 1H NMR spectroscopic data. The total carotenoid contents in C. japonica, C. sandai, and Chinese freshwater corbicula clam were found to be 5.3, 2.6, and 0.3 mg/100 g in the edible part (wet weight), respectively. Peridinin (34) and its derivatives were found to be major carotenoids in C. japonica, which inhabits brackish water. On the other hand, lutein (13) was found to be the major carotenoid in C. sandai and Chinese corbicula clams, which inhabit freshwater. These patterns well reflected the carotenoids in their dietary algae. 7',8'-Didehydrodeepoxyneoxanthin (19), corbiculaxanthin (21), corbiculaxanthin 3'-acetate (22), and 6-epiheteroxanthin (24) were found in all three species of corbicula clams and have not previously been found in other shellfishes. They were assumed to be peculiar carotenoids in corbicula clams. PMID- 16218689 TI - Stability of spray-dried tuna oil emulsions encapsulated with two-layered interfacial membranes. AB - omega-3 Fatty acids have numerous health benefits, but their addition to foods is limited by oxidative rancidity. Spray-drying tuna oil-in-water emulsion droplets with a coating of lecithin and chitosan multilayer system could produce emulsion droplet interfacial membranes that are cationic and thick, both factors that can help control lipid oxidation. Physicochemical and oxidative stability of the spray-dried emulsions were determined as a function of storage temperature and relative humidity (RH). The combination of ethylenediaminetetraacetic acid (EDTA) and mixed tocopherols was able to increase the oxidative stability of dried emulsions. Lipid oxidation was more rapid during storage at low relative humidity (11% and 33% compared to 52% RH). At high moisture, physical modifications in the sample were observed, including reduced dispersibility and formation of brown pigments. Sugar crystallization or Maillard products produced at the higher humidities may have inhibited oxidation. Overall, spray-dried tuna oil-in-water emulsions stabilized by lecithin-chitosan membranes were more oxidatively stable than bulk oils and thus have excellent potential as an omega-3 fatty acid ingredient for functional foods. PMID- 16218690 TI - Effect of copigments and grape cultivar on the color of red wines fermented after the addition of copigments. AB - The prefermentation addition of copigments led to significantly different red wines according to the copigment structure (flavonol or hydroxycinnamic acid) and the grape cultivar [Tempranillo (= Cencibel) or Cabernet Sauvignon]. The flavonol rutin enhanced copigmentation and anthocyanin extraction, improving the red color, but the hydroxycinnamic acids (especially caffeic acid) had converse results. The above effects were higher in Cabernet Sauvignon wines, particularly if rutin or p-coumaric acid was used. These wines showed the highest copigmentation as they contained more anthocyanins and flavonols, whereas the coumaroylated anthocyanins of Tempranillo wines could have prevented the action of the added copigments. After 21 months, the main pyranoanthocyanins found were the malvidin-3-glucoside 4-vinylphenol and the malvidin-3-glucoside 4 vinylcatechol (pinotin A) adducts. The results suggested that the former adduct was primarily generated following enzymatic decarboxylation of p-coumaric acid during fermentation, whereas pinotin A was formed through a pure chemical reaction, which depended on the concentration of free caffeic acid during aging. PMID- 16218691 TI - Relationship of superficial scald development and alpha-farnesene oxidation to reactions of diphenylamine and diphenylamine derivatives in Cv. Granny Smith apple peel. AB - Cv. Granny Smith apple fruit, treated at harvest with aqueous emulsions containing diphenylamine (DPA) and DPA derivatives, were evaluated for the peel disorder superficial scald (scald) after 6 months of cold storage at 1 degrees C plus 0 or 7 days at 20 degrees C. Metabolism of these derivatives and alpha farnesene oxidation were also evaluated after 6 months. Derivatives substituted at the para position prevented scald, but scald developed on fruit treated with derivatives substituted in the amino, ortho, or meta positions. The extent of scald control was also dependent on the chemical nature of the functional group used to derivatize DPA. Hydroxylation of DPA and DPA derivatives during storage was not associated with scald control. Methoxylated DPA derivatives produced during storage resulted from O-methylation of C-hydroxylated derivatives rather than C-methoxylation of DPA. N-Nitrosodiphenylamine provided partial scald control, possibly resulting from its degradation to DPA, indicating that the amino hydrogen of DPA may be crucial for scald control. Results suggest that functional group position and chemical properties both contribute to the efficacy of DPA derivatives for scald control. PMID- 16218692 TI - Influence of the spatial organization of the perimysium on beef tenderness. AB - The spatial distribution of the intramuscular connective tissue (IMCT) in four types of beef muscle (Biceps femoris, Infraspinatus, Longissimus thoracis, and Pectoralis profundus) was examined using histology and magnetic resonance imaging (MRI). The surface and the length of the IMCT and the surface of the myofiber bundles were evaluated by image analysis. The texture of the cooked meat from these muscles was measured both instrumentally by a compression test and by sensory analysis. The relationship between muscle structure and meat texture was studied by general discriminant analysis. The models obtained could assign correctly up to 87% of the samples to two tenderness classes. Histology and MRI provided complementary information about the microscopic and macroscopic IMCT structures, respectively. Both were necessary to predict sensory tenderness whereas only the MRI measurements were necessary to predict instrumental toughness. Tough muscles had smaller MRI myofiber bundles (0.7-1 mm radius) than tender muscles. PMID- 16218693 TI - Effects of lipase, lipoxygenase, peroxidase, and rutin on quality deteriorations in buckwheat flour. AB - To investigate the effects of changes in lipase, lipoxygenase, peroxidase (POX), and rutin concentrations on the quality of buckwheat flour, 14 buckwheat varieties were stored for 0, 4, 10, and 30 days at 5 or 20 degrees C. During the storage period, lipase activity correlated to pH (significantly negative) and water-soluble acid (WSA) (significantly positive). The lipoxygenase 1 protein concentration had a negative correlation to WSA (significant at 0 and 4 storage days at 5 degrees C and at 0 and 10 storage days at 20 degrees C). POX had significant correlation to pH and peroxide value (POV) at 5 degrees C, whereas it was not significant at 20 degrees C. The rutin concentration had negative correlations to WSA (significant at 30 days of storage at 5 degrees C and at 4 days of storage at 20 degrees C). Thus, lipase activity plays an important role that relates to lipid degradation in quality deterioration of buckwheat flour. PMID- 16218694 TI - Levels and trends of polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (PCBs) in Spanish commercial fish and shellfish products, 1995-2003. AB - The polychlorinated dibenzo-p-dioxin (PCDD), dibenzofuran (PCDF), and polychlorinated biphenyl (PCB) contents of 123 Spanish commercial salmon, tuna fish, sardine, oyster, mussel, and clam samples from 1995 to 2003 were investigated. A significant decrease of dioxin and non-ortho PCB concentrations in the studied species was found over the years. The decrease was greater in the case of dioxins than in that of non-ortho PCBs, especially during the early years of the study. PCB and PCDD/F concentrations in the years 2001-2003 were comparable to those reported in the literature for similar species collected after 1999. Mean PCB concentrations ranged from 3.46 ng/g of fresh weight (fw) in clams to 100 ng/g of fw in tuna fish. PCDD/F mean current levels ranged from 0.62 pg/g of fw in clams to 2.89 pg/g of fw in oysters. Toxic equivalent quantities (WHO-TEQ) ranged from 0.05 pg of WHO-TEQ(PCDD/Fs)/g of fw in clams to 0.5 pg of WHO-TEQ(PCDD/Fs)/g of fw in salmon (in the upper bound determination levels). When coplanar PCBs were included, the WHO-TEQ(PCDD/Fs+cop) (PCBs) values increased by a range of 1.7 times in oysters to 14.1 times in tuna fish. The decrease in dioxin concentrations suggests that efforts to control dioxin emissions and to reduce human exposure through foodstuffs are succeeding. The high contribution of PCBs to total WHO-TEQs in the fish and shellfish species investigated suggests that it is important to determine PCBs in foodstuffs, and especially in fish products, and they should be included in further research and future legislation. PMID- 16218695 TI - Model study on the effect of 15 phenolic olive mill wastewater constituents on seed germination and Vibrio fischeri metabolism. AB - Olive mill wastewaters (OMW) can be a severe problem when disposed of as untreated because of their high organic load, elevated concentration of polyphenols, and moderately low biodegradability. In the present study, the acute toxicity of 15 compounds with low molecular weight (<350 Da), catechol, four benzoic acids, three phenylacetic acids, three phenylethanols, and four cinnamic acids, already isolated from the reverse osmosis in the fractionation of OMW, was assessed on the marine bacterium Vibrio fischeri and on the seeds of two dicotyledonous species Cucumis sativus and Lepidium sativum, and on one monocotyledon Sorghum bicolor. Results of phytotoxicity showed that the most toxic compounds were catechol (EC50s ranging from 0.40 mmol/L for S. bicolor to 1.09 for C. sativus) and hydroxytyrosol, (EC50s ranging from 0.47 mmol/L for S. bicolor to 1.55 for C. sativus) while the toxic potential on bacteria was particularly elevated with EC50 values 1 or 2 orders less than phytotoxicity. These results suggested that the risk of OMW disposal may be more elevated for the water compartment than for the soil. PMID- 16218696 TI - Comment on volatile components from mango (Mangifera indica L.) cultivars. PMID- 16218699 TI - Carbon nanotube inner phase chemistry: the Cl- exchange SN2 reaction. AB - Density functional calculations have been carried out to investigate the nature of the inner phase of a (6,6) carbon nanotube, using the Cl(-) exchange S(N)2 reaction as an indicator. Inside the carbon nanotube the classical barrier height increases by 6.6 kcal/mol due to the nanotube polarizability. This suggests that the inner phase environment can be considered a form of solid solvation, offering the possibility of obtaining altered guest properties and reactivity through dielectric stabilization. PMID- 16218700 TI - Multifunctional probe array for nano patterning and imaging. AB - This letter reports the design, fabrication, and testing of a multifunctional scanning probe array for nanoscale imaging and patterning. The probe array consists of multiple cantilever probes, with each probe being able to perform a dedicated function such as scanning probe lithography (e.g., dip pen nanolithography and scanning probe contact printing) or scanning probe microscopy (e.g., atomic force microscopy and lateral force microscopy). The bending states of each probe can be controlled by using an integrated thermal electric actuator so that it is possible to engage any individual probe(s) independently for writing or imaging purposes. The multifunctional probe array is therefore capable of performing a rich variety of operations with minimal chemical crosstalk and high registration accuracy. It will eliminate the need for probe chip exchanges and increase the operational efficiency. The probe tips in a given array may be made of different materials. Further, the tip and cantilever may be made of different materials for a given probe. In this work, we focus on the development of a probe array consisting of dip pen nanolithography probes, scanning probe contact printing probes (of various tip sizes), and scanning probe microscopy probes. PMID- 16218701 TI - Photoluminescence intermittency of InGaAs/GaAs quantum dots confined in a planar microcavity. AB - Photoluminescence intermittency, or "blinking", was observed in semiconductor InGaAs/GaAs quantum dots (QDs) inside a planar microcavity. Most of the blinking QDs were found around defect sites such as dislocation lines naturally formed in the GaAs barrier layers, and the carrier traps responsible for blinking had an excitation threshold of approximately 1.53 eV. The blinking properties of epitaxial QDs and colloidal nanocrystal QDs were also compared by performing laser intensity dependent measurements and statistics of the "on" and "off" time distributions. PMID- 16218702 TI - Self-regulated structures in nanocomposites by directed nanoparticle assembly. AB - Adding surface-modified silica nanoparticles (NPs) to polymer blend films, we demonstrate that directed interfacial segregation of NPs stabilize either three dimensional (3D) interpenetrating or 2D discrete structures at high and low volume fractions of NPs, respectively. A simple interfacial energy argument provides a general guideline for predicting whether the NPs are directed to the interface between phases or into one phase. The final morphology and domain size can be predicted from the volume fraction of NPs, film thickness, and NP size. PMID- 16218703 TI - Stretching DNA using the electric field in a synthetic nanopore. AB - The mechanical properties of DNA over segments comparable to the size of a protein-binding site (3-10 nm) are examined using an electric-field-induced translocation of single molecules through a nanometer diameter pore. DNA, immersed in an electrolyte, is forced through synthetic pores ranging from 0.5 to 1.5 nm in radius in a 10 nm thick Si(3)N(4) membrane using an electric field. To account for the stretching and bending, we use molecular dynamics to simulate the translocation. We have found a threshold for translocation that depends on both the dimensions of the pore and the applied transmembrane bias. The voltage threshold coincides with the stretching transition that occurs in double-stranded DNA near 60 pN. PMID- 16218704 TI - Novel optical surface properties of Ca2+ -doped MgO nanocrystals. AB - Surface doping of oxide nanoparticles is important in fields ranging from heterogeneous catalysis to optoelectronics. Here, we report the solvent-free synthesis of mixed calcium-magnesium oxide nanocrystals in the size range between 5 and 40 nm. Although CaMgO mixtures are thermodynamically forbidden on a macroscopic scale, Calcium ions can be distributed homogeneously in MgO using chemical vapor deposition (CVD) as a nonequilibrium technique. Subsequent thermal activation leads to calcium ion segregation into the nanocrystal surface which, in addition to the synthesis parameters, provides efficient means for manipulating the optical surface properties of insulating oxide nanocrystals. A novel material with unexpected photonic behavior, such as enhanced photoluminescence emission which is also red-shifted with respect to those of CaO and MgO, was found. PMID- 16218705 TI - Switching the conformation of a DNA molecule with a chemical oscillator. AB - pH oscillations generated by a nonequilibrium chemical reaction are used to switch a pH-sensitive DNA structure between two distinct conformations. The utilization of a chemical oscillator represents a novel method for achieving autonomous motion in molecular devices. The oscillatory reaction is a variant of the Landolt reaction and produces pH variations in the range between pH 5 and 7. In this range, a cytosine-rich DNA strand can be switched between a random coil conformation and the folded i-motif structure. The conformational changes are monitored simultaneously with the pH value in fluorescence-resonance energy transfer experiments. PMID- 16218706 TI - Electrochemical fabrication of nanodimensional multilayer films. AB - A novel method of fabricating nanodimensional multilayer films using electrochemistry is described. A thin layer of tantalum (Ta) is sputtered on a smooth insulating substrate. Ta is partially electrochemically oxidized (anodized) forming a Ta(2)O(5) layer. The rate of Ta consumption, the rate of Ta(2)O(5) expansion, and the dependence of Ta(2)O(5) thickness on anodization conditions have been carefully characterized to enable accurate predictions of the resulting thicknesses of both layers. Due to strong planarization action of the anodization process, the resulting interfaces Ta/Ta(2)O(5) and Ta(2)O(5)/electrolyte are remarkably smooth. The next layer of Ta is deposited on top of Ta(2)O(5), and the process is repeated as many times as needed. The Ta(2)O(5) layers are amorphous and pinhole free. We report fabrication of 10 layer structures with pitches ranging from 200 nm down to 12 nm and with excellent uniformity between the layers. The smallest achieved thickness of Ta layers is only 2.8 +/- 0.1 nm. The edges of such films, after proper polishing and etching, could serve as templates in nanoimprint lithography and in other applications. PMID- 16218707 TI - Detecting single stranded DNA with a solid state nanopore. AB - Voltage biased solid-state nanopores are used to detect and characterize individual single stranded DNA molecules of fixed micrometer length by operating a nanopore detector at pH values greater than approximately 11.6. The distribution of observed molecular event durations and blockade currents shows that a significant fraction of the events obey a rule of constant event charge deficit (ecd) indicating that they correspond to molecules translocating through the nanopore in a distribution of folded and unfolded configurations. A surprisingly large component is unfolded. The result is an important milestone in developing solid-state nanopores for single molecule sequencing applications. PMID- 16218708 TI - Size sorting of protein assemblies using polymeric gradient surfaces. AB - We report on a novel approach for the size-dependent fractionation of protein assemblies on polymeric surfaces. Using a simple temperature gradient method to generate one-dimensional gradients of grafted poly(ethylene glycol), we fabricated silicon-oxide chips with a gradually changing surface density of kinesin motor molecules. We demonstrate that such a bioactive surface can be used to sort gliding microtubules according to their length. To our knowledge, this is the first example of the self-organized sorting of protein assemblies on surfaces. PMID- 16218709 TI - Multilevel, room-temperature nanoimprint lithography for conjugated polymer-based photonics. AB - We demonstrate the multilevel patterning of organic light-emitting polymers by room-temperature nanoimprint lithography (RT-NIL), which is impossible to obtain by conventional hot embossing. In particular, we realize one- and two-dimensional photonic crystals with 500 nm periodic features and investigate the changes in the optical properties (luminescence and quantum yield) of the organic active layer. An increase of the quantum yield by 2.4% for the patterned film with respect to the untextured one and the enhancement of the output light emitted at a particular angle (Theta = 69 degrees) are observed for gratings whose Bragg periodicity matched the emission wavelength of the polymer. The employment of RT NIL to pattern polymer semiconductors without degradation of their optical properties represents a strategic route for the realization of novel nanopatterned optoelectronic devices. PMID- 16218710 TI - Phonon confinement effects in hybrid virus-inorganic nanotubes for nanoelectronic applications. AB - Genetically modified viruses have been proposed recently as templates for the assembly of nanometer-scale components of electronic circuits. Here we show that, in addition to their role as nanotemplates, viruses can actually improve the electron transport properties in semiconductor nanotubes grown on them. In the considered hybrid virus-inorganic nanostructures, which consist of silica or silicon nanotubes deposited on tobacco mosaic viruses, the confined acoustic phonons are found to be redistributed between the nanotube shell and the acoustically soft virus enclosure. As a result, the low-temperature electron mobility in the hybrid virus-silicon nanotube can increase by a factor of 4 compared to that of an empty silicon nanotube. Our estimates also indicate an enhancement of the low-temperature thermal conductivity in the virus-silicon nanotube, which can lead to improvements in heat removal from the hybrid nanostructure-based nanocircuits. PMID- 16218711 TI - Nanoscale rapid melting and crystallization of semiconductor thin films. AB - The current study details nanosecond laser-based rapid melting and crystallization of thin amorphous silicon (a-Si) films at the nanoscale using two different optical near-field processing schemes. Both apertureless and tapered fiber near-field scanning optical microscope probes were utilized to deliver highly confined irradiation on the target surface. The various modification regimes produced as a result of the rapid a-Si melting and crystallization transformations were shown to critically depend on the applied laser fluence. Consequently, the crystallized pattern morphology and feature size could be finely controlled. High energy density was observed to impart ablation surrounded by a narrow melt ring. At much lower incident laser energy density, single nanostructures with a lateral dimension of approximately 90 nm were defined. PMID- 16218713 TI - Expanding the optical trapping range of gold nanoparticles. AB - We demonstrate stable three-dimensional (3D) single-beam optical trapping of gold nanoparticles with diameters between 18 and 254 nm. Three-dimensional power spectral analysis reveals that, for nanoparticles with diameters less than 100 nm, the trap stiffness is proportional to the volume of the particle. For larger particles, the trap stiffness still increases with size, however, less steeply. Finally, we provide numbers for the largest forces exertable on gold nanoparticles. PMID- 16218712 TI - Patterned assembly of genetically modified viral nanotemplates via nucleic acid hybridization. AB - The patterning of nanoparticles represents a significant obstacle in the assembly of nanoscale materials and devices. In this report, cysteine residues were genetically engineered onto the virion surface of tobacco mosaic virus (TMV), providing attachment sites for fluorescent markers. To pattern these viruses, labeled virions were partially disassembled to expose 5' end RNA sequences and hybridized to virus-specific probe DNA linked to electrodeposited chitosan. Electron microscopy and RNAase treatments confirmed the patterned assembly of the virus templates onto the chitosan surface. These findings demonstrate that TMV nanotemplates can be dimensionally assembled via nucleic acid hybridization. PMID- 16218714 TI - Growth and optical properties of strained GaAs-GaxIn 1-x P core-shell nanowires. AB - We have synthesized GaAs-Ga(x)In(1-x)P (0.34 < x < 0.69) core-shell nanowires by metal-organic vapor phase epitaxy. The nanowire core was grown Au-catalyzed at a low temperature (450 degrees C) where only little growth takes place on the side facets. The shell was added by growth at a higher temperature (600 degrees C), where the kinetic hindrance of the side facet growth is overcome. Photoluminescence measurements on individual nanowires at 5 K showed that the emission efficiency increased by 2 to 3 orders of magnitude compared to uncapped samples. Strain effects on the band gap of lattice mismatched core-shell nanowires were studied and confirmed by calculations based on deformation potential theory. PMID- 16218715 TI - Single molecular imaging and spectroscopy of conjugated polyelectrolytes decorated on stretched aligned DNA. AB - DNA is the prototype template for building nanoelectronic devices by self assembly. The electronic functions are made possible by coordinating electronic polymer chains to DNA. This paper demonstrates two methods for fabrication of aligned and ordered DNA nanowires complexed with conjugated polyelectrolytes (CPEs). The complex can be formed either in solution prior to stretching or after stretching of the bare DNA on a surface. Molecular combing was used to stretch the complexes on surface energy patterned surfaces, and PMMA for the bare DNA. Single molecular spectroscopy, in fluorescence, and microscopy, in atomic force microscopy, give evidence for coordination of the short CPE chains to the aligned DNA. PMID- 16218717 TI - Organometallic spintronics: dicobaltocene switch. AB - A single-molecule spintronic switch and spin valve using two cobaltocene moieties is proposed. Spin-dependent transport through a lead-molecule-lead junction has been calculated using first-principles density functional and nonequilibrium Green function methods. We find that the antiparallel (singlet) configuration of the cobaltocene spins blocks electron transport near the Fermi energy, while the spin parallel (triplet) configuration enables much higher current. The energy difierence between the antiparallel and parallel states depends on the insulating spacer separating the two cobaltocenes, allowing switching through the application of a moderate magnetic field. PMID- 16218716 TI - Elastic property of vertically aligned nanowires. AB - An atomic force microscopy (AFM) based technique is demonstrated for measuring the elastic modulus of individual nanowires/nanotubes aligned on a solid substrate without destructing or manipulating the sample. By simultaneously acquiring the topography and lateral force image of the aligned nanowires in the AFM contacting mode, the elastic modulus of the individual nanowires in the image has been derived. The measurement is based on quantifying the lateral force required to induce the maximal deflection of the nanowire where the AFM tip was scanning over the surface in contact mode. For the [0001] ZnO nanowires/nanorods grown on a sapphire surface with an average diameter of 45 nm, the elastic modulus is measured to be 29 +/- 8 GPa. PMID- 16218718 TI - Surface plasmon standing waves in large-area subwavelength hole arrays. AB - A flexible and parallel procedure to generate large-area, free-standing films of subwavelength hole arrays has been demonstrated. This method is materials general, and multilayered films of different materials were constructed. The optical quality of these films was tested using a near-field scanning optical microscope, which revealed the formation of surface plasmon standing wave patterns that were consistent with numerical simulations. Because the properties of the holes and the film materials can be easily tailored, new types of plasmonic and photonic devices can be envisioned and tested. PMID- 16218719 TI - Nanoscale, phonon-coupled calorimetry with sub-attojoule/Kelvin resolution. AB - We have developed an ultrasensitive nanoscale calorimeter that enables heat capacity measurements upon minute, externally affixed (phonon-coupled) samples at low temperatures. For a 5 s measurement at 2 K, we demonstrate an unprecedented resolution of DeltaC approximately 0.5 aJ/K ( approximately 36 000 k(B)). This sensitivity is sufficient to enable heat capacity measurements upon zeptomole scale samples or upon adsorbates with sub-monolayer coverage across the minute cross sections of these devices. We describe the fabrication and operation of these devices and demonstrate their sensitivity by measuring an adsorbed (4)He film with optimum resolution of approximately 3 x 10(-5) monolayers upon an active surface area of only approximately 1.2 x 10(-9) m(2). PMID- 16218720 TI - The structures of Ag55+ and Ag55- : trapped ion electron diffraction and density functional theory. AB - We report the experimental structure determination of cold, mass selected Ag(55)(+/-) cluster ions using the recently developed technique of trapped ion electron diffraction. By comparison of experimental and theoretical molecular scattering functions and consideration of computed total energies, we show that Ag(55)(+) constitutes an ideal Mackay icosahedron, whereas Ag(55)(-) is a weakly Jahn-Teller distorted icosahedron. Isomers of other structural types, for example, decahedral or close-packed, could be ruled out. The candidate structures were obtained by density functional theory calculations. PMID- 16218721 TI - Electric field tuning of plasmonic response of nanodot array in liquid crystal matrix. AB - In this work we demonstrate the feasibility of electric-field tuning of the plasmonic spectrum of a novel gold nanodot array in a liquid crystal matrix. As opposed to previously reported microscopically observed near-field spectral tuning of individual gold nanoparticles, this system exhibits macroscopic far field spectral tuning. The nanodot-liquid crystal matrix also displays strong anisotropic absorption characteristics, which can be effectively described as a collective ensemble within a composite matrix in the lateral dimension and a group of noninteracting individual particles in the normal direction. The effective medium model and the Mie theory are employed to describe the experimental results. PMID- 16218722 TI - Mechanical properties of ZnS nanobelts. AB - Mechanical properties of ZnS nanobelts were measured at room temperature by direct nanoindentation experiments. It was found that the ZnS nanobelts achieve 79% increase in hardness but 52% decrease in elastic modulus compared to bulk ZnS. The nanobelts were found to exhibit creep under indentation. Indentation cracking was preferred along the belt growth direction. Indentation deformation behavior and fracture mechanisms of the ZnS nanobelts are discussed in conjunction with their crystalline structure, size effect, and surface-to-volume ratio. PMID- 16218723 TI - Magnetomicelles: composite nanostructures from magnetic nanoparticles and cross linked amphiphilic block copolymers. AB - We report the synthesis, characterization, and covalent surface chemistry of "magnetomicelles", cross-linked, amphiphilic block-copolymer micelles that encapsulate superparamagnetic iron oxide nanoparticles. Because these composite nanostructures assemble spontaneously from solution by simultaneous desolvation of nanoparticle and amphiphilic poly(styrene(250)-block-acrylic acid(13)) components, explicit surface functionalization of the particles is not required, and the encapsulation method was applied to different magnetic nanoparticle sizes and compositions. TEM images of the magnetomicelles illustrated that the number of encapsulated particles could be dictated rationally by synthetic conditions. The magnetic properties of the particles were characterized by SQUID magnetometry and followed the general Langevin magnetic model for superparamagnetic materials. The micellar shells of these particles were functionalized using covalent chemistry that would not ordinarily be possible on the magnetic particle surface. As a result, this noncovalent approach provides a new route to technological applications of hydrophobic magnetic nanomaterials that lack appropriate conjugate surface chemistry. PMID- 16218724 TI - Nanoscale wear and machining behavior of nanolayer interfaces. AB - An atomic force microscope was used to subnanometer incise a nanomultilayer to consequently expose individual nanolayers and interfaces on which sliding and scanning nanowear/machining have been performed. The letter reports the first observation on the nanoscale where (i) atomic debris forms in a collective manner, most-likely by deformation and rupture of atomic bonds, and (ii) the nanolayer interfaces possess a much higher wear resistance (desired for nanomachines) or lower machinability (not desired for nanomachining) than the layers. PMID- 16218725 TI - Bottom-up growth of carbon nanotube multilayers: unprecedented growth. AB - An unusual growth phenomenon, with no precedent in vapor-phase thin film growth, is described here, for the case of the growth of stacked multiple layers of vertically aligned carbon nanotubes(1-6) on solid substrates. As multiple layers of ordered nanotubes are sequentially deposited from the vapor onto the substrate, each layer nucleates and grows from the original substrate surface at the bottom of the existing multiple stacks of nanotubes. In contrast to conventional understanding of thin film deposition,(7) the mechanism here has similarities to porous oxide film formation on surfaces.(8) The stacked layers of aligned nanotubes act as fully permeable membranes for the downward diffusion of growth precursor vapors, allowing growth to occur at the buried solid interface. The preexisting multiple nanotube stacks lift up to accommodate the vertical growth of fresh layers, allowing the formation of nanotube towers extending in millimeter lengths. Our results provide evidence for a new growth phenomenon, characterized by selective, interface-driven, bottom-up growth of self-assembled nanowires at buried interfaces, covered with weakly adhering thick porous membranes. PMID- 16218726 TI - Tailoring aqueous solubility of functionalized single-wall carbon nanotubes over a wide pH range through substituent chain length. AB - Carboxylic acid-functionalized SWNTs prepared via the reaction of an amino acid, NH2(CH2)nCO2H, with fluoronanotubes show similar levels of sidewall functionalization; however, the solubility in water is controlled by the length of the hydrocarbon side chain (i.e., n). The 6-aminohexanoic acid derivative is soluble in aqueous solution (0.5 mg mL(-1)) between pH 4 and 11, whereas the glysine and 11-aminoundecanoic acid derivatives are insoluble across all pH values. PMID- 16218727 TI - Ultraviolet electroluminescence from ZnO/polymer heterojunction light-emitting diodes. AB - We report ultraviolet electroluminescence at 390 nm from diode structures consisting of electrodeposited ZnO nanorods sandwiched between a transparent SnO(2) film and a p-type conducting polymer. The nanorods are embedded in an insulating polystyrene layer. ZnO deposition occurs at 90 degrees C and produces vertically oriented nanorods with very high uniformity over areas of approximately 20 cm(2). Electron diffraction shows the nanorods to be single crystalline wurtzite ZnO. As-grown films show a broad electroluminescence band over the visible spectrum. Annealing at moderate temperatures (T = 300 degrees C) increases the emission and strongly raises the excitonic contribution. Optimally processed films show a narrow ultraviolet electroluminescence line at approximately 390 nm. PMID- 16218728 TI - Plasmons in the metallic nanoparticle-film system as a tunable impurity problem. AB - We show that the plasmon resonances of a metallic nanoparticle interacting with the surface plasmons of a metallic film is an electromagnetic analogue of the spinless Anderson-Fano model. This is the same model used to describe the interaction of a localized electronic state with a continuous band of electronic states. The three characteristic regimes of this model are realized here, where the energy of the nanoparticle plasmon resonance lies above, within, or below the energy band of surface plasmon states. These three interaction regimes are controlled by film thickness. The latter regime is experimentally observed and identified. PMID- 16218729 TI - Oriented mesoporous organosilicate thin films. AB - Coassemblies of block copolymers and inorganic precursors offer a path to ordered inorganic nanostructures. In thin films, these materials combined with domain alignment provide highly robust nanoscopic templates. We report a simple path to control the morphology, scaling, and orientation of ordered mesopores in organosilicate thin films through the coassembly of a diblock copolymer, poly(styrene-b-ethylene oxide) (PS-b-PEO), and an oligomeric organosilicate precursor that is selectively miscible with PEO. Continuous films containing cylindrical or spherical pores are generated by varying the mixing composition of symmetric PS-b-PEO and an organosilicate precursor. Tuning interfacial energy at both air/film and film/substrate interfaces allows the control of cylindrical pore orientation normal to the supported film surfaces. Our method provides well ordered mesoporous structures within organosilicate thin films that find broad applications as highly stable nanotemplates. PMID- 16218730 TI - Encoding morphology in oxide nanostructures during their growth. AB - Programmable control over the overall structure of SnO(2) nanowires grown by vapor-solid synthesis is shown to be possible by pulse modulating the flow rate of the carrier gas in which oxygen (one of the reactants) is entrained. The control is shown to depend on the local oscillation of the supersaturation condition for the SnO vapor (another reactant) in the vicinity of the growing nanostructure. The latter triggers dramatic, reproducible oscillations in the lateral dimensions of the nanostructure and in the direction of its growth. The method provides a means for producing predictable morphological and compositional variations in 1D nanostructures, thereby potentially resulting in a high yield of custom-designed nanostructures. PMID- 16218731 TI - Surface combustion microengines based on photocatalytic oxidations of hydrocarbons at room temperature. AB - The concept of a surface combustion microengine that is fuelled by volatile hydrocarbons at room temperature is demonstrated on a microcantilever covered with a thin layer of titanium oxide (TiO(2)). Exposing this microengine to ultraviolet (UV) radiation and hydrocarbon vapor produces controlled bending of the microcantilever as a result of differential stress produced by photocatalytic oxidation of organic molecules on the TiO(2) coating. Compared to the motion generated solely by UV radiation or hydrocarbon adsorption, the unique photocatalytic-mechanical effects in the presence of UV and hydrocarbon produce more work and exhibit fast response. The surface combustion based microengines would require less maintenance in minimally controlled field environment and could be potentially used in construction of miniature movable machines, conversion of solar and chemical energy to mechanical work, when extended to a large array of microcantilevers. We believe such microengines can be fuelled by a variety of molecules or mixtures due to the generally favorable photocatalytic reactivity of TiO(2), thus potentially offering a broad approach for mechanical work generation from multiple energy sources. PMID- 16218732 TI - Disorder effects in focused-ion-beam-deposited Pt contacts on GaN nanowires. AB - The current-bias (I-V) characteristics at various temperatures, T, of focused-ion beam (FIB)-deposited Pt contacts on GaN nanowires evolves from low-resistance ohmic (linear I-V) to rectifying as the diameter increases, and both exhibit strongly nonmetallic T-dependence. The small-diameter (66 nm) T-dependent resistance is explained by two-dimensional variable range hopping with a small characteristic energy, ensuring low resistance at 300 K. For large diameters (184 nm), back-to-back Schottky barriers explain the nonlinear I-V at all T values and permit an estimate of doping concentration from the bias-dependent barrier height. Both behaviors can be understood by accounting for the role of FIB induced amorphization of GaN underneath the contact, as confirmed by cross sectional transmission electron microscopy. PMID- 16218733 TI - Localized surface plasmon resonance spectroscopy of single silver nanocubes. AB - In this work, we use dark-field microscopy to observe a new plasmon resonance effect for a single silver nanocube in which the plasmon line shape has two distinct peaks when the particles are located on a glass substrate. The dependence of the resonance on nanocube size and shape is characterized, and it is found that the bluer peak has a higher figure of merit for chemical sensing applications than that for other particle shapes that have been studied previously. Comparison of the measured results with finite difference time domain (FDTD) electrodynamics calculations enables us to confirm the accuracy of our spectral assignments. PMID- 16218734 TI - Shape memory effect in Cu nanowires. AB - A rubber-like pseudoelastic behavior is discovered in single-crystalline face centered-cubic (FCC) Cu nanowires in atomistic simulations. Nonexistent in bulk Cu, this phenomenon is associated primarily with a reversible crystallographic lattice reorientation driven by the high surface-stress-induced internal stresses due to high surface-to-volume ratios at the nanoscale level. The temperature dependence of this behavior leads to a shape memory effect (SME). Under tensile loading and unloading, the nanowires exhibit recoverable strains up to over 50%, well beyond the typical recoverable strains of 5-8% for most bulk shape memory alloys (SMAs). This behavior is well-defined for wires between 1.76 and 3.39 nm in size over the temperature range of 100-900 K. PMID- 16218735 TI - Wave function engineering in elongated semiconductor nanocrystals with heterogeneous carrier confinement. AB - We explore two routes to wave function engineering in elongated colloidal CdSe/CdS quantum dots, providing deep insight into the intrinsic physics of these low-dimensional heterostructures. Varying the aspect ratio of the nanoparticle allows control over the electron-hole overlap (radiative rate), and external electric fields manipulate the interaction between the delocalized electron and the localized hole. In agreement with theory, this leads to an exceptional size dependent quantum confined Stark effect with field induced intensity modulations, opening applications as electrically switchable single photon sources. PMID- 16218736 TI - Multihydroxylated [Gd@C82(OH)22]n nanoparticles: antineoplastic activity of high efficiency and low toxicity. AB - [Gd@C82(OH)22]n particles (22 nm in a saline solution) of a dose level as low as 10(-7) mol/kg exhibit a very high antineoplastic efficiency ( approximately 60%) in mice. A dose increment of 1 x 10(-7) mol/kg increases the tumor inhibition rate 26%. [Gd@C82(OH)22]n particles have a strong capacity to improve immunity and interfere with tumor invasion in normal muscle cells, nearly without toxicity in vivo and in vitro. Unlike conventional antineoplastic chemicals, the high antitumor efficiency of nanoparticles is not due to toxic effects to cells because they do not kill the tumor cells directly and only about 0.05% of the used dose is found in the tumor tissues. Results suggest that fullerene derivatives with proper surface modifications and sizes may help realize the dream of tumor chemotherapeutics of high-efficacy and low-toxicity. PMID- 16218737 TI - Optical properties of Pd-Ag and Pt-Ag nanoboxes synthesized via galvanic replacement reactions. AB - Silver nanocubes dispersed in water were transformed into Pd-Ag or Pt-Ag nanoboxes by adding either Na(2)PdCl(4) or Na(2)PtCl(4). By controlling the amount of noble metal salt added, and therefore the molar ratio of Na(2)PdCl(4) or Na(2)PtCl(4) to Ag, we could tune the surface plasmon resonance peak of the nanostructures across the entire visible spectrum, from 440 to 730 nm. Replacement of Ag with Pd resulted in the formation of a nanobox composed of a Pd Ag alloy single crystal, but the nanobox formed after replacement of Ag with Pt was instead composed of distinct Pt nanoparticles. DDA calculations suggest that both nanoboxes absorb light strongly, with Q(abs)/Q(sca) approximately 5. After galvanic replacement, Pd-Ag and Pt-Ag nanostructures remain SERS active, suggesting their use as a SERS probe for studying the dependence of interfacial chemistry on composition. PMID- 16218738 TI - Bioconjugated superstructures of CdTe nanowires and nanoparticles: multistep cascade Forster resonance energy transfer and energy channeling. AB - Nanoparticle/nanowire assemblies with a degree of radial organization were prepared around luminescent semiconducting CdTe nanowires using bioconjugation with streptavidin and D-biotin linkers. Red-emitting nanowires (6.62 +/- 1.55 nm diameter, 512 +/- 119 nm length) and green-emitting nanoparticles (3.2 +/- 0.7 nm diameter) were surface-modified with biotin, while orange-emitting nanoparticles (4.1 +/- 1.2 nm diameter) were decorated with streptavidin. CdTe nanocrystals produced two fuzzy layers around the nanowires in which the diameter of CdTe nanoparticles decreased with the distance from the nanowire axis. Forster resonance energy transfer (FRET) from the outside layer of nanoparticles to the central nanowire was observed for nanowires conjugated with 4.1 nm CdTe. Addition of 3.2 nm CdTe resulted in a red-orange-green optical progression with band gaps of CdTe decreasing toward the axis of the superstructure. In this case, 4-fold luminescence enhancement of the nanowire luminescence was observed and was attributed to multistep FRET. This observation indicated the accumulation of photogenerated excitons in the cascade terminal. A simple model of multiconjugated superstructure with cascade energy transfer is developed and used to describe and understand the experimental data. The experimental data and theoretical model suggest the possibility of utilization of the prepared superstructures with radial symmetry in several classes of optoelectronic devices including nanomaterials for energy collection. They can also be a convenient model object for the investigation of methods of energy funneling in nanoscale assemblies. PMID- 16218739 TI - Metal-induced assembly of a semiconductor island lattice: Ge truncated pyramids on Au-patterned Si. AB - We report the two-dimensional alignment of semiconductor islands using rudimentary metal patterning to control nucleation and growth. In the Ge on Si system, a square array of submicron Au dots on the Si (001) surface induces the assembly of deposited Ge adatoms into an extensive island lattice. Remarkably, these highly ordered Ge islands form between the patterned Au dots and are characterized by a unique truncated pyramidal shape. A model based on patterned diffusion barriers explains the observed ordering and establishes general criteria for the broader applicability of such a directed assembly process to quantum dot ordering. PMID- 16218740 TI - Catalytically grown carbon nanotubes of small diameter have a high Young's modulus. AB - Experimental studies of carbon nanotubes (CNTs) obtained through different synthesis routes show considerable variability in their mechanical properties. The strongest CNTs obtained so far had a high Young's modulus of 1 TPa but could only be produced in gram scale quantities. The synthesis by catalytic chemical vapor deposition, a method that holds the greatest potential for large-scale production, gives CNTs with a high defect density. This leads to low Young's modulus values below 100 GPa for multiwall CNTs. Here we performed direct measurements of the mechanical properties of catalytically grown CNTs with only a few walls and find a Young's modulus of 1 TPa. This high value is confirmed for CNTs grown under two different growth conditions where the synthesis parameters such as the hydrocarbon source, catalyst material, and the synthesis temperature were varied. The results indicate that the observed difference in the Young's modulus for the catalytically grown CNTs with high and low numbers of walls is probably related to the growth mechanism of CNT. PMID- 16218741 TI - Charging process and Coulomb-force-directed printing of nanoparticles with sub 100-nm lateral resolution. AB - This article reports on a new charging process and Coulomb-force-directed assembly of nanoparticles onto charged surface areas with sub-100-nm resolution. The charging is accomplished using a flexible nanostructured thin silicon electrode. Electrical nanocontacts have been created as small as 50 nm by placing the nanostructured electrode onto an electret surface. The nanocontacts have been used to inject charge into 50 nm sized areas. Nanoparticles were assembled onto the charge patterns, and a lateral resolution of 60 nm has been observed for the first time. A comparison of the nanoparticle patterns with the surface potential distribution recorded by Kelvin probe force microscopy (KFM) revealed a mismatch in the lateral resolution. One possible explanation is that nanoparticles may visualize charge patterns at a sub-60-nm length scale that is not well resolved using KFM. PMID- 16218742 TI - Immobilization of active hydrogenases by encapsulation in polymeric porous gels. AB - Hydrogenases encapsulated in porous polymeric silica gels retain significant levels of hydrogen production activity when compared to hydrogenases in solution using reduced methyl viologen as an electron donor. Encapsulated hydrogenases remain active after storage at room temperature for longer than four weeks and are less sensitive to proteolytic digestion. Nanoscopic confinement of active hydrogenases in solids paves the way for their potential use in hydrogen producing catalytic materials applications. PMID- 16218743 TI - Slow vibrations in transport through molecules. AB - We show how one can measure the signal from slow jumps of a single molecule between metastable positions using a setup where the molecule is fixed to one lead and one of the coupling strengths is controlled externally. Such a measurement yields information about slow processes deforming the molecule in times much longer than the characteristic time scales for the electron transport process. PMID- 16218744 TI - Assembly and interaction of Au/C core-shell nanostructures: in situ observation in the transmission electron microscope. AB - Using transmission electron microscopy, we identify the temperature-dependent interaction pathway of carbon-supported Au nanoparticles. At low temperature (room temp. to 400 degrees C), Au nanoparticles predominantly interact by coalescence initiated by an atomic Au bridge. At high temperature (400-800 degrees C), the particles assemble into Au/C core-shell nanostructures. C-shells around individual nanoparticles passivate their surface and prevent coalescence. Ultimately coalescence does occur via rupture of the passivating shells, which invariably follows the assembly of C sheets enveloping and compressing multiple closely spaced Au/C nanoparticles. PMID- 16218745 TI - Superhydrophobicity and superhydrophilicity of regular nanopatterns. AB - The hydrophilicity, hydrophobicity, and sliding behavior of water droplets on nanoasperities of controlled dimensions were investigated experimentally. We show that the "hemi-wicking" theory for hydrophilic SiO(2) samples successfully predicts the experimental advancing angles and that the same patterns, after silanization, become superhydrophobic in agreement with the Cassie-Baxter and Wenzel theories. Our model topographies have the same dimensional scale of some naturally occurring structures that exhibit similar wetting properties. Our results confirm that a forest of hydrophilic/hydrophobic slender pillars is the most effective superwettable/water-repellent configuration. It is shown that the shape and curvature of the edges of the asperities play an important role in determining the advancing angles. PMID- 16218746 TI - Light-induced switching between structural forms with different optical properties in a single gallium nanoparticulate. AB - In a single gallium nanoparticulate, self-assembled (from an atomic beam) in a nanoaperture at the tip of a tapered optical fiber, we have observed reversible light-induced reflectivity changes associated with a sequence of transformations between a number of structural forms with different optical properties, stimulated by optical excitation at nanowatt power levels. The ability to change the optical properties of a nanoparticulate using structural transformations provides a new mechanism for photonic functionality on the nanoscale. PMID- 16218747 TI - Nanoscale deformation mechanisms in bone. AB - Deformation mechanisms in bone matrix at the nanoscale control its exceptional mechanical properties, but the detailed nature of these processes is as yet unknown. In situ tensile testing with synchrotron X-ray scattering allowed us to study directly and quantitatively the deformation mechanisms at the nanometer level. We find that bone deformation is not homogeneous but distributed between a tensile deformation of the fibrils and a shearing in the interfibrillar matrix between them. PMID- 16218748 TI - Dielectrophoretically controlled fabrication of single-crystal nickel silicide nanowire interconnects. AB - We report here on applying electric fields and dielectric media to achieve controlled alignment of single-crystal nickel silicide nanowires between two electrodes. Depending on the concentration of nanowire suspension and the distribution of electrical field, various configurations of nanowire interconnects, such as single, chained, and branched nanowires were aligned between the electrodes. Several alignment mechanisms, including the induced charge layer on the electrode surface, nanowire dipole-dipole interactions, and an enhanced local electrical field surrounding the aligned nanowires are proposed to explain these novel dielectrophoretic phenomena of one-dimensional nanostructures. This study demonstrates the promising potential of dielectrophoresis for constructing nanoscale interconnects using metallic nanowires as building blocks. PMID- 16218750 TI - Possible involvement of the RNAi pathway in trinucleotide repeat expansion diseases. AB - A new molecular mechanism of trinucleotide expansion diseases is suggested. The mechanism involves the formation of double-helical RNA hairpins by transcripts carrying (CNG)(n) sequences, which are processed via the RNAi pathway with subsequent RNA silencing of genes containing (CNG)(n) sequences. Depletion of proteins encoded by these genes leads to the specific disease phenotype. The available data on human myotonic dystrophy 1, which results from the (CTG)(n) expansion, support the hypothesis. PMID- 16218751 TI - Protein sequence modules. AB - Conserved protein sequence segments are commonly believed to correspond to functional sites in the protein sequence. A novel approach is proposed to profile the changing degree of conservation along the protein sequence, by evaluating the occurrence frequencies of all short oligopeptides of the given sequence in a large proteome database. Thus, a protein sequence conservation profile can be plotted for every protein. The profile indicates where along the sequences the potential functional (conserved) sites are located. The corresponding oligopeptides belonging to the sites are very frequent across many prokaryotic species. Analysis of a representative set of such profiles reveals a common feature of all examined proteins: they consist of sequence modules represented by the peaks of conservation. Typical size of the modules (peak-to-peak distance) is 25-30 amino acid residues. PMID- 16218752 TI - Regiospecificity of human cytochrome P450 1A1-mediated oxidations: the role of steric effects. AB - Cytochrome P450 1A1 oxidizes a diverse range of substrates, including the procarcinogenic xenobiotic benzo[a]pyrene (B[a]P) and endogenous fatty acid precursors of prostaglandins, such as arachidonic acid (AA) and eicosapentaenoic acid (EA). We have investigated the extent to which enzyme-substrate interactions govern regio- and stereoselectivity of oxidation of these compounds by using docking and molecular dynamics (MD) simulations to examine the likelihood of substrate oxidation at various sites. Due to structural differences between the substrates analyzed, B[a]P and its diols (planar, rigid), and the fatty acids AA and EA (long, flexible), different docking strategies were required. B[a]P, B[a]P 7,8-diols, (+) 7S,8S- and (-) 7R,8R-diols, were docked into the active site of a homology model of P450 1A1 using an automated routine, Affinity (Accelrys, San Diego, CA). AA and EA, on the other hand, required a series of restrained MD simulations to obtain a variety of productive binding modes. All complexes were evaluated by MD-based in silico site scoring to predict product profiles based on certain geometric criteria, such as angle and distance of a given substrate atom from the ferryl oxygen. For all substrates studied, the in vitro profiles were generally reflected by the in silico scores, which suggests that steric factors play a key role in determining regiospecificity in P450 1A1-mediated oxidations. We have also shown that molecular dynamics simulations may be very useful in determination of product profiles for structurally diverse substrates of P450 enzymes. PMID- 16218753 TI - Conformational changes induced in the human protein translin and in the single stranded oligodeoxynucleotides d(GT)(12) and d(TTAGGG)(5) upon binding of these oligodeoxynucleotides by translin. AB - Translin is a human single-stranded DNA and RNA binding protein that has been highly conserved in eukaryotic evolution. It consists of eight subunits having a highly helical secondary structure that assemble into a ring. The DNA and the RNA are bound inside the ring. Recently, some of us demonstrated that the human translin specifically binds the single-stranded microsatellite repeats, d(GT)(n), the human telomeric repeats, d(TTAGGG)(n), and the Tetrahymena telomeric repeats, d(GGGGTT)(n). These data suggested that translin might be involved in recombination at d(GT)(n).d(AC)(n) microsatellites and in telomere metabolism. Other data indicated that translin might stimulate binding of telomerase to single-stranded telomeric overhangs by unwinding secondary structures formed by the telomeric repeats. Here we present a circular dichroism (CD) analysis of complexes formed between the human translin and the microsatellite and telomeric oligodeoxynucleotides d(GT)(12) and d(TTAGGG)(5). We report that conformational changes occur in both the translin and the oligodeoxynucleotides upon formation of the complexes. In translin octamers bound to the oligodeoxynucleotide d(GT)(12), the fraction of alpha-helices decreases from approximately 67% to approximately 50%, while the fraction of turns and of the unordered structure increases from approximately 11% to approximately 17% and from approximately 19% to approximately 24%, respectively. In the bound oligodeoxynucleotide d(GT)(12), we observed CD shifts which are consistent with a decrease of base stacking and a putative anti-syn switch of some guanines. The oligodeoxynucleotide d(TTAGGG)(5) formed intramolecular quadruplexes under the conditions of our assays and translin was found to unfold the quadruplexes into structures consisting of a single hairpin and three unwound single-stranded d(TTAGGG) repeats. We suggest that such unfolding could account for the stimulation of telomerase activity by translin mentioned above. PMID- 16218754 TI - Conformational preferences of the HIV-1 principal neutralizing determinant. AB - The model describing the conformational properties of the HIV-1 principal neutralizing determinant in the geometric space of dihedrals was generated in terms of NMR spectroscopy data published in literature. To gain an object in view, the following successive steps were put into effect: (i) the NMR-based local structures for the HIV(MN) V3 loop were determined in water and in a mixed water/trifluoroethanol (TFE) solvent (7:3), (ii) in either case, the conformations of its irregular segments were analyzed and the secondary structure elements identified, (iii) to appreciate the degree of conformational mobility of the stretch of interest, the simulated structures were compared with each other, (iv) to detect the amino acids retaining their conformations inside the diverse HIV-1 isolates, the structures computed were collated with the one derived previously for the V3 loop from Thailand isolate, and (v) as a matter of record, the structurally rigid residues, that may present the forward-looking targets for AIDS drug researches, were revealed. Summing up the principal results arising from these studies, the following conclusions were drawn: I. The HIV(MN) V3 loop offers the highly mobile fragment of gp120 sensitive to its environment whose changes trigger the large-scale structural reforms, bringing in substantial altering the secondary structure of this functionally important site of the virus envelope. II. In water, it exhibits extended site 1-14 separated by double beta turn 15-20 with unordered region 21-35. III. Adding the TFE gives rise to destruction of the regular structure in the V3 loop N-terminal, stimulates the formation of 3(10)-helix in site 24-31, and affects also its central region 20-25 forming the HIV-1 immunogenic crown. IV. Regardless of statistically significant differences between local structures of the HIV(MN) V3 loop in water and in water/TFE solution, over one-third of residues keeps their conformational states; the register of these amino acids comprises Asn-25 critical for virus binding with primary cell receptor CD4 as well as Arg-3 critical for utilization of CCR5 coreceptor. V. There are no conserved structural motifs within the V3 loops from Minnesota and Thailand HIV-1 strains. However, perceptible portion of amino acids (more than 35%), including those appearing in the functionally important regions of gp120, holds the values of dihedral angles in which case. The implications are discussed in conjunction with the data on the experimental observations for the HIV-1 principal neutralizing determinant. PMID- 16218755 TI - Assessment of a rigorous transitive profile based search method to detect remotely similar proteins. AB - Profile-based sequence search procedures are commonly employed to detect remote relationships between proteins. We provide an assessment of a Cascade PSI-BLAST protocol that rigorously employs intermediate sequences in detecting remote relationships between proteins. In this approach we detect using PSI-BLAST, which involves multiple rounds of iteration, an initial set of homologues for a protein in a 'first generation' search by querying a database. We propagate a 'second generation' search in the database, involving multiple runs of PSI-BLAST using each of the homologues identified in the previous generation as queries to recognize homologues not detected earlier. This non-directed search process can be viewed as an iteration of iterations that is continued to detect further homologues until no new hits are detectable. We present an assessment of the coverage of this 'cascaded' intermediate sequence search on diverse folds and find that searches for up to three generations detect most known homologues of a query. Our assessments show that this approach appears to perform better than the traditional use of PSI-BLAST by detecting 15% more relationships within a family and 35% more relationships within a superfamily. We show that such searches can be performed on generalized sequence databases and non-trivial relationships between proteins can be detected effectively. Such a propagation of searches maximizes the chances of detecting distant homologies by effectively scanning protein "fold space". PMID- 16218756 TI - Oxetane locked thymidine in the Dickerson-Drew dodecamer causes local base pairing distortions -- an NMR structure and hydration study. AB - The introduction of a North-type sugar conformation constrained oxetane T block, 1-(1',3'-O-anhydro-beta-D-psicofuranosyl) thymine, at the T(7) position of the self-complementary Dickerson-Drew dodecamer, d[(5' C(1)G(2)C(3)G(4)A(5)A(6)T(7)T(8)C(9)G(10)C(11)G(12)-3')](2), considerably perturbs the conformation of the four central base pairs, reducing the stability of the structure. UV spectroscopy and 1D NMR display a drop in melting temperature of approximately 10 degrees C per modification for the T(7) oxetane modified duplex, where the T(7) block has been introduced in both strands, compared to the native Dickerson-Drew dodecamer. The three dimensional structure has been determined by NMR spectroscopy and has subsequently been compared with the results of 2.4 ns MD simulations of the native and the T(7) oxetane modified duplexes. The modified T(7) residue is found to maintain its constrained sugar- and the related glycosyl torsion conformations in the duplex, resulting in staggered and stretched T(7).A(6) and A(6).T(7) non-linear base pairs. The stacking is less perturbed, but there is an increased roll between the two central residues compared to the native counterpart, which is compensated by tilts of the neighboring base steps. The one dimensional melting profile of base protons of the T(7) and T(8) residues reveals that the introduction of the North type sugar constrained thymine destabilizes the core of the modified duplex, promoting melting to start simultaneously from the center as well as from the ends. Temperature dependent hydration studies by NMR demonstrate that the central T(7).A(6)/A(6).T(7) base pairs of the T(7) oxetane modified Dickerson-Drew dodecamer have at least one order of magnitude higher water exchange rates (correlated to the opening rate of the base pair) than the corresponding base pairs in the native duplex. PMID- 16218757 TI - Hydration and conformational transitions in DNA, RNA, and mixed DNA-RNA triplexes studied by gravimetry and FTIR spectroscopy. AB - We have studied by gravimetric measurements and FTIR spectroscopy the hydration of duplexes and triplexes formed by combinations of dA(n), dT(n), rA(n), and rU(n) strands. Results obtained on hydrated films show important differences in their hydration and in the structural transitions which can be induced by varying the water content of the samples. The number of water molecules per nucleotide (w/n) measured at high relative humidity (98% R.H.) is found to be 21 for dA(n).dT(n) and 15 for rA(n).rU(n). Addition of a third rU(n) strand does not change the number of water molecules per nucleotide: w/n=21 for rU(n)*dA(n).dT(n) and w/n=15 for rU(n)*rA(n).rU(n). On the contrary, the addition of a third dT(n) strand changes the water content but in a different way, depending whether the duplex is DNA or RNA. Thus, a loss of four water molecules per nucleotide is measured for dT(n)*dA(n).dT(n) while an increase of two water molecules per nucleotide is observed for dT(n)*rA(n).rU(n). The final hydration is the same for both triplexes (w/n=17). The desorption profiles obtained by gravimetry and FTIR spectroscopy are similar for the rA(n).rU(n) duplex and the rU(n)*rA(n).rU(n) triplex. On the contrary, the desorption profiles of the dA(n).dT(n) duplex and the triplexes formed with it (rU(n)*dA(n).dT(n) and dT(n)*dA(n).dT(n)) are different from each other. This is correlated with conformational transitions induced by varying the hydration content of the different structures, as shown by FTIR spectroscopy. Modifications of the phosphate group hydration and of the sugar conformation (S to N type repuckering) induced by decrease of the water content are observed in the case of triplexes formed on the dA(n).dT(n) duplex. PMID- 16218758 TI - Electrostatic potentials of E.coli genome DNA. AB - Distribution of electrostatic potential of the complete sequence of E. coli genome was calculated. Comparative analysis of electrostatic patterns for 359 promoter and nonpromoter nucleotide sequences was carried out. It is found that nonpromoter regions are characterized by more homogeneous distribution of electrostatic potential with no common specific elements. Electrostatic patterns of promoter DNAs can be specified due to the presence of some distinctive motifs which may be involved as promoter signal elements in RNA-polymerase-promoter recognition. PMID- 16218759 TI - Thr-Val-Thr dansyl hydrazide: the first fluorescent tripeptide preferentially binding with at pairs in DNA. AB - In 1991-1993 we amended a DNA-protein recognition model advanced in 1975. Here we test our assumptions with a specially designed tripeptide L-Thr-L-Val-L-Thr-NH-NH Dns (Dns is 5-dimethylaminonaphthalene-1-sulfonic acid residue). It is shown to dimerize in solution (as evidenced by the nonlinear concentration dependence of its fluorescence) and to bind with DNA mainly in beta-dimeric form (S-shaped adsorption isotherm obtained by equilibrium dialysis). The tripeptide is bound in the DNA minor groove (whence it can be displaced with distamycin A), and such complexes become able to associate into 'biduplex' structures (nonlinear dependence of the linear dichroism of bound peptide on DNA concentration). The peptide dimers clearly prefer the AT pairs [half-saturating peptide concentrations are (0.6-0.7) x 10(-4) M for poly(dA).poly(dT) but exceed (2.5 2.8) x10(-4) M for poly(dG).poly(dC)]. These results agree nicely with our earlier suggestions. Since Dns-tagged trivaline has been shown to prefer the GC pairs, we think it now becomes possible to design oligopeptides that would specifically bind to any predefined nucleotide sequence. PMID- 16218760 TI - The cat's "MEOW": new mnemonic for diagnosis and management of falls in the elderly. PMID- 16218761 TI - Medicare Part D prescription coverage: what it means for primary care physicians. PMID- 16218762 TI - Solitary, enlarging red lesion on upper back. Patient appears fine otherwise, except for Crohn's disease. PMID- 16218763 TI - Late-life mania: assessment and treatment of late-life manic symptoms. PMID- 16218764 TI - Assessing falls in older adults: a comprehensive fall evaluation to reduce fall risk in older adults. AB - Falls account for significant morbidity and mortality in the older adult population. A comprehensive fall evaluation (CFE) is proposed, particularly for use in recurrent fallers, those who seek medical attention for a fall, and those with a gait and/or balance disorder. The CFE focuses on key factors in the medical and fall history, review of systems, and physical examination. Interventions utilize a multifactorial model, although balance exercise may be the most critical component. While some components may be marginally successful when presented individually as an intervention (such as correction of vision impairment or environmental hazard reduction), when presented together, fall risk can be significantly reduced. PMID- 16218765 TI - Diagnosing epilepsy in older adults: what does it mean for the primary care physician? AB - Acute and chronic seizures (epilepsy) occur frequently in older adults. However, despite their high prevalence, the condition is often overlooked or misdiagnosed because seizures mimic other common spells in the aged. The causes of seizures in older adults are often the sequelae of highly prevalent conditions that occur in this age group, eg, stroke, dementia, and head trauma. Recognizing and correctly diagnosing seizures can lead to a number of effective treatments. This article reviews the causes and presentation of seizures in older adults and offers an approach for diagnosis. PMID- 16218768 TI - Sources of contraceptive commodities for users in Nigeria. AB - BACKGROUND: Understanding the sources of contraceptive commodities is an important aspect of the delivery of family planning services and is required by planning programme managers for strategic planning purposes. Findings from the 2003 Nigeria Demographic and Health Survey have previously showed that the private sector was the most frequently reported source of contraceptive supply, providing contraception to two and a half times as many women as the public sector. We conducted a community-based study to examine further the sources of contraceptive commodity for users in Nigeria with a view to identifying their preferences for distribution centres. This information would be useful to improve commodity distribution and to build the necessary capacity for satisfactory delivery of contraceptives. METHODS AND FINDINGS: A multi-stage random sampling technique was used. A state was randomly selected to represent each of the four health zones in Nigeria. Two local government areas (LGAs) were then selected representing both urban and rural areas. Ten enumeration areas were subsequently selected from each LGA. Of the 2,001 respondents aged 15-49 years, 1,647 (82.3%) were sexually active, out of which 244 were found to be using contraceptive methods at the time of the study, giving a contraceptive prevalence of 14.8%. The commonest source of information on contraceptives was through friends (34%), followed by the radio (11.5%) and husbands (10.2%). Most respondents procured their contraceptives from chemist/patent medicine shops (19.7%), while only 0.8% obtained them from designated family planning clinics. The younger groups in this study (15-24 years), single people, Catholics, and Muslims, showed a greater preference for chemist/patent medicine shops for their sources of contraceptives. The older groups and married respondents, however, made use of government and private hospitals to obtain their contraceptives. CONCLUSION: Strategies to increase contraceptive use must take into consideration these identified sources of contraceptives with a view to enhancing the quality, quantity, and variety of methods available, and to building capacity for effective service delivery. There is also a need to encourage the establishment of adolescent-friendly clinics where young people can go for counselling and obtain contraceptives of their choice, including emergency contraceptive pills. PMID- 16218766 TI - Improvement of tactile discrimination performance and enlargement of cortical somatosensory maps after 5 Hz rTMS. AB - Repetitive transcranial magnetic stimulation (rTMS) is increasingly used to investigate mechanisms of brain functions and plasticity, but also as a promising new therapeutic tool. The effects of rTMS depend on the intensity and frequency of stimulation and consist of changes of cortical excitability, which often persists several minutes after termination of rTMS. While these findings imply that cortical processing can be altered by applying current pulses from outside the brain, little is known about how rTMS persistently affects learning and perception. Here we demonstrate in humans, through a combination of psychophysical assessment of two-point discrimination thresholds and functional magnetic resonance imaging (fMRI), that brief periods of 5 Hz rTMS evoke lasting perceptual and cortical changes. rTMS was applied over the cortical representation of the right index finger of primary somatosensory cortex, resulting in a lowering of discrimination thresholds of the right index finger. fMRI revealed an enlargement of the right index finger representation in primary somatosensory cortex that was linearly correlated with the individual rTMS induced perceptual improvement indicative of a close link between cortical and perceptual changes. The results demonstrate that repetitive, unattended stimulation from outside the brain, combined with a lack of behavioral information, are effective in driving persistent improvement of the perception of touch. The underlying properties and processes that allow cortical networks, after being modified through TMS pulses, to reach new organized stable states that mediate better performance remain to be clarified. PMID- 16218769 TI - A measles epidemic threshold in a highly vaccinated population. AB - BACKGROUND: Mass vaccination against measles has successfully lowered the incidence of the disease and has changed the epidemic pattern from a roughly biennial cycle to an irregular sequence of outbreaks. A possible explanation for this sequence of outbreaks is that the vaccinated population is protected by solid herd immunity. If so, we would expect to see the fraction of susceptible individuals remaining below an epidemic threshold. An alternative explanation is the occurrence of occasional localised lapses in herd immunity that allow for major outbreaks in areas with a low vaccine coverage. In that case, we would expect the fraction of susceptible individuals to exceed an epidemic threshold before outbreaks occur. These two explanations for the irregular sequence of measles outbreaks can be tested against observations of both the fraction of susceptible individuals and infection attack rates. METHODS AND FINDINGS: We have estimated both the fraction of susceptible individuals at the start of each epidemic year and the infection attack rates for each epidemic year in the Netherlands over a 28-y period. During this period the vaccine coverage averaged 93%, and there was no sustained measles transmission. Several measles outbreaks occurred in communities with low vaccine coverage, and these ended without intervention. We show that there is a clear threshold value for the fraction of susceptible individuals, below which only minor outbreaks occurred, and above which both minor and major outbreaks occurred. A precise, quantitative relationship exists between the fraction of susceptible individuals in excess of this threshold and the infection attack rate during the major outbreaks. CONCLUSION: In populations with a high but heterogeneous vaccine coverage, measles transmission can be interrupted without establishing solid herd immunity. When infection is reintroduced, a major outbreak can occur in the communities with low vaccine coverage. During such a major outbreak, each additional susceptible individual in excess of the threshold is associated with almost two additional infections. This quantitative relationship offers potential for anticipating both the likelihood and size of future major outbreaks when measles transmission has been interrupted. PMID- 16218770 TI - Human health risks from low-level environmental exposures: no apparent safety thresholds. PMID- 16218772 TI - The quality of procedural rural medical practice in Australia. AB - INTRODUCTION: While rural Australians receive much of their procedural medical care from local health professionals in local hospitals, the current workforce shortages, rural economic decline and poor rural health care status all pose challenges to the quality of care they receive. Rural doctors struggle to receive appropriate procedural skills training, rural hospitals struggle to maintain experienced procedurally skilled nurses and other health professionals, and medical equipment, and patients are increasingly referred by clinical protocols to larger urban hospitals. On the other hand, many rural communities value highly their local rural hospital, and advocate the maintenance of hospital services close to home, even though they will have to travel for more specialised services. This article reports an exploration of the quality of a range of clinical cases gathered from rural procedural medical practice. METHODS: The Australian College of Rural and Remote Medicine (ACRRM) approached all trained and procedurally practising rural doctors among their membership. A total of 49 agreed to participate, but only 24 were successful in the prospective recruitment of patients and contribution of patient material (operating theatre notes, anaesthetic records etc) from cases involving general surgery, anaesthetics and obstetrics, the three commonest procedural disciplines in rural medical practice. One of the researchers interviewed patients before and after their procedures and, where available, a family member and a nurse at the hospital. Thus a series of 91 detailed patient case studies was available for analysis These case studies were reviewed from up to four different perspectives: (i) rural doctor peers; (ii) regionalist specialists in the respective discipline; (iii) a medical administrator; and (iv) a rural consumer representative. A thematic analysis of transcribed interviews was conducted. RESULTS: The collected cases represented a range of procedures commonly provided in rural hospitals, although there were relatively few surgical procedures and there was a bias in all three specialty areas towards relatively simple procedures. No adverse outcomes were reported, although some comments, particularly from the rural doctor peers, were made about the need for further information and, in a small number of cases, possible variance from accepted practice. The views of the reviewers substantially agreed that the cases were of average to high quality, although the specialist reviewers were less likely to rate care as 'excellent' than other reviewers. While the comments of the medical reviewers were more technical in nature, the comments of patients and their families, and of the rural consumer reviewer, focussed more on issues such as accessibility, cost and interpersonal communication. Many patients and some nursing staff expressed concern about the sustainability of friendly and accessible local services in the face of workforce shortages and pressure to downgrade rural hospitals. CONCLUSION: This study shows that, where staff and facilities in rural hospitals are accredited for procedural care, there is little evidence of any difference in the quality of that care provided when compared with care expected in urban hospitals. PMID- 16218774 TI - Gene therapy of storage disorders by retroviral and lentiviral vectors. PMID- 16218775 TI - I. Poloxamer-formulated plasmid DNA-based human cytomegalovirus vaccine: evaluation of plasmid DNA biodistribution/persistence and integration. AB - Preclinical studies were conducted in mice and rabbits to evaluate biodistribution/persistence and potential integration of plasmid DNA (pDNA) after intramuscular administration of a poloxamer-formulated pDNAbased vaccine, VCL CT01, encoding gB, pp65, and IE1 human cytomegalovirus (hCMV) immunogens. Tissue distribution in mice vaccinated with VCL-CT01 was compared with that in mice vaccinated with a phosphate- buffered saline (PBS)-formulated control pDNA vaccine. Residual pDNA copy number (PCN), in selected tissues collected on days 3, 30, and 60 after vaccination, was measured by quantitative polymerase chain reaction. In VCL-CT01-vaccinated mice and in control pDNA-vaccinated mice, pDNA was below the limit of detection by day 60 in all tissues except the injection site. Clearance of pDNA from the injection site was slower in VCL-CT01-vaccinated mice compared with PBS-pDNA-vaccinated mice. An integration study was conducted in rabbits to determine whether pDNA integration into the genome of the vaccinated animal contributed to pDNA persistence. Residual pDNA in VCL-CT01 injected rabbit muscle collected 60 days after vaccination (geometric mean of 1085 PCN/microg total DNA) was comparable to that observed in VCL-CT01- injected mouse muscle (geometric mean of 1471 PCN/microg total DNA) collected at the same time point. pDNA integration was not detectable by column agarose gel electrophoresis despite the persistence of pDNA at the injection site 60 days after vaccination. Therefore the risk of genomic integration of hCMV pDNA formulated with poloxamer was considered negligible. PMID- 16218773 TI - Control of viral infectivity by tripartite motif proteins. AB - It is of great interest to understand the molecular details of the pathways that constitute species barriers to viral infection. The tripartite motif protein TRIM5alpha has emerged as an important mediator of species-specific retroviral replication and innate immunity. This review considers the role of TRIM5alpha as an antiviral protein in mammals. The methods used to identify species-specific restriction to retroviral infection, and the identification of TRIM5alpha itself, are outlined. TRIM5alpha mediates an early postentry block to sensitive retroviral infection, usually before viral DNA synthesis. Results from mutational analysis of TRIM5alpha and their contribution to a mechanistic model for TRIM5alpha antiviral activity are discussed. The antiviral role of other TRIM proteins is considered, as is the role of TRIM5alpha cytoplasmic bodies. PMID- 16218776 TI - II. Cationic lipid-formulated plasmid DNA-based Bacillus anthracis vaccine: evaluation of plasmid DNA persistence and integration potential. AB - Several formulated plasmid DNA (pDNA)-based vaccines are being evaluated for safety and efficacy in healthy human subjects. A safety concern for any vaccine that contains genetic material, be it whole organism, live-attenuated, or gene based, is the potential for integration into genomic DNA (gDNA). To address this concern, a preclinical pDNA persistence/integration study was conducted in rabbits to determine the level of pDNA in muscle 2, 28, and 64 days after intramuscular injection of DMRIE:DOPE-formulated pDNAs encoding Bacillus anthracis detoxified LF and PA proteins (VCL-AB01 vaccine). Total DNA was extracted from day 64 muscle tissue and fractionated by column agarose gel electrophoresis (CAGE). Plasmid copy number (PCN) in muscle 64 days after injection (geometric mean, 2808 PCN/microg of total DNA or 150,000 diploid genomes) was determined by quantitative polymerase chain reaction. Analysis of total DNA from five VCLAB01- injected rabbits revealed that two of five samples had no detectable PCN in the high molecular weight fraction after one round of CAGE, two samples had PCN under the lower limit of quantitation, and the remaining sample had 123 PCN/microg. All PCN in the latter sample cleared after an additional round of CAGE. It appears, therefore, that persisting PCN fractionate as low molecular weight material and are most likely not integrated into gDNA. Even if the worst-case assumption is made that the highest PCN found associated with gDNA represented covalently integrated pDNA inserts, the frequency of mutation would still be 500-fold lower than the autosomal spontaneous mutation rate. PMID- 16218777 TI - Amino-terminal fragment of urokinase inhibits tumor cell invasion in vitro and in vivo: respective contribution of the urokinase plasminogen activator receptor dependent or -independent pathway. AB - The urokinase plasminogen activator (uPA) is implicated in both cancer cell invasion and angiogenesis. It can interact with a specific receptor (uPAR) via the epidermal growth factor (EGF)-like domain in the urokinase amino-terminal fragment (ATF) in a species-specific manner. Our previous studies showed that adenovirusmediated delivery of murine ATF (AdmATF) suppressed human tumor growth in mouse models, by inhibiting murine angiogenesis. However, we cannot exclude its putative inhibitory action on human cancer cell invasion through a uPAR independent pathway. To further investigate the mechanisms of ATF, we constructed another adenovirus, AdhmATF, expressing humanized murine ATF (hmATF). hmATF binds to human uPAR but not to murine uPAR. We compared the antagonist effect of both AdmATF and AdhmATF on human and murine cancer cells. In vitro, the supernatant from AdhmATF-infected cells repressed 79% of membrane-associated uPA activity on human MDA-MB-231 cells, whereas that from AdmATF-infected cells repressed 35% of membrane-associated uPA activity. On murine LLC cells, the supernatant from AdhmATF-infected cells inhibited 29% of cell surface uPA activity, whereas that from AdmATF-infected cells inhibited 74% of cell surface uPA activity. Similar results were obtained in a cell invasion assay. In vivo, intratumoral injection of the adenoviruses into LLC tumors on day 24 postinjection induced lower but significant tumor growth suppression by AdhmATF (tumor volume was 1185 +/- 128 mm3), whereas suppression by AdmATF was greater (407 +/- 147 mm3). In the MDA-MB 231 tumor model, on day 52 postinjection, tumor size was 187 +/- 47 mm3 in the AdhmATF-treated group and 468 +/- 65 mm3 in the AdmATF-treated group. The LLC and MDA-MB- 231 cell lines transfected by mATF or hmATF genes showed growth inhibition In vivo equivalent to the results obtained by adenovirus treatment. These results demonstrate the strong anticancer activity of ATF even when its uPAR-binding affinity has been suppressed, and indicate that ATF exerts an antitumor effect via dual mechanisms: essentially through targeting the uPA-uPAR system via the EGF-like domain and partially through targeting a uPAR-independent interaction via the kringle domain. PMID- 16218778 TI - Efficient in vivo xenogeneic retroviral vector-mediated gene transduction into human hepatocytes. AB - We developed a method for efficient retroviral vector-mediated gene transfer into human hepatocytes, using a human hepatocyte-bearing mouse model. Normal human hepatocytes were transplanted into the livers of immunodeficient and liver damaged mice. Donor hepatocytes multiplied and replaced the host hepatocytes, which yielded human hepatocyte-bearing mice (human hepatocyte-chimeric mice). As control cells, rat hepatocytes were similarly transplanted. The replacement level reached 86% at 8 weeks and 100% at 5 weeks posttransplantation of human and rat hepatocytes, respectively. Human and rat hepatocytes in the host liver showed a high bromodeoxyuridine-labeling index during the first 2 weeks posttransplantation. Human- and rat-chimeric mice were injected 7 and 10 days posttransplantation, respectively, with retroviral vectors carrying the beta galactosidase gene and were thereafter injected daily for 20 and 10 days, respectively. The level of beta-galactosidase-positive hepatocytes in the human- and rat-chimeric mice reached 7.1 +/- 1.8% at 8 weeks and 5.3 +/- 0.9% at 5 weeks after transplantation, respectively. The human hepatocyte-chimeric mouse will be useful for testing the ability of vectors to transduce human cells. PMID- 16218779 TI - Characterization of the bystander effect of somatostatin receptor sst2 after in vivo gene transfer into human pancreatic cancer cells. AB - Pancreatic cancer is one of the most aggressive and devastating human malignancies. The present study was conducted to determine whether in vivo sst2 gene transfer into human pancreatic tumors would impair tumor progression, and to characterize sst2 antitumoral bystander mechanisms. sst2 administration, using the synthetic vector PEI, strongly inhibited tumor progression of human pancreatic adenocarcinoma, in vivo. sst2 gene transfer induced intratumoral production of its ligand somatostatin. Disruption of this autocrine loop by RNA interference completely reversed sst2 antitumoral activity. Mice depleted of natural killer (NK) cells did not hamper sst2 tumor growth inhibition. However, microvessel density and vascular endothelial growth factor (VEGF) expression were markedly reduced in sst2-transfected tumors, whereas sst3 somatostatin receptor was upregulated. Depleting somatostatin by RNA interference completely abolished the sst2 inhibitory effect on VEGF expression and tumor angiogenesis, and sst2 induced sst3 expression in peripheral tumor vessels. We conclude that in vivo sst2 gene transfer elicited intratumoral somatostatin production and strongly impaired human pancreatic tumor growth. NK cells were not involved in this antitumoral bystander effect. VEGF and tumor vascularization were identified as novel targets for sst2-mediated antitumoral bystander effect. sst3 somatostatin receptor was upregulated in sst2-transfected tumors. Therefore, in vivo gene delivery of sst2 receptor to target the angiogenic process in pancreatic ductal adenocarcinoma might be a new therapeutic approach for treatment of pancreatic cancer in patients with unresectable disease. PMID- 16218780 TI - Electrophoretic component of electric pulses determines the efficacy of in vivo DNA electrotransfer. AB - Efficient DNA electrotransfer can be achieved with combinations of short high voltage (HV) and long low voltage (LV) pulses that cover two effects of the pulses, namely, target cell electropermeabilization and DNA electrophoresis within the tissue. Because HV and LV can be delivered with a lag up to 3000 sec between them, we considered that it was possible to analyze separately the respective importance of the two types of effects of the electric fields on DNA electrotransfer efficiency. The tibialis cranialis muscles of C57BL/6 mice were injected with plasmid DNA encoding luciferase or green fluorescent protein and then exposed to various combinations of HV and LV pulses. DNA electrotransfer efficacy was determined by measuring luciferase activity in the treated muscles. We found that for effective DNA electrotransfer into skeletal muscles the HV pulse is prerequisite; however, its number and duration do not significantly affect electrotransfer efficacy. DNA electrotransfer efficacy is dependent mainly on the parameters of the LV pulse(s). We report that different LV number, LV individual duration, and LV strength can be used, provided the total duration and field strength result in convenient electrophoretic transport of DNA toward and/or across a permeabilized membrane. PMID- 16218781 TI - Arteriogenic gene therapy in patients with unreconstructable critical limb ischemia: a randomized, placebo-controlled clinical trial of adenovirus 5 delivered fibroblast growth factor-4. AB - The objectives of this study were to assess the safety and potential clinical efficacy of adenovirus-delivered fibroblast growth factor-4 (Ad5FGF-4) by intramuscular injection into patients with critical limb ischemia (CLI). This study was a double-blind, randomized, placebo-controlled study with escalating dose groups of 2.87 x 10(8) to 2.87 x 10(10) viral particles. Thirteen patients with CLI were randomized to receive active drug (n = 10) or placebo (n = 3). Safety evaluations and efficacy parameters (ankle-brachial index, digital subtraction angiograms, magnetic resonance imaging, and scintigraphy) were performed at baseline and for 12 weeks after treatment. Injections of Ad5FGF-4 were generally well tolerated and considered safe. Transfection efficacy at these concentrations may have been limited or local. The small sample size did not allow any firm conclusions regarding clinical efficacy but a trend toward more and slightly larger blood vessels was observed in the angiograms. It is concluded that intramuscular injection of Ad5FGF-4 into CLI patients seemed safe, but transfection efficacy was limited at the assessed doses. Conclusions regarding clinical efficacy are impossible to draw from this small patient cohort. PMID- 16218782 TI - Large-scale production of recombinant viruses by use of a large culture vessel with active gassing. AB - Adenovirus and adeno-associated virus (AAV) vectors are increasingly used for gene transduction experiments. However, to produce a sufficient amount of these vectors for in vivo experiments requires large-capacity tissue culture facilities, which may not be practical in limited laboratory space. We describe here a large-scale method to produce adenovirus and AAV vectors with an active gassing system that uses large culture vessels to process labor- and cost effective infection or transfection in a closed system. Development of this system was based on the infection or transfection of 293 cells on a large scale, using a large culture vessel with a surface area of 6320 cm2. A minipump was connected to the gas inlet of the large vessel, which was placed inside the incubator, so that the incubator atmosphere was circulated through the vessel. When active gassing was employed, the productivity of the adenovirus and AAV vectors significantly increased. This vector production system was achieved by improved CO2 and air exchange and maintenance of pH in the culture medium. Viral production with active gassing is particularly promising, as it can be used with existing incubators and the large culture vessel can readily be converted for use with the active gassing system. PMID- 16218783 TI - Adeno-associated virus inverted terminal repeats improve neuronal transgene expression mediated by baculoviral vectors in rat brain. AB - Baculoviral vectors can transduce neurons in the CNS but mediate only transient expression of transgenes. We have developed a new baculoviral vector in which the inverted terminal repeats (ITRs) of adeno-associated virus are used to flank a luciferase reporter gene cassette harboring a neuron-specific promoter. When tested in rat brain, the new viral vector was able to provide transgene expression for at least 90 days. Immunohistological analysis demonstrated that ITR flanking did not affect the cellular preference of the neuronal promoter in the context of baculovirus. These findings establish an effective way to engineer baculoviral vectors in order to achieve sustained expression of a functional gene for gene therapy for neurodegenerative disorders and physiological studies of neurons. PMID- 16218785 TI - Recommendations of the Gene Therapy Advisory Committee/Committee on safety of medicines working party on retroviruses. May 2005. PMID- 16218784 TI - Product-enhanced reverse transcriptase assay for replication-competent retrovirus and lentivirus detection. AB - The product-enhanced reverse transcriptase (PERT) assay has been used to detect reverse transcriptase (RT) activity associated with retroviruses. Although the PERT assay has been proposed as a method for detection of replication-competent retrovirus (RCR) and lentivirus (RCL), it has not been rigorously compared with existing methods for RCR and RCL detection. We have assessed the PERT assay for detection of RCL and RCR that may contaminate lentiviral and retroviral vectors and compared it with published methods for RCL (p24gag ELISA/gag PCR) and RCR (S+/L-) detection. Our results suggest that the PERT assay is as sensitive as p24gag ELISA and gag PCR for detection of replication-competent HIV-1 in an RCL detection assay. Comparison of detection of replication-competent retroviruses, GALV and RD114, by extended S+/L- and PERT assays indicates that both assays can detect 1 IU of each virus. Our findings suggest that the PERT assay can be used for RCL and RCR testing of a variety of retroviral vectors regardless of the structure, sequence, and envelope of the vectors. PMID- 16218797 TI - Reasons for stopping antiretrovirals used in an initial highly active antiretroviral regimen: increased incidence of stopping due to toxicity or patient/physician choice in patients with hepatitis C coinfection. AB - Low adherence and toxicities among HIV-positive patients starting highly active antiretroviral therapy (HAART) can lead to discontinuation of therapy and treatment failure. Little is known about hepatitis C (HCV) status and discontinuation of HAART. Poisson regression was used to determine factors related to discontinuation of any part of an initial HAART regimen due to treatment failure (TF) or toxicities and patient/physician choice (TOX), and to investigate the relationship between HCV and discontinuation of a HAART regimen in 1198 patients staring HAART after 1999 from the EuroSIDA study. At 1 year after starting HAART, 70% of patients remained on their original regimen, 24% had changed, and 6% were off all treatment. The most frequent reason for discontinuation was toxicities (30.4%). There was no change over time in the proportion of patients discontinuing after stratification by reason for discontinuation (p = 0.18). Of patients 190 stopped at least one antiretroviral drug used in their initial HAART regimen due to toxicities; the toxicity reported did not vary according to HCV status (p = 0.90). Anti-HCV seropositive patients had a higher incidence of discontinuation due to TOX (IRR 1.46, 95% CI 1.13-1.88, p = 0.0042) compared to patients without HCV. Patients with HCV were more likely to discontinue all or part of their HAART regimens due to toxicity or patient/physician choice. Managing adverse events must remain a key intervention in maintaining HAART. There is a need for further studies to describe the relationship between HCV, specific antiretrovirals, and different treatment strategies. PMID- 16218798 TI - Unusual thromboses associated with protein S deficiency in patients with acquired immunodeficiency syndrome: case reports and review of the literature. AB - Recent reports indicate that patients infected with HIV are at increased risk for the development of thrombosis. Among other possibilities, an acquired deficiency of protein S (PS), one of the plasma's natural anticoagulants, might explain this tendency. PS deficiency can be classified in three types depending on the levels of total and free protein (antigenic assays) as well as anticoagulant activity (functional assay). Although the prevalence of inherited PS deficiency is not known because of its rarity, several conditions can lead to acquired forms of the disease. We report two AIDS patients with coexistent type III PS deficiency and thrombosis. Our first patient presented with bilateral chronic leg ulcers and a skin biopsy revealed dermal microthromboses. On laboratory evaluation he had PS deficiency and was started on anticoagulation, but was lost to follow-up. The second patient presented with hepatic vein thrombosis (Budd-Chiari syndrome) and was also PS deficient. On long-term anticoagulation, she experienced resolution of the thrombosis. Neither patient had prior personal or family history of venous thrombosis, nor acquired risk factors such as immobility, acute infection, recent surgery, or hormonal therapy. The literature contains a few reports of skin ulcers and Budd-Chiari syndrome associated with PS deficiency, although none in AIDS patients. While a larger number of studies describe an association between PS deficiency and HIV infection, the causal effect of this deficiency on the thrombophilic tendency in AIDS has not been established. We propose that awareness of the increased risk for thrombosis in HIV infection is important to the understanding of disease pathophysiology and management of these patients. PMID- 16218799 TI - A randomized trial of the efficacy and safety of fenofibrate versus pravastatin in HIV-infected subjects with lipid abnormalities: AIDS Clinical Trials Group Study 5087. AB - There is a paucity of information on the safety and efficacy of lipid-lowering therapy for dyslipidemia associated with human immunodeficiency virus (HIV) and antiretroviral therapy. Our objective was to determine whether fenofibrate and pravastatin were equivalent for the treatment of combined dyslipidemia in HIV as measured by a composite of the National Cholesterol Education Project (NCEP) goals based on absolute values for low-density lipoprotein (LDL), triglycerides (TG), and high-density lipoprotein (HDL) and to compare the safety of these agents through 48 weeks. This was a randomized, open-label trial with subjects assigned to fenofibrate 200 mg (n = 88) or pravastatin 40 mg (n = 86) daily. Subjects who failed to reach the NCEP composite goal on monotherapy by week 12 received both drugs. The composite goal at week 12 was achieved in 1% of fenofibrate and 5% of pravastatin subjects. At week 16, 69/88 subjects on fenofibrate added pravastatin (FP) and 67/86 subjects on pravastatin added fenofibrate (PF). At week 48, 7% FP subjects and 3% PF subjects achieved the composite goal. Median changes in LDL/HDL/TG/non-HDL were -8/+5/-144/+50 and 14/+2/-66/+34 mg/dl in subjects receiving FP and PF, respectively. There were few adverse events and no rhabdomyolysis reported. Combination therapy with fenofibrate and pravastatin for HIV-related dyslipidemia provides substantial improvements in lipid parameters and appears safe, but is unlikely to achieve all NCEP targets for lipid levels. PMID- 16218800 TI - Seminal reservoirs during an HIV type 1 eradication trial. AB - Despite dramatic reduction of the levels of human immunodeficiency virus type I (HIV-1) virions in blood and seminal plasma of infected patients, highly active antiretroviral therapy (HAART) does not eradicate HIV-1. Three patients, with less than 50 copies/ml of plasma viral RNA, were enrolled in this eradication protocol. Didanosine (DDI) and hydroxyurea (HU) were added to their baseline HAART and after a month of therapy, low dose OKT3, followed by a 2-week course of interleukin 2 (IL-2), was administrated. All antiretroviral therapy was then interrupted and the three patients developed viral rebound in the peripheral blood. The V3 loop region of the HIV-1 gp120 from cell-free viral RNA and proviral DNA in blood and seminal compartments was sequenced in one patient. The two major viral isolates in semen cells were macrophage- tropic (R5) and dual tropic (R5X4), and these isolates were also present in the PBMCs. Six months after the viral rebound, we demonstrated a shift toward dual tropism in semen cell-associated HIV-1 proviral DNA, with the first appearance of a T-lymphotropic (X4) provirus solely in this compartment. The virus responsible for the blood plasma viral rebound was never found in the semen microenvironment. This study suggests viral compartmentalization of the semen microenvironment after an intensification and stimulatory HIV-1 eradication protocol, with evidence of viral evolution. PMID- 16218801 TI - In vitro generation of HIV type 1 subtype C isolates resistant to enfuvirtide. AB - Enfuvirtide (ENF) is the first in a new class of antiretroviral agents targeting the fusion process of the viral life cycle. ENF is a synthetic 36-amino acid peptide that binds to the HR-1 region of gp41 preventing fusion of viral and cellular membranes. With the introduction of ENF there are now four classes of antiretrovirals each with distinct and different resistance pathways. Resistance to ENF among subtype B HIV-1 isolates is associated with amino acid changes mainly in the HR-1 region, although other regions of envelope have also been implicated. To determine whether subtype C viruses developed resistance mutations similar to subtype B viruses, 11 subtype C and 4 subtype B viruses were passaged in the presence of increasing concentrations of ENF. The subtype C isolates showed varying levels of replication at 1 microg/ml ENF by day 18, but by day 29 all replicated efficiently at 10 microg/ml ENF. All subtype C isolates showed evidence of genotypic changes in gp41 HR-1 following exposure to ENF that included G36S/E/D, I37T, V38M/A/L/E, N/S42D, N43K, L45R/M, and A50T/V. Three subtype C viruses had compensatory changes in the HR-2 region, which corresponds to the ENF sequence, and two isolates had changes in the V3 region. Mutational patterns among the four subtype B viruses were similar to those for subtype C and those previously published in the literature. These data indicate that in vitro resistance to ENF develops rapidly among HIV-1 subtype C isolates. In general, mutational patterns for subtype C were similar to those described for subtype B, suggesting that the mechanism of action for ENF is similar for HIV-1 subtype B and C isolates. PMID- 16218802 TI - Single-round HIV type 1 neutralization measured by ELISPOT technique in primary human cells. AB - To accurately quantify HIV-1 neutralizing antibody titers in primary human cells, we developed a single round, focus-forming unit (FFU) reduction assay in human peripheral blood mononuclear cells (PBMC). Infected PBMC were enumerated by a reverse ELISPOT technique in which they were incubated under agarose in the presence of a protease inhibitor in anti-p24 antibody-coated microtiter plates. Viral p24, secreted in the immediate vicinity of infected cells and captured by immobilized antibodies, was subsequently stained using gold-labeled anti-p24 antibody and a precipitating silver substrate. The resulting spots were counted visually, without the aid of a microscope, and percent neutralization titers were determined using curve-fitting software. Results of this ELISPOT neutralization assay (ENA) for 15 HIV-positive human specimens were compared with results from a standard PBMC neutralization assay (standard assay) that measured neutralization as a function of p24 concentration by enzyme immunoassay (EIA). The ENA measures FFU reduction of both syncytium-inducing (SI) and non-syncytium-inducing (NSI) primary isolates. Completed assay plates may be retained as a physical record of results or saved as an image using a flat-bed computer scanner. PMID- 16218803 TI - Distinct profile of T cell activation in HIV type 2 compared to HIV type 1 infection: differential mechanism for immunoprotection. AB - The mechanism for the lower rate of disease progression in HIV-2 infection remains undefined. We evaluated T cell activation in a cohort of HIV-infected commercial sex workers in Dakar, Senegal. CD8+ T cell activation was significantly lower in HIV-2- compared to HIV-1-infected volunteers and both groups displayed higher activation levels compared to seronegative individuals. In contrast, CD4+ T cell activation was similar between the HIV-1 and HIV-2 groups and significantly higher compared to the seronegative group. Interestingly, HIV-2-positive volunteers with evidence of Gag-specific CD8+ T cell responses displayed lower CD4+ T cell activation. Our data suggest that the distinct T cell activation profile in HIV-2-positive individuals may reflect on the presence of effective host immune responses in HIV-2 infection. PMID- 16218804 TI - Genotypic analysis of the protease and reverse transcriptase of HIV type 1 subtype C isolates from antiretroviral drug-naive adults in Malawi. AB - The protease (PR) and reverse transcriptase (RT) regions of HIV-1 isolates from 21 antiretroviral (ARV)-naive Malawian adults were sequenced and analyzed to determine the prevalence of drug resistance-associated mutations in this population. Phylogenetic analysis confirmed that all isolates grouped with HIV-1 subtype C, the predominant subtype in Malawi. No major mutations associated with resistance to PR inhibitors (PIs), nucleoside RT inhibitors (NRTIs), or nonnucleoside RT inhibitors (NNRTIs) were found. In contrast, accessory mutations were found in the protease region at positions 10, 20, 36, 63, 77, and 93, and in the RT region at positions 118, 211, and 214. Further studies will be needed to determine the clinical impact of these polymorphisms on viral susceptibility to existing antiretroviral drugs. PMID- 16218805 TI - Determination of gag subtypes of HIV type 1 detected among female sex workers in Calcutta, India. AB - HIV-1 subtyping is important to study the changing scenario of genetic variation. The gag-based heteroduplex mobility assay (gag-HMA) was developed and evaluated as a powerful and reliable technique for identifying the HIV-1 group M subtypes A to H and the circulating recombinant forms (CRFs). To study the subtype distribution of HIV-1 strains from the eastern part of India, we used the gag based HMA, followed by the sequencing and phylogenetic analysis. Blood samples from HIV-1-seropositive female sex workers in Calcutta were subjected to gag-HMA. The most prevalent subtype was found to be the C type, among which the C4 subsubtype was prevalent. However, a number of nontypable C strains were found in gag-HMA. Phylogenetic analysis revealed the discrete nature of the C strains and no monophyletic cluster was noticed. This result might indicate a growing tendency of variations among the HIV-1 type C strains circulating in eastern India. PMID- 16218806 TI - HIV type 1 subtypes in circulation in northern Kenya. AB - The genetic subtypes of HIV-1 circulating in northern Kenya have not been characterized. Here we report the partial sequencing and analysis of samples collected in the years 2003 and 2004 from 72 HIV-1-positive patients in northern Kenya, which borders Ethiopia, Somalia, and Sudan. From the analysis of partial env sequences, it was determined that 50% were subtype A, 39% subtype C, and 11% subtype D. This shows that in the northern border region of Kenya subtypes A and C are the dominant HIV-1 subtypes in circulation. Ethiopia is dominated mainly by HIV-1 subtype C, which incidentally is the dominant subtype in the town of Moyale, which borders Ethiopia. These results show that cross-border movements play an important role in the circulation of subtypes in Northern Kenya. PMID- 16218807 TI - Analysis of the env V3 sequences obtained from patients with HIV type 1 infection treated with the immune modulant agent tucaresol. AB - Tucaresol, a Schiff base-forming compound that is shown to enhance cytotoxic T cell responses and the production of type 1 cytokines, represents a potentially useful adjuvant factor for treating HIV-1 infection. We studied the effect of tucaresol on V3 sequences within the HIV-1 env region derived from patients with different virologic and immunologic features who were enrolled in a phase I/II randomized clinical trial. The sequence analysis of the env V3 region of the viruses at baseline has confirmed a genotypic pattern similar to a macrophagotropic virus model; we analyzed the follow-up sequences at week 16 of the protocol and did not observe any difference in the tropism determinants within the third variable fragment of the env region. The administration of tucaresol did not accelerate env V3 evolution thus preventing modifications of HIV-1 tropism over time. PMID- 16218808 TI - Chimpanzee DC-SIGN alleles predict the existence of A and B isoforms, but do not support a role for resistance to HIV infection. AB - We report here the gene sequence for DC-SIGN (CD209) from chimpanzees. DC-SIGN is a C-type lectin expressed by dendritic cells. It is involved in DC-T cell interactions as well as in HIV-1 and SIV transmission. We have cloned two new alleles for chimpanzee DC-SIGN. The coding sequences are highly homologous to the two previously described chimpanzee alleles. We confirm the existence of a polymorphism within the repeat region of DC-SIGN. In humans polymorphisms in the repeat region have been associated with resistance to HIV infection. However, we have not been able to correlate the number of repeats with susceptibility of chimpanzees to HIV infection. The actual impact of DC-SIGN variability in HIV infection therefore remains to be determined. PMID- 16218809 TI - The HIV type 1 epidemic in Belarus: predominance of Eastern European subtype A strains and circulation of subtype B viruses. AB - To study the molecular epidemiology of HIV-1 in Belarus, where the rapid spread of HIV-1 has been registered since 1996, we obtained HIV-1 sequences from 30 individuals living in five cities in both the main geographic areas of the epidemic (Gomel and Minsk regions) and territories where spreading of the epidemic remains limited (Grodno region). Analysis of env V3 and gag p17/p24 sequences demonstrated that infections in all 12 injecting drug users and 14 of 18 individuals infected through sexual contacts were caused by subtype A viruses that are specific for the epidemic in the former Soviet Union (IDU-A viruses), while the remaining four infections were caused by phylogenetically unrelated to each other subtype B viruses. Extrapolation of these results to the total population of HIV-1-infected individuals in Belarus allowed us to estimate that IDU-A viruses account for nearly 95% of HIV-1 infections in Belarus. PMID- 16218811 TI - Differentiation potential. PMID- 16218812 TI - Development of reproductive biotechnologies in domestic animals from artificial insemination to cloning: a perspective. AB - There has been a remarkable increase in knowledge resulting in the application of reproductive biotechnologies in animals, with profound implications for human beings. The major accomplishments in domestic animals, particularly with cattle, are reviewed here. An attempt is made to examine these, in perspective, to indicate the steps by which progress was made, and to appreciate that the explosion of new findings today would not have been possible without the careful studies of yesteryears. PMID- 16218813 TI - Optimization strategies for production of mammalian embryos by nuclear transfer. AB - In order to optimize each of the individual steps in the nuclear transfer procedure, we report alternative protocols useful for producing recipient cytoplasts and for improving the success rate of nuclear transfer embryos in cattle, rhesus monkey, and hamster. Vital labeling of maternal chromatin/spindle is accomplished by long wavelength fluorochromes Sybr14 and rhodamine labeled tubulin allowing constant monitoring and verification during enucleation. The use of Chinese hamster ovary (CHO) donor cells expressing the viral influenza hemagglutinin fusion protein (HA-300a+), to adhere and induce fusion between the donor cells and enucleated cow, rhesus and hamster oocytes was examined. Cell surface hemagglutinin was activated with trypsin prior to nuclear transfer and fusion was induced by a short incubation of a newly created nuclear transfer couplet at pH 5.2 at room temperature. Donor cell cytoplasm was dynamically labeled with CMFDA, or further transfected with the green fluorescence protein (GFP) gene, so that fusion could be directly monitored using live imaging. High rates of fusion were observed between CHO donor cells and hamster (100%), rhesus (100%), and cow recipient cytoplasts (81.6%). Live imaging during fusion revealed rapid intermixing of cytoplasmic components between a recipient and a donor cell. Prelabeled donor cytoplasmic components were uniformly distributed throughout the recipient cytoplast, within minutes of fusion, while the newly introduced nucleus remained at the periphery. The fusion process did not induce activation as evidenced by unchanged distribution and density of cortical granules in the recipient cytoplasts. After artificial activation, the nuclear transfer embryos created in this manner were capable of completing several embryonic cell divisions. These procedures hold promise for enhancing the efficiency of nuclear transfer in mammals of importance for biomedical research, agriculture, biotechnology, and preserving unique, rare, and endangered species. PMID- 16218817 TI - Cloning LiteratureWatch 1997-1999. PMID- 16218814 TI - Comparison of oocyte-activating agents for mouse cloning. AB - Since somatic cell components are unable to activate oocytes following injection or fusion, enucleated oocytes receiving adult somatic cells during the cloning process must be activated artificially for their development. We compared the efficiency of four types of oocyte-activating agents: strontium, ethanol, single electric pulse, and spermatozoa. Although strontium was the best in supporting preimplantation development of reconstructed mouse oocytes, there was no significant difference among the four agents with respect to the rate of postimplantation embryo development and the birth of live offspring. PMID- 16218815 TI - Mammalian leukocytes contain all the genetic information necessary for the development of a new individual. AB - We have used leukocytes and oocytes from commercially slaughtered animals to clone a progeny tested Brown Swiss bull. Mononuclear cells were separated from the heparinized blood of the donor male on a Histopaque gradient and cryopreserved. The nuclei of thawed leukocytes were directly microinjected into enucleated Holstein Friesian oocytes that were subsequently activated. Development to morula was 23% and to blastocysts was 17%. Some of the cloned compacting morulae were subjected to a second round of nucleus transfer by fusion of individual blastomeres to enucleated oocytes. Development of these second generation embryos to the blastocyst stage was 19%. Following embryo transfer of 50 blastocysts to 50 recipient heifers (31 from first generation and 19 from second generation), 28 pregnancies were established as evidenced by fetal heartbeat at 35 days. A high proportion of the pregnancies established were lost by day 45. One fetus from a second generation embryo developed to term. The phenotype (Brown Swiss) and DNA analysis (11 microsatellites on 11 different chromosomes) of the resultant normal healthy calf confirmed its identity to the donor sire. The ability to clone animals from hematopoietic cells that can be easily collected and cryopreserved from any donor irrespective of species, age, or sex has important implications for the preservation of genetic resources from a wide variety of animals in the animal breeding and artificial insemination industries and for human medicine. PMID- 16218819 TI - Somatic cell nuclear transfer in mammals: progress and applications. AB - Somatic nuclear transfer has been performed with frogs since the early 1960s, yet it has proved impossible to generate an adult frog using an adult cell as nuclear donor. After some initial skepticism, the birth of sheep, cows, goats, and mice using this technique with fetal or adult cell donors is now established fact. The success with adult mammalian cell donors extends the historic work in frogs by attesting to the totipotency of nuclei in at least some adult, differentiated cell types. Because the technique offers a developmental read out of the totality of genetic and molecular lifetime changes accumulated by the nucleus of a single somatic cell, basic research applications are seen in the fields of ageing, cancer, X chromosome inactivation, and imprinting. The prospect of a method for gene targeting in livestock holds particular promise for commercial applications; whilst for humans, the use of nuclear transfer to provide diverse populations of customized stem cells for therapeutic purposes presents a tantalizing future goal. PMID- 16218820 TI - The effect of donor cell serum starvation and oocyte activation compounds on the development of somatic cell cloned embryos. AB - Two experiments, one comparing nuclear transfer (NT) embryo activation compounds, the other donor cell treatments, were conducted with a goal of identifying factors that improve the in vitro development of cloned bovine embryos. In experiment 1, 539 NT embryos were produced by combining serum starved bovine fetal fibroblasts with enucleated in vitro matured oocytes, activated with ionomycin, then randomly allocated to be incubated for 4 hours in either Butyrolactone-I (BL-I) or 6-dimethylaminopurine (DMAP). There was no significant difference in development to blastocyst or compact morula of fused embryos at Day 6.5 between BL-I and DMAP activated embryos (22.4% vs. 20.2%; p = 0.18). Karyotyping of 20 blastocysts and compact morula from each group determined that 65% of BL1 and 63% of DMAP embryos were diploid with the remainder mixoploid (2n + 4n). In Experiment 2, the development of 389 NT embryos reconstructed from either serum starved or serum fed fetal fibroblasts was assessed. More Day 7 blastocysts and compact morula developed in the serum starved group (34.5% vs. 18.8%; p = 0.008). To verify the viability of BL-I activated embryos, 10 blastocytes from experiment 2 were transferred into 4 recipient cows. Two morphologically normal fetuses, genetically identical to the original fetal cell line, were surgically recovered at day 45 of gestation. In summary, serum starvation of bovine fetal fibroblasts prior to NT significantly improved development to blastocyst. Additionally, we have shown that BL-I is a novel alternative compound for use in combination with ionomycin to activate NT embryos. PMID- 16218821 TI - Parthenogenetic activation of porcine oocytes by electric pulse and/or butyrolactone I treatment. AB - The goal of the present research was to study the parthenogenetic activation of porcine oocytes following treatment with the specific cyclin-dependent kinase inhibitor butyrolactone I (BL I). In Experiment I, the effective dose of BL I was determined by the rates of the subsequent pronuclear formation in oocytes after the activation. In Experiment II, BL I was further tested alone or in combination with an electric pulse. The efficiency of the various treatments to induce activation and parthenogenetic development was examined. In Experiment III parthenogenetic development of activated oocytes in two different media was compared. Cleavage and blastocyst developmental rates were examined, and number of cells in the blastocysts was determined. Our results indicate that, in pig, the optimal activation dose for BL I was 150 microM; a combined electrical and BL I treatment resulted in superior cleavage rates compared to an electric pulse, 150 microM of BL I, or 200 microM of BL I alone (74%, 60%, 41%, and 42%, respectively; P < 0.05); and the rate of parthenogenetic development of activated oocytes to the blastocyst stage in mNCSU37 medium was significantly higher than that in Whitten's medium (59% vs. 5%, P < 0.05) and the resulting day-6 blastocysts had higher cell numbers (35.5 +/- 14.1 vs. 19.5 +/- 2.5). This activation protocol might be useful in porcine nuclear transfer experiments and for the generation of parthenogenetic fetuses. PMID- 16218822 TI - Immunomagnetic isolation of primordial germ cells and the establishment of embryonic germ cell lines in the mouse. AB - The stage-specific embryonic antigen 1 (SSEA-1) is a cell marker of primordial germ cells (PGCs). In the present study, it is shown that isolation and purification of PGCs from 8.5-11.5 days post coitum (dpc) embryos can be achieved by a immunomagnetic cell sorting method using SSEA-1 antibody-conjugated magnetic beads, and then the sorted PGCs can be used for long-term culture under strict culture conditions to derive embryonic germ (EG) cell lines. Five independent EG cell lines with male karyotypes have been established. They show both a strong alkaline phosphatase activity and expression of the SSEA-1 antigen, and are karyotypically stable with a modal number of chromosomes in more than 80% of the cells. One of the EG cell lines from 8.5-dpc embryos produced chimeras after injections of the cells into 8-cell host embryos. These procedures could provide a useful and simple method for isolation of undifferentiated cells from a heterogeneous cell population and for establishment of embryo-derived stem cell lines. PMID- 16218823 TI - Draft National Institutes of Health guidelines for research involving human pluripotent stem cells (December 1999). AB - The National Institutes of Health (NIH) is requesting public comment on a document entitled "Draft National Institutes of Health guidelines for research involving human pluripotent stem cells (December 1999)". The purpose of these draft guidelines is to recommend procedures to help ensure that NIH-funded research in this area is conducted in an ethical and legal manner. The NIH will not fund research using human pluripotent stem cells until final guidelines are published in the Federal Register and an oversight process is in place. PMID- 16218826 TI - Nuclear equivalence, nuclear transfer, and the cell cycle. AB - The last 20 years have seen the development of techniques for the production of mammals by nuclear transfer. Originally limited to the swapping of pronuclei and the use of early cleavage-stage embryos as nuclear donors, nuclear transfer came of age in 1995 with the birth of 2 Welsh Mountain lambs, Megan and Morag, that were produced using cultured differentiated cells as donors of genetic material. In 1996, Dolly was the first animal to be produced using the genetic material from an adult-derived somatic cell. The techniques used in the production of these animals have now been reproduced in both sheep and cattle, and as predicted, successful development has been obtained using donor cells taken directly ex vivo. This article reviews the current status of mammalian nuclear transfer and the biological background to these successes. PMID- 16218828 TI - Transgenic animals: current and alternative strategies. AB - Transgenic animal technology is one of the most fascinating technologies developed in the last two decades. It allows us to address questions in life sciences that no other methods have achieved. The impact on biomedical and biological research, as well as commercial interests are overwhelming. The questions accompanying this fast growing technology and its diversified applications attract the attention from a variety of entities. Still, one of the most fundamental problems remaining is the search for an efficient and reliable gene delivery system for creating transgenic animals. The traditional method of pronuclear microinjection has displayed great variability in success among species. While an acceptable efficiency in the production of transgenic mice has been attained, the relative low efficiency (<1%) in creating transgenic livestock has become one of the barriers for its application. In the past decades, improvements in producing transgenic livestock have made a slow progression, however, the recent advancement in cloning technology and the ability to create transgenic livestock in a highly efficient manner, have opened the gate to a new era in transgenic technology. Discoveries of new gene delivery systems have created an enthusiastic atmosphere that has made this technology so unique. This review focuses on gene delivery strategies as well as various approaches that may assist the advancement of transgenic efficiency in large animals. PMID- 16218827 TI - Cell cycle analysis of cultured porcine mammary cells. AB - One of the major points of debate in determining the effectiveness of nuclear transfer technology has been the phase of the cell cycle of the donor cell at the time of nuclear transfer. Here, a primary mammary cell line has been isolated and various treatments for synchronization of the cell cycle have been tested. The cells were then simultaneously stained for DNA content and protein content and the percentages of cells in G1, G0, S, and G2 + M were estimated. In the first experiment, cells were either cycling, grown to confluence, or serum-starved for 5 days. Serum starvation increased (p < 0.05) the percentage of cells in G0 compared to confluent or cycling cells from 3% to 8% to 22%. By using forward scatter to determine the size of the cells it was determined that if small cells (7-15 microm) were selected from the serum-starved group 43.9% will be in G(0) as compared to 4.5% of cycling cells and 9.9% of confluent cells. Dimethyl sulfoxide (DMSO) treatment (0%, 0.5%, or 1.0%) for 72 hours (shown to synchronize some cell types in G0) had no effect on the percentage of cells in G0, G1, S, or G2 + M. Treatment with mimosine (0 microM, 0.4 microM, 0.8 microM or 1.2 microM), a compound that should synchronize the cells in G1, increased (p < 0.05) the percentage of cells in G1 from 66.7% (0 microM mimosine) to 79.0% to 82.0%. Finally, treatment with colchicine for 24 hours (shown to synchronize some cell types in G2 + M) increased (p < 0.05) the percentage of cells in G2 + M (0 microM colchicine) from 13.3% to 27.2% to 31.6%. It is concluded that many cell cycle synchronization techniques are effective in porcine mammary cell lines, but none of the techniques are 100% effective. Such results should help elucidate the mechanisms involved in nuclear transfer. PMID- 16218829 TI - Establishment of pregnancy after the transfer of nuclear transfer embryos produced from the fusion of argali (Ovis ammon) nuclei into domestic sheep (Ovis aries) enucleated oocytes. AB - Cloning mammalian species from cell lines of adult animals has been demonstrated. Aside from its importance for cloning multiple copies of genetically valuable livestock, cloning now has the potential to salvage endangered or even extinct species. The aim of this study was to investigate the effect of the bovine and domestic (Ovis aries) ovine oocyte cytoplasm on the nucleus of an established cell line from an endangered argali wild sheep (Ovis ammon) after nuclear transplantation. A fibroblast cell line was established from skin biopsies from an adult argali ram from the People's Republic of China. Early karyotype analysis of cells between 3-6 passages revealed a normal diploid chromosome number of 56. The argali karyotype consisted of 2 pairs of biarmed and 25 pairs of acrocentric autosomes, a large acrocentric and minute biarmed Y. Bovine ovaries were collected from a local abattoir, oocytes aspirated, and immediately placed in maturation medium consisting of M-199 containing 10% fetal bovine serum, 100 IU/mL penicillin, 100 microg/mL streptomycin, 0.5 microg/mL follicle-stimulating hormone (FSH), 5.0 microg/mL luetinizing hormone (LH) and 1.0 microg/mL estradiol. Ovine (O. aries) oocytes were collected at surgery 25 hours postonset of estrus from the oviducts of superovulated donor animals. All cultures were carried out at 39 degrees C in a humidified atmosphere of 5% CO2 and air. In vitro matured MII bovine oocytes were enucleated 16-20 hours after onset of maturation and ovine oocytes within 2-3 hours after collection. Enucleation was confirmed using Hoechst 33342 and UV light. The donor argali cells were synchronized in G0-G1 phase by culturing in Dulbecco's modified Eagle's medium (DMEM) plus 0.5% fetal bovine serum for 5-10 days. Fusion of nuclear donor cell to an enucleated oocyte (cytoplast) to produce nuclear transfer (NT) embryos was induced by 2 electric pulses of 1.4 kV/cm for 30 microsc. Fused NT embryos were activated after 24 hours of maturation by exposure to ionomycin (5 microM, 4 minutes) followed by incubation in 6-dimethylaminopurine (0.2 mM, 4 hours) and cultured in microdrops of CR1aa medium. From a total of 166 constructed nuclear donor cell-bovine cytoplasm NT couples, 128 (77%) successfully fused, 100 (78%) developed to 8-16 cell stage, and 2 (1.56%) developed to the blastocyst stage. The presence of argali nuclei in 8-16 cell stage embryo clones was confirmed after observation of Hoechst 33342 stained embryos under UV light and chromosome analysis of metaphase spreads from blastomeres. A total of 127 constructed nuclear donor cell-ovine cytoplasm NT couples were produced, 101 (80%) successfully fused, 81 (80% of fused) developed to the 16- to 32-cell stage. A total of 28 hybrid (argali-sheep) and 21 sheep-sheep NT embryos were transferred into 6 recipients and 4 recipients, respectively. Two of these recipients, 1 carrying argali-sheep and 1 sheep-sheep, were confirmed pregnant at 49 days by ultrasound, but both pregnancies terminated by 59 days. The results of this study demonstrate the possibility of using xenogenic oocytes to produce early-stage embryos and pregnancies from an established fibroblast cell line of an endangered species. PMID- 16218830 TI - Development of pig embryos by nuclear transfer of cultured fibroblast cells. AB - Pig fibroblast cells were transferred to enucleated oocytes by micromanipulation and electrofusion. The donor cells used for nuclear transfer were synchronized in presumptive G0 by serum starvation. In the first experiment, nuclear transfer was performed with fibroblasts that had either a smooth or a rough surface. A significant difference (p < 0.05) in the percentage of chromosome condensation (39.5%, 15/38 and 16.6%, 5/30) and nuclear formation (36.8%, 14/38 and 16.3%, 8/49) was found between the reconstructed embryos derived from the cells with smooth surface and with rough surfaces, respectively. The percentage of chromosome condensation (42.5%, 17/40 and 19.6%, 11/56) and nuclear formation (38.3%, 23/60 and 18.8%, 9/48) were higher (p < 0.05) in reconstructed embryos derived from small (15 microm) donor cells compared to large donor cells (20 microm), respectively. The percentage nuclei at 3 different time points (3, 6, and 9 hours in culture medium) was higher (p = 0.003) in the reconstructed embryos activated by thimerosal and dithiothreitol (20%, 36%, and 41.3%) compared to those without activation treatment (0%, 11.8%, and 22.2%). In addition, there was an increased percentage with nuclei as the time in culture increased from 3 to 9 hours (p = 0.029). The percentages of chromosome condensation (34.6%; 9/26) and nuclear formation (33.3%; 9/27) in nuclear transfer embryos were similar. The rate of blastocysts/morulae development (14.0%; 6/43) was low. However, 2 cavitated embryos (presumptive blastocysts) with 14 and 11 nuclei and 1 morula with 8 nuclei were obtained. This together with the above evidence indicate that the nuclei from pig fibroblast cells can be partially reprogrammed, which suggests that transfer of nuclei from fibroblast cells to in vitro matured oocytes resulting in production of identical or genetically altered pigs may be possible. PMID- 16218832 TI - Editorial: successes and failures in cloning. PMID- 16218831 TI - Reprogramming of fibroblast nuclei after transfer into bovine oocytes. AB - Recent landmark achievements in animal cloning have demonstrated that the events of cell differentiation can, in principle, be reversed. This reversal necessarily requires large-scale genetic reprogramming, of which little is known. In the present study we characterized the extent to which blastocyst stage-specific mRNA expression would be conserved in bovine embryos produced by nuclear transfer (NT) using fetal fibroblasts as nuclei donors (FF NT). The mRNA pool of FF NT embryos was compared with that of NT embryos reconstructed from embryonic blastomeres (Emb NT), with embryos produced under in vivo or in vitro conditions, and finally with fibroblast cells. Embryo/cell-specific mRNA pools were contrasted using differential display methodology. Random oligonucleotide primer pair combinations were used to subfractionate mRNA populations and represent individual mRNAs as copy DNA (cDNA) bands ranging in size from 100 to 800 base pairs. Regardless of whether bovine blastocysts developed in vivo or in vitro, or were derived after nuclear transplantation with embryonic blastomeres or fetal fibroblasts, their mRNA profile was highly conserved and distinct from that of fetal fibroblast cells. There was approximately 95% conservation in cDNA banding patterns between FF NT, Emb NT, and in vivo derived blastocysts, when compared with in vitro derived blastocysts. In contrast, the cDNA banding in fibroblasts was only 67% conserved with in vitro derived blastocysts (p < 0.0001), indicating that dramatic changes in gene transcription are induced by nuclear transplantation. After nuclear transplantation, gene expression in fetal fibroblasts is reprogrammed so to mimic that of preimplantation embryo development. Future characterization of these changes will be invaluable for the identification of suitable cell types to serve as nuclear donors for embryo reconstruction and provide information that can be used to improve the efficiency of cloning animals by nuclear transplantation. PMID- 16218833 TI - Use of embryonic and somatic cells for production of transgenic domestic animals. AB - In contrast to the highly developed genetic modification systems available for manipulating the mouse genome, at this time only simple gain of function modifications can be undertaken in domestic species. Clearly, the greatest barrier to gene targeting in domestic species has been the unavailability of cell lines that can be modified in vitro and still be used to generate a living organism. In the mouse, the embryonic stem (ES) cells and embryonic germ (EG) cells have fulfilled that role. While the nuclear transfer procedures have solved this problem in sheep and cattle, in swine ES and EG cells are still needed. In addition, targeting in domestic species is affected by the need to develop targeting constructs containing isogenic DNA regions. As a result, it is necessary to isolate the gene of interest, sequence required regions, and develop isogenic targeting constructs by technologies such as long-range PCR. On the positive side, enrichment protocols developed in the mouse can be applied to domestic species, thus facilitating the identification of correctly modified cell lines. Hence, progress in mammalian cloning, the development of EG cell lines, and advances in gene targeting presently allows the introduction of precise genetic modifications into the domestic animal genome. PMID- 16218834 TI - Initiation and organization of events during the first cell cycle in mammals: applications in cloning. AB - The technology of cloning involves transplanting a diploid nucleus into a mature oocyte cytoplast. The cytoplast is then activated to initiate the first cell cycle of development as a nuclear transplant embryo. Initiation and regulation of events during the first cell cycle are, therefore, critical for proper reprogramming of the donor nucleus and development as a cloned embryo. Activation is normally induced by the sperm and is mediated by a series of intracellular free calcium ([Ca(2+)](i)) oscillations that last for several hours. Although it is not known precisely how the sperm induces activation, current evidence favors the delivery, by the sperm, of a soluble protein factor that causes the production of IP3. IP3 acts to open a Ca(2+) channel in the endoplasmic reticulum and release Ca(2+) into the cytosol. A variety of methods have been used to duplicate or replace the sperm-induced [Ca(2+)](i) increase to cause activation in nuclear transplant embryos. It has been found that treatments that cause a single transient [Ca(2+)](i) activate some oocytes with the level of activation increasing as the oocyte ages. Attempts have been made to extend the period of time over which [Ca(2+)](i) oscillations occur. This has been successful in increasing activation rates of less mature oocytes but the techniques are still cumbersome. An alternative method, that has been very successful, is the combination of a treatment that elevates [Ca(2+)](i) and a treatment that maintains low levels of maturation promoting factor for several hours after the initial [Ca(2+)](i) elevation. The sperm also contributes the centrosome that organizes microtubules during the first cell cycle. One current hypothesis for regulation of sperm centrosomal activity consists of a dephosphorylation of sperm connecting piece proteins following sperm entry into the oocyte and activation of the oocyte. Dephosphorylation of these proteins results in the disassembly of the connecting piece and assembly of a functional centrosome. In nuclear transfer, centrosomal components are contributed by the donor cell. If the cell is fused to the cytoplast before centriole replication then a single aster forms. If the cell is fused after centriole replication then two asters form. In either case and even in parthenogenetic oocytes, which do not have centrioles, the first cell cycle progresses to metaphase. However, progress is slow and some defects are observed in the assembly of chromosomes into a metaphase plate. PMID- 16218835 TI - Parthenogenetic activation of porcine oocytes after nuclear transfer. AB - Mature porcine oocytes are arrested at metaphase II of meiosis. At fertilization, like all mammalian oocytes they exhibit a low frequency Ca(2+) oscillation lasting several hours. This oscillation is thought to be the signal that triggers resumption of meiosis and activates the developmental program of the oocyte. The signal transduction mechanism of the sperm-induced Ca(2+) signal is not known in detail, and attempts to generate the oscillation artificially have met with little success. Nevertheless, artificial activation of the oocyte is a crucial step during nuclear transfer. Methods are available to induce a transient elevation in the intracellular free Ca(2+) concentration to surpass the meiotic arrest and induce development of the constructed embryo. Further studies concentrating on the mechanism of Ca(2+) signaling during fertilization will help to improve the efficiency of the procedures used for parthenogenetic activation of the oocyte. PMID- 16218836 TI - Parthenogenetic activation of mouse oocytes by exposure to strontium as a source of cytoplasts for nuclear transfer. AB - Cell-cycle phase of the donor and recipient cells at the moment of nuclear transfer influences subsequent development of the reconstituted embryo. In order to study this effect, the precise cell-cycle phase of the recipient oocyte at the time of fusion must be known and this depends on reliable activation of oocytes in a protocol that has a low incidence of spontaneous activation. Mouse oocytes recovered before (8-10 hours post-human chorionic gonadotropin [hCG]) and after ovulation (14 and 18 hours post-hCG) were exposed to strontium ions in calcium magnesium-free M16 culture medium. The effect on development of haploid parthenotes of post-hCG age of the oocyte, the duration of exposure, and strontium concentration in the medium was determined. These experiments established a reliable method of parthogenetic activation of recently ovulated mouse oocytes, involving the culture of oocytes for 60 minutes in 25 mM strontium in a calcium magnesium-free M16 medium. This method of activation was also able to induce activation of preovulatory oocytes after a preincubation period in vitro. Only a low incidence of spontaneous activation was observed if oocytes were recovered before or immediately after ovulation (14 hours after hCG). PMID- 16218837 TI - Analysis of telomere length in Dolly, a sheep derived by nuclear transfer. AB - We have used a (TTAGGG) oligonucleotide probe to demonstrate that ovine telomeres are composed of (TTAGGG) repeat arrays and to compare the terminal restriction fragment lengths of sheep derived by natural mating and nuclear transfer. Here we show that ovine somatic telomeres decrease in length with age, and that Dolly, derived by the transfer of 6-year-old adult somatic nucleus, exhibits diminished terminal restriction fragment lengths. The decrease is consistent with the age of the donor tissue and telomere erosion during in vitro culture. Nuclear transfer does not restore telomere lengths. Dolly otherwise appears physiologically and phenotypically normal for her breed and age. We further report on apparent telomere lengthening in sheep, occurring during the first year in naturally derived lambs. PMID- 16218840 TI - Corrections. PMID- 16218841 TI - Manipulating the human embryo: cell cycle checkpoint controls. AB - Micromanipulation techniques are widely used in assisted human reproduction and it is logical to assume that successes with recent animal cloning will invariably raise the question of human cloning along with its related ethical problems. However, it is often overlooked that even in animals many complications are still associated with this technique. The purpose of our article is to highlight and discuss some of these problems in the context of the eventual use of nuclear and/or cytoplasmic transfer techniques in assisted human reproduction. PMID- 16218843 TI - Expression of GFP in nuclear transplants generated by transplantation of embryonic cell nuclei from GFP-transgenic fish into nonenucleated eggs of medaka, Oryzias latipes. AB - In order to investigate whether foreign genes can be used as genetic markers of donor nuclei in fish nuclear transplantation, expression of the GFP gene derived from donor nuclei was examined in nuclear transplants in medaka (Oryzias latipes). Embryonic nuclei were obtained from blastula embryos produced by crossing of transgenic fish of the wild-type strain heterozygous for the GFP gene with nontransgenic ones or by mutual crossing between transgenic fish. The GFP gene was driven by the promoter of the medaka elongation factor gene, EF-1alpha A, which is known to induce GFP expression in many tissues except for the muscle in the transgenic fish. The nuclei were transplanted into nonenucleated unfertilized eggs of the orange-red strain. Adult nuclear transplants were successfully obtained at the rate of about 2% of the operated eggs. They were triploid and had no reproductive potential. The GFP gene was expressed in embryos, fry, and adults of nuclear transplants in a pattern similar to that in the transgenic fish. These results indicate that GFP is useful as a foreign genetic marker of donor nuclei in fish nuclear transplantation. PMID- 16218842 TI - Blastomeres from somatic cell nuclear transfer embryos are not allocated randomly in chimeric blastocysts. AB - A marker has been developed to allow detection of blastomeres that originate from embryos produced by nuclear transfer (NT) of genetically engineered fetal fibroblasts. A plasmid (phEFnGFP) was constructed with a G418 resistance cassette for selection in fibroblasts and a nuclear localized green fluorescent protein (nGFP) expression cassette that expresses in every cell of day-6, -7, and -8 bovine embryos. This construct was utilized to follow the blastomere distribution in aggregation chimeras produced from fertilized embryos (in vitro produced, IVP) or parthenotes and NT embryos. Fluorescent and nonfluorescent NT embryos were aggregated early on day 4 and evaluated on day 8. Nuclei of blastomeres that carried the transgene were fluorescent under both UV epifluorescence (Hoechst 33342) and blue epifluorescence (nGFP). There was no bias in the distribution of green fluorescent blastomeres in the inner cell mass (ICM) or trophectoderm in NT<>NT chimeras. However, there was a strong bias for NT blastomeres to populate the ICM when aggregated with IVP embryos or parthenotes. There was also a strong bias against NT blastomeres in the trophectoderm when aggregated to IVP embryos. However, the bias against NT blastomeres in the trophectoderm was significantly less (p < 0.05) when aggregated with parthenotes as compared to aggregation with IVP embryos. In NT<>NT aggregates, no chimeric embryos were produced that had an ICM composed of blastomeres from a single origin. However, in NT<>Parthenote aggregates, 67% of the blastocysts had an ICM composed exclusively of NT origin. The remaining blastocysts ranged from 0% to 83% of the ICM that expressed nGFP. Similarly, in NT<>IVP aggregates 50% of the blastocysts had an ICM composed exclusively of NT origin. The remaining blastocysts ranged from 19% to 71% of the ICM being of NT origin. We conclude that production of divaricated chimeras from NT origin is feasible. Other applications of this technology are discussed. PMID- 16218844 TI - A simplified method for the reconstruction of fully competent mouse zygotes from adult somatic donor nuclei. AB - Until now, full-term development of mouse embryos reconstructed from somatic nuclei has been convincingly achieved only when a piezoimpact pipette drive unit is used for the injection of a donor nucleus into an enucleated recipient oocyte. Here we describe a simplified method for mouse cloning that requires neither electrofusion nor a piezo device. Efficient rates of enucleation can be achieved without staining the chromosomes of the recipient oocyte and high survival rates are obtained after direct injection of the donor nucleus. Although a low proportion of reconstructed embryos could implant after their transfer into the oviducts of foster mothers (less than 5%), we show that at least some of them can develop into normal young. PMID- 16218845 TI - Cloned piglets born after nuclear transplantation of embryonic blastomeres into porcine oocytes matured in vivo. AB - Nuclear transplantation in the pig is more difficult than in other domestic animals and only one embryonic nuclear transplantation (NT) pig has been born to date. In this study, reconstituted porcine embryos were produced by electrofusion of blastomeres from in vivo four-cell embryos to enucleated in vivo or in vitro matured (IVM) oocytes. Nuclear transfer using cumulus cells as nuclear donors was also conducted. When blastomeres were used as donors, the electrofusion rate was significantly higher in oocytes matured in vivo (91.5%) than in those matured in vitro (66.1%) (p < 0.01). After fusion, the NT embryos reconstituted from in vivo matured oocytes developed to blastocysts at a rate of 10.3% after culture in rabbit oviducts for up to 5 days, while only 5.9% of the NT embryos reconstructed from in vitro matured oocytes developed to blastocyst stage. Electrofusion rate of cumulus cell nuclei with enucleated IVM oocytes was lower (47.6%) and only 1.5% (2/136) of the reconstituted eggs developed in vitro to morula stage, and 1.9% developed to blastocysts when cultured in the ligated rabbit oviducts. Transfer of 94 embryos reconstructed by blastomere NT with in vivo matured oocytes to five synchronous recipients resulted in the birth of two cloned piglets. No piglet was born following transfer to two recipients of embryos (n = 39) derived from NT with in vitro matured oocytes. The results demonstrate that in vivo matured oocytes are better recipients than those matured in vitro for pig cloning. PMID- 16218846 TI - The genetically engineering and cloning animals symposium. PMID- 16218847 TI - Changes in the structure of nuclei after transfer to oocytes. AB - Nuclear transfer and the potential for cloning animals have refocused attention on the oocyte. This focus is not limited to the use of the oocyte as a recipient in nuclear transfer procedures, but more broadly in terms of what factors are present in the oocyte that are responsible for establishing the developmental pattern of RNA synthesis and subsequent protein production. Deviations in the pattern of RNA synthesis can result in abortions, as well as abnormalities at birth. This paper will focus on the changes to nuclear structure that result from transfer to the cytoplasm of an oocyte, as well as some of the changes in the patterns of RNA synthesis that have been described. PMID- 16218848 TI - Cloning adult animals - what is the genetic age of the clones? AB - The rapid progress in cloning research along with its many ramifications will soon have a significant beneficial impact on basic research, agriculture, and biomedicine. However, for the nuclear transfer technology to reach its fullest potential, it is important to understand whether the cloning procedure can reverse cellular aging and generate clones with normal genetic and physiological age, similar to those produced from natural reproduction. Telomere shortening is believed to correlate with cellular aging both in vitro and in vivo. Telomere lengths in cells of cloned individuals thus may reflect their genetic age. However, controversies have developed over whether the eroded telomere in somatic cells used for nuclear transfer can be restored during the cloning process. PMID- 16218849 TI - Academic industry relationships in biotechnology: a primer on policy and practice. AB - Academic industry relationships (AIRs) in biotechnology are part of the modern life science economy. They cannot and should not be prevented. But their benefits should not be exaggerated, nor their risks minimized or ignored. It is essential that these relationships be understood, monitored, and managed in a manner that protects the investments and the integrity of involved individuals, institutions, and science in general. Failure to do so could result in loss of public confidence and support for research enterprise--a priceless resource whose integrity and independence are critical to the future of the scientific endeavor. PMID- 16218850 TI - Ethical issues in transgenics. AB - The arguments of critics and concerns of the public on generating transgenic cloned animals are analyzed for the absence or presence of logical structure. Critics' arguments are symbolically compared with "genetic trespassing," "genetic speeding," or "going the wrong way," and responses are provided to these arguments. Scientists will be empowered to participate in the public discussion and to engage the critics on these issues as they consider thoughtful, plausible responses to their concerns. Temporary moratoriums are recognized as a plausible approach to dealing with possible concerns of new scientific advancements. PMID- 16218852 TI - Why are we cloning around . . . ? PMID- 16218853 TI - Bovine blastocysts obtained from reconstructed cytoplast and karyoplasts using a simple portable CO2 incubator. AB - To enable both the multiplication of elite livestock and the engineering of transgenic animals for various agricultural and biochemical purposes, scientists around the world are intensively studying efficient ways of improving developmental competency of bovine embryos reconstructed by somatic cell nuclear transfer. Because it is widely accepted that culture conditions along with many other factors contribute to the developmental competency of reconstructed embryos, this preliminary study was designed to test whether or not bovine reconstructed embryos could develop in vitro using a simple portable CO(2) incubator. CO(2) and O(2) gas tensions and air pressure can be varied using this system. The parameters used in the five conducted trials were low CO(2) (2-5%) and O(2) (8-10%) gas tensions, and negative air pressure (of 300 mm Hg). Chamber temperature was maintained at 38.5 degrees C. Bovine fetal fibroblasts were used as donor karyoplasts and were fused into microsurgically enucleated M II oocytes followed by activation and culture. From the 250 enucleated oocytes, 217 (86.8%) fused, 183 (73.2%) cleaved, and 43 (17.2%) developed to the blastocyst stage. While relatively low developmental rates were achieved, technical proficiency may have been a contributing factor. Further studies using this system are needed to determine optimal levels of O(2), CO(2), and air pressure. PMID- 16218854 TI - Improvement in development of porcine embryos reconstituted with cells from blastocyst-derived cell lines and enucleated oocytes by optimization of reconstruction methods. AB - The present study was conducted to establish the most suitable system for producing porcine reconstructed embryos by transferring cells from blastocyst derived cell lines into enucleated oocytes. When the cells were fused to preactivated metaphase II oocytes, or the cells and arrested metaphase II oocytes were fused in medium without CaCl(2) and MgSO(4), the percentages (43-53%) of fused embryos were significantly lower than those (72-79%) produced by fusing the cells to arrested metaphase II oocytes in medium containing CaCl(2) and MgSO(4). High productive efficiency (7%) of blastocysts was obtained when reconstituted embryos produced by the last method were activated again at 3 hours after fusion (F/A --> Activation). Pronuclear formation was observed in 80-91% of the reconstructed embryos produced by F/A --> Activation, with no significant differences between different culture periods in the medium containing cytochalasin B. When cultured in the medium containing cytochalasin B for 0-1 h, almost all (83-85%) the embryos had one pronucleus and one polar body. However, the number of embryos with two pronuclei and no polar bodies was increased significantly by culturing in the medium containing cytochalasin B for 2-4 h. The cleavage rate (34-48%) of reconstructed embryos was not affected by the presence of cytochalasin B for 2 h after activation. However, the percentage of embryos that developed to the blastocyst stage was significantly higher in the presence (23%) than absence (5%) of cytochalasin B. The results indicate that F/A --> Activation and cytochalasin B treatment are effective for the production of porcine embryos reconstituted with cells from blastocyst-derived cell lines and enucleated oocytes. PMID- 16218855 TI - Activity of maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK) after parthenogenetic activation of ovine oocytes. AB - The maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK) are the key regulators of both meiotic and mitotic cell cycles. Knowledge of the dynamics of these two kinases during the transition from meiosis to mitosis would be of great importance for cloning by nuclear transfer. In this study, experiments were designed to assay the changes of MPF and MAP kinase activity of in vitro matured ovine oocytes after chemical activation and culture in 0 mM or 2 mM 6-dimethylaminopurine (6-DMAP) for 12 h. Moreover, to determine the biological significance of the fluctuations of MPF, activated oocytes were fused with GV staged partners. The biochemical results showed that the high MPF activity of MII oocytes fell to basal level precipitously within the first hour after activation, started to increase at 6-8 h, rising to 80 +/- 4% of MII after 12 h. MAPK activity decreased to a low level 4 h after activation, increased between 6-12 h, but remained below 30 +/- 3.6% of MII values. The incubation with 6-DMAP had no effect on the kinetics of MPF and MAP kinase activity. Fusion of MII oocytes to GV partners induced rapid breakdown of the GV, whereas no breakdown occurred when GV were fused with eggs in the first hours post activation. Interestingly, the high biochemical levels of MPF activity at 8-12 h after activation were not able to induce GVBD in fusion partners. PMID- 16218856 TI - Effects of growth factors and feeder cells on porcine primordial germ cells in vitro. AB - As embryonic stem (ES) cells are not available in swine, embryonic germ (EG) cells derived from primordial germ cells (PGCs) are an alternate source of pluripotent embryonic cells for genetic modification through homologous recombination. Although morphological and biochemical characteristics are similar between ES and EG cells, culture conditions are quite different. To optimize the culture condition for the establishment of porcine EG cells, porcine PGCs were cultured in vitro with various combinations of growth factors (leukemia inhibitory factor [LIF], stem cell factor [SCF], and basic fibroblast growth factor [bFGF]) and on different kinds of feeder cells (STO, TM(4), Sl/Sl(4) m220, porcine embryonic fibroblasts, and COS-7 cells). Optimal results were obtained when all three growth factors (LIF, SCF, and bFGF) were present in the media. Also, feeder cells expressing membrane-bound SCF are required for survival and establishment of porcine EG cells. Therefore, a combination of growth factors and proper feeder cells are critical for the establishment of undifferentiated porcine EG cells. PMID- 16218858 TI - Government encouragement for therapeutic cloning. PMID- 16218859 TI - Developmental competence of nuclear transfer cow oocytes after direct injection of fetal fibroblast nuclei. AB - We have examined the in vitro and in vivo development of cloned embryos produced by incorporation of fetal fibroblast into in vitro matured and enucleated cow oocytes by direct injection and by fusion. For injection, nuclei were either mechanically isolated using the microinjection needle or chemically isolated by treatment with NP-40 lysis buffer. Fetal fibroblasts were serum starved and treated with calcium ionophore before injection to induce chromatin condensation. A range of 8% to 16% of successfully injected oocytes developed to blastocysts in culture and a total of nine pregnancies resulted from transfer of cloned embryos produced by this method. Nuclear transfer by fusion resulted in 22% development to blastocysts. Unlike in mice, the embryos derived from injection did not result in viable pregnancies, which may suggest species differences. All pregnancies were terminated after 45 to 150 days from transfer. Two pregnancies resulted from transfer of cloned embryos obtained by fusion which produced two healthy female calves. The study proposes an alternative method for the production of cow cloned embryos. Further research, however, is required to optimize bovine cloning by injection. PMID- 16218860 TI - Ploidy of bovine nuclear transfer blastocysts reconstructed using in vitro produced blastomere donors. AB - The higher rate of embryonic loss in nuclear transfer compared to in vitro produced embryos may be due to chromosome abnormalities that occur during preimplantation in vitro development. Because little is known about ploidy errors in nuclear transfer embryos, this was examined using embryos reconstructed from in vitro produced embryo donors. In vitro matured oocytes were enucleated and then activated using calcium ionophore A23187 followed by 6-dimethylaminopurine (6-DMAP). Subsequently, embryos were reconstructed using blastomeres from day 4-5 in vitro produced donors. The embryos were cultured until day 7 at which time blastocyst nuclei were extracted and chromosome abnormalities were evaluated by fluorescent in situ hybridization using two probes that bind to the subcentromeric regions on chromosomes 6 and 7. In 16 nuclear transfer blastocysts generated from 5 donor embryos, 53.8 +/- 20.2 (mean % +/- SD) nuclei/embryo were examined. Of these 16, 7 embryos (43.8%) were potentially abnormal because in these, 1.1%, 1.4%, 5.3%, 7.5%, 26.3%, 30.4%, and 66.2% % of the nuclei had a chromosome composition deviating from the diploid condition, indicating a wide degree of variation between embryos. These errors comprised mainly triploid (8.2 +/- 10.3 [0-26.3]: % +/- SD [range]) and tetraploid (10.6 +/- 19.9 [0-54.9]) nuclei with other ploidy combinations accounting for only 0.9 +/- 2.1 [0-2.1]% of deviant nuclei. The proportion of completely normal nuclear transfer embryos was no less than those produced by in vitro fertilization but the distribution of chromosome abnormalities was different (p = 0.0002). In conclusion, nuclear transfer embryos reconstructed using blastomere cells can produce over 50% blastocysts with a diploid chromosome complement. However, the contribution of chromosome abnormalities to embryonic loss in the remaining embryos deserves further investigation. PMID- 16218861 TI - Surface ultrastructural alterations of bovine oocytes after parthenogenetic activation. AB - Oocyte activation is a critical component of the current animal cloning scheme. This study was designed to examine surface characteristics of bovine oocytes by scanning electron microscopy (SEM) after activation by calcium ionophore A23187 (A23187) and electric pulse combined with cycloheximide (CHX) or 6 dimethylaminopurine (6-DMAP) treatments. In vitro matured (IVM) oocytes were activated then harvested at 0 to 19 hours after the onset of treatments for SEM processing. The zona pellucida (ZP) of untreated IVM oocytes exhibited an open mesh structure. The ZP surface showed little changes after A23187 alone, but dramatically changed to a less porous surface 3 hours after combined treatments with CHX or 6-DMAP. The vitelline membrane of IVM oocytes was covered with well developed microvilli (MV). The MV became shorter (0.83 vs. 1.35 microm, p < 0.01) 8 hours after A23187 treatment alone. The vitelline membrane was altered in all oocytes examined 3 hours after incubation with A23187 plus CHX or 6-DMAP. A 1.5 fold increase in the diameter of MV in the CHX group and a higher incidence of large cytoplasmic protrusions (more than 1 microm width) in the 6-DMAP group were observed. After removal of inhibitors, the surface morphologies of the ZP and vitelline membrane were returned nearly to those of untreated IVM oocytes in both groups. The present study clearly showed that surface characteristics of the bovine oocyte were more profoundly changed by a combination of agents for parthenogenetic stimulation, and that the ultrastructural effects were reversible. PMID- 16218862 TI - Cloning of an endangered species (Bos gaurus) using interspecies nuclear transfer. AB - Approximately 100 species become extinct a day. Despite increasing interest in using cloning to rescue endangered species, successful interspecies nuclear transfer has not been previously described, and only a few reports of in vitro embryo formation exist. Here we show that interspecies nuclear transfer can be used to clone an endangered species with normal karyotypic and phenotypic development through implantation and the late stages of fetal growth. Somatic cells from a gaur bull (Bos gaurus), a large wild ox on the verge of extinction, (Species Survival Plan < 100 animals) were electrofused with enucleated oocytes from domestic cows. Twelve percent of the reconstructed oocytes developed to the blastocyst stage, and 18% of these embryos developed to the fetal stage when transferred to surrogate mothers. Three of the fetuses were electively removed at days 46 to 54 of gestation, and two continued gestation longer than 180 (ongoing) and 200 days, respectively. Microsatellite marker and cytogenetic analyses confirmed that the nuclear genome of the cloned animals was gaurus in origin. The gaur nuclei were shown to direct normal fetal development, with differentiation into complex tissue and organs, even though the mitochondrial DNA (mtDNA) within all the tissue types evaluated was derived exclusively from the recipient bovine oocytes. These results suggest that somatic cell cloning methods could be used to restore endangered, or even extinct, species and populations. PMID- 16218863 TI - Stem Cell Research: Stem Cell Research: Medical Progress with Responsibility: Executive Summary. PMID- 16218864 TI - Government response to the recommendations made in the Chief Medical Officer's Expert Group report: "Stem Cell Research: Medical Progress with Responsibility". PMID- 16218867 TI - Radical-mediated dehydration reactions in anaerobic bacteria. AB - Most dehydratases catalyse the elimination of water from beta-hydroxy ketones, beta-hydroxy carboxylic acids or beta-hydroxyacyl-CoA. The electron-withdrawing carbonyl functionalities acidify the alpha-hydrogens to enable their removal by basic amino acid side chains. Anaerobic bacteria, however, ferment amino acids via alpha- or gamma-hydroxyacyl-CoA, dehydrations of which involve the abstraction of a beta-hydrogen, which is ostensibly non-acidic (pK ca. 40). Evidence is accumulating that beta-hydrogens are acidified via transient conversion of the CoA derivatives to enoxy radicals by one-electron transfers, which decrease the pK to 14. The dehydrations of (R)-2-hydroxyacyl-CoA to (E)-2 enoyl-CoA are catalysed by heterodimeric [4Fe-4S]-containing dehydratases, which require reductive activation by an ATP-dependent one-electron transfer mediated by a homodimeric protein with a [4Fe-4S] cluster between the two subunits. The electron is further transferred to the substrate, yielding a ketyl radical anion, which expels the hydroxyl group and forms an enoxy radical. The dehydration of 4 hydroxybutyryl-CoA to crotonyl-CoA involves a similar mechanism, in which the ketyl radical anion is generated by one-electron oxidation. The structure of the FAD- and [4Fe-4S]-containing homotetrameric dehydratase is related to that of acyl-CoA dehydrogenases, suggesting a radical-based mechanism for both flavoproteins. PMID- 16218868 TI - Heterodisulfide reductase from methanogenic archaea: a new catalytic role for an iron-sulfur cluster. AB - Heterodisulfide reductase (HDR) from methanogenic archaea is an iron-sulfur protein that catalyzes reversible reduction of the heterodisulfide (CoM-S-S-CoB) of the methanogenic thiol-coenzymes, coenzyme M (CoM-SH) and coenzyme B (CoB-SH). Via the characterization of a paramagnetic reaction intermediate generated upon oxidation of the enzyme in the presence of coenzyme M, the enzyme was shown to contain a [4Fe-4S] cluster in its active site that catalyzes reduction of the disulfide substrate in two one-electron reduction steps. The formal thiyl radical generated by the initial one-electron reduction of the disulfide is stabilized via reduction and coordination of the resultant thiol to the [4Fe-4S] cluster. PMID- 16218869 TI - Structural and functional comparison of HemN to other radical SAM enzymes. AB - Radical SAM enzymes have only recently been recognized as an ancient family sharing an unusual radical-based reaction mechanism. This late appreciation is due to the extreme oxygen sensitivity of most radical SAM enzymes, making their characterization particularly arduous. Nevertheless, realization that the novel apposition of the established cofactors S-adenosylmethionine and [4Fe-4S] cluster creates an explosive source of catalytic radicals, the appreciation of the sheer size of this previously neglected family, and the rapid succession of three successfully solved crystal structures within a year have ensured that this family has belatedly been noted. In this review, we report the characterization of two enzymes: the established radical SAM enzyme, HemN or oxygen-independent coproporphyrinogen III oxidase from Escherichia coli, and littorine mutase, a presumed radical SAM enzyme, responsible for the conversion of littorine to hyoscyamine in plants. The enzymes are compared to other radical SAM enzymes and in particular the three reported crystal structures from this family, HemN, biotin synthase and MoaA, are discussed. PMID- 16218870 TI - New glycyl radical enzymes catalysing key metabolic steps in anaerobic bacteria. AB - During the last decade, an increasing number of new enzymes containing glycyl radicals in their active sites have been identified and biochemically characterised. These include benzylsuccinate synthase (Bss), 4 hydroxyphenylacetate decarboxylase (Hpd) and the coenzyme B12-independent glycerol dehydratase (Gdh). These are involved in metabolic pathways as different as anaerobic toluene metabolism, fermentative production of p-cresol and glycerol fermentation. Some features of these newly discovered enzymes are described and compared with those of the previously known glycyl radical enzymes pyruvate formate-lyase (Pfl) and anaerobic ribonucleotide reductase (Nrd). Among the new enzymes, Bss and Hpd share the presence of small subunits, the function of which in the catalytic mechanisms is still enigmatic, and both enzymes contain metal centres in addition to the glycyl radical prosthetic group. The activating enzymes of the novel systems also deviate from the standard type, containing at least one additional Fe-S cluster. Finally, the available whole-genome sequences of an increasing number of strictly or facultative anaerobic bacteria revealed the presence of many more hitherto unknown glycyl radical enzyme (GRE) systems. Recent studies suggest that the particular types of these enzymes represent the ends of different evolutionary lines, which emerged early in evolution and diversified to yield remarkably versatile biocatalysts for chemical reactions that are otherwise difficult to perform in anoxic environments. PMID- 16218871 TI - Unusual reactions involved in anaerobic metabolism of phenolic compounds. AB - Aerobic bacteria use molecular oxygen as a common co-substrate for key enzymes of aromatic metabolism. In contrast, in anaerobes all oxygen-dependent reactions are replaced by a set of alternative enzymatic processes. The anaerobic degradation of phenol to a non-aromatic product involves enzymatic processes that are uniquely found in the aromatic metabolism of anaerobic bacteria: (i) ATP dependent phenol carboxylation to 4-hydroxybenzoate via a phenylphosphate intermediate (biological Kolbe-Schmitt carboxylation); (ii) reductive dehydroxylation of 4-hydroxybenzoyl-CoA to benzoyl-CoA; and (iii) ATP-dependent reductive dearomatization of the key intermediate benzoyl-CoA in a 'Birch-like' reduction mechanism. This review summarizes the results of recent mechanistic studies of the enzymes involved in these three key reactions. PMID- 16218872 TI - Novel bacterial molybdenum and tungsten enzymes: three-dimensional structure, spectroscopy, and reaction mechanism. AB - The molybdenum enzymes 4-hydroxybenzoyl-CoA reductase and pyrogallol phloroglucinol transhydroxylase and the tungsten enzyme acetylene hydratase catalyze reductive dehydroxylation reactions, i.e., transhydroxylation between phenolic residues and the addition of water to a triple bond. Such activities are unusual for this class of enzymes, which carry either a mononuclear Mo or W center. Crystallization and subsequent structural analysis by high-resolution X ray crystallography has helped to resolve the reaction centers of these enzymes to a degree that allows us to understand the interaction of the enzyme and the respective substrate(s) in detail, and to develop a concept for the respective reaction mechanism, at least in two cases. PMID- 16218873 TI - Spectroscopic and theoretical approaches for studying radical reactions in class I ribonucleotide reductase. AB - Ribonucleotide reductases (RNRs) catalyze the production of deoxyribonucleotides, which are essential for DNA synthesis and repair in all organisms. The three currently known classes of RNRs are postulated to utilize a similar mechanism for ribonucleotide reduction via a transient thiyl radical, but they differ in the way this radical is generated. Class I RNR, found in all eukaryotic organisms and in some eubacteria and viruses, employs a diferric iron center and a stable tyrosyl radical in a second protein subunit, R2, to drive thiyl radical generation near the substrate binding site in subunit R1. From extensive experimental and theoretical research during the last decades, a general mechanistic model for class I RNR has emerged, showing three major mechanistic steps: generation of the tyrosyl radical by the diiron center in subunit R2, radical transfer to generate the proposed thiyl radical near the substrate bound in subunit R1, and finally catalytic reduction of the bound ribonucleotide. Amino acid- or substrate-derived radicals are involved in all three major reactions. This article summarizes the present mechanistic picture of class I RNR and highlights experimental and theoretical approaches that have contributed to our current understanding of this important class of radical enzymes. PMID- 16218874 TI - Biomimetic metal-radical reactivity: aerial oxidation of alcohols, amines, aminophenols and catechols catalyzed by transition metal complexes. AB - The contributions of the authors to the research program 'Radicals in Enzymatic Catalysis' over the last ca. 5 years are summarized. Significant efforts were directed towards the design and testing of phenol-containing ligands for synthesizing radical-containing transition metal complexes as potential candidates for catalysis of organic substrates like alcohols, amines, aminophenols and catechols. Functional models for different copper oxidases, such as galactose oxidase, amine oxidases, phenoxazinone synthase and catechol oxidase, are reported. The copper complexes synthesized can mimic the function of the metalloenzymes galactose oxidase and amine oxidases by catalyzing the aerial oxidation of alcohols and amines. Even methanol could be oxidized, albeit with a low conversion, by a biradical-copper(II) compound. The presence of a primary kinetic isotope effect, similar to that for galactose oxidase, provides compelling evidence that H-atom abstraction from the alpha-C-atom of the substrates is the rate-limiting step. Although catechol oxidase and phenoxazinone synthase contain copper, manganese(IV) complexes containing radicals have been found to be useful to study synthetic systems and to understand the naturally occurring processes. An 'on-off' mechanism of the radicals without redox participation from the metal centers seems to be operative in the catalysis involving such metal-radical complexes. PMID- 16218875 TI - Combinatorial approaches to functional models for galactose oxidase. AB - The copper enzyme galactose oxidase (GOase, EC 1.1.3.9) catalyses the oxidation of D-galactose and other primary alcohols in air to the corresponding aldehydes and hydrogen peroxide. The current mechanistic hypothesis for this two-electron redox reaction involves a Cu(I)/Cu(II) couple and the reversible oxidation of a ligating phenolate (tyrosine residue of the Tyr272-Cys228 conjugate) to a phenoxyl radical. Our approaches to functional models for galactose oxidase comprise both the use of low-molecular-weight copper complexes of a Schiff-base and sulfonamide ligands, and the synthesis/screening of combinatorial libraries. With regard to the latter, we have synthesized (by the IRORI-directed synthesis approach) peptide libraries carrying either His or the redox-active amino acids Tyr, mod-Cys (a model for the Tyr272-Cys228 conjugate) or TOAC (a TEMPO-derived alpha-amino acid) at four variable positions. After incubation with copper ions, the catalytically active library members were identified by specially designed screening methods. PMID- 16218876 TI - Spectroscopic characterization of the iron-oxo intermediate in cytochrome P450. AB - From analogy to chloroperoxidase from Caldariomyces fumago, it is believed that the electronic structure of the intermediate iron-oxo species in the catalytic cycle of cytochrome P450 corresponds to an iron(IV) porphyrin-pi-cation radical (compound I). However, our recent studies on P450cam revealed that after 8 ms a tyrosine radical and iron(IV) were formed in the reaction of ferric P450 with external oxidants in the shunt pathway. The present study on the heme domain of P450BM3 (P450BMP) shows a similar result. In addition to a tyrosine radical, a contribution from a tryptophan radical was found in the electron paramagnetic resonance (EPR) spectra of P450BMP. Here we present comparative multi-frequency EPR (9.6, 94 and 285 GHz) and Mossbauer spectroscopic studies on freeze-quenched intermediates produced using peroxy acetic acid as oxidant for both P450 cytochromes. After 8 ms in both systems, amino acid radicals occurred instead of the proposed iron(IV) porphyrin-pi-cation radical, which may be transiently formed on a much faster time scale. These findings are discussed with respect to other heme thiolate proteins. Our studies demonstrate that intramolecular electron transfer from aromatic amino acids is a common feature in these enzymes. The electron transfer quenches the presumably transiently formed porphyrin-pi cation radical, which makes it extremely difficult to trap compound I. PMID- 16218877 TI - Impact of Mycoplasma hyorhinis infection on L-arginine metabolism: differential regulation of the human and murine iNOS gene. AB - Infection with mycoplasma is a common problem in cell cultures, with Mycoplasma hyorhinis being the predominant species. Here we investigate the effect of M. hyorhinis infection on L-arginine metabolism, with focus on iNOS-mediated NO synthesis in murine keratinocytes and the human colon cancer cell line DLD-1. iNOS and arginase are L-arginine-metabolizing enzymes involved in the regulation of inflammatory processes, with NO contributing to innate immunity. In murine cells, M. hyorhinis infection enhances cytokine-induced iNOS expression and augments iNOS activity, whereas in the absence of cytokines it causes de novo induction of iNOS mRNA without subsequent translation into iNOS protein. In turn, arginase-1 mRNA expression is diminished in M. hyorhinis-infected murine keratinocytes, resulting in decreased arginase activity. One of the underlying upstream mechanisms is NF-kappaB activation. In contrast, in human cells neither iNOS mRNA nor protein expression is affected by M. hyorhinis infection, but NO synthesis is enhanced, which may be caused by increased L-arginine import. This demonstrates that infection with M. hyorhinis leads to different effects on gene regulation of the murine and human iNOS gene. Our study underlines the importance of routine checking of cell cultures for mycoplasma contamination, particularly in studies on NO-mediated effects or inflammatory processes. PMID- 16218878 TI - Degradation of the sodium taurocholate cotransporting polypeptide (NTCP) by the ubiquitin-proteasome system. AB - The sodium taurocholate cotransporting polypeptide (Ntcp, Slc10a1) is the major uptake system for bile acids into liver cells. This study investigated the degradation of rat Ntcp and human NTCP by the ubiquitin-proteasome system (UPS). In stably transfected HepG2 cells, rat Ntcp was complex-glycosylated and localized at the plasma membrane. Inhibition of proteasomes by MG-132 or lactacystin led to the accumulation of intracellular Ntcp, a process dependent on de novo protein synthesis. Intracellular Ntcp was core-glycosylated, indicating an endoplasmic reticulum (ER) origin. Core-glycosylated Ntcp was found in cytosolic, detergent-insoluble deposits with characteristics of aggresomes: they co-localized with ubiquitin at the microtubule organization center and Ntcp from these deposits was polyubiquitinated. Transient transfections of Ntcp/NTCP induced intracellular deposits that co-localized with ubiquitin, even in the absence of proteasome inhibitors. Similarly, in livers of patients with progressive familial intrahepatic cholestasis, NTCP could be detected co localized with ubiquitin in hepatocytes. We conclude that maturing Ntcp/NTCP is degraded by the ubiquitin-proteasome system at the level of ER-associated degradation (ERAD). An imbalance in the synthesis and degradation of NTCP at the level of the ER or alterations in the ERAD machinery might be the cause of intracellular NTCP deposits in transient transfections and in cholestatic livers. PMID- 16218879 TI - Potential role for immunomodulatory therapy in atherosclerotic plaque stabilisation. AB - Our understanding of the mechanisms underlying acute coronary syndromes has evolved beyond the view that this syndrome reflects a progressive collection of lipids and cellular debris in the vascular wall. Current evidence has implicated a role for inflammation in the pathogenesis of this process. Thus, inflammatory cytokines may attenuate interstitial collagen synthesis, increase matrix degradation and promote apoptosis in several atheroma-associated cell types, and all these cellular events may enhance plaque vulnerability. Recently, a series of experimental studies have reported the plaque-stabilising effects of immunomodulatory therapy such as chemokine blockade, anti-CD40 ligand and IL-10. It is conceivable that some of these approaches will be tested clinically and, if successful, they could provide novel treatment strategies for atherosclerotic plaque stabilisation. PMID- 16218880 TI - Systemic antiatherosclerotic treatment for the peripheral arterial occlusive disease patient. AB - Symptomatic and asymptomatic peripheral arterial occlusive disease (PAD) is indicative of widespread atherosclerosis. The major threat is from cardiovascular ischaemic events; thus, an important therapeutic goal is to modify atherosclerotic risk factors. Data from several large drug trials indicate that patients with PAD may gain as much, if not more, benefit from aggressive secondary prevention than those with other manifestations of atherothrombosis. In fact, the level of care for patients with PAD is lower as compared with those with ischaemic heart disease, although PAD is defined to be a coronary artery risk equivalent. This paper reviews observational data and generalisation from trials in patients with other manifestations of cardiovascular disease that support the importance of treating key risk factors such as smoking, diabetes, hypertension and hyperlipidaemia in PAD patients. PMID- 16218881 TI - Targeting the renin-angiotensin-aldosterone-system in atrial fibrillation: a shift from electrical to structural therapy? AB - Despite its increasing incidence and prevalence, treatment options in atrial fibrillation (AF) are far from ideal and often limited. After decades of focus on the electrical aspects of AF with unsatisfactory results, recent research is focusing increasingly on the atrial structural remodelling that underlies the development of AF in different pathological conditions, such as hypertension, heart failure, diabetes mellitus and coronary artery disease. The aim of this review is to provide a comprehensive overview of the role of the renin angiotensin-aldosterone-system in AF and to highlight the clinical evidence on renin-angiotensin-aldosterone-system blockade as a therapeutic option in AF. PMID- 16218882 TI - Insights into the emerging cardiometabolic prevention and management of diabetes mellitus. AB - Cardiovascular disease (CVD) and Type 2 diabetes mellitus (DM2), once conceived as different entities, share common origins and pathways. Increased activity of the renin-angiotensin-aldosterone-system, insulin resistance, chronic low-grade inflammation and oxidative stress collectively contribute to endothelial dysfunction and atherosclerosis, which manifest clinically as CVD. Nowadays, it is possible to identify and intervene in high-risk populations even before the clinical diagnosis of DM2. The control of dietary patterns and increased physical activity is completely feasible, as well as the management of hypertension and dyslipidaemia. Pharmacological interventions targeted at blocking renin angiotensin-aldosterone-system and sensitising to insulin have a role in the prevention of DM2 and CVD, and are avidly explored worldwide. In the near future, ongoing trials should provide data that will allow us to better treat patients with the cardiometabolic syndrome and diabetes in order to reduce CVD morbidity and mortality. PMID- 16218884 TI - Drug-drug interactions of antifungal agents and implications for patient care. AB - Drug interactions in the gastrointestinal tract, liver and kidneys result from alterations in pH, ionic complexation, and interference with membrane transport proteins and enzymatic processes involved in intestinal absorption, enteric and hepatic metabolism, renal filtration and excretion. Azole antifungals can be involved in drug interactions at all the sites, by one or more of the above mechanisms. Consequently, azoles interact with a vast array of compounds. Drug drug interactions associated with amphotericin B formulations are predictable and result from the renal toxicity and electrolyte disturbances associated with these compounds. The echinocandins are unknown cytochrome P450 substrates and to date are relatively devoid of significant drug-drug interactions. This article reviews drug interactions involving antifungal agents that affect other agents and implications for patient care are highlighted. PMID- 16218883 TI - Cardiovascular system as a 'core' of sexual life. AB - Erectile dysfunction is a common worldwide clinical problem with tens of thousands of new cases per year. It has been argued that erectile dysfunction, like cardiovascular disease and other age-related disorders can be attributed, at least in part, to such modifiable para-aging phenomena. PMID- 16218885 TI - Current options for the treatment of impetigo in children. AB - Impetigo contagiosa is a common, superficial, bacterial infection of the skin characterised by an inflamed and infected epidermis caused by Staphylococcus aureus, Streptococcus pyogenes or both. The less common bullous impetigo is characterised by fragile fluid-filled vesicles and flaccid blisters, and is invariably caused by pathogenic strains of S. aureus. In bullous impetigo, exfoliative toxins are produced, although these are restricted to the area of infection and bacteria can be cultured from the blister contents. In the rare variant, staphylococcal scalded skin syndrome, the exfoliative toxins are spread haematogenously from a localised source causing widespread epidermal damage at distant sites. PMID- 16218886 TI - Antibiotic resistance in Staphylococcus aureus and its relevance in therapy. AB - Staphylococcus aureus is a major cause of infections. Only approximately 20% of the strains remain sensitive to penicillin. Beta-lactamase stable penicillins such as flucloxacillin form the mainstay of treatment of staphylococcal infection. Meticillin-resistant S. aureus (MRSA) are resistant to all beta-lactam antibiotics. Glycopeptide antibiotics are effective against most MRSA strains but, in the last few years, isolates of MRSA that have reduced susceptibility to glycopeptides (glycopeptide-intermediate S. aureus) have been isolated. Some strains exhibit frank resistance to glycopeptides (vancomycin-resistant S. aureus). Infections due to these strains are difficult to treat. This review summarises the therapeutic options for MRSA, glycopeptide-intermediate S. aureus and vancomycin-resistant S. aureus. Novel therapeutic strategies such as immunotherapy and vaccines are also discussed. PMID- 16218887 TI - Syphilis treatment: old and new. AB - Syphilis has challenged scientists and clinicians since its first appearance in the late 1400s and debate continues to surround the best practice in management. Difficulties in defining the goals of successful treatment have contributed to problems in determining recommendations for the ideal management. Treatment regimens currently in use were developed before randomised controlled trials became standard. This, combined with national differences in disease definition, staging and varying interpretations of the studies, as well as the emergence of complicating comorbid conditions, such as HIV, has resulted in a lack of consensus for treatment. This paper will discuss the history and current treatment of syphilis focusing on dilemmas faced by clinicians today, including the emergence of a resistant strain. Despite the difference between current national guidelines, penicillin G largely remains the treatment of choice. Close follow up, monitoring and ensuring adequate compliance remain the most important aspects in the treatment of syphilis. PMID- 16218888 TI - Therapeutic approaches to Chlamydia infections. AB - Chlamydia trachomatis ocular and urogenital infections represent major public health problems, whereas Chlamydophila pneumoniae is a common aetiological agent of community-acquired pneumonia. The obligate intracellular lifestyle of these established pathogens poses challenges to both their diagnosis and treatment. Tetracyclines, macrolides and quinolones remain the antimicrobials of choice for the treatment of infections due to Chlamydiaceae. PMID- 16218889 TI - What can be learned from the experience with the fixed low-dose combination of perindopril/indapamide in the treatment of hypertension? AB - The association of low doses of perindopril and indapamide in a single pill has been developed to meet the criteria required for a fixed-dose combination to be used as first-line therapy. The experience accumulated so far has demonstrated a greater antihypertensive efficacy of this preparation compared with various monotherapies, but not at the expense of a worsening of tolerability. The perindopril/indapamide combination is particularly effective in lowering systolic and pulse blood pressure. Starting treatment with this fixed-dose combination improves arterial stiffness and allows a better regression of cardiac hypertrophy than angiotensin-converting enzyme-inhibition or beta-adrenoceptor blockade alone. In hypertensive patients with Type 2 diabetes, a greater reduction of urinary albumin excretion can be obtained with the perindopril/indapamide association compared with an angiotensin-converting enzyme inhibitor administered as monotherapy. There is evidence that a first-line management of hypertension based on a low-dose perindopril/indapamide combination can provide a better efficacy/safety ratio than other well-established therapeutic strategies. PMID- 16218890 TI - Oral ibandronate: a less frequently administered therapeutic option for postmenopausal osteoporosis. AB - Osteoporosis is a severe condition, associated with significant disability as a result of fragility fractures and increased mortality. Oral bisphosphonates effectively reduce the risk of osteoporotic fracture and are generally well tolerated. Unfortunately, patient outcomes are often compromised by suboptimal therapeutic adherence. In other disease areas, reduced dosing frequency has been shown to improve therapeutic adherence. A positive impact for adherence has been observed with a reduction in the bisphosphonate dosing frequency from daily to weekly. However, overall adherence remains suboptimal. Ibandronate is a potent nitrogen-containing bisphosphonate specifically designed for less frequent than weekly administration, without compromise for efficacy or tolerability. This article reviews the pharmacology, efficacy and tolerability of oral ibandronate when administered with extended dosing intervals in postmenopausal osteoporosis. PMID- 16218892 TI - Alefacept: an expert review concerning the treatment of psoriasis. AB - Psoriasis is a chronic, inflammatory skin disease. Historically, phototherapy or immunosuppressive agents have been the first line of treatment for patients with severe psoriasis; however, the long-term use of these agents is limited by dose dependent toxicities. Alefacept was the first biological agent approved in both the US and Canada for the treatment of adults with moderate-to-severe chronic plaque psoriasis. Alefacept is a remittive therapy that selectively reduces memory T cells. The efficacy and safety of up to two courses of alefacept have been demonstrated in clinical trials, and thus, this review focuses on new data to optimise the use of this biological agent. Emerging data indicates that multiple courses of alefacept for the long-term treatment of psoriasis are safe and effective. In addition, data are reviewed on the use of alefacept in combination with other agents and in other diseases, including psoriatic arthritis. PMID- 16218891 TI - Update on linezolid: the first oxazolidinone antibiotic. AB - Linezolid is the first of an entirely new class of antibiotics, the oxazolidinones, in decades. It has a spectrum of activity against virtually all important Gram-positive pathogens. The unique mechanism of action of linezolid makes cross-resistance with other antimicrobial agents unlikely. Linezolid has both intravenous and oral formulations and the latter is 100% bioavailable. Since its first approval and marketing in March 2000 in the US, linezolid has gained approval for use in many other countries for the treatment of community-acquired and nosocomial pneumonia, complicated and uncomplicated skin and soft-tissue infections, and infections caused by methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci, including cases with concurrent bacteraemia. Several earlier comprehensive reviews summarised the chemistry, mechanism of action, pharmacokinetics, clinical efficacy and safety profile of linezolid. The present review provides an update on the latest data regarding the antimicrobial activity of linezolid versus other commonly used agents, the clinical and health-economic outcomes of linezolid versus vancomycin and teicoplanin, and safety issues. PMID- 16218893 TI - Azithromycin and lower respiratory tract infections. AB - Azithromycin is a macrolide antibiotic that has been structurally modified from erythromycin with an expanded spectrum of activity and improved tissue pharmacokinetic characteristics relative to erythromycin. This allows once-daily administration for 3-5 days of treatment compared with traditional multi dosing 7 10-day treatment regimens. It has been successfully employed in lower respiratory tract infections. Recent data indicate that azithromycin may exert anti inflammatory/immunomodulatory effects that may be of use in the treatment of both acute and chronic airway diseases. This review examines the role of azithromycin in lower respiratory tract infections analysing published data on exacerbations of chronic bronchitis, community-acquired pneumonia and cystic fibrosis both in adults and children. In addition, pharmacokinetic and pharmacodynamic properties of the drug are also considered. PMID- 16218895 TI - More support for high-dose atorvastatin in coronary artery disease. PMID- 16218894 TI - A review of exemestane in the management of breast cancer. AB - Breast cancer is a major health problem in developed countries. Endocrine therapy is a key component in the management of hormone receptor-positive disease. Although tamoxifen has historically been the gold standard in the first-line management of early and advanced breast cancer, the rise of third-generation aromatase inhibitors in the past decade has resulted in a major shift in endocrine therapy. Clinical trials of aromatase inhibitors including exemestane, an orally active steroidal aromatase inactivator, have demonstrated significant improvements in outcome measures compared with tamoxifen. In early breast cancer, key questions remain regarding the optimal sequence, duration and type of aromatase inhibitors, as well as their long-term safety. PMID- 16218896 TI - A Tribute to William B. Long, Jr., and William B. Long, III: A Celebration of Their Revolutionary Contributions to Trauma Care. AB - An emergency medical system for trauma care has been conceived in our nation in an effort to improve delivery of emergency care to the accidentally injured patient. There are an estimated 20 million disabling injuries in our nation that should be cared for in trauma centers each year. This report has been written to acknowledge Dr. William Long, Jr., as well as Dr. William B. Long, III, for their unique contributions in establishing the Maryland Statewide Trauma System. Dr. William Long, Jr., played an instrumental role in working with Dr. R Adams Cowley to verify the life-saving value of the Maryland State Police helicopter system. In addition, Dr. Long, Jr., crafted a plan with Dr. R Adams Cowley that allowed Dr. Cowley the autonomy from the University of Maryland Medical School to develop a separate and distinct trauma facility, which is recognized throughout the world. It is indeed fortuitous that Dr. William B Long, III, experienced these landmark changes in trauma care in Maryland, which provided a catalyst for his future career that included extensive training in general surgery in Edinburgh as well as training in trauma surgery with Dr R Adams Cowley. These unique experiences convinced him to expand his training into cardiothoracic surgery. During these academic adventures, he became an international authority on the mathematics of trauma scores, cardiothoracic trauma resuscitation, and the components of a Level I trauma center. These empowering experiences became a catalyst for Dr. William Long, III, to undertake the scientific and clinical studies that would allow him to develop the only American College of Surgeons Committee on Trauma (ACSCOT) Verified Level I Trauma Center in the Pacific Northwest. This report describes in considerable detail Dr. William B. Long, III,'s Trauma Center at Legacy Emanuel Trauma Center (Portland, Oregon) as well as to outline his plans to further improve trauma care in the state of Oregon so that it remains a legacy for his academic career. His dreams for having a comprehensive trauma system in the Pacific Northwest are described in detail so that it an be replicated in our nation and our world.Dr. Long became the Trauma Medical Director for Emanuel Hospital in the Fall of 1983. He began building Emanuel's trauma program by establishing an infrastructure that would support technically advanced ways of restoring life and function. His trauma center consisted of the following components: trauma registry, trauma resuscitation nurse program, direct to operating room policy with unstable trauma patients, anesthesia as part of the trauma resuscitation team, massive transfusion protocol, mobile surgical transport team, outreach to rural communities, recruitment of specialists with interest in trauma care, development of a new trauma physical facility, and the Physician Assistant educational program. PMID- 16218897 TI - Device-related infections: a review. AB - The use of surgically implanted devices has increased as a result of their beneficial effect on quality of life, and in some circumstances, on patient survival rates. They can, however, be associated with a variety of complications, the most dreaded being infection. Device-related infections are important to understand because of the morbidity and mortality associated with them. Frequently, patients are managed with hospitalization, prolonged courses of antibiotics, and surgical interventions, all of which can negatively impact on patients' quality of life. Such care is also associated with increased costs to health care systems. Furthermore, these infections often represent a diagnostic challenge because of the lack of consensus definition of what constitutes an infection and its severity, as well as the paucity of well-designed, large studies addressing optimal methods of investigation and management. An implant associated infection is defined as a host immune response to one or more microbial pathogens on an indwelling implant. An understanding of the pathogenesis of these infections provides a rationale for management. Development of device-related infections begins with colonization of the foreign material, followed by a complex metamorphosis by the microorganisms with resultant biofilm formation. In this surface-associated form, bacteria have altered phenotypic properties. This change, in conjunction with the physical protective layer provided by the biofilm, renders antimicrobial therapy ineffective when used alone. Because the microorganisms are able to reside on the hardware, they proliferate and cause local damage, such as loosening of implanted devices, wound dehiscence, or disruption of prosthetic valves, as well as systemic manifestations, such as fever or embolic phenomenon. The onset and clinical manifestations of device-related infections vary with the pathogen involved, as well as which component of the device is affected. The time period after device implantation that signs and symptoms develop can assist in the selection of empiric antimicrobial therapy. Optimal diagnostic microbiologic specimens are paramount in tailoring the antimicrobial therapy, which almost always has to be given for a prolonged period of time. Surgical removal of the device is usually necessary. Some studies of limited types of device-related infections, however, have defined indications for which salvage therapy may be warranted. In addition, some patients are not candidates for, or may not want, further surgical interventions, in which case indefinite suppressive antimicrobial therapy may be considered. This review provides an overview of infections related to various neurosurgical, cardiac, and orthopedic devices, as well as those related to cochlear, breast, and penile prostheses, with discussion of definitions of such infections, along with microbiology, pathogenesis, and management guidelines, including the limited indications for salvage techniques. PMID- 16218898 TI - Histologic and histomorphometric analysis of an immediately loaded implant retrieved from man after 14 months of loading. AB - OBJECTIVE: The aim of this study was a histologic and histomorphometric analysis of the peri-implant tissue reactions and the bone/titanium interface in an immediately loaded titanium implant inserted in a soft bone site and retrieved, in man, after a 14-month loading period. METHODS. A 65-year-old patient presented with a partial edentulism in the left posterior mandible. The patient was rehabilitated with three dental implants inserted in the left first, second, and third molar sites. All these implants were immediately put into a nonfunctional loading mode and joined with the other implants that supported the temporary restorations. After 14 months, the most distal implant and surrounding tissues were retrieved because of psychological problems of the patient. RESULTS: Newly formed, strongly stained, compact, mature cortical bone with few marrow spaces was observed around the implant, especially in the coronal portion. No inflammatory infiltrate was present around the implant. No gaps or dense fibrous connective tissue were found at the bone/metal interface. No apical epithelial migration was found. In the cortical portion, bone remodeling areas were present with many newly formed Haversian canals. Only in a few areas of the interface was it possible to observe an osteoblast rim. In the apical portion, newly formed bone trabeculae were present; these were composed mostly by woven bone, and only a small quantity of preexisting lamellar bone was present. Histomorphometric evaluation showed that the bone/implant contact percentage was 72.6% (+/-2.7%). CONCLUSIONS: We found that, in immediately loaded implants inserted in soft tissue sites, it was possible to find a high percentage of bone-to-implant contact and that osseointegration was maintained for more than 1 year. This fact could be partly explained with the use of an implant with a rough surface. PMID- 16218899 TI - Bacterial diseases of the skin. AB - When considering common bacterial diseases of the skin, rather distinct clinical responses to a variety of bacterial infections have been identified. In these cases, it is the specific site of infection and the attendant inflammatory responses that provide the characteristic clinical picture. When the pyoderma extends just below the stratum corneum, it is called impetigo. Nonbullous impetigo is the most common pediatric skin infection. It usually starts in a traumatized area. The typical lesion begins as an erythematous papule, after which it becomes a unilocular vesicle. When the subcorneal vesicle becomes pustular, it ruptures and eventually becomes a yellow, golden crust that is a hallmark of the disease process. Bullous impetigo is a less common form of impetigo, accounting for fewer than 30% of all impetigo cases. It occurs in infants and is characterized by rapid progression of vesicles to the formation of bullae measuring larger than 5 mm in diameter in previously untraumatized skin. Treatment of nonbullous impetigo must include intervention against the pathogen as well as improvements in the hygiene and living conditions of the patient. A fundamental tenet is to debride the crust (scab) from the wound surface using poloxamer 188. If the lesions are not widespread, topical mupirocin is the treatment of choice. Treatment of bullous impetigo is similar, except that the local cleansing and topical antibiotic must be complemented by systemic antibiotics if there is evidence of disseminating infections. Ecthyma is usually a consequence of failure to treat effectively impetigo. The untreated infection extends deep into the tissue in shallow ulcerations that often heal without scar. Treatment for ecthyma usually requires systemic antibiotics against either staphylococcus or streptococcus. Folliculitis is a pyoderma located within a hair follicle, secondary to follicular occlusion by keratin, overhydration, or either bacterial or fungal infection. Folliculitis may be divided into either a deep or a superficial type. In the superficial type, the pustule is located at the opening of the hair follicle. In the deep form, the infection may extend beyond the confines of the hair follicle, becoming a furuncle or boil. Carbuncles are aggregates of interconnected furuncles that drain through multiple openings of the skin. Treatment of folliculitis must include searching for and avoiding any factors predisposing to infection. If topical antibiotic therapy is ineffective in controlling the infection, surgical drainage of the infected skin abscess will be necessary. Paronychia is the most common bacterial infection of the hand, which often requires surgical incisional drainage. Similarly, a felon that is an infection of the distal pulp of a finger usually requires surgical drainage. Finally, cellulitis is an acute inflammatory reaction involving the skin and underlying subcutaneous tissue. It usually starts as erysipelas and may advance to lymphangitis, lymphadenitis, or gangrene,which will respond to life-saving interventions in the hospital that usually include systemic antibiotic treatment as well as surgical intervention. PMID- 16218900 TI - Modern concepts of treatment and prevention of electrical burns. AB - Electric injuries account for 1,000 deaths in the United States, with a mortality rate of 3--15%. As the widespread use of electricity and injuries from it increase, all health professionals involved in burn care must appreciate its physiological and pathological effects as well as management of electrical current injury. Electric current exists in two forms: alternating current and direct current. The effects of electricity on the body are determined by seven factors: (1) type of current, (2) amount of current, (3) pathway of current, (4) duration of current, (5) area of contact, (6) resistance of the body, and (7) voltage. Electrical accidents can be divided into less than 1,000 V (low-voltage accidents) and greater than 1,000 V (high-voltage accidents). In any electrical accident, the witness must turn off the power source and initiate treatment at the scene of the injury. Low-voltage electric burns almost exclusively involve either the hands or oral cavity. Surgical treatment will vary with the severity of the injury. Burns caused by contact with a high-voltage alternating electric circuit conforms to two types: burns from an electric arc and burns from an electric current. High-voltage electric current injuries have a wide variety of systemic manifestations, including neurologic complications, cardiovascular and pulmonary manifestations, vascular damage, and abdominal, bone, eye and joint complications. An organized approach to the management of these complications is outlined in this article. The best treatment of burn injuries remains prevention. Because the majority of burn injuries are due to occupational electrical injuries, the regional burn centers must work effectively with industry to prevent these potentially life-threatening accidents. PMID- 16218901 TI - Breast cancer and ovarian cancer genetics. AB - Breast and ovarian cancers are the second and fifth leading causes of cancer death, respectively, among women in the United States. Individuals with breast cancer have a 20--30% chance of having at least one relative with the disease. However, only 5--10% of the cases are a direct result of germline mutations in highly penetrable genes, such as BRCA1 and BRCA2 (BRCA1/2) as well as genes TP53 and PTEN. Since 1996, genetic testing for these mutations has been clinically available. A strategy for the management of women at increased familial risk of breast and ovarian cancers is described, which includes genetic assessment, chemoprevention, radiologic screening, and clinical and self-examination. Genetic testing should occur within a cancer genetic clinic after genetic counseling. A blood sample allows determination of the presence of the BRCA1 and BRCA2 genes, the TP53 gene, the PTEN gene, and the ATM gene. Tumor examination has identified a growth factor receptor gene, human epidermal growth factor receptor (HER-2). With regard to diet and lifestyle, women at increased risk of breast cancer could be advised to reduce dietary fat, avoid obesity, decrease alcohol consumption, and take regular exercise. Although chemoprotection is a valuable consideration, it is important to emphasize that the use of Tamoxifen in BRCA1 and BRCA2 mutation carriers is not established, nor is the optimum duration of benefit. An overview of the main outcomes of the current published studies confirms a 38% decrease in breast cancer incidence with Tamoxifen but recommends its use be restricted to women at high risk of breast cancer and low risk for potential side effects. The role of bilateral risk-reducing mastectomy or prophylactic mastectomy has been controversial for several reasons, including the psychosocial significance of the breast in Western cultures, the wide acceptance of breast conservation in surgery for early breast cancer, and the previous lack of data on its efficacy. The surgical procedure should aim to remove substantially all at risk breast tissue. However, there is a balance between reduction of cancer risk and cosmetic outcome. Bilateral prophylactic oophorectomy can significantly decrease ovarian cancer risk in women who carry BCRA1 mutations. Oophorectomy lowers the risk of breast cancer, even in women who have previously used hormone replacement therapy. There are no published randomized controlled trials examining the effectiveness of mammographic screening in women under 50 years of age with a family history of breast cancer. However, the published studies do suggest that mammographic screening of a high-risk group of women under 50 years of age may detect cancer at a rate equivalent to that seen in women 10 years older with normal risk. Other initial studies also support MRI as having a greater sensitivity than mammography in high-risk women. Breast clinical and self examination is often advocated, but its effectiveness is unproved, and only one randomized study has been undertaken in women at risk. On the basis of this study as well as one nonrandomized study, it can be concluded that clinical examination as well as mammography are essential in detecting breast cancer. under 50 years of age with a family history of breast cancer. However, the published studies do suggest that mammographic screening of a high-risk group of women under 50 years of age may detect cancer at a rate equivalent to that seen in women 10 years older with normal risk. Other initial studies also support MRI as having a greater sensitivity than mammography in high-risk women. Breast clinical and self examination is often advocated, but its effectiveness is unproved, and only one randomized study has been undertaken in women at risk. On the basis of this study as well as one nonrandomized study, it can be concluded that clinical examination as well as mammography are essential in detecting breast cancer. PMID- 16218902 TI - Human acellular dermal matrix for repair of abdominal wall defects: review of clinical experience and experimental data. AB - The use of prosthetic mesh for the tension-free repair of incisional hernias has been shown to be more effective than primary suture repair. Unfortunately, prosthetic materials can be a suboptimal choice in a variety of clinical scenarios. In general, prosthetic materials should not be implanted into sites with known contamination or infection because they lack an endogenous vascular network and are thus incapable of clearing bacteria. This is of particular relevance to the repair of recurrent hernias, which are often refractory to repair because of indolent bacterial colonization that weakens the site and retards appropriate healing. Although fascia lata grafts and muscle flaps can be employed for tension-free hernia repairs, they carry the potential for significant donor site morbidity. Recently, a growing number of clinicians have used human acellular dermal matrix as a graft material for the tension-free repair of ventral hernias. This material has been shown to become revascularized in both animal and human subjects. Once repopulated with a vascular network, this graft material is theoretically capable of clearing bacteria, a property not found in prosthetic graft materials. Unlike autologous materials such as fascial grafts and muscle flaps, acellular dermal matrix can be used without subjecting the patient to additional morbidity in the form of donor site complications. This article presents a thorough review of the current literature, describing the properties of human acellular dermal matrix and discussing both animal and human studies of its clinical performance. In addition to the review of previously published clinical experiences, we discuss our own preliminary results with the use of acellular dermal matrix for ventral hernia repair in 46 patients. PMID- 16218903 TI - Texas passes first law for safe patient handling in America: landmark legislation protects health-care workers and patients from injury related to manual patient lifting. AB - On June 17,2005, Texas Governor Rick Perry (R) signed into law Senate Bill 1525, making Texas the first state in the nation to require hospitals and nursing homes to implement safe patient handling and movement programs. Governor Perry is to be commended for this heroic first stand for safe patient handling in America. The landmark legislation will take effect January 1, 2006, requiring the establishment of policy to identify, assess, and develop methods of controlling the risk of injury to patients and nurses associated with lifting, transferring, repositioning, and movement of patients; evaluation of alternative methods from manual lifting to reduce the risk of injury from patient lifting, including equipment and patient care environment; restricting, to the extent feasible with existing equipment, manual handling of all or most of a patient's weight to emergency, life-threatening, or exceptional circumstances; and provision for refusal to perform patient handling tasks believed to involve unacceptable risks of injury to a patient or nurse. Manually lifting patients has been called deplorable, inefficient, dangerous to nurses, and painful and brutal to patients; manual lifting can cause needless suffering and injury to patients, with dangers including pain, bruising, skin tears, abrasions, tube dislodgement, dislocations, fractures, and being dropped by nursing staff during attempts to manually lift. Use of safe, secure, mechanical lift equipment and gentle friction-reducing devices for patient maneuvering tasks could eliminate such needless brutality. Research has proven that manual patient lifting is extremely hazardous to health care workers, creating substantial risk of low-back injury, whether with one or two patient handlers. Studies on the use of mechanical patient lift equipment, by either nursing staff or lift teams, have proven repeatedly that most nursing staff back injury is preventable, leading to substantial savings to employers on medical and compensation costs. Because the health-care industry has relied on people to do the work of machines, nursing work remains the most dangerous occupation for disabling back injury. Back injury from patient lifting may be the single largest contributor to the nursing shortage, with perhaps 12% of nurses leaving or being terminated because of back injury. The US health-care industry has not kept pace with other industries, which provide mechanical lift equipment for lifting loads equivalent to the weight of patients, or with other countries, such as Australia and England, which are more advanced in their use of modern technology for patient lifting and with no-lifting practices in compliance with government regulations and nursing policies banning manual lifting. With Texas being the first state to succeed in passing legislation for safe patient handling, other states are working toward legislative protection against injury with manual patient lifting. California re-introduced safe patient handling legislation on February 17, 2005, with CA SB 363, Hospitals: Lift Teams, following the September 22, 2004, veto of CA AB 2532 by Governor Arnold Schwarzenegger, who said he believes existing statutory protection and workplace safety standards are sufficient to protect health care workers from injury. Massachusetts HB 2662, Relating to Safe Patient Handling in Certain Health Facilities, was introduced December 1, 2004. Ohio HB 67, signed March 21, 2005 by Governor Bob Taft (R), creates a program for interest-free loans to nursing homes for implementation of a no-manual-lift program. New York companion bills AB 7641 and SB 4029 were introduced in April, 2005, calling for creation of a 2-year study to establish safe patient handling programs and collect data on nursing staff and patient injury with manual patient handling versus lift equipment, to determine best practices for improving health and safety of health-care workers and patients during patient handling. Washington State is planning re introduction of safe patient handling legislation, after WA HB 1672, Relating to reducing injuries among patients and health care workers, was stalled in committee in February, 2005. Language from these state initiatives may be used as models to assist other states with drafting safe patient handling legislation. Rapid enactment of a federal mandate for Safe Patient Handling No Manual Lift is essential and anticipated. PMID- 16218904 TI - The need to legislate the health-care industry in the state of Washington to protect health-care workers from back injury. AB - There is an epidemic of health-care worker back injury in the State of Washington. Voluntary programs are not keeping pace with the increasing back injury rates to health-care workers. Adding all the health-care industry SIC codes, hospitals, nursing homes, home health and residential care puts health care as the leading industry in the State of Washington for back injury. Licensed practical nurses, nurses aides, and registered nurses account for the majority of all claims in the health-care industry. Self-insured hospitals led the state for lost time compensable back injuries between 1993 and 2001, and combined Washington State Fund and self-insured health-care hospitals and nursing homes led all other industries for compensable soft-tissue disorders of the neck, back, and upper extremities. Legislation is needed to protect this group of workers in this type of industry. A bill will be re-introduced in the 2006 session that calls for hospitals to implement back injury prevention programs through either the Zero-Lift model, with nursing staff use of lift equipment, or the Lift-Team model, with a specially trained team using lift equipment, or a combination of the two, for all shifts. The State of Washington should provide funding, through savings created by back injury prevention programs, for small rural health-care institutions to assist them in compliance with the legislation. PMID- 16218905 TI - The need for an organized approach for Government Medical Insurance Programs in the Commonwealth of Virginia. AB - The Commonwealth of Virginia has a disorganized approach to enrolling their retired faculty in Medicare Supplement Insurance Programs. An organized approach to establishing Medicare Supplemental Insurance for retired University faculty should include the following administrative changes to correct this potential health-care crisis for retired state faculty members. First, the ombudsman for human resources for the state universities must receive educational programs that prepare the retired faculty members over the age of 65 to select the corporate insurance policy from Anthem Blue Cross/Blue Shield Insurance Company. Included in this educational program should be a review of the Advantage 65 Member Handbook. Second, they must point out to the faculty member that they are receiving a CORPORATE insurance policy rather than an individual insurance policy from Anthem Blue Cross/Blue Shield Insurance Company. They must provide the telephone numbers of the Anthem Blue Cross/Blue Shield offices in Roanoke, Virginia. Concomitantly, they must send the name and address of the faculty member to the Commonwealth of Virginia Department of Human Resource Management. They should inform the faculty member that the Commonwealth of Virginia Department of Human Resource Management will be sending them newsletters that outline any changes in the corporate insurance policy that they coordinate with the Anthem Blue Cross/Blue Shield Insurance Company. The Commonwealth of Virginia Department of Human Resource Management must take on some new responsibilities in their efforts to coordinate health-care coverage of the retired faculty over the age of 65. First, they must have a computer registry of all corporate health-care policies of the individual faculty members to ensure that newsletters are being sent to them. Ideally, this agency should have a computerized system that allows it to send out its newsletter update by email to those retired faculty members who have computers. They should urge the faculty members to initiate an automatic check payment withdrawal from their bank so that the premiums from their corporate insurance policy can be paid promptly to Anthem Blue Cross/Blue Shield Insurance Company. Whereas the universities and the Commonwealth of Virginia Department of Human Resource Management are making these responsible changes, Anthem Blue Cross/Blue Shield Insurance Company must undertake innovative changes in their corporate health-care policy to assist the retired faculty member. For instance, they must list on the insurance card that the faculty member has a corporate policy. Like the United Health Insurance Company, it would be advisable to offer a 2-5% reduction in the cost of the corporate insurance policy if the faculty member begins an automatic premium payment agreement through their bank. This insurance discount would be an added incentive for the faculty member to do automatic payments through their bank. PMID- 16218906 TI - Micropatterned avidin arrays on silicon substrates via photolithography, self assembly and bioconjugation. AB - Processes for micropatterning protein arrays on inorganic substrates have gained attention in the development of biosensors and clinical diagnostics. This study demonstrates a chemically selective method based on photolithographic deposition of gold patterns with the subsequent attachment of functionalized alkanethiols via molecular self-assembly. Selective capping of carboxy groups on alkanethiols by N-hydroxysuccinimide esters allowed the deposition of methoxypoly(ethylene glycol) silane as a blocking agent on unpatterned regions. Carboxylates were used to couple a form of avidin to create a microarray of protein. This microarray was successfully probed with biotinylated quantum dots. In-process characterization methods included grazing-angle Fourier-transform IR spectroscopy, ellipsometry, contact-angle goniometry, atomic-force microscopy and fluorescence microscopy. PMID- 16218907 TI - Fusion protein between protein ZZ and red fluorescent protein DsRed and its application to immunoassays. AB - In the present study, a red fluorescent protein (DsRed) from the coral Discosoma was fused to the C-terminus of protein ZZ, a synthetic artificial IgG-Fc-fragment binding protein derived from the B-domain of staphylococcal Protein A. The chimaeric protein, tagged with six histidine residues at the N-terminus, was expressed in Escherichia coli and easily purified by one-step Ni2+-chelating affinity chromatography. Its fluorescence and IgG-binding activities were validated using fluorescence-spectrum analysis, ELISA and dot-blot analysis. Furthermore, in subsequent dot-blotting immunoanalysis of glutathione S transferase and tumour necrosis factor-alpha, and immunofluorescent microscopy assay of interferon regulatory factor 3, the chimaeric protein enabled effective detection of target molecules. Compared with fluorescence-conjugated antibodies, ZZ-DsRed is less susceptible to photobleaching and easy to produce. In addition, unlike HRP (horseradish peroxidase)-conjugated antibodies, using ZZ-DsRed needs no addition of a chromogenic reagent. Our results indicate that ZZ-DsRed shows a wide and promising application potential in immunological detection as a substitute for fluorescent or HRP-conjugated anti-IgGs. PMID- 16218908 TI - Epilepsy, osteoporosis and fracture risk - a meta-analysis. AB - This meta-analysis assesses the effects of epilepsy on fracture risk and changes in bone mineral density (BMD) in patients with epilepsy. A search of PubMed was conducted using the key words epilepsy, fracture, and bone mineral. A weighted estimate of relative risk of fractures and changes in BMD (Z-score) was calculated. From the changes in BMD, expected increase in relative fracture risk was calculated. A total of 11 studies on fracture risk and 12 studies on BMD were retrieved. The relative risk of any fracture was increased (2.2, 95% CI: 1.9-2.5, five studies), as was the risk of hip (5.3, 3.2-8.8, six studies), forearm (1.7, 1.2-2.3, six studies), and spine fractures (6.2, 2.5-15.5, three studies). A large proportion of fractures (35%) seemed related to seizures. Spine (mean +/- SEM: -0.38 +/- 0.06) and hip (-0.56 +/- 0.06) BMD Z-scores were significantly decreased, hip more than spine (2P < 0.05). The expected increases in relative risk of any fracture from BMD Z-scores were 1.2-1.3, and significantly lower than observed (2P < 0.05). The deficit in BMD in patients with epilepsy is too small to explain the observed increase in fracture risk. The remainder of the increase in fracture risk may be linked to seizures. PMID- 16218909 TI - PCR in lyme neuroborreliosis: a prospective study. AB - OBJECTIVES: DNA proof is the only widely available direct diagnostic tool in Lyme borreliosis. Sensitive PCR detecting of spirochetal DNA was prepared and a prospective study in neuroborreliosis was performed. MATERIALS AND METHODS: 57 hospitalised patients with active neuroborreliosis and proved CSF antibodies synthesis were examined. Nested-PCR (utilizing three targets) was used for the detection of specific DNA in plasma, CSF and urine. RESULTS: Before treatment 36 positive patients (63.1%) were found in all tested specimens in parallel, 28 patients (49.1%) were positive in urine, 20 in CSF (35.0%) and 16 in plasma 28.0%). Later only urine was tested and the following results were obtained: 17 positive patients (30.0%) immediately after treatment, 8 (14.0%) after 3 months and one patient persisted positivity after 6 months. CONCLUSIONS: The highest sensitivity of PCR was achieved in the acute period of neuroborreliosis - 63.1% in three body fluids comparing with CSF antibody synthesis. PMID- 16218910 TI - Effect of temporary clipping on frontal lobe functions in patients with ruptured aneurysm of the anterior communicating artery. AB - BACKGROUND: After surgery for ruptured anterior communicating artery (ACoA) aneurysm, several patients who have achieved a favorable neurological outcome yet have been observed to suffer from a poor cognitive outcome. The aim of this study was to explore the possible effects of temporary clip applications on frontal lobe functions in the patients with ruptured ACoA aneurysm. METHODS: Forty patients were chosen among a series of cases who underwent an early surgery (within 96 h) after ACoA aneurysm bleeding. All of them were in Hunt-Hess grade 1 or grade 2. Of the 40 patients, temporary clipping was used in 22 patients (group A), whereas it was not used in 18 patients (group B). These two groups were compared with 20 volunteers (group C) without neurologic or psychiatric disorders. RESULTS: The mean duration of temporary vessel occlusion for both A1 was 8.2 +/- 2.9 min (4-15) in group A. Neither clinical nor radiographic strokes were detected. An improvement in frontal lobe function occurred at long term in group B patients. Whereas, cognitive deficits were persisting at long-term follow up in group A, especially in patients who had temporary clipping duration longer than 9 min. CONCLUSIONS: The results emphasize that the negative effects of temporary vessel occlusion on cognitive changes occur before ischemic damage. Thus, such negative effects of temporary clipping on cognitive functions should not be neglected by surgeons during surgery. PMID- 16218911 TI - Idiopathic intracranial hypertension in Dubai: nature and prognosis. AB - OBJECTIVE: To evaluate the nature and course of idiopathic intracranial hypertension (IIH) in Dubai, UAE. METHODS: In a retrospective study the medical records of 50 patients with IIH were reviewed. All of them were diagnosed according to the Modified Dandy Criteria. RESULTS: There were 46 (92%) women. Mean age at presentation was 35.7 years. Obesity was the commonest associated factor (32%). Headache was reported in 98% followed by double vision (32%). Papilledema was present in all patients at the time of examination (100%). Perimetric study showed mild peripheral visual field constriction in 56%. Only two patients showed severe field constriction and one of them deteriorated rapidly and she became blind. The mean cerebrospinal fluid pressure was 302.5 mmH(2)O. The visual status improved significantly throughout the follow-up period. CONCLUSIONS: Idiopathic intracranial hypertension has relatively benign course in this part of the world and more aggressive treatment is not recommended. PMID- 16218912 TI - Sequential analyses of neurobiochemical markers of cerebral damage in cerebrospinal fluid and serum in CNS infections. AB - OBJECTIVE: To elucidate the relation between release patterns and cerebrospinal fluid/serum concentrations of neurobiochemical markers of cerebral damage and their potential value as monitoring parameters in central nervous system infections. METHODS: We investigated protein S-100B and neuron-specific enolase (NSE) in 102 sequential cerebrospinal fluid (CSF)-serum-pairs in patients with bacterial (n = 11) or viral (n = 13) meningitis/meningoencephalitis and neuroborreliosis (n = 8) in comparison with controls (n = 13). RESULTS: Highest S 100B values in CSF and serum were found on admission and showed a significant decrease afterwards. Comparison between disease groups revealed significant differences between bacterial and viral meningitis and neuroborreliosis for S 100B and also when compared with controls. NSE was not significantly elevated. CONCLUSIONS: S-100B is altered in CNS infection but does not provide additional benefit in the differential diagnosis when compared with standard CSF parameters. Nevertheless, S-100B values might be used as an additional monitoring parameter especially when sequential lumbar punctures are contraindicated. PMID- 16218913 TI - Impact of vertebral artery disease on dynamic cerebral autoregulation. AB - OBJECTIVES: This study applied dynamic cerebral autoregulation (DCA) testing distally to severe bilateral vertebral artery disease (BVAD). METHODS: Using continuous monitoring of beat-to-beat blood pressure and transcranial Doppler of the posterior cerebral arteries (PCA) were examined in 20 patients with BVAD and 22 controls. DCA testing was based on the 'high-pass filter model', which predicts a positive phase relationship between spontaneous oscillations (M-waves 3-9 cpm and R-waves 9-20 cpm) in blood pressure and cerebral blood flow velocity. RESULTS: In patients with BVAD, DCA testing detected autoregulatory deficits of different degrees. The lowest M-wave phase shift angles were found in the PCA territory distally to intracranial BVAD. CONCLUSION: This study suggests that DCA testing of the PCA could help to quantify the hemodynamic impact of BVAD. It highlights the relevance of functional TCD sonography as a useful diagnostic tool for the hemodynamic evaluation of vertebrobasilar disease. PMID- 16218914 TI - Mediterranean and Amerindian MHC class II alleles are associated with multiple sclerosis in Mexicans. AB - OBJECTIVES: Human leukocyte antigen (HLA)-DRB1, DQA1, DQB1 allele typing was performed in Mexicans Mestizos with multiple sclerosis (MS) to define the HLA class II alleles associated with the disease in this population. METHODS: Patients (n = 51) diagnosed according to the Poser criteria and a group of 173 unrelated healthy subjects were studied. PCR-SSOP and PCR-SSP were used for genotyping. RESULTS: Fifty five percent of the patients were females. The mean age at disease onset was 27 years. A relapsing-remitting disease was the most frequent type of MS (67%). A significant association of DRB1*0403 (OR = 5.68) with MS was shown. DRB1*0802 was also involved in susceptibility (OR = 2.41). An excess of DRB1*0802 homozygotes was observed in patients (P = 0.005), this genotype being in genetic equilibrium in controls. CONCLUSIONS: Two novel class II associations are described in Mexicans with MS: DRB1*0403 and DRB1*0802. Both alleles share with DRB1*1501, valine-86 and negatively charged amino acids, in the DRB1-anchoring motif of pocket 4. PMID- 16218915 TI - Is inadequate family history a barrier to diagnosis in CADASIL? AB - OBJECTIVES: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) has typical clinical features that include stroke, migraine, mood disturbances and cognitive decline. However, misdiagnosis is common. We hypothesized that family history is poorly elicited in individuals presenting with features of CADASIL and that enquiry into family history of all four cardinal manifestations of CADASIL is superior to elicitation of family history of premature stroke alone in raising the diagnostic possibility of CADASIL. MATERIALS AND METHODS: Retrospective review of family histories at presentation in 40 individuals with confirmed CADASIL was performed through structured interview in a Neurovascular Genetics clinic (182 first-degree and 242 second-degree relatives identified). Family history obtained from structured interview was compared to family history initially documented at presentation. RESULTS: At initial presentation, 30% of individuals were inaccurately documented to have no family history of significant neurological illness. Thirty-five per cent of patients had an initial alternative diagnosis. Initial inaccurate documentation of negative family history was more frequent in individuals with an initial alternative diagnosis. After structured interviews, 34% of 182 first degree and 35% of 242 second-degree relatives of CADASIL patients had history of stroke (16% of first-degree relatives had stroke before the age of 50 years). Forty-three per cent of first-degree and 28% of second-degree relatives had migraine, mood disturbance or cognitive decline. CONCLUSIONS: A false-negative family history was commonly documented in individuals presenting with features of CADASIL and was associated with initial misdiagnosis. Restriction of family history to premature stroke alone is probably inadequate to identify affected CADASIL pedigrees. PMID- 16218916 TI - Relationship between microtubule-binding repeats and morphology of neurofibrillary tangle in Alzheimer's disease. AB - OBJECTIVE: The present study was performed to compare the distributions of three repeat (3R) and four-repeat (4R) neurofibrillary tangles (NFT) with those of pretangles (p-NFT), intracellular NFT (i-NFT), and extracellular NFT (e-NFT) in the hippocampus of Alzheimer's disease brains. METHODS: NFT labeling was performed using anti-tau antibodies: pSer262 for p-NFT, pSer422 for i-NFT, AT8 for e-NFT, RD3 for 3R, and RD4 for 4R tau, and Gallyas impregnation for the NFT population. RD4- and pSer422-positive NFT were detected predominantly in sectors from CA2 to CA4, while RD3- and pSer262-positive NFT were predominantly present in CA1, the entorhinal cortex, and the subiculum. The tau epitope recognized by pSer262 belongs to 4R tau but it showed a strong correlation with RD3- and AT8 positive NFT. CONCLUSIONS: Sectors CA2-CA4 showed predominantly 4R-NFT containing the pSer422 epitope. pSer262 may detect the process of transformation from p-NFT to i-NFT, and e-NFT consisted predominantly of 3R tau. PMID- 16218917 TI - Reduction in head size in patients with aspartylglucosaminuria. AB - OBJECTIVE: To show that the head may shrink in adult patients with aspartylglucosaminuria (AGU), a neurodegenerative disease. METHOD: The head circumference (HC) of 40 adult patients (age at baseline 15 to 47) was measured twice with an interval of 10 years. Of these 40, 21 aged 15-47 and 19 young patients aged 5-14 as well as 40 healthy controls underwent lateral cephalometric radiography. RESULTS: During 10 years' follow-up, the HC of 26 (65%) had decreased by 1 to 4.5 cm (mean 1.7, P < 0.001). Evaluation of lateral skull radiographs revealed that patients aged 15 or more had significantly thicker skulls than did younger patients (P = 0.015). Mean intracranial length (glabella opisthocranium) of the patients aged 15 or more was significantly shorter than in patients aged 14 years or less (P = 0.029). These measurements indicated that brain volume had decreased. CONCLUSIONS: Macrocephalia in childhood followed by reduced brain volume in adulthood is evident in patients with AGU and is reflected by a decrease in head size. PMID- 16218918 TI - Middle age cognition and vocational outcome of childhood brain injury. AB - OBJECTIVES: In this almost four-decade follow-up we studied the cognitive performance of a cohort of 22 traumatic brain injury (TBI) patients in relation to vocational outcome. The patients had suffered a moderate to severe TBI in traffic accidents as preschoolers. METHODS: The neuropsychological assessment included the Profile of Mood States questionnaire and the Neurobehavioral Rating Scale. The cognitive performance of full-time working patients was compared with that of those not working. RESULTS: Full-time working patients had significantly better intellectual performance than the patients not at work. Memory performance was partly defective in both groups but neither group had subjective memory complaints. All patients working full time lived in a marital relationship and had less neurobehavioral problems than the patients not at work. CONCLUSIONS: Good intellectual capacity, verbal memory, and marital status were connected with a positive outcome. We suggest that as late as in middle age in spite of moderate to severe childhood TBI, it is still possible for a subgroup of patients to live a normal productive life. PMID- 16218919 TI - Spinal subdural Staphylococcus aureus abscess: case report and review of the literature. AB - Spinal subdural abscesses (SSA) are rare and to date only 57 cases have been reported. The exact incidence of the SSA is unknown. The most affected region is the thoraco-lumbar spine and the most common bacterial source is Staphylococcus aureus. The timing for magnetic resonance imaging is very important in these patients. Because early diagnosis and emergent treatment is vital to prevent the formation or progression of neurologic deficits. PMID- 16218920 TI - Enhancement of the human inspiratory pressure-flow relationship via the stretch shortening cycle. PMID- 16218921 TI - Force-frequency relation and sodium-calcium exchange: moving the staircase. PMID- 16218922 TI - Arnold Heller and the lymph pump. AB - This article reviews studies on lymph propulsion in the lymph vessels by active contraction of the vessels, first described by Arnold Heller in 1869 in German language, and here translated into English. His observations were first confirmed by Beatrice Carrier (1926) and Howard Flory et al. (1927), and several groups were active up to World war II. Few publications appeared in the period 1940- 1960, followed by increasing activity and development of new experimental techniques for use both in various experimental animals and in humans. Recently it has been shown that passive lymph flow may add to active propulsion. Both mechanisms depend on lymph formation, i.e. the uptake of interstitial fluid by the initial lymph vessels which is still not well understood. PMID- 16218923 TI - Left ventricular pressure-volume relationships during normal growth and development in the adult rat--studies in 8- and 50-week-old male Wistar rats. AB - AIMS: Left ventricular (LV) pressure-volume relations provide relatively load independent indexes of systolic and diastolic LV function, but few data are available on pressure-volume relations during growth and development in the normal adult heart. Furthermore, to quantify intrinsic ventricular function the indexes should be normalized for heart weight. However, in many studies the indexes are reported in absolute terms, or body weight-correction is used as a surrogate for heart weight-correction. METHODS: We determined pressure-volume relations in young (8-week-old, n = 13) and middle-aged (50-week-old, n = 19) male Wistar rats in relation to their heart and body weights. The animals were anaesthetized and a 2F pressure-conductance catheter was introduced into the LV to measure pressure-volume relations. RESULTS: Heart and body weights were significantly higher in the 50-week-old rats, whereas the heart-to-body weight ratio was significantly lower (2.74 +/- 0.32 vs. 4.41 +/- 0.37 mg g(-1), P < 0.001). Intrinsic systolic function, quantified by the slopes of the end-systolic pressure-volume relation (E(ES)), the dP/dt(MAX) vs. end-diastolic volume relation (S-dP), and the preload recruitable stroke work relation (PRSW), normalized for heart weight, was slightly decreased in the 50-week-old rats (S dP: -6%, P < 0.004; PRSW: -3%, P < 0.06). Heart weight-corrected diastolic indexes were not significant different. The absolute indexes qualitatively showed the same results, but body-weight corrected pressure-volume indexes showed improved systolic function and significantly depressed diastolic function. CONCLUSIONS: Intrinsic systolic function slightly decreases from the juvenile to the middle-aged period in normal male Wistar rats. Furthermore, correction of pressure-volume indexes for body weight is not an adequate surrogate for heart weight-correction in these animals. PMID- 16218924 TI - Force-frequency relation in frog-ventricle is dependent on the direction of sodium/calcium exchange in diastole. AB - AIM: Force of contraction increases with stimulus-frequency in mammalian and amphibian hearts under control conditions. Here, we have analysed the mechanism of the force-frequency relation (FFR) in frog-ventricle. METHODS: Circular strips of frog-ventricle were subjected to field-stimulation with frequencies in the range 0.03-0.2 Hz and force recorded on a chart-recorder. In another protocol, varying rest-periods were imposed while the preparation beat steadily at 0.2 Hz and the effect of rest on post-rest beat amplitude was noted. RESULTS: Under control conditions, a positive FFR and a rest-induced decay of contraction amplitude (RID) were seen in the frequency range 0.03-0.2 Hz. With cadmium, nifedipine, nickel (40 micromol L(-1)), ryanodine and adrenaline (all drugs at 10 micromol L(-1) concentration, except nickel), the positive FFR and RID seen under control conditions persisted. When the bathing solution contained ouabain (10 micromol L(-1)) or low external sodium (40 mmol L(-1)), or high external calcium (5 mmol L(-1)), the FFR turned negative in the frequency range stated above and there were rest-induced potentiations (RIP). CONCLUSION: When the conditions favour a net leak of calcium in diastole from intracellular stores via the calcium-extrusive mode of sodium-calcium exchanger (NCX), FFR is positive. An increase in frequency lessens the diastolic interval and therefore the diastolic calcium leak, thereby augmenting force. On the other hand, interventions which favour the calcium-acquisitive mode of NCX during diastole, changed the pattern of RID to RIP and converted FFR from positive to negative. With net diastolic calcium uptake, there is better store-filling and therefore higher force at lower frequencies. PMID- 16218925 TI - Effect of carbohydrate ingestion on brain exchange of amino acids during sustained exercise in human subjects. AB - AIM: This study investigated the effect of prolonged exercise with and without carbohydrate intake on the brain exchange of amino acids, especially focussing on tryptophan and branched-chain amino acids (BCAA). METHODS: Five male subjects exercised for 3 h on a cycle ergometer at 200 +/- 7 W on two occasions; either supplemented with a 6% carbohydrate solution or with flavoured water (placebo). Catheters were inserted into the right internal jugular vein and the radial artery of the non-dominant arm. The brain exchange of amino acids during exercise was calculated from the arterial-jugular venous concentration difference multiplied by plasma flow. RESULTS: About 106 micromol (22 mg) of tryptophan was taken up by the brain during exercise in the placebo trial, whereas no significant uptake was observed in the carbohydrate trial. In accordance, the arterial concentration of free tryptophan increased from 12 +/- 1 to 20 +/- 2 micromol L(-1) during the placebo trial and was significantly higher compared with the glucose trial (14 +/- 1 micromol L(-1) at the end of exercise). Also, the arterial concentration of total tryptophan (free and albumin-bound) increased during the first 30 min of exercise in both trials, but returned to the basal level at 180 min of exercise. In both trials, BCAA were taken up by the brain while glutamine was released. CONCLUSION: The present data show that both tryptophan and BCAA are taken up by the brain during prolonged exercise, and we suggest that the cerebral uptake of tryptophan may relate to increased synthesis of serotonin (5-HT) in the brain. PMID- 16218926 TI - Oxidative stress and nitric oxide synthase in skeletal muscles of rats with post infarction, compensated chronic heart failure. AB - AIM: Involvement of oxidative stress and nitric oxide synthase (NOS) isoforms in skeletal muscle cellular adaptations to chronic heart failure (CHF) is controversial, and possible muscle fibre-type heterogeneity in the oxidative stress and NOS responses to CHF have not been examined. Consequently, we hypothesized that the changes in determinants of elevated oxidative and nitrosylative stress associated with CHF would occur in skeletal muscle and would be similar in predominantly type I slow twitch muscle (soleus) and type II fast twitch muscle (plantaris) of rats. METHODS: The purpose of this study was to measure NOS isoforms (endothelial, inducible and neuronal NOS) and antioxidant enzymes (SOD-1, SOD-2, catalase) by protein immunoblot as well as markers of oxidative stress by biochemical assays in soleus and plantaris muscle sections of the rat hind limb. This was performed for control and post-infarction, compensated CHF rats. RESULTS: Twelve weeks after coronary artery ligation induced moderate CHF, soleus exhibited decreased SOD-1, SOD-2 and eNOS, but increased iNOS and nNOS isoforms assessed by immunoblot. This was associated with elevated lipid and DNA oxidative damage assessed by biochemical assays. In contrast, plantaris muscle exhibited no changes in antioxidant enzymes or NOS isoforms, and had lower lipid and DNA oxidative damage. CONCLUSION: These observations suggest a heretofore unreported muscle fibre-type-specific response of oxidative stress and NOS isoforms to CHF is of importance in understanding the cellular mechanisms of skeletal muscle dysfunction in CHF. PMID- 16218927 TI - Effects of innervation state on Hsp25 content and phosphorylation in inactive rat plantaris muscles. AB - AIM: Previous reports suggest a role for neuromuscular activity levels and/or connectivity in modulating Hsp25 expression and phosphorylation (pHsp25) in skeletal muscles. However, pHsp25 has only been studied in denervated muscles and/or muscles exposed to high levels of residual neuromuscular activity. Spinal cord isolation (SI) provides a model in which the muscle is exposed to nearly complete inactivity with maintenance of the nerve-muscle connection. To parcel out the roles of innervation state and activity-independent neural factors, we compared Hsp25 and pHsp25 in the plantaris of control (Con), SI, and denervated (Den, inactivity without neural connectivity) rats. METHODS: Hsp25 and pHsp25 protein levels (soluble and insoluble fractions) were measured with Western blot analysis after 1, 3, 8, 14, or 28 days of SI or Den. pHsp25 was normalized to non pHsp25 at each time point. RESULTS: Hsp25 was unchanged (days 1, 3 and 14) or increased (days 8 and 28) in the soluble fraction, and decreased (day 1) or increased (days 3, 8 and 14) in the insoluble fraction in Den compared with Con rats. pHsp25 was reduced after 1 and 28 days of Den, but near control levels on days 3, 8, and 14 in the soluble fraction. In the insoluble fraction, pHsp25 levels were lower in Den than Con rats on all days. In both fractions, Hsp25 was lower in SI than Con rats. pHsp25 levels were lower in the soluble fraction and higher in the insoluble fraction in SI than Con rats. CONCLUSION: These results suggest that an intact innervation, even in the absence of muscle activation and/or loading, is critical for Hsp25 phosphorylation in the insoluble fraction. However, the time-dependent decrease in Hsp25 with SI suggests a role for minimal levels of muscle activation and/or loading in maintaining Hsp25 expression during sustained inactivity. PMID- 16218928 TI - Mitochondrial efficiency in rat skeletal muscle: influence of respiration rate, substrate and muscle type. AB - AIM: To investigate the hypothesis that mitochondrial efficiency (i.e. P/O ratio) is higher in type I than in type II fibres during submaximal rates of respiration. METHODS: Mitochondria were isolated from rat soleus and extensor digitorum longus (EDL) muscles, representing type I and type II fibres, respectively. Mitochondrial efficiency (P/O ratio) was determined with pyruvate (Pyr) or palmitoyl-l-carnitine (PC) during submaximal (constant rate of adenosine diphosphate infusion) and maximal (V(max), state 3) rates of respiration and fitted to monoexponential functions. RESULTS: There was no difference in V(max) between PC and Pyr in soleus but in EDL V(max) with PC was only 58% of that with Pyr. The activity of 3-hydroxyacyl-CoA dehydrogenase was threefold higher in soleus than in EDL. P/O ratio at V(max) was 8-9% lower with PC [2.33 +/- 0.02 (soleus) and 2.30 +/- 0.02 (EDL)] than with Pyr [2.52 +/- 0.03 (soleus) and 2.54 +/- 0.03 (EDL)] but not different between the two muscles (P > 0.05). P/O ratio was low at low rates of respiration and increased exponentially when the rate of respiration increased. The asymptotes of the curves were similar to P/O ratio at V(max). P/O ratio at submaximal respirations was not different between soleus and EDL neither with Pyr nor with PC. CONCLUSION: Mitochondrial efficiency, as determined in vitro, was not significantly different in the two fibre types neither at V(max) nor at submaximal rates of respiration. The low V(max) for PC oxidation in EDL may relate to low activity of beta-oxidation. PMID- 16218929 TI - Saline-induced natriuresis and renal blood flow in conscious dogs: effects of sodium infusion rate and concentration. AB - AIM: This study focused on static and dynamic changes in total renal blood flow (RBF) during volume expansion and tested whether a change in RBF characteristics is a necessary effector mechanism in saline-induced natriuresis. METHODS: The aortic flow subtraction technique was used to measure RBF continuously. Identical amounts of NaCl (2.4 mmol kg(-1)) were given as slow isotonic (Iso, 120 min), slow hypertonic (Hyper, 120 min), and rapid isotonic loads (IsoRapid, 30 min). RESULTS: During Iso and IsoRapid, arterial blood pressure increased slightly (6-7 mmHg), and during Hyper it remained unchanged. Iso and Hyper increased sodium excretion (4 +/- 1 to 57 +/- 27 and 10 +/- 4 to 79 +/- 28 micromol min(-1), respectively) and decreased plasma renin activity (by 38% and 29%), angiotensin II (by 56% and 58%) and aldosterone (by 47% and 65%), while RBF remained unchanged. IsoRapid caused a similar increase in sodium excretion (to 72 +/- 19 micromol min(-1)), a similar decrease in renin system activity, but a 15% elevation of RBF (282 +/- 22 to 324 +/- 35 mL min(-1)). Selected frequency domain parameters of RBF autoregulation did not change in response to any load. CONCLUSIONS: In response to slow saline loading simulating daily sodium intake, the rate of sodium excretion may increase 10-20-fold without any change in mean arterial blood pressure or in RBF. Regulatory responses to changes in total body NaCl levels appears, therefore, to be mediated primarily by neurohumoral mechanisms and may occur independent of changes in arterial pressure or RBF. PMID- 16218930 TI - Respiratory muscle performance with stretch-shortening cycle manoeuvres: maximal inspiratory pressure-flow curves. AB - AIM: To test the hypothesis that the maximal inspiratory muscle (IM) performance, as assessed by the maximal IM pressure-flow relationship, is enhanced with the stretch-shortening cycle (SSC). METHODS: Maximal inspiratory flow-pressure curves were measured in 12 healthy volunteers (35 +/- 6 years) during maximal single efforts through a range of graded resistors (4-, 6-, and 8-mm diameter orifices), against an occluded airway, and with a minimal load (wide-open resistor). Maximal inspiratory efforts were initiated at a volume near residual lung volume (RV). The subjects exhaled to RV using slow (S) or fast (F) manoeuvres. With the S manoeuvre, they exhaled slowly to RV and held the breath at RV for about 4 s prior to maximal inspiration. With the F manoeuvre, they exhaled rapidly to RV and immediately inhaled maximally without a post-expiratory hold; a strategy designed to enhance inspiratory pressure via the SSC. RESULTS: The maximal inspiratory pressure-flow relationship was linear with the S and F manoeuvres (r2 = 0.88 for S and r2 = 0.88 for F manoeuvre, P < 0.0005 in all subjects). With the F manoeuvre, the pressure-flow relationship shifted to the right in a parallel fashion and the calculated maximal power increased by approximately 10% (P < 0.05) over that calculated with the S manoeuvre. CONCLUSION: The maximal inspiratory pressure-flow capacity can be enhanced with SSC manoeuvres in a manner analogous to increases in the force-velocity relationship with SSC reported for skeletal muscles. PMID- 16218931 TI - Composite glandular-endocrine cell carcinomas of the stomach: clinicopathologic and methylation study. AB - Four cases of very rare composite glandular-endocrine cell carcinoma of the stomach are presented with methylation findings. All but one of the tumors arose in the antrum and two of them were at the early stage. Each composite carcinoma was accompanied by atrophic and metaplastic gastritis in the adjacent mucosa. Three cases showed lymph nodes metastasis, and one of them showed both glandular and neuroendocrine tumor components within the metastatic nodes. Mucin stains were positive in the adenocarcinoma areas while only the neuroendocrine markers were positive in neuroendocrine tumor components. Of all seven markers tested for, p16INK4A methylation was observed in both components of one composite carcinoma and hMLH1 was methylated in the neuroendocrine tumor component within the same tumor. An additional six gastric large cell neuroendocrine carcinomas showed no methylation. Follow up of patients indicated short survival in patients with poorly differentiated neuroendocrine carcinoma components and advanced stages of tumors, while patients with well-differentiated neuroendocrine tumor components and early stages showed long disease-free survival. Our results suggest that hypermethylation of tumor suppressor genes is rare in gastric composite and neuroendocrine carcinomas, and prognosis of gastric composite carcinomas appears to be related to the histopathology of neuroendocrine components and tumor stage. PMID- 16218932 TI - Virulence genes in verocytotoxigenic Escherichia coli strains isolated from humans and cattle. AB - Verocytotoxigenic Escherichia coli (VTEC) causing diarrhoea, haemorrhagic colitis and haemolytic-uremic syndrome usually have additional traits such as the adhesin intimin and a large plasmid that seems to increase virulence. There are, however, isolates of VTEC causing serious symptoms that do not harbour these traits. In the present study we have used PCR with primers detecting adhesin genes other than eaeA, namely fimA, papC, sfaD/sfaE and daaE. We have also used PCR to detect the genes hlyA and iutA that besides the plasmid-borne gene E-hly possibly support the bacterial access to iron. The aim of the study was to identify and compare the presence of virulence genes in VTEC isolates of human and cattle origin. The main finding was that the absence of E-hly might be compensated for by the gene iutA coding for aerobactin or hlyA coding for alpha-haemolysin as 94% of the human VTEC isolates had at least one of these genes. Interestingly, only 45% of VTEC isolated from cattle had any of these genes. We propose that this might be the reason for the relatively low incidence of symptomatic VTEC infections among humans in relation to the high number of VTEC among cattle. PMID- 16218933 TI - Improved diagnosis of mycobacterial infections in formalin-fixed and paraffin embedded sections with nested polymerase chain reaction. AB - Traditional histological diagnosis of mycobacterial infection in formalin-fixed and paraffin-embedded (FFPE) tissues is insensitive and poorly specific. To improve this, we developed nested polymerase chain reaction (PCR) protocols for detecting a Mycobacterium genus-specific 65-kDa heat shock protein (HSP65) sequence and the M. tuberculosis complex-specific insertion sequence IS6110 in FFPE sections. Protocols were optimized on tissues from 20 patients with a final clinical diagnosis of mycobacterial infection. Amplicons were controlled by sequencing and restriction endonuclease digestion. PCR could detect as few as three mycobacterial genomes per reaction. Assays showed 100% sensitivity and specificity for both M. tuberculosis complex and M. avium complex infection. Paraffin blocks from a second group of 26 patients with histological evidence of necrotizing granulomas of unknown etiology were then analyzed as a surrogate group to test the assay under conditions similar to those applying during routine diagnosis. Twenty-three of these blocks contained amplifiable DNA; nine were positive for M. tuberculosis complex DNA and four for other types of mycobacterial DNA. Furthermore, digestion of HSP65 amplicons with NarI could distinguish M. tuberculosis from M. avium complex. In conclusion, our nested PCR assays can be used as reliable tools for the detection of mycobacterial infections in FFPE tissues. The assays are simple and rapid to perform and show improved sensitivity and specificity compared to previously reported protocols. PMID- 16218934 TI - Cell-type-specific expression of p53 and p21 in giant cell arteritis. AB - The aim of the present study was to investigate the expression of TP53 (p53) and CDKN1A (CIP1; p21) in the arterial wall in giant cell arteritis (GCA). Cross sections from 18 temporal artery biopsies displaying GCA and 8 control arteries were double-stained with monoclonal antibody directed at p53 or p21 on the one hand and alpha-smooth muscle actin, CD68 (macrophage) or CD3 (T-cell) on the other. Nuclear p53 was expressed in CD68-positive cells and smooth muscle cells in 16 of the 18 inflamed arteries. P21-positive nuclei were found in CD68 positive cells in 14 biopsies and in smooth muscle cells in all the specimens. All p53-positive giant cells also contained p21-positive nuclei. In the giant cells, immunopositive nuclei were mixed with negative ones. CD3-positive T-cells did not express p53 or p21. Only one p53-positive smooth muscle cell nucleus was found in the non-GCA controls and, compared with GCA, p21 expression was noted in few smooth muscle nuclei. The presence of p53 and p21 in the same types of cell in GCA indicates that the former protein is functional; p21 expression is induced by wild-type, functional p53 but not by its mutant form. The current observations suggest cellular stress in GCA, the nature of which requires further investigation. PMID- 16218935 TI - Immunohistochemical expression of thymidylate synthase as predictor of response to capecitabine in patients with advanced colorectal adenocarcinoma. AB - Capecitabine is an oral prodrug to 5-fluorouracil (5-FU). The primary target of 5 FU is thymidylate synthase (TS). A mainstay of colorectal adenocarcinoma chemotherapy is inhibition of TS, which may be one of many determinant factors when predicting the outcome of chemotherapies based on fluoropyrimidine treatment. This retrospective study included 39 patients with advanced colorectal adenocarcinoma treated with capecitabine. Response was assessed by measuring the amount of tumour in the course of treatment. TS expression was evaluated by scoring the immunohistochemical (IHC) reaction and assessing the predominant IHC reaction pattern. This study showed significant correlation between the predominant IHC reaction pattern and response, but no correlation between IHC score and response. The predominant IHC reaction pattern may be a useful parameter in prediction of clinical outcome in patients treated with capecitabine. PMID- 16218936 TI - Phene Plate (PhP) biochemical fingerprinting. A screening method for epidemiological typing of enterococcal isolates. AB - Pulsed-field gel electrophoresis (PFGE) is currently considered the gold standard for genotyping of enterococci. However, PFGE is both expensive and time consuming. The purpose of this study was to investigate whether the PhP system can be used as a reliable clinical screening method for detection of genetically related isolates of enterococci. If so, it should be possible to minimize the number of isolates subjected to PFGE typing, which would save time and money. Ninety-nine clinical enterococcal isolates were analysed by PhP (similarity levels 0.90-0.975) and PFGE (similarity levels < or =3 and < or =6 bands) and all possible pairs of isolates were cross-classified as matched or mismatched. We found that the probability that a pair of isolates (A and B) belonging to the same type according to PhP also belong to the same cluster according to PFGE, i.e. p(A(PFGE)=B(PFGE) * A(PhP)=B(PhP)), and the probability that a pair of isolates of different types according to PhP also belong to different clusters according to PFGE, i.e. p(A(PFGE) not equalB(PFGE) * A(PhP) not equalB(PhP)), was relatively high for E. faecalis (0.86 and 0.96, respectively), but was lower for E. faecium (0.51 and 0.77, respectively). The concordance which shows the probability that PhP and PFGE agree on match or mismatch was 86%-93% for E. faecalis and 54%-66% for E. faecium, which indicates that the PhP method may be useful for epidemiological typing of E. faecalis in the current settings but not for E. faecium. PMID- 16218938 TI - Ribosomal DNA sequencing: experiences from use in the Danish National Reference Laboratory for Identification of Bacteria. AB - Diagnostic tools for identification of bacteria have developed dramatically in the last decade. Sequencing of genes coding for rRNA has led to revolutionary insights into the phylogeny and taxonomy of bacteria, and to new demands on the service provided by national reference laboratories for identification of bacteria. At the Danish Reference Laboratory for Identification of Bacteria, partial 16S rDNA sequencing has been used since 2001 to identify "difficult" strains submitted for taxonomic elucidation. Experiences relating to phenotypic as well as 16S rDNA sequencing of the first 175 strains examined are presented. Approximately 2/3 of the strains were Gram-positive and 1/3 Gram-negative. One fifth of the strains were anaerobic, while 4/5 were either facultatively anaerobic or aerobic. Methodological agreement was seen for most strains at species and/or genus level. Methodological disagreement was relatively rare. In 1/6 of the strains valuable information was obtained from sequencing results, while for some strains identification was based primarily on the phenotypic results. Only a few strains could not be clearly identified by either method. A very large number of strains representing taxons ranging from facultatively anaerobic to aerobic and anaerobic species and genera, Gram-positive as well as Gram-negative, were successfully examined. Of the submitted strains many have only rarely been encountered as human pathogens. Thus, genotypic identification may result in recognition of hitherto seldom recognized or unrecognized bacteria as human pathogens, which will lead to a better understanding of the nature of human infections. It is self evident that we should focus on slowly growing, fastidious or 'difficult' organisms when using sequencing for national reference purposes. Short sequences (450-650 base pairs) seem sufficient for most identifications. Molecular bacterial identification is a powerful tool for national reference laboratories, enhancing both the speed and validity of examinations performed. PMID- 16218937 TI - Cloning and expression of rat fucosyltransferase VII at sites of inflammation. AB - The sialyl Lewis x (NeuAcalpha2-3Galbeta1-4(Fucalpha1-3)GlcNAc) determinants serve as ligands in the selectin-mediated adhesion of leukocytes to activated endothelium. The final step in the sialyl Lewis x synthesis is catalyzed by alpha1-3-fucosyltransferase, which transfers fucose to sialylated type 2 chain. We report the cloning of rat alpha1-3-fucosyltransferase gene (rFUT) isolated from rat lymph node and kidney allograft. The rFUT is expressed as two splice variants, but only the long one showed enzymatic activity towards sialylated lactosamine. Also flow cytometry analysis with the sLex mAbs indicated that the cloned rFuc-T was a functional enzyme and a member of the Fuc-TVII family. The rFuc-TVII mRNA expression level was strongly enhanced during acute inflammatory reaction induced by kidney allograft rejection, which could be detected by in situ hybridization and quantitative real-time PCR. PMID- 16218939 TI - Verruciform xanthoma of the oral cavity: clinicopathological study relating to pathogenesis. Report of three cases. AB - Verruciform xanthoma is a rare condition that was first reported in the oral cavity in 1971. Its histopathology is distinctive on account of the presence of foamy histiocytes within elongated dermal papillae. Three cases of oral mucosal verruciform xanthoma were studied. Immunohistochemical staining by streptavidin peroxidase and in situ hybridization to detect human papillomavirus (HPV types 6, 11, 16, 18) DNA and matrix metalloproteinase (MMP-2, -9) RNA were performed to investigate the pathogenesis of verruciform xanthoma. This study showed that the foam cells were strongly positive for CD68 (KP1) and vimentin. Cytokeratin, PCNA and S-100 stained focally negative in foam cells. In situ hybridization failed to detect HPV (types 6, 11, 16, 18) in any of the three cases. Based on our findings we conclude that verruciform xanthoma is most likely not a human papillomavirus associated lesion; the foam cells, as a histological hallmark of the lesion, are most likely derived from the monocyte-macrophage lineage, and verruciform xanthoma is, at least partly, mediated by an immune mechanism. MMPs degrade basilar membrane that promotes the reciprocal induction between epithelium and mesenchyme. However, as yet unrecognized factors may play a role in the development of epithelium-mesenchyme reciprocal induction. PMID- 16218940 TI - Statin-associated myopathy with normal creatine kinase levels. Case report from a Norwegian family. AB - Troseid M, Henriksen OA, Lindal S. Statin-associated myopathy with normal creatine kinase levels - case report from a Norwegian family. APMIS 2005;113:635 7. Recent reports suggest that statins may cause myopathy with normal creatine kinase levels. We describe four related patients with statin-associated muscle symptoms and normal creatine kinase levels. In two out of the four patients (mother and son), pathological findings on EMG suggested myopathy, and muscle biopsies showed evidence of mitochondrial pathology. A third patient (daughter) had slight myopathic findings on EMG and muscle biopsy, but not enough to be classified as pathological. In a fourth patient, there were no pathological findings. Creatine kinase levels were normal and symptoms diminished after discontinuation of drugs in all four patients. Our findings are consistent with other reports of statin-associated myopathy with normal creatine kinase levels. An inherited vulnerability, possibly a mitochondrial pathology, might cause or aggravate symptoms in some patients. PMID- 16218941 TI - Floral variant of follicular lymphoma containing marginal zone B-cell component. Report of two cases. AB - We here report two unusual cases of floral variant of follicular lymphoma containing marginal zone B-cells. Histologically, the neoplastic follicles consisted of three distinct layers. The inner layer was composed of neoplastic germinal centers exhibiting a floral design and the middle layer had unusually prominent mantle zones. The outer zone of neoplastic follicles was surrounded by a pale cuff of marginal zone B-cells. Immunohistological study demonstrated that both the germinal center and marginal zone component lay within the follicular dendritic cell network. The germinal center component was CD10+ and bcl-2+. However, a portion of the marginal zone component weakly expressed bcl-2 but not CD10. Nodal marginal zone B-cell lymphoma (NMZBL) occasionally possesses "floral" lymphoid follicles. Follicular lymphoma with marginal zone differentiation is a high-risk variant of follicular lymphoma. In diagnostic practice, the differential diagnosis between the floral variant of follicular lymphoma containing marginal zone B-cells and the "floral variant" of NMZBL is important. PMID- 16218942 TI - Malignant mullerian mixed tumor (carcinosarcoma) of the fallopian tube: an immunohistochemical study of neoplastic cells. AB - The patient was a 65-year-old woman who complained of lower abdominal pain. Salpingo-oophorectomy and hysterectomy were performed due to suspicion of ovarian cancer. At surgery a polypoid mass was observed in the fimbria of the left fallopian tube. Histologically, proliferation of undifferentiated neoplastic cells with marked cytological atypia predominated in the tumor. Proliferation of rhabdomyoblastic cells or spindle cells, as well as adenocarcinoma arising from the mucosa of the fallopian tube, was observed. A diagnosis of malignant mullerian mixed tumor (MMMT) was made. CD10 was expressed in adenocarcinoma, undifferentiated, spindle and rhabdomyoblastic cells. Furthermore, rhabdomyoblastic cells were positive for desmin and myoglobin. Undifferentiated and spindle neoplastic cells were focally positive for ASMA and negative for h caldesmon. Finally, our preliminary report suggests that MMMT of the fallopian tube may contain immature smooth muscle cells or cells with the myofibroblast like immunohistochemical phenotype in the undifferentiated component. PMID- 16218943 TI - Identifying protein interactions. PMID- 16218945 TI - Utilizing logical relationships in genomic data to decipher cellular processes. AB - The wealth of available genomic data has spawned a corresponding interest in computational methods that can impart biological meaning and context to these experiments. Traditional computational methods have drawn relationships between pairs of proteins or genes based on notions of equality or similarity between their patterns of occurrence or behavior. For example, two genes displaying similar variation in expression, over a number of experiments, may be predicted to be functionally related. We have introduced a natural extension of these approaches, instead identifying logical relationships involving triplets of proteins. Triplets provide for various discrete kinds of logic relationships, leading to detailed inferences about biological associations. For instance, a protein C might be encoded within an organism if, and only if, two other proteins A and B are also both encoded within the organism, thus suggesting that gene C is functionally related to genes A and B. The method has been applied fruitfully to both phylogenetic and microarray expression data, and has been used to associate logical combinations of protein activity with disease state phenotypes, revealing previously unknown ternary relationships among proteins, and illustrating the inherent complexities that arise in biological data. PMID- 16218944 TI - Protein database searches using compositionally adjusted substitution matrices. AB - Almost all protein database search methods use amino acid substitution matrices for scoring, optimizing, and assessing the statistical significance of sequence alignments. Much care and effort has therefore gone into constructing substitution matrices, and the quality of search results can depend strongly upon the choice of the proper matrix. A long-standing problem has been the comparison of sequences with biased amino acid compositions, for which standard substitution matrices are not optimal. To address this problem, we have recently developed a general procedure for transforming a standard matrix into one appropriate for the comparison of two sequences with arbitrary, and possibly differing compositions. Such adjusted matrices yield, on average, improved alignments and alignment scores when applied to the comparison of proteins with markedly biased compositions. Here we review the application of compositionally adjusted matrices and consider whether they may also be applied fruitfully to general purpose protein sequence database searches, in which related sequence pairs do not necessarily have strong compositional biases. Although it is not advisable to apply compositional adjustment indiscriminately, we describe several simple criteria under which invoking such adjustment is on average beneficial. In a typical database search, at least one of these criteria is satisfied by over half the related sequence pairs. Compositional substitution matrix adjustment is now available in NCBI's protein-protein version of blast. PMID- 16218946 TI - Identifying remote protein homologs by network propagation. AB - Perhaps the most widely used applications of bioinformatics are tools such as psi blast for searching sequence databases. We describe a recently developed protein database search algorithm called rankprop. rankprop relies upon a precomputed network of pairwise protein similarities. The algorithm performs a diffusion operation from a specified query protein across the protein similarity network. The resulting activation scores, assigned to each database protein, encode information about the global structure of the protein similarity network. This type of algorithm has a rich history in associationist psychology, artificial intelligence and web search. We describe the rankprop algorithm and its relatives, and we provide evidence that the algorithm successfully improves upon the rankings produced by psi-blast. PMID- 16218947 TI - Flexible nets. The roles of intrinsic disorder in protein interaction networks. AB - Proteins participate in complex sets of interactions that represent the mechanistic foundation for much of the physiology and function of the cell. These protein-protein interactions are organized into exquisitely complex networks. The architecture of protein-protein interaction networks was recently proposed to be scale-free, with most of the proteins having only one or two connections but with relatively fewer 'hubs' possessing tens, hundreds or more links. The high level of hub connectivity must somehow be reflected in protein structure. What structural quality of hub proteins enables them to interact with large numbers of diverse targets? One possibility would be to employ binding regions that have the ability to bind multiple, structurally diverse partners. This trait can be imparted by the incorporation of intrinsic disorder in one or both partners. To illustrate the value of such contributions, this review examines the roles of intrinsic disorder in protein network architecture. We show that there are three general ways that intrinsic disorder can contribute: First, intrinsic disorder can serve as the structural basis for hub protein promiscuity; secondly, intrinsically disordered proteins can bind to structured hub proteins; and thirdly, intrinsic disorder can provide flexible linkers between functional domains with the linkers enabling mechanisms that facilitate binding diversity. An important research direction will be to determine what fraction of protein protein interaction in regulatory networks relies on intrinsic disorder. PMID- 16218948 TI - Histones in functional diversification. Core histone variants. AB - Recent research suggests that minor changes in the primary sequence of the conserved histones may become major determinants for the chromatin structure regulating gene expression and other DNA-related processes. An analysis of the involvement of different core histone variants in different nuclear processes and the structure of different variant nucleosome cores shows that this may indeed be so. Histone variants may also be involved in demarcating functional regions of the chromatin. We discuss in this review why two of the four core histones show higher variation. A comparison of the status of variants in yeast with those from higher eukaryotes suggests that histone variants have evolved in synchrony with functional requirement of the cell. PMID- 16218949 TI - Dynamin-related proteins and Pex11 proteins in peroxisome division and proliferation. AB - The abundance and size of cellular organelles vary depending on the cell type and metabolic needs. Peroxisomes constitute a class of cellular organelles renowned for their ability to adapt to cellular and environmental conditions. Together with transcriptional regulators, two groups of peroxisomal proteins have a pronounced influence on peroxisome size and abundance. Pex11-type peroxisome proliferators are involved in the proliferation of peroxisomes, defined here as an increase in size and/or number of peroxisomes. Dynamin-related proteins have recently been suggested to be required for the scission of peroxisomal membranes. This review surveys the function of Pex11-type peroxisome proliferators and dynamin-related proteins in peroxisomal proliferation and division. PMID- 16218950 TI - The N-glycans of yellow jacket venom hyaluronidases and the protein sequence of its major isoform in Vespula vulgaris. AB - Hyaluronidase (E.C. 3.2.1.35), one of the three major allergens of yellow jacket venom, is a glycoprotein of 45 kDa that is largely responsible for the cross reactivity of wasp and bee venoms with sera of allergic patients. The asparagine linked carbohydrate often appears to constitute the common IgE-binding determinant. Using a combination of MALDI MS and HPLC of 2-aminopyridine-labelled glycans, we found core-difucosylated paucimannosidic glycans to be the major species in the 43-45 kDa band of Vespula vulgaris and also in the corresponding bands of venoms from five other wasp species (V. germanica, V. maculifrons, V. pensylvanica, V. flavopilosa and V. squamosa). Concomitant peptide mapping of the V. vulgaris 43 kDa band identified the known hyaluronidase, Ves v 2 (SwissProt P49370), but only as a minor component. De novo sequencing by tandem MS revealed the predominating peptides to resemble a different, yet homologous, sequence. cDNA cloning retrieved a sequence with 58 and 59% homology to the previously known isoform and to the Dolichovespula maculata and Polistes annularis hyaluronidases. Close homologues of this new, putative hyaluronidase b (Ves v 2b) were also the major isoform in the other wasp venoms. PMID- 16218951 TI - Transcription of individual tRNA1Gly genes from within a multigene family is regulated by transcription factor TFIIIB. AB - Members of a multigene family from the silkworm Bombyx mori have been classified based on their transcriptions in homologous nuclear extracts, into three groups of highly, moderately and poorly transcribed genes. Because all these gene copies have identical coding sequences and consequently identical promoter elements (the A and B boxes), the flanking sequences modulate their expression levels. Here we demonstrate the interaction of transcription factor TFIIIB with these genes and its role in regulating differential transcriptions. The binding of TFIIIB to the poorly transcribed gene -6,7 was less stable compared with binding of TFIIIB to the highly expressed copy, -1. The presence of a 5' upstream TATA sequence closer to the coding region in -6,7 suggested that the initial binding of TFIIIC to the A and B boxes sterically hindered anchoring of TFIIIB via direct interactions, leading to lower stability of TFIIIC-B-DNA complexes. Also, the multiple TATATAA sequences present in the flanking regions of this poorly transcribed gene successfully competed for TFIIIB reducing transcription. The transcription level could be enhanced to some extent by supplementation of TFIIIB but not by TATA box binding protein. The poor transcription of -6,7 was thus attributed both to the formation of a less stable transcription complex and the sequestration of TFIIIB. Availability of the transcription factor TFIIIB in excess could serve as a general mechanism to initiate transcription from all the individual members of the gene family as per the developmental needs within the tissue. PMID- 16218952 TI - Coenzyme A affects firefly luciferase luminescence because it acts as a substrate and not as an allosteric effector. AB - The effect of CoA on the characteristic light decay of the firefly luciferase catalysed bioluminescence reaction was studied. At least part of the light decay is due to the luciferase catalysed formation of dehydroluciferyl-adenylate (L AMP), a by-product that results from oxidation of luciferyl-adenylate (LH2-AMP), and is a powerful inhibitor of the bioluminescence reaction (IC50 = 6 nm). We have shown that the CoA induced stabilization of light emission does not result from an allosteric effect but is due to the thiolytic reaction between CoA and L AMP, which gives rise to dehydroluciferyl-CoA (L-CoA), a much less powerful inhibitor (IC50 = 5 microm). Moreover, the V(max) for L-CoA formation was determined as 160 min(-1), which is one order of magnitude higher than the V(max) of the bioluminescence reaction. Results obtained with CoA analogues also support the thiolytic reaction mechanism: CoA analogues without the thiol group (dethio CoA and acetyl-CoA) do not react with L-AMP and do not antagonize its inhibitor effect; CoA and dephospho-CoA have free thiol groups, both react with L-AMP and both antagonize its effect. In the case of dephospho-CoA, it was shown that it reacts with L-AMP forming dehydroluciferyl-dephospho-CoA. Its slower reactivity towards L-AMP explains its lower potency as antagonist of the inhibitory effect of L-AMP on the light reaction. Moreover, our results support the conjecture that, in the bioluminescence reaction, the fraction of LH2-AMP that is oxidized into L-AMP, relative to other inhibitory products or intermediates, increases when the concentrations of the substrates ATP and luciferin increases. PMID- 16218953 TI - Analysis of the contribution of changes in mRNA stability to the changes in steady-state levels of cyclin mRNA in the mammalian cell cycle. AB - Cyclins are the essential regulatory subunits of cyclin-dependent protein kinases. They accumulate and disappear periodically at specific phases of the cell cycle. Here we investigated whether variations in cyclin mRNA levels in exponentially growing cells can be attributed to changes in mRNA stability. Mouse EL4 lymphoma cells and 3T3 fibroblasts were synchronized by elutriation or cell sorting. Steady-state levels and degradation of cyclin mRNAs and some other cell cycle related mRNAs were measured at early G1, late G1, S and G2/M phases. In both cell lines mRNAs of cyclins C, D1 and D3 remained unchanged throughout the cell cycle. In contrast, cyclin A2 and B1 mRNAs accumulated 3.1- and 5.7-fold between early G1 and G2/M phase, whereas cyclin E1 mRNA decreased 1.7-fold. Mouse cyclin A2 and B1 genes, by alternative polyadenylation, gave rise to more than one transcript. In both cases, the longer transcripts were the minor species but accumulated more strongly in G2/M phase. All mRNAs were rather stable with half lives of 1.5-2 h for cyclin E1 mRNA and 3-4 h for the others. Changes in mRNA stability accounted for the accumulation in G2/M phase of the short cyclin A2 and B1 mRNAs, but contributed only partially to changes in levels of the other mRNAs. PMID- 16218954 TI - Characterization of novel sequence motifs within N- and C-terminal extensions of p26, a small heat shock protein from Artemia franciscana. AB - The small heat shock proteins function as molecular chaperones, an activity often requiring reversible oligomerization and which protects against irreversible protein denaturation. An abundantly produced small heat shock protein termed p26 is thought to contribute to the remarkable stress resistance exhibited by encysted embryos of the crustacean, Artemia franciscana. Three novel sequence motifs termed G, R and TS were individually deleted from p26 by site-directed mutagenesis. G encompasses residues G8-G29, a glycine-enriched region, and R includes residues R36-R45, an arginine-enhanced sequence, both in the amino terminus. TS, composed of residues T169-T186, resides in the carboxy-extension and is augmented in threonine and serine. Deletion of R had more influence than removal of G on p26 oligomerization and chaperoning, the latter determined by thermotolerance induction in Escherichia coli, protection of insulin and citrate synthase from dithiothreitol- and heat-induced aggregation, respectively, and preservation of citrate synthase activity upon heating. Oligomerization of the TS and R variants was similar, but the TS deletion was slightly more effective than R as a chaperone. The extent of p26 structural perturbation introduced by internal deletions, including modification of intrinsic fluorescence, 1-anilino-8 naphthalene-sulphonate binding and secondary structure, paralleled reductions in oligomerization and chaperoning. Three-dimensional modeling of p26 based on wheat Hsp16.9 crystal structure indicated many similarities between the two proteins, including peptide loops associated with secondary structure elements. Loop 1 of p26 was deleted in the G variant with minimal effect on oligomerization and chaperoning, whereas loop 3, containing beta-strand 6 was smaller than the corresponding loop in Hsp16.9, which may influence p26 function. PMID- 16218955 TI - Inhibition of Hsp90 function delays and impairs recovery from heat shock. AB - The induction of the heat shock response as well as its termination is autoregulated by heat shock protein activities. In this study we have investigated whether Hsp90 functional protein levels influence the characteristics and duration of the heat shock response. Treatment of cells with several benzoquinone ansamycin inhibitors of Hsp90 (geldanamycin, herbimycin A) activated a heat shock response in the absence of heat shock, as reported previously. Pretreatment of cells with the Hsp90 inhibitors significantly delayed the rate of restoration of normal protein synthesis following a brief heat shock. Concurrently, the rate of Hsp synthesis and accumulation was substantially increased and prolonged. The cessation of heat shock protein synthesis did not occur until the levels of Hsp70 were substantially elevated relative to its standard threshold for autoregulation. The elevated levels of HSPS 22-28 (the small HSPS) and Hsp70 are not able to promote thermotolerance when Hsp90 activity is repressed by ansamycins; rather a suppression of thermotolerance is observed. These results suggest that a multicomponent protein chaperone complex involving both Hsp90 and Hsp70 signals the cessation of heat shock protein synthesis, the restoration of normal translation, and likely the establishment of thermotolerance. Impaired function of either component is sufficient to alter the heat shock response. PMID- 16218956 TI - Two different types of hepcidins from the Japanese flounder Paralichthys olivaceus. AB - The cysteine-rich peptide hepcidin is known to be an antimicrobial peptide and iron transport regulator that has been found in both fish and mammals. Recently, we found two different types (designated Hep-JF1 and Hep-JF2) of hepcidin cDNA in the Japanese flounder, Paralichthys olivaceus, by expressed sequence tag analysis. The identity of amino acid sequences between Hep-JF1 and Hep-JF2 was 51%. The Hep-JF1 and Hep-JF2 genes both consist of three exons and two introns, and both exist as single copies in the genome. The predicted mature regions of Hep-JF1 and Hep-JF2 have six and eight Cys residues, respectively. The first Cys residue of Hep-JF1 was deleted and the second was replaced with Gly. The number and positions of Cys residues in Hep-JF2 are the same as they are in human Hep. Hep-JF1 is specifically expressed in liver while the expression of Hep-JF2 was detected from gill, liver, heart, kidney, peripheral blood leucocytes, spleen and stomach. Gene expression of Hep-JF1 in liver decreased during experimental iron (iron-dextran) overload. Expression of Hep-JF1 in liver was decreased by injecting fish with iron-dextran and increased by injecting lipopolysaccharide. Iron overload did not significantly affect expression of Hep-JF2 in liver but it did increase expression of Hep-JF2 in kidney. Lipopolysaccharide injection increased expression of Hep-JF2 in both liver and kidney. In liver, some cells expressed both Hep-JF1 and Hep-JF2 while some other cells expressed just one of them. Synthesized Hep-JF2 peptide showed antimicrobial activity, while synthesized Hep-JF1 peptide did not against several bacteria including fish pathogenic bacteria used in this study. PMID- 16218958 TI - Adenine and adenosine salvage pathways in erythrocytes and the role of S adenosylhomocysteine hydrolase. A theoretical study using elementary flux modes. AB - This article is devoted to the study of redundancy and yield of salvage pathways in human erythrocytes. These cells are not able to synthesize ATP de novo. However, the salvage (recycling) of certain nucleosides or bases to give nucleotide triphosphates is operative. As the salvage pathways use enzymes consuming ATP as well as enzymes producing ATP, it is not easy to see whether a net synthesis of ATP is possible. As for pathways using adenosine, a straightforward assumption is that these pathways start with adenosine kinase. However, a pathway bypassing this enzyme and using S-adenosylhomocysteine hydrolase instead was reported. So far, this route has not been analysed in detail. Using the concept of elementary flux modes, we investigate theoretically which salvage pathways exist in erythrocytes, which enzymes belong to each of these and what relative fluxes these enzymes carry. Here, we compute the net overall stoichiometry of ATP build-up from the recycled substrates and show that the network has considerable redundancy. For example, four different pathways of adenine salvage and 12 different pathways of adenosine salvage are obtained. They give different ATP/glucose yields, the highest being 3:10 for adenine salvage and 2:3 for adenosine salvage provided that adenosine is not used as an energy source. Implications for enzyme deficiencies are discussed. PMID- 16218957 TI - Molecular basis for substrate recognition and drug resistance from 1.1 to 1.6 angstroms resolution crystal structures of HIV-1 protease mutants with substrate analogs. AB - HIV-1 protease (PR) and two drug-resistant variants--PR with the V82A mutation (PR(V82A)) and PR with the I84V mutation (PR(I84V))--were studied using reduced peptide analogs of five natural cleavage sites (CA-p2, p2-NC, p6pol-PR, p1-p6 and NC-p1) to understand the structural and kinetic changes. The common drug resistant mutations V82A and I84V alter residues forming the substrate-binding site. Eight crystal structures were refined at resolutions of 1.10-1.60 A. Differences in the PR-analog interactions depended on the peptide sequence and were consistent with the relative inhibition. Analog p6(pol)-PR formed more hydrogen bonds of P2 Asn with PR and fewer van der Waals contacts at P1' Pro compared with those formed by CA-p2 or p2-NC in PR complexes. The P3 Gly in p1-p6 provided fewer van der Waals contacts and hydrogen bonds at P2-P3 and more water mediated interactions. PR(I84V) showed reduced van der Waals interactions with inhibitor compared with PR, which was consistent with kinetic data. The structures suggest that the binding affinity for mutants is modulated by the conformational flexibility of the substrate analogs. The complexes of PR(V82A) showed smaller shifts of the main chain atoms of Ala82 relative to PR, but more movement of the peptide analog, compared to complexes with clinical inhibitors. PR(V82A) was able to compensate for the loss of interaction with inhibitor caused by mutation, in agreement with kinetic data, but substrate analogs have more flexibility than the drugs to accommodate the structural changes caused by mutation. Hence, these structures help to explain how HIV can develop drug resistance while retaining the ability of PR to hydrolyze natural substrates. PMID- 16218959 TI - A human-specific TNF-responsive promoter for Goodpasture antigen-binding protein. AB - The Goodpasture antigen-binding protein, GPBP, is a serine/threonine kinase whose relative expression increases in autoimmune processes. Tumor necrosis factor (TNF) is a pro-inflammatory cytokine implicated in autoimmune pathogenesis. Here we show that COL4A3BP, the gene encoding GPBP, maps head-to-head with POLK, the gene encoding for DNA polymerase kappa (pol kappa), and shares with it a 140-bp promoter containing a Sp1 site, a TATA-like element, and a nuclear factor kappa B (NFkappaB)-like site. These three elements cooperate in the assembly of a bidirectional transcription complex containing abundant Sp1 and little NFkappaB that is more efficient in the POLK direction. Tumour necrosis factor cell induction is associated with Sp1 release, NFkappaB recruitment and assembly of a complex comparatively more efficient in the COL4A3BP direction. This is accomplished by competitive binding of Sp1 and NFkappaB to a DNA element encompassing a NFkappaB-like site that is pivotal for the 140-bp promoter to function. Consistently, a murine homologous DNA region, which contains the Sp1 site and the TATA-like element but is devoid of the NFkappaB-like site, does not show transcriptional activity in transient gene expression assays. Our findings identify a human-specific TNF-responsive transcriptional unit that locates GPBP in the signalling cascade of TNF and substantiates previous observations, which independently related TNF and GPBP with human autoimmunity. PMID- 16218960 TI - Abortive translation caused by peptidyl-tRNA drop-off at NGG codons in the early coding region of mRNA. AB - In Escherichia coli the codons CGG, AGG, UGG or GGG (NGG codons) but not GGN or GNG (where N is non-G) are associated with low expression of a reporter gene, if located at positions +2 to +5. Induction of a lacZ reporter gene with any one of the NGG codons at position +2 to +5 does not influence growth of a normal strain, but growth of a strain with a defective peptidyl-tRNA hydrolase (Pth) enzyme is inhibited. The same codons, if placed at position +7, did not give this effect. Other codons, such as CGU and AGA, at location +2 to +5, did not give any growth inhibition of either the wild-type or the mutant strain. The inhibitory effect on the pth mutant strain by NGG codons at location +5 was suppressed by overexpression of the Pth enzyme from a plasmid. However, the overexpression of cognate tRNAs for AGG or GGG did not rescue from the growth inhibition associated with these codons early in the induced model gene. The data suggest that the NGG codons trigger peptidyl-tRNA drop-off if located at early coding positions in mRNA, thereby strongly reducing gene expression. This does not happen if these codons are located further down in the mRNA at position +7, or later. PMID- 16218961 TI - Human mitochondrial complex I assembly is mediated by NDUFAF1. AB - Complex I (NADH:ubiquinone oxidoreductase) is the largest multiprotein enzyme of the oxidative phosphorylation system. Its assembly in human cells is poorly understood and no proteins assisting this process have yet been described. A good candidate is NDUFAF1, the human homologue of Neurospora crassa complex I chaperone CIA30. Here, we demonstrate that NDUFAF1 is a mitochondrial protein that is involved in the complex I assembly process. Modulating the intramitochondrial amount of NDUFAF1 by knocking down its expression using RNA interference leads to a reduced amount and activity of complex I. NDUFAF1 is associated to two complexes of 600 and 700 kDa in size of which the relative distribution is altered in two complex I deficient patients. Analysis of NDUFAF1 expression in a conditional complex I assembly system shows that the 700 kDa complex may represent a key step in the complex I assembly process. Based on these data, we propose that NDUFAF1 is an important protein for the assembly/stability of complex I. PMID- 16218962 TI - Type I antifreeze proteins expressed in snailfish skin are identical to their plasma counterparts. AB - Type I antifreeze proteins (AFPs) are usually small, Ala-rich alpha-helical polypeptides found in right-eyed flounders and certain species of sculpin. These proteins are divided into two distinct subclasses, liver type and skin type, which are encoded by separate gene families. Blood plasma from Atlantic (Liparis atlanticus) and dusky (Liparis gibbus) snailfish contain type I AFPs that are significantly larger than all previously described type I AFPs. In this study, full-length cDNA clones that encode snailfish type I AFPs expressed in skin tissues were generated using a combination of library screening and PCR-based methods. The skin clones, which lack both signal and pro-sequences, produce proteins that are identical to circulating plasma AFPs. Although all fish examined consistently express antifreeze mRNA in skin tissue, there is extreme individual variation in liver expression - an unusual phenomenon that has never been reported previously. Furthermore, genomic Southern blot analysis revealed that snailfish AFPs are products of multigene families that consist of up to 10 gene copies per genome. The 113-residue snailfish AFPs do not contain any obvious amino acid repeats or continuous hydrophobic face which typify the structure of most other type I AFPs. These structural differences might have implications for their ice-crystal binding properties. These results are the first to demonstrate a dual liver/skin role of identical type I AFP expression which may represent an evolutionary intermediate prior to divergence into distinct gene families. PMID- 16218963 TI - Si-face stereospecificity at C5 of coenzyme F420 for F420H2 oxidase from methanogenic Archaea as determined by mass spectrometry. AB - Coenzyme F420 is a 5-deazaflavin. Upon reduction, 1,5 dihydro-coenzyme F420 is formed with a prochiral centre at C5. All the coenzyme F420-dependent enzymes investigated to date have been shown to be Si-face stereospecific with respect to C5 of the deazaflavin, despite most F420-dependent enzymes being unrelated phylogenetically. In this study, we report that the recently discovered F420H2 oxidase from methanogenic Archaea is also Si-face stereospecific. The enzyme was found to catalyse the oxidation of (5S)-[5-2H1]F420H2 with O2 to [5-1H]F420 rather than to [5-2H]F420 as determined by MALDI-TOF MS. (5S)-[5-2H1]F420H2 was generated by stereospecific enzymatic reduction of F420 with (14a-2H2)-[14a-2H2] methylenetetrahydromethanopterin. PMID- 16218964 TI - Stem-loop oligonucleotides as tools for labelling double-stranded DNA. AB - We report on a sequence-specific double-stranded DNA labelling strategy in which a stem-loop triplex forming oligonucleotide (TFO) is able to encircle its DNA target. Ligation of this TFO to either a short hairpin oligonucleotide or a long double-stranded DNA fragment leads to the formation of a topological complex. This process requires the hybridization of both extremities of the TFO to each other on a few base pairs. The effects of different factors on the formation of these complexes have been investigated. Efficient complex formation was observed using both GT or TC TFOs. The stem-loop structure enhances the specificity of the complex. The topologically linked TFO remains associated with its target even under conditions that do not favour triple-helix formation. This approach is sufficiently sensitive for detection of a 20-bp target sequence at the subfemtomolar level. This study provides new insights into the mechanics and properties of stem-loop TFOs and their complexes with double-stranded DNA targets. It emphasizes the interest of such molecules in the development of new tools for the specific labelling of short DNA sequences. PMID- 16218965 TI - Purification and characterization of glutamate N-acetyltransferase involved in citrulline accumulation in wild watermelon. AB - Citrulline is an efficient hydroxyl radical scavenger that can accumulate at concentrations of up to 30 mm in the leaves of wild watermelon during drought in the presence of strong light; however, the mechanism of this accumulation remains unclear. In this study, we characterized wild watermelon glutamate N acetyltransferase (CLGAT) that catalyses the transacetylation reaction between acetylornithine and glutamate to form acetylglutamate and ornithine, thereby functioning in the first and fifth steps in citrulline biosynthesis. CLGAT enzyme purified 7000-fold from leaves was composed of two subunits with different N terminal amino acid sequences. Analysis of the corresponding cDNA revealed that these two subunits have molecular masses of 21.3 and 23.5 kDa and are derived from a single precursor polypeptide, suggesting that the CLGAT precursor is cleaved autocatalytically at the conserved ATML motif, as in other glutamate N acetyltransferases of microorganisms. A green fluorescence protein assay revealed that the first 26-amino acid sequence at the N-terminus of the precursor functions as a chloroplast transit peptide. The CLGAT exhibited thermostability up to 70 degrees C, suggesting an increase in enzyme activity under high leaf temperature conditions during drought/strong-light stresses. Moreover, CLGAT was not inhibited by citrulline or arginine at physiologically relevant high concentrations. These findings suggest that CLGAT can effectively participate in the biosynthesis of citrulline in wild watermelon leaves during drought/strong light stress. PMID- 16218966 TI - Proteasome involvement in the degradation of the G(q) family of Galpha subunits. AB - Metabolically unstable proteins are involved in a multitude of regulatory networks, including those that control cell signaling, the cell cycle and in many responses to physiological stress. In the present study, we have determined the stability and characterized the degradation process of some members of the G(q) class of heterotrimeric G proteins. Pulse-chase experiments in HEK293 cells indicated a rapid turnover of endogenously expressed Galpha(q) and overexpressed Galpha(q) and Galpha(16) subunits. Pretreatment with proteasome inhibitors attenuated the degradation of both G alpha subunits. In contrast, pretreatment of cells with inhibitors of lysosomal proteases and nonproteasomal cysteine proteases had very little effect on the stability of the proteins. Significantly, the turnover of these proteins is not affected by transient activation of their associated receptors. Fractionation studies showed that the rates of Galpha(q) and Galpha16 degradation are accelerated in the cytosol. In fact, we show that a mutant Galpha(q) which lacks its palmitoyl modification site, and which is localized almost entirely in the cytoplasm, has a marked increase in the rate of degradation. Taken together, these results suggest that the G(q) class proteins are degraded through the proteasome pathway and that cellular localization and/or other protein interactions determine their stability. PMID- 16218967 TI - A possible role of mitochondria in the apoptotic-like programmed nuclear death of Tetrahymena thermophila. AB - The ciliated protozoan Tetrahymena has a unique apoptosis-like process, which is called programmed nuclear death (PND). During conjugation, the new germinal micro and somatic macro-nuclei differentiate from a zygotic fertilized nucleus, whereas the old parental macronucleus degenerates, ensuring that only the new macronucleus is responsible for expression of the progeny genotype. As is the case with apoptosis, this process encompasses chromatin cleavage into high molecular mass DNA, oligonucleosomal DNA laddering, and complete degradation of the nuclear DNA, with the ultimate outcome of nuclear resorption. Caspase-8- and caspase-9-like activities are involved in the final resorption process of PND. In this report, we show evidence for mitochondrial association with PND. Mitochondria and the degenerating macronucleus were colocalized in autophagosome using two dyes for the detection of mitochondria. In addition, an endonuclease with similarities to mammalian endonuclease G was detected in the isolated mitochondria. When the macronuclei were incubated with isolated mitochondria in a cell-free system, DNA fragments of 150-400 bp were generated, but no DNA ladder appeared. Taking account of the present observations and the timing of autophagosome formation, we conclude that mitochondria might be involved in Tetrahymena PND, probably with the process of oligonucleosomal laddering. PMID- 16218968 TI - Microscopic observation of bacteria: review highlighting the use of environmental SEM. AB - Throughout the years, various methods have been adopted to investigate bacteria involved in root canal infection and apical periodontitis. This paper reviews the most commonly used microscopic techniques and discusses their possibilities, limitations and sample preparation. In particular, a recently developed variant of scanning electron microscope (SEM), referred to as environmental SEM (ESEM), is highlighted due to its potential impact across the diverse field of biomaterials research. The performance of this ESEM technique for bacterial observation of endodontic pathogens was illustrated by a practical approach. The paper concludes with a discussion on the possible use of ESEM for testing endodontic treatment modalities under environmental conditions in situ. PMID- 16218969 TI - The use of an intra-oral injection of ketorolac in the treatment of irreversible pulpitis. AB - AIM: To examine whether an intra-oral injection of a nonsteroidal anti inflammatory drug (ketorolac), in association with conventional local anaesthetic techniques, would improve the pulp extirpation rate in teeth with irreversible pulpitis. METHODOLOGY: A two group double-blind clinical trial was undertaken in the Dental Casualty Department of the University of Manchester School of Dentistry. Patients were randomly allocated to either the test or control group. The test group received an intra-oral injection of ketorolac (30 mg in 1 mL) in the buccal sulcus adjacent to the tooth being treated. After an interval of 15 min, they then received 2.2 mL of 2% lidocaine with 1 : 80 000 epinephrine by buccal infiltration in the maxilla or by inferior dental block in the mandible. The control group received an intra-oral injection of normal saline (1 mL) in the buccal sulcus adjacent to the tooth being treated, followed by the same local anaesthetic regime as the test group after the 15 min interval. Fifteen minutes after the local anaesthetic injections, pulp extirpation was attempted. All patients completed the short-form McGill pain questionnaire prior to treatment and completed identical questionnaires at 6 and 24 h after treatment. RESULTS: The study protocol set the number of patients to be treated at twenty. However, as the study progressed it became apparent that the intra-oral injection of ketorolac caused significant pain to four of the five patients who received it; therefore the study was terminated after ten patients had been treated. The results from the patients treated showed no significant difference in the pulp extirpation rate between the test and control groups. However, patients with higher pain scores at baseline were less likely to have the pulp completely extirpated, irrespective of whether they were in the test or control group. Pain scores for all patients decreased significantly from baseline to 24 h. CONCLUSION: An intra-oral injection of ketorolac did not improve the pulp extirpation rate in a small group of patients with irreversible pulpitis compared with a placebo. In addition, it was associated with such significant pain on injection that it cannot be recommended as a treatment in this situation. PMID- 16218971 TI - Physical and mechanical characterization and the influence of cyclic loading on the behaviour of nickel-titanium wires employed in the manufacture of rotary endodontic instruments. AB - AIM: To analyse the influence of cyclic loading on the mechanical behaviour of nickel-titanium (NiTi) wires employed in the manufacture of ProFile rotary endodontic instruments. METHODOLOGY: Nickel-titanium wires, 1.2 mm in diameter, taken from the production line of ProFile rotary endodontic instruments before the final machining step, were tensile-tested to rupture in the as-received condition and after 100 load-unload cycles in the superelastic plateau (4% elongation). The wires were characterized by X-ray energy-dispersive spectroscopy, X-ray diffraction and by differential scanning calorimetry and compared with new size 30, .06 taper ProFile instruments. The fracture surfaces of the wires were observed by scanning electron microscopy. RESULTS: The mechanical properties of the as-received wires, their chemical composition, the phases present and their transformation temperatures were consistent with their final application. Only small changes, which decreased after the first few cycles, took place in the mechanical properties of the cycled wires. The stress at maximum load and the plastic strain at breakage remained the same, while the critical stress for inducing the superelastic behaviour, which is related to the restoring force of the endodontic instruments, decreased by approximately 27%. CONCLUSIONS: The mechanical behaviour of the NiTi wires was modified slightly by cyclic tensile loading in the superelastic plateau. As the changes tended towards stabilization, the clinical use of rotary NiTi ProFile instruments does not compromise their superelastic properties until they fracture by fatigue or torsional overload, or are otherwise discarded. PMID- 16218972 TI - Defects in ProTaper S1 instruments after clinical use: fractographic examination. AB - AIM: To investigate the mode of failure of a brand of nickel-titanium instruments separated during clinical use, by detailed examination of the fracture surface. METHODOLOGY: A total of 122 ProTaper S1 instruments were discarded from an endodontic clinic at a stomatological school in China over a period of 17 months; 28 had fractured. These fractured instruments were ultrasonically cleaned, autoclaved and then examined under a scanning electron microscope. From the lateral view the fracture was classified into 'torsional' or 'flexural'. The specimens were then re-mounted and the presence of characteristics of shear failure and fatigue striations was recorded under high-power view of the fracture surface. The difference in the mean lengths of fractured segment between the shear and fatigue groups was compared using Student's t-test. RESULTS: Twenty seven separated instruments were available for analysis. Under low-power magnification, only two fell into the category of 'torsional' failure when examined laterally; the others appeared to be 'flexural'. Close examination of the fracture surface revealed the presence of fatigue striations in 18 specimens. Nine instruments (including the two putative 'torsional' failures above) fell into the shear fracture group, in which fatigue striations were absent or characteristics of shear failure of the material were found. The mean length of fractured segments resulting from fatigue failure (4.3+/-1.9 mm) was significantly greater than that for shear failure (2.5+/-0.8 mm) (P<0.001, two sample t-test). CONCLUSIONS: Examination of the fracture surface at high magnification is essential to reveal features that may indicate the possible origin of cracks and the mode of material failure. Macroscopic or lateral examination of separated instruments would fail to reveal the true mechanism of failure. Fatigue seems to be an important reason for the separation of rotary instruments during clinical use. PMID- 16218973 TI - Assessment of a gel-type chelating preparation containing 1-hydroxyethylidene-1, 1-bisphosphonate. AB - AIM: To test an aqueous gel containing 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) regarding its interactions with sodium hypochlorite, its calcium binding capacity, and its potential in preventing the formation of a smear layer when used in conjunction with rotary root canal preparation. METHODOLOGY: The experimental aqueous gel consisted of (w/v) 2% alginate, 3% aerosil, 10% Tween 80 and 18% HEBP. Interactions of gel components with hypochlorite were assessed using iodometric titration and monochromatic ultraviolet spectrometry. Two commercial paste-type chelators containing ethylenediaminetetraacetic acid (EDTA) and peroxide (RC-Prep and Glyde) served as controls. Calcium-binding capacities were measured in mixtures with a Ca2+ standard solution buffered at pH 10 using a calcium-selective measuring chain. Finally, root canals of 16 extracted single rooted premolars per group were instrumented using ProFile instruments dipped in the experimental gel, RC-Prep, or nothing. Additionally, canals were rinsed with 10 mL of a 1% NaOCl solution during/after preparation. Smear scores in instrumented teeth were monitored using scanning electron microscopy. RESULTS: None of the experimental gel components showed short-term interactions with hypochlorite, whilst EDTA, peroxide, RC-Prep and Glyde immediately reduced the hypochlorite in solution. The experimental gel chelated 30 mg Ca2+ g-1, compared with 16 mg Ca2+ g-1 and 11 mg Ca2+ g-1 chelated by RC-Prep and Glyde respectively. Smear scores obtained with the experimental gel were significantly (P<0.05) lower than with RC-Prep in coronal and middle root thirds, whilst no differences were observed in apical root thirds. CONCLUSIONS: Under the conditions of this study, an HEBP gel appeared advantageous over currently available products. PMID- 16218974 TI - Reduction of osteocalcin expression in aged human dental pulp. AB - AIM: To investigate the expression of osteocalcin mRNA in young and in aged human dental pulp tissue to determine the characteristics of osteocalcin expression. METHODOLOGY: Human dental pulp tissues of the third molars were obtained from healthy young (17-23 years) and aged (>50 years) subjects, and total RNA was extracted. Osteocalcin mRNA expression was determined by RT-PCR and by quantitative real-time RT-PCR (QRT-PCR). The threshold cycle (Ct) value, which reflects the amount of PCR, was calculated and the difference between the value in young and aged pulp was statistically analysed. RESULTS: Osteocalcin mRNA was detected in all samples of human dental pulp tissue homogenates by RT-PCR analysis. Osteocalcin mRNA was expressed in young adult dental pulp but was decreased in aged human dental pulp. QRT-PCR analysis also showed a reduced expression of osteocalcin mRNA in aged human pulp. Expression of osteocalcin in young human pulp was significantly higher (about sixfold) than in aged pulp (P<0.01, Mann-Whitney U-test). CONCLUSION: Reduction of osteocalcin expression may be associated with the loss of viability in human dental pulp tissue, and may be a characteristic of aged human dental pulps. PMID- 16218975 TI - Root morphology of mandibular premolars in 40 patients with Turner syndrome. AB - AIM: To analyse root morphology of mandibular premolars in 40 females with Turner syndrome. METHODOLOGY: All patients aged 18-50 years (mean+/-SD: 26.2+/-6.22) were clinically and cytogenetically diagnosed and divided into three groups according to karyotype: I-with 45,X (n=19); II-with 46,X,i(Xq) and other structural changes of this chromosome (n=8); III-with mos45,X/46,XX and other mosaic karyotype (n=13). The control group consisted of 30 healthy women aged 20 50 years (mean+/-SD: 31.52+/-5.21) who underwent dental treatment at the Department of Conservative Dentistry and Department of Periodontology and Oral Mucosa Diseases, Medical University of Gdansk, Poland. The root morphology of mandibular premolars was studied based on orthopantomogram X-ray images. The results were analysed using chi-square test with Yates's adjustment. RESULTS: Separate mesial and distal root canals and separate root apices were noted on orthopantomograms in patients with Turner syndrome. Two-rooted mandibular first premolars were observed in 34% of cases in group 45,X (group I), in 31% of cases in the group with structural aberrations of chromosome X (group II) and in 31% of cases with mosaic karyotypes (group III). Two-rooted mandibular second premolars were observed in 39% cases in group 45,X and in 31% of cases in the group with structural aberrations of chromosome X and in 35% of cases with mosaic karyotypes. This type of root morphology was not observed in the control group. No significant difference was found between the three groups of Turner syndrome. CONCLUSION: Highly significant differences in root morphology of mandibular premolars between patients with Turner syndrome and a population control group were found. PMID- 16218976 TI - Determining the apical terminus of root-end resected teeth using three modern apex locators: a comparative ex vivo study. AB - AIM: To assess ex vivo the accuracy of various electronic apex locators in locating the apical terminus of root-end resected teeth. METHODOLOGY: Ninety extracted human posterior teeth (182 root canals) were prepared to a minimum size of 40 and filled with gutta-percha and sealer. After resection of the apical 3 mm of the root, the root canal filling was removed using HERO rotary instruments. The size of the root canal at the apical terminus after removal of the filling ranged from size 50 to 90. The root canal length to the apical terminus was determined using 3 apex locators (Root ZX, Raypex4 and Apex Pointer). A new mounting model that utilized a micrometer was used to perform the measurements and to visually determine the actual position of the apical terminus. The frequency of locating the apical terminus and the corresponding 95% confidence interval (CI) were calculated. Additionally, the coefficient of repeatability of each apex locator and the limits of inter-operator agreement were determined. RESULTS: All apex locators showed an acceptable repeatability (0.02-0.03 mm coefficient of repeatability) and narrow limits of inter-operator agreement (+0.07 and -0.07 mm). The accuracy of determining the apical terminus within 1 mm in the root canal was as follows: Root ZX 90% (164/182 root-canals) [95%CI: 86 94%], Raypex4 74% (135/182 root-canals) [95%CI: 68-80%], and Apex Pointer 71% (129/182 root canals) [95%CI: 65-77%]. No over-instrumentation resulted when the Root ZX device was used. In contrast, using the Raypex4 or the Apex Pointer device resulted in over-instrumentation in 8 of 182 root canals (4%). CONCLUSIONS: Under the conditions of this study all three apex locators were able to detect the apical terminus of root-end resected teeth with an acceptable range. The Root ZX device was the most accurate without over-instrumentation of the root canals. PMID- 16218977 TI - The chemical constitution and biocompatibility of accelerated Portland cement for endodontic use. AB - AIM: To evaluate the biocompatibility of mineral trioxide aggregate and accelerated Portland cement and their eluants by assessing cell metabolic function and proliferation. METHODOLOGY: The chemical constitution of grey and white Portland cement, grey and white mineral trioxide aggregate (MTA) and accelerated Portland cement produced by excluding gypsum from the manufacturing process (Aalborg White) was determined using both energy dispersive analysis with X-ray and X-ray diffraction analysis. Biocompatibility of the materials was assessed using a direct test method where cell proliferation was measured quantitatively using Alamar Blue dye and an indirect test method where cells were grown on material elutions and cell proliferation was assessed using methyltetrazolium assay as recommended by the International standard guidelines, ISO 10993-Part 5 for in vitro testing. RESULTS: The chemical constitution of all the materials tested was similar. Indirect studies of the eluants showed an increase in cell activity after 24 h compared with the control in culture medium (P<0.05). Direct cell contact with the cements resulted in a fall in cell viability for all time points studied (P<0.001). CONCLUSIONS: Biocompatibility testing of the cement eluants showed the presence of no toxic leachables from the grey or white MTA, and that the addition of bismuth oxide to the accelerated Portland cement did not interfere with biocompatibility. The new accelerated Portland cement showed similar results. Cell growth was poor when seeded in direct contact with the test cements. However, the elution made up of calcium hydroxide produced during the hydration reaction was shown to induce cell proliferation. PMID- 16218978 TI - Neurological complications following extrusion of sodium hypochlorite solution during root canal treatment. AB - AIM: To report the presentation and management of two cases with neurological complications secondary to the extrusion of sodium hypochlorite solution into the facial soft tissues during root canal treatment. SUMMARY: The clinical features, with particular emphasis on nerve deficit following inadvertent extrusion of sodium hypochlorite, are discussed and its management highlighted. Early and aggressive treatment is advocated following such incidents in order to reduce potentially serious complications. KEY LEARNING POINTS: *Neurological sequelae can follow inadvertent hypochlorite extrusion. *Early recognition may avert a potentially more serious outcome. *Active hospital treatment including intravenous steroids and antibiotics is recommended. PMID- 16218980 TI - Staff shortages in the mental health workforce: the case of the disappearing approved social worker. AB - Approved social worker (ASW) numbers in England and Wales were compared on the basis of two national surveys conducted in 1992 and 2002. These data were supplemented by reports published by the Employers' Organisation in the intervening years. Although raw numbers suggested a modest absolute increase over this time, rates of ASW's per 100,000 population declined by over 50%. Possible explanations for this dramatic fall are explored. The authors conclude that specific and targeted action needs to be taken by the government and public sector employers to determine the numbers of mental health social workers needed in modernised community mental health services. PMID- 16218981 TI - Lay understandings of the effects of poverty: a Canadian perspective. AB - Although there is a large body of research dedicated to exploring public attributions for poverty, considerably less attention has been directed to public understandings about the effects of poverty. In this paper, we describe lay understandings of the effects of poverty and the factors that potentially influence these perceptions, using data from a telephone survey conducted in 2002 on a random sample (n=1671) of adults from eight neighbourhoods in two large Canadian cities (Edmonton and Toronto). These data were supplemented with interview data obtained from 153 people living in these same neighbourhoods. Multivariate linear and logistic regressions were used to determine the effects of basic demographic variables, exposure to poverty and attribution for poverty on three dependent variables relating to the effects of poverty: participation in community life, the relationship between poverty and health and challenges facing low-income people. Ninety-one per cent of survey respondents agreed that poverty is linked to health, while 68% agreed that low-income people are less likely to participate in community life. Affordable housing was deemed especially difficult to obtain by 96%, but other resources (obtaining healthy food, giving children a good start in life, and engaging in healthy behaviours) were also viewed as challenging by at least 70% of respondents. The regression models revealed that when controlling for demographics, exposure to poverty explained some of the variance in recognising the effects of poverty. Media exposure positively influenced recognition of the poverty-health link, and attending formal talks was strongly related to understanding challenges of poverty. Attributions for poverty accounted for slightly more of the variance in the dependent variables. Specifically, structural and sociocultural attributions predicted greater recognition of the effects of poverty, in particular the challenges of poverty, while individualistic attributions predicted less recognition. Older and female respondents were more likely to acknowledge the effects of poverty. Income was positively associated with recognition of the poverty-health link, negatively associated with understanding the challenges of low-income people, and unrelated to perceptions of the negative effect of poverty on participation in community life. PMID- 16218982 TI - Using pooled budgets to integrate health and welfare services: a comparison of experiments in England and Sweden. AB - The lack of collaboration between health, social and other welfare services is believed to impair efficiency and reduce effectiveness in addressing the complex problems of patients. Differences in funding streams, political accountabilities, organisational structures and professional cultures are all alleged to contribute to barriers between services. Drawing on their respective evaluations, this paper describes experiments in England and Sweden that use pooled budgets between services to improve interagency and interprofessional collaboration and presents evidence on their impact. Despite differences in the funding and organisation of health and welfare services in each country, some similar conclusions are reached. Among senior managers and politicians, budget pooling broadened their awareness of interdependencies with other agencies and professionals in promoting patients' welfare. However, these broadened perspectives were not immediately shared by professionals working at the front line, with whom patients had immediate contact. Moreover, neither experiment yielded unequivocal evidence of improved cost-effectiveness or of the benefits of budget pooling on the outcomes for service users. These experiments also raise questions about the equity and accountability of welfare services because in both countries only a limited range of services has been integrated under the umbrella of the pooled budgets. PMID- 16218983 TI - Who's there and who cares: age as an indicator of social support networks for caregivers among people living with motor neurone disease. AB - This paper explores the social support networks available to the informal carers of people living with motor neurone disease (MND). An ethnographic case study was undertaken using eco mapping, observation and conversational interviews to collect data from 18 primary carers of people living with MND. Interviews took place in participants' homes in metropolitan, regional and rural locations. Participants discussed the content of their support network and drew lines between individuals to indicate the type and strength of relationship. Changes to the network were depicted on eco maps during subsequent interviews. While health policy-makers assume that healthy social capital exists in Australian communities and that social cohesion will ensure active and available support networks in times of illness or disability, data from this exploratory study indicated that this was not consistently the case. Support networks varied in size and composition; however, age was identified as a discriminator of the availability and consistency of support. People in older age groups identified more diverse but consistent support systems while people in younger age groups reported more fluctuations in the strength of relationships and declines in support as caregiving became more demanding. Individual assessment of support networks at regular intervals in the caregiving trajectory is vital for all carers. However carers in younger age groups may need specific support to manage the psychological crises that occur and more access to paid care. Older carers may need consistent support to handle more of the instrumental aspects of care and assistance to mobilise their support networks. Community workers should be alert to the possible need for crisis intervention when tensions in relationships threaten carers' ability to provide effective care. PMID- 16218984 TI - Partnership working by default: district nurses and care home staff providing care for older people. AB - Older people residents in care homes that only offer residential care rely on primary health care services for medical and nursing needs. Research has investigated the demands that care homes staff and residents make on general practice, but not the involvement of other members of the primary health care team. This paper describes two consecutive studies completed in 2001 and 2003 that involved focus groups and survey methods of enquiry conducted in two settings: an England shire and inner London. The research questions that both studies had in common were (1) What is the contribution of district nursing and other primary care services to care homes that do not have on-site nursing provision? (2) What strategies promote participation and collaboration between residents, care home staff and NHS primary care nursing staff? and (3) What are the current obstacles and aids to effective partnership working and learning? A total of 74 community-based nurses and care home managers and staff took part in 10 focus groups, while 124 care home managers (73% of the 171 surveyed) and 113 district nurse team leaders (80% of the 142 surveyed) participated in the surveys. Findings from both studies demonstrated that nurses were the most frequent NHS professional visiting care homes. Although care home managers and district nurses believed that they had a good working relationship, they had differing expectations of what the nursing contribution should be and how personal and nursing care were defined. This influenced the range of services that older people had access to and the amount of training and support care home staff received from district nurses and the extent to which they were able to develop collaborative and reciprocal patterns of working. Findings indicate that there is a need for community-based nursing services to adopt a more strategic approach that ensures older people in care homes can access the services they are entitled to and receive equivalent health care to older people who live in their own homes. PMID- 16218985 TI - A national survey of adult placement schemes in England: recruitment and retention of adult placement carers. AB - Adult placement (AP) schemes and the carers they support have been, up to now, a largely invisible part of the social care workforce. Carers in particular fail to appear in workforce statistics. They provide a unique form of care to vulnerable people, under-pinned by values of extended family ('kinship') support. Little is known about the range of current activities of AP schemes and the carers they support. This study of AP schemes in England aimed to provide baseline data about the workforce, care provision, qualification levels, training and support issues, and recruitment and retention problems. Questionnaires were sent to all known schemes in England in both statutory and independent sectors. A final database of 130 schemes was compiled and of these, 124 contributed to the survey with complete or partial information. Variables relating to carers, and recruitment and retention problems experienced by schemes were analysed in detail. Age, gender and ethnic profiles of AP carers were described. Anecdotal evidence had suggested that the current regulatory system was leading to a loss of AP carers and/or an increasingly unregulated sector. Results from analyses supported this as the major reason given by schemes for the problems they had experienced in the last 12 months. Estimates of both the current level of registration and the numbers of carers cancelling their registration in the previous 12 months were made. Recently agreed regulatory changes aim to shift the burden of regulation from individual carers to AP schemes, and thereby ensure a consistent standard of assessment, training and support across England. Evidence from the survey provided support for such a move. PMID- 16218986 TI - Housing and dementia care - a scoping review of the literature. AB - This paper reports the findings of a scoping study designed to describe the evidence base with regard to housing provision for elderly people with dementia with the aim of identifying gaps in existing knowledge. This report from the scoping study findings covers studies of housing and accommodation in relation to dementia that have been published in the UK since the early 1980s, although we draw on limited aspects of overseas research to illuminate issues missing from the UK research agenda. The results reveal a significant number of research gaps in the UK context, most notably in relation to end-of-life care for people with dementia and the effectiveness of integrated and segregated facilities. UK policy regarding the development of extra-care housing also neglects the long-term future of people with dementia. A more robust evidence base will be required, combining a variety of methodological approaches, if UK dementia research is to contribute to future housing policy developments. PMID- 16218988 TI - Peptide chemical biology and drug design: a tribute issue to Editor-in-Chief Victor J. Hruby. PMID- 16218989 TI - Synthesis and biological evaluation of constrained analogues of the opioid peptide H-Tyr-D-Ala-Phe-Gly-NH2 using the 4-amino-2-benzazepin-3-one scaffold. AB - The synthesis of conformationally restricted dipeptidic moieties 4-amino-1,2,4,5 tetrahydro-2-benzazepin-3-one (Aba)-Gly ([(4S)-amino-3-oxo-1,2,4,5-tetrahydro-1H 2-benzazepin-2-yl]-acetic acid) and 8-hydroxy-4-amino-1,2,4,5-tetrahydro-2 benzazepin-3-one (Hba)-D-Ala ([(4S)-amino-8-hydroxy-3-oxo-1,2,4,5-tetrahydro benzo[c]azepin-2-yl]-propionic acid) was based on a synthetic strategy that uses an oxazolidinone as an N-acyliminium precursor. Introducing these Aba scaffolds into the N-terminal tetrapeptide of dermorphin (H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser NH2)-induced remarkable shifts in affinity and selectivity towards the opioid mu- and delta-receptors. This paper provides the synthesis and biological in vitro and in vivo evaluation of constricted analogues of the N-terminal tetrapeptide H Tyr-D-Ala-Phe-Gly-NH2, which is the minimal subunit of dermorphin needed for dermorphin-like opiate activity. PMID- 16218990 TI - A novel route to synthesize Freidinger lactams by micowave irradiation. AB - The incorporation of a Freidinger-like lactam structure into the backbone of peptides has been proven to be an useful strategy in the design of a variety of conformationally restricted targets. Several different strategies have been developed toward Freidinger lactams but no one resulted to be completely facile. Here, we report an efficient strategy that involves the iodo-derivatives in side chain of an appropriate amino acid used as electrophilic agent, and the standard solid phase peptide synthesis assisted by microwave irradiation. The methodology developed could be useful to perform Freidinger-like lactams with defined stereochemistry for routine use in solid phase peptide chemistry. PMID- 16218991 TI - Molecular mechanisms of constitutive activity: mutations at position 111 of the angiotensin AT1 receptor. AB - A possible molecular mechanism for the constitutive activity of mutants of the angiotensin type 1 receptor (AT1) at position 111 was suggested by molecular modeling. This involves a cascade of conformational changes in spatial positions of side chains along transmembrane helix (TM3) from L112 to Y113 to F117, which in turn, results in conformational changes in TM4 (residues I152 and M155) leading to the movement of TM4 as a whole. The mechanism is consistent with the available data of site-directed mutagenesis, as well as with correct predictions of constitutive activity of mutants L112F and L112C. It was also predicted that the double mutant N111G/L112A might possess basal constitutive activity comparable with that of the N111G mutant, whereas the double mutants N111G/Y113A, N111G/F117A, and N111G/I152A would have lower levels of basal activity. Experimental studies of the above double mutants showed significant constitutive activity of N111G/L112A and N111G/F117A. The basal activity of N111G/I152A was higher than expected, and that of N111G/Y113A was not determined due to poor expression of the mutant. The proposed mechanism of constitutive activity of the AT(1) receptor reveals a novel nonsimplistic view on the general problem of constitutive activity, and clearly demonstrates the inherent complexity of the process of G protein-coupled receptor (GPCR) activation. PMID- 16218992 TI - Manganese (IV) oxide-catalyzed electrophilic diamination of electron-deficient alkenes provides an easy synthesis of alpha,beta-diamino acid and ketone derivatives for peptidomimetic studies. AB - Manganese (IV) oxide was found to catalyze the diamination reaction of alpha,beta unsaturated esters and ketones with N,N-dichloro-p-toluenesulfonamide and acetonitrile as the halogen and nitrogen sources. The reaction is convenient to be conducted by simply mixing three reactants in the presence of manganese dioxide catalyst and 4 A molecular sieves, and provides an easy access to 1-p toluenesulfonyl-3-trichloromethyl-4,5-imidazoline derivatives, which are useful building blocks for peptidomimetic studies. PMID- 16218993 TI - Design of high affinity cyclic pentapeptide ligands for kappa-opioid receptors. AB - Using results from our previously reported cyclic opioid peptide series and reliable models for mu-, delta-, and kappa-opioid receptors (MOR, DOR, and KOR, respectively) and their complexes with peptide ligands, we have designed and synthesized a series of cyclic pentapeptides of structure Tyr-C[D-Cys-Phe-Phe-X] NH2, cyclized via disulfide, methylene, or ethylene dithioethers, and where X = D or L-Cys; or D- or L-penicillamine (Pen; beta,beta-dimethylcysteine). Determination of binding affinities to MOR, DOR, and KOR revealed that members of this series with X = D- or L-Cys display KOR affinities in the low nanomolar range, demonstrating that a 'DPDPE-like' tetrapeptide scaffold is suitable not only for DOR and MOR ligands, but also for KOR ligands. The cyclic pentapeptides reported here are not, however, selective for KOR, rather they display significant selectivity and high affinity for MOR. Indeed, peptide 8, Tyr-C[D-Cys Phe-Phe-Cys]-NH2-cyclized via a methylene dithioether, shows picomolar binding affinity for MOR ( = 16 pm) with more than 100-fold selectivity for MOR vs. DOR or KOR, and may be of interest as a high affinity, high selectivity MOR ligand. Nonetheless, the high affinity KOR peptides in this series represent excellent leads for the development of structurally related, selective KOR ligands designed to exploit structurally specific features of KOR, MOR, and DOR. PMID- 16218994 TI - Development of highly potent and selective dynorphin A analogues as new medicines. AB - Dynorphin A (Dyn A), a 17 amino acid peptide H-Tyr-Gly-Gly-Phe-Leu-Arg-Arg-Ile Arg-Pro-Lys-Leu-Lys-Trp-Asp-Asn-Gln-OH, is a potent opioid peptide which interacts preferentially with kappa-opioid receptors. Research in the development of selective and potent opioid peptide ligands for the kappa-receptor is important in mediating analgesia. Several cyclic disulphide bridge-containing peptide analogues of Dyn A, which were conformationally constrained in the putative message or address segment of the opioid ligand, were designed, synthesized and assayed. To further investigate the conformational and topographical requirements for the residues in positions 5 and 11 of these analogues, a systematic series of Dyn A(1-11)-NH2 cyclic analogues incorporating the sulphydryl-containing amino acids L- and D-Cys and L- and D-Pen in positions 5 and 11 were synthesized and assayed. Cyclic lactam peptide analogues were also synthesized and assayed. Several of these cyclic analogues, retained the same affinity and selectivity (vs. the mu- and delta-receptors) as the parent Dyn A(1 11)-NH2 peptide in the guinea-pig brain (GPB), but exhibited a much lower activity in the guinea-pig ileum (GPI), thus leading to centrally vs. peripherally selective peptides. Studies of the structure-activity relationship of Dyn A peptide provide new insights into the importance of each amino acid residue (and their configurations) in Dyn A analogues for high potency and good selectivity at kappa-opioid receptors. We report herein the progress towards the development of Dyn A peptide ligands, which can act as agonists or antagonists at cell surface receptors that modulate cell function and animal behaviour using various approaches to rational peptide ligand-based drug design. PMID- 16218995 TI - Tryptophan-containing peptide helices: interactions involving the indole side chain. AB - Two designed peptide sequences containing Trp residues at positions i and i + 5 (Boc-Leu-Trp-Val-Ala-Aib-Leu-Trp-Val-OMe, 1) as well as i and i + 6 (Boc-Leu-Trp Val-Aib-Ala-Aib-Leu-Trp-Val-OMe, 2) containing one and two centrally positioned Aib residues, respectively, for helix nucleation, have been shown to form stable helices in chloroform solutions. Structures derived from nuclear magnetic resonance (NMR) data reveal six and seven intramolecularly hydrogen-bonded NH groups in peptides 1 and 2, respectively. The helical conformation of octapeptide 1 has also been established in the solid state by X-ray diffraction. The crystal structure reveals an interesting packing motif in which helical columns are stabilized by side chain-backbone hydrogen bonding involving the indole Nepsilon1H of Trp(2) as donor, and an acceptor C=O group from Leu(6) of a neighboring molecule. Helical columns also associate laterally, and strong interactions are observed between the Trp(2) and Trp(7) residues on neighboring molecules. The edge-to-face aromatic interactions between the indoles suggest a potential C-H...pi interaction involving the Czeta3H of Trp(2). Concentration dependence of NMR chemical shifts provides evidence for peptide association in solution involving the Trp(2) Nepsilon1H protons, presumably in a manner similar to that observed in the crystal. PMID- 16218996 TI - Modified melanocortin tetrapeptide Ac-His-dPhe-Arg-Trp-NH at the arginine side chain with ureas and thioureas. AB - The Ac-His-dPhe-Arg-Trp-NH2 tetrapeptide is a nonselective melanocortin agonist and replacement of Arg in the tetrapeptide with acidic, basic or neutral amino acids results in reduced potency at the melanocortin receptor (MCR) isoforms (MC1R and MC3-5R). To determine the importance of the positive charge and the guanidine moiety for melanocortin activity, a series of urea- and thiourea substituted tetrapeptides were designed. Replacement of Arg with Lys or ornithine reduced agonist activity at the mouse mMC1 and mMC3-5 receptors, thus supporting the hypothesis that the guanidine moiety is important for receptor potency, particularly at the MC3-5 receptors. The Arg side chain-modified tetrapeptides examined in this study include substituted phenyl, naphthyl, and aliphatic urea and thiourea residues using a Lys side-chain template. These ligands elicit full agonist pharmacology at the mouse MCRs examined in this study. PMID- 16218997 TI - Regulation of the pituitary growth hormone-releasing hormone receptor in ageing male and female LOU rats: new insights into healthy ageing. AB - Ageing is characterised by a decrease of somatotroph functionality, involving growth hormone-releasing hormone receptor (GHRH-R). The present study was conducted in LOU/C/jall (LOU) rats, a strain described as a model of healthy ageing, which is characterised by a low adiposity and long life expectancy without developing severe pathologies. Effects of age and diet (chow versus self selection), on levels of anterior pituitary GHRH-R mRNA transcripts, were assessed in male and female LOU rats. The effect of age on pituitary GHRH-R functionality was examined in the anterior pituitary of both males and females fed chow diet. Moreover, serum insulin-like growth factor-I (IGF-I), T4 and leptin were measured because changes in their concentration could affect GHRH-R expression. In the pituitary of 18-month-old male and female LOU/C/jall rats fed standard chow, the level of 2.5-kb GHRH-R mRNA transcript, coding for functional GHRH-R, was significantly decreased. In 24- to 34-month-old males and females, it progressively returned to the level of younger animals, suggesting an enrichment of the group with survivors maintaining functional GHRH-R. In males and females repeatedly submitted to self-selection, this phenomenon was not observed. Studies with the GHRH-R agonist, Fluo-GHRH, revealed that 73% of 16-18-month-old male and female rats studied did not show an increase of fluorescence density characteristic of receptor-mediated internalisation upon incubation at 37 degrees C. In the other 27%, the increase of fluorescence was identical to that observed in pituitaries of young rats, suggesting the presence of an optimal level of functional GHRH-R. Serum levels of leptin, free T4 and total IGF-I decreased more drastically in ageing males and in rats fed a self-selection diet. A positive correlation was demonstrated between leptin and IGF-I levels in ageing males and females fed standard chow and ageing females submitted to a self-selection regimen. In conclusion, healthy ageing in LOU rats fed chow diet appears to be associated with a maintenance of functional pituitary GHRH-R levels found in younger rats but not necessarily with those of serum leptin, T4 and IGF-I. PMID- 16218998 TI - Growth hormone-releasing peptide-6 increases insulin-like growth factor-I mRNA levels and activates Akt in RCA-6 cells as a model of neuropeptide Y neurones. AB - Chronic systemic administration of growth hormone (GH)-releasing peptide-6 (GHRP 6), an agonist for the ghrelin receptor, to normal adult rats increases insulin like growth factor (IGF)-I mRNA and phosphorylated Akt (pAkt) levels in various brain regions, including the hypothalamus. Because neuropeptide Y (NPY) neurones of the arcuate nucleus express receptors for ghrelin, we investigated whether these neurones increase their IGF-I and p-Akt levels in response to this agonist. In control rats, immunoreactive pAkt was practically undetectable; however, GHRP 6 increased p-Akt immunoreactivity in the arcuate nucleus, with a subset of neurones also being immunoreactive for NPY. Immunoreactivity for IGF-I was detected in NPY neurones in both experimental groups. To determine if activation of this intracellular pathway is involved in modulation of NPY synthesis RCA-6 cells, an embryonic rat hypothalamic neuronal cell line that expresses NPY was used. We found that GHRP-6 stimulates NPY and IGF-I mRNA synthesis and activates Akt in this cell line. Furthermore, inhibition of Akt activation by LY294002 treatment did not inhibit GHRP-6 induction of NPY or IGF-I synthesis. These results suggest that some of the effects of GHRP-6 may involve stimulation of local IGF-I production and Akt activation in NPY neurones in the arcuate nucleus. However, GHRP-6 stimulation of NPY production does not involve this second messenger pathway. PMID- 16218999 TI - Hypothalamic energy balance gene responses in the Sprague-Dawley rat to supplementation of high-energy diet with liquid ensure and subsequent transfer to chow. AB - Energy dense, high fat, high sugar, foods and beverages in our diet are a major contributor to the escalating global obesity problem. Here, we examine the physiological and neuroendocrine effects of feeding rats a solid high-energy (HE) diet with or without a liquid supplement (Ensure) and the consequence of subsequently transferring animals back to chow (C). Outbred Sprague-Dawley rats were fed C until 49-56 days of age, and then transferred a HE diet for 3 weeks before allocation to one of two weight-matched groups. Over the next 10 weeks, one group remained on HE diet, whereas the other had access to the liquid diet, chocolate Ensure (EN), in addition to HE diet (HE + EN). Half the rats from each group were then killed, and the remainder were returned to C for 3 weeks. Supplementation of the HE diet with EN accelerated weight gain and increased daily energy intake, adipose tissue mass, and circulating leptin levels. Transferring animals back to C caused a decrease in bodyweight in the HE + EN group, whereas HE animals were weight stable. Both groups also exhibited voluntary hypophagia, although the magnitude and duration of this response was greater in HE + EN animals. The only effect of Ensure on the hypothalamic genes studied was on tyrosine kinase B expression in the ventromedial hypothalamic nucleus (VMH), which was increased in rats given the supplement. Withdrawal of the obesogenic diets decreased gene expression for cocaine-and-amphetamine regulated transcript (CART) and dynorphin (DYN) in the arcuate nucleus (ARC), and DYN and brain-derived neurotrophic factor (BDNF) in the VMH, whereas neuropeptide Y (NPY) gene expression in the ARC was increased. These changes were independent of previous dietary history. EN supplementation generates distinct physiological responses, yet has a minimal effect on hypothalamic neuropeptide or receptor gene expression, possibly due to the development of leptin resistance. Withdrawal of obesogenic diets induces changes in the gene expression consistent with NPY, CART and BDNF attempting to oppose weight gain on either HE or HE + EN. PMID- 16219001 TI - Formalin attenuates the stress-induced increase in plasma epinephrine levels. AB - Subcutaneous (s.c.) injection of formalin into rats is frequently used as a painful stressor that produces a three-phase nociceptive response. We have shown previously that s.c. administered formalin (0.2 ml of 4% solution per 100 g body weight) unexpectedly attenuated the increase of plasma epinephrine levels in rats exposed to exteroceptive stressors (handling, immobilisation). To clarify the mechanism(s) responsible for this phenomenon, the effect of formalin applications on epinephrine plasma levels was investigated in various experimental conditions. Subcutaneous application of formalin combined with exposures of animals to an interoceptive stressor, insulin-induced hypoglycaemia, significantly attenuated the stress-induced increase in plasma epinephrine levels, whereas plasma norepinephrine levels remained highly elevated. Moreover, administration of formalin to unstressed animals also manifested signs of an attenuated epinephrine secretion. Interestingly, intraperitoneal administration of formalin did not reduce the elevated levels of plasma epinephrine. We suggest that formalin attenuates epinephrine secretion from the adrenal medulla most probably via irritation of s.c. somatosensory receptors. We hypothesise that the irritation of the primary sensory afferents fibres might reduce the activity of the sympathetic preganglionic neurones innervating adrenal medullary chromaffin cells. Further investigations are required to establish whether the observed reduction of epinephrine secretion from the adrenal medulla is controlled by either spinal or supraspinal neuronal circuits. PMID- 16219000 TI - Hypothalamic phosphatidylinositol 3-kinase-phosphodiesterase 3B-cyclic AMP pathway of leptin signalling is impaired following chronic central leptin infusion. AB - Leptin signalling in the hypothalamus is critical for the maintenance of normal body weight. Although hyperleptinaemia in obese people suggests a state of leptin resistance, and diet-induced obesity in rodents is associated with central leptin resistance, the underlying mechanisms remain unclear. Recent evidence suggests that, in addition to the signal transducer and activator of the transcription-3 (STAT3) pathway, leptin action is critical for energy homeostasis through an insulin-like signalling pathway involving an increase in phosphatidylinositol 3 kinase (PI3K) and phosphodiesterase 3B (PDE3B) activities and reduction in cyclic AMP (cAMP) levels in the hypothalamus. Here, we show that chronic central leptin (160 ng/h) infusion, which resulted in the development of resistance to the satiety action of leptin, impaired the PI3K-PDE3B-cAMP pathway of leptin signalling in the hypothalamus in that PI3K and PDE3B activities were increased and cAMP levels were decreased in the hypothalamus on day 2 of leptin infusion but remained unchanged on day 16. Additionally, induction of tyrosyl phosphorylation of insulin receptor substrate-1 observed on day 2 was not evident on day 16 of leptin infusion. By contrast, signalling through the STAT3-pathway remained activated in the hypothalamus throughout 16 days of leptin infusion. These findings show a differential response in PI3K-PDE3B-cAMP (impaired) and STAT3 (up-regulated) pathways to chronic central leptin infusion, and suggest a selective resistance in the PI3K-PDE3B-cAMP pathway of leptin signalling following a chronic increase in hypothalamic leptin tone attained by central infusion of this peptide hormone. PMID- 16219002 TI - Skeleton photoperiods alter delayed-type hypersensitivity responses and reproductive function of Siberian hamsters (Phodopus sungorus). AB - Photoperiod (day length) can modulate immune function. Whether these photoperiodic effects on immune function are mediated directly by a circadian photoperiodic time measurement system or indirectly by nonspecific (e.g. stressful) effects of light is unknown. To discriminate between these two possibilities, Siberian hamsters (Phodopus sungorus) were housed in either long or short photoperiods (LD 16 : 8 h or LD 8 : 16 h) or in 'skeleton' long or short photoperiods (LD 1 : 14 h: LD 1 : 8 h or LD 1 : 6 h: LD 1 : 16 h). In the skeleton photoperiods, both long- and short-day animals received 2 h of light per day. After 10 weeks in their respective photoperiods, hamsters were tested for an antigen specific immune response using a delayed type hypersensitivity (DTH) model. Reproductive and endocrine responses of hamsters in each of the skeleton photoperiods were equivalent to those in standard long or short days, respectively. Hamsters in skeleton short days and LD 8 : 16 increased DTH responses compared to hamsters in both long-day groups. DTH responses were equivalent in both long-day groups. These results suggest that the influences of day length on immune function potentially are due to circadian photoperiodic time measurement systems. PMID- 16219003 TI - Genetic differences in hypothalamic-pituitary-adrenal axis activity and food restriction-induced hyperactivity in three inbred strains of rats. AB - We used three inbred rat strains known for significant differences in the activity and reactivity of their hypothalamic-pituitary-adrenal (HPA) axis to stress [Fischer 344 (F344), Brown Norway (BN) and Lewis (Lew) rats] to search for a strain difference in the paradoxical increase in running activity induced by food restriction and to explore the role of the HPA axis in this behaviour. Rats were randomly assigned to either an ad lib sedentary group (AL), a control wheel activity group (ACT), a food restriction-induced hyperactivity group (FR-ACT) group (1.5 h/day ad lib food, 22.5 h/day ad lib wheel access) or a pair-fed group (FR). The BN and Lew rats reached the 25% body weight-loss criterion of FR-ACT (strain effect: F(2,132) = 45.58, P < 10-6) faster than the F344 strain due to higher food restriction-induced running activity (strain effect: F(2,65) = 17.43, P = 0.00001). FR and FR-ACT decreased thymus weight (marker of integrated HPA axis activation) in all strains. In Lew and BN strains, FR-ACT induced a further decrement on thymus weight compared to their FR group. Prefeeding corticosterone levels (15.00 h) increased during the study in BN and Lew FR-ACT rats, but not in F344. Total wheel turns were correlated to both final adipose weight (r = -0.49, P = 0.002) and thymus weight decrement (r = 0.59, P = 0.0001), emphasizing the relationship between fat mass and HPA axis activation in excessive running activity. Increased running in conditions of food restriction and HPA axis activation may be linked at the level of the central nervous system. However, the involvement of corticotrophin-releasing hormone, agouti-related peptide or cocaine- and amphetamine-regulated transcript in behavioural disturbances of FR ACT rats was excluded (in situ hybridization). We propose that corticosterone may be the link between initial low levels of fat mass and/or rate of fat mass loss (peripheral energy stores) and increased wheel activity, favouring fueling through lipolysis and proteolysis and reinforcing the self starvation via reward mechanisms, thus establishing a deleterious vicious cycle. PMID- 16219004 TI - Smad proteins are targets of transforming growth factor beta1 in immortalised gonadotrophin-releasing hormone releasing neurones. AB - Transforming growth factor beta (TGFbeta) is one of the growth factors involved in the neuroendocrine control of the gonadotrophin-releasing hormone (GnRH) neurones. It is produced and released by the astrocytes surrounding GnRH neurones and directly controls their secretory activity. TGFbeta signalling is based on a complex of two receptors that transduces the signal through peculiar intracellular substrates, the Smad proteins, which, upon activation, move into the nucleus, and modify the transcription of TGFbeta responsive genes. The present study aimed to verify whether TGFbeta1 is able to regulate the Smad pathway in GT1-1 cells (i.e. an immortalised neuronal cell line releasing GnRH). We show that: (i) GT1-1 cells express Smad 2, 3, 4, and 7; (ii) TGFbeta1 enhances the phosphorylation of Smad 2 and 3 at short times of exposure (15-30 min); (iii) TGFbeta1 induces the synthesis of the inhibitory Smad 7 at longer times (60-120 240 min); (iv) the conditioned medium of type 1 astrocytes enhances the phosphorylation of Smad 2 and 3 in GT1-1 cells and a TGFbeta1 neutralising antibody counteracts this effect. The results indicate that Smads are targets of TGFbeta1 and that astrocytes are able to modulate Smads proteins in GT1-1 cells through the release of TGFbeta1. Taken together, the data provide new evidence that glial cells are important regulators of the GnRH neuronal activity. PMID- 16219006 TI - Molecular mechanisms of lipid-induced insulin resistance in muscle, liver and vasculature. AB - Increased body fat content correlates with insulin resistance and is a key feature of type 2 diabetes. Excessive intake of fat results in deposition of lipids not only in fat tissue but also in skeletal muscle and liver. Subsequently, both plasma and intracellular concentrations of free fatty acids and their metabolites rise and activate signal transduction pathways, which will induce inflammation and impair insulin signalling. Furthermore, elevated circulating lipids impair endothelial function and fibrinolysis, which contributes to the development of vascular disease. Thus, therapeutic strategies aiming at reduction of (intracellular) lipid availability in skeletal muscle and liver and pharmacological modulation of the signalling pathways activated by increased lipid stores represent promising targets for future treatment of insulin resistance and prevention of its complications. This review focuses on the effects of increased lipid availability on the regulation of glucose metabolism in skeletal muscle and liver as well as on vascular function. PMID- 16219005 TI - Chronic intermittent cold stress sensitises the hypothalamic-pituitary-adrenal response to a novel acute stress by enhancing noradrenergic influence in the rat paraventricular nucleus. AB - Chronic intermittent cold stress sensitises activation of the hypothalamic pituitary-adrenal (HPA) axis by novel acute stress. We have shown that enhanced noradrenergic function in limbic forebrain contributes to HPA sensitisation. In the present study, we investigated whether chronic intermittent cold also induced changes in noradrenergic function in the paraventricular nucleus (PVN), the primary mediator of the HPA stress response. Rats were exposed to chronic intermittent cold (7 days, 6 h per day, 4 degrees C). On the day after final cold exposure, there were no differences in baseline plasma ACTH, but the peak ACTH response to 30 min of acute immobilisation stress was greater in cold-stressed rats compared to controls. Bilateral microinjection of the alpha(1)-adrenergic receptor antagonist benoxathian into the PVN reduced acute stress-induced adrenocorticotrophic hormone (ACTH) levels by approximately 25% in controls. Furthermore, in cold-stressed rats, all of the sensitisation of the ACTH response was blocked by benoxathian, to a level comparable to benoxathian-treated controls. In a second study using microdialysis to measure norepinephrine release in the PVN, there were no differences in either baseline or acute stress-induced increases in norepinephrine release in the PVN of cold-stressed rats compared to controls. Thus, in a third study, we tested potential alterations in postsynaptic alpha(1)-receptor sensitivity after chronic cold stress. Dose-dependent activation of ACTH secretion by microinjection of the alpha(1)-adrenergic receptor agonist, phenylephrine, into the PVN was significantly enhanced in cold stressed rats compared to controls. Thus, the sensitised HPA response to acute stress after chronic intermittent cold exposure is at least partly attributable to an enhanced response to alpha1-adrenergic receptor activation in the PVN. Chronic stress-induced plasticity in the acute stress response may be important for stress adaptation, but may also contribute to pathophysiological conditions associated with stress. Thus, understanding the neural mechanisms underlying such adaptations may help us understand the aetiology of such disorders, and contribute to the future development of more effective treatment or prevention strategies. PMID- 16219007 TI - Combination therapy using metformin or thiazolidinediones and insulin in the treatment of diabetes mellitus. AB - The biguanide, metformin, sensitizes the liver to the effect of insulin, suppressing hepatic glucose output. Thiazolidinediones such as rosiglitazone and pioglitazone enhance insulin-mediated glucose disposal, leading to reduced plasma insulin concentrations. These classes of drugs may also have varying beneficial effects on features of insulin resistance such as lipid levels, blood pressure and body weight. Metformin in combination with insulin has been shown to significantly improve blood glucose levels while lowering total daily insulin dose and body weight. The thiazolidinediones in combination with insulin have also been effective in lowering blood glucose levels and total daily insulin dose. Triple combination therapy using insulin, metformin and a thiazolidinedione improves glycaemic control to a greater degree than dual therapy using insulin and metformin or insulin and a thiazolidinedione. There is insufficient evidence to recommend the use of metformin or thiazolidinediones in type 1 diabetic patients. Although these agents are largely well tolerated, some subjects experience significant gastrointestinal problems while using metformin. Metformin is associated with a low risk of lactic acidosis, but should not be used in patients with elevated serum creatinine or those being treated for congestive heart failure. The thiazolidinediones are associated with an increase in body weight, although this can be avoided with careful lifestyle management. Thiazolidinediones may also lead to oedema and are associated with a low incidence of hepatocellular injury. Thiazolidinediones are contraindicated in patients with underlying heart disease who are at risk of congestive heart failure and in patients who have abnormal hepatic function. The desired blood glucose-lowering effect and adverse event profiles of these agents should be considered when recommending these agents to diabetic patients. The potential for metformin or the thiazolidinediones to impact long-term cardiovascular outcomes remains under investigation. PMID- 16219008 TI - Insulin resistance, diabetes and cardiovascular risk: approaches to treatment. AB - The prevalence of diabetes is increasing worldwide. Insulin resistance and diabetes mellitus are major predictors of cardiovascular ischaemic disease. Other risk factors for cardiovascular death including hypertension, dyslipidaemia, smoking and visceral obesity are especially lethal in diabetics. C-reactive protein, plasminogen activator inhibitor-1, matrix metalloproteinases and other emerging risk factors and their roles are continually being researched and discovered. Treatment of this syndrome must be aimed at lifestyle modification, glycaemic control and management of concomitant risk factors. Diet and exercise play a vital role in the treatment of diabetes and the metabolic syndrome. Weight reduction and increased physical activity will improve insulin resistance, hyperglycaemia, hypertension and dyslipidaemia. Hypertension management has been shown to be especially important in diabetics to prevent cardiovascular events. Likewise, multiple clinical trials show that reduction of cholesterol is even more vital in diabetics than the general population for risk reduction of coronary disease. There is a great deal of evidence that tight control of glycaemia is essential to treatment of this condition. There are a variety of available pharmacological agents available including metformin, insulin secretagogues, alpha-glucosidase inhibitors, thiazolidinediones and insulin. The mechanisms and side effects of these medications are discussed. As macrovascular disease is the major cause of morbidity and mortality, an early, aggressive, multi-factorial approach to treatment of the metabolic syndrome and diabetes is vital to prevent adverse cardiac outcomes. PMID- 16219009 TI - Metformin revisited: re-evaluation of its properties and role in the pharmacopoeia of modern antidiabetic agents. AB - BACKGROUND: The usefulness of metformin as an oral antidiabetic agent is widely accepted. However, several other classes of oral antidiabetic agents have been recently introduced, raising the need to evaluate the role of metformin as initial therapy and in combination with these newer drugs for treatment of type 2 diabetes mellitus (DM). METHODS: Synthesis of information was preceded by a comprehensive review of the English language literature using Medline. We also reviewed bibliographies of relevant articles. The studies most pertinent to the mechanism of action, efficacy, toxicity and administration of metformin were selected for citation in this review. RESULTS: Metformin acts by increasing tissue sensitivity to insulin, principally in the liver. Beneficial properties of metformin include weight reduction, favourable effects on the lipid profile and the fibrinolytic pathway, and improvement of ovarian function in some insulin resistant women. It does not cause hyperinsulinaemia or hypoglycaemia. Metformin is effective as monotherapy and, in combination with both insulin secretagogues and thiazolidinediones (TZDs), may obviate the need for insulin treatment. Several fixed-dose combination pills containing metformin and other agents are available. A protocol for the initiation of therapy with contemporary oral agents for type 2 DM is presented, with emphasis on the continuing central role of metformin. CONCLUSIONS: Metformin remains a safe and effective agent for the therapy of patients with type 2 DM. It is useful as monotherapy or in combination regimens with the newer insulin secretagogues, TZDs or insulin. It is still in most circumstances the agent of choice for initial therapy of the typical obese patient with type 2 DM and mild to moderate hyperglycaemia. PMID- 16219010 TI - Pharmacologic modifications of hormones to improve their therapeutic potential for diabetes management. AB - Rapid-acting genetically engineered insulin analogues emerging in the last 10 years are now established as more effective prandial insulins than traditional short-acting human insulin. The development of analogues for use as basal insulin, however, has been much slower. Methods of pro-tracting the time-action curve of injected insulin include complexing with proteins, insulin crystal formation, shifting the iso-electric point of the amino acid sequence or attaching a fatty-acid side chain to the molecule. The latter two methods have been more successful in producing physiologic insulin profiles when compared with the former methods. The principle of acylation has also been applied to prolong the action of other hormones, such as glucagon-like peptide 1 (GLP-1), as the native peptide has a very short half-life. Preliminary results with this compound and other GLP-1 analogues show promise in treating patients with type 2 diabetes. In summary, the development of new insulin and other hormone preparations by the manipulation of native peptide structure has recently improved our antidiabetic armamentarium, and further research will continue this fruitful approach. PMID- 16219011 TI - Treating insulin resistance in type 2 diabetes with metformin and thiazolidinediones. AB - Insulin resistance underlies the pathogenesis of hyperglycaemia and cardiovascular disease in most people with type 2 diabetes. Metformin and thiazolidinediones (pioglitazone and rosiglitazone) counter insulin resistance by different cellular mechanisms and with complementary effects, making them suited for use in combination. Metformin exerts a stronger suppression of hepatic glucose output, while thiazolidinediones produce a greater increase in peripheral glucose uptake, enabling metformin-thiazolidinedione combinations to improve glycaemic control in type 2 diabetes with additive efficacy. Basal insulin concentrations are not raised by metformin or thiazolidinediones, so there is minimal risk of hypoglycaemia, and metformin can reduce the weight gain associated with thiazolidinediones. There are overlapping effects of metformin and thiazolidinediones against a range of athero-thrombotic factors and markers. These include decreased plasminogen activator inhibitor-1, reduced platelet aggregation, reductions of several vascular adhesion molecules, and reduced markers of low-grade inflammation such as C-reactive protein. Additionally, thiazolidinediones increase adiponectin and slightly reduce blood pressure. Both metformin and thiazolidinediones can improve components of the lipid profile: thiazolidinediones consistently reduce free fatty acid concentrations and decrease the proportion of small dense low-density-lipoprotein, and pioglitazone also decreases triglycerides. During co-administration, metformin and thiazolidinediones do not interfere with each other's pharmacokinetics, and lower doses of the two agents together can achieve efficacy with fewer side effects. Metformin-thiazolidinedione combinations require attention to the precautions for both agents, especially renal, cardiac and hepatic status. Thus, metformin and thiazolidinediones can be used in combination to address the hyperglycaemia and vascular risk in type 2 diabetes. PMID- 16219012 TI - Improved glycaemic control with dipeptidyl peptidase-4 inhibition in patients with type 2 diabetes: vildagliptin (LAF237) dose response. AB - OBJECTIVE: A novel treatment option for diabetic patients is the enhancement of incretin hormone activity by inhibition of the enzyme dipeptidyl peptidase-4 (DPP 4). This study was designed to establish a dose of the DPP-4-inhibitor vildagliptin (LAF237) that was effective in reducing HbA1c levels and was safe and well tolerated in patients with type 2 diabetes. PATIENTS AND METHODS: The study of 279 patients with type 2 diabetes consisted of a 4-week run-in phase where patients received placebo and a 12-week active treatment phase where they received one of the following dosages of vildagliptin: 25 mg twice daily, 25, 50 or 100 mg once daily (qd), or placebo. RESULTS: There was a statistically significant reduction in HbA1c levels in the vildagliptin 50 mg qd (p=0.003) and 100 mg qd groups (p=0.004) compared with the placebo group. The mean 4-h postprandial glucose level was significantly reduced from placebo in the vildagliptin 50 mg qd group (p = 0.012) and mean 4-h postprandial insulin was significantly increased from baseline vs. placebo in the vildagliptin 100 mg qd group (p=0.022). The assessment of beta-cell function (HOMA-B) was significantly increased in the vildagliptin 100 mg qd treatment group (p=0.007). The incidence of adverse events was similar in all treatment groups including placebo. CONCLUSIONS: Vildagliptin, at 50 and 100 mg qd, was effective in reducing HbA1c levels compared with placebo in patients with type 2 diabetes. Vildagliptin at dosages up to 100 mg qd appeared safe and well tolerated. PMID- 16219013 TI - X-PERT: weight reduction with orlistat in obese subjects receiving a mildly or moderately reduced-energy diet: early response to treatment predicts weight maintenance. AB - AIM: To determine the effect of two different levels of energy deficit on weight loss in obese patients treated with orlistat. METHODS: Patients (n=430) were randomized in a 1-year, multicentre, open-label, parallel group study conducted at 23 hospital centres and university medical departments worldwide. Obese outpatients (body mass index 30--43 kg/m(2)) aged 18--70 years with a body weight of >or=90 kg and a waist circumference of >or=88 cm (women) or >or=102 cm (men) were treated with orlistat 120 mg three times daily plus a diet that provided an energy deficit of either 500 or 1,000 kcal/day for 1 year. Orlistat treatment was discontinued in patients who did not achieve >or=5% weight loss after assessment at 3 and 6 months. The primary outcome measure was change in body weight from baseline at week 52. RESULTS: Reported mean difference in energy intake between the two groups (500-1,000 kcal/day deficit) at weeks 24 and 52 was actually 111 and 95 kcal/day respectively. Of the 430 patients involved in the study, 295 achieved >or=5% weight loss at both 3 and 6 months. In this population, at week 52, weight loss from baseline was similar for patients randomized to either the 500 or the 1,000 kcal/day deficit diet (-11.4 kg vs. -11.8 kg, respectively; p=0.778). After 12 months of treatment with orlistat, 84% (n=118/141) and 85% (n=131/154) of patients in the 500 and 1,000 kcal/day deficit groups, respectively, achieved >or=5% weight loss, and 50% (n=70/141) and 53% (n=82/154) of patients, respectively, achieved >or=10% weight loss. Patients in both the diet treatment groups showed similar significant improvements in blood pressure, lipid levels and waist circumference at week 52. CONCLUSIONS: Treatment with orlistat was associated with a clinically beneficial weight loss, irrespective of the prescribed dietary energy restriction (-500 or -1000 kcal/day). Patients who achieved >or=5% weight loss at 3 months achieved long-term, clinically beneficial weight loss with orlistat plus either diet. Therefore, identifying patients who lose at least 5% weight after 3 months and who maintain this weight loss up to 6 months is a valuable treatment algorithm to select patients who will benefit most from orlistat treatment in combination with diet. PMID- 16219014 TI - Fermented mushroom milk-supplemented dietary fibre prevents the onset of obesity and hypertriglyceridaemia in Otsuka Long-Evans Tokushima fatty rats. AB - AIM: Fermented milk product containing edible mushroom water extracts (mushroom yogurt; MY) has been reported to have glycaemic control and triglyceride-lowering effects in streptozotocin (STZ)-induced diabetic rats and Zucker diabetic fatty (ZDF) rats. Here, we investigated how MY-supplemented dietary fibre (10 and 20%, v/w) influences the onset of obesity and hypertriglyceridaemia in Otsuka Long Evans Tokushima fatty (OLETF) rats. METHODS: The OLETF rats were fed a powdered chow diet supplemented with MY at the levels of 10 (v/w) and 20% for 6 weeks from 10 weeks of age, but the OLETF control rats were not supplemented. Their weight, fat distribution and lipid profile have been determined. RESULTS: The body weights in MY-fed rats were reduced compared with the control rats. The perirenal fat was decreased in both MY groups, but the visceral and epididymal fats reduced only in the MY 20% group. The concentrations of serum triglyceride and non esterified fatty acid in MY-fed rats were decreased in a dose-dependent manner. However, the levels of other serum lipid profiles [total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol] were comparable among all rats. CONCLUSION: Anti-obesity and triglyceride lowering by MY-supplemented dietary fibre in OLETF rats might have resulted from the synergistic effect of components in the fermented mushroom-milk product. PMID- 16219015 TI - Percentile distribution of the waist circumference among Mexican pre-adolescents of a primary school in Mexico City. AB - AIM: The aim of this study is to describe the percentile distribution of the waist circumference in Mexican pre-pubertal children living in an urban, middle income area. METHODS: Children, aged 6-10, from a primary school, located in a middle-income area in Mexico City were included (n=833). Anthropometric measurements were made in the case of all children by using standardized procedures. RESULTS: The prevalence of obesity and overweight was greater among boys (p<0.001). Of them, 30 were obese (7.2%) and 47 were overweight (11.4%). Girls had a prevalence of obesity of 2.8% and of overweight of 8.8%. Boys had greater waist circumference than girls (p<0.001). This observation was independent from differences in age or body mass index (BMI). The distribution of the waist circumference was shifted towards higher values among boys. The values corresponding to the 85th percentile ranged, in boys aged 6-10, between 62.9 and 85.9 cm. By contrast, for girls, those values were 66.8 and 72.08 cm, respectively. A receiver-operator curve was constructed in order to identify the waist circumference most likely to be associated with a BMI of > 85th percentile. We found that age- and sex-specific cut-off points are required for the detection of central adiposity in children aged 6-10. CONCLUSIONS: The percentile distribution of the waist circumference in Mexican pre-pubertal children living in urban, middle-income areas is described in this study. Moreover, included are the cut-off points most likely to be associated with increased adiposity. This information can be used for defining abdominal obesity in paediatric populations. PMID- 16219016 TI - The DIACOM study (effect of DosIng frequency of oral Antidiabetic agents on the COMpliance and biochemical control of type 2 diabetes). AB - AIM: Sulphonylureas are widely used in the management of type 2 diabetes. The effectiveness of treatment with oral antidiabetic drugs depends largely on patient compliance. The objective of the DIACOM (effect of DosIng frequency of oral Antidiabetic agents on the COMpliance and biochemical control of type 2 diabetes) study was to compare the compliance of patients treated with once-daily (od) or twice-daily (bid) sulphonylureas. METHODS: One hundred and five patients, previously treated with glibenclamide, were randomized to receive gliclazide in modified-release formulation (MR) once daily or glibenclamide twice daily for 16 weeks, using an electronic monitoring system (MEMS). RESULTS: A significant difference in compliance was observed between the two groups. The overall compliance was 93.5+/-14.0% in the once-daily gliclazide MR group and 87.2+/ 21.1% in the twice-daily glibenclamide group (p<0.05), and the correct number of doses was taken on 86.3+/-15.4 and 66.9+/-29.0% of treatment days respectively (p<0.0001). The percentage of missed doses was 9.3+/-12.5% in the once-daily group and 17.5+/-18.0% in the twice-daily group (p<0.01). The percentage of doses taken in the correct time window and correct inter-dose interval was higher in the once-daily group, as was therapeutic coverage. Patients in the gliclazide MR group also achieved significantly better glycaemic control [fasting plasma glucose and glycated haemoglobin (HbA(1c))] than those treated with glibenclamide (p<0.0001). CONCLUSIONS: The study demonstrates that patient compliance with once daily gliclazide MR is significantly better than with twice-daily glibenclamide. Consistently better efficacy was observed for short-term (fasting glucose) and long-term glycaemic control (HbA(1c)) in the once-daily group. These results demonstrate the possible therapeutic advantages of once-daily agents over twice daily agents in the treatment of type 2 diabetes. PMID- 16219017 TI - Plasma IL-6 concentration is inversely related to insulin sensitivity, and acute phase proteins associate with glucose and lipid metabolism in healthy subjects. AB - AIM: It has been shown that atherosclerosis is an inflammatory disease. Recent data suggest that inflammation precedes type 2 diabetes. Hence, we wanted to study the interrelationship between IL-6, insulin sensitivity, lipids and numerous acute-phase proteins. METHODS: Twenty-one healthy individuals [16 males/5 females, age 27.9+/-1.8 years, body mass index (BMI) 24.1+/-0.8 kg/m(2)] participated in the study. Each patient went through a 4-h hyperinsulinaemic (40 mU/m(2)/min) euglycaemic clamp and 4-h saline infusion. Blood samples were taken before and at the end of the infusions. RESULTS: Plasma interleukin (IL)-6 concentration correlated inversely with insulin sensitivity (M-value) (r=-0.49, p<0.05). Moreover, the plasma levels of IL-6 associated with c-peptide (r=0.49, p<0.05), fat% (r=0.43, p<0.05) and diastolic blood pressure (r=0.46, p<0.05). alpha-1-acid glycoprotein was related to HbA1(c) (r=0.47, p<0.05), insulin (r=0.55, p<0.01), diastolic blood pressure (r=0.58, p<0.01), systolic blood pressure (r=0.58, p<0.01) and triglycerides (r=0.58, p<0.01). Haptoglobin was correlated with insulin (r=0.46, p<0.05), total cholesterol (r=0.61, p<0.01), BMI (r=0.58, p<0.01), fat% (r=0.63, p<0.01) and lipid oxidation during clamp (r=0.43, p<0.05). Diastolic blood pressure decreased during the clamp (from 78.3+/-1.9 to 72.1+/-2.0 mmHg, p=0.001). Insulin infusion did not affect the serum levels of most acute-phase proteins. CONCLUSIONS: Our study suggests that low grade inflammation, as reflected by IL-6, A1GP and haptoglobin contributes to the regulation of insulin sensitivity, lipid metabolism and blood pressure in normal human physiology. PMID- 16219018 TI - Relationship between pedometer-registered activity, aerobic capacity and self reported activity and fitness in patients with type 2 diabetes. AB - AIM: The aim of this study is to investigate the relationship between pedometer registered activity, aerobic capacity (VO(2 max)) and self-reported activity and fitness in patients with type 2 diabetes before and after a 12-week exercise programme. METHODS: Twenty-nine men with type 2 diabetes (age=57.4 (7.8) years, BMI=31.7 (2.8) kg/m(2)) underwent exercise testing, registered pedometer activity and reported their physical fitness and activity in a questionnaire. Participants were randomly allocated to an exercise (EX) group (n=15) or a control (CO) group (n=14). Participants in EX group were offered supervised exercise twice a week for 12 weeks. At the end of the study, participants again underwent exercise testing, fasting blood tests and registration of pedometer activity. RESULTS: At baseline, pedometer activity correlated with VO(2 max) (r=0.43, p=0.02) and with perceived physical fitness (r=0.48, p=0.02). After, but not before, intervention, pedometer activity and VO(2 max) additionally correlated with perceived everyday activity (r=0.62, p<0.01 and r=0.49, p=0.03, respectively). Both EX and CO groups tended to increase pedometer activity. In EX group, weight decreased by 2.7% (p=0.01), VO(2 max) increased 10.6% (p=0.03) and HbA1c decreased by 5.2% from baseline (p=0.02). In EX group, an increase in pedometer activity correlated with a fall in HbA1c (r=0.84, p<0.01) and in diastolic blood pressure (r=0.77, p=0.025). CONCLUSIONS: Pedometer activity correlates with VO(2 max) in type-2 diabetic patients. Our exercise programme was well tolerated; it produced favourable effects on body weight, aerobic capacity and metabolic control. The use of pedometer may lead to more realistic assessment of perceived everyday physical activity. PMID- 16219019 TI - Role of skeletal muscle-fibre type in regulation of glucose metabolism in middle aged subjects with impaired glucose tolerance during a long-term exercise and dietary intervention. AB - AIM: The aim of this study was to investigate the role of skeletal muscle fibre type in the regulation of glucose metabolism in middle-aged obese subjects with impaired glucose tolerance (IGT) during a 2-year exercise and dietary intervention. METHODS: Muscle biopsies (musculus vastus lateralis) were taken from 22 subjects belonging to the intervention group of the Finnish Diabetes Prevention Study [1]. According to their myosin heavy chain (MHC) profile at the baseline, the subjects were divided into two groups: IGT(slow) (n=10) with a high proportion of MHC I isoforms and IGT(fast) (n=12) with a high proportion of MHC II isoforms in the vastus lateralis muscle. The intervention consisted of dietary counselling, strength and power training and/or aerobic exercise. The amount of exercise was the same in both groups; the exercise frequency was 5.1+/-2.7 h/week in the IGT(slow) and 5.1+/-2.8 h/week in the IGT(fast) group. RESULTS: Fasting glucose (p<0.05), 2-h glucose (p<0.05), fasting insulin (p<0.05), haemoglobin A1c (HbA(1c)) (p<0.01) and insulin resistance (p<0.05) [homeostasis model assessment for insulin resistance (HOMA-IR)] decreased in the IGT(fast) group, whereas only the 2-h glucose and HbA(1c) concentrations decreased in the IGT(slow) group. The amount of the glycogen synthase kinase-3-alphabeta (GSK-3-alphabeta) decreased in the IGT(fast) group (p<0.05). Exercise training increased the lactate dehydrogenase (LDH) (p<0.01), LDH-1 (p<0.05) and citrate synthase (CS) (p<0.05) activities in the vastus lateralis muscle in the IGT(slow) group, but only the CS activity (p<0.05) in the IGT(fast) group. CONCLUSIONS: The glucose metabolism improved both in the IGT(slow) and IGT(fast) group during the 2-year exercise and dietary intervention. The change was more prominent in the IGT(fast) group than in the IGT(slow) group, associated with the decrease of the GSK-alphabeta protein expression in skeletal muscle. The exercise training improved both glycolytic and oxidative capacity in the vastus lateralis muscle. The glycolytic capacity improved in the IGT(slow) group and the oxidative capacity in both groups. PMID- 16219020 TI - Effect of three treatment schedules of recombinant methionyl human leptin on body weight in obese adults: a randomized, placebo-controlled trial. AB - AIM: The aim of this study was to evaluate the effect on body weight and safety of subcutaneously administered recombinant leptin in obese adults and to evaluate whether the timing of recombinant leptin administration influences efficacy. METHODS: A randomized, double-blind, placebo-controlled, multicentre study was designed, comprising of a 3-week dietary lead-in followed by a 12-week leptin or placebo treatment period. A total of 284 overweight and obese (body mass index 27 37.0 kg/m(2)) predominantly white (98%) women (66%) and men (34%) with a mean (+/ s.d.) 46.8+/-10.4 years of age were randomized into three treatment groups with three matching placebo groups. Recombinant leptin was administered by subcutaneous injection [10 mg/morning, 10 mg/evening or 20 mg/day (10 mg twice daily)]. Patients were counselled at baseline to reduce dietary intake by 2,100 kJ/day (500 kcal/day), and dietary advice was reinforced every 2-4 weeks. RESULTS: No statistically significant change in body weight occurred with recombinant leptin treatment compared with placebo treatment in any treatment group. No clinically significant adverse effects were observed with the exception of an increase in injection-site reactions in patients treated with recombinant leptin (83%) vs. placebo (36%). CONCLUSIONS: Administration of recombinant leptin to an overweight and obese population, in addition to a mildly energy-restricted diet, was not efficacious in terms of weight loss at the doses and schedules studied. The hypothesis that nocturnal administration of recombinant leptin might have a specific effect on weight loss was not supported. PMID- 16219021 TI - Re-evaluation of exercise intensity for overweight Japanese men by ventilatory threshold. AB - AIM: To re-evaluate exercise intensity for overweight Japanese men by ventilatory threshold (VT). METHODS: Cross sectional clinical intervention study. SUBJECTS AND MATERIALS: One hundred and ten overweight Japanese men aged 32-59 years were recruited. The average body mass index was 28.5+/-2.5 kg/m(2). Aerobic exercise level was evaluated by measuring VT. Fifty per cent heart rate (HR) reserve by Karvonen's formula was calculated and compared with HR at VT. Fat distribution was evaluated by visceral fat and subcutaneous fat areas measured with computed tomography scanning at umbilical levels. Anthropometric parameters such as height, body weight and body fat percentage were also measured. RESULTS: There was significant correlation between HR at VT and 50% HR reserve by Karvonen's formula (r=0.642, p<0.01). However, HR at VT was significantly lower than 50% HR reserve by Karvonen's formula (HR at VT 103.6+/-10.6 beat/min vs. 50% HR reserve 125.0+/-7.1 beat/min, p<0.01) and HR at VT was corresponded to 28.5+/-8.3% HR reserve by Karovonen's formula. CONCLUSION: The present study indicated that exercise intensity of 30% HR reserve should be recommended for overweight Japanese men. PMID- 16219025 TI - Transcriptional profiling of spinal cord injury-induced central neuropathic pain. AB - Central neuropathic pain (CNP) is an important problem following spinal cord injury (SCI), because it severely affects the quality of life of SCI patients. As in the patient population, the majority of rats develop significant allodynia (CNP rats) after moderate SCI. However, about 10% of SCI rats do not develop allodynia, or develop significantly less allodynia than CNP rats (non-CNP rats). To identify transcriptional changes underlying CNP development after SCI, we used Affymetrix DNA microarrays and RNAs extracted from the spinal cords of CNP and non-CNP rats. DNA microarry analysis showed significantly increased expression of a number of genes associated with inflammation and astrocytic activation in the spinal cords of rats that developed CNP. For example, mRNA levels of glial fibrilary acidic protein (GFAP) and Aquaporin 4 (AQP4) significantly increased in CNP rats. We also found that GFAP, S100beta and AQP4 protein elevation persisted for at least 9 months throughout contused spinal cords, consistent with the chronic nature of CNP. Thus, we hypothesize that CNP development results, in part, from dysfunctional, chronically "over-activated" astrocytes. Although, it has been shown that activated astrocytes are associated with peripheral neuropathic pain, this has not previously been demonstrated in CNP after SCI. PMID- 16219024 TI - The mitochondrial complex I inhibitor rotenone triggers a cerebral tauopathy. AB - Reduced activity of the mitochondrial respiratory chain--particularly complex I- may be implicated in the etiology of both Parkinson's disease and progressive supranuclear palsy, although these neurodegenerative diseases differ substantially as to their distinctive pattern of neuronal cell loss and the predominance of cerebral alpha-synuclein or tau protein pathology. To determine experimentally whether chronic generalized complex I inhibition has an effect on the distribution of alpha-synuclein or tau, we infused rats systemically with the plant-derived isoflavonoid rotenone. Rotenone-treated rats with a pronounced metabolic impairment had reduced locomotor activity, dystonic limb posture and postural instability. They lost neurons in the substantia nigra and in the striatum. Spherical deposits of alpha-synuclein were observed in a few cells, but cells with abnormal cytoplasmic accumulations of tau immunoreactivity were significantly more numerous in the striatum of severely lesioned rats. Abnormally high levels of tau immunoreactivity were found in the cytoplasm of neurons, oligodendrocytes and astrocytes. Ultrastructurally, tau-immunoreactive material consisted of straight 15-nm filaments decorated by antibodies against phosphorylated tau. Many tau+ cell bodies also stained positive for thioflavin S, nitrotyrosine and ubiquitin. Some cells with abnormal tau immunoreactivity contained activated caspase 3. Our data suggest that chronic respiratory chain dysfunction might trigger a form of neurodegeneration in which accumulation of hyperphosphorylated tau protein predominates over deposits of alpha-synuclein. PMID- 16219026 TI - Carboxypeptidase E is required for normal synaptic transmission from photoreceptors to the inner retina. AB - Defects in the gene encoding carboxypeptidase E (CPE) in either mouse or human lead to multiple endocrine disorders, including obesity and diabetes. Recent studies on Cpe-/- mice indicated neurological deficits in these animals. As a model system to study the potential role of CPE in neurophysiology, we carried out electroretinography (ERG) and retinal morphological studies on Cpe-/- and Cpe fat/fat mutant mice. Normal retinal morphology was observed by light microscopy in both Cpe-/- and Cpe(fat/fat) mice. However, with increasing age, abnormal retinal function was revealed by ERG. Both Cpe-/- and Cpe fat/fat animals had progressively reduced ERG response sensitivity, decreased b-wave amplitude and delayed implicit time with age, while maintaining a normal a-wave amplitude. Immunohistochemical staining showed specific localization of CPE in photoreceptor synaptic terminals in wild-type (WT) mice, but in both Cpe-/- and Cpe fat/fat mice, CPE was absent in this layer. Bipolar cell morphology and distribution were normal in these mutant mice. Electron microscopy of retinas from Cpe fat/fat mice revealed significantly reduced spherule size, but normal synaptic ribbons and synaptic vesicle density, implicating a reduction in total number of vesicles per synapse in the photoreceptors of these animals. These results suggest that CPE is required for normal-sized photoreceptor synaptic terminal and normal signal transmission to the inner retina. PMID- 16219027 TI - Inhibition of thrombin-induced microglial activation and NADPH oxidase by minocycline protects dopaminergic neurons in the substantia nigra in vivo. AB - The present study shows that activation of microglial NADPH oxidase and production of reactive oxygen species (ROS) is associated with thrombin-induced degeneration of nigral dopaminergic neurons in vivo. Seven days after thrombin injection in the rat substantia nigra (SN), tyrosine hydroxylase immunocytochemistry showed a significant loss of nigral dopaminergic neurons. This cell death was accompanied by localization of terminal deoxynucleotidyl transferase-mediated fluorecein UTP nick-end labelling (TUNEL) staining within dopaminergic neurons. This neurotoxicity was antagonized by the semisynthetic tetracycline derivative, minocycline, and the observed neuroprotective effects were associated with the ability of minocycline to suppress NADPH oxidase-derived ROS production and pro-inflammatory cytokine expression, including interleukin 1beta and inducible nitric oxide synthase, from activated microglia. These results suggest that microglial NADPH oxidase may be a viable target for neuroprotection against oxidative damage. PMID- 16219028 TI - Cocaine-induced CREB phosphorylation in nucleus accumbens of cocaine-sensitized rats is enabled by enhanced activation of extracellular signal-related kinase, but not protein kinase A. AB - Repeated cocaine administration to rats outside their home cages sensitizes the behavioral effects of the drug, and enhances induction of the immediate early gene product Fos in nucleus accumbens. We hypothesized that the same treatment regimen would also enhance cocaine-induced activation of intracellular signaling kinases that phosphorylate cyclic AMP-regulated element-binding protein (CREB), an important mediator of c-fos transcription. Phosphorylation levels of extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK), calcium/calmodulin kinases (CaMKs) II and IV, and CREB were used to assess endogenous functional activity of these signaling molecules in rats behaviorally sensitized outside their home cages. Protein kinase A (PKA)-specific phosphorylation of Ser845 in the alpha-amino-3-hydroxy-5-methylisoxazole-4 propionate (AMPA) receptor subunit GluR1 was used to assess endogenous functional activity of PKA. Using western blots and immunohistochemistry, we detected cocaine-induced CREB phosphorylation after repeated cocaine administration, but not after repeated saline administration. Using western blots and MAPK activity assays, we found that cocaine-induced phosphorylation and activation of ERK, but not of CaMKs II or IV or GluR1, was augmented in nucleus accumbens of cocaine sensitized rats. Unilateral infusions of the MAPK kinase inhibitor U0126 into nucleus accumbens attenuated cocaine-induced ERK and CREB phosphorylation in cocaine-sensitized rats. In contrast, unilateral infusions of the PKA inhibitor Rp-isomer of adenosine-3',5'-cyclicmonophosphorothioate (Rp-cAMPs) did not affect cocaine-induced CREB phosphorylation. Therefore, enhanced activation of ERK, but not PKA, enables and mediates cocaine-induced CREB phosphorylation in nucleus accumbens of rats that are sensitized by repeated cocaine administration outside their home cages. PMID- 16219030 TI - Neuronal protection by sirtuins in Alzheimer's disease. AB - Silent information regulator 2, a member of NAD+-dependent histone deacetylase in yeast, and its homologs in mice and humans, participate in numerous important cell functions, including cell protection and cell cycle regulation. The sirtuin family members are highly conserved evolutionarily, and are predicted to have a role in cell survival. The science of sirtuins is an emerging field and is expected to contribute significantly to the role of sirtuins in healthy aging in humans. The role of sirtuins in neuronal protection has been studied in lower organisms, such as yeast, worms, flies and rodents. Both yeast Sir2 and mammalian sirtuin proteins are up-regulated under calorie-restricted and resveratrol treatments. Increased sirtuin expression protects cells from various insults. Caloric restriction and antioxidant treatments have shown useful effects in mouse models of aging and Alzheimer's disease (AD) and in limited human AD clinical trials. The role sirtuins may play in modifying and protecting neurons in patients with neurodegenerative diseases is still unknown. However, a recent report of Huntington's disease revealed that Sirtuin protects neurons in a Huntington's disease mouse model, suggesting that sirtuins may protect neurons in patients with neurodegenerative diseases, such as AD. In this review, we discuss the possible mechanisms of sirtuins involved in neuronal protection and the potential therapeutic value of sirtuins in healthy aging and AD. PMID- 16219029 TI - In vivo interaction between serotonin and galanin receptors types 1 and 2 in the dorsal raphe: implication for limbic seizures. AB - The neuropeptide galanin suppresses seizure activity in the hippocampus by inhibiting glutamatergic neurotransmission. Galanin may also modulate limbic seizures through interaction with other neurotransmitters in neuronal populations that project to the hippocampus. We examined the role of galanin receptors types 1 and 2 in the dorsal raphe (DR) in the regulation of serotonergic transmission and limbic seizures. Infusion of a mixed agonist of galanin receptors types 1 and 2 [galanin (1-29)] into the DR augmented the severity of limbic seizures in both rats and wild-type mice and concurrently reduced serotonin concentration in the DR and hippocampus as measured by immunofluorescence or HPLC. In contrast, injection of the galanin receptor type 2 agonist galanin (2-11) mitigated the severity of seizures in both species and increased serotonin concentration in both areas. Injection of both galanin fragments into the DR of galanin receptor type 1 knockout mice exerted anticonvulsant effects. Both the proconvulsant activity of galanin (1-29) and seizure suppression by galanin (2-11) were abolished in serotonin-depleted animals. Our data indicate that, in the DR, galanin receptors types 1 and 2 modulate serotonergic transmission in a negative and a positive fashion, respectively, and that these effects translate into either facilitation or inhibition of limbic seizures. PMID- 16219032 TI - Progression of age-associated cognitive impairment correlates with quantitative and qualitative loss of TrkA receptor protein in nucleus basalis and cortex. AB - A direct correlation between disease progression and reduced expression of TrkA receptor in cholinergic neurons has been documented in neurocognitive pathologies including Alzheimer's disease. We investigated whether reduced expression of TrkA protein might also correlate with the level of cognitive impairment in age associated cognitive impairment. Quantitative and qualitative measurements of TrkA protein levels in the cortex and nucleus basalis of aged rats that had been well-characterized behaviorally as 'unimpaired', 'mildly impaired' or 'fully impaired' demonstrated significant changes in TrkA expression. In the mildly impaired cognitive state phenotypic silencing of TrkA was detected in neurons expressing TrkA at high density but before cholinergic atrophy or loss of TrkA+ neurons was detected. In the fully impaired cognitive state a significant loss in TrkA+ cholinergic neurons together with a more significant phenotypic silencing of TrkA expression then took place. These data suggest that TrkA+ cholinergic cells are associated with cognition, TrkA could be a biomarker of the cognitive state and phenotypic loss of TrkA precedes neuronal loss and probably sensitizes cells to death. We speculate that neurotrophic deficits may be a shared mechanism for cognitive decline in aging and Alzheimer's disease. PMID- 16219031 TI - Sustained pharmacological blockade of NK1 substance P receptors causes functional desensitization of dorsal raphe 5-HT 1A autoreceptors in mice. AB - Antagonists at NK1 substance P receptors have demonstrated similar antidepressant properties in both animal paradigms and in human as selective serotonin reuptake inhibitors (SSRIs) that induce desensitization of 5-HT 1A autoreceptors within the dorsal raphe nucleus (DRN). We investigated whether this receptor adaptation also occurs upon NK1 receptor blockade. C57B/L6J mice were treated for 21 days with the selective NK1 receptor antagonist GR 205171 (10 mg/kg daily) through subcutaneously implanted osmotic mini pumps, and DRN 5-HT 1A autoreceptor functioning was assessed using various approaches. Recording of DRN serotonergic neurons in brainstem slices showed that GR 205171 treatment reduced (by approximately 1.5 fold) the potency of the 5-HT 1A receptor agonist, ipsapirone, to inhibit cell firing. In parallel, the 5-HT 1A autoreceptor-mediated [35S]GTP gamma-S binding induced by 5-carboxamidotryptamine onto the DRN in brainstem sections was significantly decreased in GR 205171-treated mice. In vivo microdialysis showed that the cortical 5-HT overflow caused by acute injection of the SSRI paroxetine (1 mg/kg) was twice as high in GR 205171-treated as in vehicle-treated controls. In the DRN, basal 5-HT outflow was significantly enhanced by GR 205171 treatment. These data supported the hypothesis that chronic NK1 receptor blockade induces a functional desensitization of 5-HT 1A autoreceptors similar to that observed with SSRIs. PMID- 16219033 TI - Human spastin has multiple microtubule-related functions. AB - Hereditary spastic paraplegias (HSPs) are neurodegenerative diseases caused by mutations in more than 20 genes, which lead to progressive spasticity and weakness of the lower limbs. The most frequently mutated gene causing autosomal dominant HSP is SPG4, which encodes spastin, a protein that belongs to the family of ATPases associated with various cellular activities (AAAs). A number of studies have suggested that spastin regulates microtubule dynamics. We have studied the ATPase activity of recombinant human spastin and examined the effect of taxol-stabilized microtubules on this activity. We used spastin translated from the second ATG and provide evidence that this is the physiologically relevant form. We showed that microtubules enhance the ATPase activity of the protein, a property also described for katanin, an AAA of the same spastin subgroup. Furthermore, we demonstrated that human spastin has a microtubule destabilizing activity and can bundle microtubules in vitro, providing new insights into the molecular pathogenesis of HSP. PMID- 16219034 TI - Apparent presence of Ser133-phosphorylated cyclic AMP response element binding protein (pCREB) in brain mitochondria is due to cross-reactivity of pCREB antibodies with pyruvate dehydrogenase. AB - Cyclic AMP response element binding protein (CREB) is a constitutive transcription factor that activates transcription following stimulus-dependent phosphorylation at Ser133, implicated in synaptic plasticity and neuronal survival pathways. The prevailing view that CREB is exclusively nuclear has been questioned by several studies, and, for example, mitochondrial localization has been reported. Using subcellular fractionation of rat brain cortex coupled with western immunoblotting with Ser133-phospho-CREB (pCREB) antibodies, we found a robust pCREB immunoreactivity (IR) in mitochondria-enriched fractions. The pCREB antibodies also stained the mitochondria, in addition to nuclei, of glial cells in primary cortical cultures. However, two CREB antibodies against different epitopes and gel shift assay detected the CREB protein mainly in the nuclear fraction. The two-dimensional electrophoretic mobility of mitochondrial pCREB IR differed markedly from the nuclear CREB/pCREB IR, indicating that the pCREB antibody cross-reacts with another mitochondrial protein. Immunoprecipitation of the mitochondrial pCREB IR produced three bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis, which were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry as E2, E1 alpha subunit, and E1 beta-subunit of pyruvate dehydrogenase complex. The cross reacting epitope was identified as phospho-Ser300 of the alpha-subunit. In conclusion, this study confirms the presence of pCREB-like IR in brain mitochondria that, after careful scrutiny, turned out to be pyruvate dehydrogenase rather than authentic CREB. PMID- 16219035 TI - Characterization of glycoconjugate antigens in mouse embryonic neural precursor cells. AB - Neuronal and glial cells organizing the central nervous system (CNS) are generated from common neural precursor cells (NPCs) during neural development. However, the expression of cell-surface glycoconjugates that are crucial for determining the properties and biological function of these cells at different stages of development has not been clearly defined. In this study, we investigated the expression of several stage-specific glycoconjugate antigens, including several b-series gangliosides GD3, 9-O-acetyl GD3, GT1b and GQ1b, stage specific embryonic antigen-1 (SSEA-1) and HNK-1, in mouse embryonic NPCs employing immunocytochemistry and flow cytometry. In addition, several of these antigens were positively identified by chemical means for the first time. We further showed that the SSEA-1 immunoreactivity was contributed by both glycoprotein and glycolipid antigens, and that of HNK-1 was contributed only by glycoproteins. Functionally, SSEA-1 may participate in migration of the cells from neurospheres in an NPC cell culture system, and the effect could be repressed by anti-SSEA-1 antibody. Based on this observation, we identified beta1 integrin as one of the SSEA-1 carrier glycoproteins. Our data thus provide insights into the functional role of certain glycoconjugate antigens in NPCs during neural development. PMID- 16219036 TI - Frodo proteins: modulators of Wnt signaling in vertebrate development. AB - The Frodo/dapper (Frd) proteins are recently discovered signaling adaptors, which functionally and physically interact with Wnt and Nodal signaling pathways during vertebrate development. The Frd1 and Frd2 genes are expressed in dynamic patterns in early embryos, frequently in cells undergoing epithelial-mesenchymal transition. The Frd proteins function in multiple developmental processes, including mesoderm and neural tissue specification, early morphogenetic cell movements, and organogenesis. Loss-of-function studies using morpholino antisense oligonucleotides demonstrate that the Frd proteins regulate Wnt signal transduction in a context-dependent manner and may be involved in Nodal signaling. The identification of Frd-associated factors and cellular targets of the Frd proteins should shed light on the molecular mechanisms underlying Frd functions in embryonic development and in cancer. PMID- 16219037 TI - Xenopus connexins: how frogs bridge the gap. AB - Animal species use specialized cell-to-cell channels, called gap junctions, to allow for a direct exchange of ions and small metabolites between their cells' cytoplasm. In invertebrates, gap junctions are formed by innexins, while vertebrates use connexin (Cx) proteins as gap-junction-building blocks. Recently, innexin homologs have been found in vertebrates and named pannexins. From progress in the different genome projects, it has become evident that every class of vertebrates uses their own unique set of Cxs to build their gap junctions. Here, we review all known Xenopus Cxs with respect to their expression, regulation, and function. We compare Xenopus Cxs with those of zebrafish and mouse, and provide evidence for the existence of several additional, non identified, amphibian Cxs. Finally, we identify two new Xenopus pannexins by screening EST libraries. PMID- 16219038 TI - Developmental expression of two Haliotis asinina hemocyanin isoforms. AB - Hemocyanins are large copper-containing respiratory proteins that play a role in oxygen transport in many molluscs. In some species only one hemocyanin isoform is present while in others two are expressed. The physiological relevance of these isoforms is unclear and the developmental and tissue-specific expression of hemocyanin genes is largely unknown. Here we show that two hemocyanin genes in the gastropod Haliotis asinina, which encode H. asinina hemocyanin (HaH1) and HaH2 isoforms, are developmentally expressed. These genes initially are expressed in a small number of mesenchyme cells at trochophore and pre-torsional veliger stages, with HaH1 expression slightly preceding HaH2. These cells largely are localized to the visceral mass, although a small number of cells are present in head and foot regions. Following metamorphosis the isoforms show overlapping as well as isoform-specific expression profiles, suggesting some degree of isoform specific function. PMID- 16219039 TI - Quantitative transcriptional profiling of ATDC5 mouse progenitor cells during chondrogenesis. AB - During the differentiation of a mouse chondroprogenitor cell line, ATDC5, an analysis of the transcription cartilage-related genes was carried out using real time RT-PCR in a semiquantitative fashion. A total number of 104 genes both previously linked to chondrogenesis and hitherto not associated with the development of cartilage were analyzed. Parametric statistics, and unsupervised hierarchical and K-medians clustering approaches were used to analyze the gene expression during the sequential processes of proliferation, condensation, differentiation, maturation, and hypertrophic conversion of ATDC5 cells. The obtained data provided a robust determination of expression patterns that make possible an accurate assessment of the molecular events along the chondrogenic differentiation pathway. In addition, time-course expression profiles were described for eight highly regulated genes that have not been associated with chondrogenesis as yet. These included Cryab, Rcor2, Hig1, Bnip3, Mst4, Calml4, Gng2, and Islr. PMID- 16219040 TI - Ci-Rga, a gene encoding an MtN3/saliva family transmembrane protein, is essential for tissue differentiation during embryogenesis of the ascidian Ciona intestinalis. AB - A novel gene (Ci-Rga) essential for tissue differentiation during embryogenesis of the ascidian Ciona intestinalis is reported here. This gene was identified through functional screening of Ciona genes required for development by translational inhibition experiments with morpholino antisense oligonucleotides. The deduced protein of Ci-Rga contains two copies of a domain with unknown function called the MtN3/saliva domain. Phylogenetic analysis showed that Ci-Rga belongs to the MtN3/saliva family of genes conserved among metazoans and plants, and is an ortholog of mouse Rga (Recombination-activating gene 1 gene activation). During Ciona embryogenesis, both maternal and zygotic transcripts of Ci-Rga were expressed. Translational inhibition of Ci-Rga with specific morpholino resulted in abnormal embryos in which the cleavage pattern became atypical and expression of marker genes for each of the six major tissues, namely the endoderm, muscle, mesenchyme, notochord, neural tissue, and epidermis, was lost or suppressed at the tailbud stage. Although differentiation of all the six major tissues was affected by Ci-Rga knock-down, the degree of abnormalities and the timing of appearance of abnormalities were different among tissues. Expression analysis of developmentally important genes involved in the fate specification, such as Ci-Bra, Ci-Twist-like1a, Ci-Otx, Ci-Fgf9/16/20, Ci-Lhx3, Ci-FoxD, and Ci-Tbx6b, showed that an initial step of the fate specification of notochord, mesenchyme, and neural tissue, but not of endoderm or muscle, is impaired in the knock-down embryo. These results showed that Ci-Rga is a multifunctional gene essential for tissue differentiation during embryogenesis, and is primarily required for the fate specification of notochord, mesenchyme, and neural tissue, and provide some insights into the function of this little known group of genes. PMID- 16219041 TI - Dictyopyrones, novel alpha-pyronoids isolated from Dictyostelium spp., promote stalk cell differentiation in Dictyostelium discoideum. AB - Dictyopyrones A and B (DpnA and B), whose function(s) is not known, were isolated from fruiting bodies of Dictyostelium discoideum. In the present study, to assess their function(s), we examined the effects of Dpns on in vitro cell differentiation in D. discoideum monolayer cultures with cAMP. Dpns at 1-20 microM promoted stalk cell formation to some extent in the wild-type strain V12M2. Although Dpns by themselves could hardly induce stalk cell formation in a differentiation-inducing factor (DIF)-deficient strain HM44, both of them dose dependently promoted DIF-1-dependent stalk cell formation in the strain. In the sporogenous strain HM18, Dpns at 1-20 microM suppressed spore formation and promoted stalk cell formation in a dose-dependent manner. Analogs of Dpns were less effective in affecting cell differentiation in both HM44 and HM18 cells, indicating that the activity of Dpns should be chemical structure specific. It was also shown that DpnA at 2-20 microM dose-dependently suppressed spore formation induced with 8-bromo cAMP and promoted stalk cell formation in V12M2 cells. Interestingly, it was shown by the use of RT-PCR that DpnA at 10 microM slightly promoted both prespore- and prestalk-specific gene expressions in an early phase of V12M2 and HM18 in vitro differentiation. The present results suggest that Dpns may have functions (1) to promote both prespore and prestalk cell differentiation in an early stage of development and (2) to suppress spore formation and promote stalk cell formation in a later stage of development in D. discoideum. PMID- 16219042 TI - Tailoring haemostatic treatment to patient requirements - an update on monitoring haemostatic response using thrombelastography. AB - Currently, there is no single haemostasis laboratory test that has the capacity to accurately illustrate the clinical effects of procoagulant or anticoagulant interventions. Although the time course of thrombin generation in plasma and the endogenous thrombin potential (ETP) may be useful coagulation parameters, clotting involves components other than thrombin (e.g. platelets, fibrinogen). The continuous coagulation profiles of thrombelastography may provide a more accurate reflection of in vivo biology, covering initiation, development and final clot strength during whole blood clot formation. This method has helped to clarify the mechanism of action of whole blood clot formation, demonstrating the differences from clotting in plasma, and the importance of platelets and tissue factor titrations. It has also been used to investigate hypocoagulation (in haemophilia A, rare coagulation disorders, anticoagulant therapy and dilutional coagulopathy), hypercoagulation and the ex vivo testing of haemostatic interventions. Thrombelastography has been shown to reflect the clinical efficacy of activated prothrombin complex concentrate (aPCC) and recombinant activated factor VII (rFVIIa) in patients with haemophilia A with inhibitors and in patients with acquired haemophilia. Overall, tailoring laboratory assays to illustrate and correlate with clinical phenotypes is essential for effective coagulation monitoring. Applying an algorithm of preoperative, perioperative and postoperative tests, including thrombelastography, may enable physicians to achieve this. PMID- 16219043 TI - Identifying and managing inhibitor patients requiring orthopaedic surgery - the multidisciplinary team approach. AB - Until recently, surgery in haemophilia patients with inhibitors was strongly contraindicated and was therefore often not even contemplated. Inhibitor patients entering the surgical arena face unique challenges; the most frequently encountered problem during surgical intervention is bleeding, and thrombosis is occasionally observed. The activated prothrombin complex concentrate, FEIBA(TM), and recombinant activated factor VII (rFVIIa) are available as haemostatic cover during surgery. The use of rFVIIa enables inhibitor patients to undergo orthopaedic surgery with an expectation of success, and results are generally good. An organized team approach is critical to this success. However, further information is required to enable different procedures to be optimized in terms of both outcome and safety. PMID- 16219044 TI - Health economics of treating haemophilia A with inhibitors. AB - Haemophilia is a rare, inherited blood disorder in which blood clotting is impaired such that patients suffer from excessive internal and external bleeding. At present there is no cure for haemophilia A and patients require expensive, life-long treatment involving clotting factor replacement therapy. Treatment costs are perceived to be higher for patients who have developed inhibitory antibodies to factor VIII, the standard therapy for haemophilia A. However, initial cost analyses suggest that clotting factor therapy with alternative haemostatic agents, such as recombinant activated factor VII or activated prothrombin complex concentrate, is no more expensive for the majority of haemophilia A patients with inhibitors than for those without inhibitors. With the availability of effective alternative haemostatic agents, orthopaedic surgery for haemophilia A patients with inhibitors is now a clinical option, and initial cost analyses suggest this may be a cost-effective treatment strategy for patients with inhibitors whose quality of life (QoL) is severely impaired by joint arthropathy. In an era of finite healthcare resourcing it is important to determine whether new treatments justify higher unit costs compared with standard therapies and whether such higher costs are justified from an individual perspective in terms of improved QoL, and from a societal perspective in terms of improved productivity and reduced overall healthcare costs. This paper examines current data on the health economics of treating haemophilia A patients with inhibitors, focusing on the overall costs of clotting factor replacement therapy and the cost consequences of joint replacement. PMID- 16219045 TI - Managing acute bleeds in the patient with haemophilia and inhibitors: options, efficacy and safety. AB - The options available for treating the patient with haemophilia and inhibitors undergoing surgery or with other acute bleeds include high-dose factor VIII (FVIII) (human or porcine), prothrombin complex concentrates (PCCs), activated PCCs (aPCCs), recombinant activated factor VII (rFVIIa), and factor replacement combined with immunoadsorption or immunosuppression. Human FVIII is effective in patients with low-titre inhibitors. Porcine FVIII is currently not available, and PCCs and aPCCs, although effective, have been associated with a high incidence of adverse events. Immunoadsorption and immunosuppression offer excellent long-term solutions, but the duration of these techniques makes them less attractive for use in acute settings. Recombinant FVIIa has demonstrated excellent efficacy and safety, even in patients refractory to other therapies. PMID- 16219046 TI - ESOS: a European study on the orthopaedic status of patients with haemophilia and inhibitors. AB - Published studies highlight the paucity of data relating to orthopaedic health and quality of life in haemophilia patients with inhibitors. A European Study on the Orthopaedic Status of Patients with Haemophilia and Inhibitors (ESOS) has been set up to address this lack of data. This cross-sectional retrospective study aims to enrol 400 patients from 50 sites in nine European countries. The primary outcome measure of the study is the burden of orthopaedic complications in patients with severe haemophilia and inhibitors. The data that will be generated should demonstrate the high level of arthropathy in haemophilia patients with inhibitors and show the need to focus on improving orthopaedic health in this rare, but particularly vulnerable, patient population. PMID- 16219047 TI - Rehabilitation of patients with haemophilia after orthopaedic surgery: a case study. AB - Preoperative and postoperative rehabilitation may be useful for improving the recovery of patients undergoing orthopaedic surgery, particularly in those with co-morbidity or special requirements. This case study, of a patient with haemophilia and inhibitors to factor VIII undergoing total knee replacement, demonstrates the benefits of 6 weeks' preoperative physiotherapy ('prehabilitation') combined with 6 weeks' postoperative rehabilitation. The supervised physiotherapy regimen was individually tailored to specifically increase range of motion and muscle strength, enabling rapid mobilization and recovery of function, whilst minimizing the risk of bleeding. PMID- 16219048 TI - Quality of life before and after surgery: mobility issues, the fear of surgery, inpatient recovery and outpatient rehabilitation. AB - A positive life-changing experience of joint replacement surgery from the point of view of a patient with Factor VII inhibitor. PMID- 16219049 TI - Case studies: orthopaedic surgery in adult patients with haemophilia A with inhibitors. AB - Whilst orthopaedic surgery in haemophilia patients without inhibitors is now relatively common in specialized centres, until recently there have been only a few sporadic instances of surgery having been undertaken on patients with inhibitors. The availability of recombinant activated factor VII (rFVIIa) for haemostatic cover during surgery allows procedures to be performed that previously may not have been considered possible. Complications associated with thrombosis are rare in haemophilia patients with inhibitors, but bleeding complications remain a concern. Globally, experience of performing orthopaedic surgery in these patients is increasing and many successful outcomes have been reported. However, more knowledge relating to the incidence and type of bleeding complications liable to be encountered, together with further information about appropriate rescue treatment, would be valuable. Data relating to long-term follow-up after surgery would be useful, as would a comparison of outcomes between haemophilia patients with and without inhibitors. Optimal dosing regimens for rFVIIa as surgical cover are still to be determined and further information is required relating to the cost effectiveness of rFVIIa in surgery. Further study should address these issues. PMID- 16219050 TI - Indices for assessment of hemodialysis adequacy: a comparison of different formulae. AB - Of the various indices used in the assessment of dialysis adequacy, fractional urea clearance controlled for volume of distribution "Kt/V" remains the most widely used. Its determination is best performed by formal urea kinetic modeling (UKM), which is laborious and cumbersome, and the computational softwares are largely unavailable, particularly in developing countries. Consequently, different equations have been developed that approximate the formal UKM determination. Of the available formulae, that from second-generation logarithmic equation have been found to approximate values derived from formal UKM closely. We set out to determine the clinical utility of percent reduction of urea and Kt/V formulae derived from it, using the logarithmic equation as the standard. PMID- 16219051 TI - Factors influencing low-molecular-weight solute clearance during hemodialysis. AB - The relationship between delivered urea Kt/V and survival has raised concerns in recent outcome trials in chronic dialysis patients. Nevertheless, measurement of delivered small solute clearance remains the most common approach to quantify therapy. The purpose of this review is to provide an overview of the numerous factors influencing small solute clearance during hemodialysis. Although the focus of the review is on the manner in which dialyzer characteristics influence small solute clearances, factors related to other aspects of the extracorporeal circuit and to the patient will also be discussed. PMID- 16219052 TI - Effect of mineralocorticoids on interdialytic weight gain in hemodialysis patients with perdialytic hypotension. AB - Fludrocortisone is recommended in patients with orthostatic hypotension and a benefit has been suggested in hemodialysis patients with severe hypokaliemia. We report 2 patients who suffered from chronic severe perdialytic hypotension resistant to midodrine and who were treated in a long-term period with fludrocortisone. A rise of post-dialytic BP was observed with a decrease of the interdialytic weight gain (IWG). We suggest that the IWG decrease is induced by a lessening of the renin angiotensin aldosterone system that could be less stimulated at the end of the dialysis session because of a better-preserved arterial pressure. The decrease of angiotensin could lessen the feeling of thirst. PMID- 16219053 TI - Accidental extravascular insertion of a subclavian hemodialysis catheter is signaled by nonvisualization of catheter tip. AB - Subclavian hemodialysis (HD) catheter placement under fluoroscopy with perforation of the superior vena cava (SVC) is a rare complication that needs to be recognized and treated appropriately. We report the case of a 47-year-old black woman under treatment for end-stage renal disease secondary to HIV associated nephropathy who sustained an extravascular insertion of fluoroscopy guided subclavian catheterization for HD. Subsequent successful removal of the extravascularly placed catheter along with repair of the lacerated SVC were effected by open thoracic surgery. PMID- 16219054 TI - Reflex sympathetic dystrophy syndrome due to arteriovenous fistula. AB - A patient with end-stage renal disease presented with reflex sympathetic dystrophy syndrome (RSDS) on her left hand 1 month after arteriovenous fistula (AVF) surgery. Magnetic resonance angiography revealed steal syndrome at the AVF level. Bone scintigraphy revealed early-stage RSDS. We considered that arterial insufficiency because of steal phenomenon following AVF surgery and underlying occlusive arterial disease triggered RSDS development. PMID- 16219055 TI - The morbidity and cost implications of hemodialysis clinical performance measures. AB - Clinical performance measures, including dialysis dose, hemoglobin, albumin, and vascular access, are the focus of monitoring and quality improvement activities. However, little is known about the implications of clinical performance measures for hospital utilization and health care costs. We obtained clinical performance measures and hospitalization records for a national random sample of 10,650 hemodialysis patients and analyzed the relationship between changes in clinical performance measures and hospital utilization after adjustment for patient demographic and medical characteristics. Higher hemoglobin, higher albumin, and fistula or graft use were independently associated with fewer hospitalizations, fewer hospital days, and decreased Medicare inpatient reimbursement. For example, a 0.5 g/dL higher hemoglobin, a 0.25 g/dL higher albumin, fistula use, and graft use were associated with hospitalization rate ratios of 0.90 (95% confidence interval 0.85, 0.96), 0.64 (0.53, 0.77), 0.60 (0.52, 0.69), and 0.79 (0.71, 0.89), respectively. Moreover, there was a 2-3-fold variation in hospital utilization across end-stage renal disease networks that was still evident after adjustment for patient characteristics and clinical performance measures. Clinical performance measures, especially albumin and vascular access, are strongly associated with hospital utilization and health care costs. These results highlight the importance of targeting nutrition and vascular access in quality improvement efforts. The marked variation in hospital utilization across networks deserves further examination. PMID- 16219056 TI - Prevalence of transfusion-transmitted virus infection in patients on maintenance hemodialysis from New Delhi, India. AB - Transfusion-transmitted virus (TTV) has been reported from a number of hemodialysis (HD) units from various countries throughout the world. TTV has been associated with liver diseases, viral hepatitis B, and C. Clinical details and information regarding TTV prevalence from India are insufficient. The prevalence and clinical significance of TTV infection were studied in New Delhi, India in HD patients. Serum samples were derived from 75 patients on maintenance HD, and 75 age- and sex-matched voluntary blood donors were examined for TTV viremia by nested polymerase chain reaction (PCR) using primers derived from UTR (A) region of the TTV genome. The prevalence of TTV DNA in patients on HD (83%) was significantly (p<0.05) higher than in blood donors (43%). Clinical background including the mean age, sex, mean duration of HD, and mean alanine aminotransferase (ALT) levels did not differ significantly between TTV DNA positive and -negative HD patients. Fifty-four (72%) TTV-positive HD patients and 7 (56%) TTV-negative HD patients had blood transfusion histories (p>0.05). Among TTV-positive patients, Hepatitis B virus (HBV) co-infection was present in 14.2% cases while hepatitis C virus (HCV) co-infection was absent. Persistent elevation of ALT levels was observed in 7(9.3%) HD patients; 3 (43%) of them were TTV positive and 4 (57%) were TTV negative (p>0.05). All 3 TTV-positive patients with elevated ALT levels were co-infected with HBV. Patients with TTV infection alone showed normal ALT levels. Prevalence of TTV infection is high in North Indian patients on maintenance HD. Also, none of the exclusively TTV DNA-positive patients had clinical or biochemical signs of liver disease. TTV seems to spread through parenteral routes. More often, TTV seems to be associated with parenterally transmitted virus HBV, indicating a parenteral mode of TTV transmission. The pathogenicity of TTV remains unclear from the present study. PMID- 16219057 TI - Antibody response to Engerix-B and Recombivax-HB hepatitis B vaccination in end stage renal disease. AB - Recombivax-HB (REC) and Engerix-B (ENG) are FDA-approved vaccines for hepatitis B virus (HBV) in end-stage renal disease (ESRD). This study compares antibody response rates between them in routine clinical practice. Patients completing the recommended 40 mug dose of REC (3 doses) or ENG (4 doses) between January 1, 2000 to April 30, 2003 were eligible. Patients with prior positive HBV surface antigen (HBsAg) or antibody (HBsAb) test results were excluded. The conversion rate and persistence of protective titer (HBsAb titer>or=10 IU/mL) were tracked for 1 year. A supplemental analysis of a one-to-one matched patient sample was also performed. REC patients (N=885) were older, had longer dialysis vintage, and had a larger proportion of whites than ENG patients (N=13,661). Cumulative conversion response was greater in ENG (58%) than REC (40%) at 1 year (p<0.0001). The odds ratio for response to ENG compared with REC was 1.96 (95% limits: 1.56, 2.45; p<0.0001) adjusted for age, gender, race, diabetes, vintage, BSA, hemoglobin, and eKt/V. Persistent protective HBsAb after 1 year was 77% (ENG) vs. 53% (REC). HBsAg was positive in 208 ENG patients (1.5%) with all but 1 because of transient, vaccine-related antigenemia. The difference in conversion response favoring ENG persisted in a one-to-one sample matched for age, gender, race, modality, and dialysis vintage. The study found higher seroconversion response to ENG compared with REC at several time points up to 1 year. Protective HBsAb disappeared in 23-47% of patients 1 year later, validating CDC recommendations to re-test HBsAb yearly. The observed difference in response rates may be related to the extra ENG dose given at the second month (0, 1, 2, 6 regimen). The study raises a hypothesis that requires confirmation in a prospective clinical trial. PMID- 16219058 TI - Intradialytic changes in reflective properties of the arterial system during a single hemodialysis session. AB - Increased aortic stiffness-measured as aortic augmentation index (AIx), a global stiffness marker-has emerged as a powerful predictor of survival in hemodialysis (HD). A single HD session is known to produce considerable improvement in aortic stiffness. We set out, for the first time, to examine the relative contributions to the post-HD drastic improvement in aortic stiffness of ultrafiltration rate and volume, or blood pressure (BP) changes. Aortic AIx (difference between the first and the second systolic peak of the aortic pressure waveform divided by pulse wave height) was determined hourly and recorded by applanation tonometry using a SphygmoCor device in 20 chronic HD patients (9 males, age 55.1 years). The other parameters recorded were: weight pre- and post-HD, ultrafiltration volume (UFV), hemoglobin, albumin, creatinine, urea reduction rate (URR), calcium and PTH, and BP. The dialysis significantly decreased AIx from 24.2+/-11.27% to 15.57+/-12.58% (p<0.05). In a univariate analysis, the intradialytic decrease in AIx (AIx 0-4) did not correlate with UFV, URR or with any of the biochemical markers. Significant correlations for AIx 0-4 were age (p=0.018), systolic blood pressure (SBP) at the beginning of HD (p=0.049), the intradialytic decrease in the SBP (p=0.001), and in pulse pressure (PP) (p=0.009). Multivariate stepwise regression showed that the decrease in SBP, PP, and intradialytic percentage reduction in weight explained 64.9% of the total variation in AIx 0-4. The decrease in SBP was the most important factor influencing the AIx variation (b=1.54, p=0.007). The most significant reduction in AIx was from the beginning of HD to the third hour (p=0.039), and correlated with the reduction in SBP (p=0.006) and PP (p=0.025) between the same moments. A single HD session produces a drastic improvement in aortic stiffness. The effect is not explained by the UFV depletion but is highly correlated with the decrease in SBP and PP. Further work is now needed to explore a potential role for endothelin and nitric oxide metabolism. PMID- 16219059 TI - Automatic feedback control of relative blood volume changes during hemodialysis improves blood pressure stability during and after dialysis. AB - Automatic feedback systems have been designed to control relative blood volume changes during hemodialysis (HD) as hypovolemia plays a major role in the development of dialysis hypotension. Of these systems, one is based on the concept of blood volume tracking (BVT). BVT has been shown to improve intra-HD hemodynamic stability. We first questioned whether BVT also improves post-HD blood pressure stability in hypotension-prone patients and second, whether BVT is effective in reducing the post-HD weight as many hypotension-prone patients are overhydrated because of an inability to reach dry weight. After a 3-week period on standard HD, 12 hypotension-prone patients were treated with two consecutive BVT treatment protocols. During the first BVT period of 3 weeks, the post-HD target weight was kept identical compared with the standard HD period (BVT constant weight; BVT-cw). During the second BVT period of 6 weeks, we gradually tried to lower the post-HD target weight (BVT-reduced weight; BVT-rw). In the last week of each period, we studied intra-HD and 24 hr post-HD blood pressure behavior by ambulatory blood pressure measurement (ABPM). Pre- and post-HD weight did not differ between standard HD and either BVT-cw or BVT-rw. Heart size on a standing pre-dialysis chest X-ray did not change significantly throughout the study. There were less episodes of dialysis hypotension during BVT compared with standard HD (both BVT periods: p<0.01). ABPM data were complete in 10 patients. During the first 16 hr post-HD, systolic blood pressure was significantly higher with BVT in comparison with standard HD (both BVT periods: p<0.05). The use of BVT in hypotension-prone patients is associated with higher systolic blood pressures for as long as 16 hr post-HD. BVT was not effective in reducing the post-HD target weight in this patient group. PMID- 16219060 TI - Clinical consequences of heparin-free hemodialysis. AB - Heparin-free hemodialysis (HF-HD) has been increasingly used in patients at risk for bleeding, especially in the intensive care unit (ICU). Lack of heparin can reduce solute clearances in continuous hemofiltration; the effect on HD is undefined. Failure to recognize an effect of the anticoagulation strategy upon delivered clearance could contribute to the known problem of underdialysis in the ICU. In addition, the consequences of "locking" dialysis catheters with concentrated heparin solutions are also unclear. This study was designed to define the clinically relevant consequences of HF-HD and catheter locking. In part I, we performed 200 HD treatments on inpatients, of which 100 were performed with heparin, and 100 were performed as HF-HD. We calculated prescribed and delivered Kt/V and dialysis efficiency. In part II, a separate group of 14 patients undergoing HF-HD via central venous catheters had measurement of activated partial thromboplastin time (aPTT) during the last hour of dialysis, as well as 15, 60, and 240 min after catheters were locked with 1:5000 heparin. The prescribed Kt/V was 1.74+/-0.31 for standard HD with heparin vs. 1.66+/-0.36 for HF-HD (p=ns). The delivered Kt/V was 1.42+/-0.32 vs. 1.36+/-0.38 (p=ns). Efficiency was 0.82 vs. 0.84 (p=ns). Baseline aPTT was 28+/-5 s, and increased to 126+/-54 s, 15 min after locking (p<0.0001) and to 71+/-50 s, 60 min after locking (p=0.005). By 240 min, the mean aPTT had fallen to 33+/-9 s (p=0.03), although individual values were still as high as 50 s. The HF technique does not compromise delivery of dialysis to inpatients. Increased treatment time is not necessary. Locking catheters with heparin after HF-HD resulted in prolonged unintentional anticoagulation. PMID- 16219061 TI - Middle molecule removal in low-flux polysulfone dialyzers: impact of flows and surface area on whole-body and dialyzer clearances. AB - Some studies found that the removal of middle molecules has a long-term effect on mortality and, even more, is enhanced by high-flux dialysis. In order to enhance middle molecule removal in a low-flux dialyzer, the present study aimed at investigating the combined impact of dialyzer flows and membrane surface area. Blood and dialysate flows were varied within the clinical range 300-500 and 500 800 mL/min, respectively, while the ultrafiltration rate was kept constant at 0.1 L/hr. Single-pass tests were performed in vitro in a single Fresenius F6HPS dialyzer (3 tests) and serially (5 tests) and parallel (3 tests) connected dialyzers. The blood substitution fluid consisted of dialysis fluid in which radioactive-labeled vitamin B12 (molecular weight 1355 Da) was dissolved. Dialyzer clearance as well as whole-body clearance was calculated from radioactivity concentrations of samples taken from the inlet and outlet bloodline. Adding a second dialyzer in series or parallel ameliorated the overall dialyzer and whole-body clearance significantly, except for the highest applied blood flows of 500 mL/min. Better solute removal was also obtained with higher dialysate flows, while the use of higher blood flows seemed advantageous only when using a single dialyzer. Analysis of the ultrafiltration profiles in the different configurations illustrated that enhancing the internal filtration rate ameliorates convective transport of middle molecules. Adequate solute removal results from a number of interactions, as there are blood and dialysate flows, membrane surface area, filtration profile and concentration profiles in the blood and dialysate compartment. PMID- 16219062 TI - Hyperphosphatemia among end-stage renal disease patients in developing countries: a forgotten issue? AB - The prevalence of hyperphosphatemia and increased calcium-phosphorus product has never been evaluated in a large multicenter study in a developing country. Our aim is to study the prevalence of hyperphosphatemia in 38 HD centers in Egypt (as an example of a developing country) and to correlate it with different co-morbid conditions and the patient's demographic data. This is a cross-sectional study conducted on 1005 chronic kidney disease stage 5 patients (CKD-stage 5) on HD for a period of more than 1 year in 38 dialysis centers in Egypt. All patients were receiving calcium-based salts as a phosphate binder. Hyperphosphatemia and increased calcium-phosphorus product were evaluated and correlated with different parameters including age, sex, knowledge by diet parameters, HD session duration, the frequency of HD per week, the type of dialysis membrane, the surface area of the dialyzer, dialyzer phosphorus clearance (phosphorus KoA), and the type of dialysate. Other co-morbid medical conditions and evidence of IHD were also investigated. Hyperphosphatemia was present in 69.1% of cases and a high calcium phosphorus product was present in 30.2%. A higher calcium-phosphorus product was found among males. 83.2% of those with a poor knowledge by diet parameters had hyperphosphatemia compared with 67.6% in patients with a satisfactory knowledge by diet parameters. 72.3% of patients using a membrane with low-to-medium clearance had hyperphosphatemia, compared with 67.2% using a membrane with a high clearance. Seventy-two percent of patients with IHD were hyperphosphatemic compared with 67.6% of the non-ischemic patients. Hyperphosphatemia is a major problem in dialysis patients in developing countries, reflecting differences from developed countries regarding dietary habits, ethnic factors, dialysis quality, types of dialysis membranes, as well as economic factors hampering the use of the more expensive phosphate binders. Extended dialysis hours may be a good alternative solution in developing countries. PMID- 16219063 TI - "Uremic pruritus": a misnomer. PMID- 16219064 TI - Global biogeography of plant chemistry: filling in the blanks. PMID- 16219065 TI - Towards a multifunctional rhizosphere concept: back to the future? PMID- 16219066 TI - Measuring quality of service: phosphate 'a la carte' by arbuscular mycorrhizal fungi. PMID- 16219067 TI - Why don't leaf-eating animals prevent the formation of vegetation? Relative vs absolute dietary requirements. PMID- 16219068 TI - The control of stomata by water balance. AB - It is clear that stomata play a critical role in regulating water loss from terrestrial vegetation. What is not clear is how this regulation is achieved. Stomata appear to respond to perturbations of many aspects of the soil-plant atmosphere hydraulic continuum, but there is little agreement regarding the mechanism (or mechanisms) by which stomata sense such perturbations. This review discusses feedback and feedforward mechanisms by which hydraulic perturbations are putatively transduced into stomatal movements, in relation to generic empirical features of those responses. It is argued that a metabolically mediated feedback response of stomatal guard cells to the water status in their immediate vicinity ('hydro-active local feedback') remains the best explanation for many well-known features of hydraulically related stomatal behaviour, such as transient 'wrong-way' responses and the equivalence of hydraulic supply and demand as stomatal effectors. Furthermore, many curious phenomena that appear inconsistent with feedback, such as 'apparent feedforward' humidity responses and 'isohydric' behaviour (water potential homeostasis), are in fact expected to emerge from the juxtaposition of hydro-active local feedback and the well-known hysteretic and threshold-like effect of water potential on xylem hydraulic resistance. PMID- 16219069 TI - Rhizosphere geometry and heterogeneity arising from root-mediated physical and chemical processes. AB - The rhizosphere differs from the bulk soil in a range of biochemical, chemical and physical processes that occur as a consequence of root growth, water and nutrient uptake, respiration and rhizodeposition. These processes also affect microbial ecology and plant physiology to a considerable extent. This review concentrates on two features of this unique environment: rhizosphere geometry and heterogeneity in both space and time. Although it is often depicted as a soil cylinder of a given radius around the root, drawing a boundary between the rhizosphere and bulk soil is an impossible task because rhizosphere processes result in gradients of different sizes. For instance, because of diffusional constraints, root uptake can result in a depletion zone extending <1 mm for phosphate to several centimetres for nitrate, while respiration may affect the bulk of the soil. Rhizosphere processes are responsible for spatial and temporal heterogeneities in the soil, although these are sometimes difficult to distinguish from intrinsic soil heterogeneity. A further complexity is that these processes are regulated by plants, microbial communities and soil constituents, and their many interactions. Novel in situ techniques and modelling will help in providing a holistic view of rhizosphere functioning, which is a prerequisite for its management and manipulation. PMID- 16219070 TI - Nutrient availability and management in the rhizosphere: exploiting genotypic differences. AB - Crop nutrition is frequently inadequate as a result of the expansion of cropping into marginal lands, elevated crop yields placing increasing demands on soil nutrient reserves, and environmental and economic concerns about applying fertilizers. Plants exposed to nutrient deficiency activate a range of mechanisms that result in increased nutrient availability in the rhizosphere compared with the bulk soil. Plants may change their root morphology, increase the affinity of nutrient transporters in the plasma membrane and exude organic compounds (carboxylates, phenolics, carbohydrates, enzymes, etc.) and protons. Chemical changes in the rhizosphere result in altered abundance and composition of microbial communities. Nutrient-efficient genotypes are adapted to environments with low nutrient availability. Nutrient efficiency can be enhanced by targeted breeding through pyramiding efficiency mechanisms in a desirable genotype as well as by gene transfer and manipulation. Rhizosphere microorganisms influence nutrient availability; adding beneficial microorganisms may result in enhanced availability of nutrients to crops. Understanding the role of plant-microbe-soil interactions in governing nutrient availability in the rhizosphere will enhance the economic and environmental sustainability of crop production. PMID- 16219071 TI - The dark side of green fluorescent protein. AB - Here, severe interference of chlorophyll with green fluorescent protein (GFP) fluorescence is described for medicago (Medicago truncatula), rice (Oryza sativa) and arabidopsis (Arabidopsis thaliana). This interference disrupts the proportional relationship between GFP content and fluorescence that is intrinsic to its use as a quantitative reporter. The involvement of chlorophyll in the loss of GFP fluorescence with leaf age was shown in vivo, by the removal of chlorophyll through etiolation or by ethanol extraction, and in vitro, by titration of a GFP solution with chlorophyll solutions of various concentrations. A substantial decrease in fluorescence in early development of medicago and rice leaves correlated with chlorophyll accumulation. In all three species tested, removal of chlorophyll yielded up to a 10-fold increase in fluorescence. Loss of GFP fluorescence in vitro was 4-fold greater for chlorophyll b than for chlorophyll a. Differences exist between plant species for the discrepancy between apparent GFP fluorescence and its actual level in green tissues. Substantial errors in estimating promoter activity from GFP fluorescence can occur if pigment interference is not considered. PMID- 16219073 TI - Nickel hyperaccumulation as an elemental defense of Streptanthus polygaloides (Brassicaceae): influence of herbivore feeding mode. AB - No study of a single nickel (Ni) hyperaccumulator species has investigated the impact of hyperaccumulation on herbivores representing a variety of feeding modes. Streptanthus polygaloides plants were grown on high- or low-Ni soils and a series of no-choice and choice feeding experiments was conducted using eight arthropod herbivores. Herbivores used were two leaf-chewing folivores (the grasshopper Melanoplus femurrubrum and the lepidopteran Evergestis rimosalis), a dipteran rhizovore (the cabbage maggot Delia radicum), a xylem-feeder (the spittlebug Philaenus spumarius), two phloem-feeders (the aphid, Lipaphis erysimi and the spidermite Trialeurodes vaporariorum) and two cell-disruptors (the bug Lygus lineolaris and the whitefly Tetranychus urticae). Hyperaccumulated Ni significantly decreased survival of the leaf-chewers and rhizovore, and significantly reduced population growth of the whitefly cell-disruptor. However, vascular tissue-feeding insects were unaffected by hyperaccumulated Ni, as was the bug cell-disruptor. We conclude that Ni can defend against tissue-chewing herbivores but is ineffective against vascular tissue-feeding herbivores. The effects of Ni on cell-disruptors varies, as a result of either variation of insect Ni sensitivity or the location of Ni in S. polygaloides cells and tissues. PMID- 16219072 TI - The novel herbicide oxaziclomefone inhibits cell expansion in maize cell cultures without affecting turgor pressure or wall acidification. AB - Oxaziclomefone [OAC; IUPAC name 3-(1-(3,5-dichlorophenyl)-1-methylethyl)-3,4 dihydro-6-methyl-5-phenyl-2H-1,3-oxazin-4-one] is a new herbicide that inhibits cell expansion in grass roots. Its effects on cell cultures and mode of action were unknown. In principle, cell expansion could be inhibited by a decrease in either turgor pressure or wall extensibility. Cell expansion was estimated as settled cell volume; cell division was estimated by cell counting. Membrane permeability to water was measured by a novel method involving simultaneous assay of the efflux of (3)H(2)O and [(14)C]mannitol from a 'bed' of cultured cells. Osmotic potential was measured by depression of freezing point. OAC inhibited cell expansion in cultures of maize (Zea mays), spinach (Spinacia oleracea) and rose (Rosa sp.), with an ID(50) of 5, 30 and 250 nm, respectively. In maize cultures, OAC did not affect cell division for the first 40 h. It did not affect the osmotic potential of cell sap or culture medium, nor did it impede water transport across cell membranes. It did not affect cells' ability to acidify the apoplast (medium), which may be necessary for 'acid growth'. As OAC did not diminish turgor pressure, its ability to inhibit cell expansion must depend on changes in wall extensibility. It could be a valuable tool for studies on cell expansion. PMID- 16219074 TI - Chromosomal regions with quantitative trait loci controlling cadmium concentration in brown rice (Oryza sativa). AB - A novel mapping population consisting of 39 chromosome segment substitution lines (CSSLs) was used to locate the putative quantitative trait loci (QTLs) for cadmium (Cd) concentration in brown rice (Oryza sativa). The mapping population carried a single chromosome segment of 'Kasalath' (indica) in each line overlapping with neighbouring segments in a 'Koshihikari' (japonica) genetic background. The parents and CSSLs were grown in pots filled with Cd-polluted soil until grain filling. The brown rice of three of the 39 CSSLs had significantly lower Cd concentrations than that of Koshihikari, and the brown rice of a further three had significantly higher concentrations. On the basis of graphical genotypes of CSSLs, putative QTLs controlling the Cd concentration in brown rice were detected on chromosomes 3, 6 and 8. Each of the CSSLs was nearly isogenic to Koshihikari, which is the most popular rice cultivar in Japan: they carried > 90% of the Koshihikari genetic background. Therefore, the development of a new Koshihikari with less Cd concentration in brown rice would be feasible in the near future. PMID- 16219075 TI - Differences in the kinetics and scale of signalling molecule production modulate the ozone sensitivity of hybrid poplar clones: the roles of H2O2, ethylene and salicylic acid. AB - Hydrogen peroxide (H(2)O(2)), ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC) and salicylic acid (SA) concentrations and ACC synthase (ACS) gene expression were measured to establish whether the high sensitivity of the Populus deltoides x maximowiczii clone Eridano to ozone (O(3)) exposure, compared with the O(3)-resistant Populus deltoides x euramericana clone I-214, is attributable to differences in the modulation of signal transduction pathways. In a time course experiment, Populus deltoides (poplar) clones were exposed to acute fumigation with 150 nl l(-1) O(3) for 5 h. The two poplar clones showed differences in ethylene evolution, I-214 displaying earlier and less pronounced ethylene emission than Eridano. In both clones, ethylene evolution was accompanied by increased ACS transcript levels and enhanced emission of free ACC. I-214 exhibited a greater basal concentration of free SA and a lower concentration of the conjugated pool. However, a slight accumulation of free SA at the end of the 5-h exposure was found only in Eridano, together with an earlier minimal increase in the concentration of conjugated SA. The results show that both clones react to O(3) by producing H(2)O(2), ethylene and SA, but the difference in sensitivity to the pollutant is probably attributable to differences in the kinetics and magnitude of this response. PMID- 16219076 TI - Stratification by cyanobacteria in lakes: a dynamic buoyancy model indicates size limitations met by Planktothrix rubescens filaments. AB - The ability of the Planktothrix rubescens to stratify in Lake Zurich is related to the size and shape of the cyanobacterial filaments. Detailed measurements made in the lake are used in a dynamic computer model of buoyancy regulation to investigate the vertical movements of filaments tracking the depth at which the irradiance would support neutral buoyancy. The movement of the filament lags behind the constantly changing target depth owing to (a) the time taken for the filament to respond to the irradiance by changing its density and (b) the time it takes to move by sinking down or floating up through the water column. The model simulates the stratification depth over a 5-month period of the summer from the continuous measurements of irradiance and weekly measurements of light attenuation and temperature, without any further adjustment over the period. Models using filaments of the size observed in Lake Zurich explain several details of the observed depth changes: smaller planktonic cyanobacteria (e.g. Limnothrix sp.) are unable to migrate fast enough and larger ones (e.g. Anabaena spp.) will overshoot and become entrained in the epilimnion. The model can be used to simulate recruitment of Planktothrix filaments from different depths after vernal stratification. Recruitment of filaments from depths down to 45 m will contribute to the metalimnetic population increase in early July. PMID- 16219077 TI - Leaf nitrogen and phosphorus stoichiometry across 753 terrestrial plant species in China. AB - Leaf nitrogen and phosphorus stoichiometry of Chinese terrestrial plants was studied based on a national data set including 753 species across the country. Geometric means were calculated for functional groups based on life form, phylogeny and photosynthetic pathway, as well as for all 753 species. The relationships between leaf N and P stoichiometric traits and latitude (and temperature) were analysed. The geometric means of leaf N, P, and N : P ratio for the 753 species were 18.6 and 1.21 mg g(-1) and 14.4, respectively. With increasing latitude (decreasing mean annual temperature, MAT), leaf N and P increased, but the N : P ratio did not show significant changes. Although patterns of leaf N, P and N : P ratios across the functional groups were generally consistent with those reported previously, the overall N : P ratio of China's flora was considerably higher than the global averages, probably caused by a greater shortage of soil P in China than elsewhere. The relationships between leaf N, P and N : P ratio and latitude (and MAT) also suggested the existence of broad biogeographical patterns of these leaf traits in Chinese flora. PMID- 16219078 TI - Root architecture and wind-firmness of mature Pinus pinaster. AB - This study aims to link three-dimensional coarse root architecture to tree stability in mature timber trees with an average of 1-m rooting depth. Undamaged and uprooted trees were sampled in a stand damaged by a storm. Root architecture was measured by three-dimensional (3-D) digitizing. The distribution of root volume by root type and in wind-oriented sectors was analysed. Mature Pinus pinaster root systems were organized in a rigid 'cage' composed of a taproot, the zone of rapid taper of horizontal surface roots and numerous sinkers and deep roots, imprisoning a large mass of soil and guyed by long horizontal surface roots. Key compartments for stability exhibited strong selective leeward or windward reinforcement. Uprooted trees showed a lower cage volume, a larger proportion of oblique and intermediate depth horizontal roots and less wind oriented root reinforcement. Pinus pinaster stability on moderately deep soils is optimized through a typical rooting pattern and a considerable structural adaptation to the prevailing wind and soil profile. PMID- 16219079 TI - Root proliferation and seed yield in response to spatial heterogeneity of below ground competition. AB - Here, we tested the predictions of a 'tragedy of the commons' model of below ground plant competition in annual plants that experience spatial heterogeneity in their competitive environment. Under interplant competition, the model predicts that a plant should over-proliferate roots relative to what would maximize the collective yield of the plants. We predict that a plant will tailor its root proliferation to local patch conditions, restraining root production when alone and over-proliferating in the presence of other plants. A series of experiments were conducted using pairs of pea (Pisum sativum) plants occupying two or three pots in which the presence or absence of interplant root competition was varied while nutrient availability per plant was held constant. In two-pot experiments, competing plants produced more root mass and less pod mass per individual than plants grown in isolation. In three-pot experiments, peas modulated this response to conditions at the scale of individual pots. Root proliferation in the shared pot was higher compared with the exclusively occupied pot. Plants appear to display sophisticated nutrient foraging with outcomes that permit insights into interplant competition. PMID- 16219080 TI - Do competition and herbivory alter the internal nitrogen dynamics of birch saplings? AB - Deciduous trees recycle nitrogen within their tissues. The aim of this study was to test the hypothesis that reductions in plant growth, caused by competition and herbivory, reduce the sink strength for N during autumn nutrient withdrawal, and reduce the storage capacity and hence the amount of N remobilized in the following spring. We used (15)N-labelled fertilizer to quantify N uptake, leaf N withdrawal and remobilization. Betula pubescens saplings were grown with either Molinia caerulea or Calluna vulgaris, and subjected to simulated browsing damage. Competition reduced B. pubescens leaf N withdrawal and remobilization, with C. vulgaris having a greater effect than M. caerulea. However, simulated browsing had no significant effect on sapling N dynamics. The patterns of leaf N withdrawal and remobilization closely followed sapling dry mass. We conclude that the effect of competition on sapling mass reduces their N-storage capacity. This reduces sink strength for leaf N withdrawal and the source strength for remobilized N. The ability of saplings to compensate for browsing damage removed any potential effect of browsing on N dynamics. PMID- 16219081 TI - Translocation of 15N indicates nitrogen recycling in the mat-forming lichen Cladonia portentosa. AB - Nitrogen translocation was measured in Cladonia portentosa during 2 yr growth in Scottish heathland. Translocation was predicted to occur if N is resorbed from senescent basal tissue and recycled within the thallus. (15)N was introduced into either the lower (TU thalli) or upper (TD thalli) 25 mm of 50-mm-long thalli as (15)N-NH(4) (+), (15)N-NO(3) (-) or (15)N-glycine. Labelled thalli were placed within intact lichen cushions, either upright (TU) or inverted (TD). Vertical distribution of label was quantified immediately following labelling and after 1 and 2 yr. Independently of the form of introduced label, (15)N migrated upwards in TU thalli, with new growth being a strong sink. Sink regions for (15)N during year 1 (including new growth) became sources of (15)N translocated to new growth in year 2. Upward migration into inverted bases was minimal in TD thalli, but was again marked in new growth that developed from inverted apices. Relocation of N to regions of growth could facilitate internal N recycling, a process postulated to explain the ecological success of mat-forming lichens. PMID- 16219082 TI - Phosphorus nutrition-mediated effects of arbuscular mycorrhiza on leaf morphology and carbon allocation in perennial ryegrass. AB - The aim of this work was to disentangle phosphorus status-dependent and independent effects of arbuscular mycorrhizal fungus (AMF) on leaf morphology and carbon allocation in perennial ryegrass (Lolium perenne). To this end, we assessed the P-response function of morphological components in mycorrhizal and nonmycorrhizal plants of similar size. AMF (Glomus hoi) stimulated relative P uptake rate, decreased leaf mass per area (LMA), and increased shoot mass ratio at low P supply. Lower LMA was caused by both decreased tissue density and thickness. Variation in tissue density was almost entirely caused by variations in soluble C, while that in thickness involved structural changes. All effects of AMF were indistinguishable from those mediated by increases in relative P-uptake rate through higher P-supply rates. Thus the relationships between relative P uptake rate, leaf morphology and C allocation were identical in mycorrhizal and nonmycorrhizal plants. No evidence was found for AMF effects not mediated by changes in plant P status. PMID- 16219084 TI - Loci controlling partial resistance to rice blast do not show marked QTL x environment interaction when plant nitrogen status alters disease severity. AB - Plant disease susceptibility is often increased by nitrogen (N) application. Therefore, it is important to know if resistance loci are effective in different plant N environments. One-hundred lines of the Bala x Azucena rice (Oryza sativa) mapping population were grown in two N treatments and tested for partial resistance to blast (Magnaporthe grisea) isolate CD100. Disease severity (DS), the number and size of lesions and plant N and C concentrations were measured and the results subject to quantitative trait loci (QTL) and QTL x environment analysis. There was a 66% higher plant N concentration in the high N treatment and DS increased significantly, mostly as a result of increased numbers of lesions. Nine regions contained QTL for disease traits but only one showed evidence of statistically significant QTL x treatment interaction. This was a large effect quantitative trait locus at marker R1933 on chromosome 12 which was less effective at high N. Apparently, blast disease is increased by higher plant N, but the efficacy of partial resistance genes is not greatly affected by N application. PMID- 16219083 TI - Physiological and molecular evidence for Pi uptake via the symbiotic pathway in a reduced mycorrhizal colonization mutant in tomato associated with a compatible fungus. AB - A Lycopersicon esculentum mutant (rmc) is resistant to colonization by most arbuscular mycorrhizal fungi (AMF), but one Glomus intraradices isolate (WFVAM 23) develops arbuscules and vesicles in the rmc cortex. It is unknown whether the symbiotic phosphate (Pi)-uptake pathway is operational in this interaction. Hyphal uptake of (32)Pi and expression of plant Pi transporter genes were investigated in the rmc mutant and its wild-type progenitor (76R) associated with three AMF. Hyphae transferred (32)Pi in all symbioses with 76R and in the rmc-G. intraradices WFVAM 23 symbiosis. The other AMF did not colonize rmc. The Pi transporter-encoding LePT1 and LePT2 were expressed constitutively or in P starved roots, respectively. The mycorrhiza-inducible Pi transporters LePT3 and LePT4 were expressed only in plants with AMF colonization and symbiotic (32)Pi transfer. LePT3 and LePT4 transcripts were reliable markers for a functional mycorrhizal uptake pathway in rmc. Our novel approach to the physiology and molecular biology of P transport can be applied to other arbuscular-mycorrhizal symbioses, irrespective of the size of plant responses. PMID- 16219086 TI - Modes and rates of selfing and associated inbreeding depression in the self incompatible plant Senecio squalidus (Asteraceae): a successful colonizing species in the British Isles. AB - The strength of the self-incompatibility (SI) response in Senecio squalidus was measured across its British range. Geographic variation in SI was investigated and the extent and inheritance of pseudo-self-compatibility (PSC) and inbreeding depression were determined. Mean self-fruit-set per capitulum was calculated for individuals and sample populations. The heritability of PSC and the magnitude of inbreeding depression were assessed by comparing selfing rates and fitness trait values between SI and PSC parent-progeny lines. SI was found to be strongly expressed in S. squalidus throughout its British range, with only 3.1% of the individuals sampled showing PSC. This PSC had relatively low heritability with stronger expression of SI in selfed progeny relative to PSC parents. Inbreeding depression was shown to be great in S. squalidus, with mean life history stage values ranging from 0.18 to 0.25. The strength of SI in S. squalidus appears not to have weakened in response to its rapid colonization of Britain. The avoidance of inbreeding depression is likely to be the primary factor maintaining strong SI in this successful colonizing species. PMID- 16219085 TI - A case study from the interaction of strawberry and Botrytis cinerea highlights the benefits of comonitoring both partners at genomic and mRNA level. AB - Strawberry Fragaria x ananassa (cv. Korona) was inoculated with Botrytis cinerea by dipping berries in a conidial suspension. Colonization by the pathogen was monitored using real-time PCR, ELISA and ergosterol assays, the first showing the highest sensitivity. The expression of pathogen beta-tubulin and six polygalacturonases (Bcpg1-6) and three host defence genes (polygalacturonase inhibiting protein (FaPGIP) and two class II chitinases) were monitored using real-time RT-PCR. The maximum transcript levels of the host defence genes occurred at 16 h postinoculation (hpi) at the presumed initial penetration stage. The unique transcript profile of Bcpg2 over the 96-h incubation time and its high transcript levels relative to those of the other Bcpgs at 8-24 hpi suggest that the gene has a specific role in the penetration stage. Bcpg1 was expressed constitutively at a relatively high level in actively growing mycelia throughout the experimental period. Comparison of the transcript profiles indicated that Bcpg1 and Bcpg3-6 were coordinately regulated. PMID- 16219087 TI - Evolution and maintenance of pollen-colour dimorphisms in Nigella degenii: habitat-correlated variation and morph-by-environment interactions. AB - Dimorphism in pollen colour is rare among flowering plants, but occurs in two geographically and morphologically distinct subspecies of Nigella degenii (Ranunculaceae). We evaluated the role of genotype-by-environment interactions in the maintenance of two pollen morphs within each of these subspecies. Morph frequencies in a number of populations were related to current habitat conditions, and an extensive common-garden experiment involving both optimal and stressful conditions (drought and nutrient deficiency) was carried out. The putatively derived (dark) pollen morph of N. degenii ssp. barbro has a higher frequency on slopes facing north or east than on slopes facing south or west. Plants of the dark morph also have a higher mortality under drought stress or nutrient deficiency. Data available for N. degenii ssp. jenny provide little evidence for habitat-correlated variation in morph frequency or morph-specific differences in fitness under optimal and stressful growth conditions. Our results suggest that morph-by-environment interactions in mortality could contribute to the maintenance of pollen-colour dimorphisms in N. degenii ssp. barbro. PMID- 16219088 TI - Diagnostic procedures in tularaemia with special focus on molecular and immunological techniques. AB - Tularaemia is a severe bacterial zoonosis caused by the highly infectious agent Francisella tularensis. It is endemic in countries of the northern hemisphere ranging from North America to Europe, Asia and Japan. Very recently, Francisella like strains causing disease in humans were described from tropical northern Australia. In the last decade, efforts have been made to develop sensitive and specific immunological and molecular techniques for the laboratory diagnosis of tularaemia and also for the definite identification of members of the species F. tularensis and its four subspecies. Screening for the keyword 'Francisella' a Medline search over the last decade was performed and articles describing diagnostic methods for tularaemia and its causative agent were selected. Besides classical microbiological techniques (cultivation, biochemical profiling, susceptibility testing) several new immunological and molecular approaches to identify F. tularensis have been introduced employing highly specific antibodies and various polymerase chain reaction (PCR)-based methods. Whereas direct antigen detection by enzyme-linked immunosorbent assay (ELISA) or immunofluorescence might allow early presumptive diagnosis of tularaemia, these methods--like all PCR techniques--still await further evaluation. Therefore, diagnosis of tularaemia still relies mainly on the demonstration of specific antibodies in the host. ELISA and immunoblot methods started to replace the standard tube or micro agglutination assays. However, the diagnostic value of antibody detection in the very early clinical phase of tularaemia is limited. Francisella tularensis is regarded as a 'highest priority' biological agent (category 'A' according to the CDC, Atlanta, GA, USA), thus rapid and reliable diagnosis of tularaemia is required not only for a timely onset of therapy, the handling of outbreak investigations but also for the surveillance of endemic foci. Only very recently, evaluated test kits for serological diagnosis of human tularaemia became available, while the introduction of standardized molecular techniques for detection and typing is still missing. PMID- 16219089 TI - Comparison of BSE prevalence estimates from EU countries for the period July to December 2001 to the OIE and EU GBR classifications. AB - Consequent upon the bovine spongiform encephalopathy (BSE) crisis, the European Union (EU) Commission enacted various decisions, which demanded that all bovine animals over 30 months of age should be examined by one of the approved rapid tests when slaughtered for human consumption. All cattle over 24 months of age subject to 'special emergency slaughtering' or died on the farm or in transit or suspect of BSE infection should also be examined by one of the approved rapid tests. According to a specific commission decision, Sweden and Finland were to test only a sample of bovine animals over 30 months of age subject to normal slaughter. Testing commenced on 1 January 2001. The authors evaluate the results of more than 5 million tests performed in the second semester 2001 from across the EU. The prevalence of BSE in the risk categories considered (emergency slaughter, fallen stock and healthy slaughtered), and the probability distribution of true-positive, false-positive and false-negative results are estimated by second-order Bayesian analysis. The results of the validation of tests performed in the EU are also considered by estimation of the probability distribution of their sensitivity and specificity. The prevalence of infection estimated in the cattle population of each EU country is compared against the criteria given in the OIE Terrestrial Animal Health Code and is also used to evaluate the consistency of the results of EU Geographical BSE Risk with the actual infection levels in the countries. Finally, the capability of the two current approaches to BSE surveillance (i.e. the testing of all slaughtered and dead cattle as applied in the EU and a surveillance system targeted at animals in risk categories only) to detect the infection in a given population are discussed. PMID- 16219090 TI - Detection of bovine viral diarrhoea virus infected cattle--testing tissue samples derived from ear tagging using an Erns capture ELISA. AB - A new diagnostic approach testing tissue samples derived from cattle ear tagging for bovine viral diarrhoea virus (BVDV) antigen in a commercially available antigen capture enzyme-linked immunosorbent assay (ACE) was developed. To validate this method, 99 positive and 469 negative samples were tested. With those samples the assay yielded a sensitivity of 100% and specificity of >or=99.6%. Serum and ear tissue samples from 11 persistently infected (PI) BVDV calves were tested. While serum samples were negative after intake of colostrum, the ear tissue samples could be detected positive for BVDV all the time. Testing multiple samples derived from the same ear from PI cattle yielded positive results and low variation. Using cattle ear tags combining the ear tag application with sampling of a small ear tissue plug and testing those tissue samples with an ACE could be a reliable and economic way of BVDV testing. PMID- 16219091 TI - Outbreak of clinical listeriosis in sheep: evaluation from possible contamination routes from feed to raw produce and humans. AB - We report the results of clinical and microbiological investigations on Listeria monocytogenes infections in a flock of 55 sheep and describe the implications for the safety of the raw milk and raw-milk cheeses produced in the on-farm dairy. The outbreak was caused by feeding grass silage, which was contaminated with 5 log10 CFU L. monocytogenes/g. Clinically, although having been fed from the same batch of silage, abortive (nine ewes), encephalitic (one ewe) and septicaemic (four ewes) forms of listeriosis were observed during the outbreak phase. As the starting point of feeding the contaminated silage was known we could calculate an incubation period of 18+/-2 and 26 days for the abortive and the encephalitic form of listeriosis, respectively. Pathologically, the septicaemic cases suffered from Listeria accumulation at comparable numbers in visceral organs but not in the brain. Only a single ewe developed central nervous symptoms and a rhomb encephalitis was immunohistologically confirmed. In this case the infection proceeded from the nasal mucosa into the brain, with no infections of the liver, spleen and other visceral organs. Sampling of the cheese production chain, the farm environment and the persons living at the farm revealed the exposure of a farm-worker to an isolate genetically indistinguishable from the outbreak clone, obviously through the consumption of faecally contaminated bovine raw milk. The cheese under processing was free of Listeria because, as a result of intensive consultations, the farmer ensured a proper acidification of the cheese. The epidemiological findings suggest that food safety matters should be assessed in any case where infection of food-producing animals with potential human pathogens is observed. PMID- 16219092 TI - Reliability evaluation of sampling plan fixed by Council Directive 91/68/EEC for the maintenance of officially brucellosis-free flock status. AB - The European Union (EU) strategy with respect to sheep and goat brucellosis aims to eradicate the infection and achieve officially brucellosis-free (OBF) status in all EU holdings and territories. Council Directive 91/68/EEC of 28 January 1991 states that to maintain OBF status of ovine or caprine holdings located outside an OBF territory, only a representative number of animals need to be tested annually. However, depending on the number of animals in a holding, this testing method risks non-detection of the infection, thereby reducing the efficacy of the brucellosis control plan. The recommended sampling procedure has a low sensitivity for detecting infection in medium-sized flocks; furthermore, the risk of not detecting re-infection in OBF flocks, particularly in territories that have not yet gained OBF status, is also not acceptable. Moreover, in large sized flocks, the Directive sampling procedure entails taking an excessive number of samples, which can be very expensive. The authors evaluated, by using statistical analyses and a simulation model based on field data, the possible consequences of the current EU strategy. It is suggested that the sampling criteria for the maintenance of OBF status in the EU should be modified and that a statistically based sampling method should be applied instead of the fixed percentage method that is currently in use. PMID- 16219093 TI - Recognition of a B-cell epitope of the VapA protein of Rhodococcus equi in newborn and experimentally infected foals. AB - The aim of this study was to evaluate the usefulness of the previously identified B-cell epitope TSLNLQKDEPNGRASDTAGQ of the VapA protein of Rhodococcus equi and its association with R. equi pneumonia. A modified peptide designated PN11-14 corresponding to the epitope was recognized by all sera from experimentally infected foals with virulent R. equi ATCC103+ containing the virulence plasmid but not by its plasmid-cured derivative ATCC103- strain. Marked levels of VapA specific immunoglobulin (Ig)G were detected in all sera from the ATCC103+ infected foals at 2 weeks after the infection. One control animal had high titres as determined by the peptide enzyme-linked immunosorbent assay (ELISA), indicating the ELISA may not absolutely differentiate between foals with R. equi pneumonia and healthy exposed foals in farms where the prevalence of disease is high. However, numbers of animals used were small. Further evaluation of the peptide ELISA with field samples is necessary to determine whether the assay is diagnostically useful. This study showed that levels of passive transfer of maternal IgG antibodies to the epitope in newborn foals could be measured. Interestingly, the maternally derived antibodies were found to significantly (P<0.05 by Student's t-test) decline 2 weeks after birth. Seroconversion against naturally occurring VapA expressing R. equi could be detected in some foals at 4 weeks of age. Antibodies to the epitope peaked and were significantly (P<0.05) greater in foals aged between 6 and 8 weeks. These results indicated that the peptide ELISA could be used to monitor anti-VapA antibodies in foals, particularly those at the age of 4-6 weeks. It is possible that the ELISA may be of some use as a diagnostic test on farms where R. equi is non-endemic. Further studies using large number of field samples are needed to verify this assumption. PMID- 16219094 TI - Protection against neonatal Escherichia coli diarrhoea in pigs by vaccination of sows with a new vaccine that contains purified enterotoxic E. coli virulence factors F4ac, F4ab, F5 and F6 fimbrial antigens and heat-labile E. coli enterotoxin (LT) toxoid. AB - The efficacy of a new vaccine against neonatal Escherichia coli diarrhoea in piglets containing purified F4ab, F4ac, F5 and F6 fimbriae and detoxified heat labile toxin (LT) was tested in challenge experiments by the method described by the European Pharmacopoeia (3rd edn, EDQM, Council of Europe, Strasbourg, France). A group of 11 young sows from a herd without E. coli problems was vaccinated 6-8 and 2-4 weeks prior to expected farrowing and another group of nine young sows were non-vaccinated controls. Escherichia coli antibody titres were determined in serum samples taken from the sows before first vaccination and before farrowing and in colostrum samples. The newborn piglets were allowed to suckle colostrum from their mother immediately after birth. The piglets were marked with individually numbered ear tags. Approximately 12 h after birth, 118 piglets from vaccinated sows and 79 piglets from non-vaccinated control sows were challenged by oral instillation of 5 ml of a freshly prepared culture of one of the challenge strains [O8:K87:F4ab (LT+) or O149:K91:F4ac (LT+) or O9:K30:F5 or O9:K103:F6 respectively]. The challenge cultures contained as a mean 6.8x10(9) CFU/ml. After challenge the piglets were observed for 7 days and mortality and morbidity were recorded. Vaccinated sows developed significant levels of antibody titres in colostrum and serum. Control sows stayed at a low/seronegative level. The protective efficacy was excellent because 66.7-87.5% of the piglets from vaccinated sows remained without clinical signs after challenge. Only 0.0-28.0% of the piglets from non-vaccinated sows remained healthy and more than 47.1% of the piglets in this group died after challenge. It is concluded that the new vaccine is very effective in protection of piglets against neonatal E. coli diarrhoea. PMID- 16219095 TI - Prevalence and associations of symptoms of upper extremities, repetitive strain injuries (RSI) and 'RSI-like condition'. A cross sectional study of bank workers in Northeast Brazil. AB - BACKGROUND: The repetitive strain injury syndrome (RSI) is a worldwide occupational health problem affecting all types of economic activities. We investigated the prevalence and some risk factors for RSI and related conditions, namely 'symptoms of upper limbs' and 'RSI-like condition'. METHODS: We conducted a cross-sectional study with 395 bank workers in Recife, Northeast Brazil. Symptoms of upper limbs and 'RSI-like condition' were assessed by a simple questionnaire, which was used to screen probable cases of RSI. The diagnosis of RSI was confirmed by clinical examination. The associations of potential risk factors and the outcomes were assessed by multiple logistic regression analysis. RESULTS: We found prevalence rates of 56% for symptoms of the upper limbs and 30% for 'RSI-like condition'. The estimated prevalence of clinically confirmed cases of RSI was 22%. Female sex and occupation (as cashier or clerk) increased the risk of all conditions, but the associations were stronger for cases of RSI than for less specific diagnoses of 'RSI-like condition' and symptoms of upper limbs. Age was inversely related to the risk of symptoms of upper limbs but not to 'RSI like' or RSI. CONCLUSION: The variation in the magnitude of risk according to the outcome assessed suggests that previous studies using different definitions may not be immediately comparable. We propose the use of a simple instrument to screen cases of RSI in population based studies, which still needs to be validated in other populations. The high prevalence of RSI and related conditions in this population suggests the need for urgent interventions to tackle the problem, which could be directed to individuals at higher risk and to changes in the work organization and environment of the general population. PMID- 16219096 TI - The effects of injection of bovine vaccine into a human digit: a case report. AB - BACKGROUND: The incidence of needlestick injuries in farmers and veterinary surgeons is significant and the consequences of such an injection can be serious. CASE PRESENTATION: We report accidental injection of bovine vaccine into the base of the little finger. This resulted in increased pressure in the flexor sheath causing signs and symptoms of ischemia. Amputation of the digit was required despite repeated surgical debridement and decompression. CONCLUSION: There have been previous reports of injection of oil-based vaccines into the human hand resulting in granulomatous inflammation or sterile abscess and causing morbidity and tissue loss. Self-injection with veterinary vaccines is an occupational hazard for farmers and veterinary surgeons. Injection of vaccine into a closed compartment such as the human finger can have serious sequelae including loss of the injected digit. These injuries are not to be underestimated. Early debridement and irrigation of the injected area with decompression is likely to give the best outcome. Frequent review is necessary after the first procedure because repeat operations may be required. PMID- 16219098 TI - A coarse-to-fine approach to prostate boundary segmentation in ultrasound images. AB - BACKGROUND: In this paper a novel method for prostate segmentation in transrectal ultrasound images is presented. METHODS: A segmentation procedure consisting of four main stages is proposed. In the first stage, a locally adaptive contrast enhancement method is used to generate a well-contrasted image. In the second stage, this enhanced image is thresholded to extract an area containing the prostate (or large portions of it). Morphological operators are then applied to obtain a point inside of this area. Afterwards, a Kalman estimator is employed to distinguish the boundary from irrelevant parts (usually caused by shadow) and generate a coarsely segmented version of the prostate. In the third stage, dilation and erosion operators are applied to extract outer and inner boundaries from the coarsely estimated version. Consequently, fuzzy membership functions describing regional and gray-level information are employed to selectively enhance the contrast within the prostate region. In the last stage, the prostate boundary is extracted using strong edges obtained from selectively enhanced image and information from the vicinity of the coarse estimation. RESULTS: A total average similarity of 98.76%(+/- 0.68) with gold standards was achieved. CONCLUSION: The proposed approach represents a robust and accurate approach to prostate segmentation. PMID- 16219099 TI - Does a pre-hospital emergency pathway improve early diagnosis and referral in suspected stroke patients?--Study protocol of a cluster randomised trial [ISRCTN41456865]. AB - BACKGROUND: Early interventions proved to be able to improve prognosis in acute stroke patients. Prompt identification of symptoms, organised timely and efficient transportation towards appropriate facilities, become essential part of effective treatment. The implementation of an evidence based pre-hospital stroke care pathway may be a method for achieving the organizational standards required to grant appropriate care. We performed a systematic search for studies evaluating the effect of pre-hospital and emergency interventions for suspected stroke patients and we found that there seems to be only a few studies on the emergency field and none about implementation of clinical pathways. We will test the hypothesis that the adoption of emergency clinical pathway improves early diagnosis and referral in suspected stroke patients. We designed a cluster randomised controlled trial (C-RCT), the most powerful study design to assess the impact of complex interventions. The study was registered in the Current Controlled Trials Register: ISRCTN41456865--implementation of pre-hospital emergency pathway for stroke--a cluster randomised trial. METHODS/DESIGN: Two-arm cluster-randomised trial (C-RCT). 16 emergency services and 14 emergency rooms were randomised either to arm 1 (comprising a training module and administration of the guideline), or to arm 2 (no intervention, current practice). Arm 1 participants (152 physicians, 280 nurses, 50 drivers) attended an interactive two sessions course with continuous medical education CME credits on the contents of the clinical pathway. We estimated that around 750 patients will be met by the services in the 6 months of observation. This duration allows recruiting a sample of patients sufficient to observe a 30% improvement in the proportion of appropriate diagnoses. Data collection will be performed using current information systems. Process outcomes will be measured at the cluster level six months after the intervention. We will assess the guideline recommendations for emergency and pre-hospital stroke management relative to: 1) promptness of interventions for hyperacute ischaemic stroke; 2) promptness of interventions for hyperacute haemorrhagic stroke 3) appropriate diagnosis. Outcomes will be expressed as proportions of patients with a positive CT for ischaemic stroke and symptoms onset < or = 6 hour admitted to the stroke unit. DISCUSSION: The fields in which this trial will play are usually neglected by randomised controlled trial (RCT). We have chosen the cluster-randomised controlled trial (C-RCT) to address the issues of contamination, adherence to real practice, and community dimension of the intervention, with a complex definition of clusters and an extensive use of routine data to collect the outcomes. PMID- 16219100 TI - The clopidogrel after surgery for coronary artery disease (CASCADE) randomized controlled trial: clopidogrel and aspirin versus aspirin alone after coronary bypass surgery [NCT00228423]. AB - BACKGROUND: Saphenous vein graft disease remains a major limitation of coronary artery bypass graft surgery. The process of saphenous vein intimal hyperplasia begins just days after surgical revascularization, setting the stage for graft atherosclerotic disease and its sequalae. Clopidogrel improves outcomes in patients with atherosclerotic disease, and is effective at reducing intimal hyperplasia in animal models of thrombosis. Therefore, the goal of this study will be to evaluate the efficacy of clopidogrel and aspirin therapy versus aspirin alone in the prevention of saphenous vein graft intimal hyperplasia following coronary artery bypass surgery. METHODS: Patients undergoing multi vessel coronary artery bypass grafting and in whom at least two saphenous vein grafts will be used are eligible for the study. Patients will be randomized to receive daily clopidogrel 75 mg or placebo, in addition to daily aspirin 162 mg, for a one year duration starting on the day of surgery (as soon as postoperative bleeding has been excluded). At the end of one year, all patients will undergo coronary angiography and intravascular ultrasound assessment of one saphenous vein graft as selected by randomization. The trial will be powered to test the hypothesis that clopidogrel and aspirin will reduce vein graft intimal hyperplasia by 20% compared to aspirin alone at one year following bypass surgery. DISCUSSION: This trial is the first prospective human study that will address the question of whether clopidogrel therapy improves outcomes and reduces saphenous vein graft intimal hyperplasia following cardiac surgery. Should the combination of clopidogrel and aspirin reduce the process of vein graft intimal hyperplasia, the results of this study will help redefine modern antiplatelet management of coronary artery bypass patients. PMID- 16219101 TI - Human immunodeficiency virus type-1 episomal cDNA in semen. AB - BACKGROUND: Episomal 2-long terminal repeat (LTR) HIV-1 cDNA, a by-product of HIV 1 infection, is used in clinical trials as a marker for ongoing viral replication. It would be useful to employ 2-LTR cDNA to monitor cryptic HIV-1 infection in the genital tract of men on antiretroviral therapy (ART) to predict the evolution of sexually transmissible drug-resistant HIV-1, but studies thus far have failed to detect this marker in semen. The objectives of this study were: 1) to use a technique that maximizes DNA recovery from HIV-1 infected white blood cells in semen to determine if episomal 2-LTR cDNA is detectable in semen of ART-naive men with other evidence of genital tract HIV-1 infection, and 2) to compare levels of HIV-1 2-LTR cDNA, RNA, and proviral DNA in semen from HIV-1+ men on ART. RESULTS: Using a somatic cell DNA extraction technique, 2-LTR cDNA was detected by PCR/ELISA in 4 out of 8 semen samples from ART-naive men selected for other signs of seminal HIV-1 infection (positive controls). Southern blot and DNA sequencing confirmed that the amplified sequences were HIV-1 2-LTR cDNA; copy numbers ranged from 55 to 504 copies/sample. Two semen samples from a cohort of 22 HIV-1-infected men on dual nucleoside therapy, one with and one without detectable seminal HIV-1 RNA, were 2-LTR cDNA positive (336 and 8,560 copies/sample). Following addition of indinavir to the therapy regimen, no semen samples from 21 men with controlled peripheral and seminal viral loads were 2-LTR cDNA positive at 1 and 6 month time points, despite the persistence of HIV-1 proviral DNA+ semen cells and seminal cytomegalovirus (CMV) shedding in some cases. However, one individual who failed indinavir therapy and later developed distinct protease inhibitor (PI) drug resistance mutations in semen, maintained elevated levels of HIV-1 RNA and 2-LTR cDNA in semen. CONCLUSION: 2-LTR HIV-1 cDNA is detectable in semen of HIV-1-infected men. Two men on ART had 2-LTR HIV-1 cDNA in semen, suggesting that this marker may prove to be useful to monitor HIV 1 infection in the genital tract of men on ART to predict the evolution of drug resistance mutations in semen. PMID- 16219102 TI - Medical record linkage in health information systems by approximate string matching and clustering. AB - BACKGROUND: Multiplication of data sources within heterogeneous healthcare information systems always results in redundant information, split among multiple databases. Our objective is to detect exact and approximate duplicates within identity records, in order to attain a better quality of information and to permit cross-linkage among stand-alone and clustered databases. Furthermore, we need to assist human decision making, by computing a value reflecting identity proximity. METHODS: The proposed method is in three steps. The first step is to standardise and to index elementary identity fields, using blocking variables, in order to speed up information analysis. The second is to match similar pair records, relying on a global similarity value taken from the Porter-Jaro-Winkler algorithm. And the third is to create clusters of coherent related records, using graph drawing, agglomerative clustering methods and partitioning methods. RESULTS: The batch analysis of 300,000 "supposedly" distinct identities isolates 240,000 true unique records, 24,000 duplicates (clusters composed of 2 records) and 3,000 clusters whose size is greater than or equal to 3 records. CONCLUSION: Duplicate-free databases, used in conjunction with relevant indexes and similarity values, allow immediate (i.e. real-time) proximity detection when inserting a new identity. PMID- 16219103 TI - Elevated levels of procoagulant microparticles in a patient with myocardial infarction, antiphospholipid antibodies and multifocal cardiac thrombosis. AB - Circulating procoagulant microparticles (MP) are pathogenic markers of enhanced coagulability associated to a variety of disorders and released from stimulated vascular cells. When derived from endothelial cells, MP were found characteristic of thrombotic propensity in primary antiphospholipid syndrome (APS). The prothrombotic status of a patient with antiphospholipid antibodies (APL), a past history of mesenteric vein thrombosis and presenting myocardial infarction and extensive intracardiac thrombosis was examined by measurement of circulating procoagulant MP. MP of platelet and endothelial origins were highly elevated with respect to values detectable in patients with myocardial infarction and no history of APS (6- and 3-fold elevation, respectively) or in healthy volunteers (13- and 25-fold elevation, respectively). In this particular patient, with moderate APL titer, a drastic release of procoagulant MP could have contributed to thrombus growth and the development of extensive intracardiac thrombosis. PMID- 16219104 TI - Assessing immunization data quality from routine reports in Mozambique. AB - BACKGROUND: Worldwide immunization coverage shows an increase in the past years but the validity of the official reports for measuring change over time has been questioned. Facing this problem, donor supported initiatives like the Global Alliance for Vaccine and Immunizations, have been putting a lot of effort into assessing the quality of data used, since accurate immunization information is essential for the Expanded Program on Immunization managers to track and improve program performance. The present article, discusses the practices on record keeping, reporting and the support mechanism to ensure data quality in Mozambique. METHODS: A process evaluation study was carried out in Mozambique in one district (Cuamba) in Niassa Province, between January and March 2003. The study was based on semi-structured interviews, participant observation and review of the data collection materials. RESULTS: Differences were found for all vaccine types when comparing facility reports with the tally sheets. The same applies when comparing facility reports with district reports. The study also showed that a routine practice during supervision visits was data quality assessment for the outpatient services but none related to data consistency between the tally sheets and the facility report. For the Expanded Program on Immunization, supervisors concentrated more on the consistency checks between data in the facility reports and the number of vaccines received during the same period. Meetings were based on criticism, for example, why health workers did not reach the target. Nothing in terms of data quality was addressed nor validation rules. CONCLUSION: In this paper we have argued that the quality of data, and consequently of the information system, must be seen in a broader perspective not focusing only on technicalities (data collection tools and the reporting system) but also on support mechanisms. Implications of a poor data quality system will be reflected in the efficiency of health services facing increased demands, with stagnant or decreasing resources. PMID- 16219105 TI - Randomized phase II--study evaluating EGFR targeting therapy with cetuximab in combination with radiotherapy and chemotherapy for patients with locally advanced pancreatic cancer--PARC: study protocol [ISRCTN56652283]. AB - BACKGROUND: Pancreatic cancer is the fourth commonest cause of death from cancer in men and women. Advantages in surgical techniques, radiation therapy techniques, chemotherapeutic regimes, and different combined-modality approaches have yielded only a modest impact on the prognosis of patients with pancreatic cancer. Thus there is clearly a need for additional strategies. One approach involves using the identification of a number of molecular targets that may be responsible for the resistance of cancer cells to radiation or to other cytotoxic agents. As such, these molecular determinants may serve as targets for augmentation of the radiotherapy or chemotherapy response. Of these, the epidermal growth factor receptor (EGFR) has been a molecular target of considerable interest and investigation, and there has been a tremendous surge of interest in pursuing targeted therapy of cancers via inhibition of the EGFR. METHODS/DESIGN: The PARC study is designed as an open, controlled, prospective, randomized phase II trial. Patients in study arm A will be treated with chemoradiation using intensity modulated radiation therapy (IMRT) combined with gemcitabine and simultaneous cetuximab infusions. After chemoradiation the patients receive gemcitabine infusions weekly over 4 weeks. Patients in study arm B will be treated with chemoradiation using intensity modulated radiation therapy (IMRT) combined with gemcitabine and simultaneous cetuximab infusions. After chemoradiation the patients receive gemcitabine weekly over 4 weeks and cetuximab infusions over 12 weeks. A total of 66 patients with locally advanced adenocarcinoma of the pancreas will be enrolled. An interim analysis for patient safety reasons will be done one year after start of recruitment. Evaluation of the primary endpoint will be performed two years after the last patient's enrollment. DISCUSSION: The primary objective of this study is to evaluate the feasibility and the toxicity profile of trimodal therapy in pancreatic adenocarcinoma with chemoradiation therapy with gemcitabine and intensity modulated radiation therapy (IMRT) and EGFR-targeted therapy using cetuximab and to compare between two different methods of cetuximab treatment schedules (concomitant versus concomitant and sequential cetuximab treatment). Secondary objectives are to determine the role and the mechanism of cetuximab in patient's chemoradiation regimen, the response rate, the potential of this combined modality treatment to concert locally advanced lesions to potentially resectable lesions, the time to progression interval and the quality of life. PMID- 16219106 TI - The spectrum of thyroid dysfunction in an Australian hepatitis C population treated with combination Interferon-alpha2beta and Ribavirin. AB - BACKGROUND: The study aims to assess the pattern of thyroid response to combination Interferon-alpha2beta (IFN-alpha) and Ribavirin (RBV) anti-viral therapy in an Australian hepatitis C cohort. These include the prevalence of thyroid dysfunction (TD) including hyperthyroidism and hypothyroidism and their possible predictors, the common overall pattern of thyroid function tests whilst receiving therapy and TD outcomes, and the correlation with HCV status outcome. METHODS: A retrospective analysis of all medical records was performed to assess thyroid function in Hepatitis C Virus (HCV) patients who were treated at the Hunter Area hepatitis C treatment center between 1995 and March 2004. The centre is part of the John Hunter hospital, a major tertiary referral centre in New South Wales, Australia. RESULTS: There were 272 cases available for review. The prevalence of TD is 6.7 percent and is made up predominantly of females (80 percent). There were 3 (1.1 percent) cases of hyperthyroidism with 2 (67 percent) females. Thirteen out of fifteen (80 percent) cases of hypothyroidism were females with the overall prevalence of 5.5 percent. The majority of hypothyroid patients still required Thyroxine supplement at the end of follow up. CONCLUSION: Ninety three percent of HCV treated patients have intact thyroid function at the end of treatment. The predominant TD is hypothyroidism. The predominant pattern of thyrotoxicosis (TTX) is that of thyroiditis although the number is small. Graves' like disease was not observed. People with pre-existing thyroid auto antibodies should be closely monitored for thyroid dysfunction, particularly hypothyroidism. PMID- 16219107 TI - Respiratory epithelial cells require Toll-like receptor 4 for induction of human beta-defensin 2 by lipopolysaccharide. AB - BACKGROUND: The respiratory epithelium is a major portal of entry for pathogens and employs innate defense mechanisms to prevent colonization and infection. Induced expression of human beta-defensin 2 (HBD2) represents a direct response by the epithelium to potential infection. Here we provide evidence for the critical role of Toll-like receptor 4 (TLR4) in lipopolysaccharide (LPS)-induced HBD2 expression by human A549 epithelial cells. METHODS: Using RTPCR, fluorescence microscopy, ELISA and luciferase reporter gene assays we quantified interleukin-8, TLR4 and HBD2 expression in unstimulated or agonist-treated A549 and/or HEK293 cells. We also assessed the effect of over expressing wild type and/or mutant TLR4, MyD88 and/or Mal transgenes on LPS-induced HBD2 expression in these cells. RESULTS: We demonstrate that A549 cells express TLR4 on their surface and respond directly to Pseudomonas LPS with increased HBD2 gene and protein expression. These effects are blocked by a TLR4 neutralizing antibody or functionally inactive TLR4, MyD88 and/or Mal transgenes. We further implicate TLR4 in LPS-induced HBD2 production by demonstrating HBD2 expression in LPS non responsive HEK293 cells transfected with a TLR4 expression plasmid. CONCLUSION: This data defines an additional role for TLR4 in the host defense in the lung. PMID- 16219109 TI - Development and validation of a quality of life questionnaire for patients with colostomy or ileostomy. AB - BACKGROUND: Quality of life of stoma patients is increasingly being addressed in clinical trials. However, the instruments used in the majority of these studies have not been validated specifically for stoma patients. The aim of this paper is to describe the development and validation of a quality-of-life instrument, "Stoma-QOL", specifically for patients with colostomy or ileostomy. METHODS: Potential items were formulated in English on the basis of the results of a series of semi-structured interviews with 169 adult stoma patients. The process resulted in a preliminary 37-item version, which was translated into French, German, Spanish and Danish, and administered repeatedly to 182 patients with colostomy or ileostomy. A psychometric selection of items was performed through Rasch Analysis. The measurement properties of the final questionnaire version were subsequently tested. RESULTS: The 20 items in the final questionnaire covered four domains--sleep, sexual activity, relations to family and close friends, and social relations to other than family and close friends. These items were found to define a unidimensional variable according to Rasch specifications (Infit MNSQ < 1.3). Internal consistency reliability calculated as Cronbach's alpha was 0.92, i.e., highly reliable. Spearman's correlation coefficients of scores across times of administration was > 0.88 (p < 0.01), indicating a high test-retest reliability. Item calibrations by country calculated as ICC were 0.81 (0.67-0.91 95% CI), confirming cross-cultural comparability across the European countries included in the study. CONCLUSION: Given the adequacy of the metric properties of the Stoma-QOL suggested by the psychometric analyses, this study confirms the suitability of the instrument in clinical practice and in clinical research. PMID- 16219110 TI - BioSysBio: Bioinformatics and Systems Biology Conference. Edinburgh, UK. 14-15 July 2005. Abstracts. PMID- 16219108 TI - A trial of somatic gene targeting in vivo with an adenovirus vector. AB - BACKGROUND: Gene targeting in vivo provides a potentially powerful method for gene analysis and gene therapy. In order to sensitively detect and accurately measure designed sequence changes, we have used a transgenic mouse system, MutaMouse, which has been developed for detection of mutation in vivo. It carries bacteriophage lambda genome with lacZ+ gene, whose change to lacZ-negative allele is detected after in vitro packaging into bacteriophage particles. We have also demonstrated that gene transfer with a replication-defective adenovirus vector can achieve efficient and accurate gene targeting in vitro. METHODS: An 8 kb long DNA corresponding to the bacteriophage lambda transgene with one of two lacZ negative single-base-pair-substitution mutant allele was inserted into a replication-defective adenovirus vector. This recombinant adenovirus was injected to the transgenic mice via tail-vein. Twenty-four hours later, genomic DNA was extracted from the liver tissue and the lambda::lacZ were recovered by in vitro packaging. The lacZ-negative phage was detected as a plaque former on agar with phenyl-beta-D-galactoside. RESULTS: The mutant frequency of the lacZ-negative recombinant adenovirus injected mice was at the same level with the control mouse (approximately 1/10000). Our further restriction analysis did not detect any designed recombinant. CONCLUSION: The frequency of gene targeting in the mouse liver by these recombinant adenoviruses was shown to be less than 1/20000 in our assay. However, these results will aid the development of a sensitive, reliable and PCR-independent assay for gene targeting in vivo mediated by virus vectors and other means. PMID- 16219112 TI - The health service use and cost of eating disorders. AB - BACKGROUND: The economic burden and health service use of eating disorders have received little attention, although such data are necessary to estimate the implications of any changes in clinical practice for patient care and health care resource requirements. This systematic review reports the current international evidence on the resource use and cost of eating disorders. METHOD: Relevant literature (1980-2002) was identified from searches of electronic databases and expert contacts. RESULTS: Two cost-of-illness studies from the UK and Germany, one burden-of-disease study from Australia and 14 other publications with relevant data from the UK, USA, Austria, Denmark and The Netherlands could be identified. In the UK, the health care cost of anorexia nervosa was estimated to be 4.2 million UK pounds in 1990. In Germany, the health care cost was 65 million Euro for anorexia nervosa and 10 million Euro for bulimia nervosa during 1998. The Australian study reported the health care costs of eating disorders to be Aus $22 million for year 1993/1994. Other costing studies focused mostly on in patient care reporting highly variable estimates. There is a dearth of research on non-health care costs. CONCLUSIONS: The limited available evidence reflects a general under-detection and under-treatment of eating disorders. Although both cost-of-illness studies may significantly underestimate the costs of eating disorders because of important omitted cost items, other evidence suggests that the economic burden is likely to be substantial. Comprehensive data on the resource use of patients with eating disorders are urgently needed for better estimations, and to be able to determine cost-effective treatment options. PMID- 16219113 TI - Mood- and restraint-based antecedents to binge episodes in bulimia nervosa: possible influences of the serotonin system. AB - BACKGROUND: In bulimic syndromes, binge episodes are thought to be caused by dietary restraint and negative moods. However, as central serotonin (5 hydroxytryptamine: 5-HT) mechanisms regulate appetite and mood, the 5-HT system could be implicated in diet- and mood-based binge antecedents. METHOD: We used hand-held computers to obtain repeated "online" measurements of eating behaviors, moods, and self-concepts in 21 women with bulimic syndromes, and modeled 5-HT system activity with a measure of platelet [3H]paroxetine-binding density. RESULTS: Mood and self-concept ratings were found to be worse before binge episodes (than at other moments), and cognitive restraint was increased. After binges, mood and self-concept deteriorated further, and thoughts of dieting became more intense. Intriguingly, lower paroxetine-binding density predicted poorer mood and self-concept before a binge, larger post-binge decrements in mood and self-concept, and larger post-binge increases in dietary restraint. CONCLUSIONS: Paroxetine binding thus seemed to reflect processes that impacted upon mood-related antecedents to binge episodes, and consequences implicating mood and dietary restraint. PMID- 16219114 TI - An empirical study of the typology of bulimia nervosa and its spectrum variants. AB - BACKGROUND: There is an ongoing debate about the best way to classify eating disorders. This study examined potential subtypes of bulimia nervosa. METHOD: Latent class analysis (LCA) was used to identify subtypes of bulimic symptomatology, utilizing data from 234 respondents in a cohort of black and white young women ( n =2054). Participants were administered gated screening questions from the Eating Disorders Examination to determine DSM-IV diagnoses of eating disorders. RESULTS: A 3-class solution was judged best. Class 1, the "purger subtype" ( n =116), was characterized by vomiting, the use of fasting/diet pills, and relatively little bingeing. Class 2, the "binger subtype" ( n =97) comprised women who engaged in bingeing but minimal compensatory behaviors. Class 3, the "binge-purger subtype" ( n =21) had relatively high rates of all symptoms. Findings of differences between the three subtypes on validator variables and differences between the three subtypes compared to non-eating disorder groups suggest validity of the three bulimic subtypes identified in our analyses. Ethnicity and class membership were associated [chi 2 (3)=21.89, p <0.0001], reflecting a greater percentage of white women than black women in Class 1 and a greater percentage of black women than white women in Class 2. CONCLUSIONS: LCA revealed one subtype that was similar to bulimia nervosa and two subtypes of bulimic symptomatology that did not meet criteria for bulimia nervosa yet appear to be clinically significant. Further study of the psychological correlates, course, and treatment response of these classes would be of clinical interest. PMID- 16219115 TI - Age at onset and familial risk for major depression in a Swedish national twin sample. AB - BACKGROUND: In many biomedical disorders, early age at onset (AAO) is an index of high liability to illness which is manifest by an increased risk of illness in relatives. Most but not all prior studies report such a pattern for major depression (MD). METHOD: Lifetime MD and AAO were assessed at personal interview using modified DSM-III-R criteria in 13864 twin pairs, including 4229 onsets of MD, from the Swedish National Twin Registry. Analyses were conducted using Cox proportional hazards models. RESULTS: Controlling for year of birth, gender, zygosity, co-twin history of MD and the interaction of zygosity and co-twin history, the best-fit model showed a significant main effect and a quadratic effect of AAO of MD in the co-twin on the log hazard ratio for MD in the index twin. When examined together, these effects predicted that from the ages of 15 to approximately 35 years, AAO of MD is moderately negatively related to risk of illness in relatives. However, past age 35, the function flattens out, with little change of risk in relatives with further increases of AAO. Even when the co-twin had a late AAO, the risk in the index twin substantially exceeded that seen when the co-twin had no history of MD. CONCLUSION: In this large sample, AAO is a meaningful, albeit modest, index of familial liability to MD. The relationship is nonlinear and results largely from an increased liability in individuals with an early AAO. These results should be interpreted in the context of the limitations of long-term recall. PMID- 16219116 TI - Heritability of life satisfaction in adults: a twin-family study. AB - BACKGROUND: Subjective well-being (SWB) can be partitioned into the components life satisfaction and affect. Research on factors influencing these components of well-being has mainly focused on environmental characteristics. The aim of this study was to investigate the relative contribution of genes and environment to individual differences in life satisfaction in a large sample of Dutch twins and their singleton siblings. METHOD: Life satisfaction of 5668 subjects registered with The Netherlands Twin Registry (NTR) was measured with a Dutch version of the self-reported Satisfaction with Life Scale. An extended twin design was used to obtain correlations in life satisfaction scores for monozygotic twins, dizygotic twins and sibling pairs and to estimate the contribution of genes and environment to the variation in life satisfaction. RESULTS: No differences between males and females were found in the mean level of life satisfaction. Broad-sense heritability was 38%. Non-additive genetic factors explained all or most of the genetic influences. The remaining 62% of the variance in life satisfaction could be attributed to unique environmental factors, both persistent and transitory, plus measurement error. CONCLUSIONS: Individual differences in life satisfaction are determined in part by genetic factors that are largely or entirely non additive in nature. PMID- 16219117 TI - Neuregulin 1 gene and variations in perceptual aberration of schizotypal personality in adolescents. AB - BACKGROUND: We test the hypothesis that the neuregulin 1 (NRG1 ) gene at chromosome 8p22-p12, which has been implicated as a susceptibility gene to schizophrenia, is associated with variations in schizotypal personality in non clinical populations. METHOD: A randomly selected sample of 905 adolescents were assessed for their personality features using the Perceptual Aberration Scale (PAS) and the Schizotypal Personality Questionnaire (SPQ) and genotyped for three single nucleotide polymorphisms (SNP8NRG221533, rs3924999, and rs2954041) at the NRG1 gene. Relations between the three genetic variants and continuous schizotypal personality scores were evaluated using ANOVA for single-locus analyses and haplotype trend regression test for multi-locus analyses. RESULTS: Single locus analysis showed that the A allele of rs3924999, a functional polymorphism in exon 2, had the largest effect size and exhibited a prominent allele-dose trend effect for the PAS score. Haplotype analyses using the haplotype trend regression test indicated that the A allele of rs3924999 was mainly responsible for the association with the PAS but not with the SPQ or its three factors, and the magnitude of significance was not strengthened by the combination of this allele with adjacent locus. CONCLUSIONS: Our study provides the first evidence for the association of NRG1 with schizotypal personality and indicates a possible role of NRG1 in the genetic etiology of schizophrenia through perceptual aberrations. PMID- 16219118 TI - Neuregulin 1 (NRG1 ) and schizophrenia: analysis of a US family sample and the evidence in the balance. AB - BACKGROUND: Individual genome-wide linkage scans and meta-analyses support that one or more susceptibility genes for schizophrenia are located in chromosome 8p. A gene from this region, neuregulin 1 (NRG1 ), known to be involved with glutamatergic function, has been found to be associated in some studied samples. METHOD: We have examined a new combined schizophrenia sample with 136 schizophrenia families largely of European ancestry (EA) and 646 subjects with DNA. We genotyped 14 single nucleotide polymorphisms (SNPs) in NRG1 including those reported to comprise schizophrenia-associated haplotypes in Icelandic, Scottish, Irish, and Chinese Han populations. RESULTS: We found no evidence of association at a single-marker or a haplotypic level. We review methodological aspects of previous studies to enable us to put our findings into context. CONCLUSIONS: Our failure to find an association between NRG1 and schizophrenia might reflect different linkage disequilibrium (LD) patterns found in different populations, disease allelic heterogeneity, clinical heterogeneity of schizophrenia, or inadequate statistical power deriving from moderate sample size. NRG1, if a true gene for schizophrenia, accounts for a small fraction of the disease in most populations. The confirmation of NRG1 as a schizophrenia susceptibility gene will require studies with a comprehensive set of markers and in larger samples. The possibility remains that reports of NRG1 association might reflect false positives. PMID- 16219119 TI - A direct interview family study of obsessive-compulsive disorder. I. AB - BACKGROUND: This and the companion paper present two sequential family studies of obsessive-compulsive disorder (OCD) conducted by the same research group, but with different sampling and best-estimate procedures. In addition to providing further data on familial transmission of OCD, we used comparison of disparate findings (moderate, specific familial aggregation in this first study versus a stronger effect for other anxiety disorders than for OCD alone in the second) to examine possible effects of proband characteristics and informant data on outcome. METHOD: In this initial study we interviewed 179 first-degree relatives of 72 OCD probands and 112 relatives of 32 never mentally ill (NMI) controls. Informant data were obtained on an additional 126 relatives (total "combined" samples of 263 and 154 respectively). Analyses used best-estimate diagnoses made by consensus of two "blinded" senior clinicians who reviewed all diagnostic materials including proband informant data about relatives. RESULTS: Significantly higher risk for OCD but not other anxiety disorders was found in relatives of OCD probands compared to relatives of controls in both the directly interviewed and combined samples. There was no relationship between proband age at onset of OCD and strength of familial aggregation. CONCLUSIONS: These data indicate moderate familial aggregation of OCD, but do not support increased transmission by early onset probands, or a familial relationship between OCD and other anxiety disorders with the possible exception of generalized anxiety disorder. PMID- 16219120 TI - A direct interview family study of obsessive-compulsive disorder. II. Contribution of proband informant information. AB - BACKGROUND: Overall findings of our first direct interview family study of obsessive-compulsive disorder (OCD) indicated that OCD is familial. In this replication study, we carefully examined the role of informant data in ascertaining OCD in relatives. METHOD: We interviewed 112 relatives of 57 OCD patients and 115 relatives of 41 not ill controls predominantly by telephone. Additional analyses included a combined sample of relatives about whom any diagnostic information was available (228 OCD and 239 controls). To examine the contribution of proband information about relatives, we considered two sets of best-estimate diagnoses. First, we ascertained best-estimate diagnoses for relatives using information from direct interviews and from all informants except the proband. Then, we re-diagnosed relatives based on all available information, including reports from the proband about their relatives. RESULTS: When relative diagnoses were derived without the benefit of proband informant reports, no evidence of familial OCD transmission was found. When diagnoses were made including information from the proband about the relative, evidence of familial OCD was found, but only when the diagnostic threshold was lowered to include cases with probable OCD or OCD symptoms. Other diagnoses (generalized anxiety disorder, social phobia, drug use disorder) were also higher among OCD relatives. CONCLUSIONS: This second study provides less robust support for familial transmission of OCD. Evidence for familial transmission of OCD was found only when diagnoses were made using information from the affected proband about their relatives. Taken in context of past findings, our own inconsistent results suggest that OCD may be heterogeneous with regard to familial transmission. Also, more careful attention should be paid to the contribution of informant reports, especially from relatives affected by the same disorder. PMID- 16219122 TI - Does the addition of cognitive behavioral therapy improve panic disorder treatment outcome relative to medication alone in the primary-care setting? AB - BACKGROUND: Randomized clinical trials indicate a benefit from combining medications with cognitive behavioral therapy (CBT) relative to medication alone for panic disorder. Using an as-treated analysis, we evaluated whether the addition of CBT enhanced outcomes for panic disorder relative to medications alone in the primary-care setting. METHOD: Primary-care patients with panic disorder reported on their receipt of CBT and medications over the 3 months following baseline assessment. The degree to which outcomes for those who used anti-panic medications were enhanced by the receipt of at least one component of CBT was analyzed using a propensity score model that took into account observable baseline patient characteristics influencing both treatment selection and outcomes. RESULTS: The addition of CBT resulted in statistically and clinically significant improvements at 3 months on anxiety sensitivity, social avoidance, and disability. Also, patients receiving CBT in the first 3 months of the study were more improved at 12 months than patients who took medications only during the first 3 months of the study. CONCLUSIONS: The clinical utility of the findings are discussed in terms of the importance of primary-care physicians encouraging their panic disorder patients to receive CBT as well as medications. PMID- 16219121 TI - The clinical effectiveness of guided self-help versus waiting-list control in the management of anxiety and depression: a randomized controlled trial. AB - BACKGROUND: There are significant barriers to accessing effective psychological therapy in primary care resulting from a lack of suitably trained therapists to meet current demand. More efficient service delivery using minimal interventions (such as bibliotherapy) provided by paraprofessional therapists may be one method of overcoming these problems, and is the subject of attention in the UK and elsewhere. A randomized trial was conducted to test the clinical effectiveness of this model. Assistant psychologists delivered a guided self-help intervention to patients with anxiety and depression who were currently waiting for psychological therapy. METHOD: A total of 114 patients were randomized either to guided self help or a waiting-list control group. All patients were followed up 3 months later, prior to starting conventional psychological therapy. Measures included self-reported adherence to the intervention, anxiety and depressive symptoms, social functioning and patient satisfaction. RESULTS: Adherence to the guided self-help intervention was acceptable and patients reported satisfaction with the intervention. However, there were no statistically significant differences between groups in anxiety and depression symptoms at 3 months. CONCLUSIONS: The results demonstrate that this model of guided self-help did not provide additional benefit to patients on a waiting list for psychological therapy. The results are considered in the context of possible internal and external validity threats, and compared with previous trials of minimal interventions. The implications of the results for the design of future minimal interventions are considered. PMID- 16219123 TI - Listening to patients' needs to improve their subjective quality of life. AB - BACKGROUND: Subjective quality of life has gained a crucial role as a global measure of outcome in mental health care. This study aimed to investigate the impact of meeting needs for care, as assessed by both patients and mental health professionals, to improve the subjective quality of life in a sample of patients receiving community-based psychiatric care. METHOD: The study was conducted using a 4-year prospective longitudinal design. A cohort of patients from the South Verona Community-based Mental Health Service (CMHS) was assessed at baseline and follow-up using, among other social and clinical measures, the Camberwell Assessment of Need (both staff and patient versions) and the Lancashire Quality of Life Profile. Predictors of changes of subjective quality of life were explored using block-stratified multiple regression procedures. RESULTS: Improvement in patients' clinical conditions as well as the reduction in patient rated unmet needs in the social domain predicted an increase in subjective quality of life over 4 years; changes in staff-rated needs did not show any association with changes in subjective quality of life. CONCLUSIONS: Meeting self perceived social needs, beyond symptoms reduction, seems to be of particular importance for ensuring a better quality of life for people with mental disorders. If the main goal of mental health care is to improve the quality of life of users, a policy of actively addressing patient-rated needs should be implemented. PMID- 16219124 TI - Presentation and frequency of catatonia in new admissions to two acute psychiatric admission units in India and Wales. AB - BACKGROUND: There are no modern cross-cultural comparative studies of the frequency and clinical presentation of catatonia in a Western country and India using standardized rating instruments and diagnostic criteria. METHOD: A total of 104 consecutively admitted patients in Wales and in India were screened for catatonic features using the same standardized rating instrument by the same psychiatrist to generate DSM-IV and other diagnostic criteria for catatonia, and a profile of signs in catatonia. Inter-rater reliability for the ratings made by the research psychiatrist was established with local psychiatrists at each unit. RESULTS: The frequency of DSM-IV criteria catatonia was 13.5% in India versus 9.6% in Wales ( N . S .). The severity of catatonia did not differ between the two units. However, retarded catatonia was more common in India (12.5%) versus Wales ( p <0.05) whereas the frequency of excited catatonia was equally common in both units. Catatonia was found in many different mental disorders not just schizophrenia and affective disorder. CONCLUSIONS: Catatonia is commonly found among psychiatric in-patients with a similar frequency and severity but differing clinical presentations in Wales and India. Some classic signs of catatonia like posturing, catalepsy, staring and stupor were more frequent among psychiatric admissions in India than Wales. The differing clinical presentations may be due to differences in demographic features rather than cultural or aetiological factors. PMID- 16219127 TI - [Staging 915 cases of nasopharyngeal carcinoma after simple radical radiotherapy- checkout of Fuzhou staging system (1992)]. AB - BACKGROUND & OBJECTIVE: Along with the development of treatments, different tumor staging systems are coexisted and have been modified. This study was to validate the rationality of the Fuzhou staging system (1992) of nasopharyngeal carcinoma (NPC), and to provide some suggestions. METHODS: A total of 915 NPC patients received radical radiotherapy alone in Cancer Center of Sun Yat-sen University from Jan. 1997 to Dec. 1998. The 1-, 3-, and 5-year follow-up rates were 98.7%, 95.2%, and 91.7%, respectively. The survival data were analyzed with Life table, Cox regression, Kaplan-Meier, and log-rank methods. RESULTS: The 1-, 3-, and 5 year overall survival rates of the 915 patients were 87.69%, 72.73%, and 64.44%; the 1-, 3-, and 5-year disease-freely survival rates were 86.87%, 69.72%, and 58.33%, respectively. Cox regression analysis showed that the 5-year survival statuses of the 915 patients were significantly correlated with their age and the tumor stage classified by the Fuzhou staging system (1992); the 5-year survival statuses of the 803 patients no more than 60 years old were only significantly correlated with tumor stage, and had no correlation with their age. Life table analysis validated that the tumor stage classified by Fuzhou staging system (1992) can roughly predict the prognosis, but the difference between the 5-year survival rates of stage I and II patients was not significant. Kaplan-Meier analysis showed no significant difference between survival statuses of stage T1 and T2 patients when adjusted by N classification. Therefore, we adjusted stage T2 without parapharyngeal space invasion to stage T1, stage T3 with carotid vagina invasion to stage T2, stage T4 with paranasal sinus involvement to stage T3, stage T3 with cranial nerve injury to stage T4, and stage N1 with bilateral lymph nodes involvement to stage N2. After the modifications, the differences among stage I to IVa, stage T1 to T4 (adjusted by N stage), or stage N0 to N3 (adjusted by T stage) were significant. CONCLUSION: Taking the impact of age on the prognosis and the interaction between T stage and N stage into consideration, the above modifications of should be included when renewing the Fuzhou staging system (1992). PMID- 16219128 TI - [Effect of RNAi-mediated gene silencing of C-erbB-2 on proliferation of lung adenocarcinoma cell line calu-3]. AB - BACKGROUND & OBJECTIVE: C-erbB-2 gene is amplified or overexpressed in breast cancer, ovarian cancer, and lung cancer, and is related with enhanced malignancy and metastatic ability, intrinsic chemoresistance, and poor prognosis of tumors. RNA interfering (RNAi), a new genetic technique, can efficiently and specifically suppress gene expression. This study was to investigate the effect of small interfering RNA (siRNA)-mediated gene silencing of C-erbB-2 on proliferation of human lung adenocarcinoma cell line calu-3. METHODS: C-erbB-2 siRNA was transfected into calu-3 cells; cell morphology was observed under light microscope. The mRNA and protein levels of C-erbB-2 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM). The proliferation of calu-3 cells was assessed by MTT assay. Cell cycle and apoptosis were analyzed by FCM. RESULTS: C-erbB-2 siRNA down-regulated the mRNA and protein levels of C-erbB-2 in calu-3 cells; 48 h after transfection of C-erbB-2 siRNA, the protein level of C-erbB-2 was markedly decreased. The positive rate of C-erbB 2 was significantly lower in C-erbB-2 siRNA group than in untransfected group, empty vector group, and nonspecific siRNA group [(25.04+/-1.56)% vs. (98.24+/ 2.23)%, (95.67+/-1.98)%, and (94.79+/-0.87)%, P < 0.01]. C-erbB-2 siRNA inhibited proliferation of calu-3 cells: G(0)/G(1) phase proportion of C-erbB-2 siRNA group was significantly higher than that of untransfected group [(56.6+/-3.6)% vs. (45.5+/-3.2)%, P < 0.01]. C-erbB-2 siRNA also enhanced cell apoptosis. CONCLUSION: Specific siRNA targeting C-erbB-2 can effectively inhibit C-erbB-2 expression and proliferation of calu-3 cells. PMID- 16219129 TI - [A DNA vaccine encoding the extracellular domain of porcine endoglin induces antitumor immunity in a mouse colon carcinoma model]. AB - BACKGROUND & OBJECTIVE: Endoglin is a marker of tumor angiogenesis. Increasing evidences proved that passive immunotherapy with anti-endoglin monoclonal antibody can effectively inhibit tumor growth, and xenogeneic homologous DNA vaccine can inhibit cross antitumor immunity. This study was to explore the inhibitory effect of a DNA vaccine encoding the extracellular domain of porcine endoglin (ppEDG) on tumor growth in a mouse colon carcinoma model. METHODS: ppEDG was used as a DNA vaccine to actively immunize the colon carcinoma-bearing mice. Tumor volume and survival rate of the mice were observed in 3-day intervals. Microvessel density (MVD) was detected by immunohistochemistry; antibodies against self-endoglin were detected by Western blot and ELISA; the B cells that secrete auto-antibodies against self-endoglin were detected by ELISPOT assay. RESULTS: Eighteen days after tumor cell inoculation, the tumor volume was significantly smaller in ppEDG-immunized group than in empty plasmid (e-p) group and normal saline (NS) group (P<0.05), and the survival time was significantly longer in ppEDG-immunized group than in the control groups (P<0.001). MVD was significantly lower in ppEDG-immunized group than in e-p group and NS group (19.2+/-4.5 vs. 76.9+/-14.4 and 81.4+/-16.9, P<0.001). The antibodies against self-endoglin were identified in ppEDG-immunized group; the major subtypes were IgG(1) and IgG(2b). The auto-antibody-producing B cells were much more in ppEDG immunized group than in e-p group and NS group (82.5+/-14.1 vs. 3.6+/-1.3 and 4.7+/-2.0, P<0.001). CONCLUSION: ppEDG DNA vaccine could induce the production of auto-antibodies against self-endoglin, which further inhibit angiogenesis and growth of colon carcinoma. PMID- 16219131 TI - [Effect of NK4 gene transfection on tumor growth of human pancreatic cancer xenograft in nude mice]. AB - BACKGROUND & OBJECTIVE: NK4 is not only an antagonist of hepatocyte growth factor but also an angiogenesis inhibitor. Studies have confirmed that NK4 can inhibit tumor growth and metastasis, but its effect on pancreatic cancer remains unknown. This study was designed to observe the effect of NK4 gene on human pancreatic cancer in nude mice and the possible mechanisms. METHODS: The nude mouse model of pancreatic cancer was established with human pancreatic cancer cell line SW1990. The eukaryotic expression vector of NK4 gene was constructed and transfected into the tumors. The mice weight, tumor size and volume were measured before and after transfection. The apoptotic cells, microvessel density (MVD), and the expression of proliferating cell nuclear antigen (PCNA) in the tumors were observed using immunohistochemistry and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) technique. RESULTS: Four weeks after NK4 gene transfection, the tumor volume and weight was significantly smaller in NK4-transfected group than in PBS control group and empty vector group [(1.39+/-0.33) cm(3) vs. (2.06+/ 0.55) cm(3) and (1.90+/-0.36) cm(3), P<0.01; (1.30+/-0.81) g vs. (3.45+/-1.88) g and (3.14+/-1.51) g, P<0.01]; the inhibition rate was 62.29%. The tumor cell apoptotic index was significantly higher in NK4-transfected group than in the rest 2 groups (9.34+/-0.91 vs. 4.13+/-0.79 and 3.94+/-1.03, P<0.001); the MVD was significantly lower in NK4-transfected group than in the rest 2 groups (12.24+/ 4.63 vs. 20.13+/-7.00 and 19.70+/-6.15, P<0.05); the expression of PCNA in NK4 transfected group was not different from those of the rest 2 groups (53.88+/-4.30 vs. 56.24+/-4.03 and 54.33+/-5.41,P>0.05). CONCLUSION: NK4 gene transfection may inhibit the growth of human pancreatic cancer in mouse model through suppressing angiogenesis and enhancing the apoptosis of pancreatic cancer cells. PMID- 16219130 TI - [Cell differentiation during carcinogenesis of cervical epithelia]. AB - BACKGROUND & OBJECTIVE: The disturbance of cell differentiation is an important characteristic of tumor, but, up to date, the researches on this aspect are not as deep as those on cell proliferation of tumor. This study was to detect the alterations of cell differentiation and relative regulating factors during the carcinogenesis of cervical epithelia. METHODS: The protein expression of keratin (KT) 10, 14, 19 and beta1-integrin and beta-catenin in 20 specimens of normal squamous epithelium (NSE), 19 specimens of dysplasia squamous epithelial hyperplasia (DSEH), 39 specimens of squamous carcinoma in situ (SCIS), and 20 specimens of invasive squamous epithelial carcinoma (ISEC) were detected by inmmunohistochemistry; the mRNA expression of of beta1-integrin and beta-catenin were detected by in situ hybridization. RESULTS: The strong positive rates of KT10 in NSE, DSEH, SCIS, and ISEC were 85.0%, 52.6%, 18.0% and 0, respectively, with a descending trend; it was significantly lower in SCIS and ISEC than in NSE and DSEH (Chi(2)=11.28, P<0.05; Chi(2)=8.53, P<0.05). The expression of KT14 and KT19 showed a heterogeneity in SCIS and ISEC: the proteins were negative in some cases, but overexpressed in other cases with positive cells moved upwards from the basal layer; positive cells scattered at the full epithelial layer of SCIS. The expression of beta1-integrin also showed a descending trend in NSE, DSEH, SCIS, and ISEC. The positive rate of beta1-integrin was significantly lower in ISEC than in NSE and DSEH (Chi(2)=7.62, P <0.05; value of exact probability=0.014, P < 0.05); the mRNA expression of beta1-integrin showed the same trend as its protein expression in the samples although the difference were not significant. The protein and mRNA expression of beta-catenin was located in nuclei or cytoplasm of SCIS and ISEC; increased positive cells moved upwards from the basal layer. CONCLUSIONS: Restraint of terminal differentiation of epithelial cells may relate to the abnormal expression of beta1-integrin and the dysfunction of Wnt signaling pathway on regulating cell differentiation during the carcinogenesis of cervical epithelia. The expression of KT10 and beta1-integrin in SCIS is not obviously different from that in ISEC. PMID- 16219132 TI - [Enhancive effect of histone deacetylase inhibitor trichostatin a on transfection efficiency of adenovirus in ovarian carcinoma cell line A2780]. AB - BACKGROUND & OBJECTIVE: The presence of Coxsackie and adenovirus receptor (CAR) on target cell surface is required for efficient adenovirus transfection; lack or down-regulated expression of CAR on cancer cells is the main cause of inefficiency of adenovirus-based gene therapy. This study was to evaluate enhancive effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on the transfection efficiency of adenovirus in ovarian carcinoma cell line A2780, and explore its possible application to adenovirus-based gene therapy. METHODS: mRNA and protein levels of CAR on A2780 cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot before and after treatment of TSA. Transfection efficiency of adenovirus was valued by flow cytometry (FCM). In vitro antitumor effect of adenovirus/thymidine kinase (ADV/TK) was detected by MTT assay. RESULTS: After treatment of TSA, mRNA and protein levels of CAR on A2780 cells were obviously increased. Transfection rates of adenovirus were (1.24+/-0.14)% in untreated group, (7.58+/-0.32)% in 5 nmol/L of TSA treated group, and (7.94+/-0.28)% in 100 nmol/L of TSA treated groups. In vitro antitumor effect of ADV/TK was 4-10 folds in TSA (5 or 100 nmol/L) treated groups compared with that in untreated group. CONCLUSION: TSA could enhance transfection efficiency of adenovirus in ovarian carcinoma cells, and may be useful in gene therapy for ovarian carcinoma. PMID- 16219133 TI - [Inhibitory effect of 5-Aza-2'-deoxycytidine on human nasopharyngeal carcinoma xenograft in nude mice]. AB - BACKGROUND & OBJECTIVE: 5-Aza-2'-deoxycytidine (5-Aza-CdR) is an inhibitor of DNA methyltransferase; it may reactivate methylated antioncogene, therefore, inhibit the growth of cancer cells. This study was to observe the inhibitory effect of 5 Aza-CdR on the growth of human nasopharyngeal carcinoma (NPC) cells and xenografts in nude mice, explore the possible mechanisms, and search for new treatment target of NPC. METHODS: NPC cell line CNE cells were treated with 5-Aza CdR; the methylation status of death-associated protein kinase (DAPK) gene was evaluated by methylation-specific polymerase chain reaction (PCR). The model of human NPC xenograft in nude mice was constructed and treated with 5-Aza-CdR; the xenograft growth in nude mice was observed, and the mRNA and protein expression of DAPK was detected by reverse transcription-PCR (RT-PCR) and immunohistochemistry. RESULTS: No expression of DAPK mRNA was found in CNE cells and the xenografts in nude mice without treatment of 5-Aza-CdR. After treatment, the expression of DAPK mRNA in CNE cells and the xenografts was increased along with the increasing concentration of 5-Aza-CdR; the growth of CNE cells and the xenografts in nude mice were obviously inhibited, and the methylated DAPK gene was reactivated. Four weeks after treatment, no significant difference was found in body weight of nude mice between 5-Aza-CdR group and control group [(22.35+/ 2.02) g vs. (21.68+/-2.14) g, t=0.011, P>0.05]; the volume of xenografts was significantly smaller in 5-Aza-CdR group than in control group [(195.32+/-27.57) mm(3) vs. (343.67+/-23.08) mm(3), t=10.11, P<0.01]. CONCLUSION: 5-Aza-CdR may reactivate antioncogene silenced by de novo methylation, therefore, inhibit the growth of CNE cells in vivo and in vitro. PMID- 16219134 TI - [Correlation of GLC-82 lung carcinoma cell aggregation in suspension culture to the activation of protein kinases FAK, AKT, ERK, and SRC]. AB - BACKGROUND & OBJECTIVE: Our previous study revealed that tyrosine kinase FAK can partly mediate the aggregation of tumor cells in suspension culture, therefore, suppress cell apoptosis and promote cell proliferation. However, the downstream pathway of FAK in the formation of cell aggregation is unclear. This study was to investigate the roles of FAK and its potential downstream molecules ERK, AKT, and SRC in mediating aggregation of lung adenocarcinoma GLC-82 cells in suspension culture. METHODS: Morphology of the aggregations of GLC-82 cells and normal lung HBE cells in polyHEMA suspension culture was observed. Cell apoptosis was studied by DNA electrophoresis on agarose gel. The ability of cell transformation was studied by colony formation assay. The phosphorylation of the protein kinases was investigated by Western blot. FAK was silenced by RNA interference. RESULTS: GLC 82 cells survived and aggregated in suspension culture; while apoptosis occurred in HBE cells which formed no aggregation in suspension culture. Phosphorylation levels of FAK, ERK, AKT, and SRC were related to GLC-82 cell aggregation. FAK silencing partly blocked the formation of GLC-82 cell aggregation, and decreased the phosphorylation levels of ERK and AKT; the colony formation rate in soft agar was significantly lower in GLC-82 cells with FAK silencing than in GLC-82 cells without FAK silencing [(12.2+/-1.1)% vs. (3.6+/-0.7)%, P=0.001]. CONCLUSION: The formation of GLC-82 cell aggregation is partly mediated by FAK signal pathway, and ERK and AKT are the downstream molecules of FAK. PMID- 16219135 TI - [Biological effects of COOH-terminal amino acid deletions of hepatitis B virus X protein on hepatocellular carcinoma cell line Huh7]. AB - BACKGROUND & OBJECTIVE: The COOH-terminal amino acid deletions of hepatitis B virus X protein (HBx) commonly exist in human hepatocellular carcinoma (HCC). This study was designed to explore the biological effects of truncated HBx and wild type HBx on HCC cell line Huh7. METHODS: The mutants of HBx with 10, 20, 30, or 40 amino acids deletion at COOH-terminal (HBx3'-10, -20, -30, -40), or 37 amino acids deletion at the middle (HBx-XMR) were constructed. The recombinant truncated HBx and wild type HBx expression vectors were transfected into Huh7 cells. The integration of the exogenous vector DNA was detected by Neo gene polymerase chain reaction (PCR). The biological characteristics of positive clones were analyzed by MTT assay, colony formation assay, flow cytometry (FCM), and xenograft in nude mice. The expression of HBx3'-10, HBx3'-20, HBx3'-30, HBx3' 40, HBx-XMR and HBx was detected by immunohistochemistry. RESULTS: Huh7 cells grew faster in HBx3'-20 and HBx3'-40 groups than in HBx3'-30 group (P < 0.05). The colony formation rate was significantly higher in HBx3'-20 and HBx3'-40 groups than in HBx3'-30 and pcDNA3 groups [(17.34+/-2.77)% and (18.36+/-2.61)% vs. (7.31+/-1.44)% and (6.87+/-2.38)%, P < 0.05]. Compared with wild type HBx, HBx3'-20 and HBx3'-40 promoted more cells from G(1) phase into S phase in cell cycle [(36.96+/-1.82)% vs. (46.20+/-3.23)% and (53.99+/-4.02)% in S phase, P < 0.05], while HBx3'-10 and HBx3'-30 blocked the procedure in G(1) phase [(32.30+/ 4.32)% and (30.34+/-1.47)% in S phase]; no obvious change was found between wide type HBx group and pcDNA3 group [(38.60+/-1.15)% in S phase]. The volume of xenograft tumor in nude mice was obviously larger in HBx3'-40 group than in HBx3' 30, wild type HBx, and pcDNA3 groups [(3.19+/-0.34) cm3 vs. (1.58+/-0.27) cm(3), (1.75+/-0.15) cm3, and (1.67+/-0.12) cm3]; the tumor weight showed the same trend among the groups. CONCLUSIONS: Compare with wide type HBx, HBx3'-20 and HBx3'-40 could promote the proliferation of Huh7 cells, but HBx3'-30 has the contrary effect. HBx mutants might play a role in the development of HCC through modifying the biological functions of HBx. PMID- 16219136 TI - [CpG island methylation of E-cadherin gene promoter in gastric carcinoma]. AB - BACKGROUND & OBJECTIVE: Transcriptional silencing by CpG island methylation is now believed to be an important mechanism of carcinogenesis. E-cadherin can suppress tumor cell invasion and metastasis, and is considered as an invasion/metastasis suppressor gene. Inactivation of E-cadherin gene often occurs in poorly differentiated gastric carcinoma. This study was to investigate the CpG island methylation status of E-cadherin gene promoter in primary human gastric carcinomas and adjacent gastric tissues, and to explore the mechanism of gastric carcinogenesis. METHODS: The CpG island methylation status of E-cadherin gene promoter in 51 specimens of primary human gastric carcinoma, 37 specimens of adjacent gastric tissue, and 12 specimens of normal gastric mucosa was detected by methylation-specific polymerase chain reaction (MSP). The expression of E cadherin was evaluated by immunohistochemistry. RESULTS: The CpG island methylaion was not detected in normal gastric mucosa, but detected in 32 (62.7%) gastric carcinoma specimens and 4 (10.8%) adjacent gastric specimens. It occurred more frequently in poorly differentiated carcinomas than in well differentiated carcinomas [72.2% (26/36) vs. 33.3% (6/15), P<0.01], and it was presented at similar rates in stage T1/T2 carcinomas and stage T3/T4 carcinomas [55.6% (10/18) and 66.7% (22/33), P>0.05]. Down-regulation of E-cadherin expression was detected in 84.5% (27/32) tumor tissues with methylated CpG island and 26.3% (5/19) tumor tissues with unmethylated CpG island. No statistical correlation was found between CpG island methylaion and Helicobacter pylori infection. CONCLUSIONS: The CpG island methylaion of E-cadherin gene promoter commonly exists in gastric carcinoma, particularly in poorly differentiated adenocarcinoma. E-cadherin promoter methylation may be involved in the early process of gastric carcinogenesis, but has no correlation to Helicobacter pylori infection. PMID- 16219137 TI - [Polymorphisms of thymidylate synthase gene and correlation of its protein expression to lymph node metastasis of esophageal squamous cell carcinoma]. AB - BACKGROUND & OBJECTIVE: Thymidylate synthase (TS) is a key enzyme in DNA synthesis. The 28-bp tandem repeat in the 5'-untranslated region (UTR) of TS gene and the G/C single nucleotide polymorphism (SNP) of TS gene may modify the expression and activity of TS protein, therefore, may change the susceptibility and prognosis of tumors. This study was to explore the correlations of TS 5'-UTR polymorphism to lymph node metastasis of esophageal squamous cell carcinoma (ESCC) and the expression of TS protein. METHODS: Peripheral leucocyte DNA was extracted from 232 ESCC patients and 348 age-and gender-matched healthy controls. TS 5'-UTR tandem repeat and the G/C SNP genotype was detected by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP), respectively. TS expression in 51 specimens of ESCC was detected by SP immunohistochemistry. RESULTS: The frequencies of 3G/3G, 3G/3C, 3C/3C, 2R/3G, 2R/3C, 2R/2R, and other genotypes were 17.5%, 17.3%, 29.3%, 12.9%, 17.8%, 3.7%, and 1.5% in the healthy controls, and 16.0%, 16.0%, 29.3%, 13.8%, 17.6%, 4.3%, and 3.0% in the ESCC patients; whereas the frequencies of 3G, 3C, 2R, and other alleles were 32.8%, 47.0%, 19.5%, and 0.7% in the healthy controls, and 31.2%, 46.8%, 20.5%, and 1.5% in the ESCC patients, respectively. Compared with 3G/3G genotype, 2R/3G genotype significantly increased the risk of lymph node metastasis of ESCC [age and gender adjusted odds ratio (OR), 11.53; 95% confidence interval (CI), 2.67-49.74]. TS protein expression was significantly related to TS 5'-UTR genotype (P<0.05), but was not related to gender, age, lymph node metastasis and clinicopathologic stage. CONCLUSION: TS 5'-UTR tandem repeat and G/C SNP genotype, but not TS expression, might be a candidate molecular marker to predict lymph node metastasis of ESCC. PMID- 16219138 TI - [Expression and Significance of Notch1, P21WAF1 and involucrin in nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Abnormal expression of Notch1 protein was often found in many kinds of primary tumors, but its correlation with nasopharyngeal carcinoma (NPC) is still unclear. This study was designed to investigate the expression of Notch1 and its downstream proteins P21(WAF1) and Involucrin in NPC, and analyze their correlations with the differentiation of NPC cells. METHODS: The expression of Notch1, P21(WAF1), and Involucrin in 101 specimens of NPC and 20 specimens of chronic inflammatory nasopharyngeal mucosa were detected by SP immunohistochemistry. RESULTS: The positive rates of Notch1, P21(WAF1), and Involucrin were 100%, 90.0%, and 100% in chronic inflammatory nasopharyngeal mucosa, and were 77.2%, 89.1%, and 80.1% in NPC, respectively. The expression of Notch1, P21(WAF1), and Involucrin were significantly suppressed along with descending differentiation of NPC (P=0.000, P=0.026, and P=0.000). The positive rates of Notch1, P21(WAF1), and Involucrin were significantly higher in keratinizing squamous cell carcinoma (KSCC) than in differentiated nonkeratinizing carcinoma (DNKC) and undifferentiated carcinoma (UDC) (P<0.05), and were significantly higher in DNKC than in UDC (P<0.05). The expression of Notch1 in NPC was positively correlated with the expression of P21(WAF1) (r=0.306, P=0.002) and Involucrin (r=0.325, P=0.001). No significant correlation was found between the expression of P21(WAF1) and Involucin. CONCLUSION: The expression of Notch1, P21(WAF1), and Involucrin are closely correlated to the differentiation of NPC cells. PMID- 16219139 TI - [Establishment of cellular hypoxia model and its effects on biological behavior of gastric cancer cell line SGC-7901]. AB - BACKGROUND & OBJECTIVE: Hypoxia is an elementary characteristic of tumor microenvironment. Establishing hypoxia microenvironment in vitro will be of help in studying the effects of hypoxia on tumor cells. Previous methods of establishing hypoxic cell culture model are fussy, and researches on the effects of hypoxia on gastric cancer cells are rare. This study was to establish hypoxia microenvironment with GasPaK method in vitro, and observe the effects of hypoxia on biological features of gastric cancer cell line SGC-7901. METHODS: The hypoxia microenvironment was established by GasPaK method; the changes of PO2, PCO2, and pH in RPMI-1640 were monitored by blood gas analysis. The effect of hypoxia on cell cycle of SGC-7901 cells was analyzed by flow cytometry (FCM). The morphology of SGC-7901 cells was observed under light microscope and transmission electron microscope. Live cells were counted with trypan blue staining. The adhesiveness of SGC-7901 cells was detected by MTT assay; migration ability of SGC-7901 cells was assessed by movement experiment. RESULTS: GasPaK formed hypoxia microenvironment in 0.5-48 h with stable PO2, PCO2, and pH. When cultured in hypoxia for 16 h, no significant change in cell cycle of SGC-7901 cells was observed; morphology of some SGC-7901 cells was changed; the number of live cells didn't significantly decreased; the adhesiveness and migration ability of SGC 7901 cells were significantly enhanced. CONCLUSIONS: GasPaK method can stably establish hypoxia microenvironment with good repetition. Hypoxia has slight effect on SGC-7901 cells, and may induce morphologic change of SGC-7901 cells. PMID- 16219140 TI - [Clinical analysis of 38 cases of hepatic focal nodular hyperplasia and literature review]. AB - BACKGROUND & OBJECTIVE: Focal nodular hyperplasia (FNH) is a rare benign hepatic tumor, and its diagnosis and treatment remain controversial. This study was to summarize the clinical features of FNH, and explore its diagnosis and treatment principles. METHODS: Clinical data of 38 FNH patients, treated in our hospital from Jul. 1997 to Jul. 2004, were studied retrospectively with literature review to summarize the clinical feature, diagnosis, and treatment of FNH. All the patients were followed up till Jul. 2004. RESULTS: Among the 38 patients, 30 (78.9%) were under 40 years old, 26 (68.4%) were asymptomatic, 32 (84.2%) had normal liver function, and alpha fetoprotein (AFP) was negative in all the patients. Correct diagnosis rates were 5.3% (2/38) for B ultrasonography, 45.5% (5/11) for color Doppler ultrasound, 100% (2/2) for contrast-enhanced ultrasound, and 36.8% (14/34) for double-phase helical CT scan. Of the 38 patients, 34 (89.5%) underwent hepatic resection without any complication, 1 underwent biopsy and transcatheter arterial embolization (TAE), 2 underwent biopsy and percutaneous ethanol injection (PEI), 1 only underwent biopsy. The size of lesions didn't change after TAE or PEI. HCC appeared in 2 patients with hepatitis 69 and 32 months after the resection of FNH lesions. At the end of follow-up, 37 patients were survival, and the other one died of other disease 1 year after PEI. CONCLUSIONS: The patients with FNH usually have no clinical symptoms with good prognosis. The combination of multiple imaging measures is of help to the diagnosis of FNH. We recommend close observation for patients with confirmed diagnosis of FNH and without clinical symptoms, and recommend surgical resection for those with equivocal diagnosis, symptoms, lesion enlargement, or accompanied with hepatitis or cirrhosis. PMID- 16219141 TI - [Impact of dose boost in skull base on recurrence of stage T4 nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: The recurrence rate in skull base is high for nasopharyngeal carcinoma (NPC) patients with cavernous sinus or/and sphenoid sinus involvement. This study was conducted to explore the impact of dose boost in skull base on the recurrence in skull base and survival of NPC patients with cavernous sinus or/and sphenoid sinus involvement. METHODS: A total of 120 stage T4 NPC patients with cavernous sinus or/and sphenoid sinus involvement proved by histopathology and computed tomography (CT) were treated in our hospital from Oct. 1996 to Nov. 1998. The irradiation dose was (71.55+/-3.09) Gy in nasopharynx and (58.95+/-6.16) Gy in neck. Of the 120 patients, 27 received irradiation (6-10 Gy) in skull base after radiotherapy (boost group), 93 did not receive irradiation in skull base (control group). Fifty-three patients, 41 in control group and 12 in boost group, received cisplatin-based chemotherapy for 1-3 cycles. The survival rates and the recurrence of the 2 groups were calculated by Kaplan-Meier method and log-rank test. RESULTS: The clinical characteristics of patients were similar in the 2 groups. The 1-, 3-, 5-year skull base control rates were significantly higher in boost group than in control group (100% vs. 93.4%, 91.8% vs. 76.4%, and 86.8% vs. 65.1%, respectively, P=0.045). The 1-, 3-, 5-year disease-freely survival rates were slightly higher in boost group than in control group (85.2% vs. 75.3%, 59.3% vs. 45.2%, and 51.9% vs. 31.2%, respectively, P=0.084). The median disease-freely survival time was longer in boost group than in control group (60 months vs. 30 months). The prevalences of irradiation-induced brain damage were 7.4% in boost group and 4.3% in control group with no significant difference (P=0.514). CONCLUSION: Dose boost in skull base can reduce the recurrence of stage T4 NPC in skull base and tends to enhance the disease-freely survival rate for NPC patients with cavernous sinus or/and sphenoid sinus involvementu it is recommended to such patients. PMID- 16219142 TI - [Feasibility of thoracoscopic pulmonary metastasectomy]. AB - BACKGROUND & OBJECTIVE: With the developments in endoscopic surgery and the advances of radiological techniques, thoracoscopic pulmonary metastasectomy has been used more and more widely. This study was to evaluate the feasibility of thoracoscopic resection of pulmonary metastases. METHODS: The clinical data of 17 patients who underwent thoracoscopic pulmonary metastasectomy were retrospectively analyzed. RESULTS: All pulmonary metastases in the 17 patients were peripherally located and encountered during follow-up after treatment of primary disease. Metastases at other sites were excluded. Among the 17 cases of metastases, 10 were solitary metastasis, 4 were unilateral multiple metastases, 3 were bilateral metastases. All patients underwent thoracoscopic wedge resection. No major complications, operative mortality, and port site metastasis presented. Postoperative locoregional relapse occurred in 5 patients, in which 4 patients had multiple metastases, the rest 1 patient with soft tissue sarcoma underwent a second thoracoscopic resection of recurrent pulmonary lesion. The 1-, 3-, and 5 year survival rates were 77.3%, 53.1%, and 34.8%, respectively. CONCLUSIONS: Thoracoscopic resection is feasible and effective for patients with a solitary pulmonary metastasis, especially when the lesion is smaller than 3 cm and peripherally located. Bilateral pulmonary metastases may also be resected by primary thoracoscopic operation. PMID- 16219143 TI - [Prognostic analysis of stage III-IV non-small cell lung cancer patients treated by traditional chinese medicine]. AB - BACKGROUND & OBJECTIVE: Chemotherapy is a treatment for stage III-IV non-small cell lung cancer (NSCLC), but the efficacy is not ideal. Traditional Chinese medicine (TCM) has certain effect on NSCLC. This study was to investigate various factors that affect the prognosis of advanced NSCLC, and evaluate the role of TCM in enlonging survival time of patients with stage III-IV NSCLC. METHODS: The NSCLC patients who meet the inclusive criteria were randomized into TCM group, combination (TCM plus NP regimen) group, and chemotherapy group, and received relevant treatments. The median survival time (MST) was calculated by Kaplan Meier method. The prognosis of the patients was analyzed by COX regression method. RESULTS: A total of 294 stage III-IV NSCLC patients were enrolled, of which 99 were in TCM group, 103 in combination group, 92 in chemotherapy group. The MST were 292 days in TCM group, 355 days in combination group, and 236 days in chemotherapy group; the cumulative survival rates were 45.38%, 48.86%, and 42.17%, respectively (P>0.05). Cox regression analysis indicated that therapy, gender, disease course, erythrocyte sedimentation, KPS score, tumor size, and patient's weight were independent prognostic factors of stage III-IV NSCLC. CONCLUSION: Compare with chemotherapy alone, TCM combined with chemotherapy may prolong the survival time of stage III-IV NSCLC patients. PMID- 16219144 TI - [Bowel control of elderly low rectal cancer patients after anus-retained operation]. AB - BACKGROUND & OBJECTIVE: Though anus-retained operation has became the first choice in radical cure operation for rectal cancer, most surgeons whom fear of dissatisfied bowel control after operation recommend permanent bowel stoma in abdomen for elderly low rectal cancer patients rather than anus-retained operation. This study was to evaluate the bowel control of elderly low rectal cancer patients after anus-retained operation. METHODS: A total of 80 elderly low rectal cancer patients were divided into > or =75-year old group (39 patients) and 60-74-year old group (41 patients). Clinical data and follow-up data of the 80 patients were analyzed; bowel function and continence of the 2 groups were compared. RESULTS: The postoperative 18-month survival rate of all patients was 98.8%. The bowel control statuses of 76 patients were evaluable, of which 36 were in > or =75-year old group, 40 were in 60-74-year old group. Three months after operation, the bowel function was significantly poorer in > or =75-year old group than in 60-74-year old group (P<0.05), but the difference dissolved 6 months after operation (P>0.05). The time of recovering normal defecation frequency was slightly longer in > or =75-year old group than in 60-74-year old group (P>0.05). The prevalence of grade I incontinence 6 months after operation was significantly higher in > or =75-year old group than in 60-74-year old group (36.1% vs. 12.5%, P<0.05), but all symptoms of incontinence were relieved after treatment. CONCLUSIONS: Most elderly low rectal cancer patients could maintain bowel control after anus-retained operation. Age alone should not be a contraindication to a restorative resection for low rectal cancer. PMID- 16219145 TI - [Risk factors and prognosis of node-positive cervical carcinoma]. AB - BACKGROUND & OBJECTIVE: Pelvic lymph node metastasis is an important prognostic factor of cervical carcinoma, but few have been reported on both risk factors and prognostic factors of node-positive cervical carcinoma. This study was to evaluate risk factors, distribution pattern, and prognosis of node-positive cervical carcinoma, and explore the forecast of lymph node metastasis and appropriate treatment. METHODS: Clinical data of 205 cervical cancer patients who had undergone radical hysterectomy and bilateral pelvic lymphadenectomy were analyzed retrospectively. RESULTS: The overall prevalence of lymph node metastasis was 24.4% (50/205). Univariate analysis showed that risk factors of node metastasis were serum level of squamous cell carcinoma antigen (SCC-Ag) before treatment, clinical stage, invasive depth of cervical canal or vaginal portion of the cervix, and uterine ligaments involvement. SCC values exceeding 4 microg/L increased the risk of nodal metastasis by 4.2 folds (P<0.001, OR=4.212). Multivariate analysis showed that clinical stage and invasive depth of cervical canal were the major risk factors. The obturator and obturator fossae lymph nodes were the most frequently involved, with a rate of 48.0%. Moreover, 60.0% node positive patients had multiple sites lymph node metastases, and saltatory metastasis was found. Lymph node metastasis was closely related to deep muscularis involvement of the cervix and parametrial involvement; 72.0% nodal metastases were accompanied with deep muscularis involvement of the cervix, 90.9% uterine ligament invasions were accompanied with lymph node metastasis. The 5 year survival rate was significantly higher in the patients received postoperative radiation than in the patients didn't receive radiation (89.1% vs. 45.5%, P=0.012). CONCLUSIONS: Serum level of SCC-Ag before treatment exceeding 4 microg/L, deep muscularis involvement of vaginal portion of the cervix, uterine ligaments involvement, especially advanced stage and deep muscularis involvement of the cervical canal, are risk factors of pelvic lymph node metastasis of cervical cancer. The standard type III radical hysterectomy and bilateral pelvic lymphadenectomy should be performed to the patients with high risk of lymph node metastasis to ensure enough amplitude of parametrectomy and excision of positive nodes. When lymph node metastasis is confirmed after surgery, postoperative radiation can improve the prognosis. PMID- 16219146 TI - [Correlation of GST-pi and PCNA expression to prognosis of advanced maxillary sinus squamous cell carcinoma]. AB - BACKGROUND & OBJECTIVE: Tumor markers which relate to cell proliferation and metabolism have seldom been studied in maxillary sinus cancer. This study was conducted to identify the correlation of glutathione S-transferase pi (GST-pi) and proliferating cell nuclear antigen (PCNA) expression to prognosis of advanced maxillary sinus squamous cell carcinoma (SCC). METHODS: The expression of GST-pi and PCNA in 54 specimens of maxillary sinus SCC, 29 specimens of benign maxillary tumor, and 20 specimens of normal nasal mucosa was detected by immunohistochemistry. The correlation of GST-pi and PCNA expression to prognosis of advanced maxillary sinus SCC was analyzed by Kaplan-Meier method. The prognosis was analyzed by Cox multivariate model. RESULTS: The overexpression rates of GST-pi and PCNA were significantly higher in maxillary sinus SCC than in benign maxillary tumor and normal nasal mucosa (74.1% vs. 89.6% and 15.0%, P<0.01; 79.6% vs. 3.4% and 0, P<0.01). The 5-year survival rate was significantly higher in advanced maxillary sinus SCC patients with high expression of GST-pi than in the patients with low expression of GST-pi (34.5% vs. 21.2%, P=0.025); the difference between the patients with high and low expression of PCNA was not significant (18.0% vs. 27.0%, P=0.890). The expression of GST-pi was an independent prognostic factor of advanced maxillary sinus SCC (P=0.039, odds ratio>1). CONCLUSION: The overexpression of GST-pi is an independent prognostic factor of advanced maxillary sinus SCC, but that of PCNA isn't. PMID- 16219148 TI - [Correlation of lymphangiogenesis to progression of colorectal cancer]. AB - BACKGROUND & OBJECTIVE: Lymphatic metastasis affects the prognosis of colorectal cancer patients. The binding of vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor receptor-3 (VEGFR-3) promotes lymphangiogenesis and lymphatic metastasis. This study was to investigate the interrelation of VEGF-C, VEGFR-3 and lymphangial density (LAD), and to identify their correlations to clinicopathologic factors of colorectal cancer. METHODS: The tissue microarrays containing 105 cases of colorectal cancer and 105 cases of normal colorectal tissue were produced separately. The expression of VEGF-C and VEGFR-3 was detected by immunohistochemistry; LAD was assessed through Podoplanin immunohistochemical staining. RESULTS: The positive rates of VEGF-C and VEGFR-3, and LAD were significantly higher in colorectal cancer than in normal colorectal tissue (61.0% vs. 22.8%, P<0.01; 55.2% vs. 20.0%, P<0.01; 8.91+/-3.75 vs. 6.68+/ 1.38, P<0.01). The expression of VEGF-C had no correlation to that of VEGFR-3. LAD was significantly higher in VEGF-C-positive colorectal cancer than in VEGF-C negative colorectal cancer (10.89+/-3.36 vs. 5.83+/-1.67, P<0.01). VEGF-C was up regulated in colorectal cancer at Dukes'C stage, or with lymph node metastasis or distant metastasis; VEGFR-3 was up-regulated and LAD was higher in colorectal cancer at Dukes'C stage, or with lymph node metastasis. CONCLUSION: VEGF-C and VEGFR-3 are related with the lymphangiogenesis, lymphatic metastasis, and progression of colorectal cancer. PMID- 16219147 TI - [Expression of S100-labeled dendritic cells in glottic squamous cell carcinoma and its correlation to prognosis]. AB - BACKGROUND & OBJECTIVE: Dendritic cells (DCs) are the most important antigen presenting cells, and can synthesize and secrete S100 protein. This study was designed to investigate the expression of S100-labeled DCs in glottic squamous cell carcinoma, and analyze its correlation to the prognosis. METHODS: The expression of S100-labeled DCs in peritumoral tissues of 111 specimens of gllotic squamous cell carcinoma was detected by immunohistochemistry; 17 specimens of normal tissues were used as control. The correlation of S100-labeled DCs expression to prognosis of the 111 patients was evaluated. RESULTS: The positive rates of S100-labeled DCs were 51.35% (57/111) in peritumoral tissues of gllotic squamous cell carcinoma, and 0 in normal tissues. The 5-year survival rate was significantly higher in S100-positive group than in S100-negative group (72.31% vs. 53.91%, P=0.030). CONCLUSIONS: The glottic squamous cell carcinoma patients with obvious infiltration of S100-labeled DCs may achieve good prognosis. PMID- 16219150 TI - [Experience in treating advanced prostate cancer with bladder outlet obstruction]. AB - BACKGROUND & OBJECTIVE: The incidence and discovery rate of prostate cancer is increased in recent years; with advanced age and multiple organs dysfunction, the advanced prostate cancer patients have poor quality of life. This study was to explore suitable treatment for these patients. METHODS: A total of 80 advanced prostate cancer patients with bladder outlet obstruction were treated by transurethral electrovaporization of the prostate (TVP), plus castration and antiandrogen therapy. Preoperative individualized preparation was performed for each patient. International prostatic symptom score (IPSS), maximum flow rate of urine (Q(max)), prostatic-special antigen (PSA), and ultrasonography were measured before and 3 months after operation. RESULTS: TVP were successful in all cases. Postoperative IPSS was significantly lower than preoperative IPSS in patients with or without urine retention (13+/-3 vs. 31+/-2, 11+/-3 vs. 31+/-2, P<0.01); postoperative Q(max) was significantly higher than preoperative Q(max) in patients with or without urine retention [(19.0+/-3.3) ml/s vs. 0, (19.4+/ 2.7) ml/s vs. (8.9+/-3.4) ml/s, P<0.01]. Postoperative PSA was significantly lower than preoperative PSA [(80.4+/-133.4) mg/L vs. (0.1+/-0.4) mg/L, P<0.05]. The volume of prostate was obviously reduced. CONCLUSION: TVP plus castration and endocrine therapy is a safe and effective treatment for advanced prostate cancer patients with bladder outlet obstruction. PMID- 16219149 TI - [Diagnosis and treatment of cervical anastomotic fistula contaminating mediastinum or pleural cavity after esophagogastrostomy: a report of five cases with literature review]. AB - BACKGROUND & OBJECTIVE: A special kind of fistula will be formed when cervical gastroesophageal anastomotic fistula occurs and its contents contaminate mediastinum or pleural cavity after esophagogastrostomy. The diagnosis and treatment are difficult to made. It is very dangerous if the treatment is incorrect. This study was to analyze the causes, preventive measures, diagnosis methods and standard, and appropriate treatments of this kind of fistula. METHODS: Clinical data of 5 patients who suffered cervical anastomotic fistula contaminating mediastinum or pleural cavity after esophagogastrostomy, treated in our hospital, were retrospectively analyzed; the related literature was reviewed. RESULTS: The morbidity of this kind fistula was 1.83%(4/219) in our hospital. All the 5 patients suffered fistula 1-13 days after esophagogastrostomy; bronchopleural fistula occurred in 2 patients. The main causes of this kind of anastomotic fistula were hypertensive anastomosis, low anastomotic location, incomplete suture of thorax-esophagus outlet, defection of surgical technique, and so on. The fistula could be diagnosed correctly when the patient suffered high fever, dyspnea and thoracalgia after operation, with cervical anastomotic external fistula which could not been healed when the wound was opened, X-ray showed widened mediastinum and hydropneumothorax, and drainage, esophagoraphy and CT-confirmed anastomotic fistula. All patients were cured in a short term with sufficient drainage and irrigation, enough nutrition, appropriate use of antibiotics, and pyothorax dissection and muscle flaps transplantation for bronchopleural fistula. CONCLUSION: Most cervical anastomotic fistulas happen 1 13 days after esophagogastrostomy with critical conditions; timely diagnosis and proper treatment including operation could shorten the disease course and greatly decrease the mortality. PMID- 16219151 TI - [Selective expansion of human natural killer cells]. AB - BACKGROUND & OBJECTIVE: Natural killer (NK) cells have strong potential of killing tumor cells, but it is difficult to expand sufficient NK cells to satisfy the treatment of tumor. This study was to explore the feasibility of efficiently expand NK cells through co-culturing peripheral blood mononuclear cells (PBMCs) and Wilms tumor cell line HFWT. METHODS: PBMCs and HFWT cells were co-cultured to expand NK cells. The cytotoxicity of NK cells was measured using 51Cr release assay and crystal violet (CV) staining. The proportions of CD3(+), CD4(+), CD8(+), CD16(+), and CD56(+) cells were detected using flow cytometry. RESULTS: When PBMCs were co-cultured with irradiated HFWT cells for 10-21 days, NK cells were selectively expanded; the proportion of CD56(+)/CD16(+) cells was over 50%. The NK cells killed more than 80% of fresh HFWT cells at an effector/target ratio of 2. The expansion of NK cells required continuous stimulation of PBMCs by HFWT cells. CONCLUSION: HFWT cells can effectively stimulating PBMCs to sensitively expand human NK cells for adoptive immunotherapy of human tumors. PMID- 16219152 TI - [Correlations of centrosome abnormality and genomic instability to tumor]. AB - Genomic instability, including whole chromosome loss or gain, ploidy change, and a variety of chromosome aberration, is a characteristic of tumor cells. Centrosome abnormality is also found in several kinds of tumors. Abnormal centrosome can cause multipolar spindle formation, chromosome mis-segregation, and unequal distribution, and finally leads to cancers. Therefore, abnormal centrosome is common in tumor cells, and centrosome amplification is probably an early event in the origination and development of cancer. PMID- 16219153 TI - Signal recognition and hearing protectors with normal and impaired hearing. AB - Recognition of acoustic signals when perception is subject to interference from noise has already been extensively studied. In this study the influence of hearing protectors (HP) (plugs, muffs) and hearing loss on signal recognition is examined. Different spectrums and levels of the noise are also included. The test results are shown as the masked threshold for the signals heard and identified. In the case of normally hearing subjects a frequency-independent HP (plug) improves hearing performance, while frequency-dependent HP (muffs) tends to worsen it, especially with low-frequency noise. Hearing losses even worsen hearing performance when plugs are worn. Design suggestions are made to optimise signal recognition. Minimum signal-to-noise ratio and the use of HP are discussed. PMID- 16219154 TI - Using gas chromatography for indoor-air quality control in conservation and renovation studios. AB - Investigations were carried out in the Department of Conservation of Painting and Polychrome Sculpture and in the Screen Printing Studio of the Faculty of Art at Nicolaus Copernicus University. Concentration of the vapours of organic solvents was measured in 2 workshops: the Art Conservation Studio and the Screen Printing Studio. This study attempts to evaluate the work environment in both studios by analysing the concentration of vapour solvents over 5 to 7 years of measurements. Volatile solvents--aliphatic and aromatic hydrocarbons, alcohols, esters and ketones--were detected in investigated workplaces. These compounds have a wide range of applications in cleaning and removing old varnishes, lacquers and paints; inhalation is the main route of exposure. Vapour was collected using an active sampling method. PMID- 16219155 TI - Job level risk assessment using task level ACGIH hand activity level TLV scores: a pilot study. AB - Existing upper extremity musculoskeletal disorder analytical tools are primarily intended for single or mono-task jobs. However, many jobs contain more than 1 task and some include job rotation. This case/control study investigates methods of modifying an existing tool, the American Conference of Governmental Industrial Hygienists (ACGIH) Hand Activity Level (HAL) Threshold Limit Value (TLV), to assess the upper extremity risk of multi-task jobs. Various methods of combining the task differences and ratios into a job level assessment were explored. Two methods returned significant odds ratios, (p < .05) of 18.0 (95% CI 1.8-172) and 12.0 (95% CI 1.2-120). These results indicate that a modified ACGIH HAL TLV may provide insight into the work-related risk of multi-task jobs. Further research is needed to optimize this process. PMID- 16219156 TI - Exposure to methyl methacrylate and its subjective symptoms among dental technicians, Tehran, Iran. AB - Exposure to methyl methacrylate (MMA), total dust and health symptoms were investigated in 20 dental laboratories located in Tehran, Iran. Time-weighted average (TWA) of MMA and peak concentrations were determined, using XAD-2 tubes followed by GC-FID analysis. Total dusts were evaluated gravimetrically. Health symptoms were asked using a questionnaire. TWA for technicians with direct and indirect exposure to MMA were 327.28 +/- 79.42 and 282.9 +/- 41.84 mg/m3, respectively. Peak concentration of MMA for those technicians were 337.0 +/- 36.81 and 328.88 +/- 45.40 mg/m3, respectively. There were no significant differences between TWA of MMA and peak concentration in different weekly workdays; however, within-day variations were observed (P < .05). TWA of MMA and peak concentration correlation with the laboratory volume were 0.61-0.65. Dust exposure of technicians was 2.35 +/- 2.70 mg/m3. Cough and skin dryness were the common health symptoms. Smoking and asbestos exposure history were factors influencing cough prevalence (p < .05). It is concluded that the current Short Term Exposure Limit (STEL) is not low enough to protect technicians against the adverse effects caused by MMA. PMID- 16219157 TI - Workers' perceptions of workplace safety and job satisfaction. AB - A lot of attention has been focused on workers' perceptions of workplace safety but relatively little or no research has been done on the impact of job satisfaction on safety climate. This study investigated this relationship. It also examined the relationships between job satisfaction and workers' compliance with safety management policies and accident frequency. A positive association was found between job satisfaction and safety climate. Workers who expressed more satisfaction at their posts had positive perceptions of safety climate. Correspondingly, they were more committed to safety management policies and consequently registered a lower rate of accident involvement. The results were thus consistent with the notion that workers' positive perceptions of organisational climate influence their perceptions of safety at the workplace. The findings, which have implications in the work environment, are discussed. PMID- 16219158 TI - Evaluation of hook handles in a pulling task. AB - To evaluate the effect of handle design characteristics on subjective ratings and pulling forces, meat-hook handles with various handle shapes, sizes, and hook positions were tested in a pulling task. Finger and phalange force data measured by force sensitive resistors and subjective ratings of discomfort were also evaluated. Generally subjects preferred 37-mm double frustum, 30-mm oval handles followed by 30-mm double frustum handles, 37-mm oval, and 45-mm double frustum handles. In the analyses of total pulling force, 37- and 45-mm double frustum handles showed less required pulling force than the others. The averages of finger force contributions to the total pulling force were 27.2, 28.1, 23.9, and 20.8% in order from index to little fingers. The average of phalange force contributions were 28.8, 33.6, and 37.6% for the distal, middle, and proximal phalanges, respectively. The findings illustrate that the pulling finger forces and subjective discomfort ratings were related to the handle shape as well as handle size. PMID- 16219159 TI - An ergonomic study on posture-related discomfort among preadolescent agricultural workers of West Bengal, India. AB - In India, particularly in West Bengal, preadolescents are primarily associated with agricultural work in rural areas. Owing to poor socio-economic conditions, they are compelled to carry out a considerable number of manual, rigorous tasks in agricultural fields. The main aim of this study was to investigate postures adopted by preadolescent agricultural workers during individual agricultural activities and to analyze the causes of discomfort related to those postures. Fifty male and 50 female preadolescent agricultural workers were randomly selected and a detailed posture analysis was performed with the Ovako Working Posture Analysis System (OWAS). It was observed that those workers worked continuously in awkward postures during certain agricultural activities. Consequently they suffered from discomfort in different parts of their body. Even though they were very young, they were likely to suffer from serious musculoskeletal disorders in the future. PMID- 16219160 TI - Percutaneous exposure incidents among Australian hospital staff. AB - We investigated all reported Percutaneous exposure incidents (PEI) among staff from a large Australian hospital in the 3-year period, 2001-2003. There were a total of 373 PEI, of which 38.9% were needlestick injuries, 32.7% were cutaneous exposures and 28.4% sharps-related injuries. Nurses were the most commonly affected staff members, accounting for 63.5% of the total, followed by doctors (18.8%) and other staff (17.7%). Needlestick injuries were responsible for the majority of nurses' PEI (44.7%). Sharps injuries constituted the major category for doctors (44.3%). Most needlestick injuries (67.6%) were caused by hollow-bore needles, while the majority of cutaneous exposures involved blood or serum (55.8%). Most sharps injures were caused by unknown devices (35.9%) or suture needles (34.9%). Overall, our investigation suggests that PEI is a considerable burden for health care workers in Australia. Further research is now required to determine the relationships, if any, between workers who suffer PEI and those who do not. PMID- 16219161 TI - Ergonomics and safety of manual bag sealing. AB - A variety of seals is used to close bags. Each seal has advantages and disadvantages. For shop assistants sealing bags could be a repetitive physically demanding action. Opening and closing the bags again can cause some discomfort or annoyance for consumers. Besides, it is an activity which can endanger safety, i.e., knives being used in opening, children swallowing the systems of sealing. To prevent these problems a new sealing system was developed. In this paper the opinion of shop assistants, consumers and experts on several bag sealing systems was studied. It appeared that for sealing plastic bags, adhesive tape with paper is the best out of 4 systems, closely followed by adhesive tape. It is discussed that for the elderly, there is still room for improvement in opening bag seals. PMID- 16219162 TI - Cancer immunotherapy of targeting angiogenesis. AB - Tumor growth and metastasis are angiogenesis-dependent. Anti-angiogenic therapy may be a useful approach to cancer therapy. This review discussed tumor angiogenesis and immunotherapy of targeting tumor angiogenesis from two main aspects: (1) active vaccination to induce effective anti-angiogenesis immunity; (2) passive immunotherapy with anti-pro-angiogenic molecules relevant antibody. Evidence from the recent years suggested that anti-angiogenic therapy should be one of the most promising approaches to cancer therapy. PMID- 16219163 TI - The common gammac-cytokines and transplantation tolerance. AB - Transplant rejection, like tolerance, is a T cell-dependent event. There is compelling evidence to suggest that induction of transplant tolerance is an actively learned process in which T cells need to engage with the alloantigens in order to learn to tolerate the allograft. A family of cytokines whose receptors use the same IL-2 receptor gammac chain (also called the common gammac) plays an important role in regulating multiple aspects of the allograft response (i.e. rejection vs. tolerance). It is undeniable that gammac cytokines can drive clonal expansion and effector maturation of alloreactive T cells, and therefore, targeting such cytokines or their receptor components remains an attractive way of blocking transplant rejection. However, we just started to appreciate that gammac cytokines also regulate the acquisition of transplant tolerance via programming activated T cells for apoptotic cell death and via guiding the evolution of regulatory T cells. Thus, understanding precisely the role of gammac cytokines in regulating T cell homeostasis and T cell regulation is critically important in the induction of transplant tolerance. PMID- 16219164 TI - Achievement of cellular immunity and discordant xenogeneic tolerance in mice by porcine thymus grafts. AB - Specific cellular immune tolerance may be essential for successful xenotransplantation in humans. Thymectomized (ATX), T and NK cell-depleted immunocompetent mice grafted with xenogeneic fetal pig thymic and liver tissue (FP THY/LIV) result in efficient mouse thymopoiesis and peripheral repopulation of functional mouse CD4(+) T cell. Very importantly, the reconstituted mouse T cells are specifically tolerant to pig donor antigens. Studies demonstrated that porcine MHCs mediated positive and negative selection of mouse thymocytes in FP THY grafts, whereas mouse MHCs were involved in negative selection in grafts. Therefore, T cell tolerance to xenogeneic donor antigens could be induced by grafting donor thymus tissue. Xenogeneic thymic replacement might have a potential role in the reconstitution of cellular immunity in patients with AIDS or other immunodeficiencies caused by thymus dysfunction. PMID- 16219165 TI - Forcing tumor cells to present their own tumor antigens to the immune system: a necessary design for an efficient tumor immunotherapy. AB - The general principle for tumor cells to escape from immune surveillance is to prevent tumor antigens from being recognized by the immune system. Many methods have been developed to increase the immunogenecity of the tumor cells. The most efficient methods are able to force tumor cells to present their own tumor antigens to the immune system. Stimulating Th cells by converting tumor cells into MHC class II+/Ii- antigen presenting cells is one of the most efficient technologies. Using antisense methods, we suppress the expression of the Ii protein that normally co-expresses with MHC class II molecules and blocks the antigenic peptide binding site of MHC class II molecules during synthesis in the endoplasmic reticulum. In such tumor cells, the "unprotected" MHC class II molecules pick up endogenous tumor antigenic peptides, which have been transported into the ER for binding to MHC class I molecules. Simultaneous presentation of tumor antigens by both MHC class I and II molecules generates a robust and long-lasting anti-tumor immune response. MHC class II+/Ii- tumor cells are potent tumor cell vaccines and also cure a significant number of animals with renal and prostate tumors. We have developed analogous human gene vectors that are suitable for most patients and cancers. PMID- 16219166 TI - Activation-induced cell death in T cells and autoimmunity. AB - Activation-induced cell death (AICD), which results from the interaction between Fas and Fas ligand, is responsible for maintaining tolerance to self-antigen. A defect in AICD may lead to development of autoimmunity. During the last several years, much progress has been made in understanding the mechanism(s) of AICD and its potential role in the pathogenesis of autoimmune diseases. In this review, we summarize the most recent progress on the regulation of the susceptibility of T cells to AICD and its possible involvement in autoimmune diseases. PMID- 16219167 TI - SARS Immunity and Vaccination. AB - Severe acute respiratory syndrome (SARS) is a serious and fatal infectious disease caused by SARS coronavirus (SARS-Cov), a novel human coronavirus. SARS Cov infection stimulates cytokines (e.g., IL-10, IFN-gamma, IL-1, etc.) expression dramatically, and T lymphocytes and their subsets CD4(+) and CD8(+) T cells are decreased after onset of the disease. SARS-specific IgG antibody is generated in the second week and persists for a long time, whereas IgM is expressed transiently. The spike protein and neucleocapsid protein are most abundant in SARS-Cov and contribute dominantly to the antibody production during the course of disease. Spike protein, especially the ACE-2 binding region (318 510aa) is capable of producing neutralizing antibody to SARS-Cov. Neucleocapsid protein induces protective specific CTL to SARS-Cov. Therefore, applications with spike subunit, neucleocapsid subunit as well as inactivated SARS-Cov are three prospective vaccination strategies for SARS. PMID- 16219168 TI - Intratumoral expression of MIP-1beta induces antitumor responses in a pre established tumor model through chemoattracting T cells and NK cells. AB - Direct intratumoral introduction of therapeutic or regulatory genes is a developing technology with potential application for cancer gene therapy. Macrophage inflammatory protein-1 beta (MIP-1beta) is a chemokine which can chemoattract immune cells such as T cells. In the present study, murine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus (AdhMIP-1beta) carrying the human MIP-1beta gene. 24 h post transfection, hMIP-1beta levels reached approximately 980 pg/ml in supernatants of 10(6) hMIP-1beta-transfected CT26 cells. Moreover, the supernatants exhibited chemotactic activity for CD8(+) T cells, CD4(+) T cells, NK cells and immature DCs. Intratumoral injection of AdhMIP-1beta significantly inhibited tumor growth and prolonged the survival time of tumor-bearing mice. Intratumoral hMIP-1beta gene transfer also induced powerful tumor-specific CTL responses in vivo. The therapeutic effects of hMIP-1beta gene therapy were greatly reduced following in vivo depletion of both CD4(+) and CD8(+) T cells, but were unaffected by depletion of single T cell subsets. Immune cell depletion experiments also revealed that NK cells played an important role in hMIP-1beta-induced antitumor responses. These results suggest that intratumoral expression of hMIP-1beta has the potential effect to induce host antitumor immunity and may prove to be a useful form of cancer gene therapy. PMID- 16219169 TI - IL-6-deficient mice are susceptible to ethanol-induced hepatic steatosis: IL-6 protects against ethanol-induced oxidative stress and mitochondrial permeability transition in the liver. AB - Interleukin-6 (IL-6)-deficient mice are prone to ethanol-induced apoptosis and steatosis in the liver; however, the underlying mechanism is not fully understood. Mitochondrial dysfunction caused by oxidative stress is an early event that plays an important role in the pathogenesis of alcoholic liver disease. Therefore, we hypothesize that the protective role of IL-6 in ethanol induced liver injury is mediated via suppression of ethanol-induced oxidative stress and mitochondrial dysfunction. To test this hypothesis, we examined the effects of IL-6 on ethanol-induced oxidative stress, mitochondrial injury, and energy depletion in the livers of IL-6 (-/-) mice and hepatocytes from ethanol fed rats. Ethanol consumption leads to stronger induction of malondialdehyde (MDA) in IL-6 (-/-) mice compared to wild-type control mice, which can be corrected by administration of IL-6. In vitro, IL-6 treatment prevents ethanol mediated induction of reactive oxygen species (ROS), MDA, mitochondrial permeability transition (MPT), and ethanol-mediated depletion of adenosine triphosphate (ATP) in hepatocytes from ethanol-fed rats. Administration of IL-6 in vivo also reverses ethanol-induced MDA and ATP depletion in hepatocytes. Finally, IL-6 treatment induces metallothionein protein expression, but not superoxide dismutase and glutathione peroxidase in cultured hepatocytes. In conclusion, IL-6 protects against ethanol-induced oxidative stress and mitochondrial dysfunction in hepatocytes via induction of metallothionein protein expression, which may account for the protective role of IL-6 in alcoholic liver disease. PMID- 16219170 TI - Novel SLA-DR alleles of three Chinese pig strains and the related function in human T cell response. AB - To elucidate the structures of SLA-DR (swine leukocyte antigen DR) genes of three Chinese pig strains (Gz, Bm and Yn), the SLA-DRA and SLA-DRB cDNA were amplified by RT-PCR and subjected to determine the sequences. The whole structures of SLA DRA alleles are identical among three strains, consisting of 759 nucleotides including an open reading frame (ORF), and are shared with those reported from NIH minipigs SLA-DRA(c) and SLA-DRA(d). The same length of the ORF-containing SLA DRB genes of three Chinese pig strains was also identified. They are composed of 801 nucleotides encoding a xenogeneic antigen molecule of 266 amino acid residues. The nucleotide sequences of the SLA-DRB genes, however, are different when compared either among the three strains or with the published data of SLA DRB sequences, which allowed our novel SLA-DRB alleles receiving their accession numbers AY102479, AY102480 and AY102481 from the GenBank. This study further reveals that the phylogenic homologies of MHC DR or DR-like genes in structures of nucleotides and deduced amino acids between Chinese pigs (SLA) and human (HLA DRB1*0901) are better than those between pigs and mice (H-2(b)Ebeta). High similarities were also found for DRalpha-DRbeta heterodimers between Chinese pigs and human in terms of amino acids sequences critical for binding with human CD4 coreceptor molecule, which are better than those between SLA-DR and H-2 I-E molecules. A functional test indicated that, by cotransfection with Bm-DRA and Bm DRB genes, the Bm-DR molecule-expressed L929 cells could stimulate human T cells quite well in a xenogeneic reaction in presence of human APCs. PMID- 16219171 TI - The levels of plasma IL-1beta, IL-6 of C57BL/6J mice treated with MPTP and brain lateralization. AB - The work is to explore the relationship between the levels of cytokines (IL-1beta and IL-6) in C57BL/6J mice treated with MPTP and brain lateralization. By using paw preference test, right-pawed, left-pawed mice models were established. Following single injection of 1-methyl-4-pheny1-2, 3, 6-tetrahydropyrid (MPTP) (40 mg/kg) to impair dopaminergic neuron, enzyme linked immunosorbent assay (ELISA) kits were used for detection of plasma levels of cytokines. The results showed that in saline treated C57BL/6J mice (control), there was no obvious difference observed between left-pawed and right-pawed mice in plasma levels of IL-1beta and IL-6. In MPTP treated mice, there was no difference between level of IL-1beta in left-pawed mice and that in right-pawed ones in statistics, that is, they were increased on day 1 and day 3, but decreased on day 6. The plasma level of IL-6 was lower in left-pawed than that in right-pawed mice (p < 0.005) after MPTP treatment. On day 1 and day 3, the level of IL-6 was almost the same as control; on day 6, it was significantly increased, higher than that of control (p < 0.001) in left-pawed mice. While in right-pawed mice, on day 1 and day 3, it was no different from control, too. And on day 6, it significantly increased in compared with control (p < 0.005). In conclusion, the level of plasma IL-6 of C57BL/6J mice treated with MPTP increased. The variation of IL-6 was correlated to brain lateralization. PMID- 16219172 TI - Analysis of the expression of Fas, FasL and Bcl-2 in the pathogenesis of autoimmune thyroid disorders. AB - To investigate the expression of apoptosis-related protein (Fas, FasL, and Bcl-2) in the pathogenesis of autoimmune thyroid disorders (ATDs), immunohistochemical staining was performed on 20 Hashimoto's thyroiditis (HT), 20 Graves' disease (GD), and 20 thyroid follicular adenoma (TFA, as control). All the cases expressed Fas, mainly on the cell surface and cytoplasm. FasL was found in 17 cases of the TFA. Bcl-2 was detected in 15 cases of HT, 19 of GD and 17 of TFA. In TFA, a moderate Fas expression and a minimal or no FasL expression was detected on follicular cells. In HT, the follicles adjacent to infiltrating lymphocytes showed increased levels of Fas and FasL expression. A weaker staining of Fas and FasL was exhibited on infiltrating lymphocytes than on thyrocytes. In a comparison of GD with HT, thyrocytes and lymphocytes showed similar Fas staining, but for FasL the staining was rather weaker in HT. The expression of Bcl-2 was nearly identical in GD and TFA, but much weaker on the follicular cells in vicinity of lymphocytes and on the lymphocytes located in germinal centers of HT tissues. The expression of Fas, FasL, Bcl-2 in Hashimoto's thyroiditis and Graves' disease were almost same. FasL strong expression and Bcl-2 weak expression on the follicles in HT may induce apoptosis. These results provided evidence for expression of Fas, FasL and Bcl-2 in the pathogenesis of autoimmune thyroid disease. The lymphocytes seem not to be directly engaged in the process via their own FasL, but they may provide some cytokines that, in turn, upregulate Fas and/or FasL expression to induce apoptosis. PMID- 16219173 TI - Combined transfection with EBV-specific epitopes and HLA-A2 genes is more effective than separate transfection in promoting CTL lysis against nasopharyngeal carcinoma. AB - To augment specific cytotoxic T lymphocyte (CTL) lysis is a promising strategy for cancer therapy. In this study, we examined the boosting effect of CTLs upon autologous lymphoblastoid B cell lines (LCLs) transfected with diverse plasmids, to explore the possible CTL-based immunotherapy of nasopharyngeal carcinoma (NPC). FCM analysis displayed rather high ratio (>30%) of successfully transfected LCLs by utilizing the DMRIE-C kit. CTL assays demonstrated that substantially higher ratio of CTL specific lysis was observed upon the LCLs transfected with both expression vectors encoding EBV-specific epitopes and their presentation molecule HLA-A2, in contrast with those transfected separately. By transfecting the vector encoding HLA-A2 alone, only the LCLs of HLA-A2(+) donors elicited markedly higher CTL lysis. CTL assays also showed that there existed no marked differences upon transfection by either different vectors (pcDNA3, pNGVL3 or pNGVL3-hFlex), or different EBV-derived peptides (LMP(2)Pep1 or LMP(2)Pep2), or with or without the doubled DNA sequence encoding peptides. This study indicated a promising immunotherapy strategy on NPC through boosting and eliciting the EBV-specific CTL activation by transferring vectors encoding both EBV-specific epitopes and their presentation molecule HLA-A2 into autologous LCL, the presentation cells of MHC/peptide tetrameric complex. PMID- 16219174 TI - [Hospital nurse staffing and quality of care]. PMID- 16219175 TI - [The world of nursing burnout. A literature review]. AB - The aim of this paper is to photograph, through a literature review, the burnout and all its related aspects in the world of nursing. We have selected 40 papers with a specific valence which frame and study the nursing burnout in an appropriate way. This topic has been studied in all of the continents, apart from South America. The burnout has been subject of analysis in all the departments, whom oncology and psychiatry result the ones with the highest frequency of studies. In this paper all the studies have been divided into three categories according to their objective: studies on aspects related to the burnout (57,5%), comparative studies (25%), studies on the burnout level (17,5%). From the studies concerning the related aspects comes out that the burnout is strongly related to the coping strategies, the stress index and the personal accomplishment and these factors are highly predictive. Beside that, the data show some disagreements concerning the relation between burnout and sex and burnout and age. The comparative studies show that nurses are, between health care staff, the ones which run the highest risk of burnout, especially the oncologic nurses and nurses working with AIDS patients. This literature review, summing up which steps the research has done and which are still to do, reflects the strong importance of the problem called burnout and the aspects related to it inside the world of nursing. This review underlines the importance of an insertion of support group inside the clinical practice to avoid the risk to have sick persons who take care of other patients. PMID- 16219176 TI - [Continuity of care: community care in Italy and Sweden. A comparison]. AB - One of the objectives of the Health 21 document, approved by the World Health Association, European Region, is to create by the year 2010 an integrate sanitary system, where the population of the region count on a better access to the health services aimed at the family and the community. In accordance with the tendencies of the European nation, which are more and more open to confrontation, the research has, as main objective, the parallel analysis of the welfare continuity and integration of the Italian Nursing Home Care Service compared to the Swedish Health Care Service. The research has been structured from a sanitary accreditation project that the Veneto Region has started with the collaboration of the Ottawa Council On Health Services Accreditation (Cchsa). The analysis puts in evidenced the completion of the offer of the territory home services present in Padua. The comparison with the Swedish situation however puts in evidence some gaps and some possible developments of the Italian service involving above all the complete transcription of the patient information and the continuity of its communication, in particular in the nursing field. Another possible development of the Italian service concerns the method continuity, between the working units themselves and with the territory, which could be achieved using common nursing guide lines and protocols to manage standard caring situations. From the analysis of the european services it results that it is the nurse who has the most relevant role for the care quality improvement. PMID- 16219177 TI - [Bioethics and care: from human rights to quality of care]. AB - The pluralist nature of bioethics and the fact that it spans various branches of science places it in a privileged position to tackle ethical problems that accompany human choices in situations that influence the conditions of living beings. Clearly, issues such as health and care form part of the bioethical sphere of enquiry and it is obviously of fundamental importance that these subjects receive a suitable degree of attention within a branch of science that has always drawn on the contributions of other human sciences. Bioethics looks at the great questions posed by moral principles and seeks to apply them to present concerns. PMID- 16219178 TI - [Some considerations on nursing students clinical practice]. AB - In a nurse future practical work, which is rapidly developing and changing, ethics will play an important role. A nurse student needs her/his own experience from patient care as well as training in reflecting upon ethical matters to manage future work situations. The aim of study behind this article was to find the ethical problems that engage students, how they can correlate theoretical knowledge with practical experience and what insights in ethical they may achieve by reflecting on the experiences. PMID- 16219182 TI - [Paracetamol to children]. PMID- 16219179 TI - [Tables of Moses: nursing research project]. AB - A review of literature and in others realities of our country has been done to verify the existence of welfare procedure in stomatherapy. It has been pointed out that the behaviours are not supported by evidence and that the given performances are not measurable. It borns the idea to start a nursing research finalized to define care standards procedures supported by evidence. MATERIALS AND METHOD: Once the project group is formed with nurses, ET nurses and surgeons, the research is structured in the following points: predisposition of nursing protocols; predisposition of evaluation tools for each protocol; experimentation of the tools with 150 national investigators specifically trained; gathering, elaboration and interpretation of the data; on 13.172 forms picked up, 2.034 has been regularly fulfilled; distribution of the knowledge through a text, the associative magazine, presentation at courses and associative congresses; Research development: the critic results analysis has permitted to recognize areas on which continue the research. CONCLUSION: Data confirms the validity of specifics check-lists and protocols even if the research says the achievement of care standards results through codified performances, to disadvantage of a case control way for example. PMID- 16219183 TI - [Paracetamol dosage for children--different from that for adults]. AB - Paracetamol (acetaminophen) has become the most widely used antipyretic and analgesic drug for children in Denmark. However, there is a wide discrepancy between the consumption of the drug and our knowledge of children's pharmacokinetics and pharmacodynamics. Dosage, therapeutic intervals and toxic potential are different for children and adults. In this review we look at the published guidelines for treating children with paracetamol and the evidence supporting them. PMID- 16219184 TI - [The effect of acupuncture on musculoskeletal disorders]. AB - It is estimated that between 1,500 and 2,000 Danish doctors are using acupuncture in their daily practice, in particular for diseases in the locomotor system. The purpose of this paper is to clarify what acupuncture's possible mode of action is and for which diseases within the locomotor system acceptable evidence exists for its use. The author concludes that reasonable scientific evidence exists for the alleviation of myogenic facial pain, neck pain, shoulder pain, elbow pain, low back pain and knee pain. Moreover, the author points out that acupuncture is a very safe technique in the hands of a qualified doctor with special training in the discipline. PMID- 16219185 TI - [Supervision--why?]. AB - In their work, doctors are exposed to strong emotions on the part of their patients. They need to learn to listen and not interrupt them; in general, after each consultation they focus on whatever was not accomplished. Therefore doctors need a secure environment in which they can examine common work conditions, as well as their own strengths and weaknesses, with colleagures in order to discover new ways of doing things. Based on this observation, it is a very positive development that supervision has become a part of the new specialist training for general practice. PMID- 16219186 TI - [Communication skills training for medical students: from the simple to the complex]. AB - Communication skills training at the Medical School of Copenhagen University aims to teach students basic skills in both interviewing and giving information. The article describes a method whereby the students follow a general communication skills model for giving information. The training is conducted with simulated patients (actors). It begins with providing information about simple diagnostic or operative procedures and routine tests. These skills are also practiced in clinical courses on real patients, under supervision. Later on, the students are trained in breaking bad news, but with simulated patients only. PMID- 16219187 TI - [Suicide attempters' contacts with the primary health care system]. AB - INTRODUCTION: We examined suicide attempters' use of health insurance services as part of an effort to prevent suicidal behaviour. MATERIALS AND METHODS: The research project was a register-based case control study, where cases were persons residing in the County of Funen, Denmark, who made a suicide attempt resulting in contact with a hospital. Controls were matched with respect to gender, age and county of residence on the date of the first registered suicide attempt (index attempt). Data about services paid for by the public health insurance program were collected from the Prevention Register. The data were analyzed using conditional logistic regression. RESULTS: In the calendar year prior to an index attempt, suicide attempters consulted health professionals, i.e., general practitioners, psychologists or psychiatrists, more often. Gender and age differences were small. DISCUSSION: Suicide attempters consult health insurance professionals, especially GPs, more often, and GPs' specific role in the prevention of attempted suicide is discussed. PMID- 16219188 TI - [Five years (1995-2000) of coronary artery bypass surgery at the Odense University Hospital]. AB - INTRODUCTION: A large-scale Danish study on late outcomes after coronary artery bypass surgery (CABS) has never been done. We therefore did a retrospective study and follow-up of 2,333 patients, accounting for 2,361 consecutive CABS operations over a five-year period. MATERIALS AND METHODS: The material consisted of 23.5% women and 76.5% men. The patients' median age was 63 years (32-85 years). The rate of response to the questionnaire was 95.3%. The median observation time was 26 months (3-63 months). In 94% of the patients, primary CABS had been undertaken, and 6% were reoperations. Ninty-three percent were elective operations, and 7% were acute. The risk score estimated 20% of the patients to be in high-risk groups, 22% in the 5% group and 53% in the low-risk group. There was three-vessel disease in 68% and left main stenosis in 24%. The EF was normal in 73% of the patients, while 4% had EF <30%. RESULTS: The total early mortality rate was 3%, the primary elective CABS rate was 2.6%, and we found a statistically significant higher mortality for elective reoperations (5.9%) and for acute CABS (7%). Comparison with the Danish background population indicates a better survival rate for the CABS patients. One-third developed atrial flutter or atrial fibrillation. Reoperation for bleeding was done in 6.9% and for mediastinitis 1.9%. About 10% were readmitted because of various complications. In the observation period, 99 patients (7,2%) died. Patients with EF <30% and those undergoing reoperations had a statistically significant higher late mortality rate. About 20% suffered from recurrent angina in the observation period, one-fourth had wound healing of the graft leg and 96% did not regret the operation. DISCUSSION: Apart from the rather high reoperation rate because of bleeding and the prolonged healing time of the graft leg, these results as well as the prognostic value of CABS in selected patient groups justifies the coronary artery bypass graft operation as a good treatment for angina pectoris. PMID- 16219189 TI - [Double-blind randomized trial of the effect of Bidro on hay fever in children]. AB - INTRODUCTION: The natural product royal jelly (trade name Bidro) is widely used for the treatment and prevention of allergic symptoms. Several case reports suggest marked clinical benefits. Our aim was to assess whether treatment with royal jelly modifies the development of allergic hay fever symptoms in children with pollen allergy. MATERIALS AND METHODS: Eighty children aged 5-16 years with hay fever symptoms due to birch, grass and/or mugwort participated in a placebo controlled, double-blind, randomized, controlled trial. Treatment with royal jelly or placebo was administered three to six months before and throughout the pollen season. RESULTS: The primary outcome was the occurrence of symptoms of rhinitis and conjunctivitis in the pollen season. A secondary outcome was symptom severity, as measured by symptom score, by visual analog scale (VAS) and by the need for additional hay fever treatment. Sixty-four children completed the study; 34 of them had been treated with royal jelly and 30 with placebo. All of the patients in both groups developed hay fever symptoms during in the pollen season. The severity of hay fever symptoms was similar in the two groups (VAS = 1.6 in the placebo group and 1.4 in the royal jelly group; the mean number of combined daily nasal symptoms was 2.83 in the placebo group and 2.82 in the royal jelly group). The need for additional hay fever treatment was similar in the two groups. DISCUSSION: We conclude that royal jelly has no effect on the occurrence of hay fever in the pollen season, nor does it modify the severity of hay fever symptoms. PMID- 16219190 TI - [Is guideline-defined asthma control achievable?--secondary publication. The Gaining Optimal Asthma ControL (GOAL) Study]. AB - In this study, the percentage of patients with uncontrolled asthma who can achieve guideline-defined asthma control was assessed in 3,421 patients. Significantly more patients (71%) treated with salmeterol-fluticasone achieved asthma control than did patients receiving fluticasone alone (59%). The patients treated with salmeterol-fluticasone achieved asthma control more rapidly and at a lower dose of inhaled corticosteroid than did patients treated with fluticasone alone. Achievement of asthma control was associated with marked improvements in exacerbation rates and quality of life. Thus guideline-defined asthma control can be achieved in the majority of patients and should be the goal of asthma treatment. PMID- 16219191 TI - [Central fat deposits contribute to atherosclerosis and insulin resistance and peripheral fat deposits protect against atherosclerosis and insulin resistance in postmenopausal women--secondary publication]. AB - An analysis of 290 postmenopausal women, subdivided into four distinct body fat distribution groups, showed that peripheral fat mass (PFM) confers insulin sensitizing and anti-atherogenic effects. PFM contributes to circulating adiponectin, an adipocyte-derived hormone with insulin-sensitizing, anti inflammatory, and anti-atherogenic effects. In generally obese women, the secretion of adiponectin by PFM provides effective anti-atherogenic protection. In postmenopausal women, body fat distribution has more critical implications for metabolic and cardiovascular risk than overall obesity per se. PMID- 16219192 TI - [Small-intestine tumor with ectopic pancreas tissue and acute pancreatitis]. AB - Ectopic pancreas tissue is usually a rare and incidental finding during surgery. Reports of discovery during surgery vary between 0.17% and 0.83%. We report on an 86-year-old woman admitted to the surgical ward with abdominal pain. Due to suspicion of a perforated gastric ulcer, an exploratory laparatomy was performed. An 8 x 5 cm tumor was found in the ileum 10 cm from the ligament of Treitz. Microscopic examination revealed, surprisingly, pancreatic tissue in the submucosa of the ileum with inflammation and transmural perforation. There was no indication of malignancy. PMID- 16219193 TI - [Picture of the month: bladder calculi]. PMID- 16219194 TI - [Colon ileus caused by endometriosis]. AB - A case history of acute obstruction of the sigmoid colon caused by endometriosis in a 35-year-old woman is described. Endometriosis occasionally causes relative stenoses of the bowel, whereas total obstruction is rarely seen. Obstruction of the sigmoid colon is often caused by malignancy, but sometimes other causes must be considered. PMID- 16219200 TI - [Generic preparations]. PMID- 16219201 TI - [How will the treatment of patients with liver diseases become evidence-based?]. PMID- 16219210 TI - [Palliative treatment of malignant obstruction of distal part of the ventricle or duodenum: large incision, small incision or no incision at all?]. PMID- 16219212 TI - [Propofol infusion syndrome in children]. AB - Propofol infusion syndrome, characterised by cardiovascular collapse, metabolic acidosis, hepatomegaly, renal failure and rhabdomyolysis, is a rare and often fatal syndrome seen in critically ill children undergoing long-term propofol sedation. Propofol may impair mitochondrial free fatty acid metabolism, resulting in an imbalance between energy demand and utilisation and thus compromising cardiac and peripheral muscle cell function. Propofol should be used with caution for sedation in critically ill children, as well as for long-term anaesthesia in otherwise healthy children, and doses exceeding 4 mg/kg/h should be avoided. PMID- 16219211 TI - [Magnetic resonance imaging (MRI)-directed focussed ultrasound. Methods and applications in oncological treatment]. AB - Focussed ultrasound is the only known technology that allows non-invasive local hyperthermia. Since MRI may be employed to obtain real-time temperature maps non invasively, the combination of these two technologies offers great advantages, specifically aimed towards oncological studies. Real-time identification of the target region and accurate control of the temperature evolution during the treatment are now possible. Thermal ablation of pathological tissue, local drug delivery using thermosensitive microcarriers and controlled transgene expression using thermosensitive promoters have recently been demonstrated with this unique technology, and based on these experiments the combination of focussed ultrasound and MRI thermometry holds promise for future oncological diagnostics and treatment. In this paper we review some of the recent methodological developments as well as experimental and first clinical studies using this approach. PMID- 16219213 TI - [The role of hepatobiliary scintigraphy in neonates with persistant jaundice]. AB - Hepatobiliary scintigraphy of infants being examined for persistant neonatal jaundice is best carried out using mebrofenin, a 99m Tc-labeled IDA derivative. This article summarizes the most frequently occurring etiologies of persisting neonatal jaundice and the diagnostic work-up of this condition. Perinatal obliteration of extrahepatic bile ducts requires immediate surgery, whereas other causes of persisting jaundice may be treated medically and/or conservatively. PMID- 16219214 TI - [Self-expanding metal stents as palliative treatment of a malign obstruction in the distal part of the ventricle or duodenum]. AB - INTRODUCTION: Self-expanding metal stents (SEMS) have emerged as a simple therapeutic option for the palliation of patients with non-resectable malignant gastric outlet obstruction. We present our results from a three-year period. MATERIALS AND METHODS: Twenty-nine patients with obstruction from tumors in the pancreas (15), bile ducts (3), stomach (9) or transverse colon (2) underwent palliative stenting with a 9-cm-long, 22 mm Wallstent under general anaesthesia. Insertion of the SEMS was done under endoscopic and fluoroscopic control. Biliary stents were implanted prior to or simultaneously with the duodenal stent in eight patients. Seven were covered 6-cm-long, 10 mm Wallstents. Two patients had biliary stents implanted 12 and 262 days, respectively, after the duodenal stent by "rendezvous" technique. RESULTS: The stent deployment was successful in all patients. There were no procedure-related complications, but one patient died of cardiac arrest 12 hours after the operation. Obstruction was relieved in all patients, and an exclusively oral diet was possible for 23 of them. Seven patients with rapid progression of the disease stayed in hospital and died 0-16 days after the procedure. The median length of stay in hospital after the procedure was 2 days (1-32 days), after which the patients stayed at home for 40 days (2-270 days). The overall median survival time was 47 days (median, 0-274 days). There were no late complications (stent migration or perforation), but two patients needed an overlapping stent due to tumor overgrowth. DISCUSSION: Duodenal stents effectively resolve the obstructive symptoms of gastric outlet obstruction. There are few procedure-related complications, and the vast majority of patients can leave hospital and spend the short time left to them at home. PMID- 16219215 TI - [Implementation of laparoscopic resection of the sigmoid colon, initial results and experiences]. AB - INTRODUCTION: In recent years, the enthusiasm for laparoscopic surgery has been immense. In Denmark, laparoscopic surgery of the colon has been performed in only a few cases each year. In other countries, laparoscopic surgery foreign diseases of the colon is well established. Laparoscopic surgery for malignant diseases of the colon has been awaiting the results of studies of the oncological results. Now several randomised studies have demonstrated oncological results equal to or better than those of open surgery. The aim of this study was to show the initial experience of laparoscopic surgery of the colon in a Danish hospital surgical department. MATERIALS AND METHODS: Forty patients were included in the study. They had a planned laparoscopic resection of the sigmoid colon in the period from January 2004 to February 2005. RESULTS: Thirty-six patients had the operation done laparoscopically. Four operations were converted to open operations. Four patients had major complications. The median postoperative time in hospital was three days and five days for malignant and benign disease, respectively. In the case of malignant disease, the median number of lymph nodes in the specimen was eight, and the median distance from the resection line to the tumour was 4.5 cm. CONCLUSION: Our results seems to show that if with sufficient laparoscopic expertise, laparoscopic resection of the colon for both benign and malignant diseases can be performed with the advantage of earlier convalescence and fewer days in hospital. The histopathology findings in the case of malignant disease do not differ from those of open surgery. PMID- 16219216 TI - [Iodine uptake and the effect of radioiodine treatment--status after the addition of iodine to salt]. AB - INTRODUCTION: Iodine uptake and the effect of radioiodine treatment of nontoxic goiter were investigated after the introduction of iodine to salt. MATERIALS AND METHODS: A retrospective study was done of 61 patients (48 women and 13 men with a median age of 56 (range 28-78)) with nontoxic goiter, examined by 24-hour radioiodine uptake. 42 of the patients were treated with radioiodine (median 600 MBq, range 200-600 MBq). The results were compared with those from a study done 10 years ago in the same area. RESULTS: Iodine uptake decreased significantly, from 40% (range 16-67%) in 1981-1993 to 25% (range 13-62%) in 2001-2003. 38 of the 42 patients were examined, and of these 34 (90%) had benefited from the treatment (53% of the 38 patients showed a complete effect) as estimated by the patient and the physician, based on the patient's original symptoms and the physician's clinical judgement. Four patients developed hypothyroidism, one radiation-induced thyroiditis and three Graves" disease. DISCUSSION: Iodine uptake is significantly lowered after the addition of iodine to salt. Radioiodine therapy of nontoxic goiter will continue to be effective. PMID- 16219217 TI - [Efficacy and safety of laser-assisted in situ keratomileusis for myopia]. AB - INTRODUCTION: The aim of this study was to assess the efficacy, frequency of re operation and type and frequency of complications after laser-assisted in situ keratomileusis (LASIK) treatment for moderate to high levels of myopia. MATERIALS AND METHODS: 357 eyes from 190 patients were reviewed. The mean of the preoperative correction was -8.38 D (-1.75; -16 D) sph. (spherical), -1.00 D (0; 4.5 D) astigmatism. 15% (55 eyes) were re-treated. The mean of the correction before re-treatment was -2.78 D (+2.25; -11.00 D) sph., -0.43 D (0; -2.00 D) astigmatism. RESULTS: The three-month follow-up rate was 96% for primarily treated eyes and 99% for re-treated eyes. Where full correction was intended, 53% of the primarily treated and 93% of the re-treated eyes achieved an uncorrected visual acuity of > or =0.5. The refractive error for myopia less than -8 D was 0.80 D +/- 0.83 D and -1.37 D +/- 1.33 D for myopia -8 D and above after one treatment and -0.47 +/- 0.66 D after re-treatment. 1.2% (4) of the primarily treated and 3.9% (2) of the retreated eyes lost more than two lines in best spectacle-corrected visual acuity (BCVA). CONCLUSION: LASIK treatment for moderate to high levels of myopia leads in all cases to a reduced degree of myopia. There is a significant degree of unreliability on the accuracy of the treatment with a tendency to under correction. Postoperatively, many patients still have to use correction for distance vision. Significant loss of visual acuity occurs infrequently. PMID- 16219218 TI - [Street prostitution and drug addiction]. AB - INTRODUCTION: Street-based prostitution accounts for 10% of the prostitution activity in Denmark, mainly involving female drug addicts. We studied a group of women with a common history of substance abuse and their comparative psychosocial characteristics, correlated with whether they had previously been a prostitute or not. Their psychic symptoms were evaluated and compared with those of controls. MATERIALS AND METHODS: 27 females receiving maintenance treatment for substance abuse completed a questionnaire dealing with their social background, substance abuse profile, and history of sexual abuse and prostitution, as well as their current health status, including SCL-90. The scores were compared to those of a control group of an age- and gender-matched Danish standard population. RESULTS: Neglect in childhood and adulthood corresponded to international findings. 14 of the women had previous sex-trading experience, and early use of heroin and cocaine was a predictor for starting a career in prostitution. The SCL-90 scores for the dimensions of somatization and depression were significantly higher for drug-abusing women in general than in the control group. The scores of drug abusing former prostitutes were similarly significantly higher on most of the dimensions except the hostility dimension when compared to those of drug-abusing women who had never been involved in prostitution. Rape and domestic violence were characteristic phenomena among drug-abusing prostitutes (p < or = 0.05). DISCUSSION: Victimization during childhood and adulthood constitutes a serious risk for generating social vulnerability through drug addiction and prostitution. Various psychosocial stress factors among street-based prostitutes indicate the need for broader psychiatric approaches in Danish drug addiction maintenance programmes. PMID- 16219219 TI - [Imiquimod treatment of lentigo maligna to dermoscopic and histologic clearance]. AB - Imiquimod is an immune system-modifying drug that switches the immune system to a Th1 response, enabling it to defeat virus-infected and neoplastic cells. We report on a 94-year-old woman with a large lentigo maligna (LM) on her cheek whom we treated with imiquimod 5% cream. Dermoscopy was used to identify the most suspect areas of the lesion for biopsy and in follow-up. By week 19 the lesion was cleared, and no remnants of the lesion were seen by dermoscopy or histology. Published LM cases treated with imiquimod are reviewed. PMID- 16219220 TI - [Inflammatory pseudotumour of the liver]. AB - A case study of inflammatory pseudotumour of the liver (IPL) is related. The patient presented with symptoms, biochemistry and ultrasonic findings highly indicative of a malignant disease. However, a core needle biopsy revealed inflammatory pseudotumour of the liver, an infrequent diagnosis with only about 200 cases to be found in the literature. This case illustrates the importance of core biopsy from focal processes, since IPL is a benign condition with a strong tendency to spontaneous regression. PMID- 16219221 TI - [Dilated cardiomyopathy as a side effect of treatment with infliximab]. AB - We report a case of heart failure after infusion of Remicade (infliximab) in a patient without heart insufficiency. The patient, a 25-year-old woman with inflammatory bowel disease, developed new-onset heart failure after receiving TNF antagonist therapy. After the treatment was discontinued and heart failure therapy was started, the patient improved significantly. The U.S. Food and Drug Administration has published 47 case reports of heart failure after therapy with a tumor necrosis factor antagonist, where 38 patients developed new-onset heart failure and 9 patients experienced heart failure exacerbation. There are no published reports of heart failure after TNF antagonist therapy in the Danish literature. PMID- 16219232 TI - [C vitamin deficiency in alcoholics]. PMID- 16219241 TI - A retrospective 5-year follow-up study of two different titanium implant surfaces used after interpositional bone grafting for reconstruction of the atrophic edentulous maxilla. AB - BACKGROUND: Long-term comparative follow-up studies of dental implants placed in augmented bone are rare. Variations in design and surface roughness have been found to be important for bone integration of implants. However, there is no clinical evidence that such variations lead to an improved clinical outcome. PURPOSE: To compare two different implant systems used after interpositional bone grafting of the severely resorbed maxilla with a modified augmentation technique using fibrin glue. MATERIALS AND METHODS: Twenty-two consecutive patients presenting with severe maxillary atrophy underwent reconstruction with Le Fort I osteotomies and interpositional bone grafting. Before placement of bone blocks, the floors of the maxillary sinuses were packed with bone chips mixed with a fibrin glue, to stabilize the graft. After 6 months of graft healing, the first 11 consecutive patients received Branemark System implants with a turned surface (Nobel Biocare AB, Goteborg, Sweden). The following 11 consecutive patients were treated with Astra Tech implants with a blasted titanium surface (Astra Tech AB, Molndal, Sweden). All patients received fixed prostheses. Marginal bone resorption and donor and recipient site morbidity were evaluated. All patients were clinically and radiographically observed throughout 5 years of functional loading. RESULTS: In the Branemark group, 11 (13%) of 84 placed implants were lost, compared to 4 (5.5%) of 72 placed implants in the Astra Tech group. The difference was not significant. All patients retained fixed constructions after 5 years of loading. The mean marginal bone loss was 2.3+/-0.8 mm (range, 0-5.0 mm) in the Branemark group and 2.4+/-1.4 mm (range, 0-7.0 mm) in the Astra Tech group although again no statistical difference was found. A larger number of implants in the Astra Tech group had a marginal bone resorption of>or=3 mm, and implant success in that group was lower than in the Branemark group (52% vs 70%). CONCLUSION: In this study, reconstruction of the severely resorbed maxilla with Le Fort I osteotomy, interpositional bone grafting, and delayed placement of dental implants was found to be a predictable long-term procedure. Although more implants with a turned surface were lost during the follow-up period, there were no statistically significant differences between turned and titanium blasted implants. PMID- 16219242 TI - Single implants and buccal bone grafts in the anterior maxilla: measurements of buccal crestal contours in a 6-year prospective clinical study. AB - BACKGROUND: Patients provided with buccal bone grafts seem to lose a substantial part of the graft in the short term. PURPOSE: To measure long-term changes in buccal and proximal tissue volumes after local bone grafting and single implant treatment. MATERIALS AND METHODS: Eight of 10 originally treated male patients were followed up for 6 years after treatment with buccal bone grafts in the central incisor region. After a healing time of 6 months, a two-stage implant surgery procedure was performed followed by single crown placement. Clinical photographs and impressions were taken prior to the surgical interventions and after crown placement and at first and fifth annual checkups. The photographs were analyzed with regard to papilla regeneration by means of a clinical papilla index. The models were used to measure the clinical length of teeth and tooth movements adjacent to the implants. Changes in buccal crest volume during the study period were measured by means of optical scanning of obtained study models. RESULTS: Papillae volume increased significantly (p<.05) during the first year, thereafter showing a slow further increase during the 4 following years. Three of the patients (38%) presented small movements of their adjacent central incisor in a vertical or palatal direction of less than 1 mm during the follow-up period. All patients showed resorption during the first year after grafting (p<.01), in which three patients (38%) had lost basically all of increased volume at second surgery. After abutment or crown placement, all patients showed an increased volume (p<.01), followed by an average reduction during the first year, reaching a significant level in the apical part of the crest (p<.05). Thereafter, a relatively stable average situation was observed during the following 4 years, with individual variations, however. CONCLUSION: Local bone grafting seems to create sufficient bone volume for implant placement after 6 months, but individual variations in resorption pattern make the grafting procedure unpredictable for long-term prognosis. Instead, the abutment and the crown seem to play a more important role for building up and maintaining the buccal contour in the coronal part of the crest long term. PMID- 16219243 TI - Implant stability during initiation and resolution of experimental periimplantitis: an experimental study in the dog. AB - BACKGROUND: Histologic studies have demonstrated the possibility to reestablish direct bone-implant contacts after ligature-induced periimplantitis. The influence of the reosseointegration on the stability of implants is not known. PURPOSE: The aim of the present investigation was to study bone tissue and associated implant stability alterations that occurred during induction and resolution of periimplantitis using resonance frequency analysis (RFA), radiography, and histology. MATERIALS AND METHODS: Three implants with smooth (turned) or roughened (SLA) surfaces were placed in each side of the edentulous mandible of four dogs. Experimental periimplantitis was induced for 3 months. Five weeks later, the animals were treated with antibiotics and surgical therapy and were followed for another 6 months. Periapical radiographs and RFA were used to evaluate marginal bone levels and implant stability throughout the study period. After termination, the tissue-implant interface was evaluated by light microscopy in ground sections. RESULTS: There was a linear relationship between radiographic and RFA findings because continuous loss of marginal bone and a decrease in implant stability were observed for both implant surfaces during the periimplantitis period. Antibiotic treatment and surgical therapy resulted in some reosseointegration, which was more marked for the SLA surface. The resonance frequency values corresponded well to the histometric measurements because reosseointegration resulted in an increase in implant stability. CONCLUSIONS: The findings from the present study indicate a linear relationship between marginal bone level and resonance frequency value. It is suggested that the RFA technique is sensitive and may be used to detect even a minor change in the level of bone implant contact. PMID- 16219244 TI - Screw preloads and measurements of surface roughness in screw joints: an in vitro study on implant frameworks. AB - BACKGROUND: With the development of milled titanium implant frameworks, new surfaces that have not previously been studied are now being used in screw joints. PURPOSE: The aims of the present study were to compare the preload produced in screw-retained titanium and gold alloy frameworks and the preload for titanium frameworks before and after the application of veneers. Another aim was to try to relate the surface roughness of the screw joints to variations in preload. MATERIALS AND METHODS: Ten identical titanium and five gold alloy frameworks were fabricated. The gold screws were tightened to 10 Ncm. Preload measurements were made for the gold alloy frameworks and before and after the porcelain or acrylic resin veneers had been applied to the titanium frameworks. Surface roughness measurements were made after preload measurements on the screw joint surfaces of the titanium frameworks and corresponding gold screws. RESULTS: The preloads for the titanium and gold alloy frameworks were similar. Preload in both types of frameworks decreased after repeated torques (p<.05-.01) but was unaffected by the application of veneering materials to the titanium frameworks (p>.05). No relationship (p>.05) between preload and surface roughness characteristics was observed. Loaded titanium framework screw sites, however, had lower mean S(a) values than unloaded sites (p<.001), whereas the surfaces of loaded gold screws had higher mean S(a) values compared with the surfaces of control gold screws (p<.05-.001). CONCLUSION: When using gold screws, milled titanium frameworks have preloads similar to those of gold alloy frameworks and preloads for both decrease after repeated tightening. The preload was similar before and after the veneering of the titanium frameworks. Unloaded milled titanium screw sites had rougher surfaces than loaded, and loaded gold screws had rougher surfaces than unloaded. However, no correlation between screw joint surface and preload was observed for veneered titanium frameworks. PMID- 16219245 TI - Biologic response of immediately versus delayed loaded implants supporting ill fitting prostheses: an animal study. AB - BACKGROUND: Computer-assisted preoperative implant planning and transfer toward the patient allow the production of a prosthesis prior to surgery. This implies that the prosthesis can be installed immediately following implant insertion. An inherent disadvantage of this is a cumulated error, which can lead to prosthesis misfit owing to topographic deviations of the planned versus the installed implants. PURPOSE: The aim of this study was to determine whether prosthesis misfit is compromising the osseointegration of immediately versus delayed loaded implants and whether freshly installed implants adapt to the prosthesis. MATERIALS AND METHODS: In each of five New Zealand White rabbits, two experimental conditions were compared. One tibia harbored the so-called test implant, which originally showed a vertical misfit of about 500 microm with the prosthesis to which it was tightened immediately after implant installation. The control implant was installed in the other tibia and was allowed to heal during 9 weeks before the prosthesis with the vertical misfit of about 500 microm was connected to it. The prostheses were left in place for 12 weeks, after which the animals were sacrificed. RESULTS: All implants healed uneventfully. There were no statistically significant differences between the biologic responses of test and control implants. With a three-dimensional laser scanner, significantly more displacement of the test implants toward the prostheses was observed compared with the control implants. This led to a significant decrease in prosthesis misfit for the test implants compared with the control implants. CONCLUSIONS: This study indicates that prosthesis misfit does not per se lead to biologic failure of immediately loaded or of already osseointegrated implants. In addition, immediately loaded implants seem to topographically adapt to the prosthesis, thereby minimizing the existing misfit. PMID- 16219247 TI - A retrospective evaluation of treatments with implant-supported maxillary overdentures. AB - BACKGROUND: Considerably lower success rates have been presented for implant supported overdentures in the maxilla compared with the mandible. PURPOSE: The aim of the study was to report the outcome of implant-supported maxillary overdentures from one clinic. METHODS: All patients treated with implant supported maxillary overdentures in the Department of Prosthetic Dentistry, Central Hospital, Skovde, Sweden, between 1993 and 2002 were identified from patient charts and included in the study. All patients had a rigid cast gold alloy bar designed with ball attachments retaining an overdenture. RESULTS: Twenty-seven subjects were included, of whom 13 were originally planned for overdenture treatment (group 1) and the other 14 for a fixed prosthesis (group 2). The mean observation period was 5.7 years for subjects in group 1 and 5.5 years for those in group 2. One hundred forty-five implants were placed, and the majority of the failures were diagnosed as early ones and were found in group 2. The cumulative implant survival rate after 5 years was 77% in group 1 and 46% in group 2. The probability of having implant failure was almost three times higher among subjects in group 2 compared with subjects in group 1. Most technical and biologic complications were related to the retention system. CONCLUSION: Maxillary implant-supported overdentures show a high implant failure rate, but fewer implant failures occurred for patients originally planned for overdenture treatment. PMID- 16219246 TI - Evaluation of 31 zygomatic implants and 74 regular dental implants used in 16 patients for prosthetic reconstruction of the atrophic maxilla with cross-arch fixed bridges. AB - BACKGROUND: The use of a specially designed implant to be anchored in the zygomatic body has been proposed as an alternative to bone grafting in the prosthetic rehabilitation of the severely resorbed maxilla. However, few studies have evaluated the long-term stability and soft tissue conditions of zygomatic implants. PURPOSE: The aim of this retrospective study was to evaluate the clinical performance of zygomatic implants when used for prosthetic reconstruction of atrophic maxillae. MATERIALS AND METHODS: Sixteen patients consecutively treated with 31 zygomatic implants and 74 additional dental implants from 1998 to 2002 were retrospectively evaluated and prospectively followed using a standardized clinical and radiographic study design. Data were collected from the time of implant treatment until the last follow-up RESULTS: The follow-up period ranged from 9 to 69 months from the day of implant treatment, with a mean of 46.4 months (3 years, 10 months). Three (9.7%) of the 31 zygomatic implants were surgically removed because of recurrent sinusitis. Three (4.1%) of the 71 additional dental implants failed to integrate. Poor oral hygiene and gingivitis were seen at most zygomatic implant sites (10/16). Local infections were observed in 9 of 16 patients. Sinusitis occurred in 6 patients. All patients (16/16) eventually received fixed bridges, which were stable throughout the observation period. CONCLUSIONS: The results showed an acceptable outcome with regard to implant and prosthetic survival rates. However, postoperative complications not related to implant and prosthesis stability were frequent. Further investigations of the long-term performance of zygomatic implants and with a focus on soft tissue and maxillary sinus health are needed. PMID- 16219248 TI - Detection and measurements of soluble intercellular adhesion molecules at implants and teeth: a comparative study. AB - BACKGROUND: Adhesion molecules on endothelial cells and in the periodontal tissues control the immigration and retention of cells. The level of soluble intercellular adhesion molecules (sICAMs) has been used as a marker of the severity and/or extent of the inflammatory process in a wide range of pathologies, including periodontitis. PURPOSE: This study was designed to detect and compare sICAM-1 at teeth and implants in relation to clinical periodontal and periimplant parameters. METHOD: Regular recall patients with (1) implants and teeth, (2) implants, and (3) teeth were examined. Samples of sulcus fluid were collected from teeth and implants. The concentration of sICAM-1 was measured by enzyme-linked immunoabsorbent assay. Periodontal parameters were recorded after sampling. RESULTS: The range of measured sICAM-1 was large (from below 100 to 1,200 ng/mL). The concentration of sICAM-1 was not different for teeth and implants but was significantly elevated in sites with positive bleeding on probing (BoP), namely, 571 ng/mL at teeth and 529 ng/mL at implants compared with 150 ng/mL and 169 ng/mL, respectively, with negative BoP. The regression analysis showed that the concentration of sICAM-1 was highly associated with positive BoP but was not dependent on the fluid volume. CONCLUSIONS: A similarity of the sulcus fluid at teeth and implants was observed with regard to the detection of sICAM-1. PMID- 16219249 TI - [Update in pediatric rheumatology]. PMID- 16219250 TI - [Neonatal morbidity and mortality according to intrauterine growth pattern]. AB - OBJECTIVE: To evaluate the influence of intrauterine growth on neonatal morbidity and mortality in live neonates born in a regional tertiary care center. PATIENTS AND METHODS: A retrospective cohort study was performed. Data from 12,311 live neonates (LN) born in our hospital (from January 1999 to December 2003) were reviewed to analyze neonatal morbidity and mortality according to whether the LN were appropriate (AGA), small (SGA) or large (LGA) for gestational age. The variables collected from birth to hospital discharge were: gestational abnormalities, gender, delivery type, Apgar score, respiratory distress, presentation of obstetric trauma (including cephalohematomas), hypoglycemia, hypocalcemia, hyperbilirubinemia, congenital malformations, heart disease, and type of hospital discharge. Differences were examined among groups. We used the chi-squared test to compare the frequency of these variables in each group. RESULTS: Data from 12,311 LN were entered into the database; 11,182 (90.8%) were AGA, 743 (6.1%) were LGA and 386 (3.1%) were SGA. There were 52.58% boys and 47.42% girls, and 5.5% of LN had macrosomia (birth weight > 4,000 g). A total of 0.39% of LN died before hospital discharge. Among LN, there were 1,215 preterm infants, representing 9.89% of all LN and 2.63% died (SGA 25% and AGA 75%). There was a prevalence of boys in the LGA group and of girls in the SGA group (p = 0.000). The presence of maternal diabetes (pregestational or gestational) was significantly higher (p = 0.000) in the LGA group. Maternal hypertension, smoking and drug addiction were significantly higher in the SGA group. The finding of obstetric trauma was significantly higher in the LGA group (p = 0.000). The proportion of congenital malformations, hypocalcemia and hypoglycemia was higher in the SGA group than in the AGA and LGA groups (p = 0.000). Neonatal mortality was significantly higher (p = 0.000) in the SGA group and preterm infants. CONCLUSIONS: Neonates with deviations in the pattern of intra-uterine growth had worse outcome. Prognosis was worse in SGA neonates, followed by LGA neonates, than in AGA neonates. PMID- 16219251 TI - [Markers of poor prognosis in convulsive status epilepticus]. AB - OBJECTIVE: To characterize the variables that might be associated with mortality and the development of neurological deficits in children with convulsive status epilepticus. PATIENTS AND METHODS: Children older than 1 month and younger than 15 years who were admitted to the pediatric intensive care unit of a university hospital between 2001 and 2004 were reviewed. Epidemiologic and clinical factors that might be related to neurological outcome at discharge from the unit were analyzed. RESULTS: Forty-one patients (median age 24 months) were included. A total of 48.3% developed refractory convulsive status epilepticus. Six patients died (mortality 14.6%) during their intensive care unit stay and neurologic worsening was observed in 8.6% of survivors (adverse outcome in 22%). Symptomatic epilepsy was present in all patients who died and in 88.9% of those who recovered with severe neurologic sequelae. Uni- and multivariate analysis showed that adverse outcome was related to symptomatic origin and refractory convulsive status epilepticus (p < 0.05). CONCLUSIONS: Mortality and morbidity is high in childhood convulsive status epilepticus. Refractory convulsive status and symptomatic origin were markers of poor outcome. Children who did not have symptomatic epilepsy had a favorable outcome. PMID- 16219252 TI - [Telephone triage performed by emergency room physicians]. AB - OBJECTIVE: To determine the applicability and utility of a telephone triage performed by physicians in a pediatric emergency department (PED). PATIENTS AND METHOD: All the telephone consultations registered between 2003-10-1 and 2004-10 3 were reviewed. The variables analyzed were: telephone call record (n = 2,560), compliance with the advice given (randomized sample of patients who were not referred to the PED and all patients who were referred) and parent satisfaction (callers over a 1-month period were contacted). A protocol for answering queries was organized and residents received specific training. RESULTS: There were 59,088 episodes and 2,560 calls were registered (1 call/23 episodes). Forty-eight percent of the calls were received between 5 and 11 pm. All calls were answered by a physician (72% residents, 28% attending physicians). The most frequent reason for calling was to seek advice on symptoms. Eighty-two percent of the calls were resolved through telephone instructions to be carried out in the home. A total of 274 patients were advised to attend the PED, but 29% did not attend, usually because the child's symptoms improved. Twenty patients were admitted to the hospital (6 to wards, 14 to the observation unit). Of the patients not advised to attend, 21% attended the PED and 0.9% were admitted (compared with 7.2% in the referral group, p = 0.0001). More than 90% of the families questioned were satisfied with the advice given. Seventy-five percent would have attended the PED if telephone consultation had not been available. By giving telephone advice, we avoided 115 visits in 1 month. COMMENTS: If special training programs and answering systems are established after a training period, telephone consultation in a PED is a safe and useful method of performing patient triage. Satisfaction among families was high. PMID- 16219253 TI - [Systemic lupus erythematosus in children]. AB - Pediatric systemic lupus erythematosus (pSLE) is a chronic mutisystemic autoimmune disease with complex clinical manifestations. Although the presentation, clinical manifestations, immunological findings and treatment issues of pSLE are similar to those of adult SLE patients, there are special issues which need to be considered when dealing with SLE in children. During the last decade survival has improved remarkably as a result of earlier diagnosis, recognition of milder disease and better approaches to therapy. However, pSLE remains a potentially serious condition. Although the pathogenesis of SLE remains poorly understood, susceptibility involves a combination of environmental, hormonal and genetic factors. Better understanding of SLE pathogenesis will hopefully lead to more specific and less toxic therapies for this disease. PMID- 16219254 TI - [Syncope in adolescents. Diagnostic and therapeutic approach]. AB - Syncopal episodes are frequent in adolescence. Syncope is usually a benign, self limiting condition but it may be a warning sign of serious disease that must be diagnosed and appropriately treated. The present article provides a review of the basic principles of the differential diagnosis of syncope in the adolescent patient and treatment recommendations. PMID- 16219255 TI - [Breastfeeding. How to promote and support breastfeeding in pediatric practice. Recommendations of the Breastfeeding Committee of the Spanish Association of Pediatrics]. PMID- 16219256 TI - [Recommendations for the prevention of respiratory syncytial virus infection]. AB - Premature babies < or = 35 weeks gestation, with or without chronic lung disease (CLD), and infants affected by severe congenital heart disease should be considered high risk population for RSV infection and hospitalization. Hygienic measures and RSV monoclonal antibodies (palivizumab) have been found useful in decreasing rates of RSV hospitalization in these patients. Guidelines for their administration include: a) strongly recommended use in premature babies < or = 28 weeks gestation, or born between 29-32 weeks gestation and less than 6 months at start of RSV station or discharged along it, or affected by CLD in treatment during last 6 months or by severe congenital heart disease. Last two groups could be prophylaxed for two RSV seasons. b) Recommended use among premature babies between 32-35 weeks gestation and less than 6 months of age and presenting two or more risk factors: chronologic age < 10 weeks at start of RSV station, breastfeeding < or = 2 months (physician prescription), sibling < 14 years old, day-care assistance, family history of wheezing, > or = 4 adults at home, airways malformation or neuromuscular disease. PMID- 16219257 TI - [Topiramate and pregnancy. Neonate with bone anomalies]. AB - Topiramate is a new generation, antiepileptic drug used for the treatment of persistent partial crises. To date no specific teratogenic effects have been reported in humans, but they have appeared in experimental animals. We present the case of a neonate whose mother suffered from partial epilepsy, which was treated with topiramate throughout pregnancy at doses of 300 mg per day. When the child was born agenesis of the right thumb, hypoplasia of the left thumb, and syndactylia of the second and third toes of the foot with agnesis of some phalanges, and hypoplasia of the right orbicular muscle in the mouth were observed. No etiologic cause was found. We discuss whether there could have been a causal relationship with topiramate monotherapy. PMID- 16219258 TI - [Asperger syndrome associated with macrosomia and sexual identity disorder]. AB - Asperger Syndrome is a pervasive developmental disorder of unknown origin, characterized by pedantic language, lack of reciprocity in social interactions, unusual interests, motor clumsiness and normal or above average intelligence quotient, among other symptoms. Since 1994 it has been defined as a specific entity. We describe the case of a boy with this syndrome, with elevated body weight and height and sexual identity disorder. These alterations have not previously been described in the scientific literature on Asperger syndrome. PMID- 16219259 TI - [Rotavirus diarrhea in a population covered by private health insurance in Cordoba, Argentina]. AB - BACKGROUND: Rotavirus is the most common cause of severe diarrhea worldwide, especially in children under 5 years of age. OBJECTIVES: To study the incidence of rotavirus diarrhea in a population in Cordoba, Argentina. To determine signs and symptoms with potential predictive clinical diagnostic value and evaluate how rotavirus diarrhea affects length of hospital stay. MATERIAL AND METHODS: A case control study was performed from April 1, 2002 to March 31, 2003. RESULTS: Seventy-three patients were included. There were 33 rotavirus (1) cases and 40 rotavirus (-) controls. The mean age of the rotavirus (1) patients was 16.8 6 2.3 months. There were 14 (42.4 %; 95 % CI: 27.2-59.2) girls and 19 (57.6 %; 95 % CI: 40.8-72.8) boys. The seasonal peak of the disease was observed in the fall (30.3 %; 95 % CI: 17.4-47.3) and at the beginning of the austral winter (39.3 %; 95 % CI: 24.7-56.3). The signs and symptoms associated with the infection were vomiting (OR 8.40; 95 % CI: 2.39-31.33) and dehydration (OR 3.73; 95 % CI: 1.06 13.81). In the 33 rotavirus (1) patients, the mean length of hospital stay was 2.6 days. CONCLUSIONS: One half of the hospitalized cases were produced by rotavirus. The seasonal peak was observed in autumn and winter (southern hemisphere). Rotavirus (1) patients more frequently presented severe vomiting and dehydration than rotavirus (-) patients, although length of hospital stay and diarrhea were longer in the control group. PMID- 16219260 TI - [Systemic involvement in premature infants after intranasal administration of tetryzoline drops]. PMID- 16219261 TI - [Recurrent neonatal adrenal cysts associated with hemihypertrophy]. PMID- 16219262 TI - [Recurrence of Salmonella meningitis after cefotaxime therapy]. PMID- 16219263 TI - [Linear porokeratosis of Mibelli]. PMID- 16219264 TI - [Pilous dysplasia: a new case of monilethrix]. PMID- 16219265 TI - [ATVs represent a significant health risk to children and teenagers]. PMID- 16219266 TI - [Frequent errors in the comparison of results of studies on burn-out]. PMID- 16219267 TI - [Clinical and pharmacological databases for personal digital assistants]. PMID- 16219269 TI - [Epilepsy and hair alterations in a neonate]. PMID- 16219270 TI - [Early detection of hypacusia in neonates]. PMID- 16219271 TI - [Epidemiological trends in new diagnoses of HIV-1 infection in children]. AB - INTRODUCTION: In the last few years, the immigrant population in Spain, as well as the number of HIV-infected patients from countries where the prevalence of HIV 1 infection is higher, have continued to increase. No data are available on the impact of immigration on new cases of HIV-1 diagnosed in children. The objective of this study was to assess the clinical characteristics and determine the epidemiological trend of new diagnoses of pediatric HIV infection in Madrid in the last 8 years. PATIENTS AND METHODS: We performed a retrospective study through review of the medical records of children in whom a new diagnosis HIV-1 infection was made in nine hospitals in the Autonomous Community of Madrid from 1997 to 2004. Two periods were compared: before December 2000 (period A) and after December 2000 (period B). Children born outside Spain or those whose parents had arrived in Spain in the previous 10 years were considered immigrants. RESULTS: During the period analyzed, 97 children with HIV infection were identified and transmission was vertical in 93. Overall, 33 (34%) were immigrants (51% from Latin America, 27% from sub-Saharan Africa, 15% from the Maghreb, and 6% from Eastern Europe). The percentage of immigrant children increased from 26% (19/72) in period A to 56% (14/25) in period B (p: 0.013). The mean age at diagnosis was 3.9 +/- 0.8 years in the immigrant group and 3.7 +/- 0.5 in the group of Spanish children (p > 0.05). No differences were found in the type of transmission, CDC-clinical class, viral load or CD4 cells between Spanish and immigrant children. CONCLUSIONS: The proportion of immigrant children with HIV-1 infection is increasing in Madrid. In our experience, we found no significant differences in clinical or immunological category between immigrant and Spanish children with regard to diagnosis of HIV infection. PMID- 16219272 TI - [Acute disseminated encephalomyelitis. Experience of a tertiary hospital in Spain]. AB - INTRODUCTION: To describe the epidemiological, clinical, microbiological, neuroimaging and laboratory features, treatment, and outcome in a cohort of children with acute disseminated encephalomyelitis (ADEM). PATIENTS AND METHODS: Retrospective chart review was performed of children with a diagnosis of ADEM over a 23-year period in a tertiary hospital in Spain. RESULTS: Twelve cases were identified. Ten cases (83%) occurred after 1992. Nine patients (75%) presented between April and September. The mean age was 6 years. Nine patients (75%) were male. Fifty percent of the patients had a history of infectious disease or vaccination. The most frequent nonspecific symptom was fever in 75%. The most frequent neurological manifestations were motor deficits and altered consciousness in 75%. Cerebrospinal fluid abnormalities were found in 83%. All patients had at least one brain scan and one brain magnetic resonance imaging (MRI) scan. Three patients underwent spinal MRI. The sensitivity of MRI was greater than that of the scanner in the diagnosis of ADEM. An etiologic diagnosis was made in four patients: Mycoplasma pneumoniae, beta hemolytic streptococcus group A, Epstein-Barr virus and measles-mumps-rubella vaccination. Eleven patients were treated with corticosteroids and one was treated with intravenous immunoglobulin therapy. One patient died while 75 % of the patients had a good outcome. CONCLUSIONS: ADEM is in an infrequent disease in children. The clinical features are similar to those of infectious encephalitis. Etiologic diagnosis is difficult to establish but this entity is usually preceded by an infection. The neuroimaging test of choice to establish the diagnosis is MRI. In most patients, the prognosis is good. PMID- 16219273 TI - [Ibuprofen versus indomethacin in the treatment of patent ductus arteriosus in preterm infants]. AB - INTRODUCTION: Persistent patent ductus arteriosus (PDA) is a common entity in preterm infants. The most commonly used pharmacological treatment to close the ductus is indomethacin but it can affect cerebral, renal and mesenteric blood flow. Ibuprofen has recently been shown to be effective in closing PDA with fewer hemodynamic effects. In this study we compared the safety and efficacy of ibuprofen and indomethacin in the treatment of PDA in preterm infants. MATERIAL AND METHODS: A randomized trial was performed. Premature infants with symptomatic PDA confirmed by echocardiography in the first week of life and who required respiratory support were included. The patients were randomly assigned to receive either intravenous indomethacin or ibuprofen. The rate of ductal closure, need for additional treatment, complications, and clinical course were evaluated. RESULTS: Twenty-four patients were treated with indomethacin and 23 with ibuprofen. The clinical characteristics before treatment were similar in both groups. Both treatments were effective in closing PDA (87.5% in the indomethacin group and 82.6% in the ibuprofen group). The two cohorts did not differ in the rate of reopening, need for a second pharmacologic treatment, or surgical ductal ligation. No patient in the ibuprofen group developed gastrointestinal adverse effects, but two infants in the indomethacin group had isolated bowel perforation and one had necrotizing enterocolitis. Transient renal dysfunction developed in seven patients (29%) in the indomethacin group versus two (9%) in the ibuprofen group. Transient renal insufficiency was found in one patient in the indomethacin group and in none in the ibuprofen group. The rate of other complications was similar in both groups. CONCLUSIONS: In our trial ibuprofen was as effective as indomethacin in closing PDA. No significant differences were found in the incidence of complications but fewer renal complications and no gastrointestinal complications were found in the ibuprofen group. PMID- 16219274 TI - [Study of bone mass in patients with growth hormone deficiency]. AB - OBJECTIVES: To evaluate bone mineral density by radiogrametric study of metacarpal bone diameter and cortical thickness in patients with growth hormone deficiency (GHD) before and during treatment with growth hormone (GH). PATIENTS AND METHODS: We studied 92 children with GHD (60 boys and 32 girls) divided into two groups: group I: 66 previously untreated patients (42 boys and 24 girls) aged between 3 and 14 years old; group II: 66 patients (42 girls and 24 boys) treated with GH and with a mean age of 10.2 +/- 3.1 years at treatment onset. Bone mass was studied indirectly by radiogrametry; the bone diameter and cortical thickness of the 2nd-3rd and 4th metacarpal bones were measured with a magnifying glass. As reference standards we used the Spanish longitudinal growth and development study (Andrea Prader Center, Zaragoza) in children aged between 0.5 and 9 years and the Swiss longitudinal standards in children aged 10 years of age and older. Statistical significance was set at p < 0.05. RESULTS: Group I (spontaneous evolution): cortical thickness values were below the mean with statistically significant differences al 11, 12 and 13 years of age in girls and at 12, 13 and 14 years in boys. Bone diameter was diminished compared with controls in all the study periods and was significantly reduced at 8, 9, 10 and 11 years of age in girls and at 8, 10, 11, 12, 13 and 14 years in boys. Group II: (effect of GH treatment): cortical regression analysis showed a sharp increase in the first year of treatment with a subsequent moderate increase, which was statistically significant. Bone diameter showed a similar pattern with a significant increase which was more pronounced in the first period. CONCLUSIONS: Children with GHD have decreased bone mass before initiation of treatment and therefore show deficient acquisition of peak bone mass, which in normal conditions occurs during in the first 4-5 years of life and during adolescence. GH replacement therapy leads to recovery of bone mass, which is more pronounced in the first year of treatment and prevents the progressive reduction that appears in untreated patients. Therefore, GH treatment plays an important role in peak bone mass acquisition in children with GHD. PMID- 16219275 TI - [Bone mineral turnover and bone densitometry in patients with a high-risk diet: hyperphenylalaninemia and galactosemia]. AB - INTRODUCTION: Patients with any type of congenital metabolism error are at risk for developing osteoporosis. To gain further insight into the physiopathology of this disease, we studied bone mineral turnover in 10 children with hyperphenylalaninemia, seven with phenylketonuria and six with galactosemia. Oral intake was strictly controlled and the children followed recommendations for physical exercise. MATERIAL AND METHOD: Markers of bone resorption (hydroxyproline and pyridinoline in urine samples) and markers of bone formation (levels of osteocalcin and C-terminal procollagen peptide type I) were analyzed. Bone mineral density was analyzed by ultrasound densitometry. RESULTS: A non significant reduction in bone densitometry with respect to the normal population was observed. Bone mineral turnover was slightly diminished in patients with phenylketonuria but was within the normal range in patients with hyperphenylalaninemia and galactosemia. CONCLUSION: Adequate control of dietary intake of both proteins and minerals, as well as a healthy lifestyle, can prevent the development of significant alterations in bone mineralization. PMID- 16219276 TI - [Evaluation of a universal screening program for hypacusia in neonates]. PMID- 16219278 TI - [Use of somatostatin in five neonates with chylothorax]. AB - BACKGROUND: The addition of somatostatin to the conventional treatment of neonatal chylothorax has been described in isolated cases. OBJECTIVE: To describe the results obtained in a series of five patients with neonatal chylothorax treated with somatostatin. PATIENTS: Five neonates (gestational age range: 29-39 weeks) diagnosed with chylothorax of various etiologies were included. Chylothorax was congenital in two neonates, secondary to congenital diaphragmatic hernia repair in two neonates and secondary to thrombosis in the superior vein cava in one neonate. All the neonates were started on conservative therapy and intravenous somatostatin in distinct doses ranging from a bolus of 2 microg/kg/12 h to continuous perfusion at 10 microg/kg/h. RESULTS: In all patients the chylous drainage was stopped. No adverse effects were observed. CONCLUSIONS: Somatostatin can be a safe and effective option in the treatment of both primary and secondary neonatal chylothorax refractory to conservative treatment. PMID- 16219277 TI - [Clustering of cardiovascular risk factors in obese offspring of parents with essential hypertension]. AB - INTRODUCTION: The prevalence of obesity in industrialized countries is increasing and is closely related to essential hypertension (EHT) in adolescents. OBJECTIVE: To analyze the prevalence of obesity and its association with other known cardiovascular risk factors in a sample of children and young adults with at least one parent with EHT. METHODS: The EHT group consisted 51 children and young adults (28 males [aged 5.4-25.6 years]) with at least one parent with EHT. The control group comprised 73 healthy normotensive children and young adults (43 males [aged 7.2-25.2 years]) who completed the follow-up visits of the RICARDIN study. Blood pressure (BP) was measured with a standardized technique using a mercury sphygmomanometer. A 12-hour fasting blood sample was taken for lipid profile and high sensitivity C-reactive protein (CRP) determinations. Financial support: FIS 03/0350, ESV Foundation Grant, 2003. RESULTS: The prevalence of obesity was five times higher in the EHT group than in controls (19.6% vs. 4.1%, p = 0.007). In this group, obese subjects showed higher systolic BP (122.0 vs. 110.4 mmHg p = 0.004) and lower high-density lipoprotein cholesterol (HDL-C) levels (47.6 vs. 58.0 mg/dl, p < 0.05). After adjustment for age and systolic BP, obese subjects in the EHT group showed significantly higher CRP values than non obese subjects in this group (p = 0.024). CONCLUSIONS: The prevalence of obesity is higher in the offspring of parents with EHT than in non HT-prone subjects. Clustering of other additional risk factors indicates the need for high-risk preventive interventions in this group of children and young adults. PMID- 16219279 TI - [Disseminated histoplasmosis]. AB - We describe two cases of disseminated histoplasmosis, which are of interest due to their severity and the infrequency of this infection in our environment. Both children were immunocompromised immigrants from Latin America who developed prolonged fever, weight loss, hepatosplenomegaly and pancytopenia. One patient had respiratory symptoms with associated alterations on x-ray, while no radiological alterations were found in the other patient. Despite administration of broad-spectrum antibiotics and extraction of samples for microbiological analysis, both patients had a rapid and fatal outcome and the diagnosis was made post mortem. Because of its severity, disseminated histoplasmosis should be considered in the differential diagnosis of fever, hepatosplenomegaly and pancytopenia, with or without alterations on chest x-ray, in immunosuppressed children who were born in or have visited endemic regions. In these patients, therapy should be started immediately without waiting for the results of diagnostic tests. PMID- 16219280 TI - [Brain abscess in children]. AB - INTRODUCTION: Brain abscess is a rare infection in infants. Morbidity and mortality are high but have decreased due to advances in neuroimaging studies and the use of new antibiotics. We describe six cases of brain abscess diagnosed at the Gregorio Maranon Children's Hospital between January 1996 and September 2003. METHODS: We performed a retrospective chart review of patients with brain abscess. The variables analyzed were age, sex, clinical symptoms and signs, radiological studies, etiology, therapy, and clinical outcome. RESULTS: Age ranged from 8 to 15 years (mean age: 11 years). There were three girls and three boys. The most frequent symptoms were neurological with associated sinusitis in four patients, congenital cyanogenic cardiopathy in one patient and meningitis in one patient. Diagnosis was established through computed tomography (CT) of the brain in five patients and through magnetic resonance imaging (MRI) in one patient. In five patients the abscess was located in the frontal lobe. All patients received broad-spectrum antibiotics and five underwent surgical drainage. Two patients had neurological sequelae. CONCLUSIONS: Although rare, brain abscess should be considered in patients with neurological manifestations associated with otorhinolaryngological infections or congenital cyanotic cardiopathy. When suspected, a CT or MRI must be performed to rule out this diagnosis and, if confirmed, prompt therapy with broad-spectrum antibiotics should be started. Surgical drainage may be needed. PMID- 16219281 TI - [Graves' disease in preschool children]. AB - Graves' disease, which is the main cause of hyperthyroidism in the pediatric age group, is very rare in pre-school children. We describe the cases of four girls, aged less than 6 years old, out of a total of 30 patients diagnosed with Graves' disease between 1985 and 2004. Investigations were motivated by tachycardia, chronic diarrhea, language development delay or thyroid nodules detected by cervical ultrasonography. In three of the four patients height and bone age were advanced. In all patients goiter was small or absent. None of the patients had ophthalmic disease. In all patients free T3 and T4 were elevated and thyroid stimulating hormone was suppressed. Three patients were positive for thyroid stimulating immunoglobulins (the method was not available for the oldest case). Two patients showed complete resolution after 5 years of treatment with carbimazole. The remaining two patients are still under treatment and no adverse effects have been documented. PMID- 16219282 TI - [Liver abscess. An uncommon complication of acute appendicitis]. PMID- 16219283 TI - [Abdominal lymphangioma infected by Salmonella enteritidis]. PMID- 16219284 TI - [Treatment of lymphadenitis]. PMID- 16219285 TI - [Gastrointestinal bleeding due to vascular anomaly]. PMID- 16219287 TI - [Asymmetric periflexural exanthema: apropos of a case]. PMID- 16219288 TI - [Atloaxoid spondylitis. An infrequent cause of torticollis]. PMID- 16219289 TI - [Rothmund Thomson syndrome]. PMID- 16219290 TI - [Maculopapulous exanthema with particular distribution]. PMID- 16219291 TI - Relationship of the unstimulated whole saliva flow rate and salivary gland size estimated by magnetic resonance image in healthy young humans. AB - OBJECTIVE: The aim of this study was to examine the relationships between gland sizes and the flow rate and composition of the unstimulated whole saliva in humans. DESIGN: In 28 healthy young adults, the sizes of the three major salivary glands were estimated by use of a magnetic resonance (MR) imaging technique. Unstimulated whole saliva was collected for 5 min by the spitting method, and the flow rate and the concentrations of total protein, Na(+) and K(+) and pH were measured. RESULTS: The estimated sizes of the parotid and submandibular glands showed a significant positive correlation with the flow rate and the secretion rate of total protein in the unstimulated whole saliva, but that of the sublingual glands did not. Concerning the concentrations of Na(+) and K(+) and pH, there were no correlations with the salivary gland sizes. CONCLUSIONS: The results suggest that the larger the sizes of the parotid and submandibular glands, the faster the fluid flow and protein secretion rates in unstimulated whole saliva. PMID- 16219292 TI - INI1/hSNF5/BAF47 represses c-fos transcription via a histone deacetylase dependent manner. AB - INI1/hSNF5/BAF47 is a core component of the hSWI/SNF ATP-dependent chromatin remodeling complex. It has been suggested that INI1/hSNF5/BAF47 contributes to the regulation of many genes. In this report, we showed that the overexpression of INI1/hSNF5/BAF47 repressed c-fos promoter activity and endogenous c-fos transcription in 293T cells, and the siRNA targeting INI1/hSNF5/BAF47 (siINI1) reversed the inhibitory effect. Histone deacetylation by histone deacetylases (HDACs) was necessary for the repression of c-fos transcription by INI1/hSNF5/BAF47. HDAC and INI1/hSNF5/BAF47 functioned together to suppress c-fos transcription. ChIP experiments demonstrated that INI1/hSNF5/BAF47 could be recruited to the region of c-fos promoter to reduce histone acetylation. Altogether, these data show that INI1/hSNF5/BAF47 represses c-fos transcription via a histone deacetylase (HDAC)-dependent manner. PMID- 16219293 TI - Mutational study of human phosphohistidine phosphatase: effect on enzymatic activity. AB - Although protein histidine phosphorylation is estimated to account for about 6% of total protein phosphorylation in eukaryotes, knowledge on histidine phosphorylation and dephosphorylation is still limited. Recently, a few reports have appeared on a mammalian 14-kDa phosphohistidine phosphatase, also named protein histidine phosphatase. Molecular cloning of the protein has opened possibilities for exploring its properties and physiological role. In the present work, we have searched for potential active site residues in the human phosphohistidine phosphatase by point mutations of conserved histidine and arginine residues to alanine. When assayed by the phosphohistidine-containing peptide succinyl-Ala-His(P)-Pro-Phe-p-nitroanilide, mutants H53A and H102A showed no detectable activity. Compared to the wild-type recombinant enzyme, the specific activity of mutant R45A was decreased by one order of magnitude, that of mutant R78A was decreased by about 30%, while that of mutant H81A was essentially unchanged. These results will facilitate future studies of the reaction mechanism, substrate binding, and molecular structure of the phosphohistidine phosphatase. PMID- 16219294 TI - High level production of secreted proteins: example of the human tissue inhibitor of metalloproteinases 1. AB - The major difficulty for high-throughput screening of therapeutic protein candidates in experimental animal models of pathologies or for structural studies is their fast and efficient production. The tissue inhibitors of metalloproteinases (TIMPs) considered to play a role in many physiological and pathological processes, such as arthritis or cancer, by inhibiting matrix metalloproteinases or acting as signalling molecules, have always been produced with huge difficulties. We hereby propose a new method to overproduce human recombinant TIMP-1 by transient expression in HEK293E cells, followed by a one step chromatography purification, yielding in only 2 weeks, dozens of milligrams of pure, stable, glycosylated and active protein for in vitro and in vivo studies. This easy to set up, rapid, and efficient method could be applied for any naturally secreted mammalian protein. PMID- 16219295 TI - 5-Hydroxytryptamine augments migration of human aortic smooth muscle cells through activation of RhoA and ERK. AB - The purpose of this study was to elucidate the mechanism of 5-hydroxytryptamine (5-HT, serotonin) action on migration of vascular smooth muscle cells. Migration of cultured human aortic smooth muscle cells (HASMCs), evaluated using time-lapse microscopy, was significantly enhanced by 5-HT at concentrations of 1-100 nM. The enhancing effect of 5-HT on cell migration was markedly inhibited in the presence of ketanserin, a 5-HT2 receptor antagonist, but not by GR 55562, a 5-HT1 receptor antagonist. Activities of RhoA and ERK were increased by 5-HT, and the increase in cell migration by 5-HT was abolished in the presence of U0126, a MEK1/2 inhibitor, or Y-27632, a Rho-kinase inhibitor. Activation of ERK was strongly inhibited by Y-27632. 5-HT-induced formation of stress fiber and detachment of uropod (trailing edge) were abolished by Y-27632. Thus, 5-HT has a potent enhancing action on migration of HASMCs due to an increase in stress fiber formation by 5-HT2 receptor stimulation followed by activation of the Rho-kinase and ERK pathways. PMID- 16219296 TI - Autotaxin (lysoPLD/NPP2) protects fibroblasts from apoptosis through its enzymatic product, lysophosphatidic acid, utilizing albumin-bound substrate. AB - Autotaxin (ATX) was originally identified as a potent tumor cell motility stimulating factor that displays multiple enzymatic activities including ATPase, Type I nucleotide pyrophosphatase/phosphodiesterase, and lysophospholipase D, depending on its substrates. We demonstrate herein that ATX is a key regulator of extracellular lysophosphatidic acid (LPA) that can act as survival factor, in addition to its mitogenic activity in mouse fibroblasts. Introduction of atx gene into NIH3T3 cells resulted in resistance to conditional apoptosis induced by serum-deprivation, and exogenous ATX protein prevented cells from death by starvation. Flow cytometric analysis showed that co-treatment of ATX with lysophosphatidylcholine as substrate rescued NIH3T3 cells from cellular apoptosis, and this survival activity of ATX was also demonstrated by caspase-3 degradation and PARP cleavage resulting from the enzymatic activity of extracellular ATX. Furthermore, the effect of ATX in preventing apoptosis appears to be mediated through the G-protein-coupled receptor pathway followed by the activation of phosphoinositide 3-kinase and Akt pathway leading to enhanced cell survival. These findings provide novel insights into understanding the functions of ATX as a key regulator of bioactive phospholipids and suggest interventions to correct dysfunction in conditions of tumor cell growth and metastasis. PMID- 16219297 TI - Visualization of the trimeric P2X2 receptor with a crown-capped extracellular domain. AB - The P2X2 purinergic receptor permeates cationic ions in response to stimulation by ATP and mediates fast synaptic transmission. Here, we purified the P2X2 receptor using baculovirus-Sf9 cell expression system and observed its structure using electron microscopy. The FLAG-tagged P2X2 receptor, which has intact ion channel function, was purified to be a single peak by affinity purification and gel filtration chromatography. It was confirmed to be a trimer by introducing cross-linking. Negatively stained P2X2 protein images were homogeneous and picked up by automated pick-up programs, aligned, and classified using the modified growing neural gas network method. Similarly oriented projections were averaged to decrease the signal-to-noise ratio. These images demonstrate an inverted three sided pyramid with the dimensions of 215 A in height and 200 A in side length. It is composed of a high-density trunk and a stain-permeable swollen extracellular domain of a crown-shaped structure. The internal cavities and constituent segments were clearly demonstrated in both the raw images and the averaged images. The threefold symmetrical top view demonstrates the first visual evidence of the trimeric composition of the P2X receptor family. PMID- 16219298 TI - 3,3'-Diindolylmethane, a cruciferous vegetable derived synthetic anti proliferative compound in thyroid disease. AB - Considerable epidemiological evidence exists to link thyroid disease with differing patterns of dietary consumption, in particular, cruciferous vegetables. We have been studying the anti-thyroid cancer (TCa) activity of indole-3-carbinol (I3C) found in cruciferous vegetables and its acid catalyzed dimer, 3,3' diindolylmethane (DIM). There are no studies as yet to elucidate the effect of these compounds on the altered proliferative patterns in goiter or thyroid neoplasia. In this study, we tested the anti-proliferative effects of I3C and DIM on four different thyroid cancer cell lines representative of papillary (B-CPAP and 8505-C) and follicular carcinoma of the thyroid (CGTH-W-1 and ML-1), and primary human goiter cells. Cell survival and IC50 values for I3C and DIM were calculated by the XTT assay and cell cycle distribution analysis was done by flow cytometry. DIM was found to be a better anti-proliferative agent than I3C in both papillary and follicular TCa resulting in a greater cytotoxic effect at a concentration over three fold lower than predicted by the molar ratio of DIM and I3C. The anti-proliferative activity of DIM in follicular TCa was mediated by a G1 arrest followed by induction of apoptosis. DIM also inhibited the growth of primary goiter cells by 70% compared to untreated controls. Contrary to traditional belief that cruciferous vegetables are "goitrogenic", DIM has anti proliferative effects in glandular thyroid proliferative disease. Our preclinical studies provide a strong rationale for the clinical exploration of DIM as an adjuvant to surgery in thyroid proliferative disease. PMID- 16219299 TI - Inhibition of store-operated calcium entry-mediated superoxide generation by histamine trifluoromethyltoluide independent of histamine receptors. AB - Store-operated calcium entry (SOCE) plays an important role in shaping the Ca(2+) response of various tissues and cell types. In this report, we show that thapsigargin (TG)-induced SOCE was inhibited by the histamine receptor agonist, histamine-trifluoromethyltoluide (HTMT), in U937 and HL-60 human promyelocytes. Preincubation of HTMT resulted in a significant inhibition of subsequent TG induced Ca(2+) elevation without affecting Ca(2+) release from intracellular stores. HTMT also inhibited TG-induced Ca(2+) current and Ba(2+)/Mn(2+) influx in a concentration-dependent manner. In contrast with HTMT, other H1 histamine receptor agonists, histamine, 2-methylhistamine and 2-thiazolylethylamine, did not affect TG-induced SOCE. In addition, HTMT also attenuated TG-induced cytosolic superoxide generation. Taken together, our data clearly suggest that the anti-inflammatory effect of HTMT may occur through direct inhibition of SOCE. PMID- 16219300 TI - Pharmacological characterization of novel adenosine ligands in recombinant and native human A2B receptors. AB - The present study was designed to evaluate the effects of novel and recognised compounds at human recombinant A(2B) adenosine receptors expressed in Chinese hamster ovary (hA(2B)CHO), in human embryonic kidney 293 (hA(2B)HEK-293) and at endogenous A(2B) receptors in human mast cells (HMC-1). Saturation binding experiments performed using the new high affinity A(2B) adenosine radioligand [(3)H]-N-benzo[1,3]dioxol-5-yl-2-[5-(2,6-dioxo-1,3-dipropyl-2,3,6,7-tetra hydro 1H-purin-8-yl)-1-methyl-1H-pyrazol-3-yloxy]-acetamide ([(3)H]-MRE 2029F20) revealed a single class of binding sites in hA(2B)CHO, hA(2B)HEK-293 and HMC-1 cells with K(D) (nM) of 1.65+/-0.18, 2.83+/-0.34, 2.62+/-0.27 and B(max) (fmol/mg protein) of 36+/-4, 475+/-50 and 128+/-15, respectively. The pharmacological profile of new compounds, determined in inhibition binding experiments in hA(2B)HEK-293 cells using [(3)H]-MRE 2029F20, showed a rank order of potency typical of the A(2B) receptors with K(i) values in the range 3.2-28nM. In functional assays, recognised agonists and antagonists were studied by evaluating their capability to modulate the cAMP production in hA(2B)CHO and in HMC-1 cells. Novel compounds were able to decrease NECA-stimulated cAMP production in hA(2B)CHO and in HMC-1 cells showing a high potency. New compounds were also able to inhibit cAMP levels in the absence of NECA and in the presence of forskolin stimulation in hA(2B)CHO and in HMC-1 cells. In HEK-293 cells MRE 2029F20 reduced cAMP basal levels with an IC(50) value of 2.9+/-0.3nM. These results suggest that novel compounds are antagonists with an inverse agonist activity in recombinant and native human A(2B) receptors. PMID- 16219302 TI - Cardiac and peripheral actions of growth hormone and its releasing peptides: relevance for the treatment of cardiomyopathies. AB - Ischemic and nonischemic cardiomyopathies are associated with significant morbidity and mortality in industrialized countries. Cardiomyopathies of primary origin, and more specifically the dilated form of the disease, have been associated with a number of gene defects in cytoskeletal, membrane, and sarcomeric proteins. Cardiomyopathies of secondary origin such as ischemic cardiomyopathy remain the leading cause of left ventricular systolic dysfunction and heart failure. Among novel strategies to improve cardiac function in heart failure, treatment with growth hormone, insulin growth factor-1 (IGF-1), and natural and synthetic growth hormone-releasing peptides such as ghrelin and hexarelin have been explored. The present review focuses on the issues involved in the use of exogenous growth hormone and its releasing peptides in experimental animal models of chronic heart failure and in clinical studies on cardiomyopathic patients as potential releasing peptides for the treatment of chronic heart failure developing as a consequence of cardiomyopathy. PMID- 16219301 TI - IL-6 induces PI 3-kinase and nitric oxide-dependent protection and preserves mitochondrial function in cardiomyocytes. AB - OBJECTIVE: Interleukin-6 (IL-6) is a pro-inflammatory cytokine which is a prognostic marker associated with left ventricular contractile dysfunction and heart failure. On the other hand, IL-6 activates signalling pathways which mediate delayed ischemic preconditioning. We have therefore studied the cellular mechanisms of IL-6-induced cardioprotection. METHODS: Inducible nitric oxide synthase (iNOS) expression, cardiomyocyte calcium handling, mitochondrial energetics, and the activation of protective signalling pathways in response to IL-6 were studied in a model of simulated ischemia/reperfusion (sI/R) in isolated neonatal rat ventricular cardiomyocytes. RESULTS: Reperfusion after sI/R induced a rise in cytosolic [Ca2+], a loss of cell morphology and integrity, and a transient increase in mitochondrial potential (Deltapsi m), followed by mitochondrial swelling and collapse of Deltapsi m. Pre-treatment of cardiomyocytes with 10 ng/ml IL-6 for 6 h, 24 h prior to sI/R prevented the secondary rise in cytosolic [Ca2+] and induced expression of iNOS and NO dependent protection against sI/R injury. The protection against sI/R was concomitant with a NO-dependent reduction in the amplitude of cytosolic Ca2+ transients. IL-6 induced an increase in inner mitochondrial membrane polarisation and increased mitochondrial Ca2+ loading (rhod-2 fluorescence) at baseline, but prevented the reperfusion-induced changes in mitochondrial function. IL-6 pre treatment also resulted in activation of the phosphatidylinositol (PI) 3 kinase/Akt pathway, and both iNOS induction and IL-6-dependent protection were blocked by the PI 3-kinase inhibitor wortmannin. CONCLUSION: IL-6 induces a PI 3 kinase and NO-dependent protection of cardiomyocytes, which is associated with alterations in mitochondrial Ca2+ handling, inhibition of reperfusion-induced mitochondrial depolarisation, swelling and loss of structural integrity, and suppression of cytosolic Ca2+ transients. PMID- 16219303 TI - Panaxynol induces neurite outgrowth in PC12D cells via cAMP- and MAP kinase dependent mechanisms. AB - Panaxynol, a polyacetylene ((3R)-heptadeca-1,9-diene-4,6-diyn-3-ol; syn. falcarinol), was isolated from the lipophilic fractions of Panax notoginseng, a Chinese traditional medicinal plant. In the present study, we reported the neurotrophic effects of panaxynol on PC12D cells and mechanism involved in neurite outgrowth of the cells. Panaxynol could morphologically promote neurite outgrowth in PC12D cells, concentration-dependently reduce cell division and up regulate molecular marker (MAP1B) expression in PC12D cells. Panaxynol induces the elevation of intracellular cAMP in PC12D cells. The neurite outgrowth in PC12D cells induced by panaxynol could be inhibited by the protein kinase A inhibitor RpcAMPS and by MAP kinase kinase 1/2 inhibitor U0126. These observations reveal that panaxynol could induce the differentiation of PC12D cells in a process similar to but distinct from that of NGF and the panaxynol's effects were via cAMP- and MAP kinase-dependent mechanisms. PMID- 16219304 TI - 1,2-Dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) monolayers: influence of temperature, pH, ionic strength and binding of alkaline earth cations. AB - Ion binding and lipid ionization of the acidic phospholipid 1,2-dimyristoyl-sn glycero-3-phosphoglycerol (DMPG) in monolayers was studied by measuring the lateral pressure Pi as a function of the molecular area A at the air/water interface at different temperatures. The pH of the subphase (pH 2 and 7) and the ionic strength (NaCl) was varied. In addition, different divalent cations (1mM MgCl2, CaCl2 and SrCl2, pH 7) were added. DMPG is partly protonated on pure water at pH 7. An increase in the NaCl concentration in the subphase leads to film expansion. This effect is caused by an ionization of the headgroup of DMPG, i.e. a shift of the apparent pK. More condensed films are obtained on pure water at pH 2, due to the reduction of electrostatic repulsion by headgroup protonation and the possibility for the formation of a hydrogen bonding network. The divalent cations Mg2+, Ca2+ and Sr2+ interact differently with a DMPG monolayer in pure water at pH 7. In the presence of 1mM CaCl2 a condensation of the DMPG film is induced, whereas an expansion of the monolayer is observed in the presence of Mg2+ and Sr2+. Two counteracting effects are operative: (a) ionization of the headgroup due to electrostatic screening leads to film expansion and (b) binding of the divalent cations to the lipid headgroups leads to condensation. The latter effect is more pronounced in the case of Ca2+, whereas the binding of Mg2+ and Sr2+ to DMPG is weaker. Site-specific cation binding has to be assumed in addition to electrostatic effects. PMID- 16219305 TI - Hypomorphic Smn knockdown C2C12 myoblasts reveal intrinsic defects in myoblast fusion and myotube morphology. AB - Dosage of the survival motor neuron (SMN) protein has been directly correlated with the severity of disease in patients diagnosed with spinal muscular atrophy (SMA). It is also clear that SMA is a neurodegenerative disorder characterized by the degeneration of the alpha-motor neurons in the anterior horn of the spinal cord and atrophy of the associated skeletal muscle. What is more controversial is whether it is neuronal and/or muscle-cell-autonomous defects that are responsible for the disease per se. Although motor neuron degeneration is generally accepted as the primary event in SMA, intrinsic muscle defects in this disease have not been ruled out. To gain a better understanding of the influence of SMN protein dosage in muscle, we have generated a hypomorphic series of myoblast (C2C12) stable cell lines with variable Smn knockdown. We show that depletion of Smn in these cells resulted in a decrease in the number of nuclear 'gems' (gemini of coiled bodies), reduced proliferation with no increase in cell death, defects in myoblast fusion, and malformed myotubes. Importantly, the severity of these abnormalities is directly correlated with the decrease in Smn dosage. Taken together, our work supports the view that there is an intrinsic defect in skeletal muscle cells of SMA patients and that this defect contributes to the overall pathogenesis in this devastating disease. PMID- 16219306 TI - Transgenic mice expressing mutant (N279K) human tau show mutation dependent cognitive deficits without neurofibrillary tangle formation. AB - Mutations in the tau gene, which is located on chromosome 17, were found causative for autosomal dominantly inherited frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). To determine if cognitive deficits could be caused by tau mutations, two transgenic mouse lines were generated expressing a four-repeat isoform of human tau or its mutant, containing one of the FTDP-17 mutations (WILD mice and N279K mice). In open field test, N279K mice showed hyperactivity in locomotion and rearing. In prepulse inhibition test, N279K mice but not Wild mice showed significant deficits. Both transgenic mice, especially N279K mice, showed impairment in acquisition of spatial learning in Morris water maze. Although both N279K mice and Wild mice acquired passive avoidance as well as non-transgenic mice, N279K mice but not Wild mice showed severe deficits in acquisition of active avoidance. Histological analysis of the present mutant mice did not show any signs of neurofibrillary tangle formations in the brain, and cognitive dysfunction seemed to precede such neuropathological changes or occur independently from them. The behavioral phenotype of N279K mice mimics features of human FTDP-17 and provides a basic model for elucidating mechanisms underlying cognitive deficits in not only FTDP-17, but also diverse tauopathies. PMID- 16219307 TI - Intracellular processing of the Sendai virus C' protein leads to the generation of a Y protein module: structure-functional implications. AB - The Sendai virus "C-proteins" (C', C, Y1 and Y2) are a nested set of non structural proteins. The shorter Y proteins arise in vivo both by de novo translation initiation and by proteolytic processing of C'. In this paper, we demonstrate that C' but not C (differing only by 11 N-terminal amino acid) serves as an efficient substrate for intracellular processing. However, processing can be mimicked in vitro by the addition of endopeptidases. Under conditions of limited proteolysis we observed that in a fraction of the C' protein the Y region exists as a proteinase resistant core. This core was conserved in the C protein. We propose that C' functions as a Pro-protein delivering the Y module to a specific intracellular location. PMID- 16219308 TI - Cold-induced expression of the VEGF gene in brown adipose tissue is independent of thermogenic oxygen consumption. AB - To examine whether cold-induced vascular endothelial growth factor (VEGF) gene expression in brown adipose tissue involved generation of hypoxic oxygen levels by thermogenic processes, we cold-exposed wild-type mice, as well as uncoupling protein-1 (UCP1)-ablated mice in which no thermogenesis in brown adipocytes can be induced. Cold exposure stimulated VEGF expression in both wild-type and UCP1 ablated mice. Unexpectedly, the effect was 3-fold higher in UCP1-ablated animals, whereas cultured brown adipocytes from both genotypes responded identically to norepinephrine stimulation. These results demonstrate that generation of low oxygen levels does not contribute to cold-induced VEGF expression in brown adipose tissue, but the results are consistent with an adrenergic regulation of expression. PMID- 16219309 TI - Voltage-dependent synchronization of gating of syringomycin E ion channels. AB - Antifungal lipodepsipeptide syringomycin E (SRE) forms two major conductive states in lipid bilayers: "small" and "large". Large SRE channels are cluster of several small ones, demonstrating synchronous opening and closure. To get insight into the mechanism of such synchronization we investigated how transmembrane potential, membrane surface charge, and ionic strength affect the number of small SRE channels synchronously functioning in the cluster. Here, we report that the large SRE channels can be presented as 3-8 simultaneously gating small channels. The increase in the absolute value of the transmembrane potential (from 50 to 200 mV) decreases the number of synchronously gated channels in the clusters. Voltage dependence of channel synchronization was influenced by the ionic strength of the bathing solution, but not by membrane surface charge. We propose a mechanism for the voltage-dependent cluster behavior that involves a voltage-induced reorientation of lipid dipoles associated with the channel pores. PMID- 16219310 TI - Identification and analysis of Hic-5/ARA55 isoforms: Implications for integrin signaling and steroid hormone action. AB - Hic-5/ARA55 is a LIM-only member of the paxillin superfamily. Conflicting reports have suggested that Hic-5/ARA55 can both repress and enhance a number of biological processes, including myogenesis and tumorigenesis. With two Hic-5 isoforms documented, we hypothesized that multiple Hic-5 isoforms may exist that have both overlapping and isoform-specific functions. To test this hypothesis, we performed an extensive analysis of Hic-5 transcripts in both cell lines and mouse tissues and found 12 distinct isoforms that fall into two sub-families. These isoforms are derived from both alternative splicing and alternative transcriptional start sites (TSS). Hic-5 expression is regulated in a temporally and spatially controlled manner in vivo. The identification of numerous Hic-5 isoforms suggests that Hic-5 subsumes a number of distinct roles in cells and may explain the range of biological responses attributed to Hic-5. PMID- 16219311 TI - Validating the use of database potentials in protein structure determination by NMR. AB - The refinement of protein structures determined by nuclear magnetic resonance spectroscopy against database potentials of mean force allows for the exclusion of unfavourable conformations of the protein backbone during a structure calculation, resulting in protein structures with a marked improvement in Ramachandran statistics. In this communication, we use multiple sets of residual dipolar couplings as quality assessment criteria for several proteins and show that not only do the Ramachandran and structural quality statistics improve, but a significant improvement in the accuracy of structures is achieved upon refinement. PMID- 16219312 TI - Neither plasma coenzyme Q10 concentration, nor its decline during pravastatin therapy, is linked to recurrent cardiovascular disease events: a prospective case control study from the LIPID study. AB - Statins decrease LDL cholesterol and the risk of atherosclerotic cardiovascular disease (CVD). They also decrease coenzyme Q10 (CoQ10), an effect that may negate some of the statin benefit on CVD. We examined the relationship between plasma CoQ10 concentration and CVD in a prospective case-control study of the effect of pravastatin. Plasma samples from 250 LIPID trial patients who over 6 years suffered a recurrent CVD event (CVD death, nonfatal MI or stroke) and 250 matched controls who remained event-free for the same duration of follow-up were assayed for CoQ10 and lipids (cholesterol and cholesterylesters). Mean plasma CoQ10 concentrations were significantly lower in pravastatin-treated patients than in those assigned placebo (0.51 versus 0.60 micromol/L, P = 0.006), and there was a moderate correlation between CoQ10 and common cholesterylesters (Pearson correlation coefficients in patients randomised to placebo, range r = 0.42-0.63). Univariate conditional logistic regression did not suggest any relationship between plasma CoQ10 and the risk of future CVD events (odds ratio 1.18; 95% CI 0.74-1.87; P = 0.49). Instead, we observed a reduction in the rate of recurrent CVD events with increasing ratio of plasma cholesterylarachidonate to cholesteryllinoleate. This study confirms that pravastatin lowers plasma CoQ10 concentrations, but this does not appear to predict the risk of recurrent CVD events. PMID- 16219313 TI - Conjugated linoleic acids have no effect on TNF alpha-induced adhesion molecule expression, U937 monocyte adhesion, and chemokine release in human aortic endothelial cells. AB - Leukocyte recruitment and adhesion to the endothelium are critical steps in the early phase of atherosclerosis. Synthetic ligands of peroxisome proliferator activated receptors (PPARs) were shown to reduce cytokine-stimulated leukocyte endothelial cell interactions by inhibiting the NF-kappaB mediated inflammatory response. Conjugated linoleic acids (CLA), which are natural ligands of PPARs, were demonstrated to have anti-inflammatory and anti-atherogenic properties in vivo. With a view to elucidating the anti-atherogenic mechanisms of CLA, the present study aimed to explore the effects of cis-9, trans-11 CLA and trans-10, cis-12 CLA on cytokine-induced chemokine release, surface expression of adhesion molecules (ICAM-1, VCAM-1, and E-selectin) and U937 monocyte adhesion in human aortic endothelial cells (HAEC). Treatment of HAECs with 2 ng/mL of TNFalpha markedly increased expression of adhesion molecules, U937 monocyte adhesion, and release of the monocyte chemoattractant protein (MCP)-1. However, treatment of HAECs with either CLA isomer or linoleic acid did not modulate the cytokine induced expression of ICAM-1, VCAM-1, and E-selectin, U937 cell adhesion and MCP 1 release. In addition, both CLA isomers and linoleic acid slightly increased PPARgamma DNA-binding activity, but did not alter DNA-binding activity of NF kappaB. In conclusion, CLA isomers showed no effect on cytokine-induced monocyte endothelial cell interactions and on the molecular mechanisms regulating these processes in HAEC. This study suggests that anti-atherogenic effects of CLA observed in vivo are probably not mediated by reduced monocyte-endothelial cell interactions. PMID- 16219314 TI - Cross-correlation as a method for comparing dynamic electromyography signals during gait. AB - Current clinical interpretation of dynamic electromyography (EMG) data is usually based on qualitative assessments of muscle timing. Cross-correlation may provide a method for objectively comparing the timing and shape of EMG signals. This study used cross-correlation to compare EMG signals from different walking trials, different test sessions, and different individuals in able-bodied adults. Cross-correlation results (R-values) for different walking trials within a single test session were high, averaging > or = 0.90 for all muscles tested (R = 1.0 indicates exact agreement). Cross-correlation values were also high among trials from different test sessions conducted by the same and different examiners (average R > or = 0.78 for all muscles). R-values were much more variable when comparing different subjects (average 0.40-0.81, range 0.00-0.91). R-values were lower for the medial hamstrings and rectus femoris compared with the other muscles tested. These results suggest that cross-correlation may be useful for evaluating changes in an individual patient's muscle activation patterns, such as before and after surgery, but not for comparing EMG patterns among different individuals, such as between patients and normative data. This is especially true for biarticular muscles such as the hamstrings and rectus femoris, which may have variable activation patterns and/or increased sensitivity to electrode placement. Cross-correlation may also be useful for identifying appropriate muscles for transfer, identifying "outlier" trials within a test session, and selecting representative EMG curves for a given patient. The advantages of cross correlation are that it considers shape of the EMG signal in addition to timing and that the assessments it provides are objective, rather than subjective. PMID- 16219315 TI - Suitability of N,O-bis(trimethylsilyl)trifluoroacetamide as derivatization reagent for the determination of the estrogens estrone and 17 alpha ethinylestradiol by gas chromatography-mass spectrometry. PMID- 16219316 TI - Fixed-bed adsorption of chlorinated hydrocarbons from multicomponent aqueous solution onto activated carbon: equilibrium column model. AB - The results of studies on the adsorption dynamics of light chlorinated hydrocarbons, 1,2-dichloroethane, 1,1,2,2-tetrachloroethane, chloroform, carbon tetrachloride, 1,1-dichloroethene, perchloroethylene and 1,1,2-trichloroethene, from a seven-component solution on to activated carbon are presented. The experimental results were described using the equilibrium model. The application of this model allows to determine the location of the midpoint of the breakthrough profile. PMID- 16219317 TI - The characterization of prepared organomontmorillonite (DEDMAM) and sorption of phenoxyalkanoic acid herbicides from aqueous solution. AB - The montmorillonite has been subjected to modification through ion-exchange reaction by N,N'-didodecyl-N,N'-tetramethylethanediammoniumdiiodide (DEDMAI). The modified sample was studied by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and thermogravimetric analysis (TGA). The basal spacing of modified montmorillonite was determined as 28.72 A. The IR spectra of organomontmorillonite showed changes in C-H vibrations. The characterization of N,N'-didodecyl-N,N'-tetramethylethanediammonium montmorillonite (DEDMAM) and the adsorption of 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4-dichlorophenoxybutyric acid (2,4-DB), 4-chloro-2-methylphenoxyacetic acid (MCPA), and 2,4,5 trichlorophenoxyacetic acid (2,4,5-T) on organomontmorillonite was studied as a function of the solution concentration. The adsorption energy (E) and adsorption capacity (qm) for phenoxyalkanoic acid herbicides adsorbing on organomontmorillonite (DEDMAM) were estimated using the Dubinin-Radushkevic (D-R) equation. These isotherms were modeled according to Freundlich and Dubinin Radushkevic adsorption isotherms. The sorption of the herbicides on DEDMAM increased in order of MCPA<2,4-D<2,4-DB<2,4,5-T. PMID- 16219318 TI - Experimental and modeling study of Newtonian and non-Newtonian fluid flow in pore network micromodels. AB - The in situ rheology of polymeric solutions has been studied experimentally in etched silicon micromodels which are idealizations of porous media. The rectangular channels in these etched networks have dimensions typical of pore sizes in sandstone rocks. Pressure drop/flow rate relations have been measured for water and non-Newtonian hydrolyzed-polyacrylamide (HPAM) solutions in both individual straight rectangular capillaries and in networks of such capillaries. Results from these experiments have been analyzed using pore-scale network modeling incorporating the non-Newtonian fluid mechanics of a Carreau fluid. Quantitative agreement is seen between the experiments and the network calculations in the Newtonian and shear-thinning flow regions demonstrating that the 'shift factor,'alpha, can be calculated a priori. Shear-thickening behavior was observed at higher flow rates in the micromodel experiments as a result of elastic effects becoming important and this remains to be incorporated in the network model. PMID- 16219320 TI - Crystal structures of an intein from the split dnaE gene of Synechocystis sp. PCC6803 reveal the catalytic model without the penultimate histidine and the mechanism of zinc ion inhibition of protein splicing. AB - The first naturally occurring split intein was found in the dnaE gene of Synechocystis sp. PCC6803 and belongs to a subclass of inteins without a penultimate histidine residue. We describe two high-resolution crystal structures, one derived from an excised Ssp DnaE intein and the second from a splicing-deficient precursor protein. The X-ray structures indicate that His147 in the conserved block F activates the side-chain N(delta) atom of the intein C terminal Asn159, leading to a nucleophilic attack on the peptide bond carbonyl carbon atom at the C-terminal splice site. In this process, Arg73 appears to stabilize the transition state by interacting with the carbonyl oxygen atom of the scissile bond. Arg73 also seems to substitute for the conserved penultimate histidine residue in the formation of an oxyanion hole, as previously identified in other inteins. The finding that the precursor structure contains a zinc ion chelating the highly conserved Cys160 and Asp140 reveals the structural basis of Zn2+-mediated inhibition of protein splicing. Furthermore, it is of interest to observe that the carbonyl carbon atom of Asn159 and N(eta) of Arg73 are 2.6 angstroms apart in the free intein structure and 10.6 angstroms apart in the precursor structure. The orientation change of the aromatic ring of Tyr-1 following the initial acyl shift may be a key switching event contributing to the alignment of Arg73 and the C-terminal scissile bond, and may explain the sequential reaction property of the Ssp DnaE intein. PMID- 16219319 TI - IgG subclass-independent improvement of antibody-dependent cellular cytotoxicity by fucose removal from Asn297-linked oligosaccharides. AB - Fucose depletion from oligosaccharides of human IgG1-type antibodies results in a great enhancement of antibody-dependent cellular cytotoxicity (ADCC). The aim of this study was to clarify the effect of fucose removal on effector functions of all human IgG subclasses. A panel of anti-CD20 chimeric antibodies having a matched set of human heavy chain subclasses with different fucose contents in their oligosaccharides was constructed using wild-type and fucosyltransferase knockout Chinese hamster ovary cells as host cells. As found previously for IgG1, fucose-negative variant of IgG2, IgG3, and IgG4 exhibited enhanced ADCC and FcgammaRIIIa binding compared with their highly fucosylated counterparts. In contrast, fucose removal did not affect complement-dependent cytotoxicity (CDC) of any IgGs. Consequently, fucose removal from IgG2 and IgG4 resulted in a unique effector function profile; they had potent ADCC and no CDC. In conclusion fucose depletion can provide a panel of IgGs with enhanced ADCC without an impact on other inherent properties specific for each IgG subclass, such as CDC. PMID- 16219321 TI - Structural characterization of the catalytic subunit of a novel RNA splicing endonuclease. AB - The RNA splicing endonuclease is responsible for recognition and excision of nuclear tRNA and all archaeal introns. Despite the conserved RNA cleavage chemistry and a similar enzyme assembly, currently known splicing endonuclease families have limited RNA specificity. Different from previously characterized splicing endonucleases in Archaea, the splicing endonuclease from archaeum Sulfolobus solfataricus was found to contain two different subunits and accept a broader range of substrates. Here, we report a crystal structure of the catalytic subunit of the S.solfataricus endonuclease at 3.1 angstroms resolution. The structure, together with analytical ultracentrifugation analysis, identifies the catalytic subunit as an inactive but stable homodimer, thus suggesting the possibility of two modes of functional assembly for the active enzyme. PMID- 16219322 TI - Crystal structures of the main peptidase from the SARS coronavirus inhibited by a substrate-like aza-peptide epoxide. AB - The main peptidase (M(pro)) from the coronavirus (CoV) causing severe acute respiratory syndrome (SARS) is one of the most attractive molecular targets for the development of anti-SARS agents. We report the irreversible inhibition of SARS-CoV M(pro) by an aza-peptide epoxide (APE; k(inact)/K(i) = 1900(+/-400) M( 1) s(-1)). The crystal structures of the M(pro):APE complex in the space groups C2 and P2(1)2(1)2(1) revealed the formation of a covalent bond between the catalytic Cys145 S(gamma) atom of the peptidase and the epoxide C3 atom of the inhibitor, substantiating the mode of action of this class of cysteine-peptidase inhibitors. The aza-peptide component of APE binds in the substrate-binding regions of M(pro) in a substrate-like manner, with excellent structural and chemical complementarity. In addition, the crystal structure of unbound M(pro) in the space group C2 revealed that the "N-fingers" (N-terminal residues 1 to 7) of both protomers of M(pro) are well defined and the substrate-binding regions of both protomers are in the catalytically competent conformation at the crystallization pH of 6.5, contrary to the previously determined crystal structures of unbound M(pro) in the space group P2(1). PMID- 16219323 TI - Symmetric connectivity of secondary structure elements enhances the diversity of folding pathways. AB - The influence of native connectivity of secondary structure elements (SSE) on folding is studied using coarse-grained models of proteins with mixed alpha and beta structure and the analysis of the structural database of wild-type proteins. We found that the distribution of SSE along a sequence determines the diversity of folding pathways. If alpha and beta SSE are localized in different parts of a sequence, the diversity of folding pathways is restricted. An even (symmetric) distribution of alpha and beta SSE with respect to sequence midpoint favors multiple folding routes. Simulations are supplemented by the database analysis of the distribution of SSE in wild-type protein sequences. On an average, two-thirds of wild-type proteins with mixed alpha and beta structure have symmetric distribution of alpha and beta SSE. The propensity for symmetric distribution of SSE is especially evident for large proteins with the number of SSE > or = 10. We suggest that symmetric SSE distribution in protein sequences may arise due to nearly random allocation of alpha and beta structure along wild-type sequences. The tendency of long sequences to misfold is perhaps compensated by the enhanced pathway diversity. In addition, folding pathways are shown to progress via hierarchic assembly of SSE in accordance with their proximity along a sequence. We demonstrate that under mild denaturation conditions folding and unfolding pathways are similar. However, the reversibility of folding/unfolding pathways is shown to depend on the distribution of SSE. If alpha and beta SSE are localized in different parts of a sequence, folding and unfolding pathways are likely to coincide. PMID- 16219324 TI - Visualization of the phosphorylated active site loop of the cytoplasmic B domain of the mannitol transporter II(Mannitol) of the Escherichia coli phosphotransferase system by NMR spectroscopy and residual dipolar couplings. AB - The solution structure of a stably phosphorylated form of the cytoplasmic B domain of the mannitol-specific transporter (IIB(Mtl)) of the Escherichia coli phosphotransferase system, containing a mutation of the active site Cys384 to Ser, has been solved by NMR. The strategy employed relies principally on backbone residual dipolar couplings recorded in three different alignment media, supplemented by nuclear Overhauser enhancement data and torsion angle restraints related specifically to the active site loop (residues 383-393). As judged from the dipolar coupling data, the remainder of the structure is unchanged upon phosphorylation within the errors of the coordinates of the previously determined solution structure of unphosphorylated wild-type IIB(Mtl). Thus, only the active site loop was refined. Phosphorylation results in a backbone atomic rms shift of approximately 0.7 angstroms in the active site loop. The resulting conformation is less than 0.5 angstroms away from the equivalent P-loop in both the low and high molecular mass eukaryotic tyrosine phosphatases. 3J(NP) coupling constant measurements using quantitative J-correlation spectroscopy provide a direct demonstration of a hydrogen bond between the phosphoryl group and the backbone amide of Ser391 at position i + 7 from phospho-Ser384, with an approximately linear P-O-H(N) bond angle. The structure also reveals additional hydrogen bonding interactions involving the backbone amides of residues at positions i + 4 and i + 5, and the hydroxyl groups of two serine residues at positions i + 6 and i + 7 that stabilize the phosphoryl group. PMID- 16219325 TI - Distinct spatial and temporal distribution of ZAP70 and Lck following stimulation of interferon and T-cell receptors. AB - T-cell receptor (TCR) stimulation results in the recruitment and activation of the proteins ZAP70 and Lck. These two proteins have been implicated in signalling derived from interferon receptors, although their precise role in this independent pathway has not been determined fully. These observations raise a fundamental question of how a given protein in a cell can be involved in more than one signalling pathway, yet each pathway is able to produce a highly specific downstream response to its own stimulant. To maintain exclusivity of response, each pathway must isolate its component molecules chemically, spatially or dynamically from other prevailing pathways. To address this question, the proteins ZAP70 and Lck were investigated following stimulation of the interferon alpha receptor and the TCR in T cells by two different extracellular stimulants: interferon-alpha and the anti-CD3 antibody, OKT3, respectively. We first demonstrate that ZAP70 plays a pivotal role in interferon-stimulated MAPK activation, and that the tyrosine residue at position 319 of ZAP70 is important for interferon-stimulated ERK activation. Translocation of both ZAP70 and Lck to the nucleus following interferon receptor stimulation is demonstrated for the first time. Fluorescence resonance energy transfer microscopy revealed a high degree of spatial localization of the ZAP70/Lck complex within the cell following IFNalpha stimulation, in contrast to a diffuse presence following the application of OKT3. The difference in the spatio-temporal localization of these proteins following stimulation may eliminate signal crosstalk, and could explain the differentiation of the specific downstream responses of these pathways. PMID- 16219326 TI - N-terminally truncated variant of the mouse GAIP/RGS19 lacks selectivity of full length GAIP/RGS19 protein in regulating ORL1 receptor signaling. AB - The regulators of G protein signaling (RGS) are a family of proteins with conserved RGS domains and play essential roles in regulating G protein-mediated signal transduction and physiological events. GAIP/RGS19 (G alpha interacting protein, also classified as RGS19), a member of the RGS family, has been shown to negatively regulate the signaling of many G protein-coupled receptors, including the opioid receptors. Two GAIP/RGS19 mRNA variants, resulted from an alternative splicing of exon 2 of the GAIP/RGS19 gene, were identified in multiple mouse tissues. One of the transcripts consists of a complete set of exons and encodes a full-length GAIP/RGS19 protein, and the other does not have exon 2 and therefore encodes an N-terminal 22 residue truncated short GAIP/RGS19 protein. When co expressed with either the opioid-receptor-like (ORL1) receptor or one of the mu, delta, and kappa opioid receptors, by transfecting dual-expression plasmids into COS-7 cells, the full-length GAIP/RGS19 was more effective than the N-terminally truncated variant and was more selective in regulating the ORL1 receptor signaling than in regulating the mu, delta, and kappa opioid receptors, as measured by the effectiveness to increase the agonist-stimulated GTPase activity and to reverse the agonist-induced inhibition of cyclic AMP accumulation. In the same assays, the N-terminally truncated GAIP/RGS19 did not distinguish ORL1 from the mu, delta, and kappa opioid receptors. In contrast, co-expression of RGS4 with either ORL1 or opioid receptors showed the selectivity of RGS4 for regulating opioid receptors was mu > kappa > delta > ORL1, an order completely different from that of GAIP/RGS19. The results suggest that GAIP/RGS19 prefers regulating ORL1 receptor signaling over other opioid receptors, and that the N terminal domain of GAIP/RGS19 plays a crucial role in its receptor preference. PMID- 16219328 TI - Spreading disease with transport-related infection. AB - Transportation among regions is found as one of the main factors which affect the outbreak of diseases. It will change the disease dynamics and break infection out even if infectious diseases will go to extinction in each city without transport related infection. In this paper, a mathematical model is proposed to demonstrate the dynamics of such disease propagation between two regions (or cities) due to the population dispersal and infection on transports. Further, our analysis shows that transport-related infection intensifies the disease spread if infectious diseases break out to cause an endemic situation in each region, in the sense that both the absolute and relative size of patients increase. This suggests that it is very essential to strengthen restrictions of passengers once we know infectious diseases appeared. PMID- 16219327 TI - Plasma lipid peroxidation in sporadic Parkinson's disease. Role of the L-dopa. AB - Oxidative stress plays an important role in the pathogenesis of neurodegenerative diseases, such as Parkinson's disease (PD). There are several methods to measure oxidative stress, being lipid peroxidation (LPO) one of the most frequently used. Endogenous plasma LPO was determined by a spectrofluorimetric method in fifty two patients with sporadic PD and in forty controls. To know the maximum capacity of lipids to peroxidate, LPO was also measured after co-incubation with Fe2+/H2O2 (exogenous LPO). All PD patients were taken L-dopa and the effect of this treatment on LPO levels was additionally studied. Urine catecholamines and their main metabolites were also analyzed, and their possible correlation to LPO statistically studied. Endogenous plasma LPO levels were 33% higher in PD group than in control group (P<0.001). Exogenous plasma or oxidizability was also higher in PD patients compared to controls (20%, P<0.05). The intake of L-dopa was negatively dose-related to endogenous and exogenous plasma LPO. In conclusion, plasma of PD patients has elevated levels of LPO and also is more prone to peroxidation than that in the control group. The results also suggest an antioxidant effect of L-dopa. PMID- 16219329 TI - Diffusion versus network models as descriptions for the spread of prion diseases in the brain. AB - In this paper we will discuss different modeling approaches for the spread of prion diseases in the brain. Firstly, we will compare reaction-diffusion models with models of epidemic diseases on networks. The solutions of the resulting reaction-diffusion equations exhibit traveling wave behavior on a one-dimensional domain, and the wave speed can be estimated. The models can be tested for diffusion-driven (Turing) instability, which could present a possible mechanism for the formation of plaques. We also show that the reaction-diffusion systems are capable of reproducing experimental data on prion spread in the mouse visual system. Secondly, we study classical epidemic models on networks, and use these models to study the influence of the network topology on the disease progression. PMID- 16219330 TI - Schistosoma mansoni in pregnancy and associations with anaemia in northwest Tanzania. AB - Schistosomiasis among pregnant women has been inadequately investigated. In order to determine the importance of Schistosoma mansoni in this subgroup, we conducted a cross-sectional survey of 972 women in Tanzania and investigated the prevalence of Schistosoma mansoni, hookworm and malaria and their associations with anaemia. Overall, 63.5% of women were infected with S. mansoni, with prevalence highest among younger women and decreasing with increasing age. The prevalence of hookworm was 56.3%, and 16.4% of women had malaria parasitaemia. Overall, 66.4% of women were anaemic. Increased risk of anaemia was associated with heavy infection with S. mansoni but not hookworm or Plasmodium falciparum parasitaemia. PMID- 16219331 TI - Mutations other than 103N in human immunodeficiency virus type 1 reverse transcriptase (RT) emerge from K103R polymorphism under non-nucleoside RT inhibitor pressure. AB - K103N mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) confers high-level resistance against non-nucleoside RT inhibitors (NNRTIs) and it easily occurs partly because it arises by a single nucleotide substitution from wild-type K103. There are polymorphisms at codon 103 of HIV-1 RT. We found K103R polymorphic mutation in 3.3% of treatment-naive HIV-1 infected patients. R103N does not seem to occur as easily as K103N because R103N requires two nucleotide substitutions. To induce NNRTI resistance-associated mutations, HIV-1K103R was propagated in the presence of increasing concentrations of efavirez (EFV) or nevirapine (NVP). V179D emerged in all three EFV cultures and in two of four NVP cultures. R103G emerged by a single nucleotide substitution in one of three EFV cultures. R103N did not emerge in any of 7 NNRTI cultures. Analysis of recombinant HIV-1s showed that HIV-1K103R/V179D was significantly resistant and HIV-1K103G was moderately resistant against EFV and NVP. PMID- 16219332 TI - Escherichia coli O157:H7 in drinking water from private water supplies in the Netherlands. AB - The microbiological quality of drinking water from 144 private water supplies in the Netherlands was tested and additionally the occurrence of Escherichia coli O157 was examined. Faecal indicators were enumerated by using standard membrane filtration methods. The presence of E. coli O157 was determined using a specific enrichment method. Eleven percent of the samples contained faecal indicators whereas E. coli O157:H7 was isolated from 2.7% of the samples that otherwise met the drinking water standards. The E. coli O157 positive water supplies were located on camp-sites in agricultural areas with large grazer densities. Pulsed field gel electrophoresis (PFGE) analysis suggested that cattle might have been the cause of contamination. Our results indicate that compliance with microbiological quality standards obtained in routine monitoring does not always guarantee the absence of pathogens. The presence of pathogens such as E. coli O157 may suggest possible health consequences; however, a risk assessment process should be performed as the monitoring of both faecal indicator parameters and pathogens do not predict the effect of microbial contamination of drinking water on a population. PMID- 16219333 TI - Influence of geometrical and operational parameters on the axial dispersion in an aerated channel reactor. AB - Residence time distribution experiments have been performed on an activated sludge 3000 m3 channel reactor aerated by gas diffusion (for different liquid flowrates under constant aeration rate and constant water depth) and on a bench scale channel reactor aerated from the bottom (for different liquid and gas flowrates and water depths) in order to characterize their hydrodynamics. Both units can be modeled as plug flow reactors with axial dispersion. A general correlation has been obtained to predict the axial dispersion coefficient as a function of the gas and liquid velocities and the geometrical parameters of the full-scale and bench-scale reactors. Finally, to facilitate the simulation of biological reactions in transient state, an equivalent model based on tanks-in series with variable back-mixing flowrate is proposed. PMID- 16219334 TI - Laccase-catalyzed removal of bisphenol-A from water: protective effect of PEG on enzyme activity. AB - The feasibility of using the enzyme laccase to treat synthetic wastewater containing bisphenol-A (BPA) was examined. Optimization of pH, laccase concentration, polyethylene glycol (PEG) as an additive for >95% conversion and precipitation of BPA over 3 h of reaction period was determined through colorimetric assay and HPLC. PEG reduced enzyme inactivation, allowing a 5.2-fold reduction in the amount of laccase required for >95% removal of BPA in the range of 0.1-1 mM over 3 h. The fate of PEG after the reaction was also monitored. Linear relationships were found between the concentration of BPA (0.1-1 mM) and the optimum concentrations of laccase and PEG. Little PEG remained in the solution when up to 75 mg/L of PEG was used to treat 0.5 mM BPA. Beyond this level, PEG concentration increased linearly in the supernatant. It is inferred that an interaction between PEG and the polymeric products resulted in the protection of laccase. PMID- 16219335 TI - Monitoring of effluent DOM biodegradation using fluorescence, UV and DOC measurements. AB - The potential of effluent DOM to undergo microbial degradation was assessed in batch experiments. Effluent samples from Haifa wastewater treatment plant and Qishon reservoir (Greater Haifa wastewater reclamation complex, Israel) were incubated either with effluent or soil microorganisms for a period of 2-4 months and were characterized by dissolved organic carbon contents (DOC), UV(254) absorbance and by fluorescence excitation-emission matrices. Three main fluorescence peaks were identified that can be attributed to humic/fulvic components and "protein-like" structures. During biodegradation, specific fluorescences (F/DOC) of the three peaks were increased at various extents, suggesting selective degradation of non-fluorescing constituents. In some cases increase in the effluent fluorescence (F) was observed thus proposing (i) the formation of new fluorescing material associated with DOM biodegradation and/or (ii) degradation of certain organic components capable of quenching DOM fluorescence. Based on the ratio between fluorescence intensity and UV(254), different biodegradation dynamics for fluorescent DOM constituents as compared with other UV-absorbing molecules was delineated. Overall, about 50% of the total DOM was found to be readily degradable such that residual resistant DOC levels were between 8 and 10 mg l(-1). Enhanced levels of residual DOM in effluent irrigated soils may contribute to the DOM pool capable of carrying pollutants to groundwater. PMID- 16219336 TI - Effect of iron on lipid peroxidation, and enzymatic and non-enzymatic antioxidants and bacoside-A content in medicinal plant Bacopa monnieri L. AB - The effect of Fe was investigated in medicinally important plant, Bacopa monnieri L. and the response on malondialdehyde (MDA) content, superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) was found different in roots and leaves of the metal treated plants. Iron induced stress was observed as indicated by high level of lipid peroxidation, being more steep increase in leaves than roots. In roots, SOD activity was found to increase in metal treated plants except 80 and 160 microM at 72 h, whereas, it decreased in leaves except 10 and 40 microM after 48 h as compared to their respective controls. Among H2O2 eliminating enzymes, POD activity increased in roots, however, it decreased in leaves except at 10 and 40 microM Fe after 48 h as compared to control. At 24 and 48 h, APX activity and ascorbic acid content followed the similar trend and were found to increase in both parts of the metal treated plants as compared to their respective controls. The level of cysteine content in the roots increased at initial period of exposure; however, no marked change in its content was noticed in leaves. In both roots and leaves, non-protein thiol content was found to increase except at higher metal concentrations at 72 h. The data of proline content have shown significant (p<0.01) increase at 40 microM onwards in both part of the plants after 48 and 72 h. Correlation coefficient was evaluated between metal accumulations with various parameters and also between different antioxidant parameters with MDA. Since the level of bacoside-A (active constituent) content in metal treated plants increases, therefore, it is advisable to assess the biological activity of the plants before using for medicinal purposes, particularly in developing countries. PMID- 16219337 TI - The cross-sectional geometry of the hand and foot bones of the hominoidea and its relationship to locomotor behavior. AB - Cheiridia are valuable indicators of positional behavior, as they directly contact the substrate, but systematic comparison of the structural properties of both metacarpals and metatarsals has never been carried out. Differences in locomotor behavior among the great apes (knuckle-walking vs. quadrumanous climbing) can produce biomechanical differences that may be elucidated by the parallel study of cross-sectional characteristics of metacarpals and metatarsals. The aim of this work is to study the cross-sectional geometric properties of these bones and their correlation with locomotor behavior in large-bodied hominoids. The comparisons between bending moments of metacarpals and metatarsals of the same ray furnished interesting results. Metacarpals III and especially IV of the knuckle-walking African apes were relatively stronger than those of humans and orangutans, and metatarsal V of humans was relatively stronger than those of the great apes. Interestingly, the relative robusticity of the metacarpal IV of the quadrumanous orangutan was between that of the African apes and that of humans. The main conclusions of the study are: 1) cross-sectional dimensions of metacarpals and metatarsals are influenced by locomotor modes in great apes and humans; 2) interlimb comparisons of cross-sectional properties of metacarpals and metatarsals are good indicators of locomotor modes in great apes and humans; and 3) the results of this study are in accord with those of previous analyses of plantar pressure and morphofunctional traits of the same bones, and with behavioral studies. These results provide a data base from which it will be possible to compare the morphology of the fossils in order to gain insight into the locomotor repertoires of extinct taxa. PMID- 16219338 TI - Influence of intensive fishing on the partitioning of mercury and methylmercury in three lakes of Northern Quebec. AB - It has been demonstrated that intensive fishing, i.e., removing more than 25% of the fish biomass, can reduce mercury levels in predator fish in a lake. We test here the hypothesis that, by removing an important part of the fish biomass from a lake, a significant amount of methylmercury can be eliminated, therefore reducing the mercury available to the remaining biota, at least in the short term. A mass burden approach is used to evaluate the partitioning of total mercury and methylmercury in natural lake ecosystems. Three small natural lakes from the James Bay territory, in northern Quebec, Canada, were selected for intensive fishing. Mercury (Hg) and methylmercury (MeHg) concentrations were evaluated for sediments, water column (dissolved fraction and suspended particulate matter), plankton, aquatic invertebrates, and fish. Biomasses were determined for fish, plankton, and aquatic invertebrates. Two case scenarios are presented using different mercury contributions from the sediment component (1 cm depth, and no sediment). Our results for the scenario including the sediment contribution show that lake sediments represent over 98% of the total mercury while the biotic components represent less than 0.1% of the same burden. For methylmercury, fish account for up to 5% of the burden, while sediments make up 84.6% to 93.1%. If we put aside the sediment contribution, the methylmercury in fish partitioning can represent up to 48%. As for invertebrates, they can account for up to 48% of the total MeHg burden. We do not observe any change in the partitionings or the quantities of Hg and MeHg before and after fishing in either of the two case scenarios even when we do not take into account dynamics of the ecosystems. This will be all the more the case when the dynamics of the system are included in the analyses. Therefore, biological parameters such as growth rates or fish diet must be considered. PMID- 16219339 TI - Contaminants in Canadian arctic biota and implications for human health: conclusions and knowledge gaps. AB - This paper summarizes the major findings of the special issue entitled "Contaminants in Canadian Arctic Biota and Implications for Human Health." The individual papers and reviews in this special issue present a large amount of new information on contaminants in biota primarily from the Canadian arctic as well as from Alaska, Greenland and the European Arctic. Temporal and spatial trends are examined and potential biological effects on wildlife are assessed. The special issue also presents new and updated data on human exposure to and possible health effects of current levels of environmental contaminants in the Canadian Arctic. As part of the assessment of the human health implications, the unique structures and processes that have developed in the Canadian Arctic under the Northern Contaminants Program (NCP) of Indian and Northern Affairs Canada to build partnerships and manage and communicate the benefits and risks associated with contaminant exposure are discussed. Application of this information in international forums to reduce anthropogenic emissions of contaminants to the environment is also discussed. PMID- 16219340 TI - Estimation of mercury emission from different sources to atmosphere in Chongqing, China. AB - This investigation presents a first assessment of the contribution to the regional mercury budget from anthropogenic and natural sources in Chongqing, an important industrial region in southwest China. The emissions of mercury to atmosphere from anthropogenic sources in the region were estimated through indirect approaches, i.e. using commonly acceptable emission factors method, which based on annual process throughputs or consumption for these sources. The natural mercury emissions were estimated from selected natural sources by the dynamic flux chamber technique. The results indicated that the anthropogenic mercury emissions totaled approximately 8.85 tons (t), more than 50% of this total originated in coal combustion and 23.7% of this total emission in the industrial process (include cement production, metal smelting and chemical industry). The natural emissions represented approximately 17% of total emissions (1.78 t yr(-1)). The total mercury emission to atmosphere in Chongqing in 2001 was 10.63 t. PMID- 16219341 TI - Analysis of the QUESTOR water quality model using a Fourier amplitude sensitivity test (FAST) for two UK rivers. AB - This paper presents the sensitivity analysis of a well-known in-stream water quality model, QUESTOR (QUality Evaluation and Simulation TOol for River systems) as applied to two rivers of contrasting land-use in the northeast of England: the 'rural' Ouse and the 'urban' Aire. The analysis employed a version of the Fourier Amplitude Sensitivity Test (FAST) that quantifies the contribution of changes in individual parameters and combination of parameters to the variance of the model output (here the Nash-Sutcliffe) in an efficient way. The quantification of the sensitivity of the model output to the parameters led to the identification of the most influential parameters. Differences between the Aire and the Ouse were found, reflecting their different water quality regime. Results highlighted the importance of interactions between two, or more, parameters on the model output. It led to question the one-at-a-time calibration method currently applied with QUESTOR and underlined the importance of including interactions between parameters in sensitivity analyses. Comparison of the relative influence of parameters versus input data showed contrasting results. In the urban system, the inputs from discharges (sewage treatment works and industrial effluents) were highly influential on model outputs and generally more important than the model parameters. For the rural river, the tributary discharges were most influential, but only at a similar or a lower level than the model parameters. PMID- 16219342 TI - Influence of fluoxetine and paroxetine in behavioral sensitization induced by ethanol in mice. AB - The serotonergic system is involved in depression, anxiety and alcoholism. The rewarding properties of ethanol, mainly its anxiolytic and stimulant effects, as well as the development of dependence on ethanol have been related to the serotonergic system. Consequently, the use of selective serotonergic reuptake inhibitors (SSRI) has been proposed in the treatment of alcoholism. In this study we investigated whether acute administration of the SSRIs fluoxetine or paroxetine is able to (i) reverse the behavioral effects induced by chronic ethanol consumption, and conversely, (ii) to determine whether acute ethanol is able to substitute for the chronically induced behavioral effects of fluoxetine or paroxetine. Four groups of male Swiss mice (n=60/group) received daily i.p. saline, ethanol (2 g/kg), fluoxetine (10 mg/kg) or paroxetine (5 mg/kg) for 27 days. On the 28th day, each group was challenged with saline, ethanol, fluoxetine or paroxetine. The 14 groups (SS, SE, SP, SF, EE, ES, EP, EF, PP, PE, PS, FF, FE, and FS) were then tested in open field, activity cage and plus-maze. EP and EF groups were able to reverse the behavioral sensitization to the psychomotor stimulant effects of chronic ethanol administration. In contrast, a sensitized stimulatory effect was observed in chronically fluoxetine- or paroxetine treated mice challenged with ethanol (PE and FE). An anxiolytic effect was observed whether ethanol was substituted for SSRI or, conversely, SSRI was substituted for ethanol. SSRIs facilitated ethanol-induced locomotor sensitization, although SSRIs by themselves are unable to produce the locomotor stimulation similar to that induced by ethanol. Finally, SSRIs are unable to interfere in the ethanol anxiolytic effect. PMID- 16219343 TI - The use of a deslorelin implant (GnRH agonist) during the late embryonic period to reduce pregnancy loss. AB - Embryonic and fetal mortality reduce reproductive performance of lactating dairy cows. The objectives of this study were to reduce pregnancy loss by administering a deslorelin implant (GnRH agonist) during the late embryonic period, to reduce follicular growth, induce accessory corpora lutea, and increase plasma progesterone concentrations. Lactating dairy cows received an implant containing 2.1 mg of deslorelin (Deslorelin group; n = 89) or no treatment (Control group; n = 92) on Day 27 of pregnancy. Pregnancy, ovarian structures and plasma progesterone concentrations were determined on Days 27 and 45, and pregnancy was re-confirmed on Day 90. On Day 45, mean +/- S.E.M. numbers of class 2 (6-9 mm; 0.72+/-0.19) and class 3 (> or = 10 mm; 0.86 +/- 0.12) follicles for cows in the Deslorelin group were lower (P < 0.01) than the numbers of class 2 (1.90 +/- 0.18) and class 3 (1.92 +/- 0.12) follicles for cows in the Control group. On Day 45, the number of accessory corpora lutea for cows in the Deslorelin group (1.80 +/- 0.07) were greater (P < 0.01) than for cows in the Control group (1.31 +/- 0.07). On Day 45, plasma progesterone concentration was increased (P < 0.01) for cows in the Deslorelin group (8.03 +/- 0.33 ng/mL) compared to cows in the Control group (6.40 +/- 0.31 ng/mL). Pregnancy losses did not differ between Days 27 and 45 and Days 45 and 90 for cows in the Control (15.2 and 11.0%, respectively) and Deslorelin groups (20.2 and 10.5%, respectively). However, in the Deslorelin group, pregnancy loss between Days 45 and 90 was lower (P < 0.05) for cows that formed an accessory CL (0%) compared to cows that did not form an accessory CL (16.1%). PMID- 16219344 TI - Binding of the bioactive compound 5,7,4'-trihydroxy-6,3',5'-trimethoxyflavone to human serum albumin. AB - 5,7,4'-trihydroxy-6,3',5'-trimethoxyflavone is one of the bioactive components isolated from Artemisia plants possessing antitumor therapeutic activities. In this paper, its binding properties and binding sites located on human serum albumin (HSA) have been studied using UV absorption spectroscopy, fluorescence spectroscopy and Fourier transform infrared (FT-IR) spectra. The results of fluorescence titration revealed that 5,7,4'-trihydroxy-6,3',5'-trimethoxyflavone could strongly quench the intrinsic fluorescence of HSA by static quenching and there was only one class of binding sites on HSA for this drug. The binding constants at four different temperatures (289, 298, 310, and 318 K) were 1.93, 1.56, 1.22, and 0.93x10(5) L mol-1, respectively. The FT-IR spectra evidence showed that the protein secondary structure changed with reduction of alpha helices about 27.6% at the drug to protein molar ratio of 3. The thermodynamic functions standard enthalpy change (DeltaH0) and standard entropy change (DeltaS0) for the reaction were calculated to be -18.70 kJ mol-1 and 36.62 J mol 1 K-1 according to the van't Hoff equation. These results and the molecular modeling study suggested that hydrophobic interaction was the predominant intermolecular force stabilizing the complex, and 5,7,4'-trihydroxy-6,3',5' trimethoxyflavone could bind to the site I of HSA (the Warfarin Binding site). PMID- 16219345 TI - Preparation, physical-chemical characterisation and cytocompatibility of calcium carbonate cements. AB - The feasibility of calcium carbonate cements involving the recrystallisation of metastable calcium carbonate varieties has been demonstrated. Calcium carbonate cement compositions presented in this paper can be prepared straightforwardly by simply mixing water (liquid phase) with two calcium carbonate phases (solid phase) which can be easily obtained by precipitation. An original cement composition was obtained by mixing amorphous calcium carbonate and vaterite with an aqueous medium. The cement set and hardened within 2h at 37 degrees C in an atmosphere saturated with water and the final composition of the cement consisted mostly of aragonite. The hardened cement was microporous and showed poor mechanical properties. Cytotoxicity tests revealed excellent cytocompatibility of calcium carbonate cement compositions. Calcium carbonates with a higher solubility than the apatite formed for most of the marketed calcium phosphate cements might be of interest to increase biomedical cement resorption rates and to favour its replacement by bone tissue. PMID- 16219346 TI - A method for solvent-free fabrication of porous polymer using solid-state foaming and ultrasound for tissue engineering applications. AB - Most of the existing fabrication techniques for tissue engineering scaffolds require the use of organic solvents that may never be fully removed even after long leaching hours. The residues of these organic solvents reduce the ability of biological cells to form new tissue. This paper presents an approach toward solvent-free fabrication of tissue engineering scaffolds. Interconnected porous structures were created using solid-state foaming and ultrasound. The material used in this study was polylactic acid (PLA) and the blowing agent was CO(2). In order to determine suitable process conditions, saturation and foaming studies were first conducted. Selected foam samples were then processed using pulsed ultrasound. The microstructures before and after the ultrasound processing were compared. It was shown that the inter-pore connectivity of the solid-state foams was substantially enhanced. The combined solid-state foaming and ultrasound processing provide a way to fabricate porous polymer for potential tissue engineering applications. PMID- 16219347 TI - Attenuated alloreactivity of dendritic cells engineered with surface-modified microspheres carrying a plasmid encoding interleukin-10. AB - In the present study, we investigated MS(O10H6) as a carrier system to introduce a plasmid encoding murine interleukin-10 (pIL-10) to modulate alloreactivity of dendritic cells (DC). Results indicate that MS(O10H6) formed stable and protective nano-sized particles with pIL-10. Gene-modified DC elicited weak proliferation of allogeneic CD4 and CD8 T cells in vitro. Using cell-embedded Matrigel as a surrogate graft, we also showed that DC transfected with MS(O10H6) complexed with pIL-10 suppressed host cell infiltration in vivo. These data demonstrate that the self-assembled system of MS(O10H6) is an effectual delivery vehicle for plasmid-based modulation of DC-dependent allogeneic T cell responses. PMID- 16219348 TI - Phthalic acid diamides activate ryanodine-sensitive Ca2+ release channels in insects. AB - Flubendiamide represents a novel chemical family of substituted phthalic acid diamides with potent insecticidal activity. So far, the molecular target and the mechanism of action were not known. Here we present for the first time evidence that phthalic acid diamides activate ryanodine-sensitive intracellular calcium release channels (ryanodine receptors, RyR) in insects. With Ca(2+) measurements, we showed that flubendiamide and related compounds induced ryanodine-sensitive cytosolic calcium transients that were independent of the extracellular calcium concentration in isolated neurons from the pest insect Heliothis virescens as well as in transfected CHO cells expressing the ryanodine receptor from Drosophila melanogaster. Binding studies on microsomal membranes from Heliothis flight muscles revealed that flubendiamide and related compounds interacted with a site distinct from the ryanodine binding site and disrupted the calcium regulation of ryanodine binding by an allosteric mechanism. This novel insecticide mode of action seems to be restricted to specific RyR subtypes because the phthalic acid diamides reported here had almost no effect on mammalian type 1 ryanodine receptors. PMID- 16219350 TI - Combination of erythropoietin and thalidomide for the treatment of anemia in patients with myelodysplastic syndromes. AB - We investigated the therapeutic activity of recombinant erythropoietin (r-EPO) in association with thalidomide in 30 patients with myelodysplastic syndromes (MDS), previously treated with r-EPO (n.15, group A) or thalidomide (n.15, group B) as single agents, respectively, without any significant benefit on their anemia. Four patients of group A and three of group B (23.3%) achieved an erythroid response, according to International Working Group (IWG) criteria. After 12 weeks, responders of group A continued with thalidomide alone, those of group B with r-EPO alone. All responses were maintained, thus suggesting they were likely due to the second drug adjuncted (thalidomide for group A and r-EPO for group B), rather than to a combined effect. Our results do not support the hypothesis of a synergistic activity for the association of r-EPO and thalidomide on anemia of MDS. It seems, instead, that two populations of patients can be identified, according to their sensitivity to r-EPO or, alternatively, to thalidomide. PMID- 16219349 TI - Antigen-induced Ca2+ mobilization in RBL-2H3 cells: role of I(1,4,5)P3 and S1P and necessity of I(1,4,5)P3 production. AB - Inositol 1,4,5-trisphosphate (IP3) has long been recognized as a second messenger for intracellular Ca2+ mobilization. Recently, sphingosine 1-phosphate (S1P) has been shown to be involved in Ca2+ release from the endoplasmic reticulum (ER). Here, we investigated the role of S1P and IP3 in antigen (Ag)-induced intracellular Ca2+ mobilization in RBL-2H3 mast cells. Antigen-induced intracellular Ca2+ mobilization was only partially inhibited by the sphingosine kinase inhibitor dl-threo-dihydrosphingosine (DHS) or the IP3 receptor inhibitor 2-aminoethoxydiphenyl borate (2-APB), whereas preincubation with both inhibitors led to complete inhibition. In contrast, stimulation of A3 adenosine receptors with N5-ethylcarboxamidoadenosine (NECA) caused intracellular Ca2+ mobilization that was completely abolished by 2-APB but not by DHS, suggesting that NECA required only the IP3 pathway, while antigen used both the IP3 and S1P pathways. Interestingly, however, inhibition of IP3 production with the phospholipase C inhibitor U73122 completely abolished Ca2+ release from the ER induced by either stimulant. This suggested that S1P alone, without concomitant production of IP3, would not cause intracellular Ca2+ mobilization. This was further demonstrated in some clones of RBL-2H3 cells excessively overexpressing a beta isoform of Class II phosphatidylinositol 3-kinase (PI3KC2beta). In such clones including clone 5A4C, PI3KC2beta was overexpressed throughout the cell, although endogenous PI3KC2beta was normally expressed only in the ER. Overexpression of PI3KC2beta in the cytosol and the PM led to depletion of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), resulting in a marked reduction in IP3 production. This could explain the abolishment of intracellular Ca2+ mobilization in clone 5A4C. Supporting this hypothesis, the Ca2+ mobilization was reconstituted by the addition of exogenous PI(4,5)P2 in these cells. Our results suggest that both IP3 and S1P contribute to FcvarepsilonRI-induced Ca2+ release from the ER and production of IP3 is necessary for S1P to cause Ca2+ mobilization from the ER. PMID- 16219351 TI - Histologic and molecular characterizations of megakaryocytic leukemia in mice. AB - Six cases of megakaryocytic leukemia (MKL) were identified and analyzed for morphology and molecular features. MKL were composed of megakaryocyte lineage cells ranging from immature to quite mature cells. VWF, GATA1 and RUNX1 were strongly expressed in megakaryocytes in both normal spleen and MKL as analyzed by immunohistochemistry (IHC). Altered expression of Meis1, Pbx1 and Psen2 and Lef1 in MKL detected with oligonucleotide microarrays was confirmed by qPCR and IHC. This is the first report of spontaneous MKL in mice, defining VWF as a biomarker for diagnosis and suggesting possible involvement of a series of genes in disease pathogenesis. PMID- 16219352 TI - Combination of tetrandrine as a potential-reversing agent with daunorubicin, etoposide and cytarabine for the treatment of refractory and relapsed acute myelogenous leukemia. AB - The potential mechanism of the chemotherapy resistance in acute myeloid leukemia (AML) is the multidrug resistance (MDR-1) gene product P-glycoprotein (P-gp), which is often overexpressed in myeloblasts from acute myeloid leukemia. In a multicenter clinical trial, 38 patients with poor risk forms of AML were treated with tetrandrine (TET), a potent inhibitor of the MDR-1 efflux pump, combined with daunorubicin (DNR), etoposide and cytarabine (TET-DEC). Overall, post chemotherapy marrow hypoplasia was achieved in 36 patients. Sixteen patients (42%) achieved complete remission or restored chronic phase, 9 achieved partial remission (PR) and 13 failed therapy. Toxicities included infection, myelosuppression, stomatitis, mucositis, cerebellar toxicity and reversible cardiotoxicity. There was no significant difference in response for P-gp-positive and -negative patients. P-gp function was assessed in 26 patients by flow cytometric analysis, TET-contained plasma-augmented DNR accumulation relative to pretreatment plasma in K562/A02 cells by a median value of 88+/-101% (range, 11 501%). However, there was no difference in DNR uptake between responding and non responding patients. Our data showed that TET-DEC was relatively well tolerated in these patients with poor risk AML, and had encouraging antileukemic effects. PMID- 16219353 TI - The 5q- syndrome and autoimmune phenomena: report of three cases. AB - Myelodysplastic syndrome is a clonal hematopoietic stem cell disorder characterized by ineffective hematopoiesis, peripheral cytopenias and an additional risk to evolve to acute leukemia in up to 30% of the cases. Autoimmune manifestations as vasculitis, pyoderma gangrenosum, hemolytic anemia, immune thrombocytopenia, rheumatoid arthritis as well as positive anti-nuclear factor and rheumatoid factor have been reported in 13-30% of MDS patients. The aim of this report is to present three patients with 5q- syndrome who presented different autoimmune serological and clinical phenomena and review the literature. Patient 1 showed a focal and segmental glomerulosclerosis (FSGE) in the course of a MDS. Renal involvement in MDS as autoimmune phenomenon is rare and few reports have documented different forms of glomerular diseases in adults with MDS. Patients 2 and 3 showed a rheumatoid factor of 1/140 and the direct Coomb's test positive (3+), respectively, but without evidence of clinical autoimmune manifestation. In conclusion, patients with the 5q- syndrome experience a relative benign disease course extending over several years. We believe that careful follow-up of patients with autoimmune manifestations as here reported is important to detect any unexpected outcome. PMID- 16219354 TI - The cytidine deaminase AID exhibits similar functional properties in yeast and mammals. AB - A recent work published in Molecular Immunology examined the editing activity of activation-induced deaminase (AID) in yeast (Krause, K., Marcu, K.B., Greeve, J., 2006. The cytidine deaminases AID and APOBEC-1 exhibit distinct functional properties in a novel yeast selectable system. Mol. Immunol.). It was proposed that expression of AID in yeast is not sufficient for the generation of point mutations in a highly transcribed gene due to the lack of cofactors for AID induced somatic hypermutation, which are unique to B cells. It was suggested that, on its own, AID does not have an intrinsic specificity for its target sequences. However, it has been shown previously that expression of the human AID gene in yeast was moderately mutagenic in a wild-type strain and highly mutagenic in an ung1 uracil-DNA glycosylase-deficient strain (Mayorov, V.I., Rogozin, I.B., Adkison, L.R., Frahm, C.R., Kunkel T.A., Pavlov Y. I., 2005. Expression of human AID in yeast induces mutations in context similar to the context of somatic hypermutation at G-C pairs in immunoglobulin genes. BMC Immunol. 6, 10; Poltoratsky, V.P., Wilson, S.H., Kunkel, T.A., Pavlov, Y.I., 2004. Recombinogenic phenotype of human activation-induced cytosine deaminase. J. Immunol. 172, 4308 4313). The vast majority of mutations were at G-C pairs. Mutations showed a clear DNA sequence context specificity which resembled the specificity of somatic hypermutation at G-C pairs in immunoglobulin genes and AID mutation specificity in vitro. The inability to detect mutator effects of AID by Krause et al. is likely to be caused by the use of the wild-type yeast strain and a small sample of clones examined for the presence of mutations. In addition, we show that non uniformity of the mutation hotspot distribution is a factor potentially decreasing the chances of detecting mutations. PMID- 16219355 TI - Agreement among 3 optical imaging methods for the assessment of optic disc topography. AB - PURPOSE: To assess the agreement of disc topography measurements between the Heidelberg Retina Tomograph (HRT II), Retinal Thickness Analyzer (RTA), and Optical Coherence Tomograph (StratusOCT). DESIGN: Observational cross-sectional study. PARTICIPANTS: Forty-two randomly chosen eyes of 42 subjects. METHODS: Each subject underwent HRT II, RTA, and StratusOCT examination. Two experienced examiners drew the contour lines for the HRT II and RTA. Bland and Altman plots were used to evaluate agreement for each topographic parameter among the instruments. The Spearman coefficient of rank correlation was evaluated for each topographic parameter. MAIN OUTCOME MEASURES: Agreement in the measurement of optic disc topography among 3 imaging instruments, as evaluated by regression based 95% limits of agreement. RESULTS: For optic disc area, the agreement between HRT II-RTA and StratusOCT-RTA revealed the existence of proportional bias, indicated by significant slopes of the regression lines (P = 0.01 and P = 0.02, respectively). The 95% limits of agreement between instruments varied with the actual optic disc size measurement. Heidelberg Retina Tomograph disc area measurements tended to be consistently lower than StratusOCT disc area measurements (fixed bias). The Spearman correlation coefficient between the instruments ranged from r = 0.35 (rim area, HRT II-StratusOCT) to r = 0.91 (cup area, HRT II-RTA). CONCLUSIONS: Moderate to high correlation was found in measurements of optic disc topography among different instruments. However, the analysis of agreement indicated important discrepancies among instruments. Therefore, these instruments should not be used interchangeably to obtain measurements of the optic disc for glaucoma diagnosis. PMID- 16219356 TI - Fate of manganese associated with the inhalation of welding fumes: potential neurological effects. AB - Welding fumes are a complex mixture composed of different metals. Most welding fumes contain a small percentage of manganese. There is an emerging concern among occupational health officials about the potential neurological effects associated with the exposure to manganese in welding fumes. Little is known about the fate of manganese that is complexed with other metals in the welding particles after inhalation. Depending on the welding process and the composition of the welding electrode, manganese may be present in different oxidation states and have different solubility properties. These differences may affect the biological responses to manganese after the inhalation of welding fumes. Manganese intoxication and the associated neurological symptoms have been reported in individual cases of welders who have been exposed to high concentrations of manganese-containing welding fumes due to work in poorly ventilated areas. However, the question remains as to whether welders who are exposed to low levels of welding fumes over long periods of time are at risk for the development of neurological diseases. For the most part, questions remain unanswered. There is still paucity of adequate scientific reports on welders who suffered significant neurotoxicity, hence there is a need for well-designed epidemiology studies that combine complete information on the occupational exposure of welders with both behavioral and biochemical endpoints of neurotoxicity. PMID- 16219357 TI - Studies on the interaction mechanism between hexakis(imidazole) manganese(II) terephthalate and DNA and preparation of DNA electrochemical sensor. AB - The interaction between hexakis(imidazole) manganese(II) terephthalate ([Mn(Im)(6)](teph).4H(2)O) and salmon sperm DNA in 0.2M pH 2.30 Britton-Robinson buffer solution was studied by fluorescence spectroscopy and cyclic voltammetry. Increasing fluorescence was observed for [Mn(Im)(6)](2+) with DNA addition, while quenching fluorescence phenomenon appeared for EB-DNA system when [Mn(Im)(6)](2+) was added. There were a couple quasi-reversible redox peaks of [Mn(Im)(6)](2+) from the cyclic voltammogram on the glassy carbon electrode. The peak current of [Mn(Im)(6)](2+) decreased with positive shift of the formal potential in the presence of DNA compared with that in the absence of DNA. All the experimental results indicate that [Mn(Im)(6)](2+) can bind to DNA mainly by intercalative binding mode. The binding ratio of the DNA-[Mn(Im)(6)](2+) association complex is calculated to be 1:1 and the binding constant is 4.44x10(3) M(-1). By using [Mn(Im)(6)](teph).4H(2)O as the electrochemical hybridization indicator, the DNA electrochemical sensor was prepared by covalent interaction and the selectivity of ssDNA modified electrode were described. The results demonstrate the use of electrochemical DNA biosensor in the determination of complementary ssDNA. PMID- 16219358 TI - Synthesis, characterization and biological activity of oxovanadium (IV) complexes with cyclic polyalcohols. AB - Oxovanadium (IV) complexes of the cyclic polyols conduritol C (cond) and myo inositol (inos) of stoichiometry Na(2)[VO(cond)(2)].2H(2)O and Na(2)[VO(inos)(2)].H(2)O were obtained in aqueous alkaline solutions. They were characterized by infrared and UV-Vis spectroscopies, thermoanalytical (thermogravimetric and differential thermal analysis) data and magnetic susceptibility measurements. The biological activities of the complexes on the proliferation, differentiation and glucose consumption were tested on osteoblast like cells in culture. Conduritol C and myo-inositol did not produce any effect on these parameters. Normal and tumoral cell proliferation was inhibited about (ca.40-60%) by the two oxovanadium (IV) complexes in concentrations as low as 100microM. The complexes were also inhibitory on cell differentiation (ca. 70 80%) while they stimulate glucose consumption. Comparisons of these effects with those of the oxovanadium (IV) cation, under the same experimental conditions, were also performed. PMID- 16219359 TI - Interactions of vanadium(V)-citrate complexes with the sarcoplasmic reticulum calcium pump. AB - Among the biotargets interacting with vanadium is the calcium pump from the sarcoplasmic reticulum (SR). To this end, initial research efforts were launched with two vanadium(V)-citrate complexes, namely (NH(4))(6)[V(2)O(4)(C(6)H(4)O(7))(2)].6H(2)O and (NH(4))(6)[V(2)O(2)(O(2))(2)(C(6)H(4)O(7))(2)].4H(2)O, potentially capable of interacting with the SR calcium pump by combining kinetic studies with (51)V NMR spectroscopy. Upon dissolution in the reaction medium (concentration range: 4 0.5mM), both vanadium(V):citrate (VC) and peroxovanadium(V):citrate (PVC) complexes are partially converted into vanadate oligomers. A 1mM solution of the PVC complex, containing 184microM of the PVC complex, 94microM oxoperoxovanadium(V) (PV) species, 222microM monomeric (V1), 43microM dimeric (V2) and 53microM tetrameric (V4) species, inhibits Ca(2+) accumulation by 75 %, whereas a solution of the VC complex of the same vanadium concentration, containing 98microM of the VC complex, 263microM monomeric (V1), 64microM dimeric (V2) and 92microM tetrameric (V4) species inhibits the calcium pump activity by 33 %. In contrast, a 1 mM metavanadate solution, containing 460microM monomeric (V1), 90.2microM dimeric (V2) and 80microM tetrameric (V4) species, has no effect on Ca(2+) accumulation. The NMR signals from the VC complex (-548.0ppm), PVC complex (-551.5ppm) and PV (-611.1ppm) are broadened upon SR vesicle addition (2.5mg/ml total protein). The relative order for the half width line broadening of the NMR signals, which reflect the interaction with the protein, was found to be V4>PVC>VC>PV>V2=V1=1, with no effect observed for the V1 and V2 signals. Putting it all together the effects of two vanadium(V)-citrate complexes on the modulation of calcium accumulation and ATP hydrolysis by the SR calcium pump reflected the observed variable reactivity into the nature of key species forming upon dissolution of the title complexes in the reaction media. PMID- 16219360 TI - Cerebral diffusional changes in the early phase of anthrax: is cutaneous anthrax only limited to skin? AB - OBJECTIVES: Characteristics of cerebral diffusion in the acute period of the anthrax infection were investigated to understand the pathophysiology of the disease. METHODS: Six cutaneous anthrax patients (mean age: 33.3, SD: 18.1) and six healthy control subjects (mean age: 33.7, SS: 19.6) were examined at the acute phase of the infection with diffusion weighted imaging on 1.5 T scanner. ADC values were measured from five different cerebral locations. T-tests, logistic regression and ROC curves were used. RESULTS: Anthrax patients were significantly different than controls regarding cortical ADC values (p<0.05). Logistic regression model accurately classified five out of the six anthrax cases (83.3%). A cut-off value of 574 mm2/s x 10(-3) was found by using ROC curve coordinates. A sensitivity of 100% and a specificity of 67% were attained by means of this value. CONCLUSIONS: This study shows the existence of cerebral parenchymal changes at microstructural level in cutaneous anthrax without neurological findings. These changes are possibly related to the components of the toxin. Our results support the general but unproven opinion that anthrax treatment does not change the existence and the effects of the toxin. Pathophysiological mechanisms towards classification should therefore be reviewed. PMID- 16219361 TI - Endothelin receptor antagonists. AB - Endothelin receptor antagonists (ERAs) have been developed to block the effects of endothelin-1 (ET-1) in a variety of cardiovascular conditions. ET-1 is a powerful vasoconstrictor with mitogenic or co-mitogenic properties, which acts through the stimulation of 2 subtypes of receptors [endothelin receptor subtype A (ETA) and endothelin receptor subtype B (ETB) receptors]. Endogenous ET-1 is involved in a variety of conditions including systemic and pulmonary hypertension (PH), congestive heart failure (CHF), vascular remodeling (restenosis, atherosclerosis), renal failure, cancer, and cerebrovascular disease. The first dual ETA/ETB receptor blocker, bosentan, has already been approved by the Food and Drug Administration for the treatment of pulmonary arterial hypertension (PAH). Trials of endothelin receptor antagonists in heart failure have been completed with mixed results so far. Studies are ongoing on the effects of selective ETA antagonists or dual ETA/ETB antagonists in lung fibrosis, cancer, and subarachnoid hemorrhage. While non-peptidic ET-1 receptor antagonists suitable for oral intake with excellent bioavailability have become available, proven efficacy is limited to pulmonary hypertension, but it is possible that these agents might find a place in the treatment of several cardiovascular and non-cardiovascular diseases in the coming future. PMID- 16219362 TI - Gender differences in adolescent depression: do symptoms differ for boys and girls? AB - BACKGROUND: Limited prior research suggests that depressed women are more likely to experience certain symptoms of depression than are depressed men. The purpose of this study was to examine whether such gender differences in depressive symptoms are present during adolescence. METHODS: The Childhood Version of the Schedule for Affective Disorders and Schizophrenia and the Beck Depression Inventory were administered to adolescents presenting for evaluation at an outpatient clinic (n=383; ages 11.9 to 20.0). RESULTS: Depressed girls and boys had similar symptom prevalence and severity ratings for most depressive symptoms. However, depressed girls had more guilt, body image dissatisfaction, self-blame, self-disappointment, feelings of failure, concentration problems, difficulty working, sadness/depressed mood, sleep problems, fatigue, and health worries than depressed boys on some comparisons. In contrast, depressed boys had higher clinician ratings of anhedonia, depressed morning mood, and morning fatigue. LIMITATIONS: Longitudinal research is needed to test whether such relatively gender-specific symptoms play different roles in the onset, maintenance, or remittance of depression for boys and girls. CONCLUSIONS: These findings indicate that, in general, the experience of depression is highly similar for adolescent girls and boys. However, some gender differences previously found among depressed adults appear to be present by adolescence, possibly suggesting somewhat distinct etiologies for depression among males and females. PMID- 16219363 TI - Immune responses after local administration of IgY loaded-PLGA microspheres in gut-associated lymphoid tissue in pigs. AB - Oral vaccination of large animals using PLGA MS (poly(D,L-lactide-co glycolide)microspheres) appeared to be more challenging than immunization of mice. The purpose of this study was to deliver to GALT an immunogenic model protein (IgY), free or encapsulated by spray-drying in PLGA MS, and to evaluate systemic immune response in SPF Large White pigs. Pigs were surgically processed for local administration of IgY in three sets of experiments. In two sets of experiments, administration was locally performed in temporary ligatured intestinal segments, in jejunal Peyer's patches and in mesenteric lymph nodes. In the third experiment, pigs received IgY via an intestinal cannula. Total IgY specific antibodies were detected in the sera of pigs after a single local immunization, but not in the sera of cannulated pigs. The study of IgG1 and IgG2 isotypes indicated that PLGA MS are able to elicit a combined serum IgG2/G1 response with a predominance of IgG1 response when locally administered. PLGA MS can be a potential oral delivery system for antigen but our results underlined the difficulty to immunize large animals like pigs. Transposition of data between small and large animals appears to be complex and suggests that physiological features need to be considered to increase intestinal availability of oral encapsulated vaccines. PMID- 16219364 TI - Membrane IgM influences membrane IgD mediated antigen internalization in the B cell line Bcl1. AB - Signalling through the B cell antigen receptor (BCR) is required for peripheral B lymphocyte maturation, maintenance, activation and silencing. In mature B cells, the antigen receptor normally consists of two isotypes: membrane IgM and IgD (mIgM, mIgD). Although the signals initiated from both isotypes differ in kinetics and intensity, in vivo, the BCR of either isotype seems to be able to compensate for the loss of the other, reflected by the mild phenotypes of mice deficient for mIgM or mIgD. Thus, it is still unclear why mature B cells need expression of mIgD in addition to mIgM. In the present paper, we used the B cell line Bcl1 and investigated the isotype-specific antigen internalization in dependence of co-stimulation of the reciprocal isotype and analysed whether the signal initiated from mIgM is modulated through signalling from mIgD and vice versa. We clearly showed that cross-linkage of mIgM decreases the rate of mIgD mediated antigen internalization and interpret this influence as a unilateral mIgM mediated control on signals initiated at mIgD. PMID- 16219365 TI - Re: Cytogenetic biomonitoring in children with chronic tonsillitis. PMID- 16219367 TI - An immunoassay for the pathological form of the prion protein based on denaturation and time resolved fluorometry. AB - Concern about the possible secondary spread of variant Creutzfeldt-Jakob disease (vCJD) through blood transfusion and blood products has increased the need for a sensitive and rapid test for the identification of PrP(Sc) in specimens collected non-invasively from living persons. Furthermore, an accurate estimate of the prevalence of pre-clinical vCJD in the British population would be possible if there were such a test that could be applied to specimens available readily (e.g. blood and urine). As a first step towards that goal, we have developed a simple and sensitive test for the detection of PrP(Sc) in peripheral tissues and brain of vCJD patients, based on the differential extraction of PrP(Sc) with guanidine hydrochloride. The prion protein (PrP) isoforms are extracted sequentially from homogenized tissue by applying two different concentrations of this chaotropic agent. Each extraction yields a fraction of the PrP isoforms with different solubilities in guanidine hydrochloride. Quantitation of the two fractions (relatively insoluble or relatively soluble) using time resolved fluorescence (DELFIA) as a reporter system allows differentiation between PrP(Sc) infected and non-infected tissues. The assay has a detection limit of 10 pg PrP, is robust and could be automated. PMID- 16219368 TI - Evaluation of dendrimer SPL7013, a lead microbicide candidate against herpes simplex viruses. AB - Dendrimers are a novel class of polyanionic macromolecules with broad-spectrum antiviral activities and minimal toxicities. A new generation of amide dendrimer, SPL7013, was evaluated as a lead microbicide candidate against herpes simplex viruses (HSV). The plaque reduction assays showed that the 50% effective concentrations (EC(50)) determined by pre-treatment of cells were 2.0 microg/ml for HSV-1 and 0.5 microg/ml for HSV-2. Inhibitory effects were also observed on HSV-infected cells with EC(50)s of 6.1 microg/ml for HSV-1 and 3.8 microg/ml for HSV-2. These are the mean values from the test results of six batches of SPL7013. SPL7013 was also shown to be equally potent against HSV drug-resistant strains. SPL7013 completely inhibited viral adsorption to Vero cells at concentrations of higher than 3 microg/ml. Analyzed by a LightCycler assay after treatment of HSV infected cells for 17 h, SPL7013 showed strong inhibition of HSV DNA synthesis with EC(50)s of approximately 6.2 and 2.0 microg/ml for HSV-1 and HSV-2, respectively. SPL7013 retained its anti-HSV activity even after treatment at acidic pHs 3.0 and 4.0 for 2 h. The presence of 10% human serum proteins did not affect the anti-HSV activity of SPL7013. SPL7013 was not toxic to Vero cells up to the highest concentration tested (10,000 microg/ml). Effects on cell proliferation were tested on two epithelial cell lines in both stationary and dividing phases. The 50% cytotoxic concentrations (CC(50)) in all cases were greater than 10,000 microg/ml. Our data indicate that SPL7013 is a promising candidate for development as a vaginal microbicide and a therapeutic agent. PMID- 16219369 TI - Arousal from hibernation alters contextual learning and memory. AB - Hibernation is a unique and highly regulated physiological state characterized by profound, albeit periodically reversible, depression in body temperature, metabolism, and consciousness. Hippocampal synapses undergo pronounced remodeling in concert with torpor and arousal. During hibernation, the number of postsynaptic densities, apical dendritic branches, and spine densities decreases substantially in the hippocampus. Upon arousal these parameters increase beyond pre-hibernation levels and peak within 2-3h. By 24h after arousal, dendritic parameters remain elevated but have started to subside, consistent with pruning and differentiation. The present study examined the functional consequences of these natural changes in synaptic structure. Wild-caught Arctic ground squirrels (AGS) were trained in a hippocampal-dependent contextual fear conditioning task at 3h, 24h, or 4 weeks after arousal (warm-adapted euthermic control group). All groups acquired the fear conditioned response similarly on the training day. During a subsequent retention test session, AGS in the 24h group exhibited enhanced expression of contextual fear compared to the other two groups. These data suggest that the morphological and biochemical changes occurring at 24h after arousal from hibernation affect hippocampal-dependent learning and memory. The natural change in synaptic structure during hibernation may provide a unique opportunity to assess the neural substrates underlying cognitive enhancement. PMID- 16219370 TI - Effects of 17beta-estradiol, 4-nonylphenol and PCB 126 on the estrogenic activity and phase 1 and 2 biotransformation enzymes in male sea bass (Dicentrarchus labrax). AB - The endocrine system of wildlife is exposed to a wide variety of natural and man made chemicals which may lead to damage to the reproductive system and other adverse effects, including alteration of drug-metabolizing enzymes. In the present study, the effects of in vivo exposure to a natural (17beta-estradiol: E2) or a xenoestrogen (4-nonylphenol: NP) estrogen or an anti-estrogen (3,3',4,4',5-pentachlorobiphenyl: PCB 126) upon vitellogenin (VTG) synthesis and hepatic phase 1 and 2 enzymes have been investigated in adult male sea bass. By means of ELISA analysis with the use of polyclonal antibodies prepared against VTG purified from E2-treated sea bass, we assessed the time course and sensitivity of VTG induction in the plasma of sea bass treated with E2 at 0.1, 0.5, 2.5 and 5.0 mg/kg doses or NP at 5.0 or 50 mg/kg doses, respectively. Sea bass sensitivity to this induction was found to be similar to that of other fish species, but with a delay in maximal response. E2 treatment also caused a selective time- and dose-dependent inhibition of hepatic CYP1A-linked EROD and phase 2 glutathione S-transferase (GST) activities, without affecting the activity of CYP3A-linked 6beta-testosterone hydroxylase, (omega)- and (omega-1) lauric acid hydroxylases or phase 2 DT-diaphorase. A similar selective inhibition on CYP1A was also observed in fish treated with 50 mg/kg NP. The results regarding CYP1A and CYP3A were also confirmed by Western blot analysis. When the sea bass were treated with either 10 or 100 microg/kg PCB 126, an AhR ligand not yet tested in vivo in fish to assess its anti-estrogenicity, a modest and selective induction of EROD and DT-diaphorase activities was observed. Interestingly, both these activities were recovered to their control levels in sea bass co-treated with 0.5 mg/kg E2 and 10 or 100 microg/kg PCB 126, probably through a cross-talk mechanism between the estrogen receptor and AhR or other transcription factors that regulate the expression of these enzymes. Furthermore, it was demonstrated that PCB 126 possesses a potent anti-estrogenic activity in the sea bass in vivo as it inhibited the E2-induced VTG synthesis with an IC50 of 28 microg/kg. The results of this study suggest that the exposure of fish to xenoestrogens or anti-estrogens may alter, in addition to various physiological processes, the expression of specific CYPs and phase 2 enzymes, thereby reducing the capability of their detoxication system. PMID- 16219371 TI - Roles for mitochondria in pentamidine susceptibility and resistance in Leishmania donovani. AB - Pentamidine resistant Leishmania donovani was raised in the laboratory by stepwise exposure to increasing drug pressure until a line capable of growth in 8 microM pentamidine (R8) had been selected. An IC(50) value of 40 microM was determined for this line, some 50-fold higher than that recorded for the parental wild-type line. The pentamidine resistant promastigotes were cross-resistant to other toxic diamidine derivatives but not to antimonials or substrates of multidrug resistance pumps. Decreased mitochondrial transmembrane potential was observed in pentamidine resistant promastigotes. A substantial net decrease in accumulation of [(3)H]-pentamidine accompanied the resistance phenotype. Inhibitors of P-glycoprotein pumps, including prochlorperazine and trifluoperazine, did not reverse this decreased drug uptake, which distinguishes the L. donovani resistant line studied here from L. mexicana promastigotes previously studied for pentamidine resistance. Kinetic analysis identified a carrier with an apparent K(m) value of 6 microM for pentamidine. No significant difference between wild-type and resistant parasites could be detected with respect to this transporter in rapid uptake experiments. However, in longer-term uptake experiments and also using concentrations of pentamidine up to 1mM, it was demonstrated that wild-type cells, but not resistant cells, could continue to accumulate pentamidine after apparent saturation via the measured transporter had been reached. Agents that diminish the mitochondrial membrane potential inhibited this secondary route. A fluorescent analogue of pentamidine, 2,5-bis-(4 amidophenyl)-3,4-dimethylfuran (DB99), accumulated in the kinetoplast of wild type but not resistant parasites indicating that uptake of this cationic compound into mitochondria of wild-type cells was more pronounced than in the resistant line. These data together indicate that resistance to pentamidine in L. donovani is associated with alterations to the mitochondria of the parasites, which lead to reduced accumulation of drug. PMID- 16219372 TI - Characterization of Entamoeba histolytica alpha-actinin. AB - We have cloned, expressed and characterized a alpha-actinin-like protein of Entamoeba histolytica. Analysis of the primary structure reveals that the essential domains of the alpha-actinin protein family are conserved: an N terminus actin-binding domain, a C-terminus calcium-binding domain and a central helical rod domain. However, the rod domain of this Entamoeba protein is considerably shorter than the rod domain in alpha-actinins of higher organisms. The cloned Entamoeba 63 kDa protein is recognized by conventional alpha-actinin antibodies as well as binds and cross-links filamentous actin and calcium ions in the same manner as alpha-actinins. Despite the shorter rod domain this protein has conserved the most important functions of alpha-actinins. Therefore, it is suggested that this 63 kDa protein is an atypical and ancestral alpha-actinin. PMID- 16219373 TI - Analysis of spliceosomal complexes in Trypanosoma brucei and silencing of two splicing factors Prp31 and Prp43. AB - In trypanosomatids all mRNAs undergo trans-splicing, whereas cis-splicing is restricted to a few transcripts. Trans-splicing is mechanistically similar to cis splicing, however, little is known about the trans-splicing machinery and its underlying mechanism. In this study, we examined the involvement of splicing factors in cis- and trans-splicing by RNA interference (RNAi). Two factors (Prp31 and Prp43) were found to be essential for both pathways, suggesting that splicing factors are shared by these two reactions. We identified a 45S complex carrying pre-mRNA and all the U-snRNAs, including U1 and the SL RNA, suggesting that a single spliceosomal complex may potentially conduct both trans- and cis-splicing. PMID- 16219374 TI - Alterations in aortic vascular reactivity to angiotensin 1-7 in 17-beta-estradiol treated female SD rats. AB - Estrogen's suggested cardio-protective effects have come into question following the results of recent clinical trials. Two major components of the renin angiotensin system (RAS) that are modulated by estrogen are angiotensin converting enzyme, and the angiotensin II type 1 receptor. Further research has revealed several new components of the RAS, including angiotensin converting enzyme 2, its peptide product angiotensin 1-7 (Ang 1-7), and that peptide's receptor, Mas. These components appear to oppose the classical effects of the RAS, and may act to buffer the RAS in vivo. Recent work has shown that during pregnancy, when estradiol levels are elevated, renal and urinary Ang 1-7 are greatly increased. This study examined the effects of estradiol on the efficacy of Ang 1-7 in the rat aorta. Female Sprague-Dawley rats were ovariectomized and a subgroup was chronically treated with subcutaneous pellets of estradiol for 3 weeks. Thoracic aortas were harvested for assessment of in vitro vascular reactivity to Ang 1-7. The results demonstrated that increased estradiol exposure attenuated the relaxation response to Ang 1-7 in a dose-dependent manner. These findings are in contrast to recent work showing potentiated responses to Ang 1-7 in mesenteric arteries from estrogen-manipulated rats, and may suggest a regional specificity in estradiol-mediated changes in the RAS. PMID- 16219375 TI - Differences in higher-level functional capacity between participants and non participants in health checkups among the elderly. AB - The present study was conducted to characterize the lifestyles and health status of non-participants and to investigate whether diminished higher-level functional capacity may cause selection bias in non-compulsory mass health screening for the elderly. Using a self-administered questionnaire for evaluating the Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG Index of Competence) which consists of three sublevels, namely, instrumental self maintenance, intellectual activity and social role, we conducted a survey of 1543 (mean age, 64.9+/-12.5 years, 677 males and 866 females) out of all the 1701 individuals over the age of 40 residing in a village where mass health screening is conducted annually. The mean TMIG Index of Competence score was the highest in Group V (composed of 434 individuals who participated in the mass health screening conducted by the village), followed, in that order, by the score in Group W (composed of 531 individuals who had undergone a health checkup organized at their workplaces or by their family physicians, but not the one conducted by the village, during the previous year) and that in Group N (composed of 578 individuals who had not undergone any health checkup during the previous year). Group N showed a significantly lower mean TMIG Index of Competence score than Groups V and W. In regard to the scores for the sublevels of the index, Group N had a significantly lower percentage of subjects, both men and women, with perfect scores than Group V for all the sublevels, and also a significantly lower percentage of subjects with a perfect score for the intellectual activity than Group W. However, there were no significant differences in the percentages of subjects habituated to exercise, drinking or smoking among the three groups. Thus, special attention may need to be paid to selection bias in mass health screenings caused by differences in the higher-level functional capacity. PMID- 16219376 TI - Comparison between agarose gel electrophoresis and capillary electrophoresis for variable numbers of tandem repeat typing of Mycobacterium tuberculosis. AB - Variable numbers of tandem repeat (VNTR) typing of Mycobacterium tuberculosis was performed on 54 strains including 23 strains derived from 9 outbreaks. PCR amplicon sizes of 12 mycobacterial interspersed repetitive unit tandem repeat loci were measured using both agarose gel electrophoresis and capillary electrophoresis. Similarities using agarose gel electrophoresis of Euclidian distances among the 23 strains derived from the 9 outbreaks were significantly lower than that using capillary electrophoresis (Wilcoxon signed ranks test, P < 0.01). By clustering analysis using unweighted pair group method using arithmetic averages, all of the 23 strains derived from the 9 outbreaks were each clustered with more than 90% similarities based on the distance using capillary electrophoresis. In contrast, differential clusters with more than 90% similarity were observed with only 7 strains derived from 3 outbreaks when analyzed by agarose gel electrophoresis. These results indicated that measurement of PCR amplicon size of tandem repeat loci should be carried out using capillary electrophoresis and that agarose gel electrophoresis is not suitable for clustering analysis of M. tuberculosis VNTR typing. PMID- 16219377 TI - Postsynaptic receptor mechanisms underlying developmental speeding of synaptic transmission. AB - As animals mature the decay of postsynaptic currents become faster at a variety of synapses. This change is thought to contribute to a refinement of motor co ordination and to an increase in the precision of sensory perception and cognition. At cholinergic neuromuscular synapses and glycinergic and GABAergic inhibitory synapses, the developmental speeding of synaptic currents depends upon switches of receptor subunits and an ensuing acceleration in the kinetics of channel gating. At glutamatergic excitatory synapses, speeding in the decay time of NMDA receptor (NMDAR)-mediated excitatory postsynaptic currents (NMDA-EPSCs) is also dependent on developmental switches in NMDAR subunits. However, developmental speeding in the kinetics of AMPA receptor (AMPAR)-mediated EPSCs (AMPA-EPSCs) is caused by multiple factors. The decay time of AMPA-EPSCs can be shaped by the kinetics of channel gating or desensitization of AMPA receptors, depending upon the speed of transmitter clearance from the synaptic cleft. During postnatal development AMPAR channel gating and desensitization as well as the transmitter clearance speed up in kinetics. Given that the developmental speeding of synaptic currents play critical roles in the maturation of sensory and motor functions, any defect in this mechanism may seriously affect neuronal function. PMID- 16219378 TI - Effects of electrolyzed oxidizing water on reducing Listeria monocytogenes contamination on seafood processing surfaces. AB - The effects of electrolyzed oxidizing (EO) water on reducing Listeria monocytogenes contamination on seafood processing surfaces were studied. Chips (5 x 5 cm(2)) of stainless steel sheet (SS), ceramic tile (CT), and floor tile (FT) with and without crabmeat residue on the surface were inoculated with L. monocytogenes and soaked in tap or EO water for 5 min. Viable cells of L. monocytogenes were detected on all chip surfaces with or without crabmeat residue after being held at room temperature for 1 h. Soaking contaminated chips in tap water resulted in small-degree reductions of the organism (0.40-0.66 log cfu/chip on clean surfaces and 0.78-1.33 log cfu/chip on dirty surfaces). Treatments of EO water significantly (p<0.05) reduced L. monocytogenes on clean surfaces (3.73 log on SS, 4.24 log on CT, and 5.12 log on FT). Presence of crabmeat residue on chip surfaces reduced the effectiveness of EO water on inactivating Listeria cells. However, treatments of EO water also resulted in significant reductions of L. monocytogenes on dirty surfaces (2.33 log on SS and CT and 1.52 log on FT) when compared with tap water treatments. The antimicrobial activity of EO water was positively correlated with its chlorine content. High oxidation-reduction potential (ORP) of EO water also contributed significantly to its antimicrobial activity against L. monocytogenes. EO water was more effective than chlorine water on inactivating L. monocytogenes on surfaces and could be used as a chlorine alternative for sanitation purpose. Application of EO water following a thorough cleaning process could greatly reduce L. monocytogenes contamination in seafood processing environments. PMID- 16219379 TI - Promotion of hyphal growth and underlying chemical changes in Antrodia camphorata by host factors from Cinnamomum camphora. AB - The aim of this research was to investigate the hyphal growth-promoting factors (HGFs) of Antrodia camphorata from the host-related species, Cinnamomum camphora (CC) and the underlying chemical produced. The HGF was identified in the polysaccharide fraction of CC at levels ranging from 80 to 320 mg L(-1), and it maximally stimulated growth to 5.50 g L(-1) during a 14-day culture period compared to that of the control of 2.88 g L(-1). We also investigated the nature and chemical composition of the CC polysaccharide. Herein, size-exclusion column chromatography followed by high-performance anion-exchange chromatography after complete hydrolysis of the CC polysaccharide was performed to derive its molecular weight and sugar composition. The Mw values of the CC polysaccharide were determined to be 728.2, 187.5, 28.7, 7.5, and 1.9 kDa. Compositional analysis of the CC polysaccharide showed that galactosamine, mannose, and glucose were the major monosaccharides. Time-course studies of mycelial extracts of cultures revealed that prolonged incubation with the water-soluble extracts of CC resulted in an increase in the relative amounts of two lanostane-type compounds, i.e., dehydrosulphurenic acid and 15alpha-acetyl-dehydrosulphurenic acid, which are found in the fruiting bodies of A. camphorata. This finding offers the possibility of the reliable production of this medicinal fungus under laboratory conditions compared to its limited slow growth in nature. PMID- 16219380 TI - Genetic network driven control of PHBV copolymer composition. AB - We developed a detailed mathematical model describing the coupling between the molecular weight distribution dynamics of poly(3-hydroxybutyrate-co 3hydroxyvalerate) (PHBV) copolymer chains with those of hydroxybutyrate (HB) and hydroxyvalerate (HV) monomer formation. Sensitivity analysis of the model revealed that both the monomer composition and the molecular weight distribution of the copolymer chains are strongly affected by the ratio between the rates at which the two-monomer units are incorporated into the chains. This ratio depends on the relative HB and HV availability, which in turn is a function of the expression levels of genes encoding enzymes that catalyze monomer formation. Regulation of gene expression was accomplished through the aid of an artificial genetic network, the patterns of expression of which can be controlled by appropriately tuning the concentration of an extracellular inducer. Extensive simulations were used to study the effects of operating conditions and parameter uncertainties on the range of achievable copolymer compositions. Since the predicted conditions fell in the range of feasible bioprocessing manipulations, it is expected that such strategy could be successfully employed. Thus, the presented model constitutes a powerful tool for designing genetic networks that can drive the formation of PHBV copolymer structures with desirable characteristics. PMID- 16219381 TI - A multilayer membrane amperometric glucose sensor fabricated using planar techniques for large-scale production. AB - This paper reports on a multilayer membrane amperometric glucose sensor fabricated using planar techniques. It is characterized by good reproducibility and suitable for large-scale production. The glucose sensor has 82 electrode sets formed on a single glass substrate, each with a platinum working electrode (WE), a platinum counter electrode (CE) and an Ag/AgCl reference electrode (RE). The electrode sets are coated with a membrane consisting of five layers: gamma aminopropyltriethoxysilane (gamma-APTES), Nafion, glucose oxidase (GOX), gamma APTES and perfluorocarbon polymer (PFCP), in that order. Tests have shown that the sensor has acceptably low dispersion (relative standard deviation, R.S.D.=42.9%, n=82), a wide measurement range (1.11-111 mM) and measurement stability over a 27-day period. Measurements of the glucose concentration in a control human urine sample demonstrated that the sensor has very low dispersion (R.S.D.=2.49%, n=10). PMID- 16219382 TI - Immunohistochemical examination of the role of Fas ligand and lymphocytes in the pathogenesis of human liver yellow fever. AB - Yellow fever is an infectious, non-contagious disease caused by an RNA virus of the family Flaviviridae, which is transmitted to man by the bite of hematophagous mosquitoes. Infection with the yellow fever virus can progress with lesions in the heart, kidneys, central nervous system, and liver. In the liver, the histopathological picture is characterized by necrosis, steatosis and hepatocyte apoptosis, with a preferential midzone distribution. In the present study, liver samples from fatal patients with yellow fever were analyzed. The histopathological pattern was characterized by steatosis, lytic necrosis and hepatocyte apoptosis associated with a moderate mononuclear inflammatory infiltrate. The inflammatory component mainly consisted of CD4+ T lymphocytes, followed by CD8+ T lymphocytes, which showed a preferential portal and midzone distribution. Immunoreactivity to Fas ligand was mainly observed in hepatocytes of the midzone region. Based on these findings, we conclude that lymphocytes play an important role in the genesis of hepatic lesions in severe yellow fever, inducing hepatocyte apoptosis through the binding to Fas receptors. However, further studies are necessary to investigate the participation of other immune factors and to quantify the role of the cytotoxic cellular response in the lesion evolution during the course of disease in the liver. PMID- 16219383 TI - Ultrasound enhances in vivo tumor expression of plasmid DNA by PEG-introduced cationized dextran. AB - This study is an investigation to experimentally confirm whether or not ultrasound (US) irradiation is effective in enhancing the in vivo gene expression of plasmid DNA in tumor. Dextran was cationized by introducing spermine to the hydroxyl groups to allow to polyionically complex with a plasmid DNA. The cationized dextran prepared was additionally modified with poly(ethylene glycol) (PEG) molecules which have an active ester and methoxy groups at each terminal, to obtain cationized dextran with different percentages of PEG introduced. Various cationized dextrans with or without PEG introduction were mixed with a plasmid DNA of LacZ to form cationized dextran-plasmid DNA complexes. Electrophoretical examination revealed that the plasmid DNA was complexed both with the cationized dextran and PEG-introduced cationized dextran, irrespective of the PEG introduction percentage, although the higher N/P ratio was needed for plasmid DNA complexation with the latter. By complexation with the cationized dextran, the zeta potential of plasmid DNA was changed to be positive. The charge of PEG-introduced cationized dextran-plasmid DNA complexes became close to 0 mV as their percentage of PEG introduced increased, although the molecular size was about 250 nm, irrespective of the PEG introduction. When cationized dextran plasmid DNA complexes with or without PEG introduction were intravenously injected to mice carrying a subcutaneous Meth-AR-1 fibrosarcoma mass and the subsequent US irradiation to the tumor mass percutaneously, the PEG-introduced cationized dextran-plasmid DNA complex plus US irradiation enhanced the tumor level of gene expression to a significantly high extent compared with the cationized dextran-plasmid DNA complex and free plasmid DNA with or without US irradiation. The enhanced level depended on the time period and timing of US irradiation. Fluorescent microscopic studies revealed that the localization of plasmid DNA and the gene expression were observed in the tumor tissue injected with the PEG-introduced cationized dextran-plasmid DNA complex plus the subsequent US irradiation. We conclude that complexation with the PEG-introduced cationized dextran combined with US irradiation is a promising way to target the plasmid DNA to the tumor for gene expression. PMID- 16219384 TI - Linear double-stranded DNA that mimics an infective tail of virus genome to enhance transfection. AB - Our previous work showed that a natural beta-(1-->3)-d-glucan schizophyllan (SPG) can form a stable complex with single-stranded oligonucleotides (ssODNs). When protein transduction peptides were attached to SPG and this modified SPG was complexed with ssODNs, the resultant complex could induce cellular transfection of the bound ODNs, without producing serious cytotoxicity. However, no technique was available to transfect double-stranded DNAs (dsDNA) or plasmid DNA using SPG. This paper presents a new approach to transfect dsDNA, showing preparation and transfection efficiency for a minimal-size gene having a loop-shaped poly(dA)(80) on both ends. This poly(dA) loops of dsDNA can form a complex with SPG. An siRNA coding dsDNA with the poly(dA) loop was complexed with Tat-attached SPG to silence luciferase expression. When LTR-Luc-HeLa cells that can express luciferase under the control of the LTR promoter were exposed to this complex, the expression of luciferase was suppressed (i.e., RNAi effect was enhanced). Cytotoxicity studies showed that the Tat-SPG complex induced much less cell death compared to polyethylenimine, indicating that the proposed method caused less harm than the conventional method. The Tat-SPG/poly(dA) looped dsDNA complex had a structure similar to the viral genome in that the dsDNA ends were able to induce transfection and protection. The present work identifies the SPG and poly(dA) looped minimum-sized gene combination as a candidate for a non-toxic gene delivery system. PMID- 16219385 TI - Entropies for detection of epilepsy in EEG. AB - The electroencephalogram (EEG) is a representative signal containing information about the condition of the brain. The shape of the wave may contain useful information about the state of the brain. However, the human observer cannot directly monitor these subtle details. Besides, since bio-signals are highly subjective, the symptoms may appear at random in the time scale. Therefore, the EEG signal parameters, extracted and analyzed using computers, are highly useful in diagnostics. The aim of this work is to compare the different entropy estimators when applied to EEG data from normal and epileptic subjects. The results obtained indicate that entropy estimators can distinguish normal and epileptic EEG data with more than 95% confidence (using t-test). The classification ability of the entropy measures is tested using ANFIS classifier. The results are promising and a classification accuracy of about 90% is achieved. PMID- 16219386 TI - Development of a simulation program for estimating hospital incomes under the prospective payment system. AB - A prospective payment system based on diagnosis procedure combination (DPC/PPS) was introduced to acute care hospitals in Japan in April 2003. In order to increase hospital income, hospitals must shorten the average length of stay (ALOS) and increase the number of patients. We constructed a simulation program for evaluating the relationships among ALOS, bed occupation rate (BOR) and hospital income of hospitals in which DPC/PPS has been introduced. This program can precisely evaluate the hospital income by regulating the ALOS and the number of patients for each DPC. By using this program, it is possible to predict the optimum ALOS and optimum number of inpatients for each DPC in order to increase hospital income. PMID- 16219387 TI - Short chain fatty acids regulate tyrosine hydroxylase gene expression through a cAMP-dependent signaling pathway. AB - Multiple intracellular and extracellular regulatory factors affect transcription of the tyrosine hydroxylase (TH) gene encoding the rate-limiting enzyme in the biosynthesis of the neurotransmitters dopamine, norepinephrine and epinephrine. Short chain fatty acids like butyrate are known to alter TH gene expression, but the mechanism of action is unknown. In this report, transient transfection assays identified the proximal TH promoter to contain sufficient genetic information to confer butyrate responsiveness to a reporter gene. Deletion studies and gel shift analyses revealed that the promoter region spanning the cAMP response element is an absolute requirement for transcriptional activation by butyrate. The branched short chain fatty acid valproate is used for seizure control in humans. Significantly, it has a similar aliphatic structure to butyrate, and it was found to have similar effects on TH in PC12 cells. Site-directed mutagenesis indicated that the effects of both fatty acids were mediated through the canonical CRE. Butyrate treatment also resulted in CREB phosphorylation without changing CREB protein levels. The increased phosphorylation of CREB correlated with accumulation of TH mRNA. The adenylate cyclase inhibitor dideoxyadenosine blocked both CREB phosphorylation and accumulation of TH mRNA. The data are consistent with the conclusion that butyrate induces post-translational modifications of pre existing CREB molecules in a cAMP/PKA-dependent manner to alter TH transcription. These results support the role of butyrate as a novel exogenous regulatory factor in TH gene expression. Our data delineate a molecular mechanism through which diet-derived environmental signals (e.g. butyrate) can modulate catecholaminergic systems by affecting TH gene transcription. PMID- 16219389 TI - T-817MA, a novel neurotrophic agent, improves sodium nitroprusside-induced mitochondrial dysfunction in cortical neurons. AB - 1-{3-[2-(1-Benzothiophen-5-yl)ethoxy]propyl}-3-azetidinol maleate (T-817MA), a novel neurotrophic agent, protects against amyloid-beta peptide- or hydrogen peroxide-induced neuronal death. The exact mechanism of the neuroprotection is not known. This study examines the effects of T-817MA on oxidative stress-induced cytotoxicity in primary rat cortical neurons. Treatment with the NO donor sodium nitoroprusside (SNP) at 300microM decreased cell viability and induced apoptotic cell death. SNP-induced neuronal toxicity was accompanied by a decrease in mitochondrial transmembrane potential without an increase in the expression of CHOP and GRP78 mRNAs, endoplasmic reticulum stress makers. T-817MA at 0.1 and 1microM attenuated the neurotoxicity in a dose-dependent way and the protective effect required pretreatment for more than 8h. T-817MA attenuated SNP-induced decrease in mitochondrial transmembrane potential. In addition, the agent reduced SNP-induced increase in mitochondrial reactive oxygen species (ROS) production. The effects of T-817MA on SNP-induced decrease in cell viability and SNP-induced increase in mitochondrial ROS production were blocked by cycloheximide. These results suggest that T-817MA improves SNP-induced mitochondrial dysfunction in cortical neurons in a newly synthesized protein-mediated mechanism and this effect contributes to its neuroprotective effect. PMID- 16219388 TI - Altered long-term synaptic plasticity and kainate-induced Ca2+ transients in the substantia gelatinosa neurons in GLU(K6)-deficient mice. AB - Functional kainate receptors are expressed in the spinal cord substantia gelatinosa region, and their activation contributes to bi-directional regulation of excitatory synaptic transmission at primary afferent synapses with spinal cord substantia gelatinosa neurons. However, no study has reported a role(s) for kainate receptor subtypes in long-term synaptic plasticity phenomena in this region. Using gene-targeted mice lacking glutamate receptor 5 (GLU(K5)) or GLU(K6) subunit, we here show that GLU(K6) subunit, but not GLU(K5) subunit, is involved in the induction of long-term potentiation of excitatory postsynaptic potentials, evoked by two different protocols: (1) high-frequency primary afferent stimulation (100 Hz, 3 s) and (2) low-frequency spike-timing stimulation (1 Hz, 200 pulses). In addition, GLU(K6) subunit plays an important role in the expression of kainate-induced Ca2+ transients in the substantia gelatinosa. On the other hand, genetic deletion of GLU(K5) or GLU(K6) subunit does not prevent the induction of long-term depression. These results indicate that unique expression of kainate receptors subunits is important in regulating spinal synaptic plasticity and thereby processing of sensory information, including pain. PMID- 16219390 TI - Medial temporal lobe volume of nondemented elderly individuals with poor cognitive functions. AB - Poor cognitive performance of elderly individuals with low educational attainment is often difficult to interpret in dementia evaluation. Lack of education, as well as dementia, is often associated with poor cognitive test performance. To elucidate the underlying structural change of low cognitive performance in elderly individuals with low educational attainment, this study examined the relationship between low cognitive performance (LCP) and brain volumes, especially regions vulnerable to Alzheimer's disease, in nondemented elderly Koreans. Individuals with LCP (n=14) were matched on age and education with individuals with normal cognitive performance (n=14). The two groups were compared on the MR-based volumetric measures in the hippocampus, the entorhinal cortex, the amygdala, the temporal lobe, the frontal lobe, the cerebrum, and the intracranial cavity. Intracranial volume (p<.05) and absolute hippocampus (p<.05) and frontal lobe volumes (p<.05) were significantly reduced in individuals with LCP. Normalized volumes of the hippocampus and the frontal lobe did not differ in the two cognitive performance groups. ICV was associated with the K-DRS scores. General cognitive functioning of the LCP individuals, measured with the Korean version of the DRS, did not deteriorate in the 1- or 2-year follow-up cognitive tests. LCP in a nondemented elderly population with limited education appears to be associated with stable lower intelligence rather than increased risk for dementia of the Alzheimer's type. PMID- 16219391 TI - Studies of the effects of high fat diets on cognitive function in a rat model. AB - This research summarises a research program that is concerned with the effects of high fat diets on cognitive function in rats. The diets selected accurately represent current upper limits of human fat consumption in western societies. Rats fed with diets high in saturated or unsaturated fat for 3 months, were severely impaired on a range of learning and memory tasks. Related studies showed that these effects were modulated by concentration of fat, environmental influences, and treatment with glucose. More work is needed to identify the neurobiological mechanisms underlying this impairment but saturated fatty acid intake, as well as insulin resistance and glucose intolerance may be important factors. In demonstrating a clear relationship between obesity and cognitive impairment, this research has important implications for aging. Older adults are particularly vulnerable to the adverse effects of dietary fat and the present results underscore the importance of evidence that seniors with marginal levels of nutrient intake often perform poorly on tests of cognitive function, and are at increased risk for various forms of dementia. PMID- 16219392 TI - Effects of multiple pulmonary aspirations of enteral solutions on lung tissue damage. AB - BACKGROUND & AIM: The aim of this study was to compare the pulmonary histopathologic effects of enteral solutions with various lipid content, after multiple aspirations in rats. METHODS: Thirty Wistar albino rats were randomly assigned to one of three groups (n = 10). Saline solution (0.9%) (group C, control), Impact (lipid content, 28 g/l; group I), Pulmocare (lipid content, 93.3g/l; group P) were injected into the lung through the trachea, in a volume of 0.8 ml/kg. The aspiration procedure was performed three times in total, in every 2 days. After seven days from the first aspiration, rats were killed, and lungs were examined for histopathologic examination. RESULTS: Alveolar histiocytes were statistically higher in left lungs of the group I than the left ones of the control group (P < 0.05). Lipid-laden alveolar macrophages were significantly higher in left lungs of groups I and, P than left lungs of the control group (P<0.05). CONCLUSION: Lung tissue damage occurring after multiple pulmonary aspirations of Impact and Pulmocare, is histopathologically similar to each other, and is in the form of lipoid pneumonia. In cases of multiple pulmonary aspirations, volume of the aspirate and chronicity of the aspiration look like major impact factors rather than the amount of the lipid. PMID- 16219393 TI - The prevalence of malnutrition in hospitals can be reduced: results from three consecutive cross-sectional studies. AB - AIMS: To assess and compare the rates of malnutrition in Hammersmith Hospital NHS Trust over a 5-year period following changes in hospital nutrition care strategies. METHODS: DESIGN: Three consecutive cross-sectional studies carried out in 1998, 2000 and 2003. SETTING: Inpatients at Hammersmith Hospital NHS Trust. PARTICIPANTS: A total of 2283 inpatients aged over 16 years old, 686 in 1998, 780 in 2000, 817 in 2003. Inpatients excluded: ventilated patients, ante/post-natal women and people aged <16. INTERVENTIONS: Improvements in the catering service and nutrition education provision in 2000, and the implementation of a nutrition screening tool and 'Better Hospital Food' in 2003. MAIN OUTCOME MEASURE: Prevalence of malnutrition. RESULTS: There was a reduction in the prevalence of malnutrition in 2000 and 2003 from baseline data in 1998 (1998: 23.5%, 161/686; 2000: 20.4%, 159/780; 2003: 19.1%, 156/817; P<0.001). The odds ratio of being either at risk of malnutrition or malnourished was reduced in both 2000 and 2003 by approximately 33% (P=0.001). Indicators of good nutritional practice also improved: Weighing patients on admission increased from 37.5% (257/686) in 1998, to 42.9% (335/780) in 2000, and 59.6% (487/817) in 2003 (P0.001). Dietetic referrals also increased from 31.5% (216/686) in 1998 to 41.6% (340/817) in 2003 (P<0.001)(no change in 2000, 31%, 242/780). Appropriate referrals also improved, results showing that the proportion of malnourished patients who were referred showed a dramatic increase in 2003 (1998: 91/161, 56.5%; 2000: 85/159, 53.5%; 2003: 111/156, 71.2%; P=0.003). CONCLUSIONS: The prevalence of malnutrition in hospital can be influenced by the implementation of a variety of nutrition care strategies, which target identification of malnutrition and its treatment. PMID- 16219394 TI - Assessment of measles incidence, measles-related complications and hospitalisations during an outbreak in a southern Italian region. AB - A large measles epidemic occurred in 2002 in Campania, a region of southern Italy with inadequate vaccination coverage. We evaluated the burden of the outbreak in children <15 years of age using different data sources. The measles standardized incidence rate was 5,757/100,000, corresponding to 63,368 estimated cases (95% CI: 59,544--67,373). Measles virus strains were identified as belonging to the D7 genotype. The estimated complication rate was 7.6%. A total of 972 measles hospitalisations were detected, giving a hospitalisation rate of 88.3/100,000. Three deaths occurred. These results show that measles can still represent a serious health threat even in industrialized countries. PMID- 16219395 TI - Modulating the adjuvanticity of alum by co-administration of muramyl di-peptide (MDP) or Quil-A. AB - The characterization of the immunological cascades of the innate immune system activated by pathogen associated molecular patterns (PAMP) recognized by pattern recognition receptors (PRR) have allowed the elucidation of the mechanisms underlying the immunomodulatory properties of adjuvants. Thus, the combinatorial use of adjuvants with specific, complementary functions is investigated to achieve tailored immune responses to subunit vaccines. We have previously shown how combinatorial administration of chitosan and cholera toxin B or muramyl-di peptide (MDP) intranasally, but not intramuscularly, can allow small doses of MDP which, when administered alone cannot adjuvantise Helicobacter pylori urease (rUre), achieve an immunomodulatory effect through the specific physiological effect of chitosan. The aim of this study was to investigate if in the context of rUre the adjuvantising effect of MDP could be realized via the intramuscular route by combination with aluminium hydroxide, as compared with the routinely used veterinary adjuvant combination of alum and Quil-A. Serum IgG kinetics were comparable between the two adjuvant combination groups. However, the alum + MDP combination afforded higher antigen-specific recall responses in splenocyte cultures, associated with elevated release of the type I immune response cytokines IFN-gamma and IL-2. This data suggests that the adjuvanticity of MDP can be modulated in the context of alum in a manner dissimilar to that of Quil-A, achieving a balancing effect on the responses elicited by alum adjuvantisation. PMID- 16219397 TI - The immunogenicity and protective efficacy of Mtb8.4/hIL-12 chimeric gene vaccine. AB - DNA vaccination has emerged as a powerful approach in the search for a more efficacious vaccine against tuberculosis (TB). In this study, we evaluated the immunogenicity and protective efficacy of Mtb8.4/hIL-12 chimeric gene vaccine. The Mtb8.4/hIL-12 chimeric gene was amplified by PCR and cloned into the eukaryotic expression vector pCI-neo. C57BL/6N mice were vaccinated with Mtb8.4/hIL-12 chimeric gene vaccine for three times at 3 weeks intervals. Four weeks after the final inoculation, three mice per group were sacrificed to assess cytokine response and CTL induction and the other five mice per group were challenged intravenously in a lateral tail vein with 1 x 10(6) CFU of virulent Mycobacterium tuberculosis H37Rv. Spleen and the left lung were harvested from each mouse at 4 weeks after infection and homogenized in sterile saline. Serial dilutions of organ homogenates were plated on L-J agar and incubated 37 degrees C until colonies were visible 4 weeks later. Protective efficacies in each experiment were expressed as reduced CFU and were compared with the negative control group. The right lung was obtained from each mouse and immediately inflated with and stored in 10% formalin saline. Tissues were embedded in paraffin, sectioned and stained with hematoxylin and eosin. Mtb8.4/hIL-12 chimeric gene vaccine induced the secretion of more of Th1 cytokines, but not IL 4 and enhanced CTL activity. Mice immunized with Mtb8.4/hIL-12 chimeric gene vaccine had fewer and smaller tubercles than control groups. As expected, control mice had the highest bacterial loads in both lung and spleen. Immunization with Mtb8.4/hIL-12 chimeric gene vaccine could remarkably reduced CFU counts in organs. When it was used to construct the chimeric gene vaccine, hIL-12 could improve the immune efficacy of Mtb8.4 gene vaccine. PMID- 16219398 TI - Immune responses to human papillomavirus. AB - The immune system uses innate and adaptive immunity to recognize and combat foreign agents that invade the body, but these methods are sometimes ineffective against human papillomavirus (HPV). HPV has several mechanisms for avoiding the immune system. HPV infects, and multiplies in keratinocytes, which are distant from immune centers and have a naturally short lifespan. The naturally short life cycle of the keratinocyte circumvents the need for the virus to destroy the cell, which would trigger inflammation and immune response. In addition, HPV downregulates the expression of interferon genes. Despite viral immune evasion, the immune system effectively repels most HPV infections, and is associated with strong localized cell mediated immune responses. New prophylactic L1 virus-like protein vaccines for HPV 16 and 18 and HPV 6, 11, 16, and 18 are in phase 3 trials. Available data suggests that these vaccines are safe, produce high levels of antibodies, and are effective at preventing HPV infection. PMID- 16219399 TI - Definitive toxicology and biodistribution study of a polyvalent DNA prime/protein boost human immunodeficiency virus type 1 (HIV-1) vaccine in rabbits. AB - A toxicity and immunogenicity study, evaluating the safety of a polyvalent DNA prime/protein boost HIV-1 vaccine (DP6-001), was examined in rabbits. Animals were primed with a cocktail of six different DNA plasmids expressing five HIV-1 env genes and one gag gene followed by boosting with five gp120 proteins homologous to the DNA vaccines. The vaccine was shown to be immunogenic as evident from the induction of high-titered anti-Env and anti-Gag antibodies. There was an absence of detectable adverse effects on key toxicology parameters. Although plasmids persisted in the injection sites following single administration for 64 days, no evidence of integration into the host genomic DNA was observed. These studies demonstrate that a novel polyvalent DNA prime/protein boost vaccine lacks signs of toxicity and DNA integration in a rabbit model, and immunogenicity and toxicology data support clinical testing of the vaccine in humans. PMID- 16219400 TI - Letter to the editor. PMID- 16219402 TI - Simulating the long-term chemistry of an upland UK catchment: heavy metals. AB - CHUM-AM was used to investigate the behaviours of atmospherically-deposited heavy metals (Ni, Cu, Zn, Cd and Pb) in three moorland sub-catchments in Cumbria UK. The principal processes controlling cationic metals are competitive partitioning to soil organic matter, chemical interactions in solution, and chemical weathering. Metal deposition histories were generated by combining measured data for the last 30 years with local lake sediment records. For Ni, Cu, Zn and Cd, default parameters for the interactions with organic matter provided reasonable agreement between simulated and observed present-day soil metal pools and average streamwater concentrations. However, for Pb, the soil binding affinity in the model had to be increased to match the observations. Simulations suggest that weakly-sorbing metals (Ni, Zn, Cd) will respond on timescales of decades to centuries to changes in metal inputs or acidification status. More strongly sorbing metals (Cu, Pb) will respond over centuries to millennia. PMID- 16219403 TI - Spatio-temporal and species-specific variation in PBDE levels/patterns in British Columbia's coastal waters. AB - Congener-specific levels of PBDEs were measured in the livers and some muscle tissues of Dungeness crab (Cancer magister), English sole (Pleuronectes vetulus) and spiny dogfish (Squalus acanthias). Highest concentrations (1,200-560 ng/g lipid) were found in crab collected near heavily urbanized areas (pop. approximately 0.3-1.8 million), followed by moderate levels at pulp/paper mills sites ( approximately 150 ng/g), and lowest levels occurred in areas that were somewhat removed from industrial/populated areas (< 24 ng/g). Temporal increases in total PBDEs and particularly in BDE-47 for Dungeness crab collected near pulp and paper and urbanized areas between 1994 and 2000 were observed. These correspond to Canadian and worldwide trends seen for PBDEs in biota. English sole and dogfish showed a pattern similar to that of the Columbia River whitefish samples, which corresponded closely to the patterns in the "penta" commercial mixture. Conversely, Dungeness crab were enriched in lower chlorinated PBDEs, particularly BDE-47 and BDE-49, compared to the fish and shark species from BC. PMID- 16219404 TI - Persistent organic pollutants and sedimentary organic matter properties: a case study in the Kishon River, Israel. AB - The Kishon River, the second largest coastal river in Israel, has been severely polluted for several decades. Sediments from upstream and downstream sites of the river were analyzed, lipid-extracted and evaluated for phenanthrene uptake. Total polycyclic aromatic hydrocarbon (PAH) concentration in downstream sediments was 299 microg kg(-1), consisting mostly of petrogenic-derived PAHs. Downstream sedimentary lipids were found to be dominated by fresh and decomposed petroleum derived n-alkanes. The total PAH concentration in upstream sediments was 173 microg kg(-1), consisting mostly of pyrogenic-derived PAHs, whereas lipids from these sediments were mostly vegetation-derived. Spectroscopic data suggested an exceptionally high aromatic content in downstream humic acid, which originated from PAHs attached to its structure. Sorption data suggested that upstream sedimentary cuticle-derived lipids function as a sorption domain, while downstream sedimentary lipids, consisting of shorter-chain-length petroleum derived alkanes, compete with phenanthrene for sorption sites. PMID- 16219405 TI - Influence of hydrology on heavy metal speciation and mobility in a Pb-Zn mine tailing. AB - Among the inorganic toxicants of greatest concern in mine tailings, Pb, Zn, Cu, Cd and As figure prominently due to their abundance and potential toxicity. Here we report on their biolability and solid-phase speciation in two sediment cores subject to variable hydrological regimes at an abandoned pyritic mine tailing. The oxic conditions of well-drained sediments induced pyrite oxidation and the subsequent liberation of H+, SO4(2-) and considerable quantities of Fe(III), which precipitated as goethite. Solubility of Pb, Zn, Cu and Cd was closely coupled to pH and goethite presence. Metal lability was particularly low in zones of neutralization, formed by the accumulation of calcite, first carried then deposited by percolating waters in both saturated and unsaturated cores. We conclude that differential hydrology induces variable heavy metal speciation and biolability in Pb-Zn mine tailings, and suggest that site-specific risk assessments must account for past and present hydrological regimes. PMID- 16219406 TI - Do individual and program factors matter in the utilization of maternal care services in rural India?: a theoretical approach. AB - Current studies on the utilization of maternal care services in India focus on individual factors. In the present study, we use the theoretical model developed by Andersen and Newman [1973. Societal and individual determinants of medical care utilization in the United States. Milbank Memorial Fund Quarterly, 51(1), 95 124] to understand the utilization of maternal care services in rural areas of India. This theoretical model uses individual and welfare program-or system factors-to study health care utilization. Data collected through the National Family Health Survey-2 are used in the present study. The results suggest that in addition to individual characteristics, program and system factors influence the utilization of maternal care in rural areas. Program factors, particularly educational activities promoting the benefits of maternal care services carried out through mahila mandal and anganwadi centers, are important in increasing the use of maternal care services in rural areas. Additionally, the results indicate that the mere presence of a private health care facility need not necessarily improve utilization. Increase in utilization is observed among households if the health worker visited these households during pregnancy. More state-specific studies incorporating both program and individual characteristics are recommended to further strengthen our understanding of the utilization of health care services in general and maternal care services specifically. PMID- 16219407 TI - Cerebral infarction following thrombolysis for massive pulmonary embolism. AB - A 29-year-old male developed a fatal stroke 6 h after successful thrombolysis for massive pulmonary embolism. Autopsy showed thrombus protruding through a patent foramen ovale (PFO). A strand of thrombus extended from the aortic arch into the left common carotid artery. The brain showed extensive infarction of the left fronto-parietal area. Thrombolysis caused initial disintegration of the embolism. It is likely that thrombolysis caused fragments of clot to later break lose and embolise into the cerebral circulation. We discuss the need for risk stratification in patients who present with massive pulmonary embolism and PFO. PMID- 16219408 TI - Attitudes to basic life support among medical students following the 2003 SARS outbreak in Hong Kong. AB - BACKGROUND: In 2003 severe acute respiratory syndrome (SARS) affected 1,755 people in Hong Kong, including 386 health care professionals, some of whom were infected during resuscitation attempts of affected patients. This study seeks to explore whether this epidemic has altered the willingness of Hong Kong medical students to perform basic life support and mouth-to-mouth ventilation during an out-of-hospital cardiac arrest. METHODS: A questionnaire was used to survey Year 4 medical students at the end of their undergraduate anaesthesia attachment, during which basic life support (BLS) skills were taught. The survey was conducted during July and August 2003, approximately two months after Hong Kong was removed from the World Health Organisation SARS Infected Areas list, and was designed to examine student confidence in BLS skills, their perceptions of the risks associated with performing BLS and their willingness to perform BLS in varying situations. RESULTS: The response rate was over 60% (35 from a possible 54). Students were positive regarding the adequacy of their BLS training. They were concerned about disease transmission during resuscitation but were less positive regarding whether the risks had increased due to SARS. In all situations they were significantly more likely to perform mouth-to-mouth ventilation for a family member compared with a stranger (p < 0.001) and to withhold mouth-to-mouth ventilation if either vomit or blood were present in the victim's mouth. CONCLUSIONS: Hong Kong medical students feel able to perform BLS if required. They are concerned about the risk of disease transmission, including SARS, during resuscitation, but would be more likely to withhold mouth-to-mouth resuscitation in the presence of vomit or blood than due to a fear of contracting SARS. PMID- 16219409 TI - Influence of resuscitation volume on blood cells TNF production in a murine model of haemorrhage. AB - The influence of haemorrhage and resuscitation on Tumour Necrosis Factor (TNF) production by whole blood cultures under endotoxin (Escherichia coli LPS) stimulation was investigated in male BALB/c mice. Haemorrhagic shock was induced by removing 0.026 +/- 0.003 mL of blood/g via a cardiac puncture, resulting in a 50% decrease in arterial pressure and a metabolic adidosis. Animals were resuscitated successfully (normotensive) despite a residual base deficit and hyperlactatemia, 60 min after the haemorrhage by the restitution of shed blood volume (SBV) with or without an additional volume of crystalloid (Lactated Ringer's solution) equal to 50, 100 (isovolumetric resuscitation) or 200% of SBV. Pulmonary failure (hypoxia-hypercarbia) and myocardial injury (troponin I release) was observed in this last group. TNF production by whole blood cultures stimulated ex vivo by LPS was estimated 60 min after the end of resuscitation. Haemorrhage resulted in a 48-60% decrease in TNF production. This decrease so called 'leukocyte deactivation' was not modified by the restitution of SBV with or without crystalloid except for isovolumetric resuscitation which resulted in the cytokine level returning to control in the absence of clear cardiopulmonary dysfunction. In the present murine model of haemorrhage, modifying resuscitation volume influences in vitro TNF production in whole blood cultures challenged by LPS. PMID- 16219410 TI - Acquired lymphangioma circumscriptum of the vulva mimicking genital warts. PMID- 16219411 TI - Effect of pesticides on estrogen receptor transactivation in vitro: a comparison of stable transfected MVLN and transient transfected MCF-7 cells. AB - The estrogenic potential of four pesticides (endosulfan, prochloraz, tolchlofos methyl and propamocarb) was compared in parallel with 17beta-estradiol (E2) by reporter constructs in transient transfected MCF-7BUS and in stable transfected MVLN cells. Similar detection limit and half maximum effect concentration was determined for E2, whereas the maximum effect concentration of E2 was much higher in MCF-7BUS (10 nM) than in MVLN (150 pM), with the induced response being approximately six times the level in MVLN cells. Alone the four pesticides elicited the same relative response in the two bioassays, and similar data was obtained upon co-exposure with E2 for endosulfan and propamocarb. In contrast to the transient MCF-7BUS system, endosulfan further increased the E2 induced response in MVLN cells, whereas propamocarb did not induce the E2 response in MVLN cells as observed in MCF-7BUS cells. In conclusion, high agreement between the two reporter assays was observed, although some performance characteristics have to be considered. PMID- 16219412 TI - Epigenetic control of ovarian function: the emerging role of histone modifications. AB - The dynamic nature of the ovarian follicle makes it an ideal model to study the coordinated activation and inactivation of genes related to cell growth and differentiation. Much progress has been made in identifying transcription factors that promote the transcription of ovarian genes mediating gonadotropin action and steroidogenesis, but how these factors promote transcription in the context of chromatin is not well understood. Over the past 5 years, epigenetic regulation of ovarian genes through histone modifications has been the focus of an increasing number of studies. Several coactivators and corepressors associated with transcription factors are in fact histone acetyltransferases and histone deacetylases mediating the hyperacetylation and hypoacetylation of histones, respectively. Hyperacetylation of lysine residues in the core histone tails promotes chromatin alterations that favor transcription, whereas hypoacetylation of histones promotes gene silencing or repression. Not only does the acetylation status of the core histones determine whether chromatin remodeling occurs, but histone phosphorylation and methylation may serve equally important roles. For example, the combination of histone H3 phosphorylation and acetylation concertedly favors transcription. In addition, specific lysine methylations (e.g., K9 of histone H3) repress gene expression whereas other methylations promote gene expression. It is most likely the combination of histone modification events that regulate the initiation of transcription. Understanding how ovarian hormones control specific histone modifications will help us understand how follicular cells can switch from active gene pools governing cell proliferation to those gene groups controlling terminal differentiation. Progress in elucidating the ovarian specific regulation of histone modifying enzymes as well as identification of their target gene pools at different stages of the follicular cycle is expected in the next few years. PMID- 16219413 TI - Current insights into the role of transforming growth factor-beta in bone resorption. AB - Transforming growth factor-beta (TGF-beta) elicits a variety of effects on cellular proliferation and differentiation. The major repository for TGF-beta is bone, where it possesses separate facilitative and suppressive actions on osteoclast differentiation and bone resorption. Without a direct enabling stimulus from TGF-beta monocytes cannot form osteoclasts but instead follow macrophage differentiation pathways. This facilitative action depends on an ability to promote a state in which precursors are resistant to anti-osteoclastic inflammatory signals. Following the initiation of resorption TGF-beta is released from bone matrix. This acts on osteoblasts to reduce the availability of the osteoclast differentiation factor, RANKL (receptor activator of NFkappaB ligand) and thereby indirectly limits further osteoclast formation. Thus TGF-beta has a fundamental role in the control of bone resorption having actions that first allow monocytes to develop into osteoclasts then subsequently limiting the extent and duration of resorption after its release from the bone matrix. PMID- 16219414 TI - Metabonomics and the endocrine system. AB - Proton-NMR-based metabonomics offers a rare opportunity as a definitive screening technique for biofluids and tissue biopsies. The procedure is extraordinary in that it allows the 'complete biochemical picture' to be examined at one time and is able to detect subtle but repeatedly consistent disparities that may be occurring in different, and perhaps unrelated, biochemical pathways. Such metabolic responses to an initial perturbation in homeostasis may be followed over a sequential time-course to their eventual dissipation or consequent sequelae. The application of this technique is beginning slowly to filter into the area of endocrine research and has been used to examine long-term and diffuse physiological alterations that may occur following such events as anabolic steroid treatment of cattle and the exposure of endometrial cells to tamoxifen. Although only modest inroads have been made so far, this technique promises immense potential for future researches within the endocrine field. PMID- 16219415 TI - MNU-induced carcinogenesis of rat mammary gland: effect of thyroid hormone on expression of retinoic acid receptors in tumours of mammary gland. AB - The rat model of N-methyl-N-nitrosourea (MNU)-induced mammary carcinomas is well established animal model for breast cancer. This study was carried out to investigate whether hypothyroid (thyroidectomy or PTU treatment) or hyperthyroid status of female rats would affect MNU-induced mammary carcinogenesis with specific focus on both retinoid and rexinoid receptor expression in mammary tumours. Application of PTU before and during MNU-induced mammary gland carcinogenesis yielded in a marked decrease of the number and volume of tumours per animal, however, there was no effect of hypothyroid state in thyroidectomized rats as well as hyperthyroid state concerning the number and volume of tumours. Mammary tumours of in euthyroid group of MNU animals showed that there was no tumour, in which all of subtypes of retinoid and rexinoid receptors were expressed. A different pattern of expression of retinoid or rexinoid receptors was found either in MNU-induced mammary carcinomas in both hypothyroid and hyperthyroid rats. PMID- 16219416 TI - Ganglioglioma with anaplastic recurrence of the neuronal element following radiotherapy. AB - Gangliogliomas are rare tumours in adults representing between 0.5% and 1% of all neuroepithelial tumours. They are of mixed cellularity comprised both of a neuronal and a glial population. Although malignant transformation is rare, and this mostly occurs in the glial element, we present a case of malignant neuroblastomatous transformation of a ganglioglioma. This is only the second ever case to our knowledge in the literature reporting the participation of the neuronal element in the malignant transformation of a ganglioglioma. In contrast with the previous published case, our patient has received post-operative therapeutic radiotherapy. The related literature is reviewed and the oncogenic role of therapeutic irradiation is discussed. PMID- 16219417 TI - Emission of volatile organic compounds from composting of different solid wastes: abatement by biofiltration. AB - Emission of volatile organic compounds (VOCs) produced during composting of different organic wastes (source-selected organic fraction of municipal solid wastes (OFMSW), raw sludge (RS) and anaerobically digested wastewater sludge (ADS) and animal by-products (AP)) and its subsequent biofiltration have been studied. Composting was performed in a laboratory scale composting plant (30l) and the exhaust gases generated were treated by means of a compost biofilter. VOCs concentration in the composting exhaust gases for each composting process ranged from 50 to 695 mg C m-3 for OFMSW (5:1), from 13 to 190 mg C m-3 for OFMSW (1:1), from 200 to 965 mg C m-3 for RS, from 43 to 2900 mg C m-3 for ADS and from 50 to 465 mg C m-3 for AP. VOCs emissions were higher during the beginning of the composting process and were not generally related to the biological activity of the process. These emissions corresponded to an average loading rate applied to the biofilter from 2.56 to 29.7 g C m-3 biofilter h-1. VOCs concentration in the exhaust gas from the biofilter ranged from 55 to 295 mg C m-3 for OFMSW (5:1), from 12 to 145 mg C m-3 for OFMSW (1:1), from 55 to 270 mg C m-3 for RS, from 42 to 855 mg C m-3 for ADS and from 55 to 315 mg C m-3 for AP. Removal efficiencies up to 97% were achieved although they were highly dependent of the composted waste. An important observation was that the compost biofilter emitted VOCs with an estimated concentration of 50 mg C m-3. PMID- 16219418 TI - Thermodynamic studies of the interaction at the solid/liquid interface between metal ions and cellulose modified with ethylenediamine. AB - The chelate molecule, ethylenediamine, has been incorporated onto the surface of cellulose by sequential reaction of cellulose fibres with phosphorous oxychloride followed by the chelating agent. The modified material (CelNN) retained its fibrous nature and was shown to be efficient at adsorbing divalent metal cations from water by complexation. Adsorption isotherms were determined for suspensions of CelNN in metal ion solutions of different concentrations, and the data were adjusted to fit the modified Langmuir equation. The maximum numbers of moles of Cu2+, Ni2+ and Zn2+ adsorbed per gram of modified cellulose were 1.64 x 10(-3), 5.25 x 10(-4) and 1.06 x 10(-3), respectively. The thermodynamic effects related to the adsorption of metal ion onto the cellulose surface were determined by calorimetric titration. Gibbs free energy was spontaneous for all interactions. The adsorption processes all exhibited endothermic enthalpy values and were accompanied by increases in entropy. PMID- 16219419 TI - Near and far field contamination modeling in a large scale enclosure: Fire Dynamics Simulator comparisons with measured observations. AB - The occurrence of a fire, no matter how small, often exposes objects to significant levels of contamination from the products of combustion. The production and dispersal of these contaminants has been an issue of relevance in the field of fire science for many years, though little work has been done to examine the contamination levels accumulated within an enclosure some time after an incident. This phenomenon is of great importance when considering the consequences associated with even low level contamination of sensitive materials, such as food, pharmaceuticals, clothing, electrical equipment, etc. Not only does such exposure present a localized hazard, but also the shipment of contaminated goods places distant recipients at risk. It is the intent of this paper to use a well-founded computational fluid dynamic (CFD) program, the Fire Dynamics Simulator (FDS), a large eddy simulation (LES) code developed by National Institute of Standards and Technology (NIST), to model smoke dispersion in order to assess the subject of air contamination and post fire surface contamination in a warehouse facility. Measured results are then compared with the results from the FDS model. Two components are examined: the production rate of contaminates and the trajectory of contaminates caused by the forced ventilation conditions. Each plays an important role in determining the extent to which the products of combustion are dispersed and the levels to which products are exposed to the contaminants throughout the enclosure. The model results indicate a good first order approximation to the measured surface contamination levels. The proper application of the FDS model can provide a cost and time efficient means of evaluating contamination levels within a defined volume. PMID- 16219420 TI - Fixed bed column study for Cd(II) removal from wastewater using treated rice husk. AB - A fixed bed of sodium carbonate treated rice husk was used for the removal of Cd(II) from water environment. The material as adopted was found to be an efficient media for the removal of Cd(II) in continuous mode using fixed bed column. The column having a diameter of 2 cm, with different bed depths such as 10, 20 and 30 cm could treat 2.96, 5.70 and 8.55 l of Cd(II) bearing wastewater with Cd(II) concentration 10 mg/l and flow rate 9.5 ml/min. Different column design parameters like depth of exchange zone, adsorption rate, adsorption capacity, etc. was calculated. Effect of flow rate and initial concentration was studied. Theoretical breakthrough curve was drawn from the batch isotherm data and it was compared with experimental breakthrough curve. An amount of 0.01 mol/l HCl solution was used for desorption of adsorption column. Column regeneration and reuse studies were conducted for two cycles of adsorption-desorption. PMID- 16219421 TI - Subtypes of neuronal nicotinic acetylcholine receptors involved in nicotine induced phosphorylation of extracellular signal-regulated protein kinase in PC12h cells. AB - Although many kinds of nicotinic acetylcholine receptor (nAChR) subtypes have been reported in the neuronal tissues, subtype differences in the nAChR-mediated intracellular signaling remains obscure. Using nAChR agonists and antagonists, the involvement of nAChRs in extracellular signal-regulated protein kinase (ERK) phosphorylation in PC12h cells was investigated. Cytisine and nicotine induced the phosphorylation of ERKs in a dose-dependent manner, whereas RJR-2403 had no effect. Cytisine, but not RJR-2403, also induced phosphorylation of CREB. Mecamylamine, dextromethorphan and 18-methoxycoronaridine inhibited nicotine induced ERK phosphorylation with much higher affinity than dihydro-beta erythroidine and alpha-conotoxin MII. These results suggest the involvement of alpha3beta4 nAChRs in ERK phosphorylation in PC12h cells. PMID- 16219422 TI - The majority of brain mast cells in B10.PL mice is present in the hippocampal formation. AB - In the healthy mammalian CNS, mast cells (MCs) are thought to be located mostly in the thalamus. In this study, we have systematically assessed the presence of MCs in the hippocampal formation (HF) and in the thalamus of normal male and female B10.PL mice. Giemsa(+) and Toluidine Blue(+) MCs were detected by histomorphometric analyses at perivascular and intraparenchymal sites of both the hippocampus and the entorhinal cortex. We found a mean number of 4.4 MCs in the HF of female and 3.3 MCs in male B10.PL mice. In contrast to the HF, no MCs were present in the thalamus of these mice. Notably, all HF-MCs showed immunoreactivity for Kit, the receptor for the MC growth and maturation factor SCF, as assessed by FITC-avidin/Kit double labelling. We demonstrate that the majority of brain MCs is found in the hippocampus and entorhinal cortex of B10.PL mice, though the total number of MCs is small compared to other mouse strains or rats. The presence of most brain MCs in the HF of B10.PL mice suggests a potential role of MCs in hippocampal physiology and pathology. PMID- 16219423 TI - An fMRI study of Chinese character reading and picture naming by native Korean speakers. AB - Chinese characters appear in the currently used Korean language, and the system used for writing system the Korean language consists of a mixture of the Korean alphabet and Chinese characters. In the present study, neural mechanisms involved in reading a single Chinese character words and naming pictures by Korean native speakers were investigated using a functional magnetic resonance imaging technique. The findings show a right hemispheric dominance within the occipito temporal and the left middle/medial frontal area for both reading Chinese characters and naming pictures. This should reflect the specific visual processing of reading Chinese characters. Additional activations in inferior frontal and cingulage gyrus were also observed. The activations of inferior parietal region and thalamus are of interest, since we assume that these activations are strongly related to the phonological status of single Chinese character words rather than two character words that are typically used by Korean native speakers. PMID- 16219424 TI - Inhibition of the RhoA/Rho kinase system attenuates catecholamine biosynthesis in PC 12 rat pheochromocytoma cells. AB - The small GTPase, RhoA, and its downstream effecter Rho-kinase (ROK) are reported to be involved in various cellular functions, such as myosin light chain phosphorylation during smooth muscle contraction and exocytosis. Indeed, growing evidence suggests that the RhoA/Rho-kinase pathway plays an important role in regulating exocytosis in these cells. However, it is not known whether the RhoA/Rho-kinase pathway has an effect on catecholamine synthesis. Using the rat pheochromocytoma cell line, PC12, we examined the effects of either Rho-kinase inhibitor (Y27632) or RhoA inhibitor (C3 toxin) on nicotine-induced catecholamine biosynthesis. We show that nicotine (10 microM) induces a significant, though transient, increase in RhoA activation in these cells. Treatment with either Y27632 (1 microM) or C3 toxin (10 microg/ml) significantly inhibited the nicotine induced increase of tyrosine hydroxylase (TH) mRNA and the corresponding enzyme activity. TH catalyzes the rate-limiting step in the biosynthesis of catecholamine. Y27632 significantly inhibited nicotine-induced phosphorylation of TH at Ser40 as well as Ser19, which are known to be phosphorylated by Ca(2+)/calmodulin kinase II. Furthermore, Y27632 (10 microM) as well as C3 toxin (10 microg/ml) significantly inhibited the nicotine-induced increase of TH at the protein level. Thus, we propose that activation of RhoA, and its downstream effecter Rho-kinase, is a prerequisite for catecholamine biosynthesis in PC12 cells. At the concentrations used in our experiments, Y27632 does not affect cAMP/PKA activity or PKC activity, indicating that the inhibitory effect of Y27632 can be attributed to the inhibition of Rho-kinase activity as observed in chromaffin cells. In contrast, neither Y27632 (10 microM) nor C3 toxin (10 microg/ml) significantly altered catecholamine secretion in PC12 cells. In conclusion, we have demonstrated that inhibition of the Rho/Rho-kinase pathway in chromaffin cells lowers TH activity, probably through CaMKII inhibition. By contrast, neither Y27632 nor C3 toxin affect the secretion of catecholamine. PMID- 16219426 TI - Mutant KRAS in aberrant crypt foci (ACF): initiation of colorectal cancer? AB - Since aberrant crypt foci (ACF) were first described in 1987, they have been the subjects of hundreds of papers; however, the debate continues about their role in colorectal tumorigenesis. This review focuses on the many phenotypic, genetic and epigenetic alterations in ACF that support the hypothesis that ACF are putative precursors of colorectal cancer in both humans and experimental animals. Human ACF, both with and without dysplasia, are monoclonal and display evidence of chromosomal instability. Both of these characteristics are shared by colorectal cancers. While most ACF do not have APC mutations, a large proportion has KRAS mutations and methylated SFRP1 and SFRP2 genes. This epigenetic inactivation gives rise to constitutive Wnt signaling in these putative precursors of colorectal cancer. PMID- 16219425 TI - The flexible evolutionary anchorage-dependent Pardee's restriction point of mammalian cells: how its deregulation may lead to cancer. AB - Living cells oscillate between the two states of quiescence and division that stand poles apart in terms of energy requirements, macromolecular composition and structural organization and in which they fulfill dichotomous activities. Division is a highly dynamic and energy-consuming process that needs be carefully orchestrated to ensure the faithful transmission of the mother genotype to daughter cells. Quiescence is a low-energy state in which a cell may still have to struggle hard to maintain its homeostasis in the face of adversity while waiting sometimes for long periods before finding a propitious niche to reproduce. Thus, the perpetuation of single cells rests upon their ability to elaborate robust quiescent and dividing states. This led yeast and mammalian cells to evolve rigorous Start [L.H. Hartwell, J. Culotti, J. Pringle, B.J. Reid, Genetic control of the cell division cycle in yeast, Science 183 (1974) 46-51] and restriction (R) points [A.B. Pardee, A restriction point for control of normal animal cell proliferation, Proc. Natl. Acad. Sci. U. S. A. 71 (1974) 1286 1290], respectively, that reduce deadly interferences between the two states by enforcing their temporal insulation though still enabling a rapid transition from one to the other upon an unpredictable change in their environment. The constitutive cells of multi-celled organisms are extremely sensitive in addition to the nature of their adhering support that fluctuates depending on developmental stage and tissue specificity. Metazoan evolution has entailed, therefore, the need for exceedingly flexible anchorage-dependent R points empowered to assist cells in switching between quiescence and division at various times, places and conditions in the same organism. Programmed cell death may have evolved concurrently in specific contexts unfit for the operation of a stringent R point that increase the risk of deadly interferences between the two states (as it happens notably during development). But, because of their innate flexibility, anchorage-dependent R points have also the ability to readily adjust to a changing structural context so as to give mutated cells a chance to reproduce, thereby encouraging tumor genesis. The Rb and p53 proteins, which are regulated by the two products of the Ink4a-Arf locus [C.J. Sherr, The INK4a/ARF network in tumor suppression, Nat. Rev., Mol. Cell Biol. 2 (2001) 731-737], govern separable though interconnected pathways that cooperate to restrain cyclin D- and cyclin E dependent kinases from precipitating untimely R point transit. The expression levels of the Ink4a and Arf proteins are especially sensitive to changes in cellular shape and adhesion that entirely remodel at the time when cells shift between quiescence and division. The Arf proteins further display an extremely high translational sensitivity and can activate the p53 pathway to delay R point transit, but, only when released from the nucleolus, 'an organelle formed by the act of building a ribosome' [T. Melese, Z. Xue, The nucleolus: an organelle formed by the act of building a ribosome, Curr. Opin. Cell Biol. 7 (1995) 319 324]. In this way, the Ink4a/Rb and Arf/p53 pathways emerge as key regulators of anchorage-dependent R point transit in mammalian cells and their deregulation is, indeed, a rule in human cancers. Thus, by selecting the nucleolus to mitigate cell cycle control by the Arf proteins, mammalian cells succeeded in forging a highly flexible R point enabling them to match cell division with a growth rate imposed by factors controlling nucleolar assembling, such as nutrients and adhesion. It is noteworthy that nutrient control of critical size at Start in budding yeast has been shown recently to be governed by a nucleolar protein interaction network [P. Jorgensen, J.L. Nishikawa, B.-J. Breitkreutz, M. Tyers, Systematic identification of pathways that couple cell growth and division in yeast, Science 297 (2002) 395-400]. PMID- 16219427 TI - Low incidence of mamographically detected microcalcification in breast cancer patients with lobular histology may be attributed to low levels of osteopontin. PMID- 16219428 TI - Factors predicting retention in treatment: 10-year experience of a methadone maintenance treatment (MMT) clinic in Israel. AB - The aims were to identify predictors of treatment retention in an Israeli methadone maintenance treatment (MMT) clinic, and to compare the findings to other international settings. We prospectively studied 492 patients admitted since 1993 through 10 years to an Israeli MMT clinic associated with a university affiliated tertiary care medical center. Analyses (Kaplan Meier and Cox regression) included methadone dose and urinalysis results (for methadone, cocaine, opiates, benzodiazepines, THC, amphetamines) of each patient in the first month and after 1 year in treatment (or during the last month if the stay was >3 months and <1 year) and patients' characteristics (modified ASI). The 1 year retention rate was 74.4%; 65.8% stopped opiate abuse after 1 year in treatment. On admission, 13.6% of patients had used cocaine: there was a net decrease of 61.6% after 1 year. Factors predicting prolonged retention in MMT treatment (Cox regression) were daily methadone dose of 100mg or greater, negative urine for opiates after 1 year, and being a parent on admission. We conclude that our good outcome results (high rate of retention after 1 year (74.4%), high proportion of opiate abuse cessation (65.8%), and net reduction in cocaine abuse, similar to normal standards in other MMT clinics elsewhere in the world, justify the expansion of the MMT clinic network in Israel in order to make treatment available to all those who need it. A protocol favoring higher methadone dosage as appropriate is recommended. PMID- 16219429 TI - Response to methadone maintenance treatment of opiate dependent patients with and without significant pain. AB - BACKGROUND: Both clinicians and researchers have expressed doubt that opiate dependent patients with significant pain can be effectively treated in methadone maintenance treatment (MMT) programs; however, little research exists on this topic. Patients who report significant pain in the month preceding entry to MMT present with a distinct and more severe pattern of polysubstance use, medical and psychosocial problems than do those without pain. The present study investigated the 1-year treatment outcomes of MMT patients with opiate dependence and pain. METHODS: Analyses were based on a national sample of 200 patients presenting in MMT programs for treatment of opiate dependence. Substance use and related problems were measured at treatment entry and 12 months later. Patients reported pain severity over the month preceding treatment entry. RESULTS: Compared to patients without significant pain, patients who reported significant pain at baseline (n = 103) showed similar substance-related functioning, but poorer psychosocial functioning at 1 year. CONCLUSIONS: Patients with and without significant pain experience comparable reductions in substance use when provided with standard care in MMT programs. However, additional medical and/or mental health treatment is needed for their pain and other problems. PMID- 16219430 TI - Molecular cloning and tissue-specific transcriptional regulation of the first peroxidase family member, Udp1, in stinging nettle (Urtica dioica). AB - A full-length cDNA clone, designated Udp1, was isolated from Urtica dioica (stinging nettle), using a polymerase chain reaction based strategy. The putative Udp1 protein is characterized by a cleavable N-terminal signal sequence, likely responsible for the rough endoplasmic reticulum entry and a 310 amino acids mature protein, containing all the important residues, which are evolutionary conserved among different members of the plant peroxidase family. A unique structural feature of the Udp1 peroxidase is defined into the short carboxyl terminal extension, which could be associated with the vacuolar targeting process. Udp1 peroxidase is differentially regulated at the transcriptional level and is specifically expressed in the roots. Interestingly, wounding and ultraviolet radiation stress cause an ectopic induction of the Udp1 gene expression in the aerial parts of the plant. A genomic DNA fragment encoding the Udp1 peroxidase was also cloned and fully sequenced, revealing a structural organization of three exons and two introns. The phylogenetic relationships of the Udp1 protein to the Arabidopsis thaliana peroxidase family members were also examined and, in combination with the homology modelling approach, dictated the presence of distinct structural elements, which could be specifically involved in the determination of substrate recognition and subcellular localization of the Udp1 peroxidase. PMID- 16219431 TI - Species-specific variation of alternative splicing and transcriptional initiation in six eukaryotes. AB - The genome-wide detection of alternative splicing and transcriptional initiation (ASTI) was conducted in six eukaryotes (human, mouse, fruit fly, nematode, cress and rice) whose genome sequencing has been completed or nearly completed. Transcriptional isoforms were collected by mapping a batch of full-length cDNA sequences onto the respective cognate genomic sequences. Isoforms mapped on the same gene locus were compared pair-wise, ASTI patterns were segmented into minimal spans, and then the minimal patterns (ASTI units) were classified into unique types, such as the cassette type or the alternative donor site. All these procedures were performed automatically under the same conditions so that the results obtained from different species could be compared directly. The fraction of loci that underwent ASTI of the total mapped loci was the largest for mammals and fruit fly, and the smallest for plants. Exactly the same trend was observed for the number of unique ASTI types found in each species. The observed fractional representations of the ASTI types were similar between evolutionarily close species, such as human and mouse or cress and rice. On the other hand, the relative orders of abundance in individual ASTI type were considerably different between evolutionarily distant species, such as between mammals and plants. In human and mouse, alternative splicing other than the retained introns tended to occur within the protein coding sequence (CDS) regions rather than within the untranslated regions (UTRs), whereas this tendency was obscure in the other four species. In all the species examined, the difference in alternative exon lengths was most likely in multiples of three, and this tendency was most prominent when the alternative exons were embedded within the CDSs. These observations are generally consistent with the idea that higher organisms utilize the ASTI mechanisms more extensively and in a more complicated manner than lower organisms, and that ASTI actively participates in the enhancement of the functional and structural diversity of products generated from a limited number of genes on a genome. PMID- 16219432 TI - Cellular uptake and activation characteristics of naked plasmid DNA and its cationic liposome complex in human macrophages. AB - Plasmid DNA (pDNA) is an important macromolecular therapeutic agent suitable for DNA-based therapies, such as non-viral gene therapy and DNA vaccination. Unmethylated CpG motifs abundant in bacterial DNA, but not in vertebrate DNA, are known to trigger an inflammatory response, which inhibits transgene expression, while modulating immunological consequences following vaccination. We studied cellular uptake and activation characteristics of naked pDNA and its cationic liposome complex in human macrophage-like cells. The present study has demonstrated that naked pDNA was recognized by human macrophage-like cells via specific mechanisms for polyanions. Moreover, it has shown that pDNA complexed with cationic liposomes activates human macrophage-like cells to induce the production of tumor necrosis factor-alpha (TNF-alpha) in a CpG motif-independent manner, while any types of naked DNA could not induce TNF-alpha production from these cells, regardless of the presence of CpG motifs in pDNA or oligonucleotide (ODN). These findings form an important basis for DNA-based therapies including gene therapy and DNA vaccination. PMID- 16219433 TI - The novel formulation design of O/W microemulsion for improving the gastrointestinal absorption of poorly water soluble compounds. AB - The design of the novel O/W microemulsion formulation, which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds was examined. Using medium chain fatty acid triglyceride (MCT), diglyceryl monooleate (DGMO-C), polyoxyethylene hydrogenated castor oil 40 (HCO-40), ethanol and PBS (pH 6.8) as an oil phase, a lipophilic surfactant, a hydrophilic surfactant, a solubilizer and an aqueous phase, at the mixture ratio of 5%/1%/9%/5%/80% (w/w), respectively, the O/W microemulsion with an average particle diameter of 20 nm or less was prepared. Moreover, for nine kinds of poorly water soluble compounds, such as Ibuprofen, Ketoprofen, Tamoxifen, Testosterone, Tolbutamide and other new compounds, the solubility to water was increased from 60 to 20,000 times by this O/W microemulsion formulation. The AUCs in plasma concentration of Ibuprofen and a new compound, ER-1039, following single oral administration of these compounds as the O/W microemulsion to fasted rats were equivalent to that of solution administration or increased by nine and two times that of suspension administration, respectively. Accordingly, this novel O/W microemulsion is a useful formulation, which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds. PMID- 16219434 TI - Evaluation of the taste and smell of bottled nutritive drinks. AB - The purpose of this study was to evaluate the palatability of 15 bottled nutritive drinks, all commercially available in the Japanese market, using data from artificial taste and odor sensors. In gustatory sensation tests, well trained healthy volunteers were asked to score the drinks in terms of palatability and of the four basic tastes. The results suggest that overall palatability is positively correlated with sourness intensity and fruitiness (R=0.82 and 0.86, respectively) and negatively correlated with bitterness intensity and the tasting of medicinal plants (R=-0.85 and -0.80, respectively). The sourness and bitterness intensity could be predicted by taste sensor and fruitiness could be predicted by odor sensor, respectively. By performing principal component analysis of the taste sensor data, the 15 drinks could be classified into four groups. The group classified as being predominantly sour had the highest palatability score, 3.8. By principal component analysis of odor sensor data, the drinks could also be classified into four groups and this time the group with a fruity flavor (smell) showed the highest palatability score, 3.4. In the combined analysis of both taste and odor data, products containing medicinal plants showed the lowest palatability. Finally, the combined usage of the taste and odor sensors gave rise to a three-group classification. Thus, not only the taste sensor but also the odor sensor may be useful in evaluating the palatability of bottled nutritive drinks. PMID- 16219435 TI - Menthol facilitates the skin analgesic effect of tetracaine gel. AB - The aim of this study is to observe the effect of menthol on the percutaneous penetration and skin analgesic action of tetracaine gel (T-gel). Anesthetic gels containing 4% tetracaine in carbomer vehicle with and without menthol were prepared. The menthol penetration-enhanced gel conferred significantly higher diffusion of tetracaine across full-thickness mouse skin than non-penetration enhanced gel, in a dose-dependent manner. The inter-cellular spaces of the stratum corneum in skin treated with menthol penetration-enhanced gel became extended as compared with those in non-penetration-enhanced gel. This may suggest that menthol's action was related to the changes of the epidermis ultra structures. An enlarged inter-cellular space, per se, would allow a better passage to tetracaine. To determine the efficacy of menthol penetration-enhanced tetracaine gel in the management of pain, a double-blind, placebo-controlled, randomized controlled trial (RCT) design was used. The mean verbal pain scores (VPS) were significantly lower in volunteers treated with penetration-enhanced tetracaine gel than those in volunteers receiving non-penetration-enhanced tetracaine gel or placebo. Menthol improved the analgesic efficacy of the tetracaine 4% gel in part through enhanced percutaneous permeation. PMID- 16219436 TI - Extremely rapid induction of outer hair cell apoptosis in the chinchilla cochlea following exposure to impulse noise. AB - We have reported the presence of OHC apoptosis and necrosis in the organ of Corti following exposure to intense noise. The current study was designed to investigate the rapidity and the initial pattern of outer hair cell (OHC) death induced by exposure to impulse noise. Chinchillas were exposed to 75 pairs of impulse noise at 155 dB peak sound pressure level presented over a time period of 75 s. At 5 or 30 min after the noise exposure, the cochleae were examined for morphological and biological indicators of apoptosis and necrosis. In the cochleae collected within 5 min after the 75-s noise exposure, there were clear signs of nuclear condensation and cell body shrinkage, suggesting the presence of OHC apoptosis. Apoptotic OHCs were further detected by positive staining of TUNEL and caspase-3 assays. In contrast to the rapid development of nuclear condensation, appearance of nuclear swelling, a necrotic phenotype, appeared at 30 min after the noise exposure. The results of the study demonstrate that induction of OHC apoptosis after the noise exposure is an extremely rapid process. PMID- 16219437 TI - Coeloglossum viride var. bracteatum extract attenuates D-galactose and NaNO2 induced memory impairment in mice. AB - The aim is to study the neuroprotective effects of CE, a special extract from Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.), on memory deficits in senescent mice induced by consecutive injection of d-galactose and NaNO(2) for 60 days. Oral administration of CE (2.5 mg kg(-1), 5 mg kg(-1)) started from 47th day of the experiment. Water maze test was used to evaluate the learning and memory function of mice. The contents of MDA and GSH and the activities of SOD and GSH-px in brain tissue were determined using different biochemical methods. The alterations in hippocampus morphology were assessed. Immunoreactive cells of Tau-2 and NT-3 in the hippocampus were counted by immunohistochemical staining. The results indicated that combined injection of d-galactose and NaNO(2) induced memory impairment of mice. In addition, SOD and GSH-px activities and GSH level decreased while MDA level increased in mouse brain. Tau-2 positive neurons increased while NT-3 positive neurons decreased remarkably in the hippocampus. Administration of CE for 14 days significantly improved the cognitive deficits and the biochemical markers mentioned above and reduced the histological lesions in mouse brain. These results suggest that CE is worth testing for further preclinical study aimed for senescence and dementia. PMID- 16219438 TI - Protective effect of Polygonum multiflorum Thunb on amyloid beta-peptide 25-35 induced cognitive deficits in mice. AB - Amyloid beta protein (Abeta) may be neurotoxic during the progression of Alzheimer's disease by eliciting oxidative stress. This study was designed to determine the effect of Polygonum multiflorum Thunb water extract (PWE) on Abeta25-35-induced cognitive deficits and oxidative stress in mice. Mice were fed experimental diets comprising either 0.5 or 1% PWE for 4 weeks, and then received a single intracerebroventricular (i.c.v.) injection of Abeta25-35 (10 microg/mouse). Behavioral changes in the mice were evaluated using passive avoidance and water-maze tests. The consumption of PWE significantly ameliorated the cognitive deficits caused by i.c.v. injection of Abeta25-35. The Abeta25-35 treatment accelerated the lipid peroxidation, and PWE attenuated the Abeta induced increase in brain levels of thiobarbituric acid reactive substances. There was an increase in glutathione peroxidase activity in PWE-treated groups. The acetylcholinesterase activity in the brain and serum was lower in PWE supplemented groups than in the only Abeta-injected group. These findings suggest that PWE exerts a preventive effect against cognitive deficits induced by Abeta25 35 accumulation in Alzheimer's disease, and that this effect is mediated by the antioxidant properties of PWE. PMID- 16219439 TI - Anti-inflammatory and anti-ulcerogenic properties of Stachytarpheta cayennensis (L.C. Rich) Vahl. AB - In the present work, the anti-inflammatory and gastroprotective properties of ethanolic extracts of Stachytarpheta cayennesis (L.C. Rich) Vahl (Verbenaceae) were assessed. Chromatographic analysis of the crude ethanolic extract, SC01, revealed high concentrations of the iridoid ipolamiide, whereas the SC02, the second ethanolic extract, presented the arylpropanoid verbacoside as a major constituent. The oral administration of SC01 (100 mg/kg) into Swiss mice failed to inhibit paw oedema and pleural exudation induced by carrageenan and zymosan, whereas SC02 (100 mg/kg, p.o.) inhibited oedema and protein extravasation in all instances. Both extracts inhibited total leukocyte accumulation into the pleural cavity 4 and 24h after the intrathoracic (i.t.) injection of carrageenan, due to the inhibition of neutrophil and mononuclear cell influx, whereas only SC02 was able to inhibit leukocyte mobilization induced by zymosan (100 microg/cavity, i.t.). SC02 inhibited LPS (250 ng/cavity)-induced total leukocyte, neutrophil and eosinophil accumulation in the pleural cavity, whereas SC01 selectively inhibited neutrophil influx. In addition, our data indicates that the extract SC02 presents an important anti-ulcerogenic activity, since it inhibited diclofenac-induced (100 mg/kg, p.o.) gastric ulcera. Overall, these data provide evidence for the anti-inflammatory and gastroprotective properties of Stachytarpheta cayennensis, supporting its use in folk medicine for such purposes. PMID- 16219440 TI - Argentine folk medicine: genotoxic effects of Chenopodiaceae family. AB - Chenopodium ambrosioides L. and Chenopodium multifidum L. (Chenopodiaceae), common name: Paico, are medicinal plants. They are aromatic shrubs growing in South America. For centuries, they have been used due to its medicinal properties. However, there are few reports in literature about the genotoxic effects of these plants. There for, the aim of these work is the evaluation of genetic damage induced by decoction and infusion of this plants which were assayed in different concentrations (1, 10, 100, 1,000 microL extract/mL culture), by addition of the extract to human lymphocyte cell cultures, negative controls were included. The endpoints evaluated were chromosomal aberrations (CA), sister chromatid exchanges (SCE), cell proliferation kinetics (CPK) and mitotic index (MI). The repeated measure analysis of variance was used for statistic evaluation of the results. The results showed: (a) statistical increase in the percentage of cells with CA and in the frequency of SCE when cultures were exposed to both aromatic plants, (b) a decrease in MI of both Paicos assayed, although no modification in the CPK values was observed, (c) no effect was noticed in the analysis of Chenopodium album L., which was used as negative control of the essential oil. These results suggest a cyto and genotoxic effect of Chenopodium ambrosioides and Chenopodium multifidum aqueous extracts related to the essential oil of the plant (as Chenopodium album did not perform). PMID- 16219441 TI - Studies on the formation, temporal evolution and forensic applications of camera "fingerprints". AB - A series of experiments was conducted by exposing negative film in brand new cameras of different make and model. The exposures were repeated at regular time intervals spread over a period of 2 years. The processed film negatives were studied under a stereomicroscope (10-40x) in transmitted illumination for the presence of the characterizing features on their four frame-edges. These features were then related to those present on the masking frame of the cameras by examining the latter in reflected light stereomicroscopy (10-40x). The purpose of the study was to determine the origin and permanence of the frame-edge-marks, and also the processes by which the marks may probably alter with time. The investigations have arrived at the following conclusions: (i) the edge-marks have originated principally from the imperfections received on the film mask from the manufacturing and also occasionally from the accumulated dirt, dust and fiber on the film mask over an extended time period. (ii) The edge profiles of the cameras have remained fixed over a considerable period of time so as to be of a valuable identification medium. (iii) The marks are found to be varying in nature even with those cameras manufactured at similar time. (iv) The influence of f/number and object distance has great effect in the recording of the frame-edge marks during exposure of the film. The above findings would serve as a useful addition to the technique of camera edge-mark comparisons. PMID- 16219442 TI - Activity and tissue-specific expression of lipases and tumor-necrosis factor alpha in lean and obese cats. AB - Post-heparin plasma activity of lipoprotein lipase (LPL) and hepatic lipase (HL), and fat and muscle activity of LPL were measured in neutered lean and obese cats. Lipoprotein lipase, hormone-sensitive lipase (HSL), and tumor necrosis factor a (TNF) mRNA were measured in muscle and fat tissue with real-time PCR using primers for feline LPL, HSL, and TNF. Lipoprotein lipase plasma and fat activity and fat mRNA levels were significantly lower (50, 80, and 50%, respectively) in obese cats than lean cats, whereas the muscle/fat ratio of LPL was significantly higher in obese compared to lean cats. The activity of HL was not different between the groups. Hormone-sensitive lipase mRNA levels were significantly higher in obese than lean cats. The level of fat TNF also was significantly higher in obese cats than in lean cats, whereas the level in muscle was not different. The lower LPL activity and mRNA expression in fat and the higher LPL and HSL mRNA expression in muscle in obese cats compared to lean cats expectedly favor a redistribution of fatty acids from fat to muscle tissue where they can be deposited or used for energy in times of need. Tumor necrosis factor alpha may regulate this repartitioning process through suppression of adipocyte LPL. PMID- 16219443 TI - Hormonal and metabolic adaptation to fasting: effects on the hypothalamic pituitary-ovarian axis and reproductive performance of rabbit does. AB - To assess the impact of acute caloric shortage on reproduction, rabbit does were either fed ad libitum (control, AL), or fasted for 24 (STF) or 48 h (LTF) before induction of ovulation with GnRH injection. Blood samples were collected during the last 3 h of fasting, and the following 4 h after GnRH injection, when feed was provided again, to measure plasma concentrations of LH, estradiol-17beta, leptin, insulin, T3, corticosterone, glucose, and NEFA. Before re-feeding, plasma leptin, insulin, and T3 concentrations were lower (P < or = 0.01) in both fasted groups than in controls, but then gradually increased following realimentation to match those of controls. During fasting, corticosterone levels were higher (P < or = 0.01) in LTF than in STF and AL does, but decreased to control values soon after realimentation. During fasting, plasma glucose concentrations did not differ among groups, but upon re-feeding they markedly increased (P < or= 0.01) both in STF and LTF does. NEFA levels were also more elevated (P < or = 0.01) in fasted rabbits than in controls, and rapidly decreased (P < or = 0.01) after re feeding. Following GnRH injection, LH peak was lower (P < or = 0.01) in LTF than in AL and STF does. Estradiol-17beta showed higher pulse frequency and amplitude in AL than in STF and LTF does. Compared to controls, receptivity rate of STF and LTF artificially inseminated does declined respectively by -20.5% (P < or = 0.05) and -22.7%, and fertility rate by -23.9% (P < or = 0.05) and 21.4%, but no difference was found in ovulation rate. In summary, nutritional status of does, as modified by fasting, greatly influenced fertility, metabolic and reproductive hormones. PMID- 16219444 TI - Pre-pubertal measurements of the somatotrophic axis as predictors of milk production in Holstein-Friesian dairy cows. AB - This study investigated possible relationships between measurements of the somatotrophic axis in pre-pubertal dairy calves and subsequent milk yields. Endogenous growth hormone (GH) release was measured through a fed and fasted period in fifty 6-month-old Holstein-Friesian heifers and they were then challenged with growth hormone-releasing factor (GRF) to assess their GH release pattern. Insulin-like growth factor-I (IGF-I), insulin and glucose concentrations were measured in relation to time of feeding. Cows were subsequently monitored through their first three lactations to record peak and 305-day milk yields. In the first lactation, milk energy output for the first 120 days of lactation was also calculated. The mean 305-day milk yield increased from 7417 +/- 191 kg in the first lactation (n=37) to 8749 +/- 252 kg in the third (n=25). There were no significant relationships between any measures of GH secretion and peak or 305 day yield in any lactation. A highly significant positive relationship was established between the GH peak measured 10 min post-GRF challenge and 120-day milk energy values in the first lactation. This relationship was, however, only present in the sub-population of 12 cows culled after one or two lactations and was absent in the 25 animals remaining for the third lactation. There were no significant relationships between pre-pubertal IGF-I and fed or fasted insulin or glucose concentrations and any subsequent measurement of yield. The usefulness of GH secretagogue challenges in calves as a predictive test for future milk production is thus limited but may have some bearing on nutrient partitioning and longevity. PMID- 16219445 TI - T-screen to quantify functional potentiating, antagonistic and thyroid hormone like activities of poly halogenated aromatic hydrocarbons (PHAHs). AB - The present study investigates chemical thyroid hormone disruption at the level of thyroid hormone receptor (TR) functioning. To this end the (ant)agonistic action of a series of xenobiotics was tested in the newly developed T-screen. This assay makes use of a GH3 rat pituitary cell line, that specifically proliferates when exposed to 3,3',5-triiodo-L-thyronine (T3). The growth stimulatory effect is mediated via T3-receptors. (Ant)agonistic and potentiating action of compounds was studied in absence and presence of T3 at its EC50 level (0.25 nM). The compounds tested included the specific TR-antagonist amiodarone, as well as a series of brominated diphenyl ethers (BDEs), including specifically synthesized BDEs with a structural resemblance to 3,5-diiodo-L-thyronine (T2), T3 and T4 (3,3',5,5'-tetraiodo-L-thyronine). The results obtained reveal that only BDE206 and amiodarone are specific antagonists. Interestingly some compounds which did not respond in the T-screen in absence of T3, potentiated effects when tested in combination with T3. This points at possibilities for disruption at the TR in vivo, where exposure generally occurs in presence of T3. Altogether the results of the present study show that the newly developed T-screen can be used as a valuable tool for identification and quantification of compounds active in disturbing thyroid hormone homeostasis at the level of TR-functioning. PMID- 16219446 TI - Assessment of the phototoxic hazard of some essential oils using modified 3T3 neutral red uptake assay. AB - When substances are developed in the aim to be a constituent of personal care products, and to be applied on the skin, it is necessary to carry out an assessment of potential phototoxic hazard. Phototoxicity is skin reaction caused by concurrent topical or systemic exposure to specific molecule and ultraviolet radiation. Most phototoxic compounds absorb energy particularly from UVA light leading to the generation of activated derivatives which can induce cellular damage. This type of adverse cutaneous response can be reproduced in vitro using different models of phototoxicity such as the validated 3T3 Neutral Red Uptake (NRU) phototoxicity assay. In the present study we utilised two different cell lines (the murine fibroblastic cell line 3T3 and the rabbit cornea derived cell line SIRC) to compare the photo-irritation potential of a strong phototoxic compound, chlorpromazine, to a weaker composite, such as 8-methoxypsoralen and Bergamot oil. After comparison of the different systems, five other essential oils were tested with both cell lines. Cellular damage was evaluated by the NRU cytotoxicity test or by MTT conversion test. PMID- 16219447 TI - Developmental changes in concentrations and distributions of neurotrophins in the monkey cerebellar cortex. AB - Neurotrophins are involved in the survival, differentiation, migration and neurite outgrowth of various neuronal populations. Neurotrophins and their receptors are widely expressed in the developing cerebellum of various experimental animals. To gain some insight into the possible roles played by these molecules in monkey cerebellum, we examined the protein levels of BDNF, NT 4/5 and NT-3 and distributions of those neurotrophins and TrkC, a high affinity receptor for NT-3, in the cerebellum of developing macaque monkeys using ELISAs and immunohistochemical methods. We found that the level of BDNF increased during development, while the level of NT-3 was higher during embryonic stages and decreased toward adulthood. The level of NT-4/5 increased from embryonic stages to infant stages and gradually declined with age. Among the three neurotrophins, BDNF immunoreactivity was found in all kinds of cerebellar neurons, including all inhibitory interneurons, throughout the postnatal periods examined, indicating that BDNF may be an essential factor for the maintenance of cerebellar neural functions. The Bergmann glial fibers and the internal part of the external granule cell layer were strongly NT-3 immunopositive at the early postnatal stages, and more weakly immunoreactive toward adulthood. In addition, we found that the premigratory precursors of the granule cells were TrkC immunopositive at early postnatal stages. These findings suggest that NT-3 in Bergmann glial fibers may be involved in the migration of the premigratory granule cells. PMID- 16219448 TI - Prediction of the pharmacokinetics of succinylated human serum albumin in man from in vivo disposition data in animals and in vitro liver slice incubations. AB - Suc-HSA is a potent HIV-inhibitor with possible application in man. To facilitate the assessment of dosing regimens for future phase I clinical studies, we predicted the pharmacokinetic properties of Suc-HSA in man. Slices prepared from rat, monkey and human liver were incubated with succinylated albumin, and the maximum uptake rate V(m) and Michaelis-Menten constant K(m) were calculated. The pharmacokinetics after multiple doses of Suc-HSA were studied in rats. The pharmacokinetic parameters of Suc-HSA in man were predicted from the results and data from literature, using pharmacokinetic modeling and interspecies scaling techniques, and potential intravenous dose regimens for HIV treatment in man were calculated. On the basis of in vitro uptake studies in rat, monkey and human liver slices and in vivo disposition data in monkey (data from earlier study) and rat, we predicted the following parameters for liver uptake in humans: V(m) 82.5 microg h(-1) kg(-1) and K(m) 0.228 microg ml(-1). The predicted steady-state concentration after daily intravenous bolus doses of 1 mg kg(-1) is between 4 and 30 microg ml(-1), i.e. well above the IC50 of about 0.4 microg ml(-1). Additional loading doses of 8 mg kg(-1) in total are needed to reach steady-state within a few days. PMID- 16219449 TI - A strategy for preclinical formulation development using GastroPlus as pharmacokinetic simulation tool and a statistical screening design applied to a dog study. AB - The aim of this paper is to propose a pharmaceutical risk assessment strategy that goes beyond the usual characterisation of a clinical candidate molecule according to the biopharmaceutical classification system (BCS). This strategy was evaluated for a new CNS drug with poor solubility and good permeability. In a first step, GastroPlus was used to simulate the absorption process based on preformulation data. These input data involved a physicochemical drug characterisation including drug solubility measurements in simulated physiological media, as well as permeability determination. Further computer simulations were conducted to determine the sensitivity to changes of selected input values. Thus, oral bioavailability prediction was studied as a function of the particle size and drug solubility. The second part of the presented strategy for preclinical formulation development was to test specially designed formulations in a 2(3) screening factorial plan using the dog as the animal model. The factors were the dosage form, food effect and dose strength. One of the two experimental formulations was a capsule filled with the micronised drug, whereas the other formulation was a surfactant solution of the drug. Accordingly, a "worst case" formulation was compared with a "best case" drug solution over the clinically relevant dose range in fasted and fed dogs. The results of the computer simulation indicated that a fraction of the dose is dissolved in the stomach and precipitates partially in the small intestine. The simulation predicted almost full drug absorption during the GI transit time. Interestingly, the simulation implies that stomach drug solubility had little impact on overall fraction absorbed. The results also showed that changes of particle size and reference solubility within two orders of magnitude hardly affected the oral bioavailability. This in silico deduction was subsequently compared with the results of the dog studies. Indeed a surfactant drug solution showed no clear biopharmaceutical superiority over a solid capsule formulation on the average of both dose strengths in fasted and fed dogs. Despite the substantial variability of the in vivo data, the factorial screening design indicated marginal significant interaction between the dose level and feeding status. This can be viewed as a flag for the planning of further studies, since a potential effect of one factor may depend on the level of the other. In summary, the GastroPlus simulation together with the statistically designed dog study provided a thorough biopharmaceutical assessment of the new CNS drug. Based on these findings, it was decided to develop a standard granulate in capsules for phase I studies. More sophisticated formulation options were abandoned and so the clinical formulation development was conducted in a cost-efficient way. PMID- 16219450 TI - [Neuropediatric approach to autism]. AB - Autism is defined by 3 main criteria: disturbance of reciprocal social interaction, disturbance of communication (including language comprehension and spoken language) and disturbance of normal variation in behaviour and imaginative activities; an onset before age 36 months is also required. The neuropediatric contribution to autism is dominated by the search for an underlying organic etiology, especially if there are arguments for an associated encephalopathy: ante- or perinatal medical history, dysmorphic signs, skin spots, neurological abnormalities, somatic abnormalities compatible with a neurometabolic disorder. The main associated conditions with autism are: chromosome anomalies, monogenic syndrome (including fragile X syndrome), neurocutaneous syndromes, epileptic encephalopathies, neurometabolic diseases, and dystrophinopathies. The identification of an associated medical condition to autism is primordial in prospect of genetic counselling, and by the change induced in familial perception of autism. PMID- 16219451 TI - [Drugs news]. PMID- 16219452 TI - [Hypernatremic dehydration in children: retrospective study of 105 cases]. AB - The hypernatremic dehydration defined by a serum sodium concentration> or = to 150 mmol/l, is a particular form of acute dehydration and constitutes a medical emergency requiring a prompt and adequate diagnosis and management. PURPOSE: To precise the epidemiological profile, course, causes and therapeutic particularities of hypernatremic dehydration in children. POPULATION AND METHODS: Retrospective review of 105 children admitted in the general Paediatrics department of the Fattouma Bourguiba university hospital in Monastir (Tunisia), for hypernatremic dehydration between January 1st 1990 and December 31 2002. RESULTS: Hypernatremic dehydration represented 11.51% of all kinds of dehydration. The mean age was 6.5 months with a small male predominance. The socio-economic level of the parents was good in 62.8% of cases. Half of the children were in shock. Severe dehydration was present in 87.6% of cases and neurological signs were observed in 77.14% of cases. The initial mean serum sodium concentration was 159 mmol/L. Acidosis and acute renal failure were associated respectively in 97.2% and 76.2% of cases. Prominent cause of hypernatremic dehydration was diarrhoea (94.3%). Intravenous rehydration with 5% glucose solution at the average of 147 ml/kg/day and containing a mean sodium level of 42 mmol/L was performed in 74% of cases. In most cases (84.1%) serum sodium was normalized within the first 72 hours. Complications were noted in 5.7% of cases and mortality rate was 11.4%. CONCLUSION: Hypernatremic dehydration was common in infant and the prominent cause is still dominated by diarrhoea in our country. The management of hypernatremic dehydration is based on oral or intravenous rehydration and plasma expanding fluids when shock is present or imminent. The serum sodium concentration should be gradually corrected and should not exceed 0.5 mmol/L/h. Prevention is based on the pursuit of breastfeeding and the use of oral rehydration solution in infantile diarrhoeas. PMID- 16219453 TI - Effect of a non-adjustable oral appliance on upper airway morphology in obstructive sleep apnoea. AB - BACKGROUND: To evaluate the effect of oral appliance (OA) on upper airway morphology and its relationship with treatment response in subjects with obstructive sleep apnoea (OSA). METHODS: Symptomatic OSA subjects were recruited. Non-adjustable OA was custom made. Variables examined at baseline and while wearing the device at 2 months included polysomnographic data, computed tomographic measurements of upper airway cross sectional area at level of velopharynx (VA) and hypopharynx (HA), upper airway volume, and cephalometric parameters. Treatment outcome was based on post-treatment apnoea-hypopnoea index (AHI). RESULTS: Forty patients were recruited and 23 (7 women) completed the study. They were middle-aged (49, 40-58 years) (median, interquartile range) and overweight (BMI 26, 23.3-29.5 kg/m2), with moderate OSA (AHI 26.4, 14.1-36). The overall post treatment AHI was 8.4 (2.4-12.5), with 14 (61%) patients showing good response (AHI<10), and the other 9 patients showing moderate response (>50% reduction in AHI but still 10). OA decreased the cross-sectional area of the HA (P=0.046), showed a trend of decreasing the ratio of cross-sectional area of the HA to cross-sectional area of the VA (P=0.053) and significantly increased the overall upper airway volume (P=0.006, n=11). No significant relationship between upper airway parameters and treatment outcome was identified. CONCLUSIONS: OA altered upper airway morphometry towards a profile consistent with decreased propensity to collapse, which may thus have contributed to improvement of OSA. PMID- 16219454 TI - Systemic effects of COPD. AB - Chronic obstructive pulmonary disease (COPD) is characterised by a range of pathological changes of the respiratory system, including airflow limitation secondary to structural changes of the small airways and loss of alveolar attachments, inflammation, ciliary dysfunction, and increased mucous production. COPD also has significant systemic consequences. The relationships between these pulmonary and nonpulmonary morbidities are not fully understood, and this further complicates the assessment of disease severity and prognosis. Although improving lung function and disease symptoms have been the main focus of COPD management, these parameters alone do not reflect the full burden of disease. More recent endeavours have highlighted the potential role of addressing physical limitations imposed by systemic alterations. It is evident that systemic manifestations are common in COPD. Indeed, many patients demonstrate a gradual and significant weight loss that exacerbates the course and prognosis of disease. This weight loss is often accompanied by peripheral muscle dysfunction and weakness, which markedly contribute to exercise limitation and impaired quality of life. Weight loss has been postulated to be the result of a high metabolic rate that is not compensated for by increased dietary intake. The cause of this elevated metabolism is a matter of much debate, and several factors have been implicated. Similarly, the processes underlying depletion of muscle mass and function have not been fully delineated. The impact of the systemic manifestations of COPD is substantial, and although many attempts have been made to elucidate the mechanisms underlying these manifestations, there are important questions, which remain to be answered. An increase in our understanding in this field will doubtless highlight potential therapeutic targets, and assist in guiding future therapeutic development. PMID- 16219455 TI - Association of the ASP299GLY TLR4 polymorphism with COPD. AB - Bacterial infection and colonization plays an important role in COPD. The inflammatory response to these bacteria is mediated by Toll-like receptors. The Asp299Gly polymorphism of the Toll-like receptor-4 (TLR4) has been shown to be associated with decreased lipopolysaccharide (LPS) signal transduction resulting in impaired antimicrobial defense. Because altered TLR4 signalling may facilitate bacterial infection, we clinically phenotyped and genotyped 152 patients with COPD (including 24 non-smokers), and 444 healthy controls for the presence of the Asp299Gly polymorphism. Frequencies of the TLR4 Gly allele (4% vs. 8% in controls, odds ratio (OR) 2.24 (95% confidence interval (95%CI) 1.17-4.3)) as well as TLR4 Gly genotype (6% vs. 13% in controls, OR 2.39 (95%CI 1.20-4.79)) were significantly decreased among the patients with COPD. The TLR4 Gly allele was not detected at all in a subgroup of non-smoking patients (n=24). We conclude that the frequency of the Asp299Gly polymorphism is decreased in COPD patients. Unaltered LPS signal transduction by TLR4 may be important for the development of COPD. PMID- 16219456 TI - The mitochondrial death squad: hardened killers or innocent bystanders? AB - Since the discovery that formation of the apoptosome in mammalian cells is triggered by cytochrome c released from the mitochondria, many other mitochondrial proteins have been suspected to be part of a conspiracy to cause cell death. AIF, EndoG, ANT, cyclophilin D, Bit1, p53AIP, GRIM-19, DAP3, Nur77/TR3/NGFB-1, HtrA2/Omi and Smac/Diablo have all been convicted as killers, but new genetic technology is raising questions about their guilt. Gene knockout experiments suggest that many were wrongly convicted on circumstantial evidence, and just happened to be in the wrong place at the wrong time. PMID- 16219457 TI - Modeling of spherical fluorescent glucose microsensor systems: design of enzymatic smart tattoos. AB - A two-substrate mathematical model of microspherical optical enzymatic glucose sensors is presented. The sensors are based on the well-known oxidation of glucose by glucose oxidase, and are constructed by the encapsulation of glucose oxidase within hydrogel microspheres coated with ultrathin polyelectrolyte multilayer films. In order to measure glucose via changes in oxygen concentration, a fluorescent oxygen indicator is co-encapsulated with the enzyme. The model was used to predict the temporal and spatial distributions of glucose and oxygen within the sphere for step increases in bulk glucose concentration. In addition, the model was used to observe the effect of varying sensor parameters, namely sphere size, film thickness, enzyme concentration, and mass transport of substrate and co-substrate within the sphere and film coatings, on the response of the sensors. A major finding was that the application of {PSS/PAH} films as thin as 12 nm can drastically improve the sensor performance over uncoated sensors based on calcium alginate microspheres. The model is proposed as an important tool for a priori design of these complex sensor structures. PMID- 16219458 TI - Transposable elements, gene creation and genome rearrangement in flowering plants. AB - Plant genome structure is largely derived from the differing specificities, abundances and activities of transposable elements. Recent studies indicate that both the amplification and the removal of transposons are rapid processes in plants, accounting for the general lack of intergenic homology between species that last shared a common ancestor more than 10 million years ago. Two newly discovered transposon varieties, Helitrons and Pack-MULEs, acquire and fuse fragments of plant genes, creating the raw material for the evolution of new genes and new genetic functions. Many of these recently assembled, chimeric gene candidates are expressed, suggesting that some might escape epigenetic silencing and mutational decay, but a proven case of gene creation by any transposable element activity in plants remains to be demonstrated. PMID- 16219459 TI - Expression of JC virus regulatory proteins in human cancer: potential mechanisms for tumourigenesis. AB - JC virus (JCV) is a human polyomavirus that is the etiologic agent of the fatal demyelinating disease of the central nervous system known as progressive multifocal leukoencephalopathy (PML). JCV is also linked to some tumours of the brain and other organs as evidenced by the presence of JCV DNA sequences and the expression of viral proteins in clinical samples. Since JCV is highly oncogenic in experimental animals and transforms cells in culture, it is possible that JCV contributes to the malignant phenotype of human tumours with which it is associated. JCV encodes three non-capsid regulatory proteins: large T-antigen, small t-antigen and agnoprotein that interact with a number of cellular target proteins and interfere with certain normal cellular functions. In this review, we discuss how JCV proteins deregulate signalling pathways especially ones pertaining to transcriptional regulation and cell cycle control. These effects may be involved in the progression of JCV-associated tumours and may represent potential therapeutic targets. PMID- 16219460 TI - Determining the modern carbon content of biobased products using radiocarbon analysis. AB - In support of the USDA Federal Biobased Products Preferred Procurement Program (FB4P), Iowa State University is coordinating testing to determine the "biobased content" of manufactured products. These tests are part of a process to "designate" items that qualify for preferential procurement status with federal agencies. Biobased content determinations are being performed using three radiocarbon dating procedures specified in ASTM D 6866-05. Test results obtained thus far indicate that the AMS and benzene synthesis methods provide comparable results. Data from the CO(2) cocktail method did not agree as well with the data from the other two methods, but were still in reasonably good agreement with those data. Radiocarbon analysis is shown to be a reliable and valuable tool for verifying the biobased content of a wide variety of biobased products. Based on inter- and intra-laboratory comparisons, a reasonable uncertainty to associate with the analyses would be +/-3% (absolute) for the AMS and benzene synthesis methods. Because of limited data availability, additional work is needed to establish the uncertainty of the CO(2) cocktail method for analyzing biobased products. PMID- 16219461 TI - The flocculation performance of Tamarindus mucilage in relation to removal of vat and direct dyes. AB - A food grade natural mucilage, extracted from the seeds of Tamarindus indica pods, is used as a flocculant for removal of solubilised vat (golden yellow) and direct dye (direct fast scarlet) in aqueous solutions. The maximum removal obtained was 60% for golden yellow after 2 h and was 25% for direct fast scarlet after 1 h. The optimum mucilage dose was 10 mg/l and 15 mg/l for golden yellow and direct fast scarlet, respectively. The pH values also seem to affect the percent removal of both the dyes significantly. In case of vat dye, the pH value of the test samples affected the percent removal significantly. The change was highly significant between neutral and alkaline pH. In case of direct dye, there was no significant change in percent removal at pH 7 and pH 4 whereas a significant change in percent removal was observed between pH 7 and pH 9.2. The plausible mucilage-dye interaction and flocculation mechanism has been discussed. This new flocculant works better in the case of vat dye removal compared with the direct dye. PMID- 16219462 TI - Mercury(II) removal from aqueous solutions by nonviable Bacillus sp. from a tropical estuary. AB - Use of microorganisms for removing mercury is an effective technology for the treatment of industrial wastewaters and can become an effective tool for the remediation of man-impacted coastal ecosystems with this metal. Nonviable biomass of an estuarine Bacillus sp. was employed for adsorbing Hg(II) ions from aqueous solutions at six different concentrations. It was observed that 0.2 g dry weight of nonviable biomass was found to remove from 0.023 mg (at 0.25 mg L(-1) of Hg(II)) to 0.681 mg (at 10.0 mg L(-1) of Hg(II)). Most of the mercury adsorption occurred during the first 20 min. It was found that changes in pH have a significant effect on the metal adsorption capacity of the bacteria, with the optimal pH value between 4.5 and 6.0 at 25 degrees C when solutions with 1.0, 5.0 and 10.0 mg L(-1) of Hg(II) were used. PMID- 16219463 TI - Estrogen receptor beta selective ligands: discovery and SAR of novel heterocyclic ligands. AB - A series of ligands with varying heterocyclic cores and substituents that display a range of selectivity's (up to >100x) for ER-beta over ER-alpha are reported. PMID- 16219464 TI - Metalloform-selective inhibition: synthesis and structure-activity analysis of Mn(II)-form-selective inhibitors of Escherichia coli methionine aminopeptidase. AB - Methionine aminopeptidase (MetAP) is a promising target for development of novel antibacterial, antifungal and anticancer agents. However, its physiologically relevant metal ion remains to be defined, and its inhibitors need to inhibit the in vivo metalloform. Based on the Mn(II)-form-selective inhibitors discovered by high throughput screening as leads, a series of analogs of 5-phenylfuran-2 carboxylic acid was prepared and subsequently evaluated on Co(II)-, Mn(II)-, Ni(II)-, and Fe(II)-forms of Escherichia coli MetAP, in order to define the structural elements responsible for their inhibitory potency and metalloform selectivity. Various substitutions on the phenyl ring changed their potency on the Mn(II)-form but not their metalloform selectivity. We conclude that the preferential coordination of the carboxyl group to Mn(II) ions is the major determinant for their superb selectivity toward the Mn(II)-form. Changing the carboxylate to hydroxamate alters its ability to bind and discriminate different metal ions, and the hydroxamate derivative becomes non-selective among the metalloforms tested. PMID- 16219465 TI - Synthesis and SAR of highly potent dual 5-HT1A and 5-HT1B antagonists as potential antidepressant drugs. AB - Novel 5-HT(1) autoreceptor ligands based on the N-4-aryl-piperazinyl-N'-ethyl 5,6,7,8-tetrahydropyrido[4', 3':4,5]thieno[2,3-d]pyrimidin-4(3H)-one core are described. Aiming at antidepressants with a novel mode of action our objective was to identify potent antagonists showing balanced affinities and high selectivity for the 5-HT(1A) and 5-HT(1B) receptors. Strategies for the development of dual 5-HT(1A) and 5-HT(1B) antagonists based on 1 and 2 as leads and the corresponding results are discussed. Isoquinoline analogue 33 displayed high affinity and an antagonistic mode of action for the 5-HT(1A) and the 5 HT(1B) receptors and was characterized further with respect to selectivity, electrically stimulated [(3)H]5-HT release and in vivo efficacy. PMID- 16219466 TI - Structure activity studies of the serine-AIB dipeptide domain in 2,3 dihydroisothiazole based growth hormone secretagogues. AB - A series of growth hormone secretagogues (GHSs) based on 2,3-dihydroisothiazole has been synthesized in the search for a potential treatment of growth hormone deficiency or frailty in the elderly. This paper describes the evaluation of the SAR of the benzyl-D-Ser-aminoisobutyric acid dipeptide fragment. Introduction of substituents in the peptide backbone and in the phenyl ring has been investigated, as well as replacements for the benzyl group and for the AIB residue. A number of modifications resulted in enhanced potency over the parent benzyl-D-Ser-AIB derivative. PMID- 16219468 TI - Enaminones 9. Further studies on the anticonvulsant activity and potential type IV phosphodiesterase inhibitory activity of substituted vinylic benzamides. AB - Structure-activity relationship studies were employed to synthesize a series of 3 and 3,4-substituted benzamides from 3-amino-2-cyclohexenones. An improved method for the synthesis of benzamides from 3-amino-2-cyclohexenones is presented which provided significantly higher yields (71-79%) for the reported compounds. NMR and X-ray structural analyses were undertaken to note the possible intra- and intermolecular interactions of the synthesized analogs. Molecular modeling studies were used to determine the minimized configuration and were compared to their X-ray structures for correlation. These new entities were evaluated as potential anticonvulsants and type IV phosphodiesterase inhibitors (PDE4). PMID- 16219467 TI - Synthesis, radiosynthesis and in vivo preliminary evaluation of [11C]LBT-999, a selective radioligand for the visualisation of the dopamine transporter with PET. AB - LBT-999 (8-((E)-4-fluoro-but-2-enyl)-3beta-p-tolyl-8-aza-bicyclo[3.2.1]octane 2beta-carboxylic acid methyl ester), a cocaine derivative belonging to a new generation of highly selective dopamine transporter (DAT) ligands, and its corresponding carboxylic acid derivative, the latter used as precursor for labelling both with tritium and the positron-emitter carbon-11 (half-life: 20.38 min), were synthesized from (R)-cocaine. [(3)H]LBT-999 (>99% radiochemically pure, specific radioactivity of 3.1 TBq/mmol) was prepared from [(3)H]methyl iodide, allowing its in vitro pharmacological evaluation (K(D): 9 nM for DAT and IC(50) > 1000 nM for SERT and NET). Routine production batches of 4.5-9.0 GBq of iv injectable solutions of [(11)C]LBT-999 (with specific radioactivities ranging from 30 to 45 GBq/mumol) were prepared in 25-30 min (HPLC purification and formulation included) using the efficient methylation reagent [(11)C]methyl triflate. The preliminary in vivo pharmacological evaluation of [(11)C]LBT-999, using both biodistributions in rats and brain imaging in monkeys with positron emission tomography (PET), clearly illustrates that this ligand is an excellent candidate for quantification with PET of DAT in humans. PMID- 16219469 TI - The influence of the halide ions on the photochemical reaction cycle of pharaonis halorhodopsin. AB - Sodium salt of chloride, bromide and iodide were used to elucidate the effect of the size of the anion on the binding to pharaonis halorhodopsin and its transport during the photocycle of this retinal protein. Spectroscopic titration revealed an apparent strong binding constant of 2 mM for chloride, 0.23 mM for bromide and 5 mM for iodide. In the case of iodide a second, week binding constant of about 10 M could be estimated. This second binding constant was similar to that observed earlier for nitrate. By changing the halide ions, only the transitions in the second half of the photocycle were affected, which contained intermediates N, O, and HR'. The O to HR' transition becomes faster with increasing ion volume, meaning that the ion uptake is accelerated. This effect shows a direct correlation with the ion radius. With increasing ion concentration the N-O-HR' equilibrium changed in such a way that the accumulated O tended to decrease. This tendency was overruled in iodide, by the appearance of the second binding constant. The increasing iodide concentration, up to 100mM decreases the accumulation of the intermediate O, due to kinetic reasons, but at higher ion concentration the amount of O increases, although its decay becomes faster. This effect correlates with the appearance of the second iodide bound to the protein. PMID- 16219470 TI - Effects of solar radiation on collagen and chitosan films. AB - Photo-aging and photo-degradation are the deleterious effect of chronic exposure to sun light of many materials made of natural polymers. The resistance of the products on the action of solar radiation is very important for material scientists. The effect of solar radiation on two natural polymers: collagen and chitosan as well as collagen/chitosan blends in the form of thin films has been studied by UV-Vis and FTIR spectroscopy. It was found that UV-Vis spectra, which characterise collagen and collagen/chitosan films, were significantly altered by solar radiation. FTIR spectra of collagen and collagen/chitosan films showed that after solar irradiation the positions of amide A and amide I bands were shifted to lower wavenumbers. There was not any significant alteration of chitosan UV-Vis and FTIR spectra after solar radiation. In the condition of the experiment chitosan films were resistant to the action of solar radiation. The effect of solar UV radiation in comparison to artificial UV radiation has been discussed. PMID- 16219471 TI - Self-assembly of meso-tetrakis(4-N-ethylpyridiniurmyl) porphyrin and tetracarboxyl-phenyl calix[4]arene. AB - Novel dimeric capsules are generated from the noncovalent assembly of 5, 10, 15, 20-tetrakis(4-N-ethylpyridiniurmyl)prophyrin (TEPyP) and tetracarboxyl-phenyl calix[4]arene. The self-assembly system was investigated based on UV-Vis absorption and fluorescent spectra. The factors affecting the interaction process including pH and concentration were examined in detail. The association constants between TEPyP and calix[4]arene were determined by the nonlinear least squares fit. The results showed that the basic medium is favourable to the interaction and electrostatic interaction was determinate in the processes of self-assembly process. The related mechanism was discussed. PMID- 16219472 TI - Expression of icb-1 gene is interferon-gamma inducible in breast and ovarian cancer cell lines and affects the IFN gamma-response of SK-OV-3 ovarian cancer cells. AB - Icb-1 (C1orf38) is a human gene initially described to be involved in in vitro differentiation processes of endometrial adenocarcinoma and leukemia cells. In this study, we examined the effect of interferon-gamma on icb-1alpha and beta mRNA levels in human cell lines derived from breast cancer and gynecological malignancies. Furthermore, we intended to approach icb-1 gene function by means of RNA interference (RNAi) to analyze the effect of an icb-1 knockdown on human cancer cells in vitro. Three breast cancer cell lines (MCF-7, SK-BR-3, MDA-MB 231), three ovarian cancer cell lines (SK-OV-3, OVCAR-3 and BG-1) and the endometrial adenocarcinoma cell line HEC-1-A were treated with interferon gamma and the transcript levels of icb-1 isoforms alpha and beta were assessed by means of semiquantitative real-time RT-PCR. Our data demonstrates an interferon-gamma triggered upregulation of icb-1alpha mRNA levels in all breast cancer cell lines and an increase of icb-1beta mRNA in MDA-MB-231 cells. The strongest (about 10 fold) increase of icb-1alpha and beta mRNA after treatment with interferon-gamma was observed in ovarian cancer cell line SK-OV-3. Additionally, our data demonstrates the success of a siRNA-mediated knockdown of icb-1alpha and beta mRNA levels, which resulted in a significant increase of the antiproliferative interferon-gamma effect on SK-OV-3 cells. In conclusion, we report identification of the novel interferon-gamma inducible gene icb-1 which is able to affect the response of ovarian cancer cells to this cytokine. PMID- 16219473 TI - Mechanisms of T cell motility and arrest: deciphering the relationship between intra- and extracellular determinants. AB - T lymphocytes are capable of rapid motility in vitro and in vivo. Upon antigen recognition, they may stop crawling and form a stable cell-cell contact called the 'immunological synapse' (IS). However, it is becoming clear that this outcome may not occur with the reliability that was once presumed. T cells, particularly naive cells, are apparently triggered partly 'on the fly' during short contacts with peptide-MHC (pMHC) bearing antigen-presenting cells (APCs) and are also influenced in both activity and synapse duration by a multitude of external cues. Underlying the emerging issues is a paucity of data concerning the cell biology of T lymphocytes. Here, we review the molecular mechanisms of crawling and adhesion versus the various potential modes of 'stopping' in T lymphocytes. Both motility and arrest involve similar processes: adhesion, actin elongation and internal tension control, but with different coordination. We will attempt to integrate this with the known and potential external cues that signal for T cell motility versus stopping to form a synapse in vivo. Finally, we discuss how this interplay may give rise to unexpectedly complex motile and morphological behavior. PMID- 16219474 TI - Activation of the p38MAPK cascade is associated with upregulation of TNF alpha receptors in the spinal motor neurons of mouse models of familial ALS. AB - Phosphorylated p38 mitogen-activated protein kinase (p38MAPK), but not activated c-jun-N-terminal kinase (JNK), increases in the motor neurons of transgenic mice overexpressing ALS-linked SOD1 mutants at different stages of the disease. This effect is associated with a selective increase of phosphorylated MKK3-6, MKK4 and ASK1 and a concomitant upregulation of the TNFalpha receptors (TNFR1 and TNFR2), but not IL1beta and Fas receptors. Activation of both p38 MAPK and JNK occurs in the activated microglial cells of SOD1 mutant mice at the advanced stage of the disease; however, this effect is not accompanied by the concomitant activation of the upstream kinases ASK1 and MKK3,4,6, while both the TNFRs are overexpressed in these cells. No changes of the upstream p38MAPK cascade kinases or TNFRs occur in reactive astrocytes. These findings highlight the activation of a selective intracellular signaling pathway in the motor neurons of SOD1 mutant mice, which is likely implicated in their death. PMID- 16219475 TI - Influence of high-M alginate on the growth and survival of Atlantic cod (Gadus morhua L.) and spotted wolffish (Anarhichas minor Olafsen) fry. AB - Atlantic cod and spotted wolffish fry were fed high-M alginate containing feed for 59 and 55 days, respectively. During this period the fry showed a higher specific growth rate compared to controls. Uptake and distribution of alginate was studied by inclusion of the (125)I-labelled molecule in the feed. The stomach and intestine contained the highest amount while the kidney, liver and spleen contained some, indicating that the alginate was taken up by the gut and transported to internal organs. Cod fry fed 0.06% and 0.1% high-M alginate showed a death rate of 51.4% and 53.3%, respectively. The lowest mortality, 48.1%, was found in fry fed 0.01% high-M alginate. Controls showed a mortality rate of 49.0%. Differences were, however, not statistically significant. Challenge of the immunostimulated fry (fed 0.02% and 0.06% alginate for 62 days) with atypical Aeromonas salmonicida bacteria resulted in accumulated mortalities of 56% and 49%, respectively, 47 days after infection. The group that received 0.06% alginate for a shorter period (47 days) and then control feed until challenged, and the group that received alginate by bath reached a cumulative mortality of 59% and 60%, respectively. Lowest mortality (44%) was seen in the control group. Numerous microabscesses were found in both immunostimulated and control fish in secondary lamellae of the gills, haematopoietic tissues of the kidneys, the submucosa and mucosa of the intestine, the spleen, the liver and the myocardium of the heart. PMID- 16219476 TI - Trichlorfon, an organophosphorus insecticide, depresses the immune responses and resistance to Lactococcus garvieae of the giant freshwater prawn Macrobrachium rosenbergii. AB - The total haemocyte count (THC), phenoloxidase activity, respiratory bursts (release of superoxide anions), superoxide dismutase (SOD) activity, and phagocytic activity and clearance efficiency to the pathogen Lactococcus garvieae were measured when freshwater giant prawns Macrobrachium rosenbergii (18.2+/-2.1 g) were individually reared in water containing concentrations of trichlorfon of 0, 0.2, or 0.4 mg L(-1) for a 144-h period. No significant differences in the THC were observed among prawns at the beginning and following 144 h of exposure to 0, 0.2, and 0.4 mg L(-1) trichlorfon. However, phenoloxidase activity significantly decreased when the prawns were exposed to trichlorfon at 0.2 and 0.4 mg L(-1), and no significant differences were observed between the two concentrations at any sampling time. The total production of superoxide anion by M. rosenbergii significantly increased with exposure to 0.2 and 0.4 mg L(-1) trichlorfon, and no significant differences were observed between the two concentrations during the 144-h exposure period. However, M. rosenbergii exposed to 0.2 and 0.4 mg L(-1) trichlorfon showed decreased SOD activity from 48 to 144 h. Phagocytic activity and clearance efficiency to L. garvieae significantly decreased when prawns were exposed to 0.2 and 0.4 mg L(-1) trichlorfon for 48 h. In another experiment, prawns were challenged with 5 x 10(5) colony-forming units (cfu)prawn(-1) of L. garvieae, then reared in water containing different concentrations of trichlorfon. The onset of mortality was earlier in prawns exposed to trichlorfon compared to those exposed to the zero control. The cumulative mortality of prawns directly increased with ambient trichlorfon concentrations in the range of 0-0.4 mg L(-1) after 168 h. It is concluded that exposure of M. rosenbergii to trichlorfon at 0.2 mg L(-1) or more caused cytotoxicity resulting in depression of the immune response, and increased its susceptibility to L. garvieae infection. PMID- 16219477 TI - Ballistic muscle mechanisms determined using an EMG-driven model. AB - The purpose of this study was to determine the mechanisms responsible for improving ballistic elbow extension. In doing so, an electromyography (EMG) driven model was developed to predict the actual triceps torque so that the model parameters between subjects could be compared. Thirty-two subjects performed maximum isometric trials at 60 degrees , 90 degrees and 120 degrees of elbow extension to determine torque-angle relations. Dynamic elbow extension trials were then performed against relative loads of 0%, 20%, 40%, 60% and 80% and absolute loads of 1.1 and 2.2 kg. These trials were used to determine the torque angular velocity relation for each subject. The model predicted the triceps torque during the unloaded, 1.1 and 2.2 kg trials with an average r = 0.964 and an average root mean square error of 4.34 Nm. As a result of the good predictions, a forward dynamics approach was used to substitute different neuro muscular mechanisms of a poor performance with those from an individual that displayed a superior performance. Performance was shown to improve when these modifications were made. Therefore, the EMG-driven model was capable of modeling the actual muscle torque which allowed for the identification of areas of weakness of a poor performance. A prescription for improvement was identified, albeit artificially, on an individual basis. The next stage is to determine which specific interventions can accomplish those theoretically proposed. PMID- 16219478 TI - [Radiation therapy of benign diseases. What's new eight years after?]. AB - The authors present an update version of the indications for radiotherapy in the management of benign diseases. This is based on available randomized trials and recent international meetings. Validated indications remain the prevention of resected heterotopic bone ossifications, keloids scars and pterygium and also treatment of arteriovenous malformations; the place of radiotherapy for malignant exophtalmia is more and more restricted. Randomized trials have demonstrated the efficacy of endobrachytherapy in the prevention of restenosis after angioplasty but the use of embedded stent has replaced this indication. Macular degeneration is no more an indication of radiotherapy. Quality requirements for radiotherapy are identical for benign or malignant indications. PMID- 16219479 TI - [Recent progress in treatment of malignant pleural mesothelioma]. AB - Incidence of malignant pleural mesothelioma will rise until 2030-2040 because the elapsed time between exposure and diagnostic is up to several decades. Prognosis remains very poor with median survival less than one year and five-year survival not exceeding 5%. As compared to 1999, standart treatment adds chemotherapy with cisplatin and pemetrexed to local radiotherapy for prevention of local seeding after invasive diagnostic procedures. Despite various growth factors and their receptors are involved in malignant mesothelioma, first clinical trials of targeted therapies reported poor results. Multimodality therapy with extrapleural pneumonectomy and radiation therapy (+/-chemotherapy) can be of benefit in subgroups of patients but it cannot be recommended in a routine approach. As compared to bronchial carcinoma, inclusion of patients in clinical trials (using intensity-modulated radiation therapy) is the only way to somewhat improve results. PMID- 16219480 TI - [Tolerance and efficacy of conformal radiotherapy for hepatocellular carcinoma in cirrhotic patients. Results of the French RTF1 phase II trial]. AB - PURPOSE: While some patients presenting with hepatocellular carcinoma (HCC) benefit from curative therapies (transplantation, surgery, percutaneous ablation), others are only candidates for palliative options such as chemoembolization or symptomatic care. Although conventional external-beam radiotherapy of the liver is regarded as little efficient and potentially toxic in cirrhotic patients, 3-dimensional conformal radiotherapy (CRT), by decreasing the amount of normal liver included in the radiation portal, allows dose escalation to occur without increasing the risk of radiation-induced hepatitis. This trial was designed to assess the efficacy and tolerance of CRT for small size HCC in cirrhotic patients. PATIENTS AND METHODS: Prospective phase II trial including stage A/B cirrhotic patients with small-size HCC not suitable for curative treatments; CRT consisted in a standard fractionation radiation, with a total dose of 66 Gy. RESULTS: Twenty-seven patients were included, 15 of whom had previously been treated for HCC; mean age was 68. Among the 23 assessable patients, 18 (78%) presented with complete response, 3 (13%) with partial response, and 2 with no response. Acute complications occurred in 24 patients, and were mainly acceptable (grade 1/2: 22 patients, grade 3/4: 11 patients, 4 (15%) of whom had clinical and/or hematological toxicities). Only 2 (9%) grade 3/4 clinical and/or hematological late toxicities are reported. CONCLUSION: CRT is a non-invasive curative technique highly suitable for small-size HCC in cirrhotic patients; further investigations are needed to compare it to the other available treatments, and to integrate it into the curative therapeutic algorithm of HCC. PMID- 16219481 TI - [Economic evaluation of radiotherapy: methods and results]. AB - The cost of radiotherapy, which concerns 150,000 new patients yearly in France has been estimated 1700 euros per patients in this country, 50% lower than the European average. Economic evaluations of radiotherapy have related the cost of either curative or palliative treatment to the medical outcome. For lung, rectal, breast cancer and bone metastases of prostate cancer, radiation therapy prolongs life by an estimated 6 to 10 months. The cost effectiveness ranges from 5,000 to 25,000 euros per life year gained. PMID- 16219482 TI - Fatal traumatic asphyxia in a middle-aged man in association with entrapment associated hypoxyphilia. AB - A case is reported of a 36-year-old man who died following occlusive entrapment within a device for the purpose of hypoxyphilic gratification. The device was constructed in his own home using instructions found on his home computer down loaded from the Internet. The device comprised a tough plastic cocoon large enough to accommodate an adult human and incorporating a system of plastic piping connected to a household vacuum cleaner for the evacuation of air within the cocoon. The mechanism of death was thought to be traumatic asphyxia after examination of the deceased and re-construction of the apparatus with the body in situ. The implications of this unusual device for the purposes of hypoxyphilic gratification by occlusive entrapment and its inherent dangers are highlighted. PMID- 16219483 TI - Melatonin. AB - Melatonin, originally discovered as a hormone of the pineal gland, is produced by bacteria, protozoa, plants, fungi, invertebrates, and various extrapineal sites of vertebrates, including gut, skin, Harderian gland, and leukocytes. Biosynthetic pathways seem to be identical. Actions are pleiotropic, mediated by membrane and nuclear receptors, other binding sites or chemical interactions. Melatonin regulates the sleep/wake cycle, other circadian and seasonal rhythms, and acts as an immunostimulator and cytoprotective agent. Circulating melatonin is mostly 6-hydroxylated by hepatic P450 monooxygenases and excreted as 6 sulfatoxymelatonin. Pyrrole-ring cleavage is of higher importance in other tissues, especially the brain. The product, N1-acetyl-N2-formyl-5 methoxykynuramine, is formed by enzymatic, pseudoenzymatic, photocatalytic, and numerous free-radical reactions. Additional metabolites result from hydroxylation and nitrosation. The secondary metabolite, N1-acetyl-5-methoxykynuramine, supports mitochondrial function and downregulates cyclooxygenase 2. Antioxidative protection, safeguarding of mitochondrial electron flux, and in particular, neuroprotection, have been demonstrated in many experimental systems. Findings are encouraging to use melatonin as a sleep promoter and in preventing progression of neurodegenerative diseases. PMID- 16219484 TI - Genotoxic risk and oxidative DNA damage in HepG2 cells exposed to perfluorooctanoic acid. AB - Perfluorooctanoic acid (C8HF15O2, PFOA) is widely used in various industrial fields for decades and it is environmentally bioaccumulative. PFOA is known as a potent hepatocarcinogen in rodents. But it is not yet clear whether it is also carcinogenic in humans, and the genotoxic effects of PFOA on human cells have not yet been examined. In this study, the genotoxic potential of PFOA was investigated in human hepatoma HepG2 cells in culture using single cell gel electrophoresis (SCGE) assay and micronucleus (MN) assay. In order to clarify the underlying mechanism(s) we measured the intracellular generation of reactive oxygen species (ROS) using dichlorofluorescein diacetate as a fluorochrome. The level of oxidative DNA damage was evaluated by immunocytochemical analysis of 8 hydroxydeoxyguanosine (8-OHdG) in PFOA-treated HepG2 cells. PFOA at 50-400 microM caused DNA strand breaks and at 100-400 microM MN in HepG2 cells both in a dose dependent manner. Significantly increased levels of ROS and 8-OHdG were observed in these cells. We conclude that PFOA exerts genotoxic effects on HepG2 cells, probably through oxidative DNA damage induced by intracellular ROS. PMID- 16219485 TI - Methodology to ease the construction of a terminology of problems. AB - INTRODUCTION: Problem lists summarize an aspect of the patient's medical history and provide an important way to implement entry points for clinical pathways and guideline-oriented care. However, in order to automate processes based on problem lists, the use of controlled vocabularies is required. We developed a methodology to extract a collection of standardized problem-related terms from medical documents entered in free text by physicians. METHODS: We extracted a corpus of sentences describing problems from a randomized selection of admission notes collected at the University Hospitals of Geneva. Theses sentences underwent manual and automatic normalization processes, and a statistical clustering, in order to build a set of terms. RESULTS: We obtained 17,805 sentences from 5000 admission notes. We refined them into 1546 terms, 88.6% of which could be related to a relevant problem statement. DISCUSSION: A clinically relevant problems terminology was derived from clinical admission notes in free-text using a few methodical steps with a reasonable investment of human resources. Such an approach will ease the development and the use of problem lists better suited to user needs. PMID- 16219486 TI - Histological study on comparison between NASH and ALD. AB - We examined the histological findings of non-alcoholic steatohepatitis (NASH) and ALD to study resemblance and difference of these two diseases. At a glance of H&E staining, they looked same, however, careful examination on silver impregnation histology, the fibrosis of NASH showed lattice fibrosis on the other hand ALD showed solid fibrosis. The appearance of fibrosis was different due to qualitative difference. More examination on the change of hepatocytes, there were some difference in incidence of the findings such as megamitochondria, bile stasis, hemosiderin deposition, vacuolic nuclei, and lipogranuroma. So we are able to differentiate NASH and ALD histologically. PMID- 16219487 TI - P300 after head injury: pseudodelay caused by reduced P3A amplitude. AB - OBJECTIVE: We compared conventional P300 analysis with source analysis in normal subjects and head-injury patients. Based on earlier findings of improved P300 component identification and reduced P3B latency variability with source analysis in normal subjects, our aim was to investigate whether source analysis could improve the distinction between these groups. METHODS: In total, 21 healthy control subjects and 21 patients with mild to moderate head injury were included in this study. A standard auditory 2-tone oddball paradigm was used. Latencies and amplitudes obtained with conventional P300 analysis were compared with source analysis results. RESULTS: With conventional analysis, head-injury patients had delayed P300 latencies and reduced P300 amplitudes in comparison to controls, while source analysis showed no latency differences for both P3A and P3B components. Instead, source analysis indicated absence of P3A components in 43% of patients. CONCLUSIONS: The P300 delay in head-injury patients, observed with conventional analysis, is a pseudodelay caused by decreased P3A amplitudes. Consequently, the unaffected P3B component with its later latency determines conventional P300 latency in these patients. SIGNIFICANCE: Conventional P300 latency cannot be used to conclude that there was delayed early stimulus processing in head-injury patients. PMID- 16219488 TI - Comments on selected fundamental aspects of microarray analysis. AB - Microarrays are becoming a ubiquitous tool of research in life sciences. However, the working principles of microarray-based methodologies are often misunderstood or apparently ignored by the researchers who actually perform and interpret experiments. This in turn seems to lead to a common over-expectation regarding the explanatory and/or knowledge-generating power of microarray analyses. In this note we intend to explain basic principles of five (5) major groups of analytical techniques used in studies of microarray data and their interpretation: the principal component analysis (PCA), the independent component analysis (ICA), the t-test, the analysis of variance (ANOVA), and self organizing maps (SOM). We discuss answers to selected practical questions related to the analysis of microarray data. We also take a closer look at the experimental setup and the rules, which have to be observed in order to exploit microarrays efficiently. Finally, we discuss in detail the scope and limitations of microarray-based methods. We emphasize the fact that no amount of statistical analysis can compensate for (or replace) a well thought through experimental setup. We conclude that microarrays are indeed useful tools in life sciences but by no means should they be expected to generate complete answers to complex biological questions. We argue that even well posed questions, formulated within a microarray-specific terminology, cannot be completely answered with the use of microarray analyses alone. PMID- 16219489 TI - A family of metrics for biopolymers based on counting independent sets. AB - We introduce a new family of metrics for graphs of fixed size, based on counting independent sets. Our definition is simpler and easier to calculate than the edge ideal metric family defined by Llabres and Rossello without loosing any of its abstract properties. We contrast them on some examples with graphs that represent protein secondary and three-dimensional (3D) structures. We conclude that although the edge ideal metrics are faster to calculate on some sparse graphs, in general, the independent set metrics are more tractable. PMID- 16219490 TI - Transfection of dendritic cells with in vitro-transcribed CMV RNA induces polyclonal CD8+- and CD4+-mediated CMV-specific T cell responses. AB - Transfection of dendritic cells (DCs) with RNA was shown to be effective in the generation of antigen-specific T cells, probably due to the induction of a polyclonal T cell response directed against multiple antigens. To verify this assumption we used DCs, generated from cytomegalovirus (CMV)-negative or positive donors, that were electroporated with in vitro-transcribed RNA (in vitro transcript, IVT) coding for the CMV pp65 antigen. We found that transfection of DCs with pp65 IVT induces an expansion of polyclonal CD8(+) T lymphocytes that recognize peptide antigens presented on different HLA molecules. These T lymphocytes are able to lyse DCs pulsed with pp65-derived peptides or transfected with the cognate IVT. Furthermore, this approach allowed the identification of immunodominant epitopes presented upon IVT transfection. Interestingly, transfection of DCs with pp65 IVT resulted in the induction of CD4(+)-specific T cells. Cotransfection of DCs with IVTs coding for the CMV antigens pp65 and IE1 elicited polyclonal T lymphocytes specific for peptides derived from both antigens. More importantly, cytotoxic T cells could be generated in two of three CMV-negative donors. Finally, functional CMV-specific autologous cytotoxic T lymphocytes were successfully generated from immunosuppressed patients after allogeneic hematopoietic stem cell transplantation. PMID- 16219491 TI - Bone marrow transplantation combined with gene therapy to induce antigen-specific tolerance and ameliorate EAE. AB - Hematopoietic stem cell (HSC) transplantation is a potential therapy that can offer multiple sclerosis patients a radical, potentially curative treatment. Using experimental autoimmune encephalomyelitis (EAE) as a model, we previously reported that retrovirally transduced B cells expressing myelin basic protein (MBP), MBP Ac1-11, or myelin oligodendrocyte glycoprotein p35-55 induced tolerance and reduced symptoms. Here, we extend our tolerance approach using bone marrow (BM) cells expressing full-length phospholipid protein (PLP) in a model for relapsing, remitting EAE. Using GFP expression as a marker, we found that up to 50% of cells were positive for transgene expression in peripheral blood after 900 rad irradiation and transduced BM transplantation, and expression was stable in hematopoietic lineages for over 10 weeks. Upon challenge, T cell proliferation in response to PLP p139-151 was reduced and EAE was completely abolished in a pretreatment protocol. In addition, protection from EAE could be achieved with PLP-transduced BM cells given on day 12 after immunization, a potential therapeutic protocol. Finally, the protective effect of PLP-expressing BM could also be observed using a nonmyeloablative protocol, albeit with lower efficacy. Our results suggest that HSC may be useful to achieve long-lasting tolerance to protect mice from EAE and possibly to promote CNS repair in ongoing EAE. PMID- 16219492 TI - Biology of AAV serotype vectors in liver-directed gene transfer to nonhuman primates. AB - Vectors based on adeno-associated viruses (AAVs) show promise for the treatment of genetic diseases. This study evaluates the biology of AAV-mediated gene transfer to liver in nonhuman primates (NHPs) using vectors based on AAV serotypes 2, 7, and 8. Transgenes encoding self-proteins were selected to minimize the confounding development of transgene-specific immune responses. These included the beta subunit of choriogonadotropic hormone (bCG) and erythropoietin (Epo), both derived from cDNAs from rhesus macaques. Experiments were performed with bCG in rhesus macaques and Epo in cynomolgus macaques. We demonstrated the previously untested hypothesis that preexisting immunity to a natural infection does substantially diminish the efficacy of gene transfer with a vector derived from an endogenous virus. Route of vector administration clearly has an impact on the development of immune responses to self-antigens. In general, efficiency of gene transfer to liver with AAV7 and 8 vectors was higher than what was achieved with AAV2, although a variety of host factors may influence this important parameter, such as preexisting immunity, gender, and transgene immunity. PMID- 16219494 TI - Quantification of NC100668, a new tracer for imaging of venous thromboembolism, in human plasma using reversed-phase liquid chromatography coupled with electrospray ionization ion-trap mass spectrometry. AB - NC100668 is being developed as a new tracer for radiopharmaceutical imaging of venous thromboembolism. NC100668 consists of a targeting peptide of 13 amino acids with a (99m)Tc-binding chelator linked to the C-terminal amino acid. The present report describes a method for quantification of NC100668 in human citrated plasma. The method is based on solid-phase extraction followed by reversed-phase liquid chromatography using a gradient of water and acetonitrile with 0.1% formic acid. The chromatographic system was coupled on-line with an electrospray mass spectrometer. The analyses were performed by selective ion monitoring of the [M+2H]2+ and the [M+3H]3+ ions of NC100668 and an internal standard which was identical to NC100668 except for not being iodinated in the tyrosine residue. The limit of quantification of the method was 2ng NC100668/ml plasma. The calibration curve ranged from 2 to 250 ng NC100668/ml plasma and was fitted to a linear equation with a weighing factor of 1/y2 and found to be highly reproducible. The total precision of the method, expressed as the relative standard error of the mean, was 23.2, 8.8 and 14.7% for the low, medium and high control samples, respectively. The accuracy of the method was 108.5, 100.0 and 105.0% for the low, medium and high control samples, respectively. NC100668 was stable in human plasma during at least three freeze/thaw cycles, during 30 h on dry ice and up to 3 months when stored in a -20 degrees C freezer. PMID- 16219493 TI - Tissue harmonic imaging for standard left ventricular measurements: fundamentally flawed? AB - Tissue harmonic imaging (THI) is a B mode imaging technique that improves echocardiographic image quality by reducing superficial artefact. The modality increases image signal-to-noise ratio at the expense of reduced axial resolution. While the qualitative improvements of harmonic echocardiographic imaging are widely accepted, the degree to which this is translated into improved quantitative measurements and whether THI-derived measurements result in systematic bias continue to be areas of uncertainty. This review examines differences between THI and fundamental imaging-derived measurements from a theoretical, tissue phantom and clinical perspective. PMID- 16219495 TI - Kinetic characterization of Synechocystis sp. PCC6803 1-deoxy-D-xylulose 5 phosphate reductoisomerase mutants. AB - The methylerythritol phosphate pathway to isoprenoids has been firmly established as an alternate to the mevalonate pathway in many bacteria, plants, algae, and the malaria parasite Plasmodium falciparum. The second enzyme in this pathway, deoxy-D-xylulose 5-phosphate reductoisomerase (DXR; E.C. 1.1.1.267), has been the focus of many investigations since it was found to be the target of the antibacterial and antimalarial compound, fosmidomycin. Several x-ray crystal structures of the Escherichia coli and Zymomonas mobilis DXR enzymes have provided important structural information about the residues potentially involved in substrate binding and catalysis. Site-directed mutagenesis studies can be used to complement the structural studies, providing kinetic data for specific changes of active site residues. Active site mutants were prepared of the recombinant Synechocystis sp. PCC6803 DXR, targeting residues D152, S153, E154, H155, M206, and E223. Alteration of the three acidic residues had major effects on catalysis, changes to S153 and M206 had variable effects on binding and catalysis, and a H155A mutation had only minimal effects on the kinetic parameters. PMID- 16219496 TI - Glucose-dependent insulinotropic polypeptide receptor knockout mice have altered bone turnover. AB - Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone, which is secreted from endocrine cells in the small intestine after meal ingestion. GIP has been shown to affect osteoblastic function in vitro; however, the in vivo effects of GIP on bone remodeling remain unclear. In the present study, we investigated the role of GIP in modulating bone turnover, by evaluating serum markers of bone turnover, bone density, bone morphology, and changes in biomechanical bone strength over time (one to five months) in GIP receptor knockout mice (GIPR-/- mice). The GIPR-/- mice showed a decreased bone size, lower bone mass, altered bone microarchitecture and biomechanical properties, and altered parameters for bone turnover, especially in bone formation. Moreover, the effects of GIP on bone mass were site-specific and compensatory mechanism developed over time and ameliorated the impact of the loss of GIP signaling on bone mass. Further, GIPR-/- mice had earlier age-related changes than wild-type mice in body composition, including bone mass, lean body mass, and fat percentage. In summary, our results indicate that GIP has an anabolic effect on bone mass and bone quality and suggests that GIP may be a hormonal link between nutrient ingestion and utilization. PMID- 16219497 TI - Fate and transport of monoterpenes through soils. Part I. Prediction of temperature dependent soil fate model input-parameters. AB - Monoterpenes are C10H(n)O(n') compounds of natural origin and are potentially environmentally safe substitutes for traditional pesticides. Still, an assessment of their environmental behaviour is required. As a first step in a theoretical study focussing on monoterpenes applied as pesticides to terrestrial environments, soil fate model input-parameters were determined for 20 monoterpenes with widely different structural characteristics. Input-parameters are the water solubility (S(W)), vapour pressure (P), n-octanol-water partition coefficient (K(OW)), atmospheric air and bulk water diffusion coefficients (D(A)air and D(W)water), first order biodegradation rate constants (k), and their temperature dependence. Values for these parameters were estimated or taken from previous experimental work. The quality of the estimations was discussed by focussing on their statistics and by comparison with available experimental data. From these properties, the air-water partition coefficient (K(AW), Henry's Law constant), the interface-water partition coefficient (K(IW)) and the organic matter-water partition coefficient (K(OM)) could be estimated with varying levels of accuracy. In general, little experimental data turned out to be available on biodegradation rate constants and on the temperature dependence of physico chemical parameters. PMID- 16219498 TI - Comparison of a novel passive sampler to standard water-column sampling for organic contaminants associated with wastewater effluents entering a New Jersey stream. AB - Four water samples collected using standard depth and width water-column sampling methodology were compared to an innovative passive, in situ, sampler (the polar organic chemical integrative sampler or POCIS) for the detection of 96 organic wastewater-related contaminants (OWCs) in a stream that receives agricultural, municipal, and industrial wastewaters. Thirty-two OWCs were identified in POCIS extracts whereas 9-24 were identified in individual water-column samples demonstrating the utility of POCIS for identifying contaminants whose occurrence are transient or whose concentrations are below routine analytical detection limits. Overall, 10 OWCs were identified exclusively in the POCIS extracts and only six solely identified in the water-column samples, however, repetitive water samples taken using the standard method during the POCIS deployment period required multiple trips to the sampling site and an increased number of samples to store, process, and analyze. Due to the greater number of OWCs detected in the POCIS extracts as compared to individual water-column samples, the ease of performing a single deployment as compared to collecting and processing multiple water samples, the greater mass of chemical residues sequestered, and the ability to detect chemicals which dissipate quickly, the passive sampling technique offers an efficient and effective alternative for detecting OWCs in our waterways for wastewater contaminants. PMID- 16219499 TI - Differential assimilation of nitrogen dioxide by 70 taxa of roadside trees at an urban pollution level. AB - In order to screen for the best species for mitigating nitrogen dioxide (NO2) by plants at urban levels, we investigated assimilation of nitrogen dioxide by 70 taxa of woody plants that are mostly utilized as roadside trees. They were fumigated with 15N-labeled NO2 at 0.1 microl l(-1) for 8h, and the amount of reduced nitrogen derived from NO2 (in mg Ng(-1) dry weight) in the leaves (designated NO2 assimilation capability hereafter) were determined. Data were analyzed in the comparison with the previously reported ones obtained at 4 microl l(-1) NO2. Among the 70 taxa, the value of NO2 assimilation capability differed by a factor of 122 between the highest (Prunus yedoensis; 0.061) and the lowest (Cryptomeria japonica; 0.0005). Based on the analysis of NO2 assimilation capability values at 0.1 and 4 micro l(-1) NO2, the 70 taxa of woody plants appeared to be classified into four types; those of high NO2 assimilation and high NO2 resistance, those of high NO2 assimilation but low NO2 resistance, those of low NO2 assimilation and low NO2 resistance, and those of low NO2 assimilation but high NO2 resistance. The first, second, third and fourth types include 13, 11, 35 and 11 taxa, respectively. The broad-leaf deciduous trees may have advantages of high biomass and fast growth as compared with woody plants of other habits. Thus, four broad-leaf deciduous species, Robinia pseudo-acacia, Sophora japonica, Populus nigra and Prunus lannesiana, were concluded here to be the best phytoremediators for the urban air. PMID- 16219500 TI - Decrease of atmospheric deposition of heavy metals in an urban area from 1994 to 2002 (Paris, France). AB - Total atmospheric deposition, i.e. both wet and dry ones, was sampled during three different sampling periods between 1994 and 2002. The aim of this study is to determine the temporal variation of the atmospheric deposition fluxes of four heavy metals (Cd, Cu, Pb and Zn) in an urban area (Paris region, France). The global pattern shows a decrease of the fluxes for most of elements during this period. Indeed, the atmospheric deposition fluxes measured in 2001-2002 were lower than those measured during the 1994-1997 period by factors reaching 16, 2.5, 4 and 7.5 at Creteil and 7, 1, 6 and 4.5 at Chatou for Cd, Cu, Pb and Zn, respectively. At the Paris site, the decreasing factors were 2.5 and 3 for Cd and Pb, respectively while Cu and Zn fluxes were nearly similar during the whole studied period. PMID- 16219501 TI - Soil identification and chemometrics for direct determination of nitrate in soils using FTIR-ATR mid-infrared spectroscopy. AB - The use of mid-infrared attenuated total reflectance (ATR) spectroscopy enables direct measurement of nitrate concentration in soil pastes, but strong interfering absorbance bands due to water and soil constituents limit the accuracy of straightforward determination. Accurate subtraction of the water spectrum improves the correlation between nitrate concentration and its nu3 vibration band around 1350 cm(-1). However, this correlation is soil-dependent, due mostly to varying contents of carbonate, whose absorbance band overlaps the nitrate band. In the present work, a two-stage method is developed: First, the soil type is identified by comparing the "fingerprint" region of the spectrum (800-1200 cm(-1)) to a reference spectral library. In the second stage, nitrate concentration is estimated using the spectrum interval that includes the nitrate band, together with the soil type previously identified. Three methods are compared for estimating nitrate concentration: integration of the nitrate absorbance band, cross-correlation with a reference spectrum, and principal component analysis (PCA) followed by a neural network. When using simple band integration, the use of soil specific calibration curves leads to determination errors ranging from 5.5 to 24 mg[N]/kg[dry soil] for the mineral soils tested. The cross-correlation technique leads to similar results. The combination of soil identification with PCA and neural network modeling improves the predictions, especially for soils containing calcium carbonate. Typical prediction errors for light non-calcareous soils are about 4 mg[N]/kg[dry soil], whereas for soils containing calcium carbonate they range from 6 to 20 mg[N]/kg[dry soil], which is less than four percent of the concentration range investigated. PMID- 16219502 TI - The passivation of pyrrhotite by surface coating. AB - The potential of triethylenetetramine (TETA) to inhibit the oxidation of three pyrrhotites, Garson, McCreedy and Po-97 has been studied systematically and confirmed by comparing the release of Fe and SO4(2-) from samples with and without coating treatment. Each sample, original or coated by TETA, was exposed to oxygen, 1 x 10(-3) M FeCl3, and Acidithiobacillus ferrooxidans, respectively, for specific oxidation periods. Both abiotic and biotic oxidation of samples treated by this passivating agent has been reduced significantly in this study. Under the aerobic condition, lower concentrations of ferric, total Fe or SO4(2-) were obtained from the coated samples than those from the uncoated samples. In the presence of 1 x 10(-3) M FeCl3 at 30 degrees C, TETA was able to reduce oxidation rates of Garson, McCreedy and Po-97 by 83%, 79%, and 81% (based on Fe release), respectively. A higher pH, lower Eh, and lower concentrations of total Fe and SO4(2-) were also observed in the biotic oxidation of coated Garson by Acidithiobacillus ferrooxidans. The protection of pyrrhotite surface from oxidant attack by TETA barrier and the alkaline property of this coating agent can be used to interpret the inhibition of oxidation. PMID- 16219503 TI - A comparison of five pesticides adsorption and desorption processes in thirteen contrasting field soils. AB - Batch adsorption and desorption experiments were performed using thirteen agricultural soil samples and five pesticides. Experimental data indicated a gradient in pesticide adsorption on soil: trifluralin >> 2,4-D > isoproturon> atrazine >> bentazone. Atrazine, isoproturon and trifluralin adsorption were correlated to soil organic matter content (r2 = 0.7, 0.82, 0.79 respectively). Conversely, bentazone adsorption was governed by soil pH (r2 = 0.68) while insignificant effect has been shown in the case of 2,4-D. Multiple linear regressions were used to combine relationships between the various soil parameters and the Freundlich adsorption coefficient (K(f)) of each pesticide. Then desorption was assessed since it may reflect some of the interactions involved between the pesticides and the soil components. Adsorbed molecules were released into aqueous solution in the following order: bentazone >> atrazine> isoproturon> 2,4-D >> trifluralin. The occurrence of hysteresis did not allow an accurate interpretation of the pesticide desorption data because of the possible interplay of several processes. PMID- 16219504 TI - Polycyclic musk compounds in higher trophic level aquatic organisms and humans from the United States. AB - Polycyclic musks, 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta[g]-2 benzopyran (HHCB) and 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4 tetrahydronaphthalene (AHTN), are used as fragrance ingredients in numerous consumer products such as cleaning agents and personal care products. Studies have reported the widespread occurrence of these musks in surface waters and fish from western European countries. Nevertheless, little is known about their accumulation in humans and wildlife in the United States. In this study, we measured concentrations of HHCB and AHTN in human adipose fat collected from New York City. Furthermore, tissues from marine mammals, water birds, and fish collected from US waters were analyzed to determine the concentrations of HHCB and AHTN. Concentrations of HHCB and AHTN in human adipose fat samples ranged from 12 to 798 and from <5 to 134 ng/g, on a lipid weight basis, respectively. A significant correlation existed between the concentrations of HHCB and AHTN in human adipose fat. Concentrations of HHCB and AHTN were not positively correlated with age or gender of the donors. HHCB was found in tissues of several wildlife species, but not in the livers of polar bear from the Alaskan Arctic. Among wildlife species analyzed, spinner and bottlenose dolphins collected from Florida coastal waters contained measurable concentrations of HHCB. PMID- 16219505 TI - Copper(II) complexation by humic and fulvic acids from pig slurry and amended and non-amended soils. AB - The effect of the consecutive annual additions of pig slurry at rates of 0 (control), 90 and 150 m3 ha(-1) y(-1) over a 4-year period on the binding affinity for Cu(II) of soil humic acids (HAs) and fulvic acids (FAs) was investigated in a field plot experiment under semiarid conditions. A ligand potentiometric titration method and a single site model were used for determining the Cu(II) complexing capacities and the stability constants of Cu(II) complexes of HAs and FAs isolated from pig slurry and control and amended soils. The HAs complexing capacities and stability constants were larger than those of the corresponding FA fractions. With respect to the control soil HA, pig-slurry HA was characterized by a much smaller binding capacity and stability constant. Amendment with pig slurry decreased the binding affinity of soil HAs. Similar to the corresponding HAs, the binding affinity of pig-slurry FA was much smaller while that of amended-soil FAs were slightly smaller when compared to the control soil FA. The latter effect was, however, more evident with increasing the amount of pig slurry applied to soil per year and the number of years of pig slurry application. PMID- 16219506 TI - Herbicide losses in runoff events from a field with a low slope: role of a vegetative filter strip. AB - Herbicide runoff and the effects of a narrow vegetative filter strip (VFS) were studied on an arable field in the low-lying plains of the Veneto Region (north east Italy). Cultivated plots were compared with and without a 6m wide VFS composed of trees, shrubs and grass. Natural and simulated runoff were monitored during 2000 and 2001. Herbicides applied on the field were: metolachlor (2184 2254 g ha(-1)), terbuthylazine (1000-1127 g ha(-1)) and isoproturon (1000 g ha( 1)). The VFS reduced both runoff depth (10.2-91.2%) and herbicide losses (85.7 97.9%) in the monitored rainfall events. Total herbicide loss with runoff was low (0.69-3.98 g ha(-1) without VFS, less than 0.27 g ha(-1) with VFS), but concentrations were sometimes very high, especially of terbuthylazine and isoproturon during the first events after treatment. In these events there was a high probability of exceeding the ecotoxicological endpoint for algae, but the VFS helped to reduce the potential risk. Two VFS effectiveness mechanisms were identified: (i) dilution, and (ii) a "sponge-like" effect, which temporarily trapped chemicals inside the VFS before releasing them. PMID- 16219507 TI - Lead contamination in tea leaves and non-edaphic factors affecting it. AB - Recent tests have detected high lead (Pb) concentrations in some commercial brands of tea leaves and this finding has raised concerns due to the possible health-related problems associated with Pb poisoning. In present research, we investigated the Pb contamination in tea leaves produced in Zhejiang province in China. Pb concentrations in all tea leaves sampled were below 5 mg/kg, the permissible levels given by Chinese Ministry of Agriculture, indicating that Pb contamination in this province is not excessive. We then investigated the non edaphic factors that may potentially contribute to Pb accumulation in tea leaves. Pb concentration in tea leaves was found to be positively correlated with the industrialization level of a district (R = 0.83, the significant level at P < 0.05), and greater amounts of Pb were washed from the leaves of plants in districts with more industrial activity. This suggests that Pb accumulation in tea leaves could, in part, be attributed to industrial activity through the precipitation of atmospheric Pb. Furthermore greater amounts of Pb were washed from the leaves of plants growing near road than those growing farther away from road. This trend indicates that automobile activity was another likely contributor to Pb accumulation in tea. Pb content of green tea was also affected by the processing of the leaves in the factory. In particular the twisting and water-removal stages caused increases in Pb content in the tea product. This study suggests that non-edaphic factors also contribute to the Pb accumulation in tea. PMID- 16219508 TI - Enhancement of PAH biomineralization rates by cyclodextrins under Fe(III) reducing conditions. AB - Amendment of a soil slurry with low concentrations of a cyclodextrin, hydroxypropyl-beta-cyclodextrin (HPCD), (0.05-0.5 g l(-1)) increased the phenanthrene mineralization rate of a microbial consortium by 25% under Fe(III) reducing conditions. Although a higher concentration (5.0 g l(-1)) resulted in a faster initial rate of mineralization, mineralization ceased after 25 days with maximum mineralization 17% lower than the control (no HPCD). At lower HPCD concentrations, mineralization was still taking place at day 76. Although pH should affect Fe(III) solubility, mineralization rates at pH 6.0 and 8.0 were comparable. Decreasing the temperature reduced the extent and rate of mineralization, but mineralization rates at 10 degrees C were still 60% of that obtained at 30 degrees C. PMID- 16219509 TI - Methods for the isolation and identification of Listeria spp. and Listeria monocytogenes: a review. AB - Listeria monocytogenes is an important food-borne pathogen and is widely tested for in food, environmental and clinical samples. Identification traditionally involved culture methods based on selective enrichment and plating followed by the characterization of Listeria spp. based on colony morphology, sugar fermentation and haemolytic properties. These methods are the gold standard; but they are lengthy and may not be suitable for testing of foods with short shelf lives. As a result more rapid tests were developed based on antibodies (ELISA) or molecular techniques (PCR or DNA hybridization). While these tests possess equal sensitivity, they are rapid and allow testing to be completed within 48 h. More recently, molecular methods were developed that target RNA rather than DNA, such as RT-PCR, real time PCR or nucleic acid based sequence amplification (NASBA). These tests not only provide a measure of cell viability but they can also be used for quantitative analysis. In addition, a variety of tests are available for sub-species characterization, which are particularly useful in epidemiological investigations. Early typing methods differentiated isolates based on phenotypic markers, such as multilocus enzyme electrophoresis, phage typing and serotyping. These phenotypic typing methods are being replaced by molecular tests, which reflect genetic relationships between isolates and are more accurate. These new methods are currently mainly used in research but their considerable potential for routine testing in the future cannot be overlooked. PMID- 16219510 TI - Hydrophobins: the protein-amphiphiles of filamentous fungi. AB - Hydrophobins are surface active proteins produced by filamentous fungi. They have a role in fungal growth as structural components and in the interaction of fungi with their environment. They have, for example, been found to be important for aerial growth, and for the attachment of fungi to solid supports. Hydrophobins also render fungal structures, such as spores, hydrophobic. The biophysical properties of the isolated proteins are remarkable, such as strong adhesion, high surface activity and the formation of various self-assembled structures. The first high resolution three dimensional structure of a hydrophobin, HFBII from Trichoderma reesei, was recently solved. In this review, the properties of hydrophobins are analyzed in light of these new data. Various application possibilities are also discussed. PMID- 16219511 TI - Bio-banking in microbiology: from sample collection to epidemiology, diagnosis and research. AB - Millions of biological samples, including cells of human, animal or bacterial origin, viruses, serum/plasma or DNA/RNA, are stored every year throughout the world for diagnostics and research. The purpose of this review is to summarize the resources necessary to set up a bio-banking facility, the challenges and pitfalls of sample collection, and the most important techniques for separation and storage of samples. Biological samples can be stored for up to 30 years, but specific protocols are required to reduce the damage induced by preservation techniques. Software dedicated to biological banks facilitate sample registration and identification, the cataloguing of sample properties (type of sample/specimen, associated diseases and/or therapeutic protocols, environmental information, etc.), sample tracking, quality assurance and specimen availability. Bio-bank facilities must adopt good laboratory practices and a stringent quality control system and, when required, comply with ethical issues. PMID- 16219512 TI - Mycobacteria in drinking water distribution systems: ecology and significance for human health. AB - In contrast to the notorious pathogens Mycobacterium tuberculosis and M. leprae, the majority of the mycobacterial species described to date are generally not considered as obligate human pathogens. The natural reservoirs of these non primary pathogenic mycobacteria include aquatic and terrestrial environments. Under certain circumstances, e.g., skin lesions, pulmonary or immune dysfunctions and chronic diseases, these environmental mycobacteria (EM) may cause disease. EM such as M. avium, M. kansasii, and M. xenopi have frequently been isolated from drinking water and hospital water distribution systems. Biofilm formation, amoeba associated lifestyle, and resistance to chlorine have been recognized as important factors that contribute to the survival, colonization and persistence of EM in water distribution systems. Although the presence of EM in tap water has been linked to nosocomial infections and pseudo-infections, it remains unclear if these EM provide a health risk for immunocompromised people, in particular AIDS patients. In this regard, control strategies based on maintenance of an effective disinfectant residual and low concentration of nutrients have been proposed to keep EM numbers to a minimum in water distribution systems. PMID- 16219513 TI - Inter-kingdom signaling: deciphering the language of acyl homoserine lactones. AB - Bacteria use small secreted chemicals or peptides as auto-inducers to coordinately regulate gene expression within a population in a process called quorum sensing. Quorum sensing controls several important functions in different bacterial species, including the production of virulence factors and biofilm formation in Pseudomonas aeruginosa and bioluminescence in Vibrio fischeri. Many gram-negative bacterial species use acyl homoserine lactones as auto-inducers that function as ligands for transcriptional regulatory proteins. Several recent reports indicate that bacterial acyl homoserine lactones can also affect gene expression in host cells. Direct signaling also appears to function in the opposite direction as some eukaryotic cell types produce mimics that interact with quorum sensing systems in bacteria. Here, we will describe the evidence to support the existence of bi-directional inter-kingdom signaling via acyl homoserine lactones and eukaryotic mimics and discuss the potential molecular mechanisms that mediate these responses. The functional consequences of inter kingdom signaling will be discussed in relation to both pathogenic and non pathogenic bacterial-host interactions. PMID- 16219514 TI - Polymorphisms of the Nos3 gene and unexplained late intrauterine fetal death. AB - OBJECTIVE: Genetic polymorphisms associated with vascular diseases have been proposed to be involved in the pathogenesis of late unexplained intrauterine fetal death (IUFD). The Nos3 gene is known to regulate vascular tone via the endothelial nitric oxide synthase/nitric oxide pathway. STUDY DESIGN: In a multicenter case-control study, we evaluated two Nos3 polymorphisms (exon 7 Glu298Asp and a 27bp-repeat in intron 4) in 92 women with IUFD and 92 healthy control women. RESULTS: The investigated Nos3 polymorphisms were not associated with the occurrence of IUFD. In the subgroup of pregnancies affected by IUFD, women with at least one mutant allele of the Nos3 intron 4 polymorphism were diagnosed with IUFD at a significantly earlier gestational age (31.8 [standard deviation (S.D.) = 4.9] weeks versus 34.6 [S.D. = 4.8] weeks, p = 0.02) and showed a significantly reduced birth weight (2113 g [S.D. = 1028] versus 1571 g [S.D. = 568], p = 0.03). CONCLUSION: We are the first to report on Nos3 polymorphisms and IUFD. While not being associated with the incidence of IUFD overall, the intron 4 Nos3 polymorphism might modulate the timing of IUFD in affected pregnancies. PMID- 16219515 TI - Expression of CD44 mRNA induced by interleukin-1beta in human cultured uterine cervical fibroblasts. AB - OBJECTIVE: Hyaluronic acid in the cervix plays an important role in the process of cervical ripening. CD44 is a receptor for hyaluronic acid. We investigated whether CD44 is related to cervical ripening by examining the expression of CD44 mRNA in human cultured cervical fibroblasts with or without inflammatory cytokines. STUDY DESIGN: CD44 mRNA expressions derived from cultured human uterine cervical fibroblasts in the presence or absence of interleukin-1beta (IL 1beta) (1-100 pg/ml) or interleukin-8 (IL-8) (1-100 ng/ml) were analyzed by the Northern blot analysis. The human CD44 cDNA probe was hybridized with three species of mRNA. Sizes of the mRNAs were 1.6, 2.2, and 5.0 kilobases (kb). RESULTS: Interleukin-1beta stimulated the expression of CD44 mRNA in a dose dependent manner. Interleukin-8 did not stimulate the expression. CONCLUSIONS: It is concluded from the present study that human cervical CD44 mRNA expression is induced by interleukin-1beta. Accordingly, we deduce that the concentration of hyaluronic acid in the cervical tissue is increased because of the expression of CD44. PMID- 16219516 TI - Caesarean section on demand: influence of personal birth experience and working environment on attitude of German gynaecologists. AB - OBJECTIVE: We performed a survey among German obstetricians and gynecologists in order to evaluate the influence of biographic data, working environment and personal birth experience on the attitude towards Cesarean Section on demand. MATERIAL AND METHODS: All 2106 board-certified gynecologists in Baden-Wurttemberg received an anonymous questionnaire in 2002-2003 concerning attitude towards C section on demand, biographical data, personal birth experience and working environment. Seven hundred and nineteen questionnaires were returned and entered into statistical analysis. RESULTS: General approval of C-section was in 59% of all participants, with huge statistically significant variations according to age, personal birth experience and working field. When asked for their preferred way of delivery for themselves or their partner after a low-risk pregnancy, 90% of the responding gynecologists opted for vaginal delivery. The approval depended statistically significant on parenthood, personal birth experience and working environment. CONCLUSIONS: Biographical data, personal birth experience and working environment influence the attitude towards elective Cesaran section. Although 90% would chose vaginal delivery for themselves or their partner as best medical practice, 59% of the physicians approve of the general opportunity of C section on demand. This shows, that not only best medical practice, but also patient autonomy and forensic aspects seem to play an important role. PMID- 16219517 TI - Perinatal outcome of fetus with isolated congenital second degree atrioventricular block without maternal anti-SSA/Ro-SSB/La antibodies. AB - OBJECTIVE: We determined the perinatal outcomes of fetuses with isolated congenital second degree atrioventricular block detected in utero and born to mothers seronegative for anti-SSA/Ro-SSB/La antibodies. METHODS: Isolated second degree atrioventricular block was defined as second degree atrioventricular block detected in utero without the accompanying structural cardiac anomaly, tachyarrhythmia, non-conducted premature atrial beats or long QT syndrome. We review our own cases and search from Medline using keywords such as atrioventricular block, arrhythmia, bradycardia and congenital to collect cases of congenital isolated second degree atrioventricular block. RESULTS: Two cases were from our institution and five cases from a Medline search; in total seven cases of isolated second degree atrioventricular block without maternal anti SSA/Ro-SSB/La antibodies were analyzed. Six of the seven fetal arrhythmias reverted to sinus rhythm by delivery and did not recur during the follow-up period. The prognosis of the fetus with isolated second degree atrioventricular block without maternal anti-SSA/Ro-SSB/La antibodies is better than that of the fetus with maternal anti-SSA/Ro-SSB/La antibodies or the fetus of congenital long QT syndrome with second degree atrioventricular block detected in utero. CONCLUSION: The fetus with isolated congenital second degree atrioventricular block carries a good prognosis in the absence of maternal anti-SSA/Ro-SSB/La antibodies. PMID- 16219518 TI - Placental localization and expression of the cell death factors BNip3 and Nix in preeclampsia, intrauterine growth retardation and HELLP syndrome. AB - OBJECTIVE: BNip3 and its homologue Nix are pro-apoptotic factors of the Bcl-2 family and are expressed in malignant tumors. In vitro, this expression was shown to be mediated by hypoxia. Recently, it has been shown that placental hypoxia as well as apoptosis are pathogenetic factors for pregnancy-induced hypertensive diseases and intrauterine growth retardation (IUGR). The aim of the study was to analyze placental expression of BNip3 and Nix in pregnancies complicated by preeclampsia, hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome and IUGR. MATERIAL AND METHODS: Placental tissue was sampled from 10 pregnancies each with preeclampsia, HELLP syndrome, IUGR and gestational age matched controls. The placental expression of BNip3/Nix has been investigated with immunohistochemistry by the use of specific human BNip3/Nix antibodies. RESULTS: In cytotrophoblastic cells, the BNip3 expression was strong in the control placentas, but only mediate in the placentas from pregnancies with preeclampsia, IUGR or HELLP syndrome. The intensity of the Nix staining showed a similar pattern. In the syncytiotrophoblast, there was a weak BNip3 staining observable in the control as well as IUGR samples, whereas BNip3 was undetectable in preeclamptic placentas or those with HELLP syndrome. For Nix, only in the preeclampsia a weak staining was detectable, whereas all other probes were negative. CONCLUSIONS: Our study shows for the first time that the pro-apoptotic proteins BNip3 and Nix are expressed in the human placenta. Pregnancies with placental dysfunction and hypertensive pregnancy disorders with different clinical manifestations are characterized by a significantly decreased expression of BNip3 and Nix. These results suggest that the hypothesis of generally increased placental apoptosis in pregnancy-induced hypertensive disorders caused by disturbed trophoblast invasion has to be partly reconsidered. PMID- 16219519 TI - Maternal anemia during pregnancy is an independent risk factor for low birthweight and preterm delivery. AB - OBJECTIVE: The present study was designed to investigate the outcome of pregnancy and delivery in patients with anemia. METHODS: A retrospective population-based study comparing all singleton pregnancies of patients with and without anemia was performed. Deliveries occurred during the years 1988-2002 in the Soroka University Medical Center. Maternal anemia was defined as hemoglobin concentration lower than 10 g/dl during pregnancy. Patients with hemoglobinopathies such as thalassemia were excluded from the analysis. Multiple logistic regression models were performed to control for confounders. RESULTS: During the study period there were 153,396 deliveries, of which 13,204 (8.6%) occurred in patients with anemia. In a multivariable analysis, the following conditions were significantly associated with maternal anemia: placental abruption, placenta previa, labor induction, previous cesarean section (CS), non vertex presentation and Bedouin ethnicity. Higher rates of preterm deliveries (<37 weeks gestation) and low birthweight (<2500 g) were found among patients with anemia as compared to the non-anemic women (10.7% versus 9.0%, p < 0.001 and 10.5% versus 9.4%, p < 0.001; respectively). Higher rates of CS were found among anemic women (20.4% versus 10.3%; p < 0.001). The significant association between anemia and low birthweight persisted after adjusting for gender, ethnicity and gestational age, using a multivariable analysis (OR = 1.1; 95% CI 1.0-1.2, p = 0.02). Two multivariable logistic regression models, with preterm delivery (<37 weeks gestation) and low birthweight (<2500 g) as the outcome variables, were constructed in order to control for possible confounders such as ethnicity, maternal age, placental problems, mode of delivery and non-vertex presentation. Maternal anemia was an independent risk factor for both, preterm delivery (OR = 1.2; 95% CI 1.1-1.2, p < 0.001) and low birthweight (OR = 1.1; 95% CI 1.1-1.2, p = 0.001). CONCLUSION: Maternal anemia influences birthweight and preterm delivery, but in our population, is not associated with adverse perinatal outcome. PMID- 16219520 TI - Predictive power of maternal serum and amniotic fluid CRP and PAPP-A concentrations at the time of genetic amniocentesis for the preterm delivery. AB - OBJECTIVE: To investigate whether maternal serum and amniotic fluid CRP and PAPP A concentrations at the time of genetic amniocentesis are markers of preterm delivery. STUDY DESIGN: One hundred and forty-one pregnant women were included in this prospective study. Amniotic fluid and maternal serum CRP and PAPP-A concentrations were determined by using commercially available kits. Receiver operating characteristic (ROC) analysis was performed to determine the efficacy of maternal serum and amniotic fluid CRP and PAPP-A levels in predicting women with preterm delivery. RESULTS: The prevalence of spontaneous preterm delivery before 37 weeks of gestation was 9.9%. ROC analysis revealed that amniotic fluid CRP level was the only parameter, which had a significant power in the prediction of preterm delivery. The optimum cut-off level was 0.65 mg/L. The sensitivity and specificity were 92.9% and 78.7%, respectively. CONCLUSION: The amniotic fluid CRP level has a high sensitivity and specificity in the prediction of preterm delivery and this may be helpful in predicting preterm delivery during genetic amniocentesis. PMID- 16219521 TI - Obesity and extreme obesity, manifest by ages 20-24 years, continuing through 32 41 years in women, should alert physicians to the diagnostic likelihood of polycystic ovary syndrome as a reversible underlying endocrinopathy. AB - OBJECTIVES: Document obesity-extreme obesity in most 20-24, 25-29, and 32-41 years old women with PCOS as a stimulus for physicians to consider the diagnosis of PCOS, an underlying reversible endocrinopathy. STUDY DESIGN: In matched age groups 20-24, 25-29, and 32-41 years, we compared BMI in 84, 129, and 188 Caucasian women with PCOS versus 956, 815, and 815 women in NHANES I (general population), and 25, 36, and 45 non-pregnant women (community obstetrics practice). RESULTS: At ages 20-24, 25-29, and 32-41 years, mean+/-S.D. BMIs in women with PCOS (35.3+/-7.7, 36.0+/-9.4, 36.7+/-8.2) were much greater than NHANES I (22.8+/-4.6, 23.3+/-5.0, 24.5+/-5.6; p < .0001), and community (26.1+/ 6.8, 26.9+/-6.6, 25.2+/-5.2; p < .0001). Classifying BMI <25 (normal), > or =25 30 (overweight), > or =30-40 (obese), > or =40 (extremely obese), at ages 20-24, 25-29, and 32-41 years: 76, 73, and 78% of PCOS women were obese-extremely obese, versus 7, 11, and 14% of NHANES I, and 20, 28, and 15% of community women. At ages 20-24, 25-29, and 32-41 years, only 10, 12, and 7% PCOS women had BMIs <25, versus 78, 74, and 66% NHANES I, and 48, 47, and 58% of community women. CONCLUSIONS: Obesity-extreme obesity in women, manifest by ages 20-24 years, continuing through 32-41 years, should alert physicians to the likelihood of PCOS, an underlying, heritable, potentially reversible, insulin resistant endocrinopathy that promotes obesity. PMID- 16219522 TI - Interobserver variability in the diagnosis of minimal and mild endometriosis. AB - STUDY OBJECTIVE: To evaluate the observer's influence on the diagnosis and classification of endometriosis according to the rASRM classification. DESIGN: : Prospective analysis. SETTING: University hospital. PATIENTS: Digital videotapes of laparoscopies in three patients with typical endometriotic disorders (rASRM I, II and no endometriosis). INTERVENTION: One hundred and eight gynecologic surgeons were asked to indicate the endometriotic lesions on a prepared surgical sketch and to classify the site according to the rASRM classification. MEASUREMENTS AND MAIN RESULTS: Total number, location and morphology of endometriotic lesions, rASRM classification. The interobserver correlation concerning the number of lesions ranged between 18% (rASRM II) and 30%. (rASRM I). There was marginal correlation regarding the location of endometriotic lesions. Kendall W coefficient ranged from 0.14 (rASRM II) to 0.44 (rASRM I) (p < 0.001). Only 13% (rASRM II) to 22% (rASRM I) of observers used the correct endometriotic classification. None of the participants specified the morphological characteristics of endometriotic lesions according to the rASRM classification. CONCLUSION: Visual assessment of an operative situs with minimal and mild endometriosis is subject to a considerable interindividual variability. One and the same lesion is assessed quite differently by different observers. Histopathological verification seems to be necessary to objectify the diagnosis of endometriosis. PMID- 16219524 TI - Comparison between monofilament and multifilament polypropylene tapes in urinary incontinence. AB - OBJECTIVE: We compare the efficacy and late complications between a polypropylene monofilament tape (TVT) and a polypropylene multifilament tape (IVS) in the surgical implant of sub-urethral, tension free tape for the treatment of stress urinary incontinence (SUI). METHODS: It is a non-randomised, monocentric retrospective study. Two questionnaires were sent to the patients. The first questionnaire determined the efficacy of the tape and the tolerance of the urinary process. The second questionnaire evaluated discomfort and consequences linked to urinary incontinence and associated urinary troubles. RESULTS: The study included 313 female patients and 256 (82%) responded to the questionnaire (137 TVT, 119 IVS). Patient satisfaction was 88.3% for the TVT tape and 73.5% for the IVS tape (p < 0.005). The improvement of the quality of life was also higher for the polypropylene monofilament tapes. "De novo" urge incontinence was more common in the TVT group. However, the rates of dysuria and infection of tapes were more common in the IVS group. CONCLUSION: The type of tape appears to influence efficacy and the emergence of late complications. Our results point at the greater efficacy of the polypropylene monofilament tape. PMID- 16219523 TI - Hypertension is associated with an increased risk for hysterectomy: a Danish cohort study. AB - OBJECTIVE: To assess whether hypertension is a risk factor for hysterectomy, endometrial resection, and myomectomy. STUDY DESIGN: Self-report questionnaires were collected from 81% of 1959 Danish women aged 30 or 40 years selected at random in 1976-1991, in four different cohort studies. Baseline data included standardized information about cardiovascular diseases, hypertension, use of medicine, gynecologic history, social background, and life style factors. Weight, height and blood pressure were measured. The women were followed via central registers to assess the incidence of hysterectomy, endometrial resection, and myomectomy performed for benign diagnoses. Cox regression analyses were used to control for confounding. RESULTS: The average time to follow-up was 15 years, and 135 operations performed for benign diagnoses were identified. Women with a history of hypertension had a double risk of having an operation compared to women without hypertension, independent of confounders. CONCLUSION: Hypertension seems to be a risk factor for hysterectomy performed for benign diagnoses, and thus contributes to women undergoing hysterectomy having an increased risk of cardiovascular diseases. It might be relevant to pay increased attention to the blood pressure in candidates for hysterectomy, as optimal antihypertensive treatment could decrease their otherwise increased risk of cardiovascular disease years after hysterectomy. PMID- 16219525 TI - Sibutramine in pregnancy. PMID- 16219526 TI - Bilateral fallopian tube carcinoma presenting as primary infertility. PMID- 16219527 TI - A useful technique for controlling placental site bleeding for an uncommon type of placentation. PMID- 16219528 TI - Cytokine-based treatment of accidentally irradiated victims and new approaches. AB - A major goal of medical management of acute radiation syndrome following accidental exposures to ionizing radiation (IR) is to mitigate the risks of infection and hemorrhage related to the period of bone marrow aplasia. This can be achieved by stimulating the proliferation and differentiation of residual hematopoietic stem and progenitor cells (HSPC) related to either their intrinsic radioresistance or the heterogeneity of dose distribution. This is the rationale for treatment with hematopoietic growth factors. In fact, apoptosis has recently been shown to play a major role in the death of the continuum of more or less radiosensitive HSPC, soon after irradiation. Therefore, administration of antiapoptotic cytokine combinations such as stem cell factor, Flt-3 ligand, thrombopoietin, and interleukin-3 (4F), may be important for multilineage recovery, particularly when these factors are administered early. Moreover, acute exposure to high doses of IR induces sequential, deleterious effects responsible for a delayed multiple organ dysfunction syndrome. These considerations strongly suggest that therapeutics could include tissue-specific cytokines, such as keratinocyte growth factor, and pleiotropic agents, such as erythropoietin, in addition to hematopoietic growth factors to ensure tissue damage repair and mitigate the inflammatory processes. Noncytokine drugs have also been proposed as an alternative to treat hematopoietic or nonhematopoietic radiation effects. To develop more effective treatments for radiation injuries, basic research is required, particularly to improve understanding of stem cell needs within their environment. In the context of radiological terrorism and radiation accidents, new growth promoting molecules need to be approved and available cytokines stockpiled. PMID- 16219529 TI - Human platelets exhibit infectious-pathogen-binding ligands and participate to inflammation (and more?). PMID- 16219530 TI - A hanging drop culture method to study terminal erythroid differentiation. AB - OBJECTIVE: To design a culture method allowing the quantitative and qualitative analysis of terminal erythroid differentiation. METHODS: Primary erythroid progenitors derived either from mouse tissues or from human umbilical cord blood were differentiated using hanging drop cultures and compared to methylcellulose cultures. Cultured cells were analyzed by FACS to assess differentiation. RESULTS: We describe a practical culture method by adapting the previously described hanging drop culture system to conditions allowing terminal differentiation of primary erythroid progenitors. Using minimal volumes of media and small numbers of cells, we obtained quantitative terminal erythroid differentiation within two days of culture in the case of murine cells and 4 days in the case of human cells. CONCLUSIONS: The established methods for ex vivo culture of primary erythroid progenitors, such as methylcellulose-based burst forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) assays, allow the detection of committed erythroid progenitors but are of limited value to study terminal erythroid differentiation. We show that the application of hanging drop cultures is a practical alternative that, in combination with clonogenic assays, enables a comprehensive assessment of the behavior of primary erythroid cells ex vivo in the context of genetic and drug-induced perturbations. PMID- 16219531 TI - Chelatable cellular copper modulates differentiation and self-renewal of cord blood-derived hematopoietic progenitor cells. AB - OBJECTIVES: We have demonstrated epigenetic modulation of CD34(+) cell differentiation by the high-affinity copper (Cu) chelator tetraethylenepentamine (TEPA). TEPA slowed down the rate of CD34(+) cell differentiation and increased their engraftability in SCID mice. TEPA biological activity was attributed to its effect on cellular Cu levels as (a) treatment with TEPA resulted in reduction of cellular Cu, and (b) excess of Cu reversed TEPA's activity and accelerated differentiation. In the present study we further evaluated the role of cellular Cu in TEPA's biological activity. METHODS: The effects of Cu-chloride, TEPA, TEPA/Cu mixtures at various ratios, and a synthesized, stable, TEPA-Cu complex on short- and long-term cord blood-derived CD34(+) cell cultures as well as on the overall and chelatable cellular Cu were investigated. RESULTS: Addition of TEPA, TEPA/Cu mixtures at up to equimolar concentrations, and the TEPA-Cu complex to CD34(+) cell cultures resulted in inhibition of differentiation and enhancement of long-term self-renewal. Measurement of the overall cellular Cu by atomic absorption spectrophotometry showed 20 to 40% decrease by TEPA while the TEPA-Cu mixture and the TEPA-Cu complex increased cellular Cu by 10- to 20-fold, as did CuCl(2). However, measurement of the cellular pool of labile Cu showed similar reduction (50% from the control) by all the TEPA forms, while CuCl(2) increased it. Thus, inhibition of differentiation and enhancement of self-renewal of CD34(+) cells was correlated with reduction in the cellular chelatable Cu content. CONCLUSION: The results suggest that decreasing of the chelatable Cu pool, rather than overall Cu, is the mechanism that stands behind TEPA's biological activity. PMID- 16219532 TI - CCL23/myeloid progenitor inhibitory factor-1 inhibits production and release of polymorphonuclear leukocytes and monocytes from the bone marrow. AB - OBJECTIVE: CCL23/Myeloid progenitor inhibitory factor-1 is a human CC chemokine with potent in vitro suppressor effects on both human and murine myeloid progenitor cells. This study concerns in vivo inhibitory effect of CCL23 on production of polymorphonuclear leukocytes (PMNs) and monocytes in the bone marrow and their release into the circulation. METHODS: 5'-Bromo-2'-deoxyuridine (BrdU; 100 mg/kg) was used to label dividing PMNs and monocytes in the bone marrow, and BrdU-labeled cells were followed for 10 days in the circulation and identified using immunocytochemistry. Rabbits were given CCL23 (100 mug/kg, n = 5) or saline (control: n = 5) intravenously daily for 3 days before labeling with BrdU. Turnover of PMNs and monocytes in the bone marrow and their transit times through the bone marrow were calculated. RESULTS: CCL23 treatment tended to prolong transit time of PMN (98.4 +/- 4.3 hours vs 111.2 +/- 3.8 hours, control vs CCL23, p = 0.06) through the bone marrow and decreased the size of the bone marrow mitotic pool of PMN (p < 0.01). CCL23 treatment also prolonged the transit time of monocyte (43.4 +/- 3.1 hours vs 54.2 +/- 1.3 hours, control vs CCL23, p < 0.05) through the bone marrow and decreased turnover and pool size of monocytes in the bone marrow (p < 0.05). CONCLUSION: We conclude that CCL23 suppresses PMN and monocyte progenitors, decreases the pool size and slows their turnover in the bone marrow. PMID- 16219533 TI - Proteomic signature of myeloproliferation and neutrophilia: analysis of serum and plasma from healthy subjects given granulocyte colony-stimulating factor. AB - OBJECTIVE: Proteomic analysis could improve our understanding of the mechanisms and consequences of myeloproliferation. Healthy subjects treated with granulocyte colony-stimulating factor (G-CSF) were used as a model of myeloproliferation. METHODS: Levels of 80 soluble factors were measured by enzyme-linked immunosorbent assay before and after 5 days of G-CSF. Both serum and plasma levels were measured to generate a comprehensive profile and determine whether serum or plasma best portrays biological and physiological changes. RESULTS: Comparison of samples collected prior to G-CSF demonstrated that 44 factors differed between serum and plasma. Concentrations of several growth factors and chemokines were greater in serum than in plasma, while the opposite was true for several interleukins. Following G-CSF serum levels of 14 factors and plasma levels of 15 factors changed. Eleven increased in both serum and plasma, including cell adhesion molecules (vascular cell adhesion molecule-1, E-selectin, and L-selectin), matrix metalloproteases (MMP-1, -8, and -13), cytokine receptors (tumor necrosis factor receptor 1 and 2, and interleukin-2 receptor), the acute phase reactant, serum amyloid A, and a growth factor, hepatocyte growth factor. CONCLUSION: Some protein levels differ markedly in serum and plasma. Myeloproliferation is associated with changes in the levels of several proteases, adhesion molecules, and cytokines. PMID- 16219534 TI - High incidence, early onset of histiocytic sarcomas in mice with Hertwig's anemia. AB - OBJECTIVE: Histiocytic sarcoma (HS) is a rare, rapidly disseminated, usually lethal tumor in humans. Treatment specific for HS has not been developed primarily due to deficiencies of appropriate animal models with high incidence/early onset. Mice with Hertwig's anemia (an/an) provide a potential model. METHODS: Here, we compare HS susceptibility in an/an and unaffected control mice maintained on three genetic backgrounds. As a potential therapeutic measure, genetically marked bone marrow is transplanted between high and low susceptibility animals. RESULTS: HS is detected earlier and the overall incidence is 15-fold higher in WBB6F1(F1)-an/an than in F1-+/?, B6-an/an and -+/? mice. Neither WB-an/an nor their normal WB-+/? littermates present with HS. Liver myelopoiesis and aneuploidy coexist with HS but the former is also rampant (33.7% incidence) in HS-free +/? and an/an mice. Marrow transplantation experiments provide evidence that (1) myelopoiesis is associated with HS and (2) early onset/high-incidence HS is blocked by using late-onset F1-+/+ mice, as either donor or recipient. CONCLUSIONS: Homozygosity for an on an F1 genetic background is essential for high-incidence/early-onset HS; myelopoiesis and HS coexist; and therapeutic transplantation may be feasible. PMID- 16219535 TI - A new model of pre-B acute lymphoblastic leukemia chemically induced in rats. AB - OBJECTIVE: Although B acute lymphoblastic leukemia (B-ALL) is the most common leukemia among children, no chemically inducible model of this leukemia has yet been described in vivo. METHODS: Leukemia was chemically induced in male WKAH/Hkm rats by a nitrosourea derivative, N-butylnitrosourea (BNU), an alkylating agent, administered orally 5 days a week for 24 weeks. Development of leukemia was monitored by clinical observation, follow-up of blood parameters, and appearance of blast cells in peripheral blood samples. The phenotype of the leukemia was determined by cytological examination, cytochemical reactions, and by immunophenotyping of bone marrow cells using various markers. The feasibility of leukemia transplantation was investigated. Clonality and karyotype analyses were also performed. RESULTS: We observed the appearance of acute leukemia in 60% of the rats treated with BNU. Of these, 65% developed pre-B-ALL, which was serially transplantable to healthy WKAH/Hkm male rats. Karyotype analysis did not reveal clonal abnormalities. Clonality determined by immunoglobulin gene rearrangement sequencing disclosed that the pre-B-ALL were mostly oligoclonal. CONCLUSION: This new in vivo model of inducible pre-B-ALL might be useful for investigating the effects of co-initiating or promoting agents suspected to be involved in leukemia development, and for disclosing new molecular events leading to leukemogenic processes. PMID- 16219536 TI - Enhanced CML stem cell elimination in vitro by bryostatin priming with imatinib mesylate. AB - OBJECTIVE: In chronic myeloid leukemia (CML), imatinib mesylate (IM; Gleevec, Glivec) induces a G0/G1 cell-cycle block in total CD34(+) cells without causing significant apoptosis. Bryostatin-1 (bryo), a protein kinase C (PKC) modulator, was investigated for its ability to increase IM-mediated apoptosis either through induction of cycling of G0/G1 Ph(+) cells or antagonism of the IM-induced cell cycle block. METHODS: The Ph(+) K562 cell line and primary CD34(+) CML cells were studied for cell-cycle progression (PI staining), proliferation ((3)H thymidine uptake), and survival (dye exclusion). RESULTS: Following 48 hours exposure to IM, on average more than 80% of surviving K562 cells were in G0/G1 as compared to approximately 50% for untreated control cultures (p < 0.001). After accounting for IM-induced cell kill, the absolute number of viable G0/G1 cells was significantly increased, confirming its anti-proliferative effect. However, pretreatment for 24 hours with bryo both increased K562 total cell kill and normalized the percentage of cells recovered in G0/G1, thus reducing their absolute number. For primary CML CD34(+) cells, pretreatment with bryo prior to IM significantly enhanced cell death of both total and, critically, G0/G1 populations. CONCLUSION: These results suggest that carefully scheduled drug combinations that include an agent to antagonize the anti-proliferative effect of IM may prove more efficacious within the Ph(+) stem cell compartment than IM monotherapy. PMID- 16219537 TI - Defective CD3gamma gene transcription is associated with NFATc2 overexpression in the lymphocytic variant of hypereosinophilic syndrome. AB - OBJECTIVE: Determine the molecular defects underlying the CD3(-)CD4(+) T-cell phenotype and persistence of this clonal population in patients with hypereosinophilic syndrome. PATIENTS AND METHODS: Patients in this study suffer from the lymphocytic variant of hypereosinophilic syndrome distinguished by a CD3(-)CD4(+) T-cell clone that overexpresses Th2 cytokines upon activation and thereby provokes the eosinophilia. Interleukin-2-dependent CD3(-)CD4(+) T-cell lines were derived from patient blood at various disease stages and used to investigate the molecular modifications correlated with their abnormal phenotype. RESULTS: We demonstrate that the CD3(-)CD4(+) T cells, characterized by a clonal TCRbeta gene rearrangement, maintained the same immunophenotype over the 6-year period of our study, during which one patient progressed from premalignant disease to CD3(-)CD4(+) T-cell lymphoma. We show that a specific loss of CD3gamma gene transcripts is responsible for the defect in TCR/CD3 surface expression. In addition, the level of NFATc2 binding to NFAT motifs in the CD3gamma gene promoter was greatly increased in the abnormal T cells. Our studies indicate that CD3gamma promoter activity can be positively influenced by NFATc1 plus NF-kappaB p50 and negatively regulated by NFATc2 containing complexes. We show that in patients' CD3(-)CD4(+) T cells, an increase in nuclear NFATc2 occurs in parallel with a decrease in NFATc1 and NF-kappaB gene expression. CONCLUSION: Hypereosinophilic syndrome joins the growing number of pathological conditions where a defect in surface expression and/or function of the TCR/CD3 complex results from altered regulation of CD3gamma gene expression. PMID- 16219538 TI - KHYG-1, a model for the study of enhanced natural killer cell cytotoxicity. AB - OBJECTIVE: To compare the cytotoxicity of KHYG-1 with other natural killer (NK)/NK T-cell lines and identify molecules that may be associated with enhanced cytotoxicity, thereby eventually leading to improved NK cell-mediated cancer immunotherapy. MATERIALS AND METHODS: NK/NK T-cell lines KHYG-1, NK-92, YT, and SNT-8 were compared with a novel flow cytometric cytotoxicity assay under different culture conditions. Transcription, expression, and phosphorylation studies were performed using polymerase chain reaction sequence-specific primers, reverse transcription polymerase chain reaction, immunoblotting, and flow cytometry. RESULTS: KHYG-1 is a highly cytotoxic cell line, exceeding the cytolytic capacity of the other cell lines against K562. KHYG-1 is also highly cytotoxic against the leukemia cell lines EM2, EM3, and HL60. The novel activation receptor NKp44 and its adaptor, DAP12, NKG2D, and constitutively phosphorylated ERK2 may be associated with the enhanced cytotoxicity of KHYG-1. This cell line most likely mediates cytolysis by granzyme M (but not granzymes A and B) together with perforin, which is constitutively fully cleaved to the 60-kD form, in contrast to the other cell lines. CONCLUSION: KHYG-1 is a valuable model for the study of enhanced cytotoxicity by NK cells. In addition to the activation of NKp44, KHYG-1 may induce apoptosis of tumor cells by the newly described granzyme M/perforin pathway. Targeted modifications of effector molecules demonstrated in this model could generate NK cells with even greater killing ability that may be particularly attractive for clinical application. Moreover, our demonstration of greater cytotoxicity of KHYG-1 versus NK-92 cells, already in clinical trials, suggests a direct therapeutic role for KHYG-1. PMID- 16219539 TI - Responses to donor lymphocyte infusion for acute lymphoblastic leukemia may be determined by both qualitative and quantitative limitations of antileukemic T cell responses as observed in an animal model for human leukemia. AB - OBJECTIVE: Donor lymphocyte infusion (DLI) after allogeneic stem cell transplantation is largely unsuccessful in the treatment of acute lymphoblastic leukemia (ALL). To allow identification of the causes of this failure, we established an animal model of DLI in human ALL. METHODS: NOD/scid mice were inoculated with primary human ALL cells. Cells from five different patients were studied. After engraftment, DLI was performed by infusion of human leukocyte antigen (HLA)-identical donor T cells or HLA-disparate donor T cells. RESULTS: DLI resulted in expansion of activated, leukemia-reactive T cells in all donor patient combinations. After 40 days of expansion, T cells abruptly declined in numbers and displayed loss of cytotoxicity. At this moment, remissions were observed in three of five donor-patient combinations. In animals engrafted with the two unresponsive ALL, remissions could be achieved when HLA-disparate DLI was given. CONCLUSION: Our results indicate that the inefficacy of DLI in ALL may be due to the limitation of the proliferative capacity of ALL-reactive T cells and that the antileukemic efficacy during the limited time span of proliferation depends on the antigenic disparity between the donor and the patient. The model can be used to study whether alternative strategies may result in more sustained antileukemic responses after DLI in ALL. PMID- 16219540 TI - Efficient in vitro megakaryocyte maturation using cytokine cocktails optimized by statistical experimental design. AB - OBJECTIVE: A multi-step statistical strategy was applied to quantify individual and interactive effects of cytokines on megakaryopoiesis and to determine the concentration of the selected cytokines that optimize ex vivo megakaryocyte (MK) expansion, maturation, and platelet production in stromal- and serum-free conditions. MATERIALS AND METHODS: Immature MK were first generated from human CD34(+)-enriched cord blood cells cultured for 7 days in conditions favoring MK commitment. Then, the effect of different combinations of cytokines at various concentrations on MK differentiation and platelet production was tested on the day-7 MK. RESULTS: A large-scale screening of 13 cytokines in the presence of thrombopoietin (TPO) using Placket-Burman designs (PBD) was initially performed to identify stimulators of MK maturation. Afterwards, a statistical analysis of the two-level factorial designs revealed that in the presence of TPO, MK maturation was significantly stimulated by stem cell factor (SCF), interleukin (IL)-6, and IL-9, whereas Flt-3 ligand (FL) had a positive effect only on the expansion of MK progenitors. In contrast, erythropoietin (EPO) and IL-8 were inhibitors of MK maturation. A response surface methodology was then used to optimize the concentrations of the selected cytokines (TPO, SCF, IL-6, and IL-9) and defined a new cytokine cocktail that maximized MK expansion and maturation. Importantly, the increased MK output was accompanied by a very high MK purity ( approximately 90%). Another optimum was also found at a higher SCF concentration, which further improved MK expansion and maturation, but reduced MK purity. CONCLUSION: These statistical methods provide an efficient tool to analyze complex systems of cytokines and to develop promising ex vivo MK culture systems for clinical applications. PMID- 16219541 TI - MMP-2 is required for bone marrow stromal cell support of pro-B-cell chemotaxis. AB - OBJECTIVE: We have previously demonstrated that bone marrow stromal cells (BMSCs) exposed to etoposide (VP-16) have reduced support of CXCR4(+) cell chemotaxis and diminished stromal cell derived factor-1 (CXCL12) in the supernatants. Based on the identification of CXCL12 as a matrix metalloproteinase-2 (MMP-2) substrate, we investigated potential dysregulation of MMP-2 expression or activity in chemotherapy-treated BMSCs. METHODS: BMSCs exposed to VP-16 were evaluated for MMP-2 expression by gelatin zymography, ELISA, and western blot. Chemotaxis assays were completed to evaluate pro-B cell chemotaxis toward either MMP-2(-/-) BMSCs or BMSCs exposed to MMP-2 inhibitors. RESULTS: BMSC exposure to VP-16 resulted in an immediate, transient, increase in MMP-2, followed by reduced MMP-2 protein expression. MMP-2 reduction correlated with diminished CXCL12 protein and reduced support of pro-B cell chemotaxis. BMSCs derived from MMP-2 knockout mice had less chemotactic support of CXCR4(+) cells than wild-type controls. Inhibition of BMSC MMP-2 activity by OA-Hy also reduced chemotactic support and CXCL12 protein detected in BMSC supernatants. VP-16-induced reduction of BMSC support of hematopoietic cell migration was restored by supplementing cultures with recombinant MMP-2 protein. CONCLUSIONS: These data suggest that MMP-2 is sensitive to chemotherapy-induced stress, and may regulate BMSC support of pro-B cell chemotaxis. Increased MMP-2 expression during the acute phase of chemotherapy exposure potentially inactivates CXCL12. Subsequently, chronic exposure to chemotherapy, with the associated downregulation of MMP-2, interrupts CXCL12 release from the extracellular matrix, also blunting BMSC support of pro-B cell migration. PMID- 16219542 TI - Adipocyte differentiation in Sod2(-/-) and Sod2(+/+) murine bone marrow stromal cells is associated with low antioxidant pools. AB - OBJECTIVE: Adipocytogenesis in bone marrow stromal cells (BMSCs) from manganese superoxide dismutase-deficient (Sod2(-/-)) and wild-type (Sod2(+/+)) mice and the effect of antioxidant pool size were determined. METHODS: BMSCs from Sod2(-/-) or Sod2(+/+) mice were cultured with and without adipocytogenic supplements including: 10 mug/mL insulin, 1 muM dexamethasone, and 100 muM indomethacin. Oil Red-O-positive cells and reverse-transcriptase polymerase chain reaction measurement of peroxisome proliferator-activated receptor-gamma (PPARgamma) and lipoprotein lipase (LPL) were measured. Antioxidant glutathione levels (GSH) and glutathione peroxidase activity (GPX) were determined. RESULTS: Sod2(-/-) cells demonstrated constitutive adipocytogenesis in basal medium and generated 34% more adipocytes in adipocytogenic media. Growth of cells in the free radical scavenger antioxidant, amifostine (WR2721; 4 mM) decreased numbers of adipocytes in Sod2(-/ ) BMSCs in both basal (38.0%, p = 0.037) and adipocytogenic (37.5%, p = 0.021) media and reduced to undetectable the levels of expression of PPARgamma and LPL. In contrast, Sod2(+/+) cells showed no detectable constitutive adipocytogenesis but formed adipocytes in adipocytogenic medium, with a decrease (43.7%, p = 0.001) by addition of WR2721. In basal conditions, Sod2(-/-) cells had lower GSH (78.6%; p = 0.0089) and GPX (52.7%; p < 0.001) levels than did Sod2(+/+) cells, which were increased in either medium by WR2721 treatment of Sod2(-/-) or Sod2(+/+) cells (all p < 0.001). Differentiation of BMSCs to adipocytes was inversely correlated with the level of GSH (r = -0.9427, p = 0.0167). Sod2(-/-) long-term bone marrow cultures had decreased hematopoiesis compared to those from Sod2(+/-) or Sod2(+/+) mice. CONCLUSION: The cellular redox pathway has a role in adipocyte differentiation of cells of the hematopoietic microenvironment. PMID- 16219543 TI - Age-associated skewing of X-inactivation ratios of blood cells in normal females: a candidate-gene analysis approach. AB - X-inactivation is a random process that occurs in females early during embryogenesis. Females are mosaics with an equal proportion of cells with the paternal (Xp) or maternal X-chromosome (Xm) in the active state. However, close to 40% of healthy females aged more than 60 y.o. present a significant skewing of X-inactivation ratios (Xp:Xm >3 :1). The exact etiology of this age-associated skewing (AAS) in blood cells is unknown. We hypothesized that AAS is due to hemizygous cell selection caused by allelic variants in hematopoiesis or cell survival genes. To test this hypothesis, we recruited 700 unrelated healthy females of French Canadian ancestry aged more than 60. We determined X inactivation ratio at the HUMARA locus. We genotyped 81 different SNPs, using TaqMan technology, in 15 different candidate genes with known role in hematopoiesis, cell cycle, or X-inactivation. Extensive statistical analyses were conducted and demonstrated that none of the 15 candidate genes investigated contribute significantly to AAS. PMID- 16219544 TI - Congenital amegakaryocytic thrombocytopenia in three siblings: molecular analysis of atypical clinical presentation. AB - OBJECTIVE: An 11-year-old girl, presenting with fatigue and bruising, was found to be profoundly pancytopenic. Bone marrow exam and clinical evaluation were consistent with aplastic anemia. Family members were studied as potential stem cell donors, revealing that both younger siblings displayed significant thrombocytopenia, whereas both parents had normal blood counts. We evaluated this pedigree to understand the unusually late presentation of congenital amegakaryocytic thrombocytopenia (CAMT). MATERIALS AND METHODS: The coding region and the intron/exon junctions of MPL were sequenced from each family member. Vectors representing each of the mutations were constructed and tested for the ability to support growth of Baf3/Mpl(mutant) cells. RESULTS: All three siblings had elevated thrombopoietin levels. Analysis of genomic DNA demonstrated that each parent had mutations/polymorphisms in a single MPL allele and that each child was a compound heterozygote, having inherited both abnormal alleles. The maternal allele encoded a mutation of the donor splice-junction at the exon 3/intron-3 boundary. A mini-gene construct encoding normal vs mutant versions of the intron-3 donor-site demonstrated that physiologic splicing was significantly reduced in the mutant construct. CONCLUSIONS: Mutations that incompletely eliminate Mpl expression/function may result in delayed diagnosis of CAMT and confusion with aplastic anemia. PMID- 16219545 TI - Intrinsic regulation of the interactions between the SH3 domain of p85 subunit of phosphatidylinositol-3 kinase and the protein network of BCR/ABL oncogenic tyrosine kinase. AB - OBJECTIVE: BCR/ABL fusion tyrosine kinase is responsible for the initiation and maintenance of the Philadelphia chromosome-positive chronic myelogenous leukemia (CML) and a cohort of acute lymphocytic leukemias. We show that a signaling protein, phosphatidylinositol-3 kinase (PI-3k), is essential for growth of CML cells, but not of normal hematopoietic cells, and that p85alpha subunit of PI-3k co-immunoprecipitates with BCR/ABL. Therefore, we made an attempt to better characterize p85alpha-BCR/ABL interactions. MATERIALS AND METHODS: The mutants of p85alpha-SH3 domain were generated by in vitro site-directed mutagenesis system. Protein lysates were obtained from p210BCR/ABL-transformed murine 32Dcl3 myeloid cells, and in vitro transcription/translation was used to produce BCR/ABL protein. Pull-down and Western analyses were performed to detect the interaction between BCR/ABL and p85alpha-SH3. BCR/ABL-transformed 32Dcl3 cells were infected with internal ribosome entry site-green fluorescent protein retroviral construct encoding p85alpha-SH3 mutants to assess their biological effects. RESULTS: We show here that the SH3 domain of p85alpha (p85alpha-SH3) pulls down the p210BCR/ABL kinase from hematopoietic cell lysates. The interaction between p85alpha-SH3 and BCR/ABL may be intermediated by proteins such as c-Cbl, Shc, Grb2, and/or Gab2. Mutations in the p85alpha-SH3 region responsible for proline rich motif binding either abrogate or enhance these interactions. These mutants exert a modest inhibitory effect on growth factor-independent proliferation of BCR/ABL-positive 32Dcl3 cells. CONCLUSIONS: Based on this information we speculate on the capability of p85alpha-SH3 to interact with the protein network of BCR/ABL oncoprotein. PMID- 16219546 TI - Dynamic changes in cellular and microenvironmental composition can be controlled to elicit in vitro human hematopoietic stem cell expansion. AB - OBJECTIVE: The absence of effective strategies for the ex vivo expansion of human hematopoietic stem cells (HSCs) limits the development of many cell-based therapies. Prior attempts to stimulate HSC expansion have focused on media supplementation using cytokines and growth factors. In these cultures, cellular and microenvironmental compositions change with time. In this study, the impact of controlling these dynamic changes on HSC output is determined. MATERIALS AND METHODS: Cord blood-derived lin(-) cells were cultured for 8 days in serum-free medium supplemented with stem cell factor, Flt3 ligand, and thrombopoietin. Functional, phenotypic, and molecular (gene and protein) analyses were used to characterize dynamic changes in cellular and microenvironmental composition. The effects of these changes and the mechanism behind their effects on HSC expansion were assessed using a selection/media exchange-based global culture manipulation (GCM) technique. RESULTS: We show that the direct secretion of negative regulators by culture-generated lin(+) cells, and the indirect stimulation of cells to secrete negative regulators by culture-conditioned media, limits in vitro HSC generation. The GCM strategy was able to abrogate these effects to produce elevated numbers of LTC-ICs (14.6-fold relative to input), migrating rapid NOD/SCID repopulating cells (12.1-fold), and long-term NOD/SCID repopulating cells (5.2-fold). CONCLUSIONS: Cellular and microenvironmental changes that occur during all in vitro HSC cultures can significantly affect HSC output through the direct or indirect secretion of negative regulators. This study provides insight into the mechanisms regulating HSC fate in vitro and describes a novel methodology to regulate overall in vitro microenvironmental dynamics to enable the generation of clinically relevant numbers of HSCs. PMID- 16219547 TI - Reduced-intensity conditioning regimen preserves thymic function in the early period after hematopoietic stem cell transplantation. AB - OBJECTIVE: To compare T-cell reconstitution in two groups of patients submitted to allogeneic stem cell transplantation (SCT): those receiving reduced-intensity conditioning (RIC, n = 24) and those receiving myeloablative conditioning (MA, n = 27). METHODS: Fifty-one consecutive patients undergoing SCT were evaluated. Serial assessments of lymphocyte subsets and T cell receptor excision circles (TRECs) levels were performed using multiparametric flow cytometry and real-time PCR, respectively. RESULTS: During the first 6 months posttransplant, total and naive CD4(+) T cell counts were higher after RIC-SCT than after MA-SCT (total CD4(+): p = 0.04, p = 0.08, and p = 0.058; naive CD4(+): p = 0.14, p = 0.05, and p = 0.01 at 1, 3, and 6 months, respectively). In both groups of patients, TRECs levels were low or undetectable in the first 3 months after SCT and progressively increased during the study. However, a higher proportion of patients with detectable levels of TRECs was observed in RIC-SCT at 1 and 3 months and more patients in this group reached normal levels of TRECs at 6 months post-SCT. In the multivariate analysis, including factors such as type of donor (sibling vs unrelated), dose of CD34(+) cells infused with the graft, patient age, and graft vs-host disease (GVHD), the most important factor influencing TRECs recovery in the early period after SCT was the type of conditioning regimen. CONCLUSIONS: In this study, the pattern of immune reconstitution after RIC-SCT was different from that of MA-SCT and was characterized by higher posttransplant naive CD4(+) T cell counts and TRECs levels in the early period after transplant. PMID- 16219548 TI - Enhanced engraftment of umbilical cord blood-derived stem cells in NOD/SCID mice by cotransplantation of a second unrelated cord blood unit. AB - OBJECTIVE: Umbilical cord blood (UCB) is considered as an attractive alternative source of hematopoietic stem cells for allogeneic stem cell transplantations in patients who lack human leukocyte antigen (HLA)-matched donors. However, the low cell dose adversely affects hematopoietic recovery and therefore limits application of UCB transplantation in adults. Transplantation of multiple UCB units could be a strategy to overcome cell dose limitations. MATERIALS AND METHODS: To investigate the effect of double cord transplantation, nonobese diabetic/severe combined immunodeficient mice were transplanted with human hematopoietic progenitor cells (CD34(+)) derived from two UCB units with HLA disparity. Human cell engraftment and donor origin was determined by flow cytometry. RESULTS: Double CB transplantation resulted in increased engraftment levels in the bone marrow and peripheral blood in comparison with recipients of a single unit. Because this effect could be due to the higher cell dose (2.10(5) vs 1.10(5) cells), double CB transplantation was compared with single units containing equal cell numbers (2.10(5)). In some cases, engraftment levels in recipients of single units containing 2.10(5) cells were significantly higher than after transplantation of 1.10(5) cells. These engraftment levels were similar to those observed after double CB transplantation. Chimerism analysis indicated that increased engraftment in recipients of two units was predominantly derived from one unit, whereas in other cases the contribution of the two units was similar. CONCLUSION: These results indicate that engraftment may be enhanced by addition of a second unrelated CB that might be attributed to a cell dose effect or due to a graft-facilitating effect. PMID- 16219549 TI - Influence of telomere length on short-term recovery after allogeneic stem cell transplantation. AB - OBJECTIVE: Telomeres shorten in somatic cells during aging and states of increased turnover, including hematopoietic stem cell transplantation. Fast hematopoietic recovery is critical for the patients' course after hematopoietic stem cell transplantation. It is unknown whether telomere length in hematopoietic stem cells (HSCs) predicts short-term hematopoietic recovery. METHODS: We quantified telomere length by flow fluorescence in situ hybridization analysis in HSCs and granulocytes of healthy stem cell donors and monitored time to peripheral blood cell recovery in transplanted hosts. Furthermore, we measured in vitro repopulation potency of HSCs by assaying for colony-forming units granulocyte-macrophage (CFU-GM). RESULTS: Telomere length in HSC shortens continuously in vivo and is comparable to telomere length in granulocytes from the same individual. Numbers of in vitro formed CFU-GM per HSC show an inverse relationship to age and telomere length. However, telomere length in HSCs was not correlated with short-term recovery after HSC transplantation. CONCLUSION: These findings suggest that healthy stem cell donors have sufficient telomere length reserve to repopulate a myeloablatively treated host, despite continuous aging of HSCs in vivo and decreased repopulation ability of HSCs from older donors in vitro. PMID- 16219550 TI - Thalassemia major: the iron irony. PMID- 16219551 TI - Synthetic peptide analogs derived from bcr/abl fusion proteins induce heteroclitic human T cell responses. PMID- 16219552 TI - Immune surveillance of leukemia? PMID- 16219553 TI - Pegylated interferon for the treatment of high-risk essential thrombocythemia. PMID- 16219554 TI - Outcome after allogeneic bone marrow transplantation in cytogenetic high-risk myelodysplastic syndromes and acute myeloid leukemia. PMID- 16219555 TI - Evolving strategies for the management of high-risk adult acute lymphoblastic leukemia. PMID- 16219556 TI - Bendamustine in relapsed or refractory chronic lymphocytic leukemia. PMID- 16219557 TI - Minimal residual disease detection in myeloma: no more molecular remissions? PMID- 16219558 TI - Chimerism analysis after allogeneic stem cell transplantation. PMID- 16219559 TI - Anti-leukemic effect of graft-versus-host disease on bone marrow and extramedullary disease following allografting. PMID- 16219560 TI - CD25 expression on donor CD8+ T cells and graft-versus-host disease. PMID- 16219561 TI - Hepatic veno-occlusive disease following stem cell transplantation: younger patients are not sheltered from this complication. PMID- 16219562 TI - T time. PMID- 16219563 TI - Gene transfer of HA-1 specificity. PMID- 16219564 TI - Combined therapy with deferiprone and desferrioxamine in thalassemia major. PMID- 16219565 TI - Evaluation and monitoring of response to therapy in multiple myeloma. PMID- 16219566 TI - Combined therapy with deferiprone and desferrioxamine in thalassemia major. AB - BACKGROUND AND OBJECTIVES: Effective and convenient iron chelation remains one of the main targets of clinical management of thalassemia major. The combined treatment with desferrioxamine and deferiprone could have an increased chelation efficacy and sometimes allow drug doses and toxicity to be reduced and the number of days of desferrioxamine infusion to be decreased, improving compliance and quality of life. DESIGN AND METHODS: We used combined therapy with desferrioxamine and deferiprone to treat 79 patients with severe iron overload (serum ferritin higher than 3000 ng/mL) who had low compliance with subcutaneous desferrioxamine. RESULTS: Total therapy exposure was 201 patient-years. Three patients developed agranulocytosis and seven mild neutropenia. Other adverse effects were nausea, vomiting, abdominal pain, increased concentrations of liver transaminases and joint pain. The efficacy of combined therapy was evaluated in 64 patients treated for at least 12 months. Ferritin decreased from 5243+/-2345 to 3439+/-2446 ng/mL, p<0.001). Mean urinary iron excretion during combined therapy was double that with desferrioxamine or deferiprone monotherapy. In 20 patients receiving heart therapy at baseline, left ventricular ejection fraction increased from 48.6+/-9% to 57+/-6% (p=0.0001) over 12 to 57 months, without modifying the cardiac treatment. INTERPRETATION AND CONCLUSIONS: Continuous deferiprone treatment with intermittent administration of subcutaneous desferrioxamine is a practical and effective procedure to decrease severe iron overload in patients with thalassemia major. This study also shows that the combined therapy is associated with an improvement in heart function. PMID- 16219567 TI - Circulating bcr-abl-specific CD8+ T cells in chronic myeloid leukemia patients and healthy subjects. AB - BACKGROUND AND OBJECTIVES: The fusion oncoprotein bcr-abl that characterizes chronic myeloid leukemia (CML) is a leukemia-specific antigen, which may be immunogenic in vivo. KQSSKALQR and GFKQSSKAL, peptide sequences spanning the b3a2 bcr-abl junction, have affinity for HLA-A3 and HLA-B8, respectively, and we have shown the presence of KQSSKALQR on the surface of CML cells. We analyzed the existence of bcr-abl-specific T cells in vivo and correlated their presence to contemporary disease burden. DESIGN AND METHODS: We investigated circulating CD8+ T lymphocytes directed against the bcr-abl junction, using fluorochrome-labeled tetramers of HLA-A3 with KQSSKALQR and of HLA-B8 with GFKQSSKAL, and flow cytometry analysis. Using chromium-release assays and interferon-g ELISPOT assays, we also studied the functionality of these expanded T cells. RESULTS: Eight of 12 b3a2+ HLA-A3+ and/or HLA-B8+ CML patients studied serially on at least three occasions had bcr-abl junction-specific CD8+ T cells. Specific T cells were more likely to be found in patients with a low leukemic burden (p=0.03). Three of 18 HLA-A3+ and/or HLA-B8+ healthy donors had bcr-abl junction specific T cells, though these were not detected in any of 13 subjects who were HLA-A3- and HLA-B8-. Bcr-abl-specific T cells were expandable in vitro in three of seven healthy donors and five of seven CML patients. INTERPRETATION AND CONCLUSIONS: Bcr-abl-specific T cells are detectable in CML patients, and might contribute to leukemic control. The occurrence of specific CD8+ T cells in some healthy donors might represent an immune response to occult BCR-ABL rearrangements. PMID- 16219568 TI - Synthetic peptide analogs derived from bcr/abl fusion proteins and the induction of heteroclitic human T-cell responses. AB - BACKGROUND AND OBJECTIVES: Chronic myelogenous leukemia (CML) presents a unique opportunity to develop therapeutic strategies using vaccination against a truly tumor-specific antigen that is also the oncogenic protein required for neoplasia. We have shown in phase I and II trials that a tumor-specific, bcr-abl-derived peptide vaccine can be safely administered to patients with chronic phase CML and can elicit a reliable specific CD4 immune response. However, variable CD8 responses and no HLA A0201-restricted responses were found. One strategy to circumvent this poor immunogenicity is to design synthetic immunogenic analog peptides that cross-react with the native peptides (a heteroclitic response). The aim of this study was to design such peptides. DESIGN AND METHODS: By using computer prediction analysis. We designed a number of synthetic peptides derived from the junctional sequences of CML (p210/b3a2 or p210/b2a2) in which single and double amino acid substitutions were introduced at key HLA A0201 binding positions. The binding of these peptides was tested by a thermostabilization assay using a T2 cell line. RESULTS: We found three peptides that predicted good binding to HLA A0201 molecules and stabilized MHC class I A0201 molecules on the surface of T2 cell lines. These peptides were screened for eliciting HLA restricted, peptide-specific cytotoxic T lymphocyte responses using CD3+ T cells from several A0201 donors and CML patients. The CD8+ cytotoxic T lymphocytes lines were assessed by either interferon-g ELISPOT or a chromium release assay using pulsed, HLA-matched leukemic cell lines. The analog peptides generated larger immune responses (increased CD8 T-cell precursor frequency) than did the native peptides. Importantly, CD8+ T cells stimulated with the new synthetic peptides cross-reacted with the native bcr-abl peptides. INTERPRETATION AND CONCLUSIONS: In conclusion, analog CML fusion peptides with increased immunogenicity and heteroclitic properties can be synthesized and may be useful in vaccination strategies. PMID- 16219569 TI - Pegylated interferon for the treatment of high risk essential thrombocythemia: results of a phase II study. AB - BACKGROUND AND OBJECTIVES: Patients with high-risk essential thrombocythemia require cytoreductive therapy in order to normalize the elevated platelet counts. We evaluated the efficacy and toxicity of pegylated interferon in high-risk essential thrombocythemia in a phase II trial. DESIGN AND METHODS: Thirty-six patients with high-risk essential thrombocythemia (median age 54 years; range, 24 72 years) were studied. The dose of pegylated interferon was initially 50 mg per week and could be escalated up to 150 mg per week. RESULTS: During the first three months platelet counts decreased significantly from a median baseline count of 895x10(9)/L (range: 383-1779) to a median count of 485x10(9)/L (range: 211 1283; p<=0.001). A complete response was defined as platelet counts < 450x10(9)/L. The complete response rate was 39%, 47%, 58% and 67% at 3, 6, 9 and 12 months of treatment, respectively. There were 25%, 11%, 8% and 0% poor responders, defined as patients with platelet counts > 600x10(9)/L, at 3, 6, 9 and 12 months of treatment, respectively. After a median time of 23 months (range 3-39 months) 23 of 36 patients (64%) are still receiving pegylated interferon. In ten patients (28%) treatment was stopped due to grade 1 to 2 toxicity, classified according to the WHO standard toxicity scale. One patient, who responded partially to pegylated interferon (platelet count 542x10(9)/L), had a cerebral stroke after 23 months of treatment. INTERPRETATION AND CONCLUSIONS: In high-risk essential thrombocythemia sustained treatment with pegylated interferon is effective and safe in reducing platelet counts with a toxicity comparable to that of conventional interferon. PMID- 16219570 TI - Allogeneic stem cell transplantation for patients with acute myeloid leukemia or myelodysplastic syndrome who have chromosome 5 and/or 7 abnormalities. AB - BACKGROUND AND OBJECTIVES: Chromosome 5 and/or 7 abnormalities are cytogenetic findings indicative of a poor prognosis in patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). The only potential cure for such patients is allogeneic stem cell transplantation (SCT). As data on allogeneic SCT in this context are limited we did a retrospective study of allogeneic SCT in patients with AML or MDS who had chromosome 5 and/or 7 abnormalities. DESIGN AND METHODS: This was a retrospective study of 65 patients (16 children, 49 adults) with AML (n=33) or MDS (n=32) who had chromosome 5 and/or 7 abnormalities and who underwent allogeneic SCT in six Dutch Centers between 1983 and 2001. Data on all these patients are recorded in the Netherlands Stem Cell Transplant Registry (Typhon). RESULTS: The 3-year overall survival rate among all patients was 25%. Patients below the age of 40 years had significantly fewer relapses (40%) and better survival (38%) than those above the age of 40 (86% and 8%, respectively). Relapses were less frequent in recipients of unrelated grafts than in those whose grafts were from HLA-identical siblings (30% versus 69%). The development of acute graft-versus-host disease (GVHD) grades II-IV was independently associated with significantly higher transplant-related mortality (TRM). Patients with either chromosome 5 or chromosome 7 abnormalities had a significantly better survival than patients with both chromosome 5 and 7 abnormalities. These patients with poor-risk chromosome 5 and/or 7 abnormalities were compared with a group of patients with a secondary AML/MDS and normal cytogenetics and were found to have significantly more relapses and significantly worse survival but a similar TRM. INTERPRETATION AND CONCLUSIONS: We conclude that patients with AML or MDS with chromosome 5 and/or 7 abnormalities do rather poorly after allogeneic SCT, mainly because of the very high relapse rate. Nevertheless, this is the only approach that can cure some of these patients. PMID- 16219571 TI - Comparison of intensive chemotherapy, allogeneic or autologous stem cell transplantation as post-remission treatment for adult patients with high-risk acute lymphoblastic leukemia. Results of the PETHEMA ALL-93 trial. AB - BACKGROUND AND OBJECTIVES: The optimal post-remission therapy for adults with high-risk acute lymphoblastic leukemia (ALL) is not well established. This multicenter randomized trial by the Spanish PETHEMA Group was addressed to compare three options of post-remission therapy in adults with high-risk ALL: chemotherapy, allogeneic stem cell transplantation (SCT) and autologous SCT. DESIGN AND METHODS: A total of 222 valid high-risk ALL patients entered the trial. All received a standard five-drug/five-week induction course. Patients in complete remission with an HLA-identical family donor were assigned to allogeneic SCT (n=84) and the remaining were randomized to autologous SCT (n=50) or to delayed intensification followed by maintenance chemotherapy up to 2 years in complete remission (n=48). RESULTS: Overall, 183 patients achieved complete remission (82%). With a median follow-up of 70 months, the median disease-free survival and overall survival were 17 and 23 months, respectively. The 5-year disease-free survival and overall survival were 35% (95% CI, 30%-41%) and 34% (95% CI, 28%-39%), respectively. Patients allocated to the chemotherapy, allogeneic and autologous SCT were comparable in the main pre-treatment ALL characteristics and the rate of response to therapy. Intention-to-treat analysis showed no differences between patients according to whether they had or did not have a donor in disease-free survival (39%, 95% CI 30-48% vs. 33%, 95% CI 23-41%) and overall survival (44%, 95% CI 35-52% vs. 35%, 95% CI 25-44%), as well as for autologous SCT vs. chemotherapy comparisons (disease-free survival: 40%, 95% CI 28-52% vs. 51%, 95% CI 37-67%; overall survival: 43%, 95% CI 29-58% vs. 52%, 95% CI 39-65%). No differences were observed when the analysis was made on the basis of the treatment actually performed. INTERPRETATION AND CONCLUSIONS: This study failed to prove that, when a family donor is available, allogeneic SCT produces a better outcome than autologous SCT or chemotherapy in adults with high-risk ALL. PMID- 16219572 TI - Efficacy of bendamustine in patients with relapsed or refractory chronic lymphocytic leukemia: results of a phase I/II study of the German CLL Study Group. AB - BACKGROUND AND OBJECTIVES: Although bendamustine has been used for more than 30 years in the treatment of lymphoma, little is known about the optimal dosing schedule in relapsed or refractory B-cell chronic lymphocytic leukemia (CLL). Various dose and treatment schedules have been used empirically, and several phase II studies have shown impressive efficacy. To determine the maximal tolerated dose, dose-limiting toxicity and the optimal therapeutic dose of bendamustine for further phase III clinical trials the GCLLSG designed a phase I/II study for pre-treated CLL patients. DESIGN AND METHODS: Sixteen patients (median age 67 years) with relapsed or refractory CLL were enrolled. All patients had been pre-treated with a median of three different regimens. Bendamustine was given at a starting dose of 100 mg/m2 on day 1 and 2, repeated every 3-4 weeks. RESULTS: Major toxicities were leukocytopenia (CTC grade 3+4) in 8/16 and infections (CTC grade 3+4) in 7/16 patients. Six patients had dose-limiting toxicity which led to dose de-escalation from 100 to 70 mg/m2 in three patients. The maximum tolerated dose was 70 mg/m2. According to NCI-WG criteria, 9/16 patients (56%) responded to therapy, seven to doses or=18 years) known during 1997-1998 to 15 Dutch hospitals were selected for inclusion. Each patient was followed for 3 years, and resource use associated with each of the treatments, including watchful waiting, was recorded. The hospital perspective was adopted. Unit costs were based on 2003 price levels. RESULTS: Two hundred patients were included of whom 75% underwent one or more treatments during the 3-year data collection period [25% were not treated because of a watchful waiting strategy (10%) or complete remission (15%)]. Allogeneic and autologous stem cell transplantations were the most expensive treatments, with a mean (median) per patient cost of 45,326 euro(44,237; n=7) and 18,866 euro (16,532; n=9), respectively (up to discharge only). Intravenous fludarabine cost 10,651 euro (9,995; n=33), rituximab (10,628 euro; 10,124; n=7), and CHOP 7,547 euro (5,833; n=42). Classical FL treatments were found to be the least expensive treatments used with an estimated cost for cylophosphamide, vincristine and prednisone of 5,268 euro (2,644; n=58), for radiotherapy of 4,218 euro (4,313; n=52), and for chlorambucil of 2,476 euro (1,098; n=53). INTERPRETATION AND CONCLUSIONS: This study presents information on resource use and costs associated with the most commonly prescribed FL treatments. In addition to differences in effectiveness, commonly used treatments vary considerably in terms of resource use and overall cost. This information is of value for resource planning, given the high costs of new treatment modalities. PMID- 16219579 TI - Adult and cord blood T cells can acquire HA-1 specificity through HA-1 T-cell receptor gene transfer. AB - BACKGROUND AND OBJECTIVES: Minor histocompatibility antigen (mHag)-specific graft versus-leukemia reactivities are observed following unselected donor lymphocyte infusion for the treatment of relapse after HLA-matched mHag-mismatched stem cell transplantation (SCT). Adoptive transfer of donor-derived ex vivo-generated HA-1 specific oligoclonal T cells or HA-1 peptide patient vaccination are currently being explored as curative tools for stem cell based immunotherapy of hematologic malignancies. Another treatment modality to eradicate residual leukemic cells after SCT is the transfer of the HA-1 hematopoietic-specific T-cell receptor (TCR) into cells from the stem cell donor. This strategy would be particularly useful in case of relapse after cord blood transplantation (CBT) and is explored in this study. DESIGN AND METHODS: HLA-A2(neg) adult peripheral blood and cord blood mononuclear cells were transduced with the genes encoding the HA-1 alpha and beta TCR chains derived from established HA-1 specific cytotoxic T lymphocyte clones. RESULTS: The T cells transduced with HA-1 TCR alpha beta showed consistent marker gene expression, but low staining with HLA-A2/HA-1 tetrameric complexes. They did, however, show hematopoietic-restricted cytolytic activity against HLA-A2(pos)/HA-1(pos) target cells, including leukemic cells. INTERPRETATION AND CONCLUSIONS: The low level of HA-1-specific tetramer staining of HA-1 TCR alpha beta transduced T cells may be caused by hybrid TCR formation of the transferred TCRalpha and beta chains with endogenous TCR alpha and beta chains. This may cause unwanted alloreactivity and requires attention. The HA-1 TCR alpha beta transduced T cells show that the HA-1 TCR can be functionally transferred into donor mononuclear cells, which can be exploited in immunotherapeutic settings of SCT and CBT for hematologic malignancies. PMID- 16219581 TI - Inhibition of the mammalian target of rapamycin and the induction of cell cycle arrest in mantle cell lymphoma cells. AB - Mammalian target of rapamycin (mTOR) inhibitors represent a new class of potential anticancer agents. The mTOR inhibitor, rapamycin, inhibited proliferation in three mantle cell lymphoma (MCL) cell lines and reduced cyclin D3 expression while cyclin D1 levels remained unchanged. This finding was confirmed in cells from a MCL patient. PMID- 16219582 TI - The efficacy of alemtuzumab for refractory chronic lymphocytic leukemia in relation to cytogenetic abnormalities of p53. AB - We reviewed the efficacy of alemtuzumab in the treatment of 28 patients with refractory chronic lymphocytic leukemia (CLL) in whom p53 status was known. Overall responses of 53.6% (complete responses 17.9%) were attained with no significant difference between patients with (50%) or without (55%) p53 deletion (p=0.214). We confirm the efficacy of alemtuzumab in refractory CLL irrespective of p53 deletions, and advocate its introduction earlier in disease course. PMID- 16219583 TI - Serum syndecan-1 in patients with newly diagnosed monoclonal proteinemia. AB - Serum syndecan-1 was investigated in 189 patients with newly diagnosed monoclonal proteinemia [the diagnoses were multiple myeloma (66), monoclonal gammopathies of undetermined significance (MGUS; n=54), provisional MGUS (no bone marrow examination performed; n=69)] and 36 controls. Syndecan-1 levels ranged widely between all diagnostic categories and were of limited discriminatory value (sensitivity 68%, specificity 78%) in patients with newly diagnosed monoclonal proteinemia. PMID- 16219584 TI - Depression and associated factors among women within their first postnatal year in Erzurum province in eastern Turkey. AB - OBJECTIVES: The aims of the study were to identify associated factors with depression, to estimate prevalence of depression among women in the postnatal first year in Eastern Turkey, and to compare the results with those obtained from western societies. METHODS: We randomly selected 728 women in their postnatal first year. The data were collected in 2003 by using a structured questionnaire and the Edinburgh Postnatal Depression Scale. RESULTS: The percentage of the women who had scores > or = 13 was 34.6%. Having an unemployed husband, premenstrual symptoms, lack of husband's support, any stressful life event during the pregnancy, infant with a health problem, a temperamentally difficult child, and history of psychiatric condition were found to be associated factors with depression in women in this region. CONCLUSION: The percentage of women with high scores in Eastern Turkey was higher than the reported prevalence in many western countries, and significant relations were identified between depression and social factors. PMID- 16219585 TI - Temporal changes in anthropometric measurements of idealized females and young women in general. AB - This study examined the temporal anthropometric changes in idealized female body images in the media (i.e., Playboy magazine Playmates of the Year, Miss America Pageant winners, and fashion models) and young women in general across eight decades. Overall, all anthropometric measures differed significantly over time. BMI for all women in the idealized groups tended to decline significantly over time, while the BMI for YoungWomen increased significantly. Models tended to have the smallest bust and hips, Playmates the largest bust, and young women in general the largest waist and hips. The general trend for all groups was to move from a less curvaceous body shape in the early part of the twentieth century to a more curvaceous shape at mid-century and returning to a less curvaceous shape at the end of the century. Idealized women have a body size unlike that of Young Women and the chasm between the media- defined ideal and reality is continuing to diverge. PMID- 16219586 TI - Women on top: the relative influence of wives and husbands on contraceptive use in KwaZulu-Natal. AB - OBJECTIVES: The purpose of this paper is to examine the relative influence of husband and wife on contraceptive practice. METHODS: A cross-sectional survey was conducted among adult men and women in KwaZulu-Natal, South Africa. A matched file for 238 married or cohabiting couples was created. RESULTS: Knowledge of methods of contraception was virtually universal and attitudes to contraception were favorable both in men and women. A substantial proportion of men and women reported using a method of contraception. The wife's desire to stop childbearing was the most powerful predictor of contraceptive use among couples, after adjustment for possible confounders. The husband's approval (or not) of family planning and his preference for future childbearing were not significantly related to contraceptive use. CONCLUSION: Contraceptive use within marital and cohabiting unions is high and the wife's fertility preference was found to be a key determinant of use. This conclusion challenges conventional wisdom that men are the dominant decision-makers in fertility and family planning decisions. PMID- 16219587 TI - Sources of social support as predictors of health, psychological well-being and life satisfaction among Dutch male and female dual-earners. AB - We examined whether gender differences in health, psychological well-being, and life satisfaction, can be explained by effects of work-related and nonwork related sources of social support. The sample consisted of 459 men and women from dual earner families. Men report better health and psychological well-being than women, whereas women report higher life satisfaction than men. Contrary to our expectations, women receive more social support from colleagues than men, while men and women equally receive support from their supervisor. As for the nonwork related sources of social support, men receive more social support from their spouse, while women receive more social support from relatives and friends. No gender differences exist in the effects of social support. Although men and women differ with respect to the social support they receive from different sources, these differences cannot explain gender differences in health, psychological well being and life satisfaction. PMID- 16219588 TI - HIV, STD, and hepatitis risk to primary female partners of men being released from prison. AB - Incarcerated men in the US are at increased risk for HIV, STDs and hepatitis, and many men leaving prison have unprotected sex with a primary female partner immediately following release from prison. This paper addresses risk to the primary female partners of men being released from prison (N = 106) by examining the prevalence of men's concurrent unprotected sex with other partners or needle sharing prior to and following release from prison (concurrent risk). Rates of concurrent risk were 46% prior to incarceration, 18% one month post release, and 24% three months post release. Multivariate analysis showed concurrent risk was significantly associated with having a female partner who had one or more HIV/STD risk factors and having a history of injection drug use. Findings demonstrate need for prevention programs for incarcerated men and their female partners. PMID- 16219589 TI - Women's decision making about whether or not to use breast cancer chemoprevention. AB - BACKGROUND: Minimal research is available that examines how women at high risk for breast cancer make choices about chemoprevention. We examined how women decide whether to use chemoprevention within the Study of Tamoxifen and Raloxifene (STAR), a non-placebo arm prevention trial. DESIGN: We used in-depth, semi-structured interviews and qualitative analysis. We interviewed 28 participants who chose to take chemoprevention and 23 STAR eligible women who declined participation and chemoprevention. RESULTS: Most participants perceived themselves at heightened risk for and anxious about developing breast cancer and believed taking either trial medication was a reasonable way to reduce risk. More than half of non-participants did not perceive themselves at heightened risk for and felt little anxiety about breast cancer. More than half of non-participants felt at greater risk for developing other diseases. CONCLUSIONS: While breast cancer risk can be calculated with a fair degree of accuracy, its impact on women's choices about chemoprevention is likely best assessed in tandem with women's feelings about developing breast cancer, whether within the context of a clinical trial or not. PMID- 16219590 TI - Experiences of women in a minimal contact pedometer-based intervention: a qualitative study. AB - BACKGROUND: Minimal contact pedometer-based interventions are gaining in popularity as a cost-effective method of promoting physical activity. The experiences of women in these interventions, however, have not been adequately studied. PURPOSE: This study used focus groups to explore women's experiences in a minimal contact pedometer-based intervention. METHODS: Participants were women aged 31 to 51 years who completed a 6-week minimal contact pedometer-based intervention. Before the intervention, all participants were inactive or irregularly active. For the intervention, participants wore pedometers, completed weekly logs of daily steps taken, created physical activity goals, and received weekly emails. Email messages contained strategies for increasing physical activity and served as reminders to wear the pedometer and submit the step logs. After the intervention, participants were invited to attend focus groups to discuss their experiences in the intervention. A generic qualitative research approach was used to analyze the data. RESULTS: Wearing the pedometer helped participants set goals and motivated them to increase their physical activity. Submitting step logs made them accountable. They wanted more innovative tips in the emails for increasing physical activity. DISCUSSION: Our results suggest that the effectiveness of minimal contact interventions may be enhanced by including pedometers, step logs, and email reminders. PMID- 16219591 TI - Medicare reform--a wolf in sheep's clothing. AB - This article examines the Medicare Prescription Drug, Improvement and Modernization Act of 2003 as a case study of the political process and examines the elements of the legislation, the process of passage, and implications of some its most controversial components. PMID- 16219593 TI - Assuring the quality of long-term care insurance benefits through care management: the California partnership for long-term care. AB - Despite recent improvements in long-term care insurance (LTCI) policies, concerns have been raised regarding just how well LTCI benefits actually meet elderly consumers' health and financial needs. In this case study, we examined the quality assurance (QA) provisions in a state-sponsored LTCI program, the California Partnership for Long-Term Care (CPLTC). CPLTC invests the primary responsibility for QA with care management networks, which assure quality services through care monitoring, quarterly service record reviews, and annual documentation of care manager clinical competence. Study findings suggest a number of limitations in existing QA policies and procedures, which can undermine the ability of care managers and other third parties to identify and rectify potential unmet needs among LTCI policyholders. These findings, while based on an intensive analysis of QA provisions in a particular, state-sponsored LTCI program, are likely to have implications for other LTCI programs and policies, most of which have less well-developed QA provisions. PMID- 16219592 TI - Provider dilemmas with relocation in assisted living: philosophy vs. practice. AB - Support for increasing tenant frailty in assisted living (AL) is a complex and challenging issue. Philosophically, older persons, families, providers, and policymakers want to avoid unnecessary relocation. However, there is considerable opportunity for inconsistency in practice. We examined provider perspectives related to implementing aging-in-place, using information obtained from an evaluation of a three-year AL demonstration project in Illinois. We found differences in approaches to explicit move-out criteria, the actual application of move-out criteria, and provider willingness to develop flexible service plans. These findings raise questions for consideration in all states confronted with implementing the philosophy and intent of AL. PMID- 16219594 TI - Long-term care insurance in Japan. AB - Among all the industrialized countries, Japan has the fastest rate of population aging and the highest life expectancy at birth. It is projected that the proportion of elderly people will reach 35.7% in 2050. In this demographic environment, Japan launched a social insurance program for long-term care for the elderly in 2000. What were the forces that led Japan to establish a long-term care program for elderly people? What are the provisions for financing, benefits, and service delivery? What aspects of policymaking in developing such a program are unique to Japan?. PMID- 16219595 TI - Exploring attitudes toward older workers among Australian employers: an empirical study. AB - Research suggests that negative stereotyping is at the heart of age discrimination in not hiring older workers. As the aging of the population creates pressures to maintain older adults in the work force, it is important to gain knowledge of such stereotyping. A random sample of 128 hiring decision makers across Australian industries responded to a questionnaire that was specifically developed to assess attitudes toward older workers. Results showed systematic negative stereotyping: Respondents indicated that they were unlikely to hire older workers. The likelihood of hiring significantly correlated with employers' attitudes. Specific attributes for which older workers were seen as inferior compared to younger workers included trainability, adaptability, creativity, and interest in new technology. The findings could inform policymakers and be utilized to develop interventions aimed to reduce hiring discrimination. PMID- 16219597 TI - Understanding the racial and ethnic differences in caregiving arrangements. AB - In this study, the relative importance of family/household structure, socioeconomic status, and culture is examined, and their connection to racial and ethnic variations in caregiving networks is explored. Each of these three domains is seen as contributing to the race/ethnic variations in caregiving arrangements. PMID- 16219598 TI - Who's afraid of growing old? Gay and lesbian perceptions of aging. AB - This article is a study of how gay men and lesbian individuals perceive the aging process. The findings indicate that gay men have more negative views of how gay society views growing older and how they view their own growing older than do lesbian respondents. Gay men were also found to be more ageist, have a greater fear of negative evaluation by others, and give more importance to their own physical attractiveness. Implications for social work practice with elder homosexuals are addressed. PMID- 16219599 TI - Men who work at age 70 or older. AB - The federal policy on older workers has shifted from the encouragement of early withdrawal from the labor force to the encouragement of continuous participation in the labor force. In this light, it is instructive to investigate the backgrounds of elderly people who work at age 70 or older. This article presents the findings of a study, using data from the 1993 Asset and Health Dynamics of the Oldest Old Study, that investigated the effects of health, economic conditions (net worth, employer-provided pensions, and supplemental medical insurance coverage), education, and spouse's work status on the probability of working among men aged 70 or older. The study addressed the probability of working, the probability of working fulltime and of working part-time, and the probability of being self-employed and of being employed by others. Implications for policy are discussed. PMID- 16219600 TI - Factors associated with the positive well-being of grandparents caring for their grandchildren. AB - Prior research on grandparent caregivers has focused on stress, depression, and other negative outcomes. This paper describes research on factors that are associated with grandparents' positive well-being. In particular, it investigated the extent to which the perception of grandparental stress and grandparents' resources are associated with grandparents' well-being, after controlling for sociodemographic and health factors. A sample of 129 grandparents had individual interviews. Using a hierarchical block multiple regression analysis, the authors found that a low perception of stress related to caring for grandchildren and resources were responsible for a high level of wellbeing. The findings of this study suggest that social workers can best help grandparent caregivers by lowering their perception of stress and enhancing their informal supports and community resources. This can be incorporated into supportive, strengths-based individual or family counseling. PMID- 16219601 TI - Social-economic context of parent care: explaining Chinese caregivers' psychological and emotional well-being. AB - This paper explores Chinese familial caregivers' depression and subjective burdens among the current caregivers. Data were collected in 1997-1999 with 110 caregivers who were then taking care of physically dependent parents or parents in-law. Regression analyses were conducted to assess the factors that influence caregivers depression and subjective burden. Findings suggest that family and individual economic conditions are related to caregivers' depression. Caregivers' involvement in caregiving tasks is associated with caregivers' subjective burden. Poorer self-perceived health is related to higher report of caregivers' depression. The author argues that poorer health and unemployment at the time of drastic social and economic changes in China may have created psychological and emotional anxieties and depression for caregivers. The lack of financial and social stability may have deleterious effect for current caregivers in the future when they grow older and have to be cared for by their one-child generation children. PMID- 16219602 TI - The nursing home crisis: a consumer study of Texas nursing home care. AB - This article discusses the findings of a consumer-oriented survey to ascertain the perceptions of Texas nursing home residents, their families, and nursing home administrators about what factors they believe contribute to quality of care. The findings from previous studies and established practice standards are then used to further delineate the practice roles and functions of social workers in nursing facilities (NFs) and to inform social work curriculum. PMID- 16219605 TI - Pharmacotherapists need to know non-drug therapies. PMID- 16219606 TI - Physician perspectives on a pilot prescription monitoring program. AB - In 2003, during implementation of a pilot electronic prescription monitoring program in southwest Virginia, a survey was mailed to 672 physicians to learn about their knowledge of and attitudes toward the program and its impact on their opioid prescribing behaviors. A total of 275 surveys were returned yielding a response rate of 41%. Less than one-half had previously heard about the prescription monitoring program. Nearly 60% believed their prescribing behaviors were being monitored more closely as a result of the program; of these, 23% reported that this had a negative impact on their ability to help patients manage their pain. Sixty-eight percent reported that the prescription monitoring program was useful for monitoring patients' prescription histories and decreasing doctor shopping; however, only 11% had requested information from the prescription monitoring program database, primarily due to access barriers. Recommendations include education to increase physician awareness of and utilization of the program and to address their concerns about scrutiny of practice and collection of relevant data that examines the impact of the program on diversion, abuse, and quality of patient care for persons in pain. PMID- 16219607 TI - Neurolytic celiac plexus block: a better alternative to opioid treatment in upper abdominal malignancies: an Indian experience. AB - The majority of patients with advanced upper abdominal malignancies suffer from moderate to severe pain due to unavailability of morphine in developing world. This study was undertaken to evaluate the role of neurolytic celiac plexus block on pain and quality of life in this patient subpopulation. One hundred consecutive patients receiving opioids for their pain relief were divided in two groups. Group I (control) patients received oral morphine & NSAIDS and group II (study) patients underwent neurolytic celiac plexus block (NCPB) to compare their effects on pain relief, morphine consumption, quality of life (QOL), Karnofsky and performance scores up to one month. NCPB provided statistically significant better pain relief and reduced morphine consumption at one month (P = 0.000). Superior Karnofsky and performance scores also favored NCPB group (P = 0.000); however the difference in overall QOL was not statistically significant (P = 0.24). Patients in oral morphine group had more side effects (94% vs. 58%) as compared to NCPB (P = 0.000). NCPB is an effective tool to reduce opioid requirement and the drug-related adverse effects. It is a rewarding technique, especially when morphine availability and its easy accessibility to the deserving patient is poor. PMID- 16219608 TI - The effect of music therapy on postoperative pain, heart rate, systolic blood pressures and analgesic use following nasal surgery. AB - The prevalence of unrelieved postoperative pain is high and may lead to adverse effects including prolonged hospitalization and delayed recovery. Distraction may be an effective pain-relieving strategy, and can be implemented by several means including affective imaging, games, and possibly music. The aim of this study was to explore the effect of music therapy on postoperative pain. Fifty-seven patients (24 females, 33 males; mean +/- SD age 39.9 +/- 14.35 years [range 15 to 69 years] were matched for age and sex and then nonselectively assigned to either an experimental (n = 27) or a control (n = 30) group. Music was played intermittently to members of the experimental group during the first 24 hour postoperative period. Pain intensity was measured using the Pain Verbal Rating Scales (VRS). Significant decreases in pain intensity over time were found in the experimental group compared to the control group (p < 0.0001). In addition, the experimental group had a lower systolic blood pressure and heart rate, and took fewer oral analgesics for pain. These findings suggest that music therapy is an effective nonpharmacologic approach for postoperative pain management. PMID- 16219609 TI - Aprepitant (EMEND): the role of substance P in nausea and vomiting. AB - Aprepitant (EMEND) is the first commercially available drug from a new class of agents, the Substance P/neurokinin NK-1 receptor antagonists. Aprepitant is indicated for prevention of acute and delayed chemotherapy-induced nausea and vomiting (CINV) associated with highly emetogenic chemotherapy in adults. Its ability to antagonize the effects of Substance P has lead to greater understanding of the pathophysiology of nausea and vomiting. Its broad range of activity against a wide variety of central and peripheral emetogenic stimuli make it potentially useful in non-chemotherapy related nausea and vomiting. PMID- 16219610 TI - Prostate cancer presenting with normal serum PSA levels and boney metastases treated with zoledronic acid. AB - Prostate specific antigen (PSA) is currently the most widely used tumor marker for diagnosing and monitoring prostate cancer. Serum PSA level greater than 10 ng/mL is considered a negative predictor for the presence of bone metastases and routine use of bone scans is therefore avoided. This report describes two cases of patients presenting with normal PSA levels (0-4 ng/ mL) and positive bone scans. The value of routine use of bone scintigraphy in the follow up of these cases is therefore questioned here. Moreover the addition of bisphosphonates to the androgen deprivation treatment, achieved not only pain relief and improvement in aspects of quality of life, but also provided radiographic evidence of management of metastatic bone disease. PMID- 16219611 TI - Sound-alike drug names produce risk of medication interchange errors. AB - Due to the risk of medications errors from inadvertent interchange of drugs for which names look alike or sound alike, the Joint Commission on Accreditation of Healthcare Organizations now requires that accredited health care organizations develop and maintain programs to minimize these risks. This report lists drug names that can lead to such errors. PMID- 16219614 TI - Epidural steriods. AB - The use of steroids administered into the epidural space to manage low back pain is described in a manner that a clinician might use to explain this intervention to a patient or care-giver. PMID- 16219612 TI - Neuroleptics in terminal restlessness. PMID- 16219615 TI - Bisphosphonates for metastatic bone pain. AB - Bisphosphonates are a class of medications used increasingly to help manage pain due to bony metastases in cancer patients. Newer bisphosphonates used in this way are reviewed and the pharmacokinetic are compared. PMID- 16219616 TI - Evidence-based pain management and palliative care in issue one for 2005 of The Cochrane Library. AB - The Cochrane Library of Systematic Reviews is published quarterly. The Library now contains 2249 complete reviews, 1539 protocols of reviews and 5009 one page summaries of systematic reviews published in the general medical literature. In addition there are citations of 435,786 randomised controlled trials, 19 methodology reviews and 6370 cited papers in the Cochrane methodology register. The health technology assessment database contains 4516 citations. In this edition of the Library issued in January 2005, there were seven new reviews of which six have potential relevance for practitioners in pain and palliative medicine. Last year, the format of the citation has changed with the move to publication of The Cochrane Library by John Wiley & Sons. Citations will probably change again in the future as currently there is no indication of the date when a review was first published. PMID- 16219617 TI - World Wide Web resources for cancer support groups. AB - World Wide Web (internet) resources that pertain to cancer support groups, especially pain and symptom control, are listed and reviewed. How to start using these services; services for patients, families, and friends; disease-specific websites; and other types of electronic information repositories are discussed. PMID- 16219618 TI - National Institutes of Health State-of-the-Science Conference statement. Improving End-of-Life Care. December 6-8, 2004. AB - The draft conference statement is presented in its entirety. This report addresses five important questions, i.e. What defines the transition to end of life; What outcome variables are important indicators of the quality of the end of-life experience for the dying person and for the surviving loved ones; What patient, family, and health care system factors are associated with improved or worsened outcomes; What processes and interventions are associated with improved or worsened outcomes; What are the future research directions for improving end of-life care? The report includes conclusions, a list of panel members, speakers, the planning committee and conference sponsors. PMID- 16219619 TI - Pain as a human right: the 2004 Global Day Against Pain. AB - On October 11, 2004, the International Association for the Study of Pain (IASP), the World Health Organization (WHO), and the European Federation of IASP Chapters (EFIC) held a Global Day against Pain campaign to promote pain relief as a human right. International leaders in pain management and public health participated in the full day program that took place in Geneva Switzerland and was simultaneously broadcast around the world through the Word Wide Web. The declaration and statements that were released and announced, and selected contents of the program are summarized in this report. PMID- 16219622 TI - Pharmacokinetics and efficacy of ifosfamide or trofosfamide in patients with intraocular lymphoma. AB - BACKGROUND: The prognosis of intraocular lymphoma (IOL) is poor, and the optimal treatment has not yet been defined. The study assesses ifosfamide (IFO) and trofosfamide (TRO) for treating IOL. PATIENTS AND METHODS: We prospectively evaluated the efficacy and aqueous penetration of intravenous IFO, oral TRO and their active 4-hydroxy (4-OH) metabolites in 10 patients with IOL. Doses varied from 1500 to 2000 mg/m2/day on days 1-3 for IFO and from 150 to 400 mg/day (continuous or intermittent administration) for TRO. Four patients had newly diagnosed disease, and six had relapsed after pretreatment. RESULTS: All patients responded to first treatment with IFO or TRO, and both of two patients responded to re-treatment with IFO on ocular relapse. Progression-free survival from the first treatment with IFO or TRO was > or = 6-18 months. In six of six patients, 4 OH metabolites were detected in the aqueous humor at a concentration of 0.32-1.56 microM immediately after IFO infusion with an aqueous/serum ratio of 0.19-0.54. 4 OH metabolites could be detected in one of three patients at a concentration of 7.2 microM 3-16 h after ingestion of TRO. CONCLUSIONS: IFO and TRO are active in IOL. IOL patients evidence aqueous penetration of 4-OH metabolites after intravenous administration of IFO. PMID- 16219623 TI - Phase II study of preoperative oxaliplatin, capecitabine and external beam radiotherapy in patients with rectal cancer: the RadiOxCape study. AB - BACKGROUND: Preoperative radiotherapy has been shown to decrease the local recurrence rate of patients with locally advanced rectal cancer. Capecitabine and oxaliplatin are both active anticancer agents in the treatment of patients with advanced colorectal cancer and have radiosensitizing properties. Therefore, these drugs would be expected to improve effectiveness of preoperative radiotherapy in terms of local control and prevention of distant metastases. PATIENTS AND METHODS: Forty patients with rectal cancer (T3-T4 and/or N+) received radiotherapy (1.8 Gy, 5 days a week over 5 weeks, total dose 45 Gy, 3D conformational technique) in combination with intravenous oxaliplatin 50 mg/m2 once weekly for 5 weeks and oral capecitabine 825 mg/m2 twice daily on each day of radiation. Surgery was performed 6-8 weeks after completion of radiotherapy. The main end points were safety and efficacy as assessed by the pathological complete response (pCR). RESULTS: The most frequent grade 3/4 adverse event was diarrhea, occurring in 30% of patients. pCR was found in five (14%) patients. According to Dworak's classification, good regression was found in six (18%) additional patients. CONCLUSIONS: Combination of preoperative radiotherapy with capecitabine and oxaliplatin is feasible for downstaging rectal cancer. PMID- 16219624 TI - Differential expression of biomarkers in lung adenocarcinoma: a comparative study between smokers and never-smokers. AB - BACKGROUND: Non-small-cell lung cancer arising in never-smokers is usually of adenocarcinoma subtype. The oncogenic pathway of such tumors is poorly understood. To better define the biological characteristics of these tumors, we have compared the expression of a panel of epidermal growth factor receptor (EGFR)-related biomarkers in lung adenocarcinomas from smokers versus those in never-smokers. PATIENTS AND METHODS: Using immunohistochemical analysis, we retrospectively analyzed EGFR, pAKT, PTEN, Ki-67, p27 and hTERT expression in specimens from 190 patients with completely resected lung adenocarcinomas (43 never-smokers and 147 smokers). These analyses were performed on tissue microarrays. RESULTS: EGFR expression was higher in tumors from smokers (P < 0.01), while pAKT was overexpressed mainly in tumors from never-smokers (P = 0.01). As expected, the tumors from smokers presented a higher expression of Ki 67 and a more frequent loss of expression of p27 (P < 0.01). In a multivariate model, two biological factors (p27 and Ki-67) and two clinical factors (age and sex) showed independent significant correlation with never-smoking status. CONCLUSIONS: Lung adenocarcinomas in never-smokers have a very distinct immunohistochemical expression profile of EGFR-related biomarkers as compared with lung adenocarcinomas in smokers. High levels of EGFR and Ki-67 are observed in smokers, while never-smokers are characterized by high levels of pAKT and p27. PMID- 16219625 TI - Selective inhibition of HER2 inhibits AKT signal transduction and prolongs disease-free survival in a micrometastasis model of ovarian carcinoma. AB - Although first-line chemotherapy induces complete clinical remission in many cases of epithelial ovarian cancer, relapse usually occurs 18-28 months from diagnosis owing to micrometastases. The present study aimed to evaluate the effect of trastuzumab on disease-free and overall survival in a specially designed murine model of ovarian cancer (OVCAR-3), which mimicked the natural history of human micrometastatic disease. Trastuzumab can cure the mice if started soon after induction chemotherapy. It can modestly inhibit the proliferation through mitogen-activated protein kinase signal transduction and clearly inhibit AKT phosphorylation, which is involved in survival pathway. As OVCAR-3 cell lines show no HER2 amplification or overexpression, these results warrant further studies to assess the efficacy of trastuzumab in the early stage of relapse in cancer models other than those overexpressing HER2. PMID- 16219626 TI - Deletions in the polyAlanine-containing transcription factor FOXL2 lead to intranuclear aggregation. AB - Mutations of FOXL2, a gene encoding a forkhead transcription factor, have been shown to cause the blepharophimosis-ptosis-epicanthus inversus syndrome. This genetic disorder is characterized by eyelid and craniofacial abnormalities associated or not with premature ovarian failure. We have previously shown that mutant FOXL2 with an expanded polyAlanine (polyAla) tract forms large aggregates both in the nucleus and in the cytoplasm of transfected cells, whereas the wild type protein localizes in the nucleus in a rather diffuse manner. Premature stop codons in FOXL2 have been considered so far as null alleles. However, we demonstrate here that such nonsense mutations may lead to the production of N terminally truncated proteins by re-initiation of translation downstream of the stop codon. Surprisingly, the truncated proteins strongly aggregate in the nucleus, partially localize in the cytoplasm and retain a fraction of the wild type protein. We also show that a complete deletion of the polyAla tract of FOXL2 induces a significant intranuclear aggregation. Our results enlarge the spectrum of mutations inducing FOXL2 aggregation. PMID- 16219627 TI - Development of MPS IVA mouse (Galnstm(hC79S.mC76S)slu) tolerant to human N acetylgalactosamine-6-sulfate sulfatase. AB - Mucopolysaccharidosis IVA (MPS IVA) is an autosomal recessive disease caused by N acetylgalactosamine-6-sulfate sulfatase (GALNS) deficiency. In recent studies of enzyme replacement therapy for animal models with lysosomal storage diseases, cellular and humoral immune responses to the injected enzymes have been recognized as major impediments to effective treatment. To study the long-term effectiveness and side effects of therapies in the absence of immune responses, we have developed an MPS IVA mouse model, which has many similarities to human MPS IVA and is tolerant to human GALNS protein. We used a construct containing both a transgene (cDNA) expressing inactive human GALNS in intron 1 and an active site mutation (C76S) in adjacent exon 2 and thereby introduced both the inactive cDNA and the C76S mutation into the murine Galns by targeted mutagenesis. Affected homozygous mice have no detectable GALNS enzyme activity and accumulate glycosaminoglycans in multiple tissues including visceral organs, brain, cornea, bone, ligament and bone marrow. At 3 months, lysosomal storage is marked within hepatocytes, reticuloendothelial Kupffer cells, and cells of the sinusoidal lining of the spleen, neurons and meningeal cells. The bone storage is also obvious, with lysosomal distention in osteoblasts and osteocytes lining the cortical bone, in chondrocytes and in the sinus lining cells in bone marrow. Ubiquitous expression of the inactive human GALNS was also confirmed by western blot using the anti-GALNS monoclonal antibodies newly produced, which resulted in tolerance to immune challenge with human enzyme. The newly generated MPS IVA mouse model should provide a good model to evaluate long-term administration of enzyme replacement. PMID- 16219628 TI - IVF results in de novo DNA methylation and histone methylation at an Igf2-H19 imprinting epigenetic switch. AB - Recent studies suggest that IVF and assisted reproduction technologies (ART) may result in abnormal genomic imprinting, leading to an increased frequency of Angelman syndrome (AS) and Beckwith-Weidemann syndrome (BWS) in IVF children. To learn how ART might alter the epigenome, we examined morulas and blastocysts derived from C57BL/6J X M. spretus F1 mice conceived in vivo and in vitro and determined the allelic expression of four imprinted genes: Igf2, H19, Cdkn1c and Slc221L. IVF-derived mouse embryos that were cultured in human tubal fluid (HTF) (Quinn's advantage) media displayed a high frequency of aberrant H19 imprinting, whereas in vivo and IVF embryos showed normal maternal expression of Cdkn1c and normal biallelic expression of Igf2 and Slc221L. Embryonic stem (ES) cells derived from IVF blastocysts also showed abnormal Igf2/H19 imprinting. Allele specific bisulphite PCR reveals abnormal DNA methylation at a CCCTC-binding factor (CTCF) site in the imprinting control region (ICR), as the normally unmethylated maternal allele acquired a paternal methylation pattern. Chromatin immunoprecipitation (ChIP) assays indicate an increase of lysine 4 methylation (dimethyl Lys4-H3) on the paternal chromatin and a gain in lysine 9 methylation (trimethyl Lys9-H3) on the maternal chromatin at the same CTCF-binding site. Our results indicate that de novo DNA methylation on the maternal allele and allele specific acquisition of histone methylation lead to aberrant Igf2/H19 imprinting in IVF-derived ES cells. We suggest that ART, which includes IVF and various culture media, might cause imprinting errors that involve both aberrant DNA methylation and histone methylation at an epigenetic switch of the Igf2-H19 gene region. PMID- 16219629 TI - Characterization of 17beta-hydroxysteroid dehydrogenase type 4 in human ovarian surface epithelial cells. AB - The human ovarian surface epithelium (hOSE) is a single layer of mesothelial-type primitive epithelial cells that are potential estrogen targets. It has been reported that hOSE cells can produce estrogen. However, the mechanisms that regulate estrogen level(s) in hOSE cells are not yet known. To elucidate the enzymes involved in these reactions, we examined gene expression of 17beta hydroxysteroid dehydrogenases (17beta-HSDs) in primary hOSE (POSE) and OSE2a cells using RT-PCR. We found that POSE cells and cells of the immortalized hOSE line, OSE2a, bidirectionally converted estrone (E1) and 17beta-estradiol (E2). Both cell types expressed mRNA for 17beta-HSD type 1 (17beta-HSD1), suggesting that the enzyme is involved in the E1 to E2 conversion. Interestingly, both cells expressed 17beta-HSD4 mRNA but not 17beta-HSD2 mRNA. We prepared an antibody against the carboxyl terminal of 17beta-HSD4 (anti-17beta-HSD4 antibody), which recognized the 80 and 48 kDa proteins in POSE and OSE2a cells based on immunoblot analysis. Furthermore, immunohistochemical study revealed the presence of 17beta HSD4 in hOSE cells in the human ovary. These results suggest that 17beta-HSD4 is involved in estrogen inactivation and may protect against an excessive accumulation of E2 in hOSE cells. PMID- 16219630 TI - Free school fruit--sustained effect 1 year later. AB - This study reports the effect of a school-randomized fruit and vegetable intervention consisting of a subscription to the Norwegian School Fruit Programme at no parental cost, and the Fruit and Vegetables Make the Marks (FVMM) educational programme, both delivered in the school year of 2001-02. Nine randomly chosen schools received the intervention and 10 schools served as control schools. Participating pupils completed questionnaires at baseline (September 2001), at Follow-up 1 (May-June 2002) and at Follow-up 2 (May 2003). A total of 517 pupils (84%; mean age, 11.3 years at baseline) participated in all three surveys. At both Follow-up 1 and Follow-up 2, strong intervention effects were observed for all-day fruit and vegetable intake (effect sizes were 0.6 and 0.5 portions, respectively). The sustained effect at Follow-up 2, 1 year after the end of the intervention, can partly be explained by greater participation rates in the School Fruit Programme (standard paid subscription). We conclude that the effects observed are most likely due to the no-cost subscription and not due to the FVMM educational programme, and that providing pupils with a piece of fruit or a vegetable at school at no cost for the parents is an effective strategy to increase school children's intake of fruit and vegetables. The effect is also sustained 1 year after the end of the no-cost subscription, providing increased health benefits. PMID- 16219631 TI - Outcome and process evaluation of a Norwegian school-randomized fruit and vegetable intervention: Fruits and Vegetables Make the Marks (FVMM). AB - This study reports the effect of the Fruits and Vegetables Make the Marks intervention, a school-based fruit and vegetable intervention consisting of a home economics classroom component and parental involvement and encouraged participation in the Norwegian School Fruit Programme, all delivered during the school year of 2001-02. Nine randomly chosen schools received the intervention and 10 schools served as control schools. Participating pupils completed questionnaires at baseline (September 2001), at Follow-up 1 (May-June 2002) and at Follow-up 2 (May 2003). A total of 369 pupils (69%; mean age, 11.3 years at baseline) participated in all three surveys. No effect of the intervention was found for intake of fruit and vegetables eaten at school or all day, neither at Follow-up 1 nor at Follow-up 2. On analysing the effects on potential mediators, significant differences between intervention and control groups were found for Awareness of the five-a-day recommendations only. The intervention programme was rated as very good by the teachers, and the pupils reported that they enjoyed it. However, the intervention failed to change fruit and vegetable intake, probably because it did not succeed in changing the pupils' preferences for or the accessibility of fruit and vegetables--the two strongest correlates of children's fruit and vegetable intake. PMID- 16219632 TI - Emerging role of KLF4 in human gastrointestinal cancer. AB - Recent analyses revealed that Kruppel-like factors (KLFs) play important roles in both normal development and carcinogenesis. Of the 16 known KLFs, KLF4 has been shown to be involved in the regulation of proliferation, differentiation and tumorigenesis of gastrointestinal tract epithelium. Clinical, experimental and mechanistic findings indicate that KLF4 is a bona fide tumor suppressor for both gastric and colorectal cancers. In this review, we summarize how this growing area of research has formed and the challenging new frontiers for better understanding of the oncogenic potential of the KLFs. PMID- 16219633 TI - An animal model for the rapid induction of tongue neoplasms in human c-Ha-ras proto-oncogene transgenic rats by 4-nitroquinoline 1-oxide: its potential use for preclinical chemoprevention studies. AB - Oral squamous cell carcinoma is one of the most common human neoplasms, and prevention of this malignancy requires a better understanding of its carcinogenesis process. To this end, we tried to establish an animal model using the human c-Ha-ras proto-oncogene-carrying transgenic (Tg) rats and the carcinogen 4-nitroquinoline 1-oxide (4-NQO). 4-NQO (20 p.p.m.) was administered to Tg and non-Tg rats for 8 weeks in their drinking water, and then the occurrence of tongue carcinogenesis was compared during the experimental period of 22 weeks. In addition, we determined the DNA ploidy in tongue lesions and examined the immunohistochemical expression of five biomarkers such as cyclin D1, glutathione S-transferase placental form, cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and beta-catenin. Next, the cancer chemopreventive effects of nimesulide, pioglitazone and a synthetic geranylated derivative, which have been reported to be inhibitors of tongue carcinogenesis, were examined in Tg rats treated with 4-NQO. Either during or after treatment with 4-NQO in the drinking water, tongue dysplasia and tumors were observed on the tongues of both Tg and non-Tg rats, with a greater incidence and multiplicity in Tg rats. Histopathologically, squamous cell dysplasia, papilloma and carcinoma with or without invasion were present in the tongue. Immunohistochemistry revealed that expression levels against five biomarkers increase with disease progression, and the changes correlated with those of the DNA ploidy pattern. Interestingly, a strong expression of COX-2, iNOS and beta-catenin was observed on the invasive front of squamous cell carcinomas. A subsequent chemoprevention study using Tg rats showed that the chemicals tested suppressed the occurrence of tongue carcinomas when they were administered after 4-NQO-exposure. These results may thus indicate that our 4-NQO-induced Tg rat tongue carcinogenesis model simulates many aspects of human oral carcinogenesis and it can be applied for an analysis of oral cancer development while also helping to identify potentially effective cancer chemopreventive agents against oral cancer. PMID- 16219634 TI - Meloxicam inhibits osteosarcoma growth, invasiveness and metastasis by COX-2 dependent and independent routes. AB - Cyclooxygenase-2 (COX-2) inhibitors exert antitumor activity via COX-2-dependent and independent pathways. We wished to evaluate the antitumor activity of meloxicam, a preferential COX-2 inhibitor, in osteosarcoma, the most common primary malignant bone tumor, and determine whether its antitumor effect is COX-2 dependent. COX-2 expression in the osteosarcoma cell lines MG-63, HOS and U2-OS was determined by real-time RT-PCR and western blotting. Subsequently, the inhibitory effects of meloxicam on osteosarcoma cell growth and invasiveness were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and matrigel invasion assays, respectively. Apoptotic activity was evaluated by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling staining and semi-quantification of Bax and Bcl-2 expression by real time RT-PCR and western blotting. Prostaglandin-E(2) (PGE(2)) production in the presence and absence of meloxicam was analyzed by enzyme immunoassay, and to determine whether the effects of meloxicam are COX-2-dependent or independent, PGE(2) was added to see if it reversed the effects of meloxicam. In addition, the effects of meloxicam on tumor growth and metastasis were evaluated in an in vivo mouse model using grafted LM-8 mouse osteosarcoma cells, together with immunohistochemical analysis for vascular endothelial growth factor in lung metastatic lesion. Meloxicam inhibited PGE(2) production, proliferation and invasiveness especially in MG-63 cells, which express relatively high levels of COX-2. Only high concentrations of meloxicam caused apoptosis and upregulated Bax mRNA and protein in MG-63 cell culture. In contrast, meloxicam did not induce apoptosis in HOS and U2-OS cells, expressing relatively low levels of COX-2. Exogenous PGE(2) reduced the effects of meloxicam on cell viability and invasiveness, but not its effect on Bax mRNA. In vivo, high doses of meloxicam suppressed LM-8 tumor growth and lung metastasis. Meloxicam, may have both COX-2-dependent and independent inhibitory actions on osteosarcoma. Its effects are more prominent in osteosarcoma cells that have relatively high levels of COX-2. PMID- 16219635 TI - Helicobacter pylori infection is associated with oxidatively damaged DNA in human leukocytes and decreased level of urinary 8-oxo-7,8-dihydroguanine. AB - Helicobacter pylori infection is responsible for inflammation, increased production of reactive oxygen species and oxidatively damaged DNA in the gastric mucosa. There is also evidence which suggests that H.pylori infection may lead to the development of several extragastroduodenal pathologies with reactive oxygen species involvement. In order to assess whether the infection may impose oxidatively damaged DNA not only in the target organ (stomach) but in other organs as well we decided, for the first time, to analyse the two kinds of oxidatively damaged DNA biomarkers: urinary excretion of 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodG) and 8-oxo-7,8-dihydroguanine (8-oxoGua) as well as the level of oxidatively damaged DNA in leukocytes. Using high performance liquid chromatography prepurification/gas chromatography with isotope dilution mass detection methodology, we examined the amount of oxidatively damaged DNA products excreted into urine and the amount of 8-oxodG in the DNA of leukocytes' (with the the HPLC/EC technique) in three groups of children: (i) control group, (ii) H.pylori infected children and (iii) children with gastritis where H.pylori infection was excluded. The levels of 8-oxodG in DNA isolated from leukocytes of H.pylori infected patients and in the group with gastritis without H.pylori infection were significantly higher than in DNA isolated from the control group. The mean level of 8-oxoGua in urine samples of children infected with H.pylori was significantly lower than in the urine of the group with gastritis without H.pylori infection. The data suggest that inflammation itself, not just H.pylori infection, is responsible for the observed rise of 8-oxodG level in leukocytes. However, the observed decrease in the level of modified base in urine seems to be specific for H.pylori infection and possibly linked with nitric oxide mediated inhibition of a key base excision repair enzyme (human 8-oxo-7, 8-dihydroguanine glycosylase) responsible for the repair of 8-oxo-7,8-dihydroguanine. PMID- 16219636 TI - Oligonucleotide microarray analysis of distinct gene expression patterns in colorectal cancer tissues harboring BRAF and K-ras mutations. AB - Various types of human cancers harbor BRAF somatic mutations, leading researchers to seek molecular targets for BRAF inhibitors. A mutually exclusive relationship has been observed between the BRAF-V600E mutation and K-ras mutations, suggesting that the BRAF-V600E mutation may differ from the other BRAF mutant types. Here, we used microarray analysis to examine differences between the BRAF and K-ras mutant colorectal samples and within the BRAF group (V600E versus non-V600E), in the hope that the identified gene sets could form the basis for new target development. Eleven colorectal cancers (CRCs) with BRAF mutations and nine with K ras mutations were examined by high-density microarray analysis. We also tested whether other significant genetic or clinical status involved in CRC development, such as APC and TP53 mutations, MSI and TNM-Duke's staging, were related with the observed BRAF- or K-ras associated expression profiles. Unsupervised two-way hierarchical clustering and multidimensional scaling revealed that the differentially expressed genes clustered according to the mutation status of BRAF and K-ras, and that samples with the BRAF-V600E and non-V600E mutants could be distinguished from each other by gene profiling. Examination of TNM-Duke's staging, MSI and mutations in APC and TP53 revealed that these significant mutations could not account for the hierarchical clustering results observed in our study. We herein identified distinct gene expression patterns and gene sets that may form the basis for identification of BRAF-targeting molecules or provide researchers with a better understanding of the molecular pathogenesis underlying RAS-RAF signaling. PMID- 16219638 TI - The positional effect of Ebeta on Vbeta genes of TCRbeta chain in the ordered rearrangement and allelic exclusion. AB - In the TCRbeta gene locus, the Vbeta, Dbeta and Jbeta gene segments are assembled in a tightly ordered manner. To investigate the positional effects of TCRbeta enhancer (Ebeta) on the recombination processes of the Vbeta genes, we utilized beta(LD) mice lacking 70% of the TCRbeta locus, leaving four Vbeta genes at the 5' side and, consequently, the Vbeta10 gene moves into the Ebeta regulatory region. In this mutant mouse, the Vbeta10 gene showed direct Vbeta-to-Dbeta and Vbeta-to-Jbeta recombination, although the Dbeta-to-Jbeta joining was still predominant. Interestingly, these two aberrant recombination processes were barely suppressed when beta(LD) mice were crossed with TCRbeta transgenic mice, whereas V(D)J recombination of the Vbeta10 gene was sufficiently suppressed. These results suggest that the positional effects of Ebeta on the Vbeta genes may enable the recombination potential to increase prior to Dbeta-to-Jbeta joining and that such aberrant recombination may be free from allelic suppression. PMID- 16219637 TI - Regulation of stromal cell cyclooxygenase-2 in the ApcMin/+ mouse model of intestinal tumorigenesis. AB - Cyclooxygenase-2 (Cox-2) is expressed predominantly by stromal cells in intestinal adenomas from the Apc(Min/+) mouse model of familial adenomatous polyposis. We investigated the mechanistic basis of stromal cell Cox-2 expression in Apc(Min/+) mouse adenomas, as well as Cox-2 expression and activity in histologically normal (HN) Apc(Min/+) mouse intestine, in order to gain further insights into regulation of Cox-2 as a potential chemoprevention target. Upregulation of Cox-2 in intestinal tumours is not an intrinsic feature of Apc(Min/+) macrophages as bone marrow-derived Apc(Min/+) macrophages did not exhibit an abnormality in Cox-2 expression or activity. Intestinal permeability to lactulose or mannitol was similar in Apc(Min/+) mice and wild-type littermates, implying that macrophage activation by luminal antigen is unlikely to explain stromal cell Cox-2 induction. Moreover, stromal cells exhibited differential expression of Cox-2 and inducible nitric oxide synthase, suggesting 'alternative' (M2) rather than 'classical' (M1) macrophage activation. Flow cytometric sorting of isolated stromal mononuclear cells (SMNCs), on the basis of M-lysozyme and specific macrophage marker expression, demonstrated that macrophages, neutrophils and non-myelomonocytic cells all contributed to lamina propria prostaglandin (PG) E(2) synthesis. However, the majority of PGE(2) synthesis by macrophages was via a Cox-2-dependent pathway compared with predominant Cox-1-derived PGE(2) production by non-myelomonocytic cells. SMNCs from HN Apc(Min/+) intestinal mucosa exhibited similar levels of Cox-2 mRNA and protein, but produced more Cox-2-derived PGE(2) than wild-type cells at 70 days of age. There was an age-dependent decline in PGE(2) synthesis by Apc(Min/+) SMNCs, despite tumour progression. These data suggest that other Cox-2 independent factors also control PGE(2) levels during Apc(Min/+) mouse intestinal tumorigenesis. Regulation of macrophage Cox-2 expression and other steps in PGE(2) synthesis (e.g. PGE synthase) are valid targets for novel chemoprevention strategies that could minimize or avoid systemic COX-2 inhibition. PMID- 16219639 TI - Physical and functional interactions between STAT3 and ZIP kinase. AB - Signal transducer and activator of transcription 3 (STAT3) is a latent cytoplasmic transcription factor that can be activated by cytokines and growth factors. It plays important roles in cell growth, apoptosis and cell transformation, and is constitutively active in a variety of tumor cells. In this study, we provide evidence that zipper-interacting protein kinase (ZIPK) interacts physically with STAT3. ZIPK specifically interacted with STAT3, and did not bind to STAT1, STAT4, STAT5a, STAT5b or STAT6. ZIPK phosphorylated STAT3 on serine 727 (Ser727) and enhanced STAT3 transcriptional activity. Small interfering RNA-mediated reduction of ZIPK expression decreased leukemia inhibitory factor (LIF)- and IL-6-induced STAT3-dependent transcription. Furthermore, LIF- and IL-6-mediated STAT3 activation stimulated ZIPK activity. Taken together, our data suggest that ZIPK interacts with STAT3 within the nucleus to regulate the transcriptional activity of STAT3 via phosphorylation of Ser727. PMID- 16219640 TI - Increased expression of aggrecan and biglycan mRNA in Achilles tendinopathy. AB - OBJECTIVES: To determine the expression of mRNA encoding the proteoglycans aggrecan, versican, biglycan and decorin in mid-tendon samples of chronic painful Achilles tendinopathy and ruptured Achilles tendons, compared with normal tendons. METHODS: Total RNA isolated from frozen tendon samples (14 normal, 13 painful, 14 ruptured) was assayed by relative quantitative reverse transcription polymerase chain reaction for aggrecan, versican, biglycan and decorin mRNA, normalized using 18S rRNA. Differences between sample groups were tested by univariate analysis of variance with age as co-variate. RESULTS: In normal tendon samples expression of each of the proteoglycan mRNA decreased with increasing age. Decorin mRNA was the most highly-expressed of the proteoglycan mRNA, while versican mRNA expression was higher (3.8-fold) than that of aggrecan. In painful tendinopathy both aggrecan and biglycan mRNA expression increased (more than 10 fold and 5-fold, respectively) compared with normal tendon samples, but levels of versican and decorin mRNA were not significantly changed. In ruptured tendons the levels of aggrecan, biglycan and versican mRNA were not changed compared with normal tendon samples, but decorin mRNA decreased markedly. CONCLUSIONS: Increased aggrecan and biglycan mRNA expression in painful tendinopathy resembles the pattern in fibrocartilaginous regions of tendon, and may reflect an altered mechanical environment at the site of the lesion. Increased aggrecan mRNA expression may underlie the increase in glycosaminoglycan observed in painful tendinopathy. PMID- 16219641 TI - Levels of F2-isoprostanes in systemic sclerosis: correlation with clinical features. AB - OBJECTIVE: Oxidative stress may be one of the important complex pathogenetic mechanisms that lead to damage in scleroderma; free radicals may provoke endothelial injury, fibroblast proliferation and fragmentation of autoantigens favouring induction of autoantibodies. The present study investigates the oxidant status in scleroderma patients by measuring the urinary concentration of 8 isoprostaglandin-F2alpha, an F2-isoprostane, and a product of free radical mediated peroxidation of arachidonic acid. METHODS: Forty-three scleroderma patients (42 women and 1 man, mean age 54.1 yr, mean disease duration 9.0 yr) underwent clinical evaluation and instrumental investigations in order to assess skin, vascular, lung and heart involvement. Von Willebrand factor was evaluated as marker of vascular dysfunction in 36 out of the 43 cases. The urinary level of 8-isoprostaglandin-F2alpha was measured in all scleroderma patients and in the 43 age- and sex-matched healthy controls. RESULTS: Urinary levels of 8 isoprostaglandin-F2alpha were higher in scleroderma patients than in the healthy control group (341.7 vs 147.6 pg/mg creatinine; P < 0.001). Values of 8 isoprostaglandin-F2alpha were strongly correlated with the nailfold videocapillaroscopy pattern and lung involvement (P = 0.002 and 0.003, respectively), showing increasing levels with the progression of pulmonary severity. Correlation between 8-isoprostaglandin-F2alpha level and von Willebrand factor narrowly failed to reach statistical significance (P = 0.05). There was no correlation between 8-isoprostaglandin-F2alpha concentration and disease activity, vascular, skin and heart involvement, disease pattern or autoantibody profile. CONCLUSIONS: Our study further supports the role of oxidant stress in the pathogenesis of scleroderma, showing a strong correlation between a marker of free radical damage with both the severity of lung involvement and the videocapillaroscopic patterns. PMID- 16219642 TI - Pneumococcal sepsis after autosplenectomy in a girl with systemic lupus erythematosus. PMID- 16219643 TI - A case of eosinophilic enteritis and rheumatoid arthritis. PMID- 16219644 TI - Significance of SAA1.3 allele genotype in Japanese patients with amyloidosis secondary to rheumatoid arthritis. AB - OBJECTIVE: To clarify the clinical significance of the SAA1.3 allele in the development and outcome of AA amyloidosis in Japanese patients with rheumatoid arthritis (RA). METHODS: One hundred and twenty RA patients (60 alive and 60 dead) fulfilling the 1987 ACR criteria and 62 RA patients with biopsy-confirmed amyloid A (AA) amyloidosis (36 alive and 26 dead) were enrolled. The SAA1 genotypes were determined by PCR-based restriction fragment length polymorphism. To predict the clinical outcome of AA amyloidosis, we investigated characteristics and survival, focusing on the SAA1.3 allele retrospectively. RESULTS: The SAA1.3 allele genotype was not only a risk factor for the association of AA amyloidosis but also a poor prognostic factor for the development of AA amyloidosis (P=0.015). Both the association of AA amyloidosis arising early in the RA disease course and symptomatic variety and severity were found in amyloidotic patients with the SAA1.3 allele. The presenting factors adversely influenced were age (P=0.001), lowered serum albumin (P=0.001) and creatinine concentration (P=2.14 x 10(-5)). Renal involvement was associated with poor survival in patients with AA amyloidosis (P=0.011) and the presence of cardiac involvement was likely to be a risk factor for survival (P=0.062). The rate of the causes of death in respect to the category of infection, gastrointestinal diseases, and renal failure was higher in patients with AA amyloidosis than in those without amyloidosis, gastrointestinal diseases and renal failure. Cyclophosphamide was found to be superior to methotrexate in the management of RA patients with AA amyloidosis. CONCLUSION: Our data support the fact that homozygosity for the SAA1.3 allele is a univariate predictor of survival in addition to a risk factor for the association of AA amyloidosis adversely influencing the outcome in Japanese RA patients. Renal involvement is a pivotal clinical manifestation in the development of AA amyloidosis, as is likely to be cardiac involvement in AA amyloidosis secondary to RA. PMID- 16219647 TI - BiDil: race medicine or race marketing? AB - Recent Food and Drug Administration (FDA) approval of the first drug with a race specific indication has fueled the controversy over the meaning of race and ethnicity and raised questions over whether this move should be seen as an advance or a setback in the struggle to address disparities in health status associated with race. The drug, BiDil, combines two generics long recognized as benefiting patients with heart failure, irrespective of race or ethnicity. The push to bring these drugs to market as a race-specific treatment was motivated by the culiarities of U.S. patent law and willingness exploit race to gain commercial and regulatory advantage. PMID- 16219645 TI - Low bone density and low serum levels of soluble RANK ligand are associated with severe arterial calcification in patients with Takayasu arteritis. AB - OBJECTIVE: Takayasu arteritis (TA) is a chronic inflammatory disorder affecting the aorta and its branches. Vascular calcification has been described in 29-54% of cases of TA, although its aetiology remains unknown. Recently the osteoprotegerin/RANKL/RANK system has emerged as an important contributing factor to atherogenesis and osteogenesis. Our aim is to investigate the association between vascular calcification, bone mineral density (BMD) and the osteoprotegerin/RANK/RANKL system in TA. METHODS: Thirty pre-menopausal female TA patients and 30 age- and sex-matched controls were studied. BMD was measured by dual X-ray absorptiometry. Arterial calcification in TA patients was analysed by computed tomography in thoracic and abdominal sites. Serum levels of osteoprotegerin and soluble receptor activator of nuclear factor kappaB ligand (sRANKL) were quantified by enzyme-linked immunosorbent assay. RESULTS: Patients with severe arterial calcification showed lower BMD values than controls in lumbar spine (0.965 +/- 0.055 vs 1.126 +/- 0.153 g/cm2, P = 0.009) and total body (0.993 +/- 0.065 vs 1.085 +/- 0.082 g/cm2, P = 0.019). In contrast, TA patients without calcification presented BMD values similar to controls (P > 0.05). Interestingly, lower serum levels of sRANKL (1.89 +/- 2.35 vs 2.80 +/- 2.23 pg/ml, P = 0.031) and a longer disease duration (12.20 +/- 6.61 vs 3.56 +/- 5.33 yr, P = 0.004) were observed in TA patients with severe calcification compared with patients without calcification. CONCLUSIONS: Severe arterial calcification in TA is associated with low values of BMD and sRANKL, reinforcing the possible link between bone and vascular disease. PMID- 16219648 TI - The case of BiDil: a policy commentary on race and genetics. AB - The Food and Drug Administration (FDA) approval of BiDil unleashed a vigorous commentary, mostly critical of the decision. The FDA was soon caught between biologists, who see research and clinical utility in using racial classifications, and social scientists (and many politicians), who fear the adversities of greater discrimination. Analyses rarely addressed the practical factors the FDA had to consider in reaching a decision. Much of the literature simply assigned the question to the domain of racial politics, failing to consider the ethics of professional care, the Hippocratic oath, and the marketplace efficiency in moving drugs quickly to those who might most benefit. PMID- 16219649 TI - If at first you don't succeed try ... a new target in the treatment of angina. PMID- 16219650 TI - Diet and inflammation: a link to metabolic and cardiovascular diseases. AB - Both epidemiological studies and intervention trials support an important role of diet in reducing the risk of a variety of chronic diseases, including cardiovascular disease, and overall mortality. We discuss available evidence indicating that the generation of a pro-inflammatory milieu might be one mechanism through which unhealthy diets are linked to metabolic and cardiovascular diseases. In practical terms, fully understanding the link between diet and inflammation holds the premise to elucidate the mechanisms by which dietary patterns improve cardiovascular health. PMID- 16219651 TI - Mirror, mirror on the wall: the quest for the earliest marker of myocardial ischaemia. PMID- 16219652 TI - Managing patients with heart failure: the reality of clinical practice. PMID- 16219653 TI - Heart failure in elderly patients with chronic obstructive pulmonary disease. PMID- 16219654 TI - First comes diagnosis then comes treatment: an under-appreciated paradigm in atrial fibrillation management. PMID- 16219655 TI - Left atrial ablation pendulum swinging back towards the pulmonary veins. PMID- 16219656 TI - Is the jury out? Class specific differences on coronary outcomes with ACE inhibitors and ARBs: insight from meta-analysis and The Blood Pressure Lowering Treatment Trialists' Collaboration. PMID- 16219657 TI - Epidemiology of non-ST elevation acute coronary syndromes in the Italian cardiology network: the BLITZ-2 study. AB - AIMS: Acute coronary syndromes without ST-segment elevation (NSTEACS) represent an increasingly frequent cause of hospital admission. The BLITZ-2 study was planned to survey the epidemiology and management strategies of NSTEACS in the Italian cardiological network. METHODS AND RESULTS: The study included 1888 patients with NSTEACS in 275 hospitals in 3 weeks. At admission, almost 20% of patients showed clinical signs of heart failure, half showed ST-segment depression, and half showed any positive biochemical myocardial necrosis marker. Patients admitted to hospitals without CathLab (n=973) were older (P=0.0005) and with higher Killip class on admission (P<0.0001) when compared with those admitted to hospitals with CathLab (n=915). During index hospitalization, 76% of the patients initially admitted to hospitals with invasive capability underwent coronary angiography and 39% percutaneous coronary intervention when compared with 39 and 17.2% of those admitted to hospitals without CathLab (P<0.001). Overall, 30-day mortality was 2.4% (2.0% in patients with invasive capability vs. 2.9% in hospitals without CathLab, P=0.2). Cardiac ischaemic events at 30 days (recurrent MI, recurrent angina, and re-hospitalization for ACS) were significantly higher in the group of patients admitted to hospitals without CathLab (OR 1.71, 95% CI 1.24-2.35). However, after multivariable adjustment, only advanced age (OR 1.043, 95% CI 1.021-1.065, P<0.0001) and Killip class >1 (OR 1.633, 95% CI 1.020-2.614, P=0.04) resulted in independent predictors of death, in-hospital MI, and re-admission for ACS, whereas the absence of an on site CathLab did not predict an adverse outcome (OR 1.104, 95% CI 0.734-1.660). CONCLUSION: According to this, the nationwide registry outcome is only marginally influenced by invasive procedures. Contemporary management of patients with NSTEACS in Italy is primarily driven by resource availability. PMID- 16219658 TI - Predictors of mortality and morbidity in patients with chronic heart failure. AB - AIMS: We aimed to develop prognostic models for patients with chronic heart failure (CHF). METHODS AND RESULTS: We evaluated data from 7599 patients in the CHARM programme with CHF with and without left ventricular systolic dysfunction. Multi-variable Cox regression models were developed using baseline candidate variables to predict all-cause mortality (n=1831 deaths) and the composite of cardiovascular (CV) death and heart failure (HF) hospitalization (n=2460 patients with events). Final models included 21 predictor variables for CV death/HF hospitalization and for death. The three most powerful predictors were older age (beginning >60 years), diabetes, and lower left ventricular ejection fraction (EF) (beginning <45%). Other independent predictors that increased risk included higher NYHA class, cardiomegaly, prior HF hospitalization, male sex, lower body mass index, and lower diastolic blood pressure. The model accurately stratified actual 2-year mortality from 2.5 to 44% for the lowest to highest deciles of predicted risk. CONCLUSION: In a large contemporary CHF population, including patients with preserved and decreased left ventricular systolic function, routine clinical variables can discriminate risk regardless of EF. Diabetes was found to be a surprisingly strong independent predictor. These models can stratify risk and help define how patient characteristics relate to clinical course. PMID- 16219659 TI - Direct thrombin inhibitors in acute coronary syndrome patients undergoing percutaneous coronary intervention: special effects in selected patients? PMID- 16219660 TI - CFTR, chloride concentration and cell volume: could mammalian protein histidine phosphorylation play a latent role? AB - A considerable body of evidence indicates that the intracellular chloride concentration ([Cl-]i) is an important regulatory signal in epithelial ion transport. [Cl-]i regulates the open channel probability of sodium and chloride channels, the rate of chloride channel recycling to the apical membrane, cell volume homeostasis, the activity of sodium-coupled chloride entry pathways and G protein activity. Cell volume goes awry in epithelial cells bearing mutant forms of the cystic fibrosis (CF) transmembrane conductance regulator protein (CFTR); however, the pathways that mediate this [Cl-]i effect at the apical membrane of polarized epithelia are unknown. Recently, we proposed a mechanism for the transduction of in vitro chloride concentration into a phosphorylation signal to proteins within the apical membrane of respiratory epithelia. Our studies show that an apically enriched plasma membrane fraction from a variety of species, including sheep, human and mouse airway, contains at least two membrane-bound protein kinases which exhibit a number of novel properties. Firstly, the phosphate is located on histidine residues within different families of proteins; one kinase(s) utilizes GTP rather than ATP as a phosphate donor and each kinase has its own unique profile of membrane protein phosphorylation (which itself varies with anion species). Secondly, both kinases mediate Cl- -dependent phosphorylation of an apical membrane protein around the established physiological values for [Cl-]i in airway epithelial cells ( approximately 40 mM); associated phosphatases also alter the net phosphoprotein profile of the apical membrane. These findings are reviewed and their potential roles explored in relation to the pathogenesis of CF using the control of cell volume as a model for disrupted cellular function in CF-affected epithelia. PMID- 16219661 TI - The Drosophila tweety family: molecular candidates for large-conductance Ca2+ activated Cl- channels. AB - Calcium-activated chloride currents (I(Cl(Ca))) can be recorded in almost all cells, but the molecular identity of the channels underlying this Cl- conductance is still incompletely understood. Here, I report that tweety, a gene located in Drosophila flightless, possesses five or six transmembrane segments, and that a human homologue of tweety (hTTYH3) is a novel large-conductance Ca2+-activated Cl channel, while the related gene, hTTYH1, is a swelling-activated Cl- current. hTTYH3 is expressed in excitable tissues, including the heart, brain and skeletal muscle, whereas hTTYH1 is expressed mainly in the brain. Expression of hTTYH3 in CHO cells generated a unique Cl- current activated by an increase in the intracellular Ca2+ concentration. The hTTYH3-induced Cl- current had a linear current-voltage (I-V) relationship, a large single-channel conductance (260 pS) and the anion permeability sequence I- > Br- > Cl-. Like native Ca2+-activated Cl channels, the hTTYH3 channel showed complex gating kinetics and voltage dependent inactivation, and was dependent on micromolar intracellular Ca2+ concentration. Expression in CHO cells of an hTTYH1 splice variant that lacks the C-terminal glutamate-rich domain of hTTYH1 (hTTYH1sv) generated a swelling activated Cl- current. I conclude that investigation of the tweety family will provide important information about large-conductance Cl- channel molecules. PMID- 16219662 TI - Glycosylphosphatidylinositol-specific phospholipase D influences triglyceride rich lipoprotein metabolism. AB - Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is a minor HDL associated protein. Because many minor HDL-associated proteins exchange between different lipoprotein classes during the postprandial state and are also involved in triglyceride (TG) metabolism, we hypothesized that GPI-PLD may play a role in the metabolism of TG-rich lipoproteins. To test this hypothesis, we examined the distribution of GPI-PLD among lipoprotein classes during a fat tolerance test in C57BL/6 and LDL receptor-deficient (LDLR(-/-)) mice fed either a chow or high fructose diet. In the fasting state in wild-type mice fed a chow diet, GPI-PLD was only present in HDL, whereas in LDLR(-/-) mice GPI-PLD was present in HDL and intermediate-density lipoproteins (IDL)/LDL. During the fat tolerance test, there was no change in total serum GPI-PLD levels in either model; however, a significant amount of GPI-PLD appeared in both VLDL (0.5-1% of total GPI-PLD) and IDL/LDL (5-10% of total GPI-PLD) in both models. The high-fructose diet increased both fasting and postprandial TG and serum GPI-PLD levels in both strains as well as the amount of GPI-PLD in VLDL. To determine whether GPI-PLD plays a direct role in TG metabolism, we increased liver GPI-PLD expression in C57BL/6 mice by adenovirus-mediated gene transfer, which resulted in a sevenfold increase in serum GPI-PLD levels. This change was associated with an increase in fasting (30%) and postprandial TG (50%) and a twofold reduction in TG-rich lipoprotein catabolism compared with saline or control adenovirus-treated mice. These studies demonstrate that GPI-PLD affects serum TG levels by altering catabolism of TG rich lipoproteins. PMID- 16219663 TI - Acute hypoxia decreases plasma VEGF concentration in healthy humans. AB - Vascular endothelial growth factor (VEGF) is known to be upregulated by hypoxia in vitro. However, in vivo data about VEGF regulation in chronic hypoxic diseases are conflicting. We investigated the effects of hypoxia on plasma VEGF concentration in healthy subjects. To control known confounders, such as insulin, glucose concentrations, or exercise, hypoxic effects on VEGF were studied during experimentally clamping glucose concentrations at rest. In a double-blind crossover study design, we induced hypoxia for 30 min by decreasing oxygen saturation to 75% (vs. normoxic control) in 14 healthy men. Plasma VEGF concentration was determined at baseline, immediately after hypoxia had ended, and after a further 150 min. Levels of its soluble (s)Flt-1 receptor were assessed at baseline and at the end of the clamp. In parallel, catecholamine and cortisol levels were monitored. To investigate potential effects of glucose administration on the release of VEGF, we performed a third session, reducing glucose infusion for 30 min while serum insulin was held stable thereby inducing hypoglycemia. Hypoxia decreased VEGF levels compared with the normoxic control (P<0.05). VEGF concentrations increased during hypoglycemia (P<0.02) but were comparable to the normoglycemic control at the end of the clamp (P>0.80). sFlt-1 receptor concentration remained unchanged during hypoxia and hypoglycemia compared with control (both P>0.4). Epinephrine concentration (P<0.01) increased upon hypoxia, whereas norepinephrine and cortisol did not change. Contrary to in vitro studies, in healthy humans hypoxia decreases plasma VEGF concentration, suggesting that systemic VEGF concentration may be differently regulated than the expression on cellular basis. PMID- 16219664 TI - Myofibrillar protein synthesis in myostatin-deficient mice. AB - Either increased protein synthesis or prolonged protein half-life is necessary to support the excessive muscle growth and maintenance of enlarged muscles in myostatin-deficient mice. This issue was addressed by determining in vivo rates of myofibrillar protein synthesis in mice with constitutive myostatin deficiency (Mstn(DeltaE3/DeltaE3)) or normal myostatin expression (Mstn(+/+)) by measuring tracer incorporation after a systemic flooding dose of l-[ring (2)H(5)]phenylalanine. At 5-6 wk of age, Mstn(DeltaE3/DeltaE3) mice had increased muscle mass (40%), fractional rates of myofibrillar synthesis (14%), and protein synthesis per whole muscle (60%) relative to Mstn(+/+) mice. With maturation, fractional rates of synthesis declined >50% in parallel with decreased DNA and RNA [total, 28S rRNA, and poly(A) RNA] concentrations in muscle. At 6 mo of age, Mstn(DeltaE3/DeltaE3) mice had even greater increases in muscle mass (90%) and myofibrillar synthesis per muscle (85%) relative to Mstn(+/+) mice, but the fractional rate of synthesis was normal. Estimated myofibrillar protein half-life was not affected by myostatin deficiency. Muscle DNA concentrations were reduced in both young and mature Mstn(DeltaE3/DeltaE3) mice, whereas RNA concentrations were normal, so the ratio of RNA to DNA was approximately 30% greater than normal in Mstn(DeltaE3/DeltaE3) mice. Thus the increased protein synthesis and RNA content per muscle in myostatin-deficient mice cannot be explained entirely by an increased number of myonuclei. PMID- 16219665 TI - Energy state of the liver during short-term and exhaustive exercise in C57BL/6J mice. AB - A portal venous 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside infusion that results in hepatic 5-aminoimidazole-4-carboxamide-1-beta-D-ribosyl-5 monophosphate (ZMP) concentrations of approximately 4 micromol/g liver increases hepatic glycogenolysis and glucose output. ZMP is an AMP analog that mimics the regulatory actions of this nucleotide. The aim of this study was to measure hepatic AMP concentrations in response to increasing energy requirements to test the hypothesis that AMP achieves concentrations during exercise, consistent with a role in stimulation of hepatic glucose metabolism. Male C57BL/6J mice (27.4+/- 0.4 g) were subjected to 35 min of rest [sedentary (SED), n=8], underwent short term (ST, 35 min) moderate (20 m/min, 5% grade) exercise (n=8), or underwent treadmill exercise under similar conditions but until exhaustion (EXH, n=8). Hepatic AMP concentrations were 0.82+/- 0.05, 1.17+/- 0.11, and 2.52+/- 0.16 micromol/g liver in SED, ST, and EXH mice, respectively (P< 0.05). Hepatic energy charge was 0.66+/- 0.01, 0.58+/- 0.02, and 0.33+/- 0.22 in SED, ST, and EXH mice, respectively (P< 0.05). Hepatic glycogen was 11.6+/- 1.0, 8.8+/- 2.2, and 0.0+/- 0.1 mg/g liver in SED, ST, and EXH mice, respectively (P< 0.05). Hepatic AMPK (Thr(172)) phosphorylation was 1.00+/- 0.14, 1.96+/- 0.16, and 7.44+/- 0.63 arbitrary units in SED, ST, and EXH mice, respectively (P< 0.05). Thus exercise increases hepatic AMP concentrations. These data suggest that the liver is highly sensitive to metabolic demands, as evidenced by dramatic changes in cellular energy indicators (AMP) and sensors thereof (AMP-activated protein kinase). In conclusion, AMP is sensitively regulated, consistent with it having an important role in hepatic metabolism. PMID- 16219666 TI - Human duodenal enteroendocrine cells: source of both incretin peptides, GLP-1 and GIP. AB - Among the products of enteroendocrine cells are the incretins glucagon-like peptide-1 (GLP-1, secreted by L cells) and glucose-dependent insulinotropic peptide (GIP, secreted by K cells). These are key modulators of insulin secretion, glucose homeostasis, and gastric emptying. Because of the rapid early rise of GLP-1 in plasma after oral glucose, we wished to definitively establish the absence or presence of L cells, as well as the relative distribution of the incretin cell types in human duodenum. We confirmed the presence of proglucagon and pro-GIP genes, their products, and glucosensory molecules by tissue immunohistochemistry and RT-PCR of laser-captured, single duodenal cells. We also assayed plasma glucose, incretin, and insulin levels in subjects with normal glucose tolerance and type 2 diabetes for 120 min after they ingested 75 g of glucose. Subjects with normal glucose tolerance (n=14) had as many L cells (15+/ 1), expressed per 1,000 gut epithelial cells, as K cells (13+/-1), with some containing both hormones (L/K cells, 5+/-1). In type 2 diabetes, the number of L and L/K cells was increased (26+/-2; P<0.001 and 9+/-1; P < 0.001, respectively). Both L and K cells contained glucokinase and glucose transporter-1, -2, and -3. Newly diagnosed type 2 diabetic subjects had increased plasma GLP-1 levels between 20 and 80 min, concurrently with rising plasma insulin levels. Significant coexpression of the main incretin peptides occurs in human duodenum. L and K cells are present in equal numbers. New onset type 2 diabetes is associated with a shift to the L phenotype. PMID- 16219667 TI - Identification of fatty acid translocase on human skeletal muscle mitochondrial membranes: essential role in fatty acid oxidation. AB - Fatty acid translocase (FAT/CD36) is a transport protein with a high affinity for long-chain fatty acids (LCFA). It was recently identified on rat skeletal muscle mitochondrial membranes and found to be required for palmitate uptake and oxidation. Our aim was to identify the presence and elucidate the role of FAT/CD36 on human skeletal muscle mitochondrial membranes. We demonstrate that FAT/CD36 is present in highly purified human skeletal mitochondria. Blocking of human muscle mitochondrial FAT/CD36 with the specific inhibitor sulfo-N succimidyl-oleate (SSO) decreased palmitate oxidation in a dose-dependent manner. At maximal SSO concentrations (200 muM) palmitate oxidation was decreased by 95% (P<0.01), suggesting an important role for FAT/CD36 in LCFA transport across the mitochondrial membranes. SSO treatment of mitochondria did not affect mitochondrial octanoate oxidation and had no effect on maximal and submaximal carnitine palmitoyltransferase I (CPT I) activity. However, SSO treatment did inhibit palmitoylcarnitine oxidation by 92% (P<0.001), suggesting that FAT/CD36 may be playing a role downstream of CPT I activity, possibly in the transfer of palmitoylcarnitine from CPT I to carnitine-acylcarnitine translocase. These data provide new insight regarding human skeletal muscle mitochondrial fatty acid (FA) transport, and suggest that FAT/CD36 could be involved in the cellular and mitochondrial adaptations resulting in improved and/or impaired states of FA oxidation. PMID- 16219668 TI - Lipid metabolism response to a single, prolonged bout of endurance exercise in healthy young men. AB - To discover the alterations in lipid metabolism linked to postexercise hypotriglyceridemia, we measured lipid kinetics, lipoprotein subclass distribution and lipid transfer enzymes in seven healthy, lean, young men the day after 2 h of cycling and rest. Compared with rest, exercise increased fatty acid rate of appearance and whole body fatty acid oxidation by approximately 65 and 40%, respectively (P < 0.05); exercise had no effect on VLDL-triglyceride (TG) secretion rate, increased VLDL-TG plasma clearance rate by 40 +/- 8%, and reduced VLDL-TG mean residence time by approximately 40 min and VLDL-apolipoprotein B-100 (apoB-100) secretion rate by 24 +/- 8% (all P < 0.05). Exercise also reduced the number of VLDL but almost doubled the number of IDL particles in plasma (P < 0.05). Muscle lipoprotein lipase content was not different after exercise and rest, but plasma lipoprotein lipase concentration increased by approximately 20% after exercise (P < 0.05). Plasma hepatic lipase and lecithin:cholesterol acyltransferase concentrations were not affected by exercise, whereas cholesterol ester transfer protein concentration was approximately 10% lower after exercise than after rest (P = 0.052). We conclude that 1) greater fatty acid availability after exercise does not stimulate VLDL-TG secretion, probably because of the increase in fatty acid oxidation and possibly also fatty acid use for restoration of tissue TG stores; 2) reduced secretion of VLDL-apoB-100 lowers plasma VLDL particle concentration; and 3) increased VLDL-TG plasma clearance maintains low plasma TG concentration but is not accompanied by similar increases in subsequent steps of the delipidation cascade. Acutely, therefore, the cardioprotective lowering of plasma TG and VLDL concentrations by exercise is counteracted by a proatherogenic increase in IDL concentration. PMID- 16219669 TI - Glucocorticoids differentially regulate expression of duodenal and renal calbindin-D9k through glucocorticoid receptor-mediated pathway in mouse model. AB - Dexamethasone (Dex) is a member of the glucocorticoids (GCs), and is broadly used as an anti-inflammatory medication. Continuous administration with GCs induces adverse effects and suffering in humans (i.e., osteoporosis) due to negative calcium balance derived from low re- and absorption in the duodenum and kidney. A cytosolic calcium-binding protein, calbindin-D9k (CaBP-9k), is dominantly expressed in the renal and intestinal tissues involved in calcium re- and absorption and plays an active role in calcium transport. In the present study, we employed adrenalectomized (ADX) and sham-treated (Sham) male mice to examine the effect of Dex on CaBP-9k gene expression in the duodenum and kidney. Dex significantly reduced the levels of duodenal CaBP-9k mRNA and protein, and it restored ADX-induced decrease in renal CaBP-9k protein compared with the level of Sham control. Dex treatment increased calcium and phosphate levels in the sera of both Sham and ADX mice. In a time course experiment, Dex significantly decreased duodenal CaBP-9k at the transcriptional and translational levels at 3 days, whereas it temporarily increased CaBP-9k mRNA and protein levels at 12 and 24 h. Altered CaBP-9k expression by Dex was completely reversed by mifepristone, an antagonist for the GC receptor (GR). In addition, duodenal CaBP-9k and GR were colocalized on the enterocyte (duodenocyte), supporting a role for GR in regulating CaBP-9k. In ovariectomized (OVX) and ADX female mice daily treated with Dex for 3 days, duodenal CaBP-9k was expressed at the same level as in male mice. Also, no cross-activity of progesterone and Dex on their receptors was observed. Taken together, these results indicate that mouse CaBP-9k gene may be regulated by Dex in a tissue-specific manner, and reduced duodenal CaBP-9k via the GR pathway may take part in negative calcium absorption of GC-induced osteoporosis, whereas renal CaBP-9k may not be involved in the regulation of calcium homeostasis. PMID- 16219670 TI - Regulation of aldosterone production from zona glomerulosa cells by ANG II and cAMP: evidence for PKA-independent activation of CaMK by cAMP. AB - Aldosterone production in zona glomerulosa (ZG) cells of adrenal glands is regulated by various extracellular stimuli (K(+), ANG II, ACTH) that all converge on two major intracellular signaling pathways: an increase in cAMP production and calcium (Ca(2+)) mobilization. However, molecular events downstream of the increase in intracellular cAMP and Ca(2+) content are controversial and far from being completely resolved. Here, we found that Ca(2+)/calmodulin-dependent protein kinases (CaMKs) play a predominant role in the regulation of aldosterone production stimulated by ANG II, ACTH, and cAMP. The specific CaMK inhibitor KN93 strongly reduced ANG II-, ACTH-, and cAMP-stimulated aldosterone production. In in vitro kinase assays and intact cells, we could show that cAMP-induced activation of CaMK, using the adenylate cyclase activator forskolin or the cAMP analog Sp-5,6-DCI-cBIMPS (cBIMPS), was not mediated by PKA. Activation of the recently identified cAMP target protein Epac (exchange protein directly activated by cAMP) by 8-pCPT-2'-O-Me-cAMP had no effect on CaMK activity and aldosterone production. Furthermore, we provide evidence that cAMP effects in ZG cells do not involve Ca(2+) or MAPK signaling. Our results suggest that ZG cells, in addition to PKA and Epac/Rap proteins, contain other as yet unidentified cAMP mediator(s) involved in regulating CaMK activity and aldosterone secretion. PMID- 16219672 TI - Visual search improvement in hemianopic patients after audio-visual stimulation. AB - One of the most effective techniques in the rehabilitation of visual field defects is based on implementation of oculomotor strategies to compensate for visual field loss. In the present study we develop a new rehabilitation approach based on the audio-visual stimulation of the visual field. Since it has been demonstrated that audio-visual interaction in multisensory neurons can improve temporally visual perception in patients with hemianopia, the aim of the present study was to verify whether a systematic audio-visual stimulation might induce a long-lasting amelioration of visual field disorders. Eight patients with chronic visual field defects were trained to detect the presence of visual targets. During the training, the visual stimulus could be presented alone, i.e. unimodal condition, or together with an acoustic stimulus, i.e. crossmodal conditions. In the crossmodal conditions, the spatial disparity between the visual and the acoustic stimuli were systematically varied (0, 16 and 32 degrees of disparity). Furthermore, the temporal interval between the acoustic stimulus and the visual target in the crossmodal conditions was gradually reduced from 500 to 0 ms. Patients underwent the treatment for 4 h daily, over a period of nearly 2 weeks. The results showed a progressive improvement of visual detections during the training and an improvement of visual oculomotor exploration that allowed patients to efficiently compensate for the loss of vision. More interesting, there was a transfer of treatment gains to functional measures assessing visual field exploration and to daily-life activities, which was found stable at the 1 month follow-up control session. These findings are very promising with respect to the possibility of taking advantage of human multisensory capabilities to recover from unimodal sensory impairments. PMID- 16219671 TI - Autologous olfactory ensheathing cell transplantation in human spinal cord injury. AB - Olfactory ensheathing cells transplanted into the injured spinal cord in animals promote regeneration and remyelination of descending motor pathways through the site of injury and the return of motor functions. In a single-blind, Phase I clinical trial, we aimed to test the feasibility and safety of transplantation of autologous olfactory ensheathing cells into the injured spinal cord in human paraplegia. Participants were three male paraplegics, 18-55 years of age, with stable, complete thoracic injuries 6-32 months previously, with stable spinal column, no implanted prostheses, and no syrinx. Olfactory ensheathing cells were grown and purified in vitro from nasal biopsies and injected into the region of damaged spinal cord. The trial design includes a matched injury group as a control for the assessors, who are blind to treatment status. Assessments, made before transplantation and at regular intervals subsequently, include MRI, medical, neurological and psychosocial assessments, and standard American Spinal Injury Association and Functional Independence Measure assessments. One year after cell implantation, there were no medical, surgical or other complications to indicate that the procedure is unsafe. There is no evidence of spinal cord damage nor of cyst, syrinx or tumour formation. There was no neuropathic pain reported by the participants, no change in psychosocial status and no evidence of deterioration in neurological status. Participants will be followed for 3 years to confirm long-term safety and to compare neurological, functional and psychosocial outcomes with the control group. We conclude transplantation of autologous olfactory ensheathing cells into the injured spinal cord is feasible and is safe up to one year post-implantation. PMID- 16219673 TI - Magnetization transfer histograms in clinically isolated syndromes suggestive of multiple sclerosis. AB - In established multiple sclerosis, magnetization transfer ratio (MTR) histograms reveal abnormalities of normal-appearing white matter (NAWM) and grey matter (NAGM). The aim of this study was to investigate for such abnormalities in a large cohort of patients presenting with clinically isolated syndromes suggestive of multiple sclerosis. Magnetization transfer imaging was performed on 100 patients (67 women, 33 men, median age 32 years) a mean of 19 weeks (SD 3.8, range 12-33 weeks) after symptom onset with a clinically isolated syndrome and in 50 healthy controls (34 women, 16 men, median age 32.5 years). SPM99 software was used to generate segmented NAWM and NAGM MTR maps. The volumes of T2 lesions, white matter and grey matter were calculated. Eighty-one patients were followed up clinically and with conventional MRI after 3 years (n = 61) or until they developed multiple sclerosis if this occurred sooner (n = 20). Multiple regression analysis was used to investigate differences between patients and controls with age, gender and volume measures as covariates to control for potential confounding effects. The MTR histograms for both NAWM and NAGM showed a reduction in the mean (NAWM, 38.14 versus 38.33, P = 0.001; NAGM 32.29 versus 32.50, P = 0.009; units in pu) and peak location, with a left shift in the histogram. Mean NAWM and NAGM MTR were also reduced in the patients who developed clinically definite multiple sclerosis and multiple sclerosis according to the McDonald criteria but not in the 24 patients with normal T2-weighted brain magnetic resonance imaging (MRI). MTR abnormalities occur in the NAWM and NAGM at the earliest clinical stages of multiple sclerosis. PMID- 16219674 TI - Tumour necrosis factor-induced death of adult human oligodendrocytes is mediated by apoptosis inducing factor. AB - Tumour necrosis factor (TNF)-induced death of oligodendrocytes, the cell type targeted in multiple sclerosis, is mediated by TNF receptor p55 (TNFR-p55). The ligation of TNFR-p55 induces several signal transduction pathways; however, the precise mechanism involved in human oligodendrocyte (hOL) death is unknown. We defined that TNF-induced death of hOLs is non-caspase dependent, as evidenced by lack of generation of caspases 8, 1 and 3 active subunits; lack of cleavage of caspases 1 and 3 fluorogenic substrates; and lack of hOL death inhibition by the general caspase inhibitor, ZVAD.FMK. Electrophoresis of TNF-exposed hOL DNA revealed large-scale DNA fragmentation characteristic of apoptosis-inducing factor (AIF)-mediated cell death, and co-localization experiments showed that AIF translocation to the nucleus occurred upon exposure to TNF. AIF depletion by an antisense strategy prevented TNF-induced hOL death. These results indicate that TNF-induced death of hOLs is dependent on AIF, information of significance for the design strategies to protect hOLs during immune-mediated demyelination. PMID- 16219675 TI - A possible role for humoral immunity in the pathogenesis of Parkinson's disease. AB - The pathogenesis of idiopathic Parkinson's disease is unknown, but nigral degeneration and depigmentation are associated with microglial inflammation and anti-inflammatory medications appear to protect against the disease. The possibility that humoral immunity may play a role in initiating or regulating the inflammation has been suggested by experimental studies triggering dopamine cell death using a variety of transfer strategies and the observation of CD8+ T lymphocytes and complement in the nigra in Parkinson's disease. We analysed the association between degeneration and humoral immune markers in brain tissue of patients with idiopathic (n = 13) or genetic (n = 2 with alpha-synuclein and n = 1 with parkin mutations) Parkinson's disease and controls without neurological disease (n = 12) to determine the humoral immune involvement in Parkinson's disease. Formalin-fixed tissue samples from the substantia nigra and primary visual cortex for comparison were stained for alpha-synuclein, major histocompatibility complex II (HLA), immunoglobulin M (IgM), immunoglobulin G (IgG), IgG subclasses 1-4 and IgG receptors FcgammaR I-III. Antigen retrieval and both single immunoperoxidase and double immunofluorescence procedures were employed to determine the cell types involved and their pattern and semiquantitative densities. Significant dopamine neuron loss occurred in all patients with Parkinson's disease, negatively correlating with disease duration (r = -0.76, P = 0.002). Although all patients had increased inflammatory HLA immunopositive microglia, the degree of inflammation was similar throughout the disease (r = 0.08, P = 0.82). All patients with Parkinson's disease had IgG binding on dopamine neurons but not IgM binding. Lewy bodies were strongly immunolabelled with IgG. A mean 30 +/- 12% of dopamine nigral neurons were immunoreactive for IgG in Parkinson's disease with the proportion of IgG immunopositive neurons negatively correlating with the degree of cell loss in the substantia nigra (r = -0.67, P < 0.0001) and positively correlating with the number of HLA immunopositive microglia (r = 0.51, P = 0.01). Most neuronal IgG was the IgG1 subclass with some IgG3 and less IgG2 also found in the damaged substantia nigra. The high affinity activating IgG receptor, FcgammaRI, was expressed on nearby activated microglia. The low affinity activating IgG receptor, FcgammaRIII was expressed on cells morphologically resembling lymphocytes, whereas immunoreactivity for the inhibitory IgG receptor FcgammaRII was absent in all cases. This pattern of humoral immune reactivity is consistent with an immune activation of microglia leading to the targeting of dopamine nigral neurons for destruction in both idiopathic and genetic cases of Parkinson's disease. PMID- 16219676 TI - In vivo role of the phosphate groove of PDK1 defined by knockin mutation. AB - AGC kinases are mediators of signalling responses stimulated by agonists and are activated following phosphorylation at their T-loop residue by the 3 phosphoinositide-dependent protein kinase-1 (PDK1). Agonists stimulate the activation of the AGC kinases p70 ribosomal S6 kinase (S6K), p90 ribosomal S6 kinase (RSK) and serum and glucocorticoid-induced protein kinase (SGK), by inducing the phosphorylation of these enzymes at a non-catalytic regulatory site termed the hydrophobic motif. This creates a high-affinity docking site enabling PDK1 to bind and phosphorylate the T-loop of these enzymes. The site that interacts with these substrates is located on the small lobe of the catalytic domain of PDK1 and is composed of a hydrophobic groove next to a basic phosphate groove. The disruption of the hydrophobic groove ablates activation of S6K, RSK and SGK, but the role of the phosphate groove in regulating the function of PDK1 has not been explored in vivo. We generated knockin ES cells, in which both copies of the gene encoding PDK1 were altered to express a form of PDK1 that retains catalytic activity and integrity of the hydrophobic groove, but in which the phosphate groove was disrupted. The knockin ES cells were viable, mutant PDK1 was expressed at normal levels and IGF1 induced activation of protein kinase B (PKB/Akt), which is a PDK1 substrate that does not require hydrophobic motif phosphorylation to be activated. In the phosphate-groove-knockin ES cells, the activation of S6K, RSK and SGK by agonists, although markedly impaired, was not abolished. PDK1 also phosphorylates the T-loop of protein kinase C (PKC) isoforms, which stabilizes these enzymes. However, in contrast to S6K, RSK and SGK, hydrophobic motif phosphorylation of these enzymes is not thought to control their activation by PDK1. Consistent with this notion, we employed appropriate PDK1-knockin ES cells to demonstrate that the hydrophobic groove of PDK1, but not the phosphate groove, is required for the stabilization of PKC isoforms. These findings provide genetic evidence that the phosphate groove of PDK1 is required for maximal activation of isoforms of S6K, SGK and RSK, but not PKC. We also found that no live births of homozygous phosphate-groove-knockin mice are observed, indicating a key role for this regulatory motif in normal development. The knockin embryos develop to a greater extent than PDK1-knockout and hydrophobic-groove-knockin embryos, which died between E9.5-E11.5. The knockin embryos are observed until E19.5 and displayed general growth retardation and craniofacial developmental defects. PMID- 16219677 TI - CREB-binding protein regulates apoptosis and growth of HMECs grown in reconstituted ECM via laminin-5. AB - Interactions between normal mammary epithelial cells and extracellular matrix (ECM) are important for mammary gland homeostasis. Loss of interactions between ECM and normal mammary epithelial cells are thought to be an early event in mammary carcinogenesis. CREB-binding protein (CBP) is an important regulator of proliferation and apoptosis but the role of CBP in ECM signaling is poorly characterized. CBP was suppressed in basal-cytokeratin-positive HMECs (CK5/6+, CK14+, CK8-, CK18-, CK19-). Suppression of CBP resulted in loss of reconstituted ECM-mediated growth control and apoptosis and loss of laminin-5 alpha3-chain expression. Suppression of CBP in normal human mammary epithelial cells (HMECs) resulted in loss of CBP occupancy of the LAMA3A promoter and decreased LAMA3A promoter activity and laminin-5 alpha-3 chain expression. Exogenous expression of CBP in CBP-negative HMECs that have lost reconstituted ECM-mediated growth regulation and apoptosis resulted in (1) CBP occupancy of the LAMA3A promoter, (2) increased LAMA3A activity and laminin-5 alpha3-chain expression, and (3) enhancement of reconstituted ECM-mediated growth regulation and apoptosis. Similarly, suppression of laminin-5 alpha3-chain expression in HMECs resulted in loss of reconstituted ECM-mediated growth control and apoptosis. These observations suggest that loss of CBP in basal-cytokeratin-positive HMECs results in loss of reconstituted ECM-mediated growth control and apoptosis through loss of LAMA3A activity and laminin-5 alpha3-chain expression. Results in these studies may provide insight into early events in basal-type mammary carcinogenesis. PMID- 16219678 TI - Interactions between coilin and PIASy partially link Cajal bodies to PML bodies. AB - The eukaryotic nucleus contains a variety of dynamic structures, yet studies into the functional relationship of one type of subnuclear domain to another have been limited. For example, PML bodies and Cajal bodies associate, but the functional consequence of this association and the mediating factors are unknown. Here we report that an associated PML body/Cajal body can co-localize to an snRNA gene locus, with the Cajal body invariably situated between the PML body and the snRNA locus. Binding studies demonstrate that coilin (a Cajal body protein) directly interacts with PIASy (a PML body protein). Cell biological experiments using coilin and PIASy knockout cell lines demonstrate that interactions between coilin and PIASy account in part for the observed association of Cajal bodies with PML bodies. When the PIASy interaction region on coilin is deleted, the frequency of the association between Cajal bodies and PML bodies is reduced. These studies provide another example of coilin's role in the functional organization of the nucleus. PMID- 16219679 TI - The transmembrane domain is essential for the microtubular trafficking of membrane type-1 matrix metalloproteinase (MT1-MMP). AB - Membrane type-1 matrix metalloproteinase (MT1-MMP) degrades the extracellular matrix, initiates the activation pathway of soluble MMPs and regulates the functionality of cell adhesion signaling receptors, thus playing an important role in many cell functions. Intracellular transport mechanisms, currently incompletely understood, regulate the presentation of MT1-MMP at the cell surface. We have focused our efforts on identifying these mechanisms. To understand the transport of MT1-MMP across the cell, we used substitution and deletion mutants, the trafficking of which was examined using antibody uptake and Chariot delivery experiments. Our experiments have demonstrated that the microtubulin cytoskeleton and the centrosomes (the microtubulin cytoskeleton organizing centers) are essential for the trafficking and the internalization of MT1-MMP. We determined that after reaching the plasma membrane, MT1-MMP is internalized in the Rab-4-positive recycling endosomes and the Rab-11-positive pericentrosomal recycling endosomes. The microtubular trafficking causes the protease to accumulate in the pericentrosomal region of the cell. We believe that the presence of the transmembrane domain is required for the microtubular vesicular trafficking of MT1-MMP because the soluble mutants are not presented at the cell surface and they are not delivered to the centrosomes. The observed transport mechanisms provide a vehicle for the intracellular targets and, accordingly, for an intracellular cleavage function of MT1-MMP in malignant cells, which routinely overexpress this protease. PMID- 16219680 TI - Charged bipolar suramin derivatives induce aggregation of the prion protein at the cell surface and inhibit PrPSc replication. AB - The conversion of the cellular prion protein (PrPc) into a pathogenic isoform (PrP(Sc)) is one of the underlying events in the pathogenesis of the fatal transmissible spongiform encephalopathies (TSEs). Numerous compounds have been described to inhibit prion replication and PrP(Sc) accumulation in cell culture. Among these, the drug suramin induces aggregation and re-targeting of PrPc to endocytic compartments. Plasma membrane and sites of conversion into PrP(Sc) are thereby bypassed. In the present study, a library of suramin analogues was tested as a potential class of new anti-prion compounds and the molecular mechanisms underlying these effects were analysed. Treatment of prion-infected neuroblastoma cells with compounds containing symmetrical aromatic sulfonic acid substitutions inhibited de novo synthesis of PrP(Sc) and induced aggregation and reduction of the half-life of PrPc without downregulating PrPc cell surface expression. Half molecule compounds lacking the symmetrical bipolar structure or the anionic groups had no effect on PrP(Sc) synthesis or PrPc solubility. Cell surface expression of PrPc was necessary for the activity of effective compounds. Suramin derivatives did not induce aggregation of PrPc when transport along the secretory pathway was compromised, suggesting that their effects occur at a post trans Golgi network (TGN) site, possibly close to the compartment of conversion into PrP(Sc). In vitro studies with recombinant PrP demonstrated that the inhibitory effect correlated with direct binding to PrP and induction of insoluble PrP aggregates. Our data reveal an anti-prion effect that differs from those characterising other sulphated polyanions and is dependent on the presence of the symmetrical anionic structure of these molecules. PMID- 16219681 TI - The NF-kappaB pathway. PMID- 16219682 TI - Usher I syndrome: unravelling the mechanisms that underlie the cohesion of the growing hair bundle in inner ear sensory cells. AB - Defects in myosin VIIa, the PDZ-domain-containing protein harmonin, cadherin 23 and protocadherin 15 (two cadherins with large extracellular regions), and the putative scaffolding protein Sans underlie five genetic forms of Usher syndrome type I (USH1), the most frequent cause of hereditary deafness-blindness in humans. All USH1 proteins are localised within growing stereocilia and/or the kinocilium that make up the developing auditory hair bundle, the mechanosensitive structure receptive to sound stimulation. Cadherin 23 has been shown to be a component of fibrous links interconnecting the growing stereocilia as well as the kinocilium and the nearest tall stereocilia. A similar function is anticipated for protocadherin 15. Multiple direct interactions between USH1 proteins have been demonstrated. In particular, harmonin b can bind to the cytoplasmic regions of cadherin 23 and protocadherin 15, and to F-actin, and thus probably anchors these cadherins to the actin filaments filling the stereocilia. Myosin VIIa and Sans are both involved in the sorting and/or targeting of harmonin b to the stereocilia. Together, this suggests that the disorganisation of the hair bundles observed in mice mutants lacking orthologues of USH1 proteins may result from a defect of hair-bundle-link-mediated adhesion forces. Moreover, several recent evidences suggest that some genes defective in Usher type II syndrome also encode interstereocilia links, thus bridging the pathogenic pathways of USH1 and USH2 hearing impairment. Additional functions of USH1 proteins in the inner ear and the retina are evident from other phenotypic abnormalities observed in these mice. In particular, myosin VIIa could act at the interface between microtubule- and actin-based transport. PMID- 16219683 TI - Sphingosine 1-phosphate and ceramide 1-phosphate: expanding roles in cell signaling. AB - The phosphorylated sphingolipid metabolites sphingosine 1-phosphate (S1P) and ceramide 1-phosphate (C1P) have emerged as potent bioactive agents. Recent studies have begun to define new biological functions for these lipids. Generated by sphingosine kinases and ceramide kinase, they control numerous aspects of cell physiology, including cell survival and mammalian inflammatory responses. Interestingly, S1P is involved in cyclooxygenase-2 induction and C1P is required for the activation and translocation of cPLA2. This suggests that these two sphingolipid metabolites may act in concert to regulate production of eicosanoids, important inflammatory mediators. Whereas S1P functions mainly via G protein-coupled receptors, C1P appears to bind directly to targets such as cPLA2 and protein phosphatase 1/2A. S1P probably also has intracellular targets, and in plants it appears to directly regulate the G protein alpha subunit GPA1. PMID- 16219684 TI - Yeast Mon2p is a highly conserved protein that functions in the cytoplasm-to vacuole transport pathway and is required for Golgi homeostasis. AB - Although the small Arf-like GTPases Arl1-3 are highly conserved eukaryotic proteins, they remain relatively poorly characterized. The yeast and mammalian Arl1 proteins bind to the Golgi complex, where they recruit specific structural proteins such as Golgins. Yeast Arl1p directly interacts with Mon2p/Ysl2p, a protein that displays some sequence homology to the large Sec7 guanine exchange factors (GEFs) of Arf1. Mon2p also binds the putative aminophospholipid translocase (APT) Neo1p, which performs essential function(s) in membrane trafficking. Our detailed analysis reveals that Mon2p contains six distinct amino acid regions (A to F) that are conserved in several other uncharacterized homologs in higher eukaryotes. As the conserved A, E and F domains are unique to these homologues, they represent the signature of a new protein family. To investigate the role of these domains, we made a series of N- and C-terminal deletions of Mon2p. Although fluorescence and biochemical studies showed that the B and C domains (also present in the large Sec7 GEFs) predominantly mediate interaction with Golgi/endosomal membranes, growth complementation studies revealed that the C-terminal F domain is essential for the activity of Mon2p, indicating that Mon2p might also function independently of Arl1p. We provide evidence that Mon2p is required for efficient recycling from endosomes to the late Golgi. Intriguingly, although transport of CPY to the vacuole was nearly normal in the Deltamon2 strain, we found the constitutive delivery of Aminopeptidase 1 from the cytosol to the vacuole to be almost completely blocked. Finally, we show that Mon2p exhibits genetic and physical interactions with Dop1p, a protein with a putative function in cell polarity. We propose that Mon2p is a scaffold protein with novel conserved domains, and is involved in multiple aspects of endomembrane trafficking. PMID- 16219685 TI - E-cadherin endocytosis regulates the activity of Rap1: a traffic light GTPase at the crossroads between cadherin and integrin function. AB - The coordinate modulation of cadherin and integrin functions plays an essential role in fundamental physiological and pathological processes, including morphogenesis and cancer. However, the molecular mechanisms underlying the functional crosstalk between cadherins and integrins are still elusive. Here, we demonstrate that the small GTPase Rap1, a crucial regulator of the inside-out activation of integrins, is a target for E-cadherin-mediated outside-in signaling. In particular, we show that a strong activation of Rap1 occurs upon adherens junction disassembly that is triggered by E-cadherin internalization and trafficking along the endocytic pathway. By contrast, Rap1 activity is not influenced by integrin outside-in signaling. Furthermore, we demonstrate that the E-cadherin endocytosis-dependent activation of Rap1 is associated with and controlled by an increased Src kinase activity, and is paralleled by the colocalization of Rap1 and E-cadherin at the perinuclear Rab11-positive recycling endosome compartment, and the association of Rap1 with a subset of E-cadherin catenin complexes that does not contain p120ctn. Conversely, Rap1 activity is suppressed by the formation of E-cadherin-dependent cell-cell junctions as well as by agents that inhibit either Src activity or E-cadherin internalization and intracellular trafficking. Finally, we demonstrate that the E-cadherin endocytosis-dependent activation of Rap1 is associated with and is required for the formation of integrin-based focal adhesions. Our findings provide the first evidence of an E-cadherin-modulated endosomal signaling pathway involving Rap1, and suggest that cadherins may have a novel modulatory role in integrin adhesive functions by fine-tuning Rap1 activation. PMID- 16219686 TI - Interaction between secretogranin III and carboxypeptidase E facilitates prohormone sorting within secretory granules. AB - Secretogranin III (SgIII) and carboxypeptidase E (CPE) bind specifically to cholesterol-rich secretory granule (SG) membranes. We previously showed that SgIII binds chromogranin A (CgA) and targets CgA to the SGs in endocrine cells. We investigated the binding of SgIII and CPE because they frequently localize close to the periphery of SGs, and they bind each other in mouse corticotrope derived AtT-20 cells. In Cpe fat mouse corticotropes, which have defective CPE, proopiomelanocortin (POMC)-derived adrenocorticotrophin hormone (ACTH)-containing peptides were distributed over the entire surface of the SGs, and displayed a regulated secretion by secretagogues. The Cpe fat pituitary exhibited elevated levels of SgIII and CgA, which suggests that they compensate for a sorting function of CPE for POMC and its intermediates to ACTH. Indeed, both SgIII and CgA were able to bind POMC-derived intermediates. In a competitive pull-down assay, excessive SgIII led to a decrease in CPE-bound POMC-derived intermediate molecules, and SgIII pulled-down by anti-ACTH antibody increased proportionately. We suggest that SgIII and CPE form the separate functional sorting complex by anchoring to cholesterol-rich SG membranes, and POMC-derived peptides are transferred from CPE to SgIII, and subsequently to CgA. PMID- 16219688 TI - Evidence that satellite cell decrement contributes to preferential decline in nuclear number from large fibres during murine age-related muscle atrophy. AB - Skeletal muscle fibres are multinucleate syncitial cells that change size during adult life depending on functional demand. The relative contribution of change in nuclear number and/or cell growth to fibre size change is unclear. We report that nuclei/unit length decreases in larger fibres during skeletal muscle ageing. This leads to an increased size of nuclear domain (quantity of cytoplasm/number of nuclei within that cytoplasm). Initially, larger fibres have more satellite cells than small fibres, but this advantage is lost as satellite cells decline with age. These changes are accompanied by an overall decline in fibre size, returning domain size to the normal range. Exacerbated loss of fibre nuclei per unit length during ageing of myoD-null mice provides the first experimental support for the hypothesis that a satellite cell defect causes inadequate nuclear replacement. We propose a model in which a decline in satellite cell function and/or number during ageing leads to a loss of nuclei from large fibres and an associated domain size increase that triggers cytoplasmic atrophy through the normal cell size-regulating machinery. PMID- 16219687 TI - Actin and non-muscle myosin II facilitate apical exocytosis of tear proteins in rabbit lacrimal acinar epithelial cells. AB - The acinar epithelial cells of the lacrimal gland exocytose the contents of mature secretory vesicles containing tear proteins at their apical membranes in response to secretagogues. Here we use time-lapse confocal fluorescence microscopy and fluorescence recovery after photobleaching to investigate the changes in actin filaments located beneath the apical membrane during exocytosis evoked by the muscarinic agonist, carbachol (100 microM). Time-lapse confocal fluorescence microscopy of apical actin filaments in reconstituted rabbit lacrimal acini transduced with replication-deficient adenovirus containing GFP actin revealed a relatively quiescent apical actin array in resting acini. Carbachol markedly increased apical actin filament turnover and also promoted transient actin assembly around apparent fusion intermediates. Fluorescence recovery after photobleaching measurements revealed significant (P< or =0.05) increases and decreases, respectively, in mobile fraction (Mf) and turnover times (t1/2) for apical actin filaments in carbachol-stimulated acini relative to untreated acini. The myosin inhibitors, 2,3-butanedione monoxime (BDM, 10 mM, 15 minutes) and ML-7 (40 microM, 15 minutes), significantly decreased carbachol stimulated secretion of bulk protein and the exogenous secretory vesicle marker, syncollin-GFP; these agents also promoted accumulation of actin-coated structures which were enriched, in transduced acini, in syncollin-GFP, confirming their identity as fusion intermediates. Actin-coated fusion intermediates were sized consistent with incorporation of multiple rather than single secretory vesicles; moreover, BDM and ML-7 caused a shift towards formation of multiple secretory vesicle aggregates while significantly increasing the diameter of actin-coated fusion intermediates. Our findings suggest that the increased turnover of apical actin filaments and the interaction of actin with non-muscle myosin II assembled around aggregates of secretory vesicles facilitate exocytosis in lacrimal acinar epithelial cells. PMID- 16219689 TI - Myosin Ib modulates the morphology and the protein transport within multi vesicular sorting endosomes. AB - Members of at least four classes of myosin (I, II, V and VI) have been implicated in the dynamics of a large variety of organelles. Despite their common motor domain structure, some of these myosins, however, are non processive and cannot move organelles along the actin tracks. Here, we demonstrate in the human pigmented MNT-1 cell line that, (1) the overexpression of one of these myosins, myosin 1b, or the addition of cytochalasin D affects the morphology of the sorting multivesicular endosomes; (2) the overexpression of myosin 1b delays the processing of Pmel17 (the product of murine silver locus also named GP100), which occurs in these multivesicular endosomes; (3) myosin 1b associated with endosomes coimmunoprecipitates with Pmel17. All together, these observations suggest that myosin 1b controls the traffic of protein cargo in multivesicular endosomes most probably through its ability to modulate with actin the morphology of these sorting endosomes. PMID- 16219690 TI - Cholesterol facilitates the native mechanism of Ca2+-triggered membrane fusion. AB - The process of regulated exocytosis is defined by the Ca2+-triggered fusion of two apposed membranes, enabling the release of vesicular contents. This fusion step involves a number of energetically complex steps and requires both protein and lipid membrane components. The role of cholesterol has been investigated using isolated release-ready native cortical secretory vesicles to analyze the Ca2+-triggered fusion step of exocytosis. Cholesterol is a major component of vesicle membranes and we show here that selective removal from membranes, selective sequestering within membranes, or enzymatic modification causes a significant inhibition of the extent, Ca2+ sensitivity and kinetics of fusion. Depending upon the amount incorporated, addition of exogenous cholesterol to cholesterol-depleted membranes consistently recovers the extent, but not the Ca2+ sensitivity or kinetics of fusion. Membrane components of comparable negative curvature selectively recover the ability to fuse, but are unable to recover the kinetics and Ca2+ sensitivity of vesicle fusion. This indicates at least two specific positive roles for cholesterol in the process of membrane fusion: as a local membrane organizer contributing to the efficiency of fusion, and, by virtue of its intrinsic negative curvature, as a specific molecule working in concert with protein factors to facilitate the minimal molecular machinery for fast Ca2+ triggered fusion. PMID- 16219691 TI - Regulation of paxillin family members during epithelial-mesenchymal transformation: a putative role for paxillin delta. AB - Epithelial-mesenchymal transformation (EMT) and the resulting induction of cell motility are essential components of tissue remodeling during embryonic development and wound repair, as well as tumor progression to an invasive metastatic phenotype. Paxillin, a multi-domain adaptor and phosphoprotein has previously been implicated in integrin signaling and cell motility. In this report we characterize a novel paxillin gene product, paxillin delta, generated from an evolutionarily conserved internal translation initiation site within the full-length paxillin mRNA. Paxillin delta, which lacks the key phosphorylation sites Y31 and Y118 as well as the ILK and actopaxin binding LD1 motif, exhibits a restricted distribution to epithelial cell types and is downregulated during TGF beta1-induced EMT of normal murine mammary gland (NMuMG) epithelial cells. Interestingly, Hic-5, a paxillin superfamily member, exhibits a reciprocal protein expression profile to paxillin delta. In addition, paxillin delta expression is maintained following NMuMG differentiation in a 3D collagen I gel while other focal adhesion components are downregulated. Paxillin delta protein expression coincided with reduced paxillin tyrosine phosphorylation in NMuMG cells and paxillin delta overexpression in CHO.K1 cells inhibited adhesion mediated tyrosine phosphorylation of paxillin. Forced expression of paxillin delta in NMuMG cells suppressed cell migration whereas Hic-5 overexpression stimulated motility. Together our data support a role for paxillin delta as a naturally occurring functional antagonist of paxillin signaling potentially through suppression of a Crk-mediated pathway during processes associated with cell migration. PMID- 16219692 TI - Evidence for the involvement of PECAM-1 in a receptor mediated signal transduction pathway regulating capacitation-associated tyrosine phosphorylation in human spermatozoa. AB - Mammalian spermatozoa must become ;capacitated' in the female reproductive tract before they gain the ability to fertilize the oocyte. The attainment of a capacitated state has been correlated with a number of biochemical changes, the most notable of which is a dramatic increase in the tyrosine phosphorylation status of these cells. Despite its biological importance, the mechanisms responsible for initiating this tyrosine phosphorylation cascade in vivo are unknown. Here, we report that this signalling pathway can be elicited in a rapid, dose-dependent and lectin-specific manner by wheat germ agglutinin (WGA), but none of 18 other lectins assessed. This response was abrogated by prior enzymatic cleavage of either sialic acid or GlcNAc residues from the sperm surface and by treatment with a range of pharmacological inhibitors directed against protein kinase A, protein tyrosine kinases and intermediates including Src. Proteomic analysis of the WGA-binding sites on the sperm surface identified the putative cognate receptor as platelet cell adhesion molecule 1 (PECAM-1/CD31). This conclusion was supported by the following evidence: (i) anti-PECAM-1 antibodies identified a molecule of the correct molecular mass in human spermatozoa, (ii) PECAM-1 could be isolated from a pool of sperm surface proteins using WGA immobilized on a solid phase support, (iii) PECAM-1 and WGA co-localized to the sperm surface and (iv) anti-PECAM-1 antibodies could completely block the ability of WGA to stimulate tyrosine phosphorylation in these cells. Collectively, these data provide the first evidence that a receptor-mediated signal transduction pathway triggers human sperm capacitation and identifies PECAM-1 as the probable initiator of this second messenger cascade. PMID- 16219693 TI - Selective cellular effects of overexpressed pleckstrin-homology domains that recognize PtdIns(3,4,5)P3 suggest their interaction with protein binding partners. AB - Several pleckstrin-homology (PH) domains with the ability to bind phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P3, PIP3] were expressed as green fluorescent protein (GFP) fusion proteins to determine their effects on various cellular responses known to be activated by PIP3. These proteins comprised the PH domains of Akt, ARNO, Btk or GRP1, and were found to show growth factor-stimulated and wortmannin-sensitive translocation from the cytosol to the plasma membrane in several cell types, indicating their ability to recognize PIP3. Remarkably, although overexpressed Akt-PH-GFP and Btk-PH-GFP were quite potent in antagonizing the PIP3-mediated activation of the Akt protein kinase, such inhibition was not observed with the other PH domains. By contrast, expression of the PH domains of GRP1 and ARNO, but not of Akt or Btk, inhibited the attachment and spreading of freshly seeded cells to culture dishes. Activation of PLCgamma by epidermal growth factor (EGF) was attenuated by the PH domains of GRP1, ARNO and Akt, but was significantly enhanced by the Btk PH domain. By following the kinetics of expression of the various GFP-fused PH domains for several days, only the PH domain of Akt showed a lipid-binding dependent self-elimination, consistent with its interference with the anti apoptotic Akt signaling pathway. Mutations of selective residues that do not directly participate in PIP3 binding in the GRP1-PH and Akt-PH domain were able to reduce the dominant-negative effects of these constructs yet retain their lipid binding. These data suggest that interaction with and sequestration of PIP3 may not be the sole mechanism by which PH domains interfere with cellular responses and that their interaction with other membrane components, most probably with proteins, allows a more specific participation in the regulation of specific signaling pathways. PMID- 16219694 TI - Silencing Cenp-F weakens centromeric cohesion, prevents chromosome alignment and activates the spindle checkpoint. AB - Cenp-F is an unusual kinetochore protein in that it localizes to the nuclear matrix in interphase and the nuclear envelope at the G2/M transition; it is farnesylated and rapidly degraded after mitosis. We have recently shown that farnesylation of Cenp-F is required for G2/M progression, its localization to kinetochores, and its degradation. However, the role Cenp-F plays in mitosis has remained enigmatic. Here we show that, following repression of Cenp-F by RNA interference (RNAi), the processes of metaphase chromosome alignment, anaphase chromosome segregation and cytokinesis all fail. Although kinetochores attach to microtubules in Cenp-F-deficient cells, the oscillatory movements that normally occur following K-fibre formation are severely dampened. Consistently, inter kinetochore distances are reduced. In addition, merotelic associations are observed, suggesting that whereas kinetochores can attach microtubules in the absence of Cenp-F, resolving inappropriate interactions is inhibited. Repression of Cenp-F does not appear to compromise the spindle checkpoint. Rather, the chromosome alignment defect induced by Cenp-F RNA interference is accompanied by a prolonged mitosis, indicating checkpoint activation. Indeed, the prolonged mitosis induced by Cenp-F RNAi is dependent on the spindle checkpoint kinase BubR1. Surprisingly, chromosomes in Cenp-F-deficient cells frequently show a premature loss of chromatid cohesion. Thus, in addition to regulating kinetochore microtubule interactions, Cenp-F might be required to protect centromeric cohesion prior to anaphase commitment. Intriguingly, whereas most of the sister less kinetochores cluster near the spindle poles, some align at the spindle equator, possibly through merotelic or lateral orientations. PMID- 16219695 TI - TGFbeta-induced downregulation of E-cadherin-based cell-cell adhesion depends on PI3-kinase and PTEN. AB - Transforming growth factor beta (TGFbeta) has profound growth-suppressive effects on normal epithelial cells, but supports metastasis formation in many tumour types. In most epithelial tumour cells TGFbeta(1) treatment results in epithelial dedifferentiation with reduced cell aggregation and enhanced cellular migration. Here we show that the epithelial dedifferentiation, accompanied by dissociation of the E-cadherin adhesion complex, induced by TGFbeta(1) depended on phosphatidylinositol 3-kinase (PI3-kinase) and the phosphatase PTEN as analysed in PANC-1 and Smad4-deficient BxPC-3 pancreatic carcinoma cells. TGFbeta(1) treatment enhanced tyrosine phosphorylation of alpha- and beta-catenin, which resulted in dissociation of the E-cadherin/catenin complex from the actin cytoskeleton and reduced cell-cell adhesion. The PI3-kinase and PTEN were found associated with the E-cadherin/catenin complex via beta-catenin. TGFbeta(1) treatment reduced the amount of PTEN bound to beta-catenin and markedly increased the tyrosine phosphorylation of beta-catenin. By contrast, forced expression of PTEN clearly reduced the TGFbeta(1)-induced phosphorylation of beta-catenin. The TGFbeta(1)-induced beta-catenin phosphorylation was also dependent on PI3-kinase and Ras activity. The described effects of TGFbeta(1) were independent of Smad4, which is homozygous deleted in BxPC-3 cells. Collectively, these data show that the TGFbeta(1)-induced destabilisation of E-cadherin-mediated cell-cell adhesion involves phosphorylation of beta-catenin, which is regulated by E-cadherin adhesion complex-associated PI3-kinase and PTEN. PMID- 16219696 TI - A simulation-based approach to forecasting the next great San Francisco earthquake. AB - In 1906 the great San Francisco earthquake and fire destroyed much of the city. As we approach the 100-year anniversary of that event, a critical concern is the hazard posed by another such earthquake. In this article, we examine the assumptions presently used to compute the probability of occurrence of these earthquakes. We also present the results of a numerical simulation of interacting faults on the San Andreas system. Called Virtual California, this simulation can be used to compute the times, locations, and magnitudes of simulated earthquakes on the San Andreas fault in the vicinity of San Francisco. Of particular importance are results for the statistical distribution of recurrence times between great earthquakes, results that are difficult or impossible to obtain from a purely field-based approach. PMID- 16219697 TI - Method for targeting protein destruction by using a ubiquitin-independent, proteasome-mediated degradation pathway. AB - With the euchromatic portion of several mammalian genomes now sequenced, emphasis has turned to ascertaining the functions of gene products. A method for targeting destruction of selected proteins in mammalian cells is described, based on the ubiquitin-independent mechanism by which ornithine decarboxylase (ODC) is degraded by the 26S proteasome in collaboration with antizyme (AZ). We show that expressing whole proteins, protein domains, or peptide ligands fused to the N terminus of ODC promotes proteasome-dependent degradation of these chimeric fusion proteins and their interacting cellular target proteins. Moreover, the degradation of the interacting (targeted) protein depends on coexpression of AZ in about half of cases, providing an inducible switch for triggering the degradation process. By using 12 pairs of interacting proteins for testing, direct comparisons with several alternative strategies for achieving targeted protein destruction based on the concept of induced ubiquitination revealed advantages of the ODC/AZ system, which does not require posttranslational attachment of ubiquitin to target proteins. As proof of concept, the ODC/AZ system was used to ablate expression of specific endogenous proteins (e.g., TRAF6; Rb), and was shown to create the expected lesions in cellular pathways that require these proteins. Altogether, these findings reveal a strategy for achieving targeted destruction of cellular proteins, thus providing an additional tool for revealing the cellular phenotypes of gene products. PMID- 16219698 TI - HIV Gag protein conjugated to a Toll-like receptor 7/8 agonist improves the magnitude and quality of Th1 and CD8+ T cell responses in nonhuman primates. AB - Induction and maintenance of antibody and T cell responses will be critical for developing a successful vaccine against HIV. A rational approach for generating such responses is to design vaccines or adjuvants that have the capacity to activate specific antigen-presenting cells. In this regard, dendritic cells (DCs) are the most potent antigen-presenting cells for generating primary T cell responses. Here, we report that Toll-like receptor (TLR) agonists and ligands that activate DCs in vitro influence the magnitude and quality of the cellular immune response in nonhuman primates (NHPs) when administered with HIV Gag protein. NHPs immunized with HIV Gag protein and a TLR7/8 agonist or a TLR9 ligand [CpG oligodeoxynucleotides (CpG ODN)] had significantly increased Gag specific T helper 1 and antibody responses, compared with animals immunized with HIV Gag protein alone. Importantly, conjugating the HIV Gag protein to the TLR7/8 agonist (Gag-TLR7/8 conjugate) dramatically enhanced the magnitude and altered the quality of the T helper 1 response, compared with animals immunized with HIV Gag protein and the TLR7/8 agonist or CpG ODN. Furthermore, immunization with the Gag-TLR7/8 conjugate vaccine elicited Gag-specific CD8+ T responses. Collectively, our results show that conjugating HIV Gag protein to a TLR7/8 agonist is an effective way to elicit broad-based adaptive immunity in NHPs. This type of vaccine formulation should have utility in preventive or therapeutic vaccines in which humoral and cellular immunity is required. PMID- 16219699 TI - Antibody vs. HIV in a clash of evolutionary titans. AB - HIV has evolved many strategies to avoid neutralizing antibody responses, particularly to conserved regions on the external glycoprotein spikes of the virus. Nevertheless, a small number of antibodies have been evolved by the human immune system to recognize conserved parts of the glycoproteins, and therefore, have broadly neutralizing cross-strain activities. These antibodies constitute important tools in the quest to design immunogens that can elicit broadly neutralizing antibodies in humans and hence contribute to an effective HIV vaccine. Crystallographic analyses of the antibodies, in many cases in an antigen complexed form, have revealed novel and, in some instances, remarkable structural adaptations to attain virus recognition. Antibodies, like HIV, can evolve relatively rapidly through mutation and selection. It seems that the structures of these broadly neutralizing antibodies bear witness to a heroic struggle between two titans of rapid evolution. PMID- 16219700 TI - Single-particle tracking of murine polyoma virus-like particles on live cells and artificial membranes. AB - The lateral mobility of individual murine polyoma virus-like particles (VLPs) bound to live cells and artificial lipid bilayers was studied by single fluorescent particle tracking using total internal reflection fluorescence microscopy. The particle trajectories were analyzed in terms of diffusion rates and modes of motion as described by the moment scaling spectrum. Although VLPs bound to their ganglioside receptor in lipid bilayers exhibited only free diffusion, analysis of trajectories on live 3T6 mouse fibroblasts revealed three distinct modes of mobility: rapid random motion, confined movement in small zones (30-60 nm in diameter), and confined movement in zones with a slow drift. After binding to the cell surface, particles typically underwent free diffusion for 5 10 s, and then they were confined in an actin filament-dependent manner without involvement of clathrin-coated pits or caveolae. Depletion of cholesterol dramatically reduced mobility of VLPs independently of actin, whereas inhibition of tyrosine kinases had no effect on confinement. The results suggested that clustering of ganglioside molecules by the multivalent VLPs induced transmembrane coupling that led to confinement of the virus/receptor complex by cortical actin filaments. PMID- 16219701 TI - Biological imaging by soft x-ray diffraction microscopy. AB - We have used the method of x-ray diffraction microscopy to image the complex valued exit wave of an intact and unstained yeast cell. The images of the freeze dried cell, obtained by using 750-eV x-rays from different angular orientations, portray several of the cell's major internal components to 30-nm resolution. The good agreement among the independently recovered structures demonstrates the accuracy of the imaging technique. To obtain the best possible reconstructions, we have implemented procedures for handling noisy and incomplete diffraction data, and we propose a method for determining the reconstructed resolution. This work represents a previously uncharacterized application of x-ray diffraction microscopy to a specimen of this complexity and provides confidence in the feasibility of the ultimate goal of imaging biological specimens at 10-nm resolution in three dimensions. PMID- 16219702 TI - Statin therapy might be beneficial for patients with ankylosing spondylitis. PMID- 16219703 TI - Combining information obtained from magnetic resonance imaging and conventional radiographs to detect sacroiliitis in patients with recent onset inflammatory back pain. AB - OBJECTIVE: To compare the contribution of changes on magnetic resonance imaging (MRI) and conventional radiography (CR) in the sacroiliac joints of patients with recent onset inflammatory back pain (IBP) in making an early diagnosis of spondyloarthritides. METHODS: The study involved 68 patients with IBP (38% male; mean (SD) age, 34.9 (10.3) years) with symptom duration less than two years. Coronal MRI of the sacroiliac joints was scored for inflammation and structural changes, and pelvic radiographs were scored by the modified New York (mNY) grading. Agreement between MRI and CR was analysed by cross tabulation per sacroiliac joint and per patient. RESULTS: A structural change was detected in 20 sacroiliac joints by MRI and in 37 by CR. Inflammation was detected in 36 sacroiliac joints by MRI, and 22 of these showed radiographic sacroiliitis. Fourteen patients fulfilled the mNY criteria based on CR. Classification according to the modified New York criteria would be justified for eight patients if it was based on MRI for structural changes only, for 14 if it was based on structural changes on CR, for 14 (partly) different patients if it was based on inflammation on MRI only, for 16 if it was based on inflammation and structural changes on MRI, for 19 if it was based on inflammation on CR combined with MRI, and for (the same) 19 if it was based on inflammation and structural damage on CR combined with MRI. CONCLUSIONS: CR can detect structural changes in SI joints with higher sensitivity than MRI. However, inflammation on MRI can be found in a substantial proportion of patients with IBP but normal radiographs. Assessment of structural changes by CR followed by assessment of inflammation on MRI in patients with negative findings gives the highest returns for detecting involvement of the SI joints by imaging in patients with recent onset IBP. PMID- 16219704 TI - Determinants of hyperkyphosis in patients with ankylosing spondylitis. AB - OBJECTIVE: To determine clinical and radiographic determinants of hyperkyphosis in patients with ankylosing spondylitis. METHODS: Spinal hyperkyphosis was assessed by occiput to wall distance (OWD) in 135 patients participating in the OASIS cohort and defined as OWD >0. Disease activity was assessed by the Bath ankylosing spondylitis disease activity index (BASDAI). Wedging of the vertebrae was calculated as the Ha/Hp ratio. Structural damage of the spine was assessed by the modified Stoke ankylosing spondylitis spine score (mSASSS). Hip involvement was assessed by the Bath ankylosing spondylitis radiology index (BASRI) and defined as a score >2. Data were analysed by multiple regression analysis on van der Waerden-normal OWD values, with mean Ha/Hp ratio, mSASSS, hip involvement, and BASDAI as explanatory variables, and age, sex, and disease duration after diagnosis as covariates. RESULTS: 61 patients (45.2%) had an OWD >0 cm. Of these, 81% were male, v 57% in the group with normal OWD (p<0.001). Forty two patients had wedged thoracic vertebrae, and 27 of these (44%) had an increased OWD, compared with 15 of 74 with a normal OWD (20%) (p = 0.005). OWD was correlated with mean wedging of the thoracic spine (r = -0.45, p = 0.01), mSASSS (r = 0.56, p = 0.01), and hip involvement (r = 0.2, p = 0.05). Multivariate analysis showed that mSASSS (standardised beta (stbeta) = 0.52; p<0.001), wedging of the thoracic spine (stbeta = -0.28; p = 0.01), and BASDAI (stbeta = 0.15; p = 0.05) were independent determinants of OWD. CONCLUSIONS: Radiological damage of the cervical and lumbar spine, thoracic wedging, and disease activity are determinants of hyperkyphosis in patients with ankylosing spondylitis. These findings could be important in determining treatment goals in this disease. PMID- 16219705 TI - Low grade radiographic sacroiliitis as prognostic factor in patients with undifferentiated spondyloarthritis fulfilling diagnostic criteria for ankylosing spondylitis throughout follow up. AB - OBJECTIVE: To determine the rate and factors associated with ankylosing spondylitis in a cohort of patients with undifferentiated spondyloarthritides (SpA). METHODS: 62 consecutive patients with undifferentiated SpA seen between 1998 and 1999 underwent clinical and imaging evaluations throughout follow up. The main outcome measure was a diagnosis of ankylosing spondylitis. RESULTS: 50 patients with peripheral arthritis (n = 35) and inflammatory back pain (n = 24) (26 male; mean (SD) age at onset, 20.4 (8.8) years; disease duration 5.4 (5.7) years) were followed up for 3-5 years. At baseline, >90% of patients had axial and peripheral disease, while 38% had radiographic sacroiliitis below the cut off level for a diagnosis of ankylosing spondylitis (BASDAI 3.9, BASFI 2.9). At the most recent evaluation, 21 patients (42%) had ankylosing spondylitis. Two factors were associated with a diagnosis of ankylosing spondylitis in multivariate analysis: radiographic sacroiliitis grade <2 bilateral, or grade <3 unilateral (odds ratio (OR) = 11.18 (95% confidence interval, 2.59 to 48.16), p = 0.001), particularly grade 1 bilateral (OR = 12.58 (1.33 to 119.09), p = 0.027), and previous uveitis (OR = 19.25 (1.72 to 214.39), p = 0.001). Acute phase reactant levels, juvenile onset, and HLA-B27 showed a trend to linkage with ankylosing spondylitis (NS). CONCLUSIONS: Low grade radiographic sacroiliitis is a prognostic factor for ankylosing spondylitis in patients originally classified as having undifferentiated SpA. Low grade radiographic sacroiliitis should be regarded as indicative of early ankylosing spondylitis in patients with undifferentiated SpA. PMID- 16219706 TI - MRI bone oedema predicts eight year tendon function at the wrist but not the requirement for orthopaedic surgery in rheumatoid arthritis. AB - OBJECTIVE: To investigate the role of early magnetic resonance imaging (MRI) of the wrist in predicting functional outcome in rheumatoid arthritis. METHODS: MRI scans of the dominant wrist were scored for synovitis, tendon inflammation, bone oedema, and erosion at first presentation (n = 42), at 1 year (n = 42), and at 6 years (n = 31). At 8 years, clinical reassessment (n = 28) was undertaken. Tendon function was graded 0-3 for movement, tendon sheath swelling, and pain on resistance at nine flexor and extensor tendons of the hand. Hand function was also assessed using the Sollerman grip test. The requirement for joint or tendon surgery by 8 years was determined by telephone survey in 39 of the original 42 patients. RESULTS: At 8 years, tendon function was highly correlated with hand function (Sollerman score, R = -0.51, p = 0.005) and global function (health assessment questionnaire score, R = 0.53, p = 0.004). Using a model incorporating baseline and 1 year MRI scores, the MRI bone oedema score was strongly predictive of tendon function at 8 years (chi(2)(2) = 15.3, p = 0.0005), as was the MRI bone erosion score (chi(2)(2) = 9.23, p = 0.01). Hand function was also predicted by the baseline MRI erosion score (p = 0.02). MRI variables did not predict the requirement for surgery, but patients who had surgery were more likely to show progression of MRI bone erosion scores between baseline and 1 year (p = 0.008). CONCLUSIONS: Extensive MRI bone oedema and erosions at the wrist in early rheumatoid arthritis predict tendon dysfunction and impaired hand function in the medium term but not the requirement for joint or tendon surgery. PMID- 16219707 TI - Diagnosis and management of adult onset Still's disease. AB - BACKGROUND: Adult onset Still's disease (AOSD) is a rare systemic inflammatory disorder of unknown aetiology that is responsible for a significant proportion of cases of fever of unknown origin and can also have serious musculoskeletal sequelae. OBJECTIVE: To assess and synthesise the evidence for optimal diagnosis and management of AOSD. METHODS: The key terms, adult onset Still's disease, AOSD, adult Still's disease, ASD, Still's disease were used to search Medline (1966-2005) and PubMed (1966-2005) for all available articles in the English language. Clinically relevant articles were subsequently selected. Bibliographies, textbooks, and websites of recent rheumatology conferences were also assessed. RESULTS: Data on diagnosis and treatment of AOSD are limited in the medical literature and consist mainly of case reports, small series, and modest scale retrospective studies. Diagnosis is clinical and requires exclusion of infectious, neoplastic, and other autoimmune diseases. Laboratory tests are non-specific and reflect heightened immunological activity. Treatment comprises non-steroidal anti-inflammatory drugs, corticosteroids, immunosuppressive drugs (methotrexate, leflunomide, gold, azathioprine, cyclosporin A, cyclophosphamide), and intravenous gammaglobulin. The recent successful application of biological agents (anti-tumour necrosis factor, anti-interleukin (IL)1, anti-IL6), often in combination with traditional immunosuppressive drugs, has been very promising. CONCLUSIONS: AOSD often poses a diagnostic and therapeutic challenge and clinical guidelines are lacking. The emergence of validated diagnostic criteria, discovery of better serological markers, and the application of new biological agents may all provide the clinician with significant tools for the diagnosis and management of this complex systemic disorder. PMID- 16219708 TI - Preferential type 1 chemokine receptors and cytokine production of CD28- T cells in ankylosing spondylitis. AB - OBJECTIVE: To examine serum levels of type 1 and type 2 chemokines and lymphocytic expression of chemokine receptors, and to compare the results with lymphocytic cytokine production in patients with ankylosing spondylitis (AS). METHODS: Twelve patients with AS (mean (SD) age 44.9 (14.7) years) and 27 healthy controls (46.4 (12.8) years) were enrolled into the study. The expression of chemokine receptors (CCR-5, CXCR-3, CCR-4) and cytokines (interferon gamma (IFNgamma), interleukin (IL)2, IL4, IL10, tumour necrosis factor alpha (TNFalpha)) on CD28(+) and CD28(-) T cell subtypes was analysed by a three colour FACS technique of peripheral blood samples. Serum ELISAs were performed to detect the CCR-5 ligands CCL-5, CCL-3; the CXCR-3 ligands CXCL-10, CXCL-9; and the CCR-4 ligand, CCL-17 before and after administration of the TNFalpha blocking agent infliximab. RESULTS: CD4(+)CD28(-) T cells had higher ratios of CXCR-3 to CCR-4 than CD4(+)CD28(+) T cells. Both, CD4(+) and CD8(+)CD28(-) T cells of patients with AS produced more IFNgamma, TNFalpha, and IL10 than their CD28(+) counterparts (p<0.05), and lacked the production of IL2 and IL4. Serum levels of CXCL-9 were increased in patients with AS to 59.2 pg/ml (34.1-730.5) compared with 32.5 pg/ml (20.0-79.5) in healthy controls (p = 0.016). The levels of both type 1 (CCL-5, CXCL-9) and type 2 chemokines (CCL-17) decreased under blockade of TNFalpha (p<0.05). CONCLUSIONS: The profile of chemokine receptor expression and cytokine production by CD28(-) T cells suggests a type 1 immune reaction in AS, although IL10 is frequently produced by CD28(-) T cells. Treatment with TNFalpha blocking antibodies decreased both types of chemokines in patients' sera. PMID- 16219709 TI - Twenty-eight-joint counts invalidate the DAS28 remission definition owing to the omission of the lower extremity joints: a comparison with the original DAS remission. AB - OBJECTIVE: To compare 28 joint disease activity score (DAS28) remission with comprehensive joint count DAS remission in rheumatoid arthritis. METHODS: 620 actually measured paired observations of DAS28 and DAS were analysed in 155 patients. Discordant observations (either DAS or DAS28 below remission cut off level: 1.6 for DAS and 2.6 for DAS28) and concordant observations (both DAS and DAS28 below their remission cut off level) were analysed separately. RESULTS: 91 of 620 paired DAS observations (15%) were discordant; 87 (in 53 patients) comprised observations in which the DAS28 remission criterion, but not the DAS remission criterion, was met. The reverse was found in only four observations, which were therefore omitted. With the original DAS as standard, DAS28 sensitivity was 95% and specificity 84%. Probability plots showed a swollen joint count >0 in 75% of discordant pairs v 48% of concordant pairs. The same was found for total joint count (TJC >0 in 90% v 40%; median TJC, 0 v 6) and patient global assessment, but not for ESR. Individual joint analysis showed that 51% of discordant v 18% of concordant observations (p<0.0005) had involvement of lower extremity joints that are not included in the DAS28. CONCLUSIONS: DAS remission is more conservative than DAS28 remission. Activity (tenderness and swelling) in joints not included in the reduced joint counts (ankles, feet) mainly account for the discrepancy between the two assessments. DAS28 remission at a cut off level of 2.6 has insufficient construct validity and should be used with caution in clinical practice and clinical trials. PMID- 16219710 TI - The relation between progressive osteoarthritis of the knee and long term progression of osteoarthritis of the hand, hip, and lumbar spine. AB - BACKGROUND: The association between progression of knee osteoarthritis and progression of osteoarthritis at sites distant from the knee is unclear because of a lack of multisite longitudinal progression data. OBJECTIVE: To examine the association between radiological progression of knee osteoarthritis and osteoarthritis of the hands, hips, and lumbar spine in a population based cohort. METHODS: 914 women had knee x rays taken 10 years apart, which were read for the presence of osteophytes and joint space narrowing (JSN). Progression status was available for hand, hip, and lumbar spine x rays over the same 8 to 10 year period. The association between progression of knee osteoarthritis and osteoarthritis at other sites was analysed using odds ratios (OR) and 95% confidence intervals (CI) in logistic regression models. RESULTS: 89 of 133 women had progression of knee osteoarthritis based on osteophytes, and 51 of 148 based on JSN definition. Progression of JSN in the knee was predicted by progression in lumbar spine disc space narrowing (OR = 2.9 (95% CI 1.2 to 7.5)) and hip JSN (OR = 2.0 (1.0 to 4.2)). No consistent effects were seen for hand osteoarthritis. The associations remained after adjustment for age and body mass index. CONCLUSIONS: Progression of knee osteoarthritis is associated with progression of lumbar spine and hip osteoarthritis. This may have implications for trial methodology, the selection of patients for osteoarthritis research, and advice for patients on prognosis of osteoarthritis. PMID- 16219711 TI - "Joining the dots" for patients with systemic lupus erythematosus: personal perspectives of health care from a qualitative study. AB - OBJECTIVES: To examine the perceptions of patients with systemic lupus erythematosus (SLE) about their health care provision in the United Kingdom. METHODS: Semistructured interviews were conducted with 10 women aged 26 to 68 years who were diagnosed with SLE one to 12 years earlier. Interviews were audio recorded, transcribed verbatim, and analysed using interpretative phenomenological analysis to organise the themes of importance to participants. RESULTS: Four themes emerged: diagnostic difficulties; understanding; communication; and integrated health care. Before diagnosis there was concern to appear legitimately ill and to have a label for the condition. After diagnosis participants still encountered health care professionals who were poorly informed about SLE. Family, friends, and employers did not understand the fluctuating nature of SLE, which often led to isolation. Participants felt that even health care professionals who specialised in SLE could not fully understand the psychosocial impact of the condition, and therefore did not provide information to meet those needs. Participants did not know which of the many health care professionals they had contact with to approach about their concerns. Lack of communication at an interdisciplinary level left them feeling that nobody was "joining the dots" for their health care. CONCLUSIONS: Patients with SLE do not feel understood by health care providers or people close to them. Support from trained volunteers with SLE, as available at the open access lupus clinic in Dudley (West Midlands, UK), would ensure more adequate information from someone with personal experience. Such services may improve communication and help minimise SLE patients' isolation. PMID- 16219712 TI - Impaired vascular reactivity in African-American patients with type 2 diabetes mellitus and microalbuminuria or proteinuria despite angiotensin-converting enzyme inhibitor therapy. AB - BACKGROUND: Microalbuminuria, an early indicator of diabetic nephropathy that reflects other vascular abnormalities, usually improves or resolves with angiotensin-converting enzyme inhibitor (ACEI) therapy. Persistent microalbuminuria despite ACEI therapy may be associated with poor prognosis for cardiovascular disease and mortality. African-Americans are reported to respond less well to ACEI and are at increased risk of disease progression. METHODS AND RESULTS: We compared flow-mediated dilatation (FMD) and nitroglycerine-dependent dilatation (NDD) in African-American diabetic subjects with persistent microalbuminuria (n = 35) despite ACEI therapy and those in whom microalbuminuria had resolved (n = 15). The two groups were not statistically different in terms of blood pressure, age, sex, lipids, and hemoglobin A1c. FMD was reduced in the microalbuminuria group, compared with subjects without microalbuminuria (4.2 vs. 11.4%; P < 0.0001). Similarly, NDD was reduced in the microalbuminuria group, compared with subjects without microalbuminuria (10.8 vs.16.6%; P = 0.011). The FMD in African-American patients with persistent microalbuminuria was also significantly lower than in clinically similar Caucasian patients whose microalbuminuria had persisted despite ACEI therapy (4.2 vs. 7.5%; P = 0.03). On multiple regression analysis, persistent microalbuminuria is the only predictor of abnormal endothelial function in these patients. CONCLUSIONS: Our study clearly demonstrates that African-American type 2 diabetic subjects with persistent microalbuminuria have severely impaired FMD and NDD, compared with matched patients who had microalbuminuria that was eliminated by ACEI. This may explain the poor prognosis for cardiovascular disease in patients who have persistent microalbuminuria. Alternative strategies for reducing microalbuminuria in high-risk patients who do not respond adequately to ACEI therapy such as African-Americans are needed. PMID- 16219713 TI - Acute regulation of circulating parathyroid hormone (PTH) molecular forms by calcium: utility of PTH fragments/PTH(1-84) ratios derived from three generations of PTH assays. AB - CONTEXT: The quantitative evaluation of circulating PTH peaks revealed by PTH assays after HPLC separation constitutes the best way to study the behavior of PTH molecular forms, but it is also impractical. OBJECTIVE: The objective of the study was to investigate the regulation of circulating PTH molecular forms by calcium through the use of PTH fragments/PTH (1-84) ratios derived from PTH assays with different specificities before and after HPLC separation of circulating PTH. DESIGN: CaCl2 and Na citrate were infused in eight volunteers. PTH was measured in serum and HPLC fractions at different calcium concentrations in PTH assays reacting with regions 1-2 (CA), 12-18 (T), and 65-69 (C) of the PTH structure. RESULTS: From hypo- to hypercalcemia, the C/CA ratio had the highest range (1.92 to 9.75; P < 0.001), and the C/T ratio had a higher range (1.69 to 6.11; P < 0.01) than the T/CA ratio (1.15 to 1.86). Human (h) PTH (1-84) represented 32.7 and 4.3% of circulating PTH in hypo- and hypercalcemic HPLC profiles, respectively. These numbers were 5 and 0.9% for amino-terminal (N)-PTH, an amino-terminal form of PTH distinct from hPTH (1-84), 7.3 and 6.8% for non-(1 84) PTH or large C-PTH fragments with a partially preserved N structure, and 54.9 and 88.1% for C-PTH fragments missing a N structure. The HPLC C-PTH fragments to hPTH (1-84) ratio had the most extensive range (1.67 to 20.58). Despite their quantitative differences, all ratios identified identical behavior of PTH fragments relative to PTH (1-84). CONCLUSIONS: PTH assay ratios are an adequate tool to investigate the modulation of PTH molecular forms, even if all PTH assays show some undesirable cross-reactivity with certain circulating forms of PTH. PMID- 16219714 TI - Heritability of polycystic ovary syndrome in a Dutch twin-family study. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders among women of reproductive age. There is evidence for a genetic component in PCOS based on familial clustering of cases. OBJECTIVE: In the present study, the heritability of PCOS was estimated. DESIGN/PARTICIPANTS: Data from 1332 monozygotic twins (genetically identical) and 1873 dizygotic twins/singleton sisters of twins (who share on average 50% of their segregating genes) registered with The Netherlands Twin Register were used. PCOS was defined as less than nine menstrual cycles and acne or hirsutism in agreement with the 2003 Rotterdam consensus. RESULTS: Results point to a strong contribution of familial factors to PCOS. The resemblance in monozygotic twin sisters (tetrachoric correlation 0.71) for PCOS was about twice as large as in dizygotic twin and other sisters (tetrachoric correlation 0.38). Univariate analyses point to strong contributions of genetic factors to the variance in PCOS. Next, a trivariate genetic analysis of oligomenorrhea, acne, and hirsutism was carried out. This analysis confirmed that the familial component in PCOS is due to genetic factors. CONCLUSIONS: This study demonstrated a large influence of genetic factors to the pathogenesis of PCOS, justifying the search for susceptibility genes. PMID- 16219715 TI - PAX8-PPARgamma rearrangement is frequently detected in the follicular variant of papillary thyroid carcinoma. AB - CONTEXT: The clinicopathological characteristics and the molecular features of the follicular variant of papillary thyroid carcinoma (FVPTC) remain controversial. OBJECTIVE/DESIGN/PATIENTS: In an attempt to clarify such controversies and to find whether or not FVPTC cases share the molecular features of follicular tumors, we searched for the presence of PAX8-PPARgamma rearrangements, RAS mutations, and RAP-1, RAF-1, and BRAF mutations in a series of 40 FVPTCs as well as in 27 follicular thyroid carcinomas (FTCs) and 12 follicular thyroid adenomas (FTAs). Fluorescence in situ hybridization and RT-PCR were used to detect the PAX8-PPARgamma rearrangement and PCR, single strand confirmational polymorphism, and sequencing for searching the mutations. RESULTS: The frequency of PAX8-PPARgamma rearrangement was similar in FVPTCs (37.5%), FTCs (45.5%), and FTAs (33.3%). The same holds true regarding the frequency and type of RAS mutations: FVPTC, 25.0%; FTC, 22.2%; and FTA, 33.3%. BRAF mutations were only detected in FVPTC (10%); the BRAF mutations in these cases (K601E and G474R) are different from the typical BRAF(V600E) mutation of conventional PTCs. No mutations were detected in RAP-1 and RAF-1. In FVPTCs, the PAX8-PPARgamma rearrangement was significantly associated with multifocality and vascular invasion, whereas the RAS mutations were significantly associated with the large tumor size. There were three cases of FVPTC, three FTCs and one FTA, harboring both PAX8-PPARgamma rearrangement and RAS mutations; patients with such tumors were usually very young. CONCLUSIONS: We conclude that a subset of FVPTC shares some of the molecular features of follicular tumors. Further studies are necessary to clarify the putative clinical significance (e.g. association to blood-born metastases) of PAX8-PPARgamma rearrangement, RAS mutations, and BRAF(K601E) in FVPTCs. PMID- 16219716 TI - Comparative evaluation of recombinant human thyrotropin-stimulated thyroglobulin levels, 131I whole-body scintigraphy, and neck ultrasonography in the follow-up of patients with papillary thyroid microcarcinoma who have not undergone radioiodine therapy. AB - CONTEXT: Although the prognosis of papillary thyroid microcarcinoma (PTMC) is usually excellent, the optimal follow-up strategy has never been investigated. OBJECTIVE: The objective of the study was to investigate the role of neck ultrasonography (US), whole-body scintigraphy (WBS), and serum thyroglobulin levels (Tg) after recombinant human (rh) TSH in the follow-up of very low-risk PTMC patients. DESIGN: The study was a 5-yr observational study based on a 6- to 12-month follow-up after near total thyroidectomy. SETTING: The study population consisted of ambulatory patients. PATIENTS: Eighty consecutive patients diagnosed with PTMC, who had not undergone postoperative radioiodine treatment because of unifocal tumor without lymph node metastases and who did not have anti-Tg antibodies, were included. MAIN OUTCOME MEASURES: WBS and Tg after both rhTSH and neck US were measured. RESULTS: rhTSH-Tg was 1 ng/ml or less in 45 (Tg-) and more than 1 in 35 (Tg+) patients. WBS showed no pathological uptake in any patient. US identified node metastases in two Tg (+) and one Tg (-) patients. rhTSH-Tg levels positively correlated with thyroid bed iodine uptake (r = 0.40, P < 0.0001). To date (32 +/- 13 months after surgery), all node-negative patients have undetectable Tg levels on LT(4) treatment and negative US. CONCLUSIONS: For the initial follow-up of PTMC patients without risk factors and anti-Tg antibodies and who did not undergo radioiodine treatment: 1) WBS is useless; 2) US is highly sensitive in detecting node metastases; and 3) detectable rhTSH-Tg levels mainly depend on small normal tissue remnants. In this subgroup of PTMC patients, neck US might be regarded as a primary tool for the initial follow-up. PMID- 16219717 TI - Adiponectin is inversely associated with renal function in type 1 diabetic patients. AB - OBJECTIVE: Adipose tissue is a source of several adipocytokines that may contribute to vascular complications. We examined the relation of adiponectin with several cardiovascular risk factors and with micro- and macrovascular outcomes in type 1 diabetic patients. DESIGN: Cross-sectional data on 543 type 1 diabetic patients from the EURODIAB Prospective Complications Study were analyzed. We determined adiponectin, TNF-alpha, IL-6, C-reactive protein, soluble vascular cell adhesion molecule (sVCAM-1), and sE-selectin by ELISA. RESULTS: We found that adiponectin was negatively correlated with body mass index, waist to hip ratio, insulin, and fasting triglyceride, and positively with high-density lipoprotein, low-density lipoprotein, and total cholesterol, TNF-alpha, and sVCAM 1, but was not related to C-reactive protein, IL-6, and sE-selectin. Surprisingly, significantly raised concentrations of adiponectin were found with albuminuria, retinopathy, and cardiovascular diseases (for all, P < 0.0001). Adiponectin levels were inversely associated with glomerular filtration rate (GFR) (P < 0.0001). Multivariate regression models showed that the associations of adiponectin with albuminuria and GFR were independent of established risk factors. The association between adiponectin and albuminuria was attenuated by GFR, whereas the association of adiponectin with retinopathy and cardiovascular disease disappeared after adjustments for established risk factors. The association of adiponectin with sVCAM-1 was independent of established risk factors. CONCLUSION: We conclude that in type 1 diabetic patients, adiponectin is associated with impaired renal function. Adiponectin may be enhanced in type 1 diabetic patients as a physiological counterregulatory response to mitigate endothelial damage and vascular damage. PMID- 16219718 TI - An assessment of petrosal sinus sampling for localization of pituitary microadenomas in children with Cushing disease. AB - CONTEXT: Pituitary adenomas in Cushing disease (CD) are usually small and difficult to visualize. Bilateral inferior petrosal venous sampling (BIPSS) before and after ovine CRH stimulation is reserved for patients who have ACTH dependent Cushing syndrome and negative magnetic resonance imaging (MRI) or positive MRI but inconsistent biochemical data. OBJECTIVE: The objective of the study was to evaluate the usefulness of BIPSS as a tool for localization of a pituitary adenoma in children with CD. DESIGN: The study was a retrospective review of the records of 141 children who were admitted for evaluation of CD from 1982 to 2004. SETTING: The study was conducted at a tertiary care center. INTERVENTIONS AND OUTCOME MEASURES: Lateralization of ACTH secretion during BIPSS was compared with MRI and surgical findings for the localization of a microadenoma. RESULTS: A total of 94 patients, 49 males and 45 females with an age range of 5.3 to 18.7 yr (13 +/- 3.2 yr), underwent BIPSS. Localization of a microadenoma by BIPSS agreed with surgical location in only 58% of the cases (95% confidence interval, 43-66). The combined use of information from the MRI and inferior petrosal venous sampling did not predict the location of the tumor more frequently than MRI alone (P > 0.1), which in this study localized a lesion in 39% of the patients (95% confidence interval, 28-50). The procedure was completed successfully in all patients, and no serious complications were recorded. CONCLUSIONS: Although BIPSS was safe and well tolerated in an experienced center, lateralization of the ACTH gradient during BIPSS was a poor predictor of the site of the adenoma in children with CD. PMID- 16219719 TI - Relationships between sleep disordered breathing and glucose metabolism in polycystic ovary syndrome. AB - CONTEXT: Women with polycystic ovary syndrome (PCOS) are insulin resistant and are at increased risk for sleep apnea, which, in turn, may contribute to insulin resistance. OBJECTIVE: The objective of this study was to determine relationships between risk and severity of obstructive sleep apnea (OSA) and glucose metabolism in PCOS. DESIGN AND SETTING: This study included two cohorts of women with PCOS in a tertiary care hospital. PATIENTS AND MAIN OUTCOME MEASURES: Cohort 1 included 40 nondiabetics who completed the Epworth Sleepiness Scale, the Pittsburgh Sleep Quality, and the Berlin Questionnaire to assess risk of OSA; 32 of the 40 women had an oral glucose tolerance test. Cohort 2 included eight women who had a sleep study, glycosylated hemoglobin level, and an oral glucose tolerance test. RESULTS: In cohort 1, 62.5% of the women had poor sleep quality by Pittsburgh Sleep Quality Index, and 18 (45%) had chronic daytime sleepiness by Epworth Sleepiness Scale. Thirty of the 40 women had a high risk of OSA by Berlin Questionnaire. Women with high OSA risk had higher fasting insulin levels and homeostasis model assessment index compared with those with low OSA risk (168.2 +/- 17.3 vs. 97.2 +/- 6.4 pmol/liter, P = 0.011; 6.3 +/- 0.7 vs. 3.6 +/- 0.3 mg/dl x microU/ml, P = 0.014, respectively). Among women with normal glucose tolerance, insulin levels were significantly higher in those at high vs. low OSA risk, independently of body mass index. Women in cohort 2 had rapid eye movement (REM)-predominant OSA with lower sleep efficiency, longer sleep latency, and less REM sleep than controls. Glycosylated hemoglobin levels and the area under the glucose curve positively correlated with the apnea-hypopnea index (rP = 0.82, P = 0.013; rP = 0.96, P = 0.0008, respectively) and the number of oxygen desaturations in REM sleep (rP = 0.97, P = 0.0009; rP = 0.97, P = 0.005, respectively). CONCLUSION: PCOS is associated with poor sleep quality, daytime sleepiness, and increased risk for OSA. Insulin levels and measures of glucose tolerance in PCOS are strongly correlated with the risk and severity of OSA. PMID- 16219720 TI - Thyrotropin suppression by metformin. AB - CONTEXT: Drug-drug interactions are common but often are discovered only long after initial drug release. Metformin has been available in the United States for 9 yr and elsewhere for many years, but as of yet there are no reports that the drug modifies thyroid hormone economy. OBJECTIVE: The objective of the study was to describe the clinical and biochemical findings of four patients with chronic hypothyroidism, previously euthyroid on fixed doses of L-T4 for several years, in whom the metformin was initiated. DESIGN: This was a retrospective review. SETTING: The study was conducted at a tertiary care military hospital providing care to active-duty soldiers, sailors, and marines, retirees of the armed forces, and their eligible dependents. PARTICIPANTS: Four patients with chronic hypothyroidism who were placed on metformin participated in the study. INTERVENTION, MAIN OUTCOME MEASURE: Serum TSH, free T4, and free T3 levels were measured during metformin treatment. RESULTS: Initiation of treatment with metformin (three for diabetes mellitus and one for nonalcoholic steatohepatitis) caused suppression of TSH to subnormal levels without clinical symptoms of hyperthyroidism in any patients. There was no change in free T4 or free T3 in patient 1. CONCLUSIONS: No other potential causes of TSH suppression, including medication changes or interference in the TSH assay, could be identified. The mechanism of the fall in serum TSH in these four patients is unclear at this time. Should these findings be confirmed in larger prospective studies, metformin's ability to suppress TSH without causing clinical or chemical hyperthyroidism might render this drug a useful adjunct to the treatment of patients with thyroid cancer. PMID- 16219721 TI - Keratoconus managed with intacs: one-year results. AB - OBJECTIVES: To describe the visual outcome of keratoconus managed with Intacs implantation (Addition Technology Inc, Fremont, Calif) and to define criteria that predict good outcome. METHODS: This retrospective, nonrandomized, comparative, consecutive case series studied 58 eyes of 43 patients with keratoconus managed by Intacs implantation. The outcome measures were analyzed pre-Intacs and 1 year post-Intacs. Preoperative parameters were correlated with outcome. MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA), best spectacle corrected visual acuity (BSCVA), manifest refraction, videokeratography, and patient questionnaires. RESULTS: Intacs were implanted in all eyes with no intraoperative complications. Six eyes underwent additional Intacs surgery. Post Intacs, the mean +/- SD UCVA improved from less than 20/200 +/- 0.1 line to 20/50(-3) +/- 3.1 lines, the mean +/- SD BSCVA was unchanged at 20/32 +/- 2.0 lines, the mean +/- SD spherical equivalent improved from -3.88 +/- 1.64 to -1.04 +/- 1.51 diopters (D), and the mean +/- SD astigmatism improved from 3.34 +/- 2.23 to 1.97 +/- 1.51 D. Twenty-five eyes had a good outcome (UCVA> or =20/40). Multiple regression selected BSCVA, astigmatism, and spherical myopia as the preoperative predictors of outcome. CONCLUSIONS: Intacs improve myopia and regular astigmatism in keratoconus. Milder keratoconus (BSCVA>20/32(-2) and astigmatism<3.50 D) and significant spherical myopia (>-1.75) predict better outcome. PMID- 16219722 TI - Modified osteo-odonto-keratoprosthesis for treatment of corneal blindness: long term anatomical and functional outcomes in 181 cases. AB - OBJECTIVE: To evaluate long-term anatomical and functional outcomes of a modified osteo-odonto-keratoprosthesis (OOKP) technique for treatment of corneal blindness from various etiologies. METHODS: Two-hundred three patients (224 eyes) underwent modified OOKP surgery between 1973 and 1999. Of the original cohort, 181 patients (98 men and 83 women; mean [SD], age 54.3 [15] years) in whom a standardized 2 step surgical procedure was performed were included in the study. Preoperative diagnoses were dry eye (n = 70) due to ocular pemphigoid (n = 39), Sjogren syndrome (n = 11), trachoma (n = 8), Lyell syndrome (n = 6), Stevens-Johnson syndrome (n = 4), and graft-vs-host disease (n = 1) and congenital lid coloboma (n = 1), severe corneal burns (n = 68), bullous keratopathy (n = 13), keratitis sequelae (n = 15), and bullous keratopathy secondary to antiglaucoma surgery (n = 15). Several innovations were made to the original Strampelli technique. Median follow-up duration was 12 years (range, 1-25 years). RESULTS: Anatomical complications leading to OOKP loss were found in 11 (6.07%) of 181 patients. Survival analysis estimated that 18 years after surgery, the probability of retaining an intact OOKP was 85% (95% confidence interval, 79.3%-90.7%). Pooling patient groups, mean (SD) best postoperative visual acuity was 0.76 (0.34). Mean (SD) final acuity at the end of follow-up declined slightly (0.69 [0.39]) but significantly (P<.01). In individual diagnostic groups, mean acuity decline reached statistical significance (P<.05) only in the pemphigoid (1 line), trachoma (1 line), and bullous keratopathy secondary to antiglaucoma surgery (2 lines) groups. Survival analysis estimated that 18 years after surgery, the probability of retaining best postoperative visual acuity (within 2 lines) was mean (SD) 55.5% (12.9%). CONCLUSION: Modified OOKP surgery for corneal blindness of different etiologies may provide, in the long-term, anatomically stable corneal prosthesis as well as an effective, rehabilitating recovery in visual acuity. PMID- 16219723 TI - Repeatability and reproducibility of fast macular thickness mapping with stratus optical coherence tomography. AB - OBJECTIVE: To determine the repeatability and reproducibility of retinal thickness measurements with the fast macular thickness mapping protocol of Stratus optical coherence tomography. METHODS: Ten eyes of 10 healthy subjects and 15 eyes of 15 diabetic patients with clinically significant macular edema (CSME) underwent 2 scanning sessions before and after pupil dilation during the same visit by 2 experienced examiners. Healthy subjects also received a third scanning session during a second visit. Repeatability and reproducibility for the foveolar center and for each of 9 Early Treatment of Diabetic Retinopathy Study (ETDRS)-like regions were calculated by their repeatability and reproducibility coefficients and intraclass correlation coefficients. RESULTS: The coefficients of repeatability were less than 8% in healthy subjects and less than 9% in patients with diabetes and CSME. The reproducibility coefficients were less than 10% and 11% in healthy subjects and diabetic patients with CSME, respectively. There was no significant difference between scans acquired by different observers or during different visits. The intraclass correlation coefficients were always greater than 0.80 and 0.98 in healthy subjects and diabetic patients with CSME, respectively. Average +/- SD thickness was found to be 223 +/- 14 and 404 +/- 108 mum for the central ETDRS-like region in healthy subjects and diabetic patients with CSME, respectively. CONCLUSION: With the fast macular thickness mapping protocol of Stratus ocular coherence tomography, our results indicate that retinal thickness measurements in dilated and undilated eyes of healthy subjects and diabetic patients with CSME are repeatable and reproducible. PMID- 16219724 TI - Predictive factors for visual acuity after intravitreal triamcinolone treatment for diabetic macular edema. AB - OBJECTIVE: To evaluate which factors influence maximum gain in best-corrected visual acuity after intravitreal injection of triamcinolone acetonide as treatment for diffuse diabetic macular edema. METHODS: This prospective clinical interventional study included 53 eyes with diffuse diabetic macular edema receiving an intravitreal injection of about 20 mg of triamcinolone. The mean +/- SD follow-up was 10.2 +/- 7.6 months. RESULTS: In a multiple linear regression analysis, maximum gain in best-corrected visual acuity after the intravitreal injection of triamcinolone was significantly (P < .001) and negatively correlated with an increased degree of macular ischemia and a higher preoperative visual acuity. Improvement in best-corrected visual acuity was significantly and positively correlated with increased degree of macular edema (P = .001). Change in best-corrected visual acuity after the intravitreal triamcinolone injection was statistically independent (P > .15) of age, sex, pseudophakia, and macula grid laser treatment before inclusion into the study. The results were comparable for gain in visual acuity at 6 months after the injection. CONCLUSION: Pronounced macular edema may have a positive impact, and marked macular ischemia and a high preoperative best-corrected visual acuity may have a negative impact, on an increase in best-corrected visual acuity after intravitreal triamcinolone injection in patients with diabetic macular edema. PMID- 16219725 TI - Complications after photodynamic therapy. AB - OBJECTIVE: To evaluate the incidence of complications of photodynamic therapy (PDT) with verteporfin in subfoveal choroidal neovascularizations secondary to age-related macular degeneration and pathologic myopia. METHODS: In this retrospective interventional case series, the occurrence of complications after PDT in a clinical setting was analyzed. Consecutive medical records of patients with age-related macular degeneration and pathologic myopia treated with PDT were reviewed for complications. Complications included treatment-related systemic adverse events, injection site effects, and ocular adverse events. RESULTS: We included 273 patients (198 with age-related macular degeneration and 75 with pathologic myopia) in the study. A total of 485 photodynamic treatment sessions were performed. Infusion-related back or chest pain was reported by 6 patients (2.2%; 95% confidence interval [CI], 0.8%-4.7%). Injection site effects, extravasation, and photosensitivity reactions were not observed. Dyspnea and flushing during infusion were observed in 2 patients (0.7%; 95% CI, 0.09%-2.6%). Body pain, shortness of breath, and elevated blood pressure were noted in 13 patients (4.8%; 95% CI, 2.6%-8.0%). General pruritus was described by 6 patients (2.2%; 95% CI, 0.8%-4.7%), starting 4 hours after the infusion of verteporfin, and resolved within 72 hours after PDT. A total of 8 patients (2.9%; 95% CI, 1.3% 5.7%) reported an acute severe visual acuity decrease of at least 4 Early Treatment Diabetic Retinopathy Study lines occurring within 7 days of treatment. CONCLUSIONS: Complications associated with PDT are uncommon, but there were limitations of retrospective studies for identifying safety problems. Complications like acute severe visual events may occur in about 3% of patients. We believe that this risk is outweighed by the benefits of PDT on visual function in most patients. PMID- 16219726 TI - Validation of a predictive model to estimate the risk of conversion from ocular hypertension to glaucoma. AB - OBJECTIVES: To develop and validate a predictive model to estimate the risk of conversion from ocular hypertension to glaucoma. METHODS: Predictive models for the 5-year risk of conversion to glaucoma were derived from the results of the Ocular Hypertension Treatment Study (OHTS). The performance of these models was assessed in an independent population of 126 subjects with ocular hypertension from a longitudinal study (Diagnostic Innovations in Glaucoma Study [DIGS]). The performance of the OHTS-derived models was assessed in the DIGS cohort according to equality of regression coefficients, discrimination (c-index), and calibration. RESULTS: Thirty-one patients (25%) developed glaucoma during follow up. Hazard ratios for DIGS- and OHTS-derived predictive models were similar for age, intraocular pressure, central corneal thickness, vertical cup-disc ratio, and pattern standard deviation but were significantly different for the presence of diabetes mellitus. When applied to the DIGS population, the OHTS-derived predictive models had reasonably good discrimination (c-indexes of 0.68 [full model] and 0.73 [reduced model]) and calibration. CONCLUSIONS: The OHTS-derived predictive models performed well in assessing the risk of glaucoma development in an independent population of untreated subjects with ocular hypertension. A risk scoring system was developed that allows calculation of the 5-year risk of glaucoma development for an individual patient. PMID- 16219727 TI - Reduced melanoma-related mortality in uveal melanoma by preenucleation radiotherapy. AB - BACKGROUND: Radiotherapy of an eye before enucleation, so called preenucleation radiotherapy (PER), of patients with uveal melanoma was initiated to reduce enucleation-induced systemic metastasis. Earlier studies with a short follow-up period have not demonstrated a significant effect on survival. OBJECTIVE: To study the effect of PER on melanoma-related mortality after more than 9 years of follow-up. DESIGN: In a prospective study, 167 patients with uveal melanoma were treated between 1978 and 1992 by irradiation with 800 rad (8 Gy) given in 2 fractions 2 days before enucleation. A group of 108 patients with uveal melanoma treated between 1971 and 1992 by enucleation only in the same hospital served as a historical control group. Patients were followed up until December 2002 or death. RESULTS: Melanoma-related death occurred in 32.3% of the PER-treated group and in 40.7% of the enucleation only group. Mean follow-up was 9.25 years. After 48 months of follow-up, a significant difference in survival became evident in favor of the PER group. The estimated 15-year survival rates for patients with melanoma in the PER group and enucleation only group were 63.7% and 51.0%, respectively. For patients dying of all causes, these percentages were 47.5% and 25.2%, respectively. In both groups, women had a better prognostic outcome than men. CONCLUSION: This study suggests that PER improves long-term survival in patients with uveal melanoma. PMID- 16219728 TI - Peripapillary staphyloma: clinical features and visual outcome in 19 cases. AB - OBJECTIVE: To investigate the clinical features and visual outcome of patients with peripapillary staphyloma. METHODS: The medical records of patients diagnosed as having peripapillary staphyloma at Seoul National University Children's Hospital, Seoul, Korea, between January 1, 1991, and December 31, 2003, were reviewed. Visual acuity and associated ocular and systemic abnormalities were recorded. RESULTS: Nineteen patients (21 eyes) with peripapillary staphyloma were included in the study. The mean age at initial examination was 21 months, and 42% (8/19) of the patients were girls. Two patients (11%) had bilateral peripapillary staphylomas. Only 2 eyes had visual acuity better than 20/200 at the last examination, and 7 eyes had severe myopia of more than -6 diopters. After occlusion therapy, 1 patient achieved a visual improvement from light perception to 20/30. Congenital cataract and persistent pupillary membrane were present in 2 eyes each at initial examination. Complications observed during the follow-up period included acquired total cataract in 2 eyes and retinal detachment in 3 eyes. Scleral encircling buckling was performed in 1 eye with retinal detachment, and subsequently the retina was reattached. Only 2 patients had accompanying systemic abnormalities. CONCLUSIONS: Rarely, patients with peripapillary staphyloma can achieve significant visual improvement by occlusion therapy, although visual outcome is generally poor. Because associated ocular disease and refractive errors are not infrequent, complete ophthalmic examinations and regular follow-up are necessary. PMID- 16219729 TI - Selective death of lens epithelial cells using demineralized water and Triton X 100 with PerfectCapsule sealed capsule irrigation: a histological study in rabbit eyes. AB - BACKGROUND: Sealed irrigation of the capsular bag after removal of the cataractous lens isolates the interior of the capsular bag from the anterior segment, permitting isolated targeting of lens epithelial cells (LECs) in vivo using pharmacologic agents while minimizing the risk of damage to other intraocular structures. OBJECTIVES: To assess the ability to deliver a nonspecific, extremely toxic agent (Triton X-100) directly to the LECs after crystalline lens removal, and to assess the eyes histologically for evidence of collateral damage. METHODS: Twelve eyes from 6 New Zealand White rabbits were divided into 3 groups of 4 eyes that underwent phacoaspiration of the crystalline lens. Group 1 was a control. In group 2, the anterior segment was irrigated with Triton X-100 and demineralized water for injection for 5 minutes. In group 3, the lens capsule was isolated from the anterior segment using sealed capsule irrigation (SCI) with PerfectCapsule (Milvella Pty Ltd, Sydney, Australia). After surgery, the rabbits were humanely euthanized. The enucleated eyes were immediately fixed in 10% neutral buffered formalin, histological analysis was performed to assess the corneal endothelium, iris, and retina, and the capsular bag and residual equatorial LECs were assessed. RESULTS: The capsular bag was sealed and inflated under SCI in all treated eyes in group 3. Histological evaluation revealed no evidence of collateral damage in group 1 and group 3. Significant histological damage to the cornea, iris, and peripheral retina was noted in group 2. Histological evaluation of each capsular bag suggests presence of LECs in group 1 and group 2. In the presence of SCI, Triton X-100 caused almost complete destruction of LECs in the capsular bag. CONCLUSIONS: Results suggest that SCI allows selective delivery of toxic agents directly into the capsular bag, preventing collateral damage to surrounding intraocular structures. The PerfectCapsule device kept the capsular bag well inflated intraoperatively, which may allow the isolated, safe delivery of pharmacologic agents into the capsular bag during cataract surgery. Clinical Relevance Postoperative proliferation of LECs in the capsular bag remains the most frequent complication of cataract surgery. Unprotected intraocular infusion of cytotoxic drugs, antimetabolites, or hypo-osmotic agents during cataract surgery has the potential risk of causing toxic effects in corneal endothelium and adjacent intraocular tissues. Selective delivery of pharmacologic/hypo-osmotic agents into the capsular bag requires positive pressure inflation of the bag and is now possible using an SCI device. This device allows the surgeon to reseal the capsular bag intraoperatively, achieve positive pressure inflation of the capsular bag, and direct selective targeting of LECs. PMID- 16219730 TI - Retinal progenitor cell xenografts to the pig retina: morphologic integration and cytochemical differentiation. AB - OBJECTIVE: To investigate the survival, integration, and differentiation of mouse retinal progenitor cells after transplantation to the subretinal space of adult pigs. METHODS: Green fluorescent protein-positive (GFP+) murine retinal progenitor cells were transplanted subretinally as single cells, spheres, or biodegradable polymer-progenitor composites into 24 nonimmunosuppressed adult pigs. Of these, 14 pigs received laser lesions (n = 11) or outer retinal scraping injury (n = 3). Recipients were killed at 30 minutes to 5 weeks after grafting. RESULTS: The GFP+ murine retinal progenitor cells survived well for up to 14 days after transplantation to the pig retina. After 5 weeks, fewer GFP+ cells were found. In the pigs that received laser treatment before grafting of cell suspension, GFP+ cells integrated into the retinal pigment epithelium and all layers of the retina. The GFP+ cells exhibited morphologic evidence of differentiation into mature retinal neurons, although evaluation of marker expression found only nestin and glial fibrillary acidic protein colocalization. In noninjured pigs, cells mainly integrated into the retinal pigment epithelium. In pigs that received composites, cells appeared to mature and extended processes through pores in the polymer matrix. CONCLUSIONS: Retinal progenitor cell xenografts survive for a sufficiently long period to integrate into areas of injury and exhibit morphologic differentiation. By 5 weeks, survival diminishes. Biodegradable polymers may be useful for transplanting retinal progenitor cells in a structurally organized manner. Clinical Relevance Central nervous system (CNS) diseases may cause long-term disabilities. Substantial tissue destruction can be sustained by the complex structures of the brain, spinal cord, or retina without loss of life, yet the lack of effective CNS regeneration frequently results in disruption of activities of daily living and marked degradation in quality of life. It has become clear that an enormous potential for repair is present within the mammalian CNS. The challenge is to harness this potential to treat disease. Transplantation of neuronal tissue to the CNS represents a promising, albeit challenging, approach to the replacement of neurons lost owing to injury or disease. PMID- 16219731 TI - Cause-specific visual impairment and mortality: results from a population-based study of older people in the United Kingdom. AB - OBJECTIVE: To assess the association between mortality and cause-specific visual impairment in older people. METHODS: Visual acuity and causes of visual impairment were collected in 13 569 participants 75 years and older participating in a randomized trial of health screening. Participants were followed up for mortality for a median of 6.1 years. RESULTS: Compared with those with 6/6 (or 20/20 Snellen) or better visual acuity, the age- and sex-adjusted rate ratio for visually impaired people (binocular visual acuity <6/18 or <20/60 Snellen) was 1.60 (95% confidence interval, 1.47-1.74), which was markedly attenuated (rate ratio, 1.17; 95% confidence interval, 1.07-1.27) after adjustment for confounding factors. People whose visual impairment was due to cataract or age-related macular degeneration had excess risks of all-cause and cardiovascular mortality, which disappeared after adjustment. People with refractive error remained at small risk, despite adjustment, probably owing to residual confounding from factors associated with minimal use of eye services rather than underlying eye disease. There were no associations with cancer mortality. CONCLUSION: Associations reported for visual impairment and mortality or for specific causes of visual impairment reflect confounding by comorbidities, risk factors, and other factors related to susceptibility to death rather than an independent biological association of vision problems or specific eye diseases. PMID- 16219732 TI - Development of spherical equivalent refraction in prematurely born children during the first 10 years of life: a population-based study. AB - OBJECTIVE: To evaluate the development of refraction, expressed as spherical equivalents, in prematurely born children during the first 10 years of life. METHODS: Retinoscopy in cycloplegia was performed at 6 months, 2.5 years, and 10 years of age in 198 prematurely born children from a previous population-based study on the incidence of retinopathy of prematurity. Spherical equivalents were calculated. Myopia was defined as a spherical equivalent of less than 0 diopters (D), clinically significant myopia at 10 years of age as -1 D or less, and moderate or high myopia as less than -3D. Hypermetropia greater than +3 D was regarded as significant. RESULTS: There were no significant differences during the refractive development between the various subgroups of retinopathy of prematurity. Cryotreated eyes had a wider distribution of refractive errors. A multiple regression analysis revealed that the spherical equivalents at 2.5 years of age predicted clinically significant myopia (500 silent and 80 amino acid substitutions between these mtDNAs). Longevity was studied in reciprocal cross genotypes between pairs of these strains to test for cytoplasmic (mtDNA) factors affecting aging. The intrapopulation crosses between Zimbabwe strains show no difference in longevity between mtDNAs; the interpopulation crosses between Crete and the United States show subtle but significant differences in longevity; and the interspecific introgression lines showed very significant differences between mtDNAs. However, the genotypes carrying the D. simulans mtDNA were not consistently short-lived, as might be predicted from the disruption of nuclear-mitochondrial coadaptation. Rather, the interspecific mtDNA strains showed a wide range of variation that flanked the longevities seen between intraspecific mtDNAs, resulting in very significant nuclear x mtDNA epistatic interaction effects. These results suggest that even "defective" mtDNA haplotypes could extend longevity in different nuclear allelic backgrounds, which could account for the variable effects attributable to mtDNA haplogroups in human aging. PMID- 16219777 TI - Genetic analysis of Saccharomyces cerevisiae H2A serine 129 mutant suggests a functional relationship between H2A and the sister-chromatid cohesion partners Csm3-Tof1 for the repair of topoisomerase I-induced DNA damage. AB - Collision between a topoisomerase I-DNA intermediate and an advancing replication fork represents a unique form of replicative damage. We have shown previously that yeast H2A serine 129 is involved in the recovery from this type of damage. We now report that efficient repair also requires proteins involved in chromatid cohesion: Csm3; Tof1; Mrc1, and Dcc1. Epistasis analysis defined several pathways involving these proteins. Csm3 and Tof1 function in a same pathway and downstream of H2A. In addition, the pathway involving H2A/Csm3/Tof1 is distinct from the pathways involving the Ctf8/Ctf18/Dcc1 complex, the Rad9 pathway, and another involving Mrc1. Our genetic studies suggest a role for H2A serine 129 in the establishment of specialized cohesion structure necessary for the normal repair of topoisomerase I-induced DNA damage. PMID- 16219778 TI - Lysis timing and bacteriophage fitness. AB - The effect of lysis timing on bacteriophage (phage) fitness has received little theoretical or experimental attention. Previously, the impact of lysis timing on phage fitness was studied using a theoretical model based on the marginal value theorem from the optimal foraging theory. An implicit conclusion of the model is that, for any combination of host quantity and quality, an optimal time to lyse the host would exist so that the phage fitness would be the highest. To test the prediction, an array of isogenic lambda-phages that differ only in their lysis times was constructed. For each phage strain, the lysis time, burst size, and fitness (growth rate) were determined. The result showed that there is a positive linear relationship between lysis time and burst size. Moreover, the strain with an intermediate lysis time has the highest fitness, indicating the existence of an optimal lysis time. A mathematical model is also constructed to describe the population dynamics of phage infection. Computer simulations using parameter values derived from phage lambda-infection also showed an optimal lysis time. However, both the optimum and the fitness are different from the experimental result. The evolution of phage lysis timing is discussed from the perspectives of multiple infection and life-history trait evolution. PMID- 16219779 TI - Effect of mutation of the tetratricopeptide repeat and asparatate-proline 2 domains of Sti1 on Hsp90 signaling and interaction in Saccharomyces cerevisiae. AB - Through simultaneous interactions with Hsp70 and Hsp90 via separate tetratricopeptide repeat (TPR) domains, the cochaperone protein Hop/Sti1 has been proposed to play a critical role in the transfer of client proteins from Hsp70 to Hsp90. However, no prior mutational analysis demonstrating a critical in vivo role for the TPR domains of Sti1 has been reported. We used site-directed mutagenesis of the TPR domains combined with a genetic screen to isolate mutations that disrupt Sti1 function. A single amino acid alteration in TPR2A disrupted Hsp90 interaction in vivo but did not significantly affect function. However, deletion of a conserved residue in TPR2A or mutations in the carboxy terminal DP2 domain completely disrupted Sti1 function. Surprisingly, mutations in TPR1, previously shown to interact with Hsp70, were not sufficient to disrupt in vivo functions unless combined with mutations in TPR2B, suggesting that TPR1 and TPR2B have redundant or overlapping in vivo functions. We further examined the genetic and physical interaction of Sti1 with a mutant form of Hsp90, providing insight into the importance of the TPR2A domain of Sti1 in regulating Hsp90 function. PMID- 16219780 TI - No patrigenes required for femaleness in the haplodiploid wasp Nasonia vitripennis. AB - The parasitoid wasp Nasonia vitripennis is an emerging model organism for developmental and behavioral genetics. It reproduces by haplodiploidy; males typically develop parthenogenetically from haploid eggs and females from fertilized diploid eggs. A polyploid mutant strain is available in which females are triploid and lay haploid and diploid eggs that normally develop into males when unfertilized. In contrast to previous reports, approximately 2% of triploid females were found to occasionally produce daughters as well as gynandromorphs from diploid unfertilized eggs. Daughter production increased with age and differed among familial lineages. This is the first report of parthenogenetic female development in Nasonia. The results show that a paternally provided genome is not required for femaleness and call for modifications of existing models of sex determination in Nasonia. PMID- 16219781 TI - Drosophila target of rapamycin kinase functions as a multimer. AB - Target of rapamycin (TOR) is a conserved regulator of cell growth and metabolism that integrates energy, growth factor, and nutrient signals. The 280-kDa TOR protein functions as the catalytic component of two large multiprotein complexes and consists of an N-terminal HEAT-repeat domain and a C-terminal Ser/Thr kinase domain. Here we describe an allelic series of mutations in the Drosophila Tor gene and show that combinations of mutations in the HEAT and kinase domains of TOR display the rare genetic phenomenon of intragenic complementation, in which two or more defective proteins assemble to form a functional multimer. We present biochemical evidence that TOR self-associates in vivo and show that this multimerization is unaffected by positive or negative signals upstream of TOR. Consistent with multimerization of TOR, recessive mutations in the HEAT and kinase domains can dominantly interfere with wild-type TOR function in cells lacking TSC1 or TSC2. TOR multimerization thus partially accounts for the high apparent molecular weight of TOR complexes and offers novel therapeutic strategies for pathologies stemming from TOR hyperactivity. PMID- 16219782 TI - The mop1 (mediator of paramutation1) mutant progressively reactivates one of the two genes encoded by the MuDR transposon in maize. AB - Transposons make up a sizable portion of most genomes, and most organisms have evolved mechanisms to silence them. In maize, silencing of the Mutator family of transposons is associated with methylation of the terminal inverted repeats (TIRs) surrounding the autonomous element and loss of mudrA expression (the transposase) as well as mudrB (a gene involved in insertional activity). We have previously reported that a mutation that suppresses paramutation in maize, mop1, also hypomethylates Mu1 elements and restores somatic activity to silenced MuDR elements. Here, we describe the progressive reactivation of silenced mudrA after several generations in a mop1 background. In mop1 mutants, the TIRA becomes hypomethylated immediately, but mudrA expression and significant somatic reactivation is not observed until silenced MuDR has been exposed to mop1 for several generations. In subsequent generations, individuals that are heterozygous or wild type for the Mop1 allele continue to exhibit hypomethylation at Mu1 and mudrA TIRs as well as somatic activity and high levels of mudrA expression. Thus, mudrA silencing can be progressively and heritably reversed. Conversely, mudrB expression is never restored, its TIR remains methylated, and new insertions of Mu elements are not observed. These data suggest that mudrA and mudrB silencing may be maintained via distinct mechanisms. PMID- 16219783 TI - Intragenic suppression of Gal3C interaction with Gal80 in the Saccharomyces cerevisiae GAL gene switch. AB - Gal4-mediated activation of GAL gene transcription in Saccharomyces cerevisiae requires the interaction of Gal3 with Gal80, the Gal4 inhibitor protein. While it is known that galactose and ATP activates Gal3 interaction with Gal80, neither the mechanism of activation nor the surface that binds to Gal80 is known. We addressed this through intragenic suppression of GAL3C alleles that cause galactose-independent Gal3-Gal80 interaction. We created a new allele, GAL3SOC, and showed that it suppressed a new GAL3C allele. We tested the effect of GAL3SOC on several newly isolated and existing GAL3C alleles that map throughout the gene. All except one GAL3C allele, D368V, were suppressible by GAL3SOC. GAL3SOC and all GAL3C alleles were localized on a Gal3 homology model that is based on the structure of the highly related Gal1 protein. These results provide evidence for allosterism in the galactose- and ATP-activation of Gal3 binding to Gal80. In addition, because D368V and residues corresponding to Gal80-nonbinder mutations colocalized to a domain that is absent in homologous proteins that do not bind to Gal80, we suggest that D368 is a part of the Gal80-binding surface. PMID- 16219784 TI - Variation in mutation dynamics across the maize genome as a function of regional and flanking base composition. AB - We examine variation in mutation dynamics across a single genome (Zea mays ssp. mays) in relation to regional and flanking base composition using a data set of 10,472 SNPs generated by resequencing 1776 transcribed regions. We report several relationships between flanking base composition and mutation pattern. The A + T content of the two sites immediately flanking the mutation site is correlated with rate, transition bias, and GC --> AT pressure. We also observe a significant CpG effect, or increase in transition rate at CpG sites. At the regional level we find that the strength of the CpG effect is correlated with regional A + T content, ranging from a 1.7-fold increase in transition rate in relatively G + C rich regions to a 2.6-fold increase in A + T-rich regions. We also observe a relationship between locus A + T content and GC --> AT pressure. This regional effect is in opposition to the influence of the two immediate neighbors in that GC --> AT pressure increases with increasing locus A + T content but decreases with increasing flanking base A + T content and may represent a relationship between genome location and mutation bias. The data indicate multiple context effects on mutations, resulting in significant variation in mutation dynamics across the genome. PMID- 16219786 TI - Accumulation of nonfunctional S-haplotypes results in the breakdown of gametophytic self-incompatibility in tetraploid Prunus. AB - The transition from self-incompatibility (SI) to self-compatibility (SC) is regarded as one of the most prevalent transitions in Angiosperm evolution, having profound impacts on the genetic structure of populations. Yet, the identity and function of mutations that result in the breakdown of SI in nature are not well understood. This work provides the first detailed genetic description of the breakdown of S-RNase-mediated gametophytic self-incompatibility (GSI) in a polyploid species that exhibits genotype-dependent loss of SI. Genetic analyses of six natural sour cherry (Rosaceae, Prunus cerasus) selections identified seven independent, nonfunctional S-haplotypes with disrupted pistil component (stylar S) and/or pollen component (pollen-S) function. A genetic model demonstrating that the breakdown of SI in sour cherry is due to the accumulation of a minimum of two nonfunctional S-haplotypes within a single individual is developed and validated. Our finding that sour cherry is SI when only one nonfunctional S haplotype is present has significant evolutionary implications since nonfunctional S-haplotypes would be maintained in the population without causing an abrupt shift to SC. Furthermore, we demonstrate that heteroallelic sour cherry pollen is self-incompatible, which is counter to the well-documented phenomenon in the Solanaceae where SC accompanying polyploidization is frequently due to the SC of heteroallelic pollen. PMID- 16219785 TI - The twisted gene encodes Drosophila protein O-mannosyltransferase 2 and genetically interacts with the rotated abdomen gene encoding Drosophila protein O mannosyltransferase 1. AB - The family of mammalian O-mannosyltransferases includes two enzymes, POMT1 and POMT2, which are thought to be essential for muscle and neural development. Similar to mammalian organisms, Drosophila has two O-mannosyltransferase genes, rotated abdomen (rt) and DmPOMT2, encoding proteins with high homology to their mammalian counterparts. The previously reported mutant phenotype of the rt gene includes a clockwise rotation of the abdomen and defects in embryonic muscle development. No mutants have been described so far for the DmPOMT2 locus. In this study, we determined that the mutation in the twisted (tw) locus, tw(1), corresponds to a DmPOMT2 mutant. The twisted alleles represent a complementation group of recessive mutations that, similar to the rt mutants, exhibit a clockwise abdomen rotation phenotype. Several tw alleles were isolated in the past; however, none of them was molecularly characterized. We used an expression rescue approach to confirm that tw locus represents DmPOMT2 gene. We found that the tw1 allele represents an amino acid substitution within the conserved PMT domain of DmPOMT2 (TW) protein. Immunostaining experiments revealed that the protein products of both rt and tw genes colocalize within Drosophila cells where they reside in the ER subcellular compartment. In situ hybridization analysis showed that both genes have essentially overlapping patterns of expression throughout most of embryogenesis (stages 8-17), while only the rt transcript is present at early embryonic stages (5 and 6), suggesting its maternal origin. Finally, we analyzed the genetic interactions between rt and tw using several mutant alleles, RNAi, and ectopic expression approaches. Our data suggest that the two Drosophila O-mannosyltransferase genes, rt and tw, have nonredundant functions within the same developmental cascade and that their activities are required simultaneously for possibly the same biochemical process. Our results establish the possibility of using Drosophila as a model system for studying molecular and genetic mechanisms of protein O-mannosylation during development. PMID- 16219787 TI - DNA polymerase 4 of Saccharomyces cerevisiae is important for accurate repair of methyl-methanesulfonate-induced DNA damage. AB - The DNA polymerase 4 protein (Pol4) of Saccharomyces cerevisiae is a member of the X family of DNA polymerases whose closest human relative appears to be DNA polymerase lambda. Results from previous genetic studies conflict over the role of Pol4 in vivo. Here we show that deletion of Pol4 in a diploid strain of the SK1 genetic background results in sensitivity to methyl methanesulfonate (MMS). However, deletion of Pol4 in other strain backgrounds and in haploid strains does not yield an observable phenotype. The MMS sensitivity of a Pol4-deficient strain can be rescued by deletion of YKu70. We also show that deletion of Pol4 results in a 6- to 14-fold increase in the MMS-induced mutation frequency and in a significant increase in AT-to-TA transversions. Our studies suggest that Pol4 is critical for accurate repair of DNA lesions induced by MMS. PMID- 16219788 TI - Directional positive selection on an allele of arbitrary dominance. AB - Most models of positive directional selection assume codominance of the beneficial allele. We examine the importance of this assumption by implementing a coalescent model of positive directional selection with arbitrary dominance. We find that, for a given mean fixation time, a beneficial allele has a much weaker effect on diversity at linked neutral sites when the allele is recessive. PMID- 16219789 TI - Unequal contribution of sexes in the origin of dog breeds. AB - Dogs (Canis familiaris) were domesticated from the gray wolf (Canis lupus) at least 14,000 years ago, and there is evidence of dogs with phenotypes similar to those in modern breeds 4000 years ago. However, recent genetic analyses have suggested that modern dog breeds have a much more recent origin, probably <200 years ago. To study the origin of contemporaneous breeds we combined the analysis of paternally inherited Y chromosome markers with maternally inherited mitochondrial DNA and biparentally inherited autosomal microsatellite markers in both domestic dogs and their wild ancestor, the gray wolf. Our results show a sex bias in the origin of breeds, with fewer males than females contributing genetically, which clearly differs from the breeding patterns in wild gray wolf populations where both sexes have similar contributions. Furthermore, a comparison of mitochondrial DNA and Y chromosome diversity in dog groups recognized by the World Canine Organization, as well as in groups defined by the breeds' genetic composition, shows that paternal lineages are more differentiated among groups than maternal lineages. This demonstrates a lower exchange of males than of females between breeds belonging to different groups, which illustrates how breed founders may have been chosen. PMID- 16219790 TI - Growth defect and mutator phenotypes of RecQ-deficient Neurospora crassa mutants separately result from homologous recombination and nonhomologous end joining during repair of DNA double-strand breaks. AB - RecQ helicases function in the maintenance of genome stability in many organisms. The filamentous fungus Neurospora crassa has two RecQ homologs, QDE3 and RECQ2. We found that the qde-3 recQ2 double mutant showed a severe growth defect. The growth defect was alleviated by mutation in mei-3, the homolog of yeast RAD51, which is required for homologous recombination (HR), suggesting that HR is responsible for this phenotype. We also found that the qde-3 recQ2 double mutant showed a mutator phenotype, yielding mostly deletions. This phenotype was completely suppressed by mutation of mus-52, a homolog of the human KU80 gene that is required for nonhomologous end joining (NHEJ), but was unaffected by mutation of mei-3. The high spontaneous mutation frequency in the double mutant is thus likely to be due to NHEJ acting on an elevated frequency of double-strand breaks (DSBs) and we therefore suggest that QDE3 and RECQ2 maintain chromosome stability by suppressing the formation of spontaneous DSBs. PMID- 16219792 TI - Genetic evidence that nonhomologous disjunction and meiotic drive are properties of wild-type Drosophila melanogaster male meiosis. AB - We have followed sex and second chromosome disjunction, and the effects of these chromosomes on sperm function, in four genotypes: wild-type males, males deficient for the Y-linked crystal locus, males with an X chromosome heterochromatic deficiency that deletes all X-Y pairing sites, and males with both deficiencies. Both mutant situations provoke chromosome misbehavior, but the disjunctional defects are quite different. Deficiency of the X heterochromatin, consonant with the lack of pairing sites, mostly disrupts X-Y disjunction with a decidedly second-level effect on major autosome behavior. Deleting crystal, consonant with the cytological picture of postpairing chromatin-condensation problems, disrupts sex and autosome disjunction equally. Even when the mutant induced nondisjunction has very different mechanics, however, and even more importantly, even in the wild type, there is strong, and similar, meiotic drive. The presence of meiotic drive when disjunction is disrupted by distinctly different mechanisms supports the notion that drive is a normal cellular response to meiotic problems rather than a direct effect of particular mutants. Most surprisingly, in both wild-type and crystal-deficient males the Y chromosome moves to the opposite pole from a pair of nondisjoined second chromosomes nearly 100% of the time. This nonhomologous interaction is, however, absent when the X heterochromatin is deleted. The nonhomologous disjunction of the sex and second chromosomes may be the genetic consequence of the chromosomal compartmentalization seen by deconvolution microscopy, and the absence of Y-2 disjunction when the X heterochromatin is deleted suggests that XY pairing itself, or a previously unrecognized heterochromatic function, is prerequisite to this macrostructural organization of the chromosomes. PMID- 16219794 TI - Down-regulating constitutive activation of the NF-kappaB canonical pathway overcomes the resistance of cutaneous T-cell lymphoma to apoptosis. AB - Constitutive activation of the nuclear factor-kappaB (NF-kappaB) pathway has been shown to be involved in the resistance of tumor cells to apoptosis in several human malignancies of the hematopoietic lineage. By using electrophoretic mobility shift assay (EMSA) and confocal microscopic analysis, we demonstrate that NF-kappaB is constitutively activated in cutaneous T-cell lymphoma (CTCL) cell lines HuT-78, MyLa, and SeAx and in peripheral blood lymphocytes (PBLs) from patients with Sezary syndrome (SS) presenting a high ratio of tumor cells, with evidence of p50 and RelA/p65 in DNA-linked complexes. Transfection of SeAx line with a kappaB/luciferase reporter plasmid showed that translocated NF-kappaB complexes were functional. Selective inhibition of NF-kappaB, by transfecting CTCL cell lines with a super-repressor form of IkappaB alpha, led to apoptosis. We evidenced down-regulation of NF-kappaB activation and induction of CTCL cell apoptosis in the presence of proteasome 26S inhibitors ALLN, MG132, and bortezomib. Bortezomib at nanomolar concentrations inhibited constitutive activation of NF-kappaB and induced apoptosis of CTCL cells, with evidence of an upregulation of Bax expression. These results demonstrate the key role played by NF-kappaB in the resistance of CTCL to apoptosis and suggest that bortezomib might be useful for the treatment of patients with advanced stages of CTCL refractory to standard antineoplastic chemotherapy. PMID- 16219795 TI - Interferon regulatory factor 3 is involved in Toll-like receptor 4 (TLR4)- and TLR3-induced IL-12p35 gene activation. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine produced by dendritic cells (DCs) in response to Toll-like receptor (TLR) ligation. While the mechanisms regulating IL-12p40 chain gene expression are well characterized, molecular events involved in IL-12p35 chain gene activation remain to be clarified. Since IL-12p35 mRNA was induced in human DCs activated through TLR3 or TLR4 but not TLR2, we investigated the potential role of interferon regulatory factor 3 (IRF 3) in IL-12p35 gene transactivation. First, a binding site for IRF-3 named interferon-stimulated response element-1 (ISRE-1) was identified in the human IL 12p35 promoter region between nucleotides -251 and -240. The ISRE-1 site was required for IL-12p35 gene activation in RAW 264.7 cells stimulated by lipopolysaccharide (LPS) or PolyI:C. Ectopic expression of IRF-3 was found to up regulate IL-12p35 gene activation in the same system. Furthermore, chromatin immunoprecipitation (ChIP) studies demonstrated that IRF-3 is recruited to ISRE-1 site in TLR4- or TLR3-stimulated human DCs. Finally, experiments on DCs from IRF 3-deficient mice established that TLR4-induced IL-12p35 mRNA and IL-12p70 synthesis are impaired in absence of IRF-3. We conclude that IRF-3 binds to a critical cis-acting element in the IL-12p35 gene promoter and thereby represents a key factor for the induction of IL-12p70 synthesis in DCs. PMID- 16219791 TI - Extreme population-dependent linkage disequilibrium detected in an inbreeding plant species, Hordeum vulgare. AB - In human genetics a detailed knowledge of linkage disequilibrium (LD) is considered a prerequisite for effective population-based, high-resolution gene mapping and cloning. Similar opportunities exist for plants; however, differences in breeding system and population history need to be considered. Here we report a detailed study of localized LD in different populations of an inbreeding crop species. We measured LD between and within four gene loci within the region surrounding the hardness locus in three different gene pools of barley (Hordeum vulgare). We demonstrate that LD extends to at least 212 kb in elite barley cultivars but is rapidly eroded in related inbreeding ancestral populations. Our results indicate that haplotype-based sequence analysis in multiple populations will provide new opportunities to adjust the resolution of association studies in inbreeding crop species. PMID- 16219793 TI - The CRAL/TRIO and GOLD domain protein CGR-1 promotes induction of vulval cell fates in Caenorhabditis elegans and interacts genetically with the Ras signaling pathway. AB - Ras-mediated signaling is necessary for the induction of vulval cell fates during Caenorhabditis elegans development. We identified cgr-1 by screening for suppressors of the ectopic vulval cell fates caused by a gain-of-function mutation of the let-60 ras gene. Analysis of two cgr-1 loss-of-function mutations indicates that cgr-1 positively regulates induction of vulval cell fates. cgr-1 is likely to function at a step in the Ras signaling pathway that is downstream of let-60, which encodes Ras, and upstream of lin-1, which encodes a transcription factor, if these genes function in a linear signaling pathway. These genetic studies are also consistent with the model that cgr-1 functions in a parallel pathway that promotes vulval cell fates. Localized expression studies suggest that cgr-1 functions cell autonomously to affect vulval cell fates. cgr-1 also functions early in development, since cgr-1 is necessary for larval viability. CGR-1 contains a CRAL/TRIO domain likely to bind a small hydrophobic ligand and a GOLD domain that may mediate interactions with proteins. A bioinformatic analysis revealed that there is a conserved family of CRAL/TRIO and GOLD domain-containing proteins that includes members from vertebrates and Drosophila. The analysis of cgr-1 identifies a novel in vivo function for a member of this family and a potential new regulator of Ras-mediated signaling. PMID- 16219796 TI - Laminin stimulates spreading of platelets through integrin alpha6beta1-dependent activation of GPVI. AB - The extracellular matrix protein, laminin, supports platelet adhesion through binding to integrin alpha6beta1 In the present study, we demonstrate that human laminin, purified from placenta, also stimulates formation of filopodia and lamellipodia in human and mouse platelets through a pathway that is dependent on alpha6beta1 and the collagen receptor GPVI. The integrin alpha6beta1 is essential for adhesion to laminin, as demonstrated using an alpha6-blocking antibody, whereas GPVI is dispensable for this response, as shown using "knockout" mouse platelets. On the other hand, lamellipodia formation on laminin is completely inhibited in the absence of GPVI, although filopodia formation remains and is presumably mediated via alpha6beta1 Lamellipodia and filopodia formation are inhibited in Syk-deficient platelets, demonstrating a key role for the kinase in signaling downstream of GPVI and integrin alpha6beta1 GPVI was confirmed as a receptor for laminin using surface plasmon resonance spectroscopy and by demonstration of lamellipodia formation on laminin in the presence of collagenase. These results identify GPVI as a novel receptor for laminin and support a model in which integrin alpha6beta1 brings laminin to GPVI, which in turn mediates lamellipodia formation. We speculate that laminin contributes to platelet spreading in vivo through a direct interaction with GPVI. PMID- 16219797 TI - Fludarabine plus cyclophosphamide versus fludarabine alone in first-line therapy of younger patients with chronic lymphocytic leukemia. AB - Combination chemotherapy with fludarabine plus cyclophosphamide (FC) was compared with the standard regimen of fludarabine monotherapy in first-line treatment of younger patients with chronic lymphocytic leukemia (CLL). Between 1999 and 2003, a total of 375 patients younger than 66 years who predominantly had advanced CLL were randomly assigned to receive either fludarabine (25 mg/m(2) for 5 days intravenously, repeated every 28 days) or FC combination therapy (fludarabine 30 mg/m(2) plus cyclophosphamide 250 mg/m(2) for 3 days intravenously, repeated every 28 days). Both regimens were administered to a maximum of 6 courses. FC combination chemotherapy resulted in significantly higher complete remission rate (24%) and overall response rate (94%) compared with fludarabine alone (7% and 83%; P < .001 and P = .001). FC treatment also resulted in longer median progression-free survival (48 vs 20 months; P = .001) and longer treatment-free survival (37 vs 25 months; P < .001). Thus far, no difference in median overall survival has been observed. FC caused significantly more thrombocytopenia and leukocytopenia but did not increase the number of severe infections. In summary, first-line treatment with FC increases the response rates and the treatment-free interval in younger patients with advanced CLL. PMID- 16219798 TI - SLAM family markers are conserved among hematopoietic stem cells from old and reconstituted mice and markedly increase their purity. AB - Recent advances have increased the purity of hematopoietic stem cells (HSCs) isolated from young mouse bone marrow. However, little attention has been paid to the purity of HSCs from other contexts. Although Thy-1 low Sca-1+ Lineage- c-kit+ cells from young bone marrow are highly enriched for HSCs (1 in 5 cells gives long-term multilineage reconstitution after transplantation into irradiated mice), the same population from old, reconstituted, or cytokine-mobilized mice engrafts much less efficiently (1 in 78 to 1 in 185 cells gives long-term multilineage reconstitution). To test whether we could increase the purity of HSCs isolated from these contexts, we examined the SLAM family markers CD150 and CD48. All detectable HSCs from old, reconstituted, and cyclophosphamide/G-CSF mobilized mice were CD150+ CD48-, just as in normal young bone marrow. Thy-1 low Sca-1+ Lineage- c-kit+ cells from old, reconstituted, or mobilized mice included mainly CD48+ and/or CD150- cells that lacked reconstituting ability. CD150+ CD48- Sca-1+ Lineage- c-kit+ cells from old, reconstituted, or mobilized mice were much more highly enriched for HSCs, with 1 in 3 to 1 in 7 cells giving long-term multilineage reconstitution. SLAM family receptor expression is conserved among HSCs from diverse contexts, and HSCs from old, reconstituted, and mobilized mice engraft relatively efficiently after transplantation when contaminating cells are eliminated. PMID- 16219799 TI - Loss of Tie2 receptor compromises embryonic stem cell-derived endothelial but not hematopoietic cell survival. AB - Tie2 is a receptor-type tyrosine kinase expressed on hematopoietic stem cells and endothelial cells. We used cultured embryonic stem (ES) cells to determine the function of Tie2 during early vascular development and hematopoiesis. Upon differentiation, the ES cell-derived Tie2+ Flk1+ fraction was enriched for hematopoietic and endothelial progenitor cells. To investigate lymphatic differentiation, we used a monoclonal antibody against LYVE-1 and found that LYVE 1+ cells derived from Tie2+ Flk1+ cells possessed various characteristics of lymphatic endothelial cells. To determine whether Tie2 played a role in this process, we analyzed differentiation of Tie2-/- ES cells. Although the initial numbers of LYVE-1+ and PECAM-1+ cells derived from Tie2-/- cells did not vary significantly, the number of both decreased dramatically upon extended culturing. Such decreases were rescued by treatment with a caspase inhibitor, suggesting that reductions were due to apoptosis as a consequence of a lack of Tie2 signaling. Interestingly, Tie2-/- ES cells did not show measurable defects in development of the hematopoietic system, suggesting that Tie2 is not essential for hematopoietic cell development. PMID- 16219800 TI - Stem cell transplantation with reduced-intensity conditioning for hemophagocytic lymphohistiocytosis. AB - Allogeneic stem cell transplantation (SCT) is curative for hemophagocytic lymphohistiocytosis (HLH). However, patients frequently have significant morbidity before transplantation and there is high transplant-related mortality (TRM). Because first-degree HLH is caused by immune dysregulation, a reduced intensity conditioned (RIC) regimen might be sufficient for cure while decreasing the TRM. Twelve patients with HLH underwent RIC SCT from a matched family/unrelated or haploidentical donor. Eleven were conditioned with fludarabine/melphalan with additional busulphan for haploidentical grafts. One received fludarabine and 2-Gy total body irradiation (TBI). All patients showed engraftment at a median of 14 days. Nine of 12 (75%) are alive and in complete remission (CR) a median of 30 months (range, 9-73 months) after SCT. Two patients died from pneumonitis and one from hepatic rupture. Four patients developed acute graft-versus-host disease (GVHD) and 3 have chronic GVHD. Three of 9 survivors have mixed chimerism but remain free of disease. In summary, RIC compares favorably to conventional SCT with long-term disease control in surviving patients despite a significant incidence of mixed chimerism. PMID- 16219801 TI - Virus-stimulated plasmacytoid dendritic cells induce CD4+ cytotoxic regulatory T cells. AB - Immune responses to pathogens need to be maintained within appropriate levels to minimize tissue damage, whereas such controlled immunity may allow persistent infection of certain types of pathogens. Interleukin 10 (IL-10) plays an important role in such immune regulation. We previously showed that HSV stimulated human plasmacytoid dendritic cells (pDCs) induced naive CD4+ T cells to differentiate into interferon gamma (IFN-gamma)/IL-10-producing T cells. Here we show that HSV-stimulated pDCs induce allogeneic naive CD4+ T cells to differentiate into cytotoxic regulatory T cells that poorly proliferate on restimulation and inhibit proliferation of coexisting naive CD4+ T cells. IL-3 stimulated pDCs or myeloid DCs did not induce such regulatory T cells. Both IFN alpha and IL-10 were responsible for the induction of anergic and regulatory properties. High percentages of CD4+ T cells cocultured with HSV-stimulated pDCs, and to a lesser extent those cocultured with IL-3-stimulated pDCs, expressed granzyme B and perforin in an IL-10-dependent manner. CD4+ T cells cocultured with HSV-stimulated pDCs accordingly exhibited cytotoxic activity. The finding that virus-stimulated pDCs are capable of inducing CD4+ cytotoxic regulatory T cells suggests that this DC subset may play an important role in suppressing excessive inflammatory responses and also in inducing persistent viral infection. PMID- 16219804 TI - CpG-containing oligodeoxynucleotide promotes microglial cell uptake of amyloid beta 1-42 peptide by up-regulating the expression of the G-protein- coupled receptor mFPR2. AB - Human G protein-coupled formyl peptide receptor like 1 (FPRL1) and its mouse homologue murine formyl peptide receptor 2 (mFPR2) mediate the chemotactic activity of amyloid beta 1-42 (Abeta42), a key pathogenic peptide in Alzheimer's disease (AD). Since mFPR2 is up-regulated in mouse microglia by lipopolysaccharide (LPS), a Toll-like receptor 4 ligand, we investigated the capacity of CpG-containing oligodeoxynucleotide (ODN), a Toll-like receptor (TLR) 9 ligand, to regulate the expression of mFPR2 in mouse microglia. CpG ODN markedly enhanced the expression and function of mFPR2 in microglial cells, which exhibited increased chemotactic responses to mFPR2 agonists, including Abeta42. The effect of CpG ODN is dependent on activation of p38 MAPK. Further studies showed that CpG ODN-treated microglia increased their capacity to endocytose Abeta42 through mFPR2, as this process was abrogated by pertussis toxin, a Gi protein inhibitor, and W peptide, another potent mFPR2 agonist. Our results suggest that TLR9 may play an important role in promoting microglial recognition of Abeta42, thus affecting the pathogenic process of AD. PMID- 16219805 TI - Up-regulation of CD36/FAT in preadipocytes in familial combined hyperlipidemia. AB - Familial combined hyperlipidemia (FCHL) shows many features of the metabolic syndrome. The strong genetic component makes it an excellent model to study the genetic background of metabolic syndrome and insulin resistance. Adipose tissue is believed to contribute to, or even underlie, the FCHL phenotype and is an interesting target tissue for gene expression studies. However, interpretation of adipose tissue gene expression experiments is complex since expression differences cannot only arise as a direct consequence of a genetic trait, but may also reflect an adaptation to metabolic influences at the cellular level. In the present study, we measured gene expression levels in cultured primary human preadipocytes from FCHL and control subjects. Since isolated preadipocytes were allowed to replicate for weeks under standardized conditions, the contribution of previous metabolic influences is rather small whereas genetic defects are preserved and expressed in vitro. The main finding was up-regulation of CD36/FAT in FCHL preadipocytes, confirmed in two independent groups of subjects, and a concomitant increase in CD36/FAT-mediated fatty acid uptake. CD36/FAT overexpression has previously been shown to be associated with other insulin resistant states. The present data suggest that CD36/FAT overexpression in FCHL occurs very early in adipocyte differentiation and may be of genetic origin. PMID- 16219802 TI - Up-regulation of the Notch ligand Delta-like 4 inhibits VEGF-induced endothelial cell function. AB - Delta-like 4 (Dll4), a membrane-bound ligand for Notch1 and Notch4, is selectively expressed in the developing endothelium and in some tumor endothelium, and it is induced by vascular endothelial growth factor (VEGF)-A and hypoxia. Gene targeting studies have shown that Dll4 is required for normal embryonic vascular remodeling, but the mechanisms underlying Dll4 regulatory functions are currently not defined. In this study, we generated primary human endothelial cells that overexpress Dll4 protein to study Dll4 function and mechanism of action. Human umbilical vein endothelial cells retrovirally transduced with Dll4 displayed reduced proliferative and migratory responses selectively to VEGF-A. Expression of VEGF receptor-2, the principal signaling receptor for VEGF-A in endothelial cells, and coreceptor neuropilin-1 was significantly decreased in Dll4-transduced endothelial cells. Consistent with Dll4 signaling through Notch, expression of HEY2, one of the transcription factors that mediates Notch function, was significantly induced in Dll4 overexpressing endothelial cells. The gamma-secretase inhibitor L-685458 significantly reconstituted endothelial cell proliferation inhibited by immobilized extracellular Dll4 and reconstituted VEGFR2 expression in Dll4 overexpressing endothelial cells. These results identify the Notch ligand Dll4 as a selective inhibitor of VEGF-A biologic activities down-regulating 2 VEGF receptors expressed on endothelial cells and raise the possibility that Dll4 may be exploited therapeutically to modulate angiogenesis. PMID- 16219806 TI - Chemical synthesis and thermodynamic characterization of oxanine-containing oligodeoxynucleotides. AB - Oxanine (Oxa, O), one of the major damaged bases from guanine generated by NO- or HNO2-induced nitrosative deamination, has been considered as a mutagen-potent lesion. For exploring more detailed properties of Oxa, large-scale preparation of Oxa-containing oligodeoxynucleotide (Oxa-ODN) with the desired base sequence is a prerequisite. In the present study, we have developed a chemical synthesis procedure of Oxa-ODNs and characterized thermodynamic properties of Oxa in DNA strands. First, 2'-deoxynucleoside of Oxa (dOxo) obtained from 2'-deoxyguanosine by HNO2-nitrosation was subjected to 5'-O-selective tritylation to give 5'-O (4,4'-dimethoxytrityl)-dOxo (DMT-dOxo) with a maximum yield of 70%. Subsequently, DMT-dOxo was treated with conventional phosphoramidation, which resulted in DMT dOxo-amidite monomer with a maximum yield of 72.5%. The amidite obtained was used for synthesizing Oxa-ODNs: the coupling yields for Oxa incorporation were over 93%. The prepared Oxa-ODNs were employed for analyzing the thermodynamic properties of DNA duplexes containing base-matches of O:N [N; C (cytosine), T (thymine), G (guanine) or A (adenine)]. Melting temperatures (Tm) and thermodynamic stability (DeltaG37(0)) were found to be lower by 6.83 approximately 13.41 degrees C and 2.643 approximately 6.047 kcal mol(-1), respectively, compared with those of oligodeoxynucleotides, which had the same base sequence except that O:N was replaced by G:C (wild type). It has also been found that Oxa-pairing with cytosine shows relatively high stability in DNA duplex compared with other base combinations. The orders of DeltaDeltaG37(0) were O:C > O:T > O:A > O:G. The chemical synthesis procedure and thermodynamic characteristics of Oxa-ODNs established here will be helpful for elucidating the biological significance of Oxa in relation to genotoxic and repair mechanisms. PMID- 16219803 TI - Regulation of LIM-kinase 1 and cofilin in thrombin-stimulated platelets. AB - Cofilin is a regulator of actin filament dynamics. We studied whether during platelet activation Rho kinase stimulates LIM kinase (LIMK) leading to subsequent phosphorylation and inactivation of cofilin. Platelet shape change and aggregation/secretion were induced by low and high concentrations of thrombin, respectively. We found that during these platelet responses Rho kinase activation was responsible for mediating rapid Thr508 phosphorylation and activation of LIMK 1 and for the F-actin increase during shape change and, in part, during secretion. Surprisingly, during shape change cofilin phosphorylation was unaltered, and during aggregation/secretion cofilin was first rapidly dephosphorylated by an okadaic acid-insensitive phosphatase and then slowly rephosphorylated by LIMK-1. LIMK-1 phosphorylation and cofilin dephosphorylation and rephosphorylation during aggregation were independent of integrin alpha(IIb)beta(3) engagement. Cofilin phosphorylation did not regulate cofilin association with F-actin and was unrelated to the F-actin increase in thrombin activated platelets. Our study identifies LIMK-1 as being activated by Rho kinase in thrombin-stimulated platelets. Two counteracting pathways, a cofilin phosphatase and LIMK-1, are activated during platelet aggregation/secretion regulating cofilin phosphorylation sequentially and independently of integrin alpha(IIb)beta(3) engagement. Rho kinase-mediated F-actin increase during platelet shape change and secretion involves a mechanism other than LIMK-1 mediated cofilin phosphorylation, raising the possibility of another LIMK substrate regulating platelet actin assembly. PMID- 16219807 TI - Digitalis: new actions for an old drug. AB - The mechanisms by which digitalis causes its therapeutic and toxic actions have been studied for nearly a half century, revealing a great deal about cardiac cell regulation of intracellular ions via the Na-K-ATPase (NKA) and how it is altered by cardiac glycosides. However, recent observations suggest that digitalis may have additional effects on cardiac cell function in both the short and long term that include intracellular effects, interactions with specific NKA isoforms in different cellular locations, effects on intracellular (including nuclear) signaling, and long-term regulation of intracellular ionic balances through circulating ouabain-like compounds. The purpose of this review is to examine the current status of a number of the newest and most interesting developments in the study of digitalis with a particular focus on cardiac function, although we will also discuss some of the new advances in other relevant cardiovascular effects. This new information has important implications for both our understanding of ionic regulation in normal and diseased hearts as well as for potential avenues for the development of future therapeutic interventions for the treatment of heart failure. PMID- 16219808 TI - Is there a pathophysiological role for perivascular adipocytes? PMID- 16219809 TI - Novel mechanism of action of ACE and its inhibitors. PMID- 16219810 TI - Angiotensin-converting enzyme regulates bradykinin receptor gene expression. AB - The angiotensin-converting enzyme (ACE) is a membrane-bound peptidyl dipeptidase known to act on a variety of peptide substrates in the extracellular space. Its most notable functions are the formation of angiotensin II and the degradation of bradykinin. In the current experiments, we found that exogenous ACE added to vascular smooth muscle cell culture strongly induces and upregulates the genes of bradykinin receptors B1 and B2. This transcriptional regulatory property of ACE was shown to be unrelated to its known enzymatic properties. Indeed, ACE at 3.75 microg/ml added in the culture medium of vascular smooth muscle cells was found to cause marked upregulation of the mRNA expression of the genes for the B1 and B2 receptors of bradykinin by 22- and 11-fold, respectively. This phenomenon was not altered by the addition of specific angiotensin II antagonists for the AT1 or AT2 receptors. Moreover, the ACE inhibitor captopril, which inhibited ACE enzymatic activity, did not block its effect at the bradykinin receptor gene transcription level. Expression of both receptor genes was completely abolished by actinomycin D. Furthermore, transcriptional upregulation was inhibited by curcumin, suggesting involvement of different transcriptional factors in this phenomenon. Electrophoretic mobility shift assay revealed increase in NF-kappaB and activator protein-1 protein binding for consensus sequences, between ACE treated cells versus untreated cells. The data indicate a novel biological function of the ACE unrelated to its well-known enzymatic function as a peptidyl dipeptidase. PMID- 16219811 TI - Ex vivo electroporation as a potent new strategy for nonviral gene transfer into autologous vein grafts. AB - Gene transfer to vein grafts has therapeutic potential to prevent late graft failure; however, certain issues, including efficacy and safety, have hindered the clinical application of this treatment modality. Here, we report the successful and efficient gene transfer of plasmid DNA via ex vivo electroporation into veins as well as into vein grafts. Two approaches were used: one involved transluminal in situ gene transfer using a T-shaped electrode (the "Lu" method), and the other was an adventitial ex vivo approach using an electroporation cuvette followed by vein grafting (the "Ad" method). The Lu method was carried out at 10 V, with optimal gene transfer efficiency in the in situ jugular veins of rabbits, and transgene expression was observed primarily in endothelial cells. However, when these veins were grafted into the arterial circulation, no luciferase activity was detected; this effect was probably due to the elimination of the gene-transferred cells as a result of endothelial denudation. In contrast, optimal and satisfactory gene transfer was obtained with the vein grafts subjected to the Ad method at 30 V, and transgene expression was seen primarily in adventitial fibroblasts. Gene transfer of endothelial nitric oxide synthase cDNA to the vein graft via the Ad method successfully limited the extent of intimal hyperplasia, even under hyperlipidemic conditions, at 4 wk after grafting. We thus propose that the Ad method via ex vivo electroporation may provide a novel, safe, and clinically available technique for nonviral gene transfer to sufficiently prevent late graft failure. PMID- 16219812 TI - Regional ventricular wall thickening reflects changes in cardiac fiber and sheet structure during contraction: quantification with diffusion tensor MRI. AB - Dynamic changes of myocardial fiber and sheet structure are key determinants of regional ventricular function. However, quantitative characterization of the contraction-related changes in fiber and sheet structure has not been reported. The objective of this study was to quantify cardiac fiber and sheet structure at selected phases of the cardiac cycle. Diffusion tensor MRI was performed on isolated, perfused Sprague-Dawley rat hearts arrested or fixed in three states as follows: 1) potassium arrested (PA), which represents end diastole; 2) barium induced contracture with volume (BV+), which represents isovolumic contraction or early systole; and 3) barium-induced contracture without volume (BV-), which represents end systole. Myocardial fiber orientations at the base, midventricle, and apex were determined from the primary eigenvectors of the diffusion tensor. Sheet structure was determined from the secondary and tertiary eigenvectors at the same locations. We observed that the transmural distribution of the myofiber helix angle remained unchanged as contraction proceeded from PA to BV+, but endocardial and epicardial fibers became more longitudinally orientated in the BV group. Although sheet structure exhibited significant regional variations, changes in sheet structure during myocardial contraction were relatively uniform across regions. The magnitude of the sheet angle, which is an index of local sheet slope, decreased by 23 and 44% in BV+ and BV- groups, respectively, which suggests more radial orientation of the sheet. In summary, we have shown for the first time that geometric changes in both sheet and fiber orientation provide a substantial mechanism for radial wall thickening independent of active components due to myofiber shortening. Our results provide direct evidence that sheet reorientation is a primary determinant of myocardial wall thickening. PMID- 16219813 TI - Increasing donor age adversely impacts beneficial effects of bone marrow but not smooth muscle myocardial cell therapy. AB - We evaluated the impact of donor age on the efficacy of myocardial cellular therapy for ischemic cardiomyopathy. Characteristics of smooth muscle cells (SMC), bone marrow stromal cells (MSCs), and skeletal muscle cells (SKMCs) from young, adult, and old rats were compared in vitro. Three weeks after coronary ligation, 3.5 million SMCs (n = 11) or MSCs (n = 9) from old syngenic rats or culture medium (n = 6) were injected into the ischemic region. Five weeks after implantation, cardiac function was assessed by echocardiography and the Langendorff apparatus. In the in vitro study, the numbers and proliferation of MSCs from fresh bone marrow and SKMCs from fresh tissue but not SMCs were markedly diminished in old animals (P < 0.05 both groups). SKMCs from old animals did not reach confluence. After treatment with 5-azacytidine (azacitidine), the myogenic potential of old MSCs was decreased compared with young MSCs. In the in vivo study, both SMC and MSC transplantation induced significant angiogenesis compared with media injections (P < 0.05 both groups). Transplantation of SMCs but not MSCs prevented scar thinning (P = 0.03) and improved ejection fraction and fractional shortening (P < 0.05). Load-independent indices of cardiac function in a Langendorff preparation confirmed improved function in the aged SMC group (P = 0.01) but not in the MSC group compared with the control group. In conclusion, donor age adversely impacts the efficacy of cellular therapy for myocardial regeneration and is cell-type dependent. SMCs from old donors retain their ability to improve cardiac function after implantation into ischemic myocardium. PMID- 16219814 TI - Peroxynitrite hyperpolarizes smooth muscle and relaxes internal carotid artery in rabbit via ATP-sensitive K+ channels. AB - The goal of this study was to determine the effects of peroxynitrite (ONOO-) on smooth muscle membrane potential and vasomotor function in rabbit carotid arteries. ONOO- is known to affect vascular tone by several mechanisms, including effects on K+ channels. Xanthine (X, 0.1 mM), xanthine oxidase (XO, 0.01 U/ml), and a low concentration of sodium nitroprusside (SNP, 10 nM) were used to generate ONOO-. In the common carotid artery, X and XO (X/XO) in the presence of SNP tended to increase tension. In contrast, in the internal carotid artery, X/XO in the presence of SNP transiently hyperpolarized the membrane (-8.5 +/- 1.8 mV, mean +/- SE) and decreased tension (by 85 +/- 5.6%). In internal carotid arteries, in the absence of SNP, X/XO did not hyperpolarize the membrane and produced much less relaxation (by 23 +/- 5.6%) than X/XO and SNP. Ebselen (50 microM) inhibited both hyperpolarization and relaxation to X/XO and SNP, and uric acid (100 microM) inhibited relaxation. Glibenclamide (1 microM) abolished hyperpolarization and inhibited relaxation during X/XO and SNP. Charybdotoxin (100 nM) or tetraethylammonium (1 mM) did not affect hyperpolarization or relaxation, respectively. These results suggest that ONOO- hyperpolarizes and relaxes smooth muscle in rabbit internal carotid artery but not in common carotid artery through activation of K(ATP) channels. PMID- 16219815 TI - Caffeic acid phenethyl ester possesses potent cardioprotective effects in a rabbit model of acute myocardial ischemia-reperfusion injury. AB - Although great achievements have been made in elucidating the molecular mechanisms contributing to acute myocardial ischemia/reperfusion (I/R) injury, an effective pharmacological therapy to protect cardiac tissues from serious damage associated with acute myocardial infarction, coronary arterial bypass grafting surgery, or acute coronary syndromes has not been developed. We examined the in vivo cardioprotective effects of caffeic acid phenethyl ester (CAPE), a natural product with potent anti-inflammatory, antitumor, and antioxidant activities. CAPE was systemically delivered to rabbits either 60 min before or 30 min after surgically inducing I/R injury. Infarct dimensions in the area at risk were reduced by >2-fold (P < 0.01) with CAPE treatment at either period. Accordingly, serum levels of normally cytosolic enzymes lactate dehydrogenase, creatine kinase (CK), MB isoenzyme of CK, and cardiac-specific troponin I were markedly reduced in both CAPE treatment groups (P < 0.05) compared with the vehicle-treated control group. CAPE-treated tissues displayed significantly less cell death (P < 0.05), which was in part due to inhibition of p38 mitogen-activated protein kinase activation and reduced DNA fragmentation often associated with caspase 3 activation (P < 0.05). In addition, CAPE directly blocked calcium-induced cytochrome c release from mitochondria. Finally, the levels of inflammatory proteins IL-1beta and TNF-alpha expressed in the area at risk were significantly reduced with CAPE treatment (P < 0.05). These data demonstrate that CAPE has potent cardioprotective effects against I/R injury, which are mediated, at least in part, by the inhibition of inflammatory and cell death responses. Importantly, protection is conferred when CAPE is systemically administered after the onset of ischemia, thus demonstrating potential efficacy in the clinical scenario. PMID- 16219816 TI - Can MR spectroscopy ever be simple and effective? PMID- 16219817 TI - Diffusion tensor imaging at the crossroads: fiber tracking meets tissue characterization in brain tumors. PMID- 16219818 TI - Assessing global invasion of newly diagnosed glial tumors with whole-brain proton MR spectroscopy. AB - BACKGROUND AND PURPOSE: Because of their invasive nature, high-grade glial tumors are uniformly fatal. The purpose of this study was to quantify MR imaging-occult, glial tumor infiltration beyond its radiologic margin through its consequent neuronal cell damage, assessed by the global concentration decline of the neuronal marker N-acetylaspartate (NAA). METHODS: Seventeen patients (10 men; median age, 39 years; age range, 23-79 years) with radiologically suspected (later pathologically confirmed) supratentorial glial neoplasms, and 17 age- and sex-matched controls were studied. Their whole-brain NAA (WBNAA) amounts were obtained with proton MR spectroscopy: for patients on the day of surgery (n = 17), 1 day postsurgery (n = 15), and once for each control. To convert into concentrations, suitable for intersubject comparison, patients' global NAA amounts were divided by their brain volumes segmented from MR imaging. Least squares regression was used to analyze the data. RESULTS: Pre- and postoperative WBNAA (mean +/- SD) of 9.2 +/- 2.1 and 9.7 +/- 1.8 mmol/L, respectively, in patients were indistinguishable (P = .369) but significantly lower than in controls (12.5 +/- 1.4 mmol/L). Mean resected tumor size (n = 15) was approximately 3% of total brain volume. CONCLUSION: The average 26% WBNAA deficit in the patients, which persisted following surgical resection, cannot be explained merely by depletion within the approximately 3% MR imaging-visible tumor volume or an age-dependent effect. Although there could be several possible causes of such widespread decline--perineuronal satellitosis, neuronal deafferentation, Wallerian and retrograde degeneration, vasogenic edema, functional diaschisis, secondary vascular changes--most are a direct or indirect reflection of extensive, MR imaging-occult, microscopic tumor cell infiltration, diffusely throughout the otherwise "normal-appearing" brain. PMID- 16219819 TI - Brain compression without global neuronal loss in meningiomas: whole-brain proton MR spectroscopy report of 2 cases. AB - We report the findings from whole-brain proton MR spectroscopy, quantifying the neuronal marker N-acetylaspartate (NAA), for 2 presurgical meningioma patients and 10 healthy controls. The patients' whole-brain NAA (WBNAA) concentrations were considerably elevated (3+ SDs) compared with healthy controls when excluding the tumors from brain volume; WBNAA levels normalized following correction to approximate "preneoplastic" brain size. These results suggest global neuronal preservation in these 2 patients while their brains were compressed by large, slowly growing, extra-axial masses. PMID- 16219820 TI - Fiber density index correlates with reduced fractional anisotropy in white matter of patients with glioblastoma. AB - A quantitative description of white matter fiber attenuation, the fiber density index (FDi) is described. This measure is derived from quantitative analysis of diffusion tensor imaging and reveals an index of the number of fiber paths traversing a region of interest. In this report, FDi is computed in 10 patients with glioblastoma multiforme, in white matter that is peritumoral and contralateral. FDi is shown to correlate significantly with fractional anisotropy. PMID- 16219821 TI - Assessment of diagnostic accuracy of perfusion MR imaging in primary and metastatic solitary malignant brain tumors. AB - PURPOSE: The purpose of this study was to estimate the diagnostic accuracy of relative cerebral blood volume (rCBV) measurement in preoperative grading and differentiation of solitary intra-axial malignant brain tumors. METHODS: Thirty six low-grade glial tumors (LGGTs), 22 high-grade glial tumors (HGGTs), and 17 metastases (METs) were prospectively evaluated by MR imaging and standard dynamic susceptibility contrast-enhanced gradient echo, echoplanar imaging during first pass of a bolus injection of contrast material. Normalized rCBV values from tumoral (rCBV(T)) and peritumoral (rCBV(P)) areas were calculated by standard software and statistically tested independently. RESULTS: The mean differences of rCBV(T) and rCBV(P) values between LGGT (2.30 +/- 1.12 and 1.18 +/- 0.24) and HGGT (5.42 +/- 1.52 and 2.17 +/- 0.82) (P < .001); HGGTs and METs (3.21 +/- 0.98 and 0.97 +/- 0.09) (P < .001); and LGGTs and METs (P < .05 and P < .001, respectively) were significant. No clear cutoff value was present. A clear rCBV(T) cutoff value of 2.6 was detected for differentiation of low- (1.75 +/- 0.38; LGA) versus high-grade (4.78 +/- 0.99; HGA) astrocytomas when nonastrocytic glial tumors were excluded. The rCBV(T) values were linearly correlated with degree of malignancy (r = 0.869; P < .001). Cutoff rCBV(P) values of 1.1 and 1.2 were quite effective in differentiation of METs from LGGTs and HGGTs, respectively. The overall efficacy of rCBV was higher in grading than in differentiation. CONCLUSION: The diagnostic accuracy of rCBV measurement is higher in grading of glial brain tumors than in differentiation of HGGTs from solitary intra-axial METs. The astrocytic and nonastrocytic glial tumors have to be evaluated separately for precise grading. PMID- 16219822 TI - Evaluation of treatment-induced cerebral white matter injury by using diffusion tensor MR imaging: initial experience. AB - BACKGROUND AND PURPOSE: Treatment with chemotherapy and radiation therapy for brain tumors can cause white matter (WM) injury. Conventional MR imaging, however, cannot always depict treatment-induced transient WM abnormalities. We investigated the ability of diffusion-tensor (DT) MR imaging and proton MR spectroscopy to detect the treatment-induced transient changes within normal appearing WM. METHODS: DT MR imaging and proton MR spectroscopy were performed in 8 patients treated with a combination of surgery, chemotherapy, and radiation therapy for brain tumors (17 examinations) and 11 age-matched controls. Apparent diffusion coefficient (ADC) value, fractional anisotropy (FA) value, and N acetylaspartate (NAA)/creatine (Cr) ratio were obtained from 27 hemispheres with normal-appearing WM in the patients. We divided the datasets of isotropic ADC, FA, and NAA/Cr, on the basis of the time period after completion of radiation therapy, into 4 groups: group 1 (0-2 months; n = 10), group 2 (3-5 months; n = 5), group 3 (6-9 months; n = 7), and group 4 (10-12 months; n = 5). We compared averages of mean isotropic ADC, mean FA, and NAA/Cr of each patient group with those of the control group by using a t test. RESULTS: In the group 2, averages of mean FA and NAA/Cr decreased and average of mean isotopic ADC increased in comparison with those of the control group (P = .004, .04, and .0085, respectively). There were no significant differences in the averages between the control group and patient groups 1, 3, and 4. CONCLUSION: DT MR imaging and proton MR spectroscopy can provide quantitative indices that may reflect treatment-induced transient derangement of normal-appearing WM. PMID- 16219823 TI - Intracranial hemorrhage complicating acute stroke: how common is hemorrhagic stroke on initial head CT scan and how often is initial clinical diagnosis of acute stroke eventually confirmed? AB - BACKGROUND AND PURPOSE: Assessment of possible hemorrhage in acute stroke before appropriate therapy remains important. The aim of this study was to determine the frequency with which patients present with clinical stroke and have intracranial hemorrhage on initial noncontrast head CT scan (NCCT). In addition, we sought to determine the frequency with which initial clinical diagnosis acute stroke is confirmed in this group. METHODS: Medical records of 691 consecutive patients with admitting diagnosis of acute stroke were evaluated retrospectively. Results of initial NCCT performed within 24 hours after presentation were assessed. All patients were examined before anticoagulation or thrombolysis. Correlation with treatment and leading differential etiology was made. RESULTS: Twenty-five patients (25/691 [3.6%]) had hemorrhage. Twenty-three patients (23/25 [92%]) had intraparenchymal hemorrhage only. One patient (1/25 [4%]) had a combination of intraparenchymal and subarachnoid hemorrhage. One patient (1/25 [4%]) had subdural hemorrhage only. Twenty-two NCCT scans (22/25 [88%]) were performed within 6 hours of presentation. Seventeen NCCT scans (17/25 [68%]) were performed within 3 hours of presentation. CONCLUSION: Despite frequent concerns for intracranial hemorrhage complicating acute stroke and treatment, a low percentage of patients had this complication. Moreover, our frequency is much lower than the wide ranges reported elsewhere. The most common type of intracranial hemorrhage in this cohort was intraparenchymal, but subarachnoid and subdural hemorrhages were also diagnosed and must also be considered. Twenty-eight percent of patients with initial suspicion of acute ischemic stroke are eventually given other diagnoses. These results may have implications for use of CT imaging. PMID- 16219824 TI - Prediction of hemorrhage in acute ischemic stroke using permeability MR imaging. AB - Increased risk of hemorrhagic transformation (HT) limits the general use of thrombolytic therapy in acute ischemic stroke (AIS). This study shows that early blood-brain barrier defects in AIS can be assessed by using permeability MR imaging. Significantly increased permeability was found in 3 patients who later hemorrhaged. This method indicates the potential for identifying patients at higher risk of HT and offers the use of physiologic imaging rather than time from onset of symptoms to guide treatment decisions. PMID- 16219825 TI - Does a relative perfusion measure predict cerebral infarct size? AB - BACKGROUND AND PURPOSE: MR perfusion-weighted imaging (PWI) has been extensively used to quantify cerebral perfusion deficits after the onset of focal ischemia. The present study tested whether a relative measure of cerebral blood flow such as is obtained with PWI is sufficient to predict irreversible tissue damage following focal cerebral ischemia and reperfusion in the rat suture model. METHODS: In rats, the middle cerebral artery was occluded (MCAO) for 1 hour followed by 1-hour reperfusion. Microspheres labeled with different tracers were injected into the left ventricle to permit measurement of blood flow at different time points: before MCAO, 30 minutes post-MCAO and 30 minutes postreperfusion. Absolute cerebral blood flow (CBF) was determined and relative CBF was calculated by comparing absolute CBF at each time point to baseline values before MCAO (relative CBF(B)) or to corresponding contralateral areas in the noninfarcted hemisphere (relative CBF(C)). Infarct size was assessed by 2,3,5 triphenyltetrazolium chloride staining. RESULTS: Absolute CBF in vital tissue was 0.69 +/- 0.07 mL/g/min. In partially and completely necrotic tissue, absolute CBF was 0.39 +/- 0.05 mL/g/min and 0.30 +/- 0.09 mL/g/min, respectively. Although there was a close inverse correlation between infarct volume and absolute CBF (r = 0.79), the correlations between infarct volume and relative CBF(C) were poor (r = 0.21). CONCLUSION: The present study revealed that absolute CBF is superior to relative CBF in predicting irreversible tissue damage following ischemia and reperfusion. PMID- 16219826 TI - Decreased fractional anisotropy of middle cerebellar peduncle in crossed cerebellar diaschisis: diffusion-tensor imaging-positron-emission tomography correlation study. AB - BACKGROUND AND PURPOSE: The purpose of this study was to evaluate the clinical usefulness of diffusion tensor MR imaging (DTI) for the assessment of chronic stroke with crossed cerebellar diaschisis (CCD). METHODS: Twenty-two patients (15 men and 7 women; mean age, 60 years) with chronic stroke (13 ischemic and 9 hemorrhagic) were evaluated by diffusion-tensor MR imaging (DTI) and (18)F fluorodeoxyglucose (FDG)-positron-emission tomography (PET). Fractional anisotropy (FA) and color-coded vector maps were generated. To evaluate afferent fiber systems to the cerebellum, the FA of the bilateral middle cerebellar peduncle (MCP) was measured. Changes of FA values in the MCP were compared against PET results. RESULTS: In patients with a chronic infarct involving more than one-third of the unilateral hemisphere, MCP of the contralesional side showed an FA value of 0.5226 +/- 0.0174, which was significantly lower than that of the ipsilesional side MCP (0.5366 +/- 0.0159) (one-tail paired t test, P = .0009). On FDG-PET scan, decreased glucose metabolism was observed in the affected cerebellum in 19 patients (86.4%). CONCLUSION: DTI can visualize an altered corticocerebellar circuit in the case of chronic stroke with CCD, which is hardly demonstrated by conventional MR images. PMID- 16219827 TI - Advantages and pitfalls in 3T MR brain imaging: a pictorial review. PMID- 16219828 TI - Hyperintense putaminal rim sign is not a hallmark of multiple system atrophy at 3T. AB - BACKGROUND AND PURPOSE: Hyperintense putaminal rim (HPR) on the T2-weighted imaging, which has been observed in our daily practice while reading 3T brain images, has been described as a finding typical of multiple system atrophy (MSA). We hypothesized that the HPR sign is not an exclusive hallmark of MSA at a high magnetic field strength, but rather may be a normal finding. METHODS: Ten consecutive clinically healthy age-matched adults who showed recognizable HPR at 3T were subsequently examined on a 1.5T imaging system within 2 hours. MR examination included axial T2-weighted fast spin-echo (FSE), fluid attenuated inversion recovery (FLAIR) on a 3T scanner, and equivalent T2-weighted FSE at 1.5T. MR images were obtained parallel to the intercommissural plane. All the images were interpreted by 2 experienced neuroradiologists. RESULTS: All 10 subjects (3 men and 7 women; aged 52 +/- 6.1 years [range, 44-61 years], expressed as mean +/- SD) with the positive HPR sign on axial T2-weighted FSE at 3T had negative findings at 1.5T. Such hyperintense rim was also vague or absent on the 3T-FLAIR images. CONCLUSION: Our data suggest that the HPR at 3T scans is a nonspecific, normal finding. FLAIR may be helpful in discriminating between normal subjects and patients with MSA in case of isolated HPR at 3T. PMID- 16219829 TI - Postoperative assessment of extracranial-intracranial bypass by time-resolved 3D contrast-enhanced MR angiography using parallel imaging. AB - PURPOSE: Our goals were to assess image quality of time-resolved contrast enhanced MR angiography (CE MRA), by using 3D data acquisition along with a parallel imaging technique that can improve temporal resolution and to compare this technique with 3D-time-of-flight (TOF) MRA in the postoperative assessment of extracranial (EC)-intracranial (IC) bypass surgery. METHODS: On a 1.5T imaging system, we performed CE MRA by using a 3D fast field-echo sequence in combination with a parallel imaging technique, to obtain images in the coronal plane centered at the postoperative site. Our patient group comprised 17 patients, including 13 after superficial temporal artery-middle cerebral artery (MCA) anastomosis, 3 after external carotid artery-MCA anastomosis, and one after extracranial vertebral artery-posterior cerebral artery anastomosis. Visualization of the anastomosis and the distal flow on the CE-MRA images was assessed comparatively with that on 3D-TOF MR angiograms obtained at the same time. In 6 patients, we also compared the efficiency of visualization on CE-MRA images with that on conventional angiograms. RESULTS: A temporal resolution of 0.8 s/frame could be achieved with the technique employed. The bypass was better demonstrated postoperatively on CE-MRA images than on 3D-TOF MR angiograms in 13 patients (76%), whereas the 2 methods were equivalent in 4 patients (24%). Good correspondence of results was observed in the 6 patients for whom CE MRA and conventional digital subtraction angiography (DSA) images were compared. CONCLUSION: CE MRA by using the parallel imaging technique can increase image acquisition speed with sufficient image quality. This technique is at least equivalent to 3D-TOF MRA to evaluate the postoperative status of EC-IC bypass. PMID- 16219830 TI - The influence of gender on auditory and language cortical activation patterns: preliminary data. AB - PURPOSE AND BACKGROUND: Intersex cortical and functional asymmetry is an ongoing topic of investigation. In this pilot study, we sought to determine the influence of acoustic scanner noise and sex on auditory and language cortical activation patterns of the dominant hemisphere. MATERIALS AND METHODS: Echoplanar functional MR imaging (fMRI; 1.5T) was performed on 12 healthy right-handed subjects (6 men and 6 women). Passive text listening tasks were employed in 2 different background acoustic scanner noise conditions (12 sections/2 seconds TR [6 Hz] and 4 sections/2 seconds TR [2 Hz]), with the first 4 sections in identical locations in the left hemisphere. Cross-correlation analysis was used to construct activation maps in subregions of auditory and language relevant cortex of the dominant (left) hemisphere, and activation areas were calculated by using coefficient thresholds of 0.5, 0.6, and 0.7. RESULTS: Text listening caused robust activation in anatomically defined auditory cortex, and weaker activation in language relevant cortex of all 12 individuals. As a whole, there was no significant difference in regional cortical activation between the 2 background acoustic scanner noise conditions. When sex was considered, men showed a significantly (P < .01) greater change in left hemisphere activation during the high scanner noise rate condition than did women. This effect was significant (P < .05) in the left superior temporal gyrus, the posterior aspect of the left middle temporal gyrus and superior temporal sulcus, and the left inferior frontal gyrus. CONCLUSION: Increase in the rate of background acoustic scanner noise caused increased activation in auditory and language relevant cortex of the dominant hemisphere in men compared with women where no such change in activation was observed. Our preliminary data suggest possible methodologic confounds of fMRI research and calls for larger investigations to substantiate our findings and further characterize sex-based influences on hemispheric activation patterns. PMID- 16219831 TI - Comparison of functional MR imaging guidance to electrical cortical mapping for targeting selective motor cortex areas in neuropathic pain: a study based on intraoperative stereotactic navigation. AB - PURPOSE: To assess the concordance between data from functional MR imaging (fMRI) guidance and the intraoperative electrical cortical mapping (iCM) in targeting selective motor cortex areas in refractory neuropathic pain. METHODS: Twenty-one patients (11 women and 10 men; mean age, 55.6 years) with refractory central (ischemic, 8 cases) and neuropathic pain (trigeminal neuropathy, 6 cases; syrinx/amputation/plexus trauma, 7 cases) underwent surgery for the implantation of an epidural electrode for chronic motor cortex stimulation (MCS) with general anesthesia and a frameless neuronavigation system used for the image-guided targeting procedure. All patients were studied by preoperative fMRI and epidural iCM with somatosensory evoked potentials and motor cortex stimulodetection. fMRI investigated systematically motor tasks of both hands and that related to the somatic area (foot or tongue) affected by pain. fMRI data were analyzed with the Statistical Parametric Mapping99 software (initial analysis threshold [AT] corresponding to P < .001), registered in the neuronavigation system and correlated intraoperatively with iCM. Matching of fMRI and iCM was specifically examined, focusing the study on hand mapping. RESULTS: Concordance between contours of fMRI activation area and iCM in precentral gyrus (mean distance, 3.8 mm) was found in 20/21 patients (95%). Because precision of iCM was suboptimal in 7 patients, concordance for more restrictive values of the AT (P < .0001) was found in only 13 of these 20 patients. Concordance was not found in one patient, as result of image distortion and residual motion artifact. CONCLUSIONS: In this study, fMRI guidance provides information that matches those of an independent functional method. These data illustrate the functional accuracy of fMRI guidance for the operative targeting of selective motor cortex areas in neuropathic pain. PMID- 16219832 TI - Diffusion tensor tractography of the limbic system. AB - BACKGROUND AND PURPOSE: The limbic system, relevant to memory and emotion, is an interesting subject of study in healthy and diseased individuals. It consists of a network of gray matter structures interconnected by white matter fibers. Although gray matter components of this system have been studied by using MR imaging, the connecting fibers have not been analyzed to the same degree. Cerebrospinal fluid (CSF) signal intensity contamination of the fornix and cingulum, the 2 major white matter tracts of the limbic system, can alter diffusion-tensor imaging (DTI) measurements and affect tractography. We investigated the effect of CSF signal intensity suppression on fiber tracking of the limbic connections and characterized the diffusion properties of these structures in healthy volunteers. METHODS: Nine healthy individuals were scanned with standard and CSF-suppressed DTI. Tractography of the fornix and cingulum was performed for both acquisition methods. We report mean diffusivity and fractional anisotropy measurements of the crus, body, and columns of the fornix, and descending, superior, and anterior portions of the cingulum. RESULTS: Diffusion measurements were improved and tractography was facilitated by using CSF suppressed DTI. In particular, tract volume increased, whereas decreases of the mean diffusivity and increases of diffusion anisotropy more accurately represented the underlying tissue by minimizing deleterious partial volume averaging from CSF. This was particularly true for the fornix because it is in closest contact to CSF. Diffusion measurements throughout the limbic connections were consistent in healthy volunteers. CONCLUSION: We recommend the use of CSF suppression when performing diffusion-tensor tractography of the limbic system. PMID- 16219833 TI - Diffusion-tensor MR imaging of the brain in human immunodeficiency virus-positive patients. AB - BACKGROUND AND PURPOSE: There is early evidence that diffusion-tensor imaging (DTI) is useful in demonstrating subtle white matter alterations in different diseases of brain. We hypothesize that DTI in several brain regions in human immunodeficiency virus-positive (HIV+) patients is useful in the early detection of HIV-related brain injury. METHODS: MR imaging and DTI were performed in 60 HIV+ patients and in 30 controls. Fractional anisotropy (FA) and apparent diffusion coefficient (ADC; mm/s(2)) maps were generated and coregistered on T2 weighted images. Regions of interest were placed in the splenium and genu of the callosum, the frontal white matter, and the hippocampus. HIV+ patients were divided into those whose CD4 count were <250 cells/mm(3) or >250 cells/mm(3). According to plasma viral loads, patients were divided into those whose viral loads were <50 copies/mL, 50-100,000 copies/mL, or >100,000 copies/mL. RESULTS: Statistically significant decrease of FA was found in the genu of the corpus callosum in HIV+ patients compared with controls. FA was reduced in the frontal white matter and hippocampi in HIV+ patients compared with controls. Differences, however, were not statistically significant. No statistically significant differences were found between the HIV+ groups for FA of the splenium or between these groups and the controls. ADC values were significantly increased in the genu of HIV+ patients when compared with controls and were also increased in other locations, but without statistical significance. CONCLUSION: As used in this study, DTI was not helpful in identifying patients with early HIV infection. PMID- 16219834 TI - Brain MR imaging abnormalities in kidney transplant recipients. AB - BACKGROUND AND PURPOSE: On the basis of limited available data, brain MR imaging abnormalities in kidney transplant recipients (KTRs) have been predominantly attributed to calcineurin inhibitors (CIs), characteristically presenting as posterior reversible encephalopathy syndrome (PRES). The goal of this study was to evaluate whether CIs play an important role in the incidence, nature, and location of MR imaging brain lesions in adult KTRs by comparing them with dialysis-dependent patients. METHODS: We retrospectively analyzed 98 brain MR imaging examinations in 77 consecutive KTRs presenting with neurologic symptoms from 1990 to 2003. The data were separated into 3 groups according to duration after transplantation of MR imaging: group 1, 0-3 months; group 2, 3-12 months; and group 3, >12 months. Twenty-six MR imaging examinations from 24 additional dialysis-dependent adults were used as controls and comprised group 0. RESULTS: Acute changes (infarcts, infections, PRES) comprised 24% and 19% of lesions in KTRs and group 0 patients, respectively, with infarcts being the most common in all groups. Chronic lesions were responsible for 76% of changes in KTR and 81% in group 0 and were predominantly vascular in etiology. No statistically significant differences in incidence of PRES or other acute changes were found between dialysis-dependent patients and either individual KTR groups or all KTR patients combined. The deep gray matter lesions were more common in KTR, whereas frontal white matter was more frequently affected in patients on dialysis. CONCLUSION: Our study does not support suggestion that MR imaging brain abnormalities in KTR are predominantly due to direct CI toxicity. PMID- 16219835 TI - An exploratory study of ferumoxtran-10 nanoparticles as a blood-brain barrier imaging agent targeting phagocytic cells in CNS inflammatory lesions. AB - BACKGROUND AND PURPOSE: Iron oxide-based contrast agents have been investigated as more specific MR imaging agents for central nervous system (CNS) inflammation. Ferumoxtran-10 is a virus-size nanoparticle, taken up by reactive cells, that allows visualization of the phagocytic components of CNS lesions. Ferumoxtran-10 was compared with standard gadolinium-enhanced MR images in this exploratory trial to assess its potential in evaluation of CNS lesions with inflammatory aspects, including lymphoma, multiple sclerosis (MS), acute disseminated encephalomyelitis (ADEM), and vascular lesions. METHODS: Twenty-three patients with different types of intracranial "inflammatory" lesions underwent standard brain MR with and without gadolinium, followed an average of 10 days later by a ferumoxtran-10 scan. Patients were imaged 24 hours after infusion of 2.6 mg/kg ferumoxtran-10. All MR images were evaluated subjectively by 4 investigators for a difference in enhancement patterns, which could be useful in differential diagnoses. RESULTS: In 5 cases, (one ADEM, 2 stroke, one cavernous venous vascular malformation, one primary central nervous lymphoma) the ferumoxtran-10 scan showed higher signal intensity, larger area of enhancement, or new enhancing areas compared with gadolinium. Most MS patients showed less enhancement with ferumoxtran-10 than with gadolinium. CONCLUSION: Ferumoxtran-10 showed different enhancement patterns in a variety of CNS lesions with inflammatory components in comparison to gadolinium. The impact of timing and therapy need further evaluation to better assess ferumoxtran-10 in addition to gadolinium as contrast agents for use in diagnosis and monitoring therapy in patients with CNS inflammatory lesions. PMID- 16219836 TI - MR Imaging of nonalcoholic Wernicke encephalopathy: a follow-up study. AB - We investigated the correlation of MR imaging features with the pathological evolution and prognosis of nonalcoholic Wernicke encephalopathy. A retrospective review and analysis was conducted of 6 cases of nonalcoholic Wernicke encephalopathy, consisting of MR imaging features, clinical characteristics, and outcomes after thiamine administration. One patient died, 1 patient entered a persistent vegetative state, and the others recovered fully from Wernicke encephalopathy within 2 weeks to 1 year after thiamine administration. Typical MR imaging showed areas of increased T2-weighted and fluid-attenuated inversion recovery (FLAIR) signals symmetrically surrounding the aqueduct and the third ventricle, at the floor of fourth ventricle, in the medial thalami, and in the capita of caudate nuclei. Two patients presenting without coma showed increased T2-weighted and FLAIR signals of the periaqueductal area only. All 4 patients presenting with coma showed increased T2-weighted and FLAIR signals symmetrically in the medial thalami and in the capita of caudate nuclei. Of the 4 patients with coma, 2 patients with deep coma showed increased T2-weighted and FLAIR signals in the medial thalami and caudate nuclei as well as in the frontal and parietal cortices. According to the follow-up results, increased T2-weighted and FLAIR signals in the 4 patients without cortical damage decreased in intensity, consistent with clinical recovery within 2 weeks to 1 year. The patient in a persistent vegetative state exhibited progressive atrophy of the whole brain during the 2 years of the follow-up study. MR imaging is helpful not only to diagnose acute nonalcoholic Wernicke encephalopathy but also to evaluate the pathologic evolution and prognosis of the disorder. PMID- 16219837 TI - MR imaging with diffusion-weighted imaging in acute and chronic Wernicke encephalopathy. AB - Wernicke encephalopathy is a neurologic disorder that results from thiamine deficiency. It is associated with a classic triad of symptoms consisting of ataxia, ocular motor cranial neuropathies, and changes in consciousness. We report 3 cases of Wernicke encephalopathy in which MR imaging, including diffusion-weighted imaging, was performed at the onset and during follow-up. MR imaging findings were correlated with the clinical status of both the acute and chronic stage of Wernicke encephalopathy. PMID- 16219838 TI - Acute encephalopathy associated with intravenous immunoglobulin therapy. AB - Intravenous immunoglobulin therapy is useful against various immune system disorders and viral infections. It is generally safe, and serious adverse reactions are uncommon. We report a rare case of acute encephalopathy following intravenous immunoglobulin therapy for human herpes virus 6 infection in a child. MR imaging findings suggest that the dominant causative mechanism of acute encephalopathy is cytotoxic edema, and the findings indicate 2 primary mechanisms. Reversibility of the restriction of water diffusion (low apparent diffusion coefficient value) on diffusion-weighted MR imaging suggests intramyelinic edema in the myelin sheath, and an increase of glutamate and glutamine complex peak on MR spectroscopy suggests excitotoxic injury to the neurons and astrocytes. PMID- 16219839 TI - Prominent perivenular spaces in multiple sclerosis as a sign of perivascular inflammation in primary demyelination. AB - In this study, we describe prominent perivenular spaces as a sign that is seen on high-resolution (512 x 512) transverse T2-weighted MR images in patients with multiple sclerosis. The observed widening of perivenular space is depicted as a stringlike hyperintensity projecting radially and aligned with multiple sclerosis lesions (usually small), following the course and configuration of deep venular structures. This widening may be an important sign in differentiating primary (ie, in multiple sclerosis) from secondary causes of demyelination. PMID- 16219840 TI - Bilateral cerebral venous angioma associated with varices: a case report and review of the literature. AB - A varix occurring with a cerebral venous angioma is a rare combination of cerebral vascular malformations. The most common vascular anomaly associated with a venous angioma is a cavernous angioma. We report a case of bilateral supratentorial venous angioma with 2 varices involving the angioma situated on the right frontotemporal region. Multiple subependymal veins were seen in the right supratentorial region, which joined to form multiple collector veins, which, in turn, unified into a tortuous channel in the region of the sylvian fissure and continued into venous varices. PMID- 16219841 TI - Angioplasty and stenting in carotid dissection with or without associated pseudoaneurysm. AB - BACKGROUND AND PURPOSE: Carotid angioplasty and stent placement may be the preferred treatment in patients with carotid dissection who have failed medical management. The goal of this study was to determine the procedural feasibility and safety as well as long-term complication rates of carotid angioplasty and stent placement in a consecutive cohort of relatively young, high-surgical-risk patients. PATIENTS AND TECHNIQUES: A series of 26 consecutive patients (mean age, 49 years; 15 men and 11 women) who underwent angioplasty and stent placement for carotid dissection with or without pseudoaneurysm from April 1997 to April 2005 at our institution (9 traumatic, 8 spontaneous, and 9 iatrogenic) was retrospectively reviewed. Twenty-eight stents were used in 29 procedures performed on 27 vessels (20 internal carotid arteries and 7 common carotid arteries). Patients were followed with cerebral angiography, CT, sonography, or clinically for a mean of 14.6 months (range, 5 days to 48.2 months) with 17 of 26 patients having at least 6 months of follow-up. Procedural and long-term complication rates were calculated. RESULTS: Dissection-induced stenosis was reduced from 71 +/- 18% to no significant stenosis in 20 of the 21 patients with measurable stenosis. The procedural transient ischemic attack (TIA) rate was 3 of 29 procedures (10.3%). There were no procedural strokes. One patient required angioplasty of a common femoral artery. One procedure was terminated when an asymptomatic new intimal flap was created before intervention. Two patients had occlusions of the treated vessel noted at 22 days (presented with contralateral stroke) and 3.4 months (asymptomatic). There were 2 unrelated deaths from myocardial infarction at 8 days and 15.2 months. Two patients had recurrent ipsilateral TIA at 2.7 months and 12 months. The 30-day occlusion and death rate was 2 of 29 procedures (6.9%). CONCLUSION: In this series, angioplasty and stent placement were effective in relieving stenosis secondary to carotid dissection with or without pseudoaneurysm and have low rates of ischemic complications. PMID- 16219842 TI - MR and clinical follow-up of diffusion-weighted cerebral lesions after carotid artery stenting. AB - PURPOSE: The purpose of this prospective study was to determine the outcome of postprocedural cerebral diffusion-weighted (DW) MR lesions after carotid artery stent placement (CAS) and the incidence of new cerebral MR lesions 6 months after the procedure. MATERIALS AND METHODS: DW and T2-weighted MR imaging of the brain and neurologic examinations were performed in 105 patients before and 24 hours and 6 months after CAS. In addition, a selective control angiography of the treated artery was performed after 6 months. RESULTS: In 22 (21%) of 105 patients, DW MR images 24 hours after CAS showed 64 new neurologically silent lesions; 2 (3.1%) of these 64 lesions were also visible in T2-weighted MR images. The latter ones were still visible after 6 months. In the remaining 62 lesions, there were no abnormalities visible in DW and T2-weighted imaging at follow-up. In 2 (1.9%) of the 105 patients, new cerebral lesions were seen in T2-weighted images after 6 months; one patient was neurologically symptomatic. All others patients were neurologically unremarkable at 6-month follow-up. CONCLUSIONS: Most postprocedural DW lesions showed no manifestations at 6-month MR follow-up and were clinically silent. This indicates that these lesions are potentially reversible and of no major neurologic sequelae. In addition, follow-up 6 months after CAS showed a very low incidence of new cerebral lesions and neurologic events. PMID- 16219843 TI - The use of balloon-expandable stents in the management of intracranial arterial diseases: a 5-year single-center experience. AB - BACKGROUND AND PURPOSE: Although stent-assisted intracranial procedures are becoming a routine clinical practice, there have been relatively few large studies published in the literature regarding the application of the balloon expandable stent technology in the treatment of intracranial arterial diseases. In this report, the authors reviewed their experience with 75 cases at a single center. METHODS: From 1998 to 2003, 75 patients underwent percutaneous transluminal intracranial stent placement as a treatment for wide-necked intracranial aneurysms and atherosclerotic stenoses. The anatomy of the target lesions, technical details of the procedures, device functionality, procedure related complications, and short-term outcomes were reviewed in a retrospective fashion. RESULTS: The clinical indications included wide-necked intracranial aneurysms (59) and atherosclerotic stenoses (16). The stent was successfully deployed in 92% of the patients (69 of 75 cases). In the remaining 6 cases, the causes of failed stent deployment included arterial tortuosities (2), stent migration (2), fracture of the stent (1), and arterial perforation (1). The short term outcome (mean follow-up, 7.5 months; range, 3-12 months) was evaluated by using the modified Rankin scale (MR spectroscopy 0-6). Fifty-three patients (70.6%) had excellent outcomes (MR spectroscopy 0-1), 12 (16%) had good outcomes (MR spectroscopy 2), and 5 (6.7%) had poor outcomes (MR spectroscopy 4-5). Five patients (6.7%) died. CONCLUSION: The use of BES is associated with a high rate of hemorrhagic and ischemic complications, more specifically when used in the anterior circulation. Cases of large-necked aneurysms not treatable with balloon remodeling technique and atheromatous sclerosis could be eligible for this treatment. PMID- 16219844 TI - CT angiography and MR angiography in the evaluation of carotid cavernous sinus fistula prior to embolization: a comparison of techniques. AB - BACKGROUND AND PURPOSE: This study compared CT angiography (CTA), MR angiography (MRA), and digital subtraction angiography (DSA) in elucidating the size and location of carotid cavernous sinus fistulas (CCFs) before embolization treatment. METHODS: This was a retrospective study of 53 patients with angiographically confirmed CCF. All patients underwent pre- and postcontrast enhanced CTA and DSA, and 50 patients also underwent MRA. Two neuroradiologists rated detectability of the fistula tract as "good," "moderate," or "poor" in source images obtained by using each procedure. The chi(2) test was used to compare the imaging modalities with respect to their ability to detect fistulas. RESULTS: CTA did not differ significantly from DSA (P = .155), and both CTA (P = .001) and DSA (P = .007) performed significantly better than MRA in the population as a whole. Differences in performance among the methods, however, depended upon the segmental location of the fistula along the internal carotid artery (ICA). CTA and MRA were similar in detection of CCFs in patients with a fistula at segment 3. CTA significantly outperformed MRA in patients with a fistula at segment 4, who accounted for approximately half of the population. CONCLUSIONS: CTA source imaging has proved itself as useful as DSA for detecting CCFs. Of the 2 noninvasive techniques, CTA performed better than MRA in the population as a whole and in most patients whose fistula was located at segment 4 or 5 of the ICA. PMID- 16219845 TI - Correlation between lumenal geometry changes and hemodynamics in fusiform intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Hemodynamics may predispose aneurysms to rupture; however, hemodynamic descriptors that can describe aneurysm growth are not well understood. We examined the relationship between hemodynamics and growth of 2 fusiform basilar artery aneurysms in an effort to define hemodynamic variables that may be helpful in predicting aneurysmal growth. METHODS: Two patients with basilar fusiform aneurysms of a similar size were followed for a 2-year period. The lumenal geometry and inflow and outflow rates were acquired by using MR angiography and velocimetry, respectively. The location of aneurysmal growth was identified by coregistering aneurysm models that were acquired at different times. Hemodynamic descriptors were calculated by using computational fluid dynamic simulations and compared with aneurysm growth pattern. RESULTS: One patient had an aneurysm that grew significantly, but a similar-sized aneurysm in the other patient remained unchanged. The largest aneurysmal growth (approximately 3 mm/year) was found at the inferior side (lower part) of the aneurysm, where the wall shear stress was very low (<0.1N/m(2)). The general flow patterns did not change with time, even in the aneurysm that grew, but histograms of wall shear stress were very different in these 2 patients. CONCLUSIONS: This study investigates whether hemodynamic descriptors can be correlated with regional changes in aneurysm lumen morphology on a patient-specific basis. That capability will permit the testing, in longitudinal studies, of hypotheses as to which mechanisms are the most important in aneurysm growth. PMID- 16219846 TI - Can neck size in elastase-induced aneurysms be controlled? A prospective study. AB - BACKGROUND AND PURPOSE: An earlier retrospective study indicated that the neck size of elastase-induced aneurysms could be controlled by adjusting the position of the inflated balloon. We report the current prospective study to confirm our previous work. METHODS: Ninety elastase-induced aneurysms were created in rabbits. Group 1 (n = 62) included cases in which the occlusion balloon resided low, completely within the brachiocephalic/subclavian arteries. Group 2 (n = 28) included cases in which the balloon resided high, within both the common carotid artery and brachiocephalic/subclavian arteries. Follow-up digital subtraction angiography was performed. The aneurysm sizes were measured and compared between groups. The Student t test and the Fisher exact test were used for statistical analysis. RESULTS: The mean aneurysm neck diameter and width for group 1 was significantly larger than that of group 2 (3.4 +/- 1.2 and 2.3 +/- 0.9 mm, P < .001; 3.8 +/- 1.0 and 3.3 +/- 0.9 mm, P < .05, respectively). The proportion of wide-necked aneurysms in group 1 was significantly larger than that in group 2 (29% vs 4%; P < .005). Mean dome-to-neck ratios were 1.2 +/- 0.4 and 1.7 +/- 0.7 for groups 1 and 2 (P < .005). There was no significant difference in aneurysm height between groups 1 and 2 (8.0 +/- 1.7 and 7.5 +/- 2.2 mm; P > .05). CONCLUSION: The neck size of elastase-induced aneurysm models in rabbits can be controlled by adjusting the position of the inflated balloon. PMID- 16219847 TI - Stent-assisted coiling of intracranial aneurysms aided by virtual parent artery reconstruction. AB - The availability of stents designed specifically for use in the intracranial vasculature has increased the use of stent-assisted coiling for treatment of wide necked and complex intracranial aneurysms. We present a technique for pretreatment planning and visualization of a virtual stent within the parent artery by using a virtual reconstruction of the parent artery across the aneurysm neck. As illustrated by 2 clinical examples, this method provides information not otherwise available regarding the location of portions of the stent that are not visible on fluoroscopy. During treatment, this information enhances the ability to determine the location of coils in relation to the stent boundaries and should thereby improve the ability to avoid parent artery compromise. PMID- 16219849 TI - Proatlantal intersegmental arteries of external carotid artery origin associated with Galen's vein malformation. AB - Carotid basilar anastomoses can occasionally persist beyond the embryonic period. These anomalies are most often incidentally detected in adulthood, during workups for unrelated pathologies. Persistence of the proatlantal intersegmental arteries is a rare form of primitive carotid-basilar anastomoses. Bilateral proatlantal inter- segmental arteries are an extremely rare occurrence, of which only 3 cases have been reported in the literature. An analysis of vascular anomalies associated with Galen's vein malformations revealed 3 children in whom persistence of type II proatlantal arteries was seen. These included one child in whom proatlantal arteries were persistent bilaterally. We report the clinical and angiographic findings and discuss the embryologic and therapeutic implications of this unique association. PMID- 16219850 TI - Volumetric MR imaging of oral, maxillary sinus, oropharyngeal, and hypopharyngeal cancers: correlation between tumor volume and lymph node metastasis. AB - BACKGROUND AND PURPOSE: The size of the primary lesion and the lymph node metastasis are critical indicators for the patient prognosis. Here we attempted to assess the correlation between these 2 prognostic parameters in patients with oropharyngeal, hypopharyngeal, oral, or maxillary sinus cancer. METHODS: We retrospectively studied 66 patients with oropharyngeal or hypopharyngeal (n = 24), oral (n = 35), or maxillary sinus (n = 7) cancer. Of these patients, 25 (10 with oral or maxillary sinus and 15 with pharyngeal cancers) had lymph node metastases. We measured the volumes of the primary lesions as sums of gadolinium enhanced areas on fat-suppressed, spectral presaturation with inversion recovery (SPIR) T1-weighted images. Histologically confirmed metastatic nodes were mapped to the neck levels. RESULTS: The tumor volumes were well correlated with the clinical T-category for the primary lesions in oral, maxillary sinus, and oropharyngeal and hypopharyngeal cancers. The volumes of the oropharyngeal and hypopharyngeal cancers were significantly greater (P < .05) in patients with metastatic nodes than in patients without metastasis, whereas there was no significant correlation between the tumor volume and nodal metastasis in patients with oral or maxillary sinus cancer (Mann-Whitney U test). Furthermore, the correlation between tumor volume and the distribution of metastatic nodes in the neck was observed to be weak in patients with oral or maxillary sinus cancer. CONCLUSION: The MR image-based tumor volume measurement proved to be clinically feasible. We observed a good correlation between tumor volume and lymph node metastasis in patients with oropharyngeal or hypopharyngeal cancer but not in those with oral or maxillary sinus cancer. PMID- 16219848 TI - A quantitative MR imaging assessment of leukoencephalopathy in children treated for acute lymphoblastic leukemia without irradiation. AB - PURPOSE: Intravenous methotrexate (IV-MTX), an effective treatment for acute lymphoblastic leukemia (ALL), has a significant toxic effect on the central nervous system, with leukoencephalopathy (LE) being the most common form. The purpose of this study was to use objective quantitative MR imaging to prospectively assess the temporal evolution of LE extent and intensity. METHODS: Forty-five children (low-risk, 10 mol/L/12F; mean age, 5.0 years at diagnosis; standard/high-risk, 11 mol/L/12F; mean age, 9.2 years at diagnosis) treated for ALL on a single institutional protocol were evaluated longitudinally to assess the extent of LE (proportion of white matter impacted) through tissue segmentation and the relative intensity of LE through relative elevations in T1 and T2 relaxation rates. One-sided Wilcoxon-Mann-Whitney tests were used to assess differences in quantitative measures at 4 different points in therapy both within and between risk arms. RESULTS: The proportion of white matter affected in both patient groups increased significantly with additional courses of IV-MTX, whereas the intensity of LE also increased steadily; however, both the intensity and extent of LE declined significantly approximately 1.5 years after completion of IV-MTX. Increases in the T1 and T2 relaxation rates above normal-appearing white matter were significantly correlated with each other and were dependent on the proportion of white matter affected. CONCLUSION: Higher doses and more courses of IV-MTX were associated with increased intensity and extent of LE. There was a significant reduction in both the intensity and extent of LE after completion of therapy. The impact of these changes on neurocognitive functioning and quality of life in survivors remains to be determined. PMID- 16219851 TI - Silent sinus syndrome: an acquired condition. AB - We present the case of a 30-year-old man with silent sinus syndrome. A CT scan obtained 10 years earlier showed completely normal maxillary sinuses. This case illustrates the acquired nature of this disorder. PMID- 16219852 TI - CT-demonstrated transcalvarial channels diagnostic of dural arteriovenous fistula. AB - MR imaging-evident intraosseous channels associated with the presence of a dural arteriovenous fistula have been described in the literature. We describe these channels in relation to a case of dural arteriovenous fistula seen on CT. The presence of this subtle sign should be sought, and the area reviewed in all CT evaluations of patients with pulsatile tinnitus. PMID- 16219853 TI - Intradiskal extravasation with low-volume cement filling in percutaneous vertebroplasty. AB - BACKGROUND AND PURPOSE: Adjacent fracture formation after percutaneous vertebroplasty has been reported in literature. The purpose of this study was to determine whether intervertebral disk extravasation is related to adjacent fracture formation in low-volume cement-filling vertebroplasty. METHODS: A retrospective analysis of 308 patients having vertebroplasty was undertaken. Anteroposterior and lateral procedural fluoroscopy radiographs were analyzed for mild, moderate, or severe disk extravasation. Symptomatic refracture location relative to the presence or absence of extravasation was analyzed on follow-up fluoroscopy radiographs. RESULTS: Of the 308 patients in the study, 81 patients had disk extravasation at a total of 85 vertebral levels; 40 levels had mild extravasation, 38 levels had moderate extravasation, and 7 had severe extravasation. Of the 40 levels with mild extravasation, 6 new adjacent fractures occurred next to the disk leakage. Of the 38 with moderate extravasation, 6 new adjacent fractures occurred. Of the 7 levels of severe extravasation, only one new adjacent fracture occurred. Hence, a total of 13 adjacent fractures occurred next to disk extravasation; 13 nonadjacent fractures also occurred in the patients with disk extravasation. In the patients without disk extravasation, 28 adjacent and 24 nonadjacent subsequent fractures occurred. The average available cement volumes injected into vertebral bodies causing disk leakage ranged from 4.78 to 5.60 mL. CONCLUSIONS: With low-volume cement-filling percutaneous vertebroplasty, we cannot conclude (level of significance alpha = .05) that for patients who have a new fracture there is significance between the location of the fracture and the occurrence of disk extravasation. PMID- 16219854 TI - Spinal epidural synovial sarcoma: a case of homogeneous enhancing large paravertebral mass on MR imaging. AB - We report the MR imaging findings in a 44-year-old man with a low-grade synovial sarcoma. There was a right-sided epidural and paravertebral mass, widening of the ipsilateral neural foramen at the L4-L5 level, and focal erosion of the right superior articular process of the L5 vertebra. The mass was relatively homogeneous, hyperintense to muscle and isointense to fat on T2-weighted images, and isointense to muscle on T1-weighted images, and it demonstrated moderate homogeneous enhancement. PMID- 16219855 TI - Nonenhancing spinal subdural metastatic tumor. AB - We describe a case of a spinal subdural metastatic tumor that became rapidly symptomatic after a minor trauma, as a result of severe cord compression and cord hemorrhage. On MR imaging, the lesion was oval, hyperintense with a dark rim on T2-weighted fast spin-echo images, isointense to the cord on T1-weighted images, and had dark and bright areas on gradient-echo T2*-weighted images, consistent with a hyperacute-to-acute hematoma. The hemorrhagic tumor showed no evidence of contrast enhancement. PMID- 16219856 TI - Oligodendroglioma metastatic to bone marrow. AB - We report on a patient with oligodendroglioma metastatic to bone, presenting with pancytopenia and fever 10 years after initial tumor resection. Our review of the literature showed a total of 30 reported extraneural metastases, with only 19 of these being similar cases of bone metastases. These bony lesions have increased signal intensity in T2-weighted and low signal intensity on T1-weighted images, with intense homogeneous enhancement. However, on MR imaging, we were unable to find necrosis or compression deformity of the vertebrae, despite extensive metastatic disease. PMID- 16219857 TI - Subarachnoid hemorrhage due to isolated spinal artery aneurysm in four patients. AB - Spinal artery aneurysms are usually found with arteriovenous malformations or other entities that increase hemodynamic stress. Isolated spinal artery aneurysms are rare. Four patients who presented with the acute onset of lower back pain underwent MR imaging, which revealed spinal subarachnoid hemorrhage. In all patients, work-up yielded a diagnosis of isolated spinal aneurysm, and operative treatment was successful. In the appropriate clinical setting, spinal aneurysm should be considered as a possible cause of spinal subarachnoid hemorrhage. PMID- 16219864 TI - Reporting terminology for lumbar disk herniations: axial segmentation of the preneural foraminal portion of the lumbar nerve roots. PMID- 16219858 TI - Visualization of sacral nerve roots via percutaneous intraspinal navigation (PIN). AB - A percutaneous technique for visualizing sacral nerve roots is described. A fiberscope was inserted into the subarachnoid space through a sheath that was inserted via a percutaneous lumbar puncture. The sacral nerve roots were identified with endoscopic visualization and x-ray fluoroscopy localization of the endoscope. These images were compared with those obtained from a videoscope, which revealed better imaging. Specific sacral nerve roots can be identified by using a combination of endoscopy and x-ray fluoroscopy. This technique may enable minimally invasive interventions such as lysis of adhesions, arachnoid cyst decompression, selective dorsal rhizotomy, and more selective and precise nerve stimulation electrode placement. PMID- 16219865 TI - Location, location, location: angiography discerns early MR imaging vessel signs due to proximal arterial occlusion and distal collateral flow. PMID- 16219866 TI - Are cervical nerve root blocks "safe and effective"? PMID- 16219867 TI - Re: angiographic evidence of aneurysm neck healing following endovascular treatment with bioactive coils. PMID- 16219868 TI - Evidence for Nr4a1 as a cold-induced effector of brown fat thermogenesis. AB - Acute cold exposure leads to norepinephrine release in brown adipose tissue (BAT) and activates uncoupling protein (UCP)1-mediated nonshivering thermogenesis. Chronic sympathetic stimulation is known to initiate mitochondrial biogenesis, UCP1 expression, hyperplasia of BAT, and recruitment of brown adipocytes in white adipose tissue (WAT) depots. Despite distinct functions of BAT and WAT in energy balance, only a few genes are exclusively expressed in either tissue. We identified NUR77 (Nr4a1), an orphan receptor, to be induced transiently in brown adipocytes in response to beta-adrenergic stimulation and in BAT of cold-exposed mice. Subsequent reporter gene assays demonstrated an inhibitory action of NUR77 on basal and peroxisome proliferator-activated receptor (PPAR)gamma/retinoid X receptor (RXR)alpha-mediated transactivation of the Ucp1 enhancer in heterologous cotransfection experiments. Despite this function of NUR77 in the control of Ucp1 gene expression, nonshivering thermogenesis was not affected in Nur77 knockout mice. However, we observed a superinduction of Nor1 in BAT of cold-exposed knockout mice. We conclude that NUR77 is a cold-induced negative regulator of Ucp1, but phenotypic consequences in knockout mice are compensated by functional redundancy of Nor1. PMID- 16219869 TI - Chronic heart rate reduction remodels ion channel transcripts in the mouse sinoatrial node but not in the ventricle. AB - We investigated the effects of chronic and moderate heart rate (HR) reduction on ion channel expression in the mouse sinoatrial node (SAN) and ventricle. Ten-week old male C57BL/6 mice were treated twice daily with either vehicle or ivabradine at 5 mg/kg given orally during 3 wk. The effects of HR reduction on cardiac electrical activity were investigated in anesthetized mice with serial ECGs and in freely moving mice with telemetric recordings. With the use of high-throughput real-time RT-PCR, the expression of 68 ion channel subunits was evaluated in the SAN and ventricle at the end of the treatment period. In conscious mice, ivabradine induced a mean 16% HR reduction over a 24-h period that was sustained over the 3-wk administration. Other ECG parameters were not modified. Two-way hierarchical clustering analysis of gene expression revealed a separation of ventricles from SANs but no discrimination between treated and untreated ventricles, indicating that HR reduction per se induced limited remodeling in this tissue. In contrast, SAN samples clustered in two groups depending on the treatment. In the SAN from ivabradine-treated mice, the expression of nine ion channel subunits, including Navbeta1 (-25%), Cav3.1 (-29%), Kir6.1 (-28%), Kvbeta2 (-41%), and Kvbeta3 (-30%), was significantly decreased. Eight genes were significantly upregulated, including K+ channel alpha-subunits (Kv1.1, +30%; Kir2.1, +29%; Kir3.1, +41%), hyperpolarization-activated cation channels (HCN2, +24%; HCN4, +52%), and connexin 43 (+26%). We conclude that reducing HR induces a complex remodeling of ion channel expression in the SAN but has little impact on ion channel transcripts in the ventricle. PMID- 16219870 TI - The cover. Self-portrait after the Spanish influenza. PMID- 16219871 TI - Genes help unravel Marfan pathology, point way to potential new therapies. PMID- 16219872 TI - NIH eases ethics rules on employees: consulting ban to remain. PMID- 16219873 TI - Pain studies illuminate the placebo effect. PMID- 16219874 TI - To Err Is Human 5 years later. PMID- 16219875 TI - To Err Is Human 5 years later. PMID- 16219876 TI - Access to trauma centers. PMID- 16219877 TI - Abciximab in the treatment of ST-segment elevation myocardial infarction. PMID- 16219878 TI - Climbing a triassic Mount Everest: into thinner air. PMID- 16219879 TI - Comorbidity and survival disparities among black and white patients with breast cancer. AB - CONTEXT: Reasons for the shorter survival of black breast cancer patients compared with their white counterparts are not completely understood. OBJECTIVE: To evaluate the role of comorbidity in this racial disparity among breast cancer patients. DESIGN, SETTING, AND PATIENTS: Historical cohort from the Henry Ford Health System (a large comprehensive health system in Detroit, Mich) followed up for a median of 10 years. Patients (n = 906) included 264 black (29.1%) and 642 white (70.9%) women diagnosed as having breast cancer between 1985 and 1990. Detailed comorbidity data (268 comorbidities) and study data were abstracted from medical records and institutional, Surveillance, Epidemiology, and End Results, and Michigan State registries. Associations were analyzed with logistic and Cox regression. MAIN OUTCOME MEASURES: Breast cancer recurrence/progression and survival to death from all, breast cancer, and competing (non-breast cancer) causes. RESULTS: Of blacks, 64 (24.9%) died of breast cancer and 95 (37.0%) died of competing causes. Comparable data for whites were 115 (18.3%) and 202 (32.1%). Blacks had worse all-cause survival (hazard ratio [HR], 1.34; 95% confidence interval [CI], 1.11-1.62), breast cancer-specific survival (HR, 1.47; 95% CI, 1.08-2.00), and competing-causes survival (HR, 1.27; 95% CI, 1.00-1.63). A total of 77 adverse comorbidities were associated with reduced survival. Adverse comorbidity count was associated with all-cause (adjusted HR, 1.29; 95% CI, 1.19 1.40) and competing-causes survival but was not associated with recurrence/progression or breast cancer-specific survival. At least 1 adverse comorbidity was observed in 221 (86.0%) blacks and 407 (65.7%) whites (odds ratio, 3.20; 95% CI, 2.17-4.72). Comparisons of unadjusted and comorbidity adjusted HRs indicated that adverse comorbidity explained 49.1% of all-cause and 76.7% of competing-causes survival disparity. Diabetes and hypertension were particularly important in explaining disparity. CONCLUSIONS: More black breast cancer patients die of competing causes than of breast cancer. Effective control of comorbidity in black breast cancer patients should help improve life expectancy and lead to a reduction in survival disparities. PMID- 16219880 TI - Trends in serum lipids and lipoproteins of adults, 1960-2002. AB - CONTEXT: Serum total and low-density lipoprotein (LDL) cholesterol contribute significantly to atherosclerosis and its clinical sequelae. Previous analyses of data from the National Health and Nutrition Examination Surveys (NHANES) showed that mean levels of total cholesterol of US adults had declined from 1960-1962 to 1988-1994, and mean levels of LDL cholesterol (available beginning in 1976) had declined between 1976-1980 and 1988-1994. OBJECTIVE: To examine trends in serum lipid levels among US adults between 1960 and 2002, with a particular focus on changes since the 1988-1994 NHANES survey. DESIGN, SETTING, AND PARTICIPANTS: Blood lipid measurements taken from 6098 to 15 719 adults who were examined in 5 distinct cross-sectional surveys of the US population during 1960-1962, 1971 1974, 1976-1980, 1988-1994, and 1999-2002. MAIN OUTCOME MEASURES: Mean serum total cholesterol, LDL cholesterol, high-density lipoprotein (HDL) cholesterol, and geometric mean serum triglyceride levels, and the percentage of adults with a serum total cholesterol level of at least 240 mg/dL (> or =6.22 mmol/L). RESULTS: Between 1988-1994 and 1999-2002, total serum cholesterol level of adults aged 20 years or older decreased from 206 mg/dL (5.34 mmol/L) to 203 mg/dL (5.26 mmol/L) (P=.009) and LDL cholesterol levels decreased from 129 mg/dL (3.34 mmol/L) to 123 mg/dL (3.19 mmol/L) (P<.001). Greater and significant decreases were observed in men 60 years or older and in women 50 years or older. The percentage of adults with a total cholesterol level of at least 240 mg/dL (> or =6.22 mmol/L) decreased from 20% during 1988-1994 to 17% during 1999-2002 (P<.001). There was no change in mean HDL cholesterol levels and a nonsignificant increase in geometric mean serum triglyceride levels (P = .06). CONCLUSIONS: The decrease in total cholesterol level observed during 1960-1994 and LDL cholesterol level observed during 1976-1994 has continued during 1999-2002 in men 60 to 74 years and women 50 to 74 years. The target value of no more than 17% of US adults with a total cholesterol level of at least 240 mg/dL (> or =6.22 mmol/L), an objective of Healthy People 2010, has been attained. The increase in the proportion of adults using lipid-lowering medication, particularly in older age groups, likely contributed to the decreases in total and LDL cholesterol levels observed. The increased prevalence of obesity in the US population may have contributed to the increase in mean serum triglyceride levels. PMID- 16219881 TI - Incidence of end-stage renal disease in patients with type 1 diabetes. AB - CONTEXT: End-stage renal disease (ESRD) is one of the most severe complications of type 1 diabetes. Yet, data on patients' risk of developing ESRD are sparse. OBJECTIVES: To estimate the long-term risk of developing ESRD and to assess how age at diagnosis of diabetes, time period of diagnosis, and sex affect the risk. DESIGN, SETTING, AND PATIENTS: A cohort of all patients younger than 30 years diagnosed as having type 1 diabetes in Finland in 1965-1999 (n = 20,005) was identified from the Finnish Diabetes Register. The cohort was followed up from diagnosis of diabetes until development of ESRD (dialysis or kidney transplantation as identified from the Finnish Registry for Kidney Diseases), death, or end of follow-up on December 31, 2001. MAIN OUTCOME MEASURE: Cumulative incidence of ESRD, accounting for death as a competing risk. RESULTS: The cohort was followed up for maximally 37 years, with a median of 16.7 years. During 346 851 person-years, 632 patients developed ESRD. The cumulative incidence of ESRD was 2.2% at 20 years and 7.8% at 30 years after diagnosis. The risk of developing ESRD was lowest in patients whose diagnosis occurred at younger than 5 years. The risk of ESRD was lower for patients diagnosed as having type 1 diabetes in later years. The risk did not differ significantly between sexes. CONCLUSIONS: With regard to ESRD, the prognosis of type 1 diabetes has improved during the past 4 decades. Children diagnosed as having diabetes before age 5 years have the most favorable prognosis. Overall, incidence of ESRD appears to be lower than previously estimated. PMID- 16219882 TI - Early experience with pay-for-performance: from concept to practice. AB - CONTEXT: The adoption of pay-for-performance mechanisms for quality improvement is growing rapidly. Although there is intense interest in and optimism about pay for-performance programs, there is little published research on pay-for performance in health care. OBJECTIVE: To evaluate the impact of a prototypical physician pay-for-performance program on quality of care. DESIGN, SETTING, AND PARTICIPANTS: We evaluated a natural experiment with pay-for-performance using administrative reports of physician group quality from a large health plan for an intervention group (California physician groups) and a contemporaneous comparison group (Pacific Northwest physician groups). Quality improvement reports were included from October 2001 through April 2004 issued to approximately 300 large physician organizations. MAIN OUTCOME MEASURES: Three process measures of clinical quality: cervical cancer screening, mammography, and hemoglobin A1c testing. RESULTS: Improvements in clinical quality scores were as follows: for cervical cancer screening, 5.3% for California vs 1.7% for Pacific Northwest; for mammography, 1.9% vs 0.2%; and for hemoglobin A1c, 2.1% vs 2.1%. Compared with physician groups in the Pacific Northwest, the California network demonstrated greater quality improvement after the pay-for-performance intervention only in cervical cancer screening (a 3.6% difference in improvement [P = .02]). In total, the plan awarded 3.4 million dollars (27% of the amount set aside) in bonus payments between July 2003 and April 2004, the first year of the program. For all 3 measures, physician groups with baseline performance at or above the performance threshold for receipt of a bonus improved the least but garnered the largest share of the bonus payments. CONCLUSION: Paying clinicians to reach a common, fixed performance target may produce little gain in quality for the money spent and will largely reward those with higher performance at baseline. PMID- 16219883 TI - Impact of candesartan on nonfatal myocardial infarction and cardiovascular death in patients with heart failure. AB - CONTEXT: Angiotensin-converting enzyme (ACE) inhibitors reduce the risk of myocardial infarction (MI), but it is not known whether angiotensin receptor blockers have the same effect. OBJECTIVE: To assess the impact of the angiotensin receptor blocker candesartan on MI and other coronary events in patients with heart failure. DESIGN, SETTING, AND PARTICIPANTS: The Candesartan in Heart Failure: Assessment of Reduction in Mortality and Morbidity (CHARM) program, a randomized, placebo-controlled study enrolling patients (mean age, 66 [SD, 11] years) with New York Heart Association class II to IV symptoms who were randomly allocated to receive candesartan (target dose, 32 mg once daily) or matching placebo given in addition to optimal therapy for heart failure. Patients were enrolled from March 1999 through March 2001. Of 7599 patients allocated, 4004 (53%) had experienced a previous MI, and 1808 (24%) currently had angina. At baseline, 3125 (41%) were receiving an ACE inhibitor; 4203 (55%), a beta-blocker; 3153 (42%), a lipid-lowering drug; 4246 (56%), aspirin; and 6286 (83%), a diuretic. MAIN OUTCOME MEASURE: The primary outcome of the present analysis was the composite of cardiovascular death or nonfatal MI in patients with heart failure receiving candesartan or placebo. RESULTS: During the median follow-up of 37.7 months, the primary outcome of cardiovascular death or nonfatal MI was significantly reduced in the candesartan group (775 patients [20.4%]) vs the placebo group (868 [22.9%]) (hazard ratio [HR], 0.87; 95% confidence interval [CI], 0.79-0.96; P = .004; number needed to treat [NNT], 40). Nonfatal MI alone was also significantly reduced in the candesartan group (116 [3.1%]) vs the placebo group (148 [3.9%]) (HR, 0.77; 95% CI, 0.60-0.98; P = .03; NNT, 118). The secondary outcome of fatal MI, sudden death, or nonfatal MI was significantly reduced with candesartan (459 [12.1%]) vs placebo (522 [13.8%]) (HR, 0.86; 95% CI, 0.75-0.97; P = .02; NNT, 59). Risk reductions in cardiovascular death or nonfatal MI were similar across predetermined subgroups and the component CHARM trials. There was no impact on hospitalizations for unstable angina or coronary revascularization procedures with candesartan. CONCLUSION: In patients with heart failure, candesartan significantly reduces the risk of the composite outcome of cardiovascular death or nonfatal MI. PMID- 16219885 TI - Palliative sedation in dying patients: "we turn to it when everything else hasn't worked". AB - Despite skilled palliative care, some dying patients experience distressing symptoms that cannot be adequately relieved. A patient with metastatic breast cancer, receiving high doses of opioids administered to relieve pain, developed myoclonus. After other approaches proved ineffective, palliative sedation was an option of last resort. The doctrine of double effect, the traditional justification for palliative sedation, permits physicians to provide high doses of opioids and sedatives to relieve suffering, provided that the intention is not to cause the patient's death and that certain other conditions are met. Such high doses are permissible even if the risk of hastening death is foreseen. Because intention plays a key role in this doctrine, clinicians must understand and document which actions are consistent with an intention to relieve symptoms rather than to hasten death. The patient or family should agree with plans for palliative sedation. The attending physician needs to explain to them, as well as to the medical and nursing staff, the details of care and the justification for palliative sedation. Because cases involving palliative sedation are emotionally stressful, the patient, family, and health care workers can all benefit from talking about the complex medical, ethical, and emotional issues they raise. PMID- 16219886 TI - Legal concerns and the influenza vaccine shortage. PMID- 16219887 TI - Pay-for-performance research: how to learn what clinicians and policy makers need to know. PMID- 16219884 TI - Plasma fibrinogen level and the risk of major cardiovascular diseases and nonvascular mortality: an individual participant meta-analysis. AB - CONTEXT: Plasma fibrinogen levels may be associated with the risk of coronary heart disease (CHD) and stroke. OBJECTIVE: To assess the relationships of fibrinogen levels with risk of major vascular and with risk of nonvascular outcomes based on individual participant data. DATA SOURCES: Relevant studies were identified by computer-assisted searches, hand searches of reference lists, and personal communication with relevant investigators. STUDY SELECTION: All identified prospective studies were included with information available on baseline fibrinogen levels and details of subsequent major vascular morbidity and/or cause-specific mortality during at least 1 year of follow-up. Studies were excluded if they recruited participants on the basis of having had a previous history of cardiovascular disease; participants with known preexisting CHD or stroke were excluded. DATA EXTRACTION: Individual records were provided on each of 154,211 participants in 31 prospective studies. During 1.38 million person years of follow-up, there were 6944 first nonfatal myocardial infarctions or stroke events and 13,210 deaths. Cause-specific mortality was generally available. Analyses involved proportional hazards modeling with adjustment for confounding by known cardiovascular risk factors and for regression dilution bias. DATA SYNTHESIS: Within each age group considered (40-59, 60-69, and > or =70 years), there was an approximately log-linear association with usual fibrinogen level for the risk of any CHD, any stroke, other vascular (eg, non CHD, nonstroke) mortality, and nonvascular mortality. There was no evidence of a threshold within the range of usual fibrinogen level studied at any age. The age- and sex- adjusted hazard ratio per 1-g/L increase in usual fibrinogen level for CHD was 2.42 (95% confidence interval [CI], 2.24-2.60); stroke, 2.06 (95% CI, 1.83-2.33); other vascular mortality, 2.76 (95% CI, 2.28-3.35); and nonvascular mortality, 2.03 (95% CI, 1.90-2.18). The hazard ratios for CHD and stroke were reduced to about 1.8 after further adjustment for measured values of several established vascular risk factors. In a subset of 7011 participants with available C-reactive protein values, the findings for CHD were essentially unchanged following additional adjustment for C-reactive protein. The associations of fibrinogen level with CHD or stroke did not differ substantially according to sex, smoking, blood pressure, blood lipid levels, or several features of study design. CONCLUSIONS: In this large individual participant meta analysis, moderately strong associations were found between usual plasma fibrinogen level and the risks of CHD, stroke, other vascular mortality, and nonvascular mortality in a wide range of circumstances in healthy middle-aged adults. Assessment of any causal relevance of elevated fibrinogen levels to disease requires additional research. PMID- 16219888 TI - JAMA patient page. Palliative sedation. PMID- 16219889 TI - Comment: extensive prolongation of aPTT with argatroban in an elderly patient with improving renal function, normal hepatic enzymes, and metastatic lung cancer. PMID- 16219891 TI - HHS-CDC news: Direct and indirect effects of routine vaccination of children with 7-valent pneumococcal conjugate vaccine on incidence of invasive pneumococcal disease--US, 1998-2003. PMID- 16219892 TI - Thalidomide-associated thrombocytopenia. AB - OBJECTIVE: To report thrombocytopenia in a patient prescribed thalidomide for multiple myeloma (MM). CASE SUMMARY: A 70-year-old woman was diagnosed in 2003 with MM. At diagnosis, melphalan 0.25 mg/kg/day and prednisolone 2 mg/kg/day were started; however, the patient became refractory to therapy. Melphalan and prednisolone were discontinued, and monotherapy with dexamethasone 40 mg for 12 days per month was started. The patient's hematologic condition deteriorated again after about one year; dexamethasone was discontinued, and treatment with oral thalidomide 200 mg/day was initiated. About 2 weeks after thalidomide administration, the woman developed disabling adverse effects (flu-like symptoms, swollen fingers, rash and hematoma on her legs, shortness of breath, dry mouth, multiple petechiae). Laboratory testing showed neutropenia (neutrophils 0.4 x 10(9)/L) and thrombocytopenia (platelets 58 x 10(9)/L). Thalidomide was promptly discontinued; within 3 weeks, the laboratory values returned to pretreatment levels (1.3 x 10(9)/L and 267 x 10(9)/L, respectively) and her symptoms disappeared. DISCUSSION: Thrombocytopenia is a rarely reported hematologic adverse consequence of thalidomide therapy. A recent report identified 5 patients who developed thrombocytopenia while undergoing monotherapy with thalidomide for MM. According to the Naranjo probability scale, thalidomide was classified as the probable cause of thrombocytopenia in our patient. CONCLUSIONS: Unlike other antineoplastic drugs, thalidomide is rarely reported to cause severe hematologic toxicity. We present this case to increase clinicians' awareness for the potential of thalidomide to adversely affect platelet counts, particularly because its effectiveness in MM will likely result in expansion of its clinical use. PMID- 16219893 TI - Community pharmacy-based pharmaceutical care for asthma patients. AB - BACKGROUND: Despite significant progress in asthma drug therapy in recent years, there has been no major change in asthma morbidity and mortality. It is still important to determine whether pharmaceutical care (PC) influences health outcomes. OBJECTIVE: To evaluate the effectiveness of PC with regard to clinical, humanistic, and economic outcomes in adults with asthma. METHODS: An intervention study was conducted over 12 months. At baseline, 39 community/retail pharmacies, 84 primary care physicians (general practitioners, internal specialists, chest physicians), and 183 patients (aged 18-65 y) diagnosed with asthma were included. To evaluate economic outcomes, 2 German statutory health insurance funds provided 2 years of claims data for their insured patients (n = 55). A 1:10 matching was carried out to compare the data of this intervention subgroup with those of a control group (n = 550). RESULTS: Significant improvements were found for all humanistic outcomes (eg, asthma-specific quality of life, self-efficacy, knowledge, medication adherence). In addition, asthma severity, self-reported symptoms, peak expiratory flow, and patients' inhalation technique improved. Increases in forced expiratory volume in 1 second and vital capacity were not significant over time. Evaluation of the insurance claims data revealed a shift toward better adherence to evidence-based therapy. CONCLUSIONS: The study shows that PC for people with asthma has a positive impact on humanistic and, to some extent, on clinical outcomes. To determine potential economic benefits, future research should focus on patients with more severe asthma. PMID- 16219894 TI - Pharmacist impact on clinical outcomes in a diabetes disease management program via collaborative practice. AB - BACKGROUND: Clinical outcomes resulting from pharmaceutical care have been incompletely addressed in the diabetes population. We conducted a retrospective study evaluating clinical outcomes in a diabetes disease management program in which clinical pharmacists possessed collaborative practice agreements. OBJECTIVE: To evaluate changes in clinical outcomes for patients enrolled in a pharmacist-coordinated diabetes management program. METHODS: Medical records of 157 patients enrolled in the diabetes management program between June 2003 and April 2004 were retrospectively reviewed. Data collection included baseline and follow-up values for hemoglobin A(1C) (A1C) and lipids as well as frequency of adherence to preventive care, including annual foot and eye examinations and daily aspirin therapy. RESULTS: For patients with both baseline and follow-up data, the mean A1C reduction was 1.6% (n = 109; p < 0.001). For patients with an initial A1C of > or =8.5%, the mean reduction was 2.7% (n = 57; p < 0.001). The percentage of patients with A1C < or =7% increased from 19% at baseline to 50% at follow-up (p < 0.001). The mean low-density lipoprotein (LDL) reduction observed was 16 mg/dL (n = 73; NS) and the percentage of patients with LDL values < or =100 mg/dL increased from 30% at baseline to 56% at follow-up (p < 0.001). The frequency of microalbumin screening increased by 27% (p < 0.001), and the number of patients with annual eye and foot examinations increased by 27% (p < 0.05) and 15% (p < 0.05), respectively. The percentage of patients who had a positive microalbumin test and were taking a renal protective agent rose 19% from baseline to follow-up (NS). The percentage of patients taking daily aspirin increased from 42% at baseline to 80% at follow-up (p < 0.01). CONCLUSIONS: The pharmacist coordinated diabetes management program was effective in improving clinical markers for enrolled patients. Significant improvements were observed in A1C and LDL values as well as the frequency of adherence to preventive care. PMID- 16219895 TI - Possible improvement of neuroleptic-associated tardive dyskinesia during treatment with aripiprazole. PMID- 16219896 TI - The need for routine bleomycin test dosing in the 21st century. AB - OBJECTIVE: To review the clinical evidence for routine use of bleomycin test dosing. DATA SOURCES: English-language review articles, references from retrieved articles, case reports, and clinical trials were identified from a MEDLINE literature search (1966-July 2005). Key search terms included bleomycin, test dose, anaphylactic reactions, and hypersensitivity. Information from an unpublished E-mail survey, the manufacturer, and the Internet was also used. DATA SYNTHESIS: Early clinical trials and isolated case reports suggest that bleomycin induced acute hypersensitivity reactions occur in 1% of patients with lymphoma and <0.5% of those with solid tumors. The reactions are mainly characterized by high-grade fever, chills, hypotension, and in a few cases, cardiovascular collapse, which can lead to death. The exact mechanism of these reactions is unclear, but is thought to be related to the release of endogenous pyrogens from the host cells. Evidence does not suggest any correlation between doses and the onset or severity of the reactions. Supportive care, including hydration, steroids, antipyretics, and antihistamines, may resolve the symptoms. However, it may not completely prevent recurrences. CONCLUSIONS: The incidence of acute hypersensitivity or hyperpyrexic reactions associated with bleomycin is very low, but the reaction is potentially fatal. Clinicians should monitor their patients for any signs and symptoms of acute hyperpyrexic reactions during bleomycin administration. Since the onset of the reactions can occur with any dose of bleomycin and at any time, routine test dosing does not seem to predict when drug reactions may occur. PMID- 16219897 TI - Providing patient care in community pharmacies in Australia. AB - OBJECTIVE: To describe Australia's community pharmacy network in the context of the health system and outline the provision of services. DATA SYNTHESIS: The 5000 community pharmacies form a key component of the healthcare system for Australians, for whom health expenditures represent 9% of the Gross Domestic Product. A typical community pharmacy dispenses 880 prescriptions per week. Pharmacists are key partners in the Government's National Medicines Policy and contribute to its objectives through the provision of cognitive pharmaceutical services (CPS). The Third Community Pharmacy Agreement included funding for CPS including medication review and the provision of written drug information. Funding is also provided for a quality assurance platform with which the majority of pharmacies are accredited. Fifteen million dollars (Australian) have been allocated to research in community pharmacy, which has focused on achieving quality use of medicines (QUM), as well as developing new CPS and facilitating change. Elements of the Agreements have taken into account QUM principles and are now significant drivers of practice change. Although accounting for 10% of remuneration for community pharmacy, the provision of CPS represents a significant shift in focus to view pharmacy as a service provider. Delivery of CPS through the community pharmacy network provides sustainability for primary health care due to improvement in quality presumably associated with a reduction in healthcare costs. CONCLUSIONS: Australian pharmacy practice is moving strongly in the direction of CPS provision; however, change does not occur easily. The development of a change management strategy is underway to improve the uptake of professional and business opportunities in community pharmacy. PMID- 16219898 TI - Natalizumab for the treatment of multiple sclerosis and Crohn's disease. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, safety, and pivotal clinical trials for natalizumab in the treatment of multiple sclerosis (MS) and inflammatory bowel disease. DATA SOURCES: A PubMed/MEDLINE search was conducted (1966-June 2005), and information was obtained from Iowa Drug Information Services. Additional data sources included meeting abstracts, bibliographies from identified articles, and information from the manufacturer. STUDY SELECTION AND DATA EXTRACTION: Studies and review articles examining natalizumab were evaluated. All published, randomized clinical trials evaluating natalizumab in MS and IBD were included in this review. DATA SYNTHESIS: Natalizumab is the first drug in a new class of agents called selective adhesion molecule inhibitors. It has shown promising results in MS and inflammatory bowel disease and appears superior compared with current therapies in reducing relapse rates. However, 3 recent, confirmed case reports of progressive multifocal leukoencephalopathy (PML) create concern about natalizumab's use in combination with existing therapies or in undefined patient subgroups. Natalizumab was voluntarily withdrawn from the market in March 2005 while the drug's safety is further evaluated. CONCLUSIONS: Although long-term efficacy and safety of natalizumab have not been established, available data indicate that it is a novel drug for patients with MS or inflammatory bowel disease. Analysis of its possible association with PML will determine the risk-benefit evaluation and eventual place in therapy for natalizumab. PMID- 16219899 TI - Levosimendan: a novel inotropic agent for treatment of acute, decompensated heart failure. AB - OBJECTIVE: To review the literature on a novel calcium sensitizer, levosimendan. DATA SOURCES: Articles were identified through searches of MEDLINE (1966-June 2005), International Pharmaceutical Abstracts (1970-June 2005), and EMBASE (1992 June 2005) using the key words levosimendan, simendan, calcium sensitizer, calcium sensitiser, and congestive heart failure. STUDY SELECTION AND DATA EXTRACTION: Clinical trials and pharmacokinetic studies evaluating the safety and efficacy of levosimendan were selected. DATA SYNTHESIS: Levosimendan 6-24 mug/kg intravenous bolus followed by a 24-hour continuous infusion of 0.05-0.2 microg/kg/min improved cardiac output and reduced pulmonary capillary wedge pressure in a dose-dependent manner. Dose-ranging and randomized clinical trials have demonstrated improvement in symptoms and hemodynamics and short-term survival outcomes in the treatment of acute, decompensated heart failure. Clinical trials evaluating retrospective mortality data and combined endpoints (mortality, rehospitalization) have demonstrated better outcomes with levosimendan compared with dobutamine. The incidence of hypotension with levosimendan is not significantly different than with dobutamine, but there is a dose-related increase in heart rate. CONCLUSIONS: Levosimendan is useful in moderate to severe low-output heart failure in patients who have failed to respond to diuretics and vasodilators. Based on current studies, levosimendan appears to be a safe alternative to dobutamine for treatment of acute, decompensated heart failure. Prospective clinical trials are needed to confirm the effect of levosimendan on long-term survival and its role in heart failure in the setting of myocardial infarction. PMID- 16219900 TI - Incremental effects of concurrent pharmacotherapeutic regimens for heart failure on hospitalizations and costs. AB - BACKGROUND: Inappropriate medication use in patients with heart failure (HF) presents challenges in providing optimal, evidence-based care. OBJECTIVE: To evaluate the incremental differences of concurrent and persistent use of angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, loop diuretics, and digoxin on the one-year, all-cause risk of hospitalization and total healthcare costs associated with treatment of HF in patients enrolled in a managed care organization within the US. METHODS: A retrospective database analysis was conducted spanning from January 1, 1997, to December 31, 1999. Multivariate regression methods were used to examine the association between treatment regimens and hospitalizations or costs after controlling for patient demographics and risk factors. RESULTS: Of the 1903 patients meeting inclusion criteria, 32.3% (n = 615) received none of the 4 HF agents studied and were associated with a 2.5 times greater risk (p < or = 0.001) of hospitalization and 43.6% higher (p < or = 0.001) total costs compared with all other patients with HF. Comparatively, 13.9% (n = 264) utilized the HF medications investigated for at least 6 months. Of those with persistent use of > or =3 agents, approximate decreases in hospitalizations were noted of 80% (p < or = 0.001) and total costs of 70% (p < or = 0.001) relative to patients receiving no HF therapy. CONCLUSIONS: A substantial portion of patients with HF may be receiving suboptimal pharmacotherapeutic care in real-world practice settings, potentially incurring large increases in hospitalizations and total costs. Quality improvement initiatives should seek to identify and manage those not being treated according to guideline recommendations. PMID- 16219901 TI - Open trial of pindolol in the treatment of fibromyalgia. AB - BACKGROUND: Evidence suggests that fibromyalgia is related to both chronic sympathetic hyperactivity and decreased levels of serotonin. OBJECTIVE: To examine the efficacy of pindolol, a mixed serotonin (5-HT)(1A) presynaptic autoreceptor/beta-adrenergic receptor antagonist, in the treatment of fibromyalgia. METHODS: An open trial was conducted using 20 female patients who met the American College of Rheumatology criteria for fibromyalgia. Treatment was initiated with pindolol 7.5 mg/day and titrated to a maximum dose of 15 mg/day for a total of 90 days. Primary outcome measures were tender point analysis and the Fibromyalgia Impact Questionnaire (FIQ). Anxiety and depression were measured with the Hamilton Depression and Anxiety Scales and Beck Depression Inventory. RESULTS: There was significant improvement in primary outcome measures, including Tender Point Count (mean +/- SD, 16.3 +/- 2.2 vs 12.3 +/- 5.0; F = 8.9; p < 0.001), Tender Point Score (24.4 +/- 5.7 vs 17.5 +/- 9.4; F = 7.8; p < 0.001), and FIQ (45.3 +/- 10.8 vs 35.0 +/- 15.0; F = 5.6; p < 0.005). The depression and anxiety scores did not change significantly among women who completed the study, while the impact on cardiovascular parameters was clinically insignificant. CONCLUSIONS: While the current results are encouraging, further studies are needed to determine whether pindolol might be effective in the treatment of fibromyalgia. Limitations of this study include small group size and lack of placebo control. PMID- 16219902 TI - Identification of inappropriate drug prescribing by computerized, retrospective DUR screening in Korea. AB - BACKGROUND: In Korea, the drug use process has changed significantly since the new pharmacy law was implemented in 2000, separating the prescribing and dispensing functions between physicians and pharmacists and mandating prospective drug use review (DUR) practice by pharmacists immediately before dispensing medications. However, a high prevalence of inappropriate prescribing has been suspected by the public, pharmacists, and health insurance managers, possibly due to suboptimal DUR practice by pharmacists. OBJECTIVE: To assess overall patterns of drug usage and potential problems of inappropriate use in outpatient settings by analyzing prescription data that were electronically submitted to the national health insurance manager with a computerized DUR system and develop a computerized adjudication system model for drug claims. METHODS: The national prescription drug claims data that were submitted electronically by pharmacies located in the northern part of Korea during 15 days in 2002 were retrospectively screened against the predetermined DUR standards of the selected criteria on drug dosage, duration of therapy, and drug interaction using the DUR screening system. The results of all the DUR conflicts were further validated manually by an expert panel and statistically analyzed to determine drug use patterns. RESULTS: Of 31,994,260 drugs prescribed, 3,325,760 (10.4%) items showed a conflict with at least one of the DUR standards. The average number of drugs prescribed on each prescription was 4.07, and even more troubling was the high incidence of under dosing, over-dosing, and contraindicated drug prescribing. CONCLUSIONS: It is evident that inappropriate drug prescribing is very common in Korea; thus, a great deal of attention is urgently needed in the country with respect to proper prescribing and supportive interventions. PMID- 16219903 TI - Aralast: a new alpha1-protease inhibitor for treatment of alpha-antitrypsin deficiency. AB - OBJECTIVE: To review the epidemiology, pathogenesis, and management of patients with alpha-antitrypsin (AAT) deficiency syndrome and compare Aralast with Prolastin, 2 of the 3 available human plasma-derived AAT agents. DATA SOURCES: Articles were identified using a MEDLINE (1966-September 2005) search with MESH headings that included alpha-antitrypsin and emphysema. STUDY SELECTION AND DATA EXTRACTION: All papers from peer-reviewed journals on the laboratory or clinical efficacy of plasma-derived AAT (eg, Prolastin, Aralast) for patients with this autosomal recessive disorder were reviewed. DATA SYNTHESIS: Clinical trials found that AAT augmentation prevents progression of AAT-deficient emphysema and thus its associated morbidity and mortality. Treatment with Aralast has been shown to be safe and well tolerated, with a low incidence of mild to moderate adverse events. Pharmacoeconomics studies of AAT augmentation demonstrated that the use of Aralast was cost-effective as lifelong augmentation therapy for AAT-deficient emphysema. CONCLUSIONS: Because of its effectiveness and extra safety measure compared with Prolastin, Aralast should be recommended for formulary inclusion. PMID- 16219904 TI - Cell signaling: from beginning to end. AB - The Connections Maps database and the graphical Web interface at Science's STKE for accessing the data represent an emerging Web-based tool through which researchers in cell signaling can share their knowledge with the scientific community. This tool provides a mechanism by which researchers, students, and clinicians can transform information about interactions between biomolecules into knowledge about a cellular process. In 2005, new Pathways were added to the Connections Maps database and several of the existing Pathways were updated. PMID- 16219905 TI - Curcumin (diferuloylmethane) down-regulates expression of cell proliferation and antiapoptotic and metastatic gene products through suppression of IkappaBalpha kinase and Akt activation. AB - Curcumin (diferuloylmethane), an anti-inflammatory agent used in traditional medicine, has been shown to suppress cellular transformation, proliferation, invasion, angiogenesis, and metastasis through a mechanism not fully understood. Because several genes that mediate these processes are regulated by nuclear factor-kappaB (NF-kappaB), we have postulated that curcumin mediates its activity by modulating NF-kappaB activation. Indeed, our laboratory has shown previously that curcumin can suppress NF-kappaB activation induced by a variety of agents (J Biol Chem 270:24995-50000, 1995). In the present study, we investigated the mechanism by which curcumin manifests its effect on NF-kappaB and NF-kappaB regulated gene expression. Screening of 20 different analogs of curcumin showed that curcumin was the most potent analog in suppressing the tumor necrosis factor (TNF)-induced NF-kappaB activation. Curcumin inhibited TNF-induced NF-kappaB dependent reporter gene expression in a dose-dependent manner. Curcumin also suppressed NF-kappaB reporter activity induced by tumor necrosis factor receptor (TNFR)1, TNFR2, NF-kappaB-inducing kinase, IkappaB kinase complex (IKK), and the p65 subunit of NF-kappaB. Such TNF-induced NF-kappaB-regulated gene products involved in cellular proliferation [cyclooxygenase-2 (COX-2), cyclin D1, and c myc], antiapoptosis [inhibitor of apoptosis protein (IAP)1, IAP2, X-chromosome linked IAP, Bcl-2, Bcl-x(L), Bfl-1/A1, TNF receptor-associated factor 1, and cellular Fas-associated death domain protein-like interleukin-1beta-converting enzyme inhibitory protein-like inhibitory protein], and metastasis (vascular endothelial growth factor, matrix metalloproteinase-9, and intercellular adhesion molecule-1) were also down-regulated by curcumin. COX-2 promoter activity induced by TNF was abrogated by curcumin. We found that curcumin suppressed TNF-induced nuclear translocation of p65, which corresponded with the sequential suppression of IkappaBalpha kinase activity, IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 phosphorylation, p65 nuclear translocation, and p65 acetylation. Curcumin also inhibited TNF-induced Akt activation and its association with IKK. Glutathione and dithiothreitol reversed the effect of curcumin on TNF-induced NF kappaB activation. Overall, our results indicated that curcumin inhibits NF kappaB activation and NF-kappaB-regulated gene expression through inhibition of IKK and Akt activation. PMID- 16219906 TI - Synaptic expression of glutamate receptor after encoding of fear memory in the rat amygdala. AB - Fear conditioning has been ascribed to presynaptic mechanisms, particularly presynaptic facilitation of transmission at thalamo- and cortico-amygdala synapses. Here, by labeling surface receptors with biotin or using membrane fractionation approaches, we report that fear conditioning resulted in an increase in surface expression of GluR1 subunit of alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid receptors in the amygdala, whereas total GluR1 mRNA and protein levels were unchanged. The control group that received conditioned stimulus (CS) and unconditioned stimulus in an unpaired fashion did not present any increase, indicating that GluR1 increase was specific to the learning component of the task. Conditioning-induced increase in surface expression of GluR1 depended on the activation of N-methyl-d-aspartate receptors and protein kinases and required the synthesis of new proteins. CS-alone trials applied 24 h before training attenuated fear-potentiated startle and prevented conditioning induced increase in surface expression of GluR1. Increase in GluR1 was also observed in the amygdala slices after delivery of tetanic stimulation that elicited long-term potentiation of synaptic transmission. Proteasome inhibitor increased surface expression of GluR1 in a time- and dose-dependent manner. Furthermore, pretraining administration of proteasome inhibitor into the amygdala facilitated the fear-potentiated startle. These results suggest that long-term memory formation is correlated with the change in synaptic expression of GluR1, and trafficking of GluR1 to the synaptic sites contributes at least in part to the expression of fear memory. PMID- 16219907 TI - Binding site flexibility: molecular simulation of partial and full agonists within a glutamate receptor. AB - Ionotropic glutamate receptors mediate fast synaptic transmission in the mammalian central nervous system and play an important role in many different functions, including memory and learning. They have also been implicated in a variety of neuropathologies and as such have generated widespread interest in their structure and function. Molecular Dynamics simulations (5 x 20 ns) of the ligand-binding core of the GluR2 glutamate receptor were performed. Through simulations of both wild type and the L650T mutant, we show that the degree of protein flexibility can be correlated with the extent to which the binding cleft is open. In agreement with recent experiments, the simulations of kainate with the wild-type construct show a slight increase in beta-sheet content that we are able to localize to two specific regions. During one simulation, the protein made a transition from an open-cleft conformation to a closed-cleft conformation. This closed cleft conformation closely resembles the closed-cleft crystal structure, thus indicating a potential pathway for conformational change associated with receptor activation. Analysis of the binding pocket suggests that partial agonists possess a greater degree of flexibility within the pocket that may help to explain why they are less efficient at opening the channel than full agonists. Examination of water molecules surrounding the ligands reveals that mobility in distinct subsites can be a discriminator between full and partial agonism and will be an important consideration in the design of drugs against these receptors. PMID- 16219908 TI - Flavopiridol and histone deacetylase inhibitors promote mitochondrial injury and cell death in human leukemia cells that overexpress Bcl-2. AB - Interactions between the cyclin-dependent kinase (CDK) inhibitor flavopiridol and histone deacetylase (HDAC) inhibitors (suberoylanilide hydroxamide and sodium butyrate) were examined in human leukemia cells (U937 and HL-60) ectopically expressing Bcl-2/Bcl-x(L) and in primary AML cells. Coadministration of flavopiridol with HDAC inhibitors synergistically potentiated mitochondrial damage (cytochrome c, second mitochondria-derived activator of caspases/direct IAP binding protein with low pI, and apoptosis-inducing factor release), caspase activation, poly(ADP-ribose) polymerase degradation, and cell death in both wild type and Bcl-2- or Bcl-x(L)-overexpressing cells and induced a pronounced loss of clonogenicity. In contrast, Bcl-2 and Bcl-x(L) largely blocked these events in cells exposed to the cytotoxic agent 1-beta-d-arabinofuranosylcytosine (ara-C). Enforced expression of dominant-negative Fas-associated death domain failed to protect cells from the flavopiridol/histone deacetylase inhibitor (HDACI) regimen, arguing against the involvement of the receptor pathway in lethality. Ectopic expression of a phosphorylation loop-deleted Bcl-2 or Bcl-2 lacking the serine(70) phosphorylation site, which dramatically protected cells from ara-C lethality, delayed but did not prevent flavopiridol/HDAC inhibitor-induced mitochondrial injury, cell death, or loss of clonogenicity. Ectopic expression of Bcl-2 or Bcl-x(L) was also unable to prevent the flavopiridol/HDACI regimen from inducing a conformational change in and mitochondrial translocation of Bax, and it did not attenuate Bax dimerization. As a whole, these findings indicate that in contrast to certain conventional cytotoxic agents such as ara-C, overexpression of Bcl-2 or Bcl-x(L) are largely ineffective in preventing perturbations in Bax, mitochondrial injury, and cell death in human leukemia cells subjected to simultaneous CDK and HDAC inhibition. They also raise the possibility that a strategy combining CDK and HDAC inhibitors may be effective against drug-resistant leukemia cells overexpressing Bcl-2 or Bcl-x(L). PMID- 16219910 TI - Activation of human ether-a-go-go-related gene potassium channels by the diphenylurea 1,3-bis-(2-hydroxy-5-trifluoromethyl-phenyl)-urea (NS1643). AB - The cardiac action potential is generated by a concerted action of different ion channels and transporters. Dysfunction of any of these membrane proteins can give rise to cardiac arrhythmias, which is particularly true for the repolarizing potassium channels. We suggest that an increased repolarization current could be a new antiarrhythmic principle, because it possibly would attenuate afterdepolarizations, ischemic leak currents, and reentry phenomena. Repolarization of the cardiac myocytes is crucially dependent on the late rapid delayed rectifier current (I(Kr)) conducted by ether-a-go-go-related gene (ERG) potassium channels. We have developed the diphenylurea compound 1,3-bis-(2 hydroxy-5-trifluoromethyl-phenyl)-urea (NS1643) and tested whether this small organic molecule could increase the activity of human ERG (HERG) channels expressed heterologously. In Xenopus laevis oocytes, NS1643 increased both steady state and tail current at all voltages tested. The EC(50) value for HERG channel activation was 10.5 microM. These results were reproduced on HERG channels expressed in mammalian human embryonic kidney 293 cells. In guinea pig cardiomyocytes, studied by patch clamp, application of 10 microM NS1643 activated I(Kr) and significantly decreased the action potential duration to 65% of the control values. The effect could be reverted by application of the specific HERG channel inhibitor 4'-[[1-[2-(6-methyl-2-pyridyl)ethyl]-4-piperidinyl]carbonyl] methanesulfonanilide (E-4031) at 100 nM. Application of NS1643 also resulted in a prolonged postrepolarization refractory time. Finally, cardiomyocytes exposed to NS1643 resisted reactivation by small depolarizing currents mimicking early afterdepolarizations. In conclusion, HERG channel activation by small molecules such as NS1643 increases the repolarization reserve and presents an interesting new antiarrhythmic approach. PMID- 16219909 TI - A novel drug binding site on voltage-gated sodium channels in rat brain. AB - The effectiveness of several antiepileptic, analgesic, and neuroprotective drugs is attributable to state-dependent inhibition of voltage-gated sodium channels. To help characterize their site and mode of action on sodium channels, a member of the lamotrigine family, R-(-)-2,4-diamino-6-(fluromethyl)-5-(2,3,5 trichlorophenyl)-pyrimidine (BW202W92), was radiolabeled and used as a binding ligand in rat forebrain synaptosomes. Although the level of specific [(3)H]BW202W92 binding in a standard incubation medium was relatively poor, low concentrations of tetrodotoxin (EC(50) = 2-3 nM) greatly enhanced the binding, apparently by increasing the affinity of the binding sites. Tetrodotoxin dependent binding was stereoselective (the less active enantiomer, S-(-)-2,4 diamino-6-(fluromethyl)-5-(2,3,5-trichlorophenyl)-pyrimidine (BW203W92), was up to 30-fold less potent, depending on conditions) and was extremely sensitive to inhibition by raised K(+) concentration (IC(50) = 5.9 mM), an effect that was ascribed to changes in membrane potential. In addition, the binding was inhibited by sodium channel neurotoxins acting on sites 3 and 4, but it was resistant to batrachotoxin (site 2) and brevetoxin (site 5). Several drugs acting on sodium channels displaced tetrodotoxin-dependent [(3)H]BW202W92 binding, and most of those tested showed different affinities under depolarized (100 mM K(+)) and polarized (1 mM K(+)) conditions. In a subset of compounds for which data were available, binding affinity in depolarized synaptosomes correlated well with apparent affinity for the inactivated state of sodium channels. The [(3)H]BW202W92 binding site is novel and is likely to represent a pharmacologically important site of action of drugs on voltage-gated sodium channels in the brain. PMID- 16219911 TI - The blockage of survivin and securin expression increases the cytochalasin B induced cell death and growth inhibition in human cancer cells. AB - Survivin and securin proteins are overexpressed in most cancer cells that have been shown to regulate mitotic progression. In this study, we investigated the roles of survivin and securin on cytochalasin B, a cytokinesis blocker mediating the cytotoxicity and cell growth inhibition in human cancer cells. The human lung carcinoma cell lines A549 and H1299 highly expressed survivin proteins in mitosis and concentrated on the midbodies during cytokinesis. Cytochalasin B significantly decreased cell survival, inhibited cell growth, increased the levels of G(2)/M fractions, and induced binuclei formation in lung carcinoma cells; however, the survivin proteins were concentration-dependently increased by 1 to 5 mug/ml cytochalasin B for 24 h. It is noteworthy that the expression of securin proteins was decreased in cytochalasin B-treated lung carcinoma cells. Transfection of 20 to 40 nM survivin siRNA for 48 h significantly induced the formation of multiple nuclei and apoptosis but decreased the levels of survivin and securin proteins in A549 cells. Cotreatment with survivin small interfering RNA (siRNA) and cytochalasin B increased the cytotoxicity and cell growth inhibition. In addition, the securin-null colorectal carcinoma cells were more susceptible to the cytotoxicity after cytochalasin B and survivin siRNA treatments than the securin-wild-type cells. As a whole, our results indicate that the inhibition of survivin and securin protein expression may increase the cell death and growth inhibition after cytochalasin B treatment in human cancer cells. PMID- 16219912 TI - Regulation and binding of pregnane X receptor by nuclear receptor corepressor silencing mediator of retinoid and thyroid hormone receptors (SMRT). AB - The pregnane X receptor (PXR) is an orphan nuclear receptor predominantly expressed in liver and intestine. PXR coordinates hepatic responses to prevent liver injury induced by environmental toxins. PXR activates cytochrome P450 3A4 gene expression upon binding to rifampicin (Rif) and clotrimazole (CTZ) by recruiting transcriptional coactivators. It remains unclear whether and how PXR regulates gene expression in the absence of ligand. In this study, we analyzed interactions between PXR and the silencing mediator of retinoid and thyroid hormone receptors (SMRT) and determined the role of SMRT in regulating PXR activity. We show that SMRT interacts with PXR in glutathione S-transferase pull down, yeast two-hybrid, and mammalian two-hybrid assays. The interaction is mediated through the ligand-binding domain of PXR and the SMRTs' nuclear receptor interacting domain 2. The PXR-SMRT interaction is sensitive to species-specific ligands, and Rif causes an exchange of the corepressor SMRT with the p160 coactivator known as receptor-associated coactivator 3 (RAC3). Deletion of the PXR's activation function 2 helix enhances SMRT binding and abolishes ligand dependent dissociation of SMRT. Coexpression of PXR with SMRT results in colocalization at discrete nuclear foci. Finally, transient transfection assays show that overexpression of SMRT inhibits PXR's transactivation of the Cyp3A4 promoter, whereas silencing of SMRT enhances the reporter expression. Taken together, our results suggest that the corepressor SMRT may bind to and regulate the transcriptional activity of PXR. PMID- 16219913 TI - Renal medullary tissue oxygenation is dependent on both cortical and medullary blood flow. AB - The aim of the current study was to determine whether renal medullary oxygenation is independent of the level of cortical blood flow by testing responses to stimuli that selectively reduce blood flow in either the cortex or medulla. In anesthetized rabbits, renal arterial infusion of [Phe(2),Ile(3),Orn(8)] vasopressin selectively reduced medullary perfusion and Po(2) (-54 +/- 24 and -50 +/- 10%, respectively) but did not significantly affect cortical perfusion or tissue oxygenation. In contrast, stimulation of the renal nerves resulted in renal cortical ischemia with reductions in total renal blood flow (-76 +/- 3% at 4 Hz), cortical perfusion (-57 +/- 17%), and cortical Po(2) (-44 +/- 12%). Medullary tissue Po(2) was reduced by -70 +/- 5% at 4 Hz, despite medullary perfusion being unaffected and distal tubular sodium reabsorption being reduced (by -83.3 +/- 1.2% from baseline). In anesthetized rats, in which renal perfusion pressure was maintained with an aortic constrictor, intravenous infusion of ANG II (0.5-5 microg. kg(-1).min(-1)) dose dependently reduced cortical perfusion (up to -65 +/- 3%; P < 0.001) and cortical Po(2) (up to -57 +/- 4%; P < 0.05). However, medullary perfusion was only significantly reduced at the highest dose (5 microg. kg(-1).min(-1); by 29 +/- 6%). Medullary perfusion was not reduced by 1 microg. kg(-1).min(-1) ANG II, but medullary Po(2) was significantly reduced ( 12 +/- 4%). Thus, although cortical and medullary blood flow may be independently regulated, medullary oxygenation may be compromised during moderate to severe cortical ischemia even when medullary blood flow is maintained. PMID- 16219914 TI - Role of deadenylation and AUF1 binding in the pH-responsive stabilization of glutaminase mRNA. AB - During chronic metabolic acidosis, increased expression of renal glutaminase (GA) results from selective stabilization of the GA mRNA. This response is mediated by a direct repeat of an 8-base adenylate-uridylate (AU) sequence that binds zeta crystallin and functions as a pH response element (pH-RE). A tetracycline responsive promoter system was developed in LLC-PK(1)-F(+) cells to perform pulse chase analysis of the turnover of a chimeric beta-globin (betaG) mRNA that contains 960 bp of the 3'-UTR of GA mRNA including the pH-RE. The betaG-GA mRNA exhibits a 14-fold increase in half-life when the LLC-PK(1)-F(+) cells are transferred to acidic medium. RNase H cleavage and Northern blot analysis of the 3'-ends established that rapid deadenylation occurred concomitantly with the rapid decay of the betaG-GA mRNA in cells grown in normal medium. Stabilization of the betaG-GA mRNA in acidic medium is associated with a pronounced decrease in the rate of deadenylation. Mutation of the pH-RE within the betaG-GA mRNA blocked the pH-responsive stabilization, but not the rapid decay, whereas insertion of only a 29-bp segment containing the pH-RE was sufficient to produce both a rapid decay and a pH-responsive stabilization. Various kidney cells express multiple isoforms of AUF1, an AU-binding protein that enhances mRNA turnover. RNA gel shift assays demonstrated that the recombinant p40 isoform of AUF1 binds to the pH-RE with high affinity and specificity. Thus AUF1 may mediate the rapid turnover of the GA mRNA, whereas increased binding of zeta-crystallin during acidosis may inhibit degradation and result in selective stabilization. PMID- 16219915 TI - Interactions between TGF-dependent and myogenic oscillations in tubular pressure and whole kidney blood flow in both SDR and SHR. AB - We previously showed that nonlinear interactions between the two renal autoregulatory mechanics (tubuloglomerular feedback and the myogenic mechanism) were observed in the stop flow pressure (SFP) and whole kidney blood flow data from Sprague-Dawley rats (SDR) using time-invariant bispectrum analysis (3, 4). No such nonlinear interactions were observed in either SFP or whole kidney blood flow data obtained from spontaneously hypertensive rats (SHR). We speculated that the failure to detect nonlinear interactions in the SHR data may be related to our observation that these interactions were not continuous and therefore had time-varying characteristics. Thus the absence of such nonlinear interactions may be due to an inappropriate time-invariant method being applied to data that are especially time varying in nature. We examine this possibility in this paper by using a time-varying bispectrum approach, which we developed for this purpose. Indeed, we found significant nonlinear interactions in SHR (n = 18 for SFP; n = 12 for whole kidney blood flow). Moreover, the duration of nonlinear coupling is found statistically to be longer (P = 0.001) in SFP data from either SDR or SHR than it is in whole kidney data from either type of rat. We conclude that nonlinear coupling is present at both the single nephron as well as the whole kidney level for SDR and SHR. In addition, SHR data at the whole kidney level exhibit the most transient nonlinear coupling phenomena. PMID- 16219916 TI - Gene expression of 5-, 12-, and 15-lipoxygenases and leukotriene receptors along the rat nephron. AB - The arachidonate signaling pathways comprise prostanoids formed by cyclooxygenases, EETs, and HETEs formed by cytochrome P-450 (CYP) enzymes and HETEs and leukotrienes generated by lipoxygenases. Whereas the intrarenal localization of cyclooxygenases and of some CYP enzymes along the nephron has already been determined, the localization of lipoxygenases and leukotriene forming enzymes together with leukotriene receptors in the kidney is less clear. This study therefore aimed to determine the expression of 5-, 12-, and 15 lipoxygenases as well as the leukotriene receptors along the rat nephron. The kidneys were dissected into cortex and outer and inner medulla, and the microdissected nephron segments were collected after a collagenase digestion. mRNA abundance was determined by RT-PCR and real-time PCR. 15-LOX mRNA showed a characteristic expression pattern along the distal nephron. 12-LOX mRNA was only found in the glomerulus. Similarly, 5-LOX mRNAs together with 5-LOX-activating protein mRNAs were expressed in the glomerulus and also in the vasa recta. The leukotriene A4 hydrolase was found in all nephron segments, whereas leukotriene C4 synthase mRNA could not be found in any nephron segment. The leukotriene receptor B4 and the cysteinyl leukotriene receptor type 1 were selectively expressed in the glomerulus, whereas cysteinyl receptor type 2 was not found in any nephron segment. Our data suggest that the glomerulus is a major source and target for 5- and 12-HETE and for leukotrienes. The collecting duct system, on the other hand, appears to be a major source of 15-HETE. PMID- 16219917 TI - High impact. PMID- 16219918 TI - Regulatory functions of phospholipase D and phosphatidic acid in plant growth, development, and stress responses. PMID- 16219921 TI - Comparative plant genomics resources at PlantGDB. AB - PlantGDB (http://www.plantgdb.org/) is a database of plant molecular sequences. Expressed sequence tag (EST) sequences are assembled into contigs that represent tentative unique genes. EST contigs are functionally annotated with information derived from known protein sequences that are highly similar to the putative translation products. Tentative Gene Ontology terms are assigned to match those of the similar sequences identified. Genome survey sequences are assembled similarly. The resulting genome survey sequence contigs are matched to ESTs and conserved protein homologs to identify putative full-length open reading frame containing genes, which are subsequently provisionally classified according to established gene family designations. For Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa), the exon-intron boundaries for gene structures are annotated by spliced alignment of ESTs and full-length cDNAs to their respective complete genome sequences. Unique genome browsers have been developed to present all available EST and cDNA evidence for current transcript models (for Arabidopsis, see the AtGDB site at http://www.plantgdb.org/AtGDB/; for rice, see the OsGDB site at http://www.plantgdb.org/OsGDB/). In addition, a number of bioinformatic tools have been integrated at PlantGDB that enable researchers to carry out sequence analyses on-site using both their own data and data residing within the database. PMID- 16219919 TI - Characterizing the grape transcriptome. Analysis of expressed sequence tags from multiple Vitis species and development of a compendium of gene expression during berry development. AB - We report the analysis and annotation of 146,075 expressed sequence tags from Vitis species. The majority of these sequences were derived from different cultivars of Vitis vinifera, comprising an estimated 25,746 unique contig and singleton sequences that survey transcription in various tissues and developmental stages and during biotic and abiotic stress. Putatively homologous proteins were identified for over 17,752 of the transcripts, with 1,962 transcripts further subdivided into one or more Gene Ontology categories. A simple structured vocabulary, with modules for plant genotype, plant development, and stress, was developed to describe the relationship between individual expressed sequence tags and cDNA libraries; the resulting vocabulary provides query terms to facilitate data mining within the context of a relational database. As a measure of the extent to which characterized metabolic pathways were encompassed by the data set, we searched for homologs of the enzymes leading from glycolysis, through the oxidative/nonoxidative pentose phosphate pathway, and into the general phenylpropanoid pathway. Homologs were identified for 65 of these 77 enzymes, with 86% of enzymatic steps represented by paralogous genes. Differentially expressed transcripts were identified by means of a stringent believability index cutoff of > or =98.4%. Correlation analysis and two dimensional hierarchical clustering grouped these transcripts according to similarity of expression. In the broadest analysis, 665 differentially expressed transcripts were identified across 29 cDNA libraries, representing a range of developmental and stress conditions. The groupings revealed expected associations between plant developmental stages and tissue types, with the notable exception of abiotic stress treatments. A more focused analysis of flower and berry development identified 87 differentially expressed transcripts and provides the basis for a compendium that relates gene expression and annotation to previously characterized aspects of berry development and physiology. Comparison with published results for select genes, as well as correlation analysis between independent data sets, suggests that the inferred in silico patterns of expression are likely to be an accurate representation of transcript abundance for the conditions surveyed. Thus, the combined data set reveals the in silico expression patterns for hundreds of genes in V. vinifera, the majority of which have not been previously studied within this species. PMID- 16219920 TI - Combining experimental and predicted datasets for determination of the subcellular location of proteins in Arabidopsis. AB - Substantial experimental datasets defining the subcellular location of Arabidopsis (Arabidopsis thaliana) proteins have been reported in the literature in the form of organelle proteomes built from mass spectrometry data (approximately 2,500 proteins). Subcellular location for specific proteins has also been published based on imaging of chimeric fluorescent fusion proteins in intact cells (approximately 900 proteins). Further, the more diverse history of biochemical determination of subcellular location is stored in the entries of the Swiss-Prot database for the products of many Arabidopsis genes (approximately 1,800 proteins). Combined with the range of bioinformatic targeting prediction tools and comparative genomic analysis, these experimental datasets provide a powerful basis for defining the final location of proteins within the wide variety of subcellular structures present inside Arabidopsis cells. We have analyzed these published experimental and prediction data to answer a range of substantial questions facing researchers about the veracity of these approaches to determining protein location and their interrelatedness. We have merged these data to form the subcellular location database for Arabidopsis proteins (SUBA), providing an integrated understanding of protein location, encompassing the plastid, mitochondrion, peroxisome, nucleus, plasma membrane, endoplasmic reticulum, vacuole, Golgi, cytoskeleton structures, and cytosol (www.suba.bcs.uwa.edu.au). This includes data on more than 4,400 nonredundant Arabidopsis protein sequences. We also provide researchers with an online resource that may be used to query protein sets or protein families and determine whether predicted or experimental location data exist; to analyze the nature of contamination between published proteome sets; and/or for building theoretical subcellular proteomes in Arabidopsis using the latest experimental data. PMID- 16219922 TI - GERMINATE. a generic database for integrating genotypic and phenotypic information for plant genetic resource collections. AB - The extensive germplasm resource collections that are now available for major crop plants and their wild relatives will increasingly provide valuable biological and bioinformatics resources for plant physiologists and geneticists to dissect the molecular basis of key traits and to develop highly adapted plant material to sustain future breeding programs. A key to the efficient deployment of these resources is the development of information systems that will enable the collection and storage of biological information for these plant lines to be integrated with the molecular information that is now becoming available through the use of high-throughput genomics and post-genomics technologies. The GERMINATE database has been designed to hold a diverse variety of data types, ranging from molecular to phenotypic, and to allow querying between such data for any plant species. Data are stored in GERMINATE in a technology-independent manner, such that new technologies can be accommodated in the database as they emerge, without modification of the underlying schema. Users can access data in GERMINATE databases either via a lightweight Perl-CGI Web interface or by the more complex Genomic Diversity and Phenotype Connection software. GERMINATE is released under the GNU General Public License and is available at http://germinate.scri.sari.ac.uk/germinate/. PMID- 16219924 TI - The International Rice Information System. A platform for meta-analysis of rice crop data. AB - Ambiguous germplasm identification; difficulty in tracing pedigree information; and lack of integration between genetic resources, characterization, breeding, evaluation, and utilization data are constraints in developing knowledge intensive crop improvement programs. To address these constraints, the International Crop Information System (www.icis.cgiar.org), a database system for the management and integration of global information on genetic resources and crop improvement for any crop, was developed by genetic resource specialists, crop scientists, and information technicians associated with the Consultative Group for International Agricultural Research and collaborative partners. The International Rice Information System (www.iris.irri.org) is the rice (Oryza species) implementation of the International Crop Information System. New components are now being added to the International Rice Information System to handle the diversity of rice functional genomics data including genomic sequence data, molecular genetic data, expression data, and proteomic information. Users access information in the database through stand-alone programs and Web interfaces, which offer specialized applications and customized views to researchers with different interests. PMID- 16219923 TI - Plant-based microarray data at the European Bioinformatics Institute. Introducing AtMIAMExpress, a submission tool for Arabidopsis gene expression data to ArrayExpress. AB - ArrayExpress is a public microarray repository founded on the Minimum Information About a Microarray Experiment (MIAME) principles that stores MIAME-compliant gene expression data. Plant-based data sets represent approximately one-quarter of the experiments in ArrayExpress. The majority are based on Arabidopsis (Arabidopsis thaliana); however, there are other data sets based on Triticum aestivum, Hordeum vulgare, and Populus subsp. AtMIAMExpress is an open-source Web-based software application for the submission of Arabidopsis-based microarray data to ArrayExpress. AtMIAMExpress exports data in MAGE-ML format for upload to any MAGE ML-compliant application, such as J-Express and ArrayExpress. It was designed as a tool for users with minimal bioinformatics expertise, has comprehensive help and user support, and represents a simple solution to meeting the MIAME guidelines for the Arabidopsis community. Plant data are queryable both in ArrayExpress and in the Data Warehouse databases, which support queries based on gene-centric and sample-centric annotation. The AtMIAMExpress submission tool is available at http://www.ebi.ac.uk/at-miamexpress/. The software is open source and is available from http://sourceforge.net/projects/miamexpress/. For information, contact miamexpress@ebi.ac.uk. PMID- 16219925 TI - GrainGenes 2.0. an improved resource for the small-grains community. AB - GrainGenes (http://wheat.pw.usda.gov) is an international database for genetic and genomic information about Triticeae species (wheat [Triticum aestivum], barley [Hordeum vulgare], rye [Secale cereale], and their wild relatives) and oat (Avena sativa) and its wild relatives. A major strength of the GrainGenes project is the interaction of the curators with database users in the research community, placing GrainGenes as both a data repository and information hub. The primary intensively curated data classes are genetic and physical maps, probes used for mapping, classical genes, quantitative trait loci, and contact information for Triticeae and oat scientists. Curation of these classes involves important contributions from the GrainGenes community, both as primary data sources and reviewers of published data. Other partially automated data classes include literature references, sequences, and links to other databases. Beyond the GrainGenes database per se, the Web site incorporates other more specific databases, informational topics, and downloadable files. For example, unique BLAST datasets of sequences applicable to Triticeae research include mapped wheat expressed sequence tags, expressed sequence tag-derived simple sequence repeats, and repetitive sequences. In 2004, the GrainGenes project migrated from the AceDB database and separate Web site to an integrated relational database and Internet resource, a major step forward in database delivery. The process of this migration and its impacts on database curation and maintenance are described, and a perspective on how a genomic database can expedite research and crop improvement is provided. PMID- 16219926 TI - Fatigue and relating factors in high-risk breast cancer patients treated with adjuvant standard or high-dose chemotherapy: a longitudinal study. AB - PURPOSE: Determine whether standard or high-dose chemotherapy leads to changes in fatigue, hemoglobin (Hb), mental health, muscle and joint pain, and menopausal status from pre- to post-treatment and to evaluate whether fatigue is associated with these factors in disease-free breast cancer patients. PATIENTS AND METHODS: Eight hundred eighty-five patients were randomly assigned between two chemotherapy regimens both followed by radiotherapy and tamoxifen. Fatigue was assessed using vitality scale (score < or = 46 defined as fatigue), poor mental health using mental health scale (score < or = 56 defined as poor mental health) both of Short-Form 36, muscle and joint pain with Rotterdam Symptom Checklist, and Hb levels were assessed before and 1, 2, and 3 years after chemotherapy. RESULTS: Fatigue was reported in 20% of 430 assessable patients (202 standard dose, 228 high-dose) with at least a 3-year follow-up, without change over time or difference between treatment arms. Mean Hb levels were lower following high dose chemotherapy. Only 5% of patients experienced fatigue and anemia. Mental health score was the strongest fatigue predictor at all assessment moments. Menopausal status had no effect on fatigue. Linear mixed effect models showed that the higher the Hb level (P = .0006) and mental health score (P < .0001), the less fatigue was experienced. Joint (P < .0001) and muscle pain (P = .0283) were associated with more fatigue. CONCLUSION: In 3 years after treatment, no significant differences in fatigue were found between standard and high-dose chemotherapy. Fatigue did not change over time. The strongest fatigue predictor was poor mental health. PMID- 16219927 TI - Not working 3 years after breast cancer: predictors in a population-based study. AB - PURPOSE: Little is known about factors increasing likelihood of not working among breast cancer survivors compared with women in the general population. PATIENTS AND METHODS: A population-based retrospective cohort study was conducted in Quebec, Canada, based on the consecutive series of working women aged younger than 60 years when first treated for breast cancer (identified through the Quebec Tumor Registry), and on a group of randomly selected similar women, living in Quebec, who were working at the time of survivors' diagnoses, but who were without cancer (identified through provincial health care files). Data came from a telephone interview, 3 years after diagnosis for 646 survivors (73% of those eligible) or during a similar period for 890 comparison women (51%). RESULTS: Slightly more survivors were not working 3 years after diagnosis compared with women never diagnosed with cancer (21% and 15%, respectively). Older age (for survivors and comparison women, relative risk [RR] = 4.62, P < .0001 and RR = 4.98, P < .0001, respectively) and union membership (RR = 1.88, P = .0003 and RR = 1.40, P = .06, respectively) increased the likelihood of not working at the end of follow-up. In addition, income less than 20,000 dollars compared with > or = 50,000 dollars was associated with not working only among survivors (RR = 3.18; P = .0008). Adjuvant treatments did not predict work cessation, but any new cancer event during follow-up did (RR = 2.14; P < .0001). CONCLUSION: Although reassuring that adjuvant treatments did not appear to play a role in survivors' not working, other aspects of the cancer experience might nonetheless have influenced the decision to reduce work effort after breast cancer. PMID- 16219928 TI - Mediastinoscopy: an endangered species? PMID- 16219929 TI - Prevalence and causes of fatigue after cancer treatment: the next generation of research. PMID- 16219930 TI - Surgical management of BRCA1 and BRCA2 carriers: bitter choices slightly sweetened. PMID- 16219931 TI - Variations in prostate cancer patterns of care: is it the quality of care or the quality of the data? PMID- 16219932 TI - Non-small-cell lung cancer vaccine therapy: a concise review. AB - Lung cancer is the leading cause of cancer deaths in the United States and throughout the world; globally, there are more than 1.1 million deaths each year. Treatment modalities currently employed are significantly limited; 50% of patients experience disease recurrence after surgery, and less than a quarter of patients respond to systemic chemotherapy. These statistics have fueled the search for a safer, more effective treatment modality. Despite significant advances in our understanding of the molecular basis of cancer immunology, many obstacles remain. However, encouraging clinical results in patients immunized with autologous tumor cell vaccines expressing granulocyte macrophage colony stimulating factor strongly advocate further investigation of immunotherapy in non-small-cell lung cancer (NSCLC). Further studies are needed to demonstrate whether these novel therapies can potentially complement or even replace current therapeutic approaches. We present a review of the various vaccine-based strategies employed to target and treat NSCLC. PMID- 16219933 TI - Galectin-1: a link between tumor hypoxia and tumor immune privilege. AB - PURPOSE: To identify a 15-KDa novel hypoxia-induced secreted protein in head and neck squamous cell carcinomas (HNSCC) and to determine its role in malignant progression. METHODS: We used surface-enhanced laser desorption ionization time of-flight mass spectrometry (SELDI-TOF-MS) and tandem MS to identify a novel hypoxia-induced secreted protein in FaDu cells. We used immunoblots, real-time polymerase chain reaction (PCR), and enzyme-linked immunoabsorbent assay to confirm the hypoxic induction of this secreted protein as galectin-1 in cell lines and xenografts. We stained tumor tissues from 101 HNSCC patients for galectin-1, CA IX (carbonic anhydrase IX, a hypoxia marker) and CD3 (a T-cell marker). Expression of these markers was correlated to each other and to treatment outcomes. RESULTS: SELDI-TOF studies yielded a hypoxia-induced peak at 15 kDa that proved to be galectin-1 by MS analysis. Immunoblots and PCR studies confirmed increased galectin-1 expression by hypoxia in several cancer cell lines. Plasma levels of galectin-1 were higher in tumor-bearing severe combined immunodeficiency (SCID) mice breathing 10% O2 compared with mice breathing room air. In HNSCC patients, there was a significant correlation between galectin-1 and CA IX staining (P = .01) and a strong inverse correlation between galectin-1 and CD3 staining (P = .01). Expression of galectin-1 and CD3 were significant predictors for overall survival on multivariate analysis. CONCLUSION: Galectin-1 is a novel hypoxia-regulated protein and a prognostic marker in HNSCC. This study presents a new mechanism on how hypoxia can affect the malignant progression and therapeutic response of solid tumors by regulating the secretion of proteins that modulate immune privilege. PMID- 16219934 TI - Macrophage and mast-cell invasion of tumor cell islets confers a marked survival advantage in non-small-cell lung cancer. AB - PURPOSE: The role played by the innate immune system in determining survival from non-small-cell lung cancer (NSCLC) is unclear. The aim of this study was to investigate the prognostic significance of macrophage and mast-cell infiltration in NSCLC. METHODS: We used immunohistochemistry to identify tryptase+ mast cells and CD68+ macrophages in the tumor stroma and tumor islets in 175 patients with surgically resected NSCLC. RESULTS: 5-year survival was 52.9% in patients with an islet macrophage density greater than the median versus 7.7% when less than the median (P < .0001). In the same groups, respectively, median survival was 2,244 versus 334 days (P < .0001). Patients with a high islet macrophage density but incomplete resection survived markedly longer than patients with a low islet macrophage density but complete resection. CONCLUSION: The tumor islet CD68+ macrophage density is a powerful independent predictor of survival from surgically resected NSCLC. The biologic explanation for this and its implications for the use of adjunctive treatment requires further study. PMID- 16219935 TI - Endoscopic ultrasound-guided fine-needle aspiration in the diagnosis and staging of lung cancer and its impact on surgical staging. AB - PURPOSE: The diagnosis and staging of lung cancer critically depends on surgical procedures. Endoscopic ultrasound (EUS) -guided fine-needle aspiration (FNA) is an accurate, safe, and minimally invasive technique for the analysis of mediastinal lymph nodes (LNs) and can additionally detect tumor invasion (T4) in patients with centrally located tumors. The goal of this study was to assess to what extent EUS-FNA could prevent surgical interventions. PATIENTS AND METHODS: Two hundred forty two consecutive patients with suspected (n = 142) or proven (n = 100) lung cancer and enlarged (> 1 cm) mediastinal LNs at chest computed tomography were scheduled for mediastinoscopy/tomy (94%) or exploratory thoracotomy (6%). Before surgery, all patients underwent EUS-FNA. If EUS-FNA established LN metastases, tumor invasion, or small-cell lung cancer (SCLC), scheduled surgical interventions were cancelled. Surgical-pathologic verification occurred when EUS-FNA did not demonstrate advanced disease. Cancelled surgical interventions because of EUS findings was the primary end point. RESULTS: EUS-FNA prevented 70% of scheduled surgical procedures because of the demonstration of LN metastases in non-small-cell lung cancer (52%), tumor invasion (T4) (4%), tumor invasion and LN metastases (5%), SCLC (8%), or benign diagnoses (1%). Sensitivity, specificity, and accuracy for EUS in mediastinal analysis were 91%, 100% and 93%, respectively. No complications were recorded. CONCLUSION: EUS-FNA qualifies as the initial staging procedure of choice for patients with (suspected) lung cancer and enlarged mediastinal LNs. Implementation of EUS-FNA in staging algorithms for lung cancer might reduce the number of surgical staging procedures considerably. PMID- 16219936 TI - Effect of short-term hormone replacement therapy on breast cancer risk reduction after bilateral prophylactic oophorectomy in BRCA1 and BRCA2 mutation carriers: the PROSE Study Group. AB - PURPOSE: Bilateral prophylactic oophorectomy (BPO) is widely used for cancer risk reduction in women with BRCA1/2 mutations. Many premenopausal women choose to take hormone replacement therapy (HRT) after undergoing BPO to abrogate immediate symptoms of surgically-induced menopause. Thus, we evaluated whether the breast cancer risk reduction conferred by BPO in BRCA1/2 mutation carriers is altered by use of post-BPO HRT. METHODS: We identified a prospective cohort of 462 women with disease-associated germline BRCA1/2 mutations at 13 medical centers to evaluate breast cancer risk after BPO with and without HRT. We determined the incidence of breast cancer in 155 women who had undergone BPO and in 307 women who had not undergone BPO on whom we had complete information on HRT use. Postoperative follow-up was 3.6 years. RESULTS: Consistent with previous reports, BPO was significantly associated with breast cancer risk reduction overall (hazard ratio [HR] = 0.40; 95%CI, 0.18 to 0.92). Using mutation carriers without BPO or HRT as the referent group, HRT of any type after BPO did not significantly alter the reduction in breast cancer risk associated with BPO (HR = 0.37; 95% CI, 0.14 to 0.96). CONCLUSION: Short-term HRT use does not negate the protective effect of BPO on subsequent breast cancer risk in BRCA1/2 mutation carriers. PMID- 16219937 TI - Accuracy of helical computed tomography and [18F] fluorodeoxyglucose positron emission tomography for identifying lymph node mediastinal metastases in potentially resectable non-small-cell lung cancer. AB - PURPOSE: Computed tomography (CT) and [18F] Fluorodeoxyglucose positron emission tomography (FDG-PET) are considered suitable methods for the noninvasive staging of the mediastinum. Our study was intended to estimate the efficacy of contrast enhanced helical CT (hCT) and FDG-PET, alone and combined, in the diagnosis of lymph node mediastinal metastases. METHODS: This study was a prospective and blind comparison of the efficacy of hCT and FDG-PET with two alternative reference standards, mediastinoscopy, and mediastinoscopy plus thoracotomy plus a 6-month follow-up to diagnose lymph node mediastinal metastases in 132 consecutive patients with potentially resectable non-small-cell lung cancer (NSCLC). The metastatic disease was assessed histopathologically. Further clinical information was obtained postoperatively after a median follow-up of 42 months. RESULTS: The prevalence of cN2,3 is 0.28. For hCT the sensitivity and specificity are 0.86 (95% CI, 0.70 to 0.93) and 0.67 (95% CI, 0.56 to 0.75), for PET 0.94 (95% CI, 0.81 to 0.98) and 0.59 (95% CI, 0.49 to 0.68), and for hCT and PET combined in-parallel 0.97 (95% CI, 0.84 to 0.99) and 0.44 (95% CI, 0.34 to 0.53), which translate into a negative predicted probability of 0.98 (95% CI, 0.88 to 1.00). The crude diagnostic odds ratio of PET in the total sample studied is 13.1, in the subgroup hCT+ 11.04 (3.0 to 40 0.1), and in the hCT- 3.5 (0.5 to 21.5). Similar results were obtained for hCT stratified by PET. CONCLUSION: hCT and PET perform similarly in the mediastinal staging of NSCLC, both tests are conditionally dependent and provide complementary information, and their diagnostic value mainly resides on the negative results. PMID- 16219938 TI - Absolute quantification of proteins by LCMSE: a virtue of parallel MS acquisition. AB - Relative quantification methods have dominated the quantitative proteomics field. There is a need, however, to conduct absolute quantification studies to accurately model and understand the complex molecular biology that results in proteome variability among biological samples. A new method of absolute quantification of proteins is described. This method is based on the discovery of an unexpected relationship between MS signal response and protein concentration: the average MS signal response for the three most intense tryptic peptides per mole of protein is constant within a coefficient of variation of less than +/ 10%. Given an internal standard, this relationship is used to calculate a universal signal response factor. The universal signal response factor (counts/mol) was shown to be the same for all proteins tested in this study. A controlled set of six exogenous proteins of varying concentrations was studied in the absence and presence of human serum. The absolute quantity of the standard proteins was determined with a relative error of less than +/-15%. The average MS signal responses of the three most intense peptides from each protein were plotted against their calculated protein concentrations, and this plot resulted in a linear relationship with an R(2) value of 0.9939. The analyses were applied to determine the absolute concentration of 11 common serum proteins, and these concentrations were then compared with known values available in the literature. Additionally within an unfractionated Escherichia coli lysate, a subset of identified proteins known to exist as functional complexes was studied. The calculated absolute quantities were used to accurately determine their stoichiometry. PMID- 16219939 TI - Are pain location and physical examinations useful in locating a tear site of the rotator cuff? AB - BACKGROUND: Pain is the most common symptom of patients with rotator cuff tendinopathy, but little is known about the relationship between the site of pain and the site of cuff pathologic lesions. Also, accuracies of physical examinations used to locate a tear by assessing the muscle strength seem to be affected by the threshold for muscle weakness, but no studies have been reported regarding the efficacies of physical examinations in reference to their threshold. HYPOTHESIS: Pain location is useful in locating a tear site. Efficacies of physical examinations to evaluate the function of the cuff muscles depend on the threshold for muscle weakness. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The authors retrospectively reviewed the clinical charts of 160 shoulders of 149 patients (mean age, 53 years) with either rotator cuff tears (140 shoulders) or cuff tendinitis (20 shoulders). The location of pain was recorded on a standardized form with 6 different areas. The diagnostic accuracies of the following tests were assessed with various thresholds for muscle weakness: supraspinatus test, the external rotation strength test, and the lift-off test. RESULTS: Lateral and anterior portions of the shoulder were the most common sites of pain regardless of existence of tear or tear location. The supraspinatus test was most accurate when it was assessed to have positive results with the muscle strength less than manual muscle testing grade 5, whereas the lift-off test was most accurate with a threshold less than grade 3. The external rotation strength test was most accurate with a threshold of less than grade 4+. CONCLUSION: The authors conclude that pain location is not useful in locating the site of a tear, whereas the physical examinations aiming to locate the tear site are clinically useful when assessed to have positive results with appropriate threshold for muscle weakness. PMID- 16219940 TI - Risk factors for noncontact ankle sprains in high school athletes: the role of hip strength and balance ability. AB - BACKGROUND: Ankle sprains are among the most common sports injuries. HYPOTHESIS: Poor balance as measured on a balance board and weakness in hip abduction strength are associated with an increased risk of noncontact ankle sprains in high school athletes. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: One hundred sixty-nine high school athletes (101 male athletes, 68 female athletes) from football, men's basketball, men's soccer, women's gymnastics, women's basketball, and women's soccer were observed for 2 years. Balance in single-limb stance on an instrumented tilt board and hip flexion, abduction, and adduction strength (handheld dynamometer) were assessed in the preseason. Body mass, height, generalized ligamentous laxity, previous ankle sprains, and ankle tape or brace use were also documented. RESULTS: There were 20 noncontact inversion ankle sprains. Balance ability (P = .72), hip abduction strength (P = .66), hip adduction strength (P = .41), and hip flexion strength (P = .87) were not significant risk factors for ankle sprains. The incidence of grade II and grade III sprains was higher in athletes with a history of a previous ankle sprain (1.12 vs 0.26 per 1000 exposures, P < .05). A higher body mass index in male athletes was associated with increased risk (P < .05). The combination of a previous injury and being overweight further increased risk (P < .01). CONCLUSION: Balance as measured on a balance board and hip strength were not significant indicators for noncontact ankle sprains. The apparent high injury risk associated with the combination of a history of a previous ankle sprain and being overweight in male athletes warrants further examination. PMID- 16219941 TI - The International Knee Documentation Committee Subjective Knee Evaluation Form: normative data. AB - BACKGROUND: The International Knee Documentation Committee Subjective Knee Evaluation Form may be used to measure symptoms, function, and sports activity for people with a variety of knee disorders, including ligamentous and meniscal injuries, osteoarthritis, and patellofemoral dysfunction. To date, normative data have not been established for this valid, reliable, and responsive outcomes instrument. PURPOSE: To provide clinicians and researchers with normative data to facilitate the interpretation of results on the International Knee Documentation Committee Subjective Knee Evaluation Form. STUDY DESIGN: Cross-sectional survey. METHODS: The Subjective Knee Evaluation Form was mailed to 600 people in each of 8 age/gender categories (18-24 years, 25-34 years, 35-50 years, and 51-65 years for both male subjects and female subjects). Participants were drawn from a panel of 550 000 households (1 300 000 subjects) representative of noninstitutionalized persons in the United States and were matched to data from the United States Census Bureau on geographical region, market size, income, and household size. RESULTS: Complete data were available for 5246 knees. Twenty-eight percent of respondents reported an injury, weakness, or other problem with one or both knees. Normative data were determined for respondents as a whole and for the subset of respondents with no history of knee problems. Mean scores were determined for men aged 18 to 24 years (89 +/- 18), 25 to 34 years (89 +/- 16), 35 to 50 years (85 +/- 19), and 51 to 55 years (77 +/- 23); mean scores were also determined for women aged 18 to 24 years (86 +/- 19), 25 to 34 years (86 +/- 19), 35 to 50 years (80 +/- 23), and 51 to 65 years (71 +/- 26). Scores were higher for the subset of respondents with no history of current or prior knee problems. CONCLUSION: Scores on the International Knee Documentation Committee Subjective Knee Evaluation Form vary by age, gender, and history of knee problems. The normative data collected in this article will allow clinicians to interpret how patients with knee injuries are functioning relative to their age- and gender matched peers and will enable researchers to determine the clinical outcomes of treatment. PMID- 16219942 TI - Elbow range of motion in professional baseball pitchers. AB - BACKGROUND: Physicians involved with the care of baseball players have noted elbow range of motion changes in pitchers. Objective data regarding the extent of these changes have rarely been documented. HYPOTHESIS: Dominant and nondominant elbow range of motion differences are common in baseball pitchers, and these differences are related to player age, amount and length of time professionally pitched, and history of surgical procedures on the dominant extremity. STUDY DESIGN: Cross-sectional study; Level of evidence, 4. METHODS: Thirty-three professional pitchers were evaluated for elbow range of motion during spring training preseason physical examination. Dominant and nondominant elbow range of motion including flexion, extension, supination, and pronation were measured with a goniometer. Range of motion measures from the dominant and nondominant sides were compared. Baseball records were reviewed for arm dominance, age, years of professional pitching, professional innings pitched, and history of elbow surgery. These factors were evaluated for their possible association with range of motion for each side and the difference between sides. RESULTS: Statistically significant differences between dominant and nondominant sides were noted for elbow extension (dominant decreased 7.9 degrees +/- 7.4 degrees, P < .0001), flexion (dominant decreased 5.5 degrees +/- 7.8 degrees, P = .0003), and total flexion-extension arc (dominant decreased 13.3 degrees +/- 13.7 degrees, P < .0001). No significant difference between sides was found for the supination or pronation measures. No correlation was noted for age, pitching history, surgery, or arm dominance and the motion differences. CONCLUSION: Professional pitchers demonstrate elbow flexion and extension differences between dominant and nondominant elbows. No correlation was found between motion differences and age, pitching history, surgery, or arm dominance. PMID- 16219943 TI - The use of a single osteochondral autograft plug in the treatment of a large osteochondral lesion in the femoral condyle: an experimental study in sheep. AB - BACKGROUND: The use of osteochondral autograft plugs can be restricted because of limited amount of donor material. HYPOTHESIS: A small osteochondral autograft plug placed in the center of a large defect in a sheep femoral condyle will yield results superior to either an untreated or a bone-grafted defect. STUDY DESIGN: Controlled laboratory study. METHODS: Twelve adult sheep underwent bilateral hindlimb surgery. On 1 limb, a 6-mm circular osteochondral autograft plug was placed in the center of a 10-mm circular defect in the medial femoral condyle. The gap between the plug and the condyle was filled with bone graft. On the contralateral side, the defect was either left untreated or filled with bone graft (control specimens). Animals were studied at 6 and 12 months under gross examination, high-resolution radiography, and histologic evaluation. RESULTS: At 6 months, 4 of 6 plugs healed and showed good maintenance of the joint surface and cartilage viability in the plugs. One plug fractured and resorbed, and 1 plug settled but healed. At 1 year, all 5 plugs healed, 1 having settled slightly (1 animal died earlier). The plug specimens showed better maintenance of the condyle contour at both times, and the central plug had hyaline-appearing cartilage. The control specimens were more irregular, had a fibrocartilage fill, and appeared flatter, although no gross cavitation or collapse was indicated. Composite cartilage scores on histologic evaluation were significantly higher for the plug specimens after 6 months (P = .02) and 1 year (P = .036) compared with controls. CONCLUSION: At 6 months and 1 year, a 6-mm osteochondral plug placed in a 10-mm defect better preserved the articular surface and contour of the condyle compared to untreated or bone-grafted defects. CLINICAL RELEVANCE: Osteochondral autograft plugs may be able to treat larger articular lesions without complete fill of the defect. PMID- 16219944 TI - An evaluation of the provocative tests for superior labral anterior posterior lesions. AB - BACKGROUND: Although our understanding of superior labral anterior posterior lesions has grown, the physical diagnosis remains imperfect. STUDY DESIGN: Cohort study (diagnostic); Level of evidence, 2. PURPOSE: To determine the most effective provocative maneuver with which to diagnose superior labral anterior posterior lesions. METHODS: A series of 132 consecutive patients scheduled to undergo diagnostic shoulder arthroscopy were examined preoperatively over a 6 month period, and the final diagnosis in each case was made arthroscopically. The following assessments were included: active compression (O'Brien), anterior slide, pain provocation, crank, Jobe relocation, Hawkins, Neer, Speed, and Yergason tests. RESULTS: The most sensitive diagnostic tools for type II superior labral lesions were the active compression, Hawkins, Speed, Neer, and Jobe relocation tests. When type I and type II lesions were combined, the results were similar. However, none of the sensitive tests were specific for either type I or type II lesions. CONCLUSIONS: The authors' results contradict the current literature regarding provocative testing for both stable and unstable superior labral lesions. There is no single maneuver that can accurately diagnose superior labral anterior posterior lesions; arthroscopy remains the standard by which to diagnose such lesions. PMID- 16219945 TI - Pectoralis major tears: correlation of magnetic resonance imaging and treatment strategies. AB - BACKGROUND: Controversy exists in the diagnosis and treatment of pectoralis major tear patterns. HYPOTHESIS: Magnetic resonance imaging is useful in determining the grade and the location of pectoralis major tears and in guiding toward an appropriate treatment plan. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Between January 1998 and December 2002, 27 patients were treated for pectoralis major tears. All patients were evaluated by history and physical examination. A clinical impression was formed regarding the location and grade of injury, and a treatment plan was established. All patients underwent magnetic resonance imaging, and images were reviewed by an orthopaedic surgeon and a board certified musculoskeletal radiologist, resulting in a final treatment plan. The mean follow-up was 12 months. All patients were evaluated by questionnaire and clinical examination to assess pain, function, deformity, and overall satisfaction. RESULTS: In 19 of 27 patients, the clinical impression and magnetic resonance imaging agreed regarding the location and the grade of the injury. A total of 19 patients underwent surgical repair, and 8 patients were treated nonoperatively. The magnetic resonance imaging result changed the treatment plan in 3 patients from operative to nonoperative. CONCLUSION: The clinical impression appeared to overestimate the severity, the location, and the grade of the injury. Magnetic resonance imaging provided a more accurate assessment and, in conjunction with the clinical examination, helped to identify those patients who would benefit most from surgical repair. PMID- 16219946 TI - Human meniscus allografts' in vivo size and motion characteristics: magnetic resonance imaging assessment under weightbearing conditions. AB - BACKGROUND: Although many reports have described clinical outcome of meniscus transplants, their size and motion patterns are not well understood. This study assessed postoperative in vivo size and motion patterns of meniscus transplants under full weightbearing conditions. HYPOTHESIS: The human meniscus transplant has size and motion characteristics similar to the native menisci. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Eight meniscus transplants in 7 knees were studied 15 to 34 months postoperatively. The knees were scanned in an open magnetic resonance imaging 0.5-T superconducting magnet while standing. Single slice sagittal and coronal images were obtained at 0 degrees , 30 degrees , 60 degrees , and 90 degrees of flexion in the operative and contralateral knees. RESULTS: The mean height and width of the anterior and posterior horns of the transplants were similar to those of native menisci. The millimeters of coronal displacement of motion of the midbody were also similar between the transplants and the native menisci. The anterior horn of the native medial menisci moved a mean of 5 mm more (total anterior to posterior translation, P < .05) than did the transplants. The posterior horn of the native medial menisci, and both horns of the native lateral menisci, also tended to move more than the corresponding horns of the transplanted menisci, although this result could not be confirmed statistically, given the number of menisci studied. CONCLUSION: Meniscus transplants had similar size and midbody motion characteristics as the native menisci. The horns of the meniscus transplants tended to show decreased motion compared with the native menisci. The operative techniques and subsequent healing of the graft bone and peripheral attachments provided a stable meniscus construct. PMID- 16219947 TI - Achilles tendon and paratendon microcirculation in midportion and insertional tendinopathy in athletes. AB - BACKGROUND: Neovascularisation can be detected qualitatively by Power Doppler in Achilles tendinopathy. Quantitative data regarding tendon microcirculation have not been established and may be substantial. PURPOSE: To assess the microcirculation of the Achilles tendon and the paratendon in healthy volunteers as well as in athletes with either midportion or insertional tendinopathy. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: In 66 physically active volunteers, parameters of Achilles tendon and paratendon microcirculation, such as tissue oxygen saturation, relative postcapillary venous filling pressures, and microcirculatory blood flow, were determined at rest at 2-mm and 8-mm tissue depths. Forty-one patients never had Achilles pain (25 men, 27 +/- 8 years), 14 patients had insertional pain (7 men, 29 +/- 8 years), and 11 patients had midportion tendinopathy (7 men, 38 +/- 13 years, not significant). RESULTS: Achilles tendon diameter 2 cm and 6 cm proximal to the insertion was increased in symptomatic tendons. Compared with the uninvolved opposite tendon, deep microcirculatory blood flow was significantly elevated at insertional (160 +/- 79 vs 132 +/- 42, P < .05) as well as in midportion tendinopathy (150 +/- 74 vs 119 +/- 34, P < .05). The microcirculation in the uninvolved opposite tendon and the normal athlete controls were not significantly different from each other (132 +/- 42 insertional asymptomatic vs 119 +/- 34 mid-portion vs 120 +/- 48 healthy tendon). Insertional paratendon deep microcirculatory flow was elevated in all groups, whereas tissue oxygen saturation and relative postcapillary venous filling pressures were not significantly different. CONCLUSION: Microcirculatory blood flow is significantly elevated at the point of pain in insertional and midportion tendinopathy. Postcapillary venous filling pressures are increased at both the midportion Achilles tendon and the midportion paratendon, whereas tissue oxygen saturation is not different among the studied groups. We found no evidence of an abnormal microcirculation of the asymptomatic limb in Achilles tendinopathy. PMID- 16219948 TI - Mixing modalities: exploring the options for Graves' ophthalmopathy. PMID- 16219949 TI - Streptococcus suis, Streptococcus mitis, what is the problem? PMID- 16219950 TI - Combined orbital irradiation and systemic steroids compared with systemic steroids alone in the management of moderate-to-severe Graves' ophthalmopathy: a preliminary study. AB - OBJECTIVE: To assess the efficacy and safety of combined orbital irradiation and systemic steroids in the management of moderate-to-severe Graves' ophthalmopathy. DESIGN: Single-blind randomised prospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Sixteen patients with active moderate-to-severe Graves' ophthalmopathy who were randomly assigned to steroid therapy (ST group) or combination therapy of orbital irradiation and systemic steroids (SRT group) between June 2000 and June 2003. MAIN OUTCOME MEASURES: NOSPECS scoring system, total eye score, subjective eye score, and extra-ocular muscle thickness as determined by either computed tomographic or magnetic resonance imaging scans. RESULTS: The study was completed by 15 of 16 patients. Both groups experienced improvement in total eye score, soft tissue swelling, ocular motility, visual acuity, and subjective eye score at 52-week follow-up. Total eye score improved earlier in the SRT group, achieving statistical significance (P<0.05) at as early as 4 weeks of follow-up. Improvement in ocular parameters was greater and led to a significantly greater reduction in total eye score than in the ST group at weeks 16, 24, and 52. Maximum extra-ocular muscle thickness was significantly reduced in the SRT group only. No change was observed in proptosis in either group. No serious adverse effect was observed with the addition of orbital irradiation to steroid therapy. CONCLUSION: A combination of orbital irradiation and systemic steroids is well tolerated and more effective than steroids alone in the treatment of active moderate-to-severe Graves' ophthalmopathy. It achieves greater and more rapid improvement in soft tissue swelling, ocular motility, and visual acuity. PMID- 16219951 TI - Kawasaki disease in Hong Kong, 1994 to 2000. AB - OBJECTIVE: To describe the epidemiology, clinical characteristics, and management of Kawasaki disease in children in Hong Kong. DESIGN: Retrospective survey of medical records from July 1994 to June 1997, and prospective data collection from July 1997 to June 2000. SETTING: Hospitals with a paediatric unit in Hong Kong. PATIENTS: Patients diagnosed with Kawasaki disease between July 1994 and June 2000 in public hospitals in Hong Kong. MAIN OUTCOME MEASURES: Incidence of Kawasaki disease and coronary artery aneurysm rates. RESULTS: A total of 696 cases of Kawasaki disease were reported. There were 435 (62.5%) boys and 261 (37.5%) girls giving a male to female ratio of 1.7:1. The age ranged from 1 month to 15 years 5 months with a median of 1.7 years. Infants (<1 year) constituted the largest group of patients (223, 32.0%) and overall, 638 (91.7%) were younger than 5 years. Skin rash, conjunctivitis, and oral signs were among the principal clinical features present in over 80% of cases. Prominent cervical lymph nodes larger than 1.5 cm were less commonly found (24%). Coronary artery aneurysms or ectasia were present in 15.7% (109/696), 8.5% (59/696), and 5.0% (35/696) of patients at 2, 4, and 8 weeks, respectively. The incidence of Kawasaki disease per 100,000 children under 5 years was significantly higher in the prospective study period than in the retrospective period (39 vs 26, P<0.001). CONCLUSION: The incidence of Kawasaki disease is high in Hong Kong and is 39 per 100,000 children below 5 years of age. The coronary artery aneurysm prevalence is 5%. Intravenous gamma-globulin and high-dose aspirin is the mainstay of treatment. PMID- 16219952 TI - Polycystic ovarian syndrome in Hong Kong Chinese women: patient characteristics and diagnostic criteria. AB - OBJECTIVES: To identify the characteristics of Hong Kong Chinese women with polycystic ovarian syndrome and to compare different diagnostic criteria. DESIGN: Retrospective study. SETTING: Gynae-endocrinology Clinics in the Prince of Wales Hospital, Hong Kong. PATIENTS: Ninety Hong Kong Chinese women with polycystic ovarian syndrome who were diagnosed according to the hospital's criteria. MAIN OUTCOME MEASURES: Prevalence of typical features of polycystic ovarian syndrome, including anovulation and hyperandrogenism (with other endocrine causes excluded), polycystic ovarian features on ultrasonography, luteinising hormone predominance, obesity, and insulin resistance. RESULTS: Almost all (98.9%) patients with polycystic ovarian syndrome presented with anovulation, only 48.9% of them had clinical or biochemical evidence of hyperandrogenism. Typical ultrasound appearances of polycystic ovaries were observed in 86.7% of patients. Luteinising hormone predominance and insulin resistance were demonstrated in 67.8% and 40.7% of the cohort, respectively. Eight-six (95.6%) patients should have also been diagnosed with polycystic ovarian syndrome based on the 2003 Rotterdam new criteria. About 60% of patients who screened positive for insulin resistance had normal fasting serum glucose levels. The same proportion who had full screening for insulin resistance by oral glucose tolerance tests and fasting serum glucose to insulin ratios had discordant results of these two tests. CONCLUSIONS: The 2003 Rotterdam new diagnostic criteria for polycystic ovarian syndrome are generally applicable to the Hong Kong Chinese population. Early detection of insulin resistance in patients with polycystic ovarian syndrome can be ensured by performing an oral glucose tolerance test combined with measurement of fasting serum glucose to insulin ratio. PMID- 16219953 TI - Neuroimpairment, activity limitation, and participation restriction among children with cerebral palsy in Hong Kong. AB - OBJECTIVES: To study children with cerebral palsy in Hong Kong, their neuroimpairment, activity limitation, and participation restriction in society. Parents' opinion on current medical and rehabilitation services was also sought. DESIGN: Systematic survey using questionnaires. SETTING: Four associations in Hong Kong: Child Assessment Service, Hong Kong Association for Parents of Children with Physical Disabilities, Association of Parents of the Severely Mentally Handicapped, and Hong Kong Physically Handicapped and Able-Bodied Association. PARTICIPANTS: Parents of children with cerebral palsy. MAIN OUTCOME MEASURES: Neuroimpairment, activity limitation, and participation restriction. RESULTS: Information from 181 children with cerebral palsy was analysed. Among them, 56% were boys. The mean age was 7 years 6 months (standard deviation, 3 years 11 months). The most common diagnostic type was spastic cerebral palsy. Co morbidities in children with cerebral palsy were common. Limitation in daily activities including mobility and self-care tasks was considerable and this posed great stress to parents when taking care of their children. Children's participation in both social and leisure activities was regarded as a low priority. A high percentage (70%) of parents reported difficulty in travelling. The reasons involved problems in transportation, building access (entry and exit), and attitudes of the general public. These environmental factors restricted the social participation of the children and their families. Over 75% of parents were satisfied with the current medical and rehabilitation services. CONCLUSIONS: Children with cerebral palsy have multiple and complex needs. The findings of this study may serve as a reference for parents, service providers, and policy makers to work in partnership to achieve a more comprehensive health care service for children with cerebral palsy and to facilitate better integration into the community. PMID- 16219955 TI - Surgical management of substernal goitre: local experience. AB - OBJECTIVES: To examine the presentation, workup, and surgical complications of substernal goitre. DESIGN: Retrospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Twenty-four mostly elderly patients (mean age, 60.1 years) who underwent thyroidectomy for substernal goitres between 2000 and 2003 (substernal goitres were defined as those having either a caudal mass transgressing the fourth thoracic vertebra or having more than 50% of their overall mass residing within the thorax). MAIN OUTCOME MEASURES: Symptoms, histopathological diagnoses, morbidities, and complications. RESULTS: Dyspnoea was the most common symptom (n=8, 33%). Three (12.5%) patients presented with acute airway obstruction; however, 13 (54.2%) were asymptomatic apart from the presence of cervical masses. Computed tomographic scans were performed on all but two patients. Malignancy was present in 12.6% of patients, or 16.8% if occult papillary carcinoma is included. Partial or full sternotomies were performed in two (8.3%) patients. Complications included recurrent laryngeal nerve injury (n=1, 2.7% of nerves at risk), transient hypoparathyroidism (n=2, 13.3% of patients at risk), haematoma (n=1, 2.7%), pneumonia (n=1, 2.7%), and wound infection (n=1, 2.7%). There was no operative mortality or permanent hypoparathyroidism. The complication rate was significantly lower in the asymptomatic patients (P=0.033 by Fisher's exact test); clinicopathological parameters were otherwise statistically comparable between the two groups. CONCLUSIONS: There is rarely any mortality in thyroidectomy for substernal goitre, and the morbidity is also very low, especially in asymptomatic patients. In the absence of contra-indications, substernal goitre should be treated with early surgery rather than having it run the risk of acute airway distress or cancer. PMID- 16219954 TI - Curative radiotherapy for early cancers of the lip, buccal mucosa, and nose--a simple interstitial brachytherapy technique employing angiocatheters as carriers for Iridium-192 wire implants. AB - OBJECTIVES: To evaluate treatment outcomes following interstitial brachytherapy for cancers of the lip, buccal mucosa, or nose. DESIGN: Retrospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: A cohort of 13 patients treated uniformly by a simple interstitial brachytherapy technique employing plastic angiocatheters as carriers for Iridium-192 wires: all but one patient had T1 or T2 tumours and all but one had N0 disease. MAIN OUTCOME MEASURES: Local and loco regional control rates. RESULTS: Six of the 13 patients received external radiotherapy prior to interstitial brachytherapy. A median brachytherapy dose of 70 Gy was delivered to those treated with brachytherapy alone, while 35 Gy was delivered after a median external radiotherapy dose of 50 Gy to those receiving combined treatment. The 3-year actuarial local control rate was 75%. No significant late complications were observed. CONCLUSIONS: Employing a simple brachytherapy technique using angiocatheters and Iridium-192 wires, in conjunction with external radiotherapy when appropriate, produces good outcomes for patients with early lip, nasal vestibule, and buccal mucosa cancers. PMID- 16219956 TI - Portal vein embolisation prior to extended right-sided hepatic resection. AB - OBJECTIVES: To determine whether preoperative portal vein embolisation improves the operative outcome of patients undergoing extended right-sided hepatic resection for hepatobiliary malignancy. DESIGN: Prospective non-randomised study. SETTING: University teaching hospital, Hong Kong. PATIENTS: Ninety-two patients underwent extended right-sided hepatic resection for hepatobiliary malignancy during a 45-month period (January 2000 to September 2003). Among them, 15 (16%) underwent portal vein embolisation via a percutaneous ipsilateral approach (n=9) or through the ileocolic vein with a mini-laparotomy (n=6). The remaining 77 (84%) patients underwent hepatic resection without portal vein embolisation. MAIN OUTCOME MEASURES: Operative morbidity and mortality. RESULTS: Patients undergoing portal vein embolisation were older (69 years vs 55 years; P=0.009), and had significantly worse preoperative renal function (creatinine, 96 micromol/L vs 86 micromol/L; P=0.039) and liver function (bilirubin, 23 micromol/L vs 12 micromol/L; P<0.001). Portal vein embolisation resulted in an increase in the future liver remnant of 9% (interquartile range, 7-13%) of the estimated standard liver volume. The operating time for patients receiving portal vein embolisation was significantly longer (medium, 660 min vs 420 min; P<0.001) with more complicated surgery performed in terms of concomitant caudate lobectomy and hepaticojejunostomy. There was no hospital mortality in patients who underwent portal vein embolisation whereas five without the treatment died (P=0.587). The operative morbidity of patients who underwent portal vein embolisation and those who did not was 20% and 30%, respectively (P=0.543). CONCLUSIONS: In older patients who have worse preoperative liver and renal functions, portal vein embolisation enhances the possibility to perform extended right-sided hepatic resection for hepatobiliary malignancies with potentially lower operative mortality and morbidity. PMID- 16219957 TI - Child behaviour and parenting stress in Hong Kong families. AB - OBJECTIVES: To examine parent perception of child behaviour problems and parenting stress in Hong Kong, and to assess the extent to which they are related to socio-demographic factors and the availability of social support. DESIGN: Cross-sectional survey by using a questionnaire. SETTING: Maternal and Child Health Centres, Hong Kong. PARTICIPANTS: Parents of children aged 4 years who were registered with Maternal and Child Health Centres and were living in Hong Kong between September 2002 and February 2003. MAIN OUTCOME MEASURES: Child behaviour problems and parenting stress. RESULTS: A total of 1009 questionnaires were returned giving a participation rate of 67.0%. About one tenth of parents were experiencing difficulties with their children's behaviour. Parenting stress and children's behaviour problems were associated with presence or absence of social support. Parenting stress was also associated with household income. CONCLUSIONS: The prevalence of child behaviour problems in Hong Kong is comparable with international figures. Intervention programmes should be targeted at parents who experience difficulties with their children's behaviour and parenting. PMID- 16219958 TI - Influenza vaccination: options and issues. AB - Currently available vaccines have similar efficacy if they are matched to the most prevalent circulating strains. They also have comparable adverse effect profiles. The choice of a specific preparation of vaccine therefore requires consideration of cost, purity of the vaccine preparation in terms of the amount of egg protein and endotoxin, allergy to different constituents of the vaccine, reactogenicity profiles, as well as the preferred route of administration. Intradermal injection of the vaccine appears to be a viable alternative to the traditional intramuscular administration with the additional benefit of requiring a smaller volume of vaccine. Despite the documented benefits in various community and institutional settings, influenza vaccination has been underutilised by most target groups. A major obstacle to broader coverage of vaccination is the perceived ineffectiveness of the vaccine and the relatively benign nature of the illness in most patients. Uptake of vaccine among target populations, especially health care workers, needs to be improved through a concerted effort between frontline clinicians and health authorities. PMID- 16219959 TI - The clinical outcome of 137 rape victims in Hong Kong. AB - From 1 August 2001 to 31 July 2004, 137 patients were referred from the Association Concerning Sexual Violence Against Women to the Accident and Emergency Department at the Kwong Wah Hospital for alleged rape. Approximately half of the patients presented within 3 days of the alleged assault. Fifty-one patients were prescribed emergency contraception: one patient remained pregnant despite treatment and was referred with a further six patients to the Gynaecology Department for termination of pregnancy. Thirty-two patients received hepatitis B immunoglobulin injection. One patient had a positive result for rapid plasma reagin 3 months following the assault and was referred to the Social Hygiene Clinic. All tests for antibody to human immunodeficiency virus were negative. Antimicrobial therapy was prescribed for women who had an endocervical and/or high vaginal swab positive for Chlamydia trachomatis (n=9), Trichomonas vaginalis (n=1), and gonococcus (n=1). PMID- 16219960 TI - Diarrhoea and rash in a retired farmer. AB - An 83-year-old retired farmer developed invasive strongyloidiasis after using potent topical corticosteroids. Initial colonoscopy features mimicked ulcerative colitis, and the clinical course was complicated by bacterial septicaemia with Serratia and Enterococcus. He was treated successfully with oral ivermectin and antibiotics. PMID- 16219961 TI - Herpetic shoulder paresis in a Chinese elderly patient. AB - A patient with left shoulder girdle weakness secondary to herpetic myotomal paresis is reported. Needle electromyography revealed denervational discharge from the left supraspinatus, deltoid, and brachioradialis muscles, compatible with a radiculopathy that was relevant to his myotomes affected by zoster infection. The patient was managed with range-of-movement and strengthening exercises as well as pain relief for post-herpetic neuralgia. Further studies are required to determine whether antiviral treatment can limit the extent of motor deficit and hasten recovery. Zoster paresis should be one of the differential diagnoses of girdle muscle weakness. PMID- 16219962 TI - A case of human brucellosis in Hong Kong. AB - Brucellosis is an infectious disease of humans and animals caused by Brucella species. We report on a 34-year-old housewife who presented with recurrent headache, fever, and malaise. Blood cultures yielded slow-growing gram-negative coccobacilli that were later identified as Brucella melitensis. The patient recalled handling goat placenta in China. She was prescribed a 6-week course of doxycycline and rifampicin. Laboratory staff who had been exposed to the isolate remained asymptomatic. The epidemiology, diagnosis, and treatment of brucellosis are discussed. PMID- 16219963 TI - Fever, pulmonary haemorrhage, and acute renal failure in a young girl. AB - Leptospirosis is rarely reported in children. It is easily treated with antibiotics but can be fatal if treatment is delayed. A 12-year-old girl was diagnosed with Weil syndrome, a severe form of leptospirosis, 10 days after returning from Mainland China. She presented with fever, hypotension, jaundice, and acute renal failure. She had used well water for bathing and swum in freshwater. PMID- 16219964 TI - A case of Streptococcus mitis endocarditis successfully treated by linezolid. AB - We report the successful treatment of infective endocarditis caused by Streptococcus mitis with linezolid in a patient with pre-existing valvular heart disease. The patient had multiple allergies to conventional antibiotics. Linezolid may provide an oral alternative in the treatment of infective endocarditis in patients with adverse drug reactions to traditional antibiotic regimens. PMID- 16219965 TI - Clopidogrel-induced hepatotoxicity after percutaneous coronary stenting. AB - Clopidogrel, an adenosine diphosphate receptor blocker, is widely used as an adjunctive antiplatelet therapy in acute coronary syndrome and percutaneous coronary stenting. The occurrence of hepatotoxicity is rare. We describe the occurrence of symptomatic liver disease in a 74-year-old man 5 weeks following commencement of therapy with clopidogrel. The reported cases of clopidogrel induced hepatotoxicity are reviewed and the clinical significance of this event are discussed. PMID- 16219966 TI - Acute abdominal pain in a patient with a history of ocular melanoma. PMID- 16219967 TI - The contractual model of the patient-physician relationship and the demise of medical professionalism. PMID- 16219968 TI - Jehovah's Witnesses and surgery. PMID- 16219969 TI - Anti-inflammatory action of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers and the prevention of diabetes mellitus in obese patients with hypertension. PMID- 16219970 TI - [Is Icelandic natural environment as clean as we like to think?]. PMID- 16219971 TI - [The subspecialties of medicine. Reflexions on the status of the subspecialties in an Icelandic university hospital]. PMID- 16219972 TI - [Swimmer's itch in Landmannalaugar, Iceland]. AB - Swimmer's itch (SI) or human cercarial dermatitis is caused by free-swimming larvae of bird parasites of the family Schistosomatidae (Trematoda) which have penetrated thorough the skin. Sometimes, mainly during first infections, the larvae do not cause any symptoms but if trapped by the immune system of the host each larva causes a maculopapular eruption. So far, five bird schistosome species have been reported in Iceland. Cercariae are shed by the freshwater snail Radix peregra but adults live in anseriform birds; one nasal Trichobilharzia species occurs in the nasal cavity of mallards, two visceral species have been found in veins of whooper swans and a visceral species has been found in greylag goose and in mallards, respectively. Experiments have shown that developing schistosomulae are able to survive for days or even weeks in mammals. Long term pathologic effects on the host are unknown. During the second half of August 2003 thousands of bathers got SI in a slowly streaming brook with geothermally heated groundwater in Landmannalaugar, the most frequently visited area in the interior of Iceland. The number of cercariae in the water and SI cases decreased until October but still in December and in late winter 2004 SI cases were reported. In August 2004 SI again started in the area but the density of cercariae in the water seemed to be less than in the previous year. The prevalence of snails shedding Trichobilharzia cercariae on the bathing site never exceeded 1%. The rapid increase of cercariae in the water by the middle of August in 2003 and 2004 were caused by a breeding mallard female and its ducklings which were raised on the bathing site during summer. All the ducklings had nasal- and visceral Trichobilharzia infections which they must have acquired soon after hatching. Three weeks later the adult worms could have started egg-laying. Consequently, emerging miracidia infected the snails which finally started shedding the cercariae by middle of August in 2003 and 2004. In future years this rapid increase of cercariae could be avoided if ducklings are not allowed to have access to the bathing site and the adjacent brook. Retrospective enquiries suggest that some visitors have occasionally got SI after bathing in the brook in preceding decades. Low number of papules, however, indicates a low density of cercariae in the water. PMID- 16219973 TI - [Knee arthroplasties performed at Akureyri University Hospital in the years 1983 2003. Results with emphasis on revision and complication rates]. AB - OBJECTIVE: Osteoarthrosis (OA) is a growing medical problem in western societies and the cost of the treatment has grown accordingly in the last years. Patients with OA often need to be operated on with arthroplasties and one important outcome measure for this type of surgery is the revision rate. The purpose of this study was to assess the results of knee arthroplasties performed at Akureyri University Hospital during 1983-2003, with special emphasis on revision rates, infections and other complications. MATERIAL AND METHODS: Information was gathered from journals of the 457 patients who underwent knee arthroplasties during 1983-2003. Information about the operation and hospital stay was recorded as well as if patients later became the subject of revision. CRR (cumulative revision rate) uses survival statistics to estimate the risk of revision after primary operation and was calculated for patients with OA operated on with knee arthroplasty. Statistical informations were calculated in Microsoft Excel. Kaplan Mayer analysis was used to calculate the CRR and that was done in SPSS 11.5. RESULTS: 560 primary operations were performed during the period, 515 total knee arthroplasties and 45 unicompartmental. 200 operations were performed on males and 360 on females. Mean ages for males was 70.8 years and for females 69.4 years. Revision rates varied depending on the type of implant. Twelve unicompartmental and 28 total knee arthroplasties became subject of revision. The PCA unicompartmental prosthesis most frequently needed revision, or in over 50% of cases. The CRR for the AGC total knee prosthesis was the lowest or around 3% at seven years, including revisions due to infections. Revisions due to infections were three in the period or 0.6% of all the total knee arthroplasties. Complications that substantially increase the risk of revision and/or are life threatening were recorded in 1.8% of the operations at the time of discharge. Only one patient had pulmonary embolism (0.2%) and two patients (0.4%) had deep venous thrombosis. CONCLUSION: Our high revision rate for the PCA implant is consistent with what has been seen in other studies. This prosthesis was found to have mechanical problems and was withdrawn from the market. Our revision rate for the AGC implant as well as the rate of infections are low and the results are quite comparable to what has been found in Sweden by the Swedish Knee Arthroplasty Registry. The results of knee arthroplasties performed at Akureyri University Hospital, regarding revision rates, infections and complications, are fully comparable to other known results internationally. PMID- 16219974 TI - [The correlation between toe- and ankle pressure, clinical symptoms and angiography in patients with leg ischemia]. AB - OBJECTIVE: The severity of leg ischemia is usually evaluated by measuring ankle pressure. This is a simple measurement but can be misleading in patients with severe sclerosis in ankle arteries in which case toe pressure is believed to be more reliable. The purpose of this study was to compare toe pressure with ankle pressure, clinical symptoms and angiography and thus evaluate the usefulness of toe pressure in the assessment of leg ischemia. MATERIAL AND METHODS: In total of 58 legs from 30 patients that came to Landspitali University Hospital because of leg ischemia we measured toe- and ankle pressure and assessed the clinical stage of leg ischemia. All patients also had an angiography of their leg arteries. RESULTS: There was a correlation between pressure measurements and the clinical stage of leg ischemia and also between pressure measurements and results from angiography of leg arteries. There was not a significant difference between the correlation for toe- and ankle pressure respectively. In two cases it was impossible to measure ankle pressure but in both cases it was possible to measure toe pressure. CONCLUSIONS: Toe- and ankle pressure measurements are equally useful in the evaluation of leg ischemia. Ankle pressure is a more simple measurement than toe pressure but can be impossible to perform when the ankle arteries are very sclerotic. When that is the case toe pressure can often be measured instead. PMID- 16219975 TI - The role of mitochondrial rRNAs and nanos protein in germline formation in Drosophila embryos. AB - Germ cells, represented by male sperm and female eggs, are specialized cells that transmit genetic material from one generation to the next during sexual reproduction. The mechanism by which multicellular organisms achieve the proper separation of germ cells and somatic cells is one of the longest standing issues in developmental biology. In many animal groups, a specialized portion of the egg cytoplasm, or germ plasm, is inherited by the cell lineage that gives rise to the germ cells (germline). Germ plasm contains maternal factors that are sufficient for germline formation. In the fruit fly, Drosophila, germ plasm is referred to as polar plasm and is distinguished histologically by the presence of polar granules, which act as a repository for the maternal factors required for germline formation. Molecular screens have so far identified several of these factors that are enriched in the polar plasm. This article focuses on the molecular functions of two such factors in Drosophila, mitochondrial ribosomal RNAs and Nanos protein, which are required for the formation and differentiation of the germline progenitors, respectively. PMID- 16219976 TI - Functional involvement of Xenopus homologue of ADF/cofilin phosphatase, slingshot (XSSH), in the gastrulation movement. AB - ADF/cofilin is a phosphorylation-regulated protein essential for actin filament dynamics in cells. Here, we cloned two cDNAs encoding Xenopus ADF/cofilin (XAC) specific phosphatase, slingshot (XSSH), one of which contains an extra 15 nucleotides in a coding sequence of the other, possibly generated by alternative splicing. Whole mount in situ hybridization showed XSSH transcripts in the blastopore lip and sensorial ectoderm at stage 11, and subsequently localized to developing brain, branchial arches, developing retina, otic vesicle, cement gland, and spinal chord in neurula to tailbud embryos. Immunostaining of animal vegetal sections of gastrula embryos demonstrated that both XAC and XSSH proteins are predominant in ectodermal and involuting mesodermal cells. Microinjection of either a wild type (thus induces overexpression) or a phosphatase-defective mutant (functions as dominantly negative form) resulted in defects in gastrulation, and often generated the spina bifida phenotype with reduced head structures. Interestingly, the ratio of phosphorylated XAC to dephosphorylated XAC markedly increased from the early gastrula stage (stage 10.5), although the amount of XSSH protein markedly increased from this stage. These results suggest that gastrulation movement requires ADF/cofilin activity through dynamic regulation of its phosphorylation state. PMID- 16219977 TI - Involvement of slingshot in the Rho-mediated dephosphorylation of ADF/cofilin during Xenopus cleavage. AB - ADF/cofilin is a key regulator for actin dynamics during cytokinesis. Its activity is suppressed by phosphorylation and reactivated by dephosphorylation. Little is known, however, about regulatory mechanisms of ADF/cofilin function during formation of contractile ring actin filaments. Using Xenopus cycling extracts, we found that ADF/cofilin was dephosphorylated at prophase and telophase. In addition, constitutively active Rho GTPase induced dephosphorylation of ADF/cofilin in the egg extracts. This dephosphorylation was inhibited by Na(3)VO (4) but not by other conventional phosphatase-inhibitors. We cloned a Xenopus homologue of Slingshot phosphatase (XSSH), originally identified in Drosophila and human as an ADF/cofilin phosphatase, and raised antibody specific for the catalytic domain of XSSH. This inhibitory antibody significantly suppressed the Rho-induced dephosphorylation of ADF/cofilin in extracts, suggesting that the dephosphorylation at telophase is dependent on XSSH. XSSH bound to actin filaments with a dissociation constant of 0.4 microM, and the ADF/cofilin phosphatase activity was increased in the presence of F-actin. When latrunculin A, a G-actin-sequestering drug, was added to extracts, both Rho induced actin polymerization and dephosphorylation of ADF/cofilin were markedly inhibited. Jasplakinolide, an actin-stabilizing drug, alone induced actin polymerization in the extracts and lead to dephosphorylation of ADF/cofilin. These results suggest that Rho-induced dephosphorylation of ADF/cofilin is dependent on the XSSH activation that is caused by increase in the amount of F actin induced by Rho signaling. XSSH colocalized with both actin filaments and ADF/cofilin in the actin patches formed on the surface of the early cleavage furrow. Injection of inhibitory antibody blocked cleavage of blastomeres. Thus, XSSH may reorganize actin filaments through dephosphorylation and reactivation of ADF/cofilin at early stage of contractile ring formation. PMID- 16219978 TI - Induction of female-to-male sex reversal by high temperature treatment in Medaka, Oryzias latipes. AB - Medaka, Oryzias latipes, has a firm XX-XY sex-determining system with the sex determining gene, DMY, on the Y chromosome. However, previous studies have suggested that high water temperature might affect sex determination in Medaka. In the present study, the influence of high water temperature on sex reversal was examined. Fertilized eggs of two inbred strains of Medaka were developed at high water temperature (32 degrees C) until hatching. The hatched fry were kept at normal water temperatures (27 degrees C) until adulthood, and the phenotypic and genotypic sex was examined. As a result, 24% (N=105) and 50% (N=36) of XX fish developed a male phenotype in the Hd-rR and HNI inbred strains, respectively. These XX sex-reversed males had a normal testis and were fully fertile. On the other hand, all XY fish were male in the both strains. These results demonstrate that high water temperatures can induce XX sex reversal and that elevated water temperatures during the embryonic stage is a simple and useful method for getting XX males in Medaka. PMID- 16219979 TI - Territorial behavior of a green hairstreak Chrysozephyrus smaragdinus (Lepidoptera: Lycaenidae): site tenacity and wars of attrition. AB - Males of Chrysozephyrus smaragdinus were active from late morning to late afternoon, during which they showed territorial behavior, perhaps for mating. The territorial male stayed in a particular area and occasionally flew around it, referred to hereafter as the inspection area. When other male intruded into this area, the territorial male rushed to him. Then, they engaged in a circling flight regarded as a "war of attrition". During this flight, the two males sometimes strayed far away from the territory. After the circling flight, the resident returned to his territory in almost all cases (98%). Despite such intrusions, many residents defended their territory for several successive days. This suggests strongly the "effect of prior residence". We recorded the circling flights with a high-speed video camera, and confirmed that the male that ceased the circling flight first was the loser. This finding gave some validity to consider circling flight as wars of attrition. In a few cases, the territorial male mated with a female that came to the territory. These once mated males held the territory no longer, suggesting that mating experience should restrict the next mating opportunity in this species. PMID- 16219980 TI - Distribution of a novel protein AgK114 expression in the normal tissues of adult mice: dual expression of AgK114 and growth hormone in anterior pituitary cells. AB - The novel antigen K114 (AgK114) has been previously identified in normal hamster skin, and its expression has been up-regulated accompanying tissue damages of the skin, although there is no information on its biological functions. To determine the physiological role of AgK114, we prepared anti-mouse AgK114 monoclonal antibody and studied its tissue distribution in healthy adult mice by immunocytochemistry. A widespread and unique expression of AgK114 peptide was found in the selected organs of various systems (hair follicle cells and sebaceous gland of skin, ciliated epithelial cells of trachea and bronchial tube, striated portion of submandibular gland, distal convoluted tubule cells of kidney, ciliated epithelial cells of oviduct, medulla of adrenal gland and anterior lobe of pituitary gland). Interestingly, dual expression of AgK114 peptide and growth hormone in somatotrophs was found in anterior lobe of pituitary gland by double immunocytochemistry. AgK114 peptide was expressed widely in many regionally well-defined cellular systems in various peripheral tissues, suggesting that AgK114 peptide may have some roles of physiological functions in these organs. The data from our current study have provided a rationale for further studies of functional roles of AgK114 peptide in a variety of organs or tissues under physiological conditions. PMID- 16219981 TI - Interleukin-18 (IL-18) mRNA expression and localization of IL-18 mRNA-expressing cells in the mouse uterus. AB - Interleukin-18 (IL-18) belongs to the interleukin-1 family and was identified as an interferon-gamma inducing factor. We investigated IL-18 mRNA-expressing cells in the mouse uterus. By RNase protection assay, IL-18 mRNA and alpha subunit of IL-18 receptor mRNA were detected in the uterus. In the uterus, IL-18 mRNA levels increased during sexual maturation. In situ hybridization analysis demonstrated IL-18 mRNA-expressing cells in the mouse uterus of different ages. At 21 days of age, IL-18 mRNA-expressing cells were detected in the luminal epithelial cells and stromal cells although the IL-18 mRNA signal was weak. At 42 days of age, IL 18 mRNA signal was mainly detected in the stromal cells located near the myometrium, and in some of the luminal and glandular epithelial cells. In the uterus of 63-day-old adult mice, a strong hybridization signal for IL-18 mRNA was detected at estrus, but was weak at diestrus. IL-18 mRNA was mainly detected in the glandular epithelial cells and stromal cells. The effect of estradiol-17beta (E(2)) on IL-18 mRNA-expressing cells in the uterus was examined in ovariectomized mice. In oil-treated mice IL-18 mRNA signal was localized in luminal epithelial cells and stromal cells, while in E(2)-treated mice IL-18 mRNA signal was localized in stromal cells alone. These results suggest that the mouse uterus has an IL-18 system, and IL-18 exerts a physiological role within the uterus in a paracrine manner, and that IL-18 gene expression is regulated by estrogen. PMID- 16219983 TI - Development of non-radioisotopic immunoassay systems for measuring flounder IGF I. AB - A time-resolved fluoroimmunoassay system (TR-FIA) for measuring flounder insulin like growth factor-I (IGF-I) was developed using biotinylated flounder IGF-I, anti-fish IGF-I antiserum and europium-avidin conjugate. The detection limit per well was <5 pg/well corresponding to <0.5 ng/ml in a basic procedure for sample of 10 microl/well and to <0.08 ng/ml in a procedure modified for high volume samples (up to 70 microl/well). Specificity of the assay was validated using various IGF-Is and insulins. All IGFs except seabream IGF-I showed very low or no crossreactivity. Binding inhibition curves for flounder and seabream IGF-Is were completely identical to each other. Intra- and interassay variations ranged from coefficients of variations of 3.9% to 7.2%. Recovery tests using barfin flounder plasma varied from 82.7 to 101.6% in the added range from 20 to 160 ng/ml. This assay system was applied for measuring total plasma IGF-I in barfin flounder injected porcine growth hormone (GH). A group injected with GH at the dose of 0.05 IU/gBW showed a significant increase of total plasma IGF-I compared with those of albumin-injected (control) and initial groups. In addition, I was able to substitute time-resolved fluorometric detection in this assay system with enzymatic fluorometric detection (FIA). Binding inhibition curve for flounder IGF I in this substituted assay system showed equal performance with that of the TR FIA system. Correlation of IGF-I levels between TR-FIA and FIA was high (r(2)=0.957) in plasma samples from barfin flounders in various physiological conditions. Thus, the present study shows precision and efficiency of two non radioisotopic immunoassay systems for measuring flounder IGF-I. PMID- 16219982 TI - Organ-specific and age-dependent expression of insulin-like growth factor-I (IGF I) mRNA variants: IGF-IA and IB mRNAs in the mouse. AB - Insulin-like growth factor-I (IGF-I) gene generates several IGF-I mRNA variants by alternative splicing. Two promoters are present in mouse IGF-I gene. Each promoter encodes two IGF-I mRNA variants (IGF-IA and IGF-IB mRNAs). Variants differ by the presence (IGF-IB) or absence (IGF-IA) of a 52-bp insert in the E domain-coding region. Functional differences among IGF-I mRNAs, and regulatory mechanisms for alternative splicing of IGF-I mRNA are not yet known. We analyzed the expression of mouse IGF-IA and IGF-IB mRNAs using SYBR Green real-time RT PCR. In the liver, IGF-I mRNA expression increased from 10 days of age to 45 days. In the uterus and ovary, IGF-I mRNA expression increased from 21 days of age, and then decreased at 45 days. In the kidney, IGF-I mRNA expression decreased from 10 days of age. IGF-IA mRNA levels were higher than IGF-IB mRNA levels in all organs examined. Estradiol-17beta (E2) treatment in ovariectomized mice increased uterine IGF-IA and IGF-IB mRNA levels from 3 hr after injection, and highest levels for both mRNAs were detected at 6 hr, and relative increase was greater for IGF-IB mRNA than for IGF-IA mRNA. These results suggest that expression of IGF-I mRNA variants is regulated in organ-specific and age dependent manners, and estrogen is involved in the change of IGF-I mRNA variant expression. PMID- 16219984 TI - Molecular phylogeny of Megaloceros giganteus--the giant deer or just a giant red deer? AB - Two fragments of mitochondrial DNA (mtDNA) of the cytochrome b gene (137 bp and 167 bp) were successfully isolated and sequenced from antlers and bones of five specimens of the Giant Deer (Megaloceros giganteus) to examine the phylogenetic position of Megaloceros giganteus within the family Cervidae. This is the first report on ancient DNA (aDNA) sequences from Megaloceros giganteus. A phylogenetic analysis based on parameter-rich models describes the evolutionary relationships between five individuals of fossil Megaloceros giganteus and 37 individuals of 11 extant species of the family Cervidae. The results support a "Cervus-Megaloceros" clade. The phylogenetic positions of sympatric Megaloceros and Cervus elaphus specimens in particular indicate either that the Megaloceros mtDNA gene pool did not evolve for a substantial time period as an entity distinct from Cervus elaphus until its extinction, or that Megaloceros contributed mtDNA to Cervus elaphus or vice versa. The results of this study allow the conclusion that the European Megaloceros giganteus is more related to its modern regional counterparts of the species of Cervus elaphus than recent claims have suggested. PMID- 16219985 TI - Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9. AB - Sry (sex-determining region on the Y chromosome) is required for testicular differentiation in mammals. In addition to Sry, other genes such as WT1, Fgf9, Dax1, Dmrt1 and Sox9 are widely accepted to be involved in the sex determination in vertebrates. However, the roles of these genes during sex determination still remain unclear in amphibians. This study was undertaken to examine the expression of WT1 and Fgf9 in the developing gonad of amphibians. We first isolated the WT1 cDNA from the frog Rana rugosa. Like WT1 in mice, R. rugosa WT1 showed 2 isoforms; i.e., one had an additional 3 amino acids, KTS, included between the third and fourth zinc fingers. However, 17 amino acids in exon 5 of mammalian WT1 could not be found in R. rugosa WT1, which is also the case in turtle and chicken. The mRNA of both isoforms (+KTS, -KTS) was detected in the lung, kidney and testis, but not in the ovary and muscle of adult frogs. The 2 isoforms were expressed first in the embryos at stage 23. Thereafter, the expressions remained constant in the gonad attached to mesonephros of both sexes during sex determination. We next isolated the R. rugosa Fgf9 cDNA encoding 208 amino acids. The amino acid sequence of Fgf9 had similarity greater than 92% with chicken, mouse and human Fgf9s, suggesting that Fgf9 is highly conserved among vertebrate classes. Fgf9 was expressed in the ovary of an adult frog strongly, but in the lung weakly. In contrast, the Fgf9 mRNA was hardly detected in the kidney, testis and muscle. Moreover, Fgf9 did not show a sexually dimorphic expression pattern during sex determination in R. rugosa. The results, taken together, suggest that both WT1 and Fgf9 are expressed in the indifferent gonad prior to sex determination without any difference in the expression between males and females. Thus, it seems unlikely that they are a key factor to initiate the divergence leading to testicular or ovarian differentiation in R. rugosa. PMID- 16219986 TI - Checklist of African Tischeriidae (Insecta: Lepidoptera) with a redescription of the formerly neglected Tischeria urticicolella from Equatorial Africa. AB - Tischeria urticicolella (Ghesquiere, 1940)--a poorly studied species from the Democratic Republic of Congo and formerly known as a member of the Gracillariidae -is redescribed and reported as belonging to the Tischeriidae for the first time. The external features and male genitalia are figured and described. A checklist and distribution map for all 18 Tischeriidae species currently recorded from Africa (including the Canary Islands) are given. PMID- 16219987 TI - German vegan study: diet, life-style factors, and cardiovascular risk profile. AB - BACKGROUND/AIM: Evaluation of cardiovascular risk profile in 154 German vegans. METHODS: Cross-sectional study, Germany. Study instruments: 2 FFQ, 2 questionnaires, analyses of fasting venous blood samples. RESULTS: The total study population had a low BMI (mean: 22.3 kg/m(2)), a moderate blood pressure (mean: 120/75 mm Hg), an extremely low consumption of alcohol (mean: 0.77 g/day) and 96.8% were nonsmokers. Moderate physical activity (PAL) was reported by nearly 50%, whereas 22.7% declared to have a high PAL (>3 h/week). Median triacylglycerol (TG) was 0.81 mmol/l, total cholesterol (TC) was 4.33 mmol/l, HDL was 1.34 mmol/l. The mean TC/HDL-ratio was 3.3. Lipoprotein(a) (Lp(a)) was 8.13 mg/dl, concentrations of >30 mg/dl were prevalent in 25% of the participants. In general, status of folate and pyridoxine were sufficient, while 49.7% showed cobalamin concentrations <150 pmol/l. Plasma homocysteine levels were slightly elevated (median: 12.5 micromol/l). Cobalamin concentration and duration of vegan nutrition were the main determinants of homocysteine in the total study population. CONCLUSION: Although TC and LDL concentrations were favorable, low HDL and elevated homocysteine and Lp(a) concentrations were unfavorable. Overall, these results confirm the notion that a vegan diet is deficient in vitamin B(12), which may have an unfavorable effect on CHD risk. PMID- 16219988 TI - Nutrition rehabilitation of HIV-infected and HIV-negative undernourished children utilizing spirulina. AB - The objective of this study was to assess the impact of an alimentary integrator composed of spirulina (Spirulina platensis; SP), produced at the Centre Medical St Camille of Ouagadougou, Burkina Faso, on the nutritional status of undernourished HIV-infected and HIV-negative children. We compared two groups of children: 84 were HIV-infected and 86 were HIV-negative. The duration of the study was 8 weeks. Anthropometric and haematological parameters allowed us to appreciate both the nutritional and biological effect of SP supplement to traditional meals. Rehabilitation with SP shows on average a weight gain of 15 and 25 g/day in HIV-infected and HIV-negative children, respectively. The level of anaemia decreased during the study in all children, but recuperation was less efficient among HIV-infected children. In fact 81.8% of HIV-negative undernourished children recuperated as opposed to 63.6% of HIV-infected children (Z: 1.70 (95% CI -0.366, -0.002, p = 0.088)). Our results confirm that SP is a good food supplement for undernourished children. In particular, rehabilitation with SP also seems to correct anaemia and weight loss in HIV-infected children, and even more quickly in HIV-negative undernourished children. PMID- 16219989 TI - Resting energy expenditure, cardiovascular risk factors and insulin resistance in obese patients. AB - OBJECTIVE: The aim of our study was to determine whether energy expenditure modified by increasing body mass over the wide range of body mass index (BMI) was related to insulin resistance, cardiovascular risk factors and dietary intakes. SUBJECTS AND METHODS: A population of 87 obese non-diabetic outpatients was analyzed prospectively. Indirect calorimetry, tetrapolar electrical bioimpedance, serial assessment of nutritional intake using written 3-day food records and biochemical analyses were performed. RESULTS: The mean age was 45.1 +/- 16.7 years and the mean BMI was 35.2 +/- 5.2. Indirect calorimetry showed a resting metabolic rate (RMR) of 1,732.2 +/- 406.6 kcal/day and oxygen consumption of 266.2 +/- 63.3 ml/min. RMR corrected by fat-free mass was 36.8 +/- 14.1 kcal/day/kg. Serial assessment of nutritional intake using written 3-day food records showed a calorie intake of 1,660 +/- 551.7 cal/day, a carbohydrate intake of 168.63 +/- 76.6 g/day, a fat intake of 72.1 +/- 26.42 g/day and a protein intake of 81.1 +/- 23.3 g/day. Insulin, HOMA, systolic blood pressure, diastolic blood pressure, waist circumference, fat mass, waist-to-hip ratio were higher in the third BMI tertile than in the first and second tertiles. No differences were detected in calorie intake, carbohydrate intake, fat intake, protein intake, drinking and alcoholic habit among BMI tertiles. RMR was similar in the different tertiles and the corrected RMR by fat-free mass was higher in first tertile than in the second and third tertiles (44.2 +/- 20.7 vs. 34.5 +/- 9.1 vs. 33.1 +/- 8.5 kcal/kg/day; p < 0.05). In multivariate analysis with a dependent variable (RMR), the fat-free mass remained in the model (F = 7.8; p < 0.05), with an increase of 10.1 (95% CI 3.6-17.5) kcal/day with each 1 kg of fat-free mass adjusted by age and sex. CONCLUSION: Resting energy expenditure in obese patients is not related to the BMI, insulin resistance and dietary intake. RMR is related to fat-free mass in a multivariant model. PMID- 16219990 TI - A high-cholesterol, n-3 polyunsaturated fatty acid diet causes different responses in rats and hamsters. AB - This study was designed to investigate the response to a high-cholesterol, n-3 polyunsaturated fatty acid (PUFA) or n-6 PUFA diet in rats and hamsters. Animals were fed n-3 or n-6 PUFA with a cholesterol-free diet, or with a diet enriched with cholesterol (0.5%, w/w) for 2 weeks. In rats and hamsters fed a cholesterol free diet, plasma cholesterol, triglycerides and very-low-density lipoprotein (VLDL)-triglyceride levels in n-3 PUFA group were significantly lower than those in n-6 PUFA group. In contrast, when diets were supplemented with 0.5% cholesterol, the plasma cholesterol- and triglyceride-lowering effect of dietary n-3 PUFA disappeared. In hamsters fed with the atherogenic diet (0.5% dietary cholesterol) for 2 weeks, n-3 PUFA induced hypercholesterolemia more than n-6 PUFA, the increase being in the VLDL and low-density lipoprotein (LDL) fractions. Our data thus indicate that elevation of VLDL- and LDL-cholesterol in hamsters by n-3 PUFA, compared with n-6 PUFA, is dependent on 0.5% dietary cholesterol supplementation. In rats, on the other hand, dietary n-3 PUFA did not induce hypercholesterolemia more than n-6 PUFA when 0.5% cholesterol was supplemented. Although the effects of n-3 PUFA on plasma cholesterol, triglycerides and VLDL triglycerides were similar in hamsters and rats, the interactive effects of n-3 PUFA and cholesterol on plasma and lipoprotein cholesterol levels differed in the two species. It was also found that plasma triglycerides, cholesterol and lipoprotein cholesterol levels in hamsters are higher than in rats in the presence and absence of dietary cholesterol. In addition, cholesterol feeding induces hypertriglyceridemia and hypercholesterolemia only in hamsters. Moreover, liver triglyceride concentrations increased in rats fed a cholesterol-rich diet and hepatic triglyceride levels of the n-3 PUFA-fed rats were significantly lower than those in the n-6 PUFA-fed rats in the presence and absence of dietary cholesterol. However, triglycerides did not accumulate in the liver in hamsters fed a cholesterol-rich diet and hepatic triglyceride levels of the n-3 PUFA-fed hamsters were not significantly different from those in the n-6 PUFA-fed hamsters in the presence and absence of dietary cholesterol. Therefore, these studies confirm marked species differences in response to the interactive effects of dietary n-3 PUFA and cholesterol. PMID- 16219991 TI - Sketches of otohistory. Part 11: Ototoxicity: drug-induced hearing loss. PMID- 16219992 TI - Clinical application of dichotic multiple-stimulus auditory steady-state responses in high-risk newborns and young children. AB - Experience with dichotic multiple-stimulus auditory steady-state responses (ASSRs) in clinical practice is described. ASSR thresholds were assessed in a sample of 60 high-risk newborns and young children between birth and 4 years of age. Amplitudes and signal-to-noise ratios (SNRs) of the ASSR were compared between normal-hearing infants and adults. Age-related changes within a group of infants younger than 3 months of age were investigated. A comparison was made between ASSR, the click-evoked auditory brainstem response and behavioral hearing thresholds in infants with a wide range of hearing threshold levels. Mean ASSR thresholds for normal-hearing infants at an average corrected age of 12 days were 42 +/- 10, 35 +/- 10, 32 +/- 10 and 36 +/- 9 dB SPL for 0.5, 1, 2 and 4 kHz, respectively. Compared to adults, these thresholds were elevated by on average 11 dB and SNRs were 1.7 times smaller. However, based on ASSRs, reasonably accurate estimations could be made of behavioral hearing thresholds obtained at a later age (median delay of 7 months). The predicted thresholds were in 61% of the cases within 10 dB of the corresponding behavioral thresholds, and in 83% of the cases within 15 dB. In less than 1 h, thresholds at four frequencies per ear could be obtained. The optimal age of testing is between 1 week and 3 months corrected age. The dichotic multiple-stimulus ASSR technique is a valuable extension of the clinical test battery for hearing-impaired children, as a follow-up diagnostic after the neonatal hearing screening. PMID- 16219994 TI - An evoked potential study of the developmental time course of the auditory nerve and brainstem in children using cochlear implants. AB - Central auditory responses to electrical stimulation from a cochlear implant were studied in 75 pre-lingually deafened children and 11 adults. Electrically evoked auditory brainstem response (EABR) latencies significantly decreased with duration of cochlear implant use and were not significantly affected by the age at implant activation. Significant decreases in early latency waves and interwaves occurred within the first 1-2 months of implant use, whereas longer term changes (6-12 months) were found for eV and eIII-eV, which measure activity in the more rostral brainstem. Comparisons to acoustically evoked auditory brainstem response (ABR) in children with normal hearing suggested shorter interwave EABR latencies, reflecting either distinct neural generators or increased neural synchrony, but similar rates of change in the later latency eV and eIII-eV with time in sound. In sum, normal-like development of the rostral auditory brainstem is promoted by cochlear implant use in children of a wide range of ages. PMID- 16219993 TI - Improved music perception with explicit pitch coding in cochlear implants. AB - Music perception and appraisal is very poor in cochlear implant (CI) subjects partly because (musical) pitch is inadequately transmitted by the current clinically used sound processors. A new sound processing scheme (F0mod) was designed to optimize pitch perception, and its performance for music and pitch perception was compared in four different experiments to that of the current clinically used sound processing scheme (ACE) in six Nucleus CI24 subjects. In the F0mod scheme, slowly varying channel envelopes are explicitly modulated sinusoidally at the fundamental frequency (F0) of the input signal, with 100% modulation depth and in phase across channels to maximize temporal envelope pitch cues. The results of the four experiments show that: (1) F0 discrimination of single-formant stimuli was not significantly different for the two schemes, (2) F0 discrimination of musical notes of five instruments was three times better with the F0mod scheme for F0 up to 250 Hz, (3) melody recognition of familiar Flemish songs (with all rhythm cues removed) was improved with the F0mod scheme, and (4) estimates of musical pitch intervals, obtained in a musically trained CI subject, matched more closely the presented intervals with the F0mod scheme. These results indicate that explicit F0 modulation of the channel envelopes improves music perception in CI subjects. PMID- 16219995 TI - Is the western scrub-jay (Aphelocoma californica) really an underdog among food caching corvids when it comes to hippocampal volume and food caching propensity? AB - Food caching has been linked to better performance on spatial memory tasks and enlarged hippocampal volume in both birds and mammals. Within food-caching birds, it has also been predicted that species less reliant on stored food should have inferior spatial memory and a smaller hippocampus compared to species that depend heavily on food caches. Several comparisons suggest that North American corvids have a significantly smaller hippocampus and overall brain volume compared to the Eurasian corvid species and that western scrub-jays (Aphelocoma californica) have a smaller hippocampus compared to the more specialized Clark's nutcracker. Here we present the largest data set of scrub-jay brains and, in contrast to previous reports, show that relative to body mass western scrub-jays have a brain size similar to the largest brain size of Eurasian corvids. The relative hippocampal volume of scrub-jays is also among the largest of all investigated corvids. These findings may not be surprising considering that scrub-jays have been reported to have remarkable cognitive capacities such as episodic-like memory and experience projection. Our data suggest that many previously made assumptions about western scrub-jays as less specialized food hoarders might be an oversimplification and that simple categorization of species into specialized and non-specialized hoarders might not provide useful insights into the evolution of memory and the hippocampus. PMID- 16219996 TI - Cellular localization of AMPA type glutamate receptor subunits in the basal ganglia of pigeons (Columba livia). AB - Corticostriatal and thalamostriatal projections utilize glutamate as a neurotransmitter in mammals and birds. The influence on striatum is mediated, in part, by ionotropic AMPA-type glutamate receptors, which are heteromers composed of GluR1-4 subunits. Although the cellular localization of AMPA-type subunits has been well characterized in mammalian basal ganglia, their localization in avian basal ganglia has not. We thus carried out light microscopic single- and double label and electron microscopic single-label immunohistochemical studies of GluR1 4 distribution and cellular localization in pigeon basal ganglia. Single-label studies showed that the striatal neuropil is rich in GluR1, GluR2, and GluR2/3 immunolabeling, suggesting the localization of GluR1, GluR2 and/or GluR3 to the dendrites and spines of striatal projection neurons. Double-label studies and perikaryal size distribution determined from single-label material indicated that about 25% of enkephalinergic and 25% of substance P-containing striatal projection neuron perikarya contained GluR1, whereas GluR2 was present in about 75% of enkephalinergic neurons and all substance-P -containing neurons. The perikaryal size distribution for GluR2 compared to GluR2/3 suggested that enkephalinergic neurons might more commonly contain GluR3 than do substance P neurons. Parvalbuminergic and calretininergic striatal interneurons were rich in GluR1 and GluR4, a few cholinergic striatal interneurons possessed GluR2, but somatostatinergic striatal interneurons were devoid of all subunits. The projection neurons of globus pallidus all possessed GluR1, GluR2, GluR2/3 and GluR4 immunolabeling. Ultrastructural analysis of striatum revealed that GluR1 was preferentially localized to dendritic spines, whereas GluR2/3 was found in spines, dendrites, and perikarya. GluR2/3-rich spines were generally larger than GluR1 spines and more frequently possessed perforated post-synaptic densities. These results show that the diverse basal ganglia neuron types each display different combinations of AMPA subunit localization that shape their responses to excitatory input. For striatal projection neurons and parvalbuminergic interneurons, the combinations resemble those for the corresponding cell types in mammals, and thus their AMPA responses to glutamate are likely to be similar. PMID- 16219998 TI - Intra- and extrauterine maturation of amplitude-integrated electroencephalographic activity in preterm infants younger than 30 weeks of gestation. AB - OBJECTIVE: To prospectively investigate the longitudinal changes of amplitude integrated electroencephalographic (aEEG) activity in preterm infants <30 weeks gestational age (GA). METHODS: Infants (GA <30 weeks) without evidence of neurological abnormalities had weekly aEEG recordings performed. The relative duration of the three aEEG patterns (discontinuous low voltage, discontinuous high voltage and continuous) was determined and the influence of GA and postnatal age (PNA) on the occurrence of each pattern was assessed. RESULTS: Ninety-eight infants (median GA 26 weeks; range 23-29 weeks) were studied. With higher GA (OR 1.68, 95% CI 1.33-2.13) and PNA (OR 1.91, 95% CI 1.53-2.38), the likelihood for the occurrence of continuous activity increased. The discontinuous low-voltage pattern was less likely to occur with increasing GA (OR 0.68, 95% CI 0.55-0.83) and PNA (OR 0.70, 95% CI 0.61-0.81). CONCLUSION: Maturation of aEEG activity in preterm infants is influenced by both GA and PNA. PMID- 16219997 TI - The eye of the laboratory mouse remains anatomically adapted for natural conditions. AB - Evolutionary effects of domestication have been demonstrated for several body systems, including the eye, and for several vertebrate species, including the mouse. Given the importance of the laboratory mouse to vision science, we wished to determine whether the anatomical and histological features of the eyes of laboratory mice are distinct from those of their naturally adapted, wild counterparts. We measured dimensions and masses of whole eyes and lenses from a wild population plus three inbred strains (C57BL/6J, NZB/BINJ, and DBA/1J) of the house house, Mus musculus, as well as wild and outbred laboratory-domesticated stock of the deer mouse, Peromyscus maniculatus. Histological preparations from these eyes were used to determine outer nuclear layer thickness, linear density of the ganglion cell layer, and for indirect immunofluorescence evaluation of cone opsin expression. For all of these traits, no statistically significant differences were found between any laboratory strain and its wild counterpart. The evolutionary effects of domestication of mice therefore do not include changes to the eye in any variable measured, supporting the continued use of this animal as a model for a naturally adapted visual system. PMID- 16219999 TI - Partial liquid ventilation with low-dose perfluorochemical and high-frequency oscillation improves oxygenation and lung compliance in a rabbit model of surfactant depletion. AB - BACKGROUND: Partial liquid ventilation (PLV) with perfluorochemical (PFC) has been advocated as a new therapy for acute respiratory distress syndrome in both clinical and animal studies, meconium aspiration syndrome, and RDS. PFC is referred to as liquid PEEP because it gets distributed to the most gravity dependent regions of the lung due to its density. High-frequency oscillation (HFO) has been shown to prevent both acute and chronic lung injury in the management of very low birth weight infants with RDS, with gentle ventilation approach. Specifically, HFO with aggressive and adequate lung volume recruitment has been shown to reduce the incidence of chronic lung disease in very low birth weight infants. We hypothesized that PLV along with HFO might be effective in ARDS in an adult rabbit model. OBJECTIVES: To examine the efficiency of low-dose PLV with with HFO on pulmonary gas exchange and lung compliance in a surfactant depleted rabbit model. METHODS: After induction of severe lung injury by repeated saline lung lavage, 19 adult white Japanese rabbits were randomized into two groups that received PLV with HFO (n=9) or HFO gas ventilation (n=10). Physiological and blood gas data were compared between the two groups by analysis of variance. RESULTS: The HFO-PLV group showed improved total lung compliance with maintenance of significantly lower mean airway pressure as compared with the HFO-GAS group so as to keep SpO2>90%. CONCLUSIONS: The addition of a low dose of PFC with HFO was effective in achieving adequate oxygenation, with a reduction in further lung injury in neonates. PMID- 16220000 TI - Angiographic findings of moyamoya vessels in a patient with rheumatoid arthritis. PMID- 16220001 TI - Finding associations in dense genetic maps: a genetic algorithm approach. AB - Large-scale association studies hold promise for discovering the genetic basis of common human disease. These studies will consist of a large number of individuals, as well as large number of genetic markers, such as single nucleotide polymorphisms (SNPs). The potential size of the data and the resulting model space require the development of efficient methodology to unravel associations between phenotypes and SNPs in dense genetic maps. Our approach uses a genetic algorithm (GA) to construct logic trees consisting of Boolean expressions involving strings or blocks of SNPs. These blocks or nodes of the logic trees consist of SNPs in high linkage disequilibrium (LD), that is, SNPs that are highly correlated with each other due to evolutionary processes. At each generation of our GA, a population of logic tree models is modified using selection, cross-over and mutation moves. Logic trees are selected for the next generation using a fitness function based on the marginal likelihood in a Bayesian regression frame-work. Mutation and cross-over moves use LD measures to pro pose changes to the trees, and facilitate the movement through the model space. We demonstrate our method and the flexibility of logic tree structure with variable nodal lengths on simulated data from a coalescent model, as well as data from a candidate gene study of quantitative genetic variation. PMID- 16220002 TI - Relationship between CTLA-4 and CD28 molecule expression on T lymphocytes and stimulating and blocking autoantibodies to the TSH-receptor in children with Graves' disease. AB - The present study was performed to elucidate the relationship between CTLA-4/CD28 molecules and stimulating (TSAb) and blocking (TBAb) antibodies to the TSH receptor (TSH-R) in Graves' disease. CD28 and CD152 (CTLA-4) are glycoprotein molecules which provide a potent costimulatory signal for T-cell activation and proliferation via interactions with their ligands, B7.1/B7.2 molecules, which are present on the surface of antigen-presenting cells. The aim of the study was to estimate the expression of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4, CD152), CD28, B7.1 (CD80), and B7.2 (CD86) molecules on peripheral blood cells in patients with Graves' disease (GD) (n = 55, mean age 15.5 +/- 5.1 years) and nontoxic nodular goiter (NTNG) (n = 55, mean age 15.2 +/- 4.5 years), in comparison with sex and age-matched healthy control subjects (n = 55, mean age 15.2 +/- 3.9 years). The expression of the costimulatory molecules on mononuclear cells was analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. Detection of TSAb and TBAb to the TSH-R using JPO9 CHO cells in unfractionated serum was measured by a highly sensitive commercial radioimmunoassay. When compared with healthy control subjects and euthyroid patients with GD, untreated patients with GD showed a significant increase of CD152+ (p < 0.001, p < 0.001) and CD28+ (p < 0.01, NS) T lymphocytes, respectively. After 6-12 months of methimazole therapy, the percentage of these cells in the peripheral blood of hyperthyroid patients returned to normal values. In addition, patients with GD showed an increase in the percentage of both B7.1 (3.8%) and B7.2 (18.4%) molecules on activated monocytes, compared to patients with NTNG (0.5% p < 0.05, 2.5% p < 0.01, respectively) and healthy control subjects (0.2% p < 0.05, 0.8% p < 0.003, respectively). In patients with untreated GD there was a statistically significant positive correlation between the expression of CTLA-4 on the surface of peripheral blood T cells and the index of TSAb antibodies (R = 0.54, p < 0.001) as well as a negative correlation with TBAb antibody titer (R = -0.58, p < 0.001). However, no such correlations were noted with regard to CD28 and anti-TPO, anti-TG, and TRAb antibodies. We conclude that changes in the expression of costimulatory molecules on the surface of peripheral blood T cells and their significant relationship with the level of antithyroid antibodies indicate an involvement of these molecules in the pathogenesis of GD. PMID- 16220003 TI - Identification of potentially cross-reactive peanut-lupine proteins by computer assisted search for amino acid sequence homology. AB - BACKGROUND: Allergic cross-reactions are an issue of major concern because of implications for public health. The molecular basis of cross-allergy is the similarity of epitopes belonging to proteins of different organisms. Lupine is an emerging cause of food allergy, which has become a 'hot topic' because of recent large-scale introduction into processed foods and frequent cross-reactions with other members of the legume family. However, no lupine allergen has been characterized thus far. Prompted by a recently reported case of peanut-lupine cross-allergy, we wished to identify the possible cross-reactive allergen(s) between the two vegetal species. METHODS: We used computer-aided amino acid sequence comparison, a well-established technique for the study of protein homology, and followed the FAO/WHO guidelines for the identification of potential allergens. We also performed a three-dimensional modeling of the suspected cross reactive proteins to compare their molecular surfaces. RESULTS: We found a highly significant sequence homology and molecular similarity between allergen Ara h 8 of peanut and pathogenesis-related protein PR-10 of white lupine. Another protein of lupine, the beta-conglutin precursor, was found to be significantly homologous to the Ara h 1 allergen of peanut. The molecular surfaces of Ara h 8 and PR-10 were remarkably similar. CONCLUSIONS: Our in silico data allow to predict the allergenicity of PR-10 and beta-conglutin precursor of white lupine according to FAO/WHO guidelines. Amino acid sequence homology also suggests that these proteins could be responsible, at least in part, for some of the allergic cross reactions between peanut and lupine reported in the literature. PMID- 16220004 TI - Dendritic cells in atopic dermatitis: expression of FcepsilonRI on two distinct inflammation-associated subsets. AB - BACKGROUND: Dendritic cells (DCs) represent a major portion within the infiltrate of atopic dermatitis (AD) lesions. As antigen-presenting cells they have the ability to regulate both the quantity and quality of T-cell responses and, thus, are likely to play a key role in the pathogenesis of T-cell-dominated skin diseases such as AD. Thus we sought to identify the DC repertoire occurring in AD patients. METHODS: For this purpose, we phenotypically analyzed various defined DC subsets of AD patients and healthy controls in skin biopsies and peripheral blood by immunofluorescence staining. RESULTS: In AD lesions, two inflammation associated DC subsets with varying expression of costimulatory molecules occurred besides epidermal Langerhans cells (LCs) and dermal myeloid DCs (dmDCs) indigenously residing in normal skin: (1) CD1a+/CD1c+/FcepsilonRI+/IgE+/CD207- myeloid DCs (mDCs) in the epidermis and dermis and (2) CD123+/BDCA 2+/CD45RA+/CD68+ plasmacytoid DCs (pDCs) in the dermis. In the peripheral blood of the patients, these cells exhibited an immature phenotype. Interestingly, we found FcepsilonRI and cell-bound IgE to be expressed not only on myeloid, but also on plasmacytoid DCs from both the skin and peripheral blood of AD patients. CONCLUSIONS: It is tempting to speculate that the disease-regulating role of inflammatory DCs in AD is influenced by both FcepsilonRI occupancy and their degree of maturity. PMID- 16220005 TI - IgE-mediated rat mast cell triggering with tryptic and synthetic peptides of bovine beta-lactoglobulin. AB - BACKGROUND: Immunoglobulin E (IgE) epitopes of beta-lactoglubulin (betaLG) have been identified by ELISA inhibition methods using sera from allergic patients. However, the functional capacity of these epitopes to stimulate mast cells is unknown. It is the goal of the present study to identify bivalent IgE epitopes of betaLG able to trigger target mast cells. METHODS: Peptides were obtained either by purification from tryptic hydrolysates of betaLG or by synthesis. They were examined for their triggering activity in vitro on peritoneal 3H-serotonin labeled rat mast cells passively sensitized with IgE anti-betaLG antibodies. In vivo, rats immunized with betaLG were administered peptides by gavage for intestinal rat mast cell protease II release. RESULTS: Compared with intact betaLG, purified or synthetic tryptic-like betaLG peptides have a sharply decreased allergenicity. Peptide 149-162 retains the highest bivalent IgE epitope mediated triggering capacity. CONCLUSION: A functional bivalent IgE epitope was identified at the C terminal end of betaLG. PMID- 16220006 TI - Exposure to endotoxins during sensitization prevents further endotoxin-induced exacerbation of airway inflammation in a mouse model of allergic asthma. AB - BACKGROUND: We have shown previously that lipopolysaccharides (LPS) inhibited airway inflammation in allergen-sensitized and challenged mice when administered during sensitization, while exacerbating the inflammation when given upon challenge. We have here investigated the effect of LPS administered during both sensitization and challenge on airway inflammation, as well as on the profile of the T-helper (Th) response to allergen. METHODS: Mice were sensitized and challenged with ovalbumin (OVA), in the presence or absence of effective doses of LPS, namely 1 mug during sensitization and 1 ng during challenge. Inflammation was assessed by measuring cell counts and cytokine levels in bronchoalveolar lavage fluid (BALF). The profile of the Th response was determined by quantifying OVA-specific IgE and IgG2a in serum and Th1/Th2 cytokines in the culture medium of splenocytes and in BALF. RESULTS: Allergen-induced airway eosinophilia was increased in mice exposed to LPS during challenge only when compared with controls, whereas it was similarly reduced in animals exposed during sensitization only and during both sensitization and challenge. Mice exposed to LPS during sensitization only or during both sensitization and challenge also displayed a decrease in IgE and an increase in IgG2a, suggesting a switch in the immune response toward the Th1 profile. This was confirmed by quantification of Th1/Th2 cytokines in culture medium of splenocytes and in BALF. CONCLUSIONS: Our data demonstrate that exposure to endotoxins during sensitization prevents allergen-induced airway inflammation, as well as its exacerbation triggered by further exposure to endotoxins during challenge, while switching the immune response to allergen from a Th2 to a Th1 profile. PMID- 16220007 TI - Effects of captopril on cardiac and renal damage, and metabolic alterations in the nitric oxide-deficient hypertensive rat. AB - Chronic inhibition of nitric oxide (NO) synthesis is characterized by increased blood pressure accompanied with both cardiac hypertrophy as well as renal damage. We investigated whether the angiotensin-converting enzyme (ACE) inhibitor captopril can inhibit the cardiac hypertrophy and reverse the renal failure. We tested the influence of captopril on the nitrate-nitrite (NO(x)) in plasma and heart and kidney tissues. Oxidative stress, in terms of glutathione and thiobarbituric acid-reactive substances measured as malondialdehyde, was monitored examining their involvement in the cardioprotective and renoproptective actions. Three groups of Wistar rats were used: untreated group, and rats treated with the NO synthase inhibitor N(w)-nitro-L-arginine methyl ester (L-NAME) and L NAME plus captopril (10 mg/kg/day). Systolic, diastolic and mean blood pressure (BPs, BPd and BPm respectively) was measured weekly in addition to the heart rate using rat-tail plethysmography. After 3 weeks, L-NAME significantly increased BPs, BPd and BPm. Captopril treatment reversed the increments in pressure back to normal values by the fourth week. ACE inhibition by captopril reverted the L-NAME induced hypertrophy and inhibited the enzymatic indices of cardiac damage (glutamate oxaloacetate transaminase and lactate dehydrogenase) back to normal values. Furthermore, the NO synthesis inhibition produced renal damage as indicated by significant increase in creatinine. Captopril ameliorated the raised creatinine to normal. Chronic L-NAME treatment increased serum NO(x) levels but concomitant treatment with captopril was without effect. PMID- 16220008 TI - Antimicrobial activity of a series of 1-alkyl-2-(4-pyridyl)pyridinium bromides against Gram-Positive and Gram-Negative bacteria. AB - OBJECTIVE: To test a series of 1-alkyl-2-(4-pyridyl)pyridinium bromides with alkyl chains containing between 9 and 16 carbons against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli, Stenotrophomonas maltophilia, Acinetobacter baumannii and Pseudomonas aeruginosa) bacteria. MATERIALS AND METHODS: Chemical synthesis was based on the reaction of 2,4' bipyridyl with alkyl bromide. Antimicrobial activity of the bipyridyls was measured by growing bacterial cultures on Mueller-Hinton agar in the presence and absence of inhibitors. RESULTS: The compounds were most active against S. aureus. The most active compounds had alkyl chain lengths of between 11 and 16 carbons. Methicillin-sensitive S. aureus was more susceptible to the inhibitors than methicillin-resistant S. aureus (MRSA). Two subclasses of MRSA existed which differed in their susceptibility to the inhibitors. The susceptibility of MRSA strains to the compounds was increased in the presence of the efflux pump inhibitor reserpine. The activity of the compounds against Gram-negative organisms was increased when the membrane-permeabilizing agent sodium citrate was introduced. Critical micelle concentrations of the compounds were much higher than minimum inhibitory concentrations of the inhibitors. CONCLUSION: The mechanism of action of the compounds may involve perturbing bacterial membranes. The resistance of some MRSA strains to the compounds may be related to efflux pumps. PMID- 16220009 TI - Comparison of polymorphonuclear leukocyte functions in elderly patients and healthy young volunteers. AB - OBJECTIVE: To compare the polymorphonuclear leukocyte (PMN) functions (phagocytosis and intracellular killing activity) of elderly patients with healthy young volunteers. SUBJECTS AND METHODS: Fifty-nine elderly patients who had various diseases (cancer, hypertension and diabetes mellitus, DM) and 10 healthy young volunteers were included in this study. Ficoll-Hypaque gradient centrifugation was used to isolate PMNs from venous blood containing EDTA (0.1 g/ml). Phagocytosis and intracellular killing activity of neutrophils were assayed using a modification of Alexander's method, in which serum opsonins, number of neutrophils and number of microorganisms are standardized in order to detect both increases and decreases in phagocytosis and intracellular killing as well as combined abnormalities of these two functions. The least significant difference test was used to compare the results in the two groups. RESULTS: Phagocytic activity of PMNs from patients with cancer was significantly higher than that of healthy young volunteers (p < 0.05) and elderly patients with hypertension and DM (p < 0.05). There was no statistical difference between the phagocytic activity of PMNs from elderly patients with hypertension and DM and healthy young volunteers (p > 0.05). The intracellular killing activity of PMNs from elderly patients with hypertension, DM and cancer was significantly lower than that of healthy young volunteers (p = 0.001, p < 0.0001, p = 0.003, respectively). CONCLUSIONS: The intracellular killing activity of PMNs from elderly patients was significantly decreased when compared with that of healthy young volunteers. Ageing, chronic diseases and drugs used in the treatment of these elderly patients may be the cause for decreased intracellular killing activity. PMID- 16220010 TI - Renal artery stenosis in patients with peripheral vascular disease in Kuwait. AB - OBJECTIVE: The aim of this study was to identify the incidence of atherosclerotic renal artery stenosis (RAS) in patients with peripheral vascular disease (PVD) and its relation to any known risk factors. SUBJECTS AND METHODS: This prospective study was conducted on 212 patients who were subjected to peripheral angiography for symptoms of PVD over a 3-year period from 1995 to 1998 at the Mubarak Al-Kabeer Hospital, Kuwait. Angiographic evidence of atherosclerotic disease and its severity was recorded in renal, abdominal aorta, iliac, femoral, popliteal and below-knee arteries. In addition, a detailed search of identifiable risk factors was done using history, clinical examination and laboratory studies. RESULTS: The incidence of significant atherosclerotic RAS (more than 50% diameter stenosis) in patients with PVD was 15/212 (7.07%) with no significant difference in ratio between males and females (p = 0.3) compared to that of PVD alone. Patients with common iliac and femoral artery lesions had a high incidence of RAS (93.3 and 86.7%, respectively) with more than 80% probability in RAS patients with involvement of these vessels. There was significant renal impairment (p < 0.005), as assessed by serum creatinine levels, in patients with RAS compared to those who did not have it. There was a high incidence of smoking in patients with RAS (p = 0.02), and smoking was the only risk factor identified in these subjects. CONCLUSIONS: Patients with iliac or femoral atherosclerotic disease have a high probability of associated RAS. Presence of renal impairment in patients with PVD is highly indicative of RAS. Smoking is the only identified risk factor for RAS in association with PVD in our population. PMID- 16220011 TI - Prevalence and severity of malocclusion in adolescent Kuwaitis. AB - OBJECTIVES: Our aim was to evaluate prevalence and severity of malocclusion in an adolescent Kuwaiti population. SUBJECTS AND METHODS: Using a stratified cluster sampling method and excluding non-Kuwaitis, subjects with mesial migration and/or loss of first molars, and subjects with orthodontic treatment experience without availability of initial study models, we examined 1,299 Kuwaitis (674 boys and 625 girls) of mean age 13.2 +/- 0.4 years in a regular well lit classroom. We scored molar and canine relationship, overjet, overbite, anterior and posterior cross bite as well as spacing and irregularity of the incisors using gloves, mirrors, rulers, and spatulas. RESULTS: Fourteen, 15, and 71% met the criteria for almost ideal, mild, and moderate to severe malocclusion, respectively. Of the 86% with malocclusion, occurrence of class 1, half step class 2, full step class 2, half step class 3, and full step class 3 was 57.8, 24.9, 6.3, 8.8, and 2.2% for the first molars, and 36.1, 50.0, 6.2, 5.4, and 2.3% for the canines, respectively. Incisor malalignment was the most prevalent malocclusion trait (73.2%). Overjet > or = 9.5 mm occurred in less than 1.5%, open bite in less than 3.5%, deep bite with gingival contact in less than 2.0%, complete posterior cross bite in less than 1.5%, and complete anterior crossbite in less than 2.0%. Median diastema was present in about 2/3 of the cases with maxillary anterior spacing as opposed to only about 1/3 of those with mandibular spacing. CONCLUSION: Our findings suggest that more than 70% of young adolescent Kuwaitis have moderate to severe malocclusion, with incisor malalignment as the most prevalent malocclusion trait. PMID- 16220013 TI - Spectrum and antibiotic resistance of uropathogens isolated from hospital and community patients with urinary tract infections in two large hospitals in Kuwait. AB - OBJECTIVES: To determine the spectrum of microbial etiology and antibiotic resistance pattern of the uropathogens that cause urinary tract infections in 2 large teaching hospitals in Kuwait over a period of 1 year. MATERIALS AND METHODS: The Vitek identification card system was used to identify the uropathogens. Susceptibility of the isolates against 18 antibiotics was performed by the microbroth dilution method using the Vitek automated system. In addition, gram-positive bacteria were tested in parallel by the disk diffusion technique. RESULTS: The six overall most common isolates were: Escherichia coli, accounting for 47% of isolates in both hospitals, followed by Candida spp. (10.8%), Klebsiella pneumoniae (9.6%), Streptococcus agalactiae (GBS; 9.5%), Enterococcus faecalis (4.2%) and Pseudomonas aeruginosa (4.1%). Amikacin provided the widest coverage amongst all the antibiotics tested followed by ciprofloxacin, gentamicin and piperacillin-tazobactam. For the gram-negatives, high resistance (26-63%) to the beta-lactam antibiotics was noted, especially to ampicillin, amoxicillin clavulanic acid, cephalothin and cefuroxime. Resistance to trimethoprim sulfamethoxazole was also high. None of the enterococci was resistant to the glycopeptides, but 38-60% of the Staphylococcus haemolyticus were resistant to vancomycin or teicoplanin. CONCLUSION: These data show the high level of antimicrobial resistance amongst the uropathogens causing urinary tract infection in the two hospitals studied. PMID- 16220012 TI - Primary thyroid malignancies in Tehran, Iran. AB - OBJECTIVE: The aim of this study was to present a descriptive epidemiology of primary thyroid cancer in Tehran, Iran, using cancer registry data. SUBJECTS AND METHODS: All cases of thyroid cancer registered from 1998 to 2001 in Tehran Metropolitan Area Population-Based Cancer Registry were used for this study. The incidence of thyroid cancer was estimated for the area covered by the cancer registry. Survival of patients was ascertained by telephone call to the patients or the patient's family and/or linkage of registry data to mortality data from the Bureau of Vital Statistics. Patient's survival was based on sex, age and morphological type of tumour. RESULTS: Four hundred and twenty-nine cases of primary thyroid cancer were registered in the Tehran Metropolitan Area Cancer Registry. The incidence of thyroid cancer was 3.5 and 1.0 per 100,000 population per year for females and males, respectively. Seventy percent of tumours were papillary, 11% follicular, 6.2% medullary, and the rest were other subtypes. The papillary and follicular variants occurred in younger age: 43 +/- 16 and 46 +/- 13 years, respectively; the medullary and anaplastic variants occurred in older age: over 50 years. A 5-year survival rate was 82.2%, with median survival of 66 months and 95% confidence interval of 63 and 69 months. Men and women had a similar survival experience. CONCLUSIONS: While the incidence of thyroid cancer was slightly high, the descriptive epidemiology of thyroid cancer in Tehran did not manifest a unique feature. Tehran patients experienced a high rate of survival, and the survival time for males and females was similar. PMID- 16220014 TI - A clinic investigation into prehospital and emergency department delays in acute stroke care. AB - OBJECTIVE: The aim of our study was to investigate the factors which cause prehospital and emergency department (ED) delays in acute stroke care. SUBJECTS AND METHODS: We prospectively studied 229 acute stroke patients (median age: 71 +/- 19 years, 90 female and 139 male) who presented to the ED of the Gulhane Military Teaching Hospital, Istanbul, Turkey. Prehospital delay was defined as time from symptom onset to arrival at the ED. Emergency delay was defined as time from initial examination in the ED to arrival at the Neurology Intensive Care Unit. RESULTS: The median interval of prehospital and emergency delays were 92.66 and 53 min, respectively. The major cause of the prehospital delay was the time from symptom onset to first call for medical help (68.21 min, 73.93%, beta coefficients: 0.99; p < 0.001), and the major cause of the ED delay was waiting for the neurological consultation (21.28 min, 39.6%), beta coefficients: 0.03; p < 0.001). CONCLUSION: The results indicate that prehospital and ED delays are due to late decision to seek medical care and delayed neurological consultation. Hence, educational campaigns are needed to increase public awareness of stroke signs and the necessity of calling emergency services immediately when persons are suffering a possible stroke. Equally, ED physicians need to be trained in the recognition of symptoms and signs of acute stroke and the necessity for rapid neurological evaluation. PMID- 16220015 TI - In vitro activities of various antimicrobials against Brucella melitensis strains in the Aegean region in Turkey. AB - OBJECTIVE: To study in vitro activities of three quinolones (ciprofloxacin, levofloxacin, moxifloxacin), four macrolides (erythromycin, dirithromycin, azithromycin, clarithromycin) and doxycycline against 44 clinical isolates of Brucella melitensis. MATERIALS AND METHODS: Forty-four B. melitensis strains were isolated from blood cultures of adult patients with acute brucellosis who were hospitalized in the clinical ward of the Department of Clinical Microbiology and Infectious Diseases. The minimum inhibitory concentrations (MICs) of the tested antimicrobials were measured by the agar dilution method. MIC90 and MIC50 values were defined as the lowest concentration of the antibiotic at which 90 and 50% of the isolates were inhibited, respectively. RESULTS: Doxycycline (MIC50: 0.25 microg/ml, MIC90: 0.50 microg/ml) had the lowest MIC in vitro against the B. melitensis strains. Among the quinolones, ciprofloxacin and levofloxacin had similar activities (MIC50: 0.5 microg/ml, MIC90: 2 microg/ml), whereas MIC of moxifloxacin (MIC50: 1 microg/ml, MIC90: 8 microg/ml) was higher than both antibiotics in this group. Clarithromycin and azithromycin were the most active macrolides (MIC50: 8 microg/ml and MIC90: 32 microg/ml), followed by erythromycin (MIC50: 16 microg/ml, MIC90: 32 microg/ml) and dirithromycin (MIC50: 64 microg/ml and MIC90: 64 microg/ml). CONCLUSION: The results indicate that the conventional agent doxycycline is more active than quinolones and macrolides against the B. melitensis in vitro. PMID- 16220016 TI - Incidence of bloodstream infections in a speciality hospital in Kuwait: 8-year experience. AB - OBJECTIVES: To determine the frequency of isolation and antibiotic-susceptibility patterns of clinically significant bacterial pathogens isolated from blood. MATERIALS AND METHODS: The study was conducted over a period of 8 years (1995 2002) at Infectious Diseases Hospital (IDH), Kuwait. Demographic and clinical data were obtained from medical records. 18,535 blood cultures were analyzed. Disk diffusion method was used to perform antibiotic-susceptibility testing. Minimum inhibitory concentrations of 9 antimicrobials were determined using E test. Double disk (potentiation) test and E-test ESBL strips were used to detect the production of extended-spectrum beta-lactamases (ESBLs). RESULTS: Salmonella spp. and Brucella spp. were predominant blood isolates, and represented 60.6 and 30.0% of all clinically significant episodes of bloodstream infections, respectively. Among the Salmonella, Salmonella enterica serotypes typhi and paratyphi A were most frequently isolated. The percentage of multidrug resistance (MDR) among them varied from 22 to 51%. A high percentage (40%) of MDR S. enterica serotypes typhi and paratyphi A also showed reduced susceptibility to ceftriaxone and ciprofloxacin. CONCLUSION: During the study period, Salmonella spp. and Brucella spp. were predominant blood isolates. MDR S. enterica serotypes typhi and paratyphi A, with reduced susceptibility to ceftriaxone and ciprofloxacin, are among the most frequent causes of bloodstream infections in IDH, suggesting the need to monitor their susceptibility. PMID- 16220018 TI - Pulmonary arteriovenous malformation presenting with severe hypoxemia. AB - OBJECTIVE: To report a case of right pulmonary arteriovenous malformation (PAVM) affecting the right upper lobe, following the incidence of empyema thoracis in the contralateral lung. CLINICAL PRESENTATION AND INTERVENTION: A 19-year-old, previously healthy male presented with acute respiratory distress, left pleuritic chest pain, fever and hypoxemia. Clinical findings, laboratory and radiological examinations including pulmonary angiogram were consistent with the diagnosis of left pneumonia complicated with parapneumonic pleural effusion and right upper lobe PAVM. The patient was intubated and ventilated because of persistent hypoxemia. He was successfully treated by percutaneous transcatheter embolization. CONCLUSION: This case shows that percutaneous transcatheter embolization is a safe and effective first option for the treatment of PAVM. PMID- 16220017 TI - Modulation of the transforming growth factor beta1 by vitamin E in early nephropathy. AB - OBJECTIVE: The aim of this study was to investigate the pharmacological activity of an antioxidant, alpha-tocopherol (vitamin E, VE) in streptozotocin-induced diabetic rats and study its role in modulating the transforming growth factor beta1 (TGF-beta1). METHODS: Male Sprague-Dawley rats were treated with streptozotocin to induce diabetes. VE and/or insulin (INS) were administered daily during treatment periods of 3, 5, 7 and 10 days. Plasma glucose and fructosamine were measured in diabetic rats at the end of each treatment period. Samples of plasma, urine and renal cortex were analyzed for changes in protein and lysozyme excretion, reduced glutathione and malondialdehyde formation. TGF beta1 was determined by ELISA and expression of TGF-beta1 mRNA was investigated by RT-PCR and Northern blot analysis. RESULTS: Diabetes-induced glycemic stress was suppressed by INS, VE or a combination of INS and VE. Diabetes-induced increases of glucose, protein and lysozyme excretion were markedly depressed after 10-day treatment with INS, VE and the combination of INS and VE. Decreased glutathione content in the renal cortex of diabetic rats recovered towards control values, especially after 10-day treatment. Malondialdehyde content increased in diabetic rats and was reduced towards control value following 7- and 10-day treatments. Treatment of diabetic rats with INS, VE or the combination of INS and VE decreased elevated TGF-beta1 in plasma, decreased excretion of TGF beta1 in urine, and decreased renal cortex TGF-beta1 mRNA levels. CONCLUSIONS: Diabetes-induced overexpression of TGF-beta1 mRNA was suppressed by VE and INS after 5-, 7- and 10-day treatments. The results obtained with the antioxidant VE suggest that oxidative stress is involved in the development of diabetic nephropathy. Therefore, VE treatment may be effective in early stages of diabetic nephropathy to decrease or prevent pathological complications. PMID- 16220019 TI - Abdominal wall endometriosis: a diagnostic dilemma for surgeons. AB - OBJECTIVE: To report 3 cases of endometriosis of the abdominal wall, a disease which is unfamiliar to general surgeons because of the potential pitfalls in its diagnosis. CLINICAL PRESENTATION AND INTERVENTION: Three patients were referred to our general surgery clinic for abdominal masses. Incisional hernia and an abdominal mass were initially suspected in 2 patients, while a preoperative diagnosis of a rectus abdominis hematoma was made in the third because she had no history of previous surgery. Pain was a remarkable complaint in only one of the present cases. Abdominal wall endometriosis was diagnosed only upon histological examination postoperatively. In all cases, ultrasonography revealed hypoechogenic masses, and computed tomography showed that these masses had spiculations, and macroscopic views of the resected masses revealed well-demarcated margins without peritoneal involvement. All patients were treated with wide radical resections followed by polytetrafluoroethylene patch grafting. They were discharged from hospital on either the 2nd or the 3rd postoperative day uneventfully, and during follow-up there were no signs of pelvic endometriosis, as confirmed by ultrasonography, CA 125 measurement, gynecological consultation and examination. CONCLUSION: Since the diagnosis of scar endometrioma is rarely established prior to surgery, endometriosis should be included in the differential diagnosis of masses on the abdominal wall. PMID- 16220020 TI - Nonalcoholic Wernicke's encephalopathy with prominent astasia and optic neuropathy. AB - OBJECTIVE: To present a case with nonalcoholic Wernicke's encephalopathy (WE) developing astasia and optic neuropathy as major sequelae. CLINICAL PRESENTATION AND INTERVENTION: A 47-year-old woman developed WE following operation for pyloric stenosis. She received total parenteral nutrition before and after operation, but on the second postoperative day she developed visual hallucination and confusion, followed by nystagmus, ophthalmoplegia, apathy, dysarthria and coma. Although the patient has recovered with thiamine treatment, astasia and optic neuropathy persisted s major morbidities. CONCLUSION: The report shows that astasia and optic neuropathy may be prominent sequelae in some patients with WE. PMID- 16220021 TI - Complementary and alternative medicine: an academic view. PMID- 16220022 TI - Effects of tobacco smoking on the kinetics of the pupillary light reflex: a comparison between smokers and non-smokers. AB - BACKGROUND: The time course of the pupillary light reflex (PLR) is determined by the successive activation of parasympathetic and sympathetic innervations of the iris, latency and amplitude reflecting parasympathetic activity and recovery time showing mainly sympathetic activity. OBJECTIVE: To determine the effects of tobacco cigarette smoking on the PLR in smokers after an abstinence period of at least 12 h. METHODS: Ten smokers (mean 15.7 cigarettes/day) and 10 non-smokers participated in a randomised, non-intervention controlled, cross-over study that included a parallel control group. Smokers underwent two sessions with a time interval between 3 and 8 days; two recordings were taken at each session, separated by 20 min: session 1, without smoking, and session 2, smoking 3 cigarettes within a 30-min period. Non-smokers underwent one session; two recordings were taken separated by 20 min. At each recording, in both groups, PLR was elicited with four light flashes of increasing luminance. RESULTS: The relationship between PLR parameters and light intensity was linear in each subject. The slope of the regression line for relative amplitude increase versus intensity was significantly flatter in abstinent smokers than in non-smokers (p=0.033); the slope returned significantly after smoking (p=0.043). No other significant effects were obtained. CONCLUSIONS: Kinetic parameters of PLR provide a sensitive pharmacological test to detect cholinergic neurotransmission manipulation effects, as they seem to detect changes in moderate smokers after 12 h of abstinence, and their reversal on return to smoking. These results suggest an enhancement in the suppression of the parasympathetic oculomotor reflex arc rather than a facilitation of the sympathetic drive to the iris. PMID- 16220023 TI - Obsessive-compulsive disorder, factor-analyzed symptom dimensions and serotonin transporter polymorphism. AB - Recently, on the basis of the effects of serotonin (5-HT) reuptake inhibitors in obsessive-compulsive disorder (OCD) treatment, several candidate genes related to 5-HT regulation have been hypothesized to play an important role in the development of OCD. One of them is 5-HT transporter gene. Therefore, the aims of this study were to investigate the associations between 5-HT transporter polymorphism and OCD. One hundred and twenty-four OCD patients and 171 normal controls participated in this study. Genomic DNA was extracted from their blood. Comparison of the genotypes and allele frequencies of the SERTPR polymorphism between the OCD group and the control group was made. Using principal component analysis, we derived four factors from thirteen main contents of the Y-BOCS checklist and investigated the association between these four factors and the SERTPR polymorphism. In this case-control study, we could not find any associations between the SERTPR polymorphism and the development of OCD. In the OCD group, patients with the L genotype had higher scores for the religious/somatic factor than with the S genotype. In conclusion, the SERTPR polymorphism does not affect the development of OCD. But SERTPR polymorphisms affect certain factors of OC symptoms. Moreover, the factor analytic approach used in the present study has identified meaningful symptom dimensions to help guide future research. PMID- 16220024 TI - Interictal slow-wave focus in left medial temporal lobe during bilateral electroconvulsive therapy. AB - The interictal state between two electroconvulsive therapy (ECT) sessions is clinically characterised by possible cognitive adverse effects like mild amnestic syndrome. ECT-induced amnestic deficits can persist for several weeks after ECT. Electrophysiologically, slowing of brain electrical activity in the interictal state has often been reported. Especially, for bilateral ECT a correlation between enhanced left frontotemporal theta activity and retrograde amnesia has been demonstrated. This study focuses on the topographic distribution of cortical slow-wave oscillations during the interictal state of a bilateral ECT cycle. Twelve patients with major depression have been investigated with 32-channel resting EEG 24 h after the 6th ECT session. As controls, 8 major depressive patients were investigated prior to antidepressive treatment. The generating sources of slow-wave activity are estimated within the theta frequency band with low-resolution brain electromagnetic tomography. Source analysis revealed a distinct pattern of theta activity in the depth of the left temporal lobe (fusiform and parahippocampal gyri, Brodmann areas 37 and 36, respectively; p< 0.05) during the interictal state. This finding suggests a dysfunction of the left medial temporal lobe memory system during the interictal state of a bilateral ECT cycle. It will further be discussed whether it is possible to obtain information about activity of deep brain structures like the hippocampal formation from scalp-recorded signals. PMID- 16220025 TI - Antioxidant and nitric oxide-sparing actions of dihydropyridines and ACE inhibitors differ in human endothelial cells. AB - The effects of dihydropyridine Ca2+ channel blockers (DHP) and ACE inhibitors on superoxide formation and nitric oxide (NO) bioavailability were compared in human EA.Hy926 endothelial cells (EC). EC were stimulated 4 h with angiotensin II (Ang II, 10 nM) +/- study drugs. Specific superoxide formation was measured by lucigenin-enhanced chemiluminescence, reduction of cytochrome c and rhodamine-123 fluorescence. Free NO release was determined with an amperometric NO sensor. NADPH oxidase subunits expression was examined with Western Blot. In untreated EC the intracellular superoxide is -64.3 +/- 6.0% decreased compared to Ang II stimulated EC. Elevated extracellular superoxide formation was on a -43.0 +/- 1.7% lower level in untreated EC. The DHP Ca2+-channel agonist BayK8644 and ACE inhibitors captopril and ramiprilat led extracellular superoxide concentration to control level. Enalaprilat blocked extracellular superoxide, the DHP amlodipine and nisoldipine prevented intracellular increases only (n = 8-9, p < 0.05). Icatibant (HOE 140), a kinin-B2 receptor antagonist, attenuated antioxidant actions of all tested agents except of nisoldipine. Ang II-induced superoxide was elevated by the phorbolester PMA and blocked by the protein kinase C (PKC) inhibitor chelerythrine. Suppression of substance P-evoked NO release by Ang II (>70%, n = 6) was reversed by the PKC inhibitor chelerythrine, the DHP amlodipine and nisoldipine and the ACE inhibitor ramiprilat. Further, Ang II reduces Nox-4 expression by 34.5 +/- 4.9. Nox-2 expression was not regulated. DHP and ACE inhibitors exert different antioxidant effects in human EC stimulated with Ang II, but both improve NO bioavailability via bradykinin and modulation of redox regulating enzymes. PMID- 16220026 TI - [Postnatal follow-up of infants born to mothers with certain Toxoplasma gondii infection: evaluation of prenatal management]. AB - The clinical management of perinatal toxoplasmosis involves a gynaecologist during pregnancy and a neonatologist after delivery. Then, in the absence of a uniform approach, early evaluation of infected infants requires a thorough long term follow-up also in asymptomatic children, who have to be observed for at least one year due to unpredictable sequelae in later life. We retrospectively analyzed pregnancy management of 54 women with certain infection from Toxoplasma gondii (TG) and prospectively enrolled their infants to compare prenatal management with postnatal clinical outcome. All mothers with seroconversion for TG infection were from the Palermo area and were retrospectively analyzed, whereas their newborns referred to G. Di Cristina Children Clinical Hospital between 1999-2004 were prospectively enrolled in a 48-month follow-up. Timing of infection was dated for 24 women (45%) to the first trimester, 18 (33%) to the second and 12 (22%) the third. The maternal-fetal transmission rate was 17.2%. Prenatal diagnosis from amniotic fluid was performed in 25/54 pregnant subjects and showed positive results in 6. Despite diagnosis of TG infection, 9 women were untreated and only 2 with positive amniocentesis received combined therapy. 10/55 enrolled infants were infected and half of them were preterm and/or SGA at birth. None showed peculiar signs of TG at birth but 4 had abnormalities during the follow-up. 9/10 infected children were born to mothers who had undergone neither amniocentesis nor combined therapy. CONCLUSIONS: Our work confirms the difficulty of applying standardized therapeutic protocol for TG infection during pregnancy. The asymptomatic course of TG infection at birth confirms the importance of an instrumental long-term follow-up to identify typical TG lesion to prevent sequelae. PMID- 16220027 TI - Prevalence of antibodies to HHV-8 in the general population and in individuals at risk for sexually transmitted and blood-borne infections in Catania, Eastern Sicily. AB - BACKGROUND: Human herpes virus 8 (HHV-8) is the putative infectious agent of multifactorial diseases, such as Kaposi's sarcoma (KS), primary effusion lymphoma and multicentric Castleman's disease. However, its exact mode of action as well as its transmission is still under investigation. Besides, little is known about its seroprevalence in the population. HHV-8 epidemiology has been widely studied all over the world, demonstrating significant differences in distribution among various geographical areas and various population communities. Very few studies of HHV-8 seroprevalence have been conducted in Italy, particularly in Sicily which, along with other Mediterranean areas, is known to have high rates of KS incidence. Between January 2001 and April 2002, 424 patients were consecutively recruited from three treatment facilities. An Infectious Diseases Clinic provided 196 anti-HIV positive patients, both affected by AIDS and not. A further 122 anti HIV negative intravenous drug users were recruited from drug treatment clinics, while as a control group from the Blood Bank 126 blood donors were recruited. Base-line serum samples were assayed for antibodies to HHV-8 latency-associated nuclear antigen (anti-LANA) by IFA (Viramed Biotech AG, Planneg/Steinkirken- Germany). Anti-HHV-8 antibodies were found in 98 individuals (23.1%). HHV-8 reactivity was more common among anti-HIV positive patients (89/196, 45.4%, 95 C.I. 38.4-52.4) than in IDUs (6/102, 5.9%, 95 C.I. 1.2-16.2) and the control group (3/126, 2.4%, 95 C.I. 0.7-10.1). Overall, anti-HHV-8 antibodies were found in all three groups with large differences between groups. PMID- 16220028 TI - Hepatitis B virus DNA quantitation and detection of core promoter, precore and polymerase mutations in chronic hepatitis B: evaluation and clinical usefulness of three new commercial assays. AB - BACKGROUND: HBV-DNA quantitation, the HBe antigen status and the appearance of mutations in the core promoter, precore and polymerase regions are important elements in the management of chronic HBV infection. METHODS: We performed a technical evaluation of 3 new kits, affigene HBV VL, affigene HBV mutant VL and affigene HBV DE/3TC assays (Sangtec Molecular Diagnostics) in comparison with the Amplicor HBV Monitor assay (Manual Test, Roche), direct sequencing and direct sequencing/Inno-LIPA HBV DR (Innogenetics), respectively. We evaluated the clinical application of these tests in the management of patients with chronic (HBeAg positive) hepatitis B. Serial sera of 11 chronic HBeAg positive patients were studied before, during and after lamivudine/interferon treatment. RESULTS: HBV-DNA quantitation detected with affigene HBV VL showed a high correlation with the Amplicor HBV Monitor test (r=0.85). affigene HBV mutant VL (positions G1764A, G1896A) and affigene HBV DE/3TC (positions rtL180M, rtM204V/I) were able to detect a low presence of mutants in a mixed population (wild type and mutant) compared to direct sequencing and Inno-LIPA HBV DR, which identified only the dominant population. CONCLUSIONS: These three sensitive assays, performed with the same DNA extraction, give clinicians useful information for the management of chronic hepatitis B and for timing treatment. PMID- 16220029 TI - In vitro activity of fosfomycin in combination with vancomycin or teicoplanin against Staphylococcus aureus isolated from device-associated infections unresponsive to glycopeptide therapy. AB - Fosfomycin is a molecule that inhibits the early stage of peptidoglycan synthesis and shows a broad-spectrum bactericidal activity against Gram-positive and Gram negative bacteria. Using the Killing-curve method, we tested the in vitro bactericidal activity of fosfomycin alone or in combination with vancomycin or teicoplanin at a concentration of 8 microg/mL, that is easily achievable in serum at standard dosing regimens, against seven methicillin-resistant Staphylococcus aureus strains, isolated from patients with well documented device-associated infections unresponsive to or relapsing after glycopeptide therapy. MICs of vancomycin ranged from 1 to 4 microg/mL, MICs of teicoplanin from 2 to 8 microg/mL; MICs of fosfomycin were 8 microg/mL for two strains and >128 microg/mL for the remaining strains. The seven strains proved tolerant when tested for vancomycin and teicoplanin used alone at 2x MIC concentration. Fosfomycin was bactericidal (reduction of 2 log of the inoculum) only against the two susceptible strains. In all cases both vancomycin and teicoplanin in combination with fosfomycin developed bactericidal synergism already at a concentration of 1x MIC. If these results are confirmed by in vivo experiments, the combination of fosfomycin with glycopeptides might be useful for treating device-associated infections, and in preventing the phenomenon of increasing MICs for glycopeptides. PMID- 16220030 TI - [Prevalence and risk factors for bacteriuria in patients with cirrhosis]. AB - Bacterial infections occur frequently in patients with cirrhosis and may worsen the disease outcome. We investigated the prevalence of bacteriuria in 500 consecutive patients with cirrhosis, in different Child-Pugh stages (41.4% A; 40.8% B; 17% C) and analysed the associated risk factors. Most of the cirrhosis cases were virus related; alcohol abuse was recorded in 6.2% of the patients. Bacteriuria was detected in 139 (27.8%) cases: 32.4% were more than 100,000 cfu/ml; 7.9% between 100,000 and 1.000,000 cfu/ml and the remaining cases more than 1000,000 cfu/ml. Escherichia coli was the most frequent isolated agent (84.5%); Proteus spp. strains were detected only in bacteriuria with more than 100,000 cfu/ml. At univariate analysis, female gender, age and presence of diabetes were significantly associated to bacteriuria, while Child-Pugh stage and the presence of hepatocellular carcinoma were not. In a multivariate model, only female gender and diabetes were significantly associated to bacteriuria. These results indicate that advanced cirrhosis was not a risk for bacteriuria, that was associated rather to gender and diabetes, which are common risk factors for bacteriuria in non-cirrhotic patients. PMID- 16220031 TI - [Cutaneous myiasis from Cordylobia anthropophaga in a traveller returning from Senegal: a case study]. AB - Myiasis is the infestation of human or animal tissues by fly larvae. The disease is widespread especially in tropical countries. Here we report a case of myiasis due to Cordylobia anthropophaga that occurred in a traveller returning from Senegal. This case has some peculiar characteristics, regarding the site of the lesion and the clinical presentation. PMID- 16220032 TI - Linezolid in the treatment of severe central nervous system infections resistant to recommended antimicrobial compounds. AB - The progressive emergence of antimicrobial-resistant Gram-positive cocci especially in the setting of surgery and intensive care, recommends particular attention in making sound therapeutic choices to overcome both microbial resistances and haemato-encephalic barriers to effective local drug penetration. As in other Western countries, the occurrence of methicillin-resistant Staphylococcus aureus is particularly high also in Italy, especially when high risk patients and/or settings are involved. In treating post-neurosurgical central nervous system infections (cerebral abscess and meningitis), a key issue is represented by the low cerebrospinal fluid concentration of the two available glycopeptide antibiotics (vancomycin and teicoplanin), usually recommended as first-line therapy of resistant Gram-positive cocci. Recent findings have focused on the possible role of linezolid, an oxazolidinone antibiotic, as a suitable candidate for the treatment of severe brain infection (abscesses) and post neurosurgical infection, where treatment options and efficacy are significantly limited by the low glycopeptide transfer and the spread of glycopeptide-resistant bacterial strains. Three representative case reports (two brain abscesses and one post-surgical meningitis) are presented and discussed in light of the current literature: in all these cases, salvage linezolid treatment proved resolutory. PMID- 16220033 TI - [A case of ophthalmomyiasis: description and diagnosis]. AB - The authors describe a human case of ophthalmomyiasis caused by the "sheep fly" Oestrus ovis larva. They emphasize the importance of collaboration between practitioner or specialist and parasitologist for a sound, definitive aetiological diagnosis of such uncommon pathologies. PMID- 16220034 TI - [The fight against tuberculosis and developments in public health from 1890 to 1930 in Italy]. AB - The author presents the developments in society and public health that paved the way, thanks to the birth of social and scientific associations and later to state intervention, for the establishment of hospitals for tuberculosis in Italy at the beginning of the 20th century. From 1890 onwards many fruitful meetings, conferences and debates ultimately led to the passing of a specific law on public health care; in 1927 this law covered about half the population. The First World War was undoubtedly the prime cause that stopped the downward trend in mortality and morbidity from the tuberculosis epidemic in the early 20th century. Early on, Bologna, together with other cities in Tuscany and Lombardy, developed plans for therapy (i.e. hospitals for tuberculosis) and prevention (i.e. holiday camps). However, in Italy in the 1930s tuberculosis continued to be a major problem despite such improvements in public health. In Europe, Germany was the nation that pioneered the fight against tuberculosis by social, scientific and legislative means. Since 1883 there had been a specific law for insurance against illnesses and a network of sanatoria had been established. PMID- 16220035 TI - Infection and the chronic wound: a focus on silver. PMID- 16220038 TI - Gastric secretion. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the pertinent literature published in the past year regarding the regulation of gastric exocrine and endocrine secretion. RECENT FINDINGS: Gastric acid aids protein digestion; facilitates the absorption of iron, calcium, and vitamin B12; thwarts enteric infection; and prevents bacterial overgrowth. When levels of acid and proteolytic enzymes overwhelm the mucosal defense mechanisms, ulcers occur. To avoid damage under these harsh conditions, gastric acid must be finely regulated by overlapping neural (e.g. orexin, pituitary adenylate cyclase-activating polypeptide, nitric oxide, and galanin), hormonal (e.g. gastrin, cholecystokinin, and ghrelin), paracrine (e.g. histamine and somatostatin), and autocrine (e.g. transforming growth factor-alpha) pathways. The precise mechanisms whereby Helicobacter pylori induces perturbations in acid secretion are not known, but they seem to involve changes in somatostatin and perhaps ghrelin secretion. Acid secretion by parietal cells involves intracellular elevation of calcium and/or cyclic AMP, followed by a cascade that triggers translocation of the proton pump, HK-adenosine triphosphatase, from cytoplasmic tubulovesicles to the secretory canaliculi. SUMMARY: An improved understanding of the pathways and mechanisms regulating gastric acid secretion may lead to the development of new strategies to prevent and treat acid peptic disorders as well as circumvent the adverse effects of currently prescribed antisecretory medications. PMID- 16220039 TI - Interventional endoscopy. AB - PURPOSE OF REVIEW: This review is an update of key issues in gastric interventional endoscopy. It focuses on the areas of patient preparation, endoscopic mucosal resection, gastroduodenal stenting, and endoscopic placement of enteric feeding tubes. RECENT FINDINGS: Clopidogel (Plavix), a newer antiplatelet agent, can increase the risk of bleeding. Therefore, in selected cases, it should be held for 7-10 days prior to interventional procedures. In experienced hands, endoscopic mucosal resection (success rate, 76-100%; complication rate, 4-28%) and gastroduodenal stenting (success rate, 81-92%; complication rate, 1-17%) seem to be safe and effective techniques. SUMMARY: The field of interventional endoscopy continues to advance and to conquer new frontiers. These advances create new problems that need to be addressed and studied by researchers, however. It is only through these types of reviews that our state of knowledge can be updated to help provide the latest information for clinicians in the field and to challenge researchers with future problems that need to be studied. PMID- 16220040 TI - Helicobacter pylori virulence factors: facts and fantasies. AB - PURPOSE OF REVIEW: Virulence factors are related to the ability of a microbe to induce disease. True virulence factors must therefore have a disease association, an in-vivo correlate with disease such as increased mucosal inflammation, or both. RECENT FINDINGS: The cytotoxin-associated gene pathogenicity island; the outer membrane inflammatory protein; the duodenal ulcer-promoting gene, and possibly the blood group antigen-binding adhesion, are the only factors that to date qualify as virulence factors. Numerous recent studies have investigated the interaction of vacuolating cytotoxin A or cytotoxin-associated gene A with cells and cell lines in vitro. It remains unclear, however, whether any of the findings, for example, in-vitro experiments showing that vacuolating cytotoxin A affect the regulation of T or B lymphocytes, have an in-vivo counterpart, or play any role in disease pathogenesis. SUMMARY: The criteria for a virulence factor include evidence of an association with a disease or a disease surrogate such as the severity of mucosal inflammation, epidemiologic consistency, and biologic plausibility. Confirmation of the proposed mechanism requires elimination of the effect by gene deletion and restoration by complementation. Cytotoxin-associated gene A has been the subject of elegant biochemistry despite lack of evidence that it is involved in pathogenesis. The current focus of research on Helicobacter pylori relates to exploring the biology of Helicobacter pylori, often using systems that only vaguely relate to the in-vivo conditions or to disease pathogenesis. PMID- 16220041 TI - Cyclooxygenase-2 inhibitors. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize new information regarding the use of selective inhibitors of the cyclooxygenase-2 enzyme, emphasizing recent developments regarding cardiovascular risk. RECENT FINDINGS: Selective cyclooxygenase-2 inhibitors are as effective as nonselective nonsteroidal antiinflammatory drugs in relieving pain and inflammation but are associated with significantly fewer gastric, duodenal, and intestinal ulcers and ulcer complications. Cyclooxygenase-2 inhibitors may also reduce colorectal polyp development or recurrence as well as reduce the risk of colorectal and esophageal cancer. Some, but not all, studies have suggested increased myocardial infarction with certain cyclooxygenase-2 inhibitors, in particular rofecoxib. It is unclear whether this is a class effect because there are inconclusive data to incriminate celecoxib despite a relatively large number of clinical trials enrolling a large number of patients. Rofecoxib was voluntarily withdrawn by the manufacturer. The European Medicines Agency concluded that cyclooxygenase-2 inhibitors are contraindicated in patients with established cardiovascular disease, should be used with caution in patients with risk factors, and were justified in patients at risk for serious gastrointestinal adverse events. SUMMARY: Rofecoxib, but perhaps not all cyclooxygenase-2 inhibitors, may be associated with increased risk for myocardial infarction. It is unclear whether nonselective nonsteroidal antiinflammatory drugs increase or decrease the rate of myocardial infarction. The final truth awaits further studies. PMID- 16220042 TI - Gastric motor and sensory function. AB - PURPOSE OF REVIEW: Abnormalities of gastric sensory and motor function are considered key players in the pathogenesis of upper gastrointestinal symptoms in functional dyspepsia and in gastroparesis. This review summarizes recent progress in our understanding of normal and pathologic gastric sensory and motor function. RECENT FINDINGS: Several novel tests have been developed to study gastric sensitivity and meal-induced accommodation, including scintigraphic analysis of meal distribution within the stomach, single photon emission computed tomography of gastric volumes, and nutrient or water challenge tests. Several studies have reported on the occurrence of delayed gastric emptying in functional dyspepsia and in gastroparesis, but the correlation with symptoms was generally poor. The pathways and neurotransmitter involved in gastric hypersensitivity are progressively being unraveled. Several studies have confirmed and focused on increased symptom occurrence after nutrient challenge in functional dyspepsia. The role of the proximal stomach in the control of food intake and the stomach as a target in the treatment of obesity are areas of intense research. Studies have reported on pharmacologic approaches as well as electric stimulation in the treatment of gastric sensorimotor dysfunction. SUMMARY: Progress in our understanding of normal and abnormal gastric sensory and motor function may lead to new or improved treatment modalities. Areas of major advances are the study of meal-induced symptoms in functional dyspepsia, unraveling of the role of the stomach in the control of food intake, and the use of gastric electric stimulation in gastroparesis and in obesity. PMID- 16220043 TI - Chemotherapy and radiotherapy in the management of gastric cancer. AB - PURPOSE OF REVIEW: The treatment of locally advanced gastric cancer remains a major challenge. After decades of investigation that have yielded little improvement in survival rates, several important studies have recently emerged that provide renewed optimism for future research endeavors. This article reviews the important advances over the past several years in the use of chemotherapy and radiotherapy for gastric cancer. RECENT FINDINGS: As a result of the Intergroup trial, adjuvant chemoradiotherapy after operation has emerged as a new standard of care for patients who have undergone resection for carcinoma of the stomach. Strategies currently being tested to build on the Intergroup results include the use of new chemotherapy combinations with radiotherapy and the development of modern conformal techniques to deliver radiation therapy. The preliminary results of a large neoadjuvant chemotherapy study have demonstrated the efficacy of this approach with tumor downstaging and increase in the curative R0 resection rate. For patients with metastatic gastric cancer, newer generation cytotoxic agents such as oxaliplatin, irinotecan, and taxanes show promising activity. In the near future, these agents will likely be evaluated for their role as adjuvant and neoadjuvant therapy. SUMMARY: Major advances in the treatment of gastric cancer have occurred during the past several years and have improved the care of patients with this form of tumor. PMID- 16220044 TI - Surgery for obesity. AB - PURPOSE OF REVIEW: Bariatric surgery today is the only effective therapy for morbid obesity. Commonly performed procedures include adjustable gastric banding and vertical banded gastroplasty, variations of the Roux-en-Y gastric bypass, biliopancreatic diversion or duodenal switch, and mixed procedures. This review discusses key issues in the surgical management of morbid obesity. RECENT FINDINGS: The two most common bariatric procedures performed worldwide are laparoscopic adjustable gastric banding and laparoscopic Roux-en-Y gastric bypass. Controversy exists regarding the best surgical procedure. Weight loss decreases according to the procedures performed in following decreasing order: biliopancreatic diversion, Roux-en-Y gastric bypass, vertical banded gastroplasty, adjustable gastric banding. Concerning the complications and quality of life, there is no single operation for morbid obesity without drawbacks. Cost-effectiveness analyses have demonstrated that bariatric surgery is cost effective at less than 50,000 US dollars/quality-adjusted life years. SUMMARY: According to current opinion, gastric restrictive procedures (adjustable gastric banding, vertical banded gastroplasty) are generally considered safe and quick to perform, but the long-term outcome and quality of life have been questioned. By contrast, the long-term efficacy of adjustable gastric banding can be improved by the development of new band devices. More complex bariatric procedures, such as the Roux-en-Y gastric bypass or biliopancreatic diversion, have a greater potential for serious perioperative complications but are associated with good long-term outcome in terms of weight loss combined with less dietary restriction. PMID- 16220045 TI - Immunology. PMID- 16220046 TI - Lamina propria dendritic cells in the physiology and pathology of the gastrointestinal tract. AB - PURPOSE OF REVIEW: The mucosal immune system constantly surveys the intestinal microbiota. The outcome of this interaction is determined by the functional properties of dendritic cells, which play a key role in immune response by facilitating antigen sampling and pathogen recognition, as well as innate host defenses. RECENT FINDINGS: Recent advances have provided insight into the diverse mechanisms involved in the acquisition of intestinal antigens and led to a new appreciation of organ-specific functional subspecification of dendritic cell subsets. Dendritic cells are not a rare cell type in the intestine but populate the entire lamina propria of the gastrointestinal tract as an extensive network. Distinct dendritic cell subsets may be associated with specific immune functions in the lamina propria and Peyer's patches. Newly discovered routes of antigen acquisition, such as the formation of transepithelial dendrites, allow dendritic cells direct access to the intestinal lumen to obtain information about commensal and pathogenic microorganisms. Dendritic cells emerge as key regulators of the intestinal immune system with their ability to direct intestine-specific migration and control of T cells. SUMMARY: Dendritic cells play a major role in the complex interactions between the gut microbiota and the innate and adaptive immune system, leading to tolerance and immunity. Recent contributions have yielded important information that may aid in the development of novel therapeutic approaches to the treatment of inflammatory bowl disease and intestinal infections and to new immunization strategies. PMID- 16220047 TI - Oral tolerance and inflammatory bowel disease. AB - PURPOSE OF REVIEW: Oral tolerance refers to the ability of the mucosal immune system to actively inhibit systemic immune responses to fed antigens. Recently, clinical trials have used oral tolerance as a therapy for certain chronic inflammatory and autoimmune diseases such as multiple sclerosis and type I diabetes. Inflammatory bowel disease is now widely thought to be caused by the breakdown of oral tolerance through a combination of genetic and environmental factors. Therefore, it seems incongruous that clinicians would try to use oral tolerance therapy to alleviate the symptoms of inflammatory bowel disease. Yet, armed with the results of select animal models, trials have begun for oral tolerance therapy for Crohn's disease. This review will outline the recent advances in understanding oral tolerance, explore the relation between oral tolerance and inflammatory bowel disease, and comment on the likelihood of successful oral tolerance therapy for inflammatory bowel disease. RECENT FINDINGS: The results of an oral tolerance trial in Crohn's disease patients in Israel have shown some promising results, whereas the results of studies of experimentally induced oral tolerance in patients with inflammatory bowel disease from the authors' laboratory have shown that feeding a neoantigen in an attempt to induce oral tolerance is not successful in patients with inflammatory bowel disease. SUMMARY: The fundamental difference in the mechanisms of oral tolerance in mice and humans requires a more focused effort to understand the human mucosal immune system before oral tolerance therapy for autoimmune and chronic inflammatory disorders reaches its full potential. PMID- 16220048 TI - Irritable bowel syndrome and probiotics: from rationale to clinical use. AB - PURPOSE OF REVIEW: Few therapies are of proven efficacy in irritable bowel syndrome. Thus, there is great interest in the development of a natural therapy that can be both safe and effective. An understanding that probiotics are heterogeneous, with multiple targets and mechanisms of action, is fundamental to the development of clinical trials. RECENT FINDINGS: A bidirectional model for the pathogenesis of irritable bowel syndrome is proposed in which gut-driven and brain-driven mechanisms contribute to the genesis of gut dysfunction and symptoms. In-vitro and animal studies have generated most of the mechanistic rationale for the use of probiotics in functional bowel disorders. A MEDLINE search of publications from 1989 to date revealed only eight placebo-controlled clinical trials on the subject of probiotics and irritable bowel syndrome. All these studies suffer from methodologic problems. By contrast, numerous reviews have been published in the past 2 years on this subject. SUMMARY: Animal research will continue to identify novel targets and elucidate the mechanisms of action of probiotics, thus providing a rational basis for their use in irritable bowel syndrome. The notion of treating irritable bowel syndrome with probiotics is particularly attractive to patients and generates great interest, although clinical evidence is not yet sufficient to enable clear guidelines to be designed. Large, well-designed, controlled clinical trials using specific probiotics are warranted. PMID- 16220049 TI - Role of inflammation in nonalcoholic steatohepatitis. AB - PURPOSE OF REVIEW: Products of hepatic macrophages and lymphocytes are acknowledged regulators of liver injury and repair. Recent studies have identified inflammatory modulators from sources within and outside the liver that are critical to the pathogenesis and progression of chronic liver diseases, including nonalcoholic fatty liver disease. This review will focus on these developments to clarify how inflammatory mediators from adipose tissue and the liver interact to mediate the pathogenesis of nonalcoholic fatty liver disease. RECENT FINDINGS: Hepatic steatosis and steatohepatitis are extremely prevalent in obese individuals with the metabolic syndrome. The metabolic syndrome results from abnormal production of various adipose-derived and liver-derived factors that modulate energy substrate flux to coordinate tissue anabolism and catabolism. Individuals with the metabolic syndrome produce a relative excess of proinflammatory factors. Some factors inhibit hepatic fat disposal and promote lipid accumulation within hepatocytes. The latter induces sustained hepatic generation of proinflammatory cytokines, particularly when the hepatic innate immune system becomes Th-1 polarized. Although chronic inflammation induces production of various profibrogenic factors, progression to latter stages of nonalcoholic fatty liver disease is relatively unusual in individuals with the metabolic syndrome. This may reflect requirements for additional factors that become abundant only in individuals who have additional defects in hepatic innate immunity. SUMMARY: Obesity and the metabolic syndrome represent chronic inflammatory states and are associated with nonalcoholic fatty liver disease. Liver injury that ensues is dictated by metabolic and immunomodulatory factors that are produced by adipose tissue and within the liver. PMID- 16220050 TI - Food allergy and irritable bowel syndrome. AB - PURPOSE OF REVIEW: Irritable bowel syndrome is a common and likely a multifactorial gastrointestinal disorder in which a disturbed brain-gut axis has been thought to have a mandatory role. Recent clinical and experimental studies imply that dietary factors may be more important in the pathogenesis of irritable bowel syndrome than was earlier anticipated. The purpose of this review is to present those studies and discuss their findings in relation to the crosstalk between the gastrointestinal immune and nervous systems. RECENT FINDINGS: Food elimination based on serum immunoglobulin G antibodies in irritable bowel syndrome has been found to result in a significant decrease in symptoms, compared with diets in which dietary restrictions are not guided by those antibodies. Both numbers of mast cells and their mediators have been shown to be increased in intestinal mucosa in patients with irritable bowel syndrome, especially in the close proximity of intestinal nerves. Animal studies have demonstrated that this increase in intestinal mast cell density could be a consequence of local hypersensitivity to food antigens. That kind of local gastrointestinal hypersensitivity seems to be beyond the reach of current diagnostic methods available in clinical practice. SUMMARY: Dietary factors may significantly contribute to the pathophysiology of irritable bowel syndrome. Elimination diets based on the detection of local food hypersensitivity may offer a treatment option for irritable bowel syndrome patients in the future. PMID- 16220051 TI - Synbiotics and the mucosal barrier in critically ill patients. AB - PURPOSE OF REVIEW: Outcome in severe and critical illnesses is strongly related to premorbid conditions: the strength of the mucosal barriers, the innate immune system, and the built-in resistance to disease. Early risk factors and determinants of poor outcome are factors such advanced age; impaired premorbid health status, especially diabetes and high body mass index (obesity); and immunosuppressive treatments. Combined supplementation of bioactive fibers and lactic acid bacteria (synbiotics) directly and indirectly influences several of these factors. RECENT FINDINGS: Determinants for poor outcome are degree of oxidative stress, neutrophil activation, and infiltration of tissues, especially in the lungs. Attempts at early reduction of the exaggerated inflammatory storm and limitation of further impairment of the immune function are always given the highest priority. The supply of live lactic acid bacteria and plant fibers can dramatically reduce the hyperinflammation and also the infiltration by neutrophils of organs such as the lungs. New and efficient autopositioning and regurgitation-resistant feeding tubes provide instruments for the early supply of enteral nutrition with immune-boosting antioxidants and synbiotics. SUMMARY: A meticulous choice of probiotic lactic acid bacteria is recommended because only a small minority of the lactic acid bacteria survive the harsh environment of the upper gastrointestinal tract, ferment strong semiresistant fibers such as inulin, and have the ability to control inflammation and eliminate unwanted pathogens, such as antibiotic-resistant microorganisms and Clostridium difficile. PMID- 16220052 TI - Bibliography. Current world literature. Stomach and duodenum. PMID- 16220053 TI - Why trust matters: the nurse manager-staff nurse relationship. PMID- 16220054 TI - Rethinking nurse recruitment: a return-on-investment approach. PMID- 16220055 TI - Ten recommendations for reducing unnecessary emergency department visits. PMID- 16220056 TI - The RWJ Executive Nurse Fellows Program, Part 1: Leading change. AB - This article is the first of a 3-part series describing the RWJ Executive Nurse Fellows Program, an advanced leadership program for nurses in senior executive roles who aspire to help lead and shape the US healthcare system of the future. This article describes the RWJ Executive Nurse Fellows program, its core leadership competencies, and the primary program components. Part 2 (December 2005) will focus on the nontraditional mentor relationships developed by fellows as part of the fellowship experience and lessons learned regarding best practices. Part 3 (February 2006) will explain how fellows are required to create a business plan for their leadership project because it is so important for nurse leaders to offer a strong business case for proceeding with a new initiative, service, or program. PMID- 16220057 TI - Empowering nurses for work engagement and health in hospital settings. AB - Employee empowerment has become an increasingly important factor in determining employee health and wellbeing in restructured healthcare settings. The authors tested a theoretical model which specified the relationships among structural empowerment, 6 areas of worklife that promote employee engagement, and staff nurses' physical and mental health. A predictive, non-experimental design was used to test the model in a random sample of staff nurses. The authors discuss their findings and the implication for nurse administrators. PMID- 16220058 TI - Excellence through evidence: securing collegial/collaborative nurse-physician relationships, part 1. AB - Multiple nurse-physician (RNMD) relationships coexist on hospital units; collegial and collaborative (C/C) relationships positively impact, more than others, patient outcomes. The goal of this multisite evidence-based management practice initiative was to identify structures that enable C/C RNMD relationships. In part 1, the authors discuss the methodology and selection of the sample of 141 physicians, managers, and staff nurses from 44 clinical units in 5 hospitals that had previously demonstrated extensive C/C RNMD relationships. These 141 experts were interviewed to identify structures enabling C/C RNMD relationships. Part 1 presents the structures that enable C/C RNMD relationships as described and tested in the literature, as well as a description of the characteristics of the clinical units, experts, and C/C RNMD relationships found in this study. In part 2, the structures identified by the experts as needed for securing C/C RNMD relationships will be presented, along with suggestions for attainment. PMID- 16220059 TI - Nurse staffing and patient outcomes in Veterans Affairs hospitals. AB - OBJECTIVE: To assess characteristics and perceptions of nurses working in the Veterans Health Administration (VHA), comparing types of nursing personnel, to benchmark to prior studies across healthcare systems. BACKGROUND: Prior studies have shown relationships between positive registered nurse (RN) perceptions of the practice environment and patient outcomes. To date, no study has reported the comparison of RN perceptions of the practice environment in hospital nursing with those of non-RN nursing personnel. This study is the first to offer a more comprehensive look at perceptions of practice environment from the full range of the nursing work force and may shed light on issues such as the relationship of skill mix to nurse and patient outcomes. METHODS: Cross-sectional observational study with a mailed survey administered to all nursing personnel in 125 VA Medical Centers between February and June 2003. RESULTS: Compared with other types of nursing personnel in the VHA, RNs are generally less positive about their practice environments. However, compared with RNs in other countries and particularly with other RNs in the United States (Pennsylvania), VHA RNs are generally more positive about their practice environment and express more job satisfaction. CONCLUSIONS: The nursing work force of the VHA has some unique characteristics. The practice environment for nurses in the VHA is relatively positive, and may indicate that the VHA, as a system, provides an environment that is more like magnet hospitals. This is significant for a public sector hospital system. PMID- 16220060 TI - Organizational commitment to professional practice models. AB - Magnet hospitals must show evidence of professional practice models. A professional practice organizational environment is necessary, but not sufficient, to create professional practice models. The authors analyze the relationship between a professional practice environment and a professional practice and explore organizational commitments required to ensure professional practice at the unit level. PMID- 16220061 TI - Dose-dependent autonomic dysfunction in chronic L-NAME-hypertensive diabetic rats. AB - This study investigated the effects of varying doses of L-NAME on arterial pressure (AP), baroreflex control, and heart rate (HR)/AP variability in the STZ diabetic rat. Fifty-two male Wistar rats were injected with 50 mg/kg IV STZ (diabetes, D, n = 24) or citrate (controls, C, n = 28) 30 days before recordings. After 16 days, they received 14 days of oral L-NAME, 10 (H10) or 30 (H30) mg/kg, or water. Catheters were implanted into the femoral artery and vein (PE-10) for measurements in conscious rats; recorded data were analyzed on a beat-to-beat basis. Mean AP was higher in CH30 versus C and in DH10 and DH30 versus D rats. Reflex tachycardia was blunted in CH30 and DH30 rats (b = -1.81, -1.41, -0.48 in C, CH10, and CH30, respectively, P < 0.05 and b = -1.45, -1.19, -0.28 in D, DH10, and DH30, respectively, P < 0.05). Although HR and AP variability were reduced in CH30 and DH30 rats versus C and D rats, the DH30 rat had more accentuated dysfunction. All doses of L-NAME produced similar AP responses in experimental versus control groups, independent of the disease state (diabetes). Thus, autonomic dysfunction is more related to the L-NAME dose used and to the association of diabetes and hypertension than to AP values. PMID- 16220062 TI - Bioflavonoids effectively inhibit smooth muscle cell-mediated contraction of collagen matrix induced by angiotensin II. AB - Plant-derived bioflavonoids have been recognized to support arterial wall structural integrity and interfere with a variety of proatherosclerotic stimuli. In this study we tested the effects of bioflavonoids on the contractile activity of cultured human aortic smooth muscle cells (SMC) embedded in a 3-dimensional type I collagen matrix. Collagen I solution mixed with human aortic SMC in 24 well plates were allowed to form gels. Tested compounds were added to the wells, and the gels were set afloat by gentle tapping. Digital photographs of the gels were taken after 24 hours of incubation at 37 degrees C. The area of contracted gel was measured and expressed as a percentage of the control gel area from 3 or more replicates. Expression of matrix metalloproteinase (MMP-2) in conditioned media was assessed by gel zymography. Different classes of bioflavanoids showed variable efficiency in inhibiting angiotensin II (ATII)-dependent collagen gel contraction by SMCs. An increase in the number of gallate groups per catechin molecule was associated with increased inhibition of angiotensin II-dependent collagen gel contraction by SMC. Antioxidants (N-acetyl cysteine and ascorbic acid) did not inhibit collagen gel contraction. Bioflavonoid inhibition of collagen gel contraction by SMC correlated with inhibition of matrix metalloproteinase-2 expression. Bioflavonoids participate in the regulation of SMC-mediated contraction and have a strong potential in counteracting pathophysiological effects of ATII. Bioflavonoid activity depends on structural characteristics and can be related to extracellular matrix integrity. PMID- 16220063 TI - Pharmacokinetics and biodistribution of a pGT2-VEGF plasmid DNA after administration in rats. AB - Intramyocardial administration of gene therapy vectors expressing angiogenic factors have been attempted as an alternative to conventional surgical methods for the management of myocardial ischemia. In this study, we have developed the pGT2-VEGF, a plasmid DNA vector expressing human VEGF165, for the management of ischemic cardiovascular disease and investigated in vivo pharmacokinetics and tissue distribution of pGT2-VEGF after intramyocardial and intravenous administration in rats. A high concentration of pGT2-VEGF was observed in the heart after intramyocardial injection of 300 microg, which is in line with the assumption that direct intramyocardial delivery enables extended localization at the administration site. Leakage of the pGT2-VEGF to the blood circulation was observed after intramyocardial injection, with an area under the curve (AUC) of 3.8 microg min/mL, as compared with 37.3 microg min/mL after intravenous injection of the same dose. The pGT2-VEGF concentration in blood peaked at 5 minutes after intramyocardial administration and declined rapidly to undetectable levels by 2 hours post-administration. In tissue distribution studies, pGT2-VEGF peaked at 5 minutes post-administration in various organs but was undetectable at 2 hours in all organs except heart, lung, and liver. Taken together, the results suggest that intramyocardial-delivered pGT2-VEGF was degraded rapidly in vivo and mainly persisted in target tissues, the heart. In addition, intramyocardial administered pGT2-VEGF was expressed for longer periods than the persistence of the pGT2-VEGF plasmid DNA in a target tissue. Therefore, a direct myocardial injection of pGT2-VEGF might be useful for local therapeutic angiogenesis. PMID- 16220064 TI - A review of the structural and functional features of olmesartan medoxomil, an angiotensin receptor blocker. AB - The angiotensin II (A-II) type 1 (AT1) receptor-mediated effects of A-II play a key role in the pathophysiology of hypertension. Effective inhibition of A-II is provided by the latest class of antihypertensive medications, the AT1 receptor blockers (ARBs). These orally available agents were developed around a common imidazole-based structural core. The most recent member of this drug class to be approved by the Food and Drug Administration, olmesartan medoxomil, contains unique features that may explain its clinical efficacy. Key structural elements of olmesartan medoxomil include a hydroxyalkyl substituent at the imidazole 4 position and a hydrolyzable ester group at the imidazole 5-position. Inter- and intramolecular hydrogen bonding involving these groups may contribute to the potentiation of antagonist activity. After oral administration, olmesartan medoxomil is deesterified in the intestinal tract to produce the active metabolite olmesartan, which undergoes no additional metabolic change. The marked antihypertensive efficacy of olmesartan medoxomil may result from a unique pharmacological interaction of the drug with the AT1 receptor, resulting in a potent, long-lasting, dose-dependent blockade of A-II. This review article characterizes the structural features of olmesartan that may be responsible for its clinical efficacy. Inferential pharmacological studies compare and contrast the effects of olmesartan to those of other ARBs in comparable preclinical animal models. PMID- 16220065 TI - Use of a treatment algorithm to achieve NCEP ATP III goals with atorvastatin. AB - This multicenter, 8-week, single-step titration, open-label study sought to assess the percentage of subjects who achieved their National Cholesterol Education Program Adult Treatment Panel (NCEP ATP) III low-density lipoprotein (LDL)-cholesterol target when assigned a starting dose of atorvastatin (10 mg, 20 mg, 40 mg, or 80 mg) selected using an algorithm based on their 10-year CHD risk and the magnitude of LDL-cholesterol lowering necessary to reach goal. Following an 8-week washout period, 1298 subjects, categorized as low, medium, or high risk, were assigned to 4 weeks of treatment with a starting dose of atorvastatin selected by the algorithm. At week 4, subjects who did not achieve goal (15.8%) were titrated to the next-higher dose. The primary endpoint was the percentage of subjects in each risk group who achieved LDL-cholesterol goal at week 8. At 8 weeks, 84.8% of subjects (low risk 92.9%; moderate risk 84.0%; high risk 81.1%) achieved LDL-cholesterol target. The majority of patients (84.2%) achieved their lipid target through the use of the algorithm-based starting dose (10-80 mg) without the need for titration. No patient had elevations in creatine phosphokinase >10 times the upper limit of normal. Elevations in alanine aminotransaminase or aspartate aminotransaminase were observed in <1% of study subjects and were unrelated to dose. Selecting the starting dose of atorvastatin using a treatment algorithm achieves NCEP ATP III LDL-cholesterol goals in the majority of patients and minimizes the need for dose titration. PMID- 16220066 TI - Aortic smooth muscle relaxants KMUP-3 and KMUP-4, two nitrophenylpiperazine derivatives of xanthine, display cGMP-enhancing activity: roles of endothelium, phosphodiesterase, and K+ channel. AB - The cellular mechanisms of vasorelaxant effects of newly synthesized KMUP-3 and KMUP-4 were investigated in rat aortic smooth muscle (RASM). KMUP-3 (7-[2-[4-(4 nitrobenzene)piperazinyl]ethyl]-1,3-dimethylxanthine) and KMUP-4 (7-[2-[4-(2 nitrobenzene)piperazinyl]ethyl]-1,3-dimethylxanthine) elicited concentration dependent relaxation of endothelium-intact and denuded RASM precontracted with phenylephrine. Relaxant responses were also produced by the PDE inhibitors theophylline, milrinone, rolipram, and zaprinast (1 nM-100 microM). The relaxant responses of KMUP-3 and KMUP-4 were reduced by endothelium removal and by the presence of the NOS inhibitor L-NAME (100 microM), the sGC inhibitor ODQ (1 microM), the adenylyl cyclase (AC) inhibitor SQ 22536 (100 microM), and the prostaglandin inhibitor indomethacin (10 microM). Additionally, the vasorelaxations of both agents were also attenuated by pretreatment with the nonselective K+ channel blocker TEA (10 mM), the KATP channel blocker glibenclamide (1 microM), the voltage-dependent K+ (KV) channel blocker 4-AP (100 microM), and Ca(2+)-dependent K+ (KCa) channel blockers apamin (1 microM) and charybdotoxin (ChTX, 0.1 microM). In addition, elevated extracellular K+ (80 mM) interferes with KMUP-3- and KMUP-4-induced vasorelaxations. Preincubation with both agents (1 microM) significantly enhanced the dilator responses of isoproterenol and SNP. KMUP-3 and KMUP-4 inhibited PDE activities and increased cAMP and cGMP levels in primary culture of RASM that were inhibited by SQ 22536 and ODQ, respectively. In cultured HUVECs, KMUP-3 and KMUP-4 (0.1 microM), more potent than YC-1, significantly increased the expression of eNOS protein. In summary, KMUP-3 and KMUP-4 induce aortic relaxations through both endothelium dependent and -independent mechanisms. Mechanisms of vasorelaxation induced by both compounds involve multiple processes, such as accumulation of cyclic nucleotides partly as a result of PDE inhibition, K-channel activation, and indomethacin-sensitive endothelium function. PMID- 16220067 TI - Comparison of two doses and dosing regimens of tolvaptan in congestive heart failure. AB - Fluid retention and extracellular volume expansion are frequently encountered complications of congestive heart failure (HF) that can cause morbidity and mortality. Tolvaptan (Otsuka) is an orally administered nonpeptide vasopressin (VP) V2 receptor antagonist that inhibits water reabsorption in the kidney by competitively blocking VP binding, resulting in water diuresis without significantly changing total electrolyte excretion. In the 24-hour period following a 30-mg dose of tolvaptan, urine excretion rate increases and declines as plasma concentrations rise and fall; this uneven effect results in 80% of daily urine output in the first 12 hours. Therefore, the current study was designed to assess the pharmacodynamic effects, pharmacokinetics, and clinical safety of tolvaptan 30 mg QD plus placebo versus 15 mg BID over 7 days in patients with NYHA Class II/III heart failure and persistent fluid overload, SBP > or = 90 mm Hg, and a serum creatinine < or = 3.0 mg/dL. Patients were withdrawn from diuretics for 48 hours before randomization. Statistics were performed with ANCOVA for continuous variables and Mantel-Haenszel mean score test stratified by center for categorical variables. Thirty-nine of 40 patients completed days 1 and 7. There were no significant clinical, pharmacokinetic, or pharmacodynamic differences between the dosing regimens over time. Based on these findings, tolvaptan 30 mg was chosen as the comparator for placebo in a large phase 3 survival trial. PMID- 16220068 TI - Caveolin scaffolding peptide-1 interferes with norepinephrine-induced PLC-beta activation in cultured rat vascular smooth muscle cells. AB - Caveolins are a family of integral membrane proteins implicated in various cell functions, including the organization and inactivation of signaling molecules of G protein-coupled receptors. We tested the ability of human caveolin scaffolding peptide-1 (CSP-1) to regulate norepinephrine- (NE) or histamine (HIS)-induced increases on intracellular calcium concentrations ([Ca(2+)]i). In cultured rat vascular smooth muscle cells (VSMC), CSP-1 inhibited in a concentration-dependent manner NE- and HIS-induced increases in [Ca(2+)]i. This effect can be explained by the fact that CSP-1 inhibited a common signaling pathway. We tested the ability of this peptide to decrease the activation of PLC-beta3 and MAPK. CSP-1 inhibited the expression of the activated form of both enzymes, suggesting a direct effect of the peptide on the signaling cascade. CSP-1 readily enters VSMC in culture, as observed when FITC-conjugated CPS-1 is added to cell culture media. Taken together, these data suggest that CSP-1 blocks the effects of NE and HIS on [Ca(2+)]i of VSMC by inhibiting the activation of PLC-beta3 and MAPK. PMID- 16220069 TI - Coronary microvascular dysfunction in patients with microvascular angina: analysis by TIMI frame count. AB - We have previously reported that angina pectoris persists in patients with coronary microvascular spasm (MVS) even on calcium channel blockers. Because measurement of myocardial lactate production in the coronary sinus is necessary to diagnose MVS, a more feasible diagnostic method needs to be developed. In this study, we examined the diagnostic significance of Thrombolysis in Myocardial Infarction (TIMI) frame count, a marker of coronary blood flow, in 131 consecutive patients who underwent provocation test for coronary spasm with acetylcholine (ACh). Epicardial coronary spasm (ES) was diagnosed as more than 75% of ACh-induced vasoconstriction noted by coronary angiography. MVS was diagnosed as ACh-induced myocardial ischemia (chest pain, ischemic ECG changes, and myocardial lactate production) without ES. TIMI frame count was significantly increased in patients with MVS alone (n = 35) and those with ES + MVS (n = 16) compared with those with ES alone (n = 53) or those with no myocardial ischemia (Normal, n = 27) either before and after intracoronary ACh and even after intracoronary isosorbide dinitrate (ISDN) in both the left anterior descending (LAD) and the left circumflex coronary artery (LCX). TIMI frame count in LAD correlated well to that in LCX in patients with MVS, suggesting diffuse impaired coronary microcirculation in the myocardium. These results suggest that increased TIMI frame count in response to ACh reflects microvascular dysfunction in MVS and that ISDN may not be enough to relieve MVS. Thus, TIMI frame count may be useful to diagnose MVS without requiring coronary sinus catheterization or myocardial lactate production measurement. PMID- 16220070 TI - Effects of thiol chelation on alpha1-adrenoceptor-induced vasoconstriction in vivo. AB - The aims of this study were to determine whether systemic injections of the lipophobic thiol chelator, para-hydroxymercurobenzoic acid (PHMBA) would reduce the vasoconstrictor responses elicited by the alpha1-adrenoceptor agonist, phenylephrine, in urethane-anesthetized rats by chelation of thiol residues in alpha1-adrenoceptors in vascular smooth muscle rather than voltage-sensitive Ca(2+)-channels (Ca(2+)VERSUS-channels). The magnitudes and durations of the vasoconstrictor responses elicited by phenylephrine were markedly reduced after the injections of PHMBA. In contrast, the maximal phenylephrine-induced responses were not affected whereas the durations of these responses were markedly attenuated after injection of the Ca(2+)VERSUS-channel blocker, nifedipine. Nifedipine elicited pronounced and sustained falls in mean arterial blood pressure and vascular resistances in PHMBA-treated rats. Moreover, the vasodilator actions of the nitric oxide-donor, sodium nitroprusside were minimally attenuated by PHMBA whereas they were markedly attenuated by nifedipine. These findings support evidence that the vasoconstrictor responses due to activation of alpha1-adrenoceptors are initiated by mobilization of intracellular pools of Ca(2+) whereas they are sustained by opening of Ca(2+)VERSUS-channels. These findings also suggest that PHMBA diminishes the vasoconstrictor effects of phenylephrine by chelation of thiol residues in alpha1 adrenoceptors rather than by blockade of Ca(2+)VERSUS-channels, and that chelation of these thiol residues prevents agonist occupation and/or activation of these receptors and subsequent mobilization of intracellular pools of Ca(2+). PMID- 16220071 TI - Peroxynitrite elicits dysfunction of stereoselective s-nitrosocysteine recognition sites. AB - The aim of this study was to determine whether induction of tachyphylaxis to peroxynitrite (induced by giving 10 intravenous injections of a 10-micromol/kg dose) differentially affects the vasodilator responses elicited by systemic injections of the L- and D-isomers of S-nitrosocysteine (L-SNC and D-SNC), in pentobarbital-anesthetized rats. L- and D-SNC (12.5-200 nmol/kg, iv) elicited dose-dependent reductions in hindquarter, mesenteric, and renal vascular resistances. The L-SNC-induced vasodilator responses in the hindquarter and renal vascular beds were virtually abolished whereas the vasodilator responses in mesenteric bed were markedly diminished after administration of peroxynitrite. The D-SNC-induced vasodilator responses in the hindquarter and renal beds were slightly attenuated whereas the vasodilator responses in the mesenteric bed were not diminished after administration of peroxynitrite. The vasodilator responses elicited by the nitric oxide donor, MAHMA NONOate (5-50 nmol/kg, iv), were not attenuated by peroxynitrite. The finding that induction of tachyphylaxis to peroxynitrite diminishes the effects of L- and D-SNC but not MAHMA NONOate suggests that the stereoisomers exert their vasodilator effects by mechanisms other than their decomposition to nitric oxide. Moreover, the finding that induction of tachyphylaxis to peroxynitrite causes a more pronounced attenuation of the vasodilator effects of L- than D-SNC supports evidence that the stereoisomers differentially interact with stereoselective S-nitrosothiol recognition sites in the vasculature. Taken together, these novel results support the possibility that peroxynitrite diminishes the vasodilator potencies of L- and D-SNC by oxidation and/or nitration of amino acids in these recognition sites. PMID- 16220072 TI - Loss of K+ATP-channel-mediated vasodilation after induction of tachyphylaxis to peroxynitrite. AB - Systemic injections of peroxynitrite elicit pronounced vasodilator responses in rats by activation of ATP-dependent K+ channels (K+ATP-channels). The aim of this study was to determine whether development of tachyphylaxis to the vasodilator actions of peroxynitrite involves the loss of K+ATP-channel function. The falls in mean arterial blood pressure (MAP) and mesenteric and hindquarter vascular resistances produced by the K+ATP-channel agonist, cromakalim (3-18 microg/kg, iv), and the nitric oxide (NO) donor, sodium nitroprusside (SNP; 1-4 microg/kg, iv), were determined in pentobarbital-anesthetized rats before and after induction of tachyphylaxis to peroxynitrite induced by the administration of 10 injections of peroxynitrite (10 micromol/kg, iv). The first dose of peroxynitrite elicited pronounced falls in MAP and vascular resistances whereas the tenth injection elicited much smaller responses that were equivalent to those of decomposed peroxynitrite. Before induction of tachyphylaxis to peroxynitrite, cromakalim and SNP produced dose-dependent reductions in MAP and vascular resistances. The hemodynamic actions of cromakalim were markedly attenuated after induction of tachyphylaxis to peroxynitrite whereas the SNP-induced responses were only slightly attenuated. These results suggest that tachyphylaxis to the vasodilator actions of peroxynitrite involves the loss of K+ATP-channel function whereas tachyphylaxis to peroxynitrite minimally affects NO-mediated vasodilation. Taken together, these findings raise the possibility that peroxynitrite inhibits K+ATP-channel function by oxidation and/or nitration of amino acids in these channels. PMID- 16220073 TI - Role of ATP-sensitive K+ -channels in hemodynamic effects of peroxynitrite in anesthetized rats. AB - The aim of this study was to determine whether the hypotensive and vasodilator actions of peroxynitrite in pentobarbital-anesthetized rats involve the activation of ATP-sensitive K+-channels (K+ATP-channels). The effects of the K+ATP-channel agonist, cromakalim (9-36 microg/kg, iv), peroxynitrite (0.5-10 micromol/kg iv), and L-S-nitrosocysteine (12.5-200 nmol/kg, iv) on mean arterial blood pressure (MAP) and mesenteric (MR) and hindquarter (HQR) vascular resistances were determined before and after injection of the K+ATP-channel blocker, glibenclamide (40 micromol/kg, iv). Cromakalim, peroxynitrite, and L-S nitrosocysteine produced dose-dependent reductions in MAP, MR, and HQR. Administration of glibenclamide did not affect resting hemodynamic parameters but markedly attenuated the hemodynamic actions of cromakalim. The maximal falls in MAP and HQR produced by peroxynitrite were attenuated by glibenclamide whereas the maximal falls in MR were not affected. In addition, the duration of the hypotensive and vasodilator effects of peroxynitrite in the mesenteric and hindquarter beds were markedly diminished by glibenclamide. In contrast, glibenclamide did not affect the maximal hypotensive or vasodilator effects of L S-nitrosocysteine or the duration of these responses. These results suggest that the hypotensive and vasodilator actions of peroxynitrite in anesthetized rats involve the activation of K+ATP-channels whereas the hemodynamic actions of L-S nitrosocysteine do not. PMID- 16220074 TI - Differentiation of L- and D-S-nitrosothiol recognition sites in vivo. AB - The main aim of this study was to determine the effects of the lipophobic electron acceptor, nitroblue tetrazolium (NBT), on the vasodilator responses elicited by femoral vein injections of L- and D-S-nitrosocysteine (L- and D-SNC), L- and D-S-nitroso-beta,beta-dimethylcysteine (L- and D-SNPEN) and the nitric oxide (NO) donor, MAHMA NONOate, in pentobarbital-anesthetized rats. L- and D SNC, L- and D-SNPEN, and MAHMA NONOate elicited dose-dependent falls in mean arterial blood pressure (MAP), and hindquarter (HQR), renal (RR), and mesenteric (MR) vascular resistances. The L-SNC- and L-SNPEN-induced depressor and vasodilator responses were markedly attenuated after injection of NBT. The D-SNC- and D-SNPEN-induced falls in mean arterial pressure, hindquarter, and mesenteric vascular resistances were also reduced after injection of nitroblue tetrazolium whereas the falls in renal resistances were not affected. However, nitroblue tetrazolium inhibited the L-SNC and L-SNPEN responses much more profoundly than the D-SNC and D-SNPEN responses in each vascular bed. In contrast, the MAHMA NONOate-induced responses were not attenuated by nitroblue tetrazolium. This study demonstrates that nitroblue tetrazolium attenuates L- and D-SNC-and L- and D-SNPEN- mediated but not NO-mediated vasodilation. The lack of effects of NBT on the NO responses suggests that NBT does not interfere with the intracellular mechanisms by which NO relaxes vascular smooth muscle. The more pronounced effects of NBT on the vasodilator effects of L-SNC and L-SNPEN than D-SNC and D SNPEN suggests that these stereoisomers differentially interact with stereoselective S-nitrosothiol recognition sites in the vasculature and that these sites (or their signaling elements) contain thiol residues that may be susceptible to occupation and/or oxidation (ie, disulfide-bond formation) by nitroblue tetrazolium. PMID- 16220075 TI - Troglitazone inhibits long-term glycation and oxidation of low-density lipoprotein. AB - Troglitazone (T) is a member of a new class of antidiabetic drugs termed thiazolidinediones (TZDs), which has previously been used as an anti-diabetic agent. In this study we investigated the influence of T, a ligand for PPAR-gamma receptor, on copper-catalyzed or cell-mediated oxidation of native, glycated, and glycoxidated low-density lipoprotein (LDL). A dose-dependent inhibition of copper mediated low-density lipoprotein-oxidation, as monitored by the formation of oxidation-specific fluorescence, was observed for both native and glycated low density lipoprotein. At the concentration of 20 microg/mL the inhibition amounted from 14.7% to 64.7% by all low-density lipoprotein forms. For glycated low density lipoprotein we obtained the highest oxidation rate, but the most pronounced inhibition by T was found for glycoxidated low-density lipoprotein (goLDL). Inhibitory effects of T were also investigated by measurement of relative electrophoretic mobility (REM) in the concentration range of 0 to 20 microg/mL. The inhibition of 4h oxidation of native low-density lipoprotein was found in the entire concentration range, but significance was seen at 10 microg/mL. The long-term glycation and glycoxidation of low-density lipoprotein as measured by 5-hydroxymethyl-2-furaldehyde (5-HMF) formation and binding of fructosamine was found to be inhibited by T. In endothelial cell-mediated oxidation of low-density lipoprotein cytotoxicity of T in the concentration range of 0 to 160 microg/mL during 2 to 24 h oxidation was investigated. In the non cytotoxic concentration range of 5 to 20 microg/mL, a significantly reduced liberation of isoprostane 8-epi-PGF2alpha during 24 h cell-mediated oxidation of low-density lipoprotein and its modifications was found. This inhibitory action of T was most significant in the case of goLDL and amounted to approximately 20% to 60% inhibition at 5 to 20 microg/mL T, respectively. In the concentration range of 40 to 160 microg/mL, however, T showed an increasing cytotoxic action, as evidenced by loss of cell adhesion, loss of cellular protein, morphological changes, and cell disintegration as well as by strongly enhanced troglitazone mediated isoprostane 8-IP liberation (fivefold to sixfold). T may be used as a model to explore the thiazolidinediones' mechanism on oxidation in a more general aspect for treatment for T2DM, because T is not clinically available. PMID- 16220076 TI - Endothelin-1 inhibits inward rectifier K+ channels in rabbit coronary arterial smooth muscle cells through protein kinase C. AB - We studied inward rectifier K+ (Kir) channels in smooth muscle cells isolated from rabbit coronary arteries. In cells from small- (<100 microm, SCASMC) and medium-diameter (100 approximately 200 microm, MCASMC) coronary arteries, Kir currents were clearly identified (11.2 +/- 0.6 and 4.2 +/- 0.6 pA pF at -140 mV in SCASMC and MCASMC, respectively) that were inhibited by Ba(2+) (50 microm). By contrast, a very low Kir current density (1.6 +/- 0.4 pA pF) was detected in cells from large-diameter coronary arteries (>200 microm, LCASMC). The presence of Kir2.1 protein was confirmed in SCASMC in a Western blot assay. Endothelin-1 (ET-1) inhibited Kir currents in a dose-dependent manner. The inhibition of Kir currents by ET-1 was abolished by pretreatment with the protein kinase C (PKC) inhibitor staurosporine (100 nM) or GF 109203X (1 microm). The PKC activators phorbol 12,13-dibutyrate (PDBu) and 1-oleoyl-2-acetyl-sn-glycerol (OAG) reduced Kir currents. The ETA-receptor inhibitor BQ-123 prevented the ET-1-induced inhibition of Kir currents. The amplitudes of the ATP-dependent K+ (KATP), Ca(2+) activated K+ (BKCa), and voltage-dependent K+ (KV) currents, and effects of ET-1 on these channels did not differ between SCASMC and LCASMC. From these results, we conclude that Kir channels are expressed at a higher density in SCASMC than in larger arteries and that the Kir channel activity is negatively regulated by the stimulation of ETA-receptors via the PKC pathway. PMID- 16220077 TI - C-reactive protein augments interleukin-8 secretion in human peripheral blood monocytes. AB - C-reactive protein (CRP) is a powerful predictor and risk factor for cardiovascular diseases. The CXC- and CC-type chemokines interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) are important chemokines for leukocyte trafficking identified in atheromatous plaque expressed mainly by macrophages in humans. We assessed whether C-reactive protein could induce MCP-1 and IL-8 secretion. In human peripheral blood monocytes, C-reactive protein (12.5-50 microg/mL) increased IL-8, but not MCP-1 secretion in a time- (6-24 hours) and dose-dependent manner as detected by ELISA. C-reactive protein could augment the production of reactive oxygen species (ROS) as measured by chemiluminescence and inhibitors of NAD(P)H oxidase (DPI and PAO) and ROS scavengers (superoxide dismutase, catalase, and 1% dimethyl sulphoxide) abolished C-reactive protein induced IL-8 secretion. Furthermore, relative quantity of IL-8 mRNA was significantly increased by C-reactive protein 50 microg/mLfor 12 hours, which could be inhibited by DPI 1 microM or superoxide dismutase (SOD) 250 U/mL. The inhibitors of ERK 1/2 (PD98059), p38 (SB203580) MAPK, and NF-kappaB (PDTC and MG132) significantly decreased C-reactive protein-induced IL-8 secretion in human monocytes. Also, agonists of peroxisome proliferator-activated receptor (PPAR) alpha (WY14643) and PPARgamma (troglitazone) could largely inhibit C-reactive protein responses. Thus, our data indicate that C-reactive protein at pathologic levels increases IL-8 secretion and mRNA via enhancing ROS derived mainly from NAD(P)H oxidase and the subsequent activation of ERK1/2, p38 MAPK, and NF-kappaB. The activation of PPARalpha/gamma can negatively regulate C-reactive protein induced IL-8 production in human monocytes. PMID- 16220078 TI - Time-dependent changes in antioxidant enzymes and vascular reactivity of aorta in streptozotocin-induced diabetic rats treated with curcumin. AB - In the present study changes in oxidative stress and vascular reactivity in aortic rings of chronic streptozotocin-diabetic (STZ-CON) and nondiabetic (ND CON) rats is studied at 4 weekly intervals up to 24 weeks. The effect of chronic curcumin (200 mg/kg) treatment was also studied. Blood glucose and blood pressure levels were significantly higher in the STZ-CON group and curcumin administration had no significant effect on it. Superoxide dismutase and catalase activity were either unchanged or significantly increased during the early stage of diabetes whereas during the medium and late stage were significantly reduced. Reduced glutathione and lipid peroxidation levels significantly decreased as time after STZ administration increased. Phenylephrine (PE)-induced contraction was significantly (P < 0.05) increased during the early stage of diabetes, whereas it was significantly (P < 0.05) reduced at the medium and late stage of diabetes. Acetylcholine (Ach)-induced relaxation significantly decreased with respect to time after STZ administration. Sodium nitroprusside (SNP)-induced relaxation was unaltered up to initial stage but after medium stage there was a rightward shift and the pD2 value significantly decreased. Though curcumin treatment had no significant effect on superoxide dismutase, catalase, and reduced glutathione levels, it significantly reduced lipid peroxidation compared with diabetic control. Curcumin treatment attenuated the phenylephrine-induced increase in contraction during the early stage. However, curcumin treatment had no significant effect at the medium and late stage. Though curcumin administration improved Ach-induced relaxation it did not restore it to normal. Inability of curcumin to prevent oxidative stress during the late stage may be due to the fact that chronic diabetes (hyperglycemia) leads to excessive production of free radicals. Hence the present study shows that variations reported in antioxidant enzymes and vascular reactivity are due to the duration of diabetes or time after diabetes induction in STZ model and this can not be completely reversed by chronic treatment with curcumin. PMID- 16220079 TI - Chronic At1 receptor blockade alters the mechanisms mediating hypoxic dilation in middle cerebral arteries. AB - The purpose of this study was to determine whether chronic blockade of the angiotensin II (ANG II) AT1 receptor under normal physiological conditions impairs vascular relaxation mechanisms in isolated middle cerebral arteries (MCA). Male Sprague-Dawley rats on a standard diet were given losartan (1 mg/mL) in the drinking water or normal water ad libitum for 7 days. Vessel diameters were measured by television microscopy before and during exposure to various vasodilator agonists and reductions in PO2 from 140 mm Hg to 35-45 mm Hg. Dilations to acetylcholine (1 microM), the stable prostacyclin analogue iloprost (10 pg/mL), and the Gs protein activator cholera toxin (1 ng/mL) were completely eliminated in vessels from losartan-treated animals. However, middle cerebral arteries from control and losartan-treated rats still demonstrated significant dilations in response to reduced PO2. Hypoxic dilation of middle cerebral arteries from control rats was eliminated by indomethacin (1 microM) and unaffected by the NOS inhibitor L-NAME (100 microM) whereas dilation in response to reduced PO2 in middle cerebral arteries from losartan-treated rats was eliminated by L-NAME and unaffected by indomethacin. Middle cerebral arteries from control and losartan-treated animals exhibited similar dilations in response to the NO-donor sodium nitroprusside (1 microM). These data suggest that AT1 receptor activation is important in maintaining normal vascular relaxation mechanisms in cerebral resistance arteries during normal physiological conditions, and that AT1 receptor blockade causes a shift in the mechanisms of hypoxic dilation of middle cerebral arteries from cyclooxygenase metabolites to NO. PMID- 16220081 TI - The Pediatric Infectious Diseases Society Annual Awards, 2005. PMID- 16220080 TI - Influence of CYP2D6 genotype on metoprolol plasma concentration and beta adrenergic inhibition during long-term treatment: a comparison with bisoprolol. AB - In patients routinely treated with metoprolol, influences of CYP2D6 genotype on the response of heart rate to isoproterenol (IP) were studied at its peak and trough concentrations and were compared with those of bisoprolol. In 72 patients treated with metoprolol or bisoprolol, CYP2D6 genotype (ie, CYP2D6*1, *2, *4, *5, *10, and *14) was determined. No patients except one who was heterozygous for CYP2D6*5 carried the null alleles of CYP2D6. The homozygote frequency for CYP2D6*10 was relatively high (19.4%) and these patients had greater peak and trough plasma concentrations of metoprolol than the other patients. Isoproterenol induced percentage increases in heart rate were 58% and 38% less at the low and high rate of isoproterenol infusion (0.02 and 0.04 microg/kg/min), respectively, in patients homozygous for CYP2D6*10 than in the other patients at the trough, but not at the peak concentrations. In contrast, CYP2D6 genotype did not affect plasma concentrations of bisoprolol and the extent of its beta-adrenergic inhibition. Thus, in patients routinely treated with metoprolol, CYP2D6 genotype significantly affects circadian variations of beta-adrenergic inhibition induced by metoprolol. In contrast, bisoprolol has a relatively constant beta-adrenergic inhibition independent of CYP2D6 genotype. PMID- 16220082 TI - Multicenter study to assess safety and efficacy of INH-A21, a donor-selected human staphylococcal immunoglobulin, for prevention of nosocomial infections in very low birth weight infants. AB - BACKGROUND: Prophylactic administration of intravenous immunoglobulin has been inconsistent in reducing the risk of sepsis in very low birth weight (VLBW) infants presumably because of varying titers of organism specific IgG antibodies. INH-A21 is an intravenous immunoglobulin from donors with high titers of antistaphylococcal antibodies. This dose-ranging study explored safety and preliminary activity of INH-A21 for prevention of staphylococcal sepsis in VLBW infants. METHODS: This was a multicenter, double blind, group-sequential study. Infants with birth weights 500-1250 g were randomized to receive up to 4 doses of placebo, 250 mg/kg, 500 mg/kg or 750 mg/kg INH-A21. Safety and frequencies of sepsis were compared across treatment groups. RESULTS: All treatment groups had similar mean gestational age, birth weight, Apgar score and maternal use of antibiotics. Randomizations to 250 mg/kg (N = 94) and 500 mg/kg (N = 96) doses were terminated after interim analyses demonstrated a low probability of finding a difference when compared with placebo. Infants randomized to the INH-A21 750 mg/kg group (N = 157) had fewer episodes of Staphylococcus aureus sepsis [relative risk (RR), 0.37; 95% confidence interval (CI), 0.12-1.12; P = 0.14], candidemia (RR 0.34; 95% CI 0.09-1.22; P = 0.09) and mortality (RR 0.64; 95% CI 0.25-1.61; P = 0.27) when compared with the placebo-treated cohort (N = 158). No dose-related trends were observed for adverse events or morbidities associated with prematurity. CONCLUSIONS: INH-A21 750 mg/kg demonstrated potential to reduce sepsis caused by S. aureus, candidemia and mortality in VLBW infants. Although statistical significance was not reached, based on the magnitude of the estimated differences, the efficacy and safety of INH-A21 750 mg/kg should be evaluated in an adequately powered, well-controlled study. PMID- 16220083 TI - Safety and antiviral response at 12 months of lopinavir/ritonavir therapy in human immunodeficiency virus-1-infected children experienced with three classes of antiretrovirals. AB - INTRODUCTION: Many human immunodeficiency virus type 1 (HIV-1)-infected children have already failed treatment with 2 or even 3 classes of antiretrovirals. Coformulation of lopinavir with low dose ritonavir exhibits a potent antiretroviral effect. However, the data in heavily pretreated children are still scarce. This study evaluated the safety and effectiveness of combination therapy including lopinavir/ritonavir in children with prior exposure to all classes of oral antiretrovirals. METHODS: This was an open label multicenter observational study, in which data were reviewed according to a standardized protocol. The study population included all HIV-1-infected children with virologic failure (HIV 1 RNA >5000 copies/mL) followed in 12 Spanish hospitals for >12 months, experienced with the 3 classes of oral antiretrovirals, in whom a lopinavir/ritonavir-containing regimen was started. RESULTS: By March 2003, 45 patients had been treated with lopinavir/ritonavir for a median of 18 months (range, 3-28). The median age at baseline was 9.7 years (range, 4.3-17.1). The median times of prior treatment were 88 months (range, 31-145) with nucleoside reverse transcription inhibitors and 42 months (range, 19-63) with protease inhibitors. Twenty-five patients were classified as Centers for Disease Control and Prevention clinical category C. Median values for absolute and percentage CD4 at baseline were 501 (range, 6-1512) and 19% (range, 0.5-49), respectively, and plasma HIV-RNA was 5.0 log10 copies/mL (range, 4.1-6.1). During follow-up, 11 (24%) children switched from liquid to solid formulation. At 48 weeks, the median values for absolute and percentage CD4 increased by 199 cells/microL and 3%, respectively, and median plasma viral load declined 1.75 log10 copies/mL. Forty two percent of children achieved a plasma RNA of <400 copies/mL (intent to treat analysis). Baseline genotypic resistance was available in 40 children. Nonresponders had 7.0 +/- 1.6 protease inhibitor-associated mutations at baseline compared with 4.8 +/- 1.7 in children achieving virologic suppression (P = 0.06). Adverse events were described in 18 children. Three children permanently discontinued and 4 transiently withdrew lopinavir/ritonavir. At 12 months, there were mild but not significant increases in plasma cholesterol and triglycerides. CONCLUSIONS: Lopinavir/ritonavir when given as part of salvage regimen is well tolerated, although switching to pills is frequently required. The regimen has a potent and durable antiretroviral activity in most heavily pretreated children, despite the presence of multiple mutations to all classes of oral antiretrovirals. PMID- 16220084 TI - Pharmacokinetics and 24-week efficacy/safety of dual boosted saquinavir/lopinavir/ritonavir in nucleoside-pretreated children. AB - OBJECTIVE: To assess the pharmacokinetics and 24-week efficacy and safety of dual boosted saquinavir/lopinavir/ritonavir combination in children. DESIGN: Twenty reverse transcription inhibitor-pretreated children at 2 centers in Thailand were treated with saquinavir/lopinavir/ritonavir in an open label, single arm, 6-month prospective study. The dosage was 50 mg/kg twice daily (bid) for saquinavir and 230/57.5 mg/m bid for lopinavir/ritonavir. Ten children also received lamivudine. METHODS: Samples were collected for a 12-hour pharmacokinetic profile in all children. Plasma concentrations of saquinavir, lopinavir and ritonavir were determined using a validated high performance liquid chromatography technique. RESULTS: At baseline, the median age was 8.5 years, with human immunodeficiency virus (HIV) RNA 4.9 log10 copies/mL, CD4 count 129 cells/microL and CD4%, 6.5%. Median area under the concentration curve at 0-12 hours and Cmin were 39.4 mg/L.h and 1.4 mg/L for saquinavir and 118 mg/L.hr and 5.9 mg/L for lopinavir. After 24 weeks of treatment, HIV RNA was suppressed below 400 copies/mL for 16 of 20 (80%) children (intent-to-treat analysis) and below 50 copies/mL for 12 of 20 children (60%), and CD4% (count) rose by a median of 6% (216 cells/microL). Median changes of triglyceride and total cholesterol were 56 and 36.5 mg/dL, respectively (P = 0.01). Lopinavir Cmin <1 and saquinavir Cmin <0.28 mg/L correlated with HIV RNA >400 copies/mL, and lopinavir Cmax >15 mg/L correlated with rises in cholesterol (P < 0.05). CONCLUSION: Plasma drug concentrations of saquinavir, lopinavir and ritonavir were at the higher limits of expected ranges for adult treatment at approved dosages (1000/100 mg bid for saquinavir, 400/100 mg bid for lopinavir/ritonavir). The regimen was well-tolerated and had good efficacy at 24 weeks. This dual boosted protease inhibitor combination should be assessed in larger trials of reverse transcription inhibitor-experienced children. PMID- 16220085 TI - Efficacy, tolerability and pharmacokinetics of two nelfinavir-based regimens in human immunodeficiency virus-infected children and adolescents: pediatric AIDS clinical trials group protocol 403. AB - INTRODUCTION: Few combinations of highly active antiretrovirals have been studied in nucleoside reverse transcription inhibitor (NRTI)-experienced, human immunodeficiency virus (HIV)-infected children. We tested the efficacy, tolerability and pharmacokinetics of 2 combination therapies containing an NRTI, protease inhibitors +/- a nonnucleoside reverse transcription inhibitor (NNRTI). METHODS: This was a phase II, randomized, multicenter study. Forty-one children and youths between 5 months and 21 years with prior NRTI and no prior NNRTI or protease inhibitor experience received either nelfinavir (NFV) 30 mg/kg twice daily (bid), ritonavir (RTV) 400 mg/m bid and buffered didanosine (ddI) 240 mg/m daily (arm A) or NFV 50-55 mg/kg bid, nevirapine (NVP) 120 mg/m bid and stavudine (d4T) 1 mg/kg bid (arm B). Patients were evaluated clinically for 48 weeks after initiation of therapy. Intensive pharmacokinetic sampling occurred after 4 weeks of therapy. RESULTS: : The proportion of children with HIV-1 RNA < or =400 copies/mL and on randomized treatment at 48 weeks was 65% among children assigned NFV + RTV + ddI versus 28% among those assigned NFV + NVP + d4T (P = 0.039). No significant difference in median CD4% change from baseline to week 48 was found (3% versus 1%). No significant differences in safety or tolerability between children randomized to NFV + RTV + ddI versus NFV + NVP + d4T were identified. However, a trend toward a higher rate of permanent discontinuation of study treatment was noted among children assigned to NFV + NVP + d4T compared with NFV + RTV + ddI [7 of 20 (35%) versus 2 of 21 (10%); P = 0.12]. NFV pharmacokinetic measurements were not statistically different between the treatment groups, yet exposure to the NFV metabolite, M8, was significantly higher in subjects receiving RTV. The pharmacokinetics for NVP, RTV and d4T were similar to those of previously reported data. CONCLUSION: : Combination therapy containing NFV + RTV + ddI appears more efficacious in NRTI-experienced children than a regimen containing NFV + NVP + d4T. Differences in tolerability between the 2 treatment groups were not identified. Systemic exposure of these drugs was similar to that reported in other HIV-infected children and adults. PMID- 16220086 TI - Pharmacokinetics of terbinafine in young children treated for tinea capitis. AB - BACKGROUND: Dermatophytes are the most common cause of human fungal infections. Response rates to existing therapy are lower than optimal, but newer agents like terbinafine hold promise for improved management of such infections. This investigation was designed to evaluate the single dose and steady state pharmacokinetics of terbinafine in young children with tinea capitis. METHODS: Twenty-two otherwise healthy children (4-8 years) with tinea capitis were eligible for enrollment. Children were treated with terbinafine once daily according to body weight (<25 kg, 125 mg; 25-35 kg, 187.5 mg), and pharmacokinetic sampling was conducted after the first dose, at the midpoint of treatment and at steady state. Plasma terbinafine concentrations were quantitated, and the pharmacokinetic indices compared with adult data. RESULTS: Absolute estimates of Cmax and area under the concentration curve (AUC)0-24 were comparable between children and adults for the administered dose; however, children demonstrated significantly lower estimates of exposure when dose was corrected for weight (Cmax SS 200 +/- 104 versus 454 +/- 185 ng/mL per mg/kg dose, P < 0.01; AUCSS: 1110 +/- 640 versus 2756 +/- 1775 ng*h/mL per mg/kg dose, P < 0.01). When examined along a continuum, age accounted for approximately 50% of the variability observed in dose-normalized Cmax and AUC (P < 0.01). A slight but significant reduction in apparent oral clearance was observed with increasing age (0.02 L/h/kg per yr) that likely accounts for the lesser degree of accumulation observed in children at steady state (accumulation ratio, 1.5 +/- 0.8 versus 2.3 +/- 0.6, P < 0.01). Adverse events consisted principally of headache (n = 3) and gastrointestinal complaints (altered eating habits n = 3, loss of appetite n = 3, stomachache n = 4, diarrhea n = 2). A reduction in neutrophil count was observed in 5 children and thought to be related to study drug in 2. CONCLUSIONS: Children require significantly larger weight-normalized doses to approximate the exposure estimates observed in adults. The dosing scheme used in this investigation results in absolute exposure estimates at steady state and a safety profile that are not appreciably different from adults. PMID- 16220087 TI - Tick-borne encephalitis in children in Styria and Slovenia between 1980 and 2003. AB - BACKGROUND: To describe the effect of the Austrian vaccination program against tick-borne encephalitis (TBE) on the incidence of this disease in children from Styria, an Austrian federal state, and to compare it with that in Slovenia, the neighboring country with a risk to acquire TBE similar to that of Styria. METHODS: A retrospective population-based cohort study was performed with the use of discharge data from all Styrian pediatric hospitals and data from the Center for Communicable Diseases at the National Institute of Public Health in Ljubljana, Slovenia. RESULTS: From January 1980 to December 2003, 139 cases of TBE in children younger than 16 years were observed in Styria. The annual incidence of TBE/100,000 Styrian children declined from 2.5-9.3 cases between 1980 and 1986 to 0-2.2 between 1987 and 1993 and to 0-1 between 1994 and 2003. Extrapolating the incidence of 6.3 cases/100,000 children between 1980 and 1986 to the time from 1994 to 2003, 124 pediatric TBE cases had been prevented in Styria in the past 10 years. CONCLUSIONS: Our data show that the Austrian vaccination program against TBE can lead to the nearly complete disappearance of TBE in children living in areas highly endemic for TBE. PMID- 16220088 TI - Acute trichinellosis in children compared with adults. AB - OBJECTIVES: Trichinellosis is a cosmopolitan parasite infection caused by Trichinella nematodes that is acquired from consumption of raw meat from several animal species. Knowledge of the clinical pattern and laboratory features of the disease in childhood is limited. The purpose is to study the clinical pattern of trichinellosis caused by Trichinella britovi in children and to compare it in household adults. METHODS: We evaluated all children up to 17 years of age and their adult householders exposed to the consumption of infected meat during an outbreak of trichinellosis. A questionnaire was developed to record clinical data. The blood sample was collected for blood count, muscle enzymes, serum electrolytes, albumin and serology. All exposed children were treated with mebendazole, and severe symptomatic patients received prednisolone. Clinical and laboratory presentations and outcome were recorded. To evaluate the clinical picture of trichinellosis in childhood, clinical and laboratory findings were compared between children and household adults with a confirmed diagnosis who consumed the same amount of infected meat. RESULTS: In 47 (62%) of 76 children with suspected trichinellosis, the diagnosis was serologically confirmed. The main clinical and laboratory findings in children were fever, abdominal pain, myalgia, facial and/or eyelid edema, rash, eosinophilia and increased muscular enzymes. The incubation period was similar in children and adults, but myalgia (66% versus 96%, P < 0.01), facial and/or eyelid edema (57% versus 86%, P < 0.05), eosinophilia (52% versus 96%, P < 0.01) and increased serum creatine kinase (38% versus 79%, P < 0.01) were less common in children than in adults. Seroconversion occurred in fewer children than adults, but the difference was not statistically significant. CONCLUSIONS: T. britovi infection shows a benign course and a milder clinical picture in children than in adults who consumed the same amount of infected meat. PMID- 16220089 TI - Clinical and epidemiologic characteristics of viral infections in a neonatal intensive care unit during a 12-year period. AB - BACKGROUND: The incidence of viral infections in patients treated in the neonatal intensive care unit (NICU) is not well-known. We summarized the data of all patients with laboratory-confirmed viral infections admitted at the NICU of our hospital during the period of 1992-2003. OBJECTIVES: To determine the incidence of viral infections among infants hospitalized in a NICU, the associated clinical manifestations and their outcome. METHODS: Retrospective analysis of epidemiologic, virologic and clinical data from infants with proven viral infection. The diagnosis viral infection was confirmed by positive viral culture and/or polymerase chain reaction from clinical samples. RESULTS: Viral infection was confirmed in 51 of 5396 infants (1%) admitted to the NICU; 20 (39%) had an enterovirus and parechovirus (EV/PEV) infection, 15 (29%) a respiratory syncytial virus (RSV) infection, 5 (10%) a rotavirus infection, 3 (6%) a cytomegalovirus (CMV) infection, 2 (4%) an adenovirus infection, 2 (4%) a parainfluenza virus infection, 2 (4%) a herpes simplex virus infection, 1 (2%) a rhinovirus infection and 1 (2%) a rubella virus infection. Three of the infants presented at birth with symptomatic rubella virus, CMV or herpes simplex virus infection. RSV infection developed mostly in hospitalized infants (60%), and 93% of infections occurred during the winter (November-March). The clinical presentations of EV/PEV disease were sepsis-like illness, prolonged seizures in term infants and gastrointestinal disease in preterm infants. RSV, parainfluenza virus, rhinovirus and CMV caused respiratory disease, predominantly in preterm infants. Gastrointestinal disease was seen only in preterm infants with adenovirus, rotavirus or EV/PEV infection. Mortality and serious sequelae were high in patients infected with EV/PEV (10 and 15%, respectively). CONCLUSIONS: The incidence of viral infection in the NICU was 1%. Enteroviral infections were the most frequently diagnosed infections, occurred often in term infants born at home and presented with sepsis-like illness or seizures. Preterm infants hospitalized from birth mainly developed gastrointestinal disease caused by rotavirus and adenovirus infection or respiratory disease caused by RSV, parainfluenza and CMV infection. Enteroviruses were responsible for the highest mortality and development of serious sequelae. PMID- 16220090 TI - Prevalence of human metapneumovirus and influenza virus infections among Japanese children during two successive winters. AB - BACKGROUND: Human metapneumoviruses (hMPVs) are recognized as a leading cause of respiratory infections in young infants in many countries. The objective of this study was to identify links between hMPV and influenza virus infections among children with influenza-like illness. METHOD: This study was conducted in 2 influenza seasons (2002-2003 and 2003-2004) at 2 pediatric outpatient clinics in Niigata city, Japan. Nasopharyngeal swabs or aspirates were collected from influenza-like illness patients, and hMPV and influenza were detected by reverse transcription-polymerase chain reaction (RT-PCR). A nucleotide sequence of 352 nucleotides segment of the F gene was performed. RESULTS: A total of 765 influenza viruses and 84 hMPV were identified from 1498 nasopharyngeal swabs or aspirates by virus isolation and RT-PCR, respectively. hMPV-positive rates in patients with influenza-like illness were 5.7 and 5.2% in the 2002-2003 and 2003 2004 seasons, respectively. Epidemic curves of influenza and hMPV patients showed similar patterns with peaks in February in 2 influenza seasons. hMPV infections occurred frequently in infants and school children. Approximately 46% of hMPV patients were coinfected with influenza A viruses, but those coinfected cases were not clinically distinct from the others. No coinfection with influenza B viruses was found. Phylogenetic analysis of the hMPV fusion gene sequences revealed that 2 distinct hMPV cocirculated and that completely identical strains in subgroup A were observed over 2 years. CONCLUSIONS: HMPV plays an important pathogenic role in patients with influenza-like illness in winter seasons, often in coinfections with influenza A viruses. PMID- 16220092 TI - Antimicrobial update: Daptomycin. PMID- 16220091 TI - Metaanalysis of short course antibiotic treatment for group a streptococcal tonsillopharyngitis. AB - OBJECTIVE: To compare bacterial and clinical cure rates in patients with group A streptococcal (GAS) tonsillopharyngitis treated with oral beta-lactam or macrolide antibiotics for 4-5 days versus 10-day comparators. METHODS: Medline, Embase, reference lists and abstract searches were used to identify available publications. Trials were included if there was bacteriologic confirmation of GAS tonsillopharyngitis, random assignment to antibiotic therapy for a beta-lactam or macrolide antibiotic of a shortened course versus a 10-day comparator and assessment of bacteriologic outcome using a throat culture. RESULTS: Twenty-two trials involving 7470 patients were included in 4 separate analyses. Trials were grouped by a short course of cephalosporins (n = 14), macrolides (other than azithromycin) (n = 6) and penicillin (n = 2). Cephalosporin trials were further grouped by the comparator, penicillin or the same cephalosporin. Short course cephalosporin treatment was superior for bacterial cure rate compared with 10 days of penicillin [odds ratio (OR), 1.47; 95% confidence interval (CI), 1.06 2.03]. For trials with short course macrolide therapy, OR = 0.79 (95% CI 0.59 1.06) neither the macrolides nor the 10-day comparators. Short course penicillin therapy was inferior in achieving bacterial cure versus 10 days of penicillin, OR = 0.29 (95% CI 0.13-0.63). Clinical cure rates mirrored bacteriologic cure rates. CONCLUSION: Superior cure rates can be achieved with shortened courses of cephalosporin therapy, but 5 days is inferior to 10 days of penicillin treatment. PMID- 16220094 TI - Hospital-based epidemiologic survey of malignancies in children infected with human immunodeficiency virus in Thailand. AB - To determine the incidence and spectrum of malignancies in human immunodeficiency virus-infected children, we surveyed 48 hospitals in Thailand between 1996 and 2000. There were 23 children (14 boys and 9 girls; average age at diagnosis of malignancy, 4.2 years), and the incidence rate was 0.6 per 1000 person-years. The most common malignancy was lymphoma (87.0%). The prognosis was poor. PMID- 16220093 TI - Advances in antifungal therapy. PMID- 16220095 TI - Immunogenicity and safety of inactivated influenza virus vaccine in young children in 2003-2004. AB - This study assessed the safety and immunogenicity of a pediatric formulation of the 2003-2004 inactivated, trivalent, split virion influenza vaccine, administered in a 2-dose schedule in healthy children ages 6-36 months, of whom 94% had protective titers (> or =1/40) to at least 1 antigen. The 2003-2004 split virion influenza vaccine was safe and immunogenic in young children. PMID- 16220096 TI - Detection of Kawasaki disease-associated antigen in inflamed gastrointestinal tract in acute Kawasaki disease. AB - We performed immunohistochemistry experiments using synthetic antibodies on the gastrointestinal tract and kidney from acute Kawasaki disease patients. Significant gastrointestinal tract inflammation was present in only 2/7 patients, who had antigen detected at the site of inflammation. Antigen was not detected in kidney from 6 patients. The presence of antigen may be related to gastrointestinal pathology in acute Kawasaki disease. PMID- 16220097 TI - Amebiasis in infancy in the middle-high socioeconomic class in Istanbul, Turkey. AB - Amebiasis is a major cause of morbidity and mortality worldwide. We present 11 children with acute Entamoeba histolytica infection. None of the infants developed extraintestinal disease or required hospitalization. Light microscopy results correlated with stool antigen test for E. histolytica. Breast-feeding and middle-high socioeconomic status does not protect infants from infection with E. histolytica in endemic areas. PMID- 16220098 TI - Viral shedding in children with influenza virus infections treated with neuraminidase inhibitors. AB - We examined the efficacy of neuraminidase inhibitors for reducing the duration of virus shedding after naturally occurring influenza virus infection. The duration of fever was significantly shorter in patients treated with neuraminidase inhibitors than in untreated patients. The durations of virus shedding from patients treated with neuraminidase inhibitors were not significantly shorter than those of untreated patients. PMID- 16220099 TI - West Nile virus encephalitis with thalamic involvement in an immunocompromised child. AB - West Nile virus has been an increasingly important pathogen in the United States since it was first reported in 1999. Neuroinvasive West Nile virus has been infrequently reported in the pediatric population. We report a case of severe West Nile virus encephalitis with cranial magnetic resonance imaging findings not yet described in children. PMID- 16220100 TI - Tremors and chorea induced by trimethoprim-sulfamethoxazole in a child with Pneumocystis pneumonia. AB - A 1.5-year-old girl developed high frequency tremors and chorea after receiving a dose of 120 mg/kg/d trimethoprim-sulfamethoxazole (TMP-SMX) for the treatment of Pneumocystis pneumonia. The child was human immunodeficiency virus-negative but immunocompromised because of prolonged immunosuppressive therapy. These symptoms disappeared 3 days after TMP-SMX was discontinued. Pediatricians should be aware of tremors and chorea among the potential adverse effects of high doses of TMP SMX. PMID- 16220101 TI - Invasive Cryptococcus laurentii disease in a nine-year-old boy with X-linked hyper-immunoglobulin M syndrome. AB - We report a 9-year-old boy with X-linked hyper-immunoglobulin M syndrome and invasive Cryptococcus laurentii infection identified on the basis of morphologic and biochemical characteristics and serologic tests. Fluconazole treatment resulted in complete recovery. We believe that C. laurentii should be included in the differential diagnosis of opportunistic fungal infections in patients with primary T cell immunodeficiencies. PMID- 16220103 TI - Invasive Exserohilum sinusitis in a patient with aplastic anemia. AB - A case of acute, invasive, sinusitis caused by the dematiaceous mold Exserohilum rostratum is presented. Herein we examine the subtypes of fungal sinusitis and briefly discuss some characteristics, pathologic and microbiologic differential diagnoses and management strategies of Exserohilum sinus infection in an immunocompromised patient. PMID- 16220102 TI - Superficial skin infection caused by Streptococcus pneumoniae in children. AB - Pneumococcal superficial skin infections are rarely described. We present 3 cases of Streptococcus pneumoniae superficial skin infections in previously healthy children. In 2 cases, lesions occurred on facial skin; in the third case they occurred on the scrotal raphe. One isolate was fully penicillin-resistant. PMID- 16220104 TI - Pseudomonas necrotizing fasciitis in an immunocompetent infant. PMID- 16220105 TI - Association of hyperimmunoglobulin E syndrome and Gianotti-Crosti syndrome. PMID- 16220106 TI - Vancomycin-resistant enterococcal bacteremia and necrotizing enterocolitis in a preterm neonate. PMID- 16220107 TI - Barriers to antibiotic lock therapy in children with intravascular catheter related bloodstream infections. PMID- 16220108 TI - ADH4 gene variation is associated with alcohol and drug dependence: results from family controlled and population-structured association studies. AB - We found strong associations between ADH4 gene variation and alcohol and drug dependence by the Hardy-Weinberg Disequilibrium (HWD) test and case-control association analysis in an initial study. The present study aimed to confirm these findings by controlling for population stratification and admixture effects to which the HWD test and case-control association methods may be vulnerable. In addition to 365 unrelated healthy controls and 560 unrelated cases in the initial study, we evaluated 104 small nuclear families with affected offspring who had diagnoses of alcohol and/or drug dependence. Seven single nucleotide polymorphisms (SNPs) at the ADH4 gene locus were genotyped in all subjects, and 38 unlinked ancestry-informative markers were also genotyped in unrelated cases and controls. Structured association analysis demonstrated that the genotypes of six ADH4 markers were associated with alcohol dependence, and all seven ADH4 markers were associated with drug dependence (P=10-0.047). Logistic regression analysis showed that: (i) the genotypes of SNP2 (rs1042363) were significantly associated with alcohol dependence and drug dependence (mainly cocaine dependence), and the genotypes of SNP3 (rs1126671) were also significantly associated with alcohol dependence and (ii) one seven-variant haplotype and one diplotype were significantly associated with alcohol dependence and other seven variant diplotypes were significantly associated with drug dependence (including cocaine and opioid dependence). Transmission disequilibrium test, haplotype-based haplotype relative risk and genotype-based haplotype relative risk analyses all confirmed the association of the ADH4 markers with alcohol dependence and drug dependence. Using rigorous study designs that account for possible population stratification, these findings confirm and extend our original observations indicating that variation at ADH4 predisposes to alcohol and drug dependence. PMID- 16220109 TI - Genotype-phenotype correlation between the polymorphic UGT2B17 gene deletion and NNAL glucuronidation activities in human liver microsomes. AB - The nicotine-derived tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK), is one of the most potent and abundant procarcinogens found in tobacco and tobacco smoke, and glucuronidation of its major metabolite, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), is an important mechanism for NNK detoxification. In cigarette smokers and tobacco chewers, there is a wide variation in the urinary levels of the ratio of NNAL to NNAL glucuronide (NNAL Gluc). To determine whether genetic variation plays a potential role in this inter-individual variability, NNAL-glucuronidating activities were analysed in a series of human liver microsomal specimens and compared with UGT2B17 deletion genotypes in the same subjects. Assays performed in vitro demonstrated that over expressed UGT2B17 exhibits high O-glucuronidating activity against NNAL. When stratifying subjects by UGT2B17 genotype, a significant or near-significant decrease in NNAL-O-Gluc formation was observed in liver microsomes from individuals who were either heterozygous [(+/0), P=0.07] or homozygous [(0/0), P=0.016] for the UGT2B17 deletion compared to liver microsomes from individuals with intact UGT2B17 alleles [(+/+)]. There was a significant (P<0.01) association between the level of liver microsomal NNAL-O-glucuronide formation and increasing numbers of the UGT2B17 null alleles in the liver microsomal specimens examined in this study, and a significant decrease in NNAL-O-Gluc formation was observed when comparing liver microsomes from individuals who had at least one UGT2B17 allele deleted [(+/0)+(0/0)] versus microsomes from UGT2B17 (+/+) subjects (P=0.004). When stratifying by the median value of NNAL-O-Gluc formation activity, a significantly (P=0.015) higher number of subjects with liver microsomes having low NNAL-O-Gluc formation activity contained the UGT2B17 null genotype compared to subjects with liver microsomes exhibiting high NNAL-O-Gluc formation activity. When stratifying by UGT2B7/UGT2B17 haplotypes, the association between the level of liver microsomal NNAL-O-glucuronide formation and increasing numbers of the UGT2B17 null allele was at the level of statistical significance for subjects with the UGT2B7 (*1/*2) (P=0.05) or UGT2B7 (*2/*2) (P<0.02) genotypes. These data suggest that the UGT2B17 deletion polymorphism is associated with a reduced rate of NNAL detoxification in vivo and may increase individual susceptibility to tobacco-related cancers. PMID- 16220110 TI - CYP2C9, CYP2C19, ABCB1 (MDR1) genetic polymorphisms and phenytoin metabolism in a Black Beninese population. AB - The genetically polymorphic cytochrome P450 2C9 (CYP2C9) metabolizes many important drugs. Among them, phenytoin has been used as a probe to determine CYP2C9 phenotype by measuring the urinary excretion of its major metabolite, S enantiomer of 5-(4-hydroxyphenyl)-5-phenylhydantoin (p-HPPH). Phenytoin pharmacokinetic is also dependent on the activity of CYP2C19 and p-glycoprotein (ABCB1). To determine the influence of CYP2C9, CYP2C19 and ABCB1 genetic polymorphisms on phenytoin metabolism in a Black population, 109 healthy Beninese subjects received a single 300 mg oral dose of phenytoin. Blood was drawn 4 h after drug intake and urine was collected during the first 8 h. Plasma phenytoin and urine S- and R-enantiomers of p-HPPH were determined by high-performance liquid chromatography. Urinary excretion of (S)-p-HPPH [defined as urinary volumex(S)-p-HPPH urinary concentration] and PMR (defined as the ratio of p-HPPH in urine to 4 h phenytoin plasma concentration), both markers of CYP2C9 activity, were used to determine the functional relevance of new variants of CYP2C9 (*5, *6, *8, *9 and *11) in this population. Plasma phenytoin concentration was significantly associated with ABCB1 haplotype/genotype (P=0.05, Kruskal-Wallis test) and levels increased significantly in the genotype order: wild-type, T3421A and Block-2 genotypes (P=0.015, Jonckheere-Terpstra test). Urinary excretion of (S)-p-HPPH and PMR were significantly associated with the CYP2C9 genotype (P=0.001, analysis of variance (ANOVA) and P<0.0001, Kruskal-Wallis test, respectively) and decreased in the order: CYP2C9*1/*1, CYP2C9*1/*9, CYP2C9*9/*9, CYP2C9*1/*8, CYP2C9*8/*9, CYP2C9*9/*11, CYP2C9*1/*5, CYP2C9*6/*9, CYP2C9*1/*6, CYP2C9*8/*11, CYP2C9*5/*8 and CYP2C9*5/*6 (P<0.001, Jonckheere-Terpstra test). A combined analysis of CYP2C9, 2C19 and ABCB1 revealed that only ABCB1 predicted phenytoin concentration at 4 h and explained 8% of the variability (r=0.08, P=0.04). On the other hand, only CYP2C9 was predictive for the urinary excretion of (S)-p-HPPH and PMR (r=0.21, P=0.001 and r=0.25, P<0.001, respectively). Furthermore, significant relation was found between urinary excretion of (R)-p HPPH and CYP2C9 genotype (P=0.035) and levels significantly increased in the genotype order: CYP2C9*1/*9, CYP2C9*1/*1, CYP2C9*9/*11, CYP2C9*1/*8 and CYP2C9*1/*5 (P<0.001, Jonckheere-Terpstra test). In summary, the present study demonstrates that, in a Black population, CYP2C9*5, *6, *8 and *11 variants, but not CYP2C9*9, are associated with a decreased phenytoin metabolism. The data also confirm the limited contribution of MDR1 gene to inter-individual phenytoin pharmacokinetic variation. PMID- 16220111 TI - Valproic acid-induced skeletal malformations: associated gene expression cascades. AB - OBJECTIVES: Valproic acid (VPA) is a widely used anticonvulsant medication with well-known teratogenic effects in both humans and in experimental animal model systems. The most commonly observed malformations induced by VPA in experimental animals include neural and skeletal defects. In this study the potential alterations in somitic tissue gene expression relative to the development of observed axial skeletal defects were examined. METHODS: SWV mice were treated at 8.5 days post coitum (d.p.c.) with 1.36 mmol/kg or 2.72 mmol/kg VPA by i.p. injection. At 18.5 d.p.c., animals were killed and stained for morphological and skeletal examination. Cervical malformations consisting of vertebral fusions and cervical ribs were consistently observed. Phenotypic analysis confirmed the presence of dose-dependent axial skeletal malformations induced by in-utero VPA exposure. Using antisense RNA amplification and cDNA microarrays, we examined the expression of approximately 5700 genes in the first six postotic somites of control and treated embryos at 6, 12, 18 and 24 h after the 8.5 d.p.c. VPA treatment. RESULTS: Analysis indicated that several ontological groups (e.g. histone deacetylase complex, guanosine triphosphatases, cell proliferation and cytoskeletal) have significantly enriched gene expression changes in response to the teratogenic insult. The RNA from 6 h post-treatment was also subjected to a microarray cross-platform validation, and genes identified on both platforms are presented. CONCLUSION: These data were then used to deduce candidate cellular pathways that may be responsible for the VPA-induced teratogenic skeletal phenotypes. PMID- 16220112 TI - Thiopurine S-methyltransferase pharmacogenetics: variant allele functional and comparative genomics. AB - Thiopurine S-methyltransferase (TPMT) catalyses the S-methylation of thiopurine drugs. Genetic polymorphisms for TPMT are a major factor responsible for large individual variations in thiopurine toxicity and therapeutic effect. The present study investigated the functional effects of human TPMT variant alleles that alter the encoded amino acid sequence of the enzyme, TPMT*2, *3A, *3B, *3C and *5 to *13. After expression in COS-1 cells and correction for transfection efficiency, allozymes encoded by these alleles displayed levels of activity that varied from virtually undetectable (*3A,*3B and *5) to 98% (*7) of that observed for the wild-type allele. Although some allozymes had significant elevations in apparent Km values for 6-mercaptopurine and S-adenosyl-L-methionine (i.e. the two cosubstrates for the reaction), the level of enzyme protein was the major factor responsible for variation in activity. Quantitative Western blot analysis demonstrated that the level of enzyme protein correlated closely with level of activity for all allozymes except TPMT*5. Furthermore, protein levels correlated with rates of TPMT degradation. TPMT amino acid sequences were then determined for 16 non-human mammalian species and those sequences (plus seven reported previously, including two nonmammalian vertebrate species) were used to determine amino acid sequence conservation. Most human TPMT variant allozymes had alterations of residues that were highly conserved during vertebrate evolution. Finally, a human TPMT homology structural model was created on the basis of a Pseudomonas structure (the only TPMT structure solved to this time), and the model was used to infer the functional consequences of variant allozyme amino acid sequence alterations. These studies indicate that a common mechanism responsible for alterations in the activity of variant TPMT allozymes involves alteration in the level of enzyme protein due, at least in part, to accelerated degradation. PMID- 16220113 TI - St. John's wort and its constituent hyperforin concordantly regulate expression of genes encoding enzymes involved in basic cellular pathways. AB - OBJECTIVES AND METHODS: The effects of St. John's wort and hyperforin on gene expression were analysed in HepG2 cells by Affymetrix microarray hybridization and real time reverse transcription-PCR. RESULTS: Both compounds increased mRNAs of the drug metabolizing enzymes CYP3A4, CYP1A1, CYP1A2 and the flavin containing monooxygenase FMO5, and of the multidrug resistance protein MRP2. CYP4F2 and the reduced nicotinamide adenine dinucleotide dehydrogenase NQO1 were downregulated. Expression of genes mediating cholesterol biosynthesis was decreased, while facilitated glucose transporters and glycolysis genes were induced, indicating increased glucose metabolism. Changes of a considerable number of additional transcripts corresponded to reports on gene regulation by hypoxia. Endoplasmic reticulum stress-regulated genes involved in unfolded protein response and in protection of cells from apoptosis were downregulated. Other calcium binding proteins were affected by both treatments, suggesting an increase in intracellular calcium. CONCLUSIONS: St. John's wort and hyperforin concordantly affected expression of genes not only mediating metabolism and transport of exogenous and endogenous compounds, but also involved in energy metabolism, intracellular calcium regulation, cell proliferation and apoptosis. PMID- 16220114 TI - [Exertional dyspnoea with a normal chest]. PMID- 16220115 TI - [Severe asthma, IgE and anti-IgE: from sceptical waiting to factual learning]. PMID- 16220116 TI - Libyostrongylus infections in ostriches--a brief review with particular reference to their detection in New Zealand. AB - Following the recent discovery of Libyostrongylus douglassii in ostriches in New Zealand, information relating to this nematode is reviewed. Amongst some of the topics considered are the parasite's distribution and prevalence, its life cycle and survival, and its diagnosis and control. Of particular interest from a New Zealand perspective is the potential for the parasite to infect other ratites, especially the kiwi. While the current evidence suggests that this may be unlikely to occur, the possibility that it might be able to do so cannot be ruled out entirely. PMID- 16220117 TI - The welfare significance of the castration of cattle: a review. AB - Castration is an ancient husbandry procedure used to produce docile cattle for draught work, to reduce unwanted breeding, and to modify carcass quality. All the physical methods used to castrate cattle have side-effects and cause pain. The plasma cortisol response to castration using Burdizzo clamps and, by inference, the acute pain experienced, is less than that caused by surgical, rubber-ring or latex-band castration. The cortisol response may be influenced by the age of the animal castrated, but this has not been well defined. Local anaesthesia virtually eliminates the cortisol response, and thus the acute pain, caused by rubber-ring or latex-band castration, but needs to be combined with a systemic analgesic such as the non-steroidal anti-inflammatory drug ketoprofen to eliminate the cortisol response to Burdizzo or surgical castration. When used alone, ketoprofen sometimes reduces the cortisol response to Burdizzo or surgical castration but may need to be accompanied by local anaesthesia to eliminate the pain-induced behaviour seen during the castration process itself. Thus, pharmacological methods are available to virtually eliminate the acute pain experienced by calves during the 12 h following castration. The use of these methods is an additional cost for farmers and may be limited by the availability of drugs for farmers to use and the scarcity of veterinarians in farm animal practice. PMID- 16220118 TI - Pregnancy loss in dairy cattle in the Waikato region of New Zealand. AB - AIM: To define the incidence rate of pregnancy loss and risk factors for those losses in pasture-fed dairy cattle in the Waikato region of New Zealand. METHODS: Cows (n=2,004) from 10 pasture-fed, spring-calving dairy herds in the Waikato were enrolled following confirmation of pregnancy 29-45 days after insemination, for inseminations that occurred within the first 16 days of the seasonal breeding period. Transrectal ultrasonographic examinations for pregnancy were conducted at approximately 6, 8, 10, 14 and 22 weeks gestation, and subsequent calving data were recovered. Pregnancy loss was defined as having occurred when a confirmed pregnancy was not re-diagnosed, when gross abortion was detected, or when a cow calved <265 days after the confirmed conception date. Data were analysed using reverse stepwise logistic regression and Cox's proportional hazards analysis. RESULTS: A total of 128 (6.4%) pregnancy losses were detected. The incidence rate was higher in early compared to late gestation (10.9 vs 2.8 losses/10,000 cow days between Weeks 6-10 vs Weeks 10-14, respectively; p<0.001). Higher rates of loss were associated with the occurrence of clinical mastitis (Hazards ratio (HR)=1.57; p=0.071), being treated for anoestrus (HR=1.69; p=0.007), and in cows that had calving-to-conception intervals < or =63 days compared with those that had calving-to-conception intervals >92 days (HR=2.49; p=0.06). In addition, the rate of pregnancy loss differed between herds (p=0.05). CONCLUSIONS: The highest rate of pregnancy loss occurred in early gestation. Clinical mastitis, anoestrus and calving late in the calving season were risk factors for pregnancy loss. CLINICAL RELEVANCE: Pregnancy diagnosis using ultrasonography can be undertaken from 28 days post-insemination. However, due to the high rate of pregnancy loss at this stage of gestation, herdowners need to be warned of possible losses, and cows should be re-examined to confirm pregnancy before certification of pregnancy status is given. PMID- 16220119 TI - Clearance of para-aminohippuric acid in wethers consuming locoweed. AB - AIM: To validate the use of para-aminohippuric acid (PAH) as a marker for measuring blood flow in wethers consuming a mixed diet of locoweed and blue grama hay. METHODS: Fourteen sheep, stratified by bodyweight (BW), were assigned to one of three treatments: 0.8 mg swainsonine (SW)/kg BW (HI), 0.2 mg SW/kg BW (LO), and no SW (Control). Sheep were fed various ratios of locoweed and blue grama hay to deliver SW treatments, for 28 days prior to infusion of PAH. Concentrations of SW and activities of alkaline phosphatase (Alk-P) and aspartate aminotransferase (AST) in serum were measured to confirm exposure to SW and subclinical intoxication. A single 20-ml injection of 5% PAH was delivered into the jugular vein after subclinical intoxication had been achieved. Blood samples were collected and serum analysed for PAH immediately prior to injection, then every 5 min from 5-30 min, and every 10 min from 30-60 min, following injection of PAH. RESULTS: Effective delivery of SW was evident from the greater concentrations of SW measured in the serum of HI compared with LO animals (p<0.05). No significant differences were detected in the rate of elimination (range 0.097-0.108 L/min), elimination half-life (range 6.62-7.24 min), apparent volume of distribution for the central compartment (range 7.14-9.72 L), and clearance (range 0.73-0.92 L/min) of PAH, between treatments. CONCLUSIONS: Subclinical intoxication with SW did not affect the pharmacokinetics of PAH. Thus, use of downstream dilution of PAH is a valid method to determine the rate of blood flow in nutrient flux experiments that involve consumption of locoweed. PMID- 16220120 TI - Ranging behaviour and duration of survival of wild brushtail possums (Trichosurus vulpecula) infected with Mycobacterium bovis. AB - AIM: To quantify the duration of survival of possums (Trichosurus vulpecula) infected with Mycobacterium bovis, and identify aspects of their behaviour which may influence the likelihood of disease transmission to domestic stock or wildlife. METHODS: Capture and den locations of 14 naturally infected tuberculous possums, eight possums experimentally infected with M. bovis and eight non infected possums were recorded between May 1998 and February 2000 at a study site near Castlepoint on the Wairarapa coast of the North Island in New Zealand. Denning behaviour was observed weekly using radiotelemetry, and possums were captured, examined and released bi-monthly. Data were used to estimate survival period; create denning, activity, and total ranges; and to identify extended forays by possums as individuals and groups. RESULTS: Seventeen tuberculous possum carcasses were recovered, of which 14 (82%) were close to or within their activity range. Denning ranges were known for 10/17 possums that died. Four tuberculous possums were found dead within their denning range. Three possums made extended forays in the 3 weeks before death. Twelve possums were found dead in dense scrub, three in long grass in open woodland and two on pasture. Mean duration of survival of naturally infected possums following detection of clinical signs was 3.4 months (95% CI=2.1-5.4) and the instantaneous mortality rate was 0.293 per month (95% CI=0.184-0.470). Signs of disease were obvious for about 3 weeks prior to death. Tuberculous possums were commonly trapped on only part of the area where the total non-infected population was trapped. CONCLUSION: Most tuberculous possums died within their activity range and in scrub, representing a risk of transmission of M. bovis to wildlife and livestock that forage in scrub. Smaller proportions dying on pasture represent a less frequent, but highly visible risk. Tuberculous possums were clustered on the study site, and localised possum control operations would be more effective if focussed on such areas. PMID- 16220121 TI - Oral vaccination of brushtail possums (Tichosurus vulpecula) with BCG: immune responses, persistence of BCG in lymphoid organs and excretion in faeces. AB - AIMS: To determine immune responses, and the localisation and persistence of Mycobacterium bovis bacille Calmette-Guerin (BCG) in gut-associated lymphoid tissues (GALT) and other organs in possums vaccinated orally with lipid formulated BCG vaccine. To determine the duration of excretion and longevity of survival of BCG in the faeces of vaccinated animals. METHODS: Possums (n=28) were vaccinated with lipid-formulated BCG (1 x 10(8) colony forming units (cfu) of formulated BCG) by the oral route. Control possums (n=17) were fed oral bait pellets containing formulation medium only. Possums were sacrificed at 3 days and at 1, 3, 6 and 8 weeks after vaccination or ingestion of bait. Proliferation responses to bovine purified protein derivative (PPD) were measured in lymphocytes from blood and mesenteric lymph nodes (MLN) and samples of lung, spleen, liver, MLN and Peyer's patches (PP) were cultured for the presence of BCG. The number of BCG organisms excreted in faeces and the duration of excretion were determined in eight vaccinated possums and eight control possums over a 3 week period. In a separate experiment, a further six possums were vaccinated with oral BCG vaccine (5-10 x 10(8) cfu BCG/possum) and their faeces collected over 48 72 h, for culture of BCG. The longevity of survival of BCG in these faeces was determined by storing faecal samples (n=12) under three different conditions: in an incubator (22.5 degrees C), and conditions which simulated the forest floor and open pasture. A proportion (1-2 g) of these faecal samples was collected after storage for 1, 3, 5, 8 or 20 weeks, and cultured for BCG. RESULTS: Possums vaccinated orally with BCG vaccine showed strong proliferation responses to bovine PPD in peripheral blood lymphocytes at 6-8 weeks post-vaccination (p.v.). Positive lymphocyte proliferation assay (LPA) responses to bovine PPD were first evident in MLN at 3 weeks p.v. BCG was cultured from MLN and PP in a proportion of animals at 3-8 weeks p.v. BCG was not cultured from sections of spleen, lung or liver at any time p.v. BCG was recovered in low to moderate numbers from the faeces of vaccinated possums for up to 7 days, and maximal numbers were cultured in faeces collected 48-72 h p.v. After storage for 1 week, BCG was cultured from all faecal samples placed in the incubator and from a proportion of faeces exposed to conditions similar to those on the forest floor and pasture. With the exception of one faecal sample stored under forest floor conditions which was culture-positive for BCG at 3 and 5 weeks, BCG was not cultured from any other faecal sample stored for more than 1 week. CONCLUSIONS: Ingestion of oral BCG vaccine by possums was associated with the development of strong cell-mediated immunity in both blood and MLN. Following oral vaccination with BCG, the organisms were localised and persisted in GALT but did not spread to the spleen, liver or lungs. BCG was shed in low to moderate numbers in the faeces for up to 7 days p.v. The viability of BCG excreted in faeces decreased rapidly, particularly when faeces were exposed to an open pasture environment. Oral vaccination of possums with formulated BCG is unlikely to result in undue contamination of the environment with BCG. PMID- 16220122 TI - Congenital polycystic kidney disease in lambs. AB - AIM: To describe the pathology and inheritance of a congenital polycystic kidney disease (PKD) of sheep. METHODS: Mode of inheritance of PKD was investigated by evaluation of results of the disorder from planned matings in two consecutive years within subsets of a flock that had a high prevalence of PKD in lambs. Gross pathological and histopathological studies were based on tissues derived from this study. Haematoxylin and eosin (H&E)-stained paraffin sections of kidney, liver, extrahepatic biliary and pancreatic ducts, pancreas and epididymis were used to describe the lesions. RESULTS: Twenty-five lambs affected by PKD, of both sexes, were born, numbers in accord with those expected for an autosomal recessive disorder in the population studied. In all cases for which tissues were available, the renal, bile ductal (intrahepatic and extrahepatic), pancreatic and epididymal tissues had widespread dysplastic changes and associated cyst formation. CONCLUSIONS: The findings of renal cysts in conjunction with cysts in other organs are unifying features in many of the human and animal forms of PKD and suggest a related pathogenic and genetic base consistent with an autosomal recessive disorder. PMID- 16220123 TI - The occurrence of Cryptosporidium parvum, Campylobacter and Salmonella in newborn dairy calves in the Manawatu region of New Zealand. AB - AIM: To determine the occurrence of Cryptosporidium parvum oocysts, Campylobacter spp and Salmonella spp in faecal samples taken from newborn dairy calves on 24 dairy farms in the Manawatu region of New Zealand. METHODS: A cross-sectional study was conducted during the 2002 calving season. Faecal samples were collected from 185 newborn calves from a convenience sample of 24 dairy farms. The samples were tested microscopically for the presence of C. parvum oocysts, and bacteriologically for the presence of Campylobacter spp and Salmonella spp. RESULTS: Infections with C. parvum were identified in 33/156 (21.2%) calves from 10 farms. More than 10(6) oocysts/g (OPG) faeces were detected in calves from four farms. Campylobacter spp were isolated from 58/161 (36%) calves from 18 farms; in particular, C. jejuni subsp jejuni was isolated from 11/161 (6.8%) calves from seven farms. Salmonellae were not detected. CONCLUSIONS: Despite the short and concentrated calving pattern and the long interval between calving seasons characterising most dairy farms in New Zealand, C. parvum is widespread among calves. Campylobacter spp, especially C. jejuni, rapidly colonise the intestinal tract of newborn calves. RELEVANCE: This study provided an estimate of the ecological impact of newborn dairy calves with regard to the potentially zoonotic enteric pathogens most frequently isolated from human gastrointestinal infections in New Zealand. PMID- 16220124 TI - The effect of progesterone supplementation post mating and shearing of ewes in early pregnancy on the reproductive performance of ewes and birthweight of lambs. AB - AIM: To determine whether short-term progesterone supplementation post mating or shearing of ewes in early pregnancy affected either the proportion of ewes that lambed or that had multiple lambs, or the birthweight of lambs. METHODS: Romney ewes (n=457) were synchronised in oestrus using controlled internal drug releasing (CIDR) devices containing progesterone, and mated to Romney rams over a 5-day period. The mid-point of mating (Day 0) occurred 2 days after the withdrawal of CIDR devices. Ewes mated (n=397) were randomly allocated to one of four treatment groups: shearing at Day 5, shearing at Day 30, no shearing, and no shearing plus progesterone supplementation using a CIDR device inserted on Day 3 for 6 days. During the period from Day 5 to Day 27, six harnessed Suffolk rams were placed with the ewes and matings recorded. At Day 48, all ewes that did not return to the Suffolk rams were scanned for pregnancy using ultrasound. At Day 140, single- and multiple-bearing ewes were set-stocked at 15.1 and 12.2 ewes/ha, respectively, and equivalent numbers of ewes from each treatment group were placed in each paddock. Blood samples from 10 unshorn and 10 progesterone supplemented ewes were collected on Days 3, 6 and 9, and analysed for plasma progesterone concentrations. Lambs were identified to their dam and weighed within 12 h of birth, and again at 27 and 93 days after the mid-point of lambing. The ewes were weighed at regular intervals throughout the trial. RESULTS: Plasma progesterone concentrations of supplemented ewes were higher than those of unsupplemented ewes (3.28 vs 1.75 ng/ml) on Day 6 (p=0.02) but not on Day 9 (4.58 vs 4.63 ng/ml). Treatment of ewes had no effect on either the proportion of ewes which lambed to the synchronised mating period or that had multiple lambs. Lambs born to ewes shorn at Day 30 tended (p=0.09) to be heavier at birth (by 0.28 kg) than those born to unshorn ewes but this effect was not evident when data were corrected for length of gestation. Neither shearing at Day 5 nor progesterone supplementation had any effect on the birthweight of lambs, and the treatment of ewes had no effect on the survival rate of lambs to weaning. CONCLUSIONS: Progesterone supplementation for 6 days beginning 3 days post mating did not increase either the proportion of ewes that lambed or that had multiple lambs, or the birthweight of lambs. Shearing 5 days after mating had no significant effect on the reproductive performance of ewes and need not be avoided, but is unlikely to result in an increase in lamb birthweight. Shearing ewes at Day 30 may result in an increase in the birthweight of lambs but, ideally, ewes should be further advanced in pregnancy before shearing is undertaken. PMID- 16220126 TI - Prediction of penicillin resistance in Staphylococcus aureus isolates from dairy cows with mastitis, based on prior test results. AB - AIM: To gauge how well prior laboratory test results predict in vitro penicillin resistance of Staphylococcus aureus isolates from dairy cows with mastitis. METHODS: Population-based data on the farm of origin (n=79), genotype based on pulsed-field gel electrophoresis (PFGE) results, and the penicillin-resistance status of Staph. aureus isolates (n=115) from milk samples collected from dairy cows with mastitis submitted to two diagnostic laboratories over a 6-month period were used. Data were mined stochastically using the all-possible-pairs method, binomial modelling and bootstrap simulation, to test whether prior test results enhance the accuracy of prediction of penicillin resistance on farms. RESULTS: Of all Staph. aureus isolates tested, 38% were penicillin resistant. A significant aggregation of penicillin-resistance status was evident within farms. The probability of random pairs of isolates from the same farm having the same penicillin-resistance status was 76%, compared with 53% for random pairings of samples across all farms. Thus, the resistance status of randomly selected isolates was 1.43 times more likely to correctly predict the status of other isolates from the same farm than the random population pairwise concordance probability (p=0.011). This effect was likely due to the clonal relationship of isolates within farms, as the predictive fraction attributable to prior test results was close to nil when the effect of within-farm clonal infections was withdrawn from the model. CONCLUSIONS: Knowledge of the penicillin-resistance status of a prior Staph. aureus isolate significantly enhanced the predictive capability of other isolates from the same farm. In the time and space frame of this study, clinicians using previous information from a farm would have more accurately predicted the penicillin-resistance status of an isolate than they would by chance alone on farms infected with clonal Staph. aureus isolates, but not on farms infected with highly genetically heterogeneous bacterial strains. PMID- 16220125 TI - Bodyweights and growth rates of spring- and autumn-born Thoroughbred horses raised on pasture. AB - AIM: To examine the growth of spring- and autumn-born Thoroughbred foals raised on pasture. METHODS: Bodyweight and growth rates were measured in pasture-raised Thoroughbred horses, born in either spring (n=56) or autumn (n=7), from birth to approximately 13 and 17 months of age. RESULTS: Birthweight tended to be lower in autumn- than spring-born foals (54.4, SD 7.92 kg vs 57.3, SD 5.90 kg; p=0.08). Between birth and 6 months of age, there was no difference in growth rate at equivalent ages between horses born in spring and autumn. Spring-born horses, which were weaned in the autumn, had lower post-weaning growth rates than autumn born horses that were weaned in the spring. At time of the late yearling sales (March-April) in the Southern Hemisphere, unadjusted mean bodyweights of autumn born horses (379.3, SD 24.8 kg) were lower (p=0.017) than those of the spring born horses (437.2, SD 35.3 kg), although values in the autumn-born horses were all within two standard deviations (SD) of the mean of the spring-born animals. When adjusted for the covariates of birthweight and gender, the difference between spring- and autumn-born horses at that time was not significant (p=0.25). CONCLUSIONS AND CLINICAL RELEVANCE: Some autumn-born foals could be marketed for late yearling sales in the Southern Hemisphere, on the basis of bodyweight. Furthermore, they might also be competitive in the Northern Hemisphere industry (sales or racing), as they would be competing against horses of the same official age. PMID- 16220127 TI - Does sward height grazed by ewes in mid- to late-pregnancy affect indices of colostrum intake by twin and triplet lambs? AB - AIMS: To determine if sward height grazed by ewes in mid- to late-pregnancy affects subsequent indices of colostrum intake by twin and triplet lambs, and the extent of the variation in colostrums intake between twin and triplet litter mates. METHODS: Blood samples were collected from 75 twin and 46 triplet litters, respectively, 24-36 h after birth. Samples were not collected from lambs that died before this age. Lambs were born to ewes which had grazed one of four sward heights (2, 4, 6 and 8 cm) during mid- to late-pregnancy. Serum samples were analysed for gamma-glutamyl transferase (GGT) activities and glucose concentrations. The number of lambs present at weaning was used to determine lamb survival rate. RESULTS: GGT activities of lambs born to ewes that grazed swards 2 cm high were lower than those born to ewes that grazed swards 4 and 6 cm high (p=0.053 and p=0.037, respectively). Glucose concentrations were higher in twin than triplet lambs (p=0.003) and in lambs born to ewes grazed on swards 6 cm high than those born to ewes grazed on swards 2 cm high (p=0.033). These differences were not evident after correction for differences in birthweight. There was significant (p<0.001) variation within litter-mates for GGT activities and glucose concentrations in both twins and triplets. The blood constituent status of lambs that died before the 24-36 h collection period is not known. However, lambs which died after this period had significantly lower GGT activities (p=0.009) and glucose (p=0.010) concentrations at the time of sampling than those which survived. CONCLUSIONS: Farmers should ensure twin- and triplet-bearing ewes in mid- to late-pregnancy are grazing at least 4 cm of herbage to ensure colostrum intake of lambs in the postpartum period is not adversely affected. PMID- 16220128 TI - The effect of individual liveweight and use of teaser rams prior to mating on the reproductive performance of ewe hoggets. AB - AIM: To determine the effects of liveweight (LW) and use of teaser rams prior to mating on both the mating performance and pregnancy rate of ewe hoggets. METHODS: Romney hoggets (n=841) were weighed and randomly assigned to one of two treatment groups; either exposed to seven harnessed, vasectomised (teaser) rams for 17 days prior to the breeding period (n=283); or not exposed to teaser rams (unteased, n=558). Harnessed Perendale rams were then introduced for two oestrous cycles. Using crayon marks, hoggets were identified as having been mated during the first 17 days only, during the second 17 days only, during both periods, or not mated. All mated hoggets were then scanned for pregnancy using ultrasound, 48 days after the end of the breeding period, and identified as being either non-pregnant, or as single-, twin or triplet-bearing. RESULTS: Hoggets mated in either the first 17 days only or in both the first and second 17-day periods were heavier than those mated in the second period only (p<0.001). Those mated in the first or second 17-day period only, and those mated in both periods were all heavier prior to mating than those not mated at all (p<0.001). Twin-bearing hoggets were heavier than both their single-bearing and non-pregnant counterparts (p<0.001). A greater proportion of the teased hoggets were mated in the first 17 days only compared with unteased hoggets (62.6 vs 32.1%; p<0.001). In contrast, more of the unteased hoggets were marked in the second 17-day period only (p<0.001). A greater proportion of mated, unteased hoggets were found to be non-pregnant in comparison to their teased counterparts (17.8 vs 11.2%; p=0.02). CONCLUSIONS: Heavier hoggets at breeding were more likely to be mated in the first 17 days of the breeding period and more likely to be twin-bearing. Use of teaser rams for 17 days prior to the breeding period increased the proportion of hoggets mated in the first 17 days, increased conception rates, and thus increased the proportion of pregnant hoggets. These results indicate that farmers should utilise teasers for 17 days prior to commencement of the breeding period and avoid mating hoggets that weigh below 36 kg. PMID- 16220129 TI - Confirmation of resistance to ivermectin by Cooperia curticei in sheep. AB - AIM: To investigate the occurrence of resistance to a full dose of oral ivermectin by Cooperia curticei in sheep. METHODS: Twelve lambs on a sheep and cattle property in the North Island of New Zealand were randomly allocated to one of two equal-sized groups. One group was treated orally with a single dose of ivermectin at the manufacturer's recommended dose rate of 0.2 mg/kg, while the other remained as an untreated control. Worm counts were carried out post mortem on the abomasa and small intestines of all animals in both groups 7 days after treatment. RESULTS: While treatment with ivermectin reduced the numbers of all other worm genera to almost zero, those of Ostertagia(= Telodorsagia) circumcincta and C. curticei were reduced by only 37% and 19%, respectively. CONCLUSIONS: These results provide clear evidence of resistance to ivermectin by O. circumcincta and C. curticei. They also appear to represent the first record of macrocylic lactone (ML) resistance in C. curticei in sheep in New Zealand or elsewhere. PMID- 16220130 TI - Spirometra erinacei / S. erinaceieuropaei in a feral cat in Manawatu with chronic intermittent diarrhoea. AB - CASE HISTORY: A feral cat captured in the Manawatu region of New Zealand was treated for worms and fleas, and kept confined in a metabolic cage. It showed good appetite and weight gain but had intermittent watery, yellow diarrhoea. CLINICAL FINDINGS: Clinical examination under sedation was unremarkable and routine blood tests showed no significant abnormalities. The cat was negative for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Different canned cat foods did not alter the course of the diarrhoea, and the cat was euthanised 6 months after capture. At necropsy, two sections of adult Spirometra tapeworms were found in the jejunum and typical Spirometra eggs were found in colonic contents. Molecular identification of the parasite was undertaken, using the cytochrome-c oxidase subunit-1 gene (cox1) sequence. DIAGNOSIS: Chronic intermittent diarrhoea associated with Spirometra erinacei / S. erinaceieuropaei infection. CLINICAL RELEVANCE: Spirometra has not been reported in New Zealand before but has been associated with gastrointestinal disease in cats in other parts of the world. It requires species targeted treatment to be eliminated effectively, and is zoonotic. Diagnosis could be difficult for clinicians who are not familiar with the parasite and its life cycle. PMID- 16220131 TI - Suspected Vestia foetida poisoning in young goats. AB - CASE HISTORY: Two crossbred, castrated male goats, a 5-month-old and an 8-month old, were observed ingesting Vestia foetida (Solanaceae). Later, the goats were seen standing splay-legged and apparently disoriented. CLINICAL FINDINGS: When examined, both goats were in sternal recumbency and had mydriasis; the younger goat had a diminished menace response. When the goats were made to stand, they were ataxic and had muscle fasciculations of the hindquarters and face. Both had halitosis consistent with the odour of crushed Vestia leaves. The animals were treated with a mixture of vitamins and intravenous diazepam. The older goat recovered but the younger goat died and was necropsied. This animal had severe periacinar necrosis and fatty change in the liver, as well as fatty nephrosis. DIAGNOSIS: Probable Vestia foetida poisoning. CLINICAL RELEVANCE: The introduction of Vestia foetida to New Zealand and the apparent palatability of the plant necessitate that veterinarians and owners be knowledgeable about its potential toxicity. Differential diagnoses for the liver lesions (in New Zealand) would include Cestrum poisoning, acute seneciosis, acute blue-green algal poisoning, and acute and chronic copper poisoning. PMID- 16220132 TI - Lesions in the brains of three cattle resembling the lesions of enterotoxaemia in lambs. AB - CASE HISTORY: A 3-month-old female Angus calf was found dead, and two adult Friesian dairy cows died soon after developing nervous signs. PATHOLOGICAL FINDINGS: Grossly, bilateral and mostly symmetrical areas of haemorrhage were evident that mainly involved areas of grey matter in the brainstem from the level of the caudal colliculi to the thalamus and, in one, the internal capsule and caudate nucleus. In the occipital and caudal parietal cortex, there was extensive oedema of white matter. Histologically, in addition to haemorrhage, there was protein-rich oedema around arterioles and venules in the cerebrum, hippocampus, internal capsule, thalamus, midbrain, dorsal medulla, and central cerebellar and cerebellar folial white matter. The calf 's brain had bilateral and symmetrical oedema and necrosis affecting several brainstem nuclei and the occipital grey matter overlying areas of oedema of the corona radiata. DIAGNOSIS: Although the cause was not established, the perivascular lesions resembled those produced in calves by the intravenous administration of epsilon toxin. CLINICAL RELEVANCE: It is possible that epsilon toxin-induced enterotoxaemia occurs naturally in cattle, and where bilateral haemorrhage is recognised in the brains of cattle, small intestinal contents should be collected for analysis of epsilon toxin. PMID- 16220134 TI - Mycotic dermatitis with digital gangrene and osteomyelitis, and protozoal intestinal parasitism in Marlborough green geckos (Naultinus manukanus). AB - CASE HISTORY: Thirty adult Marlborough green geckos (Naultinus manukanus) were collected from Stephens Island and held over winter, prior to their translocation. Five adult geckos developed skin lesions after husbandry changes affected the humidity of their enclosures. Two geckos underwent ecdysis and recovered. One animal died and two others progressively worsened and were presented for treatment. CLINICAL AND PATHOLOGICAL FINDINGS: The geckos were in poor body condition and had multiple black powdery lesions and solitary raised white nodules on their skin. Both geckos died despite topical and supportive treatment. Histopathology showed the skin nodules contained branching non-septate hyphae infiltrating necrotic epidermal tissue, and associated dermal inflammation. There was necrosis of several digits and mycotic osteomyelitis. Mucor ramosissimus was cultured from skin biopsies from each animal. Large numbers of motile protozoa, resembling Trichomonas, and another unidentifiable, were recovered from fresh faecal smears, and Nyctotherus sp protozoa were present in the lumen of the intestine of one animal post mortem. DIAGNOSIS: Mycotic dermatitis with digital gangrene and osteomyelitis due to Mucor ramosissimus, and enteric protozoal parasitism with Trichomonas sp and Nyctotherus sp. CLINICAL RELEVANCE: The clinical course and pathological findings of mycotic dermatitis in two Marlborough green geckos involved in a wildlife translocation in New Zealand are reported, and also the first record of the Marlborough green gecko as a host for the enteric protozoa Trichomonas sp and Nyctotherus sp. PMID- 16220133 TI - Suspected fatal venous air embolism during anaesthesia in a Pomeranian dog with pulmonary calcification. AB - CASE HISTORY: Death occurred in a 1.25 kg, 9-month-old female Pomeranian dog undergoing anaesthesia for surgical repair of partially healed fractures of the radius and ulna. CLINICAL FINDINGS: Following sedation, anaesthesia was induced using thiopentone and maintained with halothane in oxygen. An acute decrease in the dog's end-tidal carbon dioxide (EtCO2) measurements was noted approximately 50 min after induction, immediately following delivery of a 5-ml bolus of lactated Ringer's solution (LRS) administered to flush a small (0.06 ml) volume of fentanyl via a pre-placed intravenous (I/V) catheter. Venous air embolism (VAE) was suspected and the dog died despite interventive therapy. On post-mortem examination, several air bubbles were noted when the right ventricle was opened under water. Histologically, the kidneys appeared abnormal with immature glomeruli, and the lungs appeared diffusely mineralised. The origin of the air was probably small bubbles and microbubbles that may have been present in the extension set and 20 ml syringe used for the administration of fentanyl and I/V fluids to the dog. DIAGNOSIS: Renal dysplasia and diffuse pulmonary calcification, with VAE as the probable cause of death. CLINICAL RELEVANCE: In this case of VAE-associated anaesthetic death, it is further speculated that underlying pulmonary disease, in the form of pulmonary calcification, may have contributed to an increased sensitivity to the adverse effects of VAE. PMID- 16220135 TI - Cerebral infarction and meningoencephalitis following hot-iron disbudding of goat kids. AB - CASE HISTORY: Twelve of 150 goat kids, 4-10 days old, died 3 days after disbudding with a hot iron. Another 18 kids had been ill the previous day but survived following antibiotic therapy. Five of the dead kids were necropsied. PATHOLOGICAL FINDINGS: There was necrosis and haemorrhage of the skin, subcutaneous tissues and frontal bone at disbudding sites in all five kids examined post mortem. Beneath disbudding sites in 4/5 kids there were bilateral, dark red, often cavitated areas of necrosis extending deep into the frontal cortex of the brain. Histologically, these areas consisted of coagulation necrosis, haemorrhage, vascular thrombosis and suppurative inflammation. Numerous bacteria, predominantly large Gram positive rods, were present in the necrotic brain tissue. In the remaining kid, bilateral areas of yellow discolouration and flattening of gyri in frontal lobes corresponded histologically to extensive polioencephalomalacia. A mixed growth of aerobes and anaerobes was cultured from the brain of one kid with suppurative lesions. CLINICAL RELEVANCE: Thermal disbudding of neonatal kids is widely practised in dairy goat herds and is considered the method of choice for disbudding in New Zealand. However, the skull of goat kids is much thinner than that of calves and the safety margin for thermal injury to the brain is markedly reduced. This report highlights the risks associated with the technique and its potential as a welfare issue. PMID- 16220137 TI - Study of intramammary infections in dairy goats from mountainous regions in Italy. PMID- 16220136 TI - Concurrent gastro-oesophageal intussusception, trichobezoar and hiatal hernia in a cat. AB - CASE HISTORY: An adult male Birman cat was evaluated for recurrent, intermittent vomiting or regurgitation, occasionally associated with abdominal discomfort. CLINICAL FINDINGS AND DIAGNOSIS: Radiographs, including an oesophogram, indicated an oesophageal obstruction. Prior to treatment, the cat's condition deteriorated and it was euthanised at the owner's request. Post-mortem examination revealed a gastro-oesophageal intussusception, a trichobezoar impacted into the intussusceptum, and a dilated oesophageal hiatus consistent with a chronic hiatal hernia. CLINICAL RELEVANCE: Gastro-oesophageal intussusception is a rare condition in cats. Its aetiology in relation to a pre-existing hiatal hernia and a trichobezoar is discussed. PMID- 16220140 TI - Points of view: content versus process: is this a fair choice? Undergraduate biology courses for nonscientists: toward a lived curriculum. PMID- 16220141 TI - Applying instructional design theories to bioinformatics education in microarray analysis and primer design workshops. AB - The need to support bioinformatics training has been widely recognized by scientists, industry, and government institutions. However, the discussion of instructional methods for teaching bioinformatics is only beginning. Here we report on a systematic attempt to design two bioinformatics workshops for graduate biology students on the basis of Gagne's Conditions of Learning instructional design theory. This theory, although first published in the early 1970s, is still fundamental in instructional design and instructional technology. First, top-level as well as prerequisite learning objectives for a microarray analysis workshop and a primer design workshop were defined. Then a hierarchy of objectives for each workshop was created. Hands-on tutorials were designed to meet these objectives. Finally, events of learning proposed by Gagne's theory were incorporated into the hands-on tutorials. The resultant manuals were tested on a small number of trainees, revised, and applied in 1-day bioinformatics workshops. Based on this experience and on observations made during the workshops, we conclude that Gagne's Conditions of Learning instructional design theory provides a useful framework for developing bioinformatics training, but may not be optimal as a method for teaching it. PMID- 16220142 TI - An inquiry into protein structure and genetic disease: introducing undergraduates to bioinformatics in a large introductory course. AB - This inquiry-based lab is designed around genetic diseases with a focus on protein structure and function. To allow students to work on their own investigatory projects, 10 projects on 10 different proteins were developed. Students are grouped in sections of 20 and work in pairs on each of the projects. To begin their investigation, students are given a cDNA sequence that translates into a human protein with a single mutation. Each case results in a genetic disease that has been studied and recorded in the Online Mendelian Inheritance in Man (OMIM) database. Students use bioinformatics tools to investigate their proteins and form a hypothesis for the effect of the mutation on protein function. They are also asked to predict the impact of the mutation on human physiology and present their findings in the form of an oral report. Over five laboratory sessions, students use tools on the National Center for Biotechnology Information (NCBI) Web site (BLAST, LocusLink, OMIM, GenBank, and PubMed) as well as ExPasy, Protein Data Bank, ClustalW, the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and the structure-viewing program DeepView. Assessment results showed that students gained an understanding of the Web-based databases and tools and enjoyed the investigatory nature of the lab. PMID- 16220143 TI - Students investigating the antiproliferative effects of synthesized drugs on mouse mammary tumor cells. AB - The potential for personalized cancer management has long intrigued experienced researchers as well as the naive student intern. Personalized cancer treatments based on a tumor's genetic profile are now feasible and can reveal both the cells' susceptibility and resistance to chemotherapeutic agents. In a weeklong laboratory investigation that mirrors current cancer research, undergraduate and advanced high school students determine the efficacy of common pharmacological agents through in vitro testing. Using mouse mammary tumor cell cultures treated with "unknown" drugs historically recommended for breast cancer treatment, students are introduced to common molecular biology techniques from in vitro cell culture to fluorescence microscopy. Student understanding is assessed through laboratory reports and the successful identification of the unknown drug. The sequence of doing the experiment, applying logic, and constructing a hypothesis gives the students time to discover the rationale behind the cellular drug resistance assay. The breast cancer experiment has been field tested during the past 5 yr with more than 200 precollege/undergraduate interns through the Gains in the Education of Mathematics and Science program hosted by the Walter Reed Army Institute of Research. PMID- 16220144 TI - How effective are simulated molecular-level experiments for teaching diffusion and osmosis? AB - Diffusion and osmosis are central concepts in biology, both at the cellular and organ levels. They are presented several times throughout most introductory biology textbooks (e.g., Freeman, 2002), yet both processes are often difficult for students to understand (Odom, 1995; Zuckerman, 1994; Sanger et al., 2001; and results herein). Students have deep-rooted misconceptions about how diffusion and osmosis work, especially at the molecular level. We hypothesized that this might be in part due to the inability to see and explore these processes at the molecular level. In order to investigate this, we developed new software, OsmoBeaker, which allows students to perform inquiry-based experiments at the molecular level. Here we show that these simulated laboratories do indeed teach diffusion and osmosis and help overcome some, but not all, student misconceptions. PMID- 16220145 TI - Using a module-based laboratory to incorporate inquiry into a large cell biology course. AB - Because cell biology has rapidly increased in breadth and depth, instructors are challenged not only to provide undergraduate science students with a strong, up to-date foundation of knowledge, but also to engage them in the scientific process. To these ends, revision of the Cell Biology Lab course at the University of Wisconsin-La Crosse was undertaken to allow student involvement in experimental design, emphasize data collection and analysis, make connections to the "big picture," and increase student interest in the field. Multiweek laboratory modules were developed as a method to establish an inquiry-based learning environment. Each module utilizes relevant techniques to investigate one or more questions within the context of a fictional story, and there is a progression during the semester from more instructor-guided to more open-ended student investigation. An assessment tool was developed to evaluate student attitudes regarding their lab experience. Analysis of five semesters of data strongly supports the module format as a successful model for inquiry education by increasing student interest and improving attitude toward learning. In addition, student performance on inquiry-based assignments improved over the course of each semester, suggesting an improvement in inquiry-related skills. PMID- 16220147 TI - [Post-traumatic stress disorder and substance use disorder: epidemiology, nature and neurobiology]. AB - Post-traumatic stress disorder (PTSD) is frequently associated with other psychiatric diagnoses, being substance use disorder (SUD) one of the most prevailing. Comorbid disorders in general and SUD in particular frequently complicate the course and outcome of PTSD, and vice versa. As with other dual disorders, comorbidity of PTSD and SUD is complex, having been proposed different theoretical models to explain it that will be discussed in the present paper, the first of a series of two. In addition, the present paper reviews data from epidemiological studies, as well as the clinical characteristics, the relevance of stressful life events as a vulnerability factor to PTSD as well as to SUD and the neurobiological basis of the association of both disorders with the aim of gaining a better understanding of this comorbidity, so that the treatment of this dual disorder will be discussed in the second part. PMID- 16220146 TI - Increased fidelity reduces poliovirus fitness and virulence under selective pressure in mice. AB - RNA viruses have high error rates, and the resulting quasispecies may aid survival of the virus population in the presence of selective pressure. Therefore, it has been theorized that RNA viruses require high error rates for survival, and that a virus with high fidelity would be less able to cope in complex environments. We previously isolated and characterized poliovirus with a mutation in the viral polymerase, 3D-G64S, which confers resistance to mutagenic nucleotide analogs via increased fidelity. The 3D-G64S virus was less pathogenic than wild-type virus in poliovirus-receptor transgenic mice, even though only slight growth defects were observed in tissue culture. To determine whether the high-fidelity phenotype of the 3D-G64S virus could decrease its fitness under a defined selective pressure, we compared growth of the 3D-G64S virus and 3D wild type virus in the context of a revertible attenuating point mutation, 2C-F28S. Even with a 10-fold input advantage, the 3D-G64S virus was unable to compete with 3D wild-type virus in the context of the revertible attenuating mutation; however, in the context of a non-revertible version of the 2C-F28S attenuating mutation, 3D-G64S virus matched the replication of 3D wild-type virus. Therefore, the 3D-G64S high-fidelity phenotype reduced viral fitness under a defined selective pressure, making it likely that the reduced spread in murine tissue could be caused by the increased fidelity of the viral polymerase. PMID- 16220148 TI - [Functional status and quality of life in Latin American outpatients with schizophrenia treated with atypical or typical antipsychotics: outcomes of the 12 months schizophrenia outpatient health outcomes (IC-SOHO) Study]. AB - OBJECTIVE: Functional status and quality of life outcomes in Latin American outpatients with schizophrenia were compared after 12 months of monotherapy treatment with olanzapine, risperidone or typical antipsychotics. METHOD: Both outcomes were assessed as part of a prospective, large (N= 7658), international (27 countries), observational study. RESULTS: from the Latin American subpopulation (N= 2671; 11 countries) are presented. Compared to typical antipsychotics, olanzapine and risperidone were associated with significantly (p < 0.05) greater odds of employment and social activity, and significantly greater improvements in quality of life. Olanzapine was also associated with significantly greater odds of living independently, compared to typical antipsychotics. CONCLUSION: This study indicates that functional status and quality of life outcomes are likely to be more favorable when Latin American outpatients with schizophrenia are treated with olanzapine or risperidone monotherapy, rather than typical antipsychotics. PMID- 16220149 TI - [The mind -- body problem]. AB - In this paper I present, in the first place, two of the main problems in the philosophy of mind: 1) the problem of the criteria of the mental, and 2) the problem of the relationship between the mental and the physical. In the last part, I mention two problems highly discussed nowadays which, in my view, will produce the most important advance in the knowledge of our minds in the next decades: the problem of consciousness and its neural basis, and the problem of the origin and development of a theory of mind. PMID- 16220150 TI - [Neurosciences and philosophy of mind]. AB - In this paper we argue that the interaction between neurosciences and philosophy of the mind is on the way to understand consciousness, and to solve the mind-body or mind-brain problem. Naturalism is the view that mental processes are just brain processes and that consciousness is a natural phenomenon. It is possible to construct a theory about its nature by blending insights from neuroscience, philosophy of the mind, phenomenology, psychology and evolutionary biology. PMID- 16220151 TI - [Models of self knowledge]. AB - The main purpose of this paper is to analyze some different explanatory models of self - knowledge, belonging to Contemporary analytic philosophy. As a starting point, I will focus on self-ascriptions of mental states of the likes of "I have a headache", "I am thinking about my son", "I desire to shock my father", namely, the so-called "avowals". In the first part, I will point out what I take to be the set of characteristics of avowals that any theory about self-knowledge should account for. In the second part, I will present and contrast with one another the main theoretical explanatory models that have been put forward to give the required account. PMID- 16220152 TI - [Wittgenstein and Freud]. AB - In this paper I present the most important limits in the reading that Ludwig Wittgenstein made of Sigmund Freud. In particular, starting from certain thematic axes (distinction causes / reasons, conception of the meaning, interpretation concept, the problem of the truth in the therapeutic treatment, the paper of the patient's conviction), I try to show that the position of Wittgenstein in front of the Freudian psychoanalysis was ambiguous: on one hand he conceives it as an example more than "philosophical mythology", but on the other hand, he is identified with the psychoanalyst and he is conceived itself as making something related with that an analyst makes. PMID- 16220153 TI - [The primordial genesis of psyche: emergence and formalization]. AB - Although there is no doubt that life was preceded by a purely physical universe from which it emerged, still there are some conceptual blanks which are necessary to analyse in order to make intelligible not only the evolutional origin of sensibility, perception and conscience but also its settlement in the process of evolution and the contribution of psychicsm to the efficiency of organisms as tele-onomical systems. Nowadays science considers that this happens because of a continuous emergence of novelty in organisms with a more and more developed and complex nervous system, which facilitates actions of adaptation seeking optimal survival. According to Zubiri, the basque thinker who we intend to analyse here, the humanization springs up from potentiality of matter, through a process of transformation, systematization and elevation. Intelligence emerges as a result of biological hyper-formalization so that man can survive taking over reality. If this function is increased, it will give richness to psychic life and, as this is a changeable activity, it can admit enlargements, retractions and internal modifications. PMID- 16220154 TI - [Body and soul: fragments]. PMID- 16220155 TI - [From the seduction theory to the oedipus complex]. AB - The author reviews the Freudian theory of seduction as it was presented in the last decade of the XIX century. Freud began to talk about the effects of the seduction in the clinical history of Katherine, (Studies on Hysteria, 1893 - 1895). In 1896 in Heredity and the Aetiology of the Neuroses, Freud considered the seduction as the specific cause of the psychoneurosis, and latter in The Aetiology of Hysteria, he separated himself from Breuer and Charcot. In Further Remarks on The Neuro - Psychoses of Defense, Freud relates seduction with repression. The author thinks that his father death makes him doubt this theory. These oscillations went hand in hand with his auto-analysis, as it is related in the letters of October 3 and 15, 1897. In these letters the love to the mother and the jealousy to the father are introduced as key features. Nevertheless, the theory of seduction comes back recurrently in the works of Freud, and as late as 1906, in My Views on the Part Played by Sexuality in the Aetiology of the Neurose, he seems to down play the importance as a aetiology feature, considering that the hysterical patient falsified his memories and replaced them with fantasies. Towards the end of his life, in An Outline of Psycho - Analysis, Freud stated that the cares of the mother makes her in the first seducer of the child. According to Etchegoyen (2003), Freud never abandoned the theory of seduction. PMID- 16220156 TI - Cultivating crystal forms. PMID- 16220157 TI - Some recent developments in the chemical synthesis of inorganic nanotubes. AB - Inorganic nanotubes have been a subject of intensive research in the past decade. We recently developed a number of synthetic strategies for generating nanotubes from inorganic materials that do not have a layered structure. It is the intention of this contribution to provide a brief account of these research activities. PMID- 16220158 TI - An organic-inorganic hybrid material constructed from a three-dimensional coordination complex cationic framework and entrapped hexadecavanadate clusters. AB - A unique organic-inorganic hybrid compound has been separated under hydrothermal condition, which is constructed from a three-dimensional second metal-organic subunit and entrapped hexadecavanadate clusters. PMID- 16220159 TI - Connecting cyclophosphazenes via ring N-centres with covalent linkers. AB - Two cyclotriphosphazene rings can be covalently linked via ring N-centres by a 2 butene-1,4-diyl unit and vice versa a cyclotriphosphazene molecule is able to bridge two cinnamyl groups via two ring N-centres yielding in either case dicationic assemblies which offers a route to novel polycations. PMID- 16220160 TI - Slow relaxation of magnetisation in an octanuclear cobalt(II) phosphonate cage complex. AB - The synthesis, structural and magnetic characterisation of a new polymetallic cobalt complex are reported; the magnetic behaviour is unusual. PMID- 16220161 TI - New examples of metalloaromatic Al-clusters: (Al4M4)Fe(CO)3 (M = Li, Na and K) and (Al4M4)2Ni: rationalization for possible synthesis. AB - Ab initio calculations reveal that all-metal antiaromatic molecules like Al4M4 (M = Li, Na and K) can be stabilized in half sandwich (Al4M4)Fe(CO)3 and full sandwich (Al4M4)2Ni complexes. The formation of the full sandwich complex [(Al4M4)2Ni] from its organometallic precursor depends on the stability of the organic-inorganic hybrid (C4H4)Ni(Al4Li4). PMID- 16220162 TI - Coexistence of ferro- and antiferromagnetic interactions in a metal-organic radical-based (6,3)-helical network with large channels. AB - A metal-organic open-framework with an unprecedented (6,3)-helical topology, large channels and mixed ferro- and antiferromagnetic interactions has been synthesized using a three-connecting tricarboxylic polychlorotriphenylmethyl radical and Co(ii) ions. PMID- 16220163 TI - Synthesis, structure and magnetic properties of a decametallic Ni single-molecule magnet. AB - Ferromagnetic exchange between the ten Ni2+ ions in the complex [Ni10(tmp)2(N3)8(acac)6(MeOH)6] leads to a spin ground state of S = 10; single crystal M vs. H studies reveal the temperature and sweep rate dependent hysteresis loops expected for a single-molecule magnet. PMID- 16220164 TI - The folded, tetrameric phosph(III)azane macrocycle [(P(mu-NtBu))2(1,4 (NH)2C6H4)]4. AB - The tetrameric macrocycle [(P(mu-NtBu))2(1,4-(NH)2C6H4)]4, obtained from the reaction of the phosphazane dimer [ClP(mu-NtBu)]2 with p-phenylenediamine, has an unusual folded conformation in the solid state and contains a roughly tetrahedral arrangement of endo N-H groups for the potential coordination of anions. PMID- 16220165 TI - Stereoisomerism in polyoxometalates: structural and spectroscopic studies of bis(malate)-functionalized cluster systems. AB - The tetrameric macrocycle [(P(mu-NtBu))2(1,4-(NH)2C6H4)]4, obtained from the reaction of the phosphazane dimer [ClP(mu-NtBu)]2 with p-phenylenediamine, has an unusual folded conformation in the solid state and contains a roughly tetrahedral arrangement of endo N-H groups for the potential coordination of anions. PMID- 16220166 TI - Triplet ground state (S= 1) pegylated bis(aminoxyl) diradical: synthesis and the effect of water on magnetic properties. AB - The synthesis and magnetic characterization of pegylated bis(aminoxyl) diradical with an S= 1 ground state are presented, revealing water-induced changes in the molecular conformation and magnetic properties. PMID- 16220167 TI - Borromean sheets assembled by self-supporting argentophilic interactions. AB - An infinite two-dimensional Borromean coordination framework, stabilized by argentophilic interactions, was obtained by the reaction of a flexible ligand with AgBF4. PMID- 16220169 TI - Synthesis of bis(bora)calix[4]arenes bearing perfluoroaryl substituents. AB - New routes to perfluoroaryl complexes of bis(bora)calix[4]arenes are described; crystallographic and spectroscopic data are presented. PMID- 16220168 TI - Homologous, long-chain alkyl dendrons form homologous thin films on silver oxide surfaces. AB - As suggested by X-ray crystal structures, homologous, long-chain alkyl dendrons with three carboxyl groups form thin films on silver oxide surfaces, which give reflection-absorption infrared spectra that show a linear increase in intensities of methylene C-H stretching absorptions. PMID- 16220171 TI - Immobilization of enzymes on a microchannel surface through cross-linking polymerization. AB - A novel and facile method for the preparation of an enzyme-immobilized microreactor has been developed in which enzymes are immobilized as an enzyme polymer membrane formed on the inner wall of the microchannel by a cross-linking polymerization method; the resulting microreactor shows excellent reaction performance and stability against denaturating agents. PMID- 16220170 TI - Silver nanoparticles fabricated in Hepes buffer exhibit cytoprotective activities toward HIV-1 infected cells. AB - Silver nanoparticles fabricated in Hepes buffer exhibit potent cytoprotective and post-infected anti-HIV-1 activities toward Hut/CCR5 cells. PMID- 16220172 TI - Regioselective aromatic C-H silylation of five-membered heteroarenes with fluorodisilanes catalyzed by iridium(I) complexes. AB - The aromatic C-H silylation of five-membered heteroarenes with 1,2-di-tert-butyl 1,1,2,2-tetrafluorodisilane regioselectively proceeded at 120 degrees C in octane in the presence of a catalytic amount of iridium(I) complexes generated from 1/2[Ir(OMe)(COD)]2 and 2-tert-butyl-1,10-phenanthroline. PMID- 16220173 TI - Multiple high-level QM/MM reaction paths demonstrate transition-state stabilization in chorismate mutase: correlation of barrier height with transition state stabilization. AB - Multiple profiles for the reaction from chorismate to prephenate in the enzyme chorismate mutase calculated with hybrid density functional combined quantum mechanics/molecular mechanics methods (B3LYP/6-31G(d)-CHARMM27) agree well with experiment, and provide direct evidence of transition-state stabilization by this important enzyme, which is at the centre of current debates about the nature of enzyme catalysis. PMID- 16220174 TI - Binding interactions between the host cucurbit[7]uril and dendrimer guests containing a single ferrocenyl residue. AB - The stability of the inclusion complexes formed between the host cucurbit[7]uril and dendrimers containing a single ferrocene residue is strongly affected by the solution pH and the growth of the dendrimer, reaching its highest values on the second and third generation dendrimers, whereas no complex is formed with the first generation compound. PMID- 16220176 TI - Negative magnetoresistance in Ba2CoS3. AB - A small negative magnetoresistance and metallic-like behaviour has been detected for the first time in a one-dimensional sulfide containing Co2+. PMID- 16220175 TI - Identification of new N-Sb topologies: understanding the sequential dehydrochloride coupling of primary amines and trichloropnictines. AB - Subtle steric strain imposed by 2,6-dimethylphenyl substituents on N-Sb frameworks has enabled identification of the first acyclic dipnictadiazane and the first six-membered cyclotristibatriazane providing insight into the dehydrohalide coupling reaction of amines with halopnictines. PMID- 16220177 TI - Trialkylsulfonium dicyanamides--a new family of ionic liquids with very low viscosities. AB - Trialkylsulfonium dicyanamides show surprisingly low viscosities down to -20 degrees C and are therefore highly interesting liquid materials for separation processes and electrolyte applications at low temperatures. PMID- 16220178 TI - Domino Michael addition-carbene rearrangement-cyclization reaction of 1 alkynyl(aryl)-lambda3-bromanes with 2-mercapto-1,3-benzazoles. AB - Exposure of 1-alkynyl[p-(trifluoromethyl)phenyl](tetrafluoroborato)-lambda3 bromanes to 2-mercaptobenzimidazole or benzothiazole in dichloromethane at 0 degrees C under argon resulted in a domino Michael addition-carbene rearrangement cyclization reaction to produce directly tricyclic heterocycles in high yields, whereas the reaction with 2-mercaptobenzoxazole afforded 1-alkynyl sulfides. PMID- 16220179 TI - One-step labelling of sphingolipids via a scrambling cross-metathesis reaction. AB - The alkyl chain in the backbone of sphingosine derivatives can be exchanged with functionalised (labelled) side chains in a single step under cross-metathesis reaction conditions. PMID- 16220180 TI - Iterative, orthogonal strategy for oligosaccharide synthesis based on the regioselective glycosylation of polyol acceptors with partially unprotected n pentenyl-orthoesters: further evidence for reciprocal donor acceptor selectivity (RDAS). AB - An efficient iterative, orthogonal protocol based on the regioselective glycosyl coupling of D-mannose polyols with, partially unprotected, n-pentenyl orthoester donors permits the synthesis of linear and branched oligosaccharides with minimal protecting group tampering. PMID- 16220181 TI - Synthesis and characterization of side-bound aryldiazo and end-bound nitrosyl complexes of nickel. AB - Structural characterization of a nickel aryldiazo complex supported by the bulky 1,2-bis(di-tert-butylphosphino)ethane ligand reveals square-planar Ni geometry and an unusual side-on coordination mode for the N2R moiety, while the related nitrosyl complex displays trigonal-planar geometry at Ni and end-on coordination of the NO ligand. PMID- 16220182 TI - Carbon nanotube-promoted Co-Cu catalyst for highly efficient synthesis of higher alcohols from syngas. AB - The development of a type of carbon-nanotube-promoted Co-Cu catalyst, which displays excellent performance for highly effective and selective formation of the C(2-4)-oxygenates, especially BuOH and DME, from syngas, is reported. PMID- 16220183 TI - Total synthesis of a diastereomer of the marine natural product clavosolide A. AB - The first total synthesis of the reported structure of the sponge metabolite clavosolide A is described using a Prins cyclisation to assemble the tetrahydropyran core followed by manipulation of the side-chain, dimerisation and finally glycosidation. PMID- 16220184 TI - Synthesis of 2-alkylidenetetrahydrofurans by Ru-catalyzed regio- and stereoselective codimerization of dihydrofurans with alpha,beta-unsaturated esters. AB - 2-(1-Alkoxycarbonyl)alkylidenetetrahydrofurans were readily synthesized by the codimerization of 2,3- or 2,5-dihydrofurans with alpha,beta-unsaturated esters using a zerovalent Ru catalyst, Ru(cod)(cot), with high regio- and stereoselectivity. PMID- 16220185 TI - Copper(II)-catalysed addition of O-H bonds to norbornene. AB - Cu(OTf)2 is an inexpensive, air- and moisture-stable catalyst for the O-H addition of aliphatic and aromatic acids and alcohols to norbornene. PMID- 16220186 TI - A formal synthesis of (+)-lactacystin. AB - A synthetic route to the neurotrophic agent (+)-lactacystin has been developed utilizing a base-promoted intramolecular alkylidenecarbene C-H insertion as the key transformation. PMID- 16220187 TI - Stannyl cyclopropanes by diastereoselective cyclopropanations with (tributylstannyl)-diazoacetate esters catalyzed by Cu(I) N-heterocyclic carbene. AB - Catalyzed cyclopropanations of alkenes with Bu3SnC(=N2)CO2R (R = Et, t-Bu) have been achieved in good yield with excellent diastereoselectivity to make stannyl cyclopropanes having two or three stereocenters, one of which is quaternary. PMID- 16220188 TI - Crystalline Na-Si(NN) derivatives [Si(NN)= Si((NCH2tBu)2C6H4-1,2)]: the silylenoid [Si(NN)OMe]-, the dianion [(NN)Si-Si(NN)]2-, and the radical anion c [Si(NN)]3-. AB - Reactions of the silylene Si[(NCH2Bu(t))2C6H4-1,2], [Si(NN)], with NaOMe, excess Na or 1/3 Na yield the X-ray-characterised crystalline compounds [Na(micro Si(NN)OMe)(THF)(OEt2)]2 (2b), [Na(THF)2Si(NN)]2 (3) and [Na(THF)4][(Si(NN))3-c] (4). PMID- 16220189 TI - Exploiting C3-symmetry in the dynamic coordination of a chiral trisoxazoline to copper(II): improved enantioselectivity, and catalyst stability in asymmetric lewis acid catalysis. AB - Chiral C3-symmetric trisoxazolines are highly efficient stereodirecting ligands in enantioselective Cu(II) Lewis acid catalysis which is based on the concept of a stereoelectronic hemilability of the divalent copper; in direct comparison with the analogous bisoxazoline systems they are more efficient in the enantioselective alpha-amination as well as the enantioselective Mannich reaction of prochiral beta-ketoesters. PMID- 16220190 TI - [Blockade of NMDA receptor enhances corticosterone-induced downregulation of brain-derived neurotrophic factor gene expression in the rat hippocampus through cAMP response element binding protein pathway]. AB - High concentration of corticosterone leads to morphological and functional impairments in hippocampus, ranging from a reversible atrophy of pyramidal CA3 apical dendrites to the impairment of long-term potentiation (LTP) and hippocampus-dependent learning and memory. Glutamate and N-methyl-D-aspartate (NMDA) receptor play an important role in this effect. Because of the importance of brain-derived neurotrophic factor (BDNF) in the functions of the hippocampal neurons, alteration of the expression of BDNF is thought to be involved in the corticosterone effect on the hippocampus. To determine whether change in BDNF in the hippocampus is involved in the corticosterone effect, we injected corticosterone (2 mg/kg, s.c.) to Sprague-Dawley rats and measured the mRNA, proBDNF and mature BDNF protein levels in the hippocampus. We also measured the phosphorylation level of the transcription factor cAMP response element binding protein (CREB). Furthermore, we intraperitoneally injected NMDA receptor antagonist MK801 (0.1 mg/kg) 30 min before corticosterone administration to investigate whether and how MK801 affected the regulation of BDNF gene expression by corticosterone. Our results showed that 3 h after single s.c. injection of corticsterone, the expression of BDNF mRNA, proBDNF and mature BDNF protein decreased significantly (P<0.01). MK801 promoted the downregulation of BDNF gene expression in the rat hippocampus by corticosterone. We also found that either applying corticosterone or co-applying corticosterone with MK801 downregulated the phosphoration level of CREB, the latter (corticosterone plus MK801) being more effective (P<0.05). Taken together, our results indicate that corticosterone downregulates BDNF gene expression in the rat hippocampus through CREB pathway and that blockade of NMDA receptor enhances this effect of corticosterone in reducing BDNF expression. PMID- 16220191 TI - Activation of p38 mitogen-activated protein kinase in spinal cord contributes to chronic constriction injury-induced neuropathic pain. AB - The present study aimed to investigate the role of spinal p38 mitogen-activated protein kinase (p38 MAPK) activation in chronic constriction injury (CCI) of the sciatic nerve induced neuropathic pain. CCI model was produced by loosely ligating the left sciatic nerve proximal to the sciatica's trifurcation with 4-0 silk thread in male Sprague-Dawley rat. SB203580, a specific inhibitor of the p38 MAPK, was intrathecally administered on day 5 post-CCI. Thermal and mechanical nociceptive thresholds were assessed with the paw withdrawal lantency (PWL) to radiant heat and the paw withdrawal threshold (PWT) to von Frey filaments respectively. The protein levels of the phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated cAMP response element binding protein (pCREB) were assessed by Western blot analysis. The results showed that CCI significantly increased the expressions of cytosolic and nuclear p-p38 MAPK in the spinal cord. Intrathecal administration of SB203580 dose-dependently reversed the established mechanical allodynia and thermal hyperalgesia induced by CCI. Correlated with behavior results, SB203580 dose-dependently inhibited the CCI-induced increase of the expressions of cytosolic and nuclear p-p38 MAPK and nuclear pCREB in the spinal cord. Taken together, these findings suggest that the activation of p38 MAPK pathway contributes to the development of neuropathic pain induced by CCI, and that the function of p-p38 MAPK may partly be accomplished via the CREB-dependent gene expression. PMID- 16220192 TI - An investigation on the division of neuronal PC12 cells induced by nerve growth factor. AB - Neuronal PC12 cells induced by nerve growth factor (NGF) have been considered to be postmitotic and lack the ability to divide. However, in this study, we not only detected DNA synthesis but also observed cell division in some morphologically differentiated neuronal PC12 cells bearing long neurites. More interestingly, in addition to the division of perikaryon, the neurites located on the division site of the cell membrane also divided into two parts and were allocated to the two daughter cells. These results demonstrate that the morphologically differentiated neuronal PC12 cells still retain the ability to divide. This is the first report that neuronal PC12 cells as well as their neurites can divide. PMID- 16220193 TI - [Activation of the spinal extracellular signal-regulated kinase is involved in morphine dependence and naloxone-precipitated withdrawal response]. AB - Extracellular signal-regulated kinase (ERK), a mitogen-activated protein kinase (MAPK), transduces a broad range of extracellular stimuli into diverse intracellular responses. It has been reported that ERK is involved in the modulation of nociceptive information and central sensitization produced by intense noxious stimuli or peripheral tissue inflammation. Our previous studies showed that the spinal neurons sensitization was involved in morphine withdrawal response. This study was to investigate the role of the spinal ERK in morphine dependence and naloxone-precipitated withdrawal response. To set up morphine dependent model, rats were subcutaneously injected with morphine (twice a day, for 5 d). The dose of morphine was 10 mg/kg on the first day and was increased by 10 mg/kg each day. On day 6, 4 h after the injection of morphine (50 mg/kg), morphine withdrawal syndrome was precipitated by an injection of naloxone (4 mg/kg, i.p.). Using anti-phospho-ERK (pERK) antibody, the time course of pERK expression was detected by Western blot. U0126, a mitogen-activated protein kinase kinase (MEK) inhibitor, or phosphorothioate-modified antisense oligonucleotides (ODN) was intrathecally injected 30 min or 36, 24 and 12 h before naloxone-precipitated withdrawal. The scores of morphine withdrawal symptom and morphine withdrawal-induced allodynia were observed. One hour after naloxone-precipitated withdrawal, pERK expression in the spinal dorsal horn was assessed by immunohistochemical analysis and Western blot was used to detect the expression of cytosolic and nuclear fraction of pERK in the rat spinal cord. The results showed that the expression of cytosolic and nuclear fraction of pERK, not non-phospho-ERK, in the spinal cord was gradually increased following the injection of morphine. When morphine withdrawal was precipitated with naloxone, the expression of the spinal pERK further increased. Intrathecal administration of U0126 or antisense ODN against ERK decreased the scores of morphine withdrawal, attenuated morphine withdrawal-induced allodynia and also inhibited the increase of pERK expression in the spinal cord of morphine withdrawal rats. These results suggest that activation of the spinal ERK is involved in morphine dependent and naloxone-precipitated withdrawal response. PMID- 16220194 TI - Migration of intravenously grafted mesenchymal stem cells to injured heart in rats. AB - The present study aimed to determine the role of tissue injury in migration of mesenchymal stem cells (MSCs) intravenously transplanted into heart and to establish experimental basis for improving stem cell therapy in its targeting and effectiveness. MSCs were isolated from bone marrow of male Sprague-Dawley rats and purified by density centrifuge and adhered to the culture plate in vitro. Female rats were divided randomly into four groups. Myocardial ischemia (MI) transplanted group received MSCs infusion through tail vein 3 h after MI and compared with sham-operated group or normal group with MSCs infusion, or control group received culture medium infusion. MI was created in female rats by ligating the left anterior descending coronary artery. The heart was harvested 1 week and 8 weeks after transplantation. The characteristics of migration of MSCs to heart were detected with expression of sry gene of Y chromosome by using fluorescence in situ hybridization (FISH). Ultrastructural changes of the ischemic myocardium of the recipient rats were observed by transmission electron microscope (TEM). One week or 8 weeks after transplantation, sry positive cells were observed in the cardiac tissue in both of MI transplanted group and sham-operated group, the number of sry positive cells being significantly higher in MI transplanted group (P<0.01). No significant difference was found in the number of sry positive cells between 1 week and 8 weeks after transplantation. No sry positive cells were observed in the hearts of control and normal group. In addition, the ultrastructure of some cells located in the peri-infarct area of MI rats with MSCs transplantation was similar to that of MSCs cultured in vitro. These results indicate that MSCs are capable of migrating towards ischemic myocardium in vivo and the fastigium of migration might appear around 1 week after MI. The tissue injury and its degree play an important role in the migration of MSCs. PMID- 16220195 TI - Characteristic neuronal firing interspike intervals in laterodorsal thalamic nuclei induced by tetanization of rat caudate putamen: possible relations to hippocampal electroencephalogram changes. AB - The purpose of the present work was to study the effect of acute tetanization of the right caudate putamen nucleus (ATRC) on single neuronal interspike intervals (ISIs) in both laterodorsal thalamic nuclei (LDi), and electroencephalogram (EEG) wave interpeak intervals (IPIs) in both hippocampi (HPCi). Experiments were performed on 21 male Sprague-Dawley rats weighing 150~250 g. The seizures were induced by the ATRC (60 Hz, 2 s, 0.4~0.6 mA). Quadruple recordings were simultaneously carried out: two for single unit recordings from both LDi, and two for EEG recordings from both HPCi. The ATRC induced: (1) An interactive epileptic electrical network reconstructed in bilateral HPCi, which was driven by primary afterdischarges of single LD neuron. (2) A symmetric mirror-like ISI spot distribution of the LD neuronal firing before and after tetanus. (3) Gradually prolonged LD neuronal discharge intermittence was coherent with synchronous hippocampal EEG activities on the contralateral side. (4) Single LD neuronal spikes were phase- and time-locked to 20~25 Hz gamma oscillations in contralateral HPC. It suggests a particular temporal code patterning of single LD neuronal firing and its relationships to hippocampal EEG wave code in time series, the latter implies the LD neuronal encoding mechanisms of ATRC-induced epileptic electrical network in bilateral HPCi. PMID- 16220196 TI - [Effects of angiotensin II on extracellular signal-regulated protein kinases signaling pathway in cultured vascular smooth muscle cells from Wistar-Kyoto rats and spontaneously hypertensive rats]. AB - The aim of this study was to investigate the effects of angiotensin II (Ang II) on extracellular signal-regulated protein kinase (ERK) signaling pathway in cultured vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. VSMCs from SHR and WKY rats were treated with 1x10(-7) mmol/L Ang II for 24 h in the absence or presence of 30 min of pre treatment of valsartan (1x10(-5) mmol/L) or PD98059 (1x10(-5)mmol/L), selective inhibitor of ERKs- dependent pathways, when they were cultured in 20% calf serum medium. VSMCs of SHR and WKY cultured in serum-free medium were used as control groups. Among the different treatments, VSMCs from the SHR and WKY were devided into four groups: (1) control, (2) Ang II, (3) Ang II + valsartan, (4) Ang II + PD98059. ERK activity in VSMCs was measured by immuno-precipitation. Proteins of total ERK (t-ERK), phosphorylated-ERK (p-ERK) and mitogen-activated protein kinases phosphatase-1 (MKP-1) in VSMCs were detected by Western blot. MKP-1 mRNA in VSMCs was measured by RT-PCR. In VSMCs from WKY or SHR rats, ERK activity, p ERK, MKP-1 and MKP-1 mRNA in Ang II group were higher than those in control group (P<0.05). In both SHRs and WKYs, there were no significant differences in ERK activity, p-ERK, MKP-1 and MKP-1 mRNA among the control group, Ang II + valsartan group and Ang II + PD98059 group. ERK activity, p-ERK, MKP-1 and MKP-1 mRNA in SHRs were significantly higher than those in WKYs with same treatments (P<0.01). There was no significant difference in t-ERK among different groups and no difference in t-ERK between SHRs and WKYs (P>0.05). Our results show that Ang II activates VSMCs ERK signaling pathways via Ang II type 1 (AT(1)) receptors. Ang II increased ERK activity and p-ERK, but not t-ERK, accompanied by an increase in MKP-1 mRNA expression and protein. Among the different treatments, ERK activity and p-ERK were higher in SHR than in WKY. Valsartan and PD98059 blocked Ang II stimulated ERK activation. These results suggest that ERK signaling pathway plays an important role in the pathogenesis of hypertension. The effect of Ang II on SHR and WKY VSMCs' ERK signaling pathway may be mediated by AT(1) receptors, enhancing ERK activity and the amount of p-ERK, and then increasing MKP-1 mRNA and its expression. PMID- 16220197 TI - [Electrophysiological effects of neurotransmitters on pacemaker cells in guinea pig left ventricular outflow tract]. AB - This study was designed to explore the innervation of autonomic nervous system and the distribution of receptors on pacemaker cell membrane in guinea pig left ventricular outflow tract (aortic vestibule). By using conventional intracellular microelectrode technique to record action potentials, autonomic neurotransmitters and antagonists were used to investigate the electrophysiological features and regularities of spontaneous activity of left ventricular outflow tract cells. Electrophysiological parameters examined were: maximal diastolic potential (MDP), amplitude of action potential (APA), maximal rate of depolarization (V(max)), velocity of diastolic depolarization (VDD), rate of pacemaker firing (RPF), 50% and 90% of duration of action potential (APD(50) and APD(90)). The results are listed below: (1) Perfusion with 100 mumol/L isoprenaline (Iso) resulted in a significant increase in V(max) (P <0.05), VDD, RPF, and APA (P <0.01), a notable decrease in MDP (P<0.05), and also a marked shortening in APD(50) (P<0.01). Pretreatment with Iso (100 mumol/L), propranolol (5 mumol/L) significantly decreased RPF and VDD (P<0.01), decreased APA, MDP and V(max) (P<0.01) notably, prolonged APD(50) (P<0.01) and APD(90) (P<0.05) markedly. (2) Application of 100 mumol/L epinephrine (E) resulted in a significant increase in VDD (P<0.05), RPF (P<0.001), V(max) (P<0.05) and APA (P<0.001), and a notable shortening in APD(50) and APD(90) (P<0.05). (3) Perfusion with 100 mumol/L norepinephrine (NE) led to a significant increase in VDD, RPF, APA and V(max) (P<0.05), and a marked shortening in APD(50) (P<0.05). Pretreatment with NE (100 mumol/L), phentolamine (100 mumol/L) significantly decreased RPF and VDD, MDP and APA (P<0.01), decreased V(max) notably (P<0.05), prolonged APD(50) and APD(90) markedly (P<0.01). (4) During perfusion with 10 mmol/L acetylcholine (ACh), VDD and RPF slowed down notably (P<0.05), APA decreased significantly (P<0.001), V(max) slowed down notably (P<0.01), APD50 shortened markedly (P<0.05), Atropine (10 mmol/L) antagonized the effects of ACh (10 mumol/L) on APD(50) (P<0.05). These results suggest that there are probably alpha-adrenergic receptor (alpha-AR), beta-adrenergic receptor (beta-AR) and muscarinic receptor (MR) on pacemaker cell membrane of left ventricular outflow tract in guinea pig. The spontaneous activities of left ventricular outflow tract cells are likely regulated by sympathetic and parasympathetic nerves. PMID- 16220198 TI - Resveratrol reduces intracellular free calcium concentration in rat ventricular myocytes. AB - Resveratrol (trans-3, 4', 5-trihydroxy stilbene), a phytoalexin found in grape skins and red wine, has been reported to have a wide range of biological and pharmacological properties. It has been speculated that resveratrol may have cardioprotective activity. The objective of our study was to investigate the effects of resveratrol on intracellular calcium concentration ([Ca(2+)](i)) in rat ventricular myocytes. [Ca(2+)](i) was detected by laser scanning confocal microscopy. The results showed that resveratrol (15~60 mumol/L) reduced [Ca(2+)](i) in normal and Ca(2+)-free Tyrode's solution in a concentration dependent manner. The effects of resveratrol on [Ca(2+)](i) in normal Tyrode's solution was partially inhibited by pretreatment with sodium orthovanadate (Na3VO4, 1.0 mmol/L, P<0.01), an inhibitor of protein tyrosine phosphatase, or L type Ca(2+) channel agonist Bay K8644 (10 mumol/L, P<0.05), but could not be antagonized by NO synthase inhibitor L-NAME (1.0 mmol/L). Resveratrol also markedly inhibited the ryanodine-induced [Ca(2+)](i) increase in Ca(2+)-free Tyrode's solution (P<0.01). When Ca(2+) waves were produced by increasing extracellular Ca(2+) concentration from 1 to 10 mmol/L, resveratrol (60 mumol/L) could reduce the velocity and duration of propagating waves, and block the propagating waves of elevated [Ca(2+)](i). These results suggest that resveratrol may reduce the [Ca(2+)](i) in isolated rat ventricular myocytes. The inhibition of voltage-dependent Ca(2+) channel and tyrosine kinase, and alleviation of Ca(2+) release from sarcoplasmic reticulum (SR) are possibly involved in the effects of resveratrol on rat ventricular myocytes. These findings could help explain the protective activity of resveratrol against cardiovascular disease. PMID- 16220199 TI - ERK1/2 signaling pathway is involved in 15-hydroxyeicosatetraenoic acid-induced hypoxic pulmonary vasoconstriction. AB - Hypoxia-induced 15-hydroxyeicosatetraenoic acid (15-HETE) is an essential mediator to constrict pulmonary arteries (PA). The signaling pathway involved in 15-HETE-induced PA vasoconstriction remains obscure. The aim of the present study was to test the hypothesis that hypoxic PA constriction induced by 15-HETE was possibly regulated by the extracellular signal-regulated kinase-1/2 (ERK1/2) pathway. PA ring tension measurement, Western blot and immunocytochemistry were used in the study to determine the possible role of ERK1/2 in 15-HETE-induced PA vasoconstriction. The organ bath for PA rings tension study was employed. Adult male Wistar rats were raised in hypoxic environment with fractional inspired oxygen (FIO2, 0.12) for 9 d. PA 1~1.5 mm in diameter were dissected and cut into 3 mm long rings for tension study. ERK1/2 up-stream kinase (MEK) inhibitor PD98059, which blocks the activation of ERK1/2, was used. The results showed that pretreatment of PD98059 significantly blunted 15-HETE-induced PA vasoconstrictions in the rings from hypoxic rat. Moreover, in endothelium-denuded rings, PD98059 also significantly attenuated 15-HETE-induced vasoconstriction. Phosphorylation of ERK1/2 in pulmonary arterial smooth muscle cells (PASMCs) of rat was enhanced evidently when stimulated by 15-HETE. Thus, the data suggest that ERK1/2 signaling pathway is involved in 15-HETE-induced hypoxic pulmonary vasoconstriction. PMID- 16220200 TI - 15-hydroxyeicosatetraenoic acid depressed endothelial nitric oxide synthase activity in pulmonary artery. AB - 15-hydroxyeicosatetraenoic acid (15-HETE) plays an important role in hypoxia induced pulmonary vasoconstriction. Release of nitric oxide (NO) is apparently decreased and activity of endothelial nitric oxide synthase (eNOS) is impaired in chronic hypoxia. However, little is known whether 15-HETE contributes to eNOS/NO pathway in the constriction induced by 15-HETE. We examined the response of rat pulmonary artery (PA) rings to 15-HETE, the production of NO, total eNOS expression and the phosphorylation of eNOS in bovine pulmonary artery endothelial cells (BPAECs) stimulated by 15-HETE. Rat PA rings were divided into three groups: endothelium intact group, endothelium denuded group, and nitro-L-arginine methyl ester (L-NAME, 0.1 mmol/L, an inhibitor of eNOS) group. Constrictions to 15-HETE were significantly enhanced in endothelium denuded group and L-NAME group (both P< 0.05 vs endothelium intact group, n= 9); BPAECs were incubated in different conditions to test nitrite production by Greiss method. Nitrite production was significantly reduced by 1 mumol/L 15-HETE (P<0.05), and increased by the lipoxygenase inhibitors, 10 mumol/L cinnamyl 3,4- dihydroxy-[alpha] cyanocinnamate (CDC, P< 0.05) and 0.1 mmol/L nordihydroguiairetic acid (NDGA, P< 0.01 ); Western blot analysis of extracts from BPAECs incubated with 15-HETE in different time was carried out to test total eNOS expression, and the expression was changed unobviously. Immunoprecipitation (IP) and Western blot analysis of cell extracts from BPAECs treated with 2 mumol/L 15-HETE in different length of time were accomplished, using phospo-eNOS-threonine 495 (Thr495, an inhibitory site) antibody for IP, and eNOS or 15-lipoxygenase (15-LO) antibodies for Western blot. 15-HETE depressed eNOS activity by increasing the levels of phospho-eNOS Thr 495. The data suggest that eNOS/NO pathway is involved in PA constrictions induced by 15-HETE and that 15-HETE depresses eNOS activity by phosphorylation in Thr495 site. The protein interaction between phospho-eNOS (Thr495) and 15-LO is discovered for the first time. PMID- 16220201 TI - Modulatory action of endogenous and exogenous nitric oxide on survival of alveolar macrophages from normal and bleomycin-treated rats. AB - To investigate the modulatory action of endogenous and exogenous nitric oxide (NO) on survival of alveolar macrophages (AMs) in different cellular states, AMs from normal rats (normal AMs) and from bleomycin (BLM)-treated rats (BLM AMs) were incubated by sodium nitroprusside (SNP, NO donor) and L-arginin (L-Arg, NO precursor), respectively. The survival of AMs was evaluated by apoptosis and cell cycles. The molecular mechanisms were investigated by the contents of Bcl-2, Bax proteins in AMs. The results are as follows: (1) The degree of BLM AMs apoptosis was higher than that of normal AMs; the number of BLM AMs in G(0)/G(1) phases was less than that of normal AMs; there was no significant difference in S+G(2)M phases between the number of BLM AMs and that of normal AMs. (2) Down-regulation of Bcl-2 and up-regulation of Bax occurred in BLM AMs, compared to those in normal AMs. (3) Apoptosis of AMs, either normal AMs or BLM AMs, was induced by both SNP and L-Arg, when compared to their respective control; only the number of BLM AMs in S+G(2)M phases was increased by L-Arg. (4) SNP and L-Arg induced a down-regulation of Bcl-2 and an up-regulation of Bax proteins in normal AMs, but did not induce the same change pattern in BLM AMs. (5) The Bax in BLM AMs was down-regulated by L-Arg. It is concluded that NO can induce the apoptosis of BLM AMs and normal AMs; that Bcl-2 and Bax are implicated in NO-induced apoptosis of normal AMs, whereas they are not involved in that of BLM AMs, suggesting the differential molecular mechanisms underlying the NO-induced apoptosis of normal AMs and BLM AMs; and that endogenous NO promotes proliferation of BLM AMs, which might be associated with down-regulation of Bax. PMID- 16220202 TI - [17beta-estradiol protects against injury of aortic relaxation and contraction in ovariectomized rats with insulin resistance induced by fructose]. AB - The purpose of the present study was to investigate the effect of 17beta estradiol (17beta-E(2)) on the structure and relaxation and contraction activity of thoracic aortas in ovariectomized rats with insulin resistance induced by fructose. Ovariectomized mature female Sprague-Dawley rats were fed with high fructose diet for 8 weeks to induce insulin resistance. Physiological dose of 17beta-E(2) (30 mug/kg) was injected subcutaneously every day for 8 weeks. Systolic blood pressure (SBP) was measured by use of tail-cuff. Serum nitric oxide (NO), estradiol (E(2)), fasting blood sugar (FBS) and fasting serum insulin (FSI) were measured respectively in each group. The insulin sensitive index (ISI) was calculated. The thoracic aortas were fixed in formalin, sliced and HE dyed. The structure of thoracic aortas, lumen breadth, media thickness, media thickness/lumen breadth ratio and media cross-section area were measured. The contraction response of thoracic aorta rings induced by L-phenylephrine (PE) and the relaxation response of thoracic aorta rings induced by ACh and sodium nitroprusside (SNP) were measured. To explore the mechanism, nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) was used. The results obtained are as follows: (1) 17beta-E(2) protected against the effect of high fructose diet, which caused an increase in SBP, hyperinsulinemia and a decrease in ISI in ovariectomized rats. (2) The structure of thoracic aortas had no significant difference among the groups. (3) Compared with the ovariectomized group (OVX) or fructose fed group (F), serum nitric oxide was significantly reduced, the contraction response of thoracic aorta rings to PE was enhanced and the relaxation response to ACh was depressed significantly in ovariectomized+fructose fed group (OVX+F). The effect of high fructose was reversed by 17beta-E(2). After pretreatment with L-NAME, the effect of 17beta E(2), which enhanced the relaxation response of thoracic aorta rings to ACh in ovariectomized+fructose+17beta-E(2) group (OVX+F+E(2)), was partly blocked. (4) The relaxation response of thoracic aorta rings to SNP had no significant difference among the groups. (5) The contraction response of thoracic aorta rings without endothelium to PE had no significant difference among the groups. These findings suggest that 17beta-E(2) may provide protection against the effect of high fructose diet, which causes hypertension, dysfunction of endothelial cells and insulin resistance. The mechanism of this effect of 17beta-E(2) could be partly associated with the increase of NO by NOS pathway, or associated with the decrease in the level of systolic blood pressure and serum insulin, and the improvement of insulin resistance. PMID- 16220203 TI - [Relationship between apoptosis and alteration of the energetic metabolism pathways of hypertrophic cardiomyocytes induced by hypoxia-reoxygenation]. AB - The apoptosis of cardiomyocytes plays a pivotal role in the pathogenesis of cardiac failure transformed from cardiac hypertrophy, so that suppression of cardiomyocytes apoptosis is an effective pharmacotherapeutic target to prevent cardiac failure. This study focused on the relationship between apoptosis and alteration of the energetic metabolism pathways of hypertrophic cardiomyocytes induced by hypoxia-reoxygenation. Cardiomyocyte hypertrophy was induced by angiotensin II (0.1 mumol/L ) and norepinephrine (1 mumol/L), and the cells were cultured under the condition of hypoxia ( 95% N2 and 5% CO2, the O2 partial pressure was regulated at least lower than 5 mmHg ) for 8 h, then were recovered to normal culture environment. Apoptosis was detected with TUNEL. The activity of pyruvate dehydrogenase (PDH) and carnitine palmitoyltransferase 1 (CPT-1), the rate of glycose oxidation and glycolysis, and fatty acid metabolism were detected by liquid scintillation counting. The results are as follows: (1) The activity of active PDH (PDHa) was slightly higher in hypertrophic cardiomyocytes than that in normal cardiomyocytes, but the activity of CPT-1 was significantly lower in hypertrophic cardiomyoctes than that in normal cardiomyocytes.Compared with the hypertrophic cardiomyocytes cultured with normal oxygen concentration, the activities of PDHa and CPT-1 were decreased significantly after hypoxia for 8 h, and the activity of PDHa were decreased further after reoxygenation for 4 h, but the activity of CPT-1 recovered quickly after reoxygenation. (2) The rate of glucose oxidation in hypertrophic cardiomyocytes increased slightly when cultured under normal O2 partial pressure than that in normal cardiac cells. The rate of glucose oxidation reduced (16 +/- 0.9)% and (48 +/- 1.1)% in normal and hypertrophic cardiomyocytes, respectively, after hypoxia. It reduced further in hypertrophic cardiac cells at 4 h of reoxygenation, then recovered gradually. In normal cardiocytes, it recovered quickly after reoxygenation. (3) The rate of glycolysis of hypertrophic cardiocytes increased slightly than that of the normal cardiocytes when cultured in the general O(2) environment. Compared with the normal cardiomyocytes, the rate of glycolysis of hypertrophic cardiac cells was the same during hypoxia-reoxygenation culture, i.e., the rate of glycolysis decreased slightly after hypoxia for 8 h, but increased rapidly and significantly after reoxygenation. (4) The rate of fatty acid oxidation was slightly lower in hypertrophic cardiocytes than that in normal cardiomyocytes. After hypoxia for 8 h, the rate of fatty acid oxidation decreased significantly in normal and hypertrophic cardiomyocytes, there was no difference between normal and hypertrophic cardiomyocytes. But the alterations of fatty acid oxidation after reoxygenation were different between normal and hypertrophic cardiac cells, namely, the fatty acid oxidation of normal cardiomyocytes were activated slowly and slightly, while the rate of fatty acid oxidation of hypertrophic cardiomyocytes increased markedly at the early stage of reoxygenation, and increased further at 8 h of reoxygenation. (5) The rate of apoptosis in hypertrophic cardiocytes increased obviously after hypoxia for 8 h, and increased further and markedly at the early stage of reoxygenation, then gradually decreased to normal level. (6) Dicholoroacetate could inhibit apoptosis of hypertrophic cardiocytes through increasing glucose oxidation and inhibiting the activation of glycolysis and fatty acid oxidation of hypertrophic cardiomyocytes induced by hypoxia-reoxygenation. These data demonstrate that apoptosis in hypertrophic cardiomyocytes after hypoxia-reoxygenation is mainly due to the inhibition of glucose oxidation and the activation of glucolysis and fatty acid oxidation. Furthermore, increasing glucose oxidation may be a new pharmacotherapeutic target to inhibit apoptosis of hypertrophic cardiac cells. PMID- 16220205 TI - Inhibitory effect of rhynchophylline on human ether-a-go-go related gene channel. AB - We studied the effects of Chinese traditional medicine rhynchophylline (Rhy) on human ether-a-go-go related gene (HERG) channel and characterized the electrophysiological properties of Rhy's pharmacological effect on HERG channel using Xenopus oocytes. Xenopus oocytes were injected with either 23 nl (5.75 ng) HERG cRNA or 23 nl distilled water. Xenopus oocytes were randomly assigned to receive one of the following different concentrations of Rhy: (1) control, (2)10 mumol/L Rhy, (3)100 mumol/L Rhy, (4) 500 mumol/L Rhy, (5) 1 000 mumol/L Rhy, (6) 10 000 mumol/L Rhy. Cell currents were recorded in oocytes. The peak tail currents of HERG channel were inhibited by Rhy. The inhibition was in a dose dependent manner [IC(50)=(773.4 +/- 42.5) mumol/L]. Experiment with 100 mumol/L Rhy indicated that the degree of HERG blockade showed some voltage dependence (within -40 mV to -20 mV ). Kinetic analyses revealed that Rhy decreased the rate of channel activation. The findings indicate that Rhy inhibits HERG encoded potassium channels. It may underline the molecular mechanism of myocardial electrophysiological characteristics associated with this drug. PMID- 16220204 TI - [Identification of up-regulated genes induced by angiotensin II in cardiac fibroblasts]. AB - To identify up-regulated genes in adult rat cardiac fibroblasts (CF) induced by angiotensin II (Ang II), suppression subtractive hybridization (SSH) was performed between the CF stimulated by Ang II (tester) and unstimulated CF (driver) to generate subtractive cDNA library. The library was screened with dot blots hybridization to further verify the differentially expressed cDNA clones. Partial positive clones (19 up-regulated genes) were sequenced and BLAST analyzed. Twelve up-regulated genes related to extracellular matrix, cell cycle, intracellular signal transduction, cell cytoskeleton, cell metabolism and 7 new expressed sequence tags (EST) were acquired (GenBank accession number: CN382808, CN382809, CN382810, CN382811, CN382812, CN382813, CN382814). Our data reveal that SSH is a powerful technique of high sensitivity for the detection and cloning of up-regulated genes expressed in CF induced by Ang II, which may be helpful to clarify the mechanism of cardiac remodeling. PMID- 16220206 TI - [No influence of increased frequency on fatigability of tetanic contraction in rat atrophic soleus]. AB - The present study was performed to observe the time course and features of intermittent tetanic contractile function changes in soleus and extensor digitorum longus (EDL) muscles of tail-suspended rats. The optimal stimulating frequency, fatigability and time-dependent recovery after fatigue were measured in isolated muscle strips. The optimal stimulating frequency of soleus and EDL was 60 Hz and 120 Hz in control rats, respectively. It was not changed in 1-week unloaded soleus, but shifted to 80 Hz and 100 Hz in 2- and 4-week unloaded soleus, respectively. The maximal isometric tension (P(o)) of tetanic contraction at optimal stimulating frequency did not alter in 1- and 2-week unloaded soleus, but significantly decreased in 4-week unloaded soleus. After 5 min of fatigue, tetanic contractile tension of control soleus was decreased to 22.8% P(o), but significantly decreased to 10.4%, 10.0% and 11.6% P(o) in 1-, 2- and 4-week unloaded soleus, respectively. The tetanic contractile tension recovered to 98% P(o) in control soleus at the twentieth minute after fatigue, but only recovered to 79.0%, 83.6% and 78.5% P(o) in 1-, 2- and 4-week unloaded soleus. The optimal stimulating frequency, P(o), fatigability and time-dependent recovery of intermittent tetanic contraction were not altered in unloaded EDL compared with control. These results indicate that higher stimulating frequency can compensate the P(o) reduction in 1- and 2-week unloaded soleus, but not in 4-week unloaded soleus. The unloaded soleus, but not EDL, is more susceptible to fatigue than the synchronous controls. The unloaded soleus not only fatigues to a greater extent but also recovers significantly less than the control. PMID- 16220207 TI - CT enteroclysis: technique and clinical applications. AB - CT enteroclysis (CTE) has been gradually evolving with technical developments of spiral and multidetector row CT technology. It has nowadays become a well-defined imaging modality for the evaluation of various small bowel disorders. Volume challenge of 2L of enteral contrast agent administrated to the small bowel via a nasojejunal catheter ensures luminal distension, the prerequisite for the detection of mural abnormalities, also facilitating the accurate visualization of intraluminal lesions. CT acquisition is centered on small bowel loops, reconstructed in thin axial slices and completed by multiplanar views. Image analysis is essentially done in cine-mode on work-stations. CTE is of particular diagnostic value in intermediate or advanced stages of Cohn's disease, including the depiction of extraintestinal complications. It has become the imaging modality of choice for the localization and characterization of small bowel tumors. The cause and degree of low-grade small bowel obstruction is more readily analyzed with the technique of CTE than conventional CT. Limitations of CTE concern the assessment of pure intestinal motility disorders, superficial mucosal lesions and arteriovenous malformations of the small bowel, which are not consistently visualized. CTE should be selectively used to answer specific questions of the small bowel. It essentially contributes to the diagnostic quality of modern small bowel imaging, and therefore deserves an established, well-defined place among the other available techniques. PMID- 16220208 TI - Multislice spiral computed tomography to determine the effects of a recruitment maneuver in experimental lung injury. AB - Although recruitment of atelectatic lung is a common aim in acute respiratory distress syndrome (ARDS), the effects of a recruitment maneuver have not been assessed quantitatively. By multislice spiral CT (MSCT), we analyzed the changes in lung volumes calculated from the changes in the CT values of hyperinflated (V(HYP)), normally (V(NORM)), poorly (V(POOR)) and nonaerated (V(NON)) lung in eight mechanically ventilated pigs with saline lavage-induced acute lung injury before and after a recruitment maneuver. This was compared to single slice analysis near the diaphragm. The increase in aerated lung was mainly for V(POOR) and the less in V(NORM). Total lung volume and intrathoracic gas increased. No differences were found for tidal volumes measured by spirometry or determined by CT. The inspiratory-expiratory volume differences were not different after the recruitment maneuver in V(NON) (from 62+/-18 ml to 43+/-26 ml, P = 0.114), and in V(NORM) (from 216+/-51 ml to 251+/-37 ml, P = 0.102). Single slice analysis significantly underestimated the increase in normally and poorly aerated lung. Quantitative analysis of lung volumes by whole lung MSCT revealed the increase of poorly aerated lung as the main mechanism of a standard recruitment maneuver. MSCT can provide additional information as compared to single slice CT. PMID- 16220209 TI - Brainstem and cerebellar changes after cerebrovascular accidents: magnetic resonance imaging. AB - We illustrate the various types of secondary degeneration in the brainstem and/or cerebellum detected on magnetic resonance (MR) images obtained after cerebrovascular accidents. The changes include: (a) ipsilateral nigral degeneration after striatal infarction; (b) Wallerian degeneration of the pyramidal tract in the brainstem after supratentorial pyramidal tract or motor cortex injury; (c) Wallerian degeneration of the corticopontine tract in the brainstem after frontal lobe infarction; (d) ipsilateral brainstem atrophy and crossed cerebellar atrophy due to an extensive supratentorial lesion; (e) ipsilateral superior cerebellar peduncle atrophy, contralateral rubral degeneration, contralateral inferior olivary degeneration and ipsilateral cerebellar atrophy after dentate nucleus hemorrhage; (f) ipsilateral inferior olivary degeneration after pontine tegmentum hemorrhage; (g) bilateral wallerian degeneration of the pontocerebellar tracts after ventromedial pontine infarction or basis pontis hemorrhage; and (h) ipsilateral cerebellar atrophy after middle cerebellar peduncle hemorrhage. PMID- 16220211 TI - Association of mannose-binding lectin gene (MBL2) polymorphisms with rheumatoid arthritis in an Indian cohort of case-control samples. AB - Single nucleotide polymorphisms in the mannose-binding lectin (MBL2) gene, as well as the serum MBL2 level, have been associated with various autoimmune diseases. We investigated whether such polymorphisms and/or the serum MBL2 level were associated with rheumatoid arthritis (RA) in an Indian population. The frequency of the B variant (codon 54) of the MBL2 gene was quite frequent in the healthy Indian population and was significantly (P=6.35x10(-6)) lower in RA patients. We replicated this association (P=1.78x10(-5)) in an independent cohort of control individuals. Promoter polymorphism at -550 nt showed a significant overrepresentation (P=0.003) of the minor allele G in severe RA patients compared with the less severe group. Haplotype LYA frequency was significantly (P=0.03) high in the less severe group, while the frequency of the HYA haplotype was significantly (P=0.04) increased in the severe RA patients. No statistically significant difference in serum MBL2 was observed as a whole, but the individuals homozygous for the LYA haplotype had significantly lower (P=0.017) serum MBL2 levels compared with individuals homozygous for the HYA haplotype. Therefore, the B variant of the MBL2 gene may be associated with protection from RA in our study population, and the promoter polymorphism (-550 nt) seems to have some role in disease progression. PMID- 16220210 TI - Three-dimensional dynamic magnetic resonance angiography for the evaluation of radiosurgically treated cerebral arteriovenous malformations. AB - We assessed the value of three-dimensional (3D) dynamic magnetic resonance angiography (MRA) for the follow-up of patients with radiosurgically treated cerebral arteriovenous malformations (AVMs). Fifty-four patients with cerebral AVMs treated by radiosurgery (RS) were monitored using conventional catheter angiography (CCA) and 3D dynamic MRA with sensitivity encoding based on the parallel imaging. Cerebral AVM was qualitatively classified by two radiologists into one of five categories in terms of residual nidus size and persistence of early draining vein (I, >6 cm; II, 3-6 cm; III, <3 cm; IV, isolated early draining vein; V, complete obliteration). 3D MRA findings showed a good agreement with CCA in 40 cases (kappa=0.62). Of 23 nidus detected on CCA, 3D dynamic MRA showed 14 residual nidus. Of 28 occluded nidus on 3D dynamic MRA, 22 nidus were occluded on CCA. The sensitivity and specificity of 3D dynamic MRA for the detection of residual AVM were 81% and 100%. 3D dynamic MRA after RS may therefore be useful in association with MRI and can be repeated as long as opacification of the nidus or early venous drainage persists, one CCA remaining indispensable to affirm the complete occlusion at the end of follow-up. PMID- 16220212 TI - A pilot validation study of a new measure of activity in psychosis. AB - BACKGROUND: Wing and Brown [Wing JK and Brown GW (1970) Institutionalism and schizophrenia: a comparative study of three mental health hospitals 1960-1968. Cambridge University Press, London] demonstrated a clear relationship between activity and clinical improvement, using time budget methodology with people with psychosis. However, existing time budget measures are demanding to complete, and simpler, check-box measures of activity rely on subjective frequency judgements and do not include the full range of activities in which an individual might be involved. We report on a pilot validation of a simplified time budget measure of activity levels for routine use as a measure of change with people with psychosis. METHODS: Forty-two participants living in the local community with a schizophrenia spectrum diagnosis were grouped according to length of illness and, within the longer duration group, into high/low activity. All completed the time budget. On a second occasion, 15 participants also completed the subscales of the Social Functioning Scale (SFS) (Br J Psychiatry 157:853-859, 1990) to assess construct validity, and 15 completed the time budget to assess test-retest reliability. RESULTS: The time budget discriminated between duration and activity level groups and showed good inter-rater reliability and test-retest reliability. On the SFS, correlations with subscales measuring withdrawal, activities of daily living and employment were found. CONCLUSIONS: Results indicate that our measure is tapping the activity component of social functioning. A larger scale validation study and investigation of sensitivity to change is underway. PMID- 16220213 TI - Symptom control in children and adolescents with attention-deficit/hyperactivity disorder on switching from immediate-release MPH to OROS MPH Results of a 3-week open-label study. AB - AIM: The aim of this study was to assess the impact of switching from immediate release (IR) methylphenidate (MPH) to OROS MPH (CONCERTA, a once-daily long acting MPH formulation, in children and adolescents with attention deficit/hyperactivity disorder (ADHD). METHODS: Subjects with ADHD aged 6-16 (n=105),who were stably maintained on their current IR MPH regimen (10-60 mg/day), were switched to 18, 36 or 54 mg OROS MPH once daily for 21 days, depending on pre-study daily MPH dose. ADHD symptoms were assessed by parents, teachers and investigators. RESULTS: By Day 21, parent/caregiver IOWA Conners ratings had decreased from baseline by 2.7 points to 5.2 (I/O), and by 1.8 points to 5.0 (O/D). Teacher IOWA Conners ratings were maintained. Decreases in IOWA Conners ratings are indicative of ADHD symptom improvement. Approximately 75% of parents and investigators rated therapy as good or excellent. OROS MPH therapy was well tolerated. CONCLUSIONS: Switching from IR MPH to OROS MPH maintained and may have improved symptom control in children and adolescents with ADHD, during the course of this study. The changes in parent/caregiver IOWA Conners ratings suggest that OROS MPH improves symptom control in the after-school period. This is consistent with the 12-h duration of action previously demonstrated for OROS MPH. PMID- 16220215 TI - Children and adolescents with Chronic Fatigue Syndrome in non-specialist settings: beliefs, functional impairment and psychiatric disturbance. AB - BACKGROUND: Adolescents with Chronic Fatigue Syndrome (CFS) seen in specialist centres have substantial psychological and functional impairment. Beliefs about activity levels may be important in the development of CFS. METHOD: The aim was to investigate psychological and functional impairment, and beliefs in children and adolescents with CFS recruited from non-specialist services. A total of 30 such individuals participated, and 30 young people with Inflammatory Bowel Disease (IBD) formed the comparison group. RESULTS: Emotional symptoms and disorder were high in both groups. In all, 50% of those with CFS and 30% with IBD reached the threshold for emotional disorder according to the Strengths and Difficulties Questionnaire (SDQ) parent report, although this difference did not reach statistical significance. Participants with CFS scored statistically significantly higher on measures of functional impairment, including school non attendance, compared to those with IBD. According to questionnaire responses, those with CFS were statistically significantly more likely to favour rest rather than exercise compared to those with IBD. Comparison of parental beliefs did not show such a difference. CONCLUSIONS: These young people with CFS were at high risk of psychiatric disorder. They were substantially disabled when compared to individuals with a known chronic illness. Also, as a group, they were characterised by a preference for rest rather than exercise. PMID- 16220214 TI - 12-month efficacy and safety of OROS MPH in children and adolescents with attention-deficit/hyperactivity disorder switched from MPH. AB - PURPOSE: The aim of this study was to evaluate long-term clinical treatment with OROS methylphenidate (MPH) (Concerta) in children and adolescents with attention deficit/hyperactivity disorder (ADHD) who had been previously treated with immediate release (IR) MPH. METHODS: Subjects aged 6-16 years (n=105) who were stable on IR MPH (10-60 mg/day) were switched to 18, 36 or 54 mg OROS MPH once daily for 21 days, depending on prestudy MPH dose. Subjects who benefited from OROS MPH could continue in a 12-month extension period. ADHD symptoms and treatment response were assessed by parents/caregivers and investigators. RESULTS: Out of 105 enrolled children, 101 completed the 21-day treatment phase. In all, 89 parents/caregivers (88.1%) wanted their child to continue with the study treatment into the extension phase, and 56 children (63 %) completed the 1 year trial. The parent/caregiver global assessment of satisfaction ranged from 49 to 69% during the extension phase, and 49 to 71% of investigators rated the treatment as adequate. Efficacy and satisfaction were found more commonly in patients in the older age group (10-16 years), those on a higher dose (36 mg or 54 mg) and with the predominantly inattentive ADHD subtype. OROS MPH was well tolerated. CONCLUSIONS: Children and adolescents can effectively and safely be switched from IR MPH to OROS MPH with improved symptom control and compliance. PMID- 16220216 TI - Epidemiology and phenomenology of obsessive-compulsive disorder in non-referred young adolescents: a Polish perspective. AB - OBJECTIVE: The aim of this study was to investigate the frequency and phenomenology of obsessive-compulsive disorder (OCD) and subclinical OCD in a non referred population of young Polish adolescents. METHOD: A two stage ascertainment procedure (school screening and diagnostic evaluation) was used to identify affected individuals. In the first stage, 3,100 pupils were asked to complete the Polish version of the 20-item Leyton Obsessional Inventory-Child Version (LOI-CV). In the diagnostic stage, the presence of obsessions and compulsions was assessed with the author's structured interview questionnaire based on DSM-IV and ICD-10 diagnostic criteria for OCD; the Polish version of the Children's Yale-Brown Obsessive-Compulsive Scale (CY-BOCS) was used to rate the symptom severity. RESULTS: A frequency of 0.38 % was found for OCD and 2.7 % for subclinical OCD. There was no significant difference in the phenomenology, demographic characteristic or socio-familial variables comparing the diagnosed OCD and subclinical OCD subjects. CONCLUSIONS: The identified OCD cases had characteristics similar to those of previously described clinical and non referred samples. PMID- 16220217 TI - The Children's Depression Inventory and classification of major depressive disorder: validity and reliability of the Danish version. AB - The study examines the validity and reliability of the Danish version of the Children's Depression Inventory (CDI) in a child psychiatric population. Participants were 149 child psychiatric patients aged 8-13 and their parents. After diagnostic interview with the Kiddie-Schedule for Affective Disorders and Schizophrenia, the children completed the CDI. A subgroup of 44 children repeated the CDI after 2 weeks. The psychometric properties of the Danish CDI were similar to those reported for the English version. CDI is moderately correlated with other measures for depressive disorder, but the instrument is not sufficiently reliable or valid to be used as a single diagnostic or screening measure in a child psychiatric population. PMID- 16220218 TI - DSM-IV or ICD-10-DCR diagnoses in child and adolescent psychiatry: does it matter? AB - OBJECTIVE: DSM-IV is the most widely used diagnostic classification system in research, whereas ICD-10 is more widely used clinically. Knowledge of differences is essential when research findings are implemented in daily clinical practice. We examined differences between the two diagnostic systems regarding three major child psychiatric diagnostic categories. METHODS: A total of 199 consecutively referred, child psychiatric patients were interviewed with a semistructured diagnostic interview (K-SADS-PL) including questions covering specific ICD-10-DCR criteria, and diagnosed according to both diagnostic systems. RESULTS: Differences were found regarding the diagnoses major depressive disorder/depressive episode and attention deficit hyperactivity disorder/disturbance of activity and attention. In both cases, more children met DSM-IV-TR criteria than ICD-10-DCR criteria. The diagnosis, oppositional defiant disorder, proved interchangeable between the two diagnostic systems. CONCLUSION: Differences between diagnostic systems must be taken into account when research findings using one diagnostic system are implemented with children diagnosed by another diagnostic system. PMID- 16220219 TI - Early-onset schizophrenia: a 15-year follow-up. AB - The study describes the psychopathological and social outcome of patients treated for schizophrenia in adolescence (mean age at onset 16.0 years/SD 1.52) after a mean follow-up period of 15.4 years (10.2-21.2 years). Out of 55 patients consecutively admitted to hospital, 47 (85 %) could be traced and 39 (71 %) could be re-examined. At follow-up, 33/39 patients (85 %) had had at least one readmission. Full remission of global psychopathological symptoms [Clinical Global Impression (CGI) or = 85 years 35 new cases per 1,000 occur every year. Atrial fibrillation is associated with an increased mortality. CLASSIFICATION: Atrial fibrillation is classified according to criteria of time: acute, paroxysmal, persistent, or permanent. On this basis this is essential for decisions about the optimal time for cardioversion and especially with respect to duration of anticoagulation before and after the intervention. A classification according to concomitant diseases (risk factors for thromboembolic complications) defines patients who should receive an oral anticoagulation. THERAPY: Patients with lone (idiopathic) atrial fibrillation 60 years of age do not need oral anticoagulation. A cardioversion with class I antiarrhythmic drugs can only be considered for patients with acute atrial fibrillation without further heart diseases. For prophylaxis after cardioversion amiodarone with its high efficiency and its low proarrhythmic risk is the first choice. Still, this has to be done with caution and regarding the various extracardial side effects. For all procedures to restore sinus rhythm it has to be kept in mind that there is no evidence yet that they improve the prognosis. Therefore, the concept of frequency control under long-term anticoagulation is more and more favored. COMPLICATIONS: The most severe complications of atrial fibrillation are thromboembolic diseases, especially strokes. The risk for strokes can be reduced by nearly 70% by oral anticoagulation in patients with non rheumatic atrial fibrillation. The risk of bleeding complications, mostly extracranial, has to be weighed against the primary goal of anticoagulation. Bleeding complications closely correlate with the intensity of anticoagulation. In patients with nonrheumatic atrial fibrillation an INR between 2.0 and 3.0 has found to be effective and safe. Above an INR level of 4.0 the bleeding risk is much higher without an additional protective effect. PMID- 16220253 TI - [Hepatocyte transplantation]. AB - BACKGROUND: Transplantation of hepatocytes is considered a promising technology for the cell therapy of liver diseases. Cells are isolated from donor livers, which are not allocated for organ transplantation and transplanted into the liver of a suitable recipient. Ideally, the transplanted cells functionally replace the hepatocytes of the diseased organ and restore its metabolic capacity either permanently or for a period of bridging to organ transplantation. METHODS AND RESULTS: Although about 50 cases of clinical hepatocyte transplantation have been documented, therapeutic benefit is doubtful, and the reasons for this are largely unknown. Minor quality of the transplanted cells isolated from marginal donor livers may be a major cause. Therefore, animal models have been established, which enable scientists to develop novel procedures of hepatocyte transplantation and to specifically study the mechanisms of hepatocyte integration in the host liver. Hopefully, results generated in animal models will set the basis for the design of novel procedures of hepatocyte transplantation individualized to the underlying disease in order to provide a growth advantage for donor over host cells. PERSPECTIVES: The plasticity of stem cells and their proliferative potential is the basis for current efforts to generate stem cell-derived hepatocytes of transplantation quality for the treatment of liver diseases. PMID- 16220254 TI - [The actual role of general practice in the dutch health-care system. Results of the second dutch national survey of general practice]. AB - A second Dutch National Survey of General Practice was carried out in 2001 with the aim of providing actual information about the role of general practice in the Dutch health-care system for researchers and policy makers. Data were collected on different levels (patients, general practitioners, practices) and included morbidity (self-report and presented to general practitioners), diagnostic and therapeutic interventions, doctor-patient communication, and background characteristics. Compared to 1987 (the first National Survey), Dutch general practitioners had organized their work more efficiently. Patients were less satisfied (78% satisfied) about the organizational aspects of general practice care than about the care actually provided (90% satisfied). Dutch general practitioners provide high-quality care: on average, their performance was in 74% of cases in accordance with national guidelines. Communication in general practice had become less social and more medically oriented compared to 1987. General practice still acts in a gatekeeper role; this is illustrated by 96% of contacts handled solely by the general practitioner. PMID- 16220255 TI - [Erythrocyturia and proteinuria. When should a nephrologist be consulted?]. PMID- 16220256 TI - [Polycythemia vera and D-dimer-negative thromboembolism]. AB - CASE REPORT: A 65-year-old patient with polycythemia vera (PV) was admitted with a painful edema of the right arm lasting for 24 h. The D-dimer assay was negative. By phlebography the patient was diagnosed with a fresh thrombosis of the right subclavian vein. 1 week later she developed a D-dimer-negative symptomatic pulmonary embolism. CONCLUSION: If clinical signs of a thromboembolic event are present, the negative predictive value of the D-dimer assay is not sufficient to abandon further definitive diagnosis. Since thromboembolic events are frequent in PV patients, a prophylactic treatment with low-dose acetylsalicylic acid is recommended. PMID- 16220257 TI - [Rapid healing of a therapy-refractory diabetic foot after transplantation of autologous bone marrow stem cells]. AB - BACKGROUND: The diabetic foot mainly depends on painless pressure lesions, which are based on diabetic polyneuropathy and microangiopathy. In these cases the regenerative potential of adult autologous mononuclear stem cells could serve as causal therapy. HISTORY AND CLINICAL FINDINGS: A 63-year-old patient with long lasting type 2 diabetes mellitus suffers from a reduced walking distance of 200 m and a therapy-refractory ulcer at the right ball of the great toe. Therefore, the authors have decided to perform a combined intraarterial and intramuscular transplantation of stem cells into the right limb for the first time on this disease. THERAPY AND RESULTS: After harvesting of bone marrow the mononuclear cell fraction was separated (157 x 10(6) cells). Thereafter, the fractional intraarterial and intramuscular transplantation of the cell suspension was performed (10 ml each). Already 8 weeks later, the ulcer healed completely, after 6 months the walking distance increased by > 100%, on venous occlusion plethysmography the arterial blood circulation at rest increased by 23% and the reactive hyperemia by 56%. CONCLUSION: The combined intraarterial and intramuscular transplantation of autologous bone marrow stem cells could constitute a novel, clinically feasible and safe therapy for patients with diabetic foot syndrome. The success of this approach may be ascribed to microangiogenesis and to an anti-inflammatory effect of the transplanted stem cells. PMID- 16220258 TI - [Acute and chronic pain]. PMID- 16220259 TI - [Splenic infarct in a patient with HIV infection]. PMID- 16220261 TI - [Method and design of the Telephone Health Survey 2003]. AB - Telephone surveys are increasingly being used in public health research, also in Germany. In 2003 the Robert Koch Institute completed the first nationwide telephone survey predominantly on conditions and prevalence of selected chronic diseases (GSTel03). Its findings are presented in this publication. The tasks of planning, implementation and realization of procedures used in telephone surveys differ in many aspects from other modes of data collection. This article deals with the most important methodological aspects that were implemented and used in the course of the GSTel03 to ensure a high level of data quality. These include sample design, questionnaire development and the process of data collection (including the recruitment and training of the staff). Finally steps of data preparation for analysis including weighting procedures are described and calculation of contact and cooperation rates are reported. PMID- 16220262 TI - Application of computer imaging, stripping voltammetry and mass spectrometry to study the effect of lead (Pb-EDTA) on the growth and viability of early somatic embryos of Norway spruce (Picea abies /L./ Karst.). AB - Image analysis (IA) was used to determine the areas and circumferences of clusters of early somatic embryos (ESEs) of the Norway spruce (Picea abies /L./Karst.). Results obtained from IA were compared with the fresh weights of the ESE clusters and their esterase activities. The areas of the ESE clusters correlated well with both the increases in fresh weight (R2=0.99) of the ESEs and their esterase activities (R2=0.99). In addition, we studied the viability of the ESEs, which was determined by (a) double staining with fluorescein diacetate and propidium iodide (the resulting fluorescence was quantified by IA) and (b) determining esterase activity using a spectrofluorimetric detector. The results obtained with IA and esterase assay were comparable (the deviation between the tangents of the bisectors was 6.4%). IA was also used to study the effect of Pb EDTA chelate (50, 250 and 500 microM) on the viability of the ESEs and on the growth of clusters. The presence of Pb-EDTA markedly slowed the growth of ESEs clusters (by more than 65% with 250 microM of Pb-EDTA after 288 h of cultivation) and decreased the viability of ESEs (by more than 30% with 500 microM of Pb-EDTA after 288 h of cultivation). The lead concentration in the ESEs was determined by differential pulse anodic stripping voltammetry and increased with the external lead concentration and the time of treatment from 100 to 600 pg Pb/100 mg of fresh weight of ESEs. Glutathione is a diagnostic marker of the influence of Pb EDTA on ESEs and its content was determined by high-performance liquid chromatography coupled with mass spectrometry. The glutathione content changed linearly with treatment time and the applied external lead concentration. The highest glutathione content was obtained at 250 microM of Pb-EDTA after 192 h of cultivation. PMID- 16220263 TI - Production of a lipolytic enzyme originating from Bacillus halodurans LBB2 in the methylotrophic yeast Pichia pastoris. AB - A gene encoding a lipolytic enzyme amplified from the alkaliphilic bacterium Bacillus halodurans LBB2 was cloned into the pPICZalphaB vector and integrated into the genome of the protease deficient yeast strain Pichia pastoris SMD1168H. This previously undescribed enzyme was produced in active form, and cloning in frame with the Saccharomyces cerevisiae secretion signal (alpha-factor) enabled extracellular accumulation of correctly processed enzyme, with an apparent molecular mass of 30 kDa. In shake-flask cultivations, very low production levels were obtained, but these were significantly improved by use of a "batch-induced" cultivation technique which allowed a maximum enzyme activity of 14,000 U/l using p-nitrophenyl butyrate (C-4) as a substrate and a final extracellular lipolytic enzyme concentration of approximately 0.2 g/l. Partial characterization of the produced enzyme (at pH 9) revealed a preference for the short-chain ester (C-4) and significant but lower activity towards medium (C5-C6) and long (C16 and C18) fatty acid chain-length esters. In addition, the enzyme exhibited true lipase activity (7,300 U/l) using olive oil as substrate and significant levels of phospholipase activity (6,400 U/l) by use of a phosphatidylcholine substrate, but no lysophospholipase activity was detected using a lysophosphatidylcholine substrate. PMID- 16220264 TI - Miple1 and miple2 encode a family of MK/PTN homologues in Drosophila melanogaster. AB - Midkine (MK) and Pleiotrophin (PTN) are small heparin-binding cytokines with closely related structures. To date, this family of proteins has been implicated in multiple processes, such as growth, survival, and migration of various cells, and has roles in neurogenesis and epithelial-mesenchymal interaction during organogenesis. In this report, we have characterized two members of the MK/PTN family of proteins in Drosophila, named Miple1 and Miple2, from Midkine and Pleiotrophin. Drosophila miple1 and miple2 encode secreted proteins which are expressed in spatially restricted, nonoverlapping patterns during embryogenesis. Expression of miple1 can be found at high levels in the central nervous system, while miple2 is strongly expressed in the developing midgut endoderm. The identification of homologues of the MK/PTN family in this genetically tractable model organism should allow an analysis of their function during complex developmental processes. PMID- 16220265 TI - Vertebrate neurogenin evolution: long-term maintenance of redundant duplicates. AB - The majority of the cranial sensory neurons of vertebrates, including all of those concerned with the special senses of hearing, balance and taste, are derived from the neurogenic placodes. A number of studies have shown that the production of neuronal cells by the placodes is dependent upon the function of the neurogenin (ngn) gene family of basic helix-loop-helix transcription factors. One member of the gene family is expressed in each placode, suggesting that this specificity of expression could help to determine different neuronal classes. An interesting feature of this expression, however, is that the expression patterns vary amongst the vertebrates; for example, mammals and fish express ngn-1 in the ophthalmic trigeminal placode where birds use ngn-2. This prompted us to use phylogenetic and genomic analysis to unravel the evolutionary history of this gene family. We determined that the duplication that created the neurogenin-1 and -2 subfamilies occurred deep in the vertebrate lineage before the divergence of bony fish 450 million years ago and suggest that concurrent expression of both genes was probably maintained in all neurogenic placodes until after the divergence of birds and mammals 270 million years ago. PMID- 16220266 TI - Kinetic study of the toxicity of zinc and lead ions to the heavy metals accumulating fungus Paecilomyces marquandii. AB - Studies have been carried out to determine the toxicity of zinc and lead ions to germinating spores and hyphal growth of heavy metal accumulating fungus Paecilomyces marquandii (former Verticillium marquandii). Inhibitive concentration (IC50) of zinc and lead ions was assayed by three different methods: image analysis, nephelometric on-line measurement and microcalorimetry. A kinetic model of spore germination and germ tube elongation was formulated and used as an auxiliary tool to determine IC50 values upon image analysis data. The inhibitive effect of Zn2+ and Pb2+ to P. marquandii spores was mathematically described by the Edwards equation. Comparing the obtained IC50 values, lead ions occurred to be more toxic to the germinating spores of P. marquandii than zinc ions (2.80 and 5.20 mM, respectively), although zinc ions induced a more significant delay in the development of the hyphae (13.84 h for 5 mM of Zn2+ and 9.30 h for 5 mM of Pb2+), which was demonstrated by the lengthened lag-phase (spore-swelling phase). PMID- 16220267 TI - Biosoftening of coir fiber using selected microorganisms. AB - Coir fiber belongs to the group of hard structural fibers obtained from coconut husk. As lignin is the main constituent of coir responsible for its stiffness, microbes that selectively remove lignin without loss of appreciable amounts of cellulose are extremely attractive in biosoftening. Five isolated strains were compared with known strains of bacteria and fungi. The raw fiber treated with Pseudomonas putida and Phanerocheate chrysosporium produced better softened fiber at 30+/-2 degrees C and neutral pH. FeSO4 and humic acid were found to be the best inducers for P. chrysosporium and P. putida, respectively, while sucrose and dextrose were the best C-sources for both. Biosoftening of unretted coir fibers was more advantageous than the retted fibers. Unlike the weak chemically softened fiber, microbial treatment produced soft, whiter fibers having better tensile strength and elongation (44.6-44.8%) properties. Scanning electron microscopy photos showed the mycelia penetrating the pores of the fiber, removing the tylose plug and degrading lignin. PMID- 16220268 TI - Ewing's sarcoma presenting as a solitary cyst. AB - This case describes a 10-year-old girl who developed a Ewing's sarcoma in her proximal fibula. The radiologic features mimicked those of a unicameral bone cyst. The presence of pain and the atypical location led to a prompt biopsy and the correct diagnosis. The mechanism of this unusual radiographic presentation is discussed. PMID- 16220271 TI - Intramedullary insertion of the patellar tendon. AB - This case represents a rare presentation of an intramedullary insertion of the patellar tendon in an otherwise healthy 8-year-old girl. To our knowledge no such phenomenon has previously been reported in the English literature. PMID- 16220269 TI - Synovial sarcoma with radiological appearances of primitive neuroectodermal tumour/Ewing sarcoma: differentiation by molecular genetic studies. AB - Synovial sarcoma (SS) arises in soft tissues but may invade adjacent bone. We describe a case of SS presenting as aggressive lysis of the proximal ulna, the imaging of which suggested a primary bone lesion. Needle biopsy showed a "small round blue cell tumour", and a primitive neuroectodermal tumour (PNET)/Ewing sarcoma was suggested on the basis of the imaging appearances. The definitive diagnosis of synovial sarcoma was made following molecular genetic studies, which demonstrated a fusion product incorporating the genes SYT and SSX1. The importance of correct diagnosis to guide appropriate management, and, therefore, the necessity for molecular genetic studies, is discussed. PMID- 16220272 TI - "Dropped-head" syndrome due to isolated myositis of neck extensor muscles: MRI findings. AB - MRI findings of a patient with dropped-head syndrome due to focal myositis of the neck extensor muscles are presented. MRI showed oedematous changes and marked enhancement of the neck extensor muscles. After therapy MRI demonstrated disappearance of the abnormal findings. PMID- 16220270 TI - MRI features of extraskeletal myxoid chondrosarcoma. AB - OBJECTIVE: To describe the MRI features of extraskeletal myxoid chondrosarcoma in comparison with clinicopathologic findings. DESIGN AND PATIENTS: The study comprised 12 male subjects and seven female subjects with a mean age of 53 years (range 16-76 years). MRI findings, evaluated by two radiologists with agreement by consensus, were compared for histopathologic features. RESULTS: The tumor size ranged from 2.0 cm to 20.0 cm (mean 8.9 cm). Fusion gene transcripts could be detected in 13 (68%) of the 19 cases: EWS-CHN in nine cases, TAF2N-CHN in three, and TFG-TCH in one. There were six fusion-negative cases. Signal characteristics on T1-weighted and T2-weighted MR images were non-specific with regard to each cytogenetic variant. Peripheral enhancement was seen more frequently in tumors with the EWS-CHN variant than in those with other cytogenetic variants. The characteristic pattern of enhancement corresponded to the presence of fibrous septa and peripheral areas of high cellularity within lobules, by correlation with pathologic findings. All cases with TAF2N-CHN or TFG-TCH variants showed invasion of extracompartmental structure, bone, or vessels. CONCLUSION: Extraskeletal myxoid chondrosarcoma is an uncommon soft-tissue malignancy that may be recognized by MRI features of multi-lobular soft-tissue mass often invading extracompartmental, bony, and vascular structures. PMID- 16220273 TI - Presence and co-localization of vasoactive intestinal polypeptide with neuronal nitric oxide synthase in cells and nerve fibers within guinea pig intrinsic cardiac ganglia and cardiac tissue. AB - The presence of vasoactive intestinal polypeptide (VIP) has been analyzed in fibers and neurons within the guinea pig intrinsic cardiac ganglia and in fibers innervating cardiac tissues. In whole-mount preparations, VIP-immunoreactive (IR) fibers were present in about 70% of the cardiac ganglia. VIP was co-localized with neuronal nitric oxide synthase (nNOS) in fibers innervating the intrinsic ganglia but was not present in fibers immunoreactive for pituitary adenylate cyclase-activating polypeptide, choline acetyltransferase (ChAT), tyrosine hydroxylase, or substance P. A small number of the intrinsic ChAT-IR cardiac ganglia neurons (approximately 3%) exhibited VIP immunoreactivity. These few VIP IR cardiac neurons also exhibited nNOS immunoreactivity. After explant culture for 72 h, the intraganglionic VIP-IR fibers degenerated, indicating that they were axons of neurons located outside the heart. In cardiac tissue sections, VIP IR fibers were present primarily in the atria and in perivascular connective tissue, with the overall abundance being low. VIP-IR fibers were notably sparse in the sinus node and conducting system and generally absent in the ventricular myocardium. Virtually all VIP-IR fibers in tissue sections exhibited immunoreactivity to nNOS. A few VIP-IR fibers, primarily those located within the atrial myocardium, were immunoreactive for both nNOS and ChAT indicating they were derived from intrinsic cardiac neurons. We suggest that, in the guinea pig, the majority of intraganglionic and cardiac tissue VIP-IR fibers originate outside of the heart. These extrinsic VIP-IR fibers are also immunoreactive for nNOS and therefore most likely are a component of the afferent fibers derived from the vagal sensory ganglia. PMID- 16220274 TI - Identification of Pip4k2beta as a mechanical stimulus responsive gene and its expression during musculoskeletal tissue healing. AB - To investigate the mechano-transduction system of cells, we identified genes responsive to a cyclic mechanical stimulus. MC3T3.E1 cells were cultured on a computer-controlled vacuum-pump-operated device designed to provide a cyclic mechanical stimulus. A maximum elongation of 15% of membrane at 10 cycles/min (3 s extension followed by 3 s relax per cycle) was repeated for 48 h. By means of a differential display, the gene expression pattern of cells exposed to the stimulus was compared with that of unexposed cells. As a result, a gene fragment that was exclusively expressed in mechanically stressed cells was identified. By using expressed sequence tag walking together with the oligo-capping method, this gene was identified as phosphatidylinositol 4-phosphate 5-kinase type II beta (initially known as Pip5k2beta but now reclassified as Pip4k2beta). The specific up-regulation of Pip4k2beta upon mechanical stimulus was also confirmed by using another apparatus, viz. a computer-controlled linearized-stepping motor system. To examine the involvement of the cyclic mechanical stimulus in the regulation of Pip4k2beta expression in musculoskeletal tissue, we created an Achilles tendon transection model in rabbits. The temporal expression of Pip4k2beta was assessed by means of a quantitative reverse-transcribed polymerase chain reaction. In the gastrocnemius muscle, expression of Pip4k2beta rapidly decreased 1 week after transection but was restored to normal levels at 4 weeks. In the Achilles tendon, however, expression remained decreased until 4 weeks after transection. We suggest that the expression of Pip4k2beta can be used as a marker for cells receiving a suitable mechanical stimulus. PMID- 16220276 TI - [AIDS 2005: report from the 10th German and 16th Austrian AIDS Congress, 1-4 June 2005, Vienna]. PMID- 16220275 TI - [ASS or clopidogrel plus esomeprazole after bleeding from an ulcer]. PMID- 16220277 TI - Reactive lymphoid hyperplasia one month after LASIK surgery. AB - BACKGROUND: To describe a case of reactive lymphoid hyperplasia following laser assisted in situ keratomileusis (LASIK). METHODS: This observational case report describes a 31-year-old man who underwent LASIK and presented 1 month later with a fleshy, conjunctival (plical) tumour in the left eye. An excisional biopsy of the tumour was performed. RESULTS: Histopathology of the excised tumour revealed reactive lymphoid hyperplasia involving the left conjunctiva. DISCUSSION: Conjunctival lymphomas can masquerade as chronic conjunctivitis and can be preceded by reactive lymphoid hyperplasia. It is important to identify and differentiate these tumours. This report describes the unusual occurrence of a lymphoid conjunctival tumour after LASIK eye surgery. PMID- 16220278 TI - Disclosing disease mechanisms with a spatio-temporal summation paradigm. AB - BACKGROUND: We develop the logic for a stimulus that can evaluate cone-dependent spatial summation and detail the modelling and interpretation of thresholds obtained with this stimulus. METHODS: Fifteen observers participated, including two young normals tested extensively in control experiments, and a clinical trial based on four observers with age-related macular degeneration (AMD), four age similar controls and five young observers. Monocular spatial summation functions were measured with contrast-modulated Gabor targets that approximated the optimal visual contrast detector. Thresholds were returned from a yes/no adaptive psychophysical algorithm. By fine titration along the size domain it was demonstrated that the spatial summation of normal observers can be adequately described by a two-component model. A reduced set of variables are proposed for clinical applications and the model was applied to data derived using these variables in persons with AMD and age-similar controls. RESULTS: We do not find a significant age-related loss of contrast sensitivity in our normal group. On the other hand, persons with early AMD exhibited a 0.41 log unit loss of sensitivity (P=0.04) from age-similar controls, without any change in their maximum summation area (A(max)). CONCLUSIONS: The nature of the spatial summation is consistent with the interpretation that early AMD produces a decrease in cone input to post receptoral mechanisms in the absence of neural remodelling. PMID- 16220279 TI - Glaucoma follow-up by the Heidelberg retina tomograph--new graphical analysis of optic disc topography changes. AB - BACKGROUND: The Heidelberg Retina Tomograph (HRT) is a commercially available instrument for the detection of glaucomatous damage by analysis of optic nerve head topography. The main purpose of the study was to investigate the ability of HRT to detect changes in optic disc topography indicating progression of optic neuropathy in eyes with open-angle glaucoma or, in eyes with ocular hypertension, conversion to open-angle glaucoma. METHODS: Fifty-nine subjects (34 with ocular hypertension, 25 with glaucoma) from the glaucoma service at Sahlgrenska University Hospital were included in this study. One eye of each patient was selected. All participants underwent thorough clinical examination, including HRT, high-pass resolution perimetry (HRP), and optic disk photography. After a mean follow-up time of 50 months, patients were re-examined. Based on analyses of optic disc photographs and HRP, eyes were classified into one of two groups: progressive or stable. The differences between baseline and follow-up HRT parameters in the two groups were analysed. The topographic HRT change images were also compared after digital image processing. A pixel ratio was calculated defined as the ratio between the area of pixels representing deepening of the disc surface and the total disc area. Receiver operating characteristic (ROC) curves for HRT parameters and pixel ratio were compared. RESULTS: In the group judged to have progressive optic neuropathy, a statistically significant change between baseline and follow-up examination was found for the following HRT parameters: cup shape measurement, classification index, the third moment in contour, cup/disc ratio, cup area, rim area, and area below reference). In the stable group no HRT parameters had changed significantly. A well-defined distinction between the two groups was found by comparing digitally processed HRT change images. The area under the ROC curve was larger for pixel ratio than for any of the HRT parameters. CONCLUSIONS: The HRT is a useful tool for long-term follow-up of glaucomatous optic neuropathy. Digital image processing of HRT change images could facilitate the detection of progressive change. PMID- 16220280 TI - [The subclavicular route for the pectoralis major myocutaneous flap]. AB - BACKGROUND: The pectoralis major myocutaneous flap (PMMF) is an important reconstructive tool for lesions in the head and neck region. Using the supraclavicular route, the PMMF reliably transfers large amounts of well vascularized skin and muscle into defects of the upper aerodigestive tract. However, limited length and arc of rotation as well as excessive bulk can be problematic. PATIENTS AND METHODS: In the current study, these problems have been addressed by passing the pedicle deeply to the clavicle. Following flap harvest, the pedicle was passed in the subclavicular plane in 15 head and neck cancer patients for primary and secondary reconstruction. RESULTS: Using this route it was possible to increase the medium length of PMMF to 3 cm compared to the supraclavicular route. No total flap necrosis occurred, however, temporary complications were observed in three of 15 cases (20%)--partial flap necrosis occurred in two cases and fistula formation was observed in one case. This rate is in accordance with complication rates described for the supraclavicular route. However, in long-term follow-ups we observed a fracture of the clavicle in two patients in whom, in contrast to the others, the periostium was not only prepared posteriorly but over the whole circumference. CONCLUSIONS: The subclavicular route for PMMF increases the length and arc of rotation available for reconstruction without compromising vascular supply to a higher degree than with the conventional supraclavicular route. Furthermore, this concept decreases the bulk of the PMMF pedicle which is functionally and cosmetically favourable. Thus, the subclavicular route of PMMF is safe and allows an extension of the reconstructive possibilities. PMID- 16220281 TI - [Dysarthria as a cardinal neurological symptom]. PMID- 16220283 TI - [(Central) auditory processing disorders. Remarks on the ASHA Technical Report]. PMID- 16220284 TI - [A multicentre comparative study of the ESPrit and the Nucleus 22]. AB - BACKGROUND: Cochlear implant recipients often report additional difficulty in comprehension of speech in noisy conditions and of softly spoken speech. The aim of this clinical study was to evaluate and compare the performance advantages offered by the ear level ESPrit 3G for experienced Nucleus Mini 22 cochlear implantees. PATIENTS AND METHODS: Twenty-eight German-speaking implanted subjects, who had had experience with either the Spectra 22 or the ESPrit 22 for at least 6 months, were evaluated with their current processor and the ESPrit 3G (on microphone, M, and whisper, W, settings) following a 4-week trial. Freiburger monosyllabic words (FMW) were used at soft and conversational levels in quiet conditions and Oldenburger sentences (OLSA) were used in noisy conditions to compare performance. Subjective impressions of sound quality and user aspects were evaluated and combined with data from 31 English-speaking subjects from a parallel study. RESULTS: In comparison to the previously worn processor, statistically significantly superior performance (p<0.001) was observed at soft and conversational levels in quiet conditions for FMW in 15 subjects when using the W setting and in noisy conditions for the OLSA in 21 subjects when using the M setting (p<0.001). The ESPrit 3G was preferred by 86% of subjects (51/59). CONCLUSION: The ESPrit 3G for Nucleus 22 users has the potential to further improve speech understanding in quiet conditions at soft intensity levels and also in noisy conditions at conversational levels relative to the currently worn speech processor, the Spectra 22 or the ESPrit 22, for the majority of subjects. Subjectively, together with the improvement in sound quality, the majority of subjects also reported improved ease of use and wearer comfort. PMID- 16220282 TI - [Plastic surgery of skin defects in the face. Principles and perspectives]. AB - The difficulties in facial reconstruction derive from the unique character of the face and the availability of local matching tissues. Facial reconstructive surgery must aim at a functionally and aesthetically rehabilitated patient. The performance of facial plastic surgery requires an understanding and the application of many important principles. The aim of this paper is to review the critical factors to be considered in the management of surgical wounds by second intention healing, primary closure, skin grafting, and repair with local or distant free flaps. The key concepts useful in flap choice and implementation are discussed. In addition, an overview of new developments in tissue engineering and gene therapy as they relate to facial plastic surgery is provided. PMID- 16220286 TI - Pliocene large-mammal assemblages from Northern Chad: sampling and ecological structure. AB - Numerous Pliocene large-mammal assemblages have been discovered in Chad over the last decade. They offer a unique opportunity to understand the settings in which important chapters of Hominid evolution took place in Central Africa. However, it is crucial to first investigate both sampling and taxonomic homogeneity for these Chadian assemblages because they occur over large sectors in a sandy desert that offers virtually no stratigraphic section. Using cluster analysis and ordination techniques, we show that the three Pliocene sectors from Chad are homogeneous and adequate sampling units. Previous stable isotope analyses on these assemblages have indicated that the environment became richer in C(4) plants between approximately 5.3 and 3.5-3 Ma. To test whether this environmental change has affected the structure of palaeo-communities, we assigned body mass, trophic and locomotor eco-variables to mammal species from the three sectors. Statistical analysis shows that the overall ecological structure of the assemblages is not linked with the opening of the plant cover, and eco-variables show no temporal trend from the oldest sector to the youngest. For example, there is no significant change in the relative diversity of grazing and browsing taxa, although mixed feeders are less diversified in the youngest sector than in the preceding one. This pattern apparently does not result from potential biases such as methodological artefacts or taphonomic imprint. Instead, it seems that local heterogeneous environmental factors have played a major role in shaping the ecological spectrum of Chadian mammal palaeo-communities during the Pliocene. PMID- 16220285 TI - Molecular mechanisms of optic axon guidance. AB - Axon guidance is one of the critical processes during vertebrate central nervous system (CNS) development. The optic nerve, which contains the axons of retinal ganglion cells, has been used as a powerful model to elucidate some of the mechanisms underlying axon guidance because it is easily manipulated experimentally, and its function is well understood. Recent molecular biology studies have revealed that numerous guidance molecules control the development of the visual pathway. This review introduces the molecular mechanisms involved in each critical step during optic axon guidance. Axonal projections to the optic disc are thought to depend on adhesion molecules and inhibitory extracellular matrices such as chondroitin sulfate. The formation of the head of the optic nerve and the optic chiasm require ligand-receptor interactions between netrin-1 and the deleted in colorectal cancer receptor, and Slit proteins and Robo receptors, respectively. The gradient distributions of ephrin ligands and Eph receptors are essential for correct ipsilateral projections at the optic chiasm and the topographic mapping of axons in the superior colliculus/optic tectum. The precise gradient is regulated by transcription factors determining the retinal dorso-ventral and nasal-temporal polarities. Moreover, the axon guidance activities by Slit and semaphorin 5A require the existence of heparan sulfate, which binds to numerous guidance molecules. Recent discoveries about the molecular mechanisms underlying optic nerve guidance will facilitate progress in CNS developmental biology and axon-regeneration therapy. PMID- 16220287 TI - Reproductive history, occupational exposures, and thyroid cancer risk among women textile workers in Shanghai, China. AB - OBJECTIVES: Thyroid cancer risk has been previously associated with increased age at first pregnancy and history of miscarriage. Occupational risk factors for thyroid cancer, with the exception of radioactive iodine, have not been well investigated. We conducted a case-cohort study nested in a cohort of 267,400 female textile workers in Shanghai, China, who had been followed for cancer incidence during 1989-1998. METHODS: The analysis included 130 incident thyroid cases and 3,187 subcohort non-cases. Reproductive history was determined by questionnaire at baseline. Historical exposures were reconstructed from work history and information on factory processes and exposures. Cox proportional hazards analysis was performed to estimate hazard ratios (HR) for reproductive factors and occupational exposures. RESULTS: Associations were observed between thyroid cancer and employment in jobs with 10 or more years of benzene exposure (HR 6.43, 95% CI: 1.08, 38) and formaldehyde exposure (HR 8.33, 95% CI: 1.16, 60). Administration workers also had an increased risk (HR 1.56, 95% CI: 1.08, 2.25). No associations between examined reproductive factors and thyroid cancer were observed in this study. CONCLUSIONS: Despite statistically imprecise risk estimates, the findings suggest potential associations with some occupational chemical exposures in this cohort of textile workers. PMID- 16220288 TI - Association of macrophage migration inhibitory factor gene -173 G/C polymorphism with prognosis in Turkish children with juvenile rheumatoid arthritis. AB - The objectives of this study were to determine genotypic and allelic frequencies of macrophage migration inhibitory factor (MIF) gene -173 G/C polymorphism in patients with juvenile rheumatoid arthritis (JRA) and to evaluate the association of the MIF -173 C allele with the outcome of JRA. Genomic DNA was collected from 67 JRA patients and 153 healthy individuals. To evaluate the association of the MIF -173 polymorphism with the outcome, we analyzed the data concerning the treatment regimen, duration of glucocorticoid treatment, score on the childhood health assessment questionnaire (C-HAQ) and the number of joints with active arthritis. Nonsignificant differences were observed between the study and control groups in the distribution of genotype and allele frequencies of the MIF gene 173 G/C polymorphism. In JRA patients, carrying a MIF -173 C allele, the number of disease modifying antirheumatic drugs required for the treatment was more, the duration of glucocorticoid treatment was significantly longer, and at the last visits the C-HAQ scores and the number of joints with active arthritis were significantly higher. MIF gene -173 C allele frequency did not differ between the controls and JRA patients. MIF -173 C allele did not confer increased susceptibility to JRA in our study group. Carriage of the MIF -173 C allele was found to be a strong predictor of poor outcome in all types of JRA. PMID- 16220290 TI - Kynurenic acid, an endogenous constituent of rheumatoid arthritis synovial fluid, inhibits proliferation of synoviocytes in vitro. AB - Kynurenic acid is an antagonist of ionotropic glutamate receptors. It has been found that glutamate antagonists inhibit proliferation of different human tumor cells. Since the hyperplasia of synovial fibroblasts is one of the most striking features of inflammatory arthritis, the main goals of this study were detection and quantification of kynurenic acid in synovial fluid obtained from patients with rheumatoid arthritis, and determination of its effect on proliferation of synoviocytes in vitro. Presence of kynurenic acid was determined by HPLC in all 58 samples of synovial fluid. The mean concentration was 15.89 pmol/ml. Kynurenic acid inhibited synoviocyte proliferation with the IC50 value of 5.9 mM. In subthreshold concentration of 0.3 mM it enhanced antiproliferative action of celecoxib and nimesulide. In conclusion, the presence of kynurenic acid in synovial fluid was documented in patients with rheumatoid arthritis. Its potential role as an endogenous substance, controlling synoviocyte proliferation can be suggested. PMID- 16220289 TI - Reactive arthritis following BCG immunotherapy for urinary bladder carcinoma: a systematic review. AB - Intravesical instillation of Bacillus Calmette-Guerin (BCG) is used with efficacy and safety in the treatment of patients with intermediate and high-risk superficial bladder carcinoma. Arthralgia and/or arthritis is one of the rare severe complications following intravesical BCG immunotherapy. We searched MEDLINE in order to analyze the frequency of this clinical complication, its pathogenesis and outcome. The electronic search was conducted using the following key words: "BCG immunotherapy" and "Arthritis, arthralgias and BCG immunotherapy". At the end of a process of abstract analysis, 48 papers were included in the systematic review. All the selected papers, except one that was a clinical review, described at least one case of arthritis after BCG therapy. The BCG immunotherapy resulted to be safe and efficacious in the treatment of bladder cancer; the development of reactive arthritis is rare and can evolve in a chronic process. The review of the literature highlighted that reactive arthritis following BCG intravesical instillation is a complication usually well controlled with the discontinuation of the immunotherapy and nonsteroidal anti-inflammatory drugs (NSAIDs) treatment. Only a small portion of patients with a particular genetic background will develop a chronic process. PMID- 16220291 TI - Novel transdermal photodynamic therapy using ATX-S10.Na(II) induces apoptosis of synovial fibroblasts and ameliorates collagen antibody-induced arthritis in mice. AB - We aimed to test the effect of transdermal photodynamic therapy (PDT) on synovial proliferation in vitro and in vivo, using a novel photosensitizer, ATX S10.Na(II). Synovial fibroblasts were obtained from patients with RA (RASF). Cell viability with or without PDT was determined by MTT assay. Cell morphology was examined by light and transmission electron microscopy. DNA fragmentation was labeled by TUNEL stain. Collagen antibody-induced arthritis (CAIA) was induced in DBA/1 mice, and the effects of transdermal PDT were evaluated by clinical and histological examination. PDT showed drug concentration-dependent and laser dose dependent cytotoxicity on RASF. TUNEL stain and TEM study revealed the induction of apoptotic cell death of RASF. Transdermal PDT significantly reduced clinical arthritis and synovial inflammation in this model of arthritis. These results suggest that transdermal PDT using ATX-S10.Na(II) might be a novel less invasive treatment strategy for small joint arthritis and tenosynovitis. PMID- 16220292 TI - Polymorphous adenocarcinoma of the breast. Report of three cases. AB - We report three cases of polymorphous adenocarcinoma (PLA) of the breast in 37-, 55- and 74-year-old women, respectively. The patients have no evidence of previous malignancy. The tumours consist of monotonous cells showing a wide spectrum of growth patterns: solid nests, trabeculae, tubules, cribriform structures, strands and fascicles reminiscent of polymorphous low-grade adenocarcinoma of salivary glands. To our knowledge, PLA has never been reported in the breast; therefore, this tumour should be added to the list of neoplastic lesions of the breasts that have the same features as those of the salivary glands. PMID- 16220293 TI - Historical, current and future perspectives on gastrointestinal and pancreatic endocrine tumors. AB - Gastrointestinal and pancreatic endocrine tumors are neoplasms of which the pathogenesis is not completely understood and of which the clinical behavior is difficult to predict. Originally, Masson suggested that the cell of origin was an endocrine cell derived from the gastrointestinal epithelium. However, Pearse showed that the endocrine cells throughout the body shared various features, among others the amine precursor uptake and decarboxylation (APUD) capacity, and postulated the neural crest as the common origin for all APUD cells, a hypothesis that received support from the scientific community for many years. Now, biologists start to elucidate the various transcription factors that drive gastrointestinal development, and it has become evident that Masson was presumably right. Transcription factors relevant for development may also operate during tumorigenesis, and their expression may determine tumor biology. With other genetic factors, they may play a role in the pathogenesis of gastrointestinal and pancreatic endocrine tumors, and perhaps, their expression will turn out to be of prognostic or therapeutic value. In this review, current knowledge on the development of endocrine cells, hypotheses on the origin of endocrine tumors, genetic alterations, and prognostic factors are discussed. It is suggested that the increasing understanding of the normal development of gastrointestinal and pancreatic endocrine cells, the accumulating data on genetic alterations in endocrine tumors and the reappraisal of the hypotheses on their pathogenesis formulated in the past may help in elucidating their pathogenesis and in more accurately predicting prognosis. PMID- 16220294 TI - Elevated hepatocyte paraffin 1 and neprilysin expression in hepatocellular carcinoma are correlated with longer survival. AB - Hepatocyte paraffin 1 (Hep Par 1) and neprilysin (CD10) are well-known markers of hepatocellular carcinoma (HCC). To assess their potential prognostic role, we conducted a retrospective analysis of 97 formalin-fixed and paraffin-embedded HCC from patients treated by surgery with curative intent, using standard immunohistochemical procedures and semiquantitative analysis. Strong Hep Par 1 expression and canalicular CD10 staining pattern were significantly correlated with smaller tumor size (p=0.007 and 0.04, respectively). On univariate analysis, longer overall survival was observed in patients with strong Hep Par 1 expression (p=0.0005) and in patients with a CD10can staining pattern (p=0.02). On multivariate analysis, the combined immunohistochemical score (CIS) obtained by addition of Hep Par 1 and CD10can scores and subtraction of cytoplasmic CD10 score was retained as the single most important prognostic factor (p=0.001). Patients with a CIS <4 had a 3.5-fold increased risk of death, as compared to those with a CIS >or=4. In conclusion, strong Hep Par 1 expression, presence of CD10can labeling, and absence of CD10cyt staining are favorable prognostic factors in HCC, which can be easily combined into a single immunohistochemical score for routine clinical use. PMID- 16220296 TI - Objective, planimetry-based assessment of megakaryocyte histological pictures in Philadelphia-chromosome-negative chronic myeloproliferative disorders: a perspective for a valuable adjunct diagnostic tool. AB - Philadelphia-chromosome-negative chronic myeloproliferative disorders (Ph- CMPDs) -essential thrombocythemia (ET), chronic idiopathic myelofibrosis (CIMF), and polycythemia vera (PV)--may show clinical and morphological similarities, particularly at the early stages. The differential diagnosis of Ph- CMPDs is important due to their different treatment and prognosis. Cytological features of megakaryocytes are considered valuable in this differentiation. To establish an objective measure of megakaryocyte dysplasia in Ph- CMPDs, we performed computer assisted morphometry of more than 4,000 cells from 20 cases of ET, 10 of CIMF, 10 of PV, and 10 controls. Megakaryocyte sets from three Ph- CMPDs differed significantly in respect to many planimetric parameters, but not a single shape or size parameter could have been used as a discriminative tool between the entities. However, the discriminant function analysis with the simultaneous assessment of 12 planimetric variables allowed for a proper classification of 20 of 20 ET, 10 of 10 PV, and 9 of 10 CIMF cases based solely on the morphometric features of megakaryocytes. Additionally, we identified certain new patterns of megakaryocytes specific for ET, PV, and CIMF, which, although not dominating in one Ph- CMPD, are unlikely to occur in two others. Objective measurements of megakaryocyte sizes and shapes may assist the diagnosis of Ph- CMPDs. PMID- 16220295 TI - Analysis of chromosomal imbalances in an elderly woman with a giant cell tumour. AB - Giant cell tumour (GCT) remains one of the most obscure and intensely studied bone tumours. In an effort to resolve questions regarding the genesis and clinical outcome of GCT, advances have been made recently in the identification of chromosomal abnormalities implicated in the tumour. Fusion of telomeres is very frequent in GCT, and this process may be associated with chromosome instability and tumour development. However, little emphasis has been placed on chromosomal imbalances in the molecular characterization of this disease. Here, we report the case of an 83-year-old woman diagnosed with GCT where local recurrence was observed after 11 months of the resection. Cytogenetic studies of the GCT showed a modal number of 46 chromosomes with telomeric associations on 11p and dicentric chromosomes. Moreover, clonal abnormalities, such as del(17p) and losses of chromosomes 4, 13 and 18 and gains on chromosome 7, were also detected. Interestingly, comparative genomic hybridisation (CGH) analysis revealed chromosomal imbalances with gains on chromosomes 1p31-q44, 6q12-q23 and 12q15-q22. Thus, the use of CGH expanded the information obtained by conventional cytogenetics and demonstrated that chromosomal imbalances were associated with the recurrence of the GCT. PMID- 16220297 TI - Expression of bFGF/FGFR-1 and vascular proliferation related to clinicopathologic features and tumor progress in localized prostate cancer. AB - Microvessel density (MVD) has been associated with progression of prostate cancer. Although basic fibroblast growth factor (bFGF) is a known endothelial mitogen, the prognostic role of bFGF and its receptor FGFR-1 in prostate cancer has been controversial. The aim of our study was to examine the tissue distribution and prognostic significance of bFGF, FGFR-1, and microvascular proliferation. Sections from 104 radical prostatectomy specimens were examined by factor VIII/Ki-67 staining for proliferating capillary index (PCI) and MVD, and tissue microarray sections were immunostained for bFGF and FGFR-1. Increased PCI (median 0.49%) was related to strong stromal expression of bFGF (P=0.003) but was without prognostic impact. Strong bFGF staining was associated with well differentiated tumors, no capsular penetration, low serum-prostate-specific antigen (s-PSA), low tumor cell proliferation, and increased time to biochemical failure (P=0.007), and was of independent prognostic importance in multivariate survival analysis. bFGF expression in vessels was associated with low MVD (P=0.0003). In contrast, strong tumor cell FGFR-1 expression was related to high preoperative s-PSA. Thus, increased stromal and vessel bFGF was associated with less aggressive tumors. Our findings indicate a complex relationship between bFGF/FGFR-1 expression and prognosis of prostate cancer. Vascular proliferation revealed no prognostic impact in this study. PMID- 16220298 TI - Osteoclast-rich tumor of the gastrointestinal tract with features resembling those of clear cell sarcoma of soft parts. AB - Clear cell sarcoma is a high-grade sarcoma with morphological features resembling those of malignant melanoma. An osteoclast-rich tumor of the gastrointestinal tract with features resembling those of clear cell sarcomas of soft parts is very rare. Herein, we report an unusual stomach tumor with microscopic and immunohistochemical characteristics of an osteoclast-rich tumor of the gastrointestinal tract with features resembling those of clear cell sarcomas of soft parts. The tumor cells were predominantly oval, admixed with some round and spindle elements arranged in nests and fascicles, and admixed with scattered osteoclast-like multinucleated giant cells. Neoplastic cells were positive for S 100 protein, and osteoclast-like multinucleated giant cells were immunoreactive to CD68. The unusual morphology of the tumor caused significant diagnostic difficulties. The differential diagnosis included gastrointestinal stromal tumor, primary or metastatic melanoma, and epithelioid malignant peripheral nerve sheath tumor. To the best of our knowledge, this is possibly the second description of an osteoclast-rich tumor of the gastrointestinal tract with features resembling those of clear cell sarcomas of soft parts. PMID- 16220299 TI - Claudins 1, 3, 4 and 5 in gastric carcinoma, loss of claudin expression associates with the diffuse subtype. AB - In this study expression of claudins 1, 3, 4 and 5 were studied in 118 cases of gastric carcinoma and compared with proliferation, apoptosis and E-cadherin expression. Expression of all these claudins could be seen in gastric carcinoma, most prominently for claudin 4, and least expression was found for claudin 5. All claudins showed significantly more expression in gastric carcinomas of intestinal type. Their expression was significantly associated with each other. Expression of claudins 4 and 5 was associated with E-cadherin. Strong expression of claudin 5 was associated with higher cell proliferation and apoptosis. Claudin 3 expression had an association with a better prognosis of the patients, especially in the intestinal type. The results show that expression of claudins 1, 3, 4 and 5 is lower in diffuse-type gastric carcinomas. Possibly they play a role in determining the diffuse phenotype and loose cohesion of cells in diffuse type of gastric carcinoma in a similar manner as E-cadherin. The loss of their expression does not clearly associate with poorer prognosis of the patients, except for claudin 3, where strong expression was associated with a better outcome of the patients, a feature especially related to intestinal-type tumours. PMID- 16220301 TI - Unilateral (probable) moyamoya disease: long-term follow-up of seven cases. AB - OBJECTS: Some cases of unilateral involvement have angiographic findings on the affected side which resemble those of definite cases of moyamoya disease (MMD). We analyzed these cases to determine whether the entity "unilateral MMD" actually exists and whether unilateral involvement represents only the early phase of the pathologic process found in definitive bilateral disease. METHODS: Between 1988 and 2000, seven cases of unilateral MMD were evaluated. The male-to-female ratio was 2:5, mean age was 5.1 years (7 months to 8 years), and mean follow-up duration was 64.7 months. The presenting symptoms of these cases were similar to those of definite MMD, and no familial occurrence was found. Two of the seven unilateral MMD cases showed progression to bilateral involvement. However, the remaining cases showed no evidence of contralateral progression during the follow up period. CONCLUSIONS: Most cases of unilateral MMD seem to be an unusual form of stenoocclusive process of the proximal intracranial arteries, distinguishing the condition from definite MMD. Surgical treatment on the symptomatic side and close observation for signs of bilateral involvement are necessary. PMID- 16220300 TI - [Implications of estrogens and their receptors for the development and progression of prostate cancer]. AB - The recent discovery of the estrogen receptors alpha and beta (ERalpha, ERbeta) and the progesterone receptor (PR) in human prostate tissue offers new insights into the role of estrogens and their receptors in prostate cancer development and tumor progression. The differentiation compartment of the prostatic epithelium (secretory luminal cells) expresses high levels of ERbeta, while the ERalpha is restricted to the proliferation compartment (basal cells). In high grade prostatic intraepithelial neoplasia (HGPIN), ERalpha gene expression extends to luminal cells and thus may mediate cancerogenic effects of estrogens on the dysplastic epithelium. Conversely, the ERbeta is downregulated in HGPIN indicating that the chemopreventive effects of phytoestrogens mediated by the ERbeta are partially lost. Irrespective of grades and stages, prostate cancer retains high levels of the ERbeta which is partially lost in androgen-insensitive stages of the disease. In contrast with breast cancer, the presence of the ERalpha and the PR is a late event in prostate cancer progression. At least 30% of metastatic and androgen-insensitive tumors express high levels of the PR indicating that these tumors harbor a functional ERalpha. The antiestrogen Raloxifene has growth-inhibitory effects on androgen-insensitive prostate cancer cells in vitro and induces the apoptotic cell death in a dose-dependent fashion. These data provide a rational for clinical trials to study the efficiency of antiestrogens in the medical treatment of advanced prostate cancer. PMID- 16220302 TI - The situation of nuclear medicine in Central and Eastern Europe. AB - This paper reviews the situation of nuclear medicine in Central and Eastern Europe (CEE), i.e. a group of 15 countries lying east of the Oder river-Trieste line and west of the present/planned EU eastern border. Together these countries have 106.6 million inhabitants, i.e. about a quarter of the population of the "European economic space". Its nuclear medicine, however, represents less than 10% of European nuclear medicine manpower and equipment. In these countries there are at least 245 nuclear medicine departments with 661 nuclear medicine specialists and at least 376 gamma cameras. There are six dedicated PET units and three PET/SPECT centres, as well as one manufacturer of gamma cameras, six radiopharmaceutical manufacturers and two nuclear medicine scientific journals. The biggest nuclear medicine communities are in the Czech Republic, Hungary and Poland. The scientific input of the CEE countries to European science is moderate about 10% when measured by EANM congress abstracts, and 5% when measured by the number of papers in MEDLINE-indexed journals. Mean European bibliometric parameters are approached only by Hungary and-to some extent-the Czech Republic. This survey indicates the need for international cooperation to improve the level of nuclear medicine in the CEE countries so that it reaches European standards. The potential of these countries could also be better used to the benefit of European nuclear medicine. PMID- 16220303 TI - Lymphoscintigraphic sentinel node identification in patients with breast cancer: the role of SPECT-CT. AB - PURPOSE: Lymph node status is a major factor in determining the stage, appropriate therapy and outcome in patients with breast cancer. It is therefore of clinical importance to accurately identify all sentinel nodes (SNs) for each individual tumour before surgery. The purpose of this study was to assess the role of SPECT-CT lymphoscintigraphy in SN identification in patients with breast cancer. METHODS: Lymphoscintigraphy comprising planar and SPECT-CT acquisition was performed in 157 consecutive patients with breast cancer (mean age 54.7+/ 10.6, range 27-81 years) with a palpable mass (n=100), with a non-palpable mass (n=52) or post lumpectomy (n=5). Planar and SPECT-CT images were interpreted separately and the two imaging techniques were compared with respect to their ability to identify hot nodes. RESULTS: Planar imaging alone was negative for identification of hot nodes in 15% of the patients. SPECT-CT alone was negative in 10% and both techniques were negative in 9% of the patients. Forty-six of the total of 361 (13%) hot nodes identified by lymphoscintigraphy were detected only on SPECT-CT, including 21 nodes obscured by the scattered radiation from the injection site, nine adjacent nodes misinterpreted on planar images as a single node and 16 nodes which were missed on planar images and detected on SPECT data. SPECT-CT detected additional sites of drainage unexpected on planar images, including axillary (n=23 patients), internal mammary (n=5 patients), interpectoral (n=3 patients) and intramammary (n=2 patients) lymph node sites. Fourteen of the 329 (4%) hot lesions seen on planar images were false positive non-nodal sites of uptake that were accurately assessed by SPECT-CT and further validated by surgery. In a single patient, SPECT-CT was negative while planar images identified the SN. CONCLUSION: SPECT-CT may improve the preoperative localisation of draining nodes in patients with breast cancer. It may detect hot nodes missed by planar imaging, exclude non-nodal false positive sites of uptake and accurately localise axillary and extra-axillary nodes. PMID- 16220304 TI - FDG-PET in immunocompetent patients with primary central nervous system lymphoma: correlation with MRI and clinical follow-up. AB - PURPOSE: The role of FDG-PET in primary central nervous system lymphoma (PCNSL) is unclear. It was the aim of this study to investigate the role of FDG-PET in detecting PCNSL and in predicting response to chemotherapy. METHODS: An FDG-PET scan of the brain was performed in 15 patients with histologically proven PCNSL (16 PET examinations, Siemens ECAT EXACT). PET was planned to investigate patients at the time of primary diagnosis, after chemotherapy and at the time of suspected relapse in seven, five and three cases, respectively. All except two patients simultaneously underwent MRI of the brain. FDG-PET results were correlated with histological results after stereotactic biopsy (primary diagnosis group) and with clinical data and MRI during follow-up. RESULTS: Six of the seven patients in the primary diagnosis group demonstrated a true positive finding (86%). In one of the true positive PET patients, there were two tumour lesions, one of which was only detectable on the FLAIR MRI sequence. In five patients, FDG PET showed no sign of PCNSL during ongoing chemotherapy. These results were confirmed by the clinical follow-up (mean 26.6 months). MRI demonstrated minimal residual disease which had disappeared on further follow-up MRI in three of these five patients at the time of PET scanning. Recurrence of disease was confirmed concordantly by FDG-PET and MRI in three different patients. The standardised uptake value of all tumours was 10.2 (4.3-13.7). CONCLUSION: PCNSLs demonstrate high FDG uptake and can be diagnosed by FDG-PET with high sensitivity. It seems that FDG-PET is suitable for early therapeutic monitoring after chemotherapy. PMID- 16220305 TI - 99mTc-interleukin-2 scintigraphy for the in vivo imaging of vulnerable atherosclerotic plaques. AB - PURPOSE: Several histopathological studies have demonstrated that vulnerable plaques are enriched in inflammatory cells. The aims of this study were: (1a) to test the ability of 99mTc-labelled interleukin-2 (99mTc-IL2) to bind to IL2R positive (IL2R+) cells in carotid plaques and (1b) to correlate the plaque uptake of 99mTc-IL2, measured in vivo, with the number of IL2R+ cells within the plaque, measured ex vivo by histology (transversal study, TS), and (2) to evaluate changes in 99mTc-IL2 uptake in plaques, before and after treatment with a statin or a hypocholesterolaemic diet (longitudinal study, LS). METHODS: Ultrasound scan was performed for plaque characterisation and localisation. Fourteen patients (16 plaques) eligible for endoarterectomy were recruited for the TS and underwent 99mTc-IL2 scintigraphy before surgery. Nine patients (13 plaques) were recruited for the LS; these patients received atorvastatin or a standard hypocholesterolaemic diet and 99mTc-IL2 scintigraphy was performed before and after 3 months of treatment. RESULTS: The degree of 99mTc-IL2 uptake was expressed as the plaque/background (T/B) ratio. In patients from TS, T/B ratios correlated with the percentage of IL2R+ cells at histology (r = 0.707; p = 0.002) and the number of IL2R+ cells at flow cytometry (r = 0.711; p = 0.006). No correlations were observed between ultrasound scores and either scintigraphic or histological findings. In patients from the LS, the mean 99mTc-IL2 uptake decreased in statin-treated patients (1.75+/-0.50 vs 2.16+/-0.44; p = 0.012), while it was unchanged in the patients on the hypocholesterolaemic diet (2.33+/ 0.45 vs 2.34+/-0.5). CONCLUSION: 99mTc-IL2 accumulates in vulnerable carotid plaques; this accumulation is correlated with the amount of IL2R+ cells and is influenced by lipid-lowering treatment with a statin. PMID- 16220306 TI - Correlation of FDG-PET findings with histopathology in the assessment of response to induction chemoradiotherapy in non-small cell lung cancer. AB - PURPOSE: The objective of this study was to evaluate the ability of FDG-PET to predict the response of primary tumour and nodal disease to preoperative induction chemoradiotherapy in patients with non-small cell lung cancer (NSCLC). METHODS: FDG-PET studies were performed before and after completion of chemoradiotherapy prior to surgery in 26 patients with NSCLC. FDG-PET imaging was performed at 1 h (early) and 2 h (delayed) after injection. Semi-quantitative analysis was performed using the standardised uptake value (SUV) at the primary tumour. Percent change was calculated according to the following equation: [see text]. Based on histopathological analysis of the specimens obtained at surgery, patients were classified as pathological responders or pathological non responders. The clinical nodal stage on the post-chemoradiotherapy PET scan was visually determined and compared with the final pathological stage. RESULTS: Eighteen patients were found to be pathological responders and eight to be pathological non-responders. SUV(after) values from both early and delayed images in pathological responders were significantly lower than those in pathological non-responders. The percent change values from early and delayed images in the pathological responders were significantly higher than those in the pathological non-responders. The post-chemoradiotherapy PET scan accurately predicted nodal stage in 22 of 26 patients. CONCLUSION: FDG-PET may have the potential to predict response to induction chemoradiotherapy in patients with NSCLC. PMID- 16220307 TI - The role of 18F-fluorodeoxyglucose positron emission tomography in gestational trophoblastic tumours: a pilot study. AB - PURPOSE: We conducted a pilot trial to evaluate the value of 18F fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) in gestational trophoblastic tumours (GTTs). METHODS: Patients with placental site trophoblastic tumour (PSTT), high-risk GTT (World Health Organisation score > or =8, disease onset at postpartum or greater than 6 months after antecedent pregnancy), metastatic GTT, recurrent/resistant GTT after chemotherapy, or post-molar GTT with unexplained abnormal beta-hCG regression and patients undergoing re evaluation after salvage treatment were enrolled. PET was undertaken within 1 week after computed tomography (CT). Clinical impacts of additional PET were determined on a scan basis. RESULTS: A total of 14 patients were recruited. Sixteen PET scans were performed, with one patient having three serial studies. Benefits of additional PET were seen in 7 of 16 (43.8%) scans; these benefits included disclosure of chemotherapy-resistant lesions (n = 2), exclusion of false positive CT lesions (n = 1), detection of an additional lesion not found by conventional imaging (n = 1) in high-risk GTT at the start of primary chemotherapy, and confirmation of complete response to treatment for PSTT or to salvage therapy for recurrent/resistant GTT (n = 3). On the other hand, in two instances there were false-negative PET findings, six scans yielded no benefit, and one showed an indeterminate lesion. CONCLUSION: Our preliminary results suggest that 18F-FDG PET is potentially useful in selected patients with GTT by providing precise mapping of metastases and tumour extent upfront, by monitoring treatment response and by localising viable tumours after chemotherapy. A larger study is necessary to further define the role of 18F-FDG PET in GTT. PMID- 16220308 TI - Human T-lymphotropic virus type-1 (HTLV-1) in Israeli patients and their family relatives and its transmission to rats. AB - We tested the possibility that lymphocytes, sera and saliva, obtained directly from healthy human T-lymphotropic virus type 1 (HTLV-1) carriers and patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) of Iranian Mashhadi origin, as well as lymphocytes from patients with mycosis fungoides (MF) and their family relatives (MFR), may be infective. Peripheral blood mononuclear cells (PBMC), sera, PBMC cultured with phytohaemagglutinin A and phorbol myristate acetate, cell-free supernatant from these cultures, saliva cells and cell-free saliva were injected into adult WKA (n=107) and F344 (n=47) female rats. The appearance of anti-HTLV-1 antibodies in the rat sera was tested by particle agglutination assay and ELISA, and positive results were confirmed by western blot assay. Higher titers (1:1024) of anti-HTLV-1 antibodies were found in the F344 rats as compared to the WKA rats (1:256). The PA agglutination test was the most sensitive for the detection of HTLV-1 antibody. The HTLV-1 provirus was detected in both strains of rats infected with body fluids and cells from the Iranian Mashhadi Jews, in various organs (PBMC, spleen, thymus, salivary glands, spinal cord, kidney and brain) by nested PCR. However, the HTLV-1 provirus was not detected in 100% of the rats. The negative rats were only immunized and not infected. The spleen, thymus, spinal cord and salivary glands of the seropositive rats were found to be infectious and to transmit the HTLV-1 to healthy rats. F344 rats infected with PBMC cultures obtained from HTLV-1 antibody positive MF patients and their MFR who were only 20% positive showed anti-HTLV-1 antibodies, but only in 20% of rats without showing the HTLV-1 provirus; these rats were probably not infected but only immunized. This is one of the few studies on the transmission of HTLV-1 to rats by inoculation with human infectious fluids or cells from HTLV-1 infected healthy carriers (42%), HAM/TSP patients of Iranian Mashhadi origin (58%) as well as lymphocyte cultures obtained from HTLV-1 antibody positive MF and MFR of nonIranian origin. PMID- 16220309 TI - Cellular receptor for Pixuna virus in chicken embryonic fibroblasts. AB - In this study, we describe the isolation and partial characterization of a Pixuna virus receptor, which is a component of a plasma membrane fraction of chicken embryo fibroblast (CEF). Polyclonal antiserum was prepared from rabbits immunized with the membrane fraction. Said polyclonal antiserum reacted in a similar way as monoclonal antibodies raised against the membrane fraction. Both antisera were able to prevent CEF and Vero cells from infection with Pixuna virus. Immunofluorescence studies suggested that the receptors found in the fibroblasts and in the Vero cells shared at least some epitopes. The Western blot analysis of the purified membrane fraction antigens, which reacted with the monoclonal and polyclonal antibodies, detected a double band with a molecular mass of approximately 60 kDa. Not only immunofluorescence staining but also electron and immunoelectron microscopy studies evidenced the receptor localization in the plasma membrane. In this manner, we reported the isolation and partial characterization of a new Pixuna virus receptor in the plasma membrane of chicken embryo fibroblasts in culture. The data obtained demonstrated the receptor significance for the penetration of Pixuna virus into fibroblasts and mammalian cell and the related importance of designing new antiviral drugs by blocking the mechanism of receptor penetration of the virus into the cells. PMID- 16220310 TI - Ethical issues in the daily medical care of children. AB - The main ethical imperative of all paediatric actions is the demand to do everything "in the best interests of children". Relevant guidelines can be derived from the UN Declaration on the Rights of Children, whereupon every child has the fundamental right to life and dignity, and is entitled to optimal medical care. Paediatric care in general includes the responsibility to achieve the highest level of knowledge, consideration of the child-specific somatic, mental and social development, empathic and trustworthy communication with the child and parents, observance of the recommendations of the Charter of the Rights of Children in Hospital, and cooperation with experts in related professions. Good communication is based on respect for the dignity of the child as a person and on the use of child-specific language, recognizing the rights of the child to be involved in consent or assent. Good clinical practice dictates and demands high standards of practice in therapeutics, research and medical interventions involving children. Decision making in extreme situations with regard to continuation, withholding or withdrawing life supporting measures is amongst the most complex and ethically difficult tasks of a doctor. Ethical issues with regard to neglect, maltreatment, abuse and addictions involving children need scrupulous consideration. Paediatricians have a prime responsibility to promote and protect the well being of children. PMID- 16220311 TI - Decitabine: a historical review of the development of an epigenetic drug. AB - The development of decitabine from its synthesis in 1964 to the submission of a registration file has been described. Although the unique DNA-demethylating capacity of decitabine is known for a long time, its application is under continuing investigation. The use of decitabine in MDS, AML, CML, stem cell transplant, sickle cell anemia and thalassemia looks promising. The epigenetic dose seems lower than the cytotoxic dose. Whereas most drugs have matured after 40 years, decitabine is only at the beginning of a new development phase in epigenesis. PMID- 16220312 TI - Successful treatment of disseminated fusariosis with high dose liposomal amphotericin-B in a patient with acute lymphoblastic leukemia. PMID- 16220313 TI - Anatomy of the lateral collateral ligament: a cadaver and histological study. AB - This paper describes the anatomy of the lateral collateral ligament (LCL). The dimensions of the ligament and its femoral and fibular attachments are given. The relationships between the LCL and other anatomical structures are described, particularly the terminal fiber branches of the biceps femoris. The histological features of the ligament fibers and their osseous attachments are also described. PMID- 16220314 TI - [Intraocular lenses for the correction of refraction errors. Part II. Phakic posterior chamber lenses and refractive lens exchange with posterior chamber lens implantation]. AB - In this overview, the current status of intraocular lens surgery to correct refractive error is reviewed. The interventions are divided into additive surgery with intraocular lens implantation without extraction of the crystalline lens (phakic intraocular lens, PIOL) or removal of the crystalline lens with implantation of an IOL (refractive lens exchange, RLE). Phakic IOLs are constructed as angle-supported or iris-fixated anterior chamber lenses and posterior chamber lenses which are fixated in the ciliary sulcus. The implantation of phakic IOLs has been demonstrated to be an effective, safe, predictable and stable procedure to correct higher refractive errors. Complications are rare and differ for the three types of PIOL; for posterior chamber lenses these are mainly cataract formation and pigment dispersion. RLE is preferable in cases of high ametropia in which the natural lens has lost its accommodative effect. The main complications for myopic RLA include retinal detachment, while hyperopic refractive lens exchange may be associated with surgical problems in the narrower anterior eye segment. PMID- 16220316 TI - [Hemangiopericytoma (extrapleural solitary fibrous tumour). Exemplified and discussed on the basis of two cases]. AB - BACKGROUND: Hemangiopericytoma was first mentioned in 1942. It is a very uncommon potentially malignant vascular tumour which can occur at every site of the body. According to the WHO, most of the tumours formerly diagnosed as hemangiopericytomas are considered to be extrapleural solitary fibrous tumours. The diagnosis of "hemangiopericytoma" is now only determined if a constant histological picture of hemangiopericytoma is present. The tumour can lead to lymphogenous or hematogenous metastasis. The major location of occurrence is the cutis and subcutis. It originates from the pericytes of the vascular wall. This is reflected in its vascular character and therefore the hemangiopericytoma might clinically be mistaken for a hemangioma. CASE REPORTS: Patient 1: 60 years, female; diagnosis: malignant suboccipital hemangiopericytoma; size: 4.9 x 4.5 x 4.2 cm; pT1bNXMX L0 V0 Pn0; stage IA; grading G1; R0.Patient 2: 38 years, male; diagnosis: benign hemangiopericytoma infraorbital left; size 1.5 x 1.5 x 1.5 cm. DISCUSSION: Most often the hemangiopericytoma becomes clinically conspicuous as a slowly growing, painless swelling. The consistency ranges from soft to dense, and the color is greyish-blue. The slow and painless growth carries the danger of a clinically wrong diagnosis and thus delayed therapy. The histological diagnosis of hemangiopericytoma is determined by biopsy. Besides histology, MRI and angiography are methods that can be employed to diagnose hemangiopericytoma. The therapy of choice is the complete tumour-resection with a safety margin of 1 cm. In the case of an aggressive growth pattern, adjuvant postoperative radiotherapy is recommended. Until now there has been no documented specific therapy concept for managing incomplete resection and the occurrence of metastasis. There are reports about chemo- and radiotherapy either on their own or combined which evidence differing degrees of success. Lifelong monitoring is necessary because recrudescences and metastases can occur even decades later. PMID- 16220315 TI - Outcome and changes over time in survival following severe burns from 1985 to 2004. AB - OBJECTIVE: To investigate outcome in severely burned patients over a 20-year period and to evaluate survival over time. DESIGN AND SETTING: Historical cohort in a six-bed burn unit of a 1060-bed university hospital. PATIENTS: 1385 patients admitted to the burn unit over a 20-year period. MEASUREMENTS AND RESULTS: Outcome was evaluated in relation to the presence of three major risk factors for death: age 60 years or over, total burned surface area 40% or more, and the presence of inhalation injury. Overall mortality was 7.1%. When zero, one, two, or three risk factors were present, mortality was respectively 0.5%, 9.9%, 48.0%, and 90.5%. Over the study period the average proportional total burned surface area decreased as did mortality. The survival benefit was significant among patient groups with one or two risk factors present. Multivariate regression analysis adjusting for risk factors for death confirmed that survival improved over time (odds ratio 0.73 per 5-year period). CONCLUSIONS: Global mortality following burns is low, and nearly all patients who die had at least one risk factor present. In the presence of three risk factors the prognosis following burns is particularly compromised. Taking into account that our patients over the past 20 years have been progressively less extensively burned and hence have a lesser at risk for death, survival following severe burns has continued to improve. PMID- 16220317 TI - Quantification of benzodiazepines in whole blood and serum. AB - A high-performance liquid chromatography method for the determination of benzodiazepines and their metabolites in whole blood and serum using mass spectrometry (MS) and photodiode array (PDA) detection is presented. The combination of both detection types can complement each other and provides extensive case relevant data. The limits of quantification (LOQ) with the MS detection lie between 2 and 3 microg/l for the following benzodiazepines or metabolites: 7-amino-flunitrazepam, alprazolam, desalkyl-flurazepam, desmethyl flunitrazepam, diazepam, flunitrazepam, flurazepam, alpha-hydroxy-midazolam, lorazepam, midazolam, nitrazepam, nordazepam and oxazepam, respectively 5 microg/l for lormetazepam and 6 microg/l for bromazepam. The LOQ of clobazam determined with the PDA detector is 10 microg/l. A convenient approach for determining the measurement uncertainty of the presented method--applicable also for other methods in an accreditation process--is presented. At low concentrations (<10 microg/l), measurement uncertainty was estimated to be about 50%, and at concentrations >180 microg/l, it was estimated to be about 15%. One hundred and twenty-eight case data acquired over 1 year are summarised. PMID- 16220318 TI - Does imaging technology overcome problems of conventional postmortem examination? A trial of computed tomography imaging for postmortem examination. AB - We used a mobile computed tomography (CT) unit for postmortem examinations of deceased subjects to see how many mistakes on cause-of-death diagnoses were made in Japan. In 5 of 20 cases, the cause of death determined by CT was different from the diagnosis made by superficial postmortem examination. In one case, the superficial examination suggested no trauma, whereas a subdural hematoma was found on cranial CT images. We concluded that postmortem examinations in Japan were not effective when screening for crimes or accidents. Using a mobile CT scanner in postmortem examination may be a viable method of screening for causes of deaths, although it cannot be used as a substitute for autopsy. PMID- 16220324 TI - Cytomodulation of interleukin-2 effect by L-2-oxothiazolidine-4-carboxylate on human malignant melanoma. AB - Glutathione (GSH), the most prevalent intracellular non-protein thiol, plays an important role in the interleukin-2 (IL-2)-induced proliferative activity of normal and tumour cells expressing IL-2 receptor (IL-2R). In the present study, we investigate the effect of IL-2 on proliferation of the human melanoma A375 cell line, and the possible selective cytomodulation effect of this cytokine by L 2-oxothiazolidine-4-carboxylate (OTZ) on these melanoma cells and on human peripheral blood mononuclear cells (PBMCs). We found that recombinant IL-2 (rIL 2) significantly increased the proliferation rate of A375 melanoma cells, which was associated with an increase in GSH levels, the enhancement of IL-2Ralpha expression and the endogenous production of IL-2 in these tumour cells. In contrast, OTZ decreased GSH content and the proliferation rate of A375 cells, and abrogated the growth-promoting effects of rIL-2. Thus, compared to cells treated with rIL-2, pre-treatment with OTZ reduced IL-2Ralpha expression, and also decreased the consumption of rIL-2 and the endogenous secretion of IL-2 by these tumour cells. With regard to PBMCs, the combination of OTZ plus rIL-2 resulted in a more rapid and greater increase of IL-2Ralpha expression than rIL-2 alone, with the proliferation rate being similar in the first 24 h, but with a lower PBMC' count found thereafter compared to rIL-2 treatment alone. These results suggest that OTZ plays a crucial role in obtaining a selective cytomodulation of rIL-2, enabling it to exert its growth-promoting effect on normal cells, but not on melanoma cells, thereby possibly improving biochemotherapy with rIL-2. PMID- 16220325 TI - Defining MHC class II T helper epitopes for WT1 tumor antigen. AB - The product of Wilms' tumor gene 1 (WT1) is overexpressed in diverse human tumors, including leukemia, lung and breast cancer, and is often recognized by antibodies in the sera of patients with leukemia. Since WT1 encodes MHC class I restricted peptides recognized by cytotoxic T lymphocytes (CTL), WT1 has been considered as a promising tumor-associated antigen (TAA) for developing anticancer immunotherapy. In order to carry out an effective peptide-based cancer immunotherapy, MHC class II-restricted epitope peptides that elicit anti-tumor CD4(+) helper T lymphocytes (HTL) will be needed. In this study, we analyzed HTL responses against WT1 antigen using HTL lines elicited by in vitro immunization of human lymphocytes with synthetic peptides predicted to serve as HTL epitopes derived from the sequence of WT1. Two peptides, WT1(124-138) and WT1(247-261), were shown to induce peptide-specific HTL, which were restricted by frequently expressed HLA class II alleles. Here, we also demonstrate that both peptides reactive HTL lines were capable of recognizing naturally processed antigens presented by dendritic cells pulsed with tumor lysates or directly by WT1+ tumor cells that express MHC class II molecules. Interestingly, the two WT1 HTL epitopes described here are closely situated to known MHC class I-restricted CTL epitopes, raising the possibility of stimulating CTL and HTL responses using a relatively small synthetic peptide vaccine. Because HTL responses to TAA are known to be important for promoting long-lasting anti-tumor CTL responses, the newly described WT1 T-helper epitopes could provide a useful tool for designing powerful vaccines against WT1-expressing tumors. PMID- 16220326 TI - Tumor stroma-associated antigens for anti-cancer immunotherapy. AB - Immunotherapy has been widely investigated for its potential use in cancer therapy and it becomes more and more apparent that the selection of target antigens is essential for its efficacy. Indeed, limited clinical efficacy is partly due to immune evasion mechanisms of neoplastic cells, e.g. downregulation of expression or presentation of the respective antigens. Consequently, antigens contributing to tumor cell survival seem to be more suitable therapeutic targets. However, even such antigens may be subject to immune evasion due to impaired processing and cell surface expression. Since development and progression of tumors is not only dependent on cancer cells themselves but also on the active contribution of the stromal cells, e.g. by secreting growth supporting factors, enzymes degrading the extracellular matrix or angiogenic factors, the tumor stroma may also serve as a target for immune intervention. To this end several antigens have been identified which are induced or upregulated on the tumor stroma. Tumor stroma-associated antigens are characterized by an otherwise restricted expression pattern, particularly with respect to differentiated tissues, and they have been successfully targeted by passive and active immunotherapy in preclinical models. Moreover, some of these strategies have already been translated into clinical trials. PMID- 16220327 TI - Assessment of methylphenidate-induced changes in binding of continuously infused [(11)C]-raclopride in healthy human subjects: correlation with subjective effects. AB - RATIONALE: The dopaminergic system has been implicated in the pathogenesis and treatment of a variety of neuropsychiatric disorders. It has been shown that information on endogenous dopamine (DA) release can be obtained noninvasively by combining positron emission tomography with a dopaminergic challenge. This approach is based on the assumption that an injected radiolabeled ligand competes with the neurotransmitter for the same receptor. Increases in DA release will therefore result in a decreased binding of the radioligand. OBJECTIVES: We investigated the effect of the DA reuptake blocker methylphenidate (MP) on the binding of the D(2) receptor ligand [(11)C]-raclopride (RAC). METHODS: The effect of a 0.25 mg/kg intravenous dose of MP was studied in six healthy volunteers. RAC was administered as a bolus followed by constant infusion, and subjective effects were assessed using verbal rating scales. RESULTS: Control scans without MP administration showed that the mean RAC binding reached stable values approximately 30 min after start of the infusion. MP administration induced a 24% decrease in RAC binding in the total striatum. Correlations were found between the MP-induced change in euphoria and the percent change in binding potential (DeltaBP) in the dorsal striatum and between baseline anxiety and DeltaBP in the dorsal and middle striatum. We also found a negative correlation between baseline BP in the dorsal striatum and change in euphoria. CONCLUSIONS: Our results comply with previous findings, indicating the feasibility of the bolus infusion design combined with a relatively low MP dose to study dopaminergic (dys)function. PMID- 16220328 TI - 2-MPPA, a selective glutamate carboxypeptidase II inhibitor, attenuates morphine tolerance but not dependence in C57/Bl mice. AB - RATIONALE AND OBJECTIVES: We have recently reported that conditioned morphine reward and tolerance to its antinociceptive effect, but not expression of morphine dependence, were attenuated by 2-(phosphonomethyl)pentanedioic acid (2 PMPA), a prototypic inhibitor of glutamate carboxipeptidase II (GCP II), which is an enzyme responsible for the supply of glutamate. In the present study, we investigated in more detail the effects of GCP II inhibition on opioid dependence and tolerance to its antinociceptive effect in C57/Bl mice using a novel GCP II inhibitor. RESULTS: The treatment with 2-(3-mercaptopropyl)pentanedioic acid (2 MPPA; 60 but not 10 or 30 mg/kg) prevented the development of morphine tolerance without affecting acute morphine antinociception. 2-MPPA at 30 and 60 mg/kg did not prevent the development of dependence induced by 10 and 30 mg/kg of morphine. The study on opioid withdrawal syndrome, i.e., expression of opioid dependence, demonstrated that 2-MPPA potentiated jumping behavior and teeth chattering but attenuated chewing and ptosis. None of these opioid withdrawal signs were affected by 2-MPPA in morphine nondependent mice. Pretreatment with the mGluR II antagonist LY341495 (1 mg/kg) reversed the 2-MPPA-induced increase or decrease in opioid withdrawal signs in morphine-dependent mice. 2-MPPA (60 mg/kg) administered for 7 days with morphine did not affect brain concentration of this opiate. CONCLUSIONS: The present findings suggest complex effects of GCP II inhibition on morphine dependence and tolerance and imply a role of mGluR II in the actions of 2-MPPA. PMID- 16220329 TI - Enhanced effect of dopaminergic stimulation on prepulse inhibition in mice deficient in the alpha subunit of G(z). AB - RATIONALE: G(z) is a member of the G(i) G protein family associated with dopamine D2-like receptors; however, its functions remain relatively unknown. The aim of the present study was to investigate prepulse inhibition (PPI) of acoustic startle, locomotor hyperactivity and dopamine D2 receptor binding in mice deficient in the alpha subunit of G(z). METHODS: We used automated startle boxes to assess startle and PPI after treatment with saline, amphetamine, apomorphine or MK-801. We used photocell cages to quantitate locomotor activity after amphetamine treatment. Dopamine D2 receptor density was determined by autoradiography. RESULTS: Startle responses and baseline PPI were not different between the Galpha(z) knockout mice and wild-type controls (average PPI 46+/-4 vs 49+/-3%, respectively). Amphetamine treatment caused a marked disruption of PPI in Galpha(z) knockouts (average PPI 22+/-2%), but less so in controls (average PPI 42+/-3%). Similar genotype-dependent responses were seen after apomorphine treatment (average PPI 23+/-3% vs 40+/-3%), but not after MK-801 treatment (average PPI 29+/-5 vs 33+/-2%). Amphetamine-induced locomotor hyperactivity was greater in Galpha(z) knockouts than in controls. There was no difference in the density of dopamine D2 receptors in nucleus accumbens. CONCLUSIONS: Mice deficient in the alpha subunit of G(z) show enhanced sensitivity to the disruption of PPI and locomotor hyperactivity caused by dopaminergic stimulation. These results suggest a possible role for G(z) in neuropsychiatric illnesses with presumed dopaminergic hyperactivity, such as schizophrenia. PMID- 16220330 TI - Simultaneous intra-accumbens remifentanil and dopamine kinetics suggest that neither determines within-session operant responding. AB - RATIONALE: The ultra-short-acting mu opioid agonist analgesic/anesthetic remifentanil (RMF) is extremely rapidly eliminated from blood (half-life in rats, 0.3-0.7 min). This extremely fast elimination is thought to be the main reason why RMF maintains such high rates of responding in animal operant-conditioning models of drug addiction. OBJECTIVE: The present study investigated if such a fast elimination of RMF also occurs in the extracellular space of the brain, i.e., in the pharmacokinetic compartment that is thought to be ultimately mediating the reinforcing effect, and hence, the abuse liability of drugs. METHODS: Nucleus accumbens (NAC) RMF and dopamine (DA) were simultaneously quantified by in vivo microdialysis followed by tandem mass spectrometry both in rats that traversed an alley to receive intravenous injections of 0.032 mg kg(-1) RMF in an operant runway procedure (contingent RMF) and in rats that passively received RMF in the runway (noncontingent RMF). RESULTS: Regardless of the mode of administration (i.e., contingent or noncontingent), intra-accumbens RMF peaked in the first 10-min sample and decreased exponentially with a t(1/2) of 10.0+/ 1.2 min (N=31). RMF-stimulated DA peaked in the 10-min sample immediately after the RMF peak and decreased with a time course very similar to that of RMF. Crosscorrelation of the NAC RMF and NAC DA curves showed them to be tightly synchronized. Noncontingent single-dose RMF was eliminated from the whole brain with a half-life of 1.1+/-0.2 min and from blood with a half-life of 0.3 min or less. The comparison of blood-vs-brain RMF pharmacokinetics with rat RMF self administration behavior, either in operant runway (present study) or in lever press-based operant-conditioning procedures, suggests that titration of blood RMF, whole-brain RMF, intra-accumbens RMF, or accumbal DA levels (assessed with the limited temporal resolution of in vivo microdialysis) does not determine a rat's decision to reemit a response during a multiple-injection drug self administration session. PMID- 16220331 TI - The effects of a subanesthetic dose of ketamine on verbal memory in normal volunteers. AB - RATIONALE: N-methyl-D: -aspartate (NMDA) glutamate receptor antagonists have been reported to induce schizophrenia-like symptoms in humans, including memory impairments. Although the NMDA receptor has been shown to impair memory acquisition by disrupting long-term potentiation (LTP), limited research has been done on studying the effects of NMDA antagonists on the post-LTP cascade of events implicated in consolidation as measured by administering the drug after the initial learning experience. OBJECTIVE: The purpose of this experiment was to examine the effect of ketamine on mental status and to identify NMDA antagonist induced memory deficits by comparing the recall performance of items presented both immediately before and during ketamine infusion. METHODS: Thirteen normal controls received a 60-min infusion of ketamine in a randomized double-blind, cross-over design. Mental status was evaluated with the Brief Psychiatric Rating Scale and the Clinician-Administered Dissociative States Scale. The first 12-item word list was presented immediately before infusion, and two lists were subsequently presented during the infusion. Verbal memory performance was assessed by measuring the delayed cued recall of each list 30 min after its presentation. RESULTS: At the beginning, subjects experienced perceptual and reality distortion symptoms, followed later by mild subjective effects. Ketamine significantly reduced the delayed recall of words presented immediately before, but not during, drug infusion. Ketamine-induced decrements in verbal recall correlated significantly with plasma ketamine levels. CONCLUSION: This study characterizes the behavioral effects associated with ketamine and suggests that ketamine decreases verbal memory performance by interfering with early consolidation processes. PMID- 16220332 TI - MDMA (Ecstasy) and human dopamine, norepinephrine, and serotonin transporters: implications for MDMA-induced neurotoxicity and treatment. AB - RATIONALE: 3,4-Methylenedioxymethamphetamine (MDMA, designated as "Ecstasy" if illicitly marketed in tablet form) induces significant decrements in neuronal serotonin (5-HT) markers in humans, nonhuman primates, and rats as a function of dosing and dosing regimen. In rats, MDMA-mediated effects are attributed, in part, to selective high-affinity transport of MDMA into 5-HT neurons by the 5-HT transporter (SERT), followed by extensive 5-HT release. OBJECTIVES: To clarify whether SERT-selective effects of MDMA at human monoamine transporters can account for the reported MDMA-induced selective toxicity of serotonin neurons in primate brain. METHODS: We investigated the interaction of [(3)H](+/-, RS)- (+, S)- and (-, R)-MDMA with the human SERT, dopamine (DA) transporter (DAT), and norepinephrine (NE) transporter (NET) in stably transfected human embryo kidney (HEK)-293 cells. RESULTS: The human DAT, NET, and SERT actively transported [(3)H]RS(+/-)-MDMA saturably, stereoselectively, and in a temperature-, concentration-, and transporter-dependent manner. MDMA exhibited the highest affinity for the NET>>SERT>or=DAT, the same rank order for MDMA inhibition of [(3)H]DA, [(3)H]NE, and [(3)H]5-HT transport and stimulated release of the [(3)H]monoamines, which differed from reports derived from rodent monoamine transporters. The extent of MDMA-induced release of 5-HT was higher compared with release of DA or NE. CONCLUSIONS: The affinity of MDMA for the human SERT in transfected cells does not clarify the apparent selective toxicity of MDMA for serotonin neurons, although conceivably, its higher efficacy for stimulating 5-HT release may be a distinguishing factor. The findings highlight the need to investigate MDMA effects in DAT-, SERT-, and NET-expressing neurons in the primate brain and the therapeutic potential of NET or DAT inhibitors, in addition to SERT-selective inhibitors, for alleviating the pharmacological effects of MDMA. PMID- 16220333 TI - ACP-103, a 5-HT2A/2C inverse agonist, potentiates haloperidol-induced dopamine release in rat medial prefrontal cortex and nucleus accumbens. AB - RATIONALE: Atypical antipsychotic drugs (APDs) such as clozapine, olanzapine, quetiapine, risperidone, and ziprasidone are serotonin (5-HT)(2A) antagonists and relatively weaker dopamine (DA) D(2) antagonists, with variable 5-HT(2C) antagonist properties. The ability of atypical APDs to preferentially increase DA release in the cortex compared to the limbic system is believed to be due in part to their antagonism of 5-HT(2A) and D(2) receptors and believed to contribute to their beneficial effects on cognition, negative, and psychotic symptoms. Previous studies from this laboratory using microdialysis have shown that pretreatment of the 5-HT(2A) antagonist M100907 with the typical APD and D(2) antagonist haloperidol produced an increase in the medial prefrontal cortex (mPFC), but not in the nucleus accumbens (NAC), DA release. However, pretreatment with the 5 HT(2A/2C) receptor antagonist SR46349-B with haloperidol increased both mPFC and NAC DA release, suggesting that both 5-HT(2A) and 5-HT(2C) properties may be important for atypical APD effects. OBJECTIVE: The purpose of this study was to examine the effects of a novel putative atypical APD ACP-103 on mPFC and NAC DA release using in vivo microdialysis in freely moving rats that are awake. ACP-103 is an inverse agonist at both 5-HT(2A) and 5-HT(2C) receptors and has intermediate affinities for 5-HT(2C) receptors relative to their affinities for 5 HT(2A) receptors compared to M100907 and SR46349-B. In addition, the effects of ACP-103 were compared to M100907 and SR46349-B, and ACP-103 was also coadministered with haloperidol. RESULTS: ACP-103 10.0 mg/kg, but not 3.0 mg/kg, increased DA release in the mPFC, while neither dose increased DA release in the NAC. Like M100907, ACP-103 (3.0 mg/kg) potentiated 0.1 mg/kg haloperidol-induced DA release in the mPFC while inhibiting that in the NAC. However, ACP-103 (3.0 mg/kg), similar to SR46349-B, potentiated a high dose of haloperidol (1.0 mg/kg) induced DA release in both regions. The potent 5-HT(2C) antagonist SB242084 1.0 mg/kg significantly potentiated 0.1 mg/kg haloperidol-induced DA release in both the mPFC and NAC. However, SB242084, at 0.2 mg/kg, significantly potentiated DA release only in the NAC. Moreover, SB242084 0.2 mg/kg potentiated DA release in the NAC produced by the combination treatment of 3 mg/kg ACP-103 and 0.1 mg/kg haloperidol. CONCLUSION: These data suggest that the relative extent of 5-HT(2A) and 5-HT(2C) antagonism, as well as the extent of D(2) receptor blockade, has a critical influence on DA release in the mPFC and NAC and may be a determining factor in the action of this class of atypical APDs on these two potentially clinically relevant parameters. PMID- 16220334 TI - Strain-dependent antidepressant-like effects of citalopram in the mouse tail suspension test. AB - RATIONALE: Variations in the effects of antidepressant drugs between different mouse strains are important for drug discovery and could lead to the identification of genes that predict differences in drug efficacy. OBJECTIVES: This study compared behavioral baselines and dose-dependent responses to the selective serotonin reuptake inhibitor (SSRI) citalopram in eight inbred mouse strains (C57BL/6J, DBA/2J, C3H/HeJ, BALB/cJ, A/J, 129/SvEmsJ, 129/SvImJ, and BTBR) using the tail suspension test (TST). RESULTS: The DBA/2J, BALB/cJ, and BTBR strains were the most responsive to the effects of citalopram. Citalopram was least effective in the C57BL/6J and A/J strains. The antidepressant-like effects of citalopram in the TST were not correlated with changes in locomotor activity or deprivation-induced feeding behavior across the individual mouse strains, suggesting that patterns of sensitivity to citalopram are behaviorally specific and unlikely to result from pharmacokinetic variables. As an initial search for genetic polymorphisms causing differences in citalopram sensitivity, polymorphic forms of the tryptophan hydroxylase 2 (tph2) gene were genotyped and found to be not correlated with citalopram responsive (DBA/2J and BALB/cJ) and nonresponsive (A/J and C57BL/6J) strains. CONCLUSIONS: The TST strain survey described here: (1) suggested the most appropriate strains for screening potential antidepressants, (2) identified parental strains appropriate for quantitative trait loci mapping of genomic loci regulating SSRI sensitivity, and (3) indicated appropriate background strains for measuring an antidepressant-like response to the SSRI citalopram. The pattern of response agrees with a previous mouse strain survey that examined sensitivity to fluoxetine in the forced swim test (Lucki I, Dalvi A, Mayorga AJ (2001) Sensitivity to the effects of pharmacologically selective antidepressants in different strains of mice. Psychopharmacology 155:315-322). PMID- 16220336 TI - Effect of a novel nicotinic receptor antagonist, N,N'-dodecane-1,12-diyl-bis-3 picolinium dibromide, on nicotine self-administration and hyperactivity in rats. AB - RATIONALE AND OBJECTIVE: Recent work has shown that the novel compound N,N' dodecane-1,12-diyl-bis-3-picolinium dibromide (bPiDDB) may selectively block nicotinic acetylcholine receptors involved in regulating dopamine release. The current experiments examined the acute effect of bPiDDB on nicotine self administration, sucrose-maintained responding, and nicotine-induced changes in acute and sensitized locomotor activity. METHODS: Rats were first trained to respond for either nicotine (i.v.) or sucrose pellets using a standard two-lever operant conditioning procedure using a fixed ratio 5 schedule of reinforcement and were then pretreated with bPiDDB (0, 0.3, 1, or 3 mg kg(-1)) 15 min prior to the session. In separate experiments, rats were assessed for nicotine-induced changes in locomotor activity following pretreatment with bPiDDB (1 or 3 mg kg( 1)) or mecamylamine (1 mg kg(-1)); pretreatments were assessed with both acute and repeated nicotine (0.4 mg kg(-1)) treatment. RESULTS: Results showed that bPiDDB dose-dependently decreased nicotine self-administration, but not sucrose maintained responding. In the locomotor experiments, bPiDDB attenuated the hyperactivity produced by acute and repeated nicotine; however, this effect was not robust compared to mecamylamine. In contrast to mecamylamine, bPiDDB did not block the initial hypoactivity produced by acute nicotine. CONCLUSION: Since bPiDDB decreased nicotine self-administration specifically, this novel nicotinic receptor antagonist may constitute a lead for the development of a clinically useful treatment for tobacco dependence. PMID- 16220335 TI - Nicotinic effects on cognitive function: behavioral characterization, pharmacological specification, and anatomic localization. AB - RATIONALE: Nicotine has been shown in a variety of studies in humans and experimental animals to improve cognitive function. Nicotinic treatments are being developed as therapeutic treatments for cognitive dysfunction. OBJECTIVES: Critical for the development of nicotinic therapeutics is an understanding of the neurobehavioral bases for nicotinic involvement in cognitive function. METHODS: Specific and diverse cognitive functions affected by nicotinic treatments are reviewed, including attention, learning, and memory. The neural substrates for these behavioral actions involve the identification of the critical pharmacologic receptor targets, in particular brain locations, and how those incipient targets integrate with broader neural systems involved with cognitive function. RESULTS: Nicotine and nicotinic agonists can improve working memory function, learning, and attention. Both alpha4beta2 and alpha7 nicotinic receptors appear to be critical for memory function. The hippocampus and the amygdala in particular have been found to be important for memory, with decreased nicotinic activity in these areas impairing memory. Nicotine and nicotinic analogs have shown promise for inducing cognitive improvement. Positive therapeutic effects have been seen in initial studies with a variety of cognitive dysfunctions, including Alzheimer's disease, age-associated memory impairment, schizophrenia, and attention deficit hyperactivity disorder. CONCLUSIONS: Discovery of the behavioral, pharmacological, and anatomic specificity of nicotinic effects on learning, memory, and attention not only aids the understanding of nicotinic involvement in the basis of cognitive function, but also helps in the development of novel nicotinic treatments for cognitive dysfunction. Nicotinic treatments directed at specific receptor subtypes and nicotinic cotreatments with drugs affecting interacting transmitter systems may provide cognitive benefits most relevant to different syndromes of cognitive impairment such as Alzheimer's disease, schizophrenia, and attention deficit hyperactivity disorder. Further research is necessary in order to determine the efficacy and safety of nicotinic treatments of these cognitive disorders. PMID- 16220337 TI - Regulation of native GABAA receptors by PKC and protein phosphatase activity. AB - RATIONALE AND OBJECTIVE: Protein kinase C (PKC) modulation of ionotropic receptors is a common mechanism for regulation of channel function. The effects of PKC and phosphatase activation on native gamma-aminobutyric acid (GABA(A)) receptors in adult brain are unknown. Previous studies of recombinant GABA(A) receptors have provided evidence that PKC activation inhibits receptor function, whereas other studies suggest that PKC either increases or does not alter GABA(A) receptor function. The present study explored (a) the effects of PKC and phosphatase activity on GABA-mediated (36)Cl(-) uptake in cerebral cortical synaptoneurosomes and (b) the effect of PKC activity on muscimol-induced loss of righting reflex (LORR) in adult rats. METHODS: GABA(A) receptor function in vitro was measured by muscimol-induced (36)Cl(-) uptake into cerebral cortical synaptoneurosomes. The in vivo effect of PKC on GABA(A)-mediated function was measured by intracerebroventricular (i.c.v.) injection of 4-beta-phorbol-12,13 dibutyrate (PDBu) or calphostin C followed by determination of muscimol-induced LORR. RESULTS: Adenosine triphosphate (ATP) and PDBu produced a concentration dependent and specific reduction in muscimol-stimulated (36)Cl(-) uptake that was blocked by the PKC inhibitor calphostin C. Both adenosine diphosphate and 4alphaPDBu were ineffective. Phosphatase inhibition produced similar inhibition of muscimol responses. Furthermore, i.c.v. administration of PDBu and calphostin C produced opposing effects on both the onset and the duration of muscimol induced LORR in rats. CONCLUSIONS: The present study provides evidence that PKC activation reduces GABA(A) receptor function in native receptors both in vitro and in vivo. Phosphatase inhibitors decrease muscimol-mediated Cl(-) uptake in GABA(A) receptors demonstrating coordinated regulation of native receptors by PKC and phosphatases. PMID- 16220338 TI - Sensitization to amphetamine, but not PCP, impairs attentional set shifting: reversal by a D1 receptor agonist injected into the medial prefrontal cortex. AB - RATIONALE: Repeated exposure to psychomotor stimulants can lead to sensitization to their effects, and sensitization has been implicated in the pathophysiology of schizophrenia and drug abuse. These disorders are characterized by cognitive deficits, particularly in prefrontally mediated executive function. OBJECTIVE: The present experiments were conducted to investigate the effects of sensitizing regimens of amphetamine and phencyclidine (PCP) on attentional set shifting. METHODS: Rats received injections of amphetamine, PCP or saline three times per week for 5 weeks. Four weeks later, rats were trained to dig for food in one of two bowls, each bowl having an odour and a texture. Only one dimension (odour or texture) correctly predicted which bowl was baited. Rats were then tested on a series of discriminations including those requiring an intra-dimensional shift (IDS), an extra-dimensional shift (EDS) or a reversal of previously relevant and irrelevant stimuli. RESULTS: Rats sensitized to amphetamine performed normally on the IDS, but were impaired on the EDS, as well as on reversal discriminations. PCP-sensitized rats were unaffected on any of the discriminations. In amphetamine sensitized rats the deficit at the EDS stage was reversed by infusion of the D(1) receptor agonist SKF38393 into the medial prefrontal cortex (mPFC). CONCLUSIONS: Results show that the amphetamine-sensitized state impairs prefrontally mediated attentional set shifting. This is consistent with cognitive deficits in schizophrenia and addiction, and with the evidence that amphetamine sensitization is accompanied by functional changes in the mPFC. These results further add to a growing literature showing that activating D(1) receptors in the mPFC improves aspects of cognition. PMID- 16220339 TI - Chronic administration of the delta opioid receptor agonist (+)BW373U86 and antidepressants on behavior in the forced swim test and BDNF mRNA expression in rats. AB - RATIONALE: Selective delta opioid receptor agonists have been shown to produce antidepressant-like behavioral effects and increase brain-derived neurotrophic factor (BDNF) mRNA expression when given acutely, but the chronic effects of delta agonists have been less well characterized. OBJECTIVE: The present study examined the effects of chronic exposure to the delta agonist (+)BW373U86 (BW) on antidepressant-like behavior in the forced swim test and on BDNF mRNA expression in comparison to chronic treatment with the antidepressants fluoxetine, desipramine, bupropion, and tranylcypromine. METHODS: Sprague-Dawley rats were treated chronic ally with one of the above treatments and were tested for antidepressant effects in the forced swim test, and assayed for BDNF mRNA expression by in situ hybridization. RESULTS: Acute administration of 10 mg/kg BW produced a significant antidepressant-like effect in the forced swim test, while chronic (8- or 21-day) BW administration did not produce a significant antidepressant-like effect. When 10 mg/kg BW was administered for 8 days, it produced a significant increase in BDNF mRNA expression in the frontal cortex, while having no effect on BDNF expression when given for 21 days. Chronic bupropion and desipramine significantly decreased BDNF expression in the dentate gyrus of the hippocampus, while fluoxetine had no effect in any brain region. Chronic tranylcypromine produced a significant increase in BDNF expression in the CA1 region of the hippocampus. CONCLUSIONS: Chronic exposure to BW produces tolerance to most effects, although at differential rates. In addition, increased BDNF mRNA expression does not appear to be a common effect of chronic administration of various antidepressants. PMID- 16220340 TI - Inhibiting 5alpha-reductase in the amygdala attenuates antianxiety and antidepressive behavior of naturally receptive and hormone-primed ovariectomized rats. AB - RATIONALE: Greater incidence of anxiety and depressive disorders of women compared to men may be due in part to progesterone (P) and its neuroactive metabolite, 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP), acting in limbic regions, such as the amygdala. OBJECTIVE: If P's metabolism via 5alpha-reduction to 3alpha,5alpha-THP in the amygdala is critical for antianxiety and antidepressive behavior, then blocking 5alpha-reductase in the amygdala of female rats is likely to attenuate the antianxiety and antidepressive effects of high progestin levels from both endogenous and exogenous sources. METHODS: Naturally receptive female rats with high endogenous estrogen (E2) and P and ovariectomized (ovx) rats administered E2 (10 microg) and P (500 microg) subcutaneously were administered finasteride (10 microg/microl), a Type II 5alpha-reductase inhibitor, or vehicle to the amygdala. Anxiety behavior (open field, elevated plus maze, defensive freezing) and depressive behavior (Porsolt forced swim test) were assessed. RESULTS: There were similar effects of finasteride administration to the amygdala to attenuate antianxiety behavior in naturally receptive and ovx, hormone-primed rats. Finasteride administration significantly decreased central entries in the open field, decreased open arm time in the elevated plus maze, increased defensive freezing in response to footshock, and increased time spent immobile compared to vehicle. CONCLUSIONS: Thus, formation and subsequent actions of 3alpha,5alpha-THP in the amygdala may be important for antianxiety and antidepressive effects. PMID- 16220341 TI - [Neoadjuvant therapy for squamous cell carcinoma of the esophagus]. AB - Neoadjuvant treatment for locally advanced squamous cell carcinoma of the esophagus is widely used, despite minimal evidence of its efficacy. With only a minor effect on survival, the benefits of neoadjuvant radiotherapy alone appear to be limited. The same is true for neoadjuvant chemotherapy alone. While no single randomized study has shown a statistically significant result favoring neoadjuvant chemoradiation vs no treatment, the results of three meta-analyses indicate a prognostic benefit. Unfortunately, earlier chemoradiation protocols were associated with considerable side effects and contributed to perioperative morbidity and mortality. In contrast, modern chemoradiation protocols were revealed to be feasible when carried out in experienced institutions. Therefore, neoadjuvant chemoradiation is indicated in locally advanced stages. Patients should be referred to specialized centers for initial treatment planning and resection, and they should be enrolled in clinical studies whenever possible. PMID- 16220342 TI - Metabolomics and its role in understanding cellular responses in plants. AB - A natural shift is taking place in the approaches being adopted by plant scientists in response to the accessibility of systems-based technology platforms. Metabolomics is one such field, which involves a comprehensive non biased analysis of metabolites in a given cell at a specific time. This review briefly introduces the emerging field and a range of analytical techniques that are most useful in metabolomics when combined with computational approaches in data analyses. Using cases from Arabidopsis and other selected plant systems, this review highlights how information can be integrated from metabolomics and other functional genomics platforms to obtain a global picture of plant cellular responses. We discuss how metabolomics is enabling large-scale and parallel interrogation of cell states under different stages of development and defined environmental conditions to uncover novel interactions among various pathways. Finally, we discuss selected applications of metabolomics. PMID- 16220343 TI - Production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture. AB - The objective of this work was to produce doubled haploid plants from durum wheat through the induction of androgenesis. A microspore culture technique was developed and used to produce fertile doubled haploid plants of agronomic interest. Five cultivars, one selected line, plus a collection of 20 F(1) crosses between different genotypes of high breeding value were used. Studies on several factors such as pre-treatments and media components were carried out in order to develop a protocol to regenerate green haploid plantlets. Anthers were pre treated in 0.7 M mannitol. Microspores, from anther maceration, were plated on a C(17) induction culture medium with ovary co-culture. The optimum regeneration medium J25-8 was used. From 35 microspore isolations, 407 green plantlets were obtained. With this technique mature embryos were obtained. Green plants were regenerated from all genotypes used and approximately 67% of them were spontaneously doubled haploids. Some haploids and a very few polyploids plants were obtained. From the 407 plants, 275 were completely fertile and gave enough seeds to be assayed in the field. This protocol could be used complementary to or instead of the intergeneric crossing with maize as an economically feasible method to obtain doubled haploids from most durum wheat genotypes. PMID- 16220344 TI - Cloning and characterization of a novel splicing isoform of the iron-superoxide dismutase gene in rice (Oryza sativa L.). AB - Superoxide dismutases (SODs) are ubiquitous metalloenzymes in aerobic organisms that play a crucial role in protecting organisms against ROS. Here, we report the molecular cloning and functional characterization of a novel alternatively spliced variant of the iron-superoxide dismutase gene, OsFe-SODb, from a rice panicle cDNA library. The alternative splicing event occurred in the fourth exon of the OsFe-SOD gene, and led to the translation of two isoforms of different sizes. The 5' flanking region of the OsFe-SOD was cloned and many cis-acting regulatory elements were found that are involved in light responsiveness, including a G-box and an I-box. RT-PCR analysis showed that the two alternative forms of OsFe-SOD were expressed in both the vegetative and reproductive tissues of Cpslo17. Moreover, accumulation of both isoforms was upregulated by light induction. In addition, the alternative splicing of OsFe-SOD mRNA was sensitive to low temperature. High yield production of the two recombinant OsFe-SOD isoforms was achieved in Escherichia coli. SOD assays showed that C-terminal truncation in OsFe-SODb did not result in a loss of SOD enzyme activity. PMID- 16220345 TI - Nonsense mutation in TITF1 in a Portuguese family with benign hereditary chorea. AB - Benign hereditary chorea (BHC) is an autosomaldominant disorder of early onset characterized by a slowly progressing or nonprogressing chorea, without cognitive decline or other progressive neurologic dysfunction, but also by the existence of heterogeneity of the clinical presentation within and among families. The genetic cause of BHC is the presence of either point mutations or deletions in the thyroid transcription factor 1 gene (TITF1). We studied a Portuguese BHC family composed of two probands: a mother and her only son. The patients were identified in a neurology out-patient clinic showing mainly involuntary choreiform movements since childhood, myoclonic jerks, falls, and dysarthria. We performed magnetic resonance imaging (MRI), electroencephalogram (EEG), nerve conduction studies, thyroid ultrasound scan, biochemical thyroid tests, and electrocardiogram (ECG). We excluded Huntington disease by appropriate genetic testing and sequenced the entire TITF1 gene for both patients. The patients showed MRI alterations: (1) in the mother, abnormal hyperintense pallida and cortical cerebral/cerebellar atrophy; and (2) in the son, small hyperintense foci in the cerebellum and subtle enlargement of the fourth ventricle. Sequence analysis of the TITF1 gene in these patients revealed the presence of a heterozygous C > T substitution at nucleotide 745, leading to the replacement of a glutamine at position 249 for a premature stop codon. A previously undescribed nonsense mutation in the TITF1 gene was identified as being the genetic cause of BHC in this family. PMID- 16220347 TI - Intrahaplotype and interhaplotype pairing of bovine leukocyte antigen DQA and DQB molecules generate functional DQ molecules important for priming CD4(+) T lymphocyte responses. AB - Antigen-specific CD4(+) T-lymphocyte responses are restricted by major histocompatibility complex class II molecules, which influence T-cell priming during infection. Human leukocyte antigen (HLA) and bovine leukocyte antigen (BoLA) DRB3 and DQ genes are polymorphic, but unlike HLA, many BoLA haplotypes have duplicated DQ genes, and antibody-blocking studies indicated that BoLA-DQ molecules present various pathogen epitopes. Limited experimentation also suggested that BoLA-DQ molecules formed by interhaplotype pairing of A and B chains are functional. To compare antigen presentation by DR and DQ molecules and to definitively demonstrate functional BoLA-DQ molecules derived from interhaplotype pairing, different combinations of DR or DQ A and B proteins were expressed with CD80 in 293-F cells for use as antigen-presenting cells (APCs). This approach identified 11 unique restriction elements including five DR and six DQ pairs for antigen-specific CD4(+) T-cell responses against tick-transmitted bovine hemoparasites Anaplasma marginale or Babesia bovis. Interhaplotype pairing of DQ A and B molecules was demonstrated. Testing of six expressed DQA/B pairs from an animal with duplicated DQ haplotypes (DH16A/DH22H) demonstrated that an interhaplotype pair, DQA*2206/DQB*1301, presented A. marginale peptide B. In DH22H and DH16A homozygous animals, DQA*2206 was tightly linked with DQB*1402, and DQA*22021 was linked with DQB*1301. APCs from these donors could not present peptide B, confirming that DQA*2206/DQB*1301 encoded a functional interhaplotype pair. Functional BoLA-DQ molecules are generated by both intrahaplotype and interhaplotype pairing of A and B chains and play a similar role to BoLA-DR in priming helper T-cell responses to important pathogens. PMID- 16220346 TI - Replication of heterochromatin: insights into mechanisms of epigenetic inheritance. AB - Heterochromatin is composed of tightly condensed chromatin in which the histones are deacetylated and methylated, and specific nonhistone proteins are bound. Additionally, in vertebrates and plants, the DNA within heterochromatin is methylated. As the heterochromatic state is stably inherited, replication of heterochromatin requires not only duplication of the DNA but also a reinstallment of the appropriate protein and DNA modifications. Thus replication of heterochromatin provides a framework for understanding mechanisms of epigenetic inheritance. In recent studies, roles have been identified for replication factors in reinstating heterochromatin, particularly functions for origin recognition complex, proliferating cell nuclear antigen, and chromatin-assembly factor 1 in recruiting the heterochromatin binding protein HP1, a histone methyltransferase, a DNA methyltransferase, and a chromatin remodeling complex. Potential mechanistic links between these factors are discussed. In some cells, replication of the heterochromatin is blocked, and in Drosophila this inhibition is mediated by a chromatin binding protein SuUR. PMID- 16220349 TI - High re-bleeding rate in young adults after subarachnoid haemorrhage from giant aneurysms. AB - Low re-bleeding rates within the first 14 days of aneurysmal subarachnoid haemorrhage are reported in young patients. Furthermore, re-bleeding rate for giant aneurysms does not exceed 20% according to the literature. Our own clinical impression is that the re-bleeding rate seems to be much higher in giant aneurysms than reported, particularly in young patients. The aim of this study was to evaluate re-bleeding rate after subarachnoid haemorrhage following rupture of giant aneurysms in a younger population. We reviewed records of 23 patients who were treated in our institution for subarachnoid haemorrhage from giant aneurysms between 1994 and 2003. By definition, the aneurysms were larger than 25 mm in diameter. Five patients were younger than 40 years of age at the time of the aneurysmal subarachnoid haemorrhage. All younger patients (<40 years of age) showed re-bleeding after the first subarachnoid haemorrhage within the first 14 days of the initial event. In four patients (20%) older than 40 years at the time of the haemorrhage re-bleeding could be observed within the first 14 days of subarachnoid haemorrhage. We can confirm the re-bleeding rate of approximately 20% in patients suffering from subarachnoid haemorrhage (SAH) in the group of patients older than 40 years of age. However, younger patients seem to be at much higher risk of re-bleeding from giant aneurysms. PMID- 16220348 TI - Genomic characterization of MHC class I genes of the horse. AB - The availability of a contig of bacterial artificial chromosome (BAC) clones spanning the equine major histocompatibility complex (MHC) made possible a detailed analysis of horse MHC class I genes. Prior to this study, only a single horse MHC class I gene had been sequenced at the genomic level. Although many ( approximately 60) MHC class I cDNA sequences had been determined and published, from this information, it was not possible to determine how many class I loci are expressed in horses or to assign individual sequences to allelic series. In this study, 15 MHC class I genes were identified in BAC subclones and fully sequenced. Because the BAC library donor horse had been bred for homozygosity at the MHC, these 15 genomic clones represent distinct MHC class I genes and pseudogenes and not alleles at a smaller number of loci. For five of the genes, cDNA sequences from these loci had previously been identified. Two additional expressed class I genes were discovered, bringing the known total of different equine MHC class I genes (loci) expressed as mRNA to seven. Expression of all seven loci was detected by reverse transcriptase-polymerase chain reaction in adult, fetal, and placental tissues. The remaining eight genes were designated as pseudogenes. This work resulted in moderate expansion of the horse MHC BAC contig length, and the remaining gap was shortened. The information contained in these equine MHC class I sequences will permit comparison of MHC class I genes expressed across different horse MHC haplotypes and between horses and other mammalian species. PMID- 16220350 TI - Intraventricular meninigiomas: a report of 25 cases. AB - In order to study the clinical characteristics of intraventricular meningiomas, we analyzed retrospectively 25 patients and reviewed the literature with regard to incidence, clinical manifestation, imaging features, preoperative diagnosis, surgical findings, and histopathological results. Intraventricular meningiomas are quite rare, but they represent an important differential diagnosis of intraventricular neoplasms. Computed tomography or magnetic resonance imaging enable a correct diagnosis of intraventricular meningiomas in most of the cases. The tumors often grow slowly to a substantial size before they become symptomatic. The operative route should be selected according to the tumor's location. Out of the 24 lateral ventricular meningiomas in our series, 20 were resected via a posterior parieto-occipital transcortical approach, two were resected via a transcallosal approach, and another two tumors, located in the frontal horn and body of the lateral ventricle, were resected via a frontal approach. A median suboccipital craniotomy was performed for the fourth ventricular meningioma. The parieto-occipital route for lateral ventricular meningiomas is a safe surgical approach, which is not necessarily associated with postoperative visual deficits. Piecemeal removal of the tumor can be safely and easily performed and special attention should be paid to the choroidal vessels intraoperatively. PMID- 16220351 TI - Spinal canal narrowing during simulated frontal impact. AB - Between 23 and 70% of occupants involved in frontal impacts sustain cervical spine injuries, many with neurological involvement. It has been hypothesized that cervical spinal cord compression and injury may explain the variable neurological profile described by frontal impact victims. The goals of the present study, using a biofidelic whole cervical spine model with muscle force replication, were to quantify canal pinch diameter (CPD) narrowing during frontal impact and to evaluate the potential for cord compression. The biofidelic model and a sled apparatus were used to simulate frontal impacts at 4, 6, 8, and 10 g horizontal accelerations of the T1 vertebra. The CPD was measured in the intact specimen in the neutral posture (neutral posture CPD), under static sagittal pure moments of 1.5 Nm (pre-impact CPD), during dynamic frontal impact (dynamic impact CPD), and again under static pure moments following each impact (post-impact CPD). Frontal impact caused significant (P<0.05) dynamic CPD narrowing at C0-dens, C2-C3, and C6-C7. The narrowest dynamic CPD was observed at C0-dens during the 10 g impact and was 25.9% narrower than the corresponding neutral posture CPD. Interpretation of the present results indicate that the neurological symptomatology reported by frontal impact victims is most likely not due to cervical spinal cord compression. Cord compression due to residual spinal instability is also not likely. PMID- 16220355 TI - Performance of 2-deoxy-2-[F-18]fluoro-D-glucose positron emission tomography and integrated PET/CT in restaged breast cancer patients. AB - PURPOSE: This study was conducted to compare the clinical stage derived from 2 deoxy-2-[F-18]fluoro-D-glucose (FDG) positron emission tomography (PET) to that of integrated PET/computed tomography (CT) in restaged breast cancer patients. PROCEDURES: Fifty-eight female patients (age range 29-80 years, mean age +/-SD, 53.3 +/- 11.7 years) underwent PET/CT restaging for breast cancer. Two experienced nuclear medicine physicians interpreted PET images. A radiologist was added for reading PET/CT studies. A patient-based analysis was performed. Histopathological findings, correlative imaging studies, changes in number, size, and hypermetabolic activity of suspicious lesions and/or patient outcome served as standard of reference for determining the diagnostic accuracy of both modalities. RESULTS: PET staged 79.3% (46/58) of the patients correctly, overstaged seven (12.1%), and understaged five patients (8.6%). Integrated PET/CT staged 89.7% (52/58) of the patients correctly, overstaged four (6.9%), and understaged two patients (3.4%). The staging accuracy of PET/CT was not significantly better than that of PET alone (p = 0.059). Lesions exhibiting mild hypermetabolic activity, benign inflammatory lesions, and physiological variants largely explained incorrect PET findings. CONCLUSION: Integrated PET/CT only marginally improves the restaging accuracy over PET alone (p = 0.059) in breast cancer patients. PMID- 16220354 TI - Mouse extrahepatic hepatoma detected on MicroPET using copper (II)-64 chloride uptake mediated by endogenous mouse copper transporter 1. AB - PURPOSE: This study was conducted to assess the positron-emitting copper (II)-64 chloride ((64)CuCl(2)) as a probe for imaging mouse extrahepatic hepatoma expressing mouse copper transporter 1 (mCtr1) with positron emission tomography (PET). PROCEDURES: Following the intravenous administration of (64)CuCl(2), athymic mice bearing extrahepatic hepatoma grafts were subjected to whole-body static PET imaging with a Concorde microPET R4 tomograph. Upon completion of the imaging study, immunohistochemistry (IHC) study of mCtr1 was performed with postmortem tissues. RESULTS: The mouse extrahepatic hepatoma grafts were well visualized on static microPET images. Quantitative analysis demonstrated that the tracer concentration in the hepatoma was significantly higher than those in the soft tissue of the right shoulder opposite to the tumor site and the brain (p < 0.001). mCtr1 immunoreactivity in the hepatoma graft was approximately 70% of that in liver, whereas (64)CuCl(2) concentration in the graft was approximately 11% of the liver concentration. CONCLUSIONS: The extrahepatic mouse hepatoma grafts may be visualized by Cu-64 PET, taking advantage of the (64)CuCl(2) uptake mediated by the functional endogenous mCtr1. PMID- 16220357 TI - Behavioral response in the terrestrial isopod Porcellio scaber (Crustacea) offered a choice of uncontaminated and cadmium-contaminated food. AB - The objective of this study was to find out whether Porcellio scaber discriminates against Cd-contaminated food. The foraging behavior in animals offered uncontaminated and Cd-contaminated food simultaneously was quantified for 48-h employing computer-aided video tracking. To see whether the isopods' selection of less contaminated food could diminish the influence Cd on food consumption, growth, metal assimilation, moulting and mortality, Cd-dosed food (20, 45, 200 and 450 mg kg(-1) dry weight) was offered together with untreated food for 3 weeks. Data from the video tracking experiments revealed that animals visited Cd-dosed food as often as untreated food, but spent much less time near Cd-dosed food. Discrimination against Cd-contaminated food increased with previous experience with contaminated food and/or with increased Cd body burden. In 3 weeks exposure uncontaminated food preference rose with time of exposure and cadmium concentration in food and reached a maximal preference ratio of 65% (untreated food): 35% (Cd-dosed food). The decreased consumption of Cd-dosed food was compensated by the increased consumption of control food. Cadmium body burden increased with time of exposure and cadmium concentration in food consumed, while the influence of Cd on food consumption, growth and moulting was diminished. PMID- 16220356 TI - Predictors of smoking cessation in pregnancy and maintenance postpartum in low income women. AB - OBJECTIVE: To describe factors associated with smoking status of low-income women during pregnancy and postpartum. METHODS: Data from a randomized clinical trial were used to conduct separate analyses on 327 women who smoked at baseline (time at enrollment) and for whom smoking status was available at delivery, and on 109 women who reported not smoking at delivery (quit spontaneously or after study enrollment) and for whom smoking status was available at 6-months postpartum. Salivary cotinine was used to assess the accuracy of self-reported smoking status for the sample as a whole. Data were collected between May 1997 and November 2000. RESULTS: 18% of the 327 baseline smokers stopped smoking before delivery. Cessation was less likely in older women, those reporting Medicaid coverage (vs. commercial or no insurance), who were at a later week of pregnancy at baseline, were more addicted, had a husband/partner who smoked, and did not receive the study intervention. 37% of the 109 women who reported not smoking at delivery maintained abstinence at 6-months postpartum. Factors associated with abstinence were later week of pregnancy at baseline and quitting spontaneously with pregnancy, while women who lived with a smoker were less likely to report abstinence. Spontaneous quitters were less likely to relapse by 6 months postpartum than women who quit smoking later in pregnancy. CONCLUSIONS: Partner participation in smoking cessation programs for pregnant and postpartum women merits exploration. Lower relapse rates among spontaneous quitters indicate a need to foster an environment that encourages quitting at pregnancy. PMID- 16220358 TI - Short- and long-term response of sunflower to airborne bromoxynil-octanoate under controlled and field conditions. AB - Wind tunnel studies and a field study were conducted to investigate the impact of the airborne herbicide bromoxynil-octanoate on non-target plants. Sunflowers at the two-leaf stage were used as test plants and exposed for 24 h in a wind tunnel to a range of concentrations of airborne bromoxynil-octanoate. Quantum yield of electron transport at Photosystem II (phiPSII) of exposed leaves and leaves developed after exposure, plant height, days to flower and head diameter were used to determine the short- and long-term response of sunflowers to bromoxynil octanoate. Quantum yield of exposed leaves and of the second leaf pair, determined 1 h and 2 days after exposure, was reduced at concentrations >0.449 and >1.159 microg/m3, respectively, but no effects on further leaf pairs developed after exposure were observed. Growth parameters were partly affected when exposed to >1.448 microg/m3. In a field study a maize plot was treated with bromoxynil-octanoate and parathion-methyl, used as a tracer compound. Afterwards, potted sunflowers were placed inside the maize plot and at different distances (2, 4, 8 and 16 m) from the treated plot. The highest concentrations of bromoxynil-octanoate and parathion-methyl collected in air samples at a distance of 8 m outside of the treated plot were 0.100 and 0.055 microg/m3, respectively. Little of the breakdown product bromoxynil phenol was detected in the air. Detectable concentrations of bromoxynil-octanoate and bromoxynil phenol were found only in a few leaf samples, whereas parathion-methyl was detected in almost 50% of the leaf samples up to 16 m from the treated plot. Quantum yield and growth of sunflower were not affected due to low concentrations of bromoxynil octanoate in the air. PMID- 16220359 TI - Chromosomal damage in two species of aquatic turtles (Emys orbicularis and Mauremys caspica) inhabiting contaminated sites in Azerbaijan. AB - The Caspian region and specifically the Apsheron peninsula of Azerbaijan are known to be polluted with a variety of environmental contaminants. These complex mixtures of contaminants make risk assessment difficult. We used the flow cytometry method (FCM) and the micronucleus assay (MN) to assess chromosomal damage in aquatic turtles (Emys orbicularis, the European pond turtle; and Mauremys caspica, the Caspian turtle) inhabiting contaminated wetlands in Azerbaijan. Evidence of genetic damage was found for two sites, Neftchala and Sumgayit, relative to a reference site, Ali Bairamly. Sediment samples from each site were analyzed for PAHs and mercury to evaluate potential contaminant associations with genetic damage. A significant positive correlation was documented between three-ring PAH sediment concentrations and FCM estimates of chromosomal damage in E. orbicularis. These data combine to show that the contaminated wetlands in Sumgayit and Neftchala are genotoxic and that three-ring PAHs are likely a significant influence on observed genotoxicity. PMID- 16220361 TI - Ecotoxicological and fertilizing effects of dewatered, composted and dry sewage sludge on soil mesofauna: a TME experiment. AB - The effects of dewatered, composted and dry urban sewage sludge on the soil mesofauna were tested in mesocosms. PVC containers were filled with soil/sludge mixtures in a proportion to amount to 6% organic matter content and were colonized with soil fauna coming from undisturbed forest soils. Mesocosms were incubated under laboratory conditions for 7, 30, 60, 120 and 180 days, after which fauna was extracted in Berlese funnels. The animals were classified at different taxonomic levels. Acari were classified to the suborder level for Astigmata, to the family level for Mesostigmata and Prostigmata and to the species level for Cryptostigmata. Acute- and medium-term effects were determined on the faunal density, relative abundance of the main taxa and community structure. Prostigmata were sensitive to the acute effect of the sludge, whereas Mesostigmata and particularly Cryptostigmata were sensitive to its medium-term effect. The most negative effects were found for dry sludge, which caused acute and medium-term effects on the invertebrate communities and on the soil trophic structure. PMID- 16220360 TI - DNA strand length and EROD activity in relation to two screening measures of genotoxic exposure in Great Lakes herring gulls. AB - We collected tissues from herring gulls (Larus argentatus) nesting within and outside of the Great Lakes basin. Genotoxin exposure was assessed as fluorescent aromatic compounds (FACs) in bile and SOS Chromotest-inducing activity in muscle extracts. We determined whether these exposures were associated with decreased erythrocyte DNA strand length and/or induction of hepatic ethoxyresorufin-O deethylase (EROD) activity. FACs were detected in all bile samples. Most muscle extracts produced a positive or marginal SOS response in the presence of S9. SOS induction potentials were strongly associated with dietary trophic level. The median molecular length of DNA isolated from erythrocytes for 14 of 17 adult and 10 of 11 prefledgling collections was reduced compared to the modal class for their respective age group suggesting widespread DNA damage. DNA damage was greatest in gulls from Saginaw Bay, Lake Huron. Median EROD activity in both adults and prefledglings from remote locations was significantly lower than that of gulls from the lower Great Lakes and was not associated with concentrations of benzo[a]pyrene (B[a]P)-like FACs. Our results indicate Great Lakes herring gulls were exposed to genotoxins and Ah-receptor activating agents in biologically significant concentrations in the early 1990s. These agents appear to be persistent bioaccumulative compounds and/or their metabolites. PMID- 16220362 TI - Effects of the herbicide isoproturon on metallothioneins, growth, and antioxidative defenses in the aquatic worm Tubifex tubifex (Oligochaeta, Tubificidae). AB - Metallothioneins (MTs) are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MT contents is considered to be a specific biomarker of metal exposure. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth, and antioxidative defenses. Therefore, the induction of MTs as biomarkers of exposure to the pesticide isoproturon has been investigated in the aquatic worms Tubifex tubifex. MT levels in exposed worms increased significantly (p < 0.05) after 2, 4, and 7 days of exposure to different concentrations of isoproturon (maximum increase compared to unexposed controls: +148.56% for 10 mg l(-1) after 4 days of exposure). In response to isoproturon, the activity of glutathione-S-transferase (max. +52%), glutathione-reductase (max. +100%), and catalase (max. +117%) increased, demonstrating the occurrence of an oxidative stress response to the herbicide. Thus, the increase in MT contents caused by isoproturon was interpreted as a defense response towards increased oxidative stress generated by the herbicide. Residues of isoproturon and its metabolites, 1-(4-isopropylphenyl) 3-methylurea, 1-(4-isopropylphenyl) urea, and 4-isopropylanilin were detected in the worm growth medium. Half-life of the herbicide was shorter at a low (0.1 mg l(-1)) initial concentration. The herbicide accumulated in T. tubifex but no metabolite could be detected. PMID- 16220363 TI - Influence of non-point source pollution on riverine fish assemblages in South West France. AB - The relationship between non-point source pollution (NSP) and fish assemblages in the Garonne basin, SW France was studied. Two independent data sets were coupled, one containing 20 physico-chemical variables and another containing 40 fish species in 84 study sites. Species were classified in guilds according to their feeding habitat and their diet composition. The physico-chemical variables were log-transformed and standardized for a factor analysis in which they were grouped into four factors which accounted for 80% of the total variability. These were named according to factor loadings (i.e. a measure of the variance of a given variable) whose absolute values were larger than 0.5. Hence, the first factor (F1) was formed by variables linked to NSP, most notably by sodium, chloride, potassium, orthophosphates, nitrites and chemical oxygen demand. The second factor (F2) was related to alkalinity (i.e. bicarbonates, calcium, conductivity and pH). The third factor (F3) included oxygen saturation rate and dissolved oxygen, and F4 combined both temperature and flow. Factor scores (i.e. weighted sums of the original variables) were then introduced in stepwise multiple regression models as explanatory variables of log-transformed fish species richness of trophic guilds. The NSP factor was significant (p < 0.05) for the following models: benthic omnivores (r2 = 0.66), all species (r2 = 0.65), total benthic species (r2 = 0.63), total water-column species (r2 = 0.57), benthic invertivores (r2 = 0.32) and water-column invertivores (r2 = 0.16). The guilds for which NSP was not significant were water-column omnivores, water-column piscivores and benthic detritivores. Thus, there was evidence of an inversely proportional association, though not causation, of NSP with species richness of riverine fish trophic guilds on a large spatio-temporal scale. Fish assemblages may respond in different ways to NSP depending on their species composition, on the region and on the scale, and not only to physico-chemical properties of water. PMID- 16220364 TI - Identification of sex chromosome molecular markers using RAPDs and fluorescent in situ hybridization in rainbow trout. AB - The goal of this work is to identify molecular markers associated with the sex chromosomes in rainbow trout to study the mode of sex determination mechanisms in this species. Using the RAPD assay and bulked segregant analysis, two markers were identified that generated polymorphic bands amplifying preferentially in males of the Mount Lassen and Scottish strains of rainbow trout. Chromosomal localization using fluorescent in situ hybridization of a 900 bp probe developed from one of these markers revealed a brightly defined signal on a chromosome that could morphologically be classified as the Y chromosome. PMID- 16220366 TI - Isozymes in Larrea divaricata and Larrea tridentata (Zygophyllaceae): a study of two amphitropical vicariants and autopolyploidy. AB - Electrophoretic variants for seven isozyme systems - probably encoded by 18 structural gene loci - in diploid populations of Larrea divaricata and diploid and tetraploid populations of its North American vicariant derivative L. tridentata were assayed by polyacrilamide and starch gel electrophoresis. High molecular similarity of diploid and tetraploid cytotypes of L. tridentata supports the hypothesis of interracial autopolyploidy. The absence of fixed heterozygosity and additive profiles indicates a low level of divergence between the parental diploids and the tetraploids. The phenogram based on the I coefficient showed the similarities between the populations of diploid L. divaricata and also between the diploid populations of L. tridentata. Both groups of diploid populations were more distantly connected to tetraploid L. tridentata. PMID- 16220367 TI - Old age, multiple formations or genetic plasticity? Clonal diversity in the uniparental Caucasian rock lizard, Lacerta dahli. AB - Allozyme variation at 35 gene loci is investigated in 161 specimens of the uniparental Caucasian lizard Lacerta dahli from several locations in Armenia and Georgia. All individuals are heterozygotic at 12 loci, and homozygotic at 21 loci. Variation at two loci results in five uniparental clones. One clone is widespread whereas four are geographically restricted and are represented by only one or two individuals. Because successful formation of uniparental clones is rare, and because the biparental species forming them are now allopatric, the most probable explanation for the origin of the observed clonal diversity is either mutation or recombination within the common clone. The rare clones have lower levels of enzyme activity at four loci, suggesting that these organisms may be genetically deficient. Although the evidence points to change in a pre existing clone, the possibility of multiple origins cannot be ruled out. PMID- 16220369 TI - Patterns of DNase I sensitivity in the chromosomes of the ant Tapinoma nigerrimum (Hymenoptera, Formicidae). AB - We have analysed the patterns of DNase I/nick translation in the chromosomes of Tapinoma nigerrimum. The results show a non-uniform DNase I sensitivity in different chromosome domains. The hypersensitivity appears to be specially concentrated at both the NOR and the distal regions. The resemblance to and differences from the situation in other animal species, in which active genes are DNase I hypersensitive, are discussed. PMID- 16220368 TI - Isozyme variation in Larrea ameghinoi and Larrea nitida (Zygophyllaceae): genetic diversity and its bearing on their relationship. AB - Electrophoretic variants for 11 isozyme systems were examined by horizontal polyacrilamide gel electrophoresis in natural populations of Larrea ameghinoi and L. nitida (section Larrea). The two taxa presented fixed alternative allelic variants at loci Adh-1, Gdh-1 and Mdh-1. Genetic variability estimates showed no statistically significant differences among populations of section Larrea (He: 0.097-0.167). However, these values were significantly lower than those reported for species of section Bifolium (L. divaricata, diploid L. tridentata) in a previous contribution (He: 0.17-0.29). Positive and highly significant Wright's fixation indices are in agreement with higher amounts of inbreeding for L. nitida and L. ameghinoi, as suggested on the basis of floral morphology by previous authors. Moreover, the higher levels of genetic differentiation obtained for conspecific populations of section Larrea as compared to those of section Bifolium are also consistent with these observations. Nei's genetic identity values obtained for sympatric (I = 0.80) and allopatric (I = 0.63-0.73) L. ameghinoi - L. nitida population pairs are concordant with those expected for different, but closely related congeneric species. PMID- 16220371 TI - Unusual occurrence of a ZZ/ZW sex-chromosome system and supernumerary chromosomes in Characidium cf. fasciatum (Pisces, Characiformes, Characidiinae). AB - Individuals of two populations of the fish Characidium cf. fasciatum were cytogenetically studied and showed a basic diploid number of 50 chromosomes. Some fishes were found to have 51 to 54 chromosomes due to the presence of one to four small subtelocentric/acrocentric supernumerary chromosomes. When analyzed by conventional Giemsa staining, male and female specimens of C. cf. fasciatum from the Quinta stream and Pardo River presented the same basic karyotypic macro- and microstructure, consisting of 32 metacentric and 18 submetacentric chromosomes. Ag-NORs were terminally located on the long arms of two submetacentric chromosome pairs. Constitutive heterochromatin was identified by C-banding as small pericentromeric blocks in the majority of the chromosomes, and B-chromosomes were found to be heterochromatic. The occurrence of one totally heterochromatic submetacentric chromosome restricted to females and considered as an unusual feature in fish karyotypes led to the identification of a ZZ/ZW sex-chromosome system. The implications of chromosomic differentiation observed in the genus Characidium are discussed. PMID- 16220372 TI - Genetic differentiation in Spanish populations of Ceratitis capitata as revealed by abundant soluble protein analysis. AB - Ceratitis capitata is one of the most important pest species in the tropical and temperate regions of the world, however, genetic knowledge of this species is still very limited. In the present study, we have attempted for the first time an analysis of the genetic variability in seven natural populations of C. capitata by means of abundant soluble proteins, combining high resolution techniques such as polyacrylamide gel electrophoresis and silver staining methods. A total of 66 polypeptides have been analyzed. The results show the existence of a decreasing trend in the 'levels' of polymorphism from the southern areas to the northern that is correlated with the latitude of the sampling areas, most probably due to the colonization process of the Iberian Peninsula. On the other hand, and with relation to the 'pattern' of the variability, a geographic differentiation (not strictly latitudinal) is detected when an UPGMA clustering method was applied to the data. Moreover, principal component anaysis has revealed that a part of this differentiation could be explained in relation with environmental factors such as annual rainfall and minimum temperature in winter months. This is suggesting that selection (added to the historical process) could be playing an important role in the process of geographic differentiation, 'shaping' the pattern of that variability. No host-related differentiation has been observed in these samples. PMID- 16220373 TI - Assessing genetic diversity in a sugarcane germplasm collection using an automated AFLP analysis. AB - An assessment of genetic diversity within and between Saccharum, Old World Erianthus sect. Ripidium, and North American E.giganteus (S.giganteum) was conducted using Amplified Fragment Length Polymorphism (AFLP(TM)) markers. An automated gel scoring system (GelCompar(TM)) was successfully used to analyse the complex AFLP patterns obtained in sugarcane and its relatives. Similarity coefficient calculations and clustering revealed a genetic structure for Saccharum and Erianthus sect. Ripidium that was identical to the one previously obtained using other molecular marker types, showing the appropriateness of AFLP markers and the associated automated analysis in assessing genetic diversity in sugarcane. A genetic structure that correlated with cytotype (2n=30, 60, 90) was revealed within the North American species, E. giganteus (S.giganteum). Complex relationships among Saccharum, Erianthus sect. Ripidium, and North American E.giganteus were revealed and are discussed in the light of a similar study which involved RAPD markers. PMID- 16220374 TI - Maternal effect of a hybrid inviability gene in Tribolium castaneum. AB - The Tribolium castaneum hybrid inviability gene, H, was selectively introgressed into a genetic background lacking H through serial paternal backcrosses. This revealed a poor viability phenotype (partial paralysis and poor control of the limbs, referred to as tremor) not present in the parent strains. Tremor cosegregated with H, but was expressed only when transmitted paternally and only when H was not also present maternally. The inferred maternal, self-suppressive effect of H may explain nonreciprocal incompatibilty previously observed between H and H-incompatible strains. PMID- 16220375 TI - One step beyond lethal equivalents: characterization of deleterious loci in the rapid cycling Brassica rapa L. base population. AB - The total number of lethal equivalents as defined by Morton, Crow and Muller (1956) is a function of three parameters: M, the number of loci at which deleterious mutations can occur, q, the frequency of the deleterious alleles at each locus, and s, their selective value. A new approach based on multi generation inbreeding data is outlined and used to infer these three parameters as well as the dominance coefficient, h, in a self-incompatible species, Brassica rapa L. Germination and flowering data from thirty bud-selfed lines of fast cycling B.rapa were assessed over three generations. Germination and flowering were significantly postponed by inbreeding but germination and flowering success were not so strongly decreased. Estimates of the average s values were obtained but it was not possible to get separate estimates of M and q. For both characters, the average dominance coefficient was particularly low. The number of lethal equivalents at the zygotic level was around two for germination and three for flowering, which, owing to the self-incompatibility of B.rapa, is an unexpectedly low value. These results may be explained by past biparental inbreeding which in turn may have increased self-compatibility thus allowing the purging of more deleterious alleles than under strict self-incompatibility. PMID- 16220376 TI - RAPD technique for taxonomic studies of Pellia epiphylla-complex (Hepaticae, Metzgeriales). AB - The random amplified polymorphic DNA (RAPD) technique was used to study the genomic relationships of three liverworts from the genus Pellia: P.epiphylla, P. borealis and P. neesiana. Altogether 150 characters (150 DNA fragments obtained using PCR) were scored. These characters were used to create a matrix of pairwise distances between all the pairs of taxa. Both distance (UPGMA, Fitch-Margoliash and Neighbor-Joining) and binary character-state (Wagner and Camin-Sokal parsimony and compatibility) methods were applied for trees' construction. Our results strongly support distinction of the recently discovered sibling species of P.epiphylla - species N and P.epiphylla - species S, which have an allopatric distribution in Poland (N - North, S - South Poland). Moreover, our data also supports the hypothesis of a hybrid origin (alloploid) of the polyploid P. borealis from P. epiphylla-NxP.epiphylla-S. P.neesiana was excluded as a donor of either of the genomes of P. borealis. PMID- 16220378 TI - Present knowledge of kin discrimination in salmonids. PMID- 16220390 TI - Incompatibility and pollen competition in Alnus glutinosa: Evidence from pollination experiments. AB - Different types of incompatibility systems were found to operate simultaneously in Alnus glutinosa in the course of numerous pollination experiments, including self-pollination and pollination with controlled pollen mixtures. Isozyme genetic markers were used to identify the pollen parent of each offspring from the mixed pollination experiments, thus allowing specification of the fertilization success of each pollen parent. In a first step, these results were compared with observations on in vitro pollen germination experiments. This comparison allows for exploration of the explanatory value of different germination media as models of germination conditions on stigmas. In most cases, the data suggest that the in vitro germination conditions resemble the fertilization conditions in vivo, at least in the sense that they favor the same pollen parents. By providing a generic and operable definition of the two basic types of incompatibility, eliminating (inability to fertilize ovules) and cryptic (resulting in lowered fertilization success of a pollen parent under competition), evidence was detected for the existence of both types of incompatibility in Alnus glutinosa, where eliminating incompatibility occurred as self-incompatibility only. However, since this incompatibility seems to act primarily via pollen elimination, seed production is not likely to be negatively affected in natural populations, even for comparatively large amounts of self-pollination. PMID- 16220393 TI - RAPD divergence caused by microsite edaphic selection in wild barley. AB - Random amplified polymorphic DNA polymerase chain reaction (RAPDPCR) was used to assess genetic diversity in four subpopulations (86 individuals) of wild barley, Hordeum spontaneum, sampled from Tabigha microsite near the Sea of Galilee, Israel. The microsite consists of two 100 m transects that are topographically separated by 100 m, each equally subdivided into 50 M of basalt and terra rossa soil types. Despite the same macroclimate characterizing the area around the Sea of Galilee, the microsite offers two edaphically different microhabitats, with basalt being a more ecologically heterogeneous and broaderniche than the relatively drier but more homogeneous and narrowniche terra rossa. Analysis of 118 putative loci revealed significant (P<0.05) genetic differentiation in polymorphism (P0.05) between the two soils across the transects with P being higher in the more heterogeneous basalt (mean P0.05 = 0.902), than in terra rossa (mean P0.05 = 0.820). Gene diversity (He) was higher in basalt (mean He=0.371), than in terra rossa (mean He=0.259). Furthermore, unique alleles were confined to one soil type, either in one or both transects. Rare alleles were observed more frequently in terra rossa than basalt, and in transect II only. Gametic phase disequilibria showed a larger multilocus association of alleles in basalt than terra rossa, and in transect I than II. Spearman rank correlation (r(s)) revealed a strong association between specific loci and soil types, and transects. Also, analysis of multilocus organization revealed soilspecific multilocusgenotypes. Therefore, our results suggest an edaphically differentiated genetic structure, which corroborates the niche widthvariation hypothesis, and can be explained, in part, by natural selection. This pattern of RAPD diversity is in agreement with allozyme and hordein protein diversities in the same subpopulations studied previously. PMID- 16220394 TI - Genetic differentiation in Thai populations of the rare species Afgekia sericea Craib (Leguminosae) revealed by RAPDPCR assays. AB - Genetic diversity within and among populations was investigated using RAPD-PCR assays in a rare species, Afgekia sericea Craib. Two hundred and sixty-nine individuals were sampled from nine geographically isolated populations from northeastern Thailand. This study includes 73 RAPD markers. Within population polymorphism as measured by percentage of polymorphic RAPDs, varied between 38.4% and 60.3%. Genetic variability was measured using Shannon's information index and partitioned into between- and within-population components. Overall, genetic variation among A. sericea populations was high (H(sp)-H(pop))/H(sp)=48.2%. The genetic diversity for the species (H(sp)) was 43.2%. Mean of within-population values (H(pop)) for all populations was 0.224. The total genetic diversity was explained by high variation among populations (mean Gst=0.426), which is consistent with low gene flow among populations (Nm=0.35). High between population genetic variation observed in this study could be explained by limited migration through seed and/or pollen dispersal among populations. Conservation strategies of A. sericea are discussed in the context of these results. PMID- 16220396 TI - Retrolyc1-1, a member of the Tntl retrotransposon super-family in the Lycopersicon peruvianum genome. AB - Retrotransposons are ubiquitous mobile genetic elements that transpose through an RNA intermediate. One of the best known plant retrotransposon, Tnt1, was isolated from tobacco and showed an extensive distribution in the Nicotiana genus. We investigated the presence of related sequences in the Lycopersicon genus, another member of the Solanaceae family. Hybridization experiments performed using Tnt1 probes indicated that homologous sequences were present in all Lycopersicon species, indicating that these Tnt1-related sequences, that we named Retrolyc1, are distributed throughout the Lycopersicon genus. Different distribution patterns were detected between species, demonstrating a potential use of Retrolyc1 elements as molecular markers. An incomplete Retrolyc1 sequence, that we named Retrolyc1-1, was isolated from an L. peruvianum genomic library. Retrolyc1-1 shows extensive homology with Tnt1 sequences except in the LTR U3 region. Since this region is known to be involved in the control of transcription, this strongly suggests the existence of different patterns of regulation for Tnt1 and Retrolyc1 elements. The study of these two elements within the Solanaceae family may provide interesting models for retrotransposon evolution within this group and transmission in host genomes. PMID- 16220400 TI - Late effects and cosmetic results of conventional versus hypofractionated irradiation in breast-conserving therapy. AB - BACKGROUND AND PURPOSE: Breast irradiation after lumpectomy is an integral component of breast-conserving therapy (BCT). As the prognosis is general good following BCT, late morbidity and cosmesis are important. The present study compares two different radiation schedules with respect to these two endpoints. PATIENTS AND METHODS: 129 breast cancer patients (pT1-2 pN0-1 cM0) were irradiated between 09/1992 and 08/1994 with either a 22-day fractionation schedule (2.5 Gy to 55 Gy, 4x/week, n = 65) or with a conventional fractionation schedule (28 days, 2.0 Gy to 55 Gy, 5x/week, n = 64), both without additional boost. The equivalent dose of 2-Gy fractions (EQD2) was 55 Gy and 62 Gy, respectively. Late toxicity, assessed according to the LENT-SOMA criteria, and cosmetic outcome, graded on a 5-point scale, were evaluated after a median of 86 months (range 72-94 months) in tumor-free breast cancer patients. RESULTS: LENT SOMA grade 2/3 toxicity (2.5 Gy vs. 2.0 Gy): breast pain (18% vs. 11%; p = 0.3), fibrosis (57% vs. 16%; p < 0.001), telangiectasia (22% vs. 3%; p = 0.002), atrophy (31% vs. 3%; p < 0.001). Medication to breast pain was taken by 8% versus 9% of patients. Cosmesis was very good/good/acceptable in 75% versus 93% (2.5 Gy vs. 2.0 Gy; p = 0.006). CONCLUSION: Late morbidity was significantly frequent and cosmesis was significantly worse after hypofractionated radiotherapy (2.5 Gy to 55 Gy). However, morbidity was not associated with major implications on daily life. PMID- 16220401 TI - Organ-sparing treatment in muscle-invasive bladder cancer. AB - BACKGROUND AND PURPOSE: Organ-sparing treatment of bladder cancer by a trimodality approach is feasible and effective. In this study, the results of a series of patients are reported, who were, in the majority, not suitable for major surgery. PATIENTS AND METHODS: In the period from June 1995 through December 2003, 68 patients (64 males, four females) with urothelial bladder cancer were treated with curative intent. The median age was 68 years (range 42 82 years). Clinical T-category was 32x T2, 20x T3, and 16x T4. Transurethral resection was performed in all cases, and a complete TUR-BT (transurethral resection of bladder tumor) was attempted, if possible. Radiotherapy was administered in conventional fractionation (five fractions of 1.8 Gy per week) up to 50.4 Gy to bladder, and regional nodes and the whole bladder received a boost up to 54-59.4 Gy. 34 patients received concurrent cisplatin-based chemotherapy (25 mg/m(2) on days 1-5 and 29-33), and patients with impaired renal function were either treated with irradiation alone (n = 7) or received paclitaxel as alternative to cisplatin in a phase II protocol or on an individual decision (n = 27). The median follow-up was 34 months (range 2-104 months). RESULTS: A histologically confirmed complete remission (CR) on restaging cystoscopy was observed in 40/46 patients (87%) who underwent restaging cystoscopy. CR rates were not significantly correlated to T-category (CR: 24/32 T2, 9/19 T3, and 9/16 T4 tumors) or clinical nodal status. Patients with non-radical resection and macroscopic residual tumor (R2 resection) achieved a CR in only 39% (12/31); this figure was significantly lower as compared to patients with radical R0 TUR-BT (CR: 15/16, 94%, p = 0.013) Furthermore, age and preexisting anemia had no impact on response. The overall survival of the whole group was 45% after 5 years, and survival according to clinical T-category was 62% for T2, 43% for T3, and 19% for T4 (p = 0.015). In eleven patients, local disease progression or relapse was observed. So far, only one salvage cystectomy has been performed, due to contraindications to surgery in the majority of patients. CONCLUSION: The data obtained in this study confirm the high efficacy of TUR and radiochemotherapy for locally advanced bladder cancer. PMID- 16220402 TI - Fat necrosis after conserving surgery and interstitial brachytherapy and/or external-beam irradiation in women with breast cancer. AB - PURPOSE: To investigate the incidence of fat necrosis, breast tissue fibrosis and breast pain after conserving surgery and accelerated partial-breast irradiation (APBI, group A), whole-breast external-beam irradiation (EBRT, group B), or EBRT combined with an interstitial boost (EBIB, group C) in women with breast cancer. PATIENTS AND METHODS: 85 patients who received breast-conserving therapy from 02/2000 to 03/2002 were analyzed. 30 patients received EBRT, 33 patients APBI, and 22 patients EBIB. Median follow-up was 35.5, 35.0, and 37.5 months, respectively. Fat necrosis was detected and rated by mammograms, fibrosis and pain were clinically rated with the LENT-SOMA scores. RESULTS: The incidence of fat necrosis was 15.3% for all patients; and 15.2%, 20.0%, and 9.0% for groups A, B, and C, respectively. The 3-year fat necrosis-free survival probability was 83%, 76%, and 95% (difference not significant). The mean time to first diagnosis of fat necrosis was 25.6, 26.2, and 26.0 months. No patient needed surgical intervention because of fat necrosis-related pain. Fibrosis was present in 27.3%, 63.3%, and 77.3% (p < 0.001), breast pain in 9.0%, 33.4%, and 18.1% (p < 0.05). CONCLUSION: Asymptomatic fat necrosis is a common finding after both percutaneous and interstitial irradiation after breast-conserving surgery. This analysis did not support the hypothesis that APBI with multicatheter implants leads to higher rates of fat necrosis, fibrosis, or pain. PMID- 16220403 TI - Significant negative impact of adjuvant chemotherapy on health-related quality of life (HR-QoL) in women with breast cancer treated by conserving surgery and postoperative 3-D radiotherapy. A prospective measurement. AB - PURPOSE: To prospectively assess health-related quality of life (HR-QoL) in women after conserving surgery for breast cancer during/after postoperative 3-D radiotherapy. PATIENTS AND METHODS: 109 consecutively treated patients were analyzed. HR-QoL was assessed at initiation (t1), end (t2), and 6 weeks after radiotherapy (t3) using the EORTC modules QLQ-C30/BR23. Patients were divided into three therapy groups. Group I comprised 41 patients (radiotherapy and adjuvant chemotherapy), group II 45 patients (radiotherapy and adjuvant hormonal therapy), and group III 23 patients (radiotherapy alone). Reliability was tested. Scale means were calculated. Univariate (ANOVA) and multivariate (MANCOVA) analyses were performed. RESULTS: Reliability testing revealed mean Cronbach's alpha> 0.70 at all measurement points. ANOVA/MANCOVA statistics revealed significantly better HR-QoL for patients in group II versus I. Patients receiving radiotherapy alone (group III) showed the best results in HR-QoL. However, scale mean differences between groups II and III were not significant. CONCLUSION: HR QoL measurement using EORTC instruments during/after radiotherapy is reliable. Adjuvant chemotherapy significantly lowered HR-QoL versus hormones or radiotherapy alone. Chemotherapy patients did not recover longitudinally (from t1 to t3). PMID- 16220404 TI - Toxicity and survival results of a phase II study investigating the role of postoperative chemo-radioimmunotherapy for gastric adenocarcinoma. AB - BACKGROUND AND PURPOSE: To investigate the role of postoperative concomitant chemo-radioimmunotherapy in gastric adenocarcinoma patients. PATIENTS AND METHODS: 59 patients, who underwent total or subtotal gastrectomy, with lymph node involvement, positive microscopic surgical margins or serosal involvement were included in the study. Radiotherapy started concomitantly with chemotherapy and levamisole. Extended-field radiotherapy was given to gastric bed and regional lymphatics via two anterior-posterior/posterior-anterior fields. A total dose of 45 Gy in 25 fractions with a fraction size of 1.8 Gy was planned. In 28 patients (48%) with positive surgical margins a 10-Gy boost dose was given to the anastomosis site. An adjuvant i.v. bolus of 450 mg/m(2)/day 5-fluorouracil (5-FU) was administered concomitantly during the first 3 days and at the 20th day of irradiation. After completion of radiotherapy, i.v. boluses of 450 mg/m(2)/day 5 FU and 25 mg/m(2)/day rescuvorin were continued for 6 months once a week. Levamisole 40 mg/day orally was started at the 1st day of radiotherapy and also continued for 6 months. Median follow-up was 37 months (7-112 months). RESULTS: Median survival was 23 months. Overall 3- and 5-year survival rates amounted to 35% and 14%, respectively. Median survival of the patients with positive surgical margins was 22 months. The 3- and 5-year locoregional control rates were 59% and 55%, respectively. The most common toxicity was upper gastrointestinal system toxicity, which was observed in 42 patients (71%). Four patients (7%) died on account of early toxic effects, and six (10%) could not complete treatment. CONCLUSION: Although 48% of the study population involved patients with microscopic residual disease, the survival results as a whole were satisfactory. However, due to high toxicity, radiotherapy must be delivered with the most proper techniques along with adequate nutrition and supportive care. PMID- 16220405 TI - Lymphatic fistulas: obliteration by low-dose radiotherapy. AB - BACKGROUND: Lymphatic drainage from the surgical wound is an uncommon but challenging complication of surgical intervention. Protracted lymphorrhea contributes to morbidity, favors infections and results in a prolonged hospital stay. Treatment options include surgical ligation and, more conservatively, leg elevation, continuous local pressure, subatmospheric pressure dressings, and low dose radiotherapy. This study examines the efficacy of low-dose radiotherapy. PATIENTS AND METHODS: 17 patients (19 fistulas) with lymphorrhea following vena saphena harvesting (n = 7), femoropopliteal bypass (n = 3), varicose vein surgery (n = 2), hip arthroplasty (n = 3; five fistulas), shunt surgery (n = 1), and piercing (n = 1) were referred for external radiotherapy. Depending on the depth of the fistula, orthovoltage (n = 12), electrons (4-11 MeV; n = 2) or photons (8 MV; n = 3) were used. Fractions between 0.3 Gy and 2 Gy were applied; the individual total dose depended on the success of the radiotherapy, i. e., the obliteration of the lymph fistula, and varied from 1 to 12 Gy. RESULTS: In 13 out of 17 patients complete obliteration of the fistula was achieved. Interestingly, this was achieved in nine of the ten patients irradiated with total doses of 0.05), postprandial gallbladder volume (PGV) was lower in IBS group than in the control group (5.5 +/- 1.4 ml vs. 6.2 +/- 1.9 ml, p = 0.03). Accordingly, the mean EF of gallbladders was found to be higher in IBS group than in the control group (69.1% vs. 64.1%, p = 0.0001). There were no difference in FGV, PGV and EF of gallbladders between constipation predominant and diarrhoea predominant IBS patients (p >0.05). CONCLUSION: The results of this study indicated that IBS patients have increased emptying of gallbladder compared to healthy subjects. PMID- 16220413 TI - [Psychological trauma in psychiatric textbooks published in german - the dominating view of psychiatrists between 1945 and 2002]. AB - AIM: The study investigates the concepts of psychological trauma and their changes over time in psychiatric textbooks published in German between 1945 and 2002, assuming that textbooks reflect the established and dominating views of their time. METHOD: [corrected] In psychiatric textbooks, the terminology, concepts of illness, and recommendations for assessment and treatment concerning psychological trauma were analysed. RESULTS: The concept of psychological trauma that had existed since 1916 continued to dominate textbooks up until the 1960s. Findings on holocaust survivors entered textbooks not before the mid 1970s. Since the mid 1990s, the concept of Post-Traumatic Stress Disorder has been widely established in textbooks. CONCLUSION: Changes of dominating views on this issue in textbooks appear to have been extremely slow and occurred with significant delays in the past. The change of the dominating view in the 1970s was linked to the establishment of a new generation of leading psychiatrists. Since the introduction of PTSD, psychiatric textbooks have given up a previously negative attitude towards patients suffering from psychological trauma. PMID- 16220414 TI - [Psychiatric day hospitals -- quo vadis?]. PMID- 16220415 TI - [For and against: controlled drinking as useful and necessary treatment alternative]. PMID- 16220416 TI - [Post-traumatic stress disorder: reserve is necessary]. PMID- 16220417 TI - [The "war neurosis"-- an early model of a pluridimensional outlined trauma-theory in psychiatry]. AB - The war neurosis, mainly observed during the first World War, caused a lively debate on its origin and etiology. In psychiatric history this debate is often portrayed in a somewhat simplifying manner. Namely Oppenheim was considered as the leading exponent of the organic etiology of traumatic neurosis, while others, with growing interest in psychodynamic theories, were thought to favor exclusively psychogenesis of this condition. However, only discussing matters in this way would be too simple. Rather, the controversy was much more differentiated and led to important insights: First, it turned out to be impossible to explain any psychopathological syndrome exclusively in terms of neuropathological, i. e. structural alterations, in particular, a syndrome presenting with a sudden onset of dissociative and conversion symptoms. Secondly, the psychiatric theory of hysteria of Charcot and Freud was developed and extended further. And, last, the etiology of war neurosis was recognized to be multifactorial. Thus, the discourse on this issue was not one-dimensional and favoring a single explanation for a complex disorder, but revealed "pluridimensional" features. Subsequently psychotherapy, for the first time, was widely introduced into clinical practice with a clear indication and well-defined methodological approach. In summary, war neuroses can be addressed as an early model of traumatic stress disorder, such as acute stress disorder or post traumatic stress disorder. PMID- 16220418 TI - [Patient evaluation of day clinic treatment -- experience of treatment and change]. AB - At the end of their treatment, 48 patients, having attended the day clinic of a general psychiatric hospital, evaluated important aspects of their therapy. These evaluations were analysed. In resulting with the norm sample of the inpatient experience questionnaire (SEB), more positive values were obtained for all the scales. The scales "experiences of relationship to individual therapist" and "experiences of relationship to the therapeutic team" yielded the highest values for patient satisfaction. We found that the categories of "general documentation about change through treatment" within the Psy-BaDo are relevant for patients attending a day clinic treatment. There is evidence for a large improvement with respect to all categories. Highest changes were reported for the subjects "understanding of the individual illness", "body symptoms" and "psychological well-being". Smaller changes were reported for the subjects "private relationships", "relationships in job" and "social problems". Compared to the patients, the therapists' estimate of the changes were lower. On an individual basis, the assessments of the patient and therapist agreed well. Sociodemographic characteristics and other criteria of the illness and therapy were not well associated with the scales of the SEB and the categories of Psy-BaDo. Only the duration of symptoms and the frequency of previous inpatient therapies were clearly associated with the categories of Psy-BaDo. The data, derived from standardized questionnaires, enable comparisons to be made between different institutions and provide evidence for a very positive evaluation of the investigated treatment. PMID- 16220419 TI - [Compliance in the treatment of schizophrenia -- results of an expert survey in Saxony and Saxony-Anhalt]. AB - OBJECTIVE: To report attitudes of psychiatrists and neurologists towards compliance with regard to medication-taking behaviour of patients suffering from schizophrenia. METHOD: A questionnaire was sent to psychiatrists and neurologists practising in Saxony and Saxony-Anhalt. Out of a total of 120, 54 completed questionnaires were returned. RESULTS: Most of the experts defined compliance as the degree of fidelity to treatment recommendations and patient cooperation. In the experts' view, a positive attitude towards treatment, insight into illness, and accurate perception of the symptoms correlate strongly with compliance, whereas the association between sociodemographic characteristics and compliance was seen as moderate. A majority of experts considered there were changes in compliance over time. DISCUSSION: With regard to the improvement of compliance, the importance of the doctor-patient-relationship is supported by the results of our study. As ratings of doctors suggest, compliance is a broadly determined behaviour which is variable over time. Future research on medication-related compliance in patients suffering from schizophrenia should take into account these findings. PMID- 16220420 TI - [Prevention -- a topic in social-psychiatric research?]. AB - OBJECTIVE: Currently, substantial effort is made to establish prevention as one of the main parts of the German health care system. In this context we investigated the current role of prevention in social-psychiatric research. METHODS: We analysed original papers published in the journal Psychiatrische Praxis in the years 2003-2004 by computer-aided full text analysis. RESULTS: 16.5 % of the studies mentioned prevention, in one fourth of these, prevention was the main topic. Subject of these papers were suicide prevention in psychiatric patients, relapse prevention in mentally ill offenders, and prevention of acute confusional state after hip surgery. Most of the work was dedicated to tertiary prevention, followed by secondary prevention. Two papers mentioned primary preventive aspects and only one report was dedicated to primary prevention as a main topic. CONCLUSIONS: Although early diagnosis and intervention as well as rehabilitation of mental disorders are main topics of social psychiatry, only few reports relate explicitly to prevention. The role of social psychiatry in primary prevention has to be defined. PMID- 16220422 TI - [The serotonin syndrome]. AB - The serotonin-syndrome is a possible side-effect in the treatment with serotonergic drugs. There are diagnostic criteria for diagnosis of this syndrome. After discontinuation of administering the serotonergic drug is fully reversible. PMID- 16220421 TI - [Amisulpride in Huntington's disease]. AB - OBJECTIVES: Amisulpride is a substituted benzamide derivative with atypical antipsychotic properties and low side effects. METHOD: We report four cases of patients with clinically and genetically established Huntington's disease and signs of psychosis who were treated with Amisulpride. RESULTS: Two patients developed extrapyramidal side effects due to the treatment. The antipsychotic therapy of all patients was effective. DISCUSSION: Due to degeneration of striatal neurons patients suffering from Huntington's disease react early with development of extrapyramidal side effects after therapy with amisulpride. PMID- 16220425 TI - [A newspaper analysis of psychiatry in 1913]. PMID- 16220428 TI - [Fractures of the accetabulum--treatment strategies and actual diagnostics]. PMID- 16220431 TI - [Quality assurance in operative medicine--a part of the clinical research]. PMID- 16220432 TI - [Options for quality improvement in the treatment of rectum carcinoma]. AB - By means of a prospective multi centre study, 13 419 cases of surgically treated patients with rectum carcinomas were registered between 1.1.2000 and 31.12.2003 and assessed in regard to possible problems concerning indications and operative procedures. Beside a high rate of non-local resective procedures in T1-low risk carcinomas, unnecessary extirpations in cases of tumour localisation over 8 cm from the anal verge were found. Tumours of the lower two-thirds of the rectum were treated by incomplete TME in 20 % of the patients. In addition, there seems to be too low a rate of neo-adjuvant therapy procedures. Protective stomata were frequently foregone after low anterior resection. Endoscopic interventional methods were still used reluctantly in inoperable situations. PMID- 16220433 TI - [Quality of life and sexuality after surgery for rectal cancer--a follow-up study]. AB - BACKGROUND: Previous studies have suggested that sexuality following surgery for rectal cancer is better in women than in men and differs with type of surgery in terms of impact on Quality of Life. However, the findings were inconsistent. The aim of the presented study was to address these issues. METHODS: 570 patients that had undergone surgery for rectal cancer in our department from 1992 to 1999 were included. After determination of survival status questionnaires on Quality of Life were sent to 370 patients one to two years post surgery. We analysed data from 215 patients of which 103 were female and 112 male. 248 patients died during the time observed. Statistical analysis was done with descriptive methods, Kaplan Meier analysis (log rank test), T-test and analysis of variance using SPSS 11.0 for Windows. RESULTS: Significant differences were seen in symptom scales between men and women. Women scored higher for distress through medical treatment and had higher values for insomnia, fatigue and constipation. Both sexes had impaired sexual life but men had significantly higher values and felt more distressed by the impairment of sexuality than women. For patients receiving abdominoperineal resection sexuality was most impaired. Anterior resection with pouch had a detrimental effect on diarrhoea and treatment strain. CONCLUSION: The study showed that sexuality is influenced by gender and type of surgery and impacts Quality of Life after surgery for rectal cancer. Assessing quality of life with generic and specific instruments is helpful to determine differences between surgical procedures, age, gender, and adjuvant therapy where standard parameters such as survival have their limitations. PMID- 16220434 TI - [Functional results after transvaginal, transperineal and transrectal correction of a symptomatic rectocele]. AB - INTRODUCTION: The aim of this study was to compare functional outcome after transvaginal, transperineal and transrectal repair of a symptomatic rectocele and to develop the ideal surgical approach. PATIENTS AND METHOD: 28 patients (27 female, 1 male) who had undergone rectocele repair from 1996 to 2003 were analysed. Mean age was 59 years (range 30-79 years), follow-up was 24 months (range 3 to 70 months) and mean appearance of symptoms was 4 years prior to the operation (6 months-32 years). Transvaginal repair was performed in 13 cases, transperineal repair in 8 cases and transrectal repair in 7 cases. RESULTS: 24 of 28 patients (85.7 %) are satisfied with the operation-result (transvaginal 12 of 13 patients [92.3 %], transperineal 7 of 8 patients [87.5 %] and transrectal 5 of 7 patients [71.4 %]). 25 patients (89.3 %) are free of complaints or describe an evident improvement of symptoms (transvaginal 12 of 13 patients [92.3 %], transperineal 7 of 8 patients [87.5 %] und transrectal 6 of 7 patients [85.7 %]). There is one postoperative dyspareunia. DISCUSSION: Best treatment of a rectocele starts with patients selection. Considering pelvic floor as functional unity, concomitant urologic-gynaecologic lesions and proximal intraabdominal disturbances the appropriate surgical procedure is selected. CONCLUSION: Surgical approach to correct a symptomatic rectocele depends on the concomitant lesion. PMID- 16220435 TI - [Impact of pneumoperitoneum on expression of E-cadherin, CD44v6 and CD54 (ICAM-1) on HT-29 colon-carcinoma cells]. AB - INTRODUCTION: The mechanism of potential tumor cell spread and growth during laparoscopy is poorly understood. Nevertheless, different experimental studies reported a stimulation of tumor cell growth and an increased metastatic potential of carcinoma cells using carbon dioxide as an insufflation medium. Adhesion molecules do play an important and regulatory function in the process of metastatic spread and invasion of cancer cells. Therefore we investigated the influence of CO2 and Helium insufflation on the in-vitro expression of E Cadherin, CD44v6 and CD54 (ICAM-1) on HT-29 colon carcinoma cells. METHODS: HT-29 carcinoma cells were exposed to either CO2 or helium insufflation. Expression of E-Cadherin, CD44v6 and CD54 (ICAM-1) on HT-29 colon carcinoma cells were measured 1, 12, 24, 48 and 96 h after CO2 and helium insufflation using flowcytometry (FACScan). Data were analyzed by Friedman-test. RESULTS: HT-29 cell line showed a short decrease in E-Cadherin expression after CO2 exposure while helium insufflation had no influence. In contrasts to these findings the expression of CD44v6 and CD54 on HT-29 cells were not influenced significantly by either CO2 or helium. CONCLUSION: CO2 seems only to have a minor influence on the expression of E-Cadherin while expression of other adhesion molecules did not change after CO2 incubation. The alternative gas helium did not cause any significant changes of the expression of either E-Cadherin, CD44v6 and CD54. Further investigations are needed to elucidate the changes of the metastatic potential of tumor cells after laparoscopic and open procedures. PMID- 16220436 TI - [Long-term results after gastric banding]. AB - BACKGROUND: Adjustable gastric banding is a popular bariatric operation in Europe. The rate of long-term complications like pouch dilatation, slippage and band migration and the long-term effect of weight loss are reported in meta analysis and few studies for a period of more than five years. We report on experiences after gastric banding. METHODS: Over a period of 10 years 168 patients with morbid obesity were treated with gastric banding. Preoperative data, postoperative weight loss and long-term complications were prospectively obtained and retrospectively analyzed. RESULTS: Mean age of the patients was 41.7 years with a mean preoperative BMI of 49.6 kg/m2. No intraoperative or postoperative death occurred in the first 30 postoperative days. Intraoperative conversion rate was 7.1 %. 79.8 % of the patients (n = 134) were available for follow up (mean follow-up time 66.7 months). Long-term complications occurred in 22.5 % of the patients. 30 complications (17.8 %) were related to the band and 8 (4.7 %) to the access-port or to the tube. Mean excess weight loss was 39.6, 47.3, 44.2, 43.4 and 32 % after 1, 2, 4, 5 and 8 years. CONCLUSIONS: Laparoscopic gastric banding can achieve an effective weight loss. However band-related and functional complications will influence the late outcome. Pathways to choose the best surgical method for the individual patient are necessary to reduce failures after gastric banding. PMID- 16220437 TI - [Study of quality assurance "surgical treatment of morbid obesity" since 1.1.2005]. AB - Since January 1st 2005, the situation of bariatric surgery has been examined in Germany. The data are registered in cooperation with the An-institute of quality control in surgery at the Otto-von-Guericke-Universitat Magdeburg. The data registration occurs in an internet on-line data bank. Application for participation in this study is available on our correspondence address. All hospitals carrying out bariatric surgery are asked to take part in this study. PMID- 16220438 TI - [The thoracoscopic thymectomy for myasthenia gravis]. AB - Myasthenia gravis (MG) is a chronic autoimmune disease that usually responds positively to treatment with thymectomy. Standard surgical procedures have been shown to result in a favorite outcome. The optimal surgical access, however, is under discussion. PATIENTS AND METHOD: In a prospective study conducted between 8/97 and 4/05, 173 patients with MG underwent thoracoscopic thymectomy, for which a left-sided approach was generally applied. An analysis of the intraoperative and postoperative course was performed in 137 patients (8/97 and 12/03), as well as of the impact of the surgical procedure on further development of the disease. The results obtained were compared with those published in the literature, with particular reference to results obtained with open surgery. In 8 patients, the procedure was converted to an anterolateral thoracotomy or sternotomy, so we examined 129 patients, which were operated thoracoscopically. RESULTS: The patients in this study were 93 females and 36 males, with a mean age of 35.8 (range: 9-83) years. The mean preoperative duration of the disease was 22.9 (range: 1-140) months. The duration of operation was 50 to 85 minutes, the blood loss was lower than 100 ml. The mean follow up was 24.7 (range: 1-57) months. Complete remission was noted in 30 patients (23.3 %), and improvements reflected either in a reduction of the required medication, or a decrease in symptom severity, were seen in a total of 86 patients (66.6 %). CONCLUSION: Complete thoracoscopic thymectomy is a technically feasible operation, and as effective as conventional open surgery. Remissions or symptomatic improvements were observed in more than 89 % of the patients. The low morbidity rate, in combination with excellent cosmetic results, has led to increasing acceptance of the operation both by patients and neurologists. Therefore, thoracoscopic thymectomy represents a new, alternative method for treatment of patients with MG. PMID- 16220439 TI - [Thyroid surgery: generalist or specialist?]. AB - BACKGROUND: Thyroid surgery is done in Germany in a considerable numbers of operations (about 110,000 per year). To perform thyroid operations by so called "generalists" or "specialists" have been discussed intensively, however, this issue have not been analyzed in detail. METHODS: Study material comprised 16,500 consecutive thyroid operations with 30,000 operated sites that have been prospectively documented in the German Thyroid Multicenter Study performed 1998 through 2001. Quality of surgery were analyzed by calculating the inverse relationship between volume and outcome (complication rate). RESULTS: To achieve complication rates (permanent unilateral recurrent laryngeal nerve paralysis and hypocalcemia) of < 1 % (primary surgery) or < 3 % (redo surgery) the minimum number of thyroid operations of lower level of experience (e. g. benign nodular goiter) per year was n=30. The minimum number of operations with higher level of experience was significantly lower (n=3-12) due to the higher level of experience of operating surgeons. In contrast to the rates of postoperative hypocalcemia the rates of postoperative recurrent laryngeal nerve paralysis was clearly related to the number of thyroid operations performed. CONCLUSIONS: The high number of thyroid operations in Germany with about 20 % of operations of high level experience are requiring surgical curricula and hospital structures that offer as well generalists as specialists to treat the broad spectrum of thyroid diseases accordingly. To lower the complication rate especially of difficult thyroid operations the level of specialization in Germany have to be increased. PMID- 16220440 TI - [Decision making in postoperative incidentally found small C-cell-carcinoma]. AB - Surgical therapy of incidentally postoperative diagnosed small sporadic medullary thyroid cancer (MTC) is discussed controversially. In principle completion thyroidectomy with neck dissection and regulary tumor follow-up are under discussion. A total of 277 patients with MTC were treated between 1986 and 2004. In 22 cases diagnosis of a small (pT1 or pT2) sporadic MTC was incidental and only postoperatively confirmed. Normally total thyroidectomy with neck dissection is standard surgical therapy of a known MTC. Because of postoperative incidental diagnosis in all 22 cases surgical therapy was less then total thyroidectomy. Mutation analysis of RET Proto-Oncogen and familial history were negative in all cases. All patients were systematically followed-up in defined intervals by calcitonin, pentagastrin stimulation test, carcinoembryonic antigen and ultrasound. Median follow-up is 6.2 years (range: 2-13 years) and although a hemithyroidectomy or less was performed all 22 patients are cured by the MTC. We conclude that completion thyroidectomy and neck dissection are not mandatory in such patients, if the tumor is completely resected and genetic background is excluded. Indispensably a systematic long term follow-up of at least 10 years, better a life-long, is mandatory. PMID- 16220441 TI - [The farmost unknown disease of postoperative permanent hypoparathyroidism- feasibility and validation of an innovative questionnaire instrument]. AB - Aim of our study has been the systematic evaluation concerning the still farmost unknown disease of postoperative, permanent hypoparathyroidism. For this we designed a specific questionnaire as measuring instrument--it was evaluated by experts and validated by 3 control groups--to examine then a patient group (n = 20) with symptomatic postoperative, permanent hypoparathyroidism. The application of the questionnaire was very feasible, the reliability of measuring scales according to Cronbach's alpha > or = 0.70. The validity of content and construct validity fully existed (Pearson's correlation coefficient r > or = - 0.27; p < 0,05). The farmost frequent clinical complaints were peripheral cold sensations (65 %), pains in joints (55 %), sensation of heaviness and weakness in extremities (55 %) and paresthesias (45 %). Most frequent clinical symptoms were brittle nails (40 %) and diarrhoea (35 %). Sum and distress level scores for classification of illness severity were determined. In conclusion our questionnaire is a valid instrument and from a clinical point of view easy to apply, reasonable and very feasible in practical use. It represents a novel, optimum instrument to determine disease condition and illness severity of hypoparathyroid patients and makes it possible to gain further information of this up to now less systematically analysed disease. PMID- 16220442 TI - [Sporadical extraperitoneal desmoid tumors--review and report on 4 cases]. AB - INTRODUCTION: Desmoid tumors are very rare mesenchymal tumors with a partially aggressive growth pattern and high relapse rates. Without metastases, they are classified as semi-malignant tumours. The treatment of this neoplasia is discussed controversially. Up to now, there exists no broad consensus on the treatment regimes. Traditionally, the primary radical surgical resection is often preferred. This might be combined with an (neo-)adjuvant radiation therapy. Recently an increasing importance of radiotherapy has been described. Several studies favour radiotherapy as the single form of treatment. Furthermore several medication regimes have been described in the literature. CASE REPORTS: We report the follow-up of 4 patients who were treated for desmoid tumours. In 2 cases surgery was the primary treatment. After R0-resection and adjuvant radiation therapy, one of the patients developed 7 relapses. In another patient, 5 relapses were observed after surgical therapy. DISCUSSION: Therapeutic guidelines with broad acceptance do not exist up to now due to missing data of prospective studies with sufficient case numbers. In our opinion wide surgical resection should be performed in primary occurring cases with resectable tumours. On account of the previous experiences, it can be concluded that therapy of primary non respectable or recurrent desmoid tumours is not only a domain of a surgical approach. At present an adequate treatment should be planned interdisciplinary by surgeons, radiation therapists and oncologists. For generally accepted guidelines for this rare disease, general therapeutic options based on prospectively randomised studies have to be elaborated. PMID- 16220443 TI - [Clinical and functional results after transfemoral thrombectomy for iliofemoral deep venous thrombosis: a 5-year-follow-up]. AB - PURPOSE: The optimal therapy of deep pelvic and leg venous thrombosis is still a matter of debate. The purpose of our study was to evaluate early and late results of iliofemoral thrombectomy with regard to the prevention of the development of a postthrombotic syndrome. METHODS: Between 1996 and 2000, 57 patients underwent transfemoral venous thrombectomy for acute iliofemoral thrombosis. 30 patients were reexamined after a mean of 60.4 months. At follow-up, the patency of the venous segments as well as the development of reflux was investigated by duplex ultrasound. Furthermore, clinical signs and symptoms of chronic venous insufficiency as well as the subjective satisfaction of the patients with the operation were recorded. RESULTS: Postoperatively the veins of the lower leg were completely recanalized in 25 % of the cases, those of the thigh in 52.3 %. The patency rate at the level of the groin, the pelvis and the caval vein were 92.5, 86 and 100 %, respectively. At follow-up, the veins of the lower limb, the thigh and the pelvis were patent in 76.7 % each and in 73.3 % at groin-level. The caval vein was completely recanalized in all cases. Reflux occurred in 12 patients. 26.7 % of the patients showed no signs of a postthrombotic syndrome. 63.3 % had mild changes including dilated superficial veins and swelling tendency, and only in 10 % trophic skin changes were apparent. A healed or active ulceration did not occur in any case. Except one, all patients were satisfied with the results of the thrombectomy. CONCLUSIONS: Transfemoral thrombectomy for acute iliofemoral venous thrombosis offers good early and late results in terms of preserving venous function and reducing symptoms of venous insufficiency. In our patient population, the development of a severe postthrombotic syndrome could be reduced effectively. PMID- 16220444 TI - [The physics of vacuum therapy]. AB - BACKGROUND: Vacuum therapy is a routine and successful method for wound treatment and for the temporary covering of soft-tissue defects. It is an occlusive method that can be used for treating acute, chronic and infected wounds. To date, no data are available regarding secretion transit times and pressure conditions in lesions treated with vacuum therapy. The present study had as its objectives to examine the mechanisms of vacuum therapy and determine the effects of physical forces on the wound surface with the purpose of formulating recommendations for pressure settings using the various available vacuum pumps. MATERIAL AND METHODS: Using an appropriate model, we measured secretion transit times and pressures using two different vacuum therapy pads. We then conducted pressure measurements during dressing change in eight patients with wound surfaces greater than 20 cm(2). RESULTS: The secretion transit times remained unchanged with the polyvinyl alcohol (PVA) pad and a negative pressure of 40 kPa, but decreased by about 50 % when the black polyurethane (PU) pad was used. Pressure measurements showed that, at this negative pressure, there was only a slight positive external pressure of 31 mmHg on the wound surface. CONCLUSION: When the PVA pad is used, negative pressures greater than 40 kPa should be applied in order to effect a maximum transit of wound secretion and prolongation of the use of the vacuum dressing. When the PU pad is used, the applied negative pressure can be lower due to the pad's larger pores, as recommended by Argenta and Morykwas. An excessive external pressure on the wound surface does not occur. PMID- 16220445 TI - [Bone mineral density in postmenopausal women with proximal femoral fractures- comparative study between quantitative ultrasonometry and gold standard DXA]. AB - INTRODUCTION: Osteoporosis associated proximal femoral fracture is a major public health problem. Diagnostic assessment includes patients history, laboratory testings and bone mineral density measurements. Hereby, dual X-ray absorptiometry (DXA) is regarded as the "Goldstandard". Quantitative ultrasonometry (QUS) of bone is a safe, simple, free of radiation, portable, cost-effective and therefore powerful diagnostic tool. QUS should be taken in account for primary assessment in patients with supposed osteoporosis in clinical practice. AIM OF THE STUDY: We performed this cross sectional pilot study to evaluate the ability of two different QUS-devices (os calcis) in comparison to DXA (lumbar spine and femoral neck) to discriminate postmenopausal women with proximal femoral fractures from healthy, age- and BMI-matched controls. PATIENTS AND METHODS: All together, 44 postmenopausal women were included. Of these, 22 suffered a proximal femoral fracture and were compared with 22 healthy, age- and BMI-matched controls. Bone assessments were performed by DXA (femoral neck and lumbar spine) and QUS of the heel using Achilles and Insight. RESULTS: T- and Z-Score of DXA (femoral neck) were significantly lower in women with hip fracture compared to controls (p < 0.008 and p < 0.01). QUS-Insight also revealed significantly lower values of T- and Z-Score in women with hip fracture compared to controls (p < 0.01 and p < 0.005). QUS-Achilles measurement results also comprised significant differences between the groups (T-Score und Z-Score: p < 0.02). In accordance to the T-Score (femoral neck), all three devices (DXA femoral neck, Achilles and Insight) showed an equal significant correlation (p < 0.001). The correlation in between both QUS devices was higher (0.956; p < 0.0001) than in between DXA-results (femoral neck vs. lumbar spine, 0.577; p < 0.01). The Z-Score also showed a significant correlation. DXA (lumbar spine) didn't show any significant differences in T- and Z-Score. CONCLUSION: In comparison to the gold standard DXA (femoral neck), both QUS-devices showed an equal ability to significantly discriminate postmenopausal women with proximal femoral fractures from healthy, age- and BMI-matched controls. If our results are confirmed by more cross-sectional and longitudinal studies, QUS could be a helpful and valuable technique in clinical practice. Finally, all patients with osteoporosis-related fractures should be thoroughly investigated during their hospitalisation and effective treatment must be instituted. PMID- 16220446 TI - [Vertebroplasty--basic science, indications and technique]. AB - INTRODUCTION: Vertebroplasty is a relatively new procedure for the treatment of vertebral body fractures of different origin. Due to early promising results the rate of performed procedures increases constantly. Indeed an increasing number of partially severe complications are reported. A summary of the existing findings seems therefore necessary. METHODS: On the basis of a medline inquiry important aspects of vertebroplasty in the fields of basic science, diagnostics, indications, contraindications, technique, results and complications were compiled. RESULTS: Especially for the field of indications, possible longterm effects and for the "ideal" technique open questions exist, mainly due to missing prospective, randomized long-term clinical trials. CONCLUSIONS: A final assessment of the significance of vertebroplasty for osteoporotic fractures in comparison to conservative treatment is actually not possible. In contrast vertebroplasty is a therapeutical option for malignomas in selected cases. PMID- 16220448 TI - [Three Jewish Germans as chief surgeons in the Berlin-Friedrichshain Hospital--a historical reminiscence]. AB - Three jewish Germans were between 1903 and 1933 continuously in charge of a department of surgery at the oldest municipal hospital in Berlin-Friedrichshain (founded 1874). They were A. Neumann (1865-1920), Moritz Katzenstein (1872-1932) and Max Marcus (1892-1983). A. Neumann described in 1907 the so-called omentum majus-cuff. By this a drain is wrapped up and put into a perforated ulcus pylori, which cannot be treated by another procedure. M. Katzenstein in 1900 was the first in Germany who repaired a torn off meniscus by suture with a good result (The very first was T. Annandale in Edinburgh in 1883). Aside of this procedure, important for prevention of gonarthrosis, M. K. described new methods for repairing torn ligaments of the elbow and the ankle joint. He was a close friend of Albert Einstein during his years in Berlin. Max Marcus was after the death of M. Katzenstein the youngest chief surgeon in Berlin--but only until April 1933. Then he was expelled by the Nazis, because he was a jew. In June 1933 he immigrated to Palestine and became the leading surgeon there in the later Israel. After the end of World War II he refused coming back to Germany, though F. Sauerbruch spoke about him as the "great hope for the German surgery". PMID- 16220447 TI - [Long-term results in the treatment of foot injuries in polytraumatized patients]. AB - INTRODUCTION: The aim of this study was to evaluate the long-term outcome of multiple injured patients with foot injuries. While severity and frequency of multiple injuries could be decreased in the last years, it was not possible to decrease the severity of injuries of the foot and ankle region. These injuries are often not detected and their severity is underestimated in primarily diagnostics what might lead to complaints in the longterm course. METHODS: The multiple trauma database of the Department of Traumatology of the University of Vienna includes 386 patients from September 1992 to April 2001. 40 (10.4 %) of these patients suffered a fracture or dislocation of foot or ankle. 33 (82.5 %) multiple injured patients with injuries of foot or ankle could be reexamined between April and September 2002 and the longterm outcome of the foot and ankle region could be evaluated objectively and subjectively. RESULTS: 22 (66.7 %) patients were male, 11 (33.3 %) female. The mean age was 34.5 years. 27 (81.8 %) patients still suffered from complaints related to their injured foot and ankle region. After calculation of the AOFAS these complaints were predominantly located in the ankle-hindfoot region. 22 (66.7 %) patients had to limit their sports activity. DISCUSSION: In our study period an increasing number and severity of injuries of foot or ankle can be shown. Furthermore multiple injured patients with concomitant injuries of foot or ankle show a lower ISS what suggests that these patients might have a higher survival rate and consecutively enter rehabilitation process. PMID- 16220449 TI - [Mechanisms of disease]. PMID- 16220450 TI - [Comparison of 3 types of covered self-expanding metal stents for the palliation of malignant dysphagia: results from the prospective Ludwigshafen Esophageal Cancer Register]. AB - BACKGROUND: Endoscopic stenting with self-expanding metal stents (SEMS) has proved to be an efficient palliative endoscopic therapy option in treating malignant dysphagia. Different types of SEMS have been examined previously in small randomized clinical trials. METHODS: A total of 150 consecutive patients (from 1/1995-1/2004; 35 females, 115 males) with a median age of 66 years (range 43-94 years) were evaluated. Three different types of SEMS were examined: Group A: Gianturco Z Stent n = 59 pts.; Gruppe B: Ultraflex Stent n = 49 pts.; Gruppe C: Flamingo Wallstent n = 42 pts.). Dysphagia was categorized into grade 0 (no dysphagia) to grade 4 (total obstruction). RESULTS: Stent insertion was successfully evaluated in 55/59 (93 %) of the patients in group A, in 47/49 (96 %) of the patients in group B and in 40/42 (95 %) of the patients in group C (p = 0.78). After stent insertion, a reduction in the dysphagia score from 2.77 to 0.99 was observed in group A, from 2.58 to 0.72 in group B and from 2.62 to 0.81 in group C (p = 0.37). The median time to dysphagia recurrence was 78 days in group A, 90 days in group B and 70 days in group C (p = 0.45). The median overall survival time was 96 days in group A, 118 days in group B and 108 days in group C (p = 0.18). The incidence of high grade complications was 34 % in group A, 16 % in group B and 17 % in group C (p = 0.04). CONCLUSIONS: The 3 types of SEMS examined in this study showed similar efficacies in treating malignant dysphagia. However, the major complication rate of the Gianturco Z stent was significantly higher when compared to the complication rate of the Ultraflex stent and the Flamingo Wallstent. PMID- 16220451 TI - Frequency of colorectal polyps in patients with sporadic adenomas or adenocarcinomas of the papilla of vater--an age-matched, controlled study. AB - BACKGROUND: Epithelial tumors of the papilla of Vater are rare neoplasms of the gastrointestinal tract. The carcinogenesis of these tumors seems to be fairly analogous to the genetic mechanisms which have been described for colorectal carcinoma. Patients with familial adenomatous polyposis bear a particularly increased risk for periampullary tumors. Data on whether the prevalence of colorectal tumors is increased in patients with sporadic ampullary neoplasms are scarce. METHODS: 26 consecutive patients (16 women, 10 men; median age 59 years) with sporadic adenomas (n = 19) or adenocarcinomas (n = 7) of the ampulla of Vater were retrospectively evaluated. The study patients were compared with 104 age-matched asymptomatic controls. All patients had undergone total colonoscopy. RESULTS: Neoplastic colorectal polyps were present in a similar proportion (23%) of patients of the study group compared with 26% in the control group (p > 0.05). Overall, 16 polyps were found among patients with ampullary tumors and 40 in asymptomatic controls (p > 0.05). Colonoscopy detected rectal carcinoma in 2 patients (8%) of the study group. Patients with and without colorectal polyps differed neither significantly by age nor by ampullary histological findings. 50% of the colonic polyps in patients with ampullary neoplasms were located in the ascending colon. CONCLUSIONS: The frequency of colorectal polyps in patients with ampullary tumors did not exceed the risk in the control group. However, the finding of 2 rectal carcinomas among patients with ampullary neoplasms supports the place of screening colonoscopy for the diagnostic work-up of ampullary tumors. Prospective multicenter studies should address this issue to provide a broad basis for future recommendations. PMID- 16220452 TI - Severe acute cholestatic hepatitis by infiltration of monoclonal plasma cells in multiple myeloma. AB - BACKGROUND: Plasma cell infiltration of the liver can be detected in 25 to 40% of patients with multiple myeloma. However, there are only rare cases of multiple myeloma clinically presenting as acute liver disease. CASE REPORT: We report an 88-year-old woman with painless jaundice and abnormal liver function tests, resembling acute cholestatic hepatitis. Viral hepatitis as well as autoimmune hepatitis could be excluded. Liver biopsy revealed a diffuse portal and sinusoidal infiltration of plasma cells with lambda light chain restriction. Serological immune fixation disclosed monoclonal gammopathy of IgG lambda with bone marrow infiltration of 25% plasma cells. After administration of 60 mg prednisolone per day, the elevated liver enzymes declined considerably. CONCLUSION: Hepatic plasma cell infiltration of multiple myeloma can, in rare cases, manifest as acute cholestatic hepatitis, which may respond to treatment with corticosteroids. PMID- 16220453 TI - Mechanisms of disease: PI3K/AKT signaling in gastrointestinal cancers. AB - The lipid kinase phosphoinositide 3-OH kinase (PI3K) and its downstream target Akt, also known as protein kinase B (PKB), are crucial effectors in oncogenic signaling induced by various receptor-tyrosine kinases. In recent years, data are accumulating that PI3K/Akt signaling components are frequently altered in a variety of human malignancies. This review summarizes the major effects of PI3K/Akt signaling on proliferation, survival and resistance to apoptosis, angiogenesis and cell motility in gastrointestinal cancers. In addition, activation of PI3K/Akt by various growth factors, the modulation of downstream targets by Akt-induced phosphorylation as well as novel treatment strategies targeting this pathway in gastrointestinal tumors are discussed. PMID- 16220454 TI - [Neoadjuvant therapy for resectable esophageal cancer]. AB - BACKGROUND: Surgical excision remains the only therapeutic approach with curative potential in patients with localized esophageal cancer. Due to the presence of lymph node metastases upon diagnosis in a large percentage of patients with locally advanced tumors and/or the presence of considerable co-morbidity, only a limited number of patients are amenable to surgery. These facts have prompted us to evaluate approaches including perioperative therapy modalities. METHODS: By means of a computer-supported search (MEDLINE, ASCO Proceedings) as well as a manual literature search, randomized clinical trials and meta-analyses evaluating preoperative therapy plus surgery in patients with resectable esophageal cancer were identified. RESULTS: Twenty-three randomized clinical studies and five meta analyses were identified. Interpretation--especially of the older studies- however, is hampered by the lack of a stringent application of modern examination techniques allowing for exact specification of tumor stage (resectable/locally advanced) as well as the inclusion of patients with different histological entities (squamous cell/adenocarcinoma), different surgical techniques and the low number of patients. There were no significant differences for the following perioperative therapy modalities regarding tumor-related mortality: preoperative radiotherapy versus postoperative radiotherapy, preoperative and postoperative radiotherapy versus postoperative radiotherapy (higher mortality in preoperative and postoperative radiotherapy arm), preoperative radiotherapy versus surgery, preoperative chemotherapy versus surgery, preoperative and postoperative chemotherapy versus surgery. In terms of 3-year mortality, statistically significant differences could be found for the following therapy modalities: preoperative radiochemotherapy versus surgery (preoperative therapy superior), preoperative chemotherapy versus preoperative radiotherapy (preoperative radiotherapy superior). CONCLUSION: Judging from these data, no clear recommendation for a standard multimodality approach outside clinical studies can be given for patients with resectable esophageal cancer. Neoadjuvant therapy, therefore, can only be recommended in centers with a maximum surgical expertise and in the context of multidisciplinary study protocols. PMID- 16220456 TI - [Liver metastases: incidence and histogenesis]. AB - Metastases are the most common malignant tumors of the liver. In the files of the Institute of Pathology of the University of Cologne 12,161 liver tissue cases are registered. Of them, 1,357 cases (11.2%) showed tumors or tumor like masses. Liver metastases of solid tumors were the largest group of the neoplasias with 611 cases (5.0%) followed by hepatocellular carcinoma (380 cases; 3.1%). Other entities were rare and include cholangiolar carcinoma (0.5%), vascular tumors (0.4%), lymphomas (0.4%), focal nodular hyperplasias (0.36%) and liver cell adenomas (0.23%). Adenocarcinomaa are the largest group of metastases with 400 cases (65.5%). 48.2% of this group were metastases of colorectal cancer, 13.5% of pancreatic cancer, 13% of breast cancer, 6.2% of gastric cancer, 4.5% of lung cancer and 3.7% of esophageal cancer. Neuroendocrine carcinomas are the second largest group with 16% of liver filiae. Other entities were rarely found. Metastases in cirrhotic livers are seldom. The gross findings, the histology, the differential diagnosis including immunohistochemistry and the value of the liver biopsy is discussed. PMID- 16220457 TI - [Does liver cirrhosis cause clinically relevant, but reversible chronic heart failure?]. PMID- 16220461 TI - [A novel KIT gene mutation results in piebaldism]. AB - OBJECTIVE: To detect gene mutation in proband and his mother from a family with piebaldism. METHODS: Diagnosis of a patient with piebaldism was validated by pathology, ultrastructural examination and the typical clinical manifestation. PCR and DNA sequencing were carried out to detect gene mutation of a family with piebaldism. RESULTS: G1833A transition in the KIT gene was found in the proband of the family with piebaldism. This mutation resulted in V604I substitution in KIT gene. No mutation was found in 100 normal individuals and other family members. CONCLUSION: The mutation of V604I is the cause of clinical phenotype of the family with piebaldism. PMID- 16220462 TI - Checking stationarity of the incidence rate using prevalent cohort survival data. AB - When survival data are collected as part of a prevalent cohort study with follow up, the recruited cases have already experienced their initiating event, say onset of a disease, and consequently the incidence process is only partially observed. Nevertheless, there are good reasons for interest in certain features of the underlying incidence process, for example whether or not it is stationary. Indeed, the well known relationship between incidence and prevalence, often used by epidemiologists, requires stationarity of the incidence rate for its validity. Also, the statistician can exploit stationarity of the incidence process by improving the efficiency of estimators in a prevalent cohort survival analysis. In addition, whether the incident rate is stationary is often in itself of central importance to medical and other researchers. We present here a necessary and sufficient condition for stationarity of the underlying incidence process, which uses only survival observations, possibly right censored, from a prevalent cohort study with follow-up. This leads to a simple graphical means of checking for the stationarity of the underlying incidence times by comparing the plots of two Kaplan-Meier estimates that are based on partially observed incidence times and follow-up survival data. We use our method to discuss the incidence rate of dementia in Canada between 1971 and 1991. PMID- 16220463 TI - The natural abundance of 13C, 15N, 34S and 14C in archived (1923-2000) plant and soil samples from the Askov long-term experiments on animal manure and mineral fertilizer. AB - The Askov field experiment (Denmark), established in 1894, provides a unique opportunity to examine long-term effects of animal manure and mineral fertilizer on soil organic matter quality and turnover. This sandy loam soil is classified as Alfisol (Typic Hapludalf). Soil C, N, S, 13C, 15N, 34S and 14C contents were measured in a selection of archived soil samples (1923, 1938, 1945, 1953, 1964, 1976, 1985, 1996 and 2000) from unfertilized (O), animal manure (1 AM) and mineral fertilizer (1 NPK) treatments. These treatments are imbedded in a four course crop rotation of winter cereals, root crops, spring cereals and a clover/grass mixture. The contents of C, N, S, 13C, 15N and 34S in selected crop samples (1953-1996) and in contemporary samples of animal feed and manure were also determined. Temporal soil nutrient and isotope trends between fertilizer treatments were significantly different, except for S content in 1 AM and 1 NPK. The total soil C and S was higher in 1 AM and 1 NPK than in the O treatment. The total soil N content (1 AM>1 NPK>O) and the delta15N content (1 AM>1 NPK and O) were also different. Analyses of plant, animal feed and manures confirmed that differences in soil 15N values were related to delta15N values of added source inputs. Soil and crop delta13C values were similar, but manures had slightly lower values. The variation of soil delta34S (and total S) from 1923 to 1996 was larger in the O than 1 AM and 1 NPK plots reflecting changes in atmospheric S inputs. The total contents of soil C, N and S were significantly correlated, but their isotopic signatures were not, suggesting that the C, N, S turnovers in soil are subject to different controls. The 14C content was generally higher in the 1 AM than 1 NPK and O, with bomb-14C incorporation modelling indicating that mean residence time (MRT) was ca. 170 years in the 1 AM, but closer to 250-290 years in the 1 NPK and O treatments. The measured trends in soil C and 14C during 1923 1996 were successfully modelled using the RothC model. The OM accumulation in the Askov soils was generally dominated by microbial decomposition products rather than by recalcitrant components of the various inputs. PMID- 16220464 TI - Bulk and compound-specific isotopic characterisation of illicit heroin and cling film. AB - Comparative analysis involves various but complementary methods and can be used for forensic intelligence purposes to group seizures of heroin into batches. Much forensic analysis now combines expertise in the traditional area of drugs investigation with a detailed understanding of supply, packaging, distribution, and drugs intelligence. It was the intention of this research to determine whether illicit heroin seizures and packaging material can be grouped according to isotopic compositions, and to explore factors that affect the isotopic compositions. In order to achieve these aims, 14 samples of seized heroin, thirteen provided by Avon and Somerset Constabulary (UK), were analysed by elemental analysis/isotope ratio mass spectrometry (EA/IRMS) and gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) for carbon and hydrogen isotopes. These tests elucidated that a combination of the delta13C, delta15N, delta18O and delta2H results from EA/IRMS is able to distinguish between most samples of bulk heroin. We speculate that the delta13C values of the alkaloids, obtained by GC/C/IRMS, give indications of different geographical or temporal origins of some of the heroin samples. GC/C/IRMS of the cutting agent, caffeine, provides a means to link dilution events. Fifteen retail cling film samples and seven cling film samples from heroin seizures were analysed by EA/IRMS. A multivariate comparison of the carbon, hydrogen and oxygen isotope ratios was able to distinguish between most of the samples. This technique enabled the cling films from the heroin to be grouped according to seizure. Three solvents were tested on two samples of cling film of known composition. Methanol and chloroform were both found to extract material from PVC and from non-PVC cling films. Water-treated PVC was indistinguishable from the untreated PVC and thus water was found to be the most suitable solvent when washing cling film prior to IRMS analysis. PMID- 16220465 TI - Stable isotope analysis of safety matches using isotope ratio mass spectrometry- a forensic case study. AB - Isotope ratio mass spectrometry (IRMS) was used to assess what contribution the technique could make towards the comparative analysis of matchstick samples within the 'normal' framework of a forensic investigation. A method was developed to allow the comparison of samples submitted as a result of an investigation, with the added advantage of rapid sample turn-around expected within this field. To the best of our knowledge this is the first time that wooden safety matches have been analysed using IRMS. In this particular case, bulk stable isotope analysis carrried out on a 'like-for-like' basis could demonstrate conclusively that matches seized from a suspect were different from those collected at the scene of crime. The maximum delta13C variability observed within one box was 2.5 per thousand, which, in conjunction with the error of measurement, was regarded to yield too wide an error margin as to permit differentiation of matchsticks based on 13C isotopic composition alone given that the 'natural' 13C abundance in wood ranges from -20 to -30 per thousand. However, from the delta2H values obtained for crime scene matches and seized matches of -114.5 per thousand and 65 per thousand, respectively, it was concluded that the matches seized were distinctly different from those collected at the crime scene. PMID- 16220466 TI - Photodissociation of singly protonated peptides at 193 nm investigated with tandem time-of-flight mass spectrometry. AB - Photodissociation at 193 nm of some singly protonated peptides generated by matrix-assisted laser desorption/ionization was investigated using tandem time-of flight mass spectrometry. For peptides with arginine at the C-terminus, x, upsilon, and w fragment ions were generated preferentially while a and d fragment ions dominated for peptides with arginine at the N-terminus. These are the same characteristics as photodissociation at 157 nm reported previously. Overall, the photodissociation spectra obtained at 157 and 193 nm were strikingly similar. PMID- 16220467 TI - Electrospray ionization mass spectrometry study on dipeptide 4-chlorobutyl ester produced from refluxing of amino acid with phosphoryl chloride in tetrahydrofuran. AB - Phosphoryl chloride was able to promote the formation of peptide and the ringopening of tetrahydrofuran (THF) followed by a sequence of successive reactions by simply refluxing. ESI multistage tandem mass spectrometry was applied to trace the reaction and elucidate the product structures, dipeptide 4 chlorobutyl ester. PMID- 16220468 TI - Nonacid cleavable detergents applied to MALDI mass spectrometry profiling of whole cells. AB - Although cleavable detergents were first synthesized a number of years ago, they have only recently been successfully applied to problems involving biological molecules. Recent reports have demonstrated that these compounds are useful for applications involving both 2D PAGE and mass spectrometry. However, most cleavable surfactants have utilized acid-labile functional groups to affect cleavage. In applications where extreme pH is required, acid cleavable detergents have limited usefulness. We report the synthesis of fluoride cleavable silane compounds and photolabile cinnamate esters as cleavable detergents having alternative cleavage chemistries than previously reported cleavable detergents. These compounds were applied to whole cell analysis using MALDI mass spectrometry, and it was demonstrated that their use results in an increase in the number of proteins analyzed by increasing protein solubility. PMID- 16220469 TI - Assessment of pepper spray product potency in Asian and Caucasian forearm skin using transepidermal water loss, skin temperature and reflectance colorimetry. AB - Historically, pepper spray product potency has been established using a taste test evaluation. A taste test is subjective and may not be appropriate for assessing pepper potency in skin. The current study evaluated chemically diverse pepper sprays in human forearm skin using three objective, noninvasive parameters: transepidermal water loss, skin surface temperature and erythema, as a means for assessing dermal pharmacology, toxicology and product potency. Five commercial pepper spray products containing various capsaicinoid analogs at various concentrations were evaluated in duplicate on volar forearms of six Caucasians and six Asians using a 10 min exposure. Mean surface skin temperature, transepidermal water loss results were highly variable and therefore did not demonstrate dose responsive behavior to increasing capsaicinoid concentrations. Erythema, as measured by increases in a* (reflected light in the red-to-green color spectrum) of the L*a*b* uniform color scale, was superior among parameters evaluated in discriminating pepper spray potency and correlated well with the relative and total capsaicinoid concentration in the products. Products containing greater than 16 mg ml(-1) capsaicinoid concentration produced greater erythema responses in Caucasians than Asians. Asians responded greater to the synthetic analog, nonivamide, than to mixtures of capsaicinoids, while Caucasians responded equally to both capsaicinoid analogs. Thus, pepper spray product potency in human skin reflects the total capsaicinoid concentration, the specific capsaicin analog(s) present, and the race of the individual exposed. The finding that the reflectance colorimeter a* scale can differentiate these parameters in skin will have a significant impact on evaluating the use and efficacy of pepper spray products in humans. PMID- 16220470 TI - Comparison of predictive values of two diagnostic tests from the same sample of subjects using weighted least squares. AB - Screening and diagnostic tests are important in disease prevention or control. The predictive values of positive and negative (PPV and NPV) test results are two of four operational characteristics of a screening test. We review an existing method based on the generalized estimating equation (GEE) methodology for comparing predictive values from the same sample of subjects and propose two Wald test statistics derived from the weighted least squares (WLS) method for the analysis of categorical data. Using these results, we propose sample size calculation formulae for this problem. Simulation studies are conducted to compare the performances of the two Wald test statistics (one based on the difference of two PPVs or NPVs, another based on the logarithm of the ratio of two PPVs or NPVs) and the score/Wald test statistic derived from GEE. We recommend using the difference-based WLS approach. PMID- 16220472 TI - Interval estimation of the mean response in a log-regression model. AB - A standard approach to the analysis of skewed response data with concomitant information is to use a log-transformation to normalize the distribution of the response variable and then conduct a log- regression analysis. However, the mean response at original scale is often of interest. El-Shaarawi and Viveros developed an interval estimation of the mean response of a log-regression model based on large sample theory. There is however very little information available in the literature on constructing such estimates when the sample size is small. In this paper, we develop a small-sample corrected interval by using the likelihood-based inference method developed by Barndorff-Nielson and Fraser et al. Simulation results show that the proposed interval provides almost exact coverage probability, even for small samples. PMID- 16220471 TI - Analysis of the renal transplant waiting list in the Pais Valencia (Spain). AB - In this paper we analyse the renal transplant waiting list of the Pais Valencia in Spain, using Queueing theory. The customers of this queue are patients with end-stage renal failure waiting for a kidney transplant. We set up a simplified model to represent the flow of the customers through the system, and perform Bayesian inference to estimate parameters in the model. Finally, we consider several scenarios by tuning the estimations achieved and computationally simulate the behaviour of the queue under each one. The results indicate that the system could reach equilibrium at some point in the future and the model forecasts a slow decrease in the size of the waiting list in the short and middle term. PMID- 16220473 TI - Curious phenomena in Bayesian adjustment for exposure misclassification. AB - Many epidemiologic investigations involve some discussion of exposure misclassification, but rarely is there an attempt to adjust for misclassification formally in the statistical analysis. Rather, investigators tend to rely on intuition to comment qualitatively on how misclassification might impact their findings. We point out several ways in which intuition might fail, in the context of unmatched case-control analysis with non-differential exposure misclassification. Particularly, we focus on how intuition can conflict with the results of a Bayesian analysis that accounts for the various uncertainties at hand. First, the Bayesian adjustment for misclassification can weaken the evidence about the direction of an exposure-disease association. Second, admitting uncertainty about the misclassification parameters can lead to narrower interval estimates concerning the association. We focus on the simple setting of unmatched case-control analysis with binary exposure and without adjustment for confounders, though much of our discussion should be relevant more generally. PMID- 16220474 TI - A multivariate random-effects model with restricted parameters: application to assessing radiation therapy for brain tumours. AB - In clinical studies, multiple endpoints are often measured for each patient longitudinally. The multivariate random-effects or random coefficient model has been a useful method for analysis. However, medical research problems may impose restrictions on the model parameters of interests. For example, in a paediatric brain tumour study on radiation therapy, there is a natural ordering in the white matter relaxation time of brain tissues among different regions surrounding the primary tumour, i.e. the closer a specific region of brain tissues is to the centre of primary tumour, the shorter is the relaxation time. Such parameter constraints should be accounted for in the analysis. This article proposes a class of multivariate random coefficient models with restricted parameters and derives its maximum likelihood estimates (MLE). We propose a modified EM algorithm for the quadratic optimalization with linear inequality constraints necessary in deriving the MLE. The method is applied to analysing the paediatric brain tumour study. PMID- 16220475 TI - Power to detect clinically relevant carry-over in a series of cross-over studies. AB - The potential for carry-over effects is an important consideration in the design of any cross-over study, and can cause an investigator to abandon the design altogether. In cross-over studies, carry-over is the lingering effect of a treatment into the subsequent period. Carry-over effects are differences in the extent of the carry-over between the treatments under consideration. It is well known that the test for carry-over effects in individual studies has low power. Empirical evidence of carry-over effects, or the absence of carry-over effects, could be useful for investigators considering the design. Here we develop methods for expressing the power to detect carry-over as a function of the power to detect a clinically relevant treatment effect. Our results suggest that for two treatment, two-period cross-over studies the power to detect clinically relevant carry-over effects is often less than 15 per cent, and the number of studies needed to differentiate this effect from the type I error rate of 10 per cent is prohibitive. For the three-treatment three-period cross-over design, the power to detect carry-over effects was larger than for the two-period study, but still approached the type I error rate in a number of cases. Unequivocal conclusions about the absence of carry-over effects based on collections of hypothesis tests appear unlikely. Similar findings are presented for bioequivalence studies. For bioequivalence studies, small carry-over effects (e.g. 12.5 per cent of the treatment effect) can seriously inflate the type I error rate, particularly when the power to detect equivalence is high. PMID- 16220476 TI - Bivariate isotonic design for dose-finding with ordered groups. AB - We consider the problem of dose-finding where subjects can be stratified into two populations with possibly different susceptibility to toxicity. The goal is to find the maximally tolerated dose for each population. We propose a non parametric design for this problem. Toxicity is estimated using the bivariate isotonic regression estimator. The new design is compared with the two-sample continual reassessment method. PMID- 16220477 TI - Assessment on homogeneity tests for kappa statistics under equal prevalence across studies in reliability. AB - In this paper, we assess the performance of homogeneity tests for two or more kappa statistics when prevalence rates across reliability studies are assumed to be equal. The likelihood score method and the chi-square goodness-of-fit (GOF) test provide type 1 error rates that are satisfactorily close to the nominal level, but a Fleiss-like test is not satisfactory for small or moderate sample sizes. Simulations show that the score test is more powerful than the chi-square GOF test and the approximate sample size required for a specific power of the former is substantially smaller than the latter. In addition, the score test is robust to deviations from the equal prevalence assumption, while the GOF test is highly sensitive and it may give a grossly misleading type 1 error rate when the assumption of equal prevalence is violated. We conclude that the homogeneity score test is the preferred method. PMID- 16220478 TI - An adaptive dose-finding design incorporating both toxicity and efficacy. AB - Novel therapies are challenging the standards of drug development. Agents with specific biologic targets and limited toxicity require novel designs to determine doses to be taken forward into larger studies. In this paper, we describe an approach that incorporates both toxicity and efficacy data into the estimation of the biologically optimal dose of an agent in a phase I trial. The approach is based on the flexible continuation-ratio model, and uses straightforward optimal dose selection criteria. Dose selection is based on all patients treated up until that time point, using a continual reassessment method approach. Dose-outcome curves considered include monotonically increasing, monotonically decreasing, and unimodal curves. Our simulation studies demonstrate that the proposed design, which we call TriCRM, has favourable operating characteristics. PMID- 16220479 TI - The sequential analysis of repeated binary responses: a score test for the case of three time points. AB - In this paper a robust method is developed for the analysis of data consisting of repeated binary observations taken at up to three fixed time points on each subject. The primary objective is to compare outcomes at the last time point, using earlier observations to predict this for subjects with incomplete records. A score test is derived. The method is developed for application to sequential clinical trials, as at interim analyses there will be many incomplete records occurring in non-informative patterns. Motivation for the methodology comes from experience with clinical trials in stroke and head injury, and data from one such trial is used to illustrate the approach. Extensions to more than three time points and to allow for stratification are discussed. PMID- 16220480 TI - Causal conclusions are most sensitive to unobserved binary covariates. AB - There is a rich literature that considers whether an observed relation between treatment and response is due to an unobserved covariate. In order to quantify this unmeasured bias, an assumption is made about the distribution of this unobserved covariate; typically that it is either binary or at least confined to the unit interval. In this paper, this assumption is relaxed in the context of matched pairs with binary treatment and response. One might think that a long tailed unobserved covariate could do more damage. Remarkably that is not the case: the most harm is done by a binary covariate, so the case commonly considered in the literature is most conservative. This has two practical consequences: (i) it is always safe to assume that an unobserved covariate is binary, if one is content to make a conservative statement; (ii) when another assumption seems more appropriate, say normal covariate, there will be less sensitivity than with a binary covariate. This assumption implies that it is possible that a relation between treatment and response that is sensitive to unmeasured bias (if the unobserved covariate is dichotomous), ceases to be sensitive if the unobserved covariate is normally distributed. These ideas are illustrated by three examples. It is important to note that the claim in this paper applies to our specific setting of matched pairs with binary treatment and response. Whether the same conclusion holds in other settings is an open question. PMID- 16220481 TI - Using the ROC curve for gauging treatment effect in clinical trials. AB - Non-parametric procedures such as the Wilcoxon rank-sum test, or equivalently the Mann-Whitney test, are often used to analyse data from clinical trials. These procedures enable testing for treatment effect, but traditionally do not account for covariates. We adapt recently developed methods for receiver operating characteristic (ROC) curve regression analysis to extend the Mann-Whitney test to accommodate covariate adjustment and evaluation of effect modification. Our approach naturally extends use of the Mann-Whitney statistic in a fashion that is analogous to how linear models extend the t-test. We illustrate the methodology with data from clinical trials of a therapy for Cystic Fibrosis. PMID- 16220482 TI - Effect measures in non-parametric regression with interactions between continuous exposures. AB - In many biomedical studies, interest is often attached to calculating effect measures in the presence of interactions between two continuous exposures. Traditional approaches based on parametric regression are limited by the degree of arbitrariness involved in transforming these exposures into categorical variables or imposing a parametric form on the regression function. In this paper, we present: (a) a flexible non-parametric method for estimating effect measures through generalized additive models including interactions; and (b) bootstrap techniques for (i) testing the significance of interaction terms, and (ii) constructing confidence intervals for effect measures. The validity of our methodology is supported by simulations, and illustrated using data from a study of possible risk factors for post-operative infection. This application revealed a hitherto unreported effect: for patients with high plasma glucose levels, increased risk is associated, not only with low, but also with high percentages of lymphocytes. PMID- 16220483 TI - Modelling tumour biology-progression relationships in screening trials. AB - There has been some recent work in the statistical literature for modelling the relationship between tumour biology properties and tumour progression in screening trials. While non-parametric methods have been proposed for estimation of the tumour size distribution at which metastatic transition occurs, their asymptotic properties have not been studied. In addition, no testing or regression methods are available so that potential confounders and prognostic factors can be adjusted for. We develop a unified approach to non-parametric and semi-parametric analysis of modelling tumour size-metastasis data in this article. An association between the models considered by previous authors with survival data structures is discussed. Based on this relationship, we develop non parametric testing procedures and semi-parametric regression methodology of modelling the effect of size of tumour on the probability at which metastatic transitions occur in two situations. Asymptotic properties of these estimators are provided. The proposed methodology is applied to data from a screening study in lung cancer. PMID- 16220484 TI - Comparison of case-deletion diagnostic methods for Cox regression. AB - Case-deletion diagnostics are a routine component of regression analysis since they identify unusual observations that substantially affect parameter estimates. The exact approach is to compute the change in each regression parameter by dropping that individual and refitting the model. Repeating a Cox regression for the removal of each individual is very time consuming and therefore not done in practice. The two methods commonly used to approximate the exact case-deletion change for Cox regression are the empirical influence function approach and the covariate-vector augmentation approach. This paper reports the results of a simulation study on how well these methods estimate the exact change in a parameter estimate when deleting a known outlier or a known non-outlier. Additionally, we investigate how well these methods correctly identify outliers and non-outliers. The covariate augmentation approach clearly outperformed the influence function approach in these simulations. PMID- 16220485 TI - An efficient design for a study comparing two drugs, their combination and placebo. AB - A novel design for a study comparing two drugs (A and B), their combination (AB) and placebo (P) in a stable disease such as chronic asthma is proposed. The primary objectives of the trial are to compare A (a new drug) with placebo and AB with B (an old drug). Other between-treatment comparisons are secondary. The new design consists of the first 2 sequences of a single 4 x 4 Latin square: (A, AB, P, B|P, B, A, AB|AB, A, B, P|B, P, AB, A). The washout period between A and AB and that between P and B are eliminated based on the assumptions that there is no carry-over effect from placebo and that the effect of the combination would be approximately same at steady state either starting the 2 drugs at the same time or adding one later. Equal period effect for Periods 1 and 2, and for Periods 3 and 4 are also assumed for all treatment effects being estimable. Advantages and disadvantages of this design were evaluated and compared with other possible designs. Compared with other possible designs, the new design is more efficient. For example, the new design requires less than (1/4) of the sample size a parallel design would need and it requires (2/3) of the sample size a 4 treatment, 3-period incomplete block design would need for the same power at the same detectable difference. It has a similar efficiency to a 4-treatment, 4 period cross-over design but with a shorter duration. In this design, all between treatment comparisons are based on direct within-subject information. The new design could also potentially decrease the probability of patient's drop-outs and use of rescue medication due to lack of efficacy during the placebo periods by reducing the number of placebo washout periods. The benefits reaped from the new design seem to outweigh its risks, which include potential bias for the secondary between-treatment comparisons if the additional assumption about period effects are not satisfied and the incapability of directly assessing the onset of action of the combination of 2 drugs. PMID- 16220486 TI - Estimation of time-dependent area under the ROC curve for long-term risk prediction. AB - Sensitivity, specificity, and area under the ROC curve (AUC) are often used to measure the ability of survival models to predict future risk. Estimation of these parameters is complicated by the fact that these parameters are time dependent and by the fact that censoring affects their estimation just as it affects estimation of survival curves or coefficients of survival regression models. The authors present several estimators that overcome these complications. One approach is a recursive calculation over the ordered times of events, analogous to the Kaplan-Meier approach to survival function estimation. Another is to first apply Bayes' theorem to write the parameters of interest in terms of conditional survival functions that are then estimated by survival analysis methods. Simulation studies demonstrate that the proposed estimators perform well in practical situations, when compared with an estimator (c-statistic, from logistic regression) that ignores time. An illustration with data from a cardiovascular follow-up study is provided. PMID- 16220487 TI - Random effects logistic models for analysing efficacy of a longitudinal randomized treatment with non-adherence. AB - We present a random effects logistic approach for estimating the efficacy of treatment for compliers in a randomized trial with treatment non-adherence and longitudinal binary outcomes. We use our approach to analyse a primary care depression intervention trial. The use of a random effects model to estimate efficacy supplements intent-to-treat longitudinal analyses based on random effects logistic models that are commonly used in primary care depression research. Our estimation approach is an extension of Nagelkerke et al.'s instrumental variables approximation for cross-sectional binary outcomes. Our approach is easily implementable with standard random effects logistic regression software. We show through a simulation study that our approach provides reasonably accurate inferences for the setting of the depression trial under model assumptions. We also evaluate the sensitivity of our approach to model assumptions for the depression trial. PMID- 16220488 TI - Interval estimation for treatment effects using propensity score matching. AB - In causal studies without random assignment of treatment, causal effects can be estimated using matched treated and control samples, where matches are obtained using estimated propensity scores. Propensity score matching can reduce bias in treatment effect estimators in cases where the matched samples have overlapping covariate distributions. Despite its application in many applied problems, there is no universally employed approach to interval estimation when using propensity score matching. In this article, we present and evaluate approaches to interval estimation when using propensity score matching. PMID- 16220489 TI - Estimation in flexible two stage designs. AB - Adaptive test designs for clinical trials allow for a wide range of data driven design adaptations using all information gathered until an interim analysis. The basic principle is to use a test statistics which is invariant with respect to the design adaptations under the null hypothesis. This allows for a control of the type I error rate for the primary hypothesis even for adaptations not specified a priori in the study protocol. Estimation is usually another important part of a clinical trial, however, is more difficult in adaptive designs. In this research paper we give an overview of point and interval estimates for flexible designs and compare methods for typical sample size rules. We also make some proposals for confidence intervals which have nominal coverage probability also after an unforeseen design adaptation and which contain the maximum likelihood estimate and the usual unadjusted confidence interval. PMID- 16220490 TI - A comparison of propensity score methods: a case-study estimating the effectiveness of post-AMI statin use. AB - There is an increasing interest in the use of propensity score methods to estimate causal effects in observational studies. However, recent systematic reviews have demonstrated that propensity score methods are inconsistently used and frequently poorly applied in the medical literature. In this study, we compared the following propensity score methods for estimating the reduction in all-cause mortality due to statin therapy for patients hospitalized with acute myocardial infarction: propensity-score matching, stratification using the propensity score, covariate adjustment using the propensity score, and weighting using the propensity score. We used propensity score methods to estimate both adjusted treated effects and the absolute and relative risk reduction in all cause mortality. We also examined the use of statistical hypothesis testing, standardized differences, box plots, non-parametric density estimates, and quantile-quantile plots to assess residual confounding that remained after stratification or matching on the propensity score. Estimates of the absolute reduction in 3-year mortality ranged from 2.1 to 4.5 per cent, while estimates of the relative risk reduction ranged from 13.3 to 17.0 per cent. Adjusted estimates of the reduction in the odds of 3-year death varied from 15 to 24 per cent across the different propensity score methods. PMID- 16220491 TI - The effects of non-compliance on intent-to-treat analysis of equivalence trials. AB - The standard approach for analysing a randomized clinical trial is based on intent-to-treat (ITT) where subjects are analysed according to their assigned treatment group regardless of actual adherence to the treatment protocol. For therapeutic equivalence trials, it is a common concern that an ITT analysis increases the chance of erroneously concluding equivalence. In this paper, we formally investigate the impact of non-compliance on an ITT analysis of equivalence trials with a binary outcome. We assume 'all-or-none' compliance and independence between compliance and the outcome. Our results indicate that non compliance does not always make it easier to demonstrate equivalence. The direction and magnitude of changes in the type I error rate and power of the study depend on the patterns of non-compliance, event probabilities, the margin of equivalence and other factors. PMID- 16220492 TI - Inference for a family of survival models encompassing the proportional hazards and proportional odds models. AB - For survival data regression, the Cox proportional hazards model is the most popular model, but in certain situations the Cox model is inappropriate. Various authors have proposed the proportional odds model as an alternative. Yang and Prentice recently presented a number of easily implemented estimators for the proportional odds model. Here we show how to extend the methods of Yang and Prentice to a family of survival models that includes the proportional hazards model and proportional odds model as special cases. The model is defined in terms of a Box-Cox transformation of the survival function, indexed by a transformation parameter rho. This model has been discussed by other authors, and is related to the Harrington-Fleming G(rho) family of tests and to frailty models. We discuss inference for the case where rho is known and the case where rho must be estimated. We present a simulation study of a pseudo-likelihood estimator and a martingale residual estimator. We find that the methods perform reasonably. We apply our model to a real data set. PMID- 16220493 TI - Partial questionnaire designs, questionnaire non-response, and attributable fraction: applications to adult onset asthma. AB - The attributable fraction (AF) is often used to explore the policy implications of an association between a disease and an exposure. To date, there have been no proposed estimators of AF in the context of partial questionnaire designs (PQD). The PQD, first proposed in a public health context by Wacholder is often used to enhance response rates in questionnaires. It involves eliciting responses from each subject on preassigned subsets of questions, thereby reducing the burden of response. We propose a computationally efficient method of estimating logistic (or more generally, binary) regression parameters from a PQD model where there is non-response to the questionnaire and the rates of non-response differ between sub-populations. Assuming a log-linear model for the distribution of missing covariates, we employ the methods of Wacholder to motivate consistent estimating equations, and weight each subject's contribution to the estimating function by the inverse probability of responding to the questionnaire. We also propose techniques for goodness-of-fit to assist in model selection. We then use the PQD regression estimates to derive an estimate of AF similar to that proposed by Bruzzi. Finally, we demonstrate our methods using data obtained from a study on adult occupational asthma, conducted within a Massachusetts HMO. Although we concentrate on a particular type of missing data mechanism, other missing data techniques can be incorporated into AF estimation in a similar manner. PMID- 16220495 TI - Analysis of incomplete longitudinal binary data using multiple imputation. AB - We propose a propensity score-based multiple imputation (MI) method to tackle incomplete missing data resulting from drop-outs and/or intermittent skipped visits in longitudinal clinical trials with binary responses. The estimation and inferential properties of the proposed method are contrasted via simulation with those of the commonly used complete-case (CC) and generalized estimating equations (GEE) methods. Three key results are noted. First, if data are missing completely at random, MI can be notably more efficient than the CC and GEE methods. Second, with small samples, GEE often fails due to 'convergence problems', but MI is free of that problem. Finally, if the data are missing at random, while the CC and GEE methods yield results with moderate to large bias, MI generally yields results with negligible bias. A numerical example with real data is provided for illustration. PMID- 16220494 TI - Secondary analysis of case-control data. AB - We extend the discussion of Lee et al. and others on methods for performing secondary analyses of case-control sampled data and carry out an extensive investigation of efficiency and robustness. We find that, with the exception of the 'analyse-the-controls-only' strategy for populations in which cases are rare, ad hoc methods in common usage often lead to extremely misleading conclusions and that it is not possible to tell in advance when this will happen. Weighted likelihood and semi-parametric maximum likelihood methods are justified theoretically. We find that semi-parametric maximum likelihood can be as much as twice as efficient as the weighted method, but is subject to bias in estimating parameters of interest when the nuisance models this method requires have been mis-specified. The weighted method needs no nuisance models and thus is robust in this regard, but we cannot tell when it is going to be very inefficient without sophisticated modelling as through the SPML method. Practitioners should routinely use both methods and will often have to weigh up the practical consequences of severe inefficiency and lack of robustness in the context of their enquiries. PMID- 16220496 TI - A new classification rule for incomplete doubly multivariate data using mixed effects model with performance comparisons on the imputed data. AB - A mixed effects model, enhanced by a Kronecker product structure for the residual variance-covariance matrix, is used in conjunction with a discriminant analysis technique, to devise a new statistical classification method on incomplete doubly multivariate data. The proposed method is efficient in small scale clinical trials that use relatively few patients. The new classification method is also applied to multiply imputed data sets. The misclassification error rates (MERs) are compared in order to investigate the effectiveness of the new classification rule on an incomplete data set. The classification method is applied to a real data set. The error rates on the incomplete data set are found to be much less than the median error rate on the multiply imputed data sets. Non-parametric methods, such as kernel method and k-nearest neighbourhood method, are also applied to multiply imputed data sets. Results illustrating the advantages of the new classification method over classic non-parametric classification methods are presented. PMID- 16220497 TI - A unifying approach for surrogate marker validation based on Prentice's criteria. AB - Part of the recent literature on the evaluation of surrogate endpoints starts from a multi-trial approach which leads to a definition of validity in terms of the quality of both trial-level and individual-level association between a potential surrogate and a true endpoint, Buyse et al. These authors proposed their methodology based on the simplest cross-sectional case in which both the surrogate and the true endpoint are continuous and normally distributed. Different variations to this theme have been implemented for binary responses, times to event, combinations of binary and continuous endpoints, etc. However, a drawback of this methodology is that different settings have led to different definitions to quantify the association at the individual-level. In the longitudinal setting; Alonso et al. defined a class of canonical correlation functions that can be used to study surrogacy at the trial and individual-level. In the present work, we propose a new approach to evaluate surrogacy in the repeated measurements framework, we also show the connection between this proposal and the previous ones reported in the literature. Finally, we extend this concept to the non-normal case using the so-called 'likelihood reduction factor' (LRF) a new validation measure based on some of the Prentice's criteria. We apply the previous methodology using data from two clinical studies in psychiatry and ophthalmology. PMID- 16220498 TI - Estimating HIV hazard rates from cross-sectional HIV prevalence data. AB - We propose a method of estimating the HIV hazard rate in a population when only cross-sectional data measuring HIV-prevalence are available. In a cross-sectional study, individuals who test positive are left-censored since we know only that the transition time is smaller than the monitoring time. Individuals who test negative are right-censored since we know only that the transition time is greater than the monitoring time. The transition time is not observed directly. For our method, we assume the hazard rate is well-defined after the time we start screening. We assume further the mortality rates for HIV-positive and HIV negative individuals are known. The method is illustrated using HIV-prevalence data collected over four years among women in Cape Town, South Africa. PMID- 16220499 TI - Carbon isotope analysis of bulk keratin and single amino acids from British and North American hair. AB - The reconstruction of ancient diets using isotopic measurements of bone collagen, and other tissues, which survive in archaeological contexts, relies on known isotopic relationships between diet and body tissues. Examination of these relationships often requires the study of modern human and animal subjects. While hair keratin can act as a useful proxy for bone collagen in isotopic studies on living humans, where it is inappropriate to sample tissues such as collagen, it can, in addition, act as a chronological indicator of dietary change. This study investigates hair keratin delta13C values from current residents of the UK and the USA. Residents in the USA showed a clear bulk hair delta13C enrichment of approximately 3 per thousand over UK individuals, attributed to an elevated C4 dietary input from maize fed to livestock in North America. The keratin delta13C of subjects who moved between the UK and USA showed a pronounced change after relocation, taking approximately 4 months to reach isotopic equilibrium. To investigate these differences further, we measured delta13C values of dispensable and indispensable amino acids as a group, and selected individual amino acids. As a group, enrichment of dispensable amino acids compared with indispensable amino acids occurred in samples from both continents, averaging 7.2 per thousand in the UK and 7.9 per thousand in the USA. Dispensable and indispensable amino acids, as well as all individual amino acids measured, were enriched in samples from the USA compared with those from the UK. PMID- 16220500 TI - Advances in coupling a commercial total organic carbon analyser with an isotope ratio mass spectrometer to determine the isotopic signal of the total dissolved nitrogen pool. AB - A new method has been developed to analyse 15N of the total dissolved nitrogen (TDN) pool. The method operates on a commercial total organic carbon (TOC) analyser coupled to an elemental analyser/isotope ratio mass spectrometer (EA IRMS). Nitrogen compounds are combusted to nitric oxide (NO) and nitrogen dioxide (NO2) by high-temperature catalytic oxidation (HTCO), after which the NOx gas is transferred to an EA-IRMS for isotopic nitrogen analysis. The system is described, including five modifications of the system in order to overcome analytical problems. First, flow paths were modified to run both systems on helium as carrier gas, while complete sample oxidation was maintained. Secondly, the catalyst structure was adapted to allow high injection volumes at the given backpressures delivered by the EA system. Thirdly, we installed a Permapure dehumidification system as the standard Peltier element did not satisfy dehumidification requirements. Finally, we prevented the inflow of atmospheric nitrogen into the system. In a final stage, we are planning to automate the coupled system in order to run a continuous batch of up to 60 samples. We have obtained satisfactory results on the accuracy and precision of 180+/-1 per thousand potassium nitrate samples (IAEA, USGS-32). Running a batch of five samples resulted in a mean isotopic value of 178.8 per thousand with a standard deviation of 2.8 per thousand. Some important issues could not yet be addressed here, and will have to be evaluated once the system is running on a continuous base. However, the results appear promising and this system has the potential to become a method for TD15N analysis. An appropriate TD15N analysis method might open new challenges in aquatic and terrestrial ecosystem nitrogen studies, including a more comprehensive study of the dissolved organic nitrogen pool. PMID- 16220501 TI - Influence of initial charge state on fragmentation patterns for noncovalent drug/DNA duplex complexes. AB - The charge state-dependent dissociation of various DNA duplexes and drug/duplex complexes has been investigated using collisionally activated dissociation (CAD) in a quadrupole ion trap mass spectrometer (QIT-MS). Several non-self complementary 14-residue oligonucleotides were employed, in addition to an array of known DNA-interactive ligands, including the intercalators daunomycin and nogalamycin, as well as the minor groove binding agents distamycin, netropsin, 4',6-diamidino-2-phenylindole, and Hoechst 33342. In general, the dissociation pathways exhibited by both the duplexes and the drug/duplex complexes were found to be markedly sensitive to initial charge state. Time- and activation voltage independent duplex strand separation predominated for higher charge states, which was interpreted to be a result of internal Coulombic repulsion or partial unzipping in the interface, while time- and activation voltage-dependent covalent cleavage predominated for lower charge states. The identity of the drug and the sequence of the duplex were both found to affect the competition between different dissociation processes. The dissociation pathways for the lower charge state complexes are probably more reflective of specific drug-DNA interactions because Coulombic and/or conformational effects are less marked for these precursors. PMID- 16220502 TI - The effect of growth rate on tissue-diet isotopic spacing in rapidly growing animals. An experimental study with Atlantic salmon (Salmo salar). AB - The difference in isotopic composition between a consumer's tissues and that of its diet is a critical aspect of the use of stable isotope analyses in ecological and palaeoecological studies. In a controlled feeding experiment with the Atlantic salmon, Salmo salar, we demonstrate for the first time that the value of tissue-diet isotope spacing in nitrogen in a growing animal is not constant, but varies inversely with growth rate. The value of tissue-diet isotopic spacing in N reflects N use efficiency. Thus, in salmon, growth rate is accompanied by, or requires, increased N use efficiency. The total range in tissue-diet isotopic spacing in N seen in the experimental population of 25 fish was 1 per thousand, approximately 50% of the total trophic shift. Mean equilibrium tissue-diet isotopic spacing (+/-standard deviation) in salmon averaged 2.3 per thousand (+/ 0.3 per thousand) and 0.0 per thousand (+/-0.3 per thousand) for N in muscle and liver, respectively, and 2.1 per thousand (+/-0.1 per thousand) and 1.6 per thousand (+/-0.3 per thousand) for C in muscle and liver, respectively. Feeding with a mixed dietary source (wheat and fish-meal origin) resulted in tissue-diet isotopic fractionation in both C and N due to the differential digestibility of food components with distinct isotopic composition. The rate of change in isotopic composition of S. salar tissues was dominated by growth, but the estimated contribution of metabolic turnover to change in tissue N was relatively high for an ectothermic animal at ca. 20-40%. The estimated half-life for metabolic turnover of the tissue N pool was ca. 4 months in both muscle and liver tissue. This is the first study to demonstrate a direct relationship between tissue-diet isotopic spacing in N and growth rate and adds to the growing list of factors known to influence the level of isotopic separation between a consumer's tissue and that of its diet. PMID- 16220503 TI - Multiple losses of neutral C14H14 in the tandem mass spectrometry of several perbenzyl ether intermediates in the synthesis of green tea constituents. AB - Electrospray and matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry (MS/MS) experiments were used to investigate an unusual fragmentation in collision-induced dissociation (CID) of sodiated and potassiated perbenzyl ether intermediates obtained in the total synthesis of gallate ester constituents of green tea. Prominent fragments correspond to multiple sequential losses of neutral C14H14 that were not observed in the protonated and ammoniated species, that instead present fragment ion series in which members are separated by C7H6. High-resolution MALDI quadrupole time-of-flight (Q-TOF) and electrospray Fourier transform mass spectrometry (FTMS) were used to confirm elemental compositions of these and related ions. PMID- 16220504 TI - Differentiation of Boc- alpha,beta- and beta,alpha-peptides and a pair of diastereomeric beta,alpha-dipeptides by positive and negative ion electrospray tandem mass spectrometry (ESI-MS/MS). AB - Positive and negative ion electrospray ionization (ESI) tandem mass spectral study of a new series of hybrid peptides, viz, BocN-alpha,beta-peptides and BocN beta,alpha-peptides, synthesized from C-linked carbo-beta3-amino acids [Caa (S)] and L-Ala has been carried out. The alpha,beta-peptides have been differentiated from beta,alpha-peptides by the collision-induced dissociation (CID) of [M + H]+ and [M - H]- ions in positive and negative ion ESI-MS respectively. The fragment ion [M + H - C(CH3)3 + H]+ formed from [M + H]+ ions by the loss of 2-methyl-prop 2-ene in alpha,beta-peptides with L-Ala at the N-terminus is insignificant or totally absent for beta,alpha-peptides which have the Caa (S) at N-terminus. The fragment ion [M - H-C(CH3)3OH - HNCO]- formed from [M - H]- of beta,alpha-peptide acids is totally absent for alpha,beta-peptide acids. This has been attributed to the absence of the beta-methylene group in alpha,beta-peptides, and the participation of the beta-methylene group in the loss of HNCO in beta,alpha peptide acids is confirmed by the deuteration experiments. The CID of [M + H-Boc + H]+ ions of these peptides also produce characteristic fragmentation. In the CID spectra of alpha,beta-peptides, the b(n)+ ions and the resulting y(n)+ ions occur at a mass difference of 243 and 71 Da corresponding to the successive losses of Caa and L-Ala, whereas a mass difference of 71 and 243 Da is observed for beta,alpha-peptides. In contrast to the CID of protonated peptides, the CID of [M - H]- ions of the alpha,beta- and beta,alpha-peptide acids do not give b(n) ions and show abundant z(n) (-) ions. Further, a pair of diastereomeric dipeptide esters and acids have been distinguished by the CID of [M + H]+ ions. The loss of 2-methyl-prop-2-ene is more pronounced for Boc-NH-Caa(R)-D-Ala-OCH3 (21) and Boc-NH-Caa(R)-D-Ala-OH (23) with Caa (R) at the N-terminus, whereas it is totally absent for Boc-NH-Caa (S)-D-Ala-OCH3 (22) and Boc-NH-Caa(S)-D-Ala-OH (24) peptides, which have Caa (S) at the N-terminus. Thus, on the basis of our previous and present studies, we propose that the CID of [M + H]+ ions provides a simple and useful method for distinguishing the configuration of Caa (S or R) at the N-terminus of BocN-carbo beta,alpha- and beta,beta-dipeptides. PMID- 16220505 TI - Group sequential, sample size re-estimation and two-stage adaptive designs in clinical trials: a comparison. PMID- 16220506 TI - An evaluation of Bayesian designs for dose-escalation studies in healthy volunteers. AB - In this paper, Bayesian decision procedures previously proposed for dose escalation studies in healthy volunteers are reviewed and evaluated. Modifications are made to the expression of the prior distribution in order to make the procedure simpler to implement and a more relevant criterion for optimality is introduced. The results of an extensive simulation exercise to establish the properties of the procedure and to aid choice between designs are summarized, and the way in which readers can use simulation to choose a design for their own trials is described. The influence of the value of the within subject correlation on the procedure is investigated and the use of a simple prior to reflect uncertainty about the correlation is explored. PMID- 16220507 TI - Decomposition of life expectancy and expected life-years lost by disease. AB - Life expectancy is commonly used to summarize the life-time mortality experience of a population. Differences in life expectancy are well-known across different levels of socioeconomic status such as income and education. A recent simulation study of potential life-years lost has shown the effects that major diseases contribute to differences in life expectancy at birth. We propose a general methodology to decompose life expectancy and expected life-years lost by disease in order to determine the contribution of diseases to differences in life expectancy at each given age. We show that the estimates for the life expectancy, expected life-years lost and their variances at each age can be computed backward recursively from an old age. The difference in life expectancy between groups will be shown to include contributions from diseases and life-year differences which occur after an old age cut-off beyond which the contribution of diseases cannot be easily determined. Diseases will be grouped into 14 major disease categories. Data from the National Longitudinal Mortality Study will be used for demonstration purposes. PMID- 16220508 TI - Additive models for geo-referenced failure time data. AB - Asthma researchers have found some evidence that geographical variations in susceptibility to asthma could reflect the effect of community level factors such as exposure to violence. Our methodology was motivated by a study of age at onset of asthma among children of inner-city neighbourhoods in East Boston. Cox's proportional hazards model was not appropriate since there was not enough information about the nature of geographical variations so as to impose a parametric relationship. In addition, some of the known risk factors were believed to have non-linear log-hazard ratios. We extend the geoadditive models of Kamman and Wand to the case where the outcome measure is a possibly censored time to event. We reduce the problem to one of fitting a Poisson mixed model by using Poisson approximations in conjunction with a mixed model formulation of generalized additive modelling. Our method allows for low-rank additive modelling, provides likelihood-based estimation of all parameters including the amount of smoothing and can be implemented using standard software. We illustrate our method on the East Boston data. PMID- 16220509 TI - A robust approach to t linear mixed models applied to multiple sclerosis data. AB - We discuss a robust extension of linear mixed models based on the multivariate t distribution. Since longitudinal data are successively collected over time and typically tend to be auto-correlated, we employ a parsimonious first-order autoregressive dependence structure for the within-subject errors. A score test statistic for testing the existence of autocorrelation among the within-subject errors is derived. Moreover, we develop an explicit scoring procedure for the maximum likelihood estimation with standard errors as a by-product. The technique for predicting future responses of a subject given past measurements is also investigated. Results are illustrated with real data from a multiple sclerosis clinical trial. PMID- 16220510 TI - Constructing intervals for the intracluster correlation coefficient using Bayesian modelling, and application in cluster randomized trials. AB - Studies in health research are commonly carried out in clustered settings, where the individual response data are correlated within clusters. Estimation and modelling of the extent of between-cluster variation contributes to understanding of the current study and to design of future studies. It is common to express between-cluster variation as an intracluster correlation coefficient (ICC), since this measure is directly comparable across outcomes. ICCs are generally reported unaccompanied by confidence intervals. In this paper, we describe a Bayesian modelling approach to interval estimation of the ICC. The flexibility of this framework allows useful extensions which are not easily available in existing methods, for example assumptions other than Normality for continuous outcome data, adjustment for individual-level covariates and simultaneous interval estimation of several ICCs. There is also the opportunity to incorporate prior beliefs on likely values of the ICC. The methods are exemplified using data from a cluster randomized trial. PMID- 16220511 TI - Bayesian evidence synthesis to extrapolate survival estimates in cost effectiveness studies. AB - This paper is concerned with survival extrapolation that represents an integral part of cost-effectiveness analysis. In the absence of long-term survival estimates from randomized clinical trials or meta-analysis we show how age-sex matched U.K. population data can additionally be used to estimate survival patterns. We adopt a Bayesian approach and we synthesize evidence from different sources such as patient registries, U.K. population statistics and meta-analyses. We also present methodology for Bayesian analysis of the additive hazards model and we show how to apply the techniques using freely available software. The methods are illustrated using data from a cohort of cardiac arrhythmia patients. PMID- 16220512 TI - Comparison of variance estimation approaches in a two-state Markov model for longitudinal data with misclassification. AB - We examine the behaviour of the variance-covariance parameter estimates in an alternating binary Markov model with misclassification. Transition probabilities specify the state transitions for a process that is not directly observable. The state of an observable process, which may not correctly classify the state of the unobservable process, is obtained at discrete time points. Misclassification probabilities capture the two types of classification errors. Variance components of the estimated transition parameters are calculated with three estimation procedures: observed information, jackknife, and bootstrap techniques. Simulation studies are used to compare variance estimates and reveal the effect of misclassification on transition parameter estimation. The three approaches generally provide similar variance estimates for large samples and moderate misclassification. In these situations, the resampling methods are reasonable alternatives when programming partial derivatives is not appealing. With smaller chains or higher misclassification probabilities, the bootstrap method appears to be the best choice. PMID- 16220513 TI - A powerful method of combining measures of association and Hardy-Weinberg disequilibrium for fine-mapping in case-control studies. AB - We present a new method for fine-mapping a disease susceptibility locus using a case-control design. The new method, termed the 'weighted average (WA) statistic', averages the Cochran-Armitage (CA) trend test statistic and the difference between the Hardy Weinberg disequilibrium test statistics (the HWD trend) for cases and controls. The main features of the WA statistic are that it mitigates against the weaknesses, and maintains the strong points, of both the CA trend test and the HWD trend test. To allow for the extra variance induced by population structure and cryptic relatedness, the WA statistic can be adjusted for variance inflation. Based on the results of a simulation study, when there is no population structure the WA test statistic shows good performance under a variety of genetic disease models. When there is population structure, the adjusted WA statistic maintains the correct probability of type I error. Under all genetic disease models investigated, the adjusted WA statistic has better power than the adjusted CA trend test, the HWD trend test or the product of the adjusted CA trend test and the HWD trend test statistics. PMID- 16220515 TI - Multiple imputation techniques in small sample clinical trials. AB - Clinical trials allow researchers to draw conclusions about the effectiveness of a treatment. However, the statistical analysis used to draw these conclusions will inevitably be complicated by the common problem of attrition. Resorting to ad hoc methods such as case deletion or mean imputation can lead to biased results, especially if the amount of missing data is high. Multiple imputation, on the other hand, provides the researcher with an approximate solution that can be generalized to a number of different data sets and statistical problems. Multiple imputation is known to be statistically valid when n is large. However, questions still remain about the validity of multiple imputation for small samples in clinical trials. In this paper we investigate the small-sample performance of several multiple imputation methods, as well as the last observation carried forward method. PMID- 16220514 TI - Diagnosis using predictive probabilities without cut-offs. AB - Standard diagnostic test procedures involve dichotomization of serologic test results. The critical value or cut-off is determined to optimize a trade off between sensitivity and specificity of the resulting test. When sampled units from a population are tested, they are allocated as either infected or not according to the test outcome. Units with values high above the cut-off are treated the same as units with values just barely above the cut-off, and similarly for values below the cut-off. There is an inherent information loss in dichotomization. We thus develop a diagnostic screening method based on data that are not dichotomized within the Bayesian paradigm. Our method determines the predictive probability of infection for each individual in a sample based on having observed a specific serologic test result and provides inferences about the prevalence of infection in the population sampled. Our fully Bayesian method is briefly compared with a previously developed frequentist method. We illustrate the methodology with serologic data that have been previously analysed in the veterinary literature, and also discuss applications to screening for disease in humans. The method applies more generally to a variation of the classic parametric 2-population discriminant analysis problem. Here, in addition to training data, additional units are sampled and the goal is to determine their population status, and the prevalence(s) of the subpopulation(s) from which they were sampled. PMID- 16220516 TI - Generalized gamma frailty model. AB - In this article, we present a frailty model using the generalized gamma distribution as the frailty distribution. It is a power generalization of the popular gamma frailty model. It also includes other frailty models such as the lognormal and Weibull frailty models as special cases. The flexibility of this frailty distribution makes it possible to detect a complex frailty distribution structure which may otherwise be missed. Due to the intractable integrals in the likelihood function and its derivatives, we propose to approximate the integrals either by Monte Carlo simulation or by a quadrature method and then determine the maximum likelihood estimates of the parameters in the model. We explore the properties of the proposed frailty model and the computation method through a simulation study. The study shows that the proposed model can potentially reduce errors in the estimation, and that it provides a viable alternative for correlated data. The merits of proposed model are demonstrated in analysing the effects of sublingual nitroglycerin and oral isosorbide dinitrate on angina pectoris of coronary heart disease patients based on the data set in Danahy et al. (sustained hemodynamic and antianginal effect of high dose oral isosorbide dinitrate. Circulation 1977; 55:381-387). PMID- 16220517 TI - The reassessment of trial perspectives from interim data--a critical view. AB - If an interim analysis is performed during a trial it is tempting to determine the conditional power to reach a rejection in the trial given the observed results in the interim analysis. Since the true effect size is unknown the conditional power may be calculated by using the effect size, which the study has been powered for in the planning phase or by using an interim estimate of the true size (or a combination of both). In either case the conditional power is a random variable and its density is investigated depending on the analysis time and the true effect size. Under the null hypothesis, in early interim analyses after a small proportion of sample units, the conditional power typically will be close to the overall power when the effect size from the planning stage is used for calculation. In this case the majority of observations must still be made and the small first-stage sample in general will be dominated by the hypothetical second-stage chance based on the wrong parameter value. It is shown that the conditional power in moderately underpowered studies can have a distribution symmetric around 0.5. When using the interim estimate for calculating the conditional power the density in general will be u-shaped. The impact of using conditional power to reassess the sample size using flexible two-stage combination tests is shown for a specific example in terms of overall power and average sample size as compared to the corresponding group sequential design. For small true effect sizes mid-trial sample size recalculation based on an interim estimate may lead to an overly large price to be paid in average sample size in relation to the gain in overall power. Finally, the problem is discussed in terms of estimating the true conditional power. PMID- 16220518 TI - Tutorial in biostatistics: the self-controlled case series method. AB - The self-controlled case series method was developed to investigate associations between acute outcomes and transient exposures, using only data on cases, that is, on individuals who have experienced the outcome of interest. Inference is within individuals, and hence fixed covariates effects are implicitly controlled for within a proportional incidence framework. We describe the origins, assumptions, limitations, and uses of the method. The rationale for the model and the derivation of the likelihood are explained in detail using a worked example on vaccine safety. Code for fitting the model in the statistical package STATA is described. Two further vaccine safety data sets are used to illustrate a range of modelling issues and extensions of the basic model. Some brief pointers on the design of case series studies are provided. The data sets, STATA code, and further implementation details in SAS, GENSTAT and GLIM are available from an associated website. PMID- 16220519 TI - Estimating transitions between symptom severity states over time in schizophrenia: a Bayesian meta-analytic approach. AB - We obtain the posterior predictive distribution of transition probabilities between symptom severity states over time for patients with schizophrenia by (i) employing a Bayesian meta-analysis of published clinical trials and observational studies to estimate the posterior distribution of parameters that guide changes in Positive and Negative Syndrome Scale (PANSS) scores over time and under the influence of various drugs and (ii) by propagating the variability from the posterior distributions of the parameters through a micro-simulation model that is formulated based on schizophrenia progression. Results show detailed differences among haloperidol, risperidone and olanzapine in controlling various levels of severities of positive, negative and joint symptoms over time. For example, risperidone seems best in controlling severe positive symptoms while olanzapine is the worst in that during the first quarter of drug treatment; however, olanzapine seems to be best in controlling severe negative symptoms across all four quarters of treatment while haloperidol is the worst in this regard. These details may further serve to better estimate quality of life of patients and aid in resource utilization decisions in treating schizophrenic patients. In addition, consistent estimation of uncertainty in the time-profile parameters also has important implications for the practice of cost-effectiveness analysis and for future resource allocation policies in schizophrenia treatment. PMID- 16220520 TI - Structured correlation in models for clustered data. AB - Correlation is always a concern in the analysis of clustered data. One area of interest is to develop a general correlation modelling approach for high dimensional data with unbalanced hierarchical and heterogeneous data structures, e.g. multilevel data. Commonly used correlation structures might have limitation for such situations. In this paper, we propose two extensions, multiblock and multilayer correlations. These methods are very flexible in modelling correlation and can be incorporated in many multivariate approaches, while the major discussion focuses on the applications under the generalized estimating equations (GEE) methods. The approaches are especially useful in GEE when each cluster is large and complex but the number of clusters is small. If an incorrect correlation is applied to such data, the results are less efficient. Multiblock and multilayer correlations extend GEE methods to model complicated multilevel data with arbitrary number of levels and cluster size. The extended estimating equation for correlation parameters has an orthogonal property, and the computation is very efficient. A simulation study compares the conventional methods versus the proposed methods, and it shows the gain in relative efficiency and the flexibility in modelling various structures. PMID- 16220521 TI - Model comparison of generalized linear mixed models. AB - Generalized linear mixed models (GLMMs) have been widely appreciated in biological and medical research. Maximum likelihood estimation has received a great deal of attention. Comparatively, not much has been done on model comparison or hypotheses testing. In this article, we propose a path sampling procedure to compute the observed-data log-likelihood function, so that the Bayesian information criterion (BIC) can be applied to model comparison or hypothesis testing. Advantages of the proposed path sampling procedure are discussed. Two medical data sets are analysed for providing illustrative examples of the proposed methodology. PMID- 16220522 TI - A multi-state model for joint modelling of terminal and non-terminal events with application to Whitehall II. AB - Serious coronary heart disease (CHD) is a primary outcome in the Whitehall II study, a large epidemiological study of British civil servants. Both fatal (F) and non-fatal (NF) CHD events are of interest and while essentially complete information is available on F events, the observation of NF events is subject to potentially informative censoring. A multi-state model with an unobserved state is introduced for the joint modelling of F and NF events. Two model-based assumptions ensure identifiability of the model and a parameter is introduced to allow sensitivity analyses concerning the assumption linked to informative censoring. Weibull transition rates, which include dependence on explanatory variables, are used in the analysis of Whitehall II data with a particular focus on the relationship between civil service grade and CHD events. PMID- 16220523 TI - Structure elucidation and reaction monitoring of synthetic intermediates using an automated accurate mass GC-TOF mass spectrometer in drug discovery. PMID- 16220524 TI - Efficient group sequential designs when there are several effect sizes under consideration. AB - We consider the construction of efficient group sequential designs where the goal is a low expected sample size not only at the null hypothesis and the alternative (taken to be the minimal clinically meaningful effect size), but also at more optimistic anticipated effect sizes. Pre-specified Type I error rate and power requirements can be achieved both by standard group sequential tests and by more recently proposed adaptive procedures. We investigate four nested classes of designs: (A) group sequential tests with equal group sizes and stopping boundaries determined by a monomial error spending function (the 'rho-family'); (B) as A but the initial group size is allowed to be different from the others; (C) group sequential tests with arbitrary group sizes and arbitrary boundaries, fixed in advance; (D) adaptive tests-as C but at each analysis, future group sizes and critical values are updated depending on the current value of the test statistic. By examining the performance of optimal procedures within each class, we conclude that class B provides simple and efficient designs with efficiency close to that of the more complex designs of classes C and D. We provide tables and figures illustrating the performances of optimal designs within each class and defining the optimal procedures of classes A and B. PMID- 16220525 TI - Theoretical study of the gas-phase structures of sodiated and cesiated leucine and isoleucine: zwitterionic structure disfavored in kinetic method experiments. AB - The most stable charge-solvated (CS) and zwitterionic (ZW) structures of sodiated and cesiated leucine and isoleucine were studied by density functional theory methods. According to the Boltzmann distribution in gas phase, both forms of LeuNa+ and IleNa+ exist, but in LeuCs+ and IleCs+, the ZW forms are dominant. Results for the sodiated compounds are consistent with the relationship found between decrease in relative stability of CS versus ZW form and aliphatic amino acid side chain length. The observed degeneracy in energy for IleNa+ conformers is at odds with kinetic method results. Additional calculations showed that kinetic method structural determinations for IleNa+ do not reflect relative order of populations in the lowest energy conformers. Since complexation of cationized amino acids into ion-bound dimers disfavors ZW structure by approximately 8 kJ mol(-1), it is suggested that for energy close conformers of sodium-cationized amino acids, the kinetic method may not be reliable for structural determinations. PMID- 16220526 TI - A novel on-line solid-phase extraction approach integrated with a monolithic column and tandem mass spectrometry for direct plasma analysis of multiple drugs and metabolites. AB - An on-line solid-phase extraction liquid chromatography/tandem mass spectrometry (SPE LC/MS/MS) assay using a newly developed SPE column and a monolithic column was developed and validated for direct analysis of plasma samples containing multiple analytes. This assay was developed in an effort to increase bioanalysis throughput and reduce the complexity of on-line SPE LC/MS/MS systems. A simple column-switching configuration that requires only one six-port valve and one HPLC pumping system was employed for on-line plasma sample preparation and subsequent gradient chromatographic separation. The resulting analytical method couples the desired sensitivity with ease of use. The method was found to perform satisfactorily for direct plasma analysis with respect to assay linearity, specificity, sensitivity, precision, accuracy, carryover, and short-term stability of an eight-analyte mixture in plasma. A gradient LC condition was applied to separate the eight analytes that cannot be distinctly differentiated by MS/MS. With a run time for every injection of 2.8 min, a minimum of 300 direct plasma injections were made on one on-line SPE column without noticeable changes in system performance. Due to the ruggedness and simplicity of this system, generic methods can be easily developed and applied to analyze a wide variety of compounds in a high-throughput manner without laborious off-line sample preparation. PMID- 16220527 TI - A novel dual-isotope labelling method for distinguishing between soil sources of N2O. AB - We present a novel 18O-15N-enrichment method for the distinction between nitrous oxide (N2O) from nitrification, nitrifier denitrification and denitrification based on a method with single- and double-15N-labelled ammonium nitrate. We added a new treatment with 18O-labelled water to quantify N2O from nitrifier denitrification. The theory behind this is that ammonia oxidisers use oxygen (O2) from soil air for the oxidation of ammonia (NH3), but use H2O for the oxidation of the resulting hydroxylamine (NH2OH) to nitrite (NO2-). Thus, N2O from nitrification would therefore be expected to reflect the 18O signature of soil O2, whereas the 18O signature of N2O from nitrifier denitrification would reflect that of both soil O2 and H2O. It was assumed that (a) there would be no preferential removal of 18O or 16O during nitrifier denitrification or denitrification, (b) the 18O signature of the applied 18O-labelled water would remain constant over the experimental period, and (c) any O exchange between H(2)18O and NO3- would be negligible under the chosen experimental conditions. These assumptions were tested and validated for a silt loam soil at 50% water filled pore space (WFPS) following application of 400 mg N kg-1 dry soil. We compared the results of our new method with those of a conventional inhibition method using 0.02% v/v acetylene (C2H2) and 80% v/v O2 in helium. Both the 18O 15N-enrichment and inhibitor methods identified nitrifier denitrification to be a major source of N2O, accounting for 44 and 40%, respectively, of N2O production over 24 h. However, compared to our 18O-15N-method, the inhibitor method overestimated the contribution from nitrification at the expense of denitrification, probably due to incomplete inhibition of nitrifier denitrification and denitrification by large concentrations of O2 and a negative effect of C2H2 on denitrification. We consider our new 18O-15N-enrichment method to be more reliable than the use of inhibitors; it enables the distinction between more soil sources of N2O than was previously possible and has provided the first direct evidence of the significance of nitrifier denitrification as a source of N2O in fertilised arable soil. PMID- 16220529 TI - Proteomic profiling of cholangiocarcinoma cell line treated with pomiferin from Derris malaccensis. AB - Pomiferin, a prenylated isoflavonoid from Derris malaccensis with strong anti fungal and anti-oxidant activities, showed cytotoxic activity towards human cholangiocarcinoma cells (HuCCA-1), with IC(50) of 0.9 microg/mL. Pomiferin caused apoptosis, detectable by DNA fragmentation. Two-dimensional PAGE showed increased expression of 12 proteins, namely glucose-regulated protein 75 (grp 75), calcyclin (S100A6), degraded cytokeratin 19, ATP synthase D, ribosomal protein P0, degraded cytokeratin 18 (two spots pI/MW 6.03/29.9 and pI/MW 4.66/21.5), cofilin, annexin A1, triose phosphate isomerase, peroxiredoxin-1, calgizzarin, and profilin. In contrast, cytokeratins (CK) 7, 18 and 19 were down regulated, and were shown by 1-DE immunodetection to be degraded. PMID- 16220530 TI - An enhanced method for peptides sequencing by N-terminal derivatization and MS. AB - An improved method for peptide sequencing based on acetylation/deuteroacetylation in conjunction with ESI MS is introduced. Derivatization with a 1:1 mixture of acetic anhydride and deuterated acetic anhydride incorporates a stable isotope label into the analyzed molecule. This approach has been initially applied to FAB. Using MS/MS, the technique provides a fast, highly sensitive and reliable determination of the primary structure of unknown peptides. This procedure labels N-terminal fragments formed during MS/MS analysis, resulting in a simplification and faster interpretation of the spectra. The performance of the method has been tested with several synthetic peptides and applied to an efficient sequencing of the peptide map, using a nano-scale LC coupled on-line to a tandem mass spectrometer. PMID- 16220531 TI - Proteomic tracking of serum protein isoforms as screening biomarkers of ovarian cancer. AB - Epithelial ovarian cancer is the fourth leading cause of cancer death among women. Due to the asymptomatic nature and poor survival characteristic of the disease, screening for specific biomarkers for ovarian cancer is a major health priority. Differentially expressed proteins in the serum of ovarian cancer patients have the potential to be used as cancer-specific biomarkers. In this study, proteomic methods were used to screen 24 serum samples from women with high-grade ovarian cancer and compared to a control group of 11 healthy women. Affigel-Blue treated serum samples were processed either by linear (pH 4-7) or narrow range (pH 5.5-6.7) IEF strips for the first dimension. Proteins separated in first dimension were resolved by 8-16% gradient SDS-PAGE. Protein spots were visualized by SYPRO Ruby staining, imaged by FX-imager and compared and analyzed by PDQuest software. Twenty-two protein spots were consistently differentially expressed between normal and ovarian cancer patients by resolving proteins in a linear pH strip of 4-7 for the first dimension. Six of the protein spots, significantly up-regulated in grade 3 ovarian cancer patients (p < 0.05), were identified by MALDI-TOF MS and Western blotting as the isoforms of haptoglobin precursor. When serum proteins were resolved on narrow pH range strips (5.5-6.7), 23 spots were consistently differentially expressed between normal and grade 3 ovarian cancer patients. Of these, 4 protein spots significantly down regulated in grade 3 ovarian cancer patients (p < 0.05) were identified by MALDI-TOF MS and Western blotting, as isoforms of transferrin precursor. Increased expression of serum haptoglobin and transferrin was also identified in peritoneal tumor fluid obtained from women diagnosed with grade 2/3 ovarian cancer (n = 7). Changes in the expression of haptoglobin and transferrin in the serum of women with different pathological grades of ovarian cancer was examined by one-dimensional Western blotting method. Serum samples collected from women suffering from benign, borderline, grade 1, grade 2 and grade 3 cancer (n = 4 for haptoglobin and n = 5 for transferrin in each group) were analyzed and compared to the serum of normal healthy women. The mean serum haptoglobin expression in grade 3 ovarian cancer patients was fourfold higher than in the control subjects (p < 0.05). On the other hand, transferrin expression in grade 3 ovarian cancer patients was decreased by twofold than in normal healthy women (p < 0.05). Haptoglobin expression in the serum of cancer patients (n = 7) decreased following chemotherapy (six cycles of taxol/carboplatin). Concomitant with the decrease of haptoglobin, transferrin expression remained constant in four patients, but increased in three out of seven patients included in the study. Changes in serum expression of haptoglobin correlated with the change of CA 125 levels before and after chemotherapy. In conclusion, proteomic profiling of differentially expressed proteins in the sera of normal women compared to women with ovarian cancer can greatly facilitate the discovery of a panel of biomarkers that may aid in the detection of ovarian cancer with greater specificity. PMID- 16220532 TI - Proteome of endothelial cell-derived procoagulant microparticles. AB - Microparticles (MP) are small membrane vesicles that are released from cells upon activation or during apoptosis. Cellular MP in body fluids constitute a heterogeneous population, differing in cellular origin, numbers, size, antigenic composition and functional properties. MP support coagulation by exposure of tissue factor (TF), the initiator of coagulation in vivo. Moreover, MP may transfer bioactive molecules to other cells, thereby stimulating them to produce cytokines, cell-adhesion molecules, growth factors and TF, and modulate endothelial functions. However, a comprehensive characterization of the antigenic composition of MP has been poorly defined. This study describes the protein composition of endothelial cell (EC)-derived MP (EMP) using a proteomic approach. MS analysis indicated the presence of newly described protein such as metabolic enzymes, proteins involved in adhesion and fusion processes, members of protein folding event, cytoskeleton associated proteins and nucleosome. In conclusion, circulating EMP behave as an actual storage pool, able to disseminate blood-borne TF activity and other bioactive effectors, as confirmed by our experiments showing an increased procoagulant activity of EC exposed to EMP. PMID- 16220533 TI - Proteomic analysis of the excretory-secretory proteins of the Trichinella spiralis L1 larva, a nematode parasite of skeletal muscle. AB - Trichinella spiralis is an intracellular nematode parasite of mammalian skeletal muscle. Infection of the muscle cell leads to the formation of a host-parasite complex that results in profound alterations to the host cell and a re-alignment of muscle-specific gene expression. The role of parasite excretory-secretory (ES) proteins in mediating these effects is currently unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, a global proteomics approach was used to analyse the ES proteins from T. spiralis muscle larvae. Following 2-DE of ES proteins,MALDI-TOF-MS and LC-MS/MS were used to identify the peptide spots. Specific Trichinella EST databases were assembled and used to analyse the data. Despite the current absence of a Trichinella genome-sequencing project, 43 out of 52 protein spots analysed were identified and included the major secreted glycoproteins. Other novel proteins were identified from matches with sequences in the T. spiralis database. Our results demonstrate the value of proteomics as a tool for the identification of Trichinella ES proteins and in the study of the molecular mechanism underpinning the formation of the host-parasite complex during Trichinella infections. PMID- 16220534 TI - Proteomics-based consensus prediction of protein retention in a bacterial membrane. AB - The availability of complete bacterial genome sequences allows proteome-wide predictions of exported proteins that are potentially retained in the cytoplasmic membranes of the corresponding organisms. In practice, however, major problems are encountered with the computer-assisted distinction between (Sec-type) signal peptides that direct exported proteins into the growth medium and lipoprotein signal peptides or amino-terminal membrane anchors that cause protein retention in the membrane. In the present studies, which were aimed at improving methods to predict protein retention in the bacterial cytoplasmic membrane, we have compared sets of membrane-attached and extracellular proteins of Bacillus subtilis that were recently identified through proteomics approaches. The results showed that three classes of membrane-attached proteins can be distinguished. Two classes include 43 lipoproteins and 48 proteins with an amino-terminal transmembrane segment, respectively. Remarkably, a third class includes 31 proteins that remain membrane-retained despite the presence of typical Sec-type signal peptides with consensus signal peptidase recognition sites. This unprecedented finding indicates that unknown mechanisms are involved in membrane retention of this class of proteins. A further novelty is a consensus sequence indicative for release of certain lipoproteins from the membrane by proteolytic shaving. Finally, using non-overlapping sets of secreted and membrane-retained proteins, the accuracy of different signal peptide prediction algorithms was assessed. Accuracy for the prediction of protein retention in the membrane was increased to 82% using a majority-vote approach. Our findings provide important leads for future identification of surface proteins from pathogenic bacteria, which are attractive candidate infection markers and potential targets for drugs or vaccines. PMID- 16220535 TI - Advantages and limitations of clear-native PAGE. AB - Clear-native PAGE (CN-PAGE) separates acidic water-soluble and membrane proteins (pI < 7) in an acrylamide gradient gel, and usually has lower resolution than blue-native PAGE (BN-PAGE). The migration distance depends on the protein intrinsic charge, and on the pore size of the gradient gel. This complicates estimation of native masses and oligomerization states when compared to BN-PAGE, which uses negatively charged protein-bound Coomassie-dye to impose a charge shift on the proteins. Therefore, BN-PAGE rather than CN-PAGE is commonly used for standard analyses. However, CN-PAGE offers advantages whenever Coomassie-dye interferes with techniques required to further analyze the native complexes, e.g., determination of catalytic activities, as shown here for mitochondrial ATP synthase, or efficient microscale separation of membrane protein complexes for fluorescence resonance energy transfer (FRET) analyses. CN-PAGE is milder than BN PAGE. Especially the combination of digitonin and CN-PAGE can retain labile supramolecular assemblies of membrane protein complexes that are dissociated under the conditions of BN-PAGE. Enzymatically active oligomeric states of mitochondrial ATP synthase previously not detected using BN-PAGE were identified by CN-PAGE. PMID- 16220536 TI - Properties of murine (CD8+)CD27- T cells. AB - In humans, loss of CD27 expression is associated with the stable acquisition of effector functions by CD8+ T cells. We found that murine (CD8+)CD27- T cells were confined to the primed CD62L(dull/-)CD44(bright)CCR7- T cell population. (CD8+)CD27- T cells were absent from lymph nodes but could be found in blood, spleen and in non-lymphoid organs such as lung and liver. Late after primary influenza virus infection, low percentages of antigen-specific CD27- cells emerged in the lung and spleen. After recovery from secondary influenza virus infection, high percentages of influenza-specific CD27- T cells were found in the lung and the loss of CD27 on lung CD8+ T cells coincided with high granzyme B expression. After murine cytomegalovirus infection, loss of CD27 expression on virus-specific CD8+ T cell populations was sustained and especially marked in liver and lung. We suggest that in mice, CD27 is lost from CD8+ T cells only after repetitive antigenic stimulation. Moreover, the high expression of both granzyme B and perforin in the CD27- T cells suggests that the lack of CD27 on murine CD8+ T cells can be used to identify memory T cells with expression of cytotoxic effector molecules. PMID- 16220537 TI - Novel phenotypes and migratory properties distinguish memory CD4 T cell subsets in lymphoid and lung tissue. AB - Memory T cells are heterogeneous in expression of lymph node homing receptors, delineating "central-memory" (TCM, CD62Lhi/CCR7+) and "effector-memory" (TEM, CD62Llo/CCR7-) subsets that migrate to lymphoid and non-lymphoid tissues, respectively. It is not known how these subsets arise or how homing receptor expression and tissue origin determine their functional and migratory properties. Here, we investigated the role of CD62L expression in the generation, function, distribution and migration of heterogeneous memory CD4 T cells specific for influenza hemagglutinin (HA). We found that CD62Lhi and CD62Llo memory subsets are generated independent of CD62L expression by the activated precursor, and both subsets distribute into spleen and lung. Functionally, spleen- and lung derived CD62L memory subsets produce effector cytokines at similar kinetics but differ strikingly in cell surface phenotype and migration: the CD62Llo memory subset expresses a classic memory phenotype (CD45RBlo/CD44hi/CD11a(hi)), while the CD62Lhi subset expresses an unconventional phenotype (CD45RBhi/CD44int/CD11a(int)), defining a new polyclonal memory subset. The CD62Lhi subset also trafficked more efficiently than CD62Llo cells into lymph nodes; however, only lung but not spleen CD62Llo memory T cells homed to lung. Our results reveal novel phenotypic heterogeneity of memory CD4 T cells co segregating with CD62L expression and tissue-specific tropism of non-lymphoid memory CD4 T cells. PMID- 16220538 TI - Identification of genes involved in the initiation of human Th1 or Th2 cell commitment. AB - The differentiation of naive T helper (Th) cells is induced by TCR activation and IL-12/STAT4 or IL-4/STAT6 signaling pathways, forming Th1 and Th2 cells, respectively. In this study, oligonucleotide arrays were used to identify genes regulated during the initiation of human Th1 and Th2 cell differentiation at 2 and 6 h in presence or absence of immunosuppressive TGF-beta. As a result the immediate targets of IL-12, IL-4 and TGF-beta were identified. The effects of IL 12 at this early stage were minimal and consistent with the known kinetics of IL 12Rbeta2 expression. IL-4, however, was observed to rapidly regulate 63 genes, 26 of which were differentially expressed at both the 2- and 6-h time points. Of these IL-4 regulated genes, one-third have previously been observed to display expression changes in the later phases of the polarization process. Similarly to the key regulators, TBX21 and GATA3, the transcription factors SATB1, TCF7 and BCL6 were differentially regulated at the protein level during early Th1 and Th2 cell polarization. Moreover, the developing Th1 and Th2 cells were demonstrated to be responsive to the immunosuppressive TGF-beta and IL-10. In this study, a panel of novel factors that may be important regulators of the differentiation process was identified. PMID- 16220539 TI - IL-12 up-regulates T-bet independently of IFN-gamma in human CD4+ T cells. AB - T-bet is an important Th1 driving transcription factor regulated by IFN gamma/STAT1 pathway. T-bet turns on IFN-gamma transcription in CD4+ T cells and T bet-deficient cells fail to differentiate to Th1 direction. Previous reports have characterized function of T-bet mainly in murine cells and very little is known about its functions in human cells. Here, we studied T-bet expression kinetics in parallel with GATA3 during Th1/Th2 polarization. We demonstrate that in addition to CD3/CD28 activation, cytokines IL-12 and IFN-alpha in the presence of neutralizing anti-IFN-gamma enhanced T-bet mRNA and protein expression in human CD4+ cells. T-bet is known to be a potent inducer of IFN-gamma. Even though IFN gamma and IL-12 stimulation induced similar levels of T-bet protein in human CD4+ cells, IFN-gamma-treated cells produced considerably less IFN-gamma than cells treated with IL-12. Therefore, high T-bet protein expression does not necessarily correlate with IFN-gamma production. In addition, we show that the immunosuppressive cytokine TGF-beta inhibits T-bet and GATA3 protein expression only if it is present prior to primary T cell activation and is maintained in the cultures during the early polarization of Th1/Th2 cells. In conclusion, we report new insights into the cytokine regulation of T-bet in human CD4+ T cells. PMID- 16220540 TI - Activation of the Nrf2/antioxidant response pathway increases IL-8 expression. AB - Oxidant stress can initiate or enhance inflammatory responses during tissue injury, possibly through activation of redox-sensitive chemokines. Because the transcription factor Nrf2 (NF-E2-related factor 2) is responsive to oxidative stress, and induces expression of cytoprotective and antioxidant genes that attenuate tissue injury, we postulated that Nrf2 may also regulate chemokine expression. To test this hypothesis, Nrf2 expression was directly increased in primary human kidney mesangial cells and aortic endothelial cells, or cell lines with an adenoviral construct, and the effects on the pro-inflammatory chemokine interleukin-8 (IL-8) were assessed. Nrf2 expression significantly increased IL-8 mRNA levels and protein secretion. Nrf2 caused only a weak induction of IL-8 transcription, but significantly increased the half-life of IL-8 mRNA. These data demonstrate that activation of the Nrf2/antioxidant response pathway induces expression of IL-8. The dominant mechanism of Nrf2-mediated IL-8 induction is through mRNA stabilization. Considering the evidence that Nrf2 activation is mainly cytoprotective, these observations raise the possibility that under certain circumstances IL-8 may serve an anti-inflammatory role and thereby contribute to the resolution of tissue injury. PMID- 16220542 TI - IFN-gamma-producing human T cells directly induce osteoclastogenesis from human monocytes via the expression of RANKL. AB - The current study explored our hypothesis that IFN-gamma-producing human T cells inhibit human osteoclast formation. Activated T cells derived from human PBMC were divided into IFN-gamma-producing T cells (IFN-gamma(+) T cells) and IFN gamma-non-producing T cells (IFN-gamma(-) T cells). IFN-gamma(+) T cells were cultured with human monocytes in the presence of macrophage-CSF alone. The concentration of soluble receptor activator of NF-kappaB ligand (RANKL) and IFN gamma, and the amount of membrane type RANKL expressed on T cells, were measured by ELISA. In the patients with early rheumatoid arthritis (RA) treated with non steroidal anti-inflammatory drugs alone, CD4+ T cells expressing both IFN-gamma and RANKL were detected by flow cytometry. Surprisingly, IFN-gamma(+) T cells, but not IFN-gamma(-) T cells, induced osteoclastogenesis from monocytes, which was completely inhibited by adding osteoprotegerin and increased by adding anti IFN-gamma antibodies. The levels of both soluble and membrane type RANKL were elevated in IFN-gamma(+) T cells. The ratio of CD4+ T cells expressing both IFN gamma and RANKL in total CD4+ T cells from PBMC was elevated in RA patients. Contrary to our hypothesis, IFN-gamma(+) human T cells induced osteoclastogenesis through the expression of RANKL, suggesting that Th1 cells play a direct role in bone resorption in Th1 dominant diseases such as RA. PMID- 16220541 TI - Peptide-stimulated DO11.10 T cells divide well but accumulate poorly in the absence of TLR agonist treatment. AB - Immunological adjuvants increase the clonal burst size of antigen-specific T cell populations by mechanisms that remain incompletely understood. Using the DO11.10 adoptive transfer system to study peptide-stimulated T cell responses, we found that TLR agonist treatment increased the extent of cellular division undergone by responding T cells, but not by enough to explain the net increases in T cell yield that were achieved. Two novel analyses involving CFSE dye dilution analysis were used to characterize the shortfall, both of which were consistent with the idea that DO11.10 T cells are frequently lost during proliferation unless TLR agonists are present. T cell loss during clonal expansion was correlated with decreased levels of Bcl-2, but TLR agonists did not appear to afford protection by restoring levels of Bcl-2 or of cell surface IL-7Ralpha chain expression. TLR mediated protection also failed to correlate with increased expression of Bcl-x or decreased expression of pro-apoptotic Bim. Our findings suggest that DO11.10 T cells stimulated by antigenic peptide in vivo divide well, but fail to accumulate efficiently unless TLR agonists are present. PMID- 16220543 TI - Cloned endangered species takin (Budorcas taxicolor) by inter-species nuclear transfer and comparison of the blastocyst development with yak (Bos grunniens) and bovine. AB - Interspecies cloning might be used as an effective method to conserve endangered species and to support the study of nuclear-cytoplasm interaction. In this study, we describe the development of takin-bovine embryos in vitro produced by fusing takin ear fibroblasts with enucleated bovine oocytes and examine the fate of mitochondrial DNA in these embryos. We also compare the blastocyst development of takin-bovine embryos with yak-bovine and bovine-bovine embryos and compare the cell numbers of the blastocyst. Our results indicate that: (1) takin-bovine cloned embryos can develop to the blastocyst stage in vitro (5%), (2) blastocyst mitochondria DNA are derived primarily from bovine oocytes in spite of a little takin donor cell mitochondrial DNA, (3) using the same cloned protocol, development efficiency is significantly different between bovine-bovine cloning, yak-bovine, and takin-bovine cloning (48 vs. 28% vs. 5%, P < 0.01), and (4) cell numbers in the blastocysts of the three species of embryos were not different. These results suggest that the bovine oocytes can reprogram the takin, yak, and bovine fibroblast nuclei. However, the development efficiency of intra-species cloning tends to be higher than inter-species cloning; the more close the species of the donor cell is to the recipient oocyte (yak versus takin), the greater the blastocyst development in vitro. PMID- 16220544 TI - Tyrphostin-A47 inhibitable tyrosine phosphorylation of flagellar proteins is associated with distinct alteration of motility pattern in hamster spermatozoa. AB - To acquire fertilizing potential, mammalian spermatozoa must undergo capacitation and acrosome reaction. Our earlier work showed that pentoxifylline (0.45 mM), a sperm motility stimulant, induced an early onset of hamster sperm capacitation associated with tyrosine phosphorylation of 45-80 kDa proteins, localized to the mid-piece of the sperm tail. To assess the role of protein tyrosine phosphorylation in sperm capacitation, we used tyrphostin-A47 (TP-47), a specific protein tyrosine kinase inhibitor. The dose-dependent (0.1-0.5 mM) inhibition of tyrosine phosphorylation by TP-47 was associated with inhibition of hyperactivated motility and 0.5 mM TP-47-treated spermatozoa exhibited a distinct circular motility pattern. This was accompanied by hypo-tyrosine phosphorylation of 45-60 kDa proteins, localized to the principal piece of the intact-sperm and the outer dense fiber-like structures in detergent treated-sperm. Sperm kinematic analysis (by CASA) of spermatozoa, exhibiting circular motility (at 1st hr), showed lower values of straight line velocity, curvilinear velocity and average path velocity, compared to untreated controls. Other TP-47 analogues, tyrphostin AG1478 and -AG1296, had no effect either on kinematic parameters or sperm protein tyrosine phosphorylation. These studies indicate that TP-47-induced circular motility of spermatozoa is compound-specific and that the tyrosine phosphorylation status of 45-60 kDa flagellum-localized proteins could be key regulators of sperm flagellar bending pattern, associated with the hyperactivation of hamster spermatozoa. PMID- 16220545 TI - Survey of the year 2004: literature on applications of isothermal titration calorimetry. AB - The market for commercially available isothermal titration calorimeters continues to grow as new applications and methodologies are developed. Concomitantly the number of users (and abusers) increases dramatically, resulting in a steady increase in the number of publications in which isothermal titration calorimetry (ITC) plays a role. In the present review, we will focus on areas where ITC is making a significant contribution and will highlight some interesting applications of the technique. This overview of papers published in 2004 also discusses current issues of interest in the development of ITC as a tool of choice in the determination of the thermodynamics of molecular recognition and interaction. PMID- 16220546 TI - C/EBP homologous protein is necessary for normal osteoblastic function. AB - C/EBP homologous protein (CHOP) suppresses adipogenesis and accelerates osteoblastogenesis in vitro. However, the effects of CHOP in the skeleton in vivo are not known. To investigate the actions of CHOP on bone remodeling, we examined the skeletal phenotype of chop null mice from 1 to 12 months of age. Chop null mice appeared normal and their growth and serum insulin like growth factor (IGF) I and osteocalcin levels were normal. X-ray analysis of the skeleton revealed no abnormalities and bone mineral density was normal. Static and dynamic histomorphometry revealed that chop null mice had decreased bone formation rates, without changes in osteoblast cell number, indicating an osteoblastic functional defect. The number of osteoblasts and osteoclasts and eroded surface were normal. Northern blot analysis revealed decreased type I collagen and osteocalcin mRNA levels in calvariae of chop null mice. In conclusion, chop null mice exhibit decreased bone formation and impaired osteoblastic function, indicating that CHOP is necessary for the normal expression of the osteoblastic phenotype. PMID- 16220547 TI - Modifications of orientational dependence of microscopic magnetic resonance imaging T(2) anisotropy in compressed articular cartilage. AB - PURPOSE: To investigate the compression-induced changes in the orientational characteristics in T(2) anisotropy of articular cartilage using microscopic magnetic resonance imaging (microMRI). MATERIALS AND METHODS: Six beagle specimens were subjected to various levels of strain (0% to 27%) and were imaged at a minimum of two orientations (0 degrees and 55 degrees ). Two specimens at 14% and 27% strain were imaged at every 5 degrees increment over the first quadrant of the angular space. Quantitative two-dimensional T(2) images and three dimensional T(2) anisotropy maps of cartilage were constructed at a 19.8-microm in-depth resolution. RESULTS: The load-induced laminar appearance of cartilage at the magic angle became more distinct as the strain level increased. T(2) anisotropy maps of cartilage at 14% and 27% strain exhibited load-induced modifications in the collagen fibril ultrastructure, with a new peak toward the cartilage-bone interface and alterations to orientational dependence of T(2) anisotropy. CONCLUSION: Distinct alternations in the orientational dependence of microMRI T(2) anisotropy reflect the organizational modification of the collagen matrix due to external loading. This approach could become useful in detecting changes in cartilage's macromolecular structure due to injury or diseases. PMID- 16220548 TI - Urinary hepcidin in congenital chronic anemias. AB - BACKGROUND: Hepcidin, a regulator for iron homeostasis, is induced by inflammation and iron burden and suppressed by anemia and hypoxia. This study was conducted to determine the hepcidin levels in patients with congenital chronic anemias. PROCEDURE: Forty-nine subjects with anemia, varying degrees of erythropoiesis and iron burden were recruited. Eight children with immune thrombocytopenia were included as approximate age-matched controls. Routine hematologic labs and urinary hepcidin (uhepcidin) levels were assessed. For thalassemia major (TM) patients, uhepcidin was obtained pre- and post transfusion. RESULTS: In TM, uhepcidin levels increased significantly after transfusion, demonstrated wide variance, and the median did not significantly differ from controls or thalassemia intermedia (TI). In both thalassemia syndromes, the hepcidin to ferritin ratio, a marker of the appropriateness of hepcidin expression relative to the degree of iron burden, was low compared to controls. In TI and sickle cell anemia (SCA), median uhepcidin was low compared to controls, P = 0.013 and <0.001, respectively. In thalassemia subjects, uhepcidin levels were positively associated with ferritin. In subjects with SCA, uhepcidin demonstrated a negative correlation with reticulocyte count. CONCLUSIONS: This study examines hepcidin levels in congenital anemias. In SCA, hepcidin was suppressed and inversely associated with erythropoietic drive. In thalassemic syndromes, hepcidin was suppressed relative to the degree of iron burden. Transfusion led to increased uhepcidin. In thalassemia, the relative influence of known hepcidin modifiers was more difficult to assess. In thalassemic syndromes where iron overload and anemia have opposing effects, the increased erythropoietic drive may positively influence hepcidin production. PMID- 16220549 TI - St. Jude Children's Research Hospital, Memphis, Tennessee: gestational choriocarcinoma. AB - This paper describes a recent tumor board presentation conducted at our institution involving an adolescent with gestational choriocarcinoma. Despite its rarity in pediatrics, gestational choriocarcinoma offers unique diagnostic, treatment and off-therapy considerations in adolescent patients who have become pregnant. Key features and findings of the case as well as important management issues will be discussed. PMID- 16220550 TI - Inhibitory effect of minocycline on amyloid beta fibril formation and human microglial activation. AB - Minocycline, a derivative of the antibiotic tetracycline, displays neuroprotective properties in various models of neurodegenerative diseases and is now used in clinical trials, because of its relative safety and tolerability. Minocycline passes the blood-brain barrier and is presumed to inhibit microglial activation. In Alzheimer's disease brain, a number of proteins, including serum amyloid P component (SAP) and complement factors such as C1q, accumulate in amyloid beta (Abeta) plaques. In a previous study, SAP and C1q were found to be required for clustering of activated microglia in Abeta plaques. Furthermore, SAP and C1q enhanced Abeta fibril formation and Abeta mediated cytokine release by human microglia in vitro. In the present study, we report that tetracycline and minocycline dose-dependently reduce TNF-alpha and IL-6 release by adult human microglia upon stimulation with a combination of Abeta, SAP, and C1q. In addition, minocycline and to a lesser extent tetracycline inhibit fibril formation of Abeta as determined in a thioflavin-S-based fluorescence test. This inhibitory effect was observed with Abeta alone as well as with Abeta in combination with SAP and C1q. Our data suggest that minocycline and tetracycline at tolerable doses can inhibit human microglial activation. This activity in part is exerted by inhibition of (SAP and C1q enhanced) Abeta fibril formation. PMID- 16220551 TI - Rapid and accurate determination of MYCN copy number and 1p deletion in neuroblastoma by quantitative PCR. AB - MYCN amplification and 1p36 deletion are adverse prognostic factors in neuroblastoma, and rapid accurate determination of MYCN amplification is essential for risk stratification. MYCN copy number and 1p36 deletion status were determined by fluorescence in situ hybridization (FISH) and real time PCR in a diagnostic pathology laboratory setting on 35 consecutive patients with neuroblastoma. The PCR technique was technically successful in all cases and results were generally available within 24 hr of biopsy. There was no discordance between FISH and PCR results. Real time PCR is a reliable, accurate, and simple technique that can be applied to small neuroblastoma biopsies allowing rapid diagnosis. PMID- 16220552 TI - Primary pediatric brain tumors: statistics of Taipei VGH, Taiwan (1975-2004). AB - BACKGROUND: The purpose of the current study was to investigate a hospital series of 986 cases of primary pediatric brain tumors in Taiwan. METHODS: The authors reviewed the database of primary pediatric brain tumors in patients < 18 years of age collected in Taipei Veterans General Hospital (Taipei VGH) from 1975 to May 2004. Age and gender distribution, location, and classification of brain tumors were analyzed. Intracranial tumors with diagnostic imaging were included. Nontumoral lesions, cysts, and vascular malformations were excluded. RESULTS: The mean age of these 986 patients was 7.8 years, and the male to female ratio was 1.4:1. Supratentorial (including pineal-quadrigeminal) located tumors (58.3%) was predominant to infratentorial tumors (41.1%). In these series, 886 patients had either histologic diagnosis (842 patients) or clinical diagnosis (44 patients). The most common 5 categories of tumors were astrocytic tumors (31.1%), germ cell tumors (14.0%), medulloblastomas (13.3%), craniopharyngiomas (8.3%), and ependymal tumors (5.8%). Atypical teratoid/rhabdoid tumors (AT/RTs), a rare but highly malignant tumor, were 2.1%. The high incidence of primary intracranial germ cell tumors correlated with reported series from Japan and Korea. For the remaining 100 patients without diagnostic classifications, the majority were most likely astrocytic tumors in brain stem. CONCLUSIONS: The authors analyzed a large hospital series of primary brain tumors in children. Both histologically verified and unverified tumors were recruited to avoid selective bias. Although it was not a study of a population-based brain tumor registry, it could still be representative of primary pediatric brain tumors in Taiwan. PMID- 16220554 TI - Negative thoughts after childbirth: development and preliminary validation of a self-report scale. AB - This study describes the development and initial validation of a questionnaire that is suitable for detecting and measuring postpartum negative thoughts. Semistructured interviews with mothers who had suffered from postnatal depression were conducted to inform the content of the questionnaire. The initial questionnaire, alongside other measures, was then administered to a nonclinical sample of mothers with babies aged 0-7 months. Using principal components analysis, a two-factor structure was obtained for the Postnatal Negative Thoughts Questionnaire (PNTQ). The factors included appraisal of cognition, emotion, and situation (ACES) and baby-related and motherhood negative thoughts (BRM-NT). The psychometric properties demonstrated acceptable validity, satisfactory test retest reliability, and internal consistency. These findings suggest that the PNTQ is a reliable and valid measure for assessing postpartum negative thoughts. Consistent with previous research, findings also suggest that appraisal of negative thoughts is more strongly related to postpartum depression than to the experience of negative thoughts per se. Clinicians may use the PNTQ to offer new mothers the opportunity to assess whether negative thoughts or metacognitive appraisals are being experienced as problematic. Additionally, a direct focus upon the metacognitive appraisals of postpartum negative thoughts may provide a useful adjunct to traditional cognitive therapy approaches. Recommendations for future research are discussed. PMID- 16220553 TI - Mutation rates at Y chromosome specific microsatellites. AB - A collaborative work was carried out by the Spanish and Portuguese ISFG Working Group (GEP-ISFG) to estimate Y-STR mutation rates. Seventeen Y chromosome STR loci (DYS19, DYS385, DYS389I and II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS460, DYS461, DYS635 [GATA C4], GATA H4, and GATA A10) were analyzed in a sample of 3,026 father/son pairs. Among 27,029 allele transfers, 54 mutations were observed, with an overall mutation rate across the 17 loci of 1.998 x 10(-3) (95% CI, 1.501 x 10(-3) to 2.606 x 10(-3)). With just one exception, all of the mutations were single-step, and they were observed only once per gametogenesis. Repeat gains were more frequent than losses, longer alleles were found to be more mutable, and the mutation rate seemed to increase with the father's age. Hum Mutat 26(6), 520-528, 2005. (c) 2005 Wiley-Liss, Inc. PMID- 16220555 TI - Prognostic factors for survival in patients with advanced renal cell carcinoma undergoing nonmyeloablative allogeneic stem cell transplantation. AB - BACKGROUND: The objective of this study was to identify prognostic factors for predicting survival in patients with advanced renal cell carcinoma (RCC) who had undergone an allogeneic stem cell transplantation after failure on immunotherapy. METHODS: The authors studied 70 patients with advanced RCC who underwent allogeneic transplantation with a fludarabine-based, reduced-intensity regimen. Ten parameters were analyzed at the time of transplantation for their power to predict survival. Clinical features were examined first univariately; then, variables that were correlated significantly with survival in the univariate analysis were included in a multivariate Cox regression model. RESULTS: Factors that were found to be associated significantly with limited survival were performance status, the number of metastatic sites, the presence of mediastinal metastasis, hemoglobin level, C-reactive protein (CRP) level, lactate dehydrogenase (LDH) level, and neutrophil counts. All these variables were included in a multivariate Cox regression model, and three were retained in the final model. Patients were classified according to the score estimated by the final Cox model in two groups (above or below the median value): The median survival was 3.5 months for patients who had a poor prognosis patients versus 23 months for patients who had a good prognosis. CONCLUSIONS: The current findings suggested that three easily available parameters (performance status, CRP level, and LDH level) could be used to stratify patients with advanced RCC who are candidates for allografting and to assist clinicians in decision-making and selection of an appropriate treatment program. PMID- 16220556 TI - Stereotactic radiosurgery (SRS): treatment option for recurrent glioblastoma multiforme (GBM). AB - BACKGROUND: This article describes the results of a study of stereotactic radiosurgery (SRS) in the treatment of patients with recurrent malignant glioma. METHODS: Thirty-two patients with recurrent glioblastoma multiforme (GBM) were treated for 36 lesions with SRS from 1993 to 2001. Nineteen patients were male and 13 were female. The median age at primary diagnosis of the tumor was 56 years (range, 33-76 yrs). At the time of initial diagnosis a total neurosurgical resection was performed in 7, a subtotal resection in 21, and a biopsy in 4 patients. Histology evaluations revealed glioblastoma multiforme (WHO Grade IV) in all 32 patients. In all patients radiotherapy was performed as the first-line therapy, applied as fractionated external beam radiotherapy. The median interval between primary irradiation and reirradiation was 10 months. The median dose applied was 15 Gy (range, 10-20 Gy) prescribed to the 80% isodose line that encompassed the target volume. No concomitant chemotherapy was applied. RESULTS: Treatment was well tolerated by all patients. No acute toxicities > CTC Grade II occurred. No severe long-term toxicities including radionecrosis were observed. The median follow-up time was 13 months (range, 1-89 mo). All patients died of tumor progression during follow-up. The median overall survival from primary diagnosis of the tumor was 22 months (range, 9-133 mo). The survival rate at 1 year was 90%, and 49% and 26% at 2 and 3 years, respectively. Median overall survival after SRS was 10 months. At 6 and 12 months after SRS, survival rates were 72% and 28%, respectively. Median progression-free survival after SRS was 7 months. CONCLUSIONS: SRS offers effective treatment as a salvage therapy for a subgroup of patients with smaller lesions of recurrent GBM. PMID- 16220557 TI - Identification of 29 novel and nine recurrent fibrillin-1 (FBN1) mutations and genotype-phenotype correlations in 76 patients with Marfan syndrome. AB - Marfan syndrome (MFS) is an autosomal-dominant disorder of the fibrous connective tissue that is typically caused by mutations in the gene coding for fibrillin-1 (FBN1), a major component of extracellular microfibrils. The clinical spectrum of MFS is highly variable and includes involvement of the cardiovascular, skeletal, ocular, and other organ systems; however, the genotype-phenotype correlations have not been well developed. Various screening methods have led to the identification of about 600 different mutations (FBN1-UMD database; www.umd.be). In this study we performed SSCP and/or direct sequencing to analyze all 65 exons of the FBN1 gene in 116 patients presenting with classic MFS or related phenotypes. Twenty-nine novel and nine recurrent mutations were identified in 38 of the analyzed patients. The mutations comprised 18 missense (47%), eight nonsense (21%), and five splice site (13%) mutations. Seven further mutations (18%) resulted from deletion, insertion, or duplication events, six of which led to a frameshift and subsequent premature termination. Additionally, we describe new polymorphisms and sequence variants. On the basis of the data presented here and in a previous study, we were able to establish highly significant correlations between the FBN1 mutation type and the MFS phenotype in a group of 76 mutation-positive patients for whom comprehensive clinical data were available. Most strikingly, there was a significantly lower incidence of ectopia lentis in patients who carried a mutation that led to a premature termination codon (PTC) or a missense mutation without cysteine involvement in FBN1, as compared to patients whose mutations involved a cysteine substitution or splice site alteration. PMID- 16220558 TI - Sexual functioning in patients with hepatocellular carcinoma. AB - BACKGROUND: The objectives of the current study were to assess the rates of sexual dysfunction in patients with hepatocellular carcinoma (HCC) and compare the rates of sexual morbidity with a sample of patients diagnosed with chronic liver disease (CLD) and the general population. It was expected that patients diagnosed with HCC would have a greater prevalence of sexual dysfunction than those diagnosed with CLD and the general population, respectively. Furthermore, those patients who reported a sexual dysfunction would also have a poorer quality of life (QOL). METHODS: Twenty-one men diagnosed with HCC and 23 men diagnosed with CLD completed a battery of questionnaires that included the Sexual History Questionnaire and the Functional Assessment of Cancer Therapy-Hepatobiliary. RESULTS: Results indicated that 41% of patients reported a current sexual problem (29% of HCC patients and 71% of patients with CLD) and 43% met the Diagnostic and Statistical Manual of Mental Disorders-4th edition (DSM-IV) criteria for at least 1 type of sexual dysfunction (25% of patients with HCC and 75% of patients with LD). Of the total sample, 68% reported being diagnosed with a comorbid medical condition or taking a medication that had potential sexual side effects. After eliminating cases with comorbid medical conditions and/or who were taking medications that contributed to sexual morbidity, the rate of sexual problems were found to be similar to that of the general population. Clinically significant differences were found with regard to the QOL. People who were experiencing sexual problems also reported a poorer QOL. CONCLUSIONS: Although a large percentage of sexual dysfunction may have been secondary to comorbid medical problems and medications, treatment of the sexual dysfunction is still warranted. PMID- 16220559 TI - Matrix metalloproteinases in carcinoma of unknown primary. AB - BACKGROUND: The purpose was to study proteolysis-related molecules, matrix metalloproteinase-2 (MMP-2) and MMP-9 and tissue inhibitor of metalloproteinases 1 (TIMP-1), in carcinoma of unknown primary (CUP). METHODS: Paraffin-embedded tumor material from 75 patients diagnosed with CUP was used. Tumor histologies were adenocarcinoma (77%), undifferentiated carcinoma (19%), and squamous cell carcinoma (4%) and patients were categorized into favorable (62%) and unfavorable (38%) subsets. The tissue expression of MMP-2, MMP-9, and TIMP-1 was assessed by use of specific monoclonal antibodies and evaluated by means of a visual staining score. The expression of molecules studied was analyzed against clinicopathological data. RESULTS: MMP-2 was found expressed in 69% (strong expression in 49%), MMP-9 in 49% (strong in 36%), and TIMP-1 in 79% (strong in 44%) of studied cases. The expression of MMP-2 correlated positively with MMP-9. TIMP-1 was significantly higher in unfavorable compared with favorable tumors and was associated with a shorter survival of patients (7.5 vs. 12 mos). No other associations were detected. CONCLUSIONS: MMP-2, MMP-9, and TIMP-1 are widely expressed in CUP, suggesting an essential role of proteolysis in these tumors. TIMP-1 may be considered a possible marker of poor prognosis in CUP patients. PMID- 16220560 TI - On the importance of carbohydrate-aromatic interactions for the molecular recognition of oligosaccharides by proteins: NMR studies of the structure and binding affinity of AcAMP2-like peptides with non-natural naphthyl and fluoroaromatic residues. AB - The specific interaction of a variety of modified hevein domains to chitooligosaccharides has been studied by NMR spectroscopy in order to assess the importance of aromatic-carbohydrate interactions for the molecular recognition of neutral sugars. These mutant AcAMP2-like peptides, which have 4-fluoro phenylalanine, tryptophan, or 2-naphthylalanine at the key interacting positions, have been prepared by solid-phase synthesis. Their three-dimensional structures, when bound to the chitin-derived trisaccharide, have been deduced by NMR spectroscopy. By using DYANA and restrained molecular dynamics simulations with the AMBER 5.0 force field, the three-dimensional structures of the protein-sugar complexes have been obtained. The thermodynamic analysis of the interactions that occur upon complex formation have also been carried out. Regarding binding affinity, the obtained data have permitted the deduction that the larger the aromatic group, the higher the association constant and the binding enthalpy. In all cases, entropy opposes binding. In contrast, deactivation of the aromatic rings by attaching fluorine atoms decreases the binding affinity, with a concomitant decrease in enthalpy. The role of the chemical nature of the aromatic ring for establishing sugar contacts has been thus evaluated. PMID- 16220561 TI - Electron-rich diferrous-phosphane-thiolates relevant to Fe-only hydrogenase: is cyanide "nature's trimethylphosphane"? AB - The two-step one-pot oxidative decarbonylation of [Fe2(S2C2H4)(CO)4(PMe3)2] (1) with [FeCp2]PF6, followed by addition of phosphane ligands, led to a series of diferrous dithiolato carbonyls 2-6, containing three or four phosphane ligands. In situ measurements indicate efficient formation of 1(2+) as the initial intermediate of the oxidation of 1, even when a deficiency of the oxidant was employed. Subsequent addition of PR3 gave rise to [Fe2(S2C2H4)(mu CO)(CO)3(PMe3)3]2+ (2) and [Fe2(S2C2H4)(mu-CO)(CO)2(PMe3)2(PR3)2]2+ (R = Me 3, OMe 4) as principal products. One terminal CO ligand in these complexes was readily substituted by MeCN, and [Fe2(S2C2H4)(mu-CO)(CO)2(PMe3)3(MeCN)]2+ (5) and [Fe2(S2C2H4)(mu-CO)(CO)(PMe3)4(MeCN)]2+ (6) were fully characterized. Relevant to the H(red) state of the active site of Fe-only hydrogenases, the unsymmetrical derivatives 5 and 6 feature a semibridging CO ligand trans to a labile coordination site. PMID- 16220562 TI - Mechanistic investigation of the 2,5-diphenylpyrrolidine-catalyzed enantioselective alpha-chlorination of aldehydes. AB - The mechanism for the 2,5-diphenylpyrrolidine-catalyzed enantioselective alpha chlorination of aldehydes with electrophilic halogenation reagents has been investigated by using experimental and computational methods. These studies have led us to propose a mechanism for the reaction that proceeds through an initial N chlorination of the chiral catalyst-substrate complex, followed by a 1,3 sigmatropic shift of the chlorine atom to the enamine carbon atom. The suggested reaction course is different from previously proposed mechanisms for organocatalytic enamine reactions, in which the carbon-electrophile bond is formed directly. Furthermore, the rate-determining step in the overall reaction was determined and the presence of nonlinear effects was probed. PMID- 16220563 TI - Metal aminocarboxylate coordination polymers with chain and layered structures. AB - The synthesis and structures of metal aminocarboxylates prepared in acidic, neutral, or alkaline media have been explored with the purpose of isolating coordination polymers with linear chain and two-dimensional layered structures. Metal glycinates of the formulae [CoCl2(H2O)2(CO2CH2NH3)] (I), [MnCl2(CO2CH2NH3)2] (II), and [Cd3Cl6(CO2CH2NH3)4] (III) with one-dimensional chain structures have been obtained by the reaction of the metal salts with glycine in an acidic medium under hydro/solvothermal conditions. These chain compounds contain glycine in the zwitterionic form. 4-Aminobutyric acid transforms to a cyclic amide under such reaction conditions, and the amide forms a chain compound of the formula [CdBr2(C4H7NO)2] (IV). Glycine in the zwitterionic form also forms a two-dimensional layered compound of the formula [Mn(H2O)2(CO2CH2NH3)2]Br2 (V). 6-Aminocaproic acid under alkaline conditions forms layered compounds with metals at room temperature, the metal being coordinated both by the amino nitrogen and the carboxyl oxygen atoms. Of the two layered compounds [Cd{CO2(CH2)5NH2}2]2 H2O (VI) and [Cu{CO2(CH2)5NH2}2]2 H2O (VII), the latter has voids in which water molecules reside. PMID- 16220564 TI - Bioactive compounds from Rhodiola rosea (Crassulaceae). AB - The methanol extract of the underground part of Rhodiola rosea was found to show inhibitory activity against Staphylococcus aureus. Bioactivity-guided fractionation of a 95% ethanol extract from the stems of R. rosea led to the isolation of five compounds: gossypetin-7-O-L-rhamnopyranoside (1), rhodioflavonoside (2), gallic acid (3), trans-p-hydroxycinnamic acid (4) and p tyrosol (5). Their structures were elucidated by UV, IR, MS and NMR data, as well as by comparison with those of the literature. Compounds 1 and 2 were evaluated for their antibacterial and antiprostate cancer cell activities. Compounds 1 and 2 exhibited activity against Staphylococcus aureus with minimum inhibitory concentrations of 50 microg/mL and 100 microg/mL, respectively. Cytotoxicity studies of 1 and 2 also displayed activity against the prostate cancer cell line with IC(50) values of 50 microg/mL and 80 microg/mL, respectively. PMID- 16220565 TI - The hepatoprotective action of ten herbal extracts in CCl4 intoxicated liver. AB - The effect of ten phytotherapeutic products on CCl(4) intoxicated liver in albino male Wistar rats was investigated. Biochemical parameters, including serum transaminase activity (GPT and GOT), histoenzymological measurements (lactate dehydrogenase, LDH; succinate dehydrogenase, SDH, cytochromoxidase, CyOx; Mg(2+) dependent adenosine triphosphatase, ATP-ase) and histochemical (Sudan black) and histological examinations (haematoxylin-eosin staining) of the liver were investigated. Some positive effects such as the reduction of hepatocytolysis and steatosis, and a return to normal values of the activity of some enzymes in the following plants: Chrysanthemum balsamita, Echinacea pallida, Calendula officinalis and Corylus avelana were obtained. PMID- 16220566 TI - Gastrointestinal stimulatory and uterotonic activities of dietary radish leaves extract are mediated through multiple pathways. AB - Raphanus sativus, commonly known as radish, is a food plant known worldwide for its culinary and medicinal properties especially as a laxative and abortifacient. This study reports the gastrointestinal and uterine tone modulatory activities of the crude extract (Rl.Cr) of radish leaves. Rl.Cr, showing the presence of saponins and alkaloids, exhibited a spasmogenic effect (0.03-10 mg/mL) in isolated rabbit jejunum, rat stomach fundus and uterus which was partially blocked by atropine. In contrast, Rl.Cr was found to be devoid of any stimulatory effect in rat ileum, instead showed an inhibitory effect (0.1 mg/mL) on the ACh dose-response curves. A mild relaxant effect was also observed in rabbit jejunum at the lower doses (0.1-0.3 mg/mL) but not against K(+)-induced contractions, ruling out a calcium channel blocking effect. In guinea-pig ileum, Rl.Cr exhibited a stimulant effect resistant to atropine while sensitive to pyrilamine pretreatment. The aqueous fraction, showing a strong presence of saponins, was found to be more efficacious than the non-polar fractions in its spasmogenic effect. This study shows the presence of species-dependent gastrointestinal effects of radish mediated partially through cholinergic receptors in rabbit and rat tissues, but through histaminergic activation in the guinea-pig, providing a scientific basis for its use in gut and uterine affections while also giving a wider picture of the activity profile of radish by using different species of animals. PMID- 16220567 TI - Ultrastructural changes in Leydig cells and cauda epididymal spermatozoa induced by Azadirachta indica leaves in albino rats. AB - The effects of Azadirachta indica leaves (500 mg/kg body weight, orally/day), testosterone (0.25 mg/kg body weight/day, intramuscularly) and a combination of these two were investigated as to whether Azadirachta indica leaves affect the morphology of Leydig cells and epididymal sperm and their organelles at the ultrastructural level. Azadirachta indica treated rats showed atrophic seminiferous tubules with widening intercellular spaces. Leydig cells exhibited characteristics of degeneration such as indented nuclei; the commencement of degeneration was evident from less chromatin; the reduced amount of a marked decrease in organelle content and scarcity of other cell types of the interstitium in highly vacuolated cytoplasm, which were seem from both light and electron microscopic observations. The number of Leydig cells and their nuclear diameter were reduced significantly. Pathological changes in the spermatozoa of the cauda epididymis were observed and the spermatozoa retained cytoplasmic droplets. It is suggested that regression of Leydig cells and the absence of interstitium cell types indicates an androgen deficiency, which affects the spermatozoa in the epididymis by disturbing the internal epididymal milieu. The antiandrogenic and antispermatogenic properties of Azadirachta indica leaves appear to lead to a reduced fertilizing ability of the sperm. PMID- 16220568 TI - The anti-cancer effects of poi (Colocasia esculenta) on colonic adenocarcinoma cells In vitro. AB - Hawaiians tend to have lower incidence rates of colorectal cancer and it was hypothesized that this may be due to ethnic differences in diet, specifically, their consumption of poi, a starchy paste made from the taro (Colocasia esulenta L.) plant corm. Soluble extracts of poi were incubated at 100 mg/mL in vitro for antiproliferative activity against the rat YYT colon cancer cell line. (3)H thymidine incorporation studies were conducted to demonstrate that the poi inhibited the proliferation of these cancer cells in a dose-dependent manner. The greatest suppression of YYT colon cancer growth occurred when 25% concentration was used. When poi was incubated with the YYT cells after 2 days, the YYT cells underwent apoptotic changes as evidenced by a positive terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) stain. Poi enhanced the proliferation of normal mouse splenocyte control cells, suggesting that poi is not simply toxic to all cells but even has a positive immunostimulatory role. By flow cytometry, T cells (CD4+ and CD8+) were predominantly activated by the poi. Although numerous factors can contribute to the risk of colon cancer, perhaps poi consumption may contribute to the lower colon cancer rates among Hawaiians by two distinct mechanisms. First, by inducing apoptosis within colon cancer cells; second, by non-specifically activating lymphocytes, which in turn can lyse cancerous cells. Our results suggest for the first time that poi may have novel tumor specific anti-cancer activities and future research is suggested with animal studies and human clinical trials. PMID- 16220570 TI - Antibacterial and antioxidant activity of Sutherlandia frutescens (Fabaceae), a reputed anti-HIV/AIDS phytomedicine. AB - Dried ground leaves of Sutherlandia frutescens were extracted by both sequential extraction with four solvents, starting with the least polar and separately with acetone, ethanol and water. The extracts were tested for antibacterial and antioxidant activity. The hexane extract was, generally, the most active extract against S. aureus, E. faecalis and E. coli with MIC values of 0.31, 1.25 and 2.50 mg/mL, respectively. The second method extracted compounds with antioxidant activity as shown by the DPPH free-radical scavenging assay. The use of Sutherlandia frutescens for topical staphylococcal infections, when formulated in an oily base appears to have a rational basis. PMID- 16220569 TI - Antarth, a polyherbal preparation protects against the doxorubicin-induced toxicity without compromising its Antineoplastic activity. AB - Doxorubicin (DOX), an anthracycline drug widely used for the treatment of various cancers, causes a cumulative dose-dependent cardiotoxicity that is characterized by an irreversible dilated cardiomyopathy and congestive heart failure. Antarth (ANT) a polyherbal preparation was evaluated for its cardioprotective properties against doxorubicin-induced cardiotoxicity in mice. Mice were treated with 25 mg/kg ANT orally once daily for 5 consecutive days before a single intraperitoneal injection of 15 mg/kg doxorubicin. The animals were killed 30 h after DOX treatment. DOX induced a significant elevation in the serum levels of glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), creatine kinase (CK-MB) and lactate dehydrogenase (LDH), indicating its acute cardiotoxicity. The treatment of mice with ANT before DOX administration significantly reduced the serum levels of GPT, GOT, CK-MB and LDH indicating that ANT protected against the DOX-induced cardiotoxicity. Pretreatment of mice with 25 mg/kg ANT inhibited the DOX-induced decline in the antioxidant status. Intraperitoneal injection of 1.25 mg/kg DOX once daily for 9 consecutive days significantly improved the survival of mice bearing Ehrlich ascites carcinoma (EAC). Treatment of EAC with 25 mg/kg ANT alone did not affect the anticancer activity of DOX since ANT did not alter the tumor cell growth, the median survival time and average survival time of tumor bearing mice. The present study demonstrates that ANT protects mice against DOX-induced cardiotoxicity, without compromising the antineoplastic activity of DOX. PMID- 16220571 TI - Structure-activity relationship for antiinflammatory effect of luteolin and its derived glycosides. AB - Luteolin and its derived glycosides such as a cynaroside, cesioside, isoorientin and stereolensin have been isolated and identified from different kinds of plant species. A (13)C NMR spectroscopic analysis of stereolensin has been done for the first time. These structurally related flavonoids were examined in vitro for their abilities to inhibit enzymes for the synthesis of thromboxane B(2) and leukotriene B(4) as well as hydrogen peroxide scavenging activity. Luteolin exhibited a high inhibitory activity against both thromboxane and leukotriene synthesis. In particular, a remarkable inhibitory effect was observed against leukotriene enzyme activity. The glycosides, cynaroside and cesioside, possessed a moderate inhibition activity against both enzyme synthesis pathways, while isoorientin and stereolensin exhibited selectively good activity against thromboxane synthesis. All the flavonoids showed excellent scavenging activity for the hydrogen peroxide at all the concentrations tested. The results demonstrated that the reactivities of luteolin and its related glycosides against arachidonic acid synthesis and hydrogen peroxide scavenging are dependent on their molecular structures. The presence of ortho-dihydroxy groups at the B ring and OH substitution pattern at C-5 position of the A ring could significantly contribute to the antiinflammatory and antioxidant activities of flavonoids. PMID- 16220573 TI - Concurrent administration of aqueous Azadirachta indica (neem) leaf extract with DOCA-salt prevents the development of hypertension and accompanying electrocardiogram changes in the rat. AB - The effect of concurrent administration of Azadirachta indica leaf extract with DOCA-salt was investigated in the development of hypertension.Over 5-6 week old, inbred male Wistar rats with a starting weight of 190 g were given either: (1) twice weekly subcutaneous (s.c.) injections of vehicle (soyabean oil, 0.25 mL per animal) for the first 2 weeks, plus normal drinking water (controls); (2) twice weekly (s.c.) injections (weeks 1 and 2 only) of 15 mg/kg DOCA dissolved in vehicle, plus drinking water containing 1.0% NaCl and 0.03% KCl (DOCA-salt group); or (3) 20 mg/kg of aqueous neem extract daily, in addition to the DOCA salt treatment (DOCA-salt-neem group). All groups (8-12 animals) received normal rat pellets ad libitum and their BP was measured weekly. Terminally, the animals were anaesthetized and ECGs recorded using s.c. pins in a lead II configuration. The mean arterial pressure was significantly lower (p < 0.05) in the control (97 +/- 3.7 mmHg) and DOCA-salt-neem (87 +/- 3.4 mm Hg) groups than in the DOCA-salt group (115 +/- 7.1 mm Hg). PR and RR intervals and the duration of the QRS complex were shorter (p < 0.05) in the DOCA-salt group than in the control and DOCA-salt-neem groups. Amplitude of the QRS complex was increased (p < 0.05) in the DOCA-salt group compared with both the DOCA-salt-neem and the control groups.Daily administration of 20 mg/kg neem-leaf extract concurrently with DOCA salt for 5 weeks, prevents the development of hypertension and the accompanying alterations in the ECG patterns seen in DOCA-salt treated rats. PMID- 16220572 TI - Chemical and antibacterial evaluation of Hypericum triquetrifolium Turra. AB - Five extracts and pure compounds from the aerial parts of Hypericum triquetrifolium were tested for their antibacterial activity against 31 gram positive and gram-negative strains using the agar dilution method (Lorian). The ethyl acetate extract exhibited a weak antibacterial activity against Staphylococcus strains, and pure constituents such as quercetin and I3,II8 biapigenin were the active components of this extract. PMID- 16220574 TI - Opuntia ficus indica (L.) Mill. fruit juice protects liver from carbon tetrachloride-induced injury. AB - The protective effects of the juice of Opuntia ficus indica fruit (prickly pear) against carbon tetrachloride (CCl(4))-induced hepatotoxicity were examined in rats. The animals were treated orally with the juice (3 mL/rat) 2 h after administration of the hepatotoxic agent. Preventive effects were studied by giving the juice (3 mL/rat) for 9 consecutive days. On day 9 the rats received the hepatotoxic agent. Morphological and biochemical evaluations were carried out 24, 48 and 72 h after induction of the hepatic damage. Data show that O. ficus indica fruit juice administration exerts protective and curative effects against the CCl(4)-induced degenerative process in rat liver. Histology evaluation revealed a normal hepatic parenchyma at 48 h; the injury was fully restored after 72 h. Moreover, a significant reduction in CCl(4)-induced increase of GOT and GPT plasma levels is evident; these data are in agreement with the functional improvement of hepatocytes. O. ficus indica fruit juice contains many phenol compounds, ascorbic acid, betalains, betacyanins, and a flavonoid fraction, which consists mainly of rutin and isorhamnetin derivatives. Hepatoprotection may be related to the flavonoid fraction of the juice, but other compounds, such as vitamin C and betalains could, synergistically, counteract many degenerative processes by means of their antioxidant activity. PMID- 16220575 TI - Pharmacokinetics of valerenic acid after administration of valerian in healthy subjects. AB - OBJECTIVES: To describe the pharmacokinetics of valerenic acid in a group of healthy adults after a single oral dose of valerian using a newly developed sensitive assay for serum concentrations of valerenic acid, a commonly used marker for qualitative and quantitative analysis of valerian root and valerian products. STUDY DESIGN: Six healthy adults (22-61 years, five men, one female) received a single 600 mg dose of valerian at 08:00. Blood samples were collected for 8 h after administration. Valerenic acid was extracted from serum and measured using a LC/MS/MS method developed in our laboratory. RESULTS: The maximum serum concentration of valerenic acid for five of the six subjects occurred between 1 and 2 h ranging from 0.9 to 2.3 ng/mL. Valerenic acid serum concentrations were measurable for at least 5 h after the valerian dose. One subject showed a peak plasma value at 1 h and a second peak at 5 h. The elimination half-life (T(1/2)) for valerenic acid was 1.1 +/- 0.6 h. The area under the concentration time curve (AUC) as a measure of valerenic acid exposure was variable (4.80 +/- 2.96 microg/mL. h) and not correlated with subject's age or weight. CONCLUSIONS: Assuming that valerenic acid serum concentrations correlate with the pharmacological activity of valerian, the timing of the valerenic acid peak concentration is consistent with the standard dosage recommendation to take valerian 30 min to 2 h before bedtime. Ongoing studies are evaluating the relationship between valerenic acid serum concentrations and objective measures of sleep in patients. PMID- 16220576 TI - Activity of aqueous extract of the bark of Vitex doniana on uterine muscle response to drugs. AB - The activity of Vitex doniana on the uterine muscle response was investigated. The bark of Vitex doniana was extracted in boiled water at 100 degrees C, and the extracted solution filtered and centrifuged with refrigeration. The extract prepared from the dry powder extract was tested on uterine muscle strip preparations. The bark extract of Vitex doniana was analysed elementally and found to contain much more potassium and phosphate than calcium, magnesium, zinc and iron. The presence of potassium ions in excess may also be partly responsible for the effect of the crude extract on uterine muscle activity. In another study, Vitex doniana extract induced graded uterine muscle contractions and also potentiated the contractile effects of prostaglandins, ergometrine and oxytocin. However, the potentiating effect was not significant on the contractile responses to acetylcholine and potassium chloride. The investigation therefore suggests that the effect of the Vitex doniana bark extract may be not only voltage operated but may act via uterotonic receptors. Therefore, the use of Vitex doniana to control postpartum bleeding after child birth may be justified. PMID- 16220577 TI - Antiinflammatory flavonoids from Opuntia dillenii (Ker-Gawl) Haw. flowers growing in Egypt. AB - Opuntia dillenii (Ker-Gawl) Haw. (Family Cactaceae), is used in folk medicine as an antidiabetic and antiinflammatory. The antiinflammatory activity of the alcohol extracts of the flowers, fruits and stems was carried out using the carrageenan-induced rat paw oedema model. The analgesic effect of the same extracts was evaluated using electric current as a noxious stimulus. The alcohol extract of the flowers revealed the most potent antiinflammatory effect and a pronounced analgesic action at a dose of 200 mg/kg. Bioassay-guided fractionation of this extract using VLC followed by Sephadex and paper chromatography, afforded three flavonoid glycosides, namely, kaempferol 3-O-alpha-arabinoside, isorhamnetin-3-O-glucoside and isorhamnetin-3-O-rutinoside. Their identification was based on physical, chemical and spectroscopic data. PMID- 16220578 TI - Effects of nutritional supplement with herbal extract on bone mineral density and height in prepubescent children - a preliminary study. AB - The aim of this study was to investigate the efficacy of a nutritional supplement with herbal extracts on height and on the bone mineral density (BMD) development of prepubescent children who were in the 25th percentile of standard height of their age. All children were administered a supplement for 6 months, with height and BMD measured. The supplement increased significantly the height and BMD, compared with the baseline in both boys and girls. The annual growth rate was higher than the standard rate. These preliminary data indicate that supplements enhanced the rate of BMD and height development, although this result must be replicated in a large population-based study and placebo-controlled trials to confirm the conclusions. PMID- 16220579 TI - Toxic effects of chromatographic fractions of Phyllanthus amarus on the serum biochemistry of rats. AB - Chromatographic fractions obtained from Phyllanthus amarus were tested for toxicity on the serum biochemistry of rats. The results revealed that some fractions of P. amarus had potentially deleterious effects on the blood and therefore caution should be exercised in the use of P. amarus as a medicinal plant. PMID- 16220580 TI - Cytotoxic activities of ethyl acetate extract and a metabolite from a Monocillium species. AB - The ethyl acetate soluble fraction of a cultural broth of a Monocillium species afforded the isolation of 5-hydroxymethylfurfural. Both the extract and 5 hydroxymethylfurfural showed significant cytotoxic activities in a brine shrimp bioassay and the LC(50) values were found to be 14.96 microg/mL and 23.71 microg/mL, respectively. PMID- 16220581 TI - Chinese officials accused of forcing abortions in Shandong. PMID- 16220582 TI - Money for health in Russia, at long last. PMID- 16220583 TI - Rebuilding New Orleans. PMID- 16220584 TI - Lower urinary tract symptoms after total and subtotal hysterectomy: results of a randomized controlled trial. AB - The aim of this Danish multicenter trial was to compare the proportion of women with lower urinary tract symptoms after total abdominal hysterectomy (TAH) and subtotal abdominal hysterectomy (SAH) for benign uterine disorders. A total of 319 women were randomized to TAH (n = 158) or SAH (n = 161). Women were followed up for 1 year by strict data collection procedures, including postal questionnaires. Results were analyzed by intention-to-treat analyses. Urinary incontinence was found less often among TAH women than among SAH women. This was due to a larger reduction of the number of women with stress and urinary incontinence in the TAH group. No other differences were found between the two operation methods. The number of women with urinary incontinence and frequency was reduced from study entry for follow-up, while double/triple voiding was increased. Incontinent women had significantly lower quality of life scores than continent women PMID- 16220585 TI - Effects of different soil types on the Collembolans Folsomia candida and Hypogastrura assimilis using the herbicide Phenmedipham. AB - Soil ecotoxicology studies are usually performed in standard soils such as Organization for Economic Cooperation and Development artificial soil or LUFA ST. 2.2, a natural soil. When assessing the toxic effects in the environment, soil properties are often different from those in standard soils, which might lead to different exposure situation for the test species and therefore to misleading conclusions. Selected to cover a broad range of properties and based on the Euro Soils concept, 17 different soils were studied regarding their suitability to two tes species: Folsomia candida and Hypogastrura assimilis (Collembola). In reproduction tests, the test species reacted differently to the soils. F. candida was less affected by soil properties: 500 to 1200 juveniles/vessel were found in untreated soils (i.e. controls). These differences can be attributed to normal interindividual variability. H. assimilis showed a significant correlation with maximum water-holding capacity and also a tendency to lower the reproductive output in soils with a low pH (<4). Therefore, some soils were revealed to be inappropriate for tests with H. assimilis. In the main tests, the effect of the reference test substance Phenmedipham (formulation Betosyp) was studied in those soils where sufficient reproduction was determined beforehand. Clearly, the chronic end point was more sensitive than survival when testing Phenmedipham. In H. assimilis, because of high variability and low effects of tested dosages, no conclusions could be drawn. In F. candida, different soils caused different toxic effects: Juveniles preferred soils with high C-to-N ratios. Higher microbial activity might support a quicker metabolization of the test substance. In general, the toxic response is caused by a synergistic action of several soil properties with each of them exerting an effect too small to be clarified with the available set of data. PMID- 16220586 TI - The role of radiotracer imaging in Parkinson disease. PMID- 16220587 TI - Higher resolution MRI and image modeling for predicting surgical outcome in partial epilepsy. PMID- 16220589 TI - Healthcare premiums doomed to outpace wages, inflation. PMID- 16220588 TI - [Course leader education of pediatric nurses and pediatric aides: autogenic training, progressive muscle relaxation and the five "Tibet" exercises for children]. PMID- 16220590 TI - Avoiding eye complications. PMID- 16220591 TI - Diabetes basics. Understanding insulin. PMID- 16220592 TI - Get to know grains. PMID- 16220593 TI - Staying independent with canine help. PMID- 16220594 TI - Supermarket smarts. Cheese. PMID- 16220595 TI - How hunger happens. The brain's role in obesity. PMID- 16220596 TI - Diabetes and your marriage. Making things work. PMID- 16220597 TI - Blood glucose monitoring. What do the numbers tell you? PMID- 16220598 TI - For parents. To pump, or not to pump? PMID- 16220599 TI - 7 tips for commuting with diabetes. PMID- 16220600 TI - Diabetes quiz. How much do you know about heart disease risk? PMID- 16220601 TI - Medicare's new drug program creates challenges. Part D is part drug coverage, part difficult, part distressing. PMID- 16220602 TI - Project HOPE. Taking healthcare around the globe. PMID- 16220603 TI - Crystal meth recovery. A step-by-step guide. PMID- 16220604 TI - Why HIV drug resistance matters: an overview. PMID- 16220605 TI - Why I ride. Ride for AIDS Chicago raises funds, and awareness. PMID- 16220606 TI - Haiti--the intersection of race, poverty and HIV. A doctor describes the impact of an epidemic on his homeland. PMID- 16220607 TI - How I became my doctor's free continuing medical education provider. PMID- 16220608 TI - Double balloon GliaSite in the management of recurrent glioblastoma multiforme. PMID- 16220609 TI - Quantitative trait loci for femoral and lumbar vertebral bone mineral density in C57BL/6J and C3h/HeJ inbred strains of mice. (2001). PMID- 16220610 TI - [Rosacea and Demodex]. PMID- 16220611 TI - Pigs, politics, and poor governance. PMID- 16220612 TI - 20 years ago: The British Homeopathic Journal, July 1985. PMID- 16220614 TI - Heritable integration of kDNA minicircle sequences from Trypanosoma cruzi into the avian genome: insights into human Chagas disease. PMID- 16220613 TI - Discretionary death: conditions, dilemmas, and normative regulation. AB - The author examines a major shift in the conceptualization and practices relating to death and dying in Western and other societies with advanced medicine. This shift is the result of socio-technical and cultural developments characterized by but not limited to the routine widespread application of life support technologies in the hospital together with notions of increased patient rights. It has resulted in a class of end-of-life situations, which the author defines as "discretionary death." The concept of discretionary death underscores the role of contextual and discretionary factors in end-of-life decision-making. The author identifies and discusses the necessary and complex process of norm formation that informs and regulates end-of-life medical practice and establishes societal consensus across society with respect to legitimizing "discretionary death." PMID- 16220615 TI - Rethinking the role of older workers: promoting older worker employment in Europe and Japan. PMID- 16220616 TI - Psychological and religious coping strategies of mothers bereaved by the sudden death of a child. AB - The authors examined the associations of 3 types of psychological coping (task based, emotion-based, avoidance), 2 types of religious coping (positive, negative), and their interactions with grief of 57 mothers bereaved by the sudden death of a child. Results indicated that mothers who use emotion-based coping report significantly higher levels of grief, whereas mothers who use avoidance coping report lower levels of grief. The interaction of task coping and positive religious coping was also associated with lower self-reported grief. The findings support the differential utility of various coping styles on mothers' grief reactions to the sudden death of a child. PMID- 16220617 TI - Developmental and contextual correlates of elders' anticipated end-of-life treatment decisions. AB - The purpose of this study was to gain insight into developmental and contextual correlates of the aggressiveness in treatment that community-based elders anticipate they will desire at the end of life. Elders completed questionnaires to measure 4 developmental factors (integrated moral reasoning, self transcendence, past experience with life-threatening illness, and age) and five contextual factors (education level, gender, ethnicity, current health status, and completion of a living will). The variance in desired aggressiveness in treatment was explained by both developmental and contextual correlates. Integrated moral reasoning emerged as a new variable to consider in gaining understanding of the process of end-of-life decisions. PMID- 16220618 TI - Suicide prevention: critical elements for managing suicidal clients and counselor liability without the use of a no-suicide contract. AB - Despite its entrenchment as a standard of practice, no-suicide contracts fail to achieve their purpose as an effective part of treatment or as an effective method of inoculating counselors against potential lawsuits should a client commit suicide. Critical elements for managing suicidal clients and counselor liability without reliance on the no-suicide contract are presented. Therapeutic considerations include evaluating clients to determine (a) no referral for hospitalization needed, (b) referral for voluntary hospitalization, or (c) referral for involuntary hospitalization. PMID- 16220619 TI - The legality of the use of psychiatric neuroimaging in intelligence interrogation. PMID- 16220620 TI - A personal exploration of the power of poetry in palliative care, loss and bereavement. PMID- 16220622 TI - Balancing margin and mission: hospitals alter billing and collection practices for uninsured patients. AB - A barrage of publicity about aggressive hospital billing and collection practices and a spate of lawsuits alleging hospitals overcharged uninsured patients have put hospitals in a harsh national spotlight. In the wake of a campaign by hospital associations to encourage hospitals to create formal policies for billing uninsured patients, many hospitals have modified billing and collection practices for low-income, uninsured patients, according to the Center for Studying Health System Change's (HSC) 2005 site visits to 12 nationally representative communities. Almost all of the hospitals interviewed that had adopted more generous charity care policies indicated expenses previously classified as bad debt have shifted to charity care write-offs. To date, these changes have had little impact on hospital bottom lines, and the impact on access to care for uninsured people remains unclear. PMID- 16220621 TI - Research on environmental health interventions: ethical problems and solutions. AB - This article reviews a variety of ethical issues one must consider when conducting research on environmental health interventions on human subjects. The paper uses the Kennedy Krieger Institute lead abatement study as well as a hypothetical asthma study to discuss questions concerning benefits and risks, risk minimization, safety monitoring, the duty to warn, the duty to report, the use of control groups, informed consent, equitable subject selection, privacy, conflicts of interest, and community consultation. Research on environmental health interventions can make an important contribution to our understanding of human health and disease prevention, provided it is conducted in a manner that meets prevailing scientific, ethical, and legal standards for research on human subjects. PMID- 16220624 TI - Haunted manuscripts: ghost authorship in the medical literature. AB - Ghost authorship occurs when an individual who contributed substantially to a manuscript is not named in the byline or acknowledgments. Ghost authors may be employed by industry to prepare clinical trial results for publication. An expert is then "hired" as author so as to lend an air of credibility and neutrality to the manuscript. Ghost authorship is difficult to detect, and most articles that have been identified as ghostwritten were revealed as such only after investigative work by lawyers, journalists, or scientists. Ghost authorship is ethically questionable in that it may be used to mask conflicts of interest with industry. As it has been demonstrated that industry sponsorship of clinical trials may be associated with outcomes favorable to industry, this is problematic. Evidence-based medicine requires that clinical decisions be based on empirical evidence published in peer-reviewed medical journals. If physicians base their decisions on dubious research data, this can have negative consequences for patients. Ghost authorship also compromises academic integrity. A "film credit" concept of authority is one solution to the problems posed by ghost authorship. Other approaches have been taken by the United Kingdom and Denmark. A solution is necessary, as the relationship between authorship and accountability must be maintained. PMID- 16220623 TI - Inhibition of chronic rejection and development of tolerogenic T cells after ICOS ICOSL and CD40-CD40L co-stimulation blockade. AB - BACKGROUND: Blockade of the CD40-CD40L pathway results in long-term allograft survival but does not prevent chronic rejection. ICOS-ICOSL are members of the CD28-B7 family that play an important role in T-cell activation. METHODS: The authors analyzed the effect of single or combined treatment with an anti-ICOS monoclonal antibody and the fusion molecule CD40 immunoglobulin (Ig) on acute and chronic rejection of heart allografts in rats. RESULTS: Treatment with anti-ICOS resulted in a modest but significant prolongation of allograft survival. Treatment with CD40Ig resulted in long-term graft survival but the cardiac grafts developed chronic rejection lesions. Combined CD40Ig+anti-ICOS treatment led to indefinite graft survival in all recipients and a significant decrease of chronic rejection lesions compared with CD40Ig alone. Importantly, four of the seven CD40Ig+anti-ICOS-treated recipients showed a complete absence of chronic rejection lesions, whereas all of the CD40Ig-treated recipients showed chronic rejection. The CD40Ig+anti-ICOS group also showed significant decreased graft infiltration, decreased antidonor cytotoxic T-lymphocyte activity, and decreased alloantibodies compared with the CD40Ig-treated group. Adoptive transfer of splenocytes indefinitely prolonged allograft survival, whereas those depleted of T cells did not, suggesting the development of T-regulatory mechanisms. CONCLUSIONS. These data indicate that the chronic rejection mechanisms that are CD40-CD40L independent are ICOS-ICOSL dependent. These results were obtained with conservation of cognate immune responses and development of tolerogenic T cells. PMID- 16220625 TI - Reactions to research participation in vulnerable subgroups. AB - This paper describes the extent to which vulnerable individuals (defined by economic, social, psychological, physical health, and child maltreatment status) react to research participation. As part of an ongoing longitudinal study, participants (N=896) completed a lengthy and intrusive in-person interview and provided a small amount of blood through finger pricks. At the end of the interview, participants were asked eight questions about their reactions to the research experience. Vulnerable individuals in general agreed more strongly about having an emotional reaction, but were not less willing to continue to participate. In addition, psychologically vulnerable individuals more strongly agreed they would continue to participate, were treated with respect and dignity, and found their participation meaningful. Compared to whites, nonwhites reported stronger agreement about the meaningfulness of the research and the belief that their responses would be kept private. Like others, individuals vulnerable by virtue of their prisoner status or homelessness (past or current) agreed more strongly about having an emotional reaction to the interview, but otherwise did not differ in their reactions. These results suggest that researchers and institutional review boards should not be deterred from conducting research on sensitive topics with potentially vulnerable populations. PMID- 16220626 TI - Ten steps to developing a national agenda to address financial conflicts of interest in industry sponsored clinical research. AB - Financial liaisons between clinical researchers, research institutions, and industrial sponsors have gained momentum in recent years. In the process, it has been argued by many that trust in the research infrastructure is being eroded by the financial conflicts of interest that emerge from these arrangements. Yet, the financial resources of industry are needed to continue technology transfer from the bench to the bedside. Policy makers and government regulators are currently struggling to determine how to best manage financial conflicts of interest that emerge from these liaisons. Various organizations and government entities have proposed different strategies. This paper explores the limitations of existing measures and recommends that a unified national agenda is needed. We propose 10 steps to develop an agenda to address financial conflicts of interest in industry sponsored clinical research. PMID- 16220627 TI - Medical research, risk, and bystanders. PMID- 16220628 TI - Local IRBs, multicenter trials, and the ethics of internal amendments. PMID- 16220629 TI - The ethics of single blind trials. PMID- 16220631 TI - Liberalism, legal moralism and moral disagreement. AB - According to "legal moralism" it is part of law's proper role to "enforce morality as such". I explore the idea that legal moralism runs afoul of morality itself: there are good moral reasons not to require by law all that there is nevertheless good moral reason to do. I suggest that many such reasons have broad common-sense appeal and could be appreciated even in a society in which everyone completely agreed about what morality requires. But I also critique legal moralism from the special perspective of liberal political justice. Liberalism requires that citizens who disagree with one another on a number of morally significant matters nevertheless coexist and cooperate within a political framework of basic rights protections. When it comes to working out the most basic terms of their political association, citizens are expected to address one another within the limits of what Rawls has called "public reason". Critics of liberalism claim that this is an essentially a-moral (or expedient) attempt to evade substantive moral issues--such as the moral status of the fetus. I argue, on the contrary, that liberalism's emphasis on public reason is itself grounded in very deep--though (suitably) "non-comprehensive"--moral considerations. PMID- 16220630 TI - Respondent burden in clinical research: when are we asking too much of subjects? PMID- 16220632 TI - Do parents lead their children by the hand? AB - The types of gesture+speech combinations children produce during the early stages of language development change over time. This change, in turn, predicts the onset of two-word speech and thus might reflect a cognitive transition that the child is undergoing. An alternative, however, is that the change merely reflects changes in the types of gesture + speech combinations that their caregivers produce. To explore this possibility, we videotaped 40 American child-caregiver dyads in their homes for 90 minutes when the children were 1;2, 1;6, and 1;10. Each gesture was classified according to type (deictic, conventional, representational) and the relation it held to speech (reinforcing, disambiguating, supplementary). Children and their caregivers produced the same types of gestures and in approximately the same distribution. However, the children differed from their caregivers in the way they used gesture in relation to speech. Over time, children produced many more REINFORCING (bike+point at bike), DISAMBIGUATING (that one+ point at bike), and SUPPLEMENTARY combinations (ride+point at bike). In contrast, the frequency and distribution of caregivers' gesture+speech combinations remained constant over time. Thus, the changing relation between gesture and speech observed in the children cannot be traced back to the gestural input the children receive. Rather, it appears to reflect changes in the children's own skills, illustrating once again gesture's ability to shed light on developing cognitive and linguistic processes. PMID- 16220633 TI - Consonant-vowel co-occurrence patterns in Mandarin-learning infants. AB - Most studies on CV co-occurrence patterns in early phonetic development have been based on Indo-European languages. Data from infants learning Mandarin, which has a substantially different phonological system from Indo-European languages, can confirm or refute the findings of previous studies, thus shedding further light on the theoretical bases of CV association. Spontaneous vocalizations were recorded in 45-minute sessions for each of 24 Mandarin-learning infants aged 0;7 to 1;6. In addition, the speech production of 24 caregivers was audio-recorded during their natural daily interactions with the infants at home. Both infants' vocalizations and adults' speech were transcribed and analysed for consonant vowel co-occurrence patterns. These patterns were similar to those in other language groups, but language-specific patterns were evident by 1;0. Combinations of alveolars+front vowels and velars+back vowels confirm Davis & MacNeilage's (1990, 1995) frame-then-content theory and Clements's (1991) model of unified features for consonants and vowels. However, our finding of a language-specific pattern (labials+back vowels) suggests the need to reexamine the 'pure frame' of Davis & MacNeilage and Clements's grouping of labials and rounded vowels. PMID- 16220634 TI - Acquiring the English adjective lexicon: relationships with input properties and adjectival semantic typology. AB - Properties of the input, such as raw frequency and syntactic diversity, have been shown to play a role, to different extents, in the acquisition of nouns and verbs. This study investigated the relationship between three properties of the input (input frequency, syntactic diversity, and variety in noun-type co occurrence) and age of acquisition of English adjectives. In addition, this study explored the relationship between adjectival semantic typology and order of acquisition. The data are 7262 child utterances and 6318 maternal utterances containing one of 272 adjective in the language samples of two children, Sarah and Adam, from age 2;3 to 5;0 (Brown, 1973) available in CHILDES (MacWhinney, 2000). Statistical analyses revealed that all three properties of the input were significantly correlated with age of acquisition, but variety in noun-type co occurrence was not a significant predictor once input frequency and syntactic diversity were taken into account. Semantic analyses revealed that the distribution of adjective types and tokens differed between the children and the mothers (to an extent) when the data were examined in terms of adjectival semantic types. The results suggest that both properties of the input and the semantic properties of the English adjective class play a role in its acquisition. PMID- 16220635 TI - The precocious two-year-old: status of the lexicon and links to the grammar. AB - This is a study of the lexical and grammatical abilities of 16 lexically precocious talkers. These children, aged 2;0 were compared to their age-matched peers, 22 typical talkers aged 2;0, and their expressive vocabulary-matched peers, 22 typical talkers aged 2;6. Individual differences in children's lexical knowledge at 2;0 were stable--evident in parent report, laboratory observation, and an experimental fast-mapping paradigm. In accordance with the continuity hypothesis, the lexically precocious children were also grammatically precocious, having a greater representation of grammatical types and tokens and more advanced combinatorial language than their typical age-matches. Their grammatical development was very similar to that of their older vocabulary-matched peers. Limits on continuity were highly constrained with no true dissociation between the lexicon and the grammar in 33 cases examined. We conclude that, among two year-olds, grammatical development is more tightly associated with the size of the lexicon than with chronological age. PMID- 16220636 TI - The acquisition of auxiliaries BE and HAVE: an elicitation study. AB - Auxiliary syntax is recognized to be one of the more complex aspects of language that children must acquire. However, there is much disagreement among researchers concerning children's early understanding of auxiliaries, with some researchers advocating a relatively abstract grammar as the basis for auxiliary syntax, while others view the acquisition of auxiliary syntax as the gradual accumulation of linguistic knowledge, initially tied to individual lexical items. To investigate the status of children's early knowledge of auxiliary syntax, two studies were carried out. In study 1, 28 children (M= 3;1) were tested for their use of the auxiliaries BE and HAVE in declaratives, while in study 2, 19 children (M=3;3) were tested for their use of these auxiliaries in questions. Although overall error rates were low, there were differences between BE and HAVE in the proportion and types of errors observed in declaratives and questions, and some individual children showed very high error rates. The implications of these findings for different models of auxiliary syntax in children's early utterances are discussed. PMID- 16220637 TI - When opportunity knocks twice: two-year-olds' repetition of sentence subjects. AB - Why are young children's utterances short? This elicited imitation study used a new task--double imitation--to investigate the factors that contribute to children's failure to lexicalize sentence subjects. Two-year-olds heard a triad of sentences singly and attempted to imitate each; they then again heard the same triad singly and again attempted to imitate each. Comparisons between the two attempts showed that children's second passes were more accurate than their first. In addition, independent of sentence length, children increased their inclusion of pronominal and expletive but not lexical subjects. Children included verbs more often from sentences with pronominal than lexical subjects, suggesting a trade-off. Children included subjects more often in short sentences than long ones, and increased subject inclusion only in short sentences. The results suggest that children's language production is similar to adults': a complex interaction of syntactic knowledge, limited cognitive resources, communicative goals, and conversational structure. PMID- 16220638 TI - Negative input for grammatical errors: effects after a lag of 12 weeks. AB - Effects of negative input for 13 categories of grammatical error were assessed in a longitudinal study of naturalistic adult-child discourse. Two-hour samples of conversational interaction were obtained at two points in time, separated by a lag of 12 weeks, for 12 children (mean age 2;0 at the start). The data were interpreted within the framework offered by Saxton's (1997, 2000) contrast theory of negative input. Corrective input was associated with subsequent improvements in the grammaticality of child speech for three of the target structures. No effects were found for two forms of positive input: non-contingent models, where the adult produces target structures in non-error-contingent contexts; and contingent models, where grammatical forms follow grammatical child usages. The findings lend support to the view that, in some cases at least, the structure of adult-child discourse yields information on the bounds of grammaticality for the language-learning child. PMID- 16220639 TI - Parent-child picture-book reading, mothers' mental state language and children's theory of mind. AB - This study focuses on parent-child book reading and its connection to the development of a theory of mind. First, parents were asked to report about frequency of parent-child storybook reading at home. Second, mothers were asked to read four picture-books to thirty-four children between 4;0 and 5;0. Both frequency of parent-child storybook reading at home, and mother's use of mental state terms in picture-books reading tasks were significantly associated with success on false belief tasks, after partialling out a number of potential mediators such as age of children, verbal IQ, paternal education, and words used by mothers in joint picture-book reading. Among the different mental state references (cognitive terms, desires, emotions and perceptions), it was found that the frequency and variety of cognitive terms, but also the frequency of emotional terms correlated positively with children's false belief performance. Relationships between mental state language and theory of mind are discussed. PMID- 16220640 TI - Twin pregnancies: eating for three? Maternal nutrition update. AB - The incidence of multifetal pregnancies has increased, mainly because of assisted reproduction treatments. This trend is reflected in increased maternal and neonatal morbidity and mortality. While the optimum maternal nutrition and weight gain patterns for singleton pregnancies is well documented, there is a paucity of information for twin pregnancies. Although it is assumed that optimum nutritional requirements and weight gains would be greater for twin than for singleton gestations, research is needed to establish the optima. This article is a collation of available recommendations for maternal nutrition and weight gain patterns in twin pregnancies. PMID- 16220641 TI - The protective role of fruits and vegetables against radiation-induced cancer. AB - The role of fruits and vegetables in protecting against radiation-induced cancer and the positive evidence from epidemiology and ancillary investigations are discussed in this review. The recently reported Hiroshima and Nagasaki atomic bomb survivor studies provide strong evidence for the protective role of fruits and vegetables against radiation-induced cancer. The anticarcinogenic substances contained in, and the anticarcinogenic mechanisms proposed for, fruits and vegetables are reviewed. The anticarcinogenic effectiveness of fruits and vegetables are compared with that of dietary supplements. The reasons for the observed superiority of fruits and vegetables are advanced. PMID- 16220642 TI - A Mediterranean-style diet and metabolic syndrome. AB - A moderate increase in physical activity and a detailed and tailored Mediterranean-style diet reduce the prevalence of metabolic syndrome and associated cardiovascular risks through reducing systemic vascular inflammation and endothelium dysfunction, particularly in those patients who do not lose weight. PMID- 16220643 TI - Is GAD2 on chromosome 10p12 a potential candidate gene for morbid obesity? AB - Morbidly obese individuals represent one of the fastest growing subpopulations of obese individuals. Thus, it is of significant interest to broaden our understanding of the potential genetic causes of this public health concern. A recent study investigated a role of positional candidate gene GAD2 (the gene for glutamic acid decarboxylase) in the development of morbid obesity. This commentary carefully examines the genetic and functional arguments for and against the GAD2 gene as an influential gene for obesity. Also discussed are additional research questions that merit inquiry when further evaluating this genetic variant as a putative contributor to human obesity. PMID- 16220644 TI - Developing ourselves. PMID- 16220645 TI - Being a supportive presence in online courses: attending to students' online presence with each other. AB - As a teacher involved in distance education, being a supportive presence includes attending to and facilitating students' knowing and connecting with other. This creates places for learning that are safe, fair, and respectful, which reflects the concerns of the interpretive pedagogies (Diekelmann & Diekelmann, in press). Reflecting a larger issue is the importance of contemporary nurses' knowing and connecting with other team members, and with patients, online in ways that engender community and achieve desired outcomes. With this in mind, attending to the Concernful Practices that enable students to know and connect in ways other than face-to-face encounters becomes even more relevant in preparing for today's practice environments. PMID- 16220646 TI - Doctor of nursing practice program development: reengineering health care. AB - In this article, we describe the developmental process of a Doctor of Nursing Practice (DNP) program that uses interdisciplinary resources to create unique DNP curriculum opportunities. Other schools may benefit from this experience in the development of their own DNP programs. The program delivers an innovative curriculum from post-baccalaureate to doctorate, emphasizing health care engineering and interdisciplinary collaboration among faculty, hospitals, community leaders, and policymakers. This DNP program is uniquely situated to provide leadership in solving complex clinical problems through its partnership with the Regenstrief Center for Healthcare Engineering, the School of Pharmacy, the Homeland Security Institute, and the Center on Aging and the Life Course. Doctoral coursework, interdisciplinary collaboration, health care engineering/systems approaches, and new knowledge result in uniquely qualified providers. Post-baccalaureate students complete the university's Adult Nurse Practitioner program or its developing Pediatric Nurse Practitioner program during the first 2 years of the 4-year curriculum. A total of 83 post baccalaureate credit hours include 1,526 hours of supervised clinical practice, a health policy residency, and cognate residencies in an area of specialization. The seven core competencies recommended by the American Association of Colleges of Nursing are incorporated into the curriculum. PMID- 16220648 TI - Hermeneutic notions augment cultural safety education. AB - Nurses are beginning to demand educational approaches that confront racism, rather than teach cultural diversity. One example of the latter approach is the introduction of kawa whakaruruhau, or cultural safety, in nursing and midwifery education in New Zealand. In the nursing and midwifery context of kawa whakaruruhau, nurses and midwives recognize, respect, and nurture the unique cultural identity of New Zealand's indigenous people, the tangata whenua, and safely meets their needs, expectations, and rights. In this article, I integrate literature pertaining to the implementation of cultural safety with the findings of a hermeneutic project that described the experience of nursing people from cultures other than one's own, and argue that the Gadamerian notions of "horizon," "prejudice," and "play" can be used to facilitate understanding of the tensions and contradictions inherent in cross-cultural practice. In addition, I recommend strategies that enable students to explore the prejudices, paradoxes, and possibilities experienced personally and professionally. As Gadamer noted, the art is in seeing what is questionable. There is also art in knowing how to question in a manner that makes new understanding possible. PMID- 16220647 TI - Social justice as a framework for professional nursing. AB - A persistent connection exists between health disparity and societal inequality. Since more research is designed to document, rather than alter, those trends, nurses are called on to reinvest in a social justice agenda. An educational focus on social justice is necessary to prepare future nurses to address health concerns related to how societies are structured. This article reports on how social justice was used as a framework to teach concepts related to professional nursing. The course structure is described, a focus on how the course content was taught is outlined, and conclusions are drawn. Linking content on social justice to professional nursing is important because research indicates that, if societal relationships are more equal, population health indicators between diverse groups become more stable nationally and globally. PMID- 16220649 TI - The effect of a training course on nursing students' attitudes toward, perceptions of, and confidence in managing patient aggression. AB - Patient aggression is a problem in many health care settings, and nursing students are among the most vulnerable to experiencing such aggression. Training courses have been suggested to help nurses better manage patient aggression. Such courses can lead to changes in attitudes toward and perceptions of, as well as greater confidence in managing, aggression. In this quasi-experimental study, we investigated the effect of a training course on nursing students' attitudes toward, perceptions of, and confidence in managing patient aggression. Students in the intervention group demonstrated enhanced confidence but no change in attitude after the training course, while students in the control group remained stable on all measures. The short time frame of the study, the training course itself, and the instruments used for monitoring attitudes toward aggression are possible reasons for these results. We tentatively conclude that it is possible to enhance nursing students' perceived confidence in managing patient aggression without changing their fundamental views of it. PMID- 16220650 TI - Human patient simulators: a new face in baccalaureate nursing education at Brigham Young University. AB - Simulation has been used to augment learning in the health care professions. The human patient simulator (HPS) has been used primarily by medical students but is now being used by nursing students, as well. This study explored the benefits and limitations of using an HPS as a patient substitute for one day of actual clinical experience for junior nursing students. Learning outcomes included increased student knowledge, ability, and confidence in medication administration. PMID- 16220651 TI - Nurse scholars program. AB - The Nurse Scholars Program was developed for two reasons: to attract highly qualified students to nursing, and to increase the retention of graduates as they enter employment in local agencies following graduation. Scholars receive dollar 10,000 for each of their junior and senior years and are asked to work for 1 year at the agency for each dollar 10,000 received. Additional monies are set aside to assist graduate and doctoral students through stipends. Scholars are paired with Mentors, nurses selected for their expertise and experience and brought into the "family" of the hospital through special events. Mentors are included in educational activities and related programs. To date, dollar 840,000 has been committed to the program, and data regarding the success of the program are continually being assessed. PMID- 16220652 TI - The breastfeeding case study: a model for educating nursing students. AB - Due to several national initiatives and recommendations from professional organizations, more women may be encouraged to breastfeed. However, they will not achieve improvement in breastfeeding duration if their health care providers lack knowledge about breastfeeding or are not easily accessible in some areas. Nurses who work with families with children are responsible for a great deal of patient education and can affect women's decisions to initiate and continue breastfeeding. Therefore, it is critical that nurse educators find a way to address this knowledge gap, especially in already overburdened curricula. This article describes a successful model for doing so. PMID- 16220653 TI - Entering a brave new world: an interdisciplinary health care course on weapons of mass destruction. PMID- 16220654 TI - Using a collaborative learning method to enhance mastery of pathophysiology content. PMID- 16220655 TI - [Mitochondrial dysfunction in bipolar disorder]. AB - Phosphorus magnetic resonance spectroscopic studies in bipolar disorder revealed altered brain energy metabolism resembling that of chronic progressive external ophthalmoplegia (CPEO). Mood disorder is one characteristic symptom in several families of CPEO caused by mutations of three genes, ANT1, Twinkle, and POLG. Molecular genetic analysis revealed association of bipolar disorder with mitochondrial DNA (mtDNA) 10398A polymorphism, 3644C mutation, and FDUFV2. In the postmortem brains, increased levels of mtDNA 4977bp deletion and 3243G mutation, and altered expression of mitochondria-related genes were reported. Mitochondria play an important role in neuroplasticity and apoptotic signaling via regulating intracellular calcium homeostasis. Thus, mitochondrial dysfunction may cause altered calcium homeostasis and neuroplasticity, resulting in bipolar disorder. Most molecular genetic findings in bipolar disorder regarding mitochondria and endoplasmic reticulum stress signaling are common to Parkinson's disease and diabetes mellitus. Thus, it is possible that bipolar disorder is also a disease caused by the progressive loss of some neuronal cells. PMID- 16220656 TI - [Pharmacology of an omega receptor agonist]. AB - It has been suggested that different BZP (omega) receptor subtypes may mediate distinct behavioral effects of BZP receptor ligands. Several studies demonstrated that omega1 selective compounds are characterized by an increase in slow wave deep sleep with rapid onset of hypnotic action, and by reduced muscle relaxation, amnesic liability or tolerance. On the other hand, it is known that many different subtypes of GABAA receptors exist, based on the fact that many different subtypes can go into assembling GABAA receptors. GABAA receptors with alpha1 subunits may mediate sedative action and perhaps amnesia. Those with alpha2 subunits probably mediate anxiolytic actions. GABAA receptors with alpha3 subunits may regulate various neurotransmitters, and those with alpha5 subunits may also contribute to amnesia. Such discoveries could open up new avenues for drug development. PMID- 16220657 TI - [Chromosome 22q11 and schizophrenia]. AB - Several human chromosomal regions have been identified as candidate regions that play a role in schizophrenia. Deletion or duplication of chromosome 22q11 is associated with velo-cardio-facial syndrome/DiGeorge syndrome (VCFS/DGS), a disorder associated with high rates of schizophrenia as well as physical abnormalities (i.e., cardiovascular, parathyroid, thymic and craniofacial abnormalities). Recent mouse studies have identified several candidate genes for VCFS/DGS within the mouse homologue chromosome 16. Deletion of Tbx1, Prodh and Comt within mouse chromosome 16 causes several physical and behavioral features of VCFS/DGS. As VCFS/DGS is likely to represent a genetic subtype of schizophrenia, pinpointing the genetic basis for this specific subtype will contribute to a better understanding of this neuropsychiatric disorder. PMID- 16220658 TI - [Effect of Ginkgo biloba extract on memory deficits in radial maze performance induced by some drugs in rats]. AB - Ginkgo biloba extract is widely used as a herbal medicine or dietary supplement in Europe, since Ginkgo biloba extract is effective in facilitation of learning and recollection of memories. However, little is known about the mechanism of the action of Ginkgo biloba extract on learning and memory enhancements. On the other hand, it is well known that cholinergic, histaminergic and glutamatergic systems play a crucial role in learning and memory in animals. Therefore, in order to elucidate the mechanism of Ginkgo biloba extract on memory, we studied and clarified the effect of Ginkgo biloba extract on spatial memory deficits induced by scopolamine, diphenhydramine or MK-801 using eight-arm radial maze performance. It was found that Ginkgo biloba extract improved the spatial memory deficits induced by scopolamine. Ginkgo biloba extract also caused repair to spatial memory deficits induced by diphenhydramine. On the other hand, no significant effect was observed with MK-801-induced spatial memory deficits. These findings suggest that the effect of Ginkgo biloba extract is mediated not only by the cholinergic system but also by the histaminergic system to induce learning and memory enhancements. PMID- 16220659 TI - [The functional change in the 5-HT1A receptor induced by stress and the role of the 5-HT1A receptor in neuroprotection]. AB - Mice exposed to various stresses, especially restrained-stress, revealed the anxiogenic effect detected by the light-dark test. Under this condition, a remarkable decrease in [35S]GTPgammaS binding to membranes from the prefrontal cortex, amygdala and hypothalamus of restrained-stress mice stimulated by the selective 5-HT1A receptor agonist 5-carboxamidotriptamine (5-CT) was clearly observed, whereas a significant increase in [35S]GTPgammaS binding stimulated by the 5-HT1A receptor agonist was clearly observed in the dorsal raphe nuclei (DRN) of restrained-stress mice. The immunohistochemical study showed a drastic reduction in phosphorylated-CREB-like immunoreactivity in the DRN of restrained stress mice. Furthermore, we found a drastic reduction in myelin-associated glycoprotein (MAG)-like immunoreactivity (MAG-IR) in the DRN, amygdala and hypothalamus, indicating the direct suppression of synaptic transmission in these regions. It has been accepted that GSK3beta in the Wnt signal pathway plays an important role in various neuronal functions including apoptosis, clustering of synapsin I and early growth and axonal remodeling. In the present study, the increase in protein levels of GSK3beta and phosphorylated-GSK3beta to cytosol fractions of the amygdala was noted in restrained-stress mice. Taken together, these results suggest that restrained stress may directly affect the 5-HT1A receptor-regulated synaptic transmission in the brain, leading to the expression of the anxiogenic effect in mice. It is well known that various stresses induce intracellular oxidative stress. The present study was then undertaken to investigate the effect of the stimulation of 5-HT1A receptors on oxidative stress. Treatment with H2O2 caused the activation of caspase-3-positive cells and the reduction in levels of MAG-IR in the limbic neuron/glia cocultures as compared to medium alone. The stimulation of 5-HT1A receptor by 5-CT produced a dramatic protection against H2O2-triggered activation of caspase-3 and reduction in levels of MAG-IR. These results suggest that 5-HT1A receptors were involved in the modulation of anxiety and the understanding of molecular mechanisms of 5-HT1A receptor-related cascades may pave the way for new therapeutic strategies for affective disorders. PMID- 16220660 TI - [A mechanism for modulation of N-methyl-D-aspartate receptor channels by free ferrous ions in cultured rat cortical neurons]. AB - Rat cortical neurons cultured in DMEM/F-12 in the absence of fetal calf serum were harvested on days 3, 6 and 9 in vitro (DIV), followed by homogenization and subsequent SDS-PAGE for immunoblotting using an antibody against MAP-2 or GFAP. Expression of MAP-2 was not markedly changed during cultivation for 3 to 9 DIV, while GFAP was profoundly expressed after 6 DIV in a manner dependent on the duration of culturing. Cortical neurons cultured for 3 DIV were pre-incubated with fluo-3 AM and then subjected to fluorescence image analysis using a confocal microscope. The exposure to NMDA markedly increased the number of neurons with high fluorescence intensity in the absence of MgCl2, without prominently affecting that in the presence of MgCl2. The addition of FeCI2, but not hemoglobin and transferrin, markedly reduced the increase in a concentration dependent manner in the presence of NMDA. This inhibition by FeCl2 was not affected by the addition of dithiothreitol or 2-mercaptoethanol. These results suggest that free ferrous ions may selectively prevent Ca2+ influx across NMDA receptor channels in a manner different from that done by the redox site on the channels in cultured rat cortical neurons. PMID- 16220661 TI - AVMA mounts preparedness, response to Katrina. PMID- 16220662 TI - U.S. plans to modernize Plum Island facility. PMID- 16220663 TI - Dr. Ramey remains unconvinced about acupuncture. PMID- 16220664 TI - Postmarketing surveillance for dog and cat vaccines: new resources in changing times. PMID- 16220665 TI - What practitioners should know about bovine spongiform encephalopathy to serve clients and the cattle industry. PMID- 16220667 TI - Employment, starting salaries, and educational indebtedness of year-2005 graduates of US veterinary medical colleges. PMID- 16220666 TI - What is your diagnosis? Inflammatory and infiltrative disease of the colon. PMID- 16220668 TI - Comparison of arthroscopic and radiographic abnormalities in the hip joints of juvenile dogs with hip dysplasia. AB - OBJECTIVE: To compare radiographic and arthroscopic abnormalities in juvenile dogs with clinically apparent hip dysplasia. DESIGN: Case series. ANIMALS: 52 dogs (70 hip joints) with clinical signs of hip dysplasia scheduled to undergo triple pelvic osteotomy. PROCEDURE: A ventrodorsal radiographic projection of the pelvis was evaluated by a radiologist unaware of clinical and arthroscopic findings, and radiographic osteoarthritic abnormalities were judged and scored as absent (0), mild (1), moderate (2), or severe (3). Arthroscopy was performed by a surgeon unaware of clinical and radiographic findings, and arthroscopic abnormalities were graded from 0 (normal) to 5 (exposed, eburnated subchondral bone). RESULTS: In 30 of the 70 (43%) hip joints, no radiographic osteoarthritic abnormalities were seen. Severe, full-thickness articular cartilage lesions (grade 4) of the femoral head or acetabulum were seen arthroscopically in 14 (20%) joints. Lesions > or = grade 2 were seen in 60 (86%) joints. Partial tearing of the ligament of the femoral head was present in 57 (81%) joints, and complete rupture was seen in 5 (7%). Radiographic abnormalities were seen in 13 of the 14 (93%; 95% confidence interval, 66% to 99.8%) joints with grade 4 arthroscopic abnormalities but in only 23 of the 46 (50%; 95% confidence interval, 35% to 65%) joints with grade 2 or 3 arthroscopic abnormalities. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that radiography is not a sensitive method for identifying moderate cartilage lesions in juvenile dogs with hip dysplasia. If moderate cartilage lesions are an important prognostic indicator for the success of triple pelvic osteotomy, then methods other than radiography should be used to detect these lesions. PMID- 16220669 TI - Evaluation of serum 17-hydroxyprogesterone concentration after administration of ACTH in dogs with hyperadrenocorticism. AB - OBJECTIVE: To evaluate serum 17-hydroxyprogesterone (17-OHP) concentration measurement after administration of ACTH for use in the diagnosis of hyperadrenocorticism in dogs. DESIGN: Prospective study. ANIMALS: 110 dogs. PROCEDURE: Serum 17-OHP concentrations were measured before and after ACTH stimulation in 53 healthy dogs to establish reference values for this study. Affected dogs had pituitary-dependent (n = 40) or adrenal tumor-associated (12) hyperadrenocorticism or potentially had atypical hyperadrenocorticism (5; diagnosis confirmed in 1 dog). In affected dogs, frequency interval and borderline and abnormal serum 17-OHP concentrations after ACTH stimulation were determined. Serum cortisol concentrations were assessed via low-dose dexamethasone suppression and ACTH stimulation tests. RESULTS: In healthy dogs, serum 17-OHP concentration frequency intervals were grouped by sex and reproductive status (defined as < 95th percentile). Frequency intervals of serum 17-OHP concentrations after ACTH stimulation were < 77, < 2.0, < 3.2, and < 3.4 ng/mL (< 23.3, < 6.1, < 9.7, and < 10.3 nmol/L) for sexually intact and neutered females and sexually intact and neutered males, respectively. In 53 dogs with confirmed hyperadrenocorticism, serum cortisol concentrations after ACTH stimulation and 8 hours after administration of dexamethasone and serum 17-OHP concentrations after ACTH stimulation were considered borderline or abnormal in 79%, 93%, and 69% of dogs, respectively. Two of 5 dogs considered to have atypical hyperadrenocorticism had abnormal serum 17-OHP concentrations after ACTH stimulation. CONCLUSIONS AND CLINICAL RELEVANCE: Serum 17-OHP concentration measurement after ACTH stimulation may be useful in the diagnosis of hyperadrenocorticism in dogs when other test results are equivocal. PMID- 16220670 TI - Adverse events diagnosed within three days of vaccine administration in dogs. AB - OBJECTIVE: To determine incidence rates and potential risk factors for vaccine associated adverse events (VAAEs) diagnosed within 3 days of administration in dogs. DESIGN: Retrospective cohort study. ANIMALS: 1,226,159 dogs vaccinated at 360 veterinary hospitals. PROCEDURE: Electronic records from January 1, 2002, through December 31, 2003, were searched for possible VAAEs (nonspecific vaccine reaction, allergic reaction, urticaria, or anaphylaxis) diagnosed within 3 days of vaccine administration. Information included age, weight, sex, neuter status, and breed. Specific clinical signs and treatments were reviewed in a random sample of 400 affected dogs. The association between potential risk factors and a VAAE was estimated by use of multivariate logistic regression. RESULTS: 4,678 adverse events (38.2/10,000 dogs vaccinated) were associated with administration of 3,439,576 doses of vaccine to 1,226,159 dogs. The VAAE rate decreased significantly as body weight increased. Risk was 27% to 38% greater for neutered versus sexually intact dogs and 35% to 64% greater for dogs approximately 1 to 3 years old versus 2 to 9 months old. The risk of a VAAE significantly increased as the number of vaccine doses administered per office visit increased; each additional vaccine significantly increased risk of an adverse event by 27% in dogs < or = 10 kg (22 lb) and 12% in dogs > 10 kg. CONCLUSIONS AND CLINICAL RELEVANCE: Young adult small-breed neutered dogs that received multiple vaccines per office visit were at greatest risk of a VAAE within 72 hours after vaccination. These factors should be considered in risk assessment and risk communication with clients regarding vaccination. PMID- 16220671 TI - Prevalence of cranial cruciate ligament rupture in a population of dogs with lameness previously attributed to hip dysplasia: 369 cases (1994-2003). AB - OBJECTIVE: To determine the prevalence of cranial cruciate ligament rupture (CCLR) in dogs with lameness previously attributed to canine hip dysplasia (CHD). DESIGN: Retrospective study. ANIMALS: 369 client-owned dogs. PROCEDURES: Hospital medical records from 1994 to 2003 were reviewed for dogs in which the referring veterinarian had diagnosed hip dysplasia or hip pain. Dogs were designated as having hind limb lameness because of partial or complete CCLR or CHD. RESULTS: 8% of dogs were sexually intact females, 43% were spayed females, 14% were sexually intact males, and 35% were castrated males. Mean age was 3.8 years (range, 3 months to 15 years). The most common breeds were the Labrador Retriever (21%), German Shepherd Dog (13%), and Golden Retriever (11%). The prevalence of CCLR as the cause of hind limb lameness was 32% (95% confidence interval, 27.2% to 36.8%). The distribution of CCLR among hind limbs was left (29%), right (28%), and bilateral (43%). Of 119 dogs with CCLR, 94% had concurrent radiographic signs of CHD, 92% had stifle joint effusion, and 81% had a cranial drawer sign. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of the high prevalence of CCLR in dogs referred for lameness because of CHD, it is important to exclude other sources of stifle joint disease before making recommendations for treatment of CHD. PMID- 16220672 TI - Gastrointestinal tract perforation in dogs treated with a selective cyclooxygenase-2 inhibitor: 29 cases (2002-2003). AB - OBJECTIVE: To identify factors associated with gastrointestinal tract perforation in dogs being treated with a selective cyclooxygenase-2 (COX-2) inhibitor (deracoxib). DESIGN: Retrospective study. ANIMALS: 29 dogs. PROCEDURE: The Novartis Animal Health pharmacovigilance database was searched for records of dogs treated with deracoxib in which gastrointestinal tract perforation was documented. Results-16 of the 29 (55%) dogs had received deracoxib at a dosage higher than that approved by the FDA for the particular indication being treated, with 25 (86%) dogs having received deracoxib at a dosage > 2 mg/kg/d (0.9 mg/lb/d). Seventeen (59%) dogs had received at least 1 other nonsteroidal anti inflammatory drug (NSAID) or a corticosteroid in close temporal association (within 24 hours) with deracoxib administration (ie, immediately before or following). In all, 26 (90%) dogs had received deracoxib at a higher-than approved dosage or had received at least 1 other NSAID or corticosteroid in close temporal association with deracoxib administration. Twenty dogs died or were euthanatized, and 9 survived. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with gastrointestinal tract perforation and that had been treated with deracoxib, perforation was most likely attributable to a number of factors. Deracoxib should only be used at approved dosages. Cortico-steroids and other less selective NSAIDs should not be administered in close temporal association with selective COX-2 inhibitors, including deracoxib. Further study is required to define this problem. PMID- 16220673 TI - Response rates and survival times for cats with lymphoma treated with the University of Wisconsin-Madison chemotherapy protocol: 38 cases (1996-2003). AB - OBJECTIVE: To determine response rates and survival times for cats with lymphoma treated with the University of Wisconsin-Madison chemotherapy protocol. DESIGN: Retrospective study. ANIMALS: 38 cats with lymphoma. PROCEDURE: Medical records were reviewed, and information on age, sex, breed, FeLV and FIV infection status, anatomic form, clinical stage, and survival time was obtained. Immunophenotyping was not performed. RESULTS: Mean +/- SD age of the cats was 10.9 +/- 4.4 years. Overall median survival time was 210 days (interquartile range, 90 to 657 days), and overall duration of first remission was 156 days (interquartile range, 87 to 316 days). Age, sex, anatomic form, and clinical stage were not significantly associated with duration of first remission or survival time. Eighteen of the 38 (47%) cats had complete remission, 14 (37%) had partial remission, and 6 (16%) had no response. Duration of first remission was significantly longer for cats with complete remission (654 days) than for cats with partial remission (114 days). Median survival time for cats with complete remission (654 days) was significantly longer than median survival time for cats with partial remission (122 days) and for cats with no response (11 days). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that a high percentage of cats with lymphoma will respond to treatment with the University of Wisconsin-Madison chemotherapy protocol. Age, sex, anatomic form, and clinical stage were not significantly associated with duration of first response or survival time, but initial response to treatment was. PMID- 16220674 TI - Effect of a 24-hour infusion of an isotonic electrolyte replacement fluid on the renal clearance of electrolytes in healthy neonatal foals. AB - OBJECTIVE: To determine the effects of a 24-hour infusion of an isotonic electrolyte replacement fluid (IERF) on weight, serum and urine electrolyte concentrations, and other clinicopathologic variables in healthy neonatal foals. ANIMALS: 4 healthy 4-day-old foals. DESIGN: Prospective study. PROCEDURE: An IERF was administered to each foal at an estimated rate of 80 mL/kg/d (36.4 mL/lb/d) for 24 hours. Body weight was measured before and after the infusion period. Urine was collected via catheter during 4-hour periods; blood samples were collected at 4-hour intervals. Variables including urine production; urine and serum osmolalities; sodium, potassium, and chloride concentrations in urine and serum; urine and serum creatinine concentrations; urine osmolality-to-serum osmolality ratio (OsmR); transtubular potassium gradient (TTKG); and percentage creatinine clearance (Cr(cl)) of electrolytes were recorded at 0, 4, 8, 12, 16, 20, and 24 hours during the infusion period. Immediately after the study period, net fluid and whole-body electrolyte changes from baseline values were calculated. RESULTS: Compared with baseline values, urine and serum sodium and chloride serum concentrations, urine and serum osmolalities, OsmR, and percentage Cr(cl) of sodium and chloride were significantly increased at various time points during the infusion; urine production did not change significantly. After 24 hours, weight, TTKG, serum creatinine concentration, and whole-body potassium had significantly decreased from baseline values. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of an IERF containing a physiologic concentration of sodium may not be appropriate for use in neonatal foals that require maintenance fluid therapy. PMID- 16220675 TI - Surgical correction of ear curling caused by scar tissue formation in a horse. AB - An 18-month-old Belgian Warmblood mare was evaluated because it had injured the outer convex aspect of the left auricle. Second intention healing of the wound area caused tissue contracture, which resulted in the tip of the ear curling backward. By use of a technique involving undermining of the skin and a flap of granulation tissue on the medial aspect of the wound area and multiple incisions of the auricular cartilage, the curling was relieved and the ear regained a more normal shape. A skin graft was applied to cover the existing wound defect in an attempt to accelerate wound healing; thermoplastic material was contoured to fit the inner concave surface of the ear for immobilization and fixation of the ear in its final shape after surgery. Thirty days after surgery, the graft had healed completely and the ear had a normal conformation. The successful outcome of this treatment suggests that correction of an ear deformity secondary to scar tissue formation by use of an adapted surgical technique and appropriate materials can be achieved with good cosmetic results in horses. PMID- 16220676 TI - Evaluation of risk factors for development of catheter-associated jugular thrombophlebitis in horses: 50 cases (1993-1998). AB - OBJECTIVE: To evaluate risk factors associated with development of catheter associated jugular thrombophlebitis in hospitalized horses. DESIGN: Retrospective case-control study. ANIMALS: 50 horses with thrombophlebitis and 100 control horses. PROCEDURE: Medical records from 1993 through 1998 were searched for horses with thrombophlebitis. Horses that were hospitalized for at least 5 days, had an i.v. catheter placed in a jugular vein (other than for solely anesthetic purposes), and had no evidence of thrombophlebitis during admission or hospitalization were chosen as controls. Signalment, history, clinicopathologic findings, primary illness, and treatment were obtained from the medical records. Data were analyzed by use of logistic regression to perform univariate and multivariate analyses. RESULTS: For a horse with endotoxemia, the odds of developing thrombophlebitis were 18 times those for a similar horse without endotoxemia. For a horse with salmonellosis, the odds of developing thrombophlebitis were 68 times those for a similar horse without salmonellosis. For a horse with hypoproteinemia, the odds of developing thrombophlebitis were almost 5 times those for a similar horse without hypoproteinemia. For a horse in the medicine section, the odds of developing thrombophlebitis were 16 times those for a similar horse in the surgery section. For a horse with large intestinal dise, the odds of developing thrombophlebitis were 4 times those for a similar horse without large intestinal disease. For a horse receiving antidiarrheal or antiulcerative medications, the odds of developing thrombophlebitis were 31 times those for a similar horse not receiving these medications. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that patient factors, including large intestinal disease, hypoproteinemia, salmonellosis, and endotoxemia, were associated with development of catheter-associated thrombophlebitis in horses. PMID- 16220677 TI - One-step laparoscopic abomasopexy for correction of left-sided displacement of the abomasum in dairy cows. AB - Four adult dairy cows in which a diagnosis of left-displaced abomasum (LDA) had been made underwent a 1-step laparoscopic abomasopexy (LA). The technique was performed with each cow positioned in dorsal recumbency. Two laparoscopic portals were created in the right paramedian area to identify the abomasum and direct insertion of the steel trocar and cannula into the abomasal lumen. A stainless steel toggle pin (with 2 lengths of suture attached to its midpoint) was inserted via the cannula into the abomasal lumen while the excess suture material remained exterior to the abdomen. The abomasum was deflated, and the excess suture material was withdrawn up to a preset marker on the suture to position the abomasum adjacent to the body wall. The suture was then tied to secure the abomasum in place. By use of this 1-step LA technique, LDA was successfully corrected in all 4 cows. The procedure is minimally invasive and allows viewing of the abomasum for correct positioning and fixation; it can be accomplished with the speed associated with the blind roll-and-tack technique. The 1-step LA technique may reduce the incidence of complications associated with traditional laparotomy and the blind roll-and-tack technique and could be a useful alternative procedure for the treatment of LDA in dairy cows. PMID- 16220678 TI - Demystifying Medicare's new prescription drug benefit. PMID- 16220679 TI - A job to be proud of. PMID- 16220681 TI - PHA ensures correct site surgery. PMID- 16220682 TI - A case for preserving CON. PMID- 16220680 TI - Not so fast! DOJ speaks on HIPAA criminal liability. PMID- 16220683 TI - Multiple alignment through protein secondary-structure information. AB - It is well known that protein secondary-structure information can help the process of performing multiple alignment, in particular when the amount of similarity among the involved sequences moves toward the "twilight zone" (less than 30% of pairwise similarity). In this paper, a multiple alignment algorithm is presented, explicitly designed for exploiting any available secondary structure information. A layered architecture with two interacting levels has been defined for dealing with both primary- and secondary-structure information of target sequences. Secondary structure (either available or predicted by resorting to a technique based on multiple experts) is used to calculate an initial alignment at the secondary level, to be arranged by locally scoped operators devised to refine the alignment at the primary level. Aimed at evaluating the impact of secondary information on the quality of alignments, in particular alignments with a low degree of similarity, the technique has been implemented and assessed on relevant test cases. PMID- 16220684 TI - JViz.Rna--a Java tool for RNA secondary structure visualization. AB - Many tools have been developed for visualization of RNA secondary structures using a variety of techniques and output formats. However, each tool is typically limited to one or two of the visualization models discussed in this paper, supports only a single file format, and is tied to a specific platform. In order for structure prediction researchers to better understand the results of their algorithms and to enable life science researchers to interpret RNA structure easily, it is helpful to provide them with a flexible and powerful tool.jViz.Rna is a multiplatform visualization tool capable of displaying RNA secondary structures encoded in a variety of file formats. The same structure can be viewed using any of the models supported, including linked graph, circle graph, dot plot, and classical structure. Also, the output is dynamic and can easily be further manipulated by the user. In addition, any of the drawings produced can be saved in either the EPS or PNG file formats enabling easy usage in publications and presentations. PMID- 16220685 TI - P-RnaPredict--a parallel evolutionary algorithm for RNA folding: effects of pseudorandom number quality. AB - This paper presents a fully parallel version of RnaPredict, a genetic algorithm (GA) for RNA secondary structure prediction. The research presented here builds on previous work and examines the impact of three different pseudorandom number generators (PRNGs) on the GA's performance. The three generators tested are the C standard library PRNG RAND, a parallelized multiplicative congruential generator (MCG), and a parallelized Mersenne Twister (MT). A fully parallel version of RnaPredict using the Message Passing Interface (MPI) was implemented on a 128 node Beowulf cluster. The PRNG comparison tests were performed with known structures whose sequences are 118, 122, 468, 543, and 556 nucleotides in length. The effects of the PRNGs are investigated and the predicted structures are compared to known structures. Results indicate that P-RnaPredict demonstrated good prediction accuracy, particularly so for shorter sequences. PMID- 16220686 TI - Multiple SVM-RFE for gene selection in cancer classification with expression data. AB - This paper proposes a new feature selection method that uses a backward elimination procedure similar to that implemented in support vector machine recursive feature elimination (SVM-RFE). Unlike the SVM-RFE method, at each step, the proposed approach computes the feature ranking score from a statistical analysis of weight vectors of multiple linear SVMs trained on subsamples of the original training data. We tested the proposed method on four gene expression datasets for cancer classification. The results show that the proposed feature selection method selects better gene subsets than the original SVM-RFE and improves the classification accuracy. A Gene Ontology-based similarity assessment indicates that the selected subsets are functionally diverse, further validating our gene selection method. This investigation also suggests that, for gene expression-based cancer classification, average test error from multiple partitions of training and test sets can be recommended as a reference of performance quality. PMID- 16220687 TI - Molecular dynamics simulations of the cytolytic toxin Cyt1A in solution. AB - Cytolytic toxin Cyt1A from Bacillus thuringiensis var. israelensis is used as an environmentally friendly insecticide, but its mode of action has not been clearly established. One main obstacle seems to be the lack of the experimentally determined structure of the toxin. As a first step in computer simulations of Cyt1A, in this paper, a three-dimensional molecular structure of Cyt1A in solution was generated by homology modeling, potential energy minimization and molecular dynamics. Regions of the toxin molecule that manifest increased conformational flexibility--and thus are likely to participate in the initial membrane binding and conformational changes--were then identified. Finally, the simulated structure was used to study the effect of a single amino-acid mutation that is known to abrogate the toxicity of Cyt1A in vivo. PMID- 16220688 TI - A computational approach for peptidomic analysis in taxonomic study of amphibian species. AB - Peptides in the skin secretion of frogs have been studied for some time now because they frequently possess important biological activity such as antibiotic, antimicrobial, or anticancer properties. In this paper, we present a computational approach for measuring the degree of similarity between the entire peptide complement of the skin secretion of specimens from the same or different species. The first step in the analysis is the generation of a mass spectral profile from an experimental high-performance liquid chromatography/electrosparay ionization analysis of the sample. An "overlap" between the mass spectral profiles of different specimens is then proposed as a measure of their similarity. Analysis of specimens from three species of the genus Litoria, viz., L. Aurea, L. Caerulea, and L. Infrafrenata, and Rana Capito of genus Rana shows that the degree of similarity is highest between specimens from the same species, lower for specimens from different species of the same genus, and lowest between specimens from different genera. This indicates that comparison of skin peptide profiles (i.e., mass spectral profiles of skin secretion) is potentially a useful aid in the taxonomic study of amphibian species. PMID- 16220689 TI - DNA microarray stochastic model. AB - A stochastic model of the DNA microarray image pixels is presented. The model includes spot pixel intensity distribution, interpixel correlations and the intensity distribution of background noise. The data is indicative of a small exponential additive noise process and a larger Gaussian fluctuation that scales with spot intensity. Correlations are observed among pixels in the spot and between test and control images. The correlated fluctuations may be attributed to variations across each spot in the amount of DNA placed on the spot during the array fabrication process. The model may be used in gene expression estimation algorithm development, both to test new algorithms through simulation and to develop optimum algorithms. The model should also be easily adapted to new array based technologies in proteomics. PMID- 16220691 TI - Effects of combined ozone and air pollution particle exposure in mice. AB - Epidemiologic studies indicate that ozone (O3*) and air pollution particles can exacerbate asthma symptoms. We investigated whether coexposure to inhaled particles and O3 causes a synergistic effect on airway responsiveness and allergic inflammation in a murine (BALB/c) model of ovalbumin (OVA)-induced asthma. Half of the mice were sensitized by intraperitoneal injection of OVA and then exposed to OVA aerosol on 3 successive days to create the asthmatic phenotype; the other half were sensitized to OVA and exposed to phosphate buffered saline (PBS) to create the nonasthmatic control group. On the same 3 days that the OVA or PBS challenge was administered, mice were further divided into groups that were exposed for 5 hours to concentrated ambient particles (CAPs; mass values ranging from 63 to 1,569 microg/m3 for 1 day's exposure), 0.3 ppm O3, both, or neither (n > or = 61 total mice per exposure group for all 12 experiments). Whole-body plethysmography was used to measure airway responsiveness after challenge with aerosolized methacholine (MCh). Enhanced pause (Penh), an index that closely correlates with pulmonary resistance (Hamelmann et al 1997), was measured daily in each mouse immediately after pollutant exposure and, for 7 of the 12 experiments (n > or = 36/exposure group), beginning 24 hours after the final OVA or PBS challenge. Using several complementary statistical models, we found that exposure to CAPs alone caused a small but significant increase in Penh in both normal and asthmatic mice immediately after exposure (an increase of approximately 1% per 100-microg/m3 increase in CAPs). No increase in Penh was found in animals exposed to O3 alone or to filtered air. Compared with control animals, no combination of exposure atmosphere plus asthma produced a synergistic effect on Penh. By 24 hours after the last OVA or PBS challenge, any enhanced response induced by pollutant exposure had declined to control levels. The pollutant exposures did not significantly increase airway inflammation (assessed by bronchoalveolar lavage [BAL] fluid analysis beginning 24 or 48 hours after the final OVA or PBS challenge). Because CAPs are a heterogeneous mixture of particles, elemental analysis was conducted and associations between specific elemental groupings (present in daily samples) and airway responsiveness were analyzed. This analysis showed that increased Penh in asthmatic mice exposed to CAPs plus O3 was associated with the AlSi fraction of CAPs. No such association was found in control mice or in asthmatic mice not exposed to O3. We conclude that CAPs exposure causes an immediate, short-lived (< 24-hour), small increase in airway responsiveness in mice and that changes in airway physiology are correlated with specific elements found within the particle mixture. PMID- 16220690 TI - Improved K-means clustering algorithm for exploring local protein sequence motifs representing common structural property. AB - Information about local protein sequence motifs is very important to the analysis of biologically significant conserved regions of protein sequences. These conserved regions can potentially determine the diverse conformation and activities of proteins. In this work, recurring sequence motifs of proteins are explored with an improved K-means clustering algorithm on a new dataset. The structural similarity of these recurring sequence clusters to produce sequence motifs is studied in order to evaluate the relationship between sequence motifs and their structures. To the best of our knowledge, the dataset used by our research is the most updated dataset among similar studies for sequence motifs. A new greedy initialization method for the K-means algorithm is proposed to improve traditional K-means clustering techniques. The new initialization method tries to choose suitable initial points, which are well separated and have the potential to form high-quality clusters. Our experiments indicate that the improved K-means algorithm satisfactorily increases the percentage of sequence segments belonging to clusters with high structural similarity. Careful comparison of sequence motifs obtained by the improved and traditional algorithms also suggests that the improved K-means clustering algorithm may discover some relatively weak and subtle sequence motifs, which are undetectable by the traditional K-means algorithms. Many biochemical tests reported in the literature show that these sequence motifs are biologically meaningful. Experimental results also indicate that the improved K-means algorithm generates more detailed sequence motifs representing common structures than previous research. Furthermore, these motifs are universally conserved sequence patterns across protein families, overcoming some weak points of other popular sequence motifs. The satisfactory result of the experiment suggests that this new K-means algorithm may be applied to other areas of bioinformatics research in order to explore the underlying relationships between data samples more effectively. PMID- 16220692 TI - [Introduction to myocardial doppler imaging]. AB - Introduced in 1989, the myocardial doppler tissue imaging has now many clinical applications. Doppler tissue imaging explores the proper systolic and diastolic regional function of myocardial fibers. We can measure myocardial velocities by recording signals of low velocitiy and high magnitude, the strain and the strain rate are then derived from velocities. Specific software is therefore necessary. There are many ways of recording and representing myocardial doppler tissue parameters. Diagnostic and prognostic value of myocardial doppler tissue imaging is now proved and this method has many applications in ischemic cardiopathy and in cardiomyopathies. The development of powerful software is promising for the applications of the strain in the future. PMID- 16220693 TI - [Isoniazid acetylation in a group of Tunisian patients. Report of 620 patients]. AB - Isoniazid is a first line antituberculosis drug metabolized mainly in the liver by the Nacetyltransferase. There are differences between individuals in acetylation metabolism. Subjects are thereby characterized as being rapid or slow acetylators. The purpose is to study the distribution pattern of acetylation in patients with tuberculosis followed up at the teaching Hospital of La Rabta. The determination of acetylator phenotype was carried out on 620 tuberculosis patients during a period of 12 years. There were 483 men and 137 women with a median age of 40.3 years. The test was investigated before drug regimen administration at the dose of 5 mg/kg. A blood sample was taken three hours after the first administration. The determination of acetylation profile was worked out by Vivien hypothesis. In our population 391 were low and 229 were fast acetylators. The median dose recommended within the test was 3.04 mg/kg/day. 56% of our patients were initially receiving high dose of isoniazid. An increase in serum transaminase was initially observed in 60 patients among whom 47 slow acetylators. After dose adaptation, 53 patients had improved their biological abnormalities. The majority of Tunisian population seem to belong to slow acetylators modal. The frequency of hepatotoxicity suggests reducing the recommended dose of isoniazid from 5 to 3 mg/kg/day. PMID- 16220694 TI - [Disposal of insulin syringes by diabetic patients. Report of 100 patients]. AB - Elimination of syringes and needles is regulated in hospitals. However, the regulations remain unknown of diabetic patients who do not eliminate correctly their syringes of insulin. We propose to analyze the present situation concerning the management of used syringes of insulin by diabetic patients. This study was conducted in a clinic for diabetics and it involved 100 diabetic patients treated with insulin. The results of this study shaved an insufficiency in the management of syringes of insulin by diabetic patients. Thus, education and a conscionsness raising of diabetic patients on management and correct use of syringes are necessary. PMID- 16220696 TI - [Do prothrombin time, transaminases, and platelet count predict the severity of fibrosis in viral liver diseases?]. AB - The goal of our study is to assess the performance of a score combining 3 serological parameters (PT, platelets and transminases) in predictive hepatic fibrosis. The study involved 54 patients, 29 of which resulted in distributing them according to the score of Metavir into 2 groupes: Group A with mild fibrosis (F1,F2) and group B with severe fibrosis (F3,F4). PT, platelets and transminase values in serum were assessed for all patients. The total score of the 3 biological parameters combined, was significantly more elabled in the severe fibrosis group than in the mild fibrosis group (P = 0.01) but it was not of much importance in 30% of cases as it did not allow to classify the patients with certainty in group A or in group B. This score does not seem to grant exemption from needle liver biopsy, but it can be improved by the association of other direct markers of fibrosis. PMID- 16220695 TI - [Current concepts in the management of pyogenic brain abscesses]. AB - Brain abscesses are severe infections with lethal outcome in the case of delay in diagnosis and treatment. The authors report their experience about 30 patients with pyogenic brain abscesses treated between 1989 and 2000. The goal of the study is to analyse epidemiological, clinical and therapeutic aspects of this disease. The sex ratio (M:W) was 2,3 and the mean age was 34 years. Predisposing factors were sepsis of adjacent organs (53%) and bacteremia (30%). The clinical manifestations were: fever (83%), headaches (90%) and focal neurologic signs (57%). CT-scan findings were single lesions in 80% of cases. CT-scan showed a deep ring-enhancing lesion with surrounding edema in 77% of cases. Bacteriological agents in 13 cases (43%) were: 4 Staphylococcus aureus, 2 Neisseria meningitidis and 7 negative Gram bacilli. Medical treatment alone was in 77% of cases; seven patients benefited from medical and neurosurgical approaches. Death occured in 10% of cases. PMID- 16220697 TI - [Tracking of cardiovascular risk factors among school children: a four-year population surveillance in Susa, Tunisia]. AB - In order to confirm the phenomenon of tracking of cardiovascular risk factors among school children, we undertook in 2003, a prospective survey of a population of 789 pupils aged 13 to 15 years who had participated in a first investigation on cardiovascular risk factors in 1999. We were able to follow and study 453 pupils (57.4% of the initial population). Prevalences of hypertension and obesity were respectively 11.3% and 6.1% without significant difference between sexes. On the other hand, hypercholesterolemia was significantly more elevated among girls than boys (16.1% versus 9.3%, p = 0.039). Children classified initially as hypertensive, obese or having a hypercholesterolemia were identified 4 years later at a subsequent exam as hypertensive, obese or having a hypercholesterolemia in respectively 25%, 48.9% and 59%. The stability of cardiovascular risk factors among children imposes an early screening for a better management and a policy of cardiovascular health promotion since childhood based essentially on prevention of risk factors in order to reduce the cardiovascular disease burden in adulthood. PMID- 16220698 TI - [Osteopetrosis with carbonic anhydrase II deficiency: report of 24 cases]. AB - Twenty four patients suffering from osteopetrosis caused by carbonic anhydrase II deficiency are colliged. This pathology seems to be frequent in Tunisia. Mental retardation is present in 52%, 85% of patients have short stature and 25% have optic atrophy. All affected subjects show craniofacial disproportion and dental anomalies. Twenty patients have at least one bone fracture. Metabolic acidosis is constant: it is profound during the first life decade. A severe selective reduction of carbonic anhydrase II in erythrocyte is confirmed in 18 cases. Osteosclerosis and defective skeletal modelling are constant, cerebral calcification can be seen at the scanner approximately at the age of two years and six months. All patients are homozygous for a splice junction mutation in intron 2 of the carbonic anhydrase II gene, this mutation does not seem to protect patients from bone fractures nor induce a severe metabolic acidosis. PMID- 16220699 TI - [Allergy to food colouring. A prospective study in ten children]. AB - In order to know more about allergy to food colouring, we conducted a prospective open study over 8 months in a group of 10 atopic children with repeated urticaria. The mean age was 6.5 years (4.2 - 13.5 years). The diagnosis was based on oral challenge tests along with hisory taking there were 3 cases of allergy to food colourings. Clinical features were mainly skin symptoms, sometimes associated with GI manifestations which were not only rare (1 case in our series) but also non specific. Colourings-free diet was recommended in consequence. It resulted in the disappearance of the symptoms in a child (Red cochineal) and their regression in the two others (Red cochineal, Red beet) with a follow up of 8 months and 3 months respectively. PMID- 16220700 TI - [A particular hepatocellular carcinoma combining the ordinary and the fibrolamellar variant]. AB - Fibrolamellar carcinoma (FLC) of the liver is a rare variant of hepatocellular carcinoma (HCC) occurring on non cirrhotic liver. Since its first description by Hugh Edmondson in 1956, 200 cases of FLC have been reported in the literature, but only some cases describe the association of the ordinary HCC with the FLC within the same lesion. We report in this study the case of a 14-year-old female patient with a hepatic mass whose radiological aspect evoked a nodular and focal hyperplasia. Histologically, this tumor was composed of area of FLC mixed with ordinary HCC. Staining for cytokeratine 7 was positive in the FL component and negative in the ordinary HCC component. PMID- 16220701 TI - [Anorectal malignant melanoma. Report of three cases]. AB - The authors report three cases of primary anorectal malignant melanoma in order to discuss the various diagnostic problems, therapeutic modalities and to remind of the prognostic factors of this rare and unknown affliction. The diagnosis is unfortunately realized in the advanced stage. Mrs B.O, 55 years old, presented rectal hemorrhages and false meeds since a year, the clinical examination showed rectal tumor that bleeds with touch. The mass has been biopsed during the rectoscopy and the diagnosis of the malignant melamoma has been confirmed. Abdominoperitoneal amputation had been realized. Mr F.K, 35 years old, hospitalized because of constipation and rectal hemorrhages that evolve since 7 months with loss of weight and alteration of the general state. The rectal touch emphasizes a budy rectal polypoid tumor about 6 cm that the biopsies confirmed the diagnosis of invasive malignant. A Hartman's operation has been realized. A resection of the tumoral bud has been realized 3 months later, the patient died 4 months after that. Mrs F.K, 50 years old, presented since 50 days relapsing rectorrhages. The rectal touch showed a rectal tumor far about 6 cm from the amal margin, the biopsy during the rectoscopy confirmed the diagnosis of the pigmented and little invasive malignant melanoma. The abdominal exhography showed hepatic metastases and a resection by endo-mal way in order to reduce the tumor has been realized. The inclusion of the primary anorectal malignant melanoma in the diagnosis of the afflictins of the anorectal region would permit an improvement of this affliction prognosis, this is still unfortunate when the diagnosis is late. Its treatment is still surgical, the role of the other therapies still needs to be defined. PMID- 16220702 TI - [Intradiploic hematoma: report of two cases and review of literature]. AB - Diploic hematoma is an uncommon lesion which develops generally after minor head trauma. Only seven cases have been published so far and we are reporting two new cases of diploic hematoma in infancy. The diagnosis was made by computerized tomography and confirmed by histology. Diploic hematoma should be considered as part of the differential diagnosis in case of with a diploic lesion. PMID- 16220703 TI - [Childhood linear psoriasis]. AB - Linear distribution of psoriasis is rare. This presentation offers to physicians some diagnostic difficulty, especially in the absence of a history of pre existing psoriasis or in the presence of any other linear dermatosis. In this study, we report cases of 3 girls, ages 4 yr, 5 yr and 10 yr, admitted to our dermatology department. The clinical features and differential diagnosis of this skin disease which, in children, can be easily mistaken for inflammatory verrucous epidermal nevus, are discussed. PMID- 16220704 TI - [Secondary plasma cell leukemia. Report of 2 cases]. AB - Plasma cell leukemia is considered as the leukemic variant of multiple myeloma. It is a rare entity. There are two forms: a secondary one following a known myeloma, the diagnosis of which is easy, and a primary one arising without a preceding phase of multiple myeloma. The diagnosis of the latter form is more difficult, a differential diagnosis has often to be discussed with other lymphoproliferative diseases. Prognosis is poor. We report 2 cases of secondary plasma cell leukemia diagnosed over ten years, among 59 of multiple myeloma cases. We describe the epidemiologic, clinical, biological and evolutionary characteristics. PMID- 16220705 TI - [Pemphigoid gestationis: a pregnancy-related pathology underestimated by obstetricians]. AB - Pemphigoid gestationis (herpes gestationis) is an autoimmune bullous dermotosis occuring dining pregnancy or in the post-partum period. The interaction of this rare pathology with pregnancy is underestimated by obstetricians. Authors report one case of pemphigoid gestationis diagnosed in a pregnant woman at term. After a literature review, clinical aspects of this dermatosis and especially its interactions with pregnancy are assessed. PMID- 16220706 TI - Lead, follow or get out of the way. PMID- 16220707 TI - Spa dentistry: all this and dentistry, too. PMID- 16220708 TI - Drug-induced osteonecrosis of the jaws: a new clinical entity. PMID- 16220709 TI - How to make money even when you're out of the office. PMID- 16220710 TI - Taking care of business. PMID- 16220712 TI - Vietnam 30 years later. PMID- 16220711 TI - Duke study reports sex, self esteem diminish for morbidly obese. PMID- 16220713 TI - The evolution of the oral hygiene. PMID- 16220714 TI - Which snacks can damage your teeth? PMID- 16220715 TI - The difference makes the difference. PMID- 16220716 TI - Evolution: advancing the environmental health professional step by step. PMID- 16220717 TI - Prevalence of elevated blood lead levels and evaluation of a lead-risk-screening questionnaire in rural Minnesota. AB - The objectives of the study reported here were to determine the prevalence of elevated blood lead levels in rural Minnesota and to evaluate a lead-risk screening questionnaire. Blood lead tests and lead risk questionnaires were obtained for a sample of 1,090 children younger than 48 months of age (72 percent of the eligible population) from three rural counties of west-central Minnesota between September 1, 2001, and August 31, 2002. It was found that overall, 2.4 percent of children in the study had blood lead levels of > or = 10 microg/dL (0.48 micromol/L) (results for capillary and venous tests combined), 0.9 percent had venous blood lead levels of > or = 10 microg/dL, and 0.5 percent of study participants had blood lead levels of > or = 20 microg/dL (0.96 micromol/L). Three risk factor questions, when taken together, predicted 90 percent of blood lead levels of > or = 10 microg/dL and all blood lead levels of > or = 20 microg/dL. The study estimated the prevalence of lead poisoning using a sample of the entire population rather than a clinic-based convenience sample. The authors conclude that targeted screening is an effective way to identify lead-poisoned children in rural areas of Minnesota. PMID- 16220718 TI - The influence of well construction on bacterial contamination of private water wells in Pennsylvania. AB - Pennsylvania has a large population that relies on private water wells, but no statewide regulations govern the location or construction of the wells. Bacterial contamination occurs in nearly half of these water supplies, representing a health risk to rural residents. The role of large-scale aquifer contamination versus wellhead contamination from poor construction is poorly understood. The authors disinfected and installed sanitary well caps on 16 wells that contained coliform bacteria and retested them after 30-60 days and one year. Coliform bacteria were present in seven of the 16 wells within 30-60 days and all but two wells within one year. The occurrence and prevalence of bacteria in wells appeared to be influenced by weather conditions during the study. The few wells where disinfection and installation of a sanitary well cap were successful had low initial coliform bacteria and no E. coli. Samples were also collected from 24 new wells that were constructed with a grout seal and sanitary well cap. Twenty nine percent contained coliform bacteria, and 17 percent contained E. coli. The results from this limited number of wells suggest that bacterial contamination of private wells in Pennsylvania may often originate from areas away from the wellhead. PMID- 16220719 TI - Reducing bioaerosol dispersion from wastewater treatment and its land application: a review and analysis. AB - Wastewater treatment systems and spray irrigation of treated water may spread microorganisms such as bacteria and viruses through dispersion of aerosol particles. A recent review (Brooks, Josephson, Gerba, & Pepper, 2004) identifies appropriate reports. Teltsch and co-authors report findings that suggest effective management controls involve providing buffer zones, irrigating in the daytime and in times of low humidity, reducing microorganism levels in water used for spraying, and testing for multiple types of viruses and bacteria (Teltsch & Katzenelson, 1978; Teltsch, Shuval, & Tadmor, 1980; Teltsch, Kedmi, Bonnet, Borenzstajn-Rotem, & Katzenelson, 1980). Camann, Moore, Harding, and Sorber support these findings. They also note that fecal streptococci are hardier than fecal coliform and appear frequently in background samples, suggesting that this bacterium is a better indicator of background and downwind conditions than are fecal coliform bacteria. In their study, storage prior to spray irrigation reduced microorganism concentrations by 99 percent. Downwind concentrations of sprayed reservoir water were often comparable to background values (Camann, Moore, Harding, & Sorber, 1988). Italian researchers (Brandi, Sisti, & Amagliani, 2000; Carducci, Gemelli, Cantiani, Casini, & Rovini, 1999; Carducci et al., 2000) confirm variable die-away rates of microorganisms, observe a positive association between fecal streptococci and the presence of viruses, and recommend consideration of submerged aeration for sludge digestion at sewage treatment plants. No reports are available that measure dispersion of bioaerosols from wastewater consistently treated to meet contemporary disinfection standards. PMID- 16220720 TI - Nitrogen dioxide increases cardiorespiratory admissions in Torrelavega (Spain). AB - The objective of the study reported here was to analyze relationships between levels of air pollutants and emergency admissions for cardiorespiratory disease. Admission data from January 1, 1992, to December 31, 1995, were obtained from the Marques de Valdecilla University Hospital Admission Service; meteorological data (rainfall, temperatures wind speed, wind direction) were obtained from the National Meteorology Institute in Santander. Pollutant data on sulfur dioxide (SO2), hydrogen sulfide (H2S), total suspended particles (TSP), nitrogen oxide (NO), and nitrogen dioxide (NO2) were provided by the secretary of environment for the Cantabrian Regional Government. Rate ratios were estimated for each pollutant by Poisson regression; they were adjusted for meteorological variables. It was found that elevated NO2 increased by 20 percent the risk of having an admission for cardiorespiratory diseases; this effect was mainly due to respiratory diseases (rate ratio = 1.7, p < .001) and was negligible for cardiac diseases (rate ratio = 1.1, p = .28). In the one-pollutant model, elevated particulates and nitrogen monoxide were also related to admissions, but this effect disappeared when a five-pollutant model was used (p = .21 and p = 0.36, respectively. SO2 and SH2 did not show any relationship with admissions. Thus, nitrogen dioxide was the only pollutant the authors found to be related to emergency admissions for cardiorespiratory diseases. It is difficult to generalize from these results because of the small number of daily admissions and the variability in pollutant levels; therefore, more studies are necessary to improve knowledge about the relationship between air pollution and health in small towns. PMID- 16220721 TI - An introduction to liability, negligence, and all things in between: Part I. PMID- 16220722 TI - Mean street. PMID- 16220723 TI - Pregnancy can be a time of joy, but also of potential conflicts. PMID- 16220725 TI - Later and lower. PMID- 16220726 TI - Complementary medicine. PMID- 16220727 TI - What works in breastfeeding promotion? PMID- 16220728 TI - Is breast best? Is early solid feeding harmful? PMID- 16220729 TI - Promotion of periconceptional folic acid has had limited success. PMID- 16220730 TI - Pre-eclampsia: how to find the needle in the haystack. PMID- 16220731 TI - Nicotine replacement therapy in pregnancy: use or avoid? PMID- 16220732 TI - Effects of substance abuse during pregnancy. PMID- 16220733 TI - Keeping the needs of teenage parents at the heart of services. PMID- 16220734 TI - Stress and pregnancy. PMID- 16220735 TI - Health promotion in pregnancy: the role of the midwife. AB - Health promotion is of particular importance to midwives who promote health rather than manage disease and ill health. Although the midwife has always had a role in public health, there is now an explicit need for the profession to direct its attention to teenage pregnancy, smoking cessation, drug awareness and domestic violence. Much of the role of the midwife during pregnancy is in health promotion and a more explicit application of such may carry benefits in meeting Government policy on public health. Some activities undertaken by midwives may not be identified as health promotion, though there is evidence that the interaction generated by routine examinations is of benefit to the mother's health. Midwives should work in partnership with women and families, facilitating decisions about the care that they feel they may require. Social disadvantage may impede participation where formal education was not valued or ethnic background or language impaired access to traditional childbirth education. Tackling this is at the heart of current public health policy around childbirth and child care. Education can take place during any interaction and this gives midwives huge scope to provide an educational experience for women each time they meet. For the pregnant teenager the extended family may need to be included in health promotion activities particularly if breastfeeding targets are to be met. A united health and education policy to inform and educate children and teenagers about the benefits of pre-conceptional care and breastfeeding may be needed. In this way young women come into contact with midwives before they are pregnant, before attitudes to breastfeeding are established and before the concept of pre conceptional care is lost. Although breastfeeding improves health for women and their infants it can become another burden and expectation which they fail to achieve. Professionals need to be sensitive to the possible negative impact on a woman's health, which could be reduced if the emphasis was moved from individual behaviour change to the inequalities within society. Midwives should seek to respond positively to service changes to achieve the goal of multidisciplinary, non-hierarchical patient-centred services. In facilitating change midwives seek to use their influence to the benefit of the pregnant woman. PMID- 16220736 TI - Prevention of postnatal depression. AB - Postnatal depression is the most frequent psychiatric disorder seen after childbirth, with a prevalence rate of 10% to 15%. The women at risk need to be identified by a valid and reliable method, either using a screening instrument or an interview schedule. The preventive strategies need to have enough power to detect a clinically worthwhile effect to be considered useful in clinical practice. Many of the risk factors for developing postnatal depression are present during the pregnancy and immediate post-partum period. The risk factors for postnatal depression include depression or anxiety during pregnancy, experiencing stressful life events during pregnancy or the early puerperium, maternity blues, low levels of social support, past history of depression and poor marital adjustment. The antenatal and postnatal period provides an ideal opportunity to screen women for these risk factors. The women identified to be at risk can be identified, and preventive interventions can be implemented. Routine clinical practice can be improved to identify some of the women at risk by better communication between health professionals. There are no antenatal screening tools that have been shown to be of benefit in predicting postnatal depression. Edinburgh Postnatal Depression Scale is widely used in the postnatal period to screen for depression. The psychosocial interventions to prevent postnatal depression have not been shown to be beneficial and there is a dearth of psychopharmacological trials to make firm conclusions about their efficacy in preventing postnatal depression. Individualised psychosocial interventions aimed at the at-risk populations and initiated in the postnatal period appear to have some benefit in preventing postnatal depression. The focus of this article will be the risk factors associated with postnatal depression, screening methods and tools to identify those at risk of developing the disorder and the psychosocial, psychological and psychopharmacological interventions to prevent postnatal depression. PMID- 16220737 TI - Factors influencing antenatal and postnatal diets of primigravid women. AB - This article disseminates research on the self-reported perceived factors influencing antenatal and postnatal diets of a sample of primigravid women. Participants (n = 39) were recruited from general medical practices during the first trimester of pregnancy, with 37 participants remaining in the study until six months post-partum. Self-reported diet diaries at the first trimester, third trimester and six months post-partum generated information on food consumption, using estimated portions. In-depth interviews at the same three time points identified environmental, physical, social and emotional barriers to healthy eating during pregnancy. Findings from the interviews indicate that the key influences on dietary choices change over the three time periods. Conclusions relate to the key influencing factors and a number of recommendations are made for health promotion interventions to ensure women are informed and able to choose a healthy eating plan during pregnancy and post-partum. PMID- 16220738 TI - The nutritional status of women in the first trimester of pregnancy attending an inner-city antenatal department in the UK. AB - We have previously found high rates of poor iron and folate status in women who had delivered a low birthweight baby (LBW) in an ethnically diverse inner-city area of the UK. However, little was known of the nutritional status in the local general obstetric population. We therefore investigated biochemical measures of nutritional status in the first trimester of the first pregnancy. Routine blood samples collected at the antenatal booking clinic were analysed for haemoglobin (Hb), serum ferritin, red cell folate (RCF) (n = 100) and erythrocyte transketolase activation coefficient (ETKAC) for thiamin status (n = 90). We found 9% of women in our sample had a low Hb level, 10% had a low serum ferritin and only one had a low RCF. This is a substantially lower number of women with biochemical deficiencies than we found previously in women three months after delivering a LBW baby. However, 34% had low thiamin status. Thiamin status was negatively correlated with gestational age at birth (r = -0.407, p < 0.001). Differences in nutritional status were observed between ethnic and socio-economic groups. Hb levels differed between ethnic (p = 0.001) and socio-economic groups (p = 0.02), with Africans and women in manual occupations/unwaged having the lowest Hb levels. RCF levels also differed between groups (p < 0.001) with Caucasians and those in non-manual occupations having highest levels. ETKAC also differed between ethnic groups (p = 0.008) with Africans having the highest level indicating a poorer status. The study highlights the need to improve nutrition particularly in ethnic minorities and low income groups who are most at risk of adverse birth outcomes such as LBW. PMID- 16220740 TI - [Hypertension: the year 2004]. PMID- 16220739 TI - Trends in cesarean rates for first births and repeat cesarean rates for low-risk women: United States, 1990-2003. AB - OBJECTIVES: This report presents trends in cesarean rates for first births and repeat cesarean rates for low-risk women, in relation to the Healthy People 2010 (HP 2010) objectives. Data for the U.S. showing trends by maternal age and race and Hispanic origin are presented. METHODS: Cesarean rates were computed based on the information reported on birth certificates. RESULTS: With a decrease between 1990 and 1996 and an increase between 1996 and 2003, the trend in the cesarean rate for low-risk women having a first birth paralleled trends in the primary (regardless of parity) and total cesarean rates. During 1996-2003 the cesarean rate for low-risk women having a first birth has consistently been at least 13 percent lower than the rate for all women having a first birth. For 2003 the cesarean rate for all primiparous women was 27.1 percent; for low-risk women the rate was 23.6 percent. The trend in the repeat cesarean rate for low-risk women was similar to the trend in the repeat rate for all women, i.e., a decrease from 1990 to 1996 and an increase from 1996 to 2003. The repeat cesarean rate for low risk women has consistently been slightly lower than the rate for all women. For 2003 the repeat rate for all women was 89.4; the rate for low-risk women was 88.7. These trends were found for low-risk women of all ages and racial or ethnic groups. Therefore, low-risk women giving birth for the first time who have a cesarean delivery are more likely to have a subsequent cesarean delivery. PMID- 16220741 TI - [Prognostic value of QKD interval corrected by QRS duration in hypertensive patients]. AB - OBJECTIVES: The QKD interval is measured between the onset of QRS on the ECG and detection of last Korotkoff sound by a microphone placed on the brachial artery while measuring BP. It is the sum of preejection time and pulse transmission time and thus is correlated to pulse wave velocity (PWV). This interval is automatically monitored with BP and HR every 15 minutes during 24 h with an ambulatory blood pressure monitoring device (Diasys integra, Novacor, France). The 96 measurements obtained allow to automatically calculate the QKD(100-60), QKD value for 100 mmHg SBP and 60 bpm HR. This indice of arterial stiffness has been shown to be linked to future cardiovascular (CV) events, independently of 24 h BP. However this interval may be abnormally prolonged in case of left bundle branch blocks (LBBB). METHODS: We tested the effects of simply removing QRS duration from QKD(100-60) value on the prediction of CV events in a population of 412 hypertensives (247 males: age = 53 +/- 14 years; office BP = 158 +/- 19/97 +/ 11 mmHg; 24 h BP = 133 +/- 17/86 +/- 11 mmHg) followed prospectively. RESULTS: Mean follow up was 65 months, 32 pts were lost, 49 CV events occurred including 11 deaths. Cox model showed that baseline QKD(100-60) (m = 202 +/- 19; 142-254 ms) was significantly (p < 0.05) associated to events independently of age, 24h SBP and other traditional risk factors. Removing QRS duration (m = 85 +/- 10: 61 158 ms) improves the relation to events (monovariate khi2 = 38 vs 30). CONCLUSION: Removing QRS duration from QKD(100-60) improves its predictive value of future CV events and allows using this method in patients with LBBB. PMID- 16220742 TI - [Is masked hypertension an artefact due to the blood pressure measurement method and threshold effects?]. AB - From results of office and home measurements of blood pressure (BP), patients can be classified as "hypertensive (HT)", "normotensive (NT)", "office hypertensive (OH)" or "masked hypertensive (MH)" by crossing the classifications obtained from each method. It seems that 9 to 20% of patients could be MH with a prognosis close to HT (SHEAF study). OBJECTIVES: To test the hypothesis that at least one part of the prevalence of MH would be an artefact due to the difference between the methods of measurements (shygmomanometer vs semi-automatic device) and/or due to different definitions of office hypertension (OHT). To determine the impact of different definitions of OHT on the prevalence of MH. METHODS: During the course of a phase IV study, BP was measured with the same semi-automatic device (OMRON 705CP) both at doctor's office (3 measurements at 1-minute intervals) and at home, by the patient himself (3 measurements in the morning and in the evening at 1-minute intervals over the 7 days before the visit). Following definitions were used: Office HT: SBP > or =140 mmHg, DBP > or =90 mmHg, SBP > or =140 mmHg or DBP > or =90 mmHg; Home HT: SBP > or =135 mmHg, DBP> or =85 mmHg, SBP > or =135 mmHg or DBP > or =85 mmHg. Another definition of office HT was used SBP > or =135 mmHg, DBP > or =85 mmHg SBP > or =135 mmHg or DBP > or =85 mmHg. RESULTS: 575 patients were analysed. Results from the two methods of measurements are closed but significantly different (difference for SBP: 3.2 +/- 16.5 mmHg; p < 0.0001; difference for DBP: 1.4 +/- 10.3 mmHg; p = 0.002) PMID- 16220743 TI - [Aortic arch shape and flow dynamics]. AB - Post-operative deformation of the aortic arch architecture is associated with an increased risk of hypertension following correction of coarctation. In addition to morphological analysis, MRI allows a functional analysis of the thoracic aorta. We report three examples which illustrate the direct relationship between aortic arch morphology and blood flow in the thoracic aorta. PMID- 16220744 TI - [District program to improve the cardiovascular risk of resistant hypertensive patients in general medicine]. AB - PURPOSE: To try to improve the cardiovascular risk of resistant hypertensive patients in general medicine in Brittany after using french hypertension recommendations. METHODS: 581 hypertensive patients under 3 antihypertensive drugs have asked for an exoneration of the patients' social contribution: 297 (51%) were uncontrolled at their general practitioner (GP) among whom 106 (36%) have refused to take part in the program. 191 resistant hypertensive patients followed by 170 different GP have been pre-included. After a meeting between the social security physician, the GP measured blood pressure (BP) with a validated BP device and passed it on to the patient for a self-BP measurement (SBPM). In the case of a confirmed resistance, after specialist opinion, the treatment was then modified and the patient was checked after four months by his GP for a new clinical and self BP measurement. RESULTS: After the initial automated BP measurement by GP, 51 patients (27%) had in fact controlled hypertension. Out of 136 resistant hypertensive patients (mean: 63 ys) during consultation (60% with systolic isolated hypertension), 121 (89%) have been closely followed during the whole study period among who 114 were also resistant with SBPM (94%). Only 8 patients (6%) were controlled at home. A check-up with specialist opinion was conducted: ECG (82%), echocardiography (59%), vascular echography (35%), funduscopy (30%), plasma renin/aldosterone measurement (15%), renal artery exploration (26%). Eight (7%) secondary hypertension have been founded. After 4 months, 32 (26%) obtained controlled hypertension during GP consultation and 15% during SBPM but the 20/32 controlled patients (62%) had a masked hypertension. Cholesterol levels (63% of dyslipidemics) and the body mass index (80% of overweighed patients) have not varied. On the contrary, 12/28 (48%) has stopped smoking. The coronary risk using Anderson's model has only decreased from 16.5% to 13.8%. CONCLUSION: If this health program has shown its feasibility and the good participation of the GPs, treatment of hypertension and others risk factors really remains insufficient. On the contrary, the use of a validated automatic BP device has been really well carried out. Its use has allowed reclassifying as controlled one third of the resistant hypertensive patients and as uncontrolled at home one patient out of two which were controlled with GP. These results prove the necessity to increase this BP measurement technique in this population. PMID- 16220745 TI - [Relationship between aortic arch shape and blood pressure response after coarctation repair]. AB - The mechanisms of secondary hypertension after repair of coarctation of the aorta are not well understood. Abnormalities of the architecture of the aortic arch and their consequences on blood pressure have not been studied. In order to study the relationship between abnormalities or aortic arch architecture and resting blood pressure ninety-four patients without re-coarctation were followed up prospectively from 1997 to 2004 (mean age 16.9 +/- 8.1 years; mean weight 57.5 +/ 18.3 Kg; interval since surgery 16.3 +/- 5.4 years). All underwent MRI angiography of the thoracic aorta which enabled the abnormalities to be classified in 3 groups: gothic arch, crenellated arch and roman arch. Twenty-four patients (25.5%) were hypertensive and 70 (74.4%) normotensive. There were 40 gothic arches (42.5%). 14 crenellated arches (15%) and 40 roman arches (42.5%). Gothic arches were more commonly observed in the hypertensive patients (18/40, [45%, 95% CI 31-62]) than the crenellated arches (4/14, [28.5%, 95% CI 7-48]) or the roman arches (2/40, [5%, 95% CI 2-12]). Only the gothic arch was independently correlated with hypertension on multivariate analysis. The authors conclude that gothic deformation of the aortic arch is an independent predictive factor of hypertension in patients operated for coarctation with an excellent result on the isthmic region. Patients with a gothic appearance of their aortic arch should be followed up closely. PMID- 16220746 TI - [Systolic blood pressures are not comparable when home blood pressure is measured with a wrist or an arm validated monitor]. AB - OBJECTIVES: To compare home blood pressure values obtained with two validated OMRON (wrist or arm) monitors used sequentially in the same subject. METHODS: In 265 hypertensive subjects referred to hypertension specialists, a self measurement of blood pressure was performed sequentially with an OMRON M4-I (arm cuff, A/A, BHS validation) or OMRON RX-I (wrist cuff, B/B, BHS validation). Each patient recorded home blood pressure during two periods of 4 days with 3 measures in the morning and 3 in the evening. Order for use of each monitor was randomised. With wrist devices, subjects were advised to keep the arm at heart level during measurements. BP values were reported on a standardized document. Patients were asked by a questionnaire about the tolerance and feasibility of the 2 methods. RESULTS: In this population, aged 59 +/- 14 years, with 60% of men and a mean blood pressure of 152 +/- 21 / 86 +/- 14 mmHg, the home blood pressure values were 143 +/- 20/81 +/- 11 mmHg with the arm monitor and 135 +/- 10 / 80 +/ 11 mmHg with the wrist monitor. Mean SBP adjusted on age, initial blood pressure level and period order was significantly lower when home blood pressure monitoring has been recorded with a wrist monitor as compared to an arm monitor (p < 0.001). Self measurement of blood pressure was felt as easy in 92% with the arm monitor and in 96% with the wrist monitor (p < 0.05). Self measurement of blood pressure was felt as constraining in 14% with the arm monitor and in 7% with the wrist monitor (p < 0.01). The feasibility between the two devices was good with none of the value missing in 86% with the arm monitor and in 85% with the wrist monitor. The missing values were in 56% the fourth day. CONCLUSION: Despite the use of two validated monitors, mean SBP is significantly lower when home blood pressure monitoring is recorded with a wrist monitor as compared to an arm monitor. Uncertainty in the arm position with the use of wrist device could explain these results. When advising home blood pressure monitoring, care should be taken to recommend only the use of validated devices and to prefer the use of arm devices in order to avoid the uncertainty of an inadequate utilisation. PMID- 16220747 TI - [Use of home blood pressure devices in France in 2004]. AB - OBJECTIVE: To estimate the number of devices for home blood pressure measurements in the French population in 2004 and to evaluate the use of these apparatus. METHODS: The French League Against Hypertension Survey 2004 (FLAHS-2004) was performed cross-sectionally on 3707 subjects, part of a sample of 5476 subjects selected as being representative of the French metropolitan population for age (35 years and above), gender, socioeconomic status, and place of living. Subjects who declare to take an antihypertensive medication were classified as treated hypertensive patients. A questionnaire evaluating the condition of use of HBP device was given. RESULTS: In 2004, 24% of the French population above the age of 35 years was treated for hypertension, corresponding to an estimate of 7.5 million of subjects. 25% of treated hypertensives and 12% of untreated subjects had a tensiometer. A total of 4 millions of BP devices are owned in the general population with 43% by treated hypertensive patients. 67% of BP devices are wrist cuff. Medical doctors recommended home blood pressure monitoring in only 12% of subjects. CONCLUSION: In 2004, the number of subjects treated for hypertension is 1/4 of the general population over the age of 35 years in France. In this group of subjects, 1/4 possessed a tensiometer but in only 10% medical doctor recommended the use of home blood pressure monitoring. PMID- 16220748 TI - [MIRA study: epidemiology of microalbuminuria in type 2 diabetes patients and associated comorbidities]. AB - This is an observational survey screening for microalbuminuria in diabetes patients type 2. This survey takes place during the general practioner (GP) consultation in Rhone-Alpes area. It is preliminary survey. Five hundred and sixty three patients were included, consecutively, during the same week, by 185 GP. They were aged from 18 to 80 years and previously research for microalbuminuria was carried out at least one year ago. A questionnaire collecting data concerning age, weight, height, diabetic vintage and HbA1c, arterial pressure, renal function, associated risk factors as (lipids, tabacco, cardiovascular history personal and family, antihypertensive diabetic and lipid treatments. Microalbuminuria as well as urine creatinine was detected with a dipstick to calculate albumin-creatinine ratio. Patients aged 64.66 +/- 11.23, 55% were male; 51% had a microalbumuniria between 30 mL/L and 300 mL/L, but if one consider the albumin-creatinine ratio 59% had a microalbuminuria. In these patients, the diabetes history is longer and body mass index >30 is more frequent 35 vs 27%. HbA1c is similar in the two groups of patients, but patients with microalbuminuria had more often two treatments. Regarding hypertension, there are no difference between the two groups in term of blood pressure control and there is no correlation between blood pressure and albuminuria level. Nevertheless 68% of the patients were uncontrolled for blood pressure. Renal impairment (creatinine clearance <60 mL/min) was present in 26% of the whole group but more frequent in microalbuminuria patients 30 vs 23%. IN CONCLUSION: prevalence of microalbuminuria seems to be higher in this population compared to data previously reported and linked to the vintage of the diabetes mellitus. Comorbidities as well as risk factors were more frequent in patients with microalbuminuria. PMID- 16220749 TI - [Blood pressure control after a one year follow-up in high risk subject screened in occupational medicine]. AB - OBJECTIVE: To identify factors predicting the success or the failure of intervention on blood pressure in a population estimated at high risks. METHODS: The program "Coeur 2001" has analysed the absolute cardiovascular risk (ACVR Framingham) in 107 371 voluntary French railways employees. In the company, were considered at high risk (HR), subjects for whom risk was > or = to the 95th percentile of the distribution of the observed ACVR by age range: ACVR > or = 4.5% before 35 years, 12% between 35 and 45 years and 19% beyond 45 years, i.e. a total of 4 190 subjects. These subjects were warned about their risk and advised to choose and consult a physician. A two-year follow up was planned. Identical data (risk factors, ACVR, type of management and therapies) were collected during the first consultation with the occupational physician (T0), one year later (T1) and two years later (T2). RESULTS: Our work concerned 2376 employees at HR, consulting at T1. At T1, 54% of subjects were in the hight risk group (SHR) [48% when BP at T1 was < 140/90 mmHg and 62% when the BP was > or = 140/90 mmHg]. The mean decrease of the systolic BP (SBP) was 4 mmHg in the whole sample, 7.7 mmHg in subjects with normalised ACVR, and it remained stable in the group still at HR (-0.7 mmHg). At T0, blood pressure (BP) was > or = 140/90 mmHg in 55.8% of the patients and 38.4 at T1. This high BP was associated with higher frequency of diabetes (14 vs 7%) and overweight (BMI > or =30 kg/m2; 32.8 vs 19.7%). The percentage of treated hypertensive subjects had increased from 35 to 62% but one third of uncontrolled hypertensive subjects was treated by mono therapies at T1. CONCLUSION: To keep BP under control is a difficult task in routine medicine. At T1, despite a more aggressive treatment, 38% of subjects at high risk were still hypertensive subjects. PMID- 16220750 TI - [Blood pressure control in hypertensive patients with stable coronary heart disease]. AB - OBJECTIVE: To evaluate systolic blood pressure (SBP) control in hypertensive patients with a stable coronary heart disease (CHD) in general practice in France. METHODS: A survey was conducted in a sample of 206 general practitionners (GP) representative of the French medical population, in 2003 [LHYCORNE survey]. Each GP had to include 3 hypertensive patients, >18 years old, BP > or = 140/90 mmHg and/or treated for hypertension, and with evidence of CHD documented by myocardial infarction (MI) or angina pectoris (AP) [diagnosis previously established by a cardiologist]. Three office BP measurements were performed, the last two recorded. BP levels were considered as controlled by treatement if they were < 140/90 mmHg. RESULTS: 595 patients were included, 75% men mean age 66 years, 25% women mean age 73 years. All patients had a CHD: MI 46%, AP 54%; 533 (90%) had more than 2 cardiovascular risk factors: hyperlipidemia (411; 69%), smokers (375; 63%), diabetes (158; 27%). Mean BP was 140.7 +/- 14/80.8 +/- 9.7 mmHg; 553 (93%) of these hypertensive patients were treated, and 239 (40%) were considered as having a controlled SBP at the treshold of 140 mmHg: 47% in patients with previous MI and 38% with AP (p < 0.001). Diastolic BP (DBP) was <90 mmHg in 480 (81%) and pulse pressure was >65 mmHg in 202 (34%); 313 (53%) patients received a combination of three drugs or more; 354 (60%) had a beta blocker, 260 (44%) a calcium channel blocker, 237 (40%) an ACE inhibitor, 287 (48%) other antihypertensive drugs (246 diuretics, 41%); 502 (84%) received antiplatelet therapy, 403 (68%) statins. CONCLUSION: This survey shows that systolic BP is not at goal in 6/10 hypertensive patients with stable CHD suggesting there is a place for a more effective combination therapy according to evidence-based medicine. PMID- 16220751 TI - [A treatment with rosuvastatin induced a reduction of arterial pressure and a decrease of oxidative stress in spontaneously hypertensive rats]. AB - The aim of this study was to appreciate consequences of rosuvastatin administration on hemodynamic function, vascular oxidative stress and ischemia/reperfusion disorders in normotensive and hypertensive rats. At 10 weeks of age, spontaneously hypertensive rats (SHR, n=20) and normotensive Wistar Kyoto male rats (WKY, n=20) were divided into four groups and given, either vehicle or 10 mg/kg/day of rosuvastatin by gavage for 3 weeks. Systolic blood pressure was assessed every week. At the end of these treatments, vascular NADPH oxidase activity was evaluated by chemiluminescence (lucigenin 0.5 microM). Hearts were isolated and perfused according to the Langendorff method and were subjected to 30 min of global ischemia. Reactive oxygen species (ROS) produced during reperfusion were quantified by electron spin resonance (ESR) spectroscopy using a spin probe (CP-H, 1 mM). After one week of treatment, rosuvastatin reduced the arterial pressure in SHR rats (180.3 +/- 2.1, SHR vs 169.7 +/- 2.3 mmHg, SHR+rosuvastatin; p < 0.01), without lowering plasma cholesterol levels; these effects were not observed in WKY. NADPH activity was 25% higher in control SHR rat aortas compared to control WKY, and was reduced by rosuvastatin in SHR rats. In isolated rat hearts subjected to ischemia/reperfusion sequences, there was a deterioration in functional parameters in control SHR compared to control WKY hearts. Rosuvastatin decreased post-ischemic contracture in WKY hearts by 50% (41.5 +/- 7.5, WKY control vs 18.4 +/- 4.6 mmHg, WKY+rosuvastatin; p < 0.01) and increased left ventricular developed pressure. This beneficial effect was accompanied by a decrease in ROS detected by ESR during reperfusion (312.5 +/- 45.3, WKY control; vs 219.3 +/- 22.9 AUC/mL, WKY+rosuvastatin; p < 0.05). In conclusion, these results are in accordance with the hypothesis that oxidative stress plays a crucial role in the pathogenesis of cardiovascular diseases including hypertension, and demonstrate the beneficial effects of rosuvastatin. PMID- 16220752 TI - [Sodium ion inhibits angiogenesis of cultured rat aortic and coronary rings]. AB - High sodium intake is associated with the development of cardiac hypertrophy in man and rats independently of the rise of blood pressure. In addition, the reduced capillary density observed in striated muscle of rats fed a high salt diet suggests that angiogenesis is altered. The aim of the present experiments was to investigate the angiogenic capacities of vascular rings isolated from the aorta and the coronary artery of rats. Vascular rings (external diameter of 1 669 +/- 9 and 323 +/- 26 microm for aorta and coronary artery. respectively) are cultured in a three-dimensional collagen type I lattice and a standard medium (DMEM+HAMF12) containing 152 mM of sodium. Sodium ion associated with chloride or citrate is added to the standard medium to achieve a final concentration of sodium of 160 and 176 mM. The role of sodium-proton exchanger is evaluated through the addition of amiloride to the culture medium. Sprouts formed from vascular explants are counted every second day until days 8 to 10. Kinetics of new vessels formation and the number of sprouts were similar in aortic and coronary rings (83 +/- 5 and 95 +/- 5 sprouts, respectively). Elevation of sodium chloride concentration inhibits by 50 to 80% the neo-vessels formation in both the aorta and the coronary artery. Anti-angiogenic effect of the high sodium medium was not affected when citrate was substituted to chloride. Amiloride (3.10(-5) M) reduced the number of sprouts formed in the standard medium; however, it counteracted the anti-angiogenic effect of elevated sodium concentration. These results indicate that high extracellular concentration of sodium and not chloride anion is accompanied by a deleterious effect on angiogenic capacities of cultured aortic and coronary artery rings through modifications of trans-membrane sodium exchanges. PMID- 16220753 TI - [Changes in life style and drug therapy for treatment of hypertension in France between 2002 and 2004]. AB - OBJECTIVES: To estimate changes in life style and drug therapy for treatment of hypertension in France between 2002 and 2004. METHODS: The French League Against Hypertension Survey 2004 (FLAHS-2004) was performed cross-sectionally on 3 707 subjects, part of a sample of 5476 subjects selected as being representative of the French metropolitan population for age (35 years and above), gender, socioeconomic status, and place of living. Subjects who declare to take an antihypertensive medication were classified as treated hypertensive patients. A questionnaire evaluating changes in life style during the last year (physical activities, quality of food consumption, smoking, and alcohol habits) was auto administered. RESULTS: In 2004, 24% of the French population above the age of 35 years was treated for hypertension, corresponding to an estimate of 7.5 million of subjects. The two most frequent prescribed drugs are: fixed-dose combination drugs and beta-blockers in patients aged less than 75 years, and calcium antagonist and fixed-dose combination drugs in patients aged more than 75 years. Hypertensive subjects experienced: a weight loss of more than 3 kg in 11% vs 9%* in the general population, an increased in fruits and vegetables consumption in 41% vs 34%*, a decreased in cheese (21 vs 17%*), pork-butchery (46 vs 34%*) and alcohol consumption (23 vs 18%*) [*p < 0.01]. On the other hand, the percentage of subjects who increased their physical activities was low and not different in the two groups (7 vs 9%). CONCLUSION: Changes in life style were more often applied by the subjects treated for hypertension that by the general population. The dominating place that occupies today fixed-dose combination drugs indicates a change of the therapeutic practices. PMID- 16220754 TI - [Relationship between arterial pressure evolution and free hemoglobin distribution into vascular wall in guinea pigs]. AB - Free hemoglobin (Hb) present at high concentration in plasma--in case of hemolysis, hemoglobin based oxygen carrier (HBOC) administration or in case of subarachnoid hemorrhage--induce hypertension as result of vasoconstriction. In this context, we studied on an exchange transfusion (ET) model at 50% of hematocrit with a HBOC, the distribution of this Hb inside abdominal aortic wall in guinea pigs in relation with mean arterial pressure (MAP) evolution. MAP was monitored during 180 min after ET and rings of abdominal aorta were taken at different times, when modifications of MAP were important, and analyzed by immunohistochemistry and confocal microscopy. Gelofusine 4%, used as control, did not modify MAP while free Hb increased MAP that reached its maximum (53% of hypertension) at t=17 min after the end of ET. MAP started to decrease (45% of hypertension) at t =60 min after ET, and recovered its baseline value at t=180 min. Confocal analysis of the vessel showed that: at 17 min (when hypertension was maximal), free Hb was present in endothelial cells (EC) and in vasa vasorum; at t=60 min (when hypertension decreased) and at t =10 min (when hypertension disappeared), free Hb was detected still in EC, but inside all abdominal aorta wall too. These results suggest in the first time that free Hb could induce a hypertension by direct interaction with EC but would also be unable to maintain this hypertension in spite of its massive tissue distribution. PMID- 16220755 TI - [Gene transfers of kallikrein, bradykinin receptor B2 and a mutated B2 receptor stimulate neoangiogenesis in peripheral ischemia]. AB - In this work, we evaluated the angiogenic effect of the gene transfer of human tissue kallikrein (TK), bradykinin B2 receptor (B2R) and a mutated form (RB2m) in a rabbit peripheral model of ischaemia. We studied the effects of the transfection of each of these factors and the effects of their co-transfection. In New Zealand anesthetised rabbits we first induced an ischaemia of the left posterior leg by ligation-excision of the superficial femoral artery and its collaterals. Seven days later, we performed i.m. injections in the ischemic tight with transfection solutions containing either the control (pcDNA3 empty backbone) or the pcDNA3-TK, the pcDNA3-TK and the pcDNA3-B2R, the pcDNA3-TK and the pcDNA3 B2Rm. Twenty eight days later, the therapeutic effect was evaluated using ultrasonographic debitmetry of the common iliac artery, perfusion index (PI) = ischemic leg blood flow /non ischemic leg blood flow (%) and capillaries measurements i.e. capillary density: number of vessels/mm2 and the ratio of vessels/muscular fiber, in the adductors and gastrocnemian muscles. The PI was increased in each treated group vs control (32.61 +/- 5.2%), pcDNA3-TK: 59.72 +/- 2.33%; p = 0.001; pcDNA3-RB2: 55.25 +/- 2.29%; p = 0.008; pcDNA3-TK + pcDNA3-RB2: 84.77 +/- 3.15%; p < 0.001; pcDNA3-TK + pcDNA3-RB2m: 103.25 +/- 4.9%; p < 0.001. The capillary density and the vessel/muscular fiber ratio increased in a parallel with the hemodynamic in the ischemic adductors (pcDNA3-TK + pcDNA3-B2Rm > pcDNA3 TK + pcDNA3-RB2 > pcDNA3-TK = pcDNA3-B2R; p < 0.001). There was no angiogenic effect measurable neither in the non ischemic adductors (right) nor in the gastrocnemian muscles. In rabbit peripheral ischaemia, the cotransfection of TK and B2R increases the arterial flow in the treated leg and potentiates the neoangiogenesis. Cotransfection of the B2Rm cDNA enhanced the synergic effect of this therapeutic strategy. PMID- 16220756 TI - [Role of vascular calcium-activated potassium channels in the regulation of human peripheral conduit artery diameter]. AB - The role of an endothelium-derived hyperpolarizing factor (EDHF), acting through the opening of vascular calcium-activated potassium (K(Ca)) channels, in the regulation of the basal diameter of human peripheral conduit arteries has never been investigated in vivo. We measured in 7 healthy subjects the effect of the local infusion of an inhibitor of K(Ca) channels, tetraethylammonium chloride (TEA, 9 micromol/min, 8 min), on radial artery diameter (echotracking) and flow (Doppler). Endothelium-independent dilatation was assessed before and after TEA using sodium nitroprusside (SNP: 5, 10 and 15 nmol/min, 3 min each). TEA induced a decrease in radial artery diameter (2.65 +/- 0.09 to 2.52 +/- 0.09 mm: p < 0.05) and flow (9.4 +/- 1.2 to 7.4 +/- 1.1 ml/min; p < 0.01) without modification in the radial artery dilatation in response to SNP (NS). The decrease in radial artery diameter was still significant even when the decrease in flow was taken as covariate into analysis (p < 0.05). These results demonstrate the role of vascular K(Ca) channels in the regulation of basal peripheral conduit artery diameter and arteriolar tone in human strongly suggesting the involvement of an EDHF a these two levels. PMID- 16220757 TI - [Vascular beta-adrenergic remodeling in rat transgenic model over-expressing endothelial beta3-adrenoceptors]. AB - In rat thoracic aorta, the stimulation of endothelial beta3-adrenoceptors (beta AR) produces a vasorelaxation through activation of a NO synthase pathway and an increase in cGMP levels. In hypertension, a global decrease of the beta-AR response has been described. In spontaneously hypertensive rats (SHR), we have shown that beta3-adrenoceptor-mediated relaxation was not modified in SHR aorta at the age of 12 weeks, in spite of an upregulation of beta3-adrenoceptors. In order to determine the consequences of an over-expression of the beta3-AR, we have developed a transgenic rat over-expressing specifically in endothelial cells the human beta3-AR (Tg beta3). By real-time quantitative PCR, we have determined the expression level of the different beta-AR subtypes. We confirmed an over expression of the beta3-AR transcripts in Tg beta3 (ratio = 3.39 +/- 0.8; n=3 for Tg beta3 vs wild type [WT] animals). Surprisingly, we observed in Tg beta3 a decrease of beta1-AR transcripts (ratio = 0.76 +/- 0.03; n=3 for Tg beta3 vs WT animals) and no variation for beta2-AR transcripts (ratio = 1.95 +/- 0.60; n=3 for Tg beta3 vs WT animals). In aorta rings from WT and Tg beta3, the isoproterenol-induced relaxation was similar (WT: Emax = 82 +/- 6%, n=6; Tg beta3: Emax = 85 +/- 6, n=6). By contrast, in the presence of 10 microM nadolol, a beta1-, beta2-AR antagonist, the isoproterenol-induced response was significantly increased in Tg beta3 (WT: Emax = 68 +/- 6%, n=6: Tg beta3: Emax = 86 +/- 3; p < 0.01 vs WT). This effect was loss on denuded aortic rings. In conclusion, our study reported similar results to those obtained in hypertension in which a decrease of the beta-AR expression was associated to an elevation of the beta3-AR density. Moreover, this over-expression in our transgenic model is associated to a potential response induced by beta3-AR. Therefore, an activation of beta3-AR could supply the beta1- / beta2-AR decrease. Then, our transgenic model should be used to characterize the physiological consequences of this over expression as well as to determine the putative involvement of this receptor in the pathogenesis of hypertension. PMID- 16220758 TI - [Arterial hypertension in the French Caribbean regions: gender related differences]. AB - BACKGROUND: Arterial hypertension is a frequent disease, responsible for a significant morbidity, in the French Caribbean regions. Today, epidemiological studies on this topic remain few. OBJECTIVES: To analyse the prevalence, awareness, treatment and control of arterial hypertension in the French Caribbean regions. METHODS: Analysis of the INHAPAG cohort (Incidence of Arterial Hypertension in the Working population Antillo-Guyanaise) carried out in 2001, and including 6113 active subjects recruited in Guadeloupe, Martinique and Guyana. The procedure of diagnosis of arterial hypertension comprises two visits, with a series of three measurements of the blood pressure at each visit. RESULTS: Prevalence of hypertension is estimated to be 18.9% among women and 19.5% among men. The rates of treatment are higher among women (74.8 vs 34.4%) than among men, resulting in a much better control rate (61.3% vs 38.6% among men). An analysis of factors associated with the presence of arterial hypertension shows a noxious role of low education level among women. CONCLUSION: Our study finds marked differences between women and men in the treatment and control of arterial hypertension and underline the need for improving blood pressure management of the latter. PMID- 16220759 TI - Erb's palsy causation: iatrogenic or resulting from labor forces? AB - To determine whether there is direct evidence that Erb's palsy is caused by clinician-applied excessive lateral traction on the fetal head and neck, a literature review was performed. Both the medical literature outlining the maternal forces generated from uterine contractions and maternal pushing and limited reports on clinician-applied forces were examined. Research involving clinician-applied force measurements during actual deliveries has the potential for increasing our understanding of the mechanism of the injury. However, studies to date have involved only 29 deliveries and did not include measurement of vertical or maternally generated forces. During the past 15 years, studies have provided considerable indirect evidence that maternal propulsive forces are responsible for the injury leading to Erb's palsy. PMID- 16220760 TI - Laparoscopic uterine suspension for pain relief: a multicenter study. AB - OBJECTIVE: To assess the efficacy of laparoscopic positioning by ligament investment, fixation and truncation (UPLIFT) in alleviating pain in women with a symptomatic, retroverted uterus, the duration of pain relief, and operative or postoperative complications associated with the procedure. STUDY DESIGN: Between May 22, 1998, and November 16, 2000, 62 women with a retroverted uterus, chronic pelvic pain, moderate to severe dysmenorrhea and/or dyspareunia underwent UPLIFT on an outpatient basis. The patients for this prospective, cohort study were recruited from 5 private gynecologic practices. The patients were asked to rate their pelvic pain, dysmenorrhea and dyspareunia on a scale of 0-10, with 0 no pain and 10 the worst pain imaginable, preoperatively and again at 4 weeks and 3, 6 and 12 months postoperatively. The average follow-up time was 10.2 months (range, 0.2-14.9). RESULTS: The mean scores for pelvic pain decreased from 7.3 to 3.7 at 12 months (n = 46). Dysmenorrhea decreased from 7.8 to 4.4 (n = 39), and dyspareunia decreased from 8.0 to 3.3 (n=41). All p values were <0.0001. There were 5 minor intraoperative adverse events. The procedure did not result in any postoperative complications. CONCLUSION: UPLIFT resulted in significant, long term pain reduction in women with a symptomatic, retroverted uterus. PMID- 16220761 TI - Adhesion-prevention effects of fibrin sealants after laparoscopic myomectomy as determined by second-look laparoscopy: a prospective, randomized, controlled study. AB - OBJECTIVE: To examine the adhesion prevention effects of 2 types of fibrin sealant after laparoscopic myomectomy (LM). STUDY DESIGN: A prospective, randomized study (Canadian Task Force I) was conducted at a University-affiliated hospital. A total of 91 patients showing a minimal myoma > 5 cm, excluding pedunculated myomas, underwent LM alone: 32 patients in the control group, 29 in the fibrin gel group and 30 patients in the fibrin sheet group. After LM, postoperative adhesions were evaluated by second-look laparoscopy. The frequency of postoperative adhesions was the main outcome. RESULTS: The frequency of postoperative adhesions of the uterus was significantly lower (p < 0.05) in the fibrin gel group, with 20/32 (62.5%) in the control group, 10/29 (34.5%) in the fibrin gel group and 20/30 (67.7%) in the fibrin sheet group. Although no significant differences were found in the incidence of de novo adnexal adhesions, the lowest rate was found in the fibrin gel group, with 4/32 patients (12.5%) in the control group, 2/29 patients (6.8%) in the fibrin gel group and 5/30 patients (16.7%) in the fibrin sheet group. No bilateral adnexal adhesions were observed in the 3 groups. CONCLUSION: After LM for myomas as large as > or = 5 cm, postoperative adhesions were observed in > or = 50% of patients. The use of fibrin gel after LM is recommended. PMID- 16220762 TI - Extramembranous placement of an air-coupled vs. transducer-tipped intrauterine pressure catheter. AB - OBJECTIVE: To compare the rate of extramembranous placement between 2 types of intrauterine pressure catheter. STUDY DESIGN: Women were randomized to a transducer-tipped intrauterine pressure catheter or an air-coupled intrauterine pressure catheter from October 1998 to August 1999. Women were eligible for the study if their attending physicians decided to place an intrauterine pressure catheter. Our primary outcome variable was extramembranous placement. The intrauterine pressure catheter was left in place during cesarean delivery, and the position was documented. RESULTS: A total of 257 women entered the trial. Eight were excluded due to removal of the catheter before cesarean delivery, leaving 249 evaluable patients. Of those 249, 105 (44.3%) received cesarean delivery, with 41 in the air-coupled group and 64 in the transducer-tipped group. There were no significant differences in maternal age, gestational age, gravidity, duration of membrane rupture, birth weight, pharmaceutical drug use, cocaine use, oxytocin use or catheter removal due to poor function. The transducer-tipped catheter was placed outside the membranes significantly more frequently than was the air-coupled catheter (12.5% vs. 2.4%, p = 0.02). CONCLUSION: Transducer-tipped catheters were significantly more likely to be placed in the extramembranous space than were air-coupled catheters. PMID- 16220763 TI - Preoperative serum CA-125 levels in treating endometrial cancer. AB - OBJECTIVE: To evaluate preoperative levels of CA-125 for the prediction of advanced stages of uterine cancer. STUDY DESIGN: Retrospective chart review of 141 women with endometrial cancer who were treated by a single gynecologic oncologist at a community teaching hospital in North Carolina between November 1994 and September 2002. RESULTS: Ninety-three of 106 patients (87.7%) with surgical stage I or II endometrial cancer had normal preoperative CA-125 levels. Ten of 11 (91%) women with stage IV endometrial cancer had elevated preoperative CA-125 levels. High CA-125 levels and positive lymph vascular space invasion correlated most strongly with advanced stage (p < 0.01). Similar trends in correlation of CA-125 levels were seen with the highest grade and the deepest myometrial invasion. The sensitivity and specificity of a CA-125 cutoff level of 35 U/mL were 63% and 88%, respectively, with a positive predictive value of 61% and negative predictive value of 89%. CONCLUSION: Measurement of preoperative CA 125 is a clinically useful test in endometrial cancer patients. CA-125 appears to be a significant independent predictor of the extrauterine spread of disease and is a better predictor of disease than depth of invasion or grade. This evidence complements a growing body of literature that supports the strong relationship between CA-125 level and stage of disease. A CA-125 level should be included as part of the preoperative workup for all patients with endometrial cancer. PMID- 16220764 TI - Adnexal torsion: experience at a single university center. AB - OBJECTIVE: To review the management of adnexal torsion over an 11-year period at a single institution and to assess trends in the surgical approach, including laparoscopy vs. laparotomy, and ovarian conservation vs. removal. STUDY DESIGN: A chart review of all patients treated for adnexal torsion at the University of New Mexico from 1990 to 2001 was performed. RESULTS: Sixty-eight cases of adnexal torsion were identified. Laparoscopic management was accomplished in 22 cases (32%) and ovarian conservation in 14 cases (20.6%). There were no differences in the use of laparoscopy (OR 2.24, CI 0.65-7.93), or ovarian conservation (OR 1.24, CI 0.31-5.35) in patients treated before 1996 vs. later in the study period. CONCLUSION: Despite evidence supporting conservative management of adnexal torsion with laparoscopy and ovarian conservation, most cases at our institution are managed with open laparotomy and removal of the affected adnexa. PMID- 16220765 TI - Relationship of ovarian neoplasms and body mass index. AB - OBJECTIVE: To describe the distribution of benign and malignant ovarian neoplasms among overweight and obese women. STUDY DESIGN: A review of patients who presented with a preoperative diagnosis of a pelvic mass between 1996 and 2001 was performed; 1,096 patients were identified. Patients were stratified by body mass index into 3 groups: normal weight, overweight and obese. The pathologic findings in the 3 groups were compared. RESULTS: Complete follow-up was available on 668 patients. Overall, 248 patients were obese, 176 were overweight, and 244 had a normal body mass index. A significant difference existed in the pathologic findings in the 3 groups (p = 0.049). Women with normal body mass indices were more likely to have malignant ovarian tumors (35.2%) than were the overweight (23.9%) and obese (25.8%) women. Conversely, borderline ovarian tumors were less frequent in women with body mass indices of <25 (5.7%) than in the overweight (13.1%) and obese (10.9%) patients. Benign ovarian neoplasms occurred in 20-25% of the women. CONCLUSION: Significant differences exist in the distribution of ovarian neoplasms among women with different body mass indices. Obese women are more likely to have ovarian tumors of low malignant potential, while women with normal body mass indices more commonly have invasive ovarian tumors. Body mass index may be an important factor in preoperative counseling and risk assessment. PMID- 16220766 TI - Effect of spontaneous pregnancy reduction on obstetric outcome. AB - OBJECTIVE: To determine if obstetric outcome is compromised in pregnancies in which a spontaneous pregnancy reduction (SPR) occurred in the first trimester. STUDY DESIGN: Case-control study. RESULTS: First-trimester SPR was diagnosed in 29 (27.8%) of 104 twin pregnancies, 14 (28.6%) of 49 triplet pregnancies and 10 (28.6%) of 35 quadruplet pregnancies. Of these 53 patients, 15 were excluded from the analysis. In the remaining 38 women with SPR, vaginal bleeding occurred in 2 (5.3%) as compared to 7 (8.3%) of the controls. Pregnancy-induced hypertension occurred in 4 (10.5%) of SPR pregnancies as compared to 9 (10.7%) of control pregnancies. When compared to respective controls, there were no significant differences in the birth weights or gestational age at delivery of pregnancies spontaneously reduced to singletons (SPR, 38.5 weeks; controls, 38.2 weeks), twins (SPR, 36.2 weeks; controls, 34.4 weeks) or triplets (SPR, 31.0 weeks; controls, 32.0 weeks). CONCLUSION: SPR can be recognized in >25% of multiple pregnancies diagnosed in the early first trimester. Our data suggests that SPR is not associated with decreased gestational age at delivery, reduced birth weight or increased incidence of pregnancy-induced hypertension. PMID- 16220767 TI - Benefits of intracervical injection of sterile saline solution in laparoscopically assisted vaginal hysterectomy with vaginal colpotomy and bladder mobilization. AB - OBJECTIVE: To evaluate the influence of intracervical saline injection on inexperienced operators and on laparoscopically assisted vaginal hysterectomy (LAVH). STUDY DESIGN: This retrospective study included 273 women undergoing LAVH. From July 1997 to June 2002, 138 LAVHs were performed with laparoscopically approached colpotomies and bladder mobilization. Among 135 LAVHs with a vaginal approach, colpotomies/bladder mobilization was done directly in 62 and in the other 73 after a circumferential intracervical saline injection. All operations were performed by inexperienced operators under the supervision of senior surgeons. Blood loss, operative time and complications were analyzed. RESULTS: The average follow-up period was 41.2 +/- 17.4 months (range, 12-72). No statistically significant differences were observed in age, hemoglobin levels or length of postoperative hospital stay. The incidence of postoperative infection, hematoma and bowel injury was not significantly different. LAVH with vaginal colpotomies/bladder mobilization and intracervical saline injection was associated with the smallest estimated blood loss (p = 0.002) and operative time (p < 0.001). LAVH with laparoscopic colpotomies and bladder mobilization had the longest operative time (p<0.001) and the highest bladder injury rate (p= 0.004). CONCLUSION: A circumferential injection of normal saline at the cervicovaginal junction is a good option for inexperienced operators. PMID- 16220768 TI - Use of a levonorgestrel-releasing intrauterine device for the symptomatic treatment of uterine myomas. AB - OBJECTIVE: To assess the effect of a levonorgestrel-releasing intrauterine device (LNG-IUD) for the treatment of women with uterine myomas with increased bleeding by measuring total uterine volume and by determining patient clinical improvement, uterine artery flow velocity and hemoglobin levels. STUDY DESIGN: Ten patients with a complaint of increased uterine bleeding associated with the presence of uterine myomas were assessed in a descriptive case series and studied before and 6 months after placement of an LNG-IUD. The patients were assessed for intensity of bleeding, uterine volume, Doppler velocimetry of the uterine arteries, hematocrit and hemoglobin. RESULTS: One patient asked to be released from the study 15 days after LNG-IUD insertion, claiming the occurrence of increased bleeding, and 2 patients underwent spontaneous expulsion of the device after 2 and 4 months of follow-up. Of the 7 patients who continued the study, 3 started to present a bleeding pattern of the oligomenorrhea type, and the other 4 experienced amenorrhea. All patients (n = 6) with anemia at the beginning of the study presented normalization of hematocrit and hemoglobin levels after 6 months of treatment (p=0.0003 and p <0.0001, respectively). However, we observed no reduction in uterine volume (p = 0.11) or flow velocity assessed by the uterine artery pulsatility index (right, p=0.17; left, p=0.19) and of the series resistance index (right, p = 0.54; left, p =0.31) using Doppler velocimetry. CONCLUSION: Use of an LNG-IUD may be effective in controlling uterine bleeding secondary to myomatosis even in the absence of reduction of uterine volume. PMID- 16220769 TI - The physician as a factor in outpatient medical care after laparoscopically assisted vaginal hysterectomy: a multivariate analysis. AB - OBJECTIVE: To evaluate whether physician experience affects outpatient medical care utilization after laparoscopically assisted vaginal hysterectomy (LAVH). STUDY DESIGN: A total of 120 patients who underwent LAVH were included in the study; 84 underwent LAVH by 2 senior physicians, who had performed an average of 3 LAVHs per month for >6 years; 36 patients underwent LAVH by 3 junior physicians, who had performed an average of 1 case per month for < 3 years. Data were analyzed by the multiple linear regression model. The dependent variable was the number of outpatient visits within 3 months after discharge. The independent variables were patient age, parity, pelvic adhesions, chronic disease and physicians performing LAVH > 6 years. RESULTS: The patients in the 2 groups did not differ in age, parity, chronic disease or pelvic adhesions. The level of physician experience significantly influenced the mean number of outpatient visits after LAVH within 3 months. After controlling for patient characteristics, the mean number of outpatient visits after discharge within 3 months of LAVH performed by senior physicians with 6 years of experience was reduced (-1.98). CONCLUSION: LAVH performed by experienced physicians reduces the need for patients to seek postoperative outpatient medical care, thereby controlling medical costs incurred under the hospital global budget payment system. However, adding more variables to the multiple regression model, such as the socioeconomic status of patients, habits of seeking medical help, distance between home and hospital and relationship between patient and physician, may further explain the factors affecting outpatient medical care utilization. PMID- 16220770 TI - Puerperal group A Streptococcus infection: a case report. AB - BACKGROUND: Group A Streptococcus (GAS) sepsis is a rare event but carries a high risk of maternal mortality. CASE: A case of puerperal infection occurred with GAS. This patient had had an uneventful prenatal and intrapartum course. She was noted to have high, spiking fevers immediately postpartum, with minimal clinical symptoms. Her blood cultures were positive for GAS, most likely from a urinary tract infection. She was started on broad-coverage antibiotics and defervesced on postpartum day 4. She remained afebrile and was discharged on postpartum day 8. CONCLUSION: Patients with puerperal GAS sepsis commonly appear well clinically, with minor somatic complaints. GAS bacteremia should be suspected and promptly treated in women with high, spiking fevers early in the postpartum period. There are currently no guidelines on preventing vertical transmission. It is unclear how a patient with a previous history of GAS should be managed. Prophylactic use of penicillin during future labor may be warranted. PMID- 16220771 TI - Incarceration of a retroflexed, gravid uterus from severe uterine prolapse: a case report. AB - BACKGROUND: Acute urinary retention as a result of incarceration of a retroflexed, gravid uterus is a known phenomenon. However, prolapse as a risk factor has not been previously described. CASE: A 40-year-old woman, gravida 4, para 2, with an intrauterine gestation of 19 weeks presented to the emergency room complaining of inability to void for the previous 12 hours and difficulty voiding and constipation for the previous 6 weeks. She had a history significant for stage III uterine prolapse in early pregnancy. Foley catheterization yielded 800 mL of urine, and an examination revealed a retroflexed uterus. The cervix was displaced anteriorly behind the pubic symphysis. Ultrasound confirmed these findings and the presence of a viable gestation. The uterus was successfully manually displaced under epidural anesthesia. The patient was able to void without difficulty after uterine displacement. CONCLUSION: Incarceration of a retroflexed uterus should be considered in the differential diagnosis in any woman who presents with voiding difficulty in the late first or second trimester. Uterine prolapse is a risk factor for incarceration of a retroflexed uterus. Epidural anesthesia should be considered for a patient if manual uterine displacement cannot be performed successfully without anesthesia. PMID- 16220772 TI - Ovarian abscess caused by nontyphoidal Salmonella in a woman with systemic lupus erythematosus: a case report. AB - BACKGROUND: Nontyphoidal Salmonella infection in a patient with systemic lupus erythematosus (SLE) is rare. CASE: A 24-year-old woman with a 9-year history of SLE, on corticosteroids, presented with severe abdominal pain, fever, vomiting and diarrhea for 1 day. Transabdominal sonography and abdominal computed tomography showed a right-sided well-defined pelvic mass. Laboratory study revealed leukocytosis. An emergency laparotomy revealed a right ovarian mass with purulent exudate. A right partial oophorectomy was performed. Pathologic examination revealed a hemorrhagic ovarian cyst with focal abscess. Culture of the ovarian mass grew Salmonella typhimurium. Postoperatively the patient did well and was discharged 5 days later. CONCLUSION: Patients with SLE are at risk of Salmonella infection. Gynecologists should be aware of the possibility of an ovarian abscess caused by Salmonella in such patients presenting with a pelvic mass and fever. PMID- 16220773 TI - Leiomyosarcoma of the vulva: a case report. AB - BACKGROUND: Leiomyosarcoma of the vulva is a rare gynecologic malignancy, comprising approximately 1% of vulvar cancers. CASE: A 36-year-old woman was referred for a slowly growing painless vulvar mass that was initially thought to be a Bartholin's duct cyst but was cancerous upon biopsy. A modified radical vulvectomy was performed, and pathology revealed a grade 1 leiomyosarcoma. Thirteen months later, the tumor had not recurred. CONCLUSION: Vulvar cancer must be considered in patients with a suspected Bartholin duct cyst that demonstrates atypical features. A biopsy should be obtained if the mass appears firm or solid on palpation, is ulcerated or presents in a slightly different location from the usual area of the Bartholin gland. PMID- 16220774 TI - Fibrolamellar hepatocellular carcinoma arising in a background of focal nodular hyperplasia: a report of 2 cases. AB - BACKGROUND: Fibrolamellar carcinoma, a rare variant of hepatocellular carcinoma, and focal nodular hyperplasia, a benign lesion, are rare hepatic lesions that are known to occur in young women with noncirrhotic livers. Some have suggested that fibrolamellar carcinoma might be the malignant counterpart of focal nodular hyperplasia. The coexistence of the 2 lesions is very rare. CASES: Two cases of fibrolamellar hepatocellular carcinoma arising in a background of focal nodular hyperplasia followed long-term oral contraception, and 1 of the 2 occurred during pregnancy. CONCLUSION: Distinguishing fibrolamellar carcinoma from focal nodular hyperplasia has important implications for treatment and prognosis. One should be aware of such conditions, especially in patients with a long history of oral contraception. PMID- 16220775 TI - Dichorionic twins discordant for thanatophoric dysplasia managed with selective reduction at 20 weeks' gestation: a case report. AB - BACKGROUND: Thanatophoric dysplasia (TD) is a rare and lethal form of skeletal disorder. A MEDLINE search for 1965-2003 yielded only 3 reports of multiple pregnancies discordant for TD. This is the first case report of selective twin reduction for this diagnosis. CASE: A young woman was seen in consultation at 20 weeks' gestation. Ultrasound examination revealed a twin pregnancy, with ultrasound markers consistent with thanatophoric dysplasia, type II, in twin A. A thick dividing membrane and separated placentas were noted. After counseling, the patient opted for selective termination of twin A. Termination was performed by intracardiac injection of potassium chloride. The pregnancy continued uneventfully until 33 weeks, when spontaneous labor resulted in vaginal delivery of a vigorous female infant, and a mummified, macerated fetus. CONCLUSION: Selective termination for discordant lethal anomalies can be safely performed when the presence of the anomalous twin increases the risk of a poor perinatal outcome for the apparently normal cotwin. PMID- 16220776 TI - Intraepithelial mucinous carcinoma arising in an endocervical-type mucinous polypoid adenomyoma of the uterine corpus: a case report. AB - BACKGROUND: Benign endocervical-type mucinous adenomyoma arising in the uterine corpus is a rare entity. We report a case of intraepithelial mucinous adenocarcinoma arising in an endocervical-type mucinous adenomyoma of the uterine corpus in a previously healthy woman. CASE: A 55-year-old, white woman presented with postmenopausal bleeding. Uterine leiomyomas and endometrial polyps were suggested by ultrasonography. Endometrial curettage showed multiple polypoid tissue fragments, each composed of glands lined with tall columnar endocervical type mucinous epithelium lying within a background of endometrial-type stroma and smooth muscle fibers. The mucinous glandular epithelium showed a spectrum of architectural and cytologic changes ranging from benign to severe atypia and occasional back-to-back cribriform glands, consistent with adenocarcinoma. The surrounding smooth muscle fibers and endometrial stromal cells were benign. A diagnosis was made of intraepithelial adenocarcinoma arising in a mucinous adenomyoma in the uterine corpus. CONCLUSION: Endocervical-type mucinous adenomyoma, although previously reported as a benign entity, may contain areas of adenocarcinoma. PMID- 16220777 TI - [Nootropic mechanisms of ginsenoside Rg1--influence on neuronal plasticity and neurogenesis]. PMID- 16220778 TI - [Current status of HIV protease inhibitors]. PMID- 16220779 TI - [Difficulty and hot-points on pharmacokinetics studies of traditional Chinese medicine]. PMID- 16220780 TI - [Vasodilation effect of atropine on rat mesenteric artery]. AB - AIM: To study the vasodilation effect of atropine and its mechanism. METHODS: Isometric tension was recorded in isolated rat super mesenteric arteries precontracted by noradrenaline (NE) to study the vasodilation effect of atropine, and to investigate the role of endothelial cell and vascular smooth muscle cell on vasodilation. RESULTS: Atropine was shown to significantly dilate the endothelium-intact and endothelium-denuded arteries precontracted by NE. Nomega Nitro-L-arginine methyl ester (L-NAME, nitric oxide synthase inhabitor), indomethacin (cyclooxygenase inhibitor), propranolol (general beta adrenoceptor antagonist) and glibenclamide (ATP sensitive potassium channel inhibitor) showed no effect on vasodilation of atropine. Atropine did not affect the concentration contraction curve of K+. However, atropine suppressed the contraction induced by NE and CaCl2, but not that by caffeine in the Ca+ -free Krebs solution. CONCLUSION: Atropine showed significant vasodilation effect which may derive, in part, from endothelium. Besides, atropine could inhibit the receptor-mediated Ca2+ -influx and Ca2+ -release, which was inferred to the mechanism of atropine on vasodilation. PMID- 16220782 TI - Effects of ferulic acid on E-selectin expression in activated endothelial cell and leukocyte-endothelial cell adhesion. AB - AIM: To study the effects of ferulic acid (FA) on E-selectin expression in human umbilical vein endothelial cells (HUVECs) activated by lipopolysaccharide and leukocyte-endothelial cell adhesion. METHODS: The effects of FA on E-selectin and E-selectin mRNA expression were determined by flow cytometry and reverse transcription polymerase chain reaction. The effect of FA on HL60-HUVEC adhesion was evaluated with the method of staining the cells by Rose Bengal. RESULTS: The expression of E-selectin and E-selectin mRNA were down regulated by FA (0.62 and 0.41 mmol x L(-1), respectively). HL60 cells adhered to activated HUVECs were also reduced by FA (0.62 and 0.41 mmol x L(-1), respectively). CONCLUSION: FA can inhibit the expression of E-selectin and E-selectin mRNA and HL60-HUVEC adhesion. This may contribute to its protective effect against ischemia-reperfusion injury. PMID- 16220781 TI - [Effect of proanthocyanidins on COX-2 enzyme activity and COX-2 mRNA /protein expression in LPS-induced RAW264.7 cells]. AB - AIM: To study the effect of proanthocyanidins on the COX-2 enzyme activity and COX-2 protein expression in LPS-induced RAW264.7 cells. METHODS: After being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 mg x L(-1) LPS for 9 h, the effect of proanthocyanidins on the activity of COX-2 enzyme in RAW264.7 cells was analysed by radioimmunoassay (RIA). After being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 mg x L(-1) LPS for 9 h, the effects of proanthocyanidins on the expressions of COX-2 mRNA and protein in RAW264.7 cells were analysed by RT-PCR and Western blotting. RESULTS: The activity of COX-2 enzyme was not inhibited by proanthocyanidins (0. 8, 4 and 20 mg x L(-1), P > 0.05 vs LPS group), but the activity of COX-2 enzyme was significantly inhibited by 10 micromol x L(-01) NS 398 (P < 0.01 vs LPS group). The expression of COX-2 mRNA was inhibited by proanthocyanidins (0. 8, 4 and 20 mg x L(-1)). The expression of COX-2 protein was inhibited by proanthocyanidins (4 and 20 mg x L(-1)). CONCLUSION: Proanthocyanidins had no effect on the activity of COX-2 enzyme in LPS-induced RAW264. 7 cells. Proanthocyanidins inhibited significantly the expression of COX 2 mRNA and protein in LPS-induced RAW264.7 cells. PMID- 16220784 TI - [Establishment of an IR-HIRc cell model for screening GFAT inhibitor]. AB - AIM: To set up an IR-HIRc cell model for screening the inhibitor of GFAT (glutamine: fructose-6-phosphate amidotransferase) , the key enzyme in the hexosamine biosynthesis pathway (HBP). METHODS: For GFAT activity assay, the GDH method was improved by adjusting the value of pH in the reaction system and the concentrations of the reactants. The sensitivity to insulin in the cells was estimated by the measurement of insulin-induced glucose-uptake. The IR-HIRc model was set up by the stimulation of long-action insulin for 36 h. The IR-HIRc model and GDH method was used for screening GFAT inhibitor. RESULTS: With the administration of 25 nmol x L(-1) long-action insulin in HIRe cells for 36 hours, the GFAT activity increased by 47% and the insulin-induced glucose-uptake decreased by 21%. Azaserine, a GFAT inhibitor, inhibited GFAT activity significantly in a dose-dependent manner in IR-HIRc model. CONCLUSION: With the stimulation of 25 nmol x L(-1) long-action insulin for 36 h, excess hexosamine flux and insulin resistant in IR-HIRc cell model was set up, which can be used for screening PMID- 16220783 TI - [Intervention of cetirizine on monocyte chemoattractant protein-1 in cutaneous inflammation]. AB - AIM: To study the intervention of cetirizine on monocyte chemoattractant protein 1 (MCP-1) in different cutaneous inflammation models. METHODS: Histamine and IFN gamma stimulated dermal fibroblast cells and HaCaT cells to mimic cutaneous inflammation. Expression of MCP-1 was assessed by means of RT-PCR and ELISA. RESULTS: Compared with the control group of dermal fibroblast (DF) cells and HaCaT cells, MCP-1 mRNA was significantly upregulated by histamine (10 micromol x L(-1)) and IFN-gamma (20 ng x mL(-1)). The protein secretions of MCP-1 were increased 3.5 fold and 8.4 fold in DF cells, respectively. The similar tendency was observed in HaCaT cells. The enhancing effects of histamine and IFN-gamma on MCP-1 protein production were significantly inhibited by cetirizine (1 and 10 micromol x L(-1)) in DF and HaCaT cells. CONCLUSION: Cetirizine may exert the anti-inflammatory effect of skin via inhibiting MCP-1 expression. PMID- 16220785 TI - [Synthesis of derivatives of (9S)-12-methylene erythromycin and their antibacterial activity in vitro]. AB - AIM: To synthesizs of derivatives of (9S)-12-methylene erythromycin possessed potent antibacterial activity. METHODS: Using erythromycin A as a starting material, via two intermediate compounds protected 12,21-dehydroerythromycin A and 6,7: 12,21-didehydro erythromycin A, several 9-O, 11-O-ethylidene compounds were obtained. During this process, benzyl and isopropyl have been selected as the protecting group. The structures of compounds obtained were confirmed with 13C NMR and MS-FAB. Their antibacterial activity in vitro was tested. RESULTS: Eleven derivatives of erythromycin were synthesized. Five of them were unknown compounds. CONCLUSION: The preliminary biological test showed that two target compounds exhibited less potent antibacterial activity in vitro. PMID- 16220786 TI - [Studies on lignan glycosides from the roots of Bupleurum scorzonerifolium]. AB - AIM: To study the lignan glycosides from the roots of Bupleurum scorzonerifolium. Chromatography methods was used to isolate compounds and chemical and spectral methods were used to identify the structures of isolated compounds. RESULTS: Two lignan glycosides were isolated from the roots of B. scorzonerifolium and their structures were identified as 2, 3-E-2,3-dihydro-2-(3'-methoxy-4'-O-beta-D glucopyranosyl-phenyl)-3-hydroxymethyl-5-(3"-hydroxypropenyl )-7-methoxy-1-benzo [b] furan (1) and 2, 3-E-2, 3-dihydro-2-( 3'-methoxy-4'-hydroxy-phenyl)-3 hydroxymethyl-5-( 3"-hydroxypropenyl)-7-O-beta-D-glucopyranosyl-1-benzo[b]furan (2), both of them which were the mixture of (+)2S,3R- and (-)2R,3S-type diastereoisomer and in both of them the amount of (+)2S,3R-type was a little more than that of the (-)2R,3S-type. CONCLUSION: Compounds 1 and 2 were isolated from B. scorzonerifolium for the first time. (+)2S, 3R-2, 3-Dihydro-2-(3'-methoxy-4' hydroxy-phenyl)-3-hydroxymethyl-5-(3"-hydroxypropenyl)-7-O-beta-D-glucopyranosyl 1-benzo[b] furan is a new compound. PMID- 16220787 TI - [The chemical constituents of Knoxia valerianoides]. AB - AIM: To study chemical constituents of Knoxia valerianoides Thorel et Pitard. METHODS: Chromatographic methods were used for the isolation and purification. Structures was elucidated on the basis of chemical analysis and spectroscopic data. RESULTS: Two anthraquinone were isolated from Knoxia valerianoides Thorel et Pitard and identified as 1,3,5-trihydroxy-2-ethoxymethyl-6-methoxyl anthraquinone (I) and 1,3-dihydroxy-2-ethoxymethyl-anthraquinone (II). CONCLUSION: The compound I was found to be a novel anthraquinone constituent and II was isolated from Knoxia valerianoides Thorel et Pitard for the first time. PMID- 16220789 TI - Three biflavonoids from ethanol extract of the root of Daphne genkwa. AB - AIM: To investigate the chemical constituents of the secondary metabolites of the roots of Daphne genkwa. METHODS: The roots of D. genkwa were extracted with 95% ethanol at 60-70 degrees C for 7 days to obtain the crude extract. The crude extract was purified by silica gel and Sephadex LH-20 column chromatography as well as the HPLC techniques. The structures of the isolates were elucidated by combined spectroscopic methods including 1D and 2D NMR, MS, UV, IR and CD. RESULTS: Three new biflavonoids were isolated from the ethanol extract of the roots of D. genkwa and their structures were identified as daphnodorin H-3-methyl ether (1), daphnodorin H-3"-methyl ether (2) and daphnodorin G-3"-methyl ether (3). CONCLUSION: Compounds 1, 2 and 3 are three new biflavonoids. PMID- 16220788 TI - [Studies on chemical constituents of Pinus armandii]. AB - AIM: To study chemical constituents from pine cone of Pinus armandii Franch. METHODS: The constituents were isolated by chromatographic method and the structures were identified on the basis of spectral analysis. RESULTS: Four compounds were identified as 7-oxo-12alpha, 13beta-dihydroxyabiet-8(14)-en18-oic acid (I), 7-oxo-13beta-hydroxyabiet-8 (14)-en-18-oic acid (II), 8 (14)-podocarpen 13-on-18-oic acid (III) and lambertianic acid (IV). CONCLUSION: Compound I is a new diterpenoid and compounds II, III were isolated from this plant for the first time. PMID- 16220790 TI - A new flavanone from Dryopteris sublaeta. AB - AIM: To study the chemical constituents of Dryopteris sublaeta Ching et Hsu. METHODS: Fresh plant of Dryopteris sublaeta Ching et Hsu was extracted twice with boiling water, the extract was concentrated to small volume under reduced pressure at 50 degrees C. The concentrated material was partitioned with ether, ethyl acetate, and n-butanol. The fraction of ether extract was chromatographed over silica gel column. The compounds were identified on the basis of their physiochemical and spectral data. RESULTS: Four compounds were obtained and identified as 2 (S)-5, 7, 3'-trihydroxy-6, 8-dimethyl-5'-methoxyflavanone (1), matteucinol (2), desmethoxymatteucinol (3) and 5, 7, 2'-trihydroxy-6, 8 dimethylflavanone (4). CONCLUSION: Compound 1 is a new one, the others were isolated from Dryopteris for the first time. PMID- 16220791 TI - [HPLC fingerprinting of total glycosides of Swertia franchetiana]. AB - AIM: To establish a sensitive and specific HPLC method for controlling the quality of total glycosides from Swertia franchetiana H. Smith. METHODS: HPLC method was applied for quality and quantitative assessment of the pharmaceutical extracts from Swertia franchetiana H. Smith. The preparation of sample, the HPLC column, mobile phase, elution mode (isocratic or gradient) and gradient program were optimized in order to obtain HPLC profile. The HPLC system consisted of a SPD-1OAvp pump, SPD-M1OAVP photodiode-array detector (PAD), SIL-10ADVP auto injector. Data were acquired and processed with the CLASS-VP6.1 workstation. HPLC analysis was performed on a Kromasil C18 column (250 mm x 4. 6 mm ID, 5 microm) with methanol and water as mobile phase. The column temperature was set up at 40 degrees C and the flow-rate was 1 mL x min(-1). The reference solution of chemical standards and sample were injected into HPLC system, separately. RESULTS: The HPLC chromatographic fingerprinting of the total glycosides, showing 16 characteristic peaks which were partitioned into three parts: one peak in 0-10 min of retention time, nine peaks containing main 1-7 peaks in 10-15 min of retention time, 6 peaks in 15-30 min of retention time, was established from 10 lots of their products. By comparison of the retention time and the on-line UV spectra and their molecule weights of chemical standards, peak 1-7 were identified as swertiamarin (1), gentiopicroside (2), sweroside (3), isoorientin (4), swertisin (5), isoswertisin (6) and swetianolin (7), respectively. The ratios of peak area between 1-16 were in their extent. Moreover, comparison of the HPLC profiles of the total glycosides, the extracts prepared using another process and the plant indicated that they were closely related to each other. CONCLUSION: The HPLC profiles and quantitative assessment of the total glycosides from Swertia franchetiana H. Smith with high specificity can be used to control their quality and assure lot to lot consistency. PMID- 16220792 TI - [Tissue distribution and excretion of bromotetrandrine in rats]. AB - AIM: To study the tissue distribution and excretion of bromotetrandrine (W198) in rats. METHODS: The concentrations of W198 in biological samples were determined by an HPLC method with UV detection. RESULTS: After a single i.v. dose of 20 mg x kg(-1) W198 in rats, the parent drug concentrations in tissues were higher than those in blood at the same time. Parent drug was mainly distributed in lung, kidney, heart and liver, the peak levels were attained at 0.25 h and decreasing at 2 h after dosing in most tissues. After a single iv dose of 20 mg x kg(-1) W198 in rats, the excretion of the parent drug in urine, feces and bile amounted to 0. 150%, 2.1% and 0.063% of the dose, respectively. CONCLUSION: W198 was mostly distributed in lung. The parent drug excretion was less than 3% via urine, feces and bile. PMID- 16220793 TI - [The pharmacokinetics and bioequivalence of acipimox sustained-release tablets after a single and multiple oral administration in healthy dogs]. AB - AIM: To study the pharmacokinetics and bioequivalence of acipimox sustained release tablets (SRT) after a single and multiple oral dose in healthy dogs. METHODS: The plasma concentrations of of SRT and reference capsules with a single and multiple oral doses. RESULTS: The drug concentration-time profiles fitted to a noncompartment model. After a single dose administration of sustained-release tablets and capsules, the pharmacokinetic parameters were as follows: AUC were (158 +/- 30) and (147 +/- 37) microg x h x mL(-1); Tmax were (4.3 +/- 0.8) and (2.6 +/- 1.3) h; Cmax were (29 +/- 6) and (42 +/- 10) microg x mL(-1); T(1/2) were (2.3 +/- 0.7) and (1.60 +/- 0.10) h; MRT were (6.0 +/- 0.8) and (3.9 +/- 0.7) h, respectively. The relative bioavailability of the sustained-release tablet was (108 +/- 16) %. After a multiple oral administration of sustained release tablets and capsules, the pharmacokinetic parameters were as follows: AUC were (209 +/- 23) and (195 +/- 26) microg x h x mL(-1); Tmax were (6.3 +/- 0.8) and (3.4 +/- 1.5) h; Cmax were (27 +/- 4) and (36 +/- 5) microg x mL(-1); Cmmin were (2.2 +/- 1.0) and (0.20 +/- 0.20) microg x mL(-1); Cav were (8.7 +/- 1.0) and (8.1 +/- 1.1) micro x mL(-1); FI were (293 +/- 73) % and (448 +/- 91) % , respectively. The relative bioavailability of the sustained-release tablet was (114 +/- 19) %. CONCLUSION: The results of two one-side test from single dose administration shown that two preparations were bioequivalent. The Cmax of sustained-release tablet was lower than that of capsules, while the Tmax and MRT of sustained-release tablet were higher than that of capsule, which indicating a good retarding effect. The results from multiple dose administration also shown that two preparations were bioequivalent and the DF of sustained-release tablet was significant lower than that of capsule. PMID- 16220794 TI - [Preparation of nosiheptide liposomes and its inhibitory effect on hepatitics B virus in vitro]. AB - AIM: To prepare liposomes of nosiheptide and study its ability to inhibit hepatitis B virus HBsAg and HBeAg secreted. METHODS: Liposomes of nosiheptide was prepared by sodium deoxycholate dialysis and sonication. Nosheptide was determined by HPLC and partical size was determined by using laser light scattering instrument. Transmission electron microscopy (TEM) was used to examine the morphology of liposomes. Its actions to inhibit hepatitis B virus HBsAg and HBeAg secreted was studied by a HBV-transfectted cell line (HepG2 2. 2. 15 ). RESULTS: Encapsulation efficiency of liposomes by chloroform:methanol (2:1, v/v) was higher than that by dioxane. With the increase of the ratio of nosiheptide: PC (W/W), the encapsulation efficiency of liposomes decreased with the increase of ratio of sodium deoxycholate: PC, the liposomes partical size decreased. The liposomes kept stable at -20 degrees C after 2 years. The drug concentrations of liposomes that inhibit HBsAg secreted by (46.9 +/- 2. 6) %, (55.4 +/- 1.2) %, (65 +/- 3) % and HBeAg secreted by (15.1 +/- 2.3) %, (36.2 +/- 1.7) %, (36.8 +/- 2.5) % were 1.25, 2.5, 5.0 microg x mL(-1), respectively. CONCLUSION: Liposomes of nosheptide can be prepared by sodium deoxycholate dialysis and sonication, which ability to inhibit hepatitis B virus HBsAg and HBeAg secreted is better than nosheptide. PMID- 16220795 TI - Effects of intranasal administration of nimodipine on cerebral hemodynamics of dogs. AB - AIM: To investigate the effect of nimodipine (NM) on cerebral blood flow (CBF) in dogs following intranasal administration. METHODS: NM solution was administered intranasally, intravenously (i.v.), and orally to dogs and the change of CBF was determined by using electromagnetic blood flowmeter. MFLab experimental program was applied to monitor the experimental process and analyze data. RESULTS: CBF markedly increased after iv and intranasal application, while large variance was observed after oral dosing. CBF in dogs after three administrations increased by 26.4%, 28.0% and 8.5%, respectively, compared with that of baseline. Following intranasal administration, the onset of action was slightly slower than that after iv injection [(5 +/- 4) min vs (2.2 +/- 1.2) min], however the duration of improvement was the longest [ (25 +/- 17) min]. CONCLUSION: Intranasal delivery for NM can be a promising alternative to parenteral or oral administration. PMID- 16220797 TI - [The in vitro cytotoxicity and in vivo toxicity of doxorubicin antiresistant stealth liposomes]. AB - AIM: Multidrug resistance ( MDR) as a major obstacle to successful clinical cancer chemotherapy, searching a novel effective antiresistant drug would be necessary. METHODS: A novel doxorubicin anti-resistant stealth liposomes (DARSLs) was prepared by co-encapsulating doxorubicin (DOX) and verapamil (VER) into stealth liposomes with ammonium sulfate gradient remote loading approach. In vitro cytotoxity of various DOX formulations and in vivo toxicity of DARSLs were evaluated using DOX-resistant rat prostate cancer cell line (MLLB2), human uterus sarcoma cell line (MES-SA/DX5) and normal SD rats, separately. RESULTS: The DARSLs liposome suspensions mainly consisted of homogeneous large unilamellar vesicles (LUV) with average particle size of (118.1 +/- 22.3) nm. Encapsulation efficiencies of DOX and VER in DARSLs were more than 90% and about 70%, respectively, when the ratio of DOX/VER/Lipid was 1: 0.11 :10 (w/w/w). In vitro cytotoxicity tests of the DARSLs using rat prostate cancer cell line (MLLB2) and human uterus sarcoma cell line (MES-SA/DX5) showed that 5 micromol x L(-1) VER significantly reversed DOX-resistance of these 2 cell lines and DARSLs was the most effective on inhibition of DOX-resistant cell growth. Besides, compared to FDFV, much slower DOX distribution (confocal microscopy) to nuclei and cytoplasm in MLLB2 cells for DARSLs suggested that it might possess distinct mechanism of cytotoxicity. Systemic and cardiac toxicity evaluations in normal SD rats suggested that liposomal encapsulation could significantly improve the severe cardiotoxicity arising from simultanous administration of DOX and VER. CONCLUSION: DARSLs is a novel anticancer liposome formulation with lower cardiotoxicity, effective drug-resistance reversal and intravenous injection. PMID- 16220796 TI - [Isopropyl myristate molecular gels and drug-loaded transdermal capability]. AB - AIM: To prepare of isopropyl myristate (IPM) molecular gels and investigate of its transdermal capability. METHODS: Microstructure of IPM gels was studied by scanning electron microscope (SEM) and optical microscope (OM). The rheology and thixotropy of IPM gels were investigated by viscosity. Triptolide was used as model drug to investigate its transdermal capability. RESULTS: The microstructure of IPM gels was a three-dimension network formed by the aggregation of Span 60 in IPM, which was rod-like tubular aggregate. It has good rheology and thixotropy. There was a good linear correlation between the accumulative permeated amount per unit area and the time for triptolide-loaded IPM gels. The permeation process agreed with zero order pharmacokinetics. The average permeability through rat skin for triptolide was 19.26 ng x cm(-2) x h(-1), which was 2.92 times of triptolide unguents obtained commercially available. CONCLUSION: Isopropyl myristate molercular gel can be formed by span 60 assemblies. Transdermal capability drug-loaded IPM gels was better than that of triptolide unguents. PMID- 16220798 TI - Meetings and a goodbye. PMID- 16220799 TI - Primus inter pares: William R. Laney, DMD, MS. Interview by George Zarb. PMID- 16220800 TI - Long-span, fixed-movable, resin-bonded fixed partial dentures: a retrospective, preliminary clinical investigation. AB - PURPOSE: Long-span, resin-bonded fixed partial dentures (FPDs) have been associated with higher debonding rates than short-span prostheses. The use of modified nonrigid connectors that allow movement between the abutments in long span resin-bonded prostheses may reduce harmful interabutment forces that stress the metal framework and resin-bonded interface. This preliminary investigation aimed to evaluate the longevity of long-span resin-bonded FPDs of 4 or more units with a modified nonrigid connector and increased extension of the retainer framework around the major abutment. MATERIALS AND METHODS: Thirty-eight patients attended a clinical examination; each had been provided with 1 or more resin bonded FPD of 4 or more units (43 prostheses). For each patient, the following data were recorded: gender, age, cementation date, and endodontic treatment, if performed. Data regarding the occurrence of any debondings and patient expectations were also recorded. RESULTS: The mean service life for the 43 prostheses was 46.9 months (SD 22.0), with a range of 12 days to 87 months. Three prostheses had debonded, resulting in a clinical retention rate of 92.2%. CONCLUSION: Long-span resin-bonded FPDs incorporating nonrigid connectors that allow independent movement between the major and minor retainer, combined with increased framework extension on the major abutment, appear successful in the short term. Further research is required to determine their long-term efficacy. PMID- 16220801 TI - Increased antibacterial activity of zinc polycarboxylate cement by the addition of chlorhexidine gluconate in fixed prosthodontics. AB - PURPOSE: This study evaluated the antibacterial activity of water-activated zinc polycarboxylate cement with adjunctive 0.12% chlorhexidine gluconate on the subgingival microbiota in fixed partial dentures. MATERIALS AND METHODS: Thirty six teeth prepared as fixed partial denture abutments in 9 patients were cemented randomly using water-activated zinc polycarboxylate cement (control group) or water-activated zinc polycarboxylate cement, including 0.12% chlorhexidine gluconate (test group). A total of 108 subgingival plaque samples were analyzed at baseline, immediately before permanent cementation (5 weeks), and 8 weeks later (at 13 weeks). RESULTS: In the control group, the subgingival microbiota altered to closely resemble the flora of chronic gingivitis (increased proportions of gram-negative anaerobes such as Prevotella intermedia, Fusobacterium nucleatum) by 13 weeks. In contrast, the microflora at test sites comprised predominantly gram-positive facultative cocci and rods at 13 weeks. CONCLUSION: This study demonstrated that the addition of 0.12% chlorhexidine gluconate may enhance the antimicrobial action of polycarboxylate cements to ensure the maintenance of a microflora compatible with periodontal health, at least up to 13 weeks post-cementation. PMID- 16220802 TI - Implant-prosthodontic treatment for special care patients: a case series study. AB - PURPOSE: The aim of the study was to assess implant survival and complications with implants and prostheses in patients exhibiting a variety of systemic diseases and congenital defects. MATERIALS AND METHODS: Patients with specific medical conditions are regularly treated at the Department of Prosthodontics, University of Bern, Switzerland. All those who had received implant-prosthodontic treatment during the past 12 years were reexamined for this study. Among these patients the following diseases were observed: cleft lip/palate (n = 8), Down syndrome (n = 3), Sjogren syndrome and scleroderma (n = 2), ectodermal dysplasia (n = 4), developmental retardation (n = 2), chronic leukemia (n = 2), lichen planus (n = 1), cerebral palsy (n = 1), deaf-muteness (n = 1), amyotrophic lateral sclerosis (n = 1). At the time of the treatment the mean age was 55.6 years. ITI implants had been placed according to a standard protocol with local anesthesia, except for one patient in whom full anesthesia was used. One hundred three implants were loaded and supported a total of 34 fixed or removable prostheses. All patients were appointed to a regular maintenance care program. In the context of the present study, all but 1 patient were reexamined clinically. New radiographs were obtained, and the implants and prostheses assessed. Additional information was obtained from regular records in the patients' charts. RESULTS: Three implants were lost in the healing phase, and 1 implant was replaced. Only 1 patient with 4 implants was lost from the study (she had passed away). The survival rate of the loaded implants was 100%. In 1 patient, peri implant bony defects were detected around all 3 intraforaminal implants. The prosthetic plan was maintained in all patients, and they continued to wear the originally planned type of prosthesis. Complications included insufficient hygiene, soft tissue hyperplasia, extraction of remaining teeth, and minor maintenance or repair of the prostheses. CONCLUSIONS: So far, the mostly unknown implications and possible risks for the process of osseointegration and long-term maintenance in patients with such rare diseases and defects has resulted in a rather restricted application of implants. However, from the present results, it appears that implants can successfully be placed and maintained. This is ascribed in part to a strict maintenance care program provided by the caregivers and to a high compliance of the patients who participated in this program to perform good oral hygiene. PMID- 16220804 TI - In vitro study of the adherence of Candida albicans to acrylic resins: relationship to surface energy. AB - PURPOSE: This study aimed to determine whether small variations in the composition of the polymethylmethacrylate (PMMA) of widely used dentures produce differences in the degree of Candida albicans adherence and to relate any differences found to the surface energy of the resins, which appears to play a major role in the initial phases of microorganism adhesion. MATERIALS AND METHODS: A reference strain of C albicans (18.804 ATCC) and 11 different PMMAs (Vacalon, Inkotherm 85, Veracril, Probase Cold, Inkotherm Press, Inkotherm 85 T, Ruthinium, Vertex, SR Ivocap, Idoacryl, Lucitone) were used. Fifty specimens (15 x 10 x 1 mm) of each type were prepared. C albicans adhesion was determined by microorganism count under fluorescent optical microscope, and the surface energy of the resins was calculated by the contact angle method. P < .05 was regarded as significant. RESULTS: C albicans adhesion on the resins ranged from 7.12 cells/mm2 to 330.8 cells/mm2, with statistically significant (P > .05) differences in some cases. Despite small variations in the composition of the resins, their surface energy values were very similar (38.78 to 41.2 mJ/m2), and no relationship was found between C albicans adhesion and surface energy. CONCLUSION: The adhesion of C albicans to different resins varied in vitro, possibly as a result of the action of residual postpolymerization products. According to these results, variations in surface energy that result from differences in the composition of the different PMMA resins appear to have no influence on the adhesion of C albicans or, therefore, on the onset of denture stomatitis. PMID- 16220803 TI - Marginal fit of titanium metal-ceramic crowns. AB - PURPOSE: This study aimed to compare the marginal fit of 2 kinds of metal-ceramic crowns-crowns cast from commercially pure titanium and Procera titanium crowns. MATERIALS AND METHODS: Ten copings of each type were prepared, veneered with low fusing porcelain, and bonded with glass-ionomer cement. Marginal fit was assessed before and after cementation, and the data were analyzed statistically. RESULTS: There were significant differences among mean values of marginal fit between the groups. Cementation increases discrepancies in both groups. CONCLUSION: Casting titanium has resulted in the highest discrepancies in marginal fit of both groups. PMID- 16220805 TI - Clinical evaluation of the use of fiber posts and direct resin restorations for endodontically treated teeth. AB - PURPOSE: Restoration of root-treated teeth is routinely performed in clinical practice with a choice of therapeutic options, considering many factors to provide optimal mechanical properties, esthetics, and longevity. The aim of the present work was to present a preliminary clinical report on the use of fiber posts and direct resin composites for restoring root-treated teeth. MATERIALS AND METHODS: Thirty-eight anterior and 62 posterior endodontically treated teeth were selected from 3 private prosthodontic offices. The protocol used included endodontic treatment, with translucent fiber posts (DT post) bonded to the post space using a '1-bottle' adhesive (One-Step, Bisco) and a dual-cure resin cement (DuoLink, Bisco). Direct resin restorations were performed using a micro-hybrid resin composite (Gradia Direct, GC) and a layering technique. Both opaque dentin and enamel and translucent enamel shades were used. RESULTS: Patients were recalled after 6, 12, 24, and 30 months, and the restorations assessed according to predetermined clinical and radiographic criteria. These clinician-mediated evaluation methods confirmed the good clinical performance of the restorations. CONCLUSION: Restoration of endodontically treated teeth with fiber posts and direct resin composites is a treatment option, that in the short term conserves remaining tooth structure and results in good patient compliance. PMID- 16220806 TI - A comparison between computerized tomography, magnetic resonance imaging, and laser scanning for capturing 3-dimensional data from an object of standard form. AB - PURPOSE: The study's aim was to compare dimensional measurements on computer images generated from data captured digitally by 3 different methods of the surfaces of a plastic cube of known form to those obtained directly from the cube itself. MATERIALS AND METHODS: Three-dimensional images were reconstructed of a plastic cube obtained by computerized tomography (CT), magnetic resonance imaging (MRI), and laser scanning. Digital calipers were used to record dimensional measurements between the opposing faces of the plastic cube. Similar dimensional measurements were recorded between the cube faces on each of the reconstructed images. The data were analyzed using a 2-way ANOVA to determine whether there were differences between dimensional measurements on the computer images generated from the digitization of the cube surfaces by the different techniques, and the direct measurement of the cube itself. RESULTS: A significant effect of how the measurements were taken (ie, direct, CT, MRI, and laser scanning) on the overall variation of dimensional measurement (P < .0005) was observed. Post hoc tests (Bonferroni) revealed that these differences were due principally to differences between the laser-scanned images compared to other sources (ie, direct, CT, and MRI). The magnitude of these differences was very small, up to a maximum mean difference of 0.71 mm (Cl +/- 0.037 mm). CONCLUSION: All 3 methods of imaging would be of value in further studies, not only for the fabrication of complex shapes such as prosthetic ears, but also for other facial prostheses. PMID- 16220807 TI - The impact of tooth loss on general health related to quality of life among elderly Pomeranians: results from the study of health in Pomerania (SHIP-O). AB - PURPOSE: It is important to know whether tooth loss has an impact on an individual's quality of life, since tooth loss is common. The aim of this study was to determine whether oral status is associated with general health and related to quality of life. MATERIALS AND METHODS: Data of 1,406 subjects aged 60 to 79 years were taken from the Study of Health in Pomerania (SHIP). Sociodemographic (age, sex, and education level) and medical information (including the most common diseases in Germany) were gathered through an interview, and income data were obtained from a self-administered questionnaire. The prosthetic status in the maxilla and mandible was classified into complete denture or removable partial denture or with > or = 10 natural teeth including teeth replaced with fixed prosthodontics (> or = 10T) or with < or = 9 natural teeth including fixed prosthodontics (< or = 9T). The health-related quality of life was measured using the Short Form (SF-12) questionnaire. Multiple linear logistic regression analyses were used to identify the nonstandardized beta coefficient using physical and psychologic indices from the SF-12 as dependent variables and sociodemographic information, prosthetic status, and disease state as independent variables. RESULTS: Prosthetic status is related to the physical scale of the SF-12. Additionally, we found that < or = 9T had a significant effect on the physical index of general health-related quality of life. CONCLUSION: Reduced dentition without replacement of missing teeth by removable or fixed prosthodontics reduces the physical index of quality of life to the same extent as cancer or renal diseases. PMID- 16220808 TI - Effect of original water content in acrylic resin on processing shrinkage. AB - PURPOSE: The aim of this study was to determine the original water content of a supplied acrylic resin powder and a monomer and of dry heat-processed acrylic bars. The effect of the original water content of acrylic resin on processing shrinkage was also investigated. MATERIALS AND METHODS: Ten bar specimens were fabricated using dried and as-supplied (control) acrylic resins. The resins were polymerized and cooled, then weighed and measured to determine the amount of shrinkage. The initial water content of the specimens was determined by thorough drying, and results were compared with the Student ttest. RESULTS: The initial water content and processing shrinkage of the dried acrylic resin bars were both significantly lower (P < .0001) than those of the as-supplied acrylic resin bars. CONCLUSION: The processing shrinkage of acrylic resin made from dried constituents was significantly less than that of resin made from products as supplied by the manufacturer. However, it is not known if this change is of clinical significance. PMID- 16220809 TI - Clinical effect of different shade guide systems on the tooth shades of ceramic veneered restorations. AB - PURPOSE: The objective of this study was to investigate whether a systematically arranged shade guide system (Vita 3D-Master) allows clinicians to achieve a better shade match of a restoration, as compared to a conventional shade guide with a design based on empirical values (Vita Classical). MATERIALS AND METHODS: Fifty-nine restorations in 42 patients being treated by student clinicians were assessed. Using 1 of the 2 shade systems assigned randomly, each student independently determined the tooth shade. With the aid of a visual rating scale, the accuracy of the shade match of the finished restoration was assessed. RESULTS: All restorations whose shades had been determined with the 3D-Master could be placed without any further shade corrections. In contrast, almost 17% of restorations determined with the conventional system required subsequent shade modifications. The match of the shades selected with the 3D-Master was judged significantly better by the clinicians. CONCLUSION: Within the limitations of the study, clinicians with less clinical experience who use a system that guides them through the shade-taking procedure in a relatively systematic manner will be more successful in selecting the correct tooth shade and in avoiding shade corrections. Clinical assessment of the restoration shades showed significant differences between the shade guide with a systematic design and that based on empirical values. PMID- 16220810 TI - Peri-implant bone loss as a function of tooth-implant distance. AB - PURPOSE: In a retrospective study, the radiographs of 39 patients with Applegate Kennedy Class I or II in the posterior mandible who had been treated with screw anchored fixed partial dentures supported by IMZ implants and natural teeth were examined for the presence of radiologically detectable peri-implant bone loss. Furthermore, the results were correlated with a mathematical model. MATERIALS AND METHODS: The radiographs of the implants were digitized, and the areas of bone atrophy mesial and distal to the implants were determined semi-automatically. The data obtained were correlated with the distance between the implant and the abutment tooth. The connection between the tooth-supported crown and the implant supported denture was made with a vertical screw-lock precision attachment. In a mathematical analysis it was assumed that the fixed partial prosthesis was a rigid beam with 3 elastically embedded supports. RESULTS: The mean distance between the tooth and the first implant was 11.02 mm (SD: 4.24), and between the tooth and the second implant was 20.25 mm (SD: 5.16). Peri-implant bone loss significantly followed a rational function (mesial implant: P = .03, distal implant: P = .02), meaning that, as the tooth-implant distance increased, the area of atrophy became rapidly larger and then diminished gradually. Distances of 8 to 14 mm between the tooth and the first implant and of 17 to 21 mm between the tooth and the second implant were associated with a more pronounced bone loss. These results were also confirmed mathematically. CONCLUSION: A tooth-implant distance of 8 to 14 mm for the first implant and 17 to 21 mm for the second implant should be avoided for implant placement if prosthetic rehabilitation is planned using a fixed partial denture supported by a premolar and 2 IMZ implants in the mandible. Although this investigation was done on IMZ implants only, the results were confirmed by a mathematical model, which indicated that the observed bone loss may be the same in other types of implants placed in the same positions. PMID- 16220811 TI - Intensity of quartz-tungsten-halogen light polymerization units used in dental offices in Brazil. AB - PURPOSE: To determine the intensity of quartz-tungsten-halogen light polymerization units in dental offices in El Salvador, Brazil. MATERIALS AND METHODS: A team of researchers visited 100 offices in 2003. The team measured light intensity with a radiometer. The age and make of the units and the availability of light meters were recorded. RESULTS: One hundred twenty quartz tungsten-halogen units were tested. Intensity ranged from 10 to 1,000 mW/cm2, with mean of 255.8 mW/cm2. Almost half (48%) of the units had values equal to or less than 200 mW/cm2. Their age ranged from 1 to 21 years. None of the offices had light meters. CONCLUSIONS: Clinicians should periodically monitor light intensity and carry out maintenance on their light polymerization units. PMID- 16220812 TI - Use of shadow-speckle correlation method for 3D tooth model reconstruction. AB - This study presents a method of using shadow-speckle correlation to reconstruct a 3-dimensional (3D) tooth model. Compared with other methods based on laser or optical geometric measurements, the shadow-speckle method overcomes their limitations by using a digital image correlation to reconstruct a 3D tooth model. Using plaster models 4 times the normal tooth size showed that the accuracy of the reconstructed model was estimated at roughly 30 microm, which potentially could be used in direct intraoral applications. PMID- 16220813 TI - The reliability of digital cameras for color selection. AB - PURPOSE: This study evaluated the validity of the color selection results obtained from digital cameras. MATERIALS AND METHODS: Four observers matched the colors of 30 extracted maxillary incisors under daylight conditions. Two different digital cameras, with 1.5- and 5.0-megapixel resolutions, were used to select the color of the standardized circular area with 3 reference points on the same teeth; a spectrophotometer was also used, and the results were compared with the human and camera observations. RESULTS: The highest significant agreement (60% agreement with the spectrophotometric values) was found with the 5.0 megapixel camera. CONCLUSION: It appears that the resolution capacity of a digital camera can increase the reliability of color selection. PMID- 16220814 TI - ACHE's collaboration pays offs. Why relationships in today's environment help organizations succeed. PMID- 16220815 TI - The transformational power of IT: Experience from patient safety leaders. AB - Late last year, David J. Brailer, M.D., Ph.D., National Health Information Technology Coordinator for the Department of Health and Human Services, visited Evanston (IL) Northwestern Hospital (ENH) to observe its electronic medical record system. ENH is the first in the nation to put such a system into an acute care setting, including all three of its hospitals and its 70 medical offices and ambulatory sites. "It's been truly transformational for our processes, our delivery of care, our communications, and our way of doing business," says Mark R. Neaman, FACHE, ENH's president and chief executive officer. What's more, ENH has demonstrated at least a 20 percent reduction in medication errors, and the remaining errors have shifted from wrong medication and wrong patient to errors of timing, which are almost always less severe. PMID- 16220816 TI - Emerging benefits of integrated IT systems. AB - Technological innovation in healthcare is occurring at a rapid pace both in terms of clinical systems and systems that affect the overall management of an organization's operations. Electronic health records are currently in the spotlight for the Bush administration as the federal government is in the beginning phases of cultivating a national health information network. At the same time, chief executive officers and chief information officers around the country are focused on improving all of their IT operations right now. PMID- 16220817 TI - Facing change in an organization: how to chart your way through the chaos. PMID- 16220818 TI - Promoting advance directives. The administrators' role in encouraging advance directives. PMID- 16220819 TI - Tracking complaints. Prevent future service problems by identifying their source. PMID- 16220820 TI - Cover the Uninsured Week 2005. Healthcare organizations get involved in the community. PMID- 16220821 TI - Post-acute care payment policies. Why Medicare is looking to make care easier for patients. PMID- 16220822 TI - Getting a running start. A primer on successful job transition. PMID- 16220823 TI - Board meeting evaluation. Improve board performance through continuous inquiry. PMID- 16220824 TI - Going wireless. Hospitals combine current applications and future flexibility. PMID- 16220825 TI - Involving medical staff in marketing. Linking marketing and strategy yields results. PMID- 16220827 TI - Networking. Avoid crossing the line from persistence to annoyance. PMID- 16220828 TI - Prognostic value of GLUT-1 expression in ovarian surface epithelial tumors: a morphometric study. AB - OBJECTIVE: To investigate the reported increase in the expression of the glucose transporter GLUT-1 in borderline and malignant ovarian epithelial tumors and its relationship to prognosis. STUDY DESIGN: In this study, areas in which immunohistochemical membranous staining with GLUT-1 were most evident were selected, and the proportions of GLUT-1 expression in 46 benign, 11 borderline and 42 malignant cases of ovarian epithelial tumors were determined quantitatively with a computer and Zeiss Vision KS 400 3.0 (Gottingen, Germany) for Windows (Microsoft, Redmond, Washington, U.S.A.) image analysis. RESULTS: GLUT-1 expression was determined in all borderline tumors (11 of 11) and in 97.6% of malignant tumors (41 of 42). No GLUT-1 expression was observed in benign tumors. The intensity of GLUT-1 staining was lower in borderline tumors than in malignant cases. This was statistically significant (p = 0.005). As differentiation in malignant tumors increased, proportions of GLUT-1 expression showed a relative increase, but this difference was not statistically significant (p = 0.68). CONCLUSION: When GLUT-1 expression in borderline and malignant ovarian epithelial tumors was analyzed against prognosis, no statistically significant difference was identified. Assessment of GLUT-1 expression using the image analysis program was more reliable, with higher reproducibility than in previous studies. PMID- 16220826 TI - Recruitment and retention. Hiring the right people. PMID- 16220829 TI - Sampling of sentinel lymph nodes: a simulation study. AB - OBJECTIVE: To analyze the effects of various slicing schemes on the detection of metastases in lymph nodes. STUDY DESIGN: Use of an advanced computer simulation tool. RESULTS: Bisectioning along the longitudinal axis is an inadequate approach. Slicing ellipsoid lymph nodes along their longitudinal axis also results in a lower rate of detecting metastases since metastatic deposits have a predilection to localize subcapsularly. CONCLUSION: Ellipsoid lymph nodes must be sliced perpendicular to the longest axis to increase the rate of detecting metastases. PMID- 16220830 TI - Karyometry of infiltrating breast lesions. AB - OBJECTIVE: To characterize nuclei from well-differentiated, moderately differentiated and poorly differentiated lesions of invasive breast cancer by karyometry and to test the hypothesis that these diagnostic categories form homogeneous sets. STUDY DESIGN: Histopathologic sections from 6 cases of well differentiated, 11 cases of moderately differentiated and 17 cases of poorly differentiated ductal carcinomas were digitally recorded. From each case 100 nuclei were segmented and analyzed by karyometry. A discriminant analysis was performed, and nuclear and lesion signatures were computed. The nonsupervised learning algorithm P-index was applied. A progression curve per diagnostic category based on mean nuclear abnormality and a discriminant function score was derived. RESULTS: The well-differentiated lesions formed a homogeneous set, but both the moderately and poorly differentiated lesions showed 2 significantly different subpopulations with nuclei of substantially different nuclear abnormality and progression. CONCLUSION: The visual histopathologic diagnostic assessment of these lesions was based on an evaluation of both tissue architectural criteria and nuclear criteria. Here, only the pattern of nuclear chromatin was evaluated. Cases belonging to the same diagnostic category as assessed by their differentiation may be further characterized by the extent to which the nuclei deviate from normal. There was substantial case-to-case heterogeneity in these invasive lesions. PMID- 16220831 TI - Expression of PTEN in ovarian epithelial tumors and its relation to tumor behavior and growth. AB - OBJECTIVE: To evaluate the expression of tumor suppressor gene phosphatase and tensin homologue on chromosome 10 (PTEN) in ovarian epithelial tumors and its correlation with tumor growth and clinicopathologic features in ovarian adenocarcinomas. STUDY DESIGN: Immunohistochemical staining with anti-PTEN antibody was performed in 54 adenocarcinomas and 23 borderline tumors of the ovary. The apoptotic cells were visualized by terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling, and proliferative cells were visualized by staining with Ki-67 antibody. RESULTS: Reduced PTEN expression was significantly higher among the adenocarcinomas than the borderline tumors (p < 0.001). Reduced PTEN expression in adenocarcinomas did not correlate with International Federation of Obstetrics and Gynecology (FIGO) stage. The Ki-67 index (KI) and apoptotic index were significantly higher in adenocarcinomas as compared with borderline tumors (p < 0.001). Tumors with reduced PTEN expression in ovarian adenocarcinomas had a significantly higher KI than those with normal PTEN expression (p < 0.01). By univariate analysis, FIGO stage and histologic type correlated with survival. However, FIGO stage was the only independent prognostic factor by multivariate analysis. CONCLUSION: Our results suggest that alteration of the PTEN gene may be associated with malignant transformation of ovarian epithelial tumors. The PTEN gene seems to be a negative regulator of cell proliferation in ovarian adenocarcinomas. PMID- 16220832 TI - Prostate cancer outcome: epidemiology and biostatistics. AB - Substantial gaps exist in our ability to accurately predict prognosis, and these gaps limit our understanding of the complex mechanisms that contribute to the greatest cancer epidemic of our time, prostate cancer. This review addresses contemporary epidemiologic and biostatistical issues in prostate cancer. It covers the science of outcome prediction and biomarker evaluation, recognition of the need to combine biomarkers to improve the accuracy of our outcome estimates and an analysis of current outcome assessment methods, including the TNM staging system and multivariate regression models. The simplicity and intuitive ease of the current TNM staging system must be balanced against its serious limitations in predictive accuracy and its loss of clinical utility. Statistical regression methods are required as we move to the new era of personalized medicine. We must implement statistical approaches that integrate the new molecular biomarkers with existing prognostic biomarkers to accurately predict which patients require treatment and to determine the optimal therapy. PMID- 16220833 TI - Immunohistochemical study of neuroendocrine differentiation in primary glandular lesions and tumors of the urinary bladder. AB - OBJECTIVE: Neuroendocrine (NE) cells are uncommon in primary adenocarcinoma (AC) and other glandular lesions of the bladder, with no recent study series concerning its significance in differential diagnosis, prognosis or biologic significance. STUDY DESIGN: Sixteen primary bladder AC (enteric-type [n = 71, mucinous [n = 6] and not otherwise specified [NOS] [n = 31), 4 cases of urothelial carcinoma with glandular differentiation, 20 cases of glandular cystitis and 3 urachal remnants with intestinal metaplasia constituted the study series. In addition, 20 specimens of normal-looking urothelium, 15 conventional urothelial carcinomas and 5 small cell carcinoma (SCC) cases were included for comparison. NE differentiation included detection of chromogranin A, neuron specific enolase (NSE) and synaptophysin by immunohistochemistry. The statistical analysis included the chi2 or Fisher exact test. RESULTS: Chromogranin A-positive cells were present in 60% (11 of 16) of primary AC, all of enteric or mucinous type, but not in any of the 3 NOS-type AC investigated. NE differentiation in bladder AC subtypes resulted in highly significant differences between enteric or mucinous vs. NOS type (p = 0.0023). NE differentiation was also different in urachal vs. nonurachal AC (p = 0.020) and primary bladder AC vs. conventional invasive urothelial carcinoma (p < 0.001). Synaptophysin-positive cells were seen in 2 (12.5%) of the 16 primary AC cases, and NSE was negative in the 16 primary bladder AC. All urachal remnants and 70% of glandular cystitis examples had chromogranin A-immunoreactive cells. One of 4 urothelial carcinomas with glandular differentiation had chromogranin A-immunoreactive cells, but this was not significant when compared with primary AC (p = 0.1). Normal-looking bladder urothelium and conventional urothelial carcinoma specimens had no chromogranin A immunoreactive cells. The 5 SCC cases investigated were positive for chromogranin A. No correlation was found between NE differentiation and outcome of primary bladder AC or urothelial carcinoma with glandular differentiation. CONCLUSION: Primary bladder AC, cystitis glandularis and urachal remnants with intestinal metaplasia showed variable degrees of NE differentiation, with no apparent clinical correlation or prognostic significance. However, the absence of NE differentiation in NOS-type primary bladder AC may help in better defining this uncommon subtype of primary bladder AC. PMID- 16220834 TI - A simple procedure for simulating samples of tissue using Voronoi diagrams. AB - OBJECTIVE: To describe a simple and quick procedure for modeling samples of tissue with Voronoi diagrams. STUDY DESIGN: Instead of calculating the centers of the so-called Dirichlet domains (i.e., the polygonal areas occupied by individual cells), the centroid of such areas is used to generate Voronoi diagrams. The coordinates of the centroids are calculated by simply averaging the coordinates of the points of the cell contours; that is much simpler and faster than any geometric procedure for locating the Dirichlet centers. Using the centroids as centers, circles are allowed to grow until no space on the surface is available. With this procedure it is easy to control the rate of growth of individual cells or groups of cells according to any rule or rules. It is also possible to simulate the effects of removing > or = 1 cells from the sample. CONCLUSION: The procedure was successfully applied to modeling some of the changes that can occur in a real sample of human corneal endothelium. The procedure is simple, completely automated, efficient and flexible and can be easily implemented on a personal computer. It can be used to test growth or communication strategies among cells. PMID- 16220835 TI - Morphometry of pseudoepitheliomatous hyperplasia: objective comparison to normal and dysplastic oral mucosae. AB - OBJECTIVE: To compare the architectural and morphometric features of pseudoepitheliomatous hyperplasia (PEH) associated with oral granular cell tumors (GCT), normal oral mucosa and oral epithelial dysplasia. STUDY DESIGN: Quantitative comparisons between the diagnostic entities were carried out at the tissue level by estimating the fractal complexity of the epithelial connective tissue interface and at the cellular level by analyzing the morphometric features of algorithmically segmented epithelial cell areas. RESULTS: Casewise multivariate analysis showed that the fractal properties produced a correct discrimination rate of 96.4% between PEH and normal mucosa. Cellular parameters gave a 100% correct discrimination rate between PEH and mild dysplasia. Combining the fractal and cellular properties also showed 100% discrimination between PEH and normal mucosa and between PEH and mild dysplasia. CONCLUSION: The results show that PEH associated with GCT displays quantifiable morphometric features that make it differentiable from normal oral mucosa and oral epithelial dysplasia. PMID- 16220837 TI - Using unstructured interviews with participants who have schizophrenia. AB - Unstructured interviews offer a potential source of rich data in nursing research. In this paper, Terence McCann and Eileen Clark examine the challenges presented when unstructured interviews are used with participants who have schizophrenia. They consider unstructured interviewing in the context of an Australian study of how community mental health nurses promote wellness with clients who are experiencing an early episode of schizophrenia. PMID- 16220836 TI - Blood bank information systems. Attention to errors linked to usefulness of wares. PMID- 16220838 TI - A reflection on the use of semi-structured interviews. AB - Based on the her experiences of completing a doctoral study in which semi structured interviews featured as the primary data collection method, Christine Dearnley offers a reflective insight into using semi-structured interviews as a method of data collection. The processes of reflection in, and on, the interview process are explored, and some of the ethical dilemmas that emerged during the study are reflected on. The practicalities of conducting semi-structured interviews are considered with a view to sharing new understandings of the process and its management. PMID- 16220839 TI - Part II. rigour in qualitative research: complexities and solutions. AB - Anthony G Tuckett outlines the strategies and operational techniques he used to attain rigour in a qualitative research study through relying on Guba and Lincoln's trustworthiness criterion. Research strategies such as use of personal journals, audio recording and transcript auditing, and operational techniques including triangulation strategies and peer review, are examined. PMID- 16220840 TI - The narrative research trail: values of ambiguity and relationships. AB - Engle Angela Chan sets out the experiential function of ambiguity in nursing research through a narrative study of Canadian college nurse educators. Through the methods of conversation, journal writing and participation observation, the author, who was also one of the participants, explored with her colleagues the impact of nursing education and healthcare changes on their teaching lives. The article examines how researcher-participant relationships illuminate research outcomes. PMID- 16220841 TI - Creating user-friendly databases with Microsoft Access. AB - Data entry can be tedious and is fraught with potential for errors that affect study findings. Researchers can minimise entry errors and streamline data entry by using some of the popular software packages on the market. Joanne Kraenzle Schneider and colleagues describe one way to create a user-friendly database that minimises entry errors by using Microsoft (MS) Access. PMID- 16220842 TI - The challenge of recruitment. AB - Paul Affleck reflects on the challenges posed to researchers in recruiting patients to epidemiological studies. Computerised patient information sources, he argues, offer a valid and accessible means of assisting recruiting processes, with a cancer registry being cited as an example of an appropriate database that can be searched quickly and efficiently to identify possible participants. PMID- 16220843 TI - 'Catch it, encode it and measure it'. AB - Donna Mead, Head of School, School of Care Sciences, University of Glamorgan, explains how her early experiences as a ward sister gave her the drive and determination to seek research-based solutions to some of the conditions that she encountered. PMID- 16220844 TI - Arts medicine: an interdisciplinary paradigm. PMID- 16220845 TI - Otitis media with effusion in a patient who had previously undergone a stapedectomy. PMID- 16220846 TI - Asymmetry of the middle turbinates and ethmoid sinuses. PMID- 16220847 TI - Dense vocal fold scar. PMID- 16220848 TI - Myeloid sarcoma. PMID- 16220849 TI - Carrier ID numbers. PMID- 16220851 TI - Dysphagia and reflux caused by a giant hiatal hernia. PMID- 16220850 TI - Sialadenoma papilliferum involving the nasopharynx. PMID- 16220852 TI - Vestibular ENG findings in a 46-year-old woman with dizziness and an autoimmune disease. PMID- 16220853 TI - Potential applications of the da Vinci minimally invasive surgical robotic system in otolaryngology. AB - Anatomic constraints and instrumentation design characteristics have limited the exploitation of endoscopic surgery in otolaryngology. The move toward less invasive and less morbid procedures has paved the way for the development and application of robotic and computer-assisted systems in surgery. Surgical robotics allows for the use of new instrumentation in our field. We review the operative advantages, limitations, and possible surgical applications of the da Vinci Surgical System in otolaryngology. In the laboratory setting, we explored the setup and use of the da Vinci system in porcine and cadaveric head and neck airway models; the setup was configuredfor optimal airway surgery. Endoscopic cautery, manipulation, and suturing of supraglottic tissues were performed in both the porcine and cadaveric models. We found that the da Vinci system provided the advantages of the lower morbidity associated with endoscopic surgery, more freedom of movement, and three-dimensional open surgical viewing. We also observed that the system has several limitations to use in otolaryngology. PMID- 16220854 TI - Tuberculous otitis media: report of 2 cases on Long Island, N.Y., and a review of all cases reported in the United States from 1990 through 2003. AB - We report 2 cases of tuberculous otitis media that were diagnosed at Stony Brook University Hospital in New York since 1999. Both patients were women, aged 30 and 31 years. One patient had grown up in Russia, the other was a native-born American who had never left the East Coast region of the United States. Both patients had been symptomaticfor many months; one complainedof chronic otorrhea, and the other reported otorrhea, hearing loss, and discomfort. Neither patient responded to medical management, and both ultimately underwent surgery. One was diagnosed after surgical pathology revealed acid-fast bacilli on frozen-section analysis. In the other, pathology revealed chronic inflammation and granulomata, butstains were negative and her diagnosis was delayed for almost 2 years. We also review 9 other cases of tuberculous otitis media in the United States that have been reported in the literature since 1990. Our review suggests that the number of cases is rising in areas where tuberculosis is most common--that is, in major U.S. cities. Although 3 of these 9 cases occurred as reactivation disease in immigrants, most might have occurred as a result of local transmission. Clinicians should maintain a high degree of suspicion for tuberculosis in patients with chronic otitis symptoms, particularly those who are at higher risk of exposure to tuberculosis. PMID- 16220855 TI - Cochlear implantation in severe traumatic brain-injury and paranoid schizophrenia: two case studies. AB - In view of the demanding instruction and training requirements associated with cochlear implantation;,patients with cognitive impairments have typically been considered to be poor candidates for this procedure. This presumption persists in part because experience and research in this area are limited. We describe our experience with implanting cochlear devices in 2 patients who had significant psychological impairments; 1 patient had experienced a severe brain injury, and the other had paranoid schizophrenia. Nevertheless, both patients were able to follow the implant training program, and they experienced different degrees of improvement in their hearing. We discuss our preoperative evaluations of these patients and the key factors that led to our decision to proceed with surgery. PMID- 16220856 TI - Bilateral massive conchae bullosa mimicking intranasal tumors. AB - We describe a case of bilateral massive conchae bullosa in a 76-year-old woman. She presented with a 2-year history of nasal obstruction and frontal headache. In light of these and other findings on anterior rhinoscopic and endoscopic examinations, we initially suspected nasal tumors. However, after a prebiopsy evaluation by computed tomography, we diagnosed bilateral massive conchae bullosa that did not impair sinus ventilation. Endoscopic surgery was performed, and the patient's symptoms abated. PMID- 16220857 TI - Total necrosis of the intranasal structures and soft palate as a result of nasal inhalation of crushed OxyContin. AB - Nasal inhalation of substances such as cocaine has long been linked to septal necrosis and more recently to palatal perforation. This report describes the case of a 37-year-old man who was addicted to crushed OxyContin (sustained-release oxycodone HCl) tablets and who presented with total necrosis of the septum, sinuses, and soft palate. To the best of the author's knowledge, this is the first reported case of nasal and palatal necrosis linked to nasal inhalation of crushed OxyContin, which is a relatively new form of drug abuse. The pathophysiology of drug-induced sinonasal disease and a review of the literature are also presented. PMID- 16220858 TI - Preservation of the superficial lobe for deep-lobe parotid tumors: a better aesthetic outcome. AB - Deep-lobe parotid tumors are relatively uncommon. Most of these tumors present as external masses. They can also present in the oral cavity or oropharynx. Magnetic resonance imaging and ultrasound-guided fine-needle aspiration for biopsy and cytology have made it possible to establish a definitive diagnosis and identify the exact location of the tumor in almost all cases before surgery. Traditionally, deep-lobe tumors have been managed by a formal superficial parotidectomy and identification and preservation of the facial nerve, followed by removal of the deep lobe that contains the tumor Superficialparotidectomy is associated in most cases with periauricular depression secondary to a loss of volume, leading to variable aesthetic deformities. A complete parotidectomy is more likely to be associated with a larger aesthetic deficit secondary to a greater loss of tissue volume. The incidence of gustatory sweating is high after superficial parotidectomy, particularly in the early postoperative period. We hypothesize that if the superficial lobe is preserved, there is less likelihood of gustatory sweating because of the interposition of tissue between the skin and the cut ends of the secretomotor fibers. Approximately 80% of parotid tissue volume is made up of the superficial lobe, and therefore preservation of the superficial lobe should be associated with less postparotidectomy depression. Therefore, we decided to preserve the superficial lobe of the gland for deep-lobe tumors. Nine patients underwent deep-lobe parotidectomy with preservation of the superficial lobe over a 6-year period. Patients were studied prospectively with regard to technical difficulty, complications, and cosmetic outcome. Follow-up ranged from 12 months to 6 years. We did not experience any undue technical difficulty, and there were no cases of facial weakness. One patient developed gustatory sweating, which almost completely resolved over a 2-year period. There were no cases of post parotidectomy depression, and both patients and surgeons were satisfied with the cosmetic appearance. We present our technique and experience. PMID- 16220859 TI - An evaluation of the Australian Sentinel Practice Research Network (ASPREN) surveillance for influenza-like illness. AB - The Australian Sentinel Practice Research Network (ASPREN) is a national network of general practitioners (GPs) who collect and report data on selected conditions, including influenza-like illness (ILI). The Australian Government Department of Health and Ageing initiated an evaluation of ASPREN, aiming to assess its potential to contribute to surveillance of emerging infectious diseases including pandemic influenza. System attributes and utility for decision making were elucidated from stakeholder surveys. ASPREN ILI data for 2002 to 2004 were compared with ILI data from South Australia and New South Wales. In 2004, 50 GPs participated in the ASPREN surveillance, with proportionately more in New South Wales (30%) and South Australia (30%) than in other states. The majority (78%) of GPs were in metropolitan practices. Compliance with the manual data collection system was not optimal, nor consistent by state. ASPREN ILI data compared favourably with that of other surveillance systems. No formal structures were in place by which to assess data trends, provide alerts or initiate public health action. To maximise the contribution to biosecurity surveillance, ASPREN would require targeted GP recruitment to achieve geographic representativeness; exploration of alternative technologies for data collection and reporting; provision of committed resources adequate for system operation; and negotiation with state-based public health reference laboratories to provide laboratory support. The main potential of ASPREN is to permit rapid dissemination of a syndromic case definition and acquisition of nationwide community level clinical presentation data. These evaluation findings will be used to inform redevelopment of ASPREN as part of the Biosecurity Surveillance System project. PMID- 16220860 TI - Annual report: surveillance of adverse events following immunisation in Australia, 2004. AB - This report summarises Australian passive surveillance data on adverse events following immunisation (AEFI) for 2004 and describes reporting trends over the five years, 2000 to 2004. AEFIs are notified to the Adverse Drug Reactions Advisory Committee by state and territory health departments, hospitals, doctors and other health providers, vaccine manufactures, and the public. There were 975 AEFI records for vaccines received in 2004. This is an annual AEFI reporting rate of 4.8 per 100,000 population, the lowest since 2000, and a 33 per cent decrease compared with 2003 (1,460 records; 7.1 AEFI records per 100,000 population). Dose based AEFI reporting rates in 2004 were 1.8 per 100,000 doses of influenza vaccine for adults aged > or = 18 years and 11.8 per 100,000 doses of scheduled vaccines for children aged < 7 years. The majority of records described non serious events while nine per cent (n = 88) described AEFIs defined as 'serious'. There were no reports of death related to immunisation. The most frequently reported individual AEFI was injection site reaction in children following a fifth dose of an acellular pertussis-containing vaccine (67 reports per 100,000 doses). The marked reduction in the AEFI reporting rate in 2004 coincided with the removal of the fourth dose of acellular pertussis vaccine, due at 18 months of age, from the vaccination schedule in September 2003 and fewer people receiving meningococcal C vaccine through the national catch-up vaccination program for those aged 1-19 years in 2004, compared with 2003. The consistently low reporting rate of serious AEFIs demonstrates the high level of safety of vaccines in Australia. PMID- 16220861 TI - Annual report of the Australian National Poliovirus Reference Laboratory, 2004. AB - The Australian National Poliovirus Reference Laboratory at the Victorian Infectious Diseases Reference Laboratory is the World Health Organization designated laboratory for the isolation and testing of poliovirus from clinical specimens within Australia, the Pacific Island countries and Brunei Darussalam. Surveillance for acute flaccid paralysis (AFP) within Australia, the main clinical manifestation of poliomyelitis, is also coordinated at the Victorian Infectious Diseases Reference Laboratory in conjunction with the Australian Paediatric Surveillance Unit. The annual non-polio acute flaccid paralysis rate after classification of cases by the Polio Expert Committee was 1.0 per 100,000 population, reaching the expected World Health Organization annual target for a non-polio endemic country. During 2004, 64 specimens from 30 AFP cases were referred to the National Polio Reference Laboratory. A mixture of poliovirus types 1 and 2 was isolated from an infant with AFP from New South Wales. Both isolates tested as Sabin-like and the case was subsequently classified as infant botulism by the Polio Expert Committee. The laboratory isolated adenoviruses from seven AFP cases. A coxsackievirus B5 and an echovirus 18 were identified from a further two AFP cases. During 2004, 1,266 cases of poliomyelitis due to wild poliovirus were reported world-wide. Many of these resulted from wild poliovirus importations, which continued in 2005, including to Indonesia. This highlights the need for maintaining high poliovirus vaccination coverage to prevent the transmission of poliovirus and high quality AFPand laboratory surveillance for the detection of poliomyelitis due to an imported wild poliovirus. PMID- 16220862 TI - Creutzfeldt-Jakob disease: Australian surveillance update to 31 December 2004. AB - The Australian National Creutzfeldt-Jakob Disease Registry (ANCJDR) was established in October 1993 after the identification of probable iatrogenic CJD in recipients of human pituitary hormones. Since this time and with the recommendations of the Allars inquiry into CJD in Australia, the registry has performed surveillance of CJD in Australia with retrospective ascertainment to 1970 and ongoing prospective ascertainment of all human prion diseases or transmissible spongiform encephalopathies (TSEs). Prion diseases include CJD, Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia and Kuru. This brief summary presents the epidemiological findings of the ANCJDR based on data from 1970 to 31 December, 2004. PMID- 16220863 TI - A comparison of a rapid test for influenza with laboratory-based diagnosis in a paediatric population. AB - The rapid and accurate detection of influenza A and B in a hospital setting is useful to confirm infection, exclude other diseases and assist in the management of patient illness including the possible use of specific antiviral therapy. We evaluated the use of the Directigen Flu A+B in a paediatric hospital laboratory in comparison with the established diagnostic tests direct immunofluorescence, viral culture and reverse transcriptase-polymerase chain reaction. A total of 193 respiratory specimens were examined and the Directigen test detected positive samples with an 80.8 per cent sensitivity and a specificity of 100 per cent. This study confirms other paediatric studies which have found the Directigen Flu A+B to be less sensitive than traditional laboratory tests but nevertheless to have a potential role in patient management especially when a positive result is obtained. PMID- 16220864 TI - SARS and biothreat preparedness--a survey of ACT general practitioners. AB - In late 2003 and early 2004 the ACT Division of General Practice and ACT Health conducted two concurrent surveys designed to identify knowledge, attitudes and practices of Australian Capital Territory (ACT) general practitioners around severe acute respiratory syndrome (SARS) and biothreat preparedness. One survey asked individual general practitioners (GPs) about how they gathered information about SARS in 2003, how they preferred to receive information, current practices, and how they perceived the threat of SARS and other infectious agents. The second survey asked practice principals how they organised their practice to respond to the SARS threat in 2003, any difficulties they had with implementing this response, use of SARS infection control guidelines and current policies. The response rate for the individual GP survey was 48 per cent (184/381) and the response rate for the practice organisation survey was 54 per cent (74/136). GPS used many sources of information on SARS during the 2003 outbreak. Facsimiles from the ACT Division of General Practice were the primary source (17%) and facsimile was the preferred method of receiving information in future outbreaks. The majority of GP respondents felt adequately informed about SARS during the 2003 outbreak, but many general practices did not follow the national guidelines on telephone screening of patients, warning signs and having infection control kits available. The majority of practices reported that they had policies or procedures in place to isolate potentially infectious patients from others in the waiting room. GPs rated an influenza pandemic as a threat to themselves and their patients much more highly than SARS or bioterrorism. Suggestions and comments on how ACT GPs could be better prepared to respond to future outbreaks included the need for timeliness of information, information delivery mechanisms, communication issues, education, the availability of guidelines and protocols, planning, role delineation, the use of response teams, provision of equipment, and vaccination. Planning for future infectious disease outbreak events in the Australian Capital Territory should incorporate general practitioners so that the plans reflect what is a feasible response in the general practice setting. PMID- 16220865 TI - Influenza and pneumococcal vaccine coverage among a random sample of hospitalised persons aged 65 years or more, Victoria. AB - This study was undertaken to assess the uptake of influenza and pneumococcal vaccination based on provider records of the hospitalised elderly, a group at high risk of influenza and pneumococcal disease. The study used a random sample of 3,204 admissions at two Victorian teaching hospitals for patients, aged 65 years or more who were discharged between 1 April 2000 and 31 March 2002. Information on whether the patient had received an influenza vaccination within the year prior to admission or pneumococcal vaccination within the previous five years was ascertained from the patient's nominated medical practitioner/vaccine provider. Vaccination records were obtained from providers for 82 per cent (2,804/2,934) of eligible subjects. Influenza vaccine coverage was 70.9 per cent (95% CI 68.9-72.9), pneumococcal coverage was 52.6 per cent (95% CI 50.4-54.8) and 46.6 per cent (95% CI 44.4-48.8) had received both vaccines. Coverage for each vaccine increased seven per cent over the two study years. For pneumococcal vaccination, there was a marked increase in 1998 coinciding with the introduction of Victoria's publicly funded program. Influenza and pneumococcal vaccine coverage in eligible hospitalised adults was similar to, but did not exceed, estimates in the general elderly population. Pneumococcal vaccination coverage reflected the availability of vaccine through Victoria's publicly funded program. A nationally funded pneumococcal vaccination program for the elderly, as announced recently, should improve coverage. However, these data highlight the need for greater awareness of pneumococcal vaccine among practitioners and for systematic recording of vaccination status, as many of these subjects will soon become eligible for revaccination. PMID- 16220866 TI - Estimates of chronic hepatitis B virus infection in the Northern Territory. PMID- 16220867 TI - The incidence of Ross River virus disease in South Australia, 1992 to 2003. AB - Ross River virus (RRV) disease is the most frequently notified arboviral disease in Australia, and the burden of this disease to Australian society is significant. We have studied the incidence of RRV disease between 1992 and 2003 in South Australia. Our findings suggest that the incidence of the disease in South Australia over the study period was relatively stable. There were four epidemics in the study period, with the majority of cases acquired from regions along the River Murray. There was some evidence of spread of the disease to regions in which activity of RRV had not been previously recognised, such as the Mid-North and the South-East. In terms of disease distribution amongst the population, it was found that the highest rates occurred in the 30-49 year age range. There was no significant difference in disease rates between males and females. In order to facilitate further research into RRV disease transmission, we recommend that the suspected region of acquisition be a mandatory component of the national notification dataset. PMID- 16220868 TI - Aedes (Stegomyia) albopictus--a dengue threat for southern Australia? AB - Aedes albopictus, the so-called 'Asian tiger mosquito,' which has invaded areas of the Pacific, the Americas, Africa and Europe, and been intercepted in various Australian seaports in recent years, has now become established on a number of Torres Strait islands in northern Queensland and threatens to invade mainland Australia. As well as being a significant pest with day-biting tendencies, Ae. albopictus is a vector of dengue viruses and is capable of transmitting a number of other arboviruses. The species colonises domestic and peri-domestic containers, and can establish in temperate areas with cold winters. According to predictions made using the CSIRO climate matching software CLIMEX, Ae. albopictus could become established elsewhere in Australia, including southern Australia, and lead to these areas becoming receptive to dengue infections-a condition that currently does not exist because the vector Aedes aegypti is confined to Queensland and no species in southern Australia is known to be capable of transmitting dengue. PMID- 16220869 TI - Genetic diversity of the dengue vector Aedes aegypti in Australia and implications for future surveillance and mainland incursion monitoring. AB - In February 2004, the discovery of an incursion of the dengue vector Aedes aegypti into the town of Tennant Creek in the Northern Territory caused concern for the Northern Territory health authorities who proceeded to implement a Commonwealth-funded eradication program. To determine the origin of the incursion, we performed a genetic analysis on Ae. aegypti from several Queensland and overseas localities. A comparison of DNA sequences from the mitochondrial cytochrome oxidase 1 gene indicated that the incursion was probably from Cairns or Camooweal. This genetic marker was also useful in identifying a separate Townsville haplotype population and another population on Thursday Island in the Torres Strait that was genetically divergent to the mainland populations. The possible use of this marker as a surveillance tool for identifying the origins of local and overseas incursions is discussed. PMID- 16220870 TI - Septicaemia secondary to Vibrio vulnificus cellulitis. AB - Vibrio vulnificus is a naturally occurring, salt-water bacteria found in estuarine and coastal waters worldwide. It prefers low salinity and warm water temperatures for optimum growth. Infection from Vibrio vulnificus is uncommon, although it has been reported from many locations (e.g. southern United States of America, Israel, Republic of Korea, Japan, Taiwan, Spain, Turkey). It can be serious and life threatening, causing septicaemia and wound infections. This paper reports a case of septicaemia secondary to Vibrio vulnificus cellulitis in an elderly woman. The infection was acquired after wading in a coastal lagoon with a pre-existing superficial leg wound. PMID- 16220871 TI - OzFoodNet: enhancing foodborne disease surveillance across Australia: quarterly report, April to June 2005. PMID- 16220872 TI - Meningococcal disease--probable transmission during an international flight. AB - Two cases of meningococcal disease were identified in passengers who travelled on the same international flight. Both cases were serogroup B with the same allelic profile. The public health action involved chemoprophylaxis for persons seated adjacent to, and in the rows in front and behind, each case. The most likely scenario is that transmission of N. meningitidis occurred on board a long distance flight, either from one case to the other or from an asymptomatic carrier to both cases. This scenario and the absence of reports of similar cases in the literature, indicate the risk to other passengers in this setting is low. This investigation reinforces the need for, and the distribution of, good national and international surveillance information to better inform public health decision making. PMID- 16220873 TI - A report from the Communicable Diseases Network Australia, 1 April to 30 June 2005. PMID- 16220874 TI - Communicable diseases surveillance. Highlights for 2nd quarter, 2005. PMID- 16220875 TI - Bioterrorism and mass casualty preparedness in hospitals: United States, 2003. AB - OBJECTIVES: This study examined the content of hospital terrorism preparedness emergency response plans; whether those plans had been updated since September 11, 2001; collaboration of hospitals with outside organizations; clinician training in the management of biological, chemical, explosive, and nuclear exposures; drills on the response plans; and equipment and bed capacity. METHODS: The National Hospital Ambulatory Medical Care Survey (NHAMCS) is an annual survey of a probability sample of approximately 500 non-Federal general and short-stay hospitals in the United States. A Bioterrorism and Mass Casualty Supplement was included in the 2003 survey and provided the data for this analysis. RESULTS: Almost all hospitals have plans for responding to natural disasters (97.3 percent). Most have plans for responding to chemical (85.5 percent), biological (84.8 percent), nuclear or radiological (77.2 percent), and explosive incidents (76.9 percent). About three-quarters of hospitals were integrated into community wide disaster plans (76.4 percent), and 75.9 percent specifically reported a cooperative planning process with other local health care facilities. Despite these plans, only 46.1 percent reported written memoranda of understanding with these facilities to accept inpatients during a declared disaster. Hospitals varied widely in their plans for re-arranging schedules and space in the event of a disaster. Training for hospital incident command and smallpox, anthrax, chemical, and radiological exposures was ahead of training for other infectious diseases. The percentage of hospitals training their staff in any exposure varied from 92.1 percent for nurses to 49.2 percent for medical residents. Drills for natural disasters occurred more often than those for chemical, biological, explosive, nuclear, and epidemic incidents. More hospitals staged drills for biological attacks than for severe epidemics. Despite explosions being the most common form of terrorism, drills for these were staged by only one-fifth of hospitals. Hospitals collaborated on drills most often with emergency medical services, fire departments, and law enforcement agencies. PMID- 16220876 TI - CMS makes room for DM as Medicare health support debuts. PMID- 16220877 TI - Shifting the focus to new parameters for a burgeoning senior population. AB - Consider new parameters when caring for an older population. Why? Because clinical practice guidelines often fail to take into account the needs of people with multiple chronic conditions. Further, while some experts make the case that quality in this population can best be defined in individual terms, they suggest that that there are some new benchmarks policy makers should consider when developing quality improvement initiatives. PMID- 16220878 TI - New diabetes prediction tool lets patients, providers take glimpse into the future. AB - A diabetes risk prediction tool for providers and patients. An innovative computer model that has essentially turned anatomical processes into mathematical equations is now being used to help patients and their physicians mitigate their risk for diabetes or its complications. This is being done through an easily accessible online tool that developers maintain is far more accurate than previous risk prediction tools--and it may well be just a hint of things to come. PMID- 16220879 TI - Looking ahead to when the patient becomes the customer. AB - Preparing for a customer-driven marketplace. The sands are shifting in the health care marketplace, and at least one Orlando, FL-based company is betting that in the not-too-distant future consumers will have much more control over their health care, and DM in particular. To respond, the company is designing programs that put a premium on personalization and take a much broader view of ROI. PMID- 16220886 TI - [Leg ulcers: peri-ulcerous dermatitis]. PMID- 16220887 TI - [Myopathies and care]. PMID- 16220888 TI - [Clinical approach in myopathies]. PMID- 16220889 TI - [From diagnosis of the disease until today, a father's experience. Interview by Ghislaine Trabacchi]. PMID- 16220890 TI - [Announcement of the myopathy diagnosis]. PMID- 16220891 TI - [Genetic counseling]. PMID- 16220892 TI - [From diagnosis of the disease until today, a mother's experience. Interview by Ghislaine Trabacci]. PMID- 16220893 TI - [Respiratory management of muscular dystrophies]. PMID- 16220894 TI - [Nutritional approach in neuromuscular diseases]. PMID- 16220895 TI - [A nursing presence for an autonomous life at home]. PMID- 16220896 TI - [Ergonomic management of patients with myopathies]. PMID- 16220897 TI - [Children with myopathies and caregivers, resolution in the life direction]. PMID- 16220899 TI - [Bibliography elements. Myopathies and care]. PMID- 16220898 TI - [We are in the era of therapies]. PMID- 16220900 TI - [1/4 Preventing violence in health care]. PMID- 16220902 TI - Sonochemistry and sonoluminescence of room-temperature ionic liquids. AB - The sonication of ionic organic liquids leads to decomposition of the liquids. Multibubble sonoluminescence spectra and headgas analysis reveal a variety of decomposition products from the sonolysis of N,N'-dialkylimidazolium ionic liquids. The decomposition is a result of acoustic cavitation, which generates localized hot spots from the implosive collapse of bubbles in the ionic liquids. Despite the negligible vapor pressure of the ionic liquids, reaction still occurs in a heated shell of the bubbles or from microdroplets thrown into the collapsing bubbles. PMID- 16220901 TI - [III--Relationship care at the family level. 4. Helping parents who are perturbed in the exercise of their role]. PMID- 16220903 TI - Highly flexible polyelectrolyte nanotubes. AB - A pressure-filter-template approach was employed to prepare polyelectrolyte nanotubes through layer-by-layer deposition in the alumina template. With the thicker wall, the ordered polymer nanotubes possess a high flexibility. The results demonstrate that the electrostatic interactions of polyelectrolytes play a key role in fabricating water-soluble charged polymer nanotubes. The structure of the polyelectrolyte nanotube was confirmed by SEM, TEM, and UV, respectively. PMID- 16220904 TI - Cation-pi binding of an alkali metal ion by pendant alpha,alpha-dimethylbenzyl groups within a dinuclear iron(III) structural unit. AB - We report here on the cation-pi binding of potassium ions by benzyl groups in a coordination complex. The results demonstrate the cation-binding power of the benzyl group and consequently the potential for aromatic groups to interact with alkali metal ions even in aqueous media. PMID- 16220905 TI - The mechanism of vesicle fusion as revealed by molecular dynamics simulations. AB - We describe molecular dynamics simulations elucidating the molecular details of the process of fusion for small lipid vesicles. The simulations are based on a coarse grained (CG) lipid model that accurately represents the lamellar state of a variety of phospholipids and enables us to observe intermediate stages during fusion at near atomic detail. Simulations were conducted on a variety of systems containing common phospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), lysoPC, and mixtures of the above. The fusion intermediates found are in general agreement with the stalk-pore mechanism. Transient pores sometimes form adjacent to the stalk, however, resulting in the mixing of lipids from the outer and inner monolayers. The speed of stalk formation and the opening of the fusion pore can be modulated by altering the lipid composition in qualitative agreement with experimental observations. PMID- 16220906 TI - Novel fluorogenic substrates for imaging beta-lactamase gene expression. AB - A new class of small nonfluorescent fluorogenic substrates becomes brightly fluorescent after beta-lactamase hydrolysis with up to 153-fold enhancement in the fluorescence intensity. Less than 500 fM of beta-lactamase in cell lysates can be readily detected, and beta-lactamase expression in living cells can be imaged with a red fluorescence derivative. These new fluorogenic substrates should find uses in clinical diagnostics and facilitate the applications of beta lactamase as a biosensor. PMID- 16220907 TI - Expanding organoboron chemistry: epoxidation of potassium organotrifluoroborates. AB - Most organoboron reagents are highly susceptible to oxidation by a variety of oxidants. In these processes, the valuable carbon-boron bond is cleaved. The organotrifluoroborates examined in this contribution appear resistant to many common oxidants. Consequently, thioether-substituted alkyltrifluoroborates can be oxidized to the corresponding sulfones, while retaining the trifluoroborate moiety. 1-Trifluoroboratoalkenes are oxidized by dioxirane, providing air-stable, crystalline oxiranyltrifluoroborates. These interesting materials are structurally analogous to oxiranyl anions, unstable intermediates that would otherwise be of tremendous synthetic utility. Finally, the first Suzuki-Miyaura coupling of an epoxytrifluoroborate has been accomplished. The chemistry described begins to expand organoboron chemistry in significant new directions. PMID- 16220908 TI - Stopped-flow Fourier transform infrared spectroscopy of nitromethane oxidation by the diiron(IV) intermediate of methane monooxygenase. AB - The hydroxylase component (MMOH) of soluble methane monooxygenase from Methylococcus capsulatus (Bath) was reduced to the diiron(II) form and then allowed to react with dioxygen to generate the diiron(IV) intermediate Q in the first phase of a double-mixing stopped-flow experiment. CD3NO2 was then introduced in the second phase of the experiment, which was carried out in D2O at 25 degrees C. The kinetics of the reaction of the substrate with Q were monitored by stopped-flow Fourier transform infrared spectroscopy, observing the disappearance of the asymmetric NO2 bending vibration at 1548 cm-1. The data were fit to a single-exponential function, which yielded a kobs of 0.45 +/- 0.07 s-1. This result is in quantitative agreement with a kobs of 0.39 +/- 0.01 s-1 obtained by observing the disappearance of Q by double-mixing stopped-flow optical spectroscopy at its absorption maximum of 420 nm. These results provide for the first time direct monitoring of the hydroxylation of a methane-derived substrate in the MMOH reaction pathway and demonstrate that Q decay occurs concomitantly with substrate consumption. PMID- 16220909 TI - Synthesis, structure, bridge-terminal exchange kinetics, and molecular orbital calculations of pyrazolate-bridged digallium complexes containing bridging phenyl groups. AB - Gallium complexes containing bridging phenyl groups were prepared and characterized. Treatment of triphenylgallium with 3,5-dimethylpyrazole, 3,5 diphenylpyrazole, or 3,5-di-tert-butylpyrazole in a 2:1 stoichiometry afforded the phenyl-bridged complexes (C6H5)2Ga(mu-Me2pz)(mu-C6H5)Ga(C6H5)2 (62%), (C6H5)2Ga(mu-Ph2pz)(mu-C6H5)Ga(C6H5)2.C7H8 (62%), or (C6H5)2Ga(mu-tBu2pz)(mu C6H5)Ga(C6H5)2 (40%), respectively, as colorless or off-white crystalline solids. These complexes were characterized by spectral and analytical methods, X-ray crystallography, bridge-terminal exchange kinetics, and molecular orbital calculations for simplified models. The molecular structure of (C6H5)2Ga(mu Me2pz)(mu-C6H5)Ga(C6H5)2 consists of a dimethylpyrazolato ligand with a diphenylgallium group bonded to each nitrogen atom. A phenyl group acts as a bridge between the two gallium atoms. The kinetics of bridge-terminal phenyl exchange was determined by 13C NMR spectroscopy between -30 and +30 degrees C, and afforded the following range of activation parameters: DeltaH = 6.0-8.9 kcal/mol, DeltaS = -23.1 to -32.0 eu, and DeltaG(298) = 15.5-15.8 kcal/mol. The large, negative values of DeltaS imply ordered transition states relative to the ground state, and rotation along the N-GaPh3 vector without gallium-nitrogen bond cleavage. Molecular orbital calculations were conducted at the B3LYP/6-311G(d,p) level of theory on the simplified model H2Ga(mu-pz)(mu-C6H5)GaH2. The predicted out-of-plane phenyl group orientation arises from electronic interactions, in which hybridized orbitals on the phenyl group create delocalized molecular orbitals. However, the energy difference between a planar Ga2N2C ring and one with the bent carbon atom is only 1.77 kcal/mol, implying that the molecular orbitals provide little stabilization to the out-of-plane phenyl ligand. The combined results suggest that the close proximity of the gallium atoms is the principal determinant of the bridging phenyl interactions, and that complexes of the heavier group 13 elements with bridging hydrocarbon ligands are likely to be more accessible than the current state of the literature would suggest. PMID- 16220910 TI - A comparison of crystalline- and graft polymer-based chemosensors. AB - Herein, we report a highly sensitive luminescent thin film chemosensor constructed out of a small-molecule donor/acceptor system. Two types of films were compared: one using a small-molecule crystalline donor/acceptor pair and the other using a donor-graft polymer/small-molecule acceptor pair. The acceptor selected for this proof of concept responds to acid, causing its absorption and emission bands to red-shift, which increases spectral overlap with the donor. This increase in overlap greatly enhances energy transfer from the acceptor to the donor. Signal amplification was ascertained by measuring the ratio of acceptor fluorescence when the donor was excited versus direct excitation of the acceptor. Both types of films exhibited large amplification. For the polymeric system, the mechanism of energy migration was investigated by the use of steady state fluorescence spectroscopy. The mechanism was determined to be dominated by an exciton-hopping process. PMID- 16220911 TI - Synthesis and characterization of the first carbene derivative of a polyoxometalate. AB - Reaction of the Keggin-type polyanion [PW11O39]7- with the tetrakis-carbene ruthenium precursor [RuLMe4Cl2] (LMe = 1,3-dimethylimidazolidine-2-ylidene), in water, results in the formation of Na4K9[(PW9O34)2(cis-WO2)(cis-RuLMe2)].23H2O, which is the first carbene derivative of a polyoxometalate. The oxidation state of the ruthenium is confirmed by XANES experiments. PMID- 16220912 TI - Temperature dependence of NMR order parameters and protein dynamics. AB - The helical subdomain, HP36, of the F-actin-binding headpiece domain of chicken villin, is the smallest naturally occurring polypeptide that folds to a thermostable compact structure. Unconstrained molecular dynamics simulations and constrained molecular dynamics simulations using umbrella sampling are used to study the temperature dependence of internal motions of the backbone amide moieties of HP36. The potential of mean force (PMF) for the N-H bond vector, determined from the constrained simulations, is found to be temperature dependent. A simple analytical expression is derived that describes the temperature dependence of the PMF. The parameters of this model are obtained from the PMF, from the unconstrained molecular dynamics simulations, or from experimental values of the generalized order parameter. The results provide a linkage between experimental and theoretical measures of the temperature dependence of protein motions. PMID- 16220913 TI - Dynamics of curved DNA molecules: prediction and experiment. AB - We have developed a quantitative predictive model capable of describing the dynamics of migration of intrinsically curved DNA fragments on polyacrylamide gels. The model takes into account structural features of DNA, end-to-end distance, screening of hydrodynamic interactions, ionic strength of buffer, electrostatic persistence length, structural fluctuations of the macromolecule, counter condensation, and variation of dielectric constant and viscosity of water with MPD. In doing so, we have also addressed a decade old issue on the effect of the organic solvent 2-methyl-2,4-pentanediol on gel migration of phased A-tracts. We show here that A-tract-solvent interactions are less favored compared with A tract-A-tract and solvent-solvent interactions. PMID- 16220914 TI - Solution and solid-state spin-crossover behavior in a pseudotetrahedral d7 ion. AB - This Communication describes a pseudotetrahedral d7 complex, [PhBP3]Co(OSiPh3), that exhibits thermally induced spin-crossover both in solution and in the solid state. Magnetic crossover behavior is achieved by confluence of the X-type ligand and the tripodal auxiliary employed. Four-coordinate platforms of the present geometry type may offer a new approach to magnetic spin-crossover behavior distinct from their electronically related pseudooctahedral counterparts. PMID- 16220915 TI - Cell surface labeling of Escherichia coli via copper(I)-catalyzed [3+2] cycloaddition. AB - Labeling of the cell surface of Escherichia coli was accomplished by expression of a recombinant outer membrane protein, OmpC, in the presence of the unnatural amino acid azidohomoalanine, which acts as a methionine surrogate. The surface exposed azide moieties of whole cells were biotinylated via Cu(1)-catalyzed [3+2] azide-alkyne cycloaddition. The specificity of labeling of both wild-type OmpC and a mutant containing additional methionine sites for azidohomoalanine incorporation was confirmed by Western blotting. Flow cytometry was performed to examine the specificity of the labeling. Cells that express the mutant form of OmpC in the presence of azidohomoalanine, which were biotinylated and stained with fluorescent avidin, exhibit a mean fluorescence 10-fold higher than the background. Incorporation of an unnatural amino acid can thus be determined on a single-cell basis. PMID- 16220916 TI - Thermodynamics of phase transitions in langmuir monolayers observed by vibrational sum frequency spectroscopy. AB - Vibrational sum-frequency spectroscopy (VSFS) was used to study gauche defects in octadecylamine (ODA) monolayers at the air/water interface. The VSFS spectra provide unique insights into phase transitions that occur as a result of changes in the structure of the monolayer's hydrophobic region. These changes can be attributed to the increased presence of gauche conformers in the ODA alkyl chains during the monolayer's transition from the solid to liquid phase. Temperature dependent spectra from monolayers at several different pressures were used to assign the phase transition temperature based on the observed changes in microscopic structure. Through application of a two-dimensional form of the Clapeyron equation, the first in situ measurements of the entropy and enthalpy changes associated with gauche conformers in a monolayer were made. PMID- 16220917 TI - Identification of active-site inhibitors of MurG using a generalizable, high throughput glycosyltransferase screen. AB - MurG is a glycosyltransferase involved in the biosynthesis of bacterial peptidoglycan. It is a potentially important antibiotic target, but no inhibitors of the enzyme have been reported. In general, inhibitors of glycosyltransferases have been difficult to design. Furthermore, no glycosyltransferase inhibitors have been identified through high-throughput screening, perhaps because appropriate screens for glycosyltransferase inhibition have not been developed. In this manuscript, we describe the development of a high-throughput screen for MurG that was used to screen a 50 000 compound library for inhibitors. The screen, which can be generalized to other glycosyltransferases, led to the identification of a family of active-site directed MurG inhibitors. The family of inhibitors contains a five-membered heterocyclic core that appears to function as a diphosphate mimic with respect to the presentation of substituents. We discuss the implications of this result and the utility of the screen for identifying inhibitors of other glycosyltransferases. PMID- 16220918 TI - Screening of protein crystallization conditions on a microfluidic chip using nanoliter-size droplets. AB - Protein crystallization is a major bottleneck in determining tertiary protein structures from genomic sequence data. This paper describes a microfluidic system for screening hundreds of protein crystallization conditions using less than 4 nL of protein solution for each crystallization droplet. The droplets are formed by mixing protein, precipitant, and additive stock solutions in variable ratios in a flow of water-immiscible fluids inside microchannels. Each droplet represents a discrete trial testing different conditions. The system has been validated by crystallization of several water-soluble proteins. PMID- 16220919 TI - Full miscibility of disk- and rod-shaped mesogens in the nematic phase. AB - Mesogens containing rod-shaped moieties as well as one disk-shaped group exhibit nematic phase behavior and are entirely miscible with rod- and disk-shaped mesogens in the nematic phase. In the phase diagram, an extra nematic phase appears at nearly equimolar disk-rod ratios, where the nematic biaxial phase has been predicted. PMID- 16220920 TI - Enantioface-selective palladium-catalyzed silaboration of allenes via double asymmetric induction. AB - Enantioenriched beta-borylallylsilanes were synthesized by palladium-catalyzed enantioface-selective addition of the silicon-boron bond to terminal allenes using a palladium catalyst possessing a chiral monodentate phosphine ligand. Use of a silylborane bearing a chiral auxiliary on the boron atom was beneficial to gain enantioface selectivities as high as 96% de. PMID- 16220921 TI - Palladium-catalyzed alpha-arylation of esters and amides under more neutral conditions. AB - Two procedures for the alpha-arylation of carbonyl compounds under conditions that are more neutral than those of reactions of aryl halides with alkali metal enolates are reported. The first procedure rests upon the development of catalysts bearing the hindered pentaphenylferrocenyl di-tert-butylphosphine (Q phos) and the highly reactive dimeric Pd(I) complex {P(t-Bu)3]PdBr}2. By this procedure, zinc enolates prepared from alpha-bromo esters and amides react with aryl halides to form alpha-aryl esters and amides in high yields under mild conditions with 1-2 mol % catalyst and with remarkable functional group tolerance. By the second procedure, silyl ketene and silyl ketimine acetals react with aryl bromides in the presence of substoichiometric zinc fluoride, 1 mol % Pd(dba)2, and 2 mol % P(t-Bu)3 in DMF solvent at 80 degrees C. Reactions of zinc tert-butyl acetate and propionate enolates and trimethylsilyl ketene acetals of tert-butyl propionate and methyl isobutyrate with aryl bromides bearing electron donating and potentially reactive, base-sensitive electron-withdrawing groups and with pyridyl bromides are reported. In addition, the diastereoselective coupling of phenyl bromide with an imide enolate bearing the Evans auxiliary is reported, and this study shows that racemization of base-sensitive stereocenters does not occur during the coupling process under these more neutral conditions. PMID- 16220922 TI - Novel peptides bearing pyrene and coumarin units with or without beta cyclodextrin in their side chains exhibit intramolecular fluorescence resonance energy transfer. AB - Novel peptides bearing the pyrene/coumarin FRET pair in their side chains have been designed and synthesized. Peptide 1 having endogenous beta-cyclodextrin (beta-CD) in the side chain exhibits FRET in aqueous solution, indicating that coumarin, being accommodated into the CD cavity, is separated from pyrene. Guest induced quenching of the fluorophores in 1 indicates that coumarin, being excluded from the CD cavity, comes into close contact with pyrene. Peptide 2 shows FRET only after addition of external beta-CD that again reflects the idea that beta-CD surely caps the coumarin unit in its hydrophobic cavity, and, therefore, quenching of the fluorophores can be prevented in FRET peptide probes. With this strategy, various peptide-based FRET probes can be developed that would be useful for studying biological phenomena in living cells. PMID- 16220923 TI - Polar phase hydroformylation: the dramatic effect of water on mono- and dirhodium catalysts. AB - The addition of 30% water (by volume) to acetone creates a remarkably effective polar phase solvent system for a dicationic dirhodium tetraphosphine hydroformylation catalyst. The initial turnover frequency (TOF) increases by 265% (to 73 min-1) for the hydroformylation of 1-hexene relative to the initial TOF in pure acetone (20 min-1). The aldehyde linear to branched (L:B) ratio increases to 33:1, and alkene isomerization and hydrogenation side reactions are essentially eliminated. Comparisons with monometallic rhodium catalysts based on PPh3, Bisbi, Naphos, and Xantphos ligands demonstrate that this polar-phase bimetallic catalyst is one of the fastest and most selective hydroformylation systems known under these mild conditions (90 degrees C, 6.2 bar H2/CO). The monometallic catalysts also show rate enhancements (although considerably smaller) in water acetone, but Rh-Xantphos does show a large increase of 115%, with considerably reduced alkene isomerization side reactions. The dramatic effect of water on the dirhodium catalyst system is believed to be due to simple inhibition of the fragmentation of the catalytically active species into inactive mono- and bimetallic complexes. PMID- 16220924 TI - Unexpected Hofmann elimination in the benzophenone-(phenylthio)acetic tetrabutylammonium salt photoredox system. AB - The triplet state of benzophenone was quenched by the tetrabutylammonium salt of (phenylthio)acetic acid in acetonitrile solutions. The quenching event, following laser flash photolysis, resulted in the formation of a transient ion pair consisting of the benzophenone radical anion and the tetrabutylammonium cation. Subsequently this ion pair decays with the quaternary ammonium cation undergoing a Hofmann elimination to form butane-1 and tributylamine, which were detected in steady-state irradiation. This appears to be the first report of an ion pair consisting of a benzophenone radical anion and an organic cation (nonradical), in addition to the first report of a photoinduced Hofmann elimination in quaternary ammonium ions. PMID- 16220925 TI - Vibrational spectroscopy of CO in gas-phase rhodium cluster-CO complexes. AB - Infrared spectra of isolated unsaturated rhodium cluster-CO complexes in the region of the CO stretching vibration, nu(CO), are measured using a molecular beam depletion technique. These spectra provide benchmarks for interpreting values of nu(CO) that are found when CO is used to probe Rh surfaces and supported Rh nanoparticles. Supported nanoparticles have shifts of nu(CO) of as much as +100 cm-1 compared to the free clusters measured here, indicative of significant charge transfer to the support. PMID- 16220926 TI - Narrow (n,m)-distribution of single-walled carbon nanotubes grown using a solid supported catalyst. AB - Unusually structure-selective growth of single-walled carbon nanotubes (SWNTs) has been attained using a CVD method with a solid supported catalyst. In this method, CO feedstock disproportionates on silica-supported catalytic nanoclusters of Co that are formed in situ from mixed salts of Co and Mo. The nanotube products are analyzed by spectrofluorimetry to reveal distributions resolved at the level of individual (n,m) structures. Two structures, (6,5) and (7,5), together dominate the semiconducting nanotube distribution and comprise more than one-half of that population. The average diameter of produced SWNTs is only 0.81 nm, and a strong propensity is found favoring chiral angles near the armchair limit. PMID- 16220927 TI - Mechanism of chromyl chloride epoxidation. AB - Modern electronic structure methods including complete active space perturbation theory to second order (CASPT2), and a range of density functional (DFT) methods are used to characterize the intermediates and transition states associated with ethylene epoxidation by chromyl chloride. A novel zwitterionic direct addition pathway is characterized which, when coupled with a dinuclear complex, exothermically produces epoxide through a pathway with a small kinetic barrier. PMID- 16220928 TI - Extraction of monodisperse palladium nanoparticles from dendrimer templates. AB - We describe a strategy for extracting nearly size-monodisperse, 1.7-nm-diameter Pd nanoparticles from within dendrimer hosts. The approach involves two steps. First, an aqueous solution of dendrimer-encapsulated Pd nanoparticles is prepared, followed by the addition of a toluene solution containing an n alkanethiol. When the two-phase system is stirred, n-alkanethiols presumably self assemble onto the surface of the Pd nanoparticles, extract them from the dendrimer, and transport them to toluene phase. UV-vis spectroscopy indicates that the Pd nanoparticles, in the form of monolayer-protected clusters, are transported into the toluene phase. FT-IR spectroscopy shows that the dendrimers remain in the aqueous phase. High-resolution transmission electron microscopy shows that the average size and size distribution of the particles do not change during the extraction process. PMID- 16220930 TI - Interatomic distance measurement in solid-state NMR between a spin- 1/2 and a spin- 5/2 using a universal REAPDOR curve. AB - A universal curve for the solid-state NMR REAPDOR experiment on an isolated spin 1/2-spin-5/2 pair is proposed that provides a simple means to measure their interatomic distance. REAPDOR data were obtained at three separate REAPDOR experiments using different values of the rotor spinning frequency. All points were fitted simultaneously to the universal formula without a need for full density matrix calculations. The 13C-17O distance of 2.45 A was measured between the C6 carbon and the 17O label in a tyrosine sample. The error of 8% in the dipolar coupling (Dfit = 278 Hz) is well within the 15% theoretical tolerance of this curve. PMID- 16220929 TI - Reversible electrochemical detection of nonelectroactive polyions. AB - Selective extraction principles for the recognition of nonelectroactive polyions such as heparin and protamine exist, but the high ionic valency renders the extraction process irreversible. A response principle for the reversible detection of such polyions is proposed here. The extraction of the polyionic analyte to the membrane and its subsequent back-extraction is now controlled electrochemically. The principle is established with a protamine electrode, and excellent stability and reproducibility are demonstrated. This method has important implications for the design of chemical recognition principles for polyionic analytes. PMID- 16220931 TI - The first fluoride one-dimensional nanostructures: microemulsion-mediated hydrothermal synthesis of BaF2 whiskers. AB - A convenient microemulsion-mediated hydrothermal process was employed for the first synthesis of BaF2 whiskers with lengths up to 50 mum and aspects ratios as high as 1000, each of which is a single crystal with a growth direction of (111). A directed aggregation growth process mediated by the microemulsion droplet building blocks is proposed for the formation of BaF2 whiskers. PMID- 16220932 TI - Molecular logic: a half-subtractor based on tetraphenylporphyrin. AB - Spectroscopic changes from a solution of the amphophile 5,10,15,20 tetraphenylporphyrin in N,N-dimethylformamide using inputs of acid (HClaq) and base (KOBut) are interpreted as XOR and INHIBIT logic operations. The combination of these two operations gives rise to a device that acts as a molecular half subtractor. PMID- 16220933 TI - Rapid-freeze-quench magnetic circular dichroism of intermediate X in ribonucleotide reductase: new structural insight. AB - To elucidate the electronic structure of intermediate X in the oxygen activation reaction of the R2 subunit of ribonucleotide reductase, a protocol has been developed to perform magnetic circular dichroism (MCD) on a rapid-freeze-quench, strain free optical sample. RFQ-MCD data have been collected on intermediate X in the double mutant of R2, Y122/Y356F. While X has been reported to exhibit a broad absorption band at 365 nm, there are at least 10 electronic transitions observed at low-temperature MCD. From C0/D0 ratios, the transitions of X can be divided into three regions: 16 000-22 000 cm-1 region involving spin-allowed ligand field transitions of the Fe(IV), 23 000-24 000 cm-1 region of spin-forbidden, spin-flip transitions on the Fe(IV), and the charge transfer (CT) region from 26 000 to 32 000 cm-1. The C0/D0 ratios from d --> d and CT transitions strongly support significant Fe(IV) character coupled into the paramagnetic center. Ligand field (spin-allowed d --> d region) analysis allows the bis-mu-oxo and mu-oxo plus other monoanionic bridge possibilities for the structure of intermediate X to be distinguished, providing new insight into the molecular mechanism of the cluster formation in R2. PMID- 16220934 TI - Chemical contrasting in a single polymer molecule AFM experiment. AB - We developed a simple contrasting procedure to improve the AFM visualization of single positively charged polymer chains deposited on substrates of a relatively high roughness via the decoration of the molecules with hexacyanoferrate anions or negatively charged clusters of cyanide-bridged complexes. PMID- 16220935 TI - Enantioselective Michael additions to alpha,beta-unsaturated imides catalyzed by a Salen-Al complex. AB - (Salen)aluminum complex 1b is an efficient catalyst for the conjugate addition of di- and trisubstituted nitriles to a wide range of acyclic alkyl- and aryl substituted alpha,beta-unsaturated imides. This new methodology provides access to multifunctional compounds that previously have not been readily accessible in enantioenriched form. Synthetic applications of these products include the preparation of enantiomerically enriched piperidines, as exemplified by an expedient asymmetric catalytic synthesis of (-)-paroxetine. PMID- 16220936 TI - Enantioselective syntheses of aeruginosin 298-A and its analogues using a catalytic asymmetric phase-transfer reaction and epoxidation. AB - We developed a versatile synthetic process for aeruginosin 298-A as well as several attractive analogues, in which all stereocenters were controlled by a catalytic asymmetric phase-transfer reaction and epoxidation. Furthermore, drastic counteranion effects in phase-transfer catalysis were observed for the first time, making it possible to three-dimensionally fine-tune the catalyst (ketal part, aromatic part, and counteranion). PMID- 16220937 TI - Application of high pressure induced by water-freezing to the direct catalytic asymmetric three-component List-Barbas-Mannich reaction. AB - High pressure induced by water-freezing has been successfully applied to the direct catalytic asymmetric-three component List-Barbas-Mannich reaction, in which higher yield and better enantioselectivity can be realized than those from the reaction at room temperature under 0.1 MPa. PMID- 16220938 TI - A new catalytic conjugate addition/aldol strategy that avoids preformed metalated nucleophiles. AB - A new conjugate addition/aldol strategy has been developed. Unlike conventional procedures that require metalation of the nucleophilic species, the procedure allows the direct coupling of aryl iodides, aldehydes, and acrylates in a nickel catalyzed, organozinc-promoted process. PMID- 16220939 TI - The cyclooctatriene-eta2-ynyl potassium zwitterionic radical: evidence for a potassium organometallic. AB - Low-temperature (-120 degrees C) dehydrohalogenation of bromocyclooctatetraene (BrC8H7) with either sodium or potassium tert-butoxide followed by alkali metal reduction was used to generate the anion radical of [8]annulyne (C8H6*-) in tetrahydrofuran. EPR analysis at -120 degrees C reveals an extraordinarily large metal splitting when K or Cs (aK of 0.214 G and aCs of 3.26 G) serves as the reducing agent. The large aM is due to the metal cation interacting with the p orbitals, within the alkyne moiety, that are in the plane of the ring system. The ionic radius of K+ is 1.33 A, which is larger than the B3LYP predicted distance between carbons 1 and 2 (1.23 A). However, the ionic radius of Na+ is only 0.95 A, and it is too small to simultaneously interact with both p-orbitals. Hence, no aM is observed when Na (ordinarily aNa > aK) or Li serves as the reducing agent. After the addition of 18-crown-6 to either the K or the Cs reduced system, two anion radicals are present. One is the system where the 18-crown-6 encapsulated metal complex is normally ion paired over the face of the ring system and aM = 0. The other is the cyclooctatriene-eta2-ynyl 18-crown-6 encapsulated metal zwitterion radical exhibiting a large aM. The ion pair to organometallic equilibrium constant is 1.6 +/- 0.1 and 3.5 +/- 0.1 for the K and Cs systems, respectively. PMID- 16220940 TI - Spectroscopic and electronic structure studies of 2,3-dihydroxybiphenyl 1,2 dioxygenase: O2 reactivity of the non-heme ferrous site in extradiol dioxygenases. AB - The extradiol dioxygenase, 2,3-dihydroxybiphenyl 1,2-dioxygenase (DHBD, EC 1.13.11.39), has been studied using magnetic circular dichroism (MCD), variable temperature variable-field (VTVH) MCD, X-ray absorption (XAS) pre-edge, and extended X-ray absorption fine structure (EXAFS) spectroscopies, which are analogous to methods used in earlier studies on the extradiol dioxygenase catechol 2,3-dioxygenase [Mabrouk et al. J. Am. Chem Soc. 1991, 113, 4053-4061]. For DHBD, the spectroscopic data can be correlated to the results of crystallography and with the results from density functional calculations to obtain detailed geometric and electronic structure descriptions of the resting and substrate (DHB) bound forms of the enzyme. The geometry of the active site of the resting enzyme, square pyramidal with a strong Fe-glutamate bond in the equatorial plane, localizes the redox active orbital in an orientation appropriate for O(2) binding. However, the O(2) reaction is not favorable, as it would produce a ferric superoxide intermediate with a weak Fe-O bond. Substrate binding leads to a new square pyramidal structure with the strong Fe-glutamate bond in the axial direction as indicated by a decrease in the (5)E(g) and increase in the (5)T(2g) splitting. Electronic structure calculations provide insight into the relative lack of dioxygen reactivity for the resting enzyme and its activation upon substrate binding. PMID- 16220941 TI - Novel functionally grafted pseudo-semi-interpenetrating networks constructed by reactive linear-dendritic copolymers. AB - This paper describes the synthesis of amphiphilic pseudo-semi-interpenetrating polymer networks (pseudo-semi-IPNs) containing linear poly(styrene) and poly(ethylene glycol) (PEG) cross-linked through monodendritic fragments. A unique feature of the synthetic strategy is the permanent attachment of the linear segment to the PEG network by a transesterification reaction between the hydroxyl groups at both ends of the PEG and peripheral ethyl ester moieties in the monodendron portion of a linear poly(styrene)-dendritic poly(benzyl ether) AB block copolymer. The proceeding of the reaction is monitored by (1)H NMR and size exclusion chromatography (SEC). The formation of an interlock structure between the linear block and the network matrix in the pseudo-semi-IPN is evidenced by the results from spectroscopic analyses and differential scanning calorimetry measurements. The accessibility of functional centers in the grafted semi-IPN is confirmed by model reactions with fluorescent markers, fluorescence spectroscopy, and NMR techniques and shows the potential of these novel materials as sequestering reagents for resin capture-release applications in parallel synthesis, combinatorial chemistry, and advanced drug design. PMID- 16220942 TI - The first total synthesis of discorhabdin A. AB - The first stereoselective total synthesis of a potent antitumor alkaloid, discorhabdin A (1), which is a unique sulfur-containing pyrroloiminoquinone alkaloid, is described. The key step in the stereocontrolled total synthesis of 1 involves both a diastereoselective oxidative spirocyclization using a hypervalent iodine(III) reagent and an efficient construction of the labile and highly strained N,S-acetal skeleton. These methodologies provide a breakthrough in the total syntheses of these promising new antitumor agents, discorhabdins and their analogues, which should serve as valuable probes for structure-activity studies. PMID- 16220943 TI - Synthesis and photophysical studies of bis-enediynes as tunable fluorophores. AB - We have synthesized a family of bis-enediynes by two complementary Pd/Cu catalyzed Sonogashira cross-coupling methods. One is a modified Sonogashira reaction between the TMS-protected tetraalkyne 20 (or 21) and various aromatic bromides to afford bis-enediynes 22a-d and 23a-d bearing different peripheral aryl units. The other, the reaction of bifunctional 1,1-dibromo-1-alkenes with phenylacetylene, afforded a series of bis-enediynes 24-32 bearing various core aryl groups. These chemical modifications to the core and periphery of bis enediynes induce dramatic changes in absorption and emission spectra. Bis enediynes 22 and 23 show a large Stokes shift of about 50-110 nm when compared to the less-conjugated bis-enediynes 20 and 21. Absorptions and emissions of bis enediynes 25, 27-29, and 31 were red-shifted relative to those of enediyne 35. Substantial increases in fluorescence quantum yields are observed as a result of extending the pi-conjugation. The emission wavelength of bis-enediynes was tailored from indigo blue to reddish-orange, suggesting that the color of emission can be tunable by modification of the core and/or peripheral units. PMID- 16220944 TI - Thermally reversible formation of microspheres through non-covalent polymer cross linking. AB - Bis-thymine units were used to noncovalently cross-link a complementary diamidopyridine-functionalized copolymer. Upon combination in noncompetitive solvents, discrete micron-scale spherical aggregates were formed arising from specific three-point polymer-cross-linker hydrogen bonding interactions. The diameter of these microspheres could be controlled through spacer structure. The cross-linking process was fully thermally reversible, with complete dissolution observed at 50 degrees C and reformation of the aggregates upon return to ambient temperature. This process could be repeated multiply, with lower particle dispersity observed arising from the annealing process. PMID- 16220945 TI - Catalytic enantioselective conjugate reduction of lactones and lactams. AB - A dramatic acceleration of the enantioselective copper-catalyzed conjugate reduction of alpha,beta-unsaturated lactones, lactams, and esters is reported upon addition of alcohol additives. Good to excellent yields and enantioselectivities were realized using a catalyst generated in situ from CuCl(2).H(2)O, t-BuONa, p-tol-BINAP, and PMHS, and this methodology was applied to the synthesis of (-)-Paroxetine. PMID- 16220946 TI - Fabrication of nanoscale metallic spirals using phospholipid microtubule organizational templates. AB - We describe the fabrication of metallic Cu spiral/helical nanostructures prepared via selective electroless metallization of a phospholipid microtubule template. The metallization template is created through selective, sequential adsorption of the oppositely charged polyelectrolytes, sodium poly(styrenesulfonate) (PSS) and poly(ethyleneimine) (PEI), onto nanoscale seams naturally occurring on the microtubule surface. A negatively charged Pd(II) nanoparticle catalyst is bound to the terminal cationic PEI layer of the multilayer film and initiates selective template metallization to form the helical Cu nanostructures. Details of the process are presented, and a mechanism and factors affecting the control of the feature critical dimensions are discussed. PMID- 16220947 TI - Single-electron entrapment of [8]annulyne, biannulenylenes, and an annulenoannulene. AB - The low-temperature (-100 degrees C) dehydrohalogenation of bromocyclooctatetraene followed by immediate electron-transfer yields a stable solution of the [8]annulyne anion radical. If the unstable [8]annulyne is reacted with itself, cyclobutadiene, or benzyne, the respective bi-[8]annulenylene, [6]annuleno[8]annulene, or [6]-[8]annulenylene can be trapped as their anion radicals via one-electron transfer. These condensation products were all obtained from simple [2 + 2] cycloaddition reactions. B3LYP/6-31G geometry optimizations were carried out, and the calculated spin densities were compared to the EPR spectral results obtained for the anion radicals of [6]annuleno[8]annulene, [8]annulyne, bi[8]annulenylene, and [6]-[8]annulenylene, and excellent agreement has been realized. This simple "one-pot" approach should be applicable to a wide range of such systems. PMID- 16220948 TI - New insight into modeling non-covalently imprinted polymers. AB - Three series of polymers were carefully formulated with increasing amounts of template while keeping the polymer components constant. The number of binding sites (N) and the number average association constant (K(n)()) were calculated for each polymer in a series, using equations adapted from the literature describing molecularly imprinted polymers (MIPs). The trends of N and K(n)() for each series of polymers, which were graphed versus percent template, suggest multiple functional monomers in the binding sites of noncovalent MIPs. This new insight has implications for understanding the underlying mechanisms for the formation of binding sites in the MIPs studied. PMID- 16220949 TI - Development and application of a new general method for the asymmetric synthesis of syn- and anti-1,3-amino alcohols. AB - A general method is described for asymmetric synthesis of both syn- and anti-1,3 amino alcohols. The first application of metalloenamines derived from N-sulfinyl imines is reported for the highly diastereoselective addition to aldehydes. The reduction of the product beta-hydroxy N-sulfinyl imines 2 with catecholborane and LiBHEt(3) provides syn- and anti-1,3-amino alcohols with very high diastereomeric ratios. This method was found to be effective for a variety of substrates incorporating either aromatic or various aliphatic substituents. The convergent and efficient asymmetric syntheses of the two natural products, (-)-8 epihalosaline and (-)-halosaline, were also accomplished. PMID- 16220950 TI - Infrared-visible sum frequency generation spectroscopic study of molecular orientation at polystyrene/comb-polymer interfaces. AB - Surface-sensitive infrared-visible sum frequency generation spectroscopy (SFG) in total internal reflection geometry has been used to study the structure of poly(vinyl n-octadecyl carbamate-co-vinyl acetate) (PVNODC) or poly(octadecyl acrylate) (PA-18) in contact with a deuterated or hydrogenated polystyrene (dPS or hPS) layer. SFG spectra from the PVNODC (or PA-18)/hPS interface show methyl and methylene peaks corresponding to PVNODC (or PA-18) and phenyl peaks corresponding to the PS. Analysis suggests that the methyl groups are tilted at angles less than 30 degrees with respect to the surface normal. The presence of a strong methylene peak suggests the PVNODC alkyl side chains contain more gauche defects at the PS/PVNODC interface in comparison to PVNODC (or PA-18)/air interfaces. On heating, the SFG intensity from the PS/PA-18 interface drops sharply near the bulk melting temperature (T(m)) of PA-18. Interestingly, a similar drop in SFG signal is also observed for the PS phenyl groups. This demonstrates the ability of the phenyl group at the PS/PA-18 interface to rearrange itself upon the solid-to-liquid transition of the PA-18 alkyl side chain, at a temperature well below the bulk PS glass transition temperature. For PS/PVNODC interfaces, the drop in SFG intensity is gradual and in agreement with the three broad transitions of PVNODC observed in the bulk. PMID- 16220951 TI - Controlled and stereoselective polymerization of lactide: kinetics, selectivity, and microstructures. AB - Chiral aluminum isopropoxides based on enantiopure or racemic cyclohexylsalen ligand (Jacobsen ligand) have been prepared and employed for stereoelective/stereoselective ring-opening polymerization of lactide in toluene at 70 degrees C. The kinetics, selectivity of the catalysts, and microstructure of the obtained polylactides, using different combinations of lactide enantiomers and catalysts, were determined. In all cases, polylactides of controlled molecular weight, low polydispersity, and defined end groups were obtained. The polymerizations are first-order in both monomer(s) and catalyst. (R,R) CyclohexylsalenAlO(i)()Pr [(R,R)-1] polymerizes l-lactide significantly faster than d-lactide with a rate constant ratio k(l)/k(d) of approximately 14. The polymerization of rac-lactide using (R,R)-1 yields crystalline polymers, for which a selectivity factor of approximately 5.5 could be calculated up to 50% conversion based on the optical purity of the isolated polymers. The polymerization of a l-lactide/d-lactide (molar ratio: 80/20) mixture by (R,R)-1 furnishes an isotactic-atactic block copolylactide, which is highly crystalline with a T(m) = approximately 155 degrees C. Polymerization of rac-lactide applying rac-cyclohexylsalenAlO(i)()Pr [rac-1] yields isotactic stereoblock polylactides with a high T(m) = approximately 185 degrees C and a high degree of crystallinity. PMID- 16220952 TI - Mesoporous polyhedral cages and shells formed by textured self-assembly of ZnO nanocrystals. AB - We report a new structure, mesoporous structured polyhedral drum and spherical cages and shells formed by textured self-assembly of ZnO nanocrystals, which are made by a novel self-assembly process during epitaxial surface oxidation. The cages/shells exhibit unique geometrical shapes, and their walls are composed of mesoporous and textured ZnO nanocrystals. The structures of the cages and shells are studied, and a growth mechanism is proposed to be a process following solidification of the Zn liquid droplets, surface oxidation, and sublimation. PMID- 16220953 TI - Epitaxial heterostructures: side-to-side Si-ZnS, Si-ZnSe biaxial nanowires, and sandwichlike ZnS-Si-ZnS triaxial nanowires. AB - Epitaxial semiconducting heterostructures: side-to-side Si-ZnS, Si-ZnSe biaxial nanowires, and sandwichlike ZnS-Si-ZnS triaxial nanowires were grown via a simple two-stage thermal evaporation of mixed SiO and ZnS or SiO and ZnSe powders under a precise temperature control. Each nanowire had a uniform diameter of 40-120 nm and length ranging from several to several tens of micrometers. Subnanowires of Si, ZnS, and ZnSe within them had a diameter of 20-50, 40-60, and 20-50 nm, respectively. The optical property (nanoscale cathodoluminescence) was also investigated from these new structures. It is proposed that the Si nanowires formed through disproportionation of SiO to Si in the first evaporation stage and then served as one-dimensional nanoscale substrates (or templates) for an epitaxial growth of ZnS or ZnSe nanowires in the following thermal evaporation of ZnS or ZnSe powders. The present results suggest that the simple method might be useful for the synthesis of many other heterostructures containing Si and II-VI or III-V semiconducting composite nanowires to meet the growing demands of nanoscale science and technology. PMID- 16220955 TI - Single-wall carbon nanotube interaction with gases: sample contaminants and environmental monitoring. AB - Here, we show that residual contaminants in purified single-wall carbon nanotube bundles may be responsible for the reported sensitivity of the electronic and transport properties to oxygen. Removal of these contaminants makes the electronic spectra insensitive to O(2), CO, H(2)O, and N(2), while a strong sensitivity to NO(2), SO(2), and NH(3) is observed, confirming the possible application of single-wall nanotubes as powerful sensors capable of measuring environmentally significant levels of toxic gases. PMID- 16220954 TI - Spectroscopic investigation of stellacyanin mutants: axial ligand interactions at the blue copper site. AB - Detailed electronic and geometric structural descriptions of the blue copper sites in wild-type (WT) stellacyanin and its Q99M and Q99L axial mutants have been obtained using a combination of XAS, resonance Raman, MCD, EPR, and DFT calculations. The results show that the origin of the short Cu-S(Cys) bond in blue copper proteins is the weakened axial interaction, which leads to a shorter (based on EXAFS results) and more covalent (based on S K-edge XAS) Cu-S bond. XAS pre-edge energies show that the effective nuclear charge on the copper increases going from O(Gln) to S(Met) to no axial (Leu) ligand, indicating that the weakened axial ligand is not fully compensated for by the increased donation from the thiolate. This is further supported by EPR results. MCD data show that the decreased axial interaction leads to an increase in the equatorial ligand field, indicating that the site acquires a more trigonally distorted tetrahedral structure. These geometric and electronic structural changes, which result from weakening the bonding interaction of the axial ligand, allow the site to maintain efficient electron transfer (high H(DA) and low reorganization energy), while modulating the redox potential of the site to the biologically relevant range. These spectroscopic studies are complemented by DFT calculations to obtain insight into the factors that allow stellacyanin to maintain a trigonally distorted tetrahedral structure with a relatively strong axial Cu(II)-oxygen bond. PMID- 16220956 TI - Nanopatterning of alkynes on hydrogen-terminated silicon surfaces by scanning probe-induced cathodic electrografting. AB - The electrochemical cathodic electrografting reaction, previously demonstrated on bulk silicon surfaces, can be patterned on the nanoscale utilizing conducting probe atomic force microscopy (CP-AFM). Alkyne electrografting is a particularly useful chemical technique since it leads to direct covalent attachment of conjugated alkynes to silicon. In addition, application of a forward bias during the reaction renders the surface less sensitive to oxidation and the resulting monolayers are very stable in air and basic aqueous solution. Alkyne monolayer lines can be drawn down to 40 nm resolution using a Pt-coated AFM tip, and the heights of the monolayers scale with the molecular length of the alkyne. The tip is biased (+) and the surface is biased (-) to drive the cathodic electrografting reaction under ambient conditions. The resistance of the monolayers to fluoride, as well as friction force microscopy, indicate that the alkynes are covalently bonded to the surface, not oxide-based, and hydrophobic. The reaction does not work with alkenes, and therefore hydrosilylation is not the primary mode of reaction. Wider lines (300 nm) can be produced using broadened Pt-coated AFM tips. This reaction could be important for the interfacing of conjugated molecules directly to silicon in a spatially controlled fashion. PMID- 16220957 TI - How does huperzine A enter and leave the binding gorge of acetylcholinesterase? Steered molecular dynamics simulations. AB - The entering and leaving processes of Huperzine A (HupA) binding with the long active-site gorge of Torpedo californica acetylcholinesterase (TcAChE) have been investigated by using steered molecular dynamics simulations. The analysis of the force required along the pathway shows that it is easier for HupA to bind to the active site of AChE than to disassociate from it, which for the first time interprets at the atomic level the previous experimental result that unbinding process of HupA is much slower than its binding process to AChE. The direct hydrogen bonds, water bridges, and hydrophobic interactions were analyzed during two steered molecular dynamics (SMD) simulations. Break of the direct hydrogen bond needs a great pulling force. The steric hindrance of bottleneck might be the most important factor to produce the maximal rupture force for HupA to leave the binding site but it has a little effect on the binding process of HupA with AChE. Residue Asp72 forms a lot of water bridges with HupA leaving and entering the AChE binding gorge, acting as a clamp to take out HupA from or put HupA into the active site. The flip of the peptide bond between Gly117 and Gly118 has been detected during both the conventional MD and SMD simulations. The simulation results indicate that this flip phenomenon could be an intrinsic property of AChE and the Gly117-Gly118 peptide bond in both HupA bound and unbound AChE structures tends to adopt the native enzyme structure. At last, in a vacuum the rupture force is increased up to 1500 pN while in water solution the greatest rupture force is about 800 pN, which means water molecules in the binding gorge act as lubricant to facilitate HupA entering or leaving the binding gorge. PMID- 16220958 TI - A highly active zinc catalyst for the controlled polymerization of lactide. AB - We report the preparation, structural characterization, and detailed lactide polymerization behavior of a new Zn(II) alkoxide complex, (L(1)ZnOEt)(2) (L(1) = 2,4-di-tert-butyl-6-{[(2'-dimethylaminoethyl)methylamino]methyl}phenolate). While an X-ray crystal structure revealed the complex to be dimeric in the solid state, nuclear magnetic resonance and mass spectrometric analyses showed that the monomeric form L(1)ZnOEt predominates in solution. The polymerization of lactide using this complex proceeded with good molecular weight control and gave relatively narrow molecular weight distribution polylactide, even at catalyst loadings of <0.1% that yielded M(n) as high as 130 kg mol(-)(1). The effect of impurities on the molecular weight of the product polymers was accounted for using a simple model. Detailed kinetic studies of the polymerization reaction enabled integral and nonintegral orders in L(1)ZnOEt to be distinguished and the empirical rate law to be elucidated, -d[LA]/dt = k(p)[L(1)ZnOEt][LA]. These studies also showed that L(1)ZnOEt polymerizes lactide at a rate faster than any other Zn-containing system reported previously. This work provides important mechanistic information pertaining to the polymerization of lactide and other cyclic esters by discrete metal alkoxide complexes. PMID- 16220959 TI - A general model for selectivity in olefin cross metathesis. AB - In recent years, olefin cross metathesis (CM) has emerged as a powerful and convenient synthetic technique in organic chemistry; however, as a general synthetic method, CM has been limited by the lack of predictability in product selectivity and stereoselectivity. Investigations into olefin cross metathesis with several classes of olefins, including substituted and functionalized styrenes, secondary allylic alcohols, tertiary allylic alcohols, and olefins with alpha-quaternary centers, have led to a general model useful for the prediction of product selectivity and stereoselectivity in cross metathesis. As a general ranking of olefin reactivity in CM, olefins can be categorized by their relative abilities to undergo homodimerization via cross metathesis and the susceptibility of their homodimers toward secondary metathesis reactions. When an olefin of high reactivity is reacted with an olefin of lower reactivity (sterically bulky, electron-deficient, etc.), selective cross metathesis can be achieved using feedstock stoichiometries as low as 1:1. By employing a metathesis catalyst with the appropriate activity, selective cross metathesis reactions can be achieved with a wide variety of electron-rich, electron-deficient, and sterically bulky olefins. Application of this model has allowed for the prediction and development of selective cross metathesis reactions, culminating in unprecedented three component intermolecular cross metathesis reactions. PMID- 16220960 TI - Reactions of B atoms and clusters with NO: experimental and theoretical characterization of novel molecules containing B, N, and O. AB - Reactions of boron atoms and clusters with NO molecules in solid argon have been studied using matrix isolation infrared absorption spectroscopy. The reaction products were identified by isotopic substitution ((10)B, (11)B, (15)N(16)O, (14)N(18)O, and mixtures) and comparison with density functional calculations of isotopic frequencies. In solid argon, boron atoms spontaneously reacted with NO to form the insertion molecule NBO. The BNBO and OBNNO molecules were formed by the B and NO addition reactions to NBO. The linear BBNO and BBBNO nitrosyls also were formed spontaneously on annealing. These molecules photochemically rearranged to the more stable BNBO and BNBBO isomers, which have linear polyyne like structures. The photosensitive OBNNO molecule decomposed to form the NNBO(2) van der Waals complex. In addition, the novel OBON diradical was also formed on photolysis in high-concentration NO experiments. PMID- 16220961 TI - Temperature dependence of domain motions of calmodulin probed by NMR relaxation at multiple fields. AB - Interdomain motions of Ca(2+)-ligated calmodulin were characterized by analyzing the nuclear magnetic resonance (15)N longitudinal relaxation rate R(1), transverse relaxation rate R(2), and steady-state {(1)H}-(15)N NOE of the backbone amide group at three different magnetic field strengths (18.8, 14.1, and 8.5 T) and four different temperatures (21, 27, 35, and 43 degrees C). Between 35 and 43 degrees C, a larger than expected change in the amplitude and the time scale of the interdomain motion for both N- and C-domains was observed. We attribute this to the shift in population of four residues (74-77) in the central linker from predominantly helical to random coil in this temperature range. This is consistent with the conformation of these residues in the calmodulin-peptide complex, where they are nonhelical. The doubling of the disordered region of the central helix (residues 78-81 at room temperature) when temperature is raised from 35 to 43 degrees C results in larger amplitude interdomain motion. Our analysis of the NMR relaxation data quantifies subtle changes in the interdomain dynamics and provides an additional tool to monitor conformational changes in multidomain proteins. PMID- 16220962 TI - Single-scan NMR spectroscopy at arbitrary dimensions. AB - Multidimensional nuclear magnetic resonance (NMR) provides one of the foremost analytical tools available to elucidate the structure and dynamics of complex molecules in their native states. Executing this kind of experiment generally requires collecting an n-dimensional time-domain signal S, from which the spectrum arises via an appropriate Fourier analysis of its various time variables. This time-domain signal is actually measured directly only along one of the time axes, while the effects introduced by the remaining time variables are monitored via a parametric incrementation of their values throughout independent experiments. Two-dimensional (2D) NMR experiments thus usually require longer acquisition times than unidimensional experiments, 3D NMR is orders-of-magnitude more time consuming than 2D spectroscopy, etc. Very recently, we proposed and demonstrated an approach whereby data acquisition in 2D NMR can be parallelized, enabling the collection of complete 2D spectral sets within a single transient. The present paper discusses the extension of this 2D NMR methodology to an arbitrary number of dimensions. The principles of the ensuing ultrafast n-dimensional NMR approach are described, and a variety of homo- and heteronuclear 3D and 4D NMR spectra collected within a fraction of a second are presented. PMID- 16220963 TI - Reaction of phenyl radicals with acetylene: quantum chemical investigation of the mechanism and master equation analysis of the kinetics. AB - The mechanism of the C(6)H(5) + C(2)H(2) reaction has been investigated by various quantum chemical methods. Electrophilic addition to the CC triple bond is found to be the only important mode of phenyl radical attack on acetylene. The initially formed chemically activated C(6)H(5)C(2)H(2) adducts may follow several isomerization pathways in competition with collisional stabilization and H elimination. Thermochemistry of various decomposition and isomerization channels is evaluated by the G2M method. For key intermediates, the following standard enthalpies of formation have been deduced from isodesmic reactions: 94.2 +/- 2.0 kcal/mol (C(6)H(5)CHCH), 86.4 +/- 2.0 kcal/mol (C(6)H(5)CCH(2)), and 95.5 +/- 1.8 kcal/ mol (o-C(6)H(4)C(2)H(3)). The accuracy of theoretical predictions was examined through extensive comparisons with available experimental and theoretical data. The kinetics and product branching of the C(6)H(5) + C(2)H(2) reaction have been evaluated by weak collision master equation/Rice-Ramsperger Kassel-Marcus (RRKM) analysis of the truncated kinetic model including only kinetically important transformations of the isomeric C(8)H(7) radicals. Available experimental kinetic data can be quantitatively reproduced by calculation with a minor adjustment of the C(6)H(5) addition barrier from 3.7 to 4.1 kcal/mol. Our predicted total rate constant, k(R1) = (1.29 x 10(10))T(0.834) exp(-2320/T) cm(3) mol(-)(1) s(-)(1), is weakly dependent on P and corresponds to the phenylation process under combustion conditions (T > 1000 K). PMID- 16220964 TI - Mono- and dibridged isomers of Si2H3 and Si2H4: the true ground state global minima. Theory and experiment in concert. AB - Highly correlated ab initio coupled-cluster theories (e.g., CCSD(T), CCSDT) were applied on the ground electronic states of Si(2)H(3) and Si(2)H(4), with substantive basis sets. A total of 10 isomers, which include mono- and dibridged structures, were investigated. Scalar relativistic corrections and zero-point vibrational energy corrections were included to predict reliable energetics. For Si(2)H(3), we predict an unanticipated monobridged H(2)Si-H-Si-like structure (C(s), (2)A'') to be the lowest energy isomer, in constrast to previous studies which concluded that either H(3)Si-Si (C(s), (2)A'') or near-planar H(2)Si-SiH (C(1), (2)A) is the global minimum. Our results confirm that the disilene isomer, H(2)Si-SiH(2), is the lowest energy isomer for Si(2)H(4) and that it has a trans bent structure (C(2)(h), (1)A(g)). In addition to the much studied silylsilylene, H(3)Si-SiH, we also find that a new monobridged isomer H(2)Si-H-SiH (C(1), (1)A, designated 2c) is a minimum on the potential energy surface and that it has comparable stability; both isomers are predicted to lie about 7 kcal/mol above disilene. By means of Fourier transform microwave spectroscopy of a supersonic molecular beam, the rotational spectrum of this novel Si(2)H(4) isomer has recently been measured in the laboratory, as has that of the planar H(2)Si-SiH radical. Harmonic vibrational frequencies as well as infrared intensities of all 10 isomers were determined at the cc-pVTZ CCSD(T) level. PMID- 16220965 TI - Electrocatalytic reactions mediated by N,N,N',N'-Tetraalkyl-1,4-phenylenediamine redox liquid microdroplet-modified electrodes: chemical and photochemical reactions In, and At the surface of, femtoliter droplets. AB - The electro-oxidation of electrolytically unsupported ensembles of N,N-diethyl N',N'-dialkyl-para-phenylenediamine (DEDRPD, R = n-butyl, n-hexyl, and n-heptyl) redox liquid femtoliter volume droplets immobilized on a basal plane pyrolytic graphite electrode is reported in the presence of aqueous electrolytes. Electron transfer at these redox liquid modified electrodes is initiated at the microdroplet-electrode-electrolyte three-phase boundary. Dependent on both the lipophilicity of the redox oil and that of the aqueous electrolyte, ion uptake into or expulsion from the organic deposits is induced electrolytically. In the case of hydrophobic electrolytes, redox-active ionic liquids are synthesized, which are shown to catalyze the oxidation of l-ascorbic acid over the surface of the droplets. In contrast, the photoelectrochemical reduction of the anaesthetic reagent halothane proceeds within the droplet deposits and is mediated by the ionic liquid precursor (the DEDRPD oil). PMID- 16220966 TI - Catalytic reduction of acetone by [(bpy)Rh]+: a theoretical mechanistic investigation and insight into cooperativity effects in this system. AB - Lahav, Milstein, and co-workers reported that the complex [(bpy)Rh(hd)](+)PF(6)( ) (bpy = substituted bipyridine ligand, hd = 1,5-hexadiene) shows catalytic activity in the hydrogenation of acetone (Tollner, K. et al. Science 1997, 278, 2100). The activity in an ordered monolayer was found to be dramatically greater than in solution. We used the DFT functional mPW1K (Lynch, B. J. et al. J. Phys. Chem. A 2000, 104, 4811) to investigate the mechanism of the homogenous reaction. The suitability of the mPW1K functional was verified by coupled cluster calculations on a model system. Bulk solvent effects were considered. Various alternative catalytic cycles were evaluated, and we found that one potential mechanism involves metal-catalyzed keto-enol tautomerization to form [(bpy)Rh(enol)](+) that adds hydrogen yielding a complex with axial and equatorial hydride ligands. The reaction continues via transfer of the hydrides to the enolic C=C bond thereby forming 2-propanol and regenerating the catalyst. Another potential catalytic cycle involves formation of [(bpy)Rh(acetone)(2)(H)(2)](+), which has a spectator solvent ligand, and initial transfer of the equatorial hydride to the carbonyl carbon of acetone. Other mechanisms involving hydrogen transfer to the acetone tautomer involved higher barriers. With an eye toward modeling multi-center catalysis, various model systems for the bpy ligand were considered. It was found that diimine (HN=CH CH=NH) compares very well with bpy, whereas cis-1,2-diiminoethylene (H(2)C=N CH=CH-N=CH(2)) yields a reaction profile very close to that of bpy. Finally, the system with two rhodium centers, [(diimine)Rh](2)(2+), was investigated. The results strongly suggest that an enol-type catalytic cycle occurs and that cooperativity between the two metal centers is responsible for the acceleration of the reaction in the monolayer system. PMID- 16220967 TI - On the viability of small endohedral hydrocarbon cage complexes: X@C4H4, X@C8H8, X@C8H14, X@C10H16, X@C12H12, AND X@C16H16. AB - Small hydrocarbon complexes (X@cage) incorporating cage-centered endohedral atoms and ions (X = H(+), H, He, Ne, Ar, Li(0,+), Be(0,+,2+), Na(0,+), Mg(0,+,2+)) have been studied at the B3LYP/6-31G(d) hybrid HF/DFT level of theory. No tetrahedrane (C(4)H(4), T(d)()) endohedral complexes are minima, not even with the very small hydrogen atom or beryllium dication. Cubane (C(8)H(8), O(h)()) and bicyclo[2.2.2]octane (C(8)H(14), D(3)(h)()) minima are limited to encapsulating species smaller than Ne and Na(+). Despite its intermediate size, adamantane (C(10)H(16), T(d)()) can enclose a wide variety of endohedral atoms and ions including H, He, Ne, Li(0,+), Be(0,+,2+), Na(0,+), and Mg(2+). In contrast, the truncated tetrahedrane (C(12)H(12), T(d)()) encapsulates fewer species, while the D(4)(d)() symmetric C(16)H(16) hydrocarbon cage (see Table of Contents graphic) encapsulates all but the larger Be, Mg, and Mg(+) species. The host cages have more compact geometries when metal atoms, rather than cations, are inside. This is due to electron donation from the endohedral metals into C-C bonding and C-H antibonding cage molecular orbitals. The relative stabilities of endohedral minima are evaluated by comparing their energies (E(endo)) to the sum of their isolated components (E(inc) = E(endo) - E(cage) - E(x)) and to their exohedral isomer energies (E(isom) = E(endo) - E(exo)). Although exohedral binding is preferred to endohedral encapsulation without exception (i.e., E(isom) is always exothermic), Be(2+)@C(10)H(16) (T(d)(); -235.5 kcal/mol), Li(+)@C(12)H(12) (T(d)(); 50.2 kcal/mol), Be(2+)@C(12)H(12) (T(d)(); -181.2 kcal/mol), Mg(2+)@C(12)H(12) (T(d)(); -45.0 kcal/mol), Li(+)@C(16)H(16) (D(4)(d)(); 13.3 kcal/mol), Be(+)@C(16)H(16) (C(4)(v)(); 31.8 kcal/mol), Be(2+)@C(16)H(16) (D(4)(d)(); -239.2 kcal/mol), and Mg(2+)@C(16)H(16) (D(4)(d)(); -37.7 kcal/mol) are relatively stable as compared to experimentally known He@C(20)H(20) (I(h)()), which has an E(inc) = 37.9 kcal/mol and E(isom) = -35.4 kcal/mol. Overall, endohedral cage complexes with low parent cage strain energies, large cage internal cavity volumes, and a small, highly charged guest species are the most viable synthetic targets. PMID- 16220969 TI - Designed multiple ligands. An emerging drug discovery paradigm. PMID- 16220970 TI - Structure-based design and discovery of protein tyrosine phosphatase inhibitors incorporating novel isothiazolidinone heterocyclic phosphotyrosine mimetics. AB - Structure-based design led to the discovery of novel (S)-isothiazolidinone ((S) IZD) heterocyclic phosphotyrosine (pTyr) mimetics that when incorporated into dipeptides are exceptionally potent, competitive, and reversible inhibitors of protein tyrosine phosphatase 1B (PTP1B). The crystal structure of PTP1B in complex with our most potent inhibitor 12 revealed that the (S)-IZD heterocycle interacts extensively with the phosphate binding loop precisely as designed in silico. Our data provide strong evidence that the (S)-IZD is the most potent pTyr mimetic reported to date. PMID- 16220971 TI - Selective inhibition of MCF-7(piGST) breast tumors using glutathione transferase derived 2-methylene-cycloalkenones. AB - Human glutathione (GSH) transferase (hGSTP1-1) catalyzes the conversion of antitumor 2-crotonyloxymethyl-2-cycloalkenones (COMCs) to highly reactive exocyclic enone alkylating agents. In vitro efficacy studies show that the cytotoxicities of the COMCs directly correlate with the level of expression of GSTP1-1 in MCF-7(piGST) versus MCF-7wt breast tumors, indicating that the exocyclic enones are the actual cytotoxic species. The COMCs are a potentially important new class of prodrugs, which can specifically target multi-drug resistant tumors overexpressing hGSTP1-1. PMID- 16220972 TI - Introduction of a cis-prolyl mimic in position 7 of the peptide hormone oxytocin does not result in antagonistic activity. AB - New insights into the structure-activity relationship of the peptide hormone oxytocin are presented. Incorporation of the novel cis-prolyl mimic 2,2-dimethyl 1,3-thiazolidine-4-carboxylic acid (pseudoproline, PsiPro) at position 7 of the hormone yielded the analogue [Cys(Psi(Me,Me)pro)]7oxytocin (1) that showed a 92 95% induction of the cis peptide bond conformation between Cys6 and PsiPro7, as determined by one- and two-dimensional NMR spectra in water and in DMSO-d6. The impact of the dimethyl moiety regarding conformation and bioactivity was investigated by the synthesis of the corresponding dihydro compound, [Cys(Psi(H,H)pro)]7oxytocin (2). Biological tests of the uterotonic activity, the pressor activity, and the binding affinity to the rat and human oxytocin receptors were carried out. As a most significant result, no antagonistic activities were found for both the cis-constrained analogue 1 and analogue 2, suggesting that a cis conformation between residues 6 and 7 of the molecule does not result in antagonistic activity. However, the about 10-fold reduction in agonistic activity of 1 as compared to oxytocin is consistent with the reduction of the trans conformation from 90% for oxytocin to 5-8% for compound 1. Compound 1 retained a high binding affinity for the oxytocin receptor, with K(i) values of 8.0 and 1.9 nM for the rat and the human receptor, respectively. The correlation between the biological activities and the cis contents obtained from NMR analysis for compounds 1, 2, and oxytocin leads to the hypothesis that a cis/trans conformational change plays an important role in oxytocin receptor binding and activation. PMID- 16220973 TI - G-protein-coupled receptor affinity prediction based on the use of a profiling dataset: QSAR design, synthesis, and experimental validation. AB - A QSAR model accounting for "average" G-protein-coupled receptor (GPCR) binding was built from a large set of experimental standardized binding data (1939 compounds systematically tested over 40 different GPCRs) and applied to the design of a library of "GPCR-predicted" compounds. Three hundred and sixty of these compounds were randomly selected and tested in 21 GPCR binding assays. Positives were defined by their ability to inhibit by more than 70% the binding of reference compounds at 10 microM. A 5.5-fold enrichment in positives was observed when comparing the "GPCR-predicted" compounds with 600 randomly selected compounds predicted as "non-GPCR" from a general collection. The model was efficient in predicting strongest binders, since enrichment was greater for higher cutoffs. Significant enrichment was also observed for peptidic GPCRs and receptors not included to develop the QSAR model, suggesting the usefulness of the model to design ligands binding with newly identified GPCRs, including orphan ones. PMID- 16220974 TI - Multiple-ligand-based virtual screening: methods and applications of the MTree approach. AB - We present a novel approach for ligand-based virtual screening by combining query molecules into a multiple feature tree model called MTree. All molecules are described by the established feature tree descriptor, which is derived from a topological molecular graph. A new pairwise alignment algorithm leads to a consistent topological molecular alignment based on chemically reasonable matching of corresponding functional groups. These multiple feature tree models find application in ligand-based virtual screening to identify new lead structures for chemical optimization. Retrospective virtual screening with MTree models generated for angiotensin-converting enzyme and the alpha1a receptor on a large candidate database yielded enrichment factors up to 71 for the first 1% of the screened database. MTree models outperformed database searches using single feature trees in terms of hit rates and quality and additionally identified alternative molecular scaffolds not included in any of the query molecules. Furthermore, relevant molecular features, which are known to be important for affinity to the target, are identified by this new methodology. PMID- 16220975 TI - Receptor flexibility in de novo ligand design and docking. AB - One of the major problems in computational drug design is incorporation of the intrinsic flexibility of protein binding sites. This is particularly crucial in ligand binding events, when induced fit can lead to protein structure rearrangements. As a consequence of the huge conformational space available to protein structures, receptor flexibility is rarely considered in ligand design procedures. In this work, we present an algorithm for integrating protein binding site flexibility into de novo ligand design and docking processes. The approach allows dynamic rearrangement of amino acid side chains during the docking and design simulations. The impact of protein conformational flexibility is investigated in the docking of highly active inhibitors in the binding sites of acetylcholinesterase and human collagenase (matrix metalloproteinase-1) and in the design of ligands in the S1' pocket of MMP-1. The results of corresponding simulations for both rigid and flexible binding sites are compared in order to gauge the influence of receptor flexibility in drug discovery protocols. PMID- 16220976 TI - Synthesis and antimuscarinic properties of quinuclidin-3-yl 1,2,3,4 tetrahydroisoquinoline-2-carboxylate derivatives as novel muscarinic receptor antagonists. AB - In the course of continuing efforts to develop potent and bladder-selective muscarinic M3 receptor antagonists, quinuclidin-3-yl 1-aryl-1,2,3,4 tetrahydroisoquinoline-2-carboxylate derivatives and related compounds were designed as conformationally restricted analogues of quinuclidin-3-yl benzhydrylcarbamate (8). Binding assays with rat muscarinic receptor subtypes revealed that the quinuclidin-3-yl 1-aryl-1,2,3,4-tetrahydroisoquinoline-2 carboxylate derivatives showed high affinities for the M3 receptor, and selectivity for the M3 receptor over the M2 receptor. Of these derivatives, (+) (1S,3'R)-quinuclidin-3'-yl 1-phenyl-1,2,3,4-tetrahydroisoquinoline-2-carboxylate monohydrochloride (9b) exhibited almost the same inhibitory activity against bladder contraction to that of oxybutynin (1), and more than 10-fold selectivity for bladder contraction versus salivary secretion, demonstrating that 9b may be useful for the treatment of symptoms associated with overactive bladder without having side effects such as dry mouth. PMID- 16220977 TI - Hydroxyethylene sulfones as a new scaffold to address aspartic proteases: design, synthesis, and structural characterization. AB - Hydroxyethylene sulfones were developed as novel scaffolds against aspartyl proteases. A diastereoselective synthesis has been established to introduce the required side chain decoration with desired stereochemistry. Depending on the substitution of the hydroxyethylene sulfone core, micro- to submicromolar inhibition of HIV-1 protease is achieved for the S-configuration at P1 and R configuration at the hydroxy-group-bearing backbone atom. This stereochemical preference is consistent with the S,R configuration of amprenavir. The racemic mixture of the most potent derivative (K(i) = 80 nM) was separated by chiral HPLC, revealing the S,R,S-enantiomer to be more active (K(i) = 45 nM). Docking studies suggested this isomer as the more active one. The subsequently determined crystal structure with HIV-1 protease, cocrystallized from a racemic mixture, exclusively reveals the S,R,S-enantiomer accommodated to the binding pocket. The transition state mimicking hydroxy group of the inhibitor is centered between both catalytic aspartates, while either its carbonyl or sulfonyl group forms H bonds to the structurally conserved water mediating interactions between ligand and Ile50NH/Ile50NH' of both flaps. Biological testing of the stereoisomeric hydroxyethylene sulfones against cathepsin D and beta-secretase did not reveal significant inhibition. Most likely, the latter proteases require inverted configuration at the hydroxy group. PMID- 16220978 TI - Angiotensin II pseudopeptides containing 1,3,5-trisubstituted benzene scaffolds with high AT2 receptor affinity. AB - Two 1,3,5-trisubstituted aromatic scaffolds intended to serve as gamma-turn mimetics have been synthesized and incorporated in five pseudopeptide analogues of angiotensin II (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe), replacing Val-Tyr-Ile, Val Tyr, or Tyr-Ile. All the tested compounds exhibited nanomolar affinity for the AT2 receptor with the best compound (3) having a K(i) of 1.85 nM. Four pseudopeptides were AT2 selective, while one (5) also exhibited good affinity for the AT1 receptor (K(i) = 30.3 nM). This pseudopeptide exerted full agonistic activity in an AT2 receptor induced neurite outgrowth assay but displayed no agonistic effect in an AT1 receptor functional assay. Molecular modeling, using the program DISCOtech, showed that the high-affinity ligands could interact similarly with the AT2 receptor as other ligands with high affinity for this receptor. A tentative agonist model is proposed for AT2 receptor activation by angiotensin II analogues. We conclude that the 1,3,5-trisubstituted benzene rings can be conveniently prepared and are suitable as gamma-turn mimics. PMID- 16220979 TI - Heteroaryl-substituted naphthalenes and structurally modified derivatives: selective inhibitors of CYP11B2 for the treatment of congestive heart failure and myocardial fibrosis. AB - Recently we proposed inhibition of aldosterone synthase (CYP11B2) as a novel strategy for the treatment of congestive heart failure and myocardial fibrosis. In this study the synthesis and biological evaluation of heteroaryl-substituted naphthalenes and quinolines (1-31) is described. Key step for the preparation of the compounds was a Suzuki cross-coupling. Activity of the compounds was determined in vitro using human CYP11B2 and selectivity was evaluated toward the human steroidogenic enzymes CYP11B1, CYP19, and CYP17. A large number of highly active and selective inhibitors of CYP11B2 was identified. The most active inhibitor was the 6-cyano compound 8 (IC50 = 3 nM) showing a competitive type of inhibition (K(i) value = 1.9 nM). The 6-ethoxy derivative 5 was found to be the most selective CYP11B2 inhibitor (IC50 = 12 nM; K(i) value = 8 nM; CYP11B1 IC50 = 5419 nM; selectivity factor = 451), showing no inhibition of human CYP3A4 (50 nM) and CYP2D6 (20 nM). Docking and molecular dynamics studies using our homology modeled CYP11B2 structure with selected compounds were performed. Caco-2 cell experiments revealed a large number of medium and highly permeable compounds and metabolic studies with 2 using rat liver microsomes showed sufficient stability. PMID- 16220980 TI - Discovery of iodinated somatostatin analogues selective for hsst2 and hsst5 with excellent inhibition of growth hormone and prolactin release from rat pituitary cells. AB - Inhibition of growth hormone (GH) and prolactin (PRL) release from the anterior pituitary gland is mediated through somatostatin receptor subtypes sst2 and sst5. It has been found that somatostatin (SS) analogues that are selective for both receptor subtypes are more effective at inhibiting GH and PRL release than monospecific analogues alone. We synthesized several disulfide-bridged octapeptide SS analogues. Iodinated compounds 7, (4-amino-3-iodo)-d-Phe-c[Cys-Tyr d-Trp-Lys-Val-Cys]-Thr-NH2, and 9, (4-amino-3-iodo)-d-Phe-c[Cys-(3-iodo)-Tyr-d Trp-Lys-Val-Cys]-Thr-NH2, were as potent as somatostatin in binding at receptors hsst2 and hsst5 and inhibited GH and PRL release from rat pituitary cells as potently as somatostatin. PMID- 16220981 TI - Improving the antiviral efficacy and selectivity of HIV-1 reverse transcriptase inhibitor TSAO-T by the introduction of functional groups at the N-3 position. AB - Novel derivatives of the anti-HIV-1 agent, TSAO-T, bearing at the N-3 position a variety of polar, lipophilic, or aromatic groups linked to that position through flexible polymethylene linkers of different length, were prepared and evaluated for their anti-HIV activity. Several compounds (within the series of polar bearing groups) exhibited a 2-6-fold improved antiviral potency with regard to the lead compound, TSAO-T. Moreover, some of the most active N-3 TSAO derivatives retain antiviral activity against the TSAO-T-resistant HIV-1 strain (Glu138 --> Lys). Interestingly, the N-methylcarboxamide derivative 17 was 5- to 6-fold more active (IC50: 0.56 microM) against recombinant HIV-1 reverse transcriptase than the lead compound, thus becoming the most active TSAO derivative synthesized so far. On the other hand, the N-3 methylcarboxamide TSAO derivative 12 turned out to have the highest selectivity index yet reported for this class of compounds (around > or =12 000). PMID- 16220982 TI - Investigation of the binding determinants of phosphopeptides targeted to the SRC homology 2 domain of the signal transducer and activator of transcription 3. Development of a high-affinity peptide inhibitor. AB - Signal transducer and activator of transcription 3 (Stat3) is a cytosolic transcription factor that relates signals from the cell membrane directly to the nucleus where it, in complex with other proteins, initiates the transcription of antiapoptotic and cell cycling genes, e.g., Bcl-x(L) and cyclin D1. In normal cells Stat3 transduces signals from cytokines such as IL-6 and growth factors such as the epidermal growth factor. Stat3 is constitutively activated in a number of human tumors. Antisense and dominant negative gene delivery result in apoptosis and reduced cell growth, thus this protein is an attractive target for anticancer drug design. As part of our research on the design of Src homology 2 (SH2) directed peptidomimetic inhibitors of Stat3, in this paper we describe structure-activity relationship studies that provide information on the nature of peptide-protein interactions of a high-affinity phosphopeptide inhibitor of Stat3 dimerization and DNA binding, Ac-Tyr(PO3H2)-Leu-Pro-Gln-Thr-Val-NH2, peptide 1. There is a hydrophobic surface on the SH2 domain that can accommodate lipophilic groups on the N-terminus. Of the amino acids tested, leucine provided the highest affinity at pY+1 and its main chain NH is involved with a hydrogen bond with Stat3, presumably Ser636. cis-3,4-Methanoproline is optimal as a backbone constraint at pY+2. The side chain amide protons of Gln are required for high affinity interactions. The C-terminal dipeptide, Thr-Val, can be replaced with groups ranging in size from methyl to benzyl. We synthesized a phosphopeptide incorporating groups that provided increases in affinity at each position. Thus, hydrocinnamoyl-Tyr(PO3H2)-Leu-cis-3,4-methanoPro-Gln-NHBn, 50, was the highest affinity peptide, exhibiting an IC50 of 125 nM versus 290 nM for peptide 1 in a fluorescence polarization assay. PMID- 16220983 TI - Ring systems in mutagenicity databases. AB - The distribution of ring systems in public mutagenicity databases is analyzed. An automated enumeration of substructures permits determination of the occurrence of different scaffolds in data sets. The counts are used to perform population analysis via proportions and odds ratios of mutagenic compounds. Pairwise calculations of odds ratios between scaffolds allow comparison of ring systems for isostere replacement studies. These findings are presented in tables that readily show which scaffold is likely to occur in mutagenic compounds. Also, rings identified in public domain mutagenicity data sets are compared to rings in drugs data sets; unfortunately, public mutagenicity data sets do not reflect the types of scaffolds in drugs and those typically used in medicinal chemistry. The findings bring into question the utility of predictive models that were derived from public domain data sets. The automated ring identification and statistical approaches used here can be applied to other pharmacological properties to yield information about chemical scaffolds. PMID- 16220984 TI - Antibiotic binding to monozinc CphA beta-lactamase from Aeromonas hydropila: quantum mechanical/molecular mechanical and density functional theory studies. AB - The active-site dynamics of apo CphA beta-lactamase from Aeromonas hydropila and its complex with a beta-lactam antibiotic molecule (biapenem) are simulated using a quantum mechanical/molecular mechanical (QM/MM) method and density functional theory (DFT). The quantum region in the QM/MM simulations, which includes the Zn(II) ion and its ligands, the antibiotic molecule, the catalytic water, and an active-site histidine residue, was treated using the self-consistent charge density functional tight binding (SCC-DFTB) model. Biapenem is docked at the active site unambiguously, based on a recent X-ray structure of an enzyme intermediate complex. The substrate is found to form the fourth ligand of the zinc ion with its 3-carboxylate oxygen and to hydrogen bond with several active site residues. The stability of the metal-ligand bonds and the hydrogen-bond network is confirmed by 500 ps molecular dynamics simulations of both the apo enzyme and the substrate-enzyme complex. The structure and dynamics of the substrate-enzyme complex provide valuable insights into the mode of catalysis in such enzymes that is central to the bacterial resistance to beta-lactam antibiotics. PMID- 16220985 TI - Development of nonsymmetrical 1,4-disubstituted anthraquinones that are potently active against cisplatin-resistant ovarian cancer cells. AB - A novel series of 1,4-disubstituted aminoanthraquinones were prepared by ipso displacement of 1,4-difluoro-5,8-dihydroxyanthraquinones by hydroxylated piperidinyl- or pyrrolidinylalkylamino side chains. One aminoanthraquinone (13) was further derivatized to a chloropropylamino analogue by treatment with triphenylphosphine-carbon tetrachloride. The compounds were evaluated in the A2780 ovarian cancer cell line and its cisplatin-resistant variants (A2780/cp70 and A2780/MCP1). The novel anthraquinones were shown to possess up to 5-fold increased potency against the cisplatin-resistant cells compared to the wild-type cells. Growth curve analysis of the hydroxyethylaminoanthraquinone 8 in the osteosarcoma cell line U-2 OS showed that the cell cycle is not frozen, rather there is a late cell cycle arrest consistent with the action of a DNA-damaging topoisomerase II inhibitor. Accumulative apoptotic events, using time lapse photography, indicate that 8 is capable of fully engaging cell cycle arrest pathways in G2 in the absence of early apoptotic commitment. 8 and its chloropropyl analogue 13 retained significant activity against human A2780/cp70 xenografted tumors in mice. PMID- 16220986 TI - Perhydroquinolylbenzamides as novel inhibitors of 11beta-hydroxysteroid dehydrogenase type 1. AB - High-throughput screening identified 5 as a weak inhibitor of 11beta-HSD1. Optimization of the structure led to a series of perhydroquinolylbenzamides, some with low nanomolar inhibitory potency. A tertiary benzamide is required for biological activity and substitution of the terminal benzamide with either electron-donating or -withdrawing groups is tolerated. The majority of the compounds show selectivity of >20 to >700-fold over 11beta-HSD2. Analogues which showed >50% inhibition of 11beta-HSD1 at 1 muM in an cellular assay were screened in an ADX mouse model. A maximal response of >70% reduction of liver corticosterone levels was observed for three compounds; 9m, 25 and 49. PMID- 16220987 TI - Synthesis and structure-activity relationships of beta- and alpha-piperidine sulfone hydroxamic acid matrix metalloproteinase inhibitors with oral antitumor efficacy. AB - alpha-Piperidine-beta-sulfone hydroxamate derivatives were explored that are potent for matrix metalloproteinases (MMP)-2, -9, and -13 and are sparing of MMP 1. The investigation of the beta-sulfones subsequently led to the discovery of hitherto unknown alpha-sulfone hydroxamates that are superior to the corresponding beta-sulfones in potency for target MMPs, selectivity vs MMP-1, and exposure when dosed orally. alpha-Piperidine-alpha-sulfone hydroxamate 35f (SC 276) was advanced through antitumor and antiangiogenesis assays and was selected for development. Compound 35f demonstrates excellent antitumor activity vs MX-1 breast tumor in mice when dosed orally as monotherapy or in combination with paclitaxel. PMID- 16220988 TI - Preliminary studies of new proteasome inhibitors in the tumor targeting approach: synthesis and in vitro cytotoxicity. AB - The proteasome is a multicatalytic protease that plays a critical role in the cell. The control of proteasomes could, thus, provide a weapon for the treatment of cancer. Therefore, we have synthesized six new peptide aldehyde inhibitors of the proteasome linked to the N-(2-diethylaminoethyl)benzamide (BZA-CO) structure, in order to target the cytotoxic activity to malignant melanoma cells. Biological studies demonstrated the influence of length and composition of the amino acid chain on the cytotoxicity of our compounds. Among them, compound 19 presents the highest cytotoxicity (IC50 = 0.64 +/- 0.07 micromol): this cytotoxicity was maintained in the presence of BZA-CO but decreased 8-fold compared to the control MG132. Fluorescence activated cell sorter (FACS) and cytotoxic activity analysis demonstrated the selectivity of compound 19 for melanoma cells. Finally, western blottings of ubiquitinated proteins in IPC227F cells as well as proteasome assays confirmed that the cytotoxicity was linked to an inhibition of the proteasome activity. PMID- 16220989 TI - Structure-activity relationship study on a simple cationic peptide motif for cellular delivery of antisense peptide nucleic acid. AB - Improving cellular uptake and biodistribution remains one of the major obstacles for a successful and broad application of peptide nucleic acids (PNAs) as antisense therapeutics. Recently, we reported the identification and functional characterization of an antisense PNA, which redirects splicing of murine CD40 pre mRNA. In this context, it was discovered that a simple octa(l-lysine) peptide covalently linked to the PNA is capable of promoting free uptake of the conjugate into BCL1 cells as well as primary murine macrophages. On the basis of this peptide motif, the present study aimed at identifying the structural features, which define effective peptide carriers for cellular delivery of PNA. While the structure-activity relationship study revealed some clear correlations, only a few modifications actually led to an overall improvement as compared to the parent octa(l-lysine) conjugate. In a preliminary PK/tissue distribution study in healthy mice, the parent conjugate exhibited relatively broad tissue distribution and only modest elimination via excretion within the time frame of the study. PMID- 16220990 TI - Glucosamine-glycerophospholipids that activate cell-matrix adhesion and migration. AB - Two new analogues derived from the platelet activating factor (PAF), containing glucosamine instead of the acetyl group, were synthesized, and their effect on the human keratinocyte cell line HaCaT was evaluated with respect to cytotoxicity, proliferation, adhesion, and migration. Starting with (R)-1,2 isopropylideneglycerol (3), the glycosylation acceptor 1-O-octadecyl-3-O-tert butyldimethylsilyl-sn-glycerol (6) was synthesized in three steps. Glycosylation of 6 with the already known O-(3,4,6-tri-O-acetyl-2-deoxy-2-dimethylmaleimido beta-D-glycopyranosyl)trichloracetimidate gave 1-O-octadecyl-2-O-(3',4',6'-tri-O acetyl-2'-deoxy-2'-dimethylmaleimido-beta-D-glucopyranosyl)-3-O-tert butyldimethylsilyl-sn-glycerol (7). After removing the (tert-butyldimethyl)silyl (TBDMS) group with FeCl3x6H2O, phosphoryl choline was introduced, yielding [1-O octadecyl-2-O-(2'-deoxy-2'-dimethylmaleimido-beta-D-glucopyranosyl)-sn glycero(3)]phosphorylcholine (2) (glucosimide-PAF). pH controlled cleavage of the amino protection group gave [1-O-octadecyl-2-O-(2'-deoxy-2'-amino-beta-D glucopyranosyl)-sn-glycero(3)]phosphorylcholine hydrochloride (1) (glucosamine PAF). 2 inhibited proliferation of HaCaT cells by 26% at nontoxic concentrations, while 1 increased the proliferation rate by 30% at low concentrations. At higher concentrations, both compounds showed cytotoxic properties with LD50 = 30 micromol/L (1) and LD50 = 5-6 micromol/L (2). Both 1 and 2 were potent promoters of cell adhesion and migration of HaCaT cells. PMID- 16220991 TI - A novel selective GABA(A) alpha1 receptor agonist displaying sedative and anxiolytic-like properties in rodents. AB - In our pursuit to identify selective ligands for Bz/GABA(A) receptor subtypes, a novel pyrazolo[1,5-a]pyrimidine derivative (4), the azaisostere of zolpidem, was synthesized and evaluated in vitro on bovine brain homogenate and on recombinant benzodiazepine receptors (alphaxbeta2/3gamma2, x = 1-3, 5) expressed in HEK293 cells. Compound 4 displayed affinity only for alpha1beta2gamma2 subtype (K(i) = 31 nM), and in an in-depth, in vivo study it revealed sedative and anxiolytic like properties without any amnesic and myorelaxant effects in rodents. PMID- 16220992 TI - Mechanisms of cytotoxicity of selected organogold(III) compounds. AB - The effects of a few cytotoxic organogold(III) compounds on ovarian A2780 human cancer cells were investigated in comparison to cisplatin and oxaliplatin. The tested compounds produced significant antiproliferative effects and promoted apoptosis to a greater extent than platinum drugs while causing only modest cell cycle modifications. The mechanistic implications of these findings are discussed: mitochondrial pathways are proposed to be directly involved in the apoptotic process in relation to selective inhibition of thioredoxin reductase. PMID- 16220994 TI - HIV vaccines in infants and children. AB - The interruption of HIV transmission from mother to child is important. Prevention strategies, including antiretroviral agents administered to the mother and/or child, can successfully prevent such transmission. Avoidance of breastfeeding or the administration of antiretroviral agents to the mother while she continues breastfeeding is another strategy. Based on the successful model of hepatitis B prevention by treatment of the newborn, research protocols have been designed to prevent mother-to-child transmission of HIV by the administration of passive HIV-specific antibody preparations and HIV vaccines to the mother and/or child. A number of animal studies using active and passive products have shown efficacy in similar settings, providing hope that a similar strategy will work in humans. Clinical trials conducted through the US National Institutes of Health AIDS Clinical Trials Programs have focused on such an approach. This article summarizes the results of a number of these trials, including AVEG (AIDS Vaccine Evaluation Group) 104, a phase I study of active immunization of HIV-infected pregnant women with an HIV gp120 subunit vaccine, and PACTG (Pediatrics AIDS Clinical Trial Group) 185, a phase III efficacy trial of HIV immunoglobulin administered to HIV-infected pregnant women and their newborns. A number of HIV vaccine trials have been performed in HIV-exposed and -infected children. These include PACTG 218, a vaccine immunotherapy phase I trial of HIV subunit vaccines administered to asymptomatic HIV-infected children, and two phase I vaccine trials in HIV-exposed children, PACTG 230 and 326. While the results of PACTG 230 were encouraging, the gp120 subunit vaccines were not capable of generating cytotoxic T-cell (CTL) responses. A subsequent phase I study, PACTG 326, utilized a canarypox vectored HIV vaccine (ALVAC vCP205, Sanofi Pasteur), which was previously shown to generate CTL responses in HIV-uninfected adults. The vaccines were safe but the immunogenicity was poor when compared with results of adult studies. Specifically, no antibody responses were found, lymphoproliferative responses to HIV-specific antigens were found in <50% of vaccinees, and CTL responses, modest in nature, were seen in approximately 50% of vaccinees.Future planned vaccine studies are focused on prime-boost approaches, using live recombinant vectors or DNA vaccines combined with subunit vaccines to stimulate both cellular and antibody responses. HIV vaccines may have special utility in newborns, in infants who continue to breastfeed, and in pre-adolescent children before they become sexually active. However, to date, candidate HIV vaccines capable of generating robust immunologic responses in these populations have been disappointing. PMID- 16220995 TI - Management options for bipolar disorder in children and adolescents. AB - During recent years there has been a dramatic increase in the use of psychotropic medication for the treatment of bipolar disorder (BPD) in children. There is an emerging set of data to support this use.Mood stabilizers, including lithium and valproic acid (valproate sodium), have generally formed the mainstay of treatment in children and adolescents with BPD. However, the atypical antipsychotics, such as risperidone, aripiprazole, and quetiapine may be more effective as first-line treatment options and in some ways easier to use than the traditional mood stabilizers. As in adults, mood stabilization is often difficult to achieve in pediatric patients with BPD, and combined treatment with mood stabilizers and atypical antipscyhotics is commonly used. Data from controlled trials of psychotropic medications in children and adolescents with BPD are very limited, and hence, in the majority of cases physicians base their treatment decisions on data from case reports, case series, or open trials. More controlled studies of both monotherapy and polypharmacotherapy for BPD in children and adolescents are needed. PMID- 16220998 TI - When clinical trials are compromised: a perspective from a patient advocate. PMID- 16220997 TI - Treatment of immune thrombocytopenic purpura in children : current concepts. AB - Treatment of immune thrombocytopenic purpura (ITP), the most common bleeding disorder of childhood, is a controversial subject for most practitioners. Diagnosis and management of ITP has historically been based primarily on expert opinion rather than on evidence. Due to a paucity of carefully conducted clinical trials in children, the management of ITP varies widely, ranging from observation only, to aggressive management with intravenous immunoglobulin (IVIG), intravenous anti-D rhesus (Rh)0 immunoglobulin (IV RhIG), corticosteroids, and splenectomy. To address the controversies, the American Society of Hematology (ASH) and the British Society for Hematology (BSH) have developed ITP practice guidelines. These guidelines, based on expert opinion, differ in their recommendations for treatment. The ASH guidelines favor therapy based on a low platelet count, and the more current BSH guidelines recommend a more conservative 'wait and watch' approach. In addition to treating children with severe bleeding symptoms, there is a tendency (not evidence based) to treat early in order to prevent a life-threatening bleeding episode, including intracerebral hemorrhage. Corticosteroids are a highly effective therapy, inexpensive, and can usually increase the platelet count within hours to days. However, chronic or prolonged use is associated with toxicity. In the US, based on the knowledge of known toxicities of corticosteroids, as well as the efficacy of alternative treatments (IV RhIG, IVIG), many pediatricians prefer to treat with IVIG and IV RhIG, reserving corticosteroid treatment for serious bleeding or refractory disease. However, in the UK, for the most part, corticosteroids are used as first-line therapy in children with ITP. Splenectomy is rarely indicated in children except for those with life-threatening bleeding and chronic, severe ITP with impairment of quality of life. For children who develop chronic or refractory ITP, immunosuppressive drugs and/or chemotherapy agents may offer some promise. However, the long-term effects of these drugs in children are unknown and they should not be considered unless there is unequivocal evidence that the patient is refractory to IV RhIG, IVIG, and corticosteroids. To date, virtually all of the randomized clinical trials conducted in children with ITP have focused on platelet counts as the sole outcome measure. Only carefully designed, multicenter, randomized clinical trials comparing the effects of different treatment modalities in terms of bleeding, quality of life, adverse effects, and treatment-related costs will be able to address the controversies surrounding childhood ITP treatment and allow management of this condition to be based on scientific data rather than treatment philosophy. PMID- 16220999 TI - Beyond trial registration: a global trial bank for clinical trial reporting. PMID- 16221000 TI - Clinical trial registration: the differing views of industry, the WHO, and the Ottawa Group. PMID- 16221002 TI - Psychotherapy, classism, and the poor: conspicuous by their absence. AB - Four decades of research have delineated the need for improved psychotherapeutic opportunities for poor clients, yet psychotherapists remain contradictory in their stance regarding service to the poor. Despite periodic calls within the field to address the needs of poor people, evidence from the psychotherapeutic literature suggests that the poor are still largely absent from consideration. What barriers prevent psychotherapists from enacting their professional principles more consistently on behalf of poor clients? The author suggests that unexamined classist assumptions constitute a significant obstacle for practitioners and presents the experience of confronting her own classism to illustrate the operation of these attitudinal barriers. PMID- 16221001 TI - Positive affect and the complex dynamics of human flourishing. AB - Extending B. L. Fredrickson's (1998) broaden-and-build theory of positive emotions and M. Losada's (1999) nonlinear dynamics model of team performance, the authors predict that a ratio of positive to negative affect at or above 2.9 will characterize individuals in flourishing mental health. Participants (N=188) completed an initial survey to identify flourishing mental health and then provided daily reports of experienced positive and negative emotions over 28 days. Results showed that the mean ratio of positive to negative affect was above 2.9 for individuals classified as flourishing and below that threshold for those not flourishing. Together with other evidence, these findings suggest that a set of general mathematical principles may describe the relations between positive affect and human flourishing. PMID- 16221003 TI - Beyond the optimal contact strategy: a reality check for the contact hypothesis. AB - The contact hypothesis proposes that interaction between members of different groups reduces intergroup prejudice if--and only if--certain optimal conditions are present. For over 50 years, research using this framework has explored the boundary conditions for ideal contact and has guided interventions to promote desegregation. Although supporting the contact hypothesis in principle, the authors critique some research practices that have come to dominate the field: (a) the prioritizing of the study of interactions occurring under rarefied conditions, (b) the reformulation of lay understandings of contact in terms of a generic typology of ideal dimensions, and (c) the use of shifts in personal prejudice as the primary measure of outcome. The authors argue that these practices have limited the contact hypothesis both as an explanation of the intergroup dynamics of desegregation and as a framework for promoting social psychological change. In so arguing, the authors look toward a complementary program of research on contact and desegregation. PMID- 16220996 TI - Osteoporosis in children and adolescents: etiology and management. AB - Bone mass increases progressively during childhood, but mainly during adolescence when approximately 40% of total bone mass is accumulated. Peak bone mass is reached in late adolescence, and is a well recognised risk factor for osteoporosis later in life. Thus, increasing peak bone mass can prevent osteoporosis. The critical interpretation of bone mass measurements is a crucial factor for the diagnosis of osteopenia/osteoporosis in children and adolescents. To date, there are insufficient data to formally define osteopenia/osteoporosis in this patient group, and the guidelines used for adult patients are not applicable. In males and females aged <20 years the terminology 'low bone density for chronologic age' may be used if the Z-score is less than -2. For children and adolescents, this terminology is more appropriate than osteopenia/osteoporosis. Moreover, the T-score should not be used in children and adolescents. Many disorders, by various mechanisms, may affect the acquisition of bone mass during childhood and adolescence. Indeed, the number of disorders that have been identified as affecting bone mass in this age group is increasing as a consequence of the wide use of bone mass measurements. The increased survival of children and adolescents with chronic diseases or malignancies, as well as the use of some treatment regimens has resulted in an increase in the incidence of reduced bone mass in this age group. Experience in treating the various disorders associated with osteoporosis in childhood is limited at present. The first approach to osteoporosis management in children and adolescents should be aimed at treating the underlying disease. The use of bisphosphonates in children and adolescents with osteoporosis is increasing and their positive effect in improving bone mineral density is encouraging. Osteoporosis prevention is a key factor and it should begin in childhood. Pediatricians should have a fundamental role in the prevention of osteoporosis, suggesting strategies to achieve an optimal peak bone mass. PMID- 16221004 TI - Global mobility for psychologists: the role of psychology organizations in the United States, Canada, Europe, and other regions. AB - Global mobility for psychologists is rapidly improving because of an emerging consensus on recognition standards, the demand for cross-border mobility both internal and external to the profession, and the efforts of membership, credentialing, and regional organizations to promote mobility. In the United States, multiple credentialing organizations promote mobility, primarily through individual endorsement of credentials. The Canadian regulatory boards signed a mutual recognition agreement implementing fast-track mechanisms for licensed psychologists seeking mobility and a competency-based assessment for initial registration. Europe plans to reduce barriers to mobility through mutual recognition of qualifications via a EuroPsy diploma that provides a benchmark for professional psychology education and training. Other regions have not made as much progress with regard to promoting mobility, as many do not yet even regulate the practice of psychology. The authors examine who seeks mobility, which geographic regions promote mobility, and by which mechanisms. PMID- 16221005 TI - Jerome D. Frank (1910-2005). PMID- 16221006 TI - Frank A. Logan (1924-2004). PMID- 16221007 TI - Kenneth Lucien Dion (1944-2004). PMID- 16221008 TI - Melvin H. Marx (1919-2005). PMID- 16221009 TI - Nursing and psychological treatments. AB - Presents a comment on "Psychological Treatments" by D. H. Barlow. In his article, Barlow pointed to the need "to solidify the identification of psychology as a health care profession" by changing the terminology of practice in the health care context from psychotherapy to psychological treatments and suggested that the only persons qualified to carry out such interventions are doctoral-level psychologists. Unfortunately, there was no discussion of the health care professionals who already provide psychological treatments in health care settings and their contribution to the evidence base supporting such treatment. The authors find several aspects of the article to be problematic. Overall, the authors feel that suggesting that psychology should claim treatment of psychological disorders and psychological components of physical disorders in health care settings as exclusively its own domain ignores the research and clinical contributions of others. PMID- 16221010 TI - "Psychotherapy" versus "treatment". AB - Presents a comment on "Psychological treatments" by D. H. Barlow. Barlow proposed that we distinguish between the terms "treatment" and "psychotherapy." The author believes that not only is the distinction unnecessary, but that its implications could have negative consequences for the field of clinical psychology. It is the proposed distinguishing feature that treatments are "specifically tailored to the pathological process that is causing the impairment and distress" that is most problematic. Clinical psychology does not need a distinction that further exacerbates the split between researchers and practitioners. PMID- 16221011 TI - Exploring unique roles for psychologists. AB - Presents a comment on "Psychological Treatments" by D. H. Barlow. Barlow highlighted unique roles that psychologists can play in mental health service delivery by providing psychological treatments--treatments that psychologists would be uniquely qualified to design and deliver. In support of Barlow's position, the authors draw from their own clinical practice with special psychiatric populations, such as adults with severe and persistent mental illness and behaviorally disordered youths, to illustrate some potential unique roles for psychologists. The authors believe psychologists are uniquely trained to design such individualized functional behavioral analysis protocols because of their training in research design, behavior analysis, learning theory, and behavior change. Psychologists may also be uniquely qualified to design, implement, and evaluate many specialized therapy techniques, as Barlow has outlined and suggested. PMID- 16221013 TI - Still looking for Poppa. AB - In 1992, V. Phares published an article titled "Where's Poppa?: The Relative Lack of Attention to the Role of Fathers in Child and Adolescent Psychopathology". Since that time, there have been modest gains in the research literature on clinical child issues, but there remains a wide gap between the inclusion of mothers and fathers in clinical child and family research. To provide an update of this issue for the field of developmental psychopathology, the authors of this comment conducted an updated review and analysis of the research on fathers and developmental psychopathology. These current data were compared with the data from the Phares and Compas (1992) study. It was found that there continues to be a dearth of research on fathers and developmental psychopathology. PMID- 16221014 TI - Genetic effects on women's positive mental health: do marital relationships and social support matter? AB - Interpersonal relationships are important factors in mental health. A genetically sensitive design was used to examine associations among marital quality, adequacy of social support, and 2 aspects of positive mental health in a sample of 652 Swedish twin women and their families. There were 3 main findings. First, the covariance between relationships and positive mental health was partially accounted for by common genetic variance. Second, nonshared environmental influences played a substantial role in the covariance among the 3 constructs, with evidence for husbands being a source of this influence. Finally, different patterns of associations were found between relationships and 2 aspects of mental health, well-being and global self-worth, which shows how seemingly similar constructs can be differentially associated with relationships. Together, these findings emphasize the importance of genetically informed studies in family research and the role of the environment and interpersonal relationships in promoting and improving mental health. PMID- 16221015 TI - Incarcerated mothers' contact with children, perceived family relationships, and depressive symptoms. AB - Concurrent relations among contact with children, perceived family relationships, early experiences of relationship disconnection and trauma, and maternal depressive symptoms were examined in 94 incarcerated mothers with children between the ages of 2 and 7 years. Qualitative analysis revealed that most mothers experienced intense distress when initially separated from their children, although many women currently viewed the situation in a more balanced way. Quantitative findings indicated that fewer visits from children and early experiences of relationship disconnection and trauma were associated with elevated maternal depressive symptoms. Mother-child relationships were more positive when mothers had more frequent telephone contact with older children. Moreover, conflicted relationships with caregivers related to less contact between mothers and their children. Results highlight the need for mental health services for incarcerated women and suggest that interventions aimed at increasing contact between imprisoned mothers and their children should consider the quality of the mother-caregiver relationship. PMID- 16221016 TI - The intergenerational transmission of perfectionism: parents' psychological control as an intervening variable. AB - The present study investigated the role of parental (adaptive and maladaptive) intrapersonal perfectionism as a predictor of parental psychological control and the role of parents' psychological control in the intergenerational transmission of perfectionism in a sample of female late adolescents and their parents. First, parental maladaptive perfectionism, but not parental adaptive perfectionism, significantly predicted parents' psychological control even when controlling for parents' neuroticism. This relationship was found to be stronger for fathers than for mothers. Second, a significant direct relationship was found between mothers' and daughters' maladaptive perfectionism but not between fathers' and daughters' maladaptive perfectionism. Third, process analyses showed that, for both mothers and fathers, psychological control is an intervening variable in the relationship between parents' and daughters' maladaptive perfectionism. PMID- 16221017 TI - The acculturation gap-distress hypothesis among high-risk Mexican American families. AB - The authors tested the acculturation gap-distress hypothesis by examining whether parent-adolescent acculturation gaps were associated with greater conflict and youth conduct problems among 260 high-risk Mexican American families. The authors operationalized acculturation gaps in 2 ways: parent-youth mismatches in acculturation style, and parent-youth discrepancies in acculturation toward both mainstream and heritage cultures. Acculturation gaps were common, but results of hierarchical regression analyses indicated that parent-youth discrepancies in acculturation toward mainstream and heritage cultures were not related to increased conflict or youth conduct problems. Conduct problems were no higher in families in which the adolescent was more aligned with mainstream culture than the parent. Unexpectedly, the authors found more youth conduct problems in families in which the youth was more aligned with traditional culture than the parent. The results call into question the assumption that the more rapid acculturation of adolescents to American culture inevitably leads to distress in minority families. PMID- 16221018 TI - In the eyes of the beholder: cognitive appraisals as mediators of the association between interparental conflict and youth maladjustment. AB - Although the association between interparental conflict and youth maladjustment has been established, processes underlying this relationship are less understood. In this investigation, children's conflict appraisals were examined as mediating variables. In Study 1, 1,893 6th graders reported their perceptions of conflict and appraisals of threat and self-blame. Youth and teachers reported on externalizing and internalizing problems. In Study 2, 416 married parents from the larger sample reported their conflict and youth maladjustment. Children's appraisals of coping efficacy also were examined. Perceived threat, self-blame, and coping efficacy were salient mediators of overt conflict and triangulation, particularly for internalizing problems. Findings indicate that children's beliefs about interparental conflict play an important role in their adjustment to this family stressor. PMID- 16221019 TI - Assessing how much couples work at their relationship: the behavioral self regulation for effective relationships scale. AB - It is widely believed that satisfying couple relationships require work by the partners. The authors equated the concept of work to relationship self-regulation and developed a scale to assess this construct. A factor analysis of the scale in a sample of 187 newlywed couples showed it comprised 2 factors of relationship strategies and effort. The factor structure was replicated in an independent sample of 97 newlywed couples. In both samples the scale had good internal consistency and high convergent validity between self- and partner-report forms. Self-regulation accounted for substantial variance in relationship satisfaction in both newlywed samples and in a 3rd sample of 61 long-married couples. The self regulation and satisfaction association was independent of mood or self-report common method variance. PMID- 16221021 TI - Parental illness, family functioning, and adolescent well-being: a family ecology framework to guide research. AB - A family ecology model for understanding adolescents' reactions to parental illness is presented and used to (a) critically evaluate existing research that examines direct effects of parental illness on family functioning and youth well being and (b) provide a blueprint for future research in the area. Theoretical, clinical, and empirical literature is reviewed for each mediational and moderational pathway in the model, and limitations of the existing research are discussed. The blueprint for future research emphasizes a greater understanding of the mediational pathways in the model, which is essential for developing effective interventions for families experiencing parental illness. In addition, greater elucidation of moderator variables, such as the youth's developmental stage, social support, and cultural norms will provide critical information on contextual factors that enhance or impede adolescents' adaptation to serious parental illness. PMID- 16221020 TI - Family functioning and children's adjustment: associations among parents' depressed mood, marital hostility, parent-child hostility, and children's adjustment. AB - Relations between parents' depressed mood, marital conflict, parent-child hostility, and children's adjustment were examined in a community sample of 136 ten-year-olds and their parents. Videotaped observational and self-report data were used to examine these relations in path analyses. A proposed model was tested in which mothers' and fathers' depressed mood and marital hostility were associated with children's adjustment problems through disruptions in parent child relationships. Results showed that both mothers' and fathers' marital hostility were linked to parent-child hostility, which in turn was linked to children's internalizing problems. Fathers' depressed mood was linked to children's internalizing problems indirectly through father-child hostility. Fathers' depressed mood was directly linked to children's externalizing problems and indirectly linked through father-child hostility. For mothers, marital hostility was directly linked to children's externalizing problems, and marital hostility in fathers was indirectly linked to children's externalizing problems through father-child hostility. PMID- 16221022 TI - Adult attachment and marital interaction as predictors of whole family interactions during the transition to parenthood. AB - This study examined the role of parental adult attachment and couples' prenatal and postnatal marital interactions in predicting the quality of family interactions 24 months after the birth of the couple's first child. Father's prenatal marital withdrawal and mother's postnatal marital withdrawal were associated with less adaptive family interactions at 24 months. Families with fathers who had an insecure attachment, as assessed by the Adult Attachment Interview, showed less positive and more negative interactions at 24 months, but only when there were higher levels of negative escalation in the couple's marriage prenatally. Findings are discussed in terms of the importance of exploring the transition to parenthood at the broader family level. PMID- 16221023 TI - Patterns of posttraumatic stress symptoms in parents of childhood cancer survivors. AB - Posttraumatic stress (PTS) symptoms have been reported in mothers and fathers of childhood cancer survivors; however, little is known about patterns of PTS in these families. Cluster analysis was applied to the Posttraumatic Stress Disorder (PTSD) Reaction Index scores of 98 couples parenting adolescent childhood cancer survivors to describe patterns of PTS in families, yielding the following 5 clusters: Minimal PTS, Mothers Elevated, Disengaged, Fathers Elevated, and Elevated PTS. The clusters were validated using data from a structured psychiatric interview, an additional self-report measure of PTS, and an index of family functioning. These clinically meaningful patterns reveal that a majority of families had at least one parent with moderate to severe PTS, which supports development of family-based interventions for this population. PMID- 16221024 TI - Measurement equivalence and differential item functioning in family psychology. AB - Several hypotheses in family psychology involve comparisons of sociocultural groups. Yet the potential for cross-cultural inequivalence in widely used psychological measurement instruments threatens the validity of inferences about group differences. Methods for dealing with these issues have been developed via the framework of item response theory. These methods deal with an important type of measurement inequivalence, called differential item functioning (DIF). The authors introduce DIF analytic methods, linking them to a well-established framework for conceptualizing cross-cultural measurement equivalence in psychology (C.H. Hui and H.C. Triandis, 1985). They illustrate the use of DIF methods using data from the Project on Human Development in Chicago Neighborhoods (PHDCN). Focusing on the Caregiver Warmth and Environmental Organization scales from the PHDCN's adaptation of the Home Observation for Measurement of the Environment Inventory, the authors obtain results that exemplify the range of outcomes that may result when these methods are applied to psychological measurement instruments. PMID- 16221025 TI - Do street youths' perceptions of their caregivers predict HIV-risk behavior? AB - This study examined street youths' perceptions of their caregivers and the association between these perceptions and HIV-risk behavior in a random probability sample of 715 12- to 23-year-old street youths from Los Angeles and San Diego, CA (mean age, 18.7 years). All participants had been homeless at some point during the past 12 months, with 70% recruited from nonshelter sites. Although youths reported high rates of hostility, unavailability, substance use, and legal problems among their caregivers, 86% reported that their caregivers had at least one attribute associated with support. Caregiver problems were associated with youth having had more sexual partners in the past 30 days and having higher risk drug use. High caregiver support was associated with more sexual partners and lower use of condoms with steady partners. Caregiver attributes did not predict condom use with transient partners. PMID- 16221026 TI - Dyadic adjustment in chronic illness: does relationship talk matter? AB - Relationship talk involves talking about the nature and state of one's relationship. To determine the effectiveness of talking about the relationship when one spouse has a chronic illness, the study involved completion of a confidential questionnaire by 182 married couples. Ninety of these were couples in which both partners were healthy, and 92 were couples in which one spouse had a chronic illness. Results of multilevel modeling analyses showed that the association between relationship talk and dyadic adjustment was stronger for women than for men and for couples with an ill spouse than for couples where both spouses were healthy. These findings highlight the importance of taking a relationship perspective and suggest that relationship talk is a potentially useful tool couples can use in their repertoire of relationship-enhancing behaviors during chronic illness. PMID- 16221028 TI - People are variables too: multilevel structural equations modeling. AB - The article uses confirmatory factor analysis (CFA) as a template to explain didactically multilevel structural equation models (ML-SEM) and to demonstrate the equivalence of general mixed-effects models and ML-SEM. An intuitively appealing graphical representation of complex ML-SEMs is introduced that succinctly describes the underlying model and its assumptions. The use of definition variables (i.e., observed variables used to fix model parameters to individual specific data values) is extended to the case of ML-SEMs for clustered data with random slopes. Empirical examples of multilevel CFA and ML-SEM with random slopes are provided along with scripts for fitting such models in SAS Proc Mixed, Mplus, and Mx. Methodological issues regarding estimation of complex ML SEMs and the evaluation of model fit are discussed. Further potential applications of ML-SEMs are explored. PMID- 16221027 TI - Infidelity in couples seeking marital therapy. AB - The revelation of an affair is often an emotionally explosive event for a couple, yet little is known about specific individual and relationship factors that accompany infidelity. The present study examined the qualities of individuals and couples that differentiate couples with (n = 19) and without (n = 115) infidelity using couples from a randomized clinical trial of marital therapy. Findings indicated that couples with infidelity showed greater marital instability, dishonesty, arguments about trust, narcissism, and time spent apart. Gender also proved to be a significant moderator of several effects. Men who had participated in affairs showed increased substance use, were older, and were more sexually dissatisfied. Results offer initial clues to concomitants of affairs for couple therapists. PMID- 16221029 TI - Structural equation modeling of paired-comparison and ranking data. AB - L. L. Thurstone's (1927) model provides a powerful framework for modeling individual differences in choice behavior. An overview of Thurstonian models for comparative data is provided, including the classical Case V and Case III models as well as more general choice models with unrestricted and factor-analytic covariance structures. A flow chart summarizes the model selection process. The authors show how to embed these models within a more familiar structural equation modeling (SEM) framework. The different special cases of Thurstone's model can be estimated with a popular SEM statistical package, including factor analysis models for paired comparisons and rankings. Only minor modifications are needed to accommodate both types of data. As a result, complex models for comparative judgments can be both estimated and tested efficiently. PMID- 16221030 TI - The role of nonlinear factor-to-indicator relationships in tests of measurement equivalence. AB - Measurement invariance is a necessary condition for the evaluation of factor mean differences over groups or time. This article considers the potential problems that can arise for tests of measurement invariance when the true factor-to indicator relationship is nonlinear (quadratic) and invariant but the linear factor model is nevertheless applied. The factor loadings and indicator intercepts of the linear model will diverge across groups as the factor mean difference increases. Power analyses show that even apparently small quadratic effects can result in rejection of measurement invariance at moderate sample sizes when the factor mean difference is medium to large. Recommendations include the identification of nonlinear relationships using diagnostic plots and consideration of newly developed methods for fitting nonlinear factor models. PMID- 16221031 TI - An alternative to Cohen's standardized mean difference effect size: a robust parameter and confidence interval in the two independent groups case. AB - The authors argue that a robust version of Cohen's effect size constructed by replacing population means with 20% trimmed means and the population standard deviation with the square root of a 20% Winsorized variance is a better measure of population separation than is Cohen's effect size. The authors investigated coverage probability for confidence intervals for the new effect size measure. The confidence intervals were constructed by using the noncentral t distribution and the percentile bootstrap. Over the range of distributions and effect sizes investigated in the study, coverage probability was better for the percentile bootstrap confidence interval. PMID- 16221032 TI - Omnibus hypothesis testing in dominance-based ordinal multiple regression. AB - Often quantitative data in the social sciences have only ordinal justification. Problems of interpretation can arise when least squares multiple regression (LSMR) is used with ordinal data. Two ordinal alternatives are discussed, dominance-based ordinal multiple regression (DOMR) and proportional odds multiple regression. The Q2 statistic is introduced for testing the omnibus null hypothesis in DOMR. A simulation study is discussed that examines the actual Type I error rate and power of Q2 in comparison to the LSMR omnibus F test under normality and non-normality. Results suggest that Q2 has favorable sampling properties as long as the sample size-to-predictors ratio is not too small, and Q2 can be a good alternative to the omnibus F test when the response variable is non-normal. PMID- 16221033 TI - Studentized maximum root procedures for coherent analyses of two-factor fixed effects designs. AB - The authors provide generalizations of R. J. Boik's (1993) studentized maximum root (SMR) procedure that allow for simultaneous inference on families of product contrasts including simple effect contrasts and differences among simple effect contrasts in coherent analyses of data from 2-factor fixed-effects designs. Unlike the F-based simultaneous test procedures (STPs) proposed by M. A. Betz and K. R. Gabriel (1978) for coherent analyses allowing for inferences on all factorial contrasts, SMR STPs are designed for analyses where each contrast of interest is a product contrast, which is usually the case in analyses of data from factorial experiments. When both factors have more than 2 levels, SMR STPs always provide more power and precision than F STPs for inferences on product contrasts. PMID- 16221034 TI - Building face composites can harm lineup identification performance. AB - Face composite programs permit eyewitnesses to build likenesses of target faces by selecting facial features and combining them into an intact face. Research has shown that these composites are generally poor likenesses of the target face. Two experiments tested the proposition that this composite-building process could harm the builder's memory for the face. In Experiment 1 (n = 150), the authors used 50 different faces and found that the building of a composite reduced the chances that the person could later identify the original face from a lineup when compared with no composite control conditions or with yoked composite-exposure control conditions. In Experiment 2 (n = 200), the authors found that this effect generalized to a simulated-crime video, but mistaken identifications from target absent lineups were not inflated by composite building. PMID- 16221035 TI - Assessing the effectiveness of various auditory cues in capturing a driver's visual attention. AB - This study was designed to assess the potential benefits of using spatial auditory warning signals in a simulated driving task. In particular, the authors assessed the possible facilitation of responses (braking or accelerating) to potential emergency driving situations (the rapid approach of a car from the front or from behind) seen through the windshield or the rearview mirror. Across 5 experiments, the authors assessed the efficacy of nonspatial-nonpredictive (neutral), spatially nonpredictive (50% valid), and spatially predictive (80% valid) car horn sounds, as well as symbolic predictive and spatially presented symbolic predictive verbal cues (the words "front" or "back") in directing the participant's visual attention to the relevant direction. The results suggest that spatially predictive semantically meaningful auditory warning signals may provide a particularly effective means of capturing attention. PMID- 16221036 TI - Influence of virtual reality training on the roadside crossing judgments of child pedestrians. AB - The roadside crossing judgments of children aged 7, 9, and 11 years were assessed relative to controls before and after training with a computer-simulated traffic environment. Trained children crossed more quickly, and their estimated crossing times became better aligned with actual crossing times. They crossed more promptly, missed fewer safe opportunities to cross, accepted smaller traffic gaps without increasing the number of risky crossings, and showed better conceptual understanding of the factors to be considered when making crossing judgments. All age groups improved to the same extent, and there was no deterioration when children were retested 8 months later. The results are discussed in relation to theoretical arguments concerning the extent to which children's pedestrian judgments are amenable to training. PMID- 16221037 TI - Influence of illness script components and medical practice on medical decision making. AB - Illness scripts are knowledge structures composed of consequences, enabling conditions, and faults. The effects of illness script components--consequences and enabling conditions--and physician factors on referral decisions for gastrointestinal disorders were investigated. The hypothesis that consequences and enabling conditions increase the likelihood of referral was confirmed and several interactions between consequences and enabling conditions were found. The hypothesis that physician factors moderate the effect of enabling conditions was also confirmed, but (contrary to illness script theory) evidence was also found for moderation of consequences. Both enabling conditions and consequences were found to be moderated by physician factors to a larger extent than previously assumed by illness script theory. PMID- 16221039 TI - Golden cows and sacred geese: it's all Greek to me. PMID- 16221038 TI - Social recognition memory: the effect of other people's responses for previously seen and unseen items. AB - When people discuss their memories, what one person says can influence what another person reports. In 3 studies, participants were shown sets of stimuli and then given recognition memory tests to measure the effect of one person's response on another's. The 1st study (n = 24) used word recognition with participant-confederate pairs and found that the effect of confederate responses on participant responses was larger for previously unseen items than for previously seen items (omega(p) = .23). This finding was replicated in the 2nd study, which used photographs of cars (n = 24). In the 3rd study (n = 54), which used photographs of faces with participant pairs, the effect was also larger for unseen items. Results indicate that people rely more on other people's memories for unremembered objects than for remembered objects. This is important for both theories of memory and applications (e.g., witnesses talking, students studying together). PMID- 16221040 TI - An overall strategy for the testing of chemicals for human hazard and risk assessment under the EU REACH system. AB - In its White Paper, "Strategy for a Future Chemicals Policy," published in 2001, the European Commission (EC) proposed the REACH (Registration, Evaluation and Authorisation of CHemicals) system to deal with both existing and new chemical substances. This system is based on a top-down approach to toxicity testing, in which the degree of toxicity information required is dictated primarily by production volume (tonnage). If testing is to be based on traditional methods, very large numbers of laboratory animals could be needed in response to the REACH system, causing ethical, scientific and logistical problems that would be incompatible with the time-schedule envisaged for testing. The EC has emphasised the need to minimise animal use, but has failed to produce a comprehensive strategy for doing so. The present document provides an overall scheme for predictive toxicity testing, whereby the non-animal methods identified and discussed in a recent and comprehensive ECVAM document, could be used in a tiered approach to provide a rapid and scientifically justified basis for the risk assessment of chemicals for their toxic effects in humans. The scheme starts with a preliminary risk assessment process (involving available information on hazard and exposure), followed by testing, based on physicochemical properties and (Q)SAR approaches. (Q)SAR analyses are used in conjunction with expert system and biokinetic modelling, and information on metabolism and identification of the principal metabolites in humans. The resulting information is then combined with production levels and patterns of use to assess potential human exposure. The nature and extent of any further testing should be based strictly on the need to fill essential information gaps in order to generate adequate risk assessments, and should rely on non-animal methods, as far as possible. The scheme also includes a feedback loop, so that new information is used to improve the predictivity of computational expert systems. Several recommendations are made, the most important of which is that the European Union (EU) should actively promote the improvement and validation of (Q)SAR models and expert systems, and computer-based methods for biokinetic modelling, since these offer the most realistic and most economical solution to the need to test large numbers of chemicals. PMID- 16221041 TI - Heterologous expression of mouse cytochrome P450 2e1 in V79 cells: construction and characterisation of the cell line and comparison with V79 cell lines stably expressing rat P450 2E1 and human P450 2E1. AB - A V79 Chinese hamster cell line was constructed for stable expression of mouse cytochrome P450 2e1 (Cyp2e1), as an addition to the existing cell battery consisting of cell lines stably expressing rat CYP2E1 and human CYP2E1 (V79 Cell Battery). The aim was to establish a cell battery that offers the in vitro possibility of investigating species-specific differences in the toxicity and metabolism of chemicals representing substrates for CYP2E1. The newly established cell line (V79m2E1) effectively expressed Cyp2e1 in the catalytically active form. The expression of catalytically active CYP2E1 in V79m2E1 cells was maintained over several months in culture, as demonstrated by Western Blotting and chlorzoxazone (CLX) 6-hydroxylase activity. The cells exhibited CLX 6 hydroxylase activity with a Km of 27.8 microM/l and Vmax of 40 pmol/mg protein/minute, compared with a Km of 28.2/28.6 microM/l and a Vmax of 130/60 pmol/mg protein/minute from V79r2E1/V79h2E1 cells. Furthermore, the CYP2E1 dependent mutagenicity of N-nitrosodimethylamine could be demonstrated in the V79m2E1 cells. Therefore, the new cell battery permits the interspecies comparison of CYP2E1-dependent toxicity and of metabolism of chemicals between humans and the two major rodent species--the rat and the mouse--that are usually used in classical toxicity studies. PMID- 16221042 TI - Using in vitro prediction models instead of the rabbit eye irritation test to classify and label new chemicals: a post hoc data analysis of the international EC/HO validation study. AB - The international validation study on alternative methods to replace the Draize rabbit eye irritation test, funded by the European Commission (EC) and the British Home Office (HO), took place during 1992-1994, and the results were published in 1995. The results of this EC/HO study are analysed by employing discriminant analysis, taking into account the classification of the in vivo data into eye irritation classes A (risk of serious damage to eyes), B (irritating to eyes) and NI (non-irritant). A data set for 59 test items was analysed, together with three subsets: surfactants, water-soluble chemicals, and water-insoluble chemicals. The new statistical methods of feature selection and estimation of the discriminant functions classification error were used. Normal distributed random numbers were added to the mean values of each in vitro endpoint, depending on the observed standard deviations. Thereafter, the reclassification error of the random observations was estimated by applying the fixed function of the mean values. Moreover, the leaving-one-out cross-classification method was applied to this random data set. Subsequently, random data were generated r times (for example, r = 1000) for a feature combination. Eighteen features were investigated in nine in vitro test systems to predict the effects of a chemical in the rabbit eye. 72.5% of the chemicals in the undivided sample were correctly classified when applying the in vitro endpoints lgNRU of the neutral red uptake test and lgBCOPo5 of the bovine opacity and permeability test. The accuracy increased to 80.9% when six in vitro features were used, and the sample was subdivided. The subset of surfactants was correctly classified in more than 90% of cases, which is an excellent performance. PMID- 16221043 TI - Comments on the House of Lords Select Committee proposal for the establishment of a Centre for the Three Rs. PMID- 16221044 TI - The ODAC Chronicles: part 6b. ODAC's structure and function. PMID- 16221046 TI - Thalidomide and dexamethasone: therapy for multiple myeloma. AB - Multiple myeloma is a plasma cell malignancy that remains incurable with current treatment approaches including high-dose therapy and autologous stem cell transplantation. Thalidomide represents a major advance in the treatment of this disorder, having demonstrated significant activity in all phases of the disease. Thalidomide exerts its antimyeloma effect through multiple mechanisms including antiangiogenesis, immunomodulation and induction of apoptosis in tumor cells, as well as its effect on the tumor microenvironment. Corticosteroids have formed the mainstay of myeloma therapy for decades along with the alkylating agents and have demonstrated synergy when used in combination with thalidomide. The combination of thalidomide and dexamethasone has demonstrated remarkable activity in the treatment of both newly diagnosed as well as relapsed myeloma, and has become an important addition to the armamentarium of myeloma therapies. Overall responses of approximately 70% have been seen with this combination in patients with newly diagnosed myeloma. The combination is associated with an increased risk of deep vein thrombosis necessitating routine prophylactic anticoagulation. Other drugs have been added to this combination that also result in improved response rates. Currently, this combination is used in newly diagnosed patients as an induction therapy prior to stem cell transplant, for those who fail to achieve adequate response to dexamethasone alone or in whom a relatively rapid response is desired based on clinical presentation. Thalidomide analogs with a better safety profile are currently undergoing evaluation in the clinic. PMID- 16221048 TI - Immunotherapy for human glioma: innovative approaches and recent results. AB - The outcome for malignant glioma patients remains dismal despite treatment with surgical resection, radiation and chemotherapy. The goal of immunotherapy is to eradicate or suppress the residual infiltrative component of these tumors. Although there is clinical evidence for cell-mediated antiglioma activity, there are special considerations that need to be accounted for in the design of immunotherapeutics for CNS tumors, such as possible differences in antigen presenting cells, trafficking of effector T-cells and immunosuppression. Previously characterized immunosuppression in glioma patients has included low peripheral blood lymphocyte counts, reduced delayed type hypersensitivity reactions to recall antigens, impaired mitogen-induced blastogenic responses by peripheral blood mononuclear cells, increased CD8+ suppressor T-cells, decreased CD4+ T-cell activity in vitro, diminished immunoglobulin synthesis by B-cells and impaired transmembrane signaling through the T-cell receptor/CD3 complex. Recent impairments that are being identified include anergy, failure of costimulation, lack of sufficient numbers of functional effector T-cells and the presence of T suppressor cells within the tumor microenvironment. It is proposed that these inherent problems will need to be overcome in order for immunotherapies to realize their potential. Paradoxically, the efficacy of recent clinical immunotherapies for glioma patients appears equivalent to that seen in other cancer immunotherapeutic approaches. This review will provide an overview of the juxtaposition of the immune system and CNS, and will discuss the most recent and ongoing immunotherapeutic clinical trials that are demonstrating promising results. PMID- 16221047 TI - Erlotinib in the treatment of non-small cell lung cancer. AB - Inhibition of the epidermal growth factor receptor is one of the most promising novel therapeutic strategies to be used in the treatment of patients with non small cell lung cancer. A number of compounds that target the epidermal growth factor receptor are in an advanced stage of clinical development including both antibodies directed against the receptor and small molecule inhibitors of epidermal growth factor receptor tyrosine kinase activity. This drug profile focuses on the development of erlotinib, an orally available inhibitor of epidermal growth factor receptor tyrosine kinase. Results of clinical trials are reviewed, two trials of erlotinib in combination, one with paclitaxel and carboplatin, the other with gemcitabine and cisplatin, and the National Cancer Institute of Canada--Clinical Trials Group BR21, the first study to demonstrate a survival benefit for this class of compound in non-small cell lung cancer. The future role of erlotinib in the management of patients with non-small cell lung cancer is also discussed. PMID- 16221049 TI - Photodynamic therapy in dermatology: current concepts in the treatment of skin cancer. AB - Photodynamic therapy is a treatment modality that is developing rapidly and increasing in utilization within various medical specialties, including dermatology. This technique requires the presence of a photosensitizer, light energy and molecular oxygen to selectively destroy pathologic cells. A thorough understanding of photobiology and tissue optics is necessary to correctly and effectively utilize photodynamic therapy in dermatology. Photodynamic therapy has been approved by the US Food and Drug Administration to treat actinic keratoses. In Europe, photodynamic therapy is currently being used in the treatment of actinic keratoses and basal cell carcinoma. Other off-label uses of photodynamic therapy have included cutaneous lesions of Bowen's disease, psoriasis, cutaneous T-cell lymphoma and acne. Most recently, photodynamic therapy has been employed in photorejuvenation. The advantages of photodynamic therapy include the capacity for noninvasive targeted therapy via topical application of the drug and local irradiation of involved areas, as well as the ability to generate excellent cosmetic results with minimal discomfort. This review summarizes the fundamentals of photodynamic therapy and its role in the treatment of cutaneous disorders, particularly skin malignancies. PMID- 16221050 TI - Can diet prevent nonmelanoma skin cancer progression? AB - Nonmelanoma skin cancer is the most prevalent malignant disease among light skinned individuals in the USA, accounting for approximately 1 million new cases annually. Solar ultraviolet radiation is known to be the major cause of skin cancer. The idea that diet, particularly dietary fat, may play a role in modulating cancer incidence has been based largely upon indirect epidemiologic evidence. However, few studies have found correlations of dietary fat intake with skin cancer incidence. Nevertheless, a large body of evidence with experimental animals and one clinical intervention trial, the latter avoiding the pitfalls of many epidemiologic investigations, clearly demonstrate that the level of dietary fat intake can have a significant influence on the occurrence of skin cancer in individuals at high risk. PMID- 16221051 TI - Treatment options for brain metastases from melanoma. AB - Brain metastases are a common complication of metastatic malignant melanoma, conferring an exceedingly poor prognosis. Diagnosis of brain metastasis often has significant implications for duration and quality of life, and management can be difficult due to rapid progression of disease and resistance to conventional therapies. This review focuses primarily on the published evidence for treatment modalities for brain metastases from melanoma, emerging technologies and outlines future directions for research. In summary, external-beam radiation alone appears effective in palliating symptoms. Surgical management of solitary or acutely symptomatic lesions appears to alleviate symptoms and provide the possibility of local control of disease. Stereotactic radiosurgery is an increasingly utilized technique for patients with a limited number of metastases and presents a less invasive alternative to craniotomy. Chemotherapy alone is relatively ineffective, although combined chemotherapy with external-beam radiation is being investigated. Future directions include combined modality therapy, the incorporation of novel agents and careful consideration of the structure of clinical trials for this disease. PMID- 16221052 TI - Cutaneous malignant melanoma: facts about sunbeds and sunscreen. AB - The incidence of cutaneous malignant melanoma is still increasing in most light skinned populations. Sunscreen use has been proposed for the primary prevention of melanoma. However, sunscreen use may increase time spent in the sun when users are willing to acquire a tan or to stay in the sun for a long time, which may increase melanoma risk. When sun exposure is not associated with the desire to acquire a tan or stay in the sun for a long time, sunscreen use may prevent squamous cell skin carcinoma. Sun protection should give priority to clothing and sun exposure reduction. Over the last 20 years, tan acquisition through exposure to artificial sources of ultraviolet radiation has become frequent among fair skinned adolescents and young adults. There is accumulating evidence that sunbed use is associated with melanoma when started before approximately 30 years of age. PMID- 16221053 TI - Autologous hematopoietic stem cell transplantation for pediatric solid tumors. AB - While advances in the treatment of pediatric cancers have increased cure rates, children with metastatic or recurrent solid tumors have a dismal prognosis despite initial transient responses to therapy. Autologous hematopoietic stem cell transplantation takes advantage of the steep dose-response relationship observed with many chemotherapeutic agents. While clearly demonstrated to improve outcomes in patients with metastatic neuroblastoma, autologous hematopoietic stem cell transplantation is also frequently used to treat patients with other high risk diseases such as Ewing sarcoma, osteosarcoma, rhabdomyosarcoma, Wilms' tumor, retinoblastoma, germ cell tumors, lymphomas and brain tumors. Most published experience consists of retrospective, single-arm studies; randomized clinical trials are lacking, due in part to the rarity of pediatric cancers treatable by autologous hematopoietic stem cell transplantation. These published literature demonstrate that autologous hematopoietic stem cell transplantation results in most cases in equivalent or superior outcomes when compared with conventional therapies. However, patient heterogeneity, patient selection, graft characteristics and processing and the varied conditioning regimens are additional factors to consider. Since the inception of autologous hematopoietic stem cell transplantation, regimen-related toxicity has markedly decreased and the vast majority of treatment failures are now due to disease recurrence. Prospective clinical trials are needed to identify specific high-risk patient populations, with randomization (when possible) to compare outcomes of patients undergoing autologous hematopoietic stem cell transplantation with those receiving standard therapy. In addition, investigators need to better define the role of autologous hematopoietic stem cell transplantation in these solid tumors, particularly in combination with other therapeutic modalities such as immunotherapy and novel cell processing methodologies. PMID- 16221054 TI - Lycopene and prostate cancer: emerging evidence. AB - Prostate cancer has the third highest incidence of all cancers in men worldwide and is the most common neoplasm diagnosed among men beyond middle age in many developed countries. Mounting evidence surrounding the consumption of tomato products has shown promise for the prevention of prostate cancer. This protective effect has more recently been linked to lycopene, the most abundant carotenoid in tomatoes. Lycopene is a natural pigment that gives the red color to many foods. In Western countries, 85% of dietary lycopene can be attributed to the consumption of tomato-based products. This article reviews emerging evidence from epidemiologic studies for the role of lycopene in prostate cancer prevention. The majority of evidence currently comes from observational studies, but recent human clinical trials and animal studies have provided additional support. Growing evidence on the biologic mechanisms of lycopene in prostate cancer prevention also confirm the epidemiologic findings. PMID- 16221055 TI - Investigational agents for epithelial ovarian cancer. AB - Ovarian cancer is the leading cause of gynecologic cancer deaths and accounts for 4% of women's cancer diagnoses and 5% of all cancer mortalities. Despite the ability of current chemotherapy and cytoreductive surgery to put patients in remission, most patients with advanced cancer will eventually relapse. Many advances in the treatment of ovarian cancer have been reported in the past several years and a historical background is provided. Attention will then turn to analogs of current chemotherapeutic agents, new cytotoxic drugs, targeted molecular therapy, intraperitoneal therapy and immunotherapy. This review will give a perspective on current drugs, potential agents and upcoming clinical trials. PMID- 16221056 TI - Postchemotherapy residual masses in advanced seminoma: current management and outcomes. AB - Although pure testicular seminoma is most often confined to the testis, it can present with advanced-stage bulky retroperitoneal metastases in nearly a quarter of cases. While highly treatable with cisplatin-based chemotherapy, up to 80% of patients with advanced disease are found to have a radiographically detectable residual mass after chemotherapy. The management of these postchemotherapy residual masses remains controversial. Surgical resection is technically challenging due to a desmoplastic reaction resulting from seminoma treatment and regression. In addition, these residual masses often demonstrate a protracted period of regression that can span several months to years. Surveillance protocols, therefore, may be appropriate for most patients. Several retrospective studies have supported surgical resection only for discrete, well-delineated masses over 3 cm in size. Despite the highly radiosensitive nature of seminoma, radiation therapy in this setting has not been shown to provide significant benefit, and may limit the tolerability of subsequent salvage chemotherapy. The incorporation of noninvasive imaging modalities, such as positron emission tomography, into the management algorithm may better delineate the presence of viable residual tumor and thus allow better risk stratification. PMID- 16221057 TI - Revisiting the role of hematopoietic stem cell transplantation in chronic lymphocytic leukemia. AB - Since the advent of hematopoietic stem cell transplantation more than 40 years ago, numerous methods of transplantation have been developed, modified and improved upon. Although hematopoietic stem cell transplantation has been used in a variety of malignant diseases since then, its use in the treatment of chronic lymphocytic leukemia has recently started to gain interest. Patients with chronic lymphocytic leukemia are generally elderly, and because of its relatively benign course, they were not considered suitable candidates for hematopoietic stem cell transplantation. Nonetheless, there have been marked improvements in transplantation techniques, including better conditioning regimens that have decreased treatment-related morbidity and mortality. In this article, the authors review the most recent data on hematopoietic stem cell transplantation in chronic lymphocytic leukemia as well as the change in risk stratification based on newer prognostic factors and its impact on treatment decisions in chronic lymphocytic leukemia. PMID- 16221058 TI - Imaging breast cancer response during neoadjuvant systemic therapy. AB - Neoadjuvant systemic therapy is used to enable breast-conserving surgery in patients with large primary operable breast cancers. It is important to be able to accurately assess response to systemic therapy, both to assist the surgeon and for prognostic purposes. Moreover, a proportion of women will fail to respond to treatment and would potentially benefit from either a change in therapy or earlier surgery rather than continuing completion of the planned course of treatment. Conventional techniques of assessing response (clinical examination, x ray mammography and breast ultrasound) rely on changes in tumor size, which are often delayed and do not always correlate with pathologic response. This review examines the evidence for functional imaging techniques including scintimammography, functional computed tomography, dynamic magnetic resonance imaging, spectroscopy and positron emission tomography. These techniques measure changes in tumor vasculature, metabolism or proliferation and may prove to be earlier and more sensitive measures of response to systemic therapy, thus enabling tailoring of an individual's treatment. PMID- 16221059 TI - Atypical teratoid rhabdoid tumors of childhood: diagnosis, treatment and challenges. AB - Atypical teratoid rhabdoid tumor of the brain was described as a unique entity in the late 1980s. It occurs primarily in early childhood but the true incidence of the disease is not yet known. At presentation, the differential diagnosis includes medulloblastoma, primitive neuroectodermal tumor, ependymoma and choroid plexus carcinoma. Atypical teratoid rhabdoid tumor behaves in a very aggressive manner and while cure is possible for a small minority of patients, no standard or effective therapy has been defined for most patients. Since its first description, considerable pathologic, cytogenetic and molecular characterizations, as described in this review, have been accomplished that provide insight into the possible molecular etiology of the disease and of malignant rhabdoid tumors that occur outside the CNS. Co-operative group clinical trials that focus solely on atypical teratoid rhabdoid tumor are needed that incorporate biologic studies along with evaluations of aggressive treatment approaches. The goal of these trials should be to increase the cure rate for children with atypical teratoid rhabdoid tumor and further increase our understanding not only of atypical teratoid rhabdoid tumor, but also of other pediatric brain tumors. PMID- 16221060 TI - Treatment of childhood acute myeloid leukemia. AB - Childhood acute myeloid leukemia is rare, but accounts for a significant number of malignancy-related deaths in this age group. However, the prognosis has improved over past decades, and survival rates of 60% and above have been reported. Still, this implies that more than a third of children and adolescents die from this disease. Moreover, treatment is intensive, and quality of life and late effects are worrying issues. Therefore, there is a need for further improved treatment of pediatric acute myeloid leukemia. This review describes several important developments in this respect, such as improved diagnostics, prognostic factors, subgroup-directed and tailored treatment, and targeted therapy. In addition, background information is provided and current treatment strategies are described, as well as the late effects of treatment. Most groups now have risk group adapted protocols, with allogeneic stem cell transplantation often being reserved for the higher risk group. Even in these cases, the benefit of stem cell transplantation has not been demonstrated beyond reasonable doubt with current high-intensive chemotherapy. Similarly, the use of cranial irradiation for CNS prophylaxis and maintenance treatment does not seem to be indicated in general. Subgroup-directed treatment has become a reality for acute myeloid leukemia in young children with Down's syndrome and in acute promyelocytic leukemia. In addition to tailoring therapy according to biologic features and especially monitoring treatment by measurements of minimal residual disease, targeted therapy for subgroups with activating mutations in receptor tyrosine kinases will further optimize the treatment of pediatric acute myeloid leukemia. Together with the development of many novel agents that have different mechanisms of action than the currently available anticancer agents, and improved supportive care, it is realistic that the prognosis of acute myeloid leukemia in children and adolescents will improve further in the next 5-10 years. PMID- 16221061 TI - Promising developments in prion immunotherapy. PMID- 16221064 TI - Glycoprotein D adjuvant herpes simplex virus vaccine. AB - Herpes simplex virus (HSV) Type-1 and -2 are common infections that can cause primary and recurrent herpes labialis and genitalis, as well as gingivostomatitis, keratoconjunctivitis, encephalitis, disseminated infections in immunocompromised persons and neonatal infections. Despite several decades of HSV vaccine development, no effective vaccine has been developed until recently. The following review of the genital HSV-2 glycoprotein D (gD2t, t is for truncated) subunit vaccine formulated with a new adjuvant (AS04) containing alum and 3-O deacylated monophosphoryl lipid A (MPL) provides a background in which to evaluate the vaccine as well as a brief review of other approaches to herpes vaccines. The gD2t-AS04 vaccine has been demonstrated to be safe in several large clinical trials. In two trials, the vaccine reduced genital herpes disease by 73 and 74%, but only in females with no previous HSV infection. A large ongoing trial in HSV seronegative females will provide additional data on protection from HSV disease and infection. PMID- 16221065 TI - Review of the Varilrix varicella vaccine. AB - Varicella zoster virus causes an acute infection that affects most children globally, but the age of infection can be greater in residents of tropical areas. It has generally been considered a mild disease, although there are accumulating data to show that it can cause significant morbidity and mortality in immunocompetent as well as immunocompromised children and adults. Oka-strain live attenuated varicella vaccines were developed in the 1970s. Varilrix developed by GlaxoSmithKline Biologicals (Rixensart, Belgium), is one of the vaccines produced and marketed in over 80 countries. Similar to the other Oka-strain vaccines, Varilrix is safe, immunogenic and efficacious in both immunocompromised and immunocompetent children and adults. PMID- 16221066 TI - Application of vaccine technology to prevention of Pseudomonas aeruginosa infections. AB - Development of an effective vaccine against the multiple presentations of Pseudomonas aeruginosa infection, including nosocomial pneumonia, bloodstream infections, chronic lung infections in cystic fibrosis patients and potentially sight-threatening keratitis in users of contact lenses, is a high priority. As with vaccine development for any pathogen, key information about the most effective immunologic effectors of immunity and target antigens needs to be established. For P. aeruginosa, although there is a role for cell-mediated immunity in animals following active vaccination, the bulk of the data indicate that opsonically-active antibodies provide the most effective mediators of acquired immunity. Major target antigens include the lipopolysaccharide O polysaccharides, cell-surface alginate, flagella, components of the Type III secretion apparatus and outer membrane proteins with a potentially additive effect achieved by including immune effectors to toxins and proteases. A variety of active vaccination approaches have the potential for efficacy such as vaccination with purified or recombinant antigens incorporating multiple epitopes, conjugate vaccines incorporating proteins and carbohydrate antigens, and live attenuated vaccines, including heterologous antigen delivery systems expressing immunogenic P. aeruginosa antigens. A diverse range of passive immunotherapeutic approaches are also candidates for effective immunity, with a variety of human monoclonal antibodies described over the years with good preclinical efficacy and some early Phase I and II studies in humans. Finding an effective active and/or passive vaccination strategy for P. aeruginosa infections could be realized in the next 5 to 10 years, but will require that advances are made in the understanding of antigen expression and immune effectors that work in different human tissues and clinical settings, and also require a means to validate that clinical outcomes achieved in Phase III trials represent meaningful advances in management and treatment of P. aeruginosa infections. PMID- 16221068 TI - Prevention of Staphylococcus aureus infections: advances in vaccine development. AB - Staphylococcus aureus is a ubiquitous bacterial species that causes serious disease in a minority of carriers, particularly in hospital settings. S. aureus disease is difficult to treat, and antibiotic-resistant strains have become common. Prevention of S. aureus disease would therefore be the best way to limit the morbidity and mortality caused by this organism, but its virulence is determined by a number of different factors, making design of a widely effective vaccine difficult. Here, various S. aureus virulence factors and attempts to develop vaccines or other protective drugs based on these factors are reviewed. In particular, the results of a Phase III clinical study of a vaccine directed at capsular polysaccharides types 5 and 8 are discussed. PMID- 16221067 TI - Vaccine prospects for amebiasis. AB - Entamoeba histolytica is a eukaryotic protozoan parasite and is the causative agent of amebic colitis and amebic liver abscess. Many insights into the innate and acquired immune responses to infection with E. histolytica have been made in recent years. These findings have provided a foundation for producing a vaccine that could help to prevent the initial establishment of infection in the intestinal wall. The galactose and N-acetyl-D-galactosamine-specific lectin on the surface of the ameba is an immunodominant molecule that is highly conserved and has an integral role in the stimulation of these immune responses. The structure of the lectin has been defined, and the heavy subunit with its cysteine rich region has been demonstrated in animal models to have some efficacy as a possible vaccine agent for prevention of amebic infection. Finding an ideal animal model of amebic intestinal infection has been difficult, but the C3H mouse and severe combined immunodeficient mouse-human intestinal xenograft models have both provided valuable insights into the first line of immune defense at the mucosal wall of the colon. Providing safe food and water to all people in the developing world is a formidable task that is not achievable in the foreseeable future. However, a vaccine for amebiasis could make a significant impact on the morbidity and mortality from the disease. Many components of the ameba are immunogenic and may serve as targets for a future vaccine, including the galactose and N-acetyl-D-galactosamine lectin, the serine-rich E. histolytica protein, cysteine proteinases, lipophosphoglycans, amebapores and the 29-kDa protein. PMID- 16221069 TI - Synthetic carbohydrate-based antitumor vaccines: challenges and opportunities. AB - The development of a clinically effective, carbohydrate-based antitumor vaccine is a longstanding ambition in the prevention and treatment of cancer. This review seeks to provide a discussion of some of the unique challenges facing this particular field of immunology. The authors present a historic account of their ongoing research program devoted to the development of fully synthetic, carbohydrate-based anticancer vaccines of clinical value. As will be seen, remarkable advances in carbohydrate and glycopeptide assembly techniques have allowed for the preparation of synthetic constructs of progressively increasing structural complexity. The authors describe the evolution of their synthetic carbohydrate program from first-generation constructs, which were monovalent in nature, to highly complex unimolecular multivalent vaccines, in which multiple carbohydrate antigens are displayed in the context of a single polypeptide backbone. It is the hope that each generation of vaccines represents a move closer to achieving the ultimate objective of developing broadly useful, robust anticancer vaccines. PMID- 16221070 TI - Development of cowpea mosaic virus-based vectors for the production of vaccines in plants. AB - Plant viruses are emerging as an attractive alternative to stable genetic transformation for the expression of foreign proteins in plants. The main advantages of using this strategy are that viral genomes are small and easy to manipulate, infection of plants with modified viruses is simpler and quicker than the regeneration of stably transformed plants and the sequence inserted into a virus vector will be highly amplified. One use of these virus expression systems is for vaccine production. Among plant viruses, cowpea mosaic virus makes an ideal candidate for the production of such vaccines because it grows extremely well in host plants, is very stable, and the purification of virus particles, if required, is straightforward. In this article, the authors review the progress made in the development of cowpea mosaic virus-based vectors for vaccine production, making use of two main approaches: epitope presentation and polypeptide expression. PMID- 16221071 TI - Dendritic cells: tools and targets for antitumor vaccination. AB - Dendritic cells are the most potent antigen-presenting cells of the immune system and represent a promising tool in therapeutic vaccination against cancer. Immunotherapy applying ex vivo-generated and tumor antigen-loaded dentritic cells has been successfully introduced in clinical vaccination protocols and has proven to be feasible and effective in some patients. A better understanding of how dentritic cells succeed to induce and modulate immunity is necessary to optimally exploit dentritic cells in anticancer vaccines. The authors will review novel insights in antigen loading, activation and migration of dentritic cells and their impact on the application of ex vivo-generated dentritic cell vaccines. In addition, novel means to exploit dentritic cells in cancer vaccines by loading and activation of dentritic cells directly in situ and possible obstacles that should be overcome to induce long-lasting immunity in therapeutic settings will be discussed. PMID- 16221072 TI - Memory cytolytic T-lymphocytes: induction, regulation and implications for vaccine design. AB - The design of vaccines that protect against intracellular infections or cancer remains a challenge. In many cases, immunity depends on the development of antigen-specific memory CD8+ T-cells that can express cytokines and kill antigen bearing cells when they encounter the pathogen or tumor. Here, the authors review current understanding of the signals and cells that lead to memory CD8+ T-cell differentiation, the relationship between the primary CD8+ T-cell response and the memory response and the regulation of memory CD8+ T-cell survival and function. The implications of this new knowledge for vaccine design are discussed, and recent progress in the development of lipidated peptide vaccines as a promising approach for vaccination against intracellular infections and cancer is reviewed. PMID- 16221073 TI - Vaccination of the immunocompromised child. AB - The development of vaccination is a major achievement in modern medicine. However, children treated with immunosuppression may not at all, or only in part, receive routine immunization due to uncertainty of its risks and effect. There is a substantial lack of pediatric studies concerning the efficacy and safety of vaccination in this patient group. Experience from similar adult groups and children with HIV infection can be used as a model for other disease categories. With increasing knowledge of the immunologic basis of vaccination and how immunosuppressive drugs interfere with the immune system, improved vaccines could be tailored, and adequate, individualized guidelines issued. PMID- 16221074 TI - Hunting for immune response regulatory genes: vaccination studies in infant twins. AB - The contribution of twin studies towards unraveling the complex mechanisms of multifactorial diseases is increasingly recognized. Recent twin studies using infant vaccination as a model for infectious diseases have confirmed the importance of host genetic factors as major regulators of the immune response. A combination of twin-based family studies and population-based association studies should lead to the identification of the specific genes involved. These genes and their products have the potential to be developed as targets for novel therapeutic and prophylactic agents against infectious diseases. PMID- 16221075 TI - Tick-borne encephalitis vaccines: past and present. AB - Vaccines to protect against tick-borne encephalitis (TBE) are produced by two manufacturers and are widely used in European and Asian countries, where TBE virus is endemic. General trends in vaccine development during recent decades and extensive postmarketing experience resulted in several modifications to their formulations and practical implications for use. Modifications were made to the production process, such as the change of the virus master bank from mouse brain to primary cells; to the excipients, especially the stabilizers and preservative; and to include formulations for children. Additionally, a rapid vaccination schedule has been developed for persons who require a fast onset of protection. Recent data from clinical studies and postmarketing surveillance indicate that both vaccines are safe, efficacious and interchangeable. Further (major) changes to formulation or alternative targets for vaccine development are not anticipated in the next 5 years. Recent serologic studies indicate that the persistence of protective immunity was longer than expected. Thus, recommendations for prolongation of TBE booster intervals have been made in several European countries, and a harmonization for booster recommendations is predicted within the European Union. Based on epidemiologic trends, the use of TBE vaccines will continue to increase in all age groups, including children. PMID- 16221076 TI - Are vaccination programs and isolate polymorphism linked to pertussis re emergence? AB - Whooping cough remains an endemic disease, and the re-emergence of pertussis in older children and adolescents has been reported in several countries, despite high vaccine coverage. Polymorphism of Bordetella pertussis has been observed over time, and some characteristics of pertussis isolates have gradually diverged from the vaccine strains. The present review summarizes the current knowledge on B. pertussis variability in countries with different vaccination programs and discusses its potential impact on the recently observed increased incidence of whooping cough. No direct association between B. pertussis isolate variability and vaccination programs has been observed to date, except for shifts from fimbriae Fim2 to Fim3. More likely explanations for the re-emergence of pertussis include the change in the epidemiology and transmission patterns of pertussis in highly vaccinated populations, and a shift of disease from young children to adolescents and adults due to waning protective immunity. PMID- 16221077 TI - Exploring human interaction and diet effects on the behavior of dogs in a public animal shelter. AB - This study examined the effects of 2 manipulations--a brief, regular period of human contact and diet--on the behavior of dogs confined in a public animal shelter. A behavioral battery designed to assess reactions to novel situations, and a test of responsiveness to an unfamiliar human were administered both prior to (pretest) and immediately following (posttest) the 8-week intervention period. Overall, the regular periods of increased human contact together with a diet that contained augmented levels of digestible protein, fat, calories, and animal derived ingredients reduced signs of behavioral reactivity from pretest to posttest. In some cases, the comparison diet appeared more effective, but only for dogs receiving minimal human interaction. The results indicate that a combination of human interaction and high quality diet may positively affect the behavior of dogs in animal shelters. PMID- 16221078 TI - Octopus senescence: the beginning of the end. AB - Senescence is a normal stage of an octopus's life cycle that often occurs before death. Some of the following symptoms typify it: lack of feeding, retraction of skin around the eyes, uncoordinated movement, increased undirected activity, and white unhealing lesions on the body. There is inter- and intraspecific variability. Senescence is not a disease or a result of disease, although diseases can also be a symptom of it. Both males and females go through a senescent stage before dying-the males after mating, the females while brooding eggs and after the eggs hatch. There are many aspects of octopus senescence that have not yet been studied. This study discusses the ecological implications of senescence. PMID- 16221079 TI - The effects of implementing a feral cat spay/neuter program in a Florida county animal control service. AB - In 1995, a county animal control service implemented a feral cat sterilization program with the goal of reducing the number of healthy cats euthanized, complaints, and the county's costs. The service collected data from a 6-year period both before and after the program's implementation. The service totaled the numbers of both cat and dog impoundments, surgeries, adoptions, euthanasias, and complaints for each year; standardized both sets of numbers on a per- 10,000 person basis to compare trends between dogs and cats; and calculated estimated costs for neutering versus impounding and euthanizing the feral cats. Changing from a policy of euthanasia of feral cats to support for trap-neuter-return did not result in an increase in the number of complaints or cat impoundments. The percentage of impounded cats euthanized decreased between the periods before and after the program began, and the percentage adopted increased. The ratio of complaints to impounds decreased between the 2 periods, and the ratio of surgeries to impounds increased. Impoundments of cats were relatively steady in spite of the continually increasing human population. Euthanasias decreased for both cats and dogs since 1997. Since 1996, complaints decreased for both. Surgeries for both cats and dogs increased over the 12 years. Adoptions for cats and dogs increased greatly in fiscal year 1998/1999. PMID- 16221080 TI - Long-term outcome after treatment of feline inappropriate elimination. AB - Urination and defecation outside the litter box is one of the most common behavior problems reported by cat caretakers and the most common behavioral reason that results in the relinquishment of cats to shelters. A physical examination, laboratory tests, and a thorough behavioral history are necessary to differentiate between the diagnoses of disease, marking, and inappropriate elimination. This study followed up on the outcome of cats who were treated for inappropriate elimination. Fifty-eight caregivers completed a telephone questionnaire 12 to 54 months after treatment was initiated. Twenty-seven (47%) of caregivers felt their cat was cured, 15 (26%) felt their cat was much better, 8 (14%) slightly better, 6 (10%) the same, and 2 (3%) worse. Forty caregivers (67%) observed a 90% to 100% reduction in frequency. There was a significant association between the caregivers' perception of treatment outcome and percentage reduction in frequency. There was a significant association between caregivers compliance and outcome. The results of this study give caregivers a valid and promising alternative to relinquishment for inappropriate elimination problems. PMID- 16221081 TI - The effects of caretaker-primate relationships on primates in the laboratory. AB - As contact with caretakers is likely to make up the majority of human-primate interactions in laboratories, caretakers represent an important influence in the lives of captive primates. The aim of this study was to determine how caretaker primate relationships affected the behavior of primates in the laboratory. We examined whether stump-tailed macaques (Macaca arctoides) who were evaluated by caretakers as being either friendly or unfriendly differed in the quality and quantity of interactions with their caretakers during husbandry procedures and in their behavior at times of high and low levels of caretaker activity. Results revealed that animals who had friendly relationships with caretakers were less disturbed by routine husbandry procedures, approached caretakers more often, and were willing to accept food offered by caretakers compared with animals considered unfriendly toward their caretakers. The study concluded that the quality of the primate-caretaker relationship may have an important impact on behavior and may have implications for the well-being of animals and caretakers, as both can benefit from positive feedback from one another. PMID- 16221082 TI - The myth of the aggressive monkey. AB - Captive rhesus macaques are not naturally aggressive, but poor husbandry and handling practices can trigger their aggression toward conspecifics and toward the human handler. The myth of the aggressive monkey probably is based on often not taking into account basic ethological principles when managing rhesus macaques in the research laboratory setting. PMID- 16221088 TI - Renal tubular function in children with beta-thalassemia minor. AB - BACKGROUND: beta-thalassemia minor is a common heterozygous haemoglobinopathy that is characterized by both microcytosis and hypochromia. It requires no treatment. It has been postulated that low-grade haemolysis, tubular iron deposition and toxins derived from erythrocytes might cause renal tubular damage in adult patients with beta-thalassemia minor. Our aim was to investigate the renal tubular functions in children with beta-thalassemia minor and to determine its possible harmful effects. METHODS: The study was conducted on 32 children (14 female and 18 male) at the age of 5.8 +/- 3.1 years (range 2-14 years) with beta thalassemia minor. The patients were classified as anaemic (haemoglobin (Hb) 11 g/dL) (Group 2, n = 18). A control group was formed with 18 healthy children whose ages and sexes match those in other groups (Group 3, n = 18). Fractional excretion of sodium (FE(Na), %), fractional excretion of magnesium (FE(Mg), %), fractional excretion of uric acid (FE(UA), %) and tubular phosphorus reabsorption (TPR,%) were calculated with standard formulas. Urinary calcium excretion (mg/kg per 24 h), zinc (Zn) (microg/dL), glucosuria (mg/dL), beta-2 microglobulin (mg/dL) and N-acetyl-beta-D glycosaminidase (NAG, U/mmol creatinine) levels were measured through biochemical methods. RESULTS: There was no statistically significant difference among the three groups in terms of the results of FE(Na) (%), FE(Mg) (%), FE(UA) (%), TPR (%), calciuria (mg/kg per 24 h), NAG, urine Zn, proteinuria, glucosuria or urine beta- 2 microglobulin levels (P > 0.05). CONCLUSION: On the contrary of children with beta-thalassemia major, renal tubular dysfunction has not been determined in children with beta-thalassemia minor in the present study. PMID- 16221089 TI - Paraquat-induced Fanconi syndrome. AB - The ingestion of paraquat, a non-selective herbicide, can be fatal in humans. Paraquat is toxic to multiple organs, including the kidney, heart, gastrointestinal tract and central nervous system. Although paraquat has been established as one cause of acute tubular necrosis, Fanconi syndrome presenting as severe hypophosphataemia after paraquat intoxication has not been reported. Here, we report the case of a 44-year-old Korean woman who presented with generalized proximal tubular dysfunction including aminoaciduria, phosphaturia and glycosuria after paraquat intoxication. We found that severe hypophosphataemia induces deep drowsiness. Renal biopsy findings indicated the presence of acute tubular necrosis that may be reversible. PMID- 16221091 TI - Haemodialysis-unresponsive blood pressure: cardiovascular mortality predictor? AB - AIM: The importance of 'conventional' cardiovascular risk factors in haemodialysis (HD) patients has been questioned following evidence that pre-HD blood pressure (BP) might be inversely related to mortality in contrast to post HD BP. To evaluate this reverse BP epidemiology in HD patients, HD-induced BP changes were compared with aortic pulse wave velocity (PWV), an independent predictor of cardiovascular mortality. METHOD: Aortic PWV was evaluated in a limited care HD cohort, all of whom were asymptomatic of cardiovascular disease. RESULTS: Of 47 limited care patients, 45% were classified as HD responsive, with a 17% fall in mean arterial pressure compared with a 6% increase in the HD unresponsive group. While there were no significant differences between the two groups in traditional vascular disease risk factors or interdialytic weight loss, PWV was significantly elevated in the HD-unresponsive group (12.9 +/- 2.7 compared with 10.8 +/- 2.9; P < 0.05). Furthermore, there was a positive correlation between the change in BP during HD and PWV in all subjects (r = 0.500; P < 0.001 for systolic blood pressure (SBP), a correlation that also existed for diastolic blood pressure (DBP) (P < 0.01). CONCLUSION: This study suggests that patients with HD-unresponsive BP are more likely to have vascular disease and this association between PWV and HD-induced BP changes might partly explain the apparent paradox of pre- and post-HD BP with mortality. It is proposed that a population with elevated post-HD BP is more likely to be composed of subjects with vascular disease (overt or occult), in contrast to a group with high pre-HD BP, which will be more heterogeneous. PMID- 16221092 TI - Course of C-reactive protein during continuous peritoneal dialysis-associated peritonitis. AB - OBJECTIVE: C-reactive protein (CRP) levels increase during peritonitis. Little is known about the extent and duration of CRP elevation. PATIENTS AND METHODS: Patients on continuous peritoneal dialysis (CPD) therapy had a baseline CRP value at the start of the study. CRP values were determined at 48 h, 2 weeks and 4 weeks after an episode of peritonitis developed in the 13 months after the study started. RESULTS: The demographic values and comorbid diseases of the patients developing and not developing peritonitis were not significantly different. Patients who developed peritonitis had a mean +/- SD CRP value of 15.0 +/- 11.1 mg/L compared with 15.7 +/- 9.9 mg/L in patients without peritonitis. The CRP value rose to 118.35 +/- 96.86 mg/L at 48 h, and decreased to 34.5 +/- 43.6 mg/L at 2 weeks and to 30.1 +/- 39.7 mg/L at 4 weeks after the onset of peritonitis. All types of peritonitis were associated with a significant increase in CRP at 48 h. Nine patients had CRP elevations above baseline at 4 weeks; seven patients had active medical issues. CONCLUSION: CRP values become significantly elevated during peritonitis. Most patients have a decrease in the CRP values toward baseline values at 4 weeks following the onset of peritonitis while 20% of patients had continued elevations. The significance of the elevations is unclear. Given the associations of CRP with cardiovascular mortality in both uremic and non-uremic patients and the association of peritonitis with mortality, the role of this inflammatory marker deserves further evaluation. PMID- 16221090 TI - Relationship between iron replacement and hepatic functions in hepatitis C virus positive chronic haemodialysis patients. AB - AIM: To investigate the effects of intravenous (i.v.) iron replacement on hepatic functions of hepatitis C virus (HCV)-positive haemodialysis patients. METHODS: The present retrospective study included 89 HCV-positive and 57 HCV-negative haemodialysis patients. Alanine aminotransferase (ALT) levels were accepted as sustained high if the last three values were >/=20 U/L. All patients and the HCV positive group were dichotomised into subgroups by the median for dialysis duration, the amounts of i.v. iron administered per year and totally. RESULTS: Sustained high levels of ALT were significantly more frequent in the HCV-positive group (P < 0.001). In HCV-positive patients, the subgroup with ALT levels >/=20 U/L had significantly higher serum iron levels and mean amounts of i.v. iron administered per year and totally (P < 0.001) and the subgroup with the high mean total amount of i.v. iron had significantly higher serum ALT and iron levels (P < 0.001). Significant positive correlations were found in HCV-positive patients between ALT and serum iron levels (P < 0.001), as well as between ALT both with the mean amounts of i.v. iron administered per year (P = 0006) and totally (P = 0.015). Regression analysis showed that the main parameters effecting ALT were the serum iron level (P < 0.0001) and the mean amount of parenteral iron administered per year (P = 0.032). CONCLUSION: We conclude that parenteral iron replacement might contribute to hepatocellular injury in HCV-positive haemodialysis patients. PMID- 16221093 TI - WHO class-specific equations using height for predicting body weight: crude indicator for dry weight in haemodialysis patients. AB - AIM: Body weight (BW) might be related to total body water, and the difference between a patient's actual BW and ideal BW (IBW) might be the volume marker. However, there has been no information about the association between IBW and dry weight (DW) in haemodialysis (HD) patients. METHODS: First, we analysed the relationship between DW and IBW in 51 HD patients. The IBW was calculated by 21 x Height (Ht)(2). Weight status was analysed by the WHO classification. Second, in 12 436 controls, linear equations using Ht(2) were sought to predict the BW in each sex and WHO class. Third, using these equations, predicted BW (PW) was compared with DW in each WHO class at the initiation and after 1 year in 619 new HD patients, retrospectively. RESULTS: Among 51 HD patients, 38 were normal weight in whom there was no difference between DW and IBW. In each sex and WHO class of the 12 436 controls, linear equations using Ht(2) were developed to predict BW. These equations were applied to 619 new HD patients. In males, there were no differences between PW and DW in underweight (UW), overweight (OW), obese (OB) and extremely obese (EOB) patients at the initiation of the HD. In females, there were no differences between PW and DW in OW patients. Despite no statistical differences, there were wide ranges of distribution from -6 to 6 kg between PW and DW. CONCLUSIONS: BW had a linear relationship with Ht(2) and might be predictable by the WHO class-specific equation using Ht(2). These equations might be useful as a crude indicator of DW in HD patients. PMID- 16221094 TI - Cardiac troponin I and beta 2 microglobulin as risk factors for early-onset atherosclerosis in patients on haemodialysis. AB - AIM: To investigate the associations of different risk factors with carotid artery intima-media thickness (C-IMT) in non-diabetic haemodialysis (HD) patients who had no clinical evidence of atherosclerosis. METHODS: Seventy-two HD patients (43 men, 29 women; mean age: 34.5 +/- 10.6 years; mean time on HD: 47.9 +/- 40.0 months) and 40 age- and sex-matched healthy controls (26 men, 14 women; mean age: 35.5 +/- 7.1 years) participated in the study. The relationship between C-IMT and haematocrit-corrected erythrocyte sedimentation rate (Hct-corrected ESR), beta 2 microglobulin (beta2M) and serum cardiac troponin I (cTnI) levels beyond C reactive protein (CRP), lipid profile and lipoprotein(a), fibrinogen, homocysteine and left ventricular hypertrophy (LVH) were examined. RESULTS: Mean C-IMT of the HD patients was significantly greater than that of the control subjects (0.59 +/- 0.06 vs 0.53 +/- 0.07 mm, P = 0.002). C-IMT of patients was positively correlated with age (r = 0.33), body mass index (r = 0.40), Hct corrected ESR (r = 0.37), CRP (r = 0.34), beta2M (r = 0.34), cTnI (r = 0.26), triglyceride (r = 0.26) and fibrinogen (r = 0.28) levels (P < 0.05 for all). The mean C-IMT was significantly greater in patients with LVH than it was in those without LVH (P = 0.004). In multivariate regression analysis, age (P = 0.02), beta2M (P = 0.001), log-transformed CRP (P = 0.03) and LVH (P = 0.01) were independently related with C-IMT. CONCLUSION: Besides well-known cardiovascular (CV) risk factors, cTnI and beta2M were related with C-IMT in that they may have important roles in early-onset atherosclerosis in this high-risk population. PMID- 16221095 TI - Vitamin A and zinc status in patients on maintenance haemodialysis. AB - BACKGROUND: The objective of this study was to assess the vitamin A and zinc serum levels in patients undergoing haemodialysis (HD) in the city of Recife, in the north-eastern region of Brazil. METHODS: The study comprised 55 patients and 28 healthy controls. The retinol and zinc serum concentrations were analysed by using high-performance liquid chromatography (HPLC) and atomic absorption spectrophotometry, respectively. RESULTS: The mean retinol serum concentration in patients (2.50 +/- 0.86 micromol/L) was significantly greater (P < 0.001) than that found in controls (1.26 +/- 0.86 micromol/L). The retinol serum levels in the patients were as follows: 47.3% of the patients had elevated levels (>/= 2.24 and < 3.50 micromol/L); 16.4% of the patients had serum levels >/= 3.50 micromol/L, which indicated hypervitaminosis; and 9.1% of the patients had serum levels below the normal range (<1.05 micromol/L), a rate that among the controls was 42.9% (P < 0.01). In regard to zinc, the serum levels found in the patients (10.59 +/- 3.12 micromol/L) were similar to those found in the controls (11.43 +/ 2.82 micromol/L) (P > 0.05). Although 49.1% of the patients and 35.7% of the controls were classified as deficient in zinc, this difference was not statistically significant (P > 0.05). RESULTS: The results identified a high prevalence of zinc deficiency in the groups studied, and point to a trend towards more elevated retinol serum levels in patients undergoing dialysis as compared with those in healthy controls. PMID- 16221096 TI - Apoptosis of circulating lymphocyte in rats with unilateral ureteral obstruction: role of angiotensin II. AB - BACKGROUND: Unilateral ureteral obstruction (UUO) could induce increased renal angiotensin II (ANG II), which enhances apoptosis of renal tubular cells and renal tissue loss. Systemic ANG II is also increased in UUO. There are no data available about whether UUO can induce apoptosis of circulating lymphocytes or not. METHODS: UUO or sham-operated male Wistar rats (n = 8 in each group) were fed a drinking solution containing water, angiotensin II receptor type 1 antagonist (ARA; losartan, 500 mg/L) or angiotensin-converting enzyme inhibitor (ACEI; enalapril: 200 mg/L) for 1 day or 7 days. Blood samples were collected and circulating lymphocyte cells were separated. The apoptotic cells were detected by in situ terminal deoxynucleotidyl transferase (TdT assay)-mediated digoxigenin/antidigoxigenin conjugated fluorescein method and counted under a fluorescence microscope. The apoptotic index was calculated. RESULTS: UUO caused marked increases in the apoptotic index of circulating lymphocytes in UUO rats at both 1 day and 7 days when compared with the respective sham groups (P < 0.001). Neither ARA nor ACEI treatment had an effect on the apoptotic index values in the UUO rats at 1 day. In the UUO rats at 7 days, the apoptosis of circulating lymphocytes was markedly decreased from 29.2 +/- 2.7% to 11.9 +/- 2.7% (P < 0.01) in the ARA-treated rats and to 7.6 +/- 2.7% (P < 0.001) in the ACEI-treated rats. CONCLUSION: UUO, via stimulation of ANG II, could promptly enhance apoptosis of circulating lymphocytes. The apoptosis persisted throughout the 7 days of the study. Prolonged UUO would impair lymphocyte cell immunity and the host defense mechanism. Continuous treatment with either ARA or ACEI could abrogate ANG II stimulated circulating lymphocyte apoptosis. PMID- 16221097 TI - Partial depletion of macrophages by ED7 reduces renal injury in Adriamycin nephropathy. AB - BACKGROUND: Because macrophages are considered to be possible effectors of disease in Adriamycin (ADR) nephrosis, we hypothesized that depletion of macrophages might protect against the initiation of renal injury. In the present study, a monoclonal antibody (ED7) directed against CD11b/CD18 integrin, which is expressed by macrophages, was used to investigate the pathogenetic effects of macrophages in ADR nephropathy. METHODS: Male Wistar rats were treated with ED7 antibody, starting 1 day prior to ADR (7.5 mg/kg) treatment, or 7 days post-ADR when overt proteinuria was established. RESULTS: Circulating ED7-positive cells were reduced by approximately 30% in rats with ADR nephrosis by the ED7 antibody, while the number of macrophages in the renal cortex of ADR rats was reduced by nearly 50% with the ED7 treatment, whether administered before or after ADR. Creatinine clearance was significantly ameliorated by ED7 when commenced pre-ADR (P < 0.05), but not when commenced post-ADR (P = NS) in comparison to untreated ADR rats. However, proteinuria was not alleviated by either ED7 treatment. Morphometric analysis showed less glomerular sclerosis when ED7 was commenced pre ADR compared with ADR alone (P < 0.01), but not when commenced post-ADR (P = NS). Tubular atrophy was reduced by ED7 when it was commenced pre-ADR (tubular cell height and tubular diameter: P < 0.01 and P < 0.001, respectively), as was interstitial expansion (P < 0.01) compared with ADR alone. Cortical macrophage infiltration was reduced by 50% compared with ADR alone by the ED7 commenced before or after ADR. The number of cortical CD4+ T cells fell with ED7 starting pre-ADR, but not with the ED7 treatment commencing after ADR. CONCLUSION: Partial macrophage depletion starting before but not after ADR protected both renal function and structure in this model of chronic proteinuric renal disease. PMID- 16221098 TI - TGF-beta and TNF-alpha producing effects of losartan and amlodipine on human mononuclear cell culture. AB - AIM: The modulation of cytokine release, which affects adhesion of leucocytes to endothelial cells, and proliferation of peripheral blood mononuclear cells with antihypertensive drugs was explored. METHOD: In the present study, mononuclear cells were incubated with losartan and amlodipine at concentrations of 10(-6), 10(-5) and 10(-4) mol/L for 6 h. Transforming growth factor (TGF)-beta and tumour necrosis factor (TNF)-alpha levels were measured. Proliferation of mononuclear cells were assessed at the same concentrations of amlodipine and losartan with the methylthiazoletetrazolium (MTT) test. RESULTS: Amlodipine was found to induce TGF-beta synthesis from mononuclear cells with increasing concentrations, while it was found to inhibit TNF-alpha secretion with increasing concentrations. In contrast, losartan was found to induce TGF-beta and TNF-alpha secretion with increasing concentrations. CONCLUSION: Anti-atherosclerotic effects of amlodipine and losartan might be through increased secretion of TGF-beta from mononuclear cells. Different results at different concentrations might be due to the pharmocokinetic differences of these drugs. PMID- 16221099 TI - Renal plasma flow, filtration fraction and microalbuminuria in hypertensive patients: effects of chronic smoking. AB - INTRODUCTION: Albumin excretion rate is usually increased in people who smoke, but the physiological basis of this phenomenon is not fully understood. METHODS: The effect of chronic smoking on renal haemodynamics was studied in a cohort of 66 men. Twenty-seven were smokers and 36 were hypertensive. In all subjects, the albumin excretion rate was evaluated; in hypertensive patients, a renoscintigraphic evaluation of renal plasma flow and glomerular filtration were carried out and the filtration fraction was calculated. RESULTS: The hypertensive smoking population presented an increased urinary albumin excretion rate in comparison with hypertensive non-smoking patients. No significant differences were found for the mean values of renal plasma flow, the glomerular filtration rate and the filtration fraction between hypertensive smokers and hypertensive non-smoking patients. CONCLUSIONS: In hypertensive patients, smoking does not modify typical renal haemodynamic changes of arterial hypertension; however, it significantly increases the albumin excretion rate. PMID- 16221100 TI - Plasma and urinary adrenomedullin levels in children with acute pyelonephritis. AB - AIM: Adrenomedullin (AM), a novel peptide recently isolated from pheochromocytoma, eliciting vasorelaxing activity, is the strongest among all known peptides. AM has been detected in the adrenal medulla, cardiac tissue, lung and kidney. Immunohistochemical studies have demonstrated the localization of AM in glomeruli, tubules and collecting cells of the kidney. Clinically, plasma and urinary AM levels are altered in patients with different renal disease. The present study aims to determine plasma and urinary AM levels in children with acute pyelonephritis (APN) and compare the results with a control group. MATERIALS AND METHODS: The study group was comprised of 19 patients with APN aged 11.6 +/- 3.7 months (range, 6-18 months) and the control group consisted of 16 cases aged 11.5 +/- 3.2 months (range, 7-16 months). Acute pyelonephritis was diagnosed by clinical, laboratory and imaging methods. Plasma and urinary AM levels were measured by high performance liquid chromotography (HPLC). RESULTS: The plasma AM levels were lower in APN patients (33.40 +/- 2.27 pmol/mL) than in the control group (43.76 +/- 4.27 pmol/mL) (P < 0.001), whereas the urinary AM levels were higher in APN patients (248.58 +/- 140.63 pmol/mg urinary creatinine) than in the control group (49.42 +/- 45.23 pmol/mg) (P < 0.001). Coefficients of correlation between urinary AM levels and C-reactive protein and white blood cells were statistically significant (r = 0.472, P = 0.041; r = 0.555, P = 0.014, respectively). CONCLUSION: Adrenomedullin, a smooth muscle relaxant peptide that is synthesized in urinary tract tissue might have a role in acute pyelonephritis. However, the importance of AM in the pathogenesis of acute pyelonephritis remains to be determined by further detailed studies. PMID- 16221101 TI - Antineutrophil cytoplasmic autoantibodies in patients with systemic lupus erythematosus recognize a novel 69 kDa target antigen of neutrophil granules. AB - OBJECTIVE: Antineutrophil cytoplasmic autoantibodies (ANCA) were found in patients with systemic lupus erythematosus (SLE). Cathepsin G and lactoferrin were the major target antigens. However, some ANCA-positive sera did not recognize either of them. The present study was to investigate the unknown target antigens of ANCA in patients with SLE and their clinical significance. METHODS: Sera were collected from 72 patients with SLE. ANCA were detected in both indirect immunofluorescence and antigen-specific enzyme-linked immunosorbent assay (ELISA). Mixed neutrophil granules were separated from normal human peripheral neutrophils; soluble acid extracts in non-reducing conditions were used as antigens in western blot analysis to detect ANCA. RESULTS: SLE sera could blot a few bands. Interestingly, 14/72 (19.4%) sera recognized a novel 69 kDa protein band and 10/72 (13.9%) sera recognized the 55 kDa protein band, which might be bactericidal/permeability-increasing protein (BPI). The 69 kDa target antigen was different from the known target ANCA antigens such as cathepsin G and lactoferrin. Further study revealed that the percentages of patients with photosensitivity and oral ulcer in the anti-69 kDa autoantibodies-positive group were significantly higher than those in the anti-69 kDa autoantibodies-negative group (57.1%vs 10.3%, P < 0.005 and 50.0%vs 12.1%, P < 0.05, respectively). CONCLUSIONS: A 69 kDa protein in human neutrophil granules was identified as a novel target antigen of ANCA in patients with SLE. The anti-69 kDa autoantibodies might be associated with photosensitivity and oral ulcer in patients with SLE. PMID- 16221102 TI - Presentation, prognosis and outcome of IgA nephropathy in Indian adults. AB - BACKGROUND: IgA nephropathy (IgAN) is not well characterized in India. This retrospective study of 478 patients with IgAN was performed to clarify the presenting features, prognostic factors and the renal survival rates of the disease. METHODS: Three hundred and forty-seven patients who had been followed on average for 27 months after diagnosis were divided into two groups based on renal function at diagnosis. In group 1 (229 patients), the creatinine clearance estimated by the Modification of Diet in Renal Disease formula was <85 mL/min and in group 2 (118 patients) it was >/=85 mL/min. RESULTS: The predominant modes of presentation were nephrotic syndrome, hypertension and renal failure. Twenty-nine percent of patients had more than a 20% decline in renal function at the last follow up. Multivariate analyses with stepwise logistic regression identified hypertension (odds ratio (OR) 3.5), nephrotic range proteinuria (OR 3.4) and sclerosed glomeruli on biopsy (OR 4.1) to be independently associated with progression in group 1 and hypertension (OR 2.3) in group 2. Seventeen percent of patients progressed to end-stage renal disease (ESRD). Using multivariate analysis by the Cox model, four risk factors for developing ESRD were identified: hypertension (hazard ratio (HR) 3.1); nephrotic proteinuria (HR 1.9); interstitial fibrosis (HR 2.5); and sclerosed glomeruli (HR 1.8). The renal survival rates at 1, 5 and 10 years were 84, 55 and 33%, respectively, with a median renal survival of 61 months from the time of biopsy. CONCLUSION: The relatively rapid rate of progression of IgAN in India is suggestive towards a 'malignant' nature of the disease in this country. PMID- 16221104 TI - Parathyroidectomy in the calcimimetic era. AB - With the introduction of the calcimimetic cinacalcet HCl, some patients who would previously have undergone parathyroidectomy are likely to remain on medical therapy. Data is available on complication rates and some important outcome measures of parathyroidectomy, but the efficacy of calcimimetics to influence patient-based endpoints such as cardiovascular mortality and renal osteodystrophy has not been established. Nevertheless, cinacalcet HCl has been demonstrated to improve levels of calcium, phosphate, the calcium phosphate product and parathyroid hormone (PTH). Based on available data, parathyroidectomy is proposed as the preferred treatment option when averaged levels of intact PTH (iPTH) exceed 85-95 pmol/L despite optimal therapy. When iPTH levels exceed 50 pmol/L, parathyroidectomy should be considered if levels of serum calcium, phosphate or the calcium phosphate product are above established target ranges or when patients with established osteoporosis have progressive loss of bone mineral density. Because the currently-recommended biochemical targets are difficult to achieve and maintain for many patients on dialysis, parathyroidectomy rates are likely to increase if these management proposals are followed. This highlights the need for prospective studies with 'hard' endpoints, to establish evidence based roles for parathyroidectomy and calcimimetic therapy. PMID- 16221103 TI - Randomized controlled trial of pulse intravenous cyclophosphamide versus mycophenolate mofetil in the induction therapy of proliferative lupus nephritis. AB - BACKGROUND: The aim of the present study was to evaluate the efficacy of mycophenolate mofetil in the induction therapy of proliferative lupus nephritis. METHODS: Forty-four patients from eight centres with newly diagnosed lupus nephritis World Health Organization class III or IV were randomly assigned to either mycophenolate mofetil (MMF) 2 g/day for 6 months or intravenous cyclophosphamide (IVC) 0.75-1 g/m(2) monthly for 6 months in addition to corticosteroids. RESULTS: Remission occurred in 13 out of 25 patients (52%) in the IVC group and 11 out of 19 patients (58%) in the MMF group (P = 0.70). There were 12% in the IVC group and 26% in the MMF group that achieved complete remission (P = 0.22). Improvements in haemoglobin, the erythrocyte sedimentation rate, serum albumin, serum complement, proteinuria, urinary activity, renal function and the Systemic Lupus Erythematosus Disease Activity Index score were similar in both groups. Twenty-four follow-up renal biopsies at the end of therapy showed a significant reduction in the activity score in both groups. The chronicity index increased in both groups but was only significant in the IVC group. Adverse events were similar. Major infections occurred in three patients in each group. There was no difference in gastrointestinal side-effects. CONCLUSIONS: MMF in combination with corticosteroids is an effective induction therapy for moderately severe proliferative lupus nephritis. PMID- 16221105 TI - Effects of late referral to a nephrologist in patients with chronic renal failure. AB - BACKGROUND: We lack information about the role of late diagnosis of end-stage renal disease (ESRD), late nephrological referral and its impact on biochemical variables and first hospitalization in East Anatolia, Turkey. METHODS AND RESULTS: For a total of 101 ESRD patients, dialysis was initiated between January 1998 and December 2002 at the Yuzuncu Yil University Hospital. Early referral (ER) and late referral (LR) were defined as the time of first referral or admission to a nephrologist greater or less than 12 weeks, respectively, before initiation of haemodialysis (HD). RESULTS: The need for urgent dialysis was less among the early referral cases compared with the late referral cases (P = 0.03). Patients with LR started dialysis with lower levels of haemoglobin (8.6 vs 9.5 g/dL, P < 0.05) bicarbonate (16 vs 12 mEq/lt, P < 0.03) and albumin (2.9 vs 3.29 mg/dL, P < 0.02) and with higher serum levels of blood urea nitrogen (173 vs 95 mg/dL, P < 0.001), creatinine (10 vs 7.9 mg/dL, P < 0.001) and potassium (5.3 vs 4.8, P < 0.04). Hospitalization duration beginning at dialysis was significantly longer in the LR group (27.3 +/- 24) compared with the ER group (13.4 +/- 7.5, P < 0.001). When the groups were compared in terms of distance between the patients home and hospital, there were significantly more patients living far away from hospital (i.e. >100 km) in the LR group compared with the ER (P < 0.0001) group. CONCLUSION: Early referral to a nephrology unit and/or early diagnosis of ESRD results in better biochemical variables, shorter first hospitalization length and a higher percentage of elective construction of AVF and the availability to start with an alternative dialysis modality (i.e. CAPD). PMID- 16221107 TI - 'Flexible' or 'lifestyle' dialysis: is this the way forward? AB - BACKGROUND: Despite the advent of two new dialysis options, nocturnal home haemodialysis and short daily haemodialysis, many units are yet to build them into the modalities on offer to end-stage renal failure patients. The reasons behind this inertia are complex but primarily include anxieties about workload, budgetary implications and outcome data. METHOD: The Geelong dialysis programme, where both nocturnal home haemodialysis and short daily haemodialysis are offered, is compared with Australian and New Zealand national profiles. RESULTS: Significant profile differences emerge when comparing sessions/week and h/week between the three groups. Most Australian (92.93%) and New Zealand (95.07%) haemodialysis patients dialyse for three sessions/week. This contrasts to Geelong where only 73.6% dialyse for three sessions/week. 18.8% of Geelong haemodialysis patients versus 1.8% (Australia) and 0.9% (New Zealand) dialyse for five or more sessions/week. Australia and New Zealand follow similar h/session patterns although more Australians (44.2%) dialyse for 4 h and fewer (24.2%) for 5 h than their New Zealand counterparts (39.6% and 29.8%, respectively), and few dialyse outside the 3.5-5 h window. In contrast, 6.7% of Geelong patients dialyse for 2 2.5 h/session versus Australia (0.9%) and New Zealand (0.2%). This represents the Geelong short daily dialysis programme. More Geelong patients (>15%) dialyse >/=8 h/week and represent the Geelong nocturnal home haemodialysis programme. CONCLUSION: The flexible Geelong programme has been supported without exceeding the budget applied to a conventional dialysis programme with the same patient numbers. PMID- 16221106 TI - Losartan reduces the costs of diabetic end-stage renal disease: an Asian perspective. AB - OBJECTIVE: To evaluate losartan and conventional antihypertensive therapy (CT) compared with CT alone on the cost associated with end-stage renal disease (ESRD) in Hong Kong, Japan, Korea, Malaysia, Singapore and Taiwan. METHODS: Reduction of end-points in non-insulin-dependent diabetes mellitus with the angiotensin II antagonist losartan (RENAAL) was a multinational, double-blind, randomized, placebo-controlled trial to evaluate the renal protective effects of losartan on a background of CT in patients with type 2 diabetes and nephropathy. The primary composite end-point was a doubling of serum creatinine, ESRD or death. Data on the duration of ESRD for the Asian subgroup of patients enrolled in RENAAL were used to estimate the economic benefits of slowing the progression of nephropathy. The cost associated with ESRD was estimated by combining the number of days each patient experienced ESRD with the average daily cost of dialysis from the third party payer perspective in Hong Kong, Japan, Korea, Malaysia, Singapore and Taiwan. Total cost, converted to US dollars, was the sum of ESRD and losartan costs. RESULTS: Losartan plus CT reduced the number of days with ESRD by 37.9 per patient over 3.5 years compared with CT alone. This reduction in ESRD days resulted in a decrease in the cost associated with ESRD, which ranges from $910 to $4346 per patient over 3.5 years across the six countries or regions. After accounting for the cost of losartan, the reduction in ESRD days resulted in net savings in each of the six countries or regions, ranging from $55 to $515 per patient. CONCLUSION: Treatment with losartan in patients with type 2 diabetic nephropathy not only reduced the incidence of ESRD among Asian patients, but resulted in direct medical cost savings in countries or regions representing Asia. PMID- 16221108 TI - Renal involvement in bone marrow transplantation. AB - Bone marrow transplantation (BMT) is an effective therapeutic strategy for leukaemic malignancies and depressed bone marrow following cancer. However, its side effects on kidneys have been reported. Some drugs and irradiation are also suggested to be nephrotoxic. It is well known that haemolytic uraemic syndrome (HUS) after BMT develops as late-onset BMT nephropathy. Cyclosporine A (CsA) is a possible cause. Radiation nephropathy shows changes that are similar to the histology of HUS. These findings suggest that endothelial damage is closely associated with the pathogenesis of post-BMT nephropathy. Recently, some patients have developed glomerulonephritis accompanied by graft-versus-host disease (GVHD) after BMT. In these patients immune deposits are found mainly in subepithelium and mesangium equal to those of secondary membranous glomerulonephritis. A murine experimental model of GVHD manifests similar symptoms and histological changes to those of actual patients and may suggest the pathogenesis of glomerulonephritis. PMID- 16221109 TI - Apparent Addison's disease following ileostomy. PMID- 16221110 TI - Renal failure from wasp stings: an appraisal on the previous reported Thai cases. PMID- 16221113 TI - The CARI guidelines. Biochemical and haematological targets guidelines. Haemoglobin. PMID- 16221114 TI - The CARI guidelines. Deceased kidney donor suitability guidelines. PMID- 16221118 TI - The CARI guidelines. Evaluation of renal function. PMID- 16221124 TI - The CARI guidelines. Acceptance into dialysis guidelines. PMID- 16221128 TI - The CARI guidelines. Dialysis adequacy (HD) guidelines. PMID- 16221134 TI - The CARI guidelines. Dialysis adequacy (PD) guidelines. PMID- 16221138 TI - Age and skin structure and function, a quantitative approach (I): blood flow, pH, thickness, and ultrasound echogenicity. AB - BACKGROUND/PURPOSE: The aging process has been studied with fervor recently, given our shifting demographics. Since age's effects are so manifest in skin's appearance, structure, mechanics, and barrier function, it is not surprising that much effort has been placed in research to better understand them. Quantitative measurements permitted by bioengineering have allowed us to objectively and precisely study aging skin. These overviews piece together the immense amounts of information that have emerged from recent technological advances in dermatological research in order to develop a unified understanding of the quantitative effects of age on the skin. METHODS: We performed a literature on age-related changes in blood flow, pH, skin thickness, and ultrasound imaging data, searching Pub-med, Em-Base, Science Citation Index, and the UCSF dermatological library's collection of books on the topic of aging skin. RESULTS: Despite the many tools and techniques available for quantitative analysis of skin, age studies are often conflicting, especially in the areas of blood flow and skin thickness. Trends indicate that blood flow may decrease with age, especially in sites exposed to the environment. pH apparently varies little until the age of 70, after which it declines. Skin thickness data are difficult to interpret; while the stratum corneum is generally accepted to maintain its thickness during aging, dermal, epidermal, and whole skin thickness changes are controversial. Ultrasound reveals the appearance of a subepidermal low echogenic band that thickens with age, especially in environmentally exposed areas. Some studies also indicate the presence of an echogenic band in the lower dermis which thins with increased age. However, the whole dermis appears to become more echogenic in elderly people. CONCLUSION: Much remains to be done if we are to reach consensus on the effects of age on skin structure and function. Future studies would be benefited by increased standardization of skin sites tested, methodology, and increased sample size. PMID- 16221139 TI - Pigment distribution in melanocytic lesion images: a digital parameter to be employed for computer-aided diagnosis. AB - BACKGROUND/PURPOSE: Since in early melanoma (MM) and especially in in situ MM differential structures, which are diagnostic for MM may be lacking, pigment distribution asymmetry represents an important diagnostic feature. Our aim was to automatically assess pigment distribution in images referring to MMs, atypical nevi (AN) and clearly benign nevi (BN), and to evaluate the diagnostic capability of numerical parameters describing a non homogeneous distribution of pigmentation. METHODS: An image analysis program enabling the numerical assessment of pigment distribution in melanocytic lesions (ML), based on evaluation and comparison of red, green, blue (RGB) colour components inside image colour blocks, was employed on 459 videomicroscopic digital images, referring to 95 MMs, 76 AN and 288 BN. RESULTS: Significant differences in pigment distribution parameters (mean RGB distance, variance and maximum distance) between the three ML populations were observed, permitting a good discrimination of MMs. On the test set comprising 230 lesion images, the area under the curve value of the receiver operating characteristic curve was 0.933. For a D score equal to 0, corresponding to the best diagnostic accuracy (86.6%), a sensitivity of 87.5% and a specificity of 85.7% were obtained. CONCLUSION: This original evaluation method for digital pigment distribution, based on mathematical description and comparison of colours in different image blocks, provides numerical parameters to be implemented in image analysis programs for computer-aided MM diagnosis. PMID- 16221140 TI - Practical color calibration for dermoscopy, applied to a digital epiluminescence microscope. AB - BACKGROUND/PURPOSE: The assessment of colors is essential for melanoma (MM) diagnosis, both for pattern analysis on dermoscopic images, and when using semiquantitative methods. Our aim was to provide a simple, precise characterization and reproducible calibration of the color response for dermoscopic instruments. METHODS: Three processes were used to correct the non uniform illumination pattern of the instrument, to easily estimate the camera gamma settings and to describe the color space conversion matrices required to produce standard images, in any color space. A specific color space was also developed to optimize the representation of dermatoscopic colors. The calibration technique was tested both on synthetic reference surfaces and on real images by comparing the difference between the images colors obtained with two different equipments. RESULTS: The differences between the images acquired by means of the two instruments, calculated on the reference patterns after calibration, were up to 10 times lower then before, while comparison of histograms referring to real images provided an improvement of about seven times on average. CONCLUSIONS: A complete workflow for dermatologic image calibration, which allows the user to continue using his own software and algorithms, but with a much higher informative content, is presented. The technique is simple and may improve cooperation between different research centers, in teleconsulting contexts or for result comparisons. PMID- 16221141 TI - Digital image enhancement for in vivo laser scanning microscopy. AB - BACKGROUND/PURPOSE: In vivo confocal laser scanning microscopy (CLSM) is a new method that provides skin images in horizontal plane at a level of resolution that allows to view microanatomic structures. This study examines whether certain digital image-processing steps can increase the visibility of various structures in CLSM. STUDY DESIGN: Fifty images were taken from normal skin of 25 probands, and 39 image enhancement procedures were created. Eight procedures that seemed to provide some quality enhancement were deliberately selected for further evaluation. Subsequently, a collection of random pairs of the original image and an image submitted to any of the eight selected procedures was rated by five independent observers. RESULTS: In three of the eight procedures tested, the modified image was significantly preferred to the original image (chi2-test,: P< or =0.001). In particular, smoothing, shading correction, delineate and grey level normalization in various combinations were helpful in showing the characteristic honeycomb pattern, pigmented basal cell layer, cell borders and the nuclei more clearly. CONCLUSION: Digital image processing may help to increase visibility of in vivo CLSM images. PMID- 16221143 TI - Skin function parameters in intensive-care patients. AB - BACKGROUND/PURPOSE: The study was performed to investigate the transepidermal water loss (TEWL) and pH-value in patients in intensive care. METHODS: Forty intensive-care patients (22 men, 18 women) were included in the study. TEWL and pH-values were measured at admission, and after 24, 96 and 168 h. The areas of measurement were the forehead, the volar forearm, paraumbilical and the ventral thigh. The measurements were made under standardized environmental conditions according to the recommendations of the EMCO Group. RESULTS: Elevated values were found on the forehead compared with the other skin areas examined. There was no significant change in mean TEWL-values in any patient in the course of the study. There was also no significant influence of TEWL at the time of admission on the prognosis. The course analysis of the mean pH-values, however, showed that patients who developed a systemic inflammatory response syndrome (SIRS) or sepsis during the further course had a higher pH-value over the entire study period. CONCLUSION: TEWL and the pH of the skin surface could be measured at bedside in the intensive-care unit and delivered reproducible results. These parameters appear, however, to be relevant only for subgroups of patients under intensive care. PMID- 16221142 TI - Fluorescence fibre-optic confocal microscopy of skin in vivo: microscope and fluorophores. AB - BACKGROUND/AIMS: Fibre-optic confocal imaging in vivo is a new approach in the assessment of human skin. The objective is to describe a novel instrument and its operation and use in combination with fluorophores. METHODS: The Stratum is a fibre-optic fluorescence confocal microscope especially developed for the study of skin and mucous membranes. The system is flexible and any body site can be studied with a hand-held scanner. The light source is a 488 nm argon ion laser. Horizontal (en face) images of the epidermis and outer dermis are produced with cellular resolution. Magnification is approximately 1000 x . Fluorescein sodium is routinely used as fluorophore (intradermal injection or application to the skin surface). This fluorophore is safe for human use in vivo, but other substances (rhodamine B, Acridine Orange, green fluorescent protein, curcumin) have also been studied. RESULTS: The instrument produces sharp images of epidermal cell layers from the epidermal surface to the sub-papillary dermis, with sub-cellular resolution. The scanner is flexible in use. The technique of intradermal fluorophore injection requires some skill. CONCLUSIONS: We consider this fibre-optic instrument a potentially important tool in skin research for non invasive optical biopsy of primarily the epidermis. Present use is focussed on research applications, where the fluorophore distribution in the skin may illustrate morphological changes in the epidermis. PMID- 16221144 TI - Objective evaluation for severity of atopic dermatitis by morphologic study of skin surface contours. AB - BACKGROUND/AIMS: Wide variation in outcome methodology can make the interpretation of patient outcomes confusing and the comparison of the results of different studies almost impossible. It is important to objectively measure and record the severity of atopic dermatitis (AD) for routine clinical practice and research. The aim of this study was to evaluate whether morphologic study of skin surface contours might be helpful to objectively quantify the severity of AD. METHODS: Thirty atopic patients (12 females, 18 males) participated in this study. Moisturizer was applied twice daily for 2 weeks. Bioengineering methods such as D-Squame, corneometer, evaporimeter, and spectrophotometer were measured at the start of the study and after 1 week and 2 weeks. In addition, we assessed moisturizer effects after 3 h of moisturizer application.The stereoimage optical topometer (SOT) based on a new concept of stereoimage was applied for this study. We compared SOT, other bioengineering methods, and the severity scoring of atopic dermatitis (SCORAD) index. RESULTS: After 3 h of application with moisturizer, the results measured by SOT, conventional optical profilometer (COP), D-Squame, and corneometer showed significant differences (P<0.05). After 1 and 2 weeks, there were significant changes in the results measured by SOT, COP, D-Squame, corneometer, spectrophotometer, and SCORAD index. We observed a significant correlation between bioengineering methods and the SCORAD index (P<0.05). CONCLUSION: These data indicate that morphologic study of skin surface contours are useful in evaluating of AD severity. If we would combine methods to evaluate the physiologic changes and those such as SOT to measure the morphological changes of skin surface, we could evaluate more objectively and quantitatively the severity of AD. PMID- 16221145 TI - Non-invasive and microinvasive electrical impedance spectra of skin cancer - a comparison between two techniques. AB - BACKGROUND/PURPOSE: Bio-electrical impedance spectra of skin cancer and other lesions can be assessed using both regular non-invasive probes and a novel type of microinvasive electrode system with a surface furnished with tiny spikes that penetrate stratum corneum. The aim of the study was to compare the accuracy of detection for various types of skin cancer using impedance spectra measured with these two different electrode systems in an objective way without optimising the power of discrimination. METHODS: Impedance spectra of 99 benign nevi, 28 basal cell carcinomas (BCC), and 13 malignant melanomas (MM) were measured using the two electrode systems. Classification of the lesions was made using Fisher's linear discriminant, and the sensitivities and specificities of the techniques were estimated using cross-validation. RESULTS: The best separation between nevi and BCC was achieved using the regular non-invasive probe (96% sensitivity and 86% specificity), whereas the best separation between nevi and MM was achieved using the microinvasive electrodes (92% sensitivity and 80% specificity). CONCLUSIONS: Our results indicate that the choice of electrode system is application dependent. PMID- 16221146 TI - Evaluation of nonlinear elastic behaviors of skin. PMID- 16221150 TI - Darwinian competition and the pathogenesis of opportunistic infection in the transplant recipient. PMID- 16221151 TI - Impairment of renal function after islet transplant alone or islet-after-kidney transplantation using a sirolimus/tacrolimus-based immunosuppressive regimen. AB - The immunosuppressive (IS) regimen based on sirolimus/low-dose tacrolimus is considered a major determinant of success of the Edmonton protocol. This regimen is generally considered safe or even protective for the kidney. Herein, we analyzed the impact of the sirolimus/low-dose tacrolimus combination on kidney function. The medical charts of islet transplant recipients with at least 6 months follow up were reviewed. There were five islet-after-kidney and five islet transplantation alone patients. Serum creatinin, albuminuria, metabolic control markers and graft function were analyzed. Impairment of kidney function was observed in six of 10 patients. Neither metabolic markers nor IS drugs levels were significantly associated with the decrease of kidney function. Although a specific etiology was not identified, some subsets of patients presented a higher risk for decline of kidney function. Low creatinin clearance, albuminuria and long-established kidney graft were associated with poorer outcome. PMID- 16221152 TI - Obesity increases mortality in liver transplantation--the Danish experience. AB - Obesity is increasing in the western world at an epidemic rate. The USA results in obese patients undergoing orthotopic liver transplantation (OLT) are divergent, and so far no European experience has been reported. This study was designed to determine if obesity is a risk factor for mortality and morbidity in OLT in a medium-size European center. In a retrospective study of the records of 365 consecutive patients who had undergone OLT from 1990 to 2003, 20 obese patients [body mass index (BMI) > 30 kg/m2] were identified. Their data were compared with those of the nonobese (BMI < 30 kg/m2) patients operated immediately before. There were no differences in demographic data, diagnosis leading to OLT, United Network of Organ Sharing (UNOS) classification, Child-Pugh score, or preoperative morbidity. The groups were also comparable concerning donor data, duration of operation, use of blood products, intensive care unit (ICU), or hospital admission. Mortality was, however, significantly increased in the obese group (P = 0.01). Our study clearly demonstrates an increased mortality in obese patients undergoing OLT and the relative scarcity of organs taken into account, it seems reasonable to consider obesity as a relative contraindication to OLT. PMID- 16221153 TI - Blood salvage autotransfusion during transplantation for hepatocarcinoma: does it increase the risk of neoplastic recurrence? AB - Impact of intraoperative blood salvage autotransfusion (IBSA) on neoplastic recurrence. during liver transplantations for hepatocellular carcinoma (LT-HCC). Between January 1989 and February 2003, 16 patients received a LT-HCC without IBSA. This group was compared with 31 patients who received the same surgical procedure during the same period, but with IBSA. Data were prospectively collected. All patients had at least a 1-year postoperative follow up. Pairing was made according to the size of the largest nodule. The percentage of recurrence observed in the two groups was similar: 6.4% in the IBSA group vs. 6.3% in the group without IBSA. The median amount of transfused salvage blood was 1558 ml. The differences observed between the two groups concerned the Child score which was A in 58% patients of the IBSA group vs. 80% in the other group; the percentage of severe portal hypertension was 55% in the IBSA group vs. 31%; the median number of packed red blood cell units transfused intraoperatively was 7 in the IBSA group vs. 0, and the median number of frozen fresh plasma units transfused intraoperatively was 11 in the IBSA group vs. 4.5. It appears that IBSA, essentially used during the most haemorrhagic transplantations, could be used in the case of HCC because it does not modify the risk of neoplastic recurrence. PMID- 16221155 TI - Predicting mortality after kidney transplantation: a clinical tool. AB - An increasing number of patients referred for transplantation are older and have complex comorbidity affecting outcome. Patient counseling is often empiric and time consuming. For the physician there are few clinical tools available to help quantify survival chances after transplantation. We used registry data to develop a series of tables that could be used in the clinical setting to predict survival probability. Using data from the Canadian Organ Replacement Registry, we generated clinical survival tables using Cox's regression model. Model covariates included age, race, gender, treatment period, primary renal disease cause, donor source, months on dialysis and comorbidities. A total of 6324 patients were included, 22% had > or =1 comorbid condition at baseline. After adjustment for age, gender and cause of renal disease, increased comorbidity was strongly associated with reduced patient-survival (P < 0.05). Age and comorbidity specific clinical survival tables showing the expected 1-, 3- and 5-year patient survival probabilities were generated. Separate tables were created for diabetics, nondiabetics, living-donor organs and deceased-donor transplantation. Patient specific survival data can be estimated from registry data. We suggest annual or biannual tables generated by national registries across Europe and N. America, may be useful to those physicians faced with counseling patients and families. PMID- 16221154 TI - In adult-to-adult living donor liver transplantation hepaticojejunostomy shows a better long-term outcome than duct-to-duct anastomosis. AB - Roux-en-Y hepaticojejunostomy (RYHJ) has been the standard biliary reconstruction in adult-to-adult living donor liver transplantation (ALDLT). Recently, duct-to duct anastomosis (DD) has been introduced. This study compared the outcomes of RYHJ and DD. For 4 years, 74 recipients underwent ALDLT and were followed up for at least 2 years. The patients were divided into three groups, RYHJ group (n = 18), DD with a stent (DD + S) group (n = 35), and DD without a stent (DD - S) group (n = 21). Overall, biliary complications were developed in 32.4% patients. The biliary complication rate was 11.1%, 48.5% and 33.3% in RYHJ, DD + S and DD - S groups, respectively (P = 0.047). Bile leaks occurred in 28.5% of DD + S group. The incidence of biliary stricture was 5.3%, 20.2% and 28.6% in RYHJ, DD + S and DD - S group, respectively. Most complications (83.3%) were resolved nonsurgically. RYHJ has a better long-term outcome than DD in ALDLT. Subgroup analysis of DD group showed that DD - S group had no bile leaks, but still had a higher incidence of bile duct strictures. However, because this study was a retrospective review there are limitations in analyzing the data and confirming the conclusion. A randomized-prospective study will be needed to confirm these findings. PMID- 16221157 TI - Effect of donor age on the outcome of living-related kidney transplantation. AB - The study compared the results of kidney transplantation from living-related donors older and younger than 60 years. The 273 kidney graft recipients were divided into group 1 (115 recipients of older grafts) and group 2 (158 recipients of younger grafts). The frequency of acute rejection (AR) episodes was similar in both groups but slow graft function occurred more frequently in group 1. The frequency of chronic renal allograft dysfunction in the first post-transplant year was significantly higher in group 1 than in group 2. Patient and graft survival was significantly worse in group 1. Risk factors for graft loss were the difference between donor and recipient age and AR. Donor age and graft function were risk factors for patient death. Although kidneys from older donors provide a statistically poorer transplant outcome, they are clinically acceptable, especially when waiting time is prolonged and access to dialysis limited. PMID- 16221156 TI - Recipient levels and function of von Willebrand factor prior to liver transplantation and its consumption in the course of grafting correlate with hepatocellular damage and outcome. AB - Von Willebrand factor (vWF) is a major platelet adhesion molecule at sites of vascular injury, such as observed in ischemia/reperfusion injury following orthotopic liver transplantation (OLT). Thirty-three OLT patients were divided into groups with elevated or low markers of hepatocellular damage (high and low HD). Whole-blood aggregometry was performed to evaluate platelet function. Multimeric analysis was utilized to evaluate functional vWF levels in the course of OLT. Donor and recipient demographics were comparable among groups. Low-HD patients demonstrated better preserved coagulation parameters on POD 1-6 if contrasted to high-HD patients. One year graft survival for the high-HD group was lower than low-HD patients (P = 0.037). Preoperative vWF-dependent platelet aggregation and functional vWF plasma levels correlated directly with alanine transaminase levels early after OLT as did the decrease of functional vWF to reperfusion. In summary, these data suggest that vWF may serve as a significant mediator of platelet recruitment and hepatocellular injury in the graft following reperfusion. PMID- 16221158 TI - Opting-out systems: no guarantee for higher donation rates. AB - There are considerable differences in the number of organ donations between countries. It is assumed that opting-out systems have a significantly positive impact on the national organ donation rate. The aim of our study was to establish whether different consent systems explain the difference in organ donation rates between countries when taking into account the difference in relevant mortality rates. For this study, we compared data on donation and relevant mortality rates for 10 different countries as well as information on the existing consent systems. This international comparative study shows that there is a strong correlation between mortality rates and donation rates (Spearman's rho = 0.81 (P < 0.01). International comparative legal research has shown that the differences between decision systems are marginal. When the national organ donation rates are corrected for mortality rates, these findings are confirmed: the donor efficiency rate shows that opting-out systems do not automatically guarantee higher donation rates than opting-in systems. PMID- 16221159 TI - Incidence and prognosis of cancer following heart transplantation using RATG induction therapy. AB - Cancer limits survival following heart transplantation. The study's objectives were to evaluate the incidence and risk factors for cancers after heart transplantation and to assess the association between i.v. thymoglobuline induction therapy [rabbit antithymocyte immunoglobulin, (RATG)] and neoplasia. From 1982 to 2002, prospective data were gathered for 207 heart transplant recipients. Except from 1982 to 1987, all patients received a 3-day course of i.v. RATG following transplantation. Forty-three malignant neoplasms (21%) were diagnosed. The most common were: skin (42%), lung (12%), prostate (9%), genitourinary (9%) and lymphoma (5%). Mean length of follow-up after transplantation was 99 +/- 57 months. Mean survival after diagnosis was 52 +/- 44 months. Multivariate analysis showed no significant increase in the incidence of cancer with recipient age, sex, number of rejection episodes, the type of immunosuppression or the use of RATG. Patients receiving RATG developed their malignancies significantly earlier after transplantation (P =0.007) and succumbed faster after the diagnosis (P = 0.06). Cancer is a limiting event for long-term survival after heart transplantation. No individual risk factors allow predicting its development. In the present cohort, RATG does not have carcinogenic effects following transplantation, but is associated with a more precocious development of malignancies. PMID- 16221160 TI - Humoral immune response after kidney transplantation is enhanced by acute rejection and urological obstruction and is down-regulated by mycophenolate mofetil treatment. AB - The anti-allograft immune response may have a cellular and a humoral component. Lymphocytotoxic antibodies (Ab) and anti-human leucocyte antigen (HLA) Ab present before kidney transplantation carry an enhanced risk of acute rejection. Current immunosuppressive drugs act predominantly upon the cellular immune pathway which may leave unopposed the humoral mechanisms of anti-allograft response. We studied the production of lymphocytotoxic Ab and anti-HLA Ab after kidney transplantation under different drug therapies. Two hundred and sixty-four consecutive kidney transplant recipients treated with different immunosuppressive drugs, either stable and or with previous acute rejection or acute urologic obstruction, entered this study. Lymphocytotoxic Ab and anti-HLA Ab were evaluated by complement-dependent cytotoxicity and by ELISA. Ab donor-specificity was determined by flow cytometry. Both lymphocytotoxic Ab and anti-HLA Ab were significantly increased in acute rejection whatever the immunosuppressive regimen and almost significantly in urologic obstruction treated with azathioprine (AZA) groups. The presence of antidonor-specific Ab was associated with a significantly higher rate of graft loss. Mycophenolate mofetil (MMF) therapy significantly down regulated Ab synthesis in all patients groups when compared with AZA. The development of humoral antidonor response post-transplantation is associated with a dismal graft prognosis. This is the first report that acute urologic obstruction may be followed by unspecific lymphocytotoxic and anti-HLA Ab synthesis, surmising that a protracted obstruction may promote renal fibrosis through antibody mediation. The significant down-regulation of the humoral response by MMF when compared with AZA may herald a lower risk to mount a chronic rejection process. PMID- 16221161 TI - Are preoperative patterns of alcohol consumption predictive of relapse after liver transplantation for alcoholic liver disease? AB - Predictive factors for alcoholic relapse after liver transplantation (LT) performed for alcoholic liver disease (ALD) have been assessed in numerous studies, often with contradictory results. The aim of the study was to assess pretransplantation alcohol consumption characteristics on alcoholic relapse after LT. Patients transplanted for ALD for at least 6 months were included. An anonymous questionnaire assessed socio-demographic characteristics, medical history, and alcohol consumption before and after LT. Relapse was defined as any alcohol use after LT. Severe relapse was defined by heavy drinking: more than 21 units/week for males and 14 units/week for females. A total of 61 patients were studied. The mean follow up after LT was 49 +/- 34 months. Alcoholic relapse occurred in 32 of 61 patients (52%) and severe relapse in eight of 61 patients (13%). Risk factors for severe relapse were: length of abstinence before LT (P = 0.0001), more than one alcohol withdrawal before LT (P = 0.001), alcohol dependence (P = 0.05), alcohol abuse in first relatives (P = 0.05), and younger age (P = 0.05). Information on previous alcohol consumption (dependence, number of withdrawals, family history) helps to predict severe relapse after LT in patients with ALD, allowing early awareness and specific postoperative care. PMID- 16221162 TI - A case of acute humoral rejection in liver transplantation: successful treatment with plasmapheresis and mycophenolate mofetil. AB - We present a case of a 23-year-old female who underwent orthotopic liver transplantation (OLTx) for biliary atresia, 22 years after a failed Kasai operation. Unusually, her postoperative course was complicated by severe acute humoral rejection. In this case report, we discuss her management as well as the role of plasmapheresis in treating allograft dysfunction secondary to acute humoral rejection in liver transplant patients. PMID- 16221164 TI - Polyomavirus interstitial nephritis and concurrent post-transplant lymphoma in a renal allograft: coincidence or more? AB - We describe a case of an Epstein-Barr virus (EBV)-negative post-transplant large B-cell non-Hodgkin lymphoma located in the renal allograft, spleen, liver and left inguinal lymph node of a renal recipient and accompanied by a simultaneous polyomavirus-associated nephropathy. To our knowledge, this is the first report of a simultaneous polyomavirus infection and post-transplant lymphoproliferative disorder. PMID- 16221163 TI - Sirolimus-associated hepatotoxicity in the kidney graft recipient. AB - The aim of our paper was to describe hepatotoxicity of sirolimus (SRL) in a kidney graft recipient. We report the case of a 30-year-old male after kidney transplantation, treated with steroids, cyclosporin A and SRL, with steroid resistant acute rejection in anamnesis. At 16th month after transplantation, elevation of serum aminotransfereases was observed. After exclusion of common reasons of this condition, liver biopsy was performed. Nonspecific changes were observed, with probability of drug-induced injury. SRL was changed to mycophenolate mofetil, which was followed by quick normalization of serum aminotransferase levels. Hepatoxicity is a rare complication of SRL therapy and may be connected with some diagnostic and/or therapeutic problems. Conversion to another immunosuppressant seems to be an appropriate procedure in this condition. PMID- 16221167 TI - Neural stem cells and regeneration of injured spinal cord. AB - Recent progress in the stem cell biology has led much insight into new therapeutic interventions aiming for the regeneration of the damaged central nervous system. The major strategies can be classified into two subgroups: (1) activation of endogenous neural stem cells, and (2) cell transplantation therapies. In either of these strategies, it is crucial to understand the underlying mechanisms of maintenance, activation, and differentiation of neural stem cells and subsequent process, including the migration, survival, and functional maturation of differentiated cells. In this paper, we would like to summarize our recent findings on the therapeutic interventions of the injured spinal cord, especially focusing on the development of treatment for the acute phase of spinal cord injury with anti-interleukin (IL)-6 receptor blocking antibody. PMID- 16221168 TI - Stem cells and progenitor cells in renal disease. AB - Stem cells and progenitor cells are necessary for repair and regeneration of injured renal tissue. Infiltrating or resident stem cells can contribute to the replacement of lost or damaged tissue. However, the regulation of circulating progenitor cells is not well understood. We have analyzed the effects of erythropoietin on circulating progenitor cells and found that low levels of erythropoietin induce mobilization and differentiation of endothelial progenitor cells. In an animal model of 5/6 nephrectomy we could demonstrate that erythropoietin ameliorates tissue injury. Full regeneration of renal tissue demands the existence of stem cells and an adequate local "milieu," a so-called stem cell niche. We have previously described a stem cell niche in the kidneys of the dogfish, Squalus acanthus. Further analysis revealed that in the regenerating zone of the shark kidney, stem cells exist that can be induced by loss of renal tissue to form new glomeruli. Such animal models improve our understanding of stem cell behavior in the kidney and may eventually contribute to novel therapies. PMID- 16221169 TI - Stem cell therapy for human brain disorders. AB - Transplantation of stem cells or their derivatives, and mobilization of endogenous stem cells in the adult brain, have been proposed as future therapies for various brain disorders such as Parkinson's disease and stroke. In support, recent progress shows that neurons suitable for transplantation can be generated from stem cells in culture, and that the adult brain produces new neurons from its own stem cells in response to injury. However, from a clinical perspective, the development of stem cell-based therapies for brain diseases is still at an early stage. Many basic issues remain to be solved and we need to move forward with caution and avoid scientifically ill-founded trials in patients. We do not know the best stem cell source, and research on embryonic stem cells and stem cells from embryonic or adult brain or from other tissues should therefore be performed in parallel. We need to understand how to control stem cell proliferation and differentiation into specific cell types, induce their integration into neural networks, and optimize the functional recovery in animal models closely resembling the human disease. All these scientific efforts are clearly justified because, for the first time, there is now real hope that we in the future can offer patients with currently intractable diseases effective cell based treatments to restore brain function. PMID- 16221170 TI - Mesenchymal, but not hematopoietic, stem cells can be mobilized and differentiate into cardiomyocytes after myocardial infarction in mice. AB - Bone marrow (BM) cells are reported to contribute to the process of regeneration following myocardial infarction. The present study examined two independent clonal studies to determine the origin of bone marrow (BM)-derived cardiomyocytes. First, we transplanted single CD34(-)c-kit(+)Sca-1(+)lineage(-) side population cells (hematopoietic stem cells) from enhanced green fluorescent protein (EGFP)-transgenic mice into lethally irradiated mice, induced myocardial infarction, and treated them with G-CSF to mobilize stem cells. At 8 weeks, we could not find any EGFP(+) cardiomyocytes. In contrast, more than 5000 EGFP(+) cardiomyocytes were observed in whole BM cell-transplanted mice, suggesting that they were derived from non-hematopoietic cells. Next, clonally purified mesenchymal stem cells (MSC) that expressed EGFP in the cardiomyocyte-specific manner were transplanted directly into BM of lethally irradiated mice, and similar experiment was performed. EGFP(+) actinin(+) cells were observed in the ischemic myocardium, indicating that MSC had been mobilized and differentiated into cardiomyocytes. Together, these results suggest that the origin of the BM derived cardiomyocytes is MSC. PMID- 16221171 TI - Roles of HNF-1beta in kidney development and congenital cystic diseases. AB - Hepatocyte nuclear factor-1beta (HNF-1beta) is a Pit-1/Oct-1/Unc-86 (POU)/homeodomain-containing transcription factor that regulates tissue-specific gene expression in the kidney, liver, pancreas, and other epithelial organs. Mutations of HNF-1beta produce maturity-onset diabetes of the young type 5 (MODY5) and are associated with congenital cystic abnormalities of the kidney. Transgenic mice expressing mutant HNF-1beta under the control of a kidney specific promoter develop kidney cysts and renal failure, which is similar to the phenotype of humans with MODY5. Similarly, kidney-specific deletion of HNF-1beta using Cre/loxP recombination results in renal cyst formation. HNF-1beta directly regulates the Pkhd1 promoter. HNF-1beta mutant mice show decreased expression of Pkhd1, the gene that is mutated in humans with autosomal-recessive polycystic kidney disease (ARPKD). These studies demonstrate that HNF-1beta is required for the development of the mammalian kidney. They establish a previously unrecognized link between two renal cystic diseases, MODY5 and ARPKD, and suggest that the mechanism of cyst formation in humans with mutations of HNF-1beta involves down regulation of PKHD1 gene transcription. PMID- 16221173 TI - The role of Notch signaling in specification of podocyte and proximal tubules within the developing mouse kidney. AB - Notch genes encode transmembrane receptors that mediate intercellular interaction by binding to the ligands on the adjacent cells. Due to early embryonic lethality in mice deficient for some Notch pathway genes, the role of Notch signaling for kidney development has not yet been defined. Using an antibody specific to the N terminal end of gamma-secretase-cleaved Notch 1, we found evidence for Notch 1 activation in the comma-shaped and S-shaped bodies. We therefore cultured embryonic (E) day E12.5 mouse metanephroi in the presence of a gamma-secretase inhibitor, N-S-phenyl-glycine-t-butyl ester (DAPT), to block Notch signaling. Fewer renal epithelial structures were observed, with a severe deficiency in proximal tubules and glomerular podocytes. Distal tubules were present but at a reduced number, and this was accompanied by an increase in intervening, nonepithelial cells. By culturing day E14.5 metanephroi, we observed the formation of podocyte clusters after 3 days of DAPT treatment. These observations suggest that gamma-secretase activity, probably through activation of Notch, is not essential for podocyte formation beyond the stage of S-shaped body but is required for the proximal tubule and podocyte fates when S-shaped bodies are forming. PMID- 16221172 TI - Essential roles of Sall1 in kidney development. AB - SALL1 is a mammalian homologue of the Drosophila region-specific homeotic gene spalt (sal) and heterozygous mutations in SALL1 in humans lead to Townes-Brocks syndrome. We isolated a mouse homologue of SALL1 (Sall1) and found that mice deficient in Sall1 die in the perinatal period with kidney agenesis. Sall1 is expressed in the metanephric mesenchyme surrounding ureteric bud and homozygous deletion of Sall1 results in an incomplete ureteric bud outgrowth. Therefore, Sall1 is essential for ureteric bud invasion, the initial key step for metanephros development. We also generated mice in which a green fluorescent protein (GFP) gene was inserted into the Sall1 locus and we isolated the GFP positive population from embryonic kidneys of these mice by fluorescence activated cell sorting (FACS). We then compared gene expression profiles in the GFP-positive and -negative population using microarray analysis, followed by in situ hybridization. We detected many genes known to be important for metanephros development, and genes expressed abundantly in the metanephric mesenchyme. We also found groups of genes which are not known to be expressed in the metanephric mesenchyme. Thus a combination of microarray technology and Sall1-GFP mice is useful for systematic identification of genes expressed in the developing kidney. PMID- 16221174 TI - Organogenesis of the endocrine pancreas. AB - Organogenesis of the endocrine pancreas. Embryonic pancreatic primordia transplanted into diabetic animal hosts undergo selective endocrine differentiation in situ and normalize glucose tolerance. Pancreatic primordia can be transplanted across isogeneic, allogeneic, and both concordant (rat to mouse) and highly disparate (pig to rodent) xenogeneic barriers. This review explores the therapeutic potential for pancreatic organogenesis posttransplantation of embryonic primordia. PMID- 16221175 TI - Kidney tubular epithelium is restored without replacement with bone marrow derived cells during repair after ischemic injury. AB - The kidney has the ability to restore the structural and functional integrity of the proximal tubule, which undergoes extensive epithelial cell death after prolonged exposure to ischemia. In order to study the role that adult bone marrow derived stem cells might play in kidney remodeling after injury, we employed a murine model of ischemia/reperfusion (I/R) injury in which the degree of injury, dysfunction, repair, tubular cell proliferation and functional recovery have been characterized [Park KM, et al, J Biol Chem 276:11870-11876, 2001]. We generated chimeric mice using marrow from mice expressing the bacterial LacZ gene, or the enhanced green fluorescence protein (eGFP) gene, or from male mice transplanted into female mice. The establishment of chimerism was confirmed at 6 weeks following transplantation in each case. I/R injury was induced in chimeric mice by occluding the renal arteries and veins with microaneurysm clamps for 30 minutes. After functional recovery in the eGFP chimeras, although there were many interstitial cells, no tubular cells were derived from bone marrow cells. In the bacterial beta-galactosidase (beta-gal) chimeric mice we found evidence of mammalian (endogenous) beta-gal by 5-bomo-4-chloro-3-indolyl-beta-D galactopyranoside (X-gal) staining, but not bacterial beta-gal in tubule cells. Detection of the Y chromosome by fluorescence in situ hybridization (FISH) in the postischemic kidneys of gender-mismatched chimeras revealed Y chromosome positivity only in the nuclei of interstitial cells, when scrutinized by deconvolution microscopy. In our model of I/R injury there was a large amount of proliferation of surviving, injured tubular cells indicating that the injured tubule is repopulated by daughter cells of surviving tubular cells. Analysis of the phenotype of interstitial and vascular cells following I/R injury revealed small numbers of peritubular endothelial cells to be derived from bone marrow cells that may serve in the repair process. PMID- 16221197 TI - Vitamin D and its analogues: do they protect against cardiovascular disease in patients with kidney disease? AB - BACKGROUND: Patients with chronic kidney disease (CKD) are at high risk for cardiovascular disease, and despite recent advances in hypertension control, anemia management, and dialysis adequacy, mortality remains high. Improved understanding of nontraditional risk factors, including those present at early phases in CKD, may lead to novel therapeutic strategies. CKD has been demonstrated to be an independent risk factor for cardiovascular disease in the general population, but data are lacking as to the associated potential abnormalities that occur in association with reduced glomerular filtration rate (GFR), which may contribute to this increased risk. Data are accumulating regarding the role of abnormalities of calcium, phosphorus, vitamin D, and parathyroid hormone (PTH) in cardiovascular disease. Vitamin D deficiency is present even in the early stages of CKD. Vitamin D plays a central role in calcium-phosphorus homeostasis, regulation of PTH, and formation and maintenance of bone. However, until recently, vitamin D has not been considered to have a biologic role in CKD beyond mineral regulation, or has been considered as a negative factor contributing to soft tissue and cardiovascular calcification. In light of recent observational studies showing an association of vitamin D therapy and survival benefit in hemodialysis patients, the effects of vitamin D on cardiovascular system have become a heavily debated issue. METHODS: A Medline search was performed to identify relevant literature describing the role of vitamin D in the pathogenesis of cardiovascular disease. Both the experimental and clinical literatures in English were reviewed. RESULTS: The accumulating published data demonstrate both associative relationships and mechanisms for biologic plausibility. The following three potential mechanisms may be important for the protective effects of vitamin D against cardiovascular disease mortality: vitamin D can inhibit various aspects of inflammation, which have been established as a key pathogenic mechanism in atherosclerosis; vitamin D exerts an antiproliferative effect on myocardial cell hypertrophy and proliferation, which underlies the pathogenesis of congestive heart failure; and vitamin D acts as a negative endocrine regulator for the renin-angiotensin system, which itself plays an important independent role in hypertension and cardiovascular health. CONCLUSION: Vitamin D deficiency might be an underestimated nonclassical risk factor for cardiovascular disease in CKD. Based on a review of the evidence, from both basic science and clinical studies, this article supports the possible protective role of vitamin D beyond its effect on mineral metabolism, and suggests the need for ongoing evaluation of the role of vitamin D in cardiovascular health in the CKD population. PMID- 16221198 TI - Evolving concepts in the quantitative analysis of the determinants of the plasma water sodium concentration and the pathophysiology and treatment of the dysnatremias. AB - The physiologic and clinical implications of the empirical formula originally discovered by Edelman et al [J Clin Invest 37:1236-1256, 1958] relating the plasma water sodium concentration ([Na(+)](pw)) to the total exchangeable sodium (Na(e)), total exchangeable potassium (K(e)), and total body water (TBW) have recently been elucidated. It is quite remarkable that the full significance of the Edelman equation discovered almost 50 years ago had remained unrecognized by clinicians and physiologists until recently. Although Edelman and colleagues had shown that the [Na(+)](pw) is proportional to the magnitude of (Na(e)+ K(e))/TBW, the linear equation relating [Na(+)](pw) to (Na(e)+ K(e))/TBW had a slope greater than unity of 1.11, and a non-zero y intercept of -25.6 whose significance was unrecognized and more often than not ignored. It has recently been demonstrated that the slope and y intercept in this equation are quantitatively determined by several additional physiologic parameters, which in addition to (Na(e)+ K(e))/TBW, play a role both in modulating the [Na(+)](pw) and in the generation of the dysnatremias. Even more remarkably, based only on the theoretical principles of Gibbs-Donnan and osmotic equilibrium, all the physiologic parameters that determine the magnitude of the [Na(+)](pw) can be incorporated into a simple conceptual and mathematical framework that sheds light on a broad of range of seemingly unrelated topics that have heretofore been treated separately clinically, including (1) effect of changes in the mass balance of Na(+), K(+), and H(2)O on the [Na(+)](pw); (2) modulation of [Na(+)](pw) in hyperglycemic states; (3) definition of an isonatric solution; (4) current formulas used to quantitate electrolyte-free water excretion; (5) complex role of K(+) in modulating the [Na(+)](pw); and (6) quantitative analysis of the generation and treatment of the dysnatremias. Moreover, this analysis has also proven to be an indispensable tool for deriving new formulas to aid the clinician in both interpreting the pathogenesis and treating the dysnatremias. PMID- 16221199 TI - Hereditary amyloidosis in early childhood associated with a novel insertion deletion (indel) in the fibrinogen Aalpha chain gene. AB - BACKGROUND: Systemic amyloidosis occurring in early childhood is extremely rare, and is usually of AA type complicating chronic inflammatory diseases. We report the molecular basis of amyloidosis in a Korean girl who presented at 7 years of age with asymptomatic proteinuria and developed amyloid hepatomegaly and end stage renal failure within 2 years. METHODS: Renal biopsy showed enlarged glomeruli virtually replaced by amyloid, but without interstitial or vascular involvement. The histologic appearance was identical to that seen in patients with hereditary fibrinogen Aalpha chain Glu526Val amyloidosis, and the amyloid deposits stained specifically with antibodies to fibrinogen. Mutations were sought in the genes of the amyloidogenic proteins, transthyretin, apolipoprotein AI, lysozyme and fibrinogen Aalpha chain genes by polymerase chain reaction (PCR) and sequencing. RESULTS: A unique frameshift insertion-deletion (indel) mutation was identified in one allele of her fibrinogen Aalpha chain gene, which encodes a partly novel peptide and a premature stop signal, similar to the two previously reported amyloidogenic point deletions at codons 522 and 524 in this molecule. The mutation was absent in samples verified to be from her parents, indicating that it had occurred de novo. CONCLUSION: This is the first description of hereditary fibrinogen Aalpha chain amyloidosis in an Asian individual, and the distinctive renal histology offered a strong clue to the diagnosis. The disease is potentially curable by combined hepatorenal transplantation. PMID- 16221200 TI - The C-terminal tail of aquaporin-2 determines apical trafficking. AB - BACKGROUND: Aquaporin-2 (AQP-2) proteins are mainly expressed at the apical region of the collecting duct cells. We previously reported three different mutations in the C-terminus of AQP-2 that all-cause autosomal-dominant nephrogenic diabetes insipidus. When one of these mutant AQP-2s was expressed in Madin-Darby canine kidney (MDCK) cells, it was mistargeted to the basolateral membrane, suggesting a critical role of the C-terminal tail in the apical trafficking of AQP-2. METHODS: Portions of the AQP-2 C-terminal tail (residues 226-271) were mutated by the polymerase chain reaction (PCR) technique and inserted into the pcDNA3.1 vector. Constructs were transfected into MDCK cells to examine the localization of mutated AQP-2 proteins by immunofluorescence microscopy. Cell surface expression was detected by biotinylation assay. RESULTS: The wild-type AQP-2 was localized at the apical membrane, whereas mutants lacking residues 262-271 (the last 10 amino acids) were predominantly distributed in the endoplasmic reticulum. Deletion mutants of the initial (226-240del) and middle (241-252del) portions of the C-terminal tail were identified at the apical membrane, suggesting that residues 226-252 have no involvement in apical targeting. An AQP-4-AQP-2 chimera in which a portion of the AQP-4 C-terminal tail was replaced by the corresponding site in AQP-2 (residues 256-271) was found at the apical membrane. The sequence of the last 4 amino acids of AQP-2 (G-T-K-A) corresponds to a PDZ-interacting motif. Our investigations identified a mutant of this portion mostly localized to the subapical region. Further, apical expression was found to be significantly decreased in mutants lacking a consensus sequence for cyclic adenosine monophosphate (cAMP)-dependent phosphorylation (residues 253 256). CONCLUSION: The sequence at 256-271 is sufficient for apical trafficking in AQP-2. The putative PDZ-interacting motif (G-T-K-A, residues 268-271) plays a key role in apical membrane expression. In addition, cAMP-dependent phosphorylation was found to be critical for apical targeting. PMID- 16221201 TI - A role for microfilament-based contraction in branching morphogenesis of the ureteric bud. AB - BACKGROUND: Branching morphogenesis of the ureteric bud/collecting duct epithelium is an important feature of kidney development. Recent work has identified many transcription factors and paracrine signaling molecules that regulate branching, but the physical mechanisms by which these signals act remain largely unknown. The actin cytoskeleton is a common component of mechanisms of morphogenesis. We have therefore studied the expression of, and requirement for actin filaments in the ureteric bud, a branching epithelium of the mammalian kidney. METHODS: Embryonic kidney rudiments were grown in organ culture. Actin expression in kidneys growing normally and those in which branching was inhibited was examined using labeled phalloidin. The morphogenetic effects of inhibiting actin organization and tension using cytochalasin D, butanedione monoxime, and Rho kinase ROCK inhibitors were assessed using immunofluorescence. RESULTS: F actin is expressed particularly strongly in the apical domains of cells at the tips of branching ureteric bud, but this expression depends on the bud actively growing and branching. Blocking the polymerization of actin using cytochalasin D inhibits ureteric bud branching reversibly, as does blocking myosin function using butadiene monoxime. Inhibiting the activation of ROCK, a known activator of myosin, with the drugs Y27632 or with H1152 inhibits the expression of strong actin bundles in the ureteric bud tips and inhibits ureteric bud branching without inhibiting other aspects of renal development. CONCLUSION: The formation of tension-bearing actin-myosin complexes is essential for branching morphogenesis in the developing kidney. PMID- 16221202 TI - Growth hormone promotes glomerular lipid accumulation in bGH mice. AB - BACKGROUND: Bovine growth hormone (bGH) transgenic mice develop progressive glomerulosclerosis and exhibit abnormalities in hepatic lipid metabolism. We have previously shown that growth hormone up-regulates the low-density lipoprotein (LDL) receptor and 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) in mouse mesangial cells. However, a role of lipid abnormalities in bGH kidney disease has not yet been demonstrated. METHODS: Groups of bGH mice (5 and 11 months old) presenting with, respectively, moderate and severe degrees of glomerulosclerosis were compared to age-matched controls. Neutral lipid content in kidney cortex was determined by oil red-O staining, serum cholesterol, and triglycerides by enzymatic assays, relative mRNA expression of LDL receptors, HMGR, and scavenger receptor by real-time reverse transcription-polymerase chain reaction (RT-PCR), and HMGR protein expression by immunoblotting. Two younger (5 and 12 weeks old) groups of mice were used to study scavenger receptor expression at earlier time points. RESULTS: Serum cholesterol was significantly increased in bGH mice at 5 months, but triglycerides were lower than control levels at both 5 and 11 months. Renal cortex HMGR expression was elevated at the mRNA but not at the protein level in the 11-month-old bGH group compared to controls. However, glomerular neutral lipid staining and scavenger receptor mRNA expression were markedly increased in all bGH mice, including those at 5 weeks of age compared to respective controls. CONCLUSION: The bGH mouse exhibits an increased mesangial lipid content and elevated scavenger receptor mRNA expression as early as at 5 weeks of age, suggesting that an increased kidney uptake of oxidized LDL could play a role in the development of glomerulosclerosis in this mouse model. PMID- 16221203 TI - Disturbed Ca2+-signaling by chloroacetaldehyde: a possible cause for chronic ifosfamide nephrotoxicity. AB - BACKGROUND: Renal damage following chemotherapy with ifosfamide is attributed to the metabolic activation of the drug and the generation of chloroacetaldehyde (CAA). Little is known about the mechanism by which CAA impairs renal function. In this study the effect of CAA on intracellular Ca(2+) homeostasis in human renal proximal tubule cells (RPTEC) in primary culture was investigated. METHODS: Intracellular Ca(2+) was measured using the Ca(2+)-sensitive dye fura-2. Cell viability was determined by protein content and cell number. Oncotic and apoptotic cell death was assayed using trypan blue exclusion, caspase-3 activity, and 4',6-diamino-2-phenylindole (DAPI) staining. RESULTS: CAA (1.5 to 150 micromol/L) induced sustained elevations of intracellular free calcium ([Ca(2+)](i)) from 75 +/- 3 nmol/L to maximal 151 +/- 6 nmol/L. This effect was dependent on extracellular Ca(2+), but not Ca(2+) entry. The rise in [Ca(2+)](i) mediated by CAA could be attributed to inhibition of Na(+)-dependent extrusion of intracellular Ca(2+), indicating an inhibitory action of CAA on Na(+)/Ca(2+) exchange. Modulation of protein kinase A (PKA), but not protein kinase C (PKC) blunted the effect of CAA. Thus, CAA seems to inhibit Na(+)/Ca(2+) exchange by interaction with cyclic adenosine monophosphate (cAMP)-PKA-signaling. A 48-hour exposure to 15 micromol/L CAA significantly reduced cell number and protein content of RPTEC by induction of necrosis. This effect of 15 micromol/L CAA could be overcome by coadministration of the intracellular Ca(2+) chelator 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM). CONCLUSION: First, CAA inhibits the Na+/Ca2+-exchanger. Second, this effect is dependent on PKA. Third, CAA induces necrotic rather than apoptotic cell death. Finally, disturbed Ca(2+) homeostasis via Na(+)/Ca(2+) exchange contributes to the nephrotoxic action of CAA in RPTEC. PMID- 16221204 TI - Curcumin blocks fibrosis in anti-Thy 1 glomerulonephritis through up-regulation of heme oxygenase 1. AB - BACKGROUND: Induction of heme oxygenase 1 (HO-1) has been shown to be beneficial in a variety of pathologic settings. Curcumin, a polyphenolic compound, has antifibrotic effects in lung models of fibrosis, and is known to induce HO-1 in renal tubular cells. In this study, we determined whether curcumin has antifibrotic properties in glomerular fibrosis and if these effects are mediated by induction of HO-1. METHODS: Curcumin effects on HO-1 expression in cultured mesangial cells and in glomeruli in vivo were analyzed by Northern and Western blotting. The dose-dependent effect of curcumin on glomerular fibrosis was tested in the anti-Thy 1 glomerulonephritis model. Curcumin was applied at doses of 10 to 200 mg/kg body weight by intraperitoneal injection from days 3 to 5 after induction of disease. On day 6, glomeruli were harvested and markers of fibrosis [plasminogen activator inhibitor-1 (PAI-1), transforming growth factor-beta (TGF beta), fibronectin, periodic acid-Schiff (PAS) staining] were analyzed. The effect of HO-1 inhibition was tested in a second experiment were nephritic rats were treated with curcumin (100 mg/kg body weight) or the combination of curcumin and the HO-1 inhibitor zinc protoporphyrin (100 microg/kg). RESULTS: Curcumin potently induced mesangial cell HO-1 expression in vitro and up-regulated glomerular HO-1 expression in nephritic animals in vivo. Curcumin treatment led to a significant, dose-dependent reduction of markers of fibrosis and proteinuria, with maximal inhibition at doses of 50 to 100 mg/kg. Beneficial effects of curcumin on markers of fibrosis and proteinuria were lost after HO-1 inhibition. CONCLUSION: Curcumin has antifibrotic effects in glomerular disease, which are mediated through an induction of HO-1. PMID- 16221205 TI - Inflammation and caspase activation in long-term renal ischemia/reperfusion injury and immunosuppression in rats. AB - BACKGROUND: We have previously shown the long-term influence of renal ischemia/reperfusion (I/R) injury and immunosuppression on fibrotic genes and apoptosis in a rat model. For the first time, we have now investigated the effects of I/R and immunosuppression on inflammation and caspase activation. METHODS: I/R injury was induced in the right kidney and the left was removed. Cyclosporin (CsA) (10 mg/kg), tacrolimus (0.2 mg/kg), rapamycin (1 mg/kg), or mycophenolate mofetil (MMF) (10 mg/kg) was then administered for 16 weeks. The effects of I/R and immunosuppressants on interstitial inflammation, interleukin (IL)-1beta expression, caspase-1 and caspase-3 activation, tubulointerstitial damage, and fibrosis were evaluated. RESULTS: ED-1+ (a specific rat monocyte/macrophage marker) cells were mainly localized in the tubulointerstitium and periglomerular areas and increased in I/R group compared to controls (P < 0.01). This was further increased by CsA, but decreased by tacrolimus, rapamycin, or MMF (P < 0.05). The 17 kD active IL-1beta remained unchanged, but 35 kD IL 1beta precursor was decreased by rapamycin in comparison with I/R group (P < 0.05). The 45 kD or 20 kD caspase-1 was increased by I/R or CsA, respectively, and decreased by rapamycin (P < 0.05). The 24 kD caspase-3, which proved to be an active caspase-3 subunit, was increased in I/R and CsA groups and deceased by tacrolimus, rapamycin, or MMF (P < 0.05), but not 32 kD precursor or 17 kD active caspase-3. The activity data of caspase-1 and caspase-3 exhibited the same trend as Western blotting data. The staining of active caspase-3 was scattered in kidneys, mainly in tubular and interstitial areas, which was consistent with that of ED-1+ cells. There was a strong positive correlation between interstitial inflammation and 24 kD caspase-3 expression or caspase-3 activity (r = 0.814 or 0.484), all of which were also closely related with urinary protein (r = 0.537, 0.529, or 0.517), serum creatinine (r = 0.463, 0.573, or 0.539), tubulointerstitial damage (r = 0.794, 0.618, or 0.712) and fibrosis (r = 0.651, 0.567, or 0.469), all P < 0.01. CONCLUSION: This study shows that the mechanisms of long-term I/R injury and immunosuppressants treatment include interstitial inflammation and caspase activation, most clearly demonstrated by the 24 kD active caspase-3. PMID- 16221206 TI - Activation of mesangial cells by platelets in systemic lupus erythematosus via a CD154-dependent induction of CD40. AB - BACKGROUND: Platelets are potential contributors to glomerular injury via the release of chemotactic and/or mitogenic mediators upon activation or through direct CD154/CD40-dependent interaction with cell components of the glomerulus. We examined whether platelets could activate mesangial cells and the potential role of the platelet-associated CD154. METHODS: Thrombin-activated platelets from systemic lupus erythematosus (SLE) patients or from disease or healthy controls were grown with human mesangial cells in the presence or not of a neutralizing anti-CD154 antibody either in contact or in a noncontact setting, the platelets and mesangial cells being separated by a pore size semipermeable membrane. The induction of mesangial cell surface antigens was assayed by flow cytometry. The quantification of mesangial cell proliferation was performed by 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and the production of transforming growth factor-beta1 (TGF-beta1), platelet-derived growth factor (PDGF) and soluble CD40 were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Activated platelets from patients with SLE could induce an up-regulation of the expression of CD40 on mesangial cells with a concomitant release of soluble CD40. This induction required a direct contact between platelets and mesangial cells and was dependent upon the platelet associated CD154. Pathologic consequences of the up-regulation of CD40 were a CD40-dependent stimulation of the proliferation of mesangial cells and a CD40 dependent increased production of TGF-beta1 by these cells. CONCLUSION: Platelets from patients with SLE can activate mesangial cells through CD40/CD154 interactions, leading to an induction of proliferation of the mesangial cells and an enhanced production of TGF-beta1, a profibrotic cytokine. PMID- 16221207 TI - Renal macrophage activation and Th2 polarization precedes the development of nephrotic syndrome in Buffalo/Mna rats. AB - BACKGROUND: At 8 weeks, Buffalo/Mna rats spontaneously develop a nephrotic syndrome associated with focal segmental glomerulosclerosis (FSGS). We have previously demonstrated that this glomerulopathy recurs after renal transplantation, thus supporting the relevance of this rat model to human idiopathic nephrotic syndrome [1]. In this study, we describe renal immune abnormalities which appear in parallel to the initiation and progression of the spontaneous Buffalo/Mna nephropathy. METHODS: Buffalo/Mna rat kidney samples were harvested before (4 weeks) and after the occurrence of proteinuria (at 10, 18, and 24 weeks, and at 12, 15, 18, and 24 months). Renal immune cell populations [total lymphocytes, macrophages, T, B, and natural killer (NK) cells] and the expression kinetics of various related cytokine [transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma), interleukin (IL)-1, IL-2, IL-4, IL-6, IL-10, IL-12, and IL-13], chemokine [regulated upon activation, normal T cell expressed and secreted (RANTES) and monocyte chemoattractant protein-l (MCP-1)] and T-cell receptor beta (TCR beta) chain transcripts were studied serially during the course of the disease. RESULTS: In the Buffalo/Mna kidneys, in parallel to the proteinuria, the focal and segmental glomerular lesions began to develop at 10 weeks (affecting 2.4 +/- 0.8% of glomeruli), increased in number, then in intensity (10.4 +/- 0.8% at 24 weeks, 14.6 +/- 2.3% at 12 months, and 28.9 +/- 7.4% at 18 months). Before the onset of the disease, at a nonproteinuric stage, the transcript expression analysis revealed a strong production of some macrophage-associated cytokines, particularly TNF-alpha (350-fold higher than control levels), which was corroborated by monocyte infiltration. A minor T-cell infiltrate (associated with an increase in Cbeta TCR transcripts), with a predominantly Th2 profile and the down-regulation of Th1 cytokines was also observed. These abnormal macrophage and T-cell patterns remained stable after the onset of the disease. No changes in chemokine and TGF-beta transcripts were observed during the initial stages of the disease. CONCLUSION: Our data suggest that the Buffalo/Mna rat disease may be the result of an immunologic disorder, involving macrophages and Th2 lymphocytes. We hypothesize that this modified environment could result in the production of a factor deleterious to the glomeruli. Thus, this rat strain could provide a new model for the study of human nephrotic syndrome. PMID- 16221208 TI - Renal tubular epithelial cells modulate T-cell responses via ICOS-L and B7-H1. AB - BACKGROUND: Renal tubular epithelial cells (TECs) play an active role in renal inflammation. Previous studies have demonstrated the capacity of TECs to modulate T-cell responses both positively and negatively. Recently, new costimulatory molecules [inducible T cell costimulator-L (ICOS-L) and B7-H1] have been described, which appear to be involved in peripheral T-cell activation. METHODS: We characterized expression and regulation of costimulatory molecules on primary human TECs and the TEC line human kidney-2 (HK-2) with reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry. Immunohistochemistry was performed on human kidney biopsies. The capacity of TECs to modulate T-cell activation was studied in TEC/T-cell cultures. RESULTS: We demonstrate that TECs express ICOS-L and B7-H1 in vitro and in vivo. Stimulation with interferon-gamma (IFN-gamma) resulted in increased expression of B7-H1, whereas ICOS-L expression was marginally increased upon stimulation with CD40L, with no effect of interleukin (IL-1), IL-17, or tumor necrosis factor-alpha (TNF-alpha). Furthermore, we show that TECs are able to costimulate T cells that have received signal-1 using alphaCD3 antibodies, inducing strong IL-10 production, which was partially mediated by ICOS-L. In contrast, B7-H1 appeared to be involved in inhibition of proliferation and cytokine synthesis. In addition, TECs were able to alter the cytokine profile of fully activated T cells, which were incubated with alphaCD3 and alphaCD28 antibodies, resulting in low IFN-gamma and high IL-10 production. This activity appeared to be independent of ICOS-L and B7-H1. CONCLUSION: Interaction of tubular epithelial cells and kidney infiltrating T cells via ICOS-L and B7-H1 may change the balance of positive and negative signals to the T cells, leading to IL-10 production and limitation of local immune responses. PMID- 16221209 TI - Expression of C-reactive protein by renal cell carcinomas and unaffected surrounding renal tissue. AB - BACKGROUND: Elevation of plasma C-reactive protein (CRP) is common in patients with renal cell carcinoma (RCC). Renal tubular epithelial cells are capable of synthesizing CRP. Although production of interleukin (IL)-6 has been described in RCC, CRP expression by carcinoma cells has yet not been investigated. METHODS: In the present study we analyzed CRP plasma levels as well as intratumoral CRP and IL-6 expression of RCC from 40 patients who underwent radical nephrectomy by means of quantitative real-time polymerase chain reaction (PCR) and immunohistochemistry. For each tumor, specimens were obtained from tumor center, tumor margin, and unaffected surrounding renal tissue. RESULTS: Preoperative plasma CRP levels correlated significantly with tumor stage (P = 0.05) and grade (P < 0.01). CRP mRNA expression was detected in 26 of 33 (79%), 30 of 36 (83%), and 32 of 36 (89%) samples from tumor center, tumor margin, and unaffected surrounding tissue, respectively. However, levels of CRP mRNA were significantly higher in tumor tissue compared to adjacent renal tissue (P < 0.01). Clear cell carcinoma exhibited significantly higher CRP mRNA levels than papillary carcinoma (P < 0.05). CRP plasma levels correlated significantly with quantitative levels of CRP mRNA within tumors (P < 0.0001). Immunohistochemically, strong CRP production was observed both in tumor cells and in tubular epithelial cells in unaffected tissue, respectively. All kidneys expressed IL-6 mRNA in the tumor and/or the unaffected tissue, but levels of intratumoral IL-6 mRNA showed no significant correlation with CRP plasma levels or local CRP transcription. CONCLUSION: In patients with RCC, a tumor-derived origin of some plasma CRP is likely. Activity of the IL-6/CRP network in RCC contributes to the accumulating evidence of the acute-phase reaction as a local inflammatory process. PMID- 16221210 TI - Soluble fibronectin induces chemokine gene expression in renal tubular epithelial cells. AB - BACKGROUND: Increasing proteinuria in kidney disease is associated with an increased risk of renal failure. Urinary proteins such as albumin induce inflammatory signaling and gene expression in tubular epithelial cells (TECs). Fibronectin is an extracellular matrix protein that can exist in soluble form and is excreted in the urine of patients with glomerular disease. METHODS: To explore the impact of soluble fibronectin on tubular epithelium, murine TECs were stimulated with soluble fibronectin and chemokine mRNA was determined by RNase protection assay. RESULTS: Fibronectin induced the expression of inflammatory chemokine genes, including monocyte chemoattractant protein-1 (MCP-1) (CCL2) and macrophage inflammatory protein-2 (MIP-2) within 2 hours in a dose-dependent manner. Phosphorylation of Src family tyrosine kinases was also increased in TECs following exposure to fibronectin. Src tyrosine kinases were involved in the fibronectin activation of MCP-1 since the Src inhibitors SU6656 and PP2 effectively reduced the induction of this chemokine. Fibronectin also induced the phosphorylation of extracellular signal-regulated protein kinase (ERK1/2) within minutes in TECs. The ERK kinase (MEK1/2) inhibitor U0126 inhibited the fibronectin induction of MCP-1 mRNA suggesting that ERK1/2 was also involved in this inflammatory pathway. Furthermore, fibronectin also induced phosphorylation of IkappaBalpha within 20 minutes in TECs. The nuclear factor-kappaB (NF-kappaB) inhibitors N-acetyl-L-cysteine (NAC) and pyrrolidinecarbodithioic acid (PDTC) effectively blocked fibronectin induction of MCP-1 mRNA. CONCLUSION: Soluble fibronectin activates MCP-1 gene expression in TECs via Src tyrosine kinases, ERK1/2 and NF-kappaB. These data provide further support to the concept that proteinuria per se contributes to the tubulointerstitial injury observed in glomerular disease. PMID- 16221211 TI - Regulation of collagen type IV genes is organ-specific: evidence from a canine model of Alport syndrome. AB - BACKGROUND: Despite advances in knowledge about collagen type IV at the protein level, little is known about expression of its six alpha chains. X-linked Alport syndrome provides a system to study collagen type IV gene expression within a setting of disturbed protein synthesis. Mutations in the alpha5 chain result in loss of the alpha3/alpha4/alpha5 and alpha1/alpha2/alpha5/alpha6 networks from the kidney, with progressive renal disease. METHODS: We used a canine model of Alport syndrome to measure expression of the six type IV collagen chains from 11 days to 7(1/2) months of age. We determined to what extent message levels in kidney change over time, and what correlation exists with clinical and pathologic changes in glomeruli, and the primary mutation. The latter was evaluated by examining testis, an organ normally containing the same collagen type IV networks but uninvolved by disease. RESULTS: The alpha1 to alpha6 mRNAs were expressed at all time points in normal canine kidney. By comparison to normal, in Alport dog kidney, the alpha1 and alpha2 mRNAs were up-regulated after 2 months of age, alpha3 and alpha4 mRNAs were down-regulated by 2 months of age, and the alpha5 mRNA was almost undetectable at any time. In testis, all mRNAs were expressed at comparable levels in normal and affected dogs other than the alpha5 chain, which was not expressed in affected testis. CONCLUSION: Normal expression of collagen type IV is under control mechanisms specific to each organ and to individual chains. The altered expression in canine Alport syndrome is not the direct result of the mutation, since these changes do not occur in all organs nor are they present from birth. Instead, collagen type IV expression is influenced by disease, with down-regulation of alpha3 and alpha4 chains temporally related to the onset of proteinuria, and up-regulation of alpha1 and alpha2 chains to glomerulosclerosis. This dysregulation of the alpha3 and alpha4 chains is unique to this Alport model, and suggests an unidentified mechanism linking pathology with down-regulation of expression of these two chains. PMID- 16221212 TI - Early treatment with cGMP phosphodiesterase inhibitor ameliorates progression of renal damage. AB - BACKGROUND: Chronic renal disease is associated with oxidative stress and reduced nitric oxide availability which, in turn, promotes hypertension and further progression of renal damage. Most actions of nitric oxide are mediated by cyclic 3',5' guanosine monophosphate (cGMP) which is rapidly degraded by phosphodiesterases (PDE). Therefore, we investigated if inhibition of PDE-5 would retard the progression of chronic renal failure. METHODS: We studied rats with 5/6 nephrectomy treated with sildenafil (2.5 mg/kg(-1)/day(-1)) in two experimental protocols. In the first protocol, we started sildenafil therapy immediately after renal ablation and continued treatment for 8 weeks. Control groups consisted of rats with renal ablation treated with drug-free vehicle and sham-operated rats with and without sildenafil treatment. RESULTS: In these studies, sildenafil treatment prevented hypertension and deterioration of renal function, reduced histologic damage, inflammation and apoptosis, delayed the onset of proteinuria, and preserved renal capillary integrity. In the second protocol we compared sildenafil with losartan (7.5 mg/kg(-1)/day(-1)) and the combination of both drugs in established renal disease, starting these drugs 4 weeks after 5/6 nephrectomy. Delayed sildenafil treatment failed to improve proteinuria and glomerulosclerosis but ameliorated hypertension and azotemia. CONCLUSION: These observations suggest that currently available PDE-5 inhibitors have potential clinical value in the treatment of chronic renal disease. PMID- 16221213 TI - Up-regulation of glomerular COX-2 by angiotensin II: role of reactive oxygen species. AB - BACKGROUND: Prostaglandins such as prostaglandin E(2) (PGE(2)) and prostaglandin I(2) (PGI(2)) counteract the angiotensin II (Ang II)-induced vasoconstriction in the glomerular microcirculation. We have shown that Ang II promotes mesangial cell hypertrophy via reactive oxygen species (ROS), which originate from nicotinamide adenine dinucleotide phosphate and its reduced form (NADH/NADPH) oxidase. It has been reported that conditions associated with activation of the renin-angiotensin system result in increased glomerular cyclooxygenase-2 (COX-2) expression and activity. METHODS: We designed studies to determine (1) whether Ang II induces COX-2 in the glomerulus in vivo in the glomerulus as well as in vitro in mesangial cells, (2) whether ROS originated from Ang II are involved, and (3) whether COX-2-derived prostaglandins modulate the growth promoting effects of Ang II in mesangial cells. Rats were infused with Ang II (0.7 mg/kg/day) for 5 days and glomerular COX-2 expression and activity assessed in isolated glomeruli. RESULTS: Ang II increased glomerular PGE(2) production (100%) accompanied by a concomitant increase in glomerular COX-2 expression at the mRNA (1.7-fold) and protein level (sixfold). In mesangial cells, Ang II significantly increased mesangial cell PGE(2) (200%) and PGI(2) (100%) production as well as COX-2 mRNA that was prevented by the angiotensin type 1 (AT1) receptor blocker irbesartan and the COX-2 inhibitor NS-398. The NADPH oxidase inhibitor diphenyleneiodonium (DPI), the ROS scavenger tiron as well as catalase, inhibited Ang II-induced PGE(2) production suggesting that Ang II-induced ROS mediate COX-2 up-regulation. Strikingly, COX-2 inhibition as well as blockade of the type 1 PGE(2) receptor (EP1) prevented Ang II-induced mesangial cell hypertrophy suggesting that COX-2-derived prostaglandins, and specifically PGE(2), importantly contribute to the growth promoting effects of Ang II. CONCLUSION: These studies suggest that blockade of specific PGE(2) receptors may be a novel strategy to modulate the pathologic effects of COX-2-derived prostaglandins without simultaneously affecting protective vasodilatory mechanisms. PMID- 16221214 TI - Loss of nitric oxide and endothelial-derived hyperpolarizing factor-mediated responses in aging. AB - BACKGROUND: Aging has considerable structural and functional effects on the vascular system of the kidney. One such effect is an alteration in vascular tone which potentially will initiate renal damage. Vascular tone is determined by the balance between vasoconstrictors and vasodilators. Therefore, we hypothesized that aging attenuates vasodilatory responses in the kidney. These changes may be mediated by a loss of nitric oxide and endothelial-derived hyperpolarizing factor (EDHF). METHODS: The systemic and renal responses of nitric oxide and EDHF were investigated in aging (18 months old) and young (3 months old) Sprague-Dawley rats. RESULTS: We demonstrated a general loss of vasodilatory responses in the aging kidney. In addition, nitric oxide levels were reduced in the serum and kidney cortex of aging versus young animals, although this was not accompanied with a loss of endothelial nitric oxide synthase (eNOS) protein in the kidney cortex. Aging animals also exhibited a loss in EDHF-mediated vasodilation following stimulation with either acetylcholine or bradykinin in the isolated perfused kidney. CONCLUSION: These findings indicate that not only a defect in the nitric oxide pathway, but also a loss of EDHF-mediated responses may be responsible for impaired vasodilation in the aging kidney. This may result in enhanced vasoconstrictive responses in aging which potentially will cause renal damage and ultimately a loss in glomerular filtration rate (GFR). PMID- 16221215 TI - Genetic variance of SGK-1 is associated with blood pressure, blood pressure change over time and strength of the insulin-diastolic blood pressure relationship. AB - BACKGROUND: Insulin stimulation of the serum- and glucocorticoid-regulated kinase 1 (SGK-1) prolongs the half-life of the epithelial sodium channel, a protein which is essential for blood pressure regulation. The aim of this study was to investigate if variation in the SGK-1 gene is associated with increased blood pressure and strength of the insulin-blood pressure relationship. METHODS: A promoter C/T, an intron 6 C/T and an exon 8 C/T polymorphism in the SGK-1 gene were genotyped in 4830 subjects from the Malmo Diet and Cancer (MDC) material of whom 4001 were free from antihypertensive medication. Of these, 2171 subjects had also been investigated 11.2 +/- 4.4 years earlier in the Malmo Preventive Project (MPP). RESULTS: In untreated MDC subjects, intron 6 CC genotype carriers had higher diastolic blood pressure than carriers of the T allele (P = 0.02) and exon 8 C allele carriers had higher systolic blood pressure than TT genotype carriers (P = 0.05). Subjects simultaneously carrying the intron 6 CC genotype and the exon 8 CC or CT genotype (SGK-1 risk) had higher systolic blood pressure (P = 0.03) and higher diastolic blood pressure (P = 0.009) than noncarriers. From MPP to MDC, the percent change in blood pressure per year was higher for systolic blood pressure (P = 0.002) and diastolic blood pressure (P = 0.001) in SGK-1 risk carriers than noncarriers. The correlation between fasting plasma insulin concentration and diastolic blood pressure was stronger in SGK-1 risk carriers than in non-carriers (P = 0.04). CONCLUSION: Our data suggest that SGK-1 risk carriers are at increased risk of hypertension and are more sensitive to the blood pressure elevating effects associated with hyperinsulinemia. PMID- 16221216 TI - Effects of nutritional status on the L-arginine-nitric oxide pathway in platelets from hemodialysis patients. AB - BACKGROUND: Malnutrition is a common feature in chronic renal failure and adversely affects patient morbidity and mortality. We here investigate the effects of nutritional status on the L-arginine-nitric oxide signaling pathway and platelet function in chronic renal failure patients on regular hemodialysis. METHODS: Platelet aggregation was correlated with plasma amino acid profiles, L arginine transport, and nitric oxide synthase (NOS) activity determined by conversion of L-[(3)H]-arginine to L-[(3)H]-citrulline and accumulation of intracellular cyclic guanosine monophospate (cGMP) in platelets from malnourished and well-nourished chronic renal failure patients on regular hemodialysis (N = 78). RESULTS: Transport of L-arginine (pmol/10(9)cells/min) via y(+) L system was increased in well-nourished (104 +/- 15) compared to controls (57 +/- 11) or malnourished chronic renal failure patients (55 +/- 13). Basal NOS activity (pmol/10(8)cells) was enhanced in well-nourished chronic renal failure patients (0.51 +/- 0.01) compared to controls (0.18 +/- 0.01) or malnourished chronic renal failure patients (0.08 +/- 0.03). In addition, basal cGMP levels are elevated in platelets from well-nourished chronic renal failure compared to malnourished uremic patients. Platelet aggregation induced by collagen is impaired in well-nourished chronic renal failure patients compared to malnourished patients and controls. Plasma L-arginine levels are reduced in chronic renal failure patients and even lower in malnourished patients. CONCLUSION: Our findings provide the first evidence that L-arginine transport via the high affinity system y(+) L and nitric oxide synthesis are only stimulated in platelets from well-nourished chronic renal failure patients, leading to impaired platelet aggregation. The absence of this adaptive response in the l-arginine nitric oxide pathway in platelets from malnourished chronic renal failure patients may account for the enhanced occurrence of thrombotic events in these patients. PMID- 16221217 TI - Kidney immune cell infiltration and oxidative stress contribute to prenatally programmed hypertension. AB - BACKGROUND: Prenatal environment has been shown to modify adult blood pressure profile, but the underlying mechanisms are not well understood. The role of renal immune cell infiltration, oxidative stress, and nitric oxide bioavailability in the pathogenesis was investigated. METHODS: Adult hypertension in rat offspring was induced by maternal low protein diet. Oxidative stress was determined by quantitative immunoblotting for nitrotyrosine, and T-cell and macrophage content by immunostaining, in offspring kidneys before and after the onset of hypertension. Nitric oxide metabolites (NOx) were measured in 24-hour urines. A group of offspring was treated with the immunosuppressive drug mycophenolate mofetil (MMF) to reduce inflammation, or with the superoxide dismutase mimetic Tempol to reduce oxidative stress, for a 3-week period before the onset of hypertension. RESULTS: During the prehypertensive stage, at 4 weeks of age, the low protein diet pups exhibited an increase in kidney nitrotyrosine content and in number of immune cells, both of which persisted in untreated animals after hypertension was established, at 8 weeks of age. Urine NOx was increased at 4 weeks and unchanged at 8 weeks of age. Both MMF and Tempol treatment prevented the immune cell infiltration, the increase in kidney nitrotyrosine abundance, and the development of hypertension. The effect on blood pressure persisted throughout the 4- to 10-week observation period after discontinuation of the treatments. CONCLUSION: Renal oxidative stress and infiltrating immune cells may play a pathogenetic role in prenatally programmed hypertension. Nitric oxide bioavailability does not appear impaired. PMID- 16221218 TI - Effects of renin-angiotensin system blockade on renal angiotensin-(1-7) forming enzymes and receptors. AB - BACKGROUND: Angiotensin-converting enzyme (ACE)2, a homologue of ACE, which is insensitive to ACE inhibitors and forms angiotensin-(1-7) [Ang-(1-7)] from angiotensin II (Ang II) with high efficiency was investigated in response to chronic blockade with lisinopril, losartan, and both drugs combined. METHODS: Thirty-six adult Lewis rats were assigned to receive these medications in their drinking water for 2 weeks while their arterial pressure, water intake, and urine volume were recorded throughout the study. Measures of renal excretory variables included assessing excretion rates of angiotensin I (Ang I), Ang II and Ang-(1-7) while blood collected at the completion of the study was used for measures of plasma angiotensin concentrations. Samples from renal cortex were assayed for renin, angiotensinogen (Aogen), neprilysin, angiotensin types 1 and 2 (AT(1) and AT(2)) and mas receptor mRNAs by semiquantitative reverse transcriptase (RT) real time polymerase chain reaction (PCR). ACE2 activity was determined as the rate of Ang II conversion into Ang-(1-7). RESULTS: Comparable blood pressure reductions were obtained in rats medicated with either lisinopril or losartan, whereas both drugs produced a greater decrease in arterial pressure. Polyuria was recorded in all three forms of treatment associated with reduced osmolality but no changes in creatinine excretion. Lisinopril augmented plasma levels and urinary excretion rates of Ang I and Ang-(1-7), while plasma Ang II was reduced with no effect on urinary Ang II. Losartan produced similar changes in plasma and urinary Ang-(1-7) but increased plasma Ang II without changing urinary Ang II excretion. Combination therapy mimicked the effects obtained with lisinopril on plasma and urinary Ang I and Ang-(1-7) levels. Renal cortex Aogen mRNA increased in rats medicated with either lisinopril or the combination, whereas all three treatments produced a robust increase in renal renin mRNA. In contrast, ACE, ACE2, neprilysin, AT(1), and mas receptor mRNAs remained unchanged with all three treatments. Renal cortex ACE2 activity was significantly augmented in rats medicated with lisinopril or losartan but not changed in those given the combination. CONCLUSION: Our data revealed a role for ACE2 in Ang-(1-7) formation from Ang II in the kidney of normotensive rats as primarily reflected by the increased ACE2 activity measured in renal membranes from the kidney of rats given either lisinopril or losartan. The data further indicate that increased levels of Ang-(1-7) in the urine of animals after ACE inhibition or AT(1) receptor blockade reflect an intrarenal formation of the heptapeptide. PMID- 16221220 TI - N-acetylcysteine attenuates the progression of chronic renal failure. AB - BACKGROUND: Lipid peroxidation impairs renal function. Aldosterone contributes to renal injury in the remnant kidney model. This study aimed to determine the effects of the antioxidant N-acetylcysteine (NAC) on renal function and aldosterone levels in chronic renal failure. METHODS: Adult male Wistar rats were submitted to 5/6 nephrectomy or laparotomy (sham-operated) and received NAC (600 mg/L in drinking water, initiated on postoperative day 7 or 60), spironolactone (1.5 g/kg of diet initiated on postoperative day 7), the NAC-spironolactone combination or no treatment. Clearance studies were performed on postoperative days 21, 60, and 120. RESULTS: Mean daily NAC and spironolactone ingestion was comparable among the treated groups. Mean weight gain was higher in NAC-treated rats than in untreated rats. A significant decrease in urinary thiobarbituric acid reactive substances (TBARS) concentrations, a lipid peroxidation marker, was observed in NAC-treated rats. By day 120, glomerular filtration rate (GFR), which dropped dramatically in untreated rats, was stable (albeit below normal) in NAC treated rats, which also presented lower proteinuria, glomerulosclerosis index, and blood pressure, together with attenuated cardiac and adrenal hypertrophy. These beneficial effects, observed even when NAC was initiated on postnephrectomy day 60, were accompanied by a significant reduction in plasma aldosterone and urinary potassium sodium [corrected] ratio. The NAC-spironolactone combination lowered blood pressure and improved GFR protection. CONCLUSION: The NAC spironolactone combination improves renal function more than does NAC alone. In the remnant kidney model, early or late NAC administration has a protective effect attributable to decreased plasma aldosterone and lower levels of lipid peroxidation. PMID- 16221219 TI - Effects of specific COX-2-inhibition on renin release and renal and systemic prostanoid synthesis in healthy volunteers. AB - BACKGROUND: The renin-angiotensin system plays a critical role in cardiovascular function, but little is known about the effects of specific cyclooxygenase 2 (COX 2) inhibition on this system in healthy humans under physiologic conditions. METHODS: Twenty-one healthy female volunteers received, in a randomized, double blind, crossover study, celecoxib 200 mg twice a day, indomethacin 50 mg three times a day, or placebo for 4 days and a single dose, each, on day 5. On day 5 of each treatment, the following parameters were assessed with subjects in an upright position before and after administration of 20 mg furosemide intravenously: plasma renin activity (PRA), plasma aldosterone, serum and urine electrolytes, and creatinine. Index metabolites of prostanoids were analyzed by gas chromatography-tandem mass spectrometry in 24-hour urine on day 4 and in 2 hour urines before and after furosemide administration. RESULTS: Baseline and furosemide-stimulated PRA were reduced to a similar degree by celecoxib and indomethacin. Plasma aldosterone and urinary excretion of potassium showed changes consistent with the alteration of PRA. Urinary excretion rates of prostaglandin E(2), (PGE(2)), 7alpha-hydroxy-5, 11-diketotetranor-prosta-1,16 dioic acid (PGE-M), and 2,3-dinor-thromboxane B(2) (TxB(2)) were not reduced by celecoxib, whereas indomethacin led to a decrease of 40%, 45%, and 80%, respectively. Both active treatments inhibited urinary excretion of 2,3-dinor-6 keto-PGF(1alpha) and 6-keto-PGF(1alpha) by 60% and 40%, respectively. CONCLUSION: Renin-release in healthy humans with normal salt intake is COX-2 dependent. While COX-1 is critical for renal and systemic PGE(2) production, renal prostacyclin synthesis is apparently COX-2 dependent. Finally, the previously demonstrated shift of the thromboxane-prostacyclin balance toward prothrombotic thromboxane by specific COX-2 inhibition is confirmed. PMID- 16221221 TI - Autosomal-dominant polycystic kidney disease in infancy and childhood: progression and outcome. AB - BACKGROUND: The natural history of autosomal-dominant polycystic kidney disease (ADPKD) has not been well described in children and infants. METHODS: The present study analyzed the characteristics of 46 ADPKD children diagnosed before 18 months of life (VEO) and 153 children diagnosed between 18 months of age and 18 years of age (non-VEO). RESULTS: VEO children had more cysts and larger renal volumes than non-VEO children when adjusted for age. In both VEO and non-VEO children, the presence of signs or symptoms at the time of diagnosis as well as the presence of hematuria or proteinuria at the study visit were associated with larger renal volumes. Children diagnosed early (VEO) or diagnosed due to signs or symptoms were also more likely to have high blood pressure. Two VEO children and no non-VEO children reached end-stage renal disease during follow-up. CONCLUSION: In contrast to many published case reports suggesting the occurrence of early end stage renal disease in VEO children, the results of the present study were much more optimistic. Over 90% of the VEO children maintained preserved renal function well into childhood. PMID- 16221222 TI - Characteristics of Chinese patients with Wegener's granulomatosis with anti myeloperoxidase autoantibodies. AB - BACKGROUND: Cytoplasmic antineutrophil cytoplasmic autoantibodies (cANCA)/proteinase-3(PR3)-ANCA was considered the serologic diagnostic marker for Wegener's granulomatosis (WG). However, Chinese patients with MPO-ANCA positive WG were frequently diagnosed. We now analyze the characteristics of patients with MPO-ANCA positive WG and investigate the difference between patients with MPO ANCA and PR3-ANCA. METHODS: Patients with WG were selected according to both Chapel Hill Consensus Conference definition and American College of Rheumatology (ACR) classification criteria in 500 Chinese patients with ANCA-associated systemic vasculitides. The clinical manifestions were compared between patients with MPO-ANCA and with PR3-ANCA. RESULTS: Eight-nine patients fulfilled the diagnostic criteria of WG: 54/89(60.7%) were MPO-ANCA positive, 34/89(38.2%) were PR3-ANCA positive. Patients with MPO-ANCA were predominantly female compared with patients with PR3-ANCA. Patients with MPO-ANCA also had multisystem involvement. However, the prevalences of arthagia, skin rash, ophthalmic and ear involvement were significantly lower in patients with MPO-ANCA than those in patients with PR3-ANCA (46.3% vs. 70.6%, P < 0.05; 20.4% vs. 44.1%, P < 0.05; 27.8% vs. 58.8%, P < 0.01; 40.7% vs. 67.6%, P < 0.05, respectively). The prevalence of elevated initial serum creatinine was significantly higher in patients with MPO-ANCA than that in patients with PR3-ANCA (81.5% vs. 61.8%, chi(2) = 4.20, P < 0.05). CONCLUSION: Patients with MPO-ANCA positive WG were not rare in Chinese. PMID- 16221223 TI - Plasma asymmetric dimethylarginine (ADMA) concentration is independently associated with carotid intima-media thickness and plasma soluble vascular cell adhesion molecule-1 (sVCAM-1) concentration in patients with mild-to-moderate renal failure. AB - BACKGROUND: Patients with renal insufficiency have an increased risk of cardiovascular disease that is not fully explained by the presence of known cardiovascular risk factors. In patients with end-stage renal disease, increased serum concentration of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase (NOS), has been linked to excess cardiovascular morbidity. We investigated, in patients with mild-to-moderate renal failure, the relationship between plasma ADMA and three surrogate markers of atherosclerosis that have been shown to have prognostic value, namely carotid intima-media thickness (IMT), plasma soluble vascular cell adhesion molecule-1 (sVCAM-1), and plasma C-reactive protein (CRP). METHODS: We used baseline data of an ongoing randomized trial in which the effects of oxidative stress-lowering treatment on vascular function and structure are studied in patients with chronic nondiabetic renal failure without clinical evidence of atherosclerosis (GFR 15 to 70 mL/min/per 1.73 m(2) according to the Cockcroft-Gault equation; ATIC study). RESULTS: Data from 93 patients were used. Creatinine clearance was inversely related to plasma ADMA concentration (standardized beta after adjustment = 0.342, P = 0.023). Plasma ADMA was strongly related to carotid IMT in univariate (beta = 0.459, P < 0.0001) and multivariate analysis (beta= 0.444, P < 0.0001). Plasma ADMA was also significantly related with plasma soluble vascular cell adhesion molecule-1 (sVCAM-1) in univariate (beta = 0.260, P = 0.010) and multivariate (beta = 0.242, P = 0.022) analysis. Plasma ADMA was not significantly related to C-reactive protein (beta = -0.134, P = 0.204). CONCLUSION: In patients with mild-to-moderate renal failure, renal function is inversely associated with plasma ADMA, which, in turn, is positively associated with carotid IMT and plasma sVCAM-1 concentration. Increased plasma ADMA may be a link between renal function and cardiovascular disease in patients with mild-to moderate renal failure. PMID- 16221224 TI - ACE I/D polymorphism is associated with mortality in a cohort study of patients starting with dialysis. AB - BACKGROUND: In dialysis patients, only a few follow-up studies have addressed the relationship between the insertion/deletion (I/D) polymorphism in the angiotensin converting enzyme (ACE) gene and mortality, but the available data are contradictory. METHODS: A cohort of 453 consecutive patients starting dialysis between January 1999 and January 2002 and participating in a Dutch multicenter prospective study was examined. Patients who died within 3 months after the start of dialysis were excluded. Patients were followed until date of death or censoring in November 2003. RESULTS: The ACE II, ID, and DD genotype frequencies were 24.3% (N = 110), 50.1% (N = 227), and 25.6% (N = 116). Besides a slightly higher number of Caucasians in the DD group, all other patient characteristics of the 3 ACE groups were similar at the start of dialysis. After adjustment for age, comorbidity, and ethnic background, patients with the ID and DD genotype showed an increased hazard ratio (HR) for all-cause mortality of 1.55 (95% CI 1.00-2.42) and 2.30 (95% CI 1.41-3.75), compared to patients with the II genotype. Slightly lower HRs were found for cardiovascular mortality. All groups of primary kidney disease showed a 2- to 3-fold increased adjusted HR for DD. CONCLUSION: The DD genotype identifies dialysis patients at an increased risk for mortality. PMID- 16221225 TI - Biventricular diastolic dysfunction in patients with autosomal-dominant polycystic kidney disease. AB - BACKGROUND: Left ventricular diastolic dysfunction has been shown in patients with autosomal-dominant polycystic kidney disease (ADPKD). However, there is no study evaluating right ventricular functions in these patients. METHODS: In the present study, diastolic functions of both ventricles in normotensive and hypertensive ADPKD patients with well-preserved renal function were investigated. Fifteen hypertensive and 16 normotensive patients with ADPKD with well-preserved renal function, 16 patients with essential hypertension, and 24 healthy subjects were included in the study. Conventional left and right ventricular echocardiographic measurements were performed in all subjects. Left and right ventricular functions were investigated both by myocardial performance index (MPI) [calculated by dividing the sum of isovolumic contraction time and isovolumic relaxation time (IVRT) by ejection time] and by tissue Doppler imaging (TDI). RESULTS: Left ventricular deceleration time and IVRT were significantly prolonged in hypertensive patients with ADPKD compared with patients with essential hypertension and even in normotensive patients with ADPKD compared with healthy subjects. Left and right MPIs were significantly higher in patients with ADPKD compared with healthy subjects, showing systolic and diastolic dysfunction. Moreover, by using TDI, the peak early diastolic mitral annular velocity (Em) to peak late diastolic mitral annular velocity (Am) ratio and the peak early diastolic tricuspid annular velocity (Et) to peak late diastolic tricuspid annular velocity (At) ratio were decreased in patients with ADPKD, suggesting biventricular diastolic dysfunction. CONCLUSION: Both hypertensive and normotensive patients with ADPKD show significant biventricular diastolic dysfunction, suggesting cardiac involvement very early in the course of ADPKD. PMID- 16221226 TI - Nephrotoxicity of low-osmolality versus iso-osmolality contrast agents: impact of N-acetylcysteine. AB - BACKGROUND: Recent data support that iodixanol, an iso-osmolality contrast agent, is less nephrotoxic than low-osmolality contrast agents when hydration is the only prophylactic strategy used. We evaluated the nephrotoxicity of iso- and low osmolality contrast agents with prophylactic administration of N-acetylcysteine (NAC) along with hydration. METHODS: Two hundred and twenty-five patients with chronic renal insufficiency (serum creatinine >1.5 mg/dL or an estimated glomerular filtration rate <60 mL/min/1.73 m(2)), referred to our institution for coronary and/or peripheral procedures, were assigned to receive low-osmolality (iobitridol group; N = 115) or iso-osmolality (iodixanol group; N = 110) contrast dye. In all cases prophylactic administration of 0.45% saline intravenously and NAC (1200 mg orally twice daily) was used. RESULTS: Baseline creatinine levels were similar in the 2 groups [iobitridol group = 1.70 (IQR: 1.54-1.98) mg/dL; iodixanol group = 1.73 (IQR: 1.56-2.00) mg/dL, P = 0.33]. The risk score for contrast nephrotoxicity was 5.0 +/- 1.6 in the iobitridol group versus 5.0 +/- 1.8 in the iodixanol group (P = 0.44). Increase of at least 0.5 mg/dL of the creatinine concentration 48 hours after the procedure occurred in 4/115 patients (3.5%) in the iobitridol group and 3/110 patients (2.7%) in the iodixanol group (P = 1.00; OR 0.78; 95% CI 0.17-3.56). Amount of contrast media administration was similar in the 2 groups (iobitridol group = 167 +/- 90 mL; iodixanol group = 164 +/- 82 mL; P = 0.61). CONCLUSION: Nephrotoxicity of iso-osmolality and low osmolality contrast agents was similar when a prophylactic strategy of hydration plus NAC was utilized. PMID- 16221227 TI - The role of osmolality in the incidence of contrast-induced nephropathy: a systematic review of angiographic contrast media in high risk patients. AB - BACKGROUND: The role of osmolality of contrast media (CM) in the pathogenesis of contrast-induced nephropathy (CIN) has been suggested by studies comparing high osmolality CM (>1500 mOsm/kg) with low-osmolality CM (550-850 mOsm/kg), and by the results of a recent comparison of a CM isotonic to plasma (iodixanol, 290 mOsm/kg) with a low-osmolality CM (iohexol, 844 mOsm/kg) in high-risk patients undergoing cardiac or peripheral angiography. METHODS: Using prospectively defined search criteria, we performed a systematic overview of prospective, randomized, controlled studies of CIN in renally impaired patients receiving intra-arterial doses of iodixanol or low-osmolality, nonionic CM, and conducted a systematic review of the data from those studies to determine whether the osmolality of CM was predictive of CIN incidence. RESULTS: Seventeen primary studies met the selection criteria, for a total of 1365 patients. Overall, the incidence of CIN was 16.8%. A multivariate logistic regression model showed that the risk of CIN is similar with the iso-osmolality iodixanol and the low osmolality iopamidol (796 mOsm/kg). The risk of CIN was significantly lower with iodixanol and iopamidol compared to iohexol. The incidence of CIN with iohexol was also significantly higher than with iopamidol, despite their similar osmolalities. CONCLUSION: These data suggest that factors other than osmolality play a significant role in the pathogenesis of CIN, at least for agents with osmolalities of 800 mOsm/kg or less. PMID- 16221228 TI - Effect of dietary modification on urinary stone risk factors. AB - BACKGROUND: This study was undertaken to ascertain the effect of dietary modification on urinary stone risks, and to determine whether the response depends on the prevailing urinary calcium. METHODS: A retrospective data analysis was conducted from our stone registry involving 951 patients with calcareous stones undergoing ambulatory evaluation, whereby 24-hour urine samples were collected during random diet and after dietary modification composed of restriction of calcium, oxalate, sodium, and meat products. Samples were analyzed for stone risk factors. Urinary calcium was also obtained after overnight fast and following a 1 g-calcium load. Changes produced by dietary modification from the random diet were evaluated in 356 patients with moderate-severe hypercalciuria (>6.88 mmol/day, group I), 243 patients with mild hypercalciuria (5.00-6.88 mmol/day, group II), and 352 with normocalciuria (<5.00 mmol/day, group III). RESULTS: Urinary calcium postcalcium load and the percentage of patients with absorptive hypercalciuria type I were highest in group I, intermediate in group II, and lowest in group III. During dietary modification, urinary calcium declined by 29% in group I, 19% in group II, and 10% in group III. Urinary oxalate did not change. Urinary saturation of calcium oxalate declined by only 12% in group I, 6% in group II, and nonsignificantly in group III, owing to various physicochemical changes in urinary biochemistry, which attenuated the effect of the decline in urinary calcium. Urinary saturation of brushite declined in all 3 groups due to the fall in urinary calcium, phosphorus, and pH. This reduction was more marked in the hypercalciuric groups than in the normocalciuric group. Urinary saturation of monosodium urate also decreased from a decline in urinary sodium and uric acid. CONCLUSION: Secondary rise in urinary oxalate occurring from calcium restriction can be avoided by concurrent dietary oxalate restriction. Dietary modification (restriction of dietary calcium, oxalate, sodium, and meat products) is more useful in reducing urinary saturation of calcium oxalate among patients with hypercalciuria than among those with normocalciuria. PMID- 16221229 TI - Identifying critically ill patients at high risk for developing acute renal failure: a pilot study. AB - BACKGROUND: Acute renal failure (ARF) occurs commonly in the intensive care unit (ICU), but predicting which patients will develop ARF is difficult. We set out to determine which risk factors would predict the development of ARF in critically ill patients who are admitted to the ICU without ARF. METHODS: From August 2002 to April 2003, we enrolled medical-surgical ICU admissions into a cohort using a sampling tool based on their risk factor (RF) profile. The risk factors we identified were separated into 3 categories: chronic major, chronic minor, and acute RFs. Combinations of these RFs were used to create a sampling tool and identify patients to enroll into our cohort. Patients with end-stage renal disease and ARF upon admission to the ICU were excluded. RESULTS: We enrolled 194 patients over a 14-month period. The mean age of the cohort was 64.6 +/- 14.7 years. The percentage of Caucasians, African Americans, and Hispanics was 40.7%, 50.5%, and 3.6%, respectively. In a univariate analysis of the entire cohort, increasing APACHE II quartile, increased A-a gradient, presence of systemic inflammatory response syndrome (SIRS), decreased levels of serum albumin, and presence of active cancer predicted ARF. In a multiple logistic regression analysis, decreased serum albumin (high levels of serum albumin were protective), increased A-a gradient, and cancer were associated with development of ARF (OR 2.17, 1.04, and 2.86, respectively). CONCLUSION: Decreased levels of serum albumin concentration, increased A-a gradient, and presence of active cancer predict which patients who are admitted to the ICU will develop ARF. PMID- 16221230 TI - Urinary ortho-tyrosine excretion in diabetes mellitus and renal failure: evidence for hydroxyl radical production. AB - BACKGROUND: Phenylalanine is converted to para- and ortho-tyrosine by hydroxyl free radical, or to para-tyrosine by the phenylalanine hydroxylase enzyme. The aim of this study was to measure para- and ortho-tyrosine in the urine and plasma of patients with chronic renal disease and/or diabetes, to obtain information on the renal handling of the different tyrosine isomers and, furthermore, to measure urinary levels of 8-epi-prostaglandin-F(2alpha), a marker of lipid peroxidation. METHODS: In our cross-sectional study we measured para-, ortho-tyrosine, and phenylalanine levels, using high performance liquid chromatography and 8-epi prostaglandin-F(2alpha) with enzyme-linked immunosorbent assay (ELISA). We compared 4 groups: (1) controls (CONTR, N = 14), (2) patients with chronic kidney disease (CKD, N = 12), (3) patients with type 2 diabetes mellitus (DIAB, N = 17), (4) patients with chronic kidney disease and type 2 diabetes (DIAB-CKD, N = 19). RESULTS: We found a decreased plasma para-tyrosine level and decreased urinary para-tyrosine excretion in CKD patients, while the fractional excretion of para tyrosine was similar in all 4 groups, approximately 1%. There was no difference in the plasma ortho-tyrosine levels between the groups. However, urinary ortho tyrosine excretion was higher in all 3 groups of patients than in the CONTR group, and higher in DIAB and in DIAB-CKD patients than in CKD patients. The fractional excretion of ortho-tyrosine was significantly higher in DIAB and in DIAB-CKD patients than in the CONTR group. The fractional excretion of ortho tyrosine exceeded 100% in the 2 diabetic groups. Urinary 8-epi-prostaglandin F(2alpha)/creatinine ratio did not correlate with urinary ortho-tyrosine excretion. CONCLUSION: The difference between para-tyrosine levels of the groups is probably due to renal impairment, while there is indirect evidence for an increased tubular secretion or production of ortho-tyrosine in the kidney in diabetic patients with or without CKD. PMID- 16221231 TI - Class IV-S versus class IV-G lupus nephritis: clinical and morphologic differences suggesting different pathogenesis. AB - BACKGROUND: A recently proposed reclassification of lupus nephritis divides class IV (diffuse proliferative) lupus nephritis into those cases with predominantly segmental proliferative lesions (class IV-S) and those with predominantly global proliferative lesions (class IV-G). This report explores the validity of this distinction and possible differences in pathogenesis between the 2 types of lesions. METHODS: Patients from a previously reported series of severe lupus nephritis, with initial biopsies (Bx1) and control biopsies (Bx2) at 6 months after induction therapy were reclassified according to the newly proposed classification. From the original series of 65 patients, 15 patients were reclassified as having class IV-S lesions and 31 patients class IV-G lesions. Clinical data at both biopsies and follow-up were available on all patients selected. RESULTS: Patients with IV-G lesions had worse proteinuria, lower serum hemoglobins, lower CH50s, and likely higher SCrs (P = .06) and lower C3s (P = .08) than class IV-S patients. Serum CH50 and C3 correlated negatively with severity of class IV-G lesions, but not at all with class IV-S lesions. Patients with class IV-G lesions had greater overall immune deposits and subendothelial deposits on IF and greater hyaline deposits on light microscopy. By contrast, class IV-S showed predominant mesangial deposits and a much higher rate of glomerular fibrinoid necroses (13.3 +/- 15.3% vs. 5.6 +/- 8.0% of viable glomeruli, P = .03). Other distinctions included the fact that membranoproliferative features were found only in class IV-G lesions, and glomerular monocyte/macrophages were much more frequent in this group than in class IV-S lesions (1.77 +/- 0.92 vs. 0.86 +/- 0.77, P = .008). Finally, class IV G frequently involved all viable glomeruli (74.2% of cases), whereas segmental proliferative lesions never did (P < .0001). Survivals from doubling of SCr at 10 years did not differ between the 2 types at Bx1: 72.5% segmental versus 60.4% global, P= .53. However, among those with persistent lesions at Bx 2 (11 IV-S and 9 IV-G), there was a dramatic difference in 10-year survivals between IV-S lesions (63.6%) and IV-G lesions (0%), P = .08. CONCLUSION: There are definite clinical and morphologic differences between class IV-S and IV-G lesions. Data suggest that class IV-G lesions behave as an immune complex disease, having positive correlations with extent of immune deposits and negative correlations with serum complement levels, the model traditionally assumed for lupus nephritis as a whole. However, in class IV-S lesions, the presence of proportionally greater glomerular fibrinoid necroses and lack of correlation with extent of immune deposits suggest that these lesions may have a different pathogenesis. PMID- 16221232 TI - Immunogenicity and safety of an adjuvanted hepatitis B vaccine in pre hemodialysis and hemodialysis patients. AB - BACKGROUND: Due to their impaired immune system, patients with renal insufficiency have a suboptimal response to hepatitis B (HB) vaccination and frequent boosters are needed to maintain protection. GlaxoSmithKline Biologicals has developed a HB vaccine containing a new adjuvant system AS04 for use in this immunocompromised patient population. METHODS: In an open, randomized clinical trial conducted in pre-hemodialysis (documented creatinine clearance < or =30 mL/min) and hemodialysis patients, over 15 years of age and naive for HB, the immunogenicity and safety of single doses of HB-AS04 (Fendrix, GlaxoSmithKline Biologicals) were compared to double doses of commercially available HB vaccine (Engerix, GlaxoSmithKline Biologicals) administered at 0, 1, 2, and 6 months, and followed-up for 36 months. RESULTS: The HB-AS04 vaccine elicited a more rapid onset of protection than the currently licensed vaccine for this particular population, with 74% versus 52% of subjects seroprotected at month 3. After the vaccination course, seroprotection rates increased to 91% versus 84% in the HB AS04 and standard vaccine groups, respectively. Differences persisted up to 36 months post-vaccination (73% vs. 52%, respectively). Antibody concentrations were higher following the HB-AS04 vaccine at all post-vaccination time points. During the follow-up, significantly fewer subjects primed with the HB-AS04 vaccine needed a booster dose as a consequence of anti-HBs loss below seroprotective levels (11/62 subjects in the HB-AS04 group vs. 22/57 subjects in the standard vaccine group, respectively, P = 0.014). The HB-AS04 was more locally reactogenic than the standard immunization regimen, with pain at the injection site occurring with 41% of HB-AS04 doses versus 19% of standard vaccine doses. The occurrence of grade 3 pain was less than 1% in both groups and all events resolved within the 4 day follow-up period. CONCLUSION: The improved immunogenicity profile and clinically acceptable reactogenicity of HB-AS04 vaccine are of key importance to provide a more rapid, enhanced, and longer seroprotection to these immunocompromised patients at risk for HB infection. PMID- 16221233 TI - Reduced bone mineral density in adults treated with high-dose corticosteroids for childhood nephrotic syndrome. AB - BACKGROUND: Children with minimal change nephrotic syndrome (MCNS) receive repeated courses of high dose oral prednisolone. No previous study has investigated the impact of this on final bone mineral density (BMD). Young adults previously reported in a large follow-up study of children with MCNS were invited to participate in a cross-sectional study. Areal BMD (aBMD) of the spine (L1-4), left femoral neck, and total left hip was measured using dual x-ray absorptiometry (DXA), and volumetric BMD (vBMD) of the distal radius was measured by pQCT. BMD results were compared with reference data provided by the manufacturers of the densitometers. METHODS: Thirty-four (24 male) of the original cohort of 62 participated in the study. The mean (SD) final height Z score of the cohort was -0.45 (0.92) (P = 0.007) and mean BMI Z score 1.62 (1.53) (P < 0.0001). RESULTS: There was a highly significant reduction in distal radial trabecular vBMD; the mean Z score was -0.95 (0.99) and T score -1.04 (1.01) (both P < 0.0001); however, there was no reduction in distal radial total vBMD, the mean Z score being 0.00 (0.95) and T score -0.08 (0.99), (P = 0.99 and 0.66, respectively). The aBMD of the lumbar spine and femoral neck also showed a reduction in T scores [-0.45 (1.27), P = 0.045 and -0.49 (0.86), P = 0.002, respectively], but not Z scores [-0.37 (1.28) and -0.15 (0.87), respectively, both P = NS]. Total hip aBMD was not different from the control population. CONCLUSION: Adult survivors of childhood MCNS have a significant reduction in forearm trabecular vBMD, placing them at increased fracture risk at this site. PMID- 16221234 TI - The conundrum of increased burden of end-stage renal disease in Asians. AB - BACKGROUND: Few cohort studies have examined the risk of end-stage renal disease (ESRD) among Asians compared with whites and blacks. METHODS: To compare the incidence of ESRD in Asians, whites, and blacks in Northern California, we examined sociodemographic and clinical data on 299,168 adults who underwent a screening health checkup at Kaiser Permanente between 1964 and 1985. Incident cases of ESRD were ascertained by matching patient identifiers with the nationally comprehensive United States Renal Data System ESRD registry. RESULTS: Overall, 1346 cases of ESRD occurred during 7,837,310 person-years of follow-up. The age-adjusted rate of ESRD (per 100,000 person-years) was 14.0 [95% confidence interval (CI) 10.5-18.5] among Asians, 7.9 (95% CI 6.5-9.5) among whites, and 43.4 (95% CI 36.6-51.4)] among blacks. Controlling for age, gender, educational attainment, diabetes, prior myocardial infarction, serum creatinine, systolic and diastolic blood pressure, proteinuria, hematuria, cigarette smoking, serum total cholesterol, and body mass index increased the risk of ESRD in Asians relative to whites from 1.69 to 2.08 (95% CI 1.61-2.67). By contrast, adjustment for the same covariates decreased the risk of ESRD in blacks relative to whites from 5.30 to 3.28 (95% CI 2.91-3.69). CONCLUSION: Factors contributing to the excess ESRD risk in Asians relative to whites extend beyond usually considered sociodemographic and comorbidity disparities. Strategies aimed at examining novel risk factors for kidney disease and efforts to increase awareness of kidney disease among Asians may reduce ESRD incidence in this high-risk group. PMID- 16221235 TI - Total body water volume for white children and adolescents and anthropometric prediction equations: the Fels Longitudinal Study. AB - BACKGROUND: There are few studies of total body water (TBW) volume in children. Such studies are needed, as are new prediction equations for the clinical management of children with renal insufficiency and those receiving dialysis. METHODS: Mixed longitudinal data were from 124 white boys and 116 white girls 8 to 20 years of age. TBW volume was measured by deuterium nuclear magnetic resonance spectroscopy, and random effects models were used to determine patterns of change over time. Sex-specific TBW prediction equations were developed using regression analysis. RESULTS: Boys had significantly greater (P < 0.05) mean TBW volumes than girls at all but 3 ages. TBW was significantly (P < 0.05) associated with age and maturation in the boys and the girls. In boys, mean TBW/WT varied from 0.55 to 0.59, while in the girls the mean declined from 0.53 to 0.49 by 16 years of age. Boys had significantly larger means for TBW/WT than girls, who had a significant, slight negative trend with age. The prediction equations were TBW = -25.87 + 0.23 (stature) + 0.37 (weight) for boys and TBW =-14.77 + 0.18 (stature) + 0.25 (weight) for girls. CONCLUSION: Means are provided for TBW in white children from 8 to 20 years of age, whose average fatness affected the percentage of TBW in body weight. These updated TBW prediction equations perform better than those available from the past. PMID- 16221236 TI - Anemia and mortality in hemodialysis patients: accounting for morbidity and treatment variables updated over time. AB - BACKGROUND: The objective of this study was to gain insight into the associations of anemia with mortality among maintenance hemodialysis (HD) patients and patient subgroups by an analysis that more comprehensively represents hemoglobin (Hb) level, morbidity, and treatment characteristics over time than was possible in prior observational studies. METHODS: A cohort study was conducted among 5517 subjects in the American arm of the Dialysis Outcomes and Practice Patterns Study Phase I. We used proportional hazard analysis to model all-cause mortality as a function of Hb level measured 1, 3, and 6 months previously. Forty-five potentially confounding patient-level characteristics were considered, including demographics, comorbidities, and time-updated levels of erythropoietin and parenteral iron dosing, medical events, and laboratory and dialysis measures. RESULTS: Compared to Hb 11 to <12 g/dL, subjects with Hb <11 g/dL had increased mortality [adjusted hazard ratios (95% confidence interval) in the 3-month-lagged model = 1.74 (1.24 to 2.43) for <9 g/dL, 1.25 (0.96 to 1.63) for 9 to <10 g/dL, and 1.22 (0.99 to 1.49) for 10 to <11 g/dL categories]. Mortality rates for subjects with Hb 12 to <13 g/dL and > or = 13 g/dL did not differ significantly from those with Hb 11 to <12 g/dL. The relationships between Hb and mortality varied modestly with changes in the time interval between Hb measurement and the time at risk for mortality, but did not vary according to ESRD vintage or health status indicators. CONCLUSION: Our findings confirm the associations of Hb levels > or =11 g/dL with longer survival among maintenance HD patients, but show no additional survival advantage for patients with Hb levels > or =12 g/dL. Further investigation of the relationships among anemia, treatment of anemia, and survival is warranted. PMID- 16221237 TI - Convective and adsorptive removal of beta2-microglobulin during predilutional and postdilutional hemofiltration. AB - BACKGROUND: Beta(2)-microglobulin (beta2-m) removal in patients with end-stage renal disease (ESRD) is maximal with convective techniques, such as hemofiltration (HF) or hemodiafiltration (HDF). Although the infusion mode of the replacement solution (predilution or postdilution) is expected to influence the efficiency of HF, experimental data in this respect are scanty. We therefore investigated the impact of the fluid reinfusion mode on the efficiency of HF in 11 ESRD patients who underwent both treatments. METHODS: The dialyzer (AK 200 ULTRA) was equipped with a 3-layer polyamide membrane (Poliflux 21 S, surface 2.1 m(2)) and blood flow was kept between 300 and 400 mL/min. beta2-m concentrations were measured in plasma water and ultrafiltrate at appropriate times during a 240 minute treatment. The following dialytic parameters were calculated: total amount of beta2-m removed (A(tot)), beta2-m removed by convection (A(con)) and by adsorption (A(ads)), percent reduction in beta2-m plasma water concentration (% Cpw(in)), total plasma water clearance (CLpw(tot)), convective plasma water clearance (CLpw(con)), adsorptive plasma water clearance (CLpw(ads)), and sieving coefficient (SC). RESULTS: CLpw(tot), CLpw(ads), and% Cpw(in) were similar in pre and postdilutional conditions, whereas CLpw(con) and SC were higher and CLpw(ads) was lower in postdilution than in predilution HF. Since a significant inverse correlation was found between A(ads) and SC, predilution probably determines greater protein fouling than postdilution. CONCLUSION: The 2 techniques appear to be equivalent in terms of total beta2-m removal, although this final result is obtained by different contributions of convective and adsorptive elimination. PMID- 16221238 TI - Skeletal muscle, cytokines, and oxidative stress in end-stage renal disease. AB - BACKGROUND: End-stage renal disease (ESRD) is a state of microinflammation, with increased activation of cytokines and augmented oxidative stress. While peripheral blood mononuclear cells are an established source of reactive oxygen species and inflammatory cytokines during hemodialysis (HD), skeletal muscle is also capable of generating these biomolecules. METHODS: Femoral arterio-venous (A V) balance of interleukin-1 (IL-1), IL-6, IL-10, tumor necrosis factor-alpha (TNF alpha), malonyldialdehyde (MDA), and carbonyl protein (CP) were measured in 17 ESRD patients and 9 healthy volunteers. ESRD patients were studied before (pre HD) and during HD. mRNA levels of cytokines, heme oxygenase-1 (HO-1), and suppressors of cytokine signaling-2 (SOCS-2) were quantitated in the skeletal muscle by real-time polymerase chain reaction (PCR). RESULTS: Arterial concentration of MDA (pmol/mL) was higher pre-HD (325.5 +/- 19.6) compared to controls (267.7 +/- 14.7), but decreased intradialysis (248.8 +/- 16.1) (P < 0.01). Dialysis clearance of MDA was 16.9 +/- 3.1 mL/min. CP concentration (nmol/mg protein) in the artery was significantly higher pre-HD (2.29 +/- 0.09) than in controls (1.92 +/- 0.05), and remained stable during HD (2.23 +/- 0.07). Plasma cytokines increased to a variable degree in the artery and vein during HD. A-V balance studies demonstrated that the MDA (17.8%) and CP (5.1%) concentrations increased significantly in the vein intradialysis. Venous concentration of IL-6 was higher than that in the artery during dialysis (16.27 +/- 2.42 vs. 11.29 +/- 2.17 pg/dL, P < 0.01). mRNA levels of IL-6 (0.028 +/- 0.02 vs. 6.69 +/- 0.21), HO-1 (0.96 +/- 0.01 vs. 5.08 +/- 1.11), and SOCS-2 (0.63 +/- 0.12 vs. 0.82 +/- 0.14) in the muscle increased during HD (P < 0.01). Immunohistochemical studies confirmed the increase in IL-6 protein in the skeletal muscle during HD. The intradialytic increase in IL-1, IL-10, and TNF alpha gene expression was not significant. CONCLUSION: Skeletal muscle may also contribute to the circulating plasma IL-6 and increased oxidative stress during HD. PMID- 16221239 TI - The impact of waiting time and comorbid conditions on the survival benefit of kidney transplantation. AB - BACKGROUND: Longer waiting times may limit the survival benefit of kidney transplantation in older patients or those with a high burden of comorbid disease. METHODS: We performed a longitudinal study of mortality among 63,783 transplant candidates who started dialysis between April 1995 and December 2000. We determined the relative risk (RR) of death and increase in life expectancy among subjects who received a first deceased donor transplant after different waiting times compared to subjects who had equivalent waiting times but remained on dialysis. RESULTS: Transplant recipients had a lower long-term RR of death and the risk reduction was greatest in recipients with longer waiting times (RR of death 12 months after transplantation for recipients with waiting times of 0, 1, 2, 3 years was 0.49, 0.43, 0.38, 0.34, P = 0.0006). The average increase in life expectancy in transplant recipients was 9.8 years and was lower in older recipients and recipients with comorbid conditions. Increased waiting times from 1 to 3 years only moderately decreased the overall survival benefit of transplantation from 7.1 to 5.6 years, and all subjects derived a survival benefit from transplantation with waiting times up to 3 years. CONCLUSION: These findings do not support limiting access to transplantation for otherwise suitable candidates on the basis of longer anticipated waiting times. PMID- 16221240 TI - Monthly access flow monitoring with increased prophylactic angioplasty did not improve fistula patency. AB - BACKGROUND: Regular access monitoring is recommended to detect and treat access stenosis in order to prevent access thrombosis and failure. METHODS: In 1999, we instituted monthly access blood flow monitoring using the ultrasound dilution technique (UDT). In a sequential observational trial, 222 patients were studied for the impact of UDT monitoring on patency of their first arteriovenous autogenous fistula. Group 1, the historic group (before 1999), had 146 arteriovenous fistulas (50.7% upper arm), followed for 259 access-years. Group 2, the UDT-monitored group, had 76 arteriovenous fistulas (60.5% upper arm), followed for 123 access-years. Decision to refer for angiography was based on clinical criteria for group 1, and clinical criteria plus results of UDT flow monitoring in group 2. RESULTS: Cumulative patency was longer (P < 0.01) and the thrombosis rate was lower (P < 0.05) in group 2. However, the improvement occurred prior to initiation of UDT flow monitoring. Comparing outcomes in group 2 patients whose fistula survived to start flow monitoring with group 1 patients whose fistula survived at least 160 days (the median time to starting UDT monitoring in group 2), there was a sevenfold increase in angioplasty procedures (0.67 vs. 0.09 per access-year) but no improvement in the thrombosis rate or cumulative fistula patency. CONCLUSION: UDT monitoring increased the rate of angioplasty procedures and thereby shortened primary unassisted patency, but did not decrease the thrombosis rate or improve cumulative fistula patency. PMID- 16221241 TI - Apparent successful mesothelial cell transplantation hampered by peritoneal activation. AB - BACKGROUND: Mesothelial cell transplantation has been suggested to improve mesothelial repair after surgery, recurrent peritonitis and peritoneal dialysis. METHODS: In this study we evaluated mesothelial cell transplantation during the resolution phase of experimentally thioglycollate-induced peritonitis in rats. To this end 4 x 10(6) DiO-labeled autologous mesothelial cells were transplanted 1 week after peritonitis induction. Peritoneal inflammation and permeability characteristics were evaluated after another week. RESULTS: Mesothelial cell transplantation after peritonitis resulted in incorporation of these cells in the parietal mesothelial lining, leading to an acute transient submesothelial thickening which was not seen in transplanted animals without prior peritonitis induction. Long-term functioning of these repopulated mesothelial cells leaded to peritoneal activation as evidenced by a approximately twofold increase in peritoneal lymphocytes (P < 0.01) and omental mast cell counts (P < 0.05), accompanied by the induction of inflammation markers monocyte chemoattractant protein-1 (MCP-1) (P < 0.01) and hyaluronan (P < 0.01) in the transplanted peritonitis group, but not in rats with peritonitis without mesothelial cell transplantation or in control rats without mesothelial cell transplantation (all four parameters P < 0.01). In addition, trapping of transplanted mesothelial cells in the milky spots of omental tissue and lymphatic stomata of the diaphragm both in control and thioglycollate rats seems to increase microvascular permeability, reflected by apparent increased diffusion rates of small solutes and proteins. CONCLUSION: Altogether, our data underscore the importance of controlling peritoneal (patho)physiology and function in mesothelial transplantation protocols. PMID- 16221242 TI - Accelerated lean body mass loss in incident chronic dialysis patients with diabetes mellitus. AB - BACKGROUND: End-stage renal disease (ESRD) patients display a higher incidence of poor nutritional status and are at high risk of hospitalization and death. Patients on renal replacement therapy (RRT) with a primary diagnosis of diabetes mellitus have the lowest survival rates along with highest hospitalization incidence. METHODS: In this study, we examined the importance of diabetes mellitus along with certain demographic and clinical variables in predicting the change in lean body mass (LBM) by dual-energy x-ray absorptiometry (DEXA), as a surrogate marker of somatic protein stores, in 142 incident ESRD patients (91 males, 52.8 +/- 1.0 years, 74.2 +/- 1.2 kg body weight) among which 34 had diabetes mellitus (19 insulin-dependent and 15 noninsulin dependent). RESULTS: Our results show that patients with diabetes mellitus had significantly accelerated loss of LBM compared to nondiabetic patients during the first year of RRT (3.4 +/- 0.6 kg vs. 1.1 +/- 0.2 kg) (P < 0.05). Multivariate linear regression analyses revealed that the presence of diabetes mellitus was the strongest predictor of LBM loss independently of several clinically-relevant variables such as age, gender, serum albumin, presence of malnutrition, presence of inflammation, and RRT modality. CONCLUSION: We conclude that the presence of diabetes mellitus is the most significant independent predictor of LBM loss in renal replacement therapy patients, providing a potential explanation as to why ESRD patients with diabetes mellitus are more prone to muscle wasting. PMID- 16221243 TI - Cefazolin plus netilmicin versus cefazolin plus ceftazidime for treating CAPD peritonitis: effect on residual renal function. AB - BACKGROUND. The International Society for Peritoneal Dialysis (ISPD) treatment guidelines for continuous ambulatory peritoneal dialysis (CAPD) peritonitis 2000 recommended the use of cefazolin plus ceftazidime as the initial empirical therapy in patients with residual renal function (RRF). However, this treatment regimen has not been compared with the conventional regimen of cefazolin plus netilmicin in prospective, randomized controlled trials. METHODS: Stable CAPD patients who developed clinical evidence of peritonitis were randomized to receive intraperitoneal (i.p.) cefazolin plus netilmicin or cefazolin plus ceftazidime once daily in the long dwell for 14 days. For patients with RRF (>1 mL/minute) before entry into the study (N= 50), RRF and 24-hour urine volume were measured at days 1, 14, and 42 after commencement of i.p. antibiotic treatment. RESULTS: One hundred and two patients were recruited into the study. The primary cure rates of i.p. cefazolin plus netilmicin and cefazolin plus ceftazidime were 66.7% and 64.7%, respectively. The overall cure rate for the 2 treatment regimens was 82.3% for both. Seven patients (14%) from each treatment group required removal of the dialysis catheters due to treatment failure. Relapse of peritonitis occurred in 2 patients (4%) in both treatment groups. Thirty-six patients with RRF at baseline achieved primary cure of their peritonitis by the assigned antibiotics. In this subgroup of patients, their RRF and daily urine volume showed significant reduction at day 14 and returned to near baseline values at day 42. The degree of reduction in RRF and urine volume did not differ significantly between the patients treated with cefazolin plus netilmicin and cefazolin plus ceftazidime. CONCLUSION: Intraperitoneal cefazolin plus netilmicin and cefazolin plus ceftazidime have similar efficacy as empirical treatment for CAPD peritonitis. In CAPD patients with RRF, significant but reversible reduction in RRF and 24-hour urine volume could occur after an episode of peritonitis, despite successful treatment by i.p. antibiotics. The effect of i.p. cefazolin plus netilmicin, or i.p. cefazolin plus ceftazidime on RRF in CAPD patients with peritonitis does not appear to be different. Our findings do not support the routine use of cefazolin and ceftazidime as the empirical treatment for CAPD peritonitis. PMID- 16221244 TI - Single-center experience of encapsulating peritoneal sclerosis in patients on peritoneal dialysis for end-stage renal failure. AB - BACKGROUND: Encapsulating peritoneal sclerosis (EPS) is a rare but serious complication in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) or automated peritoneal dialysis (APD). It is characterized by a progressive, intra-abdominal, inflammatory process resulting in sheets of fibrous tissue that cover, bind, and constrict the viscera, thereby compromising the motility and function of the bowel. Although recent therapeutic approaches have been reported with variable success, the ability to detect reliably at an early stage patients at risk for EPS would be beneficial and allow treatment standardization. The aim of this study was to evaluate the clinical features of EPS and identify possible risk factors for its development in CAPD and APD patients. METHODS: This was a review of all cases of EPS in a single center over the last 5 years. RESULTS: There were 810 CAPD and APD patients, managed in our program over this period. We identified 27 cases of EPS, giving an overall of 3.3% in this population. The mean duration of CAPD before diagnosis of EPS was 72.6 +/- 39.7 months (range 16-172). Sixteen cases required surgical treatment and were classified as severe; others were treated conservatively (mild to moderate group). Ten patients received tamoxifen treatment with apparent benefit. The overall mortality rate was 29.6%. Eight patients from the severe group and the entire moderate group survived on hemodialysis or transplantation at 48.71 and 27.63 months follow-up, respectively. Peritonitis rates were not different between the 2 groups and peritoneal history was unremarkable compared to overall peritonitis rates in the unit. Data on small solute transport were not available in all patients in this retrospective analysis. CONCLUSION: EPS is a serious, life-threatening complication of CAPD. Most cases had PD duration of more than 4 years. Careful monitoring by CT scans of the peritoneal membrane in patients beyond 5 years, and early catheter removal in patients with peritoneal thickening should be considered for long-term CAPD patients. Treatment with tamoxifen may be of benefit in these patients. PMID- 16221245 TI - Relationship between urea clearance and ionic dialysance determined using a single-step conductivity profile. AB - BACKGROUND: On-line determination of ionic dialysance (ID) has been used to measure the clearance of small solutes like urea. However, attempts to determine the in vivo relationship between ID and urea clearance have led to discordant findings. The aim of this study was to determine the relationship between the mean values of repeated instantaneous determinations of ID throughout a dialysis session ((m)ID), obtained using a single-step inlet dialysate conductivity profile, and the mean values of urea clearance corrected for access recirculation (K(eu1)), total recirculation (access plus cardiopulmonary recirculation, K(eu2)), and the entire postdialysis urea rebound (K(wb)). METHODS: Eighty-two anuric patients on chronic thrice-weekly hemodialysis were studied using an Integra machine equipped with the Diascan module for the automatic determination of ID. The mean values of repeated ID measurements made at 30-minute intervals were compared with K(eu1) (available for only 31 patients), K(eu2), and K(wb). RESULTS: The results in all 82 patients were: (m)ID = 176 +/- 23 mL/min; K(eu2) = 181 +/- 25 mL/min; K(wb) = 159 +/- 22 mL/min. The mean (m)ID/K(wb) and (m)ID/K(eu2) ratios were, respectively, 1.11 +/- 0.06 and 0.98 +/- 0.06. The results in the 31 patients for whom K(eu1) values were available were: (m)ID = 179 +/- 24 mL/min and K(eu1) = 200 +/- 27 mL/min; the mean (m)ID/K(eu1) ratio was 0.90 +/- 0.05. CONCLUSION: The mean value of repeated ID determinations obtained using a single-step conductivity profile underestimates urea clearance corrected for access recirculation, and may be considered an adequate estimate of urea clearance corrected for total recirculation. PMID- 16221246 TI - Diabetes, a cause of progressive sarcopenia in dialysis patients? PMID- 16221247 TI - Polycystic kidney disease: a new perspective from the beginning. PMID- 16221250 TI - Questions about graft surveillance. PMID- 16221249 TI - Antibodies against macrophages that overlap in specificity with fibroblasts. PMID- 16221251 TI - Hepatitis C infection and proteinuria. PMID- 16221253 TI - Renal replacement therapy (RRT) of diabetics with end-stage renal disease (ESRD): eight years' observation from single center in Poland. PMID- 16221254 TI - Risk of bone loss or fracture among renal transplant recipients: race and steroid. PMID- 16221255 TI - Vasopressin antagonists in polycystic kidney disease. PMID- 16221256 TI - Three year outcome of cardiac resynchronization therapy: a single center evaluation. AB - AIMS: To evaluate the long-term clinical outcome and device performance of cardiac resynchronization therapy in a consecutive sample of patients with moderate to severe heart failure. METHODS AND RESULTS: Between 1998 and 2000, forty consecutive patients with drug-refractory heart failure due to ischemic or idiopathic dilated cardiomyopathy were selected for cardiac resynchronization therapy (CRT). After successful implantation of the coronary sinus lead (n = 35, 88%), patients were followed every sixth month by New York Heart Association (NYHA) functional class, the 6-minute walking test (6 walk), quality of life (QoL, Minnesota), and pacemaker control. NYHA-class and 6 walk were significantly improved after 6 months and continued to improve gradually until 36 months of follow-up. The QoL improvement at 6 months was sustained over 3 years. After 3 years, the beta-blocker dose could be increased in 10/23 patients as compared to baseline. Nine patients had to be re-operated. Coronary sinus lead thresholds were stable over time. CONCLUSION: The clinical improvements by CRT are sustained over 3 years of follow-up. In the setting of a University Medical Center, CRT can be applied in clinical routine with excellent clinical outcome, acceptable implantation success, and stable device performance over time. PMID- 16221257 TI - Transvenous access to the pericardial space: an approach to epicardial lead implantation for cardiac resynchronization therapy. AB - BACKGROUND: Percutaneous access to the pericardial space (PS) may be useful for a number of therapeutic modalities including implantation of epicardial pacing leads. We have developed a catheter-based transvenous method to access the PS for implanting chronic medical devices. METHODS: In eight pigs, a transseptal Mullins sheath and Brockenbrough needle were introduced into the right atrium (RA) from the jugular vein under fluoroscopic guidance. The PS was entered through a controlled puncture of the terminal anterior superior vena cava (SVC) (n = 7) or right atrial appendage (n = 1). A guidewire was advanced through the transseptal sheath, which was then removed leaving the wire in PS. The guidewire was used to direct both passive and active fixation pacing leads into the PS. Pacing was attempted and lead position was confirmed by cine fluoroscopy. Animals were sacrificed acutely and at 2 and 6 weeks. RESULTS: All animals survived the procedure. Pericardial effusion (PE) during the procedure was hemodynamically significant in four of the eight animals. At necropsy, lead exit sites appeared to heal without complication at 2 and 6 weeks. Volume of pericardial fluid was 10.8 +/- 6.2 mL and appeared normal in four of the six chronic animals. Moderate fibrinous deposition was observed in two animals, which had exhibited significant over-procedural PE. CONCLUSIONS: Access to the PS via a transvenous approach is feasible. Pacing leads can be negotiated into this region. The puncture site heals with the lead in place. Further development should focus on eliminating PE and performing this technique in appropriate heart failure models. PMID- 16221258 TI - Chronotropic incompetence in patients with an implantable cardioverter defibrillator: prevalence and predicting factors. AB - Chronotropic incompetence (CI), which has not been systematically examined in the ICD patient population, may have implications for device programming. A total of 123 ICD patients were classified into three groups: single-chamber ICD with sinus rhythm, dual-chamber ICD with sinus rhythm, and single-chamber ICD with permanent atrial fibrillation. Heart rate response, maximum oxygen uptake, and oxygen uptake at the anaerobic threshold were measured during treadmill exercise testing. In addition, clinical variables such as antiarrhythmic drug therapy, underlying heart disease, and left-ventricular (LV) ejection fraction were recorded. Of the patients studied, 38% were chronotropically incompetent (47/123). Significant predictors of CI were as follows: presence of a coronary disease (P = 0.036), prior cardiac surgery (P = 0.037), chronic drug therapy with beta-blockers (P = 0.032), administration of amiodarone (P = 0.025), and a combination of these two forms of treatment (P = 0.01). Spiroergometry revealed reduced exercise capacity (P = 0.041) and lessened VO2max (P = 0.034) among chronotropically incompetent patients. A large percentage of ICD patients demonstrates CI with subsequently reduced physical stress tolerance. In light of the DAVID study, we believe that a closer examination of rate-adaptive modes for ICD patients is warranted under enhanced conditions: (1) optimized AV interval programming; (2) utilization of new algorithms to reduce ventricular pacing in combination with rate-adaptive atrial pacing, with the goal of addressing CI while minimizing ventricular pacing; and (3) an optimized upper heart-rate limit. PMID- 16221259 TI - Clinical results of an advanced SVT detection enhancement algorithm. AB - INTRODUCTION: Supraventricular tachycardia (SVT) has many characteristics that are similar to ventricular tachycardia (VT). This presents a significant challenge for the SVT-detection algorithms of an implantable cardioverter defibrillator (ICD). A newly developed ICD, which utilizes a Vector Timing and Correlation algorithm as well as interval-based conventional SVT discrimination algorithms (Rhythm ID), was evaluated in this study. MATERIALS AND METHODS: This study was a prospective, multicenter trial that evaluated 96 patients implanted with an ICD at 21 U.S. centers. All patients were followed at 2 weeks, 1 month, and every 3 months post implant. A manual Rhythm ID reference vector was acquired prior to any arrhythmia induction. During testing, atrial tachyarrhythmias were induced first, followed by ventricular arrhythmia induction. Induced and spontaneous SVT and VT/ventricular fibrillation (VF) episodes recorded during the trial were annotated by physician investigators. RESULTS: The mean age of the patients implanted with an ICD was 67.3 +/- 10.8 years. Eighty-one percent of patients were male. The primary cardiovascular disease was coronary artery disease, and the primary tachyarrhythmia was monomorphic VT. Implementation of the Rhythm ID algorithm did not affect the VT/VF detection time. There were a total of 370 ventricular tachyarrhythmias (277 induced and 93 spontaneous) and 441 SVT episodes (168 induced and 273 spontaneous). Sensitivity for ventricular tachyarrhythmias was 100%, and specificity for SVT was 92% (94% and 91% for induced and spontaneous SVT, respectively). All patients had a successful manual Rhythm ID acquisition prior to atrial tachyarrhythmia induction. At the 1-month follow-up, the Rhythm ID references were updated automatically an average of 167.8 +/- 122.7 times. Stored Rhythm ID references correlated to patients' normally conducted rhythm 100% at 2 weeks, and 98% at 1 month. CONCLUSIONS: The Rhythm ID algorithm achieved 100% sensitivity for VT/VF, and 92% specificity for SVT. The manual and automatic Rhythm ID update algorithms successfully acquired references, and the updated references were highly accurate. PMID- 16221260 TI - Strategies for the safe magnetic resonance imaging of pacemaker-dependent patients. AB - OBJECTIVE: To determine if strategies used to safely scan nonpacemaker-dependent patients could be applied to facilitate safe MRI of pacemaker-dependent patients. INTERVENTIONS: Ten pacemaker-dependent patients underwent a total of 11 MRI scans of the head and neck. Screening, reprogramming VOO or DOO at 60 ppm, and monitoring strategies were used to facilitate MRI. A transmit-receive coil was used and MRI pulse sequences were modified to limit the whole-body specific absorption rate (SAR). RESULTS: All scans proceeded uneventfully. No difficulties in post-MRI telemetry or interrogation were seen and no post-MRI programming changes were noted. No patient experienced arrhythmia or symptoms during or immediately after MRI. Battery status remained unchanged. No patient experienced post-MRI change in sensing thresholds. Three patients showed no change in the atrial or ventricular pacing thresholds when the pre-MRI values were compared to the immediate post-MRI values and the 3-month follow-up values. All other patients showed a rise or fall of 0.5 V in their chamber threshold values when the pre-MRI, post-MRI, and 3-month follow-up values were compared. More patients showed a fall in their pacing thresholds than a rise post-MRI. CONCLUSION: While clearly a higher risk group, like nonpacemaker-dependent patients, MRI might be performed in pacemaker-dependent patients if appropriate pacemaker reprogramming, patient monitoring, and MRI scanning techniques are implemented. PMID- 16221261 TI - Recurrence of atrial tachyarrhythmias in implantable cardioverter-defibrillator recipients. AB - BACKGROUND: Because the natural history of atrial tachyarrhythmia (AT) is not known in patients with implantable cardioverter-defibrillators (ICDs) but without device-based atrial therapies, we aimed to describe the characteristics and recurrence of AT in such patients. METHODS: In this multicenter trial, 269 patients with standard indications for ICD placement and 2 episodes of AT in the preceding year received a dual-chamber ICD capable of logging AT. Patients were randomly assigned to 3-month periods of atrial therapies "on" or "off." This analysis considered only the 118 patients with atrial therapies programmed off at ICD placement. RESULTS: Fifty-eight patients (49%) had at least 1 AT episode longer than 1 minute, and 21 (18%) had at least 1 prolonged episode (>24 hours). The median episode frequency for each patient (episodes per month) was 1.8 episodes longer than 1 minute, 0.8 longer than 1 hour, and 0 longer than 24 hours. The median AT burden was 12.2 hours per month. CONCLUSIONS: Patients with standard ICD indications and history of AT have infrequent episodes, frequent short episodes, or prolonged episodes of AT-atrial fibrillation. However, the clinical characteristics examined did not distinguish among the groups. Improved diagnostic tools may help identify patients at risk for development of AT, thereby allowing specific therapies to be targeted to each group of patients. PMID- 16221262 TI - Junctional rhythm--a suitable surrogate endpoint in catheter ablation of atrioventricular nodal reentry tachycardia? AB - INTRODUCTION: Current AHA/ACC guidelines state that junctional rhythm (JR) is an acceptable endpoint in patients undergoing radiofrequency ablation (RFA) for narrow complex tachycardia in the presence of dual AV nodal physiology, but in the absence of inducible AVNRT. Only limited data are available on the utility of JR as a marker of successful slow pathway ablation. We sought to further characterize the sensitivity, specificity, and predictive value of JR in AVNRT ablation. METHODS: A retrospective analysis was performed of 387 consecutive patients with documented narrow complex tachycardia referred for ablation, with dual AV nodal physiology and inducible AVNRT at electrophysiological study. RFA of slow pathway was performed, with the presence or absence of JR recorded for each application and inducibility tested using atrial stimulation protocol and isoproterenol. RESULTS: Successful ablation was achieved in 385 of 387 patients using a total of 1,861 applications of radiofrequency energy. JR occurred in 692 applications, giving a sensitivity and specificity of JR as an indicator of successful ablation of 99.5% and 79.1% and a positive predictive value of 55.5%. CONCLUSIONS: This study confirms that successful ablation of slow pathway seldom occurs in the absence of JR. Although JR almost invariably occurs with successful ablation, its lack of specificity and low positive predictive value questions the use of it as an endpoint in AVNRT ablations, and the guidelines should reflect this. PMID- 16221263 TI - Severe left ventricular systolic dysfunction increases atrial fibrillation after ablation of atrial flutter. AB - BACKGROUND: Atrial fibrillation (Afib) that occurs after a successful atrial flutter (AFL) ablation may negate the potential benefits of the ablation. Afib occurs more often when severe left ventricular systolic dysfunction (LVSD) is present. We hypothesized that even after a successful AFL ablation, the incidence of postablation Afib is increased when severe LVSD is present. METHODS: Ninety consecutive patients with LVSD who underwent ablation for AFL at Montefiore Medical Center from August 2001 to January 2005 were classified according to the severity of LVSD. Group 1 (n = 36) consisted of patients with EF < or = 35%, and group 2 (n = 54) consisted of patients with EF 36-55%. There were no statistically significant differences in baseline patient characteristics between the two groups. RESULTS: During a mean follow up of 350 days, Afib occurred in 31% (n = 11; 8 with prior history of AFib) in group 1, and 7.4% (n = 4; all with prior history of Afib) in group 2. Cumulative probability of remaining Afib-free in group 1 versus group 2 was 75% versus 96% at 365 days, and 69% versus 91% at 600 days (P = 0.01). A prior history of Afib did not interact with EF when analyzed with a logistic regression analysis. CONCLUSION: After an AFL ablation, the incidence of Afib is increased, and the probability of remaining free of Afib is decreased, when severe LVSD is present, independent of a prior history of Afib. This finding may have implications for optimal patient selection for AFL ablation, and the use of adjunctive therapies. PMID- 16221264 TI - Artificial attenuation of ECG voltage produces shortening of the corresponding QRS duration: clinical implications for patients with edema. AB - BACKGROUND: Prolonged QRS duration (QRSd) is a useful index for the management of patients with congestive heart failure (CHF). QRSd is affected by changes in the ECG voltage (ECGV) in the context of development and amelioration of peripheral edema (PERE), independent of underlying pathology. Nowadays, physicians accept QRSd measured by computer techniques. The latter offers the possibility of testing the hypothesis that artificial alteration of the ECGV, simulating effects of PERE, could lead to changes in the QRSd. METHODS: To this end, voltage was attenuated by 25%, 50%, and 75% in 100 digital ECGs recorded from normal subjects and in 20 patients with complete left bundle branch block (LBBB), by merely increasing the calibration strength by 4/3, 2, and 4, respectively, and by using the same data. RESULTS: All ECGs were analyzed by the same computer program and this led to a reduction of global QRSd by 2.3 +/- 2.9%, 5.7 +/- 4.0%, and 11.9 +/ 6.2%, respectively, in the normal subjects, and 1.6 +/- 1.4%, 3.4 +/- 1.7%, and 8.2 +/- 3.6%, respectively, in the patients with LBBB. Correlation of the percent change in the global QRSd and the percent change in ECGV was good with an r = 0.65, and P = 0.00005 in the normal subjects, and an r = 0.74 and P = 0.00005 in the patients with LBBB. CONCLUSIONS: Apparent shortening in QRSd as a function of ECGV attenuation due to PERE could have implications in the follow-up of patients with CHF, and their selection for implantable cardioverter/defibrillators, or cardiac resynchronization therapy. PMID- 16221265 TI - Electrophysiologic characteristics of atria in patients without heart disease. AB - BACKGROUND: The significance of atrial fibrillation or tachycardia (AF) induction remains debatable. Some believe that the presence of heart disease (HD) increases the sensitivity and decreases the specificity of programmed atrial stimulation (PAS). There are few data in patients without HD. The purpose of the study was to evaluate the results of PAS in asymptomatic patients without HD and in those with documented spontaneous AF, but without HD, to know the diagnosis value of the technique. METHODS: A total of 4,900 PAS were consecutively performed. The control group (I, N=67) was defined by the absence of preexcitation syndrome, dizziness/syncope, hypertension, history of tachycardia, or other documented HD together with a normal 2D echocardiogram and 24-hour Holter monitoring. They were compared to a group (II) of 54 patients with documented paroxysmal AF and without HD. PAS used one and two extrastimuli, delivered during sinus rhythm and two drive rates (600, 400 ms). Atrial-effective refractory periods (ARP), their adaptation to cycle length, and conduction times were noted. AF induction was defined as the induction of AF lasting more than 1 minute. RESULTS: Group I patients (1.4% of 4,900) were younger than group II (51 +/- 17 vs 65 +/- 11 years, P < 0.001). A single extrastimulus never induced sustained AF in group I, but did so in 11 group II patients (20%); sustained AF was induced by two extrastimuli in 15 group I patients (22%) and in 31 group II patients (57%) (P < 0.001). There were no ARP and conduction time differences in group I patients with and without inducible AF, but there was a longer sinus cycle length in patients with inducible AF (977 +/- 164 vs 838 +/- 141 ms, P < 0.02). There were no electrophysiological differences in group II patients with and without inducible AF. No group I patient developed spontaneous AF (follow-up 4 +/- 2 years). The sensitivity of PAS with one extrastimulus was 20% and the specificity 100%; the sensitivity of PAS with two extrastimuli was 57% and the specificity 78%. CONCLUSION: Sustained AF was not induced by one extrastimulus in control patients without symptoms, nor heart disease, but sustained AF was induced by two extrastimuli in 22% of these patients. The induction of a sustained AF by two extrastimuli should be interpreted cautiously, particularly in patients with a relative sinus bradycardia. However, the sensitivity of PAS with one extrastimulus was very low and two extrastimuli were required in patients with spontaneous AF to induce the tachycardia. Other electrophysiological parameters were not useful to differentiate patients with and without inducible AF. PMID- 16221266 TI - Magnetic resonance angiography of pulmonary veins: implications for catheter ablation of atrial fibrillation. AB - INTRODUCTION: Catheter ablation of atrial fibrillation (AF) requires exact anatomical information about pulmonary venous (PV) ostia. In this study, anatomy of pulmonary veins (PVs) was assessed using three-dimensional (3D) reconstructions of magnetic resonance angiography (MRA). METHODS AND RESULTS: Contrast-enhanced MRA of the PVs was performed in 40 patients (mean age 53 +/- 9 years) with paroxysmal (30 patients) or persistent (10 patients) AF, scheduled for circumferential ablation around PV ostia. PV ostial anatomy and diameters were evaluated from multiplanar reconstructions and compared with 3D reconstructions. Thirty (75%) patients presented with a common left-sided antrum (21 short and 9 long trunk), while additional PVs were found on right side in 23%. PV ostia were oblong in shape (mean diameters 17.0 +/- 4.3 vs 10.5 +/- 2.5 mm by two-dimensional (2D) measurements, and 20.8 +/- 7.6 mm vs 12.9 +/- 3.3 mm by 3D reconstruction, in long and short axis, respectively). There was a correlation between measurements obtained from 2D and 3D images, although 3D imaging provided slightly larger diameters. CONCLUSIONS: MRA with 3D reconstructions is an important technique for preprocedural assessment of PVs that allows full understanding of their anatomy and size. This information may be important for selection of appropriate tools. PMID- 16221267 TI - Comparison of the efficacy of cooled-tip and 8-mm-tip catheters for radiofrequency catheter ablation of the cavotricuspid isthmus: a meta-analysis. AB - BACKGROUND: Cooled-tip RFA (cRFA) and conventional 8-mm-tip catheters were found to be more effective and as safe as conventional 4-mm-tip catheters for atrial flutter (AFL) radiofrequency ablation (RFA), facilitating the rapid achievement of bi-directional isthmus block (BIB), but studies comparing cRFA and 8-mm-tip catheters are not randomized or results are discussed. Thus, we performed a meta analysis of available randomized trials to evaluate the effectiveness in terms of primary success and procedure parameters. METHODS: Reports of trials were identified through a Medline, Embase, Current Contents, Cardline, and an extensive bibliography search. Trials that met the following criteria were included: (1) prospective, randomized, controlled, and open trials; (2) patients assigned to an 8-mm-tip or a cRFA catheter for AFL RFA; (3) endpoints events related to primary success rate (BIB achievement), and procedure parameters (number of RF applications, x-ray exposure and ablation duration). RESULTS: Seven trials met the inclusion criteria. They included 603 patients with established AFL randomized to an 8-mm-tip or cRFA catheter group. Comparing 8-mm groups with cRFA groups, the meta-analysis showed similar BIB achievement relative risk (RR) 0.96, 95% confidence interval (CI): 0.92 to 1.01, (P = 0.13); total RF application time weighted mean difference (WMD) 0.88, 95% CI: -0.36 to 2.12, (P = 0.16); duration of x-ray exposure (min) (WMD = 1.07, 95% CI: -0.81 to 0.295, (P = 0.26); ablation procedure duration (min) (WMD = 0.68, 95% CI: -3.37 to 4.73; P = 0.74). CONCLUSION: The present meta-analysis confirms that cooled-tip and large tip catheters are equally efficient for cavotricuspid isthmus ablation with both similar primary success rates and procedure parameters. PMID- 16221268 TI - Automated ventricular substrate mapping--evaluation in an ovine chronic myocardial infarction model. AB - INTRODUCTION: We hypothesized that automated electrogram analysis might enable rapid localization of ventricular scar. This would allow the delivery of interventions such as radiofrequency ablation or therapeutic agents to critical areas within the scar and scar periphery. METHODS: Substrate mapping was performed on seven sheep 36.5 +/- 32.9 weeks after a left anterior descending artery myocardial infarction had been induced. Contact electrograms and the mapping catheter three-dimensional (3D) location were recorded simultaneously. A computer program was written in-house to automatically identify sinus beats, analyze electrogram characteristics (e.g., electrogram amplitude and minimum slope), and integrate the analysis results into a 3D scar map. RESULTS: The total time required to produce the scar maps was a mean of 8.3 +/- 2.0 minutes. The automated substrate mapping (ASM) system beat detection algorithm had a high sensitivity (i.e., detected 87.4% of the recorded beats) and excellent specificity (only one false activation over 58.2 minutes of total recorded data). The system was able to classify the detected beats ('sinus' or 'ectopic') with high specificity (specificity = 97.3% confidence interval (CI): 96.9-97.7) and moderate sensitivity (sensitivity = 78.3% CI: 77.3%-79.5%). The scar area identified by the ASM system correlated well with the pathologically defined scar area (R2 = 0.87 p < 0.001). CONCLUSIONS: ASM enables accurate scar maps to be produced rapidly. This strategy may play an important role for both clinical and research applications, allowing therapeutic agents and radiofrequency ablation to be delivered to critical locations in and around ventricular scar. PMID- 16221269 TI - Left ventricular epicardial activation increases transmural dispersion of repolarization in healthy, long QT, and dilated cardiomyopathy dogs. AB - BACKGROUND: Benefits of cardiac resynchronization therapy (CRT) are well established. However, less is understood concerning its effects on myocardial repolarization and the potential proarrhythmic risk. METHODS AND RESULTS: Healthy dogs (n = 8) were compared to a long QT interval (LQT) model (n = 8, induced by cesium chloride, CsCl) and a dilated cardiomyopathy with congestive heart failure (DCM-CHF, induced by rapid ventricular pacing, n = 5). Monophasic action potential (MAP) recordings were obtained from the subendocardium, midmyocardium, subepicardium, and the transmural dispersion of repolarization (TDR) was calculated. The QT interval and the interval from the peak to the end of the T wave (T(p-e)) were measured. All these characteristics were compared during left ventricular epicardial (LV-Epi), right ventricular endocardial (RV-Endo), and biventricular (Bi-V) pacing. In healthy dogs, TDR prolonged to 37.54 ms for Bi-V pacing and to 47.16 ms for LV-Epi pacing as compared to 26.75 ms for RV-Endo pacing (P < 0.001), which was parallel to an augmentation in T(p-e) interval (Bi V pacing, 64.29 ms; LV-Epi pacing, 57.89 ms; RV-Endo pacing, 50.29 ms; P < 0.01). During CsCl exposure, Bi-V and LV-Epi pacing prolonged MAPD, TDR, and T(p-e) interval as compared to RV-Endo pacing. The midmyocardial MAPD (276.30 ms vs 257.35 ms, P < 0.0001) and TDR (33.80 ms vs 27.58 ms, P=0.002) were significantly longer in DCM-CHF dogs than those in healthy dogs. LV-Epi and Bi-V pacing further prolonged the MAPD and TDR in this model. CONCLUSIONS: LV-Epi and Bi-V pacing result in prolongation of ventricular repolarization time, and increase of TDR accounted for a parallel augmentation of the T(p-e) interval, which provides evidence that T(p-e) interval accurately represents TDR. These effects are magnified in the LQT and DCM-CHF canine models in addition to their intrinsic transmural heterogeneity in the intact heart. This mechanism may contribute to the development of malignant ventricular arrhythmias, such as torsades de pointes (TdP) in congestive heart failure (CHF) patients treated with CRT. PMID- 16221270 TI - Mechanism of transition from irregular to regular supraventricular tachycardia. PMID- 16221271 TI - Changes in intrathoracic impedance from a pneumothorax: insights from an implanted monitoring system. AB - The measurement of transthoracic impedance is now possible using new implantable cardioverter-defibrillators. This can be used to monitor fall in impedance associated with increasing pulmonary oedema. We describe a case of a large rapid increase of impedance and dyspnoea related to a pneumothorax. PMID- 16221272 TI - Inappropriate sinus tachycardia, postural orthostatic tachycardia syndrome, and overlapping syndromes. AB - BACKGROUND: Inappropriate sinus tachycardia (IAST) and postural orthostatic tachycardia syndrome (POTS) are syndrome complexes with some distinctive features, overlapping clinical manifestations, and potential common mechanisms. Pathogenesis of these overlapping syndromes is poorly understood. Diagnostic and therapeutic approaches have not been standardized. PURPOSE: This article provides an overview of the definition, clinical presentation, and proposed mechanisms of IAST and other overlapping syndromes. A stepwise diagnostic approach is suggested. A multidisciplinary management scheme is outlined. METHODS: A MEDLINE search for English-language articles on IAST, POTS, and chronic orthostatic intolerance published up to 2005 was performed. Published data incorporated with our clinical experience were synthesized and presented in this review. RESULTS: The population of IAST is heterogeneous and underlying mechanisms are complex and likely multifactorial. Evidence suggests that both cardiac and extracardiac causes are plausible. Regional and limited autonomic neuropathies, at least in part, can provide a mechanism-based explanation of the cardiovascular indices and clinical symptoms in a significant number of patients with IAST. The regional abnormalities can be detected by autonomic testing. Among patients with IAST and evidence of autonomic dysregulation, an integrated autonomic, cardiovascular, and psychiatric management approach appears to be logical and rational when appropriate. Sinus node ablation could be considered in patients with persistent IAST in the absence of autonomic neuropathy and multisystem symptoms. Data from long-term outcomes are lacking. CONCLUSION: The current understanding of IAST mechanisms is incomplete and management approach is not adequate. Significant effort needed in clinical research to improve therapeutic outcome. PMID- 16221273 TI - Pacing to restore right ventricular contraction after surgical disconnection for arrhythmia control in right ventricular cardiomyopathy. AB - Ventricular tachycardia in ARVC (arrhythmogenic right ventricular cardiomyopathy) is typically managed by ICD implantation, with a limited role of catheter ablation. Surgical disconnection of the right ventricular (RV) has been used to control ventricular tachycardia (VT) in ARVC, but it often led to refractory RV failure due to loss of RV contraction after surgery. We report multisite pacing to recruit the disconnected RV to prevent ventricular failure. PMID- 16221274 TI - Right heart failure due to loss of right ventricular capture in a patient with atrioventricular junction ablation and biventricular pacing. AB - We describe the case of a patient with atrioventricular (AV) junction ablation and chronic biventricular pacing in which intermittent dysfunction of the right ventricular (RV) lead resulted in left ventricular (LV) stimulation alone and onset of severe right heart failure. Restoration of biventricular pacing by increasing device output and then performing lead revision resolved the issue. This case provides evidence that LV pacing alone in patients with AV junction ablation may lead to severe right heart failure, most likely as a result of iatrogenic mechanical dyssynchrony within the RV. Thus, probably this pacing mode should be avoided in pacemaker-dependent patients with heart failure. PMID- 16221275 TI - Contralateral replacement of pacemaker and leads following laser sheath extraction and concomitant stenting for superior vena cava syndrome. AB - We present the case of a 61-year-old man with a history of pacemaker implantation 15 years ago. The patient was admitted with chronic swelling of the right hemithorax and inflammation of the old incision scar after the replacement of pacemaker generator 6 years ago. The patient also presented symptoms and clinical signs of superior vena cava syndrome. Computed tomography of the thorax showed obstruction of the superior vena cava. The surgical procedure consisted of extraction of infected pacemaker system with excimer laser technique followed by dilatation and stenting of the SVC and finally implantation of a new permanent pacemaker system on the left side. PMID- 16221276 TI - Successful pacemaker lead implantation via the subclavian vein despite of high degree stenosis of the vein. AB - This case reports on an 81-year-old man with implanted dual-chamber pacemaker for binodal disease who required ventricular lead revision due to loss of ventricular capture. Successful placement of a new lead via the subclavian vein despite of high degree stenosis of the medial part of the vein is described using a very thin bipolar ventricular pacing lead. Currently available pacing electrodes for this purpose are reviewed. PMID- 16221277 TI - Anatomic and electrophysiologic evaluation of a right lateral atrioventricular Mahaim fiber. AB - We report a patient who underwent an electrophysiologic study and radiofrequency catheter ablation for a right lateral Mahaim fiber. During sinus rhythm with overt preexcitation, propagation mapping was performed in the right ventricle using a three-dimensional electro-anatomical mapping system (CARTO). Small discrete potentials, which reflected the excitation of the Mahaim fiber, could be recorded along the line from the vicinity of the parental tricuspid annulus to approximately one-third of distal site from the base to the apex. The relationship of the timing of its potential to the anatomical location could be disclosed on recordings of the local electrogram and anatomical map. PMID- 16221278 TI - Ventricular tachycardias and dilated cardiomyopathy caused by Fabry disease. AB - Women with Fabry disease are considered to have a good prognosis. Here we report the clinical history of a female patient with severe Fabry disease before and after referral to our hospital. She was diagnosed to have a cardiac variant of Fabry disease since a left ventricular endomyocardial biopsy revealed specific features of this disease and that cardiac involvement was the sole manifestation. She developed sustained ventricular tachycardia and dilated cardiomyopathy requiring oral administration of amiodarone and a cardiac defibrillator implant. PMID- 16221281 TI - T-wave alternans using ventricular and atrial pacing. PMID- 16221282 TI - Chronic inflammatory demyelinating polyradiculoneuropathy: new views and guidelines. PMID- 16221284 TI - A critical review of interfaces with the peripheral nervous system for the control of neuroprostheses and hybrid bionic systems. AB - Considerable scientific and technological efforts have been devoted to develop neuroprostheses and hybrid bionic systems that link the human nervous system with electronic or robotic prostheses, with the main aim of restoring motor and sensory functions in disabled patients. A number of neuroprostheses use interfaces with peripheral nerves or muscles for neuromuscular stimulation and signal recording. Herein, we provide a critical overview of the peripheral interfaces available and trace their use from research to clinical application in controlling artificial and robotic prostheses. The first section reviews the different types of non-invasive and invasive electrodes, which include surface and muscular electrodes that can record EMG signals from and stimulate the underlying or implanted muscles. Extraneural electrodes, such as cuff and epineurial electrodes, provide simultaneous interface with many axons in the nerve, whereas intrafascicular, penetrating, and regenerative electrodes may contact small groups of axons within a nerve fascicle. Biological, technological, and material science issues are also reviewed relative to the problems of electrode design and tissue injury. The last section reviews different strategies for the use of information recorded from peripheral interfaces and the current state of control neuroprostheses and hybrid bionic systems. PMID- 16221283 TI - European Federation of Neurological Societies/Peripheral Nerve Society Guideline on management of chronic inflammatory demyelinating polyradiculoneuropathy. Report of a joint task force of the European Federation of Neurological Societies and the Peripheral Nerve Society. AB - BACKGROUND: Numerous sets of diagnostic criteria have sought to define chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), and randomized trials and systematic reviews of treatment have been published. OBJECTIVES: The aim of this guideline was to prepare consensus guidelines on the definition, investigation, and treatment of CIDP. METHODS: Disease experts and a representative of patients considered references retrieved from MEDLINE and Cochrane Systematic Reviews in May 2004 and prepared statements that were agreed in an iterative fashion. RECOMMENDATIONS: The Task Force agreed on good practice points to define clinical and electrophysiological diagnostic criteria for CIDP with or without concomitant diseases and investigations to be considered. The principal treatment recommendations were as follows: (1) intravenous immunoglobulin (IVIg) or corticosteroids should be considered in sensory and motor CIDP (level B recommendation); (2) IVIg should be considered as the initial treatment in pure motor CIDP (good practice point); (3) if IVIg and corticosteroids are ineffective, plasma exchange should be considered (level A recommendation); (4) if the response is inadequate or the maintenance doses of the initial treatment are high, combination treatments or adding an immunosuppressant or immunomodulatory drug should be considered (good practice point); and (5) symptomatic treatment and multidisciplinary management should be considered (good practice point). PMID- 16221285 TI - Multicenter study of peroneal mononeuropathy: clinical, neurophysiologic, and quality of life assessment. AB - This is a multicenter study on peroneal mononeuropathy (PM), in which a multidimensional protocol was performed to evaluate (1) the predisposing factors and their occurrence; (2) the relationships between the etiological, clinical, and neurophysiologic findings; and (3) disability and quality of life (QoL) in a wide sample with PM. Clinical and neurophysiologic evaluation was performed in all patients; moreover, the group adopted validated disability and QoL measurements to obtain more comprehensive and reliable data on PM. From November 2002 to January 2004, 69 patients were enrolled consecutively in 11 Italian centers. Our data showed that PM involves men more frequently than women (male : female = 4.1:1). PM was idiopathic (16%) or due to prolonged posture (23.1%), surgery (20.3%), weight loss (14.5%), trauma (11.6%), bedridden condition (7.3%), external compression from cast (5.8%), and arthrogenic cyst at the fibula (1.4%). Unexpectedly, peroneal nerve lesions were not only due to surgical operation close to the peroneal region but were also associated with thoracic-abdominal surgery. We observed conduction block in about 50-70% of postural and weight loss PM; in perioperative and idiopathic PM, conduction block or mixed damage was equally present; in PM due to trauma, we observed an exclusive axonal damage in about 60% of cases. Only in three cases (one postural PM, one idiopathic PM, and one weight loss PM), we observed a slowing of conduction velocity in the popliteal fossa-fibular head segment without conduction block. The comparison between QoL in patients with PM and in healthy subjects showed a significant involvement of physical and mental aspects. With regard to disability, 68% of patients walked with difficulty. Our data show that (1) most of the cases of PM are due to an identifiable predisposing factor; (2) there is a good correlation between predisposing factors and clinical-neurophysiologic findings; and (3) PM causes disability and deterioration of the physical and emotional aspects of QoL. PMID- 16221286 TI - Influence of aging on thermal and vibratory thresholds of quantitative sensory testing. AB - Quantitative sensory testing has become a common approach to evaluate thermal and vibratory thresholds in various types of neuropathies. To understand the effect of aging on sensory perception, we measured warm, cold, and vibratory thresholds by performing quantitative sensory testing on a population of 484 normal subjects (175 males and 309 females), aged 48.61 +/- 14.10 (range 20-86) years. Sensory thresholds of the hand and foot were measured with two algorithms: the method of limits (Limits) and the method of level (Level). Thresholds measured by Limits are reaction-time-dependent, while those measured by Level are independent of reaction time. In addition, we explored (1) the correlations of thresholds between these two algorithms, (2) the effect of age on differences in thresholds between algorithms, and (3) differences in sensory thresholds between the two test sites. Age was consistently and significantly correlated with sensory thresholds of all tested modalities measured by both algorithms on multivariate regression analysis compared with other factors, including gender, body height, body weight, and body mass index. When thresholds were plotted against age, slopes differed between sensory thresholds of the hand and those of the foot: for the foot, slopes were steeper compared with those for the hand for each sensory modality. Sensory thresholds of both test sites measured by Level were highly correlated with those measured by Limits, and thresholds measured by Limits were higher than those measured by Level. Differences in sensory thresholds between the two algorithms were also correlated with age: thresholds of the foot were higher than those of the hand for each sensory modality. This difference in thresholds (measured with both Level and Limits) between the hand and foot was also correlated with age. These findings suggest that age is the most significant factor in determining sensory thresholds compared with the other factors of gender and anthropometric parameters, and this provides a foundation for investigating the neurobiologic significance of aging on the processing of sensory stimuli. PMID- 16221287 TI - Optimizing the use of electrophysiology in the diagnosis of chronic inflammatory demyelinating polyneuropathy: a study of 20 cases. AB - Current electrophysiologic criteria for chronic inflammatory demyelinating polyneuropathy (CIDP) are highly specific but poorly sensitive. The required extensiveness and best practical way of performing nerve conduction studies to achieve optimal sensitivity remain unknown. We here initially retrospectively analyzed the motor nerve conduction study results of 20 consecutive patients with a clinical diagnosis of CIDP (four performed prior to, and 16 after, treatment initiation) to assess the sensitivity of six published sets of criteria (Nicolas et al., 2002; Thaisetthawatkul et al., 2002; Ad Hoc Subcommittee of the American Academy of Neurology AIDS Taskforce, 1991; Magda et al., 2003; Hughes et al., 2001; Saperstein et al., 2001), as well as four combinations (Nicolas et al., 2002; Ad Hoc Subcommittee of the American Academy of Neurology AIDS Taskforce, 1991; Hughes et al., 2001; Saperstein et al., 2001, each individually combined with Thaisetthawatkul et al., 2002). Sensitivity was highest for the combination of Nicolas et al. (2002) and Thaisetthawatkul et al. (2002) (100%). We then determined the sensitivity of this combined criteria, using five different, hypothetical, nerve conduction study protocols, applied retrospectively to the neurophysiologic data of our 20 patients (exclusive upper limb studies with proximal stimulations; exclusive lower limb studies; full forearm and foreleg studies without proximal stimulations; right-sided studies with proximal stimulations; and left-sided studies with proximal stimulations). The findings showed that exhaustive upper limb or, alternatively, four-limb forearm and foreleg testing would have proved considerably more sensitive than unilateral or lower limb studies to achieve an electrophysiologic diagnosis of CIDP. PMID- 16221288 TI - Different expressions of BDNF, NT3, and NT4 in muscle and nerve after various types of peripheral nerve injuries. AB - The changes in the expression of brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4) in the rat neuromuscular system as a result of three different types of sciatic nerve injuries have been evaluated. The changes in mRNA and protein levels for BDNF, NT-3, and NT-4 in the soleus muscle and sciatic nerve were assessed 4-28 days after sciatic nerve transection (neurotmesis), sciatic nerve crush (axonotmesis), and mild acute compression (neurapraxia). BDNF mRNA levels increased dramatically with nerve transection in the soleus muscle and the sciatic nerve 7-14 days after injury, whereas the changes were low in other types of injury. The changes of protein levels for BDNF were also similar. The mRNA and the protein levels of NT-3 in the soleus muscle did not show any significant difference. The mRNA for NT-4 in the soleus muscle decreased from 4 to 14 days after sciatic nerve transection, and the protein level was also minimum 14 days after sciatic nerve transection. Our results indicate that the neurotrophic factors in the neuromuscular system could play a role in differentiating peripheral nerve injury. PMID- 16221289 TI - Patterns of P-glycoprotein activity in the nervous system during vincristine induced neuropathy in rats. AB - Vincristine (VCT) is a neurotoxic agent and also a substrate of multidrug resistance (MDR) transporters such as P-glycoprotein (P-gp) and MDR-associated proteins 1 and 2 (MRP1 and MRP2). These proteins are expressed in the central and peripheral nervous systems (CNS and PNS) and normally protect these structures against the harmful effects of VCT. The aim of this study was to elucidate the paradoxical relation between the MDR transporters and the VCT neurotoxicity. With a validated rat model of VCT-induced neuropathy, (1) the expressions of mdr1a (P gp), mdr1b (P-gp), mrp1 (MRP1), and mrp2 (MRP2) genes were assessed by quantitative real-time polymerase chain reaction, and (2) the transporter activity was monitored using a radioactive tracer, (99m)Tc-sestamibi, in the CNS and PNS. The results showed higher expression of mdr1a and mdr1b genes (x3 and x35, respectively) in the brain than in the spinal ganglia in both control and treated animals. Transporter activity was higher (x10) in the CNS than in the PNS. Hence, P-gp protection may be lower in the PNS than in the CNS, and this may be responsible for the peripheral neurotoxicity of P-gp substrates. VCT treatment increased expression of the mdr1a gene in the CNS and PNS (both x1.7), mrp1 gene in the PNS (x1.7), and transporter activity in both the CNS and the PNS (x4 and x8, respectively). This transporter induction may induce adverse effects when analgesic drugs are administered to treat neuropathic pain. PMID- 16221290 TI - Tumor necrosis factor-alpha-converting enzyme is expressed in the inflamed peripheral nervous system. AB - Tumor necrosis factor-alpha (TNF-alpha) is considered to play a critical role in the pathogenesis of immune-mediated inflammatory demyelinating disorders of the peripheral nervous system (PNS). Processing of membrane-bound inactive pro-TNF alpha into the active soluble cytokine is mediated by a sheddase, the so-called TNF-alpha-converting enzyme (TACE), a member of the A Disintegrin and Metalloproteinase (ADAM) family. We explored the expression of TACE (ADAM-17) in sciatic nerves from Lewis rats with experimental autoimmune neuritis (EAN), an animal model of the Guillain-Barre syndrome (GBS), an immune-mediated polyradiculoneuropathy. To extend our study to human disease, sural nerve biopsies from GBS patients were investigated by immunohistochemistry. In EAN, T lymphocytes could be defined as the cellular source of ADAM-17 with peak expression levels at maximum clinical disease severity. Similarly, in human sural nerves, ADAM-17-expressing T cells could be localized primarily within the epi- and perineurium, whereas in control sections from patients with non-inflammatory neuropathies, no expression could be depicted. Our findings indicate that ADAM-17 is upregulated during EAN and expressed in nerves of GBS patients and thus may contribute to the pathogenesis of inflammatory demyelination of the PNS. PMID- 16221291 TI - Agmatine treatment and vein graft reconstruction enhance recovery after experimental facial nerve injury. AB - The rate of nerve regeneration is a critical determinant of the degree of functional recovery after injury. Here, we sought to determine whether treatment with the neuroprotective compound, agmatine, with or without nerve reconstruction utilizing a regional autogenous vein graft would accelerate the rate of facial nerve regeneration. Experiments compared the following seven groups of adult male rats: (A) Intact untreated controls. (B) Sham operation with interruption of the nerve blood supply (controls). (C) Transection of the mandibular branch of the facial nerve (generating a gap of 3 mm) followed by saline treatment. (D) Nerve transection with unsutured autogenous vein (external jugular) graft reconstruction plus saline treatment. (E) Nerve transection with sutured vein graft approximation (coaptation of the proximal and distal nerve stumps) plus saline. (F) Nerve transection with sutured vein graft followed by agmatine treatment (four daily intraperitoneal injections of 100 mg/kg agmatine sulfate). (G) Nerve transection with unsutured vein graft followed by agmatine treatment. Functional recovery, as assessed by grading vibrissae movements and by recording nerve conduction velocity and numbers of regenerated axons, indicated that either vein reconstruction or agmatine treatment resulted in accelerated and more complete recovery as compared with controls. But best results were observed in animals that underwent combined treatment, i.e., vein reconstruction plus agmatine injection. We conclude that agmatine treatment can accelerate facial nerve regeneration and that agmatine treatment together with autogenous vein graft offers an advantageous alternative to other facial nerve reconstruction procedures. PMID- 16221292 TI - Fatigue as the main presenting symptom of chronic inflammatory demyelinating polyradiculoneuropathy: a study of 11 cases. AB - Fatigue has been shown to be more frequent than previously thought in immune mediated polyneuropathies. However, fatigue has not been reported as the main cause of referral in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) patients. Between January 2001 and December 2003, we investigated 11 patients referred for fatigue, for which we established a final diagnosis of CIDP. All patients had at least two clinical examinations including assessment of the fatigue severity scale (FSS) and one electrophysiological and laboratory work up. Additionally, 10 of the 11 patients had a nerve biopsy. There were 11 male patients. Mean age at onset was 53 +/- 11 years. Main cause of referral was fatigue in all patients. Additional symptoms included cramps (one case), distal paresthesias (six cases), limb pain (seven cases) and vasomotor disturbances (one case). Cerebrospinal fluid (CSF) analysis displayed a moderate increase in protein content in four patients. Electrophysiological analysis showed abnormalities in all patients. Among 11 patients, one fulfilled the American Academy of Neurology electrodiagnostic criteria for CIDP and three fulfilled the inflammatory neuropathy cause and treatment group or the Nicolas et al. criteria. In the eight remaining patients, a nerve biopsy confirmed the diagnosis of CIDP. Ten patients were treated, among which seven showed a significant improvement based on the FSS scale. This study shows that fatigue is a possible cause of referral for patients with CIDP and, like previous reports, emphasizes the lack of sensitivity of widely accepted electrophysiological criteria of CIDP. Long term follow up of these patients is warranted to determine the prognosis of these minimal forms of CIDP and establish the best therapeutic strategy in such cases. PMID- 16221293 TI - Carpal tunnel syndrome or congenital hand anomaly: a clinical and electromyographic study. PMID- 16221294 TI - Guillain-Barre syndrome with optic neuritis and cytomegalovirus infection. PMID- 16221295 TI - Utility of dorsal sural nerve in early determination of diabetic polyneuropathy. PMID- 16221296 TI - A need to clarify the role of apolipoprotein E in peripheral nerve injury and repair. PMID- 16221297 TI - Oral infection with the Salmonella enterica serovar Gallinarum 9R attenuated live vaccine as a model to characterise immunity to fowl typhoid in the chicken. AB - BACKGROUND: Salmonella enterica serovar Gallinarum (S. Gallinarum) is the causative agent of fowl typhoid, a severe systemic disease of chickens that results in high mortality amongst infected flocks. Due to its virulence, the immune response to S. Gallinarum is poorly characterised. In this study we have utilised infection by the live attenuated S. Gallinarum 9R vaccine strain in inbred chickens to characterise humoral, cellular and cytokine responses to systemic salmonellosis. RESULTS: Infection with 9R results in a mild systemic infection. Bacterial clearance at three weeks post infection coincides with increases in circulating anti-Salmonella antibodies, increased T cell proliferation to Salmonella challenge and increased expression of interferon gamma. These responses peak at four weeks post infection, then decline. Only modest increases of expression of the pro-inflammatory cytokine interleukin-1beta were detected early in the infection. CONCLUSION: Infection of chickens with the 9R vaccine strain induces a mild form of systemic salmonellosis. This induces both cellular and humoral immune responses, which peak soon after bacterial clearance. Unlike enteric-associated Salmonella infections the immune response is not prolonged, reflecting the absence of persistence of Salmonella in the gastrointestinal tract. The findings here indicate that the use of the S. Gallinarum 9R vaccine strain is an effective model to study immunity to systemic salmonellosis in the chicken and may be employed in further studies to determine which components of the immune response are needed for protection. PMID- 16221298 TI - T.I.M.S: TaqMan Information Management System, tools to organize data flow in a genotyping laboratory. AB - BACKGROUND: Single Nucleotide Polymorphism (SNP) genotyping is a major activity in biomedical research. The Taqman technology is one of the most commonly used approaches. It produces large amounts of data that are difficult to process by hand. Laboratories not equipped with a Laboratory Information Management System (LIMS) need tools to organize the data flow. RESULTS: We propose a package of Visual Basic programs focused on sample management and on the parsing of input and output TaqMan files. The code is written in Visual Basic, embedded in the Microsoft Office package, and it allows anyone to have access to those tools, without any programming skills and with basic computer requirements. CONCLUSION: We have created useful tools focused on management of TaqMan genotyping data, a critical issue in genotyping laboratories without a more sophisticated and expensive system, such as a LIMS. PMID- 16221299 TI - Frequent attenders in general practice: problem solving treatment provided by nurses [ISRCTN51021015]. AB - BACKGROUND: There is a need for assistance from primary care mental health workers in general practice in the Netherlands. General practitioners (GPs) experience an overload of frequent attenders suffering from psychological problems. Problem Solving Treatment (PST) is a brief psychological treatment tailored for use in a primary care setting. PST is provided by nurses, and earlier research has shown that it is a treatment at least as effective as usual care. However, research outcomes are not totally satisfying. This protocol describes a randomized clinical trial on the effectiveness of PST provided by nurses for patients in general practice. The results of this study, which currently being carried out, will be presented as soon as they are available. METHODS/DESIGN: This study protocol describes the design of a randomized controlled trial to investigate the effectiveness and cost-effectiveness of PST and usual care compared to usual care only. Patients, 18 years and older, who present psychological problems and are frequent attenders in general practice are recruited by the research assistant. The participants receive questionnaires at baseline, after the intervention, and again after 3 months and 9 months. Primary outcome is the reduction of symptoms, and other outcomes measured are improvement in problem solving skills, psychological and physical well being, daily functioning, social support, coping styles, problem evaluation and health care utilization. DISCUSSION: Our results may either confirm that PST in primary care is an effective way of dealing with emotional disorders and a promising addition to the primary care in the UK and USA, or may question this assumption. This trial will allow an evaluation of the effects of PST in practical circumstances and in a rather heterogeneous group of primary care patients. This study delivers scientific support for this use and therefore indications for optimal treatment and referral. PMID- 16221300 TI - Chronic koro-like symptoms - two case reports. AB - BACKGROUND: Koro is a culture bound syndrome, which has been reported usually from Asian countries. It has been described as an acute, brief lasting illness, which often occurs in epidemics. There is no description in literature of a chronic form of this syndrome. CASE PRESENTATION: Two sporadic cases with koro like symptoms from East India are presented where the illness had a chronic course with durations spanning more than ten years. In contrast to acute, good prognosis, psycho-education responsive form that is usually seen in epidemics; the chronic form, appeared to be associated with greater morbidity and poorer response to interventions. CONCLUSION: There is a possibility of a chronic form of koro syndrome. PMID- 16221302 TI - Laparoscopic repair of high rectovaginal fistula: is it technically feasible? AB - BACKGROUND: Rectovaginal fistula (RVF) is an epithelium-lined communication between the rectum and vagina. Most RVFs are acquired, the most common cause being obstetric trauma. Most of the high RVFs are repaired by conventional open surgery. Laparoscopic repair of RVF is rare and so far only one report is available in the literature. METHODS: We present a case of high RVF repaired by laparoscopy. 56-year-old female who had a high RVF following laparoscopic assisted vaginal hysterectomy was successfully operated laparoscopically. Here we describe the operative technique and briefly review the literature. RESULTS: The postoperative period of the patient was uneventful and after a follow up of 6 months no recurrence was found. CONCLUSION: Laparoscopic repair of high RVF is feasible in selected patients but would require proper identification of tissue planes and good laparoscopic suturing technique. PMID- 16221301 TI - Mammalian BEX, WEX and GASP genes: coding and non-coding chimaerism sustained by gene conversion events. AB - BACKGROUND: The identification of sequence innovations in the genomes of mammals facilitates understanding of human gene function, as well as sheds light on the molecular mechanisms which underlie these changes. Although gene duplication plays a major role in genome evolution, studies regarding concerted evolution events among gene family members have been limited in scope and restricted to protein-coding regions, where high sequence similarity is easily detectable. RESULTS: We describe a mammalian-specific expansion of more than 20 rapidly evolving genes on human chromosome Xq22.1. Many of these are highly divergent in their protein-coding regions yet contain a conserved sequence motif in their 5' UTRs which appears to have been maintained by multiple events of concerted evolution. These events have led to the generation of chimaeric genes, each with a 5' UTR and a protein-coding region that possess independent evolutionary histories. We suggest that concerted evolution has occurred via gene conversion independently in different mammalian lineages, and these events have resulted in elevated G+C levels in the encompassing genomic regions. These concerted evolution events occurred within and between genes from three separate protein families ('brain-expressed X-linked' [BEX], WWbp5-like X-linked [WEX] and G protein-coupled receptor-associated sorting protein [GASP]), which often are expressed in mammalian brains and associated with receptor mediated signalling and apoptosis. CONCLUSION: Despite high protein-coding divergence among mammalian specific genes, we identified a DNA motif common to these genes' 5' UTR exons. The motif has undergone concerted evolution events independently of its neighbouring protein-coding regions, leading to formation of evolutionary chimaeric genes. These findings have implications for the identification of non protein-coding regulatory elements and their lineage-specific evolution in mammals. PMID- 16221303 TI - Prosthetic valve endocarditis caused by Pseudomonas luteola. AB - BACKGROUND: Pseudomonas luteola has been recognized as an uncommon cause of bacteremia and of infections in patients with underlying medical disorders. CASE PRESENTATION: We isolated P. luteola from blood cultures in a patient with prosthetic valve endocarditis developed 16 months after cardiac surgery. CONCLUSION: P. luteola is a rare opportunistic agent, with a propensity of infecting valvular prostheses. PMID- 16221304 TI - Phydbac "Gene Function Predictor": a gene annotation tool based on genomic context analysis. AB - BACKGROUND: The large amount of completely sequenced genomes allows genomic context analysis to predict reliable functional associations between prokaryotic proteins. Major methods rely on the fact that genes encoding physically interacting partners or members of shared metabolic pathways tend to be proximate on the genome, to evolve in a correlated manner and to be fused as a single sequence in another organism. RESULTS: The new "Gene Function Predictor", linked to the web server Phydbac proposes putative associations between Escherichia coli K-12 proteins derived from a combination of these methods. We show that associations made by this tool are more accurate than linkages found in the other established databases. Predicted assignments to GO categories, based on pre existing functional annotations of associated proteins are also available. This new database currently holds 9,379 pairwise links at an expected success rate of at least 80%, the 6,466 functional predictions to GO terms derived from these links having a level of accuracy higher than 70%. CONCLUSION: The "Gene Function Predictor" is an automatic tool that aims to help biologists by providing them hypothetical functional predictions out of genomic context characteristics. The "Gene Function predictor" is available at http://www.igs.cnrs mrs.fr/phydbac/indexPS.html. PMID- 16221305 TI - Nonnatural amino acid incorporation into the methionine 214 position of the metzincin Pseudomonas aeruginosa alkaline protease. AB - BACKGROUND: The alkaline protease from Pseudomonas aeruginosa (AprA) is a member of the metzincin superfamily of metalloendoproteases. A key feature of these proteases is a conserved methionine-containing 1,4-tight beta turn at the base of the active site zinc binding region. RESULTS: To explore the invariant methionine position in this class of protease, incorporation of a nonnatural fluorinated methionine, L-difluoromethionine (DFM), into this site was accomplished. Although overproduction of the N-terminal catalytic fragment of AprA resulted in protein aggregates which could not be resolved, successful heterologous production of the entire AprA was accomplished in the presence and absence of the nonnatural amino acid. DFM incorporation was found to only slightly alter the enzyme kinetics of AprA. In addition, differential scanning calorimetry indicated no significant alteration in the thermal stability of the modified enzyme. CONCLUSION: Although invariant in all metzincin proteases, the methionine 214 position in AprA can be successfully replaced by the nonnatural amino acid DFM resulting in little effect on protein structure and function. This study indicates that the increased size of the methyl group by the introduction of two fluorines is still sufficiently non-sterically demanding, and bodes well for the application of DFM to biophysical studies of protein structure and function in this class of protease. PMID- 16221306 TI - Methods for confidence interval estimation of a ratio parameter with application to location quotients. AB - BACKGROUND: The location quotient (LQ) ratio, a measure designed to quantify and benchmark the degree of relative concentration of an activity in the analysis of area localization, has received considerable attention in the geographic and economics literature. This index can also naturally be applied in the context of population health to quantify and compare health outcomes across spatial domains. However, one commonly observed limitation of LQ is its widespread use as only a point estimate without an accompanying confidence interval. METHODS: In this paper we present statistical methods that can be used to construct confidence intervals for location quotients. The delta and Fieller's methods are generic approaches for a ratio parameter and the generalized linear modelling framework is a useful re-parameterization particularly helpful for generating profile likelihood based confidence intervals for the location quotient. A simulation experiment is carried out to assess the performance of each of the analytic approaches and a health utilization data set is used for illustration. RESULTS: Both the simulation results as well as the findings from the empirical data show that the different analytical methods produce very similar confidence limits for location quotients. When incidence of outcome is not rare and sample sizes are large, the confidence limits are almost indistinguishable. The confidence limits from the generalized linear model approach might be preferable in small sample situations. CONCLUSION: LQ is a useful measure which allows quantification and comparison of health and other outcomes across defined geographical regions. It is a very simple index to compute and has a straightforward interpretation. Reporting this estimate with appropriate confidence limits using methods presented in this paper will make the measure particularly attractive for policy and decision makers. PMID- 16221308 TI - An adaptive prediction and detection algorithm for multistream syndromic surveillance. AB - BACKGROUND: Surveillance of Over-the-Counter pharmaceutical (OTC) sales as a potential early indicator of developing public health conditions, in particular in cases of interest to biosurvellance, has been suggested in the literature. This paper is a continuation of a previous study in which we formulated the problem of estimating clinical data from OTC sales in terms of optimal LMS linear and Finite Impulse Response (FIR) filters. In this paper we extend our results to predict clinical data multiple steps ahead using OTC sales as well as the clinical data itself. METHODS: The OTC data are grouped into a few categories and we predict the clinical data using a multichannel filter that encompasses all the past OTC categories as well as the past clinical data itself. The prediction is performed using FIR (Finite Impulse Response) filters and the recursive least squares method in order to adapt rapidly to nonstationary behaviour. In addition, we inject simulated events in both clinical and OTC data streams to evaluate the predictions by computing the Receiver Operating Characteristic curves of a threshold detector based on predicted outputs. RESULTS: We present all prediction results showing the effectiveness of the combined filtering operation. In addition, we compute and present the performance of a detector using the prediction output. CONCLUSION: Multichannel adaptive FIR least squares filtering provides a viable method of predicting public health conditions, as represented by clinical data, from OTC sales, and/or the clinical data. The potential value to a biosurveillance system cannot, however, be determined without studying this approach in the presence of transient events (nonstationary events of relatively short duration and fast rise times). Our simulated events superimposed on actual OTC and clinical data allow us to provide an upper bound on that potential value under some restricted conditions. Based on our ROC curves we argue that a biosurveillance system can provide early warning of an impending clinical event using ancillary data streams (such as OTC) with established correlations with the clinical data, and a prediction method that can react to nonstationary events sufficiently fast. Whether OTC (or other data streams yet to be identified) provide the best source of predicting clinical data is still an open question. We present a framework and an example to show how to measure the effectiveness of predictions, and compute an upper bound on this performance for the Recursive Least Squares method when the following two conditions are met: (1) an event of sufficient strength exists in both data streams, without distortion, and (2) it occurs in the OTC (or other ancillary streams) earlier than in the clinical data. PMID- 16221307 TI - Serum hyaluronate as a non-invasive marker of hepatic fibrosis and inflammation in HBeAg-negative chronic hepatitis B. AB - BACKGROUND: HBV infection is a serious global heath problem. It is crucial to monitor this disease more closely with a non-invasive marker in clinical trials. We aimed to evaluate the predictive value of serum hyaluronate for the presence of extensive liver fibrosis and inflammation. METHODS: 28 healthy volunteers and 65 patients with HBeAg negative chronic hepatitis B were enrolled. Liver biopsies scored according to Ishak system. Association of serum hyaloronate with liver fibrosis and inflammation were assessed, and cut off points for serum hyaluronate levels were identified by receiver operating characteristics (ROC) curves and their values for prediction of fibrosis and inflammation were assessed. RESULTS: In patients with CHB serum hyaluronate had the most significant correlation and predictive values for the liver fibrosis and inflammation comparing to the other variables. At the cut off point of 126.4 ngm/ml it could discriminate extensive fibrosis from milder ones with sensitivity of 90.9% and specificity of 98.1%. With the same value it could discriminate extensive inflammation from their milder counterparts with sensitivity of 63.6% and specificity of 92.6%. CONCLUSION: Serum hyaluronate was the best predictor of extensive liver fibrosis and inflammation and it could discriminate subgroups of patients with chronic hepatitis B. It could be used as a non-invasive test to monitor these patients more closely with developing anti viral agents in clinical trials. PMID- 16221309 TI - Better prediction of protein contact number using a support vector regression analysis of amino acid sequence. AB - BACKGROUND: Protein tertiary structure can be partly characterized via each amino acid's contact number measuring how residues are spatially arranged. The contact number of a residue in a folded protein is a measure of its exposure to the local environment, and is defined as the number of Cbeta atoms in other residues within a sphere around the Cbeta atom of the residue of interest. Contact number is partly conserved between protein folds and thus is useful for protein fold and structure prediction. In turn, each residue's contact number can be partially predicted from primary amino acid sequence, assisting tertiary fold analysis from sequence data. In this study, we provide a more accurate contact number prediction method from protein primary sequence. RESULTS: We predict contact number from protein sequence using a novel support vector regression algorithm. Using protein local sequences with multiple sequence alignments (PSI-BLAST profiles), we demonstrate a correlation coefficient between predicted and observed contact numbers of 0.70, which outperforms previously achieved accuracies. Including additional information about sequence weight and amino acid composition further improves prediction accuracies significantly with the correlation coefficient reaching 0.73. If residues are classified as being either "contacted" or "non-contacted", the prediction accuracies are all greater than 77%, regardless of the choice of classification thresholds. CONCLUSION: The successful application of support vector regression to the prediction of protein contact number reported here, together with previous applications of this approach to the prediction of protein accessible surface area and B-factor profile, suggests that a support vector regression approach may be very useful for determining the structure-function relation between primary protein sequence and higher order consecutive protein structural and functional properties. PMID- 16221310 TI - SPdb--a signal peptide database. AB - BACKGROUND: The signal peptide plays an important role in protein targeting and protein translocation in both prokaryotic and eukaryotic cells. This transient, short peptide sequence functions like a postal address on an envelope by targeting proteins for secretion or for transfer to specific organelles for further processing. Understanding how signal peptides function is crucial in predicting where proteins are translocated. To support this understanding, we present SPdb signal peptide database http://proline.bic.nus.edu.sg/spdb, a repository of experimentally determined and computationally predicted signal peptides. RESULTS: SPdb integrates information from two sources (a) Swiss-Prot protein sequence database which is now part of UniProt and (b) EMBL nucleotide sequence database. The database update is semi-automated with human checking and verification of the data to ensure the correctness of the data stored. The latest release SPdb release 3.2 contains 18,146 entries of which 2,584 entries are experimentally verified signal sequences; the remaining 15,562 entries are either signal sequences that fail to meet our filtering criteria or entries that contain unverified signal sequences. CONCLUSION: SPdb is a manually curated database constructed to support the understanding and analysis of signal peptides. SPdb tracks the major updates of the two underlying primary databases thereby ensuring that its information remains up-to-date. PMID- 16221311 TI - The McKenzie Method for the management of acute non-specific low back pain: design of a randomised controlled trial [ACTRN012605000032651]. AB - BACKGROUND: Low back pain (LBP) is a major health problem. Effective treatment of acute LBP is important because it prevents patients from developing chronic LBP, the stage of LBP that requires costly and more complex treatment. Physiotherapists commonly use a system of diagnosis and exercise prescription called the McKenzie Method to manage patients with LBP. However, there is insufficient evidence to support the use of the McKenzie Method for these patients. We have designed a randomised controlled trial to evaluate whether the addition of the McKenzie Method to general practitioner care results in better outcomes than general practitioner care alone for patients with acute LBP. METHODS/DESIGN: This paper describes the protocol for a trial examining the effects of the McKenzie Method in the treatment of acute non-specific LBP. One hundred and forty eight participants who present to general medical practitioners with a new episode of acute non-specific LBP will be randomised to receive general practitioner care or general practitioner care plus a program of care based on the McKenzie Method. The primary outcomes are average pain during week 1, pain at week 1 and 3 and global perceived effect at week 3. DISCUSSION: This trial will provide the first rigorous test of the effectiveness of the McKenzie Method for acute non-specific LBP. PMID- 16221312 TI - Efficient expression of transgenes in adult zebrafish by electroporation. AB - BACKGROUND: Expression of transgenes in muscle by injection of naked DNA is widely practiced. Application of electrical pulses at the site of injection was demonstrated to improve transgene expression in muscle tissue. Zebrafish is a precious model to investigate developmental biology in vertebrates. In this study we investigated the effect of electroporation on expression of transgenes in 3-6 month old adult zebrafish. RESULTS: Electroporation parameters such as number of pulses, voltage and amount of plasmid DNA were optimized and it was found that 6 pulses of 40 V.cm(-1) at 15 microg of plasmid DNA per fish increased the luciferase expression 10-fold compared to controls. Similar enhancement in transgene expression was also observed in Indian carp (Labeo rohita). To establish the utility of adult zebrafish as a system for transient transfections, the strength of the promoters was compared in A2 cells and adult zebrafish after electroporation. The relative strengths of the promoters were found to be similar in cell lines and in adult zebrafish. GFP fluorescence in tissues after electroporation was also studied by fluorescence microscopy. CONCLUSION: Electroporation after DNA injection enhances gene expression 10-fold in adult zebrafish. Electroporation parameters for optimum transfection of adult zebrafish with tweezer type electrode were presented. Enhanced reporter gene expression upon electroporation allowed comparison of strengths of the promoters in vivo in zebrafish. PMID- 16221313 TI - The epidemiology and modern management of traumatic hemorrhage: US and international perspectives. AB - Trauma is a worldwide problem, with severe and wide ranging consequences for individuals and society as a whole. Hemorrhage is a major contributor to the dilemma of traumatic injury and its care. In this article we describe the international epidemiology of traumatic injury, its causes and its consequences, and closely examine the role played by hemorrhage in producing traumatic morbidity and mortality. Emphasis is placed on defining situations in which traditional methods of hemorrhage control often fail. We then outline and discuss modern principles in the management of traumatic hemorrhage and explore developing changes in these areas. We conclude with a discussion of outcome measures for the injured patient within the context of the epidemiology of traumatic injury. PMID- 16221314 TI - Optimizing the use of blood products in trauma care. AB - Blood transfusion has been used to treat the injured since the US Civil War. Now, it saves the lives of tens of thousands of injured patients each year. However, not everyone who receives blood benefits, and some recipients are injured by the transfusion itself. Effective blood therapy in trauma management requires an integration of information from diverse sources, including data relating to trauma and blood use epidemiology, medical systems management, and clinical care. Issues of current clinical concern in highly developed trauma systems include how to manage massive transfusion events, how to limit blood use and so minimize exposure to transfusion risks, how to integrate new hemorrhage control modalities, and how to deal with blood shortages. Less developed trauma systems are primarily concerned with speeding transport to specialized facilities and assembling trauma center resources. This article reviews the factors that effect blood use in urgent trauma care. PMID- 16221315 TI - Mechanistic implications for the use and monitoring of recombinant activated factor VII in trauma. AB - As interest in the use of activated recombinant factor VII (rFVIIa) in trauma grows, questions arise regarding how best to monitor rFVIIa therapy and when rFVIIa may be expected to improve hemostasis. Knowledge of the mechanisms of action may be combined with available data on laboratory monitoring and efficacy in various coagulopathic states in coming to clinically relevant conclusions. This review addresses the physiology of hemostasis, placing emphasis on how rFVIIa influences the process by both tissue factor dependent and tissue factor independent mechanisms. This is extended to a mechanistic consideration of how rFVIIa may function under acidotic, hypothermic, and hemodilutional and/or consumptive conditions of trauma related coagulopathy. When these considerations are viewed alongside the available clinical data, it becomes apparent that rFVIIa has potential to improve hemostasis during trauma coagulopathy, within limitations. Common laboratory procedures are discussed with reference to mechanisms of action of rFVIIa and the available clinical data. Although there is no single assay that can predict rFVIIa efficacy in trauma, the prothrombin time (PT) is recommended as a minimum. Although a shortened PT does not predict success, correction of PT into the normal range may be a better indicator. A nonresponding PT appears to indicate that rFVIIa alone will not lead to hemostasis, and that additional blood products and other measures must be applied. Once the patient is more stable, PT and thromboelastography are recommended. PMID- 16221316 TI - Preclinical trauma studies of recombinant factor VIIa. AB - Preclinical studies in animals and ex vivo human blood have provided a solid rationale for conducting prospective randomized trials in trauma patients. Small animal models have been utilized to study the efficacy of recombinant activated factor VII (rFVIIa; NovoSeven) in treating thrombocytopenic rabbits and for the reversal of anticoagulation. Safety models in the rabbit also exist to test for systemic activation of clotting and pathologic thrombosis. Animal models simulating traumatic injuries in humans have primarily been performed in pigs because of species similarities in terms of coagulation characteristics and the larger internal organs. The pig studies, utilizing human rFVIIa, have shown increased strength of clot formation, decreased bleeding, and improved survival. However, these findings are not uniform and are dependant on the model chosen. All of the animal models described have provided good safety data and suggest that the use of rFVIIa is not associated with systemic activation of coagulation or microthrombosis of end organs. PMID- 16221319 TI - Professions, generations and reproductive dynamics of a French Alpine population (16th-20th centuries). AB - As part of a survey of the biological history of Alpine populations, the lineages of all the families of the Vallouise valley (a French 'department' of the Hautes Alpes) have been reconstructed over several centuries. The genealogies have been included in a computerized population record, known as 'Vallouise in the Briancon area (14th-20th centuries)', using the French-Canadian programme Analypop. Most of the professions of the family heads were included in the files. In this study, various profession groups were identified and their descents determined over successive generations. In this mountain area, where over 92% of marriages took place among relatives during the 19th century, the profession groups modulated their descents according to chosen strategies, sometimes with considerable differences among groups but with a remarkable consistency of behaviour. Moreover, there was weak interpenetration in the descents of each profession at both the 2nd and 3rd generations. PMID- 16221317 TI - Clinical experiences and current evidence for therapeutic recombinant factor VIIa treatment in nontrauma settings. AB - The hemostatic properties of recombinant activated factor VII (rFVIIa) are established in patients with inherited or acquired hemophilia with inhibitors and in patients with congenital factor VII deficiencies. Emerging clinical evidence suggests that there may be a wider role for rFVIIa in the management of hemorrhage associated with traumatic injury/accident and severe bleeding associated with critical surgery. This article considers recent data from studies in which rFVIIa was used in an attempt to control bleeding in clinical situations as diverse as coagulopathy associated with chronic liver disease, massive perioperative bleeding and bleeding during prostatectomy, organ transplantation and orthopedic surgery, uncontrollable obstetric hemorrhage, and intracerebral hemorrhage. In nontrauma settings involving acute and potentially life threatening bleeding, there may be a place for rFVIIa as adjunctive therapy in the control of hemostasis. PMID- 16221318 TI - Recombinant activated factor VIIa and hemostasis in critical care: a focus on trauma. AB - In this article we describe the current use of recombinant activated factor VII (rFVIIa; NovoSeven) in trauma patients. Emphasis is placed on current uses as defined by key studies, efficacy data, and safety data. Most published studies in trauma patients are retrospective case studies and reports, although an international, double-blind, randomized, controlled, phase II study has been conducted that reported on the efficacy of rFVIIa in reducing the amount of blood products transfused in blunt trauma patients. That study demonstrated the efficacy and safety profile of this hemostatic agent as compared with placebo as adjunctive therapy in the management of severe bleeding associated with trauma. Further prospective, randomized, and placebo-controlled clinical trials will yield more information on the role of rFVIIa in the management of traumatic bleeding. PMID- 16221320 TI - Determinants of spacing contraceptive use among couples in Mumbai: a male perspective. AB - This study aimed to determine the factors influencing the use of spacing contraceptive methods in India, particularly from men's perspective. Data were obtained through a semi-structured interview schedule from 2,687 married men aged between 18 and 40 years from central Mumbai City, India, during 1999. Chi-squared tests and binary logistic regression analysis was carried out to determine the relationship between various variables and the likelihood of a couple using spacing contraceptive methods. Of the 2,687 couples, 1,395 (51.9%) were using one or other method of spacing contraceptives and 1,292 (48.1%) were not using any method at the time of survey. Male participation in contraceptive use was 23% (condom and withdrawal). The results indicate that the use of spacing contraceptive methods was significantly higher among those couples where the men desired one or two children (OR=4.3), had knowledge of five or more contraceptive methods (OR=1.9) and discussed with their wives obtaining family planning information (OR=3.2), spacing (OR=2.7) and permanent (OR=2) contraceptive methods. Age, income, desired number of children, knowledge of a greater number of contraceptive methods, inter-spouse communication regarding obtaining family planning information, spacing and permanent methods were found to be strong predictors of the use of spacing contraceptive methods. The study underlines the importance of intervention programmes aimed at promoting a small family norm, increasing the number of contraceptive choices available and encouraging inter spouse communication. Hence, policymakers and programme managers should encourage interventions in this direction, targeting couples to enhance the use of spacing contraceptive methods. PMID- 16221321 TI - Insurance and other socioeconomic determinants of elderly longevity in a Costa Rican panel. AB - Official figures show that life expectancy in Costa Rica is longer than in the United States (US), in spite of the fact that per capita health expenditure is only one-tenth that of the US. To check whether this is for real and to explore some of its determinants, 900 Costa Ricans aged 60+ were followed from 1984 to 2001. Follow-up household visits were made, deaths were tracked in the national death registry, and survival status in the voting registry was double-checked. In addition, the survivors were contacted in 2002. Two-thirds of the panel had died by December 2001. Kaplan-Meier curves, life tables and Cox regression were used to analyse the panel's survival. Mortality in the panel was slightly higher than the Costa Rican average and similar to that in the US, confirming the exceptional longevity of Costa Ricans. Survival was substantially lower among unmarried men and individuals with limited autonomy at the beginning of the study. The effect of socioeconomic status is weak. Insurance effects seem to be confounded by selection biases. PMID- 16221322 TI - National intelligence, suicide rate in the elderly, and a threshold intelligence for suicidality: an ecological study of 48 Eurasian countries. AB - Across 85 countries around the world, Voracek (2004) found a significant positive relation between estimated national intelligence (IQ) and national male and female suicide rate. The relation was not attenuated when countries' per capita Gross Domestic Product (GDP) and type of national IQ estimation were statistically controlled. Independently, investigating the total national suicide rate only, Lester (2003) arrived at the same conclusion. These two findings are consistent with a corollary of de Catanzaro's (1981) evolutionary theory of human suicide, namely that a threshold intelligence is necessary for suicidality and that intelligence and suicide mortality should thus be positively related. Here, further evidence for this hypothesis is bolstered by focusing on suicide rates of the elderly. Across 48 Eurasian countries, estimated national IQ was significantly positively related to national suicide rates of people aged 65 years and over. This new ecological-level finding survived statistical controlling for a set of seven variables (type of national IQ estimation, national GDP, stableness and recency measures for suicide rates, and rates of adult literacy, urbanization and Roman Catholics), which thus are not confounding factors for the relation of intelligence and suicide mortality. Based on ecological data, the threshold IQ for suicidality is predicted to be 70 or slightly over, an estimate that is consistent with various suicidological observations. PMID- 16221323 TI - Using indirect methods to understand the impact of forced migration on long-term under-five mortality. AB - Despite the large numbers of displaced persons and the often-lengthy periods of displacement, little is known about the impact of forced migration on long-term under-five mortality. This paper looks at the Brass Method (and adaptations of this method) and the Preceding Birth Technique in combination with a classification of women by their migration and reproductive histories, in order to study the impact of forced migration on under-five mortality. Data came from the Demography of Forced Migration Project, a study on mortality, fertility and violence in the refugee and host populations of Arua District, Uganda and Yei River District, Sudan. Results indicate that women who did not migrate in a situation of conflict and women who repatriated before the age of 15, had children with the highest under-five mortality rates compared with women who were currently refugees and women who repatriated after the age of 15. PMID- 16221324 TI - Self-reported malaria and mosquito avoidance in relation to household risk factors in a Kenyan coastal city. AB - A geographically stratified cross-sectional survey was conducted in 2002 to investigate household-level factors associated with use of mosquito control measures and self-reported malaria in Malindi, Kenya. A total of 629 households were surveyed. Logistic regressions were used to analyse the data. Half of all households (51%) reported all occupants using an insecticide-treated bed net and at least one additional mosquito control measure such as insecticides or removal of standing water. Forty-nine per cent reported a history of malaria in the household. Of the thirteen household factors analysed, low (OR=0.23, CI 0.11, 0.48) and medium (OR=0.50, CI 0.29, 0.86) education, mud--wood--coral (OR=0.0.39, CI 0.24, 0.66) and mud block--plaster (OR=0.47, CI 0.25, 0.87) wall types, farming (OR=1.38, CI 1.01, 1.90) and travel to rural areas (OR=0.48, CI 0.26, 0.91) were significantly associated with the use of mosquito control, while controlling for other covariates in the model. History of reported malaria was not associated with the use of mosquito control (OR=1.22, CI 0.79, 1.88). Of the thirteen covariates analysed in the second model, only two household factors were associated with history of malaria: being located in the well-drained stratum (OR=0.49, CI 0.26, 0.96) and being bitten while in the house (OR=1.22, CI 0.19, 0.49). These results suggest that high socioeconomic status is associated with increased household-level mosquito control use, although household-level control may not be enough, as many people are exposed to biting mosquitoes while away from the house and in areas that are more likely to harbour mosquitoes. PMID- 16221325 TI - Socioeconomic status does not moderate the familiality of cognitive abilities in the Hawaii family study of cognition. AB - Data from 949 families of Caucasian and 400 families of Japanese ancestry who took part in the Hawaii Family Study of Cognition were used to ascertain the associations of parental cognitive ability, parental education and paternal occupation with offspring cognitive ability. In particular, analyses were focused on testing the possible moderating effects of parental socioeconomic status on the familial transmission of cognitive abilities. Parental cognitive ability was substantially associated and parental education and paternal occupation only trivially associated with offspring performance. In contrast to the findings of Turkheimer et al. (2003), there was no evidence in these data that familiality for cognitive abilities was lower in the lower as opposed to upper levels of socioeconomic status. These results were consistent across measures, ethnicity and sex of offspring. PMID- 16221327 TI - The Paleogene fossil record of birds in Europe. AB - The Paleogene (Paleocene-Oligocene) fossil record of birds in Europe is reviewed and recent and fossil taxa are placed into a phylogenetic framework, based on published cladistic analyses. The pre-Oligocene European avifauna is characterized by the complete absence of passeriform birds, which today are the most diverse and abundant avian taxon. Representatives of small non-passeriform perching birds thus probably had similar ecological niches before the Oligocene to those filled by modern passerines. The occurrence of passerines towards the Lower Oligocene appears to have had a major impact on these birds, and the surviving crown-group members of many small arboreal Eocene taxa show highly specialized feeding strategies not found or rare in passeriform birds. It is detailed that no crown-group members of modern 'families' are known from pre Oligocene deposits of Europe, or anywhere else. The phylogenetic position of Paleogene birds thus indicates that diversification of the crown-groups of modern avian 'families' did not take place before the Oligocene, irrespective of their relative position within Neornithes (crown-group birds). The Paleogene fossil record of birds does not even support crown-group diversification of Galliformes, one of the most basal taxa of neognathous birds, before the Oligocene, and recent molecular studies that dated diversification of galliform crown-group taxa into the Middle Cretaceous are shown to be based on an incorrect interpretation of the fossil taxa used for molecular clock calibrations. Several taxa that occur in the Paleogene of Europe have a very different distribution than their closest extant relatives. The modern survivors of these Paleogene lineages are not evenly distributed over the continents, and especially the great number of taxa that are today restricted to South and Central America is noteworthy. The occurrence of stem-lineage representatives of many taxa that today have a restricted Southern Hemisphere distribution conflicts with recent hypotheses on a Cretaceous vicariant origin of these taxa, which were deduced from the geographical distribution of the basal crown-group members. PMID- 16221328 TI - Molecular and morphological supertree of stony corals (Anthozoa: Scleractinia) using matrix representation parsimony. AB - The supertree algorithm matrix representation with parsimony was used to combine existing hypotheses of coral relationships and provide the most comprehensive species-level estimate of scleractinian phylogeny, comprised of 353 species (27% of extant species), 141 genera (63%) and 23 families (92%) from all seven suborders. The resulting supertree offers a guide for future studies in coral systematics by highlighting regions of concordance and conflict in existing source phylogenies. It should also prove useful in formal comparative studies of character evolution. Phylogenetic effort within Scleractinia has been taxonomically uneven, with a third of studies focussing on the Acroporidae or its most diverse genera. Sampling has also been geographically non-uniform, as tropical, reef-forming taxa have been considered twice as often as non-reef species. The supertree indicated that source trees concur on numerous aspects of coral relationships, such as the division between robust versus complex corals and the distant relationship between families in Archaeocoeniina. The supertree also supported the existence of a large, taxonomically diverse and monophyletic group of corals with many Atlantic representatives having exsert corallites. Another large, unanticipated clade consisted entirely of solitary deep-water species from three families. Important areas of ambiguity include the relationship of Astrocoeniidae to Pocilloporidae and the relative positions of several, mostly deep-water genera of Caryophylliidae. Conservative grafting of species at the base of congeneric groups with uncontroversial monophyletic status resulted in a more comprehensive, though less resolved tree of 1016 taxa. PMID- 16221329 TI - How is female mate choice affected by male competition? AB - The plethora of studies devoted to the topics of male competition and female mate choice belie the fact that their interaction remains poorly understood. Indeed, on the question of whether competition should help or hinder the choice process, opinions scattered throughout the sexual selection literature seem unnecessarily polarised. We argue, in the light of recent theoretical and empirical advances, that the effect of competition on mate choice depends on whether it results in the choosy sex attaining high breeding value for total fitness, considering both direct and indirect fitness benefits. Specifically, trade-offs may occur between different fitness benefits if some are correlated with male competitive ability whilst others are not. Moreover, the costs and benefits of mating with competitive males may vary in time and/or space. These considerations highlight the importance of injecting a life-history perspective into sexual selection studies. Within this context, we turn to the sexual selection literature to try to offer insights into the circumstances when competition might be expected to have positive or negative implications for pre-copulatory female choice. In this regard, we elaborate on three stages where competition might impact upon the choice process: (i) during mate detection, (ii) mate evaluation, and (iii) in dictating actual mating outcomes. We conclude by offering researchers several potentially rewarding avenues for future research. PMID- 16221330 TI - The role of eyespots as anti-predator mechanisms, principally demonstrated in the Lepidoptera. AB - Eyespots are found in a variety of animals, in particular lepidopterans. The role of eyespots as antipredator mechanisms has been discussed since the 19th Century, with two main hypotheses invoked to explain their occurrence. The first is that large, centrally located eyespots intimidate predators by resembling the eyes of the predators' own enemies; the second, though not necessarily conflicting, hypothesis is that small, peripherally located eyespots function as markers to deflect the attacks of predators to non-vital regions of the body. A third possibility is also proposed; that eyespots intimidate predators merely because they are novel or rarely encountered salient features. These hypotheses are reviewed, with special reference given to avian predators, since these are likely to be the principal visually hunting predators of the lepidopterans considered. Also highlighted is the necessity to consider the potential influence of sexual selection on lepidopteran wing patterns, and the genetics and development of eyespot formation. PMID- 16221331 TI - Environments and evolution: interactions between stress, resource inadequacy and energetic efficiency. AB - Evolutionary change is interpreted in terms of the near-universal ecological scenario of stressful environments. Consequently, there is a premium on the energetically efficient exploitation of resources in a resource-inadequate world. Under this environmental model, fitness can be approximated to energetic efficiency especially towards the limits of survival. Furthermore, fitness at one stage of the life-cycle should correlate with fitness at other stages, especially for development time, survival and longevity; 'good genotypes' under stress should therefore be at a premium. Conservation in the wild depends primarily on adaptation to abiotically changing habitats since towards the limits of survival, genomic variation is rarely restrictive. The balance between energetic costs under variable environments and energy from resources provides a model for interpreting evolutionary stasis, punctuational and gradual change, and specialist diversification. Ultimately, a species should be in an equilibrium between the physiology of an organism and its adaptation to the environment. The primary key to understanding evolutionary change should therefore be ecological, highlighting energy availability in a stressed world; this approach is predictive for various patterns of evolutionary change in the living and fossil biota. PMID- 16221332 TI - Beyond the '3/4-power law': variation in the intra- and interspecific scaling of metabolic rate in animals. AB - In this review I show that the '3/4-power scaling law' of metabolic rate is not universal, either within or among animal species. Significant variation in the scaling of metabolic rate with body mass is described mainly for animals, but also for unicells and plants. Much of this variation, which can be related to taxonomic, physiological, and/or environmental differences, is not adequately explained by existing theoretical models, which are also reviewed. As a result, synthetic explanatory schemes based on multiple boundary constraints and on the scaling of multiple energy-using processes are advocated. It is also stressed that a complete understanding of metabolic scaling will require the identification of both proximate (functional) and ultimate (evolutionary) causes. Four major types of intraspecific metabolic scaling with body mass are recognized [based on the power function R=aMb, where R is respiration (metabolic) rate, a is a constant, M is body mass, and b is the scaling exponent]: Type I: linear, negatively allometric (b<1); Type II: linear, isometric (b=1); Type III: nonlinear, ontogenetic shift from isometric (b=1), or nearly isometric, to negatively allometric (b<1); and Type IV: nonlinear, ontogenetic shift from positively allometric (b>1) to one or two later phases of negative allometry (b<1). Ontogenetic changes in the metabolic intensity of four component processes (i.e. growth, reproduction, locomotion, and heat production) appear to be important in these different patterns of metabolic scaling. These changes may, in turn, be shaped by age (size)-specific patterns of mortality. In addition, major differences in interspecific metabolic scaling are described, especially with respect to mode of temperature regulation, body-size range, and activity level. A 'metabolic-level boundaries hypothesis' focusing on two major constraints (surface-area limits on resource/waste exchange processes and mass/volume limits on power production) can explain much, but not all of this variation. My analysis indicates that further empirical and theoretical work is needed to understand fully the physiological and ecological bases for the considerable variation in metabolic scaling that is observed both within and among species. Recommended approaches for doing this are discussed. I conclude that the scaling of metabolism is not the simple result of a physical law, but rather appears to be the more complex result of diverse adaptations evolved in the context of both physico-chemical and ecological constraints. PMID- 16221333 TI - Evolutionary ecology of facultative paedomorphosis in newts and salamanders. AB - Facultative paedomorphosis is an environmentally induced polymorphism that results in the coexistence of mature, gilled, and fully aquatic paedomorphic adults and transformed, terrestrial, metamorphic adults in the same population. This polymorphism has been of interest to scientists for decades because it occurs in a large number of caudate amphibian taxa as well as in a large diversity of habitats. Numerous experimental and observational studies have been conducted to explain the proximate and ultimate factors affecting these heterochronic variants in natural populations. The production of each alternative phenotype is based on a genotypexenvironment interaction and research suggests that differences in the environment can produce paedomorphs through several ontogenetic pathways. No single advantage accounts for the maintenance of this polymorphism. Rather, the interplay of different costs and benefits explains the success of the polyphenism across variable environments. Facultative paedomorphosis allows individuals to cope with habitat variation, to take advantage of environmental heterogeneity in the presence of open niches, and to increase their fitness. This process is expected to constitute a first step towards speciation events, and is also an example of biodiversity at the intraspecific level. The facultative paedomorphosis system is thus ripe for future studies encompassing ecology, evolution, behaviour, endocrinology, physiology, and conservation biology. Few other systems have been broad enough to provide varied research opportunities on topics as diverse as phenotypic plasticity, speciation, mating behaviour, and hormonal regulation of morphology. Further research on facultative paedomorphosis will provide needed insight into these and other important questions facing biologists. PMID- 16221335 TI - [Re-consideration of certain issues in diagnosis and treatment of glomerular diseases in children]. PMID- 16221336 TI - [Analysis of 1268 patients with chronic renal failure in childhood: a report from 91 hospitals in China from 1990 to 2002]. AB - OBJECTIVE: Chronic renal failure (CRF) of childhood is not rare. The prognosis of CRF is very poor because of severe systemic complications. A nation-wide survey was conducted and data of hospitalized children (younger than 14 years old) with CRF during the period of 1990 to 2002 were analyzed. The aim was to investigate the epidemiology, natural history, clinical-pathological characteristics, treatment and outcome of the hospitalized children with CRF. METHODS: Questionnaires concerning children with CRF were designed and distributed to the doctors of 91 hospitals in China. The criterion of CRF was creatinine clearance (CCr) < 50 ml/(min x 1.73 m(2)). The data were collected and analyzed. RESULTS: From January 1, 1990 to December 31, 2002, 1658 hospitalized children were diagnosed as CRF. The average annual cases of childhood CRF accounted for 1.31% (ranged from 0.72% to 1.75%) of the hospitalized cases with urologic-kidney diseases. In a comparison between 1990 - 1996 and 1997 - 2002, there were significant increases in the average annual number of cases of childhood CRF and the case ratio of CRF to urologic-kidney diseases (82 +/- 27 vs. 181 +/- 45 and 0.98 +/- 0.21 vs. 1.56 +/- 0.17, respectively, P < 0.001). Complete records were available for 1268 patients. The male to female ratio was 1.49:1. The mean age at the disease onset was 8.18 years. The mean duration of pre-diagnosis of CRF was 2.53 years. In this study, the main primary renal diseases causing CRF were chronic glomerulonephritis and nephrotic syndrome (52.7%). One-fourth of all cases had congenital and hereditary renal diseases, and the majority were renal hypoplasia and dysplasia. The main manifestations of CRF were anemia, gastrointestinal disorders, edema, hypertension and growth retardation. The mean serum creatinine and BUN were 594.7 micromol/L and 39.1 mmol/L, respectively. The cases with renal function >or= grade IV accounted for 80% of all cases. By renal ultrasound scanning, one-third of CRF children were found to have renal atrophy and a part of patients had cystic disorder. Most of the cases received conservative treatment. Dialysis therapy (including 66.5% of hemodialysis and 33.5% of peritoneal) was given to 15.8% of the patients. Twenty-nine cases received renal transplantation. The rate of graft survival was 93.1%. Follow-up was carried out for to 230 cases, the mean duration of follow-up was 2.36 years. One hundred and sixty-seven patients died during hospitalization over the 13-year review period. The main causes of death were cardiac failure and infections in addition to uremia. CONCLUSION: The incidence of CRF in children showed an increasing trend year after year. The main age of onset of the disease was school age. The main primary renal diseases causing CRF were acquired renal diseases. Conservative treatment was the main therapy of CRF, but renal replacement therapy was initiated in some of the cases. The obvious difference between follow-up cases and lost cases warrants the need to establish a management system of childhood CRF. PMID- 16221337 TI - [Idiopathic collapsing glomerulopathy in children: report of two cases]. AB - OBJECTIVE: Idiopathic collapsing glomerulopathy (ICG) is a clinically and pathologically distinct variant of focal segmental glomerulosclerosis, which is characterized by proteinuria (often nephrotic range) and rapid progression to end stage renal failure. The typical pathological changes are global/segmental glomerular collapse, hypertrophy and hyperplasia of podocytes and severe tubulointerstitial lesions. Most ICG patients who have been reported in previous published papers are adults. ICG in children is rare. The study aimed to analyze and investigate clinical manifestations, renal histopathological findings, treatment and outcomes of ICG in children. METHODS: Data of two cases of ICG, a 7 year-old boy and a 12-year-old girl, were analyzed. Both of them were Chinese and Han. Clinical characteristics, results of laboratory tests, renal histopathological findings, treatment, outcomes and prognosis of the two children with ICG were retrospectively analyzed. Results were compared with published data. RESULTS: These two children presented typical clinical features of nephrotic syndrome. The quantity of 24 hr urine protein was 7.6 g/d (0.47 g/kg x d for boy) and 10.67 g/d (0.35 g/kg x d for girl). Both of them had hypertension (blood pressure ranged from 130/90 to 150/110 mmHg) and hypercholesterolemia (15.4 mmol/L for the boy and 11.3 mmol/L for the girl). The serum albumin was 12 g/L for girl and 23 g/L for boy. The creatinine clearance rate gradually decreased from normal range to 30 ml/min for the girl. The histopathological changes in renal biopsy of them were focal segmental or global glomerular collapse, hypertrophy and hyperplasia of podocytes and severe tubulointerstitial lesions. These two cases were steroid-resistant and were treated with pulse intravenous methylprednisolone and pulse intravenous cyclphosphamade in one case, who rapidly progressed to end-stage renal failure and died half a year later. Another one was treated with cyclosporine. He showed continuous hypertention and heavy proteinuria for eight months. CONCLUSION: ICG in the 2 children was a severe disease which presented steroid-resistant nephrotic syndrome and rapidly progressive renal failure. The pathological characteristics was global/segmental glomerular collapse, hypertrophy and hyperplasia of podocytes and severe tubulointerstitial lesions. In children with ICG treatment was difficult and the prognosis was poor. PMID- 16221338 TI - [Effect of Lp(a) on human mesangial cell proliferation, adhesion and migration]. AB - OBJECTIVE: The renal disease is commonly associated with hyperlipidemia and correlates with glomerular accumulation of atherogenic lipoproteins and mesangial hypercellularity. Therefore, in this study, the authors investigated a possible growth stimulatory effect and mode of action of lipoprotein(a) [Lp(a)] in human mesangial cells HMC, and the effect of Lp(a) on adhesion and migration in human mesangial cells. METHODS: The DNA synthesis of HMC was measured by (3)H-thymidine incorporation. The cell adhesion was detected by the expression of vinculin by means of indirect immunofluorescence. The cell migration was observed under the microscope. RESULTS: The incubation of HMC with Lp(a) for 24 hours induced a significant dose-dependent proliferation of HMC [Lp(a): 5 microg, 10 microg, 25 microg, 50 microg/ml vs. control 0 microg/ml; (3)H-TdR incorporation (x 10(3)cpm): 1.69 +/- 0.48, 3.59 +/- 0.68, 4.14 +/- 0.78, 4.05 +/- 0.55 vs. 1.64 +/ 0.31, P < 0.01]. The vinculin staining by indirect immunofluorescence showed positive result when HMC was incubated with 10 microg/ml Lp(a) for 24 hours, while vinculin was negative when HMC was incubated with 0 microg/ml Lp(a) as the control of the study. The incubation of HMC with 10 microg/ml Lp(a) for 72 hours demonstrated significant cell migration effect compared to the control of 0 microg/ml. (16.2/LP vs. 2.4/LP, P < 0.01). CONCLUSION: Lp(a) could stimulate a proliferation, adhesion and migration effect on human mesangial cells. PMID- 16221339 TI - [Renoprotective effects of matrine on experimental glomerulosclerosis in rats]. AB - OBJECTIVE: Matrine has an anti-fibrosis effect, such as hepatic cirrhosis and derma fibrosis, while its effect on glomerulosclerosis is unknown. The purpose of this study was to analyze the renoprotective effects of matrine on experimental glomerulosclerosis in rats and inquire into its mechanisms. METHODS: The rats were randomly assigned to following groups: normal control group, model control group, benazepril treatment group, matrine 100 mg/kg treatment group and matrine 50 mg/kg treatment group. The rats of normal control group were subjected to sham operation and were injected with normal saline via the tail vein one week later. The rats of the other groups were uninephrectomized and injected with adriamycin (5 mg/kg) via the tail vein one week later. The dose of benazepril was 6 mg/kg. Both matrine and benazepril were given by gastric perfusion from the first day after the operation. The level of urinary protein was measured at the 2nd, 4th and 6th week after the operation. The serum total protein and albumin, serum creatinine, blood urea nitrogen (BUN) were tested only at the 6th week after operation. Renal pathology changes were evaluated at the 6th week as well. Immunohistochemistry was used to detect the expression of fibronectin (FN), laminin (LN), connective tissue growth factor (CTGF) and transforming growth factor-beta1 (TGF-beta1) in glomeruli. RESULTS: Matrine and benazepril not only reduced the excretion of urinary protein and the level of serum creatinine and BUN, but also significantly ameliorated glomerular mesangial proliferation and glomerular sclerosis (P < 0.05, respectively). Immunohistochemical staining indicated that there was an increasing FN, LN, CTGF and TGF-beta1 expression in model control group as compared to the three treatment groups (P < 0.05). Matrine 100 mg/kg treatment group and benazepril treatment group showed much more advantages than matrine 50 mg/kg treatment group (P < 0.05), but there was no significant difference between the former two groups (P > 0.05). CONCLUSION: Matrine has a renoprotective effect on experimental glomerulosclerosis in rats, the possible mechanism might relate to the reduction of the TGF-beta1 negative function via CTGF, which will inhibit the activation and proliferation of glomerular intrinsic cells, decrease the secretion of ECM accordingly. PMID- 16221340 TI - [Diagnosis of microscopic hematuria in children]. PMID- 16221341 TI - [Pathophysiological investigation on nutrition, infection and bone metabolism in children with chronic renal failure]. PMID- 16221342 TI - [Non-dialysis therapy for chronic renal insufficiency]. PMID- 16221343 TI - [Management of primary vesicoureteral reflux in children]. PMID- 16221344 TI - [Podocyte and proteinuria]. PMID- 16221345 TI - [Progresses in studies on the glucose transporter deficiency syndrome]. PMID- 16221347 TI - [Clinical study of 83 cases with spinal muscular atrophy in children]. AB - OBJECTIVE: Spinal muscular atrophy (SMA) is a common autosomal recessive disorder and represents one of the most common genetic causes of death in childhood. The last 10 years have seen major advances in the field of SMA, but no curative treatment is available so far. This study aimed to analyze the clinical characteristics of SMA, improve the clinical diagnosis of SMA, and explore the importance of gene diagnosis and prenatal diagnosis of SMA by gene deletion analysis. METHODS: Totally 83 cases with SMA including 55 males and 28 females were enrolled in this study. The age was between 1 day and 14 years (average 23.7 months). The clinical characteristics and changes of electromyography were assessed in all cases. The muscular biopsy was performed in 2 of 83 cases. The deletion of survival of motor neuron gene (SMN) was detected by PCR and restriction endonuclease spectrum analysis in 13 of 83 cases. RESULTS: The 83 cases were subdivided into three clinical groups based on age of onset of symptom, age at death and achievement of certain motor milestone, 60 cases with type I, 19 cases with type II and 4 cases with type III. They were all characterized by symmetric muscle weakness (more proximal than distal) associated with atrophy, absence or marked decrease of deep tendon reflexes. Electromyographic studies showed a pattern of denervation with neither sensory involvement nor marked decrease of motor nerve conduction velocities in all cases. Muscle biopsy provided evidence of skeletal muscle denervation with groups of atrophy in 2 cases. The SMN detection revealed deletion of exon 7 and exon 8 in 11 of 13 cases, only lacking exon 7 in 1 of 13 cases and lacking exon 8 in 1 of 13 cases. CONCLUSION: SMA is characterized by degeneration of lower motor neuron associated with muscle paralysis and atrophy. The definite diagnosis of SMA will rely on the typical clinical characteristics, changes of electromyogram and muscle biopsy and gene deletion analysis. Gene diagnosis of SMA can provide a basis for prenatal diagnosis which is of great importance in preventing SMA. PMID- 16221346 TI - [A Chinese girl with cleidocranial dysplasia (CCD) caused by the recurrent R190W mutation in RUNX 2]. AB - OBJECTIVE: Cleidocranial dysplasia (CCD) is a rare skeletal disease with autosomal dominant inheritance associated with mutation in RUNX 2. The authors report a Chinese girl with CCD in whom the mutation in RUNX 2 was identified. METHODS: Clinical diagnosis was based on physical examination, radiological findings, and biochemical tests. For mutation detection, genomic DNA was extracted from peripheral blood using standard method. All 7 coding exons of RUNX 2 and their flanking intronic sequences were amplified by polymerase chain reaction (PCR), and the PCR products were then subjected to automatic DNA sequencing. RESULTS: The affected girl showed typical clinical manifestations of CCD, including patent fontanelles, absent clavicles, short stature and dental anomalies. Direct sequencing of PCR-amplified fragments revealed a recurrent missense mutation, R190W (568 C > T), in RUNX 2. The mutation was further confirmed by Hae III restriction analysis. CONCLUSION: A Chinese case of CCD was confirmed and the disease-causing mutation was linked to a recurrent point mutation in RUNX 2. PMID- 16221348 TI - [Oral motor dysfunction, feeding problems and nutritional status in children with cerebral palsy]. AB - OBJECTIVE: This study was undertaken to investigate the clinical features of oral motor dysfunction and feeding problems as well as the nutritional status of children with cerebral palsy (CP). METHODS: Fifty-nine CP children, 39 boys and 20 girls, mean age 31 months (20 to 72 months), were recruited. Their parents were interviewed for high risk factors and feeding history. Each case was assessed for oral motor and feeding problems based on oral motor and feeding skill score; for nutritional status by measurement of weight, height; neurologically for type of cerebral palsy and for developmental age by Gesell's developmental scale. Equal number of age and sex matched controls were included for comparison of nutritional status, oral motor and feeding skill score. RESULTS: Among 59 patients, 51 cases had oral motor dysfunction and 55 cases had feeding problems including all athtosis, spastic tetraplegia, and 16 had spastic diplegia. The scores of both the mean oral motor function and feeding skill of CP children were significantly lower than those of the controls (P < 0.001). Main food of children with cerebral palsy consisted of liquid and semisolid diet. Body weight and height below the 25th percentile were found in 13 cases and 19 cases, respectively. CONCLUSIONS: The majority of the children with cerebral palsy had oral motor dysfunction and feeding problems which appeared in early age and disturbed the growth and nutritional status. Thorough assessment for oral motor function, feeding problems and nutritional status of CP children is indicated in order to start timely rehabilitation and nutritional interventions which can significantly improve their nutritional status and quality of life. PMID- 16221349 TI - [Prevalence of Helicobacter pylori resistant to clarithromycin, amoxicillin and metronidazole in children]. AB - OBJECTIVE: To investigate the prevalence of Helicobacter pylori (Hp) resistant to clarithromycin, amoxicillin and metronidazole in children. METHODS: A total of 44 Hp clinical isolates were cultured from children patients with Hp gastritis and peptic ulcer who underwent gastroscopy during the period from October 2002 to November 2003 in Children's Hospital affiliated to Medical College of Zhejiang University. Nineteen of the patients were girls and 25 boys. The mean age was 8.66 +/- 2.66 years (3 approximately 14 years). The susceptibilities of the 44 Hp strains to clarithromycin, amoxicillin and metronidazole were tested by agar dilution test to determine minimum inhibitory concentration (MIC)(50), MIC(90) and resistance rates were also calculated. RESULTS: Among the 44 Hp strains isolated from children, 8, 4, and 14 strains were resistant to clarithromycin, amoxicillin and metronidazole and the resistant rate was 18.2% (8/44), 9.1% (4/44) and 31.8% (14/44), with MICs for all strains ranging from < 0.125 microg/ml to 64 microg/ml, < 0.125 microg/ml to 64 microg/ml and < 0.125 microg/ml to 128 microg/ml, respectively. MIC(50) and MIC(90) of clarithromycin, amoxicillin and metronidazole were < 0.125 microg/ml, 8 microg/ml; < 0.125 microg/ml, 0.5 microg/ml; 0.5 microg/ml, 128 microg/ml, respectively. The prevalence of Hp resistant to clarithromycin was higher in children than that in adults, while to metronidazole was lower than that in adults with high MICs (32 - 128 microg/ml) and MIC(90) (128 microg/ml) of metronidazole for resistant isolates. Three multidrug resistant strains (6.8%) were simultaneously resistant to these three antibiotics. CONCLUSIONS: The prevalence of Helicobacter pylori resistant to metronidazole was high in children, and the rate of resistance to clarithromycin was higher than that isolated from adults. Amoxicillin-resistant strains were now found, and multidrug resistant strains which resistant to the three antibiotics also emerged. PMID- 16221351 TI - [Can premature infants be treated with hyperbaric oxygen?]. PMID- 16221350 TI - [Effects of nonnutritive sucking on gastric emptying and gastroesophageal reflux in premature infants]. AB - OBJECTIVE: Although nonnutritive sucking (NNS) during tube feeding has some benefits on the physiology and development of premature infants, the effect on gastrointestinal function remains controversial. The aim of the study was to evaluate the effects of NNS on the gastric emptying and gastroesophageal reflux (GER) in premature infants. METHODS: Thirty eight healthy appropriate-for gestational-age premature infants (birth weight ranged from 1050 g to 1790 g, gestational age ranged from 28 weeks to 35 weeks) accepting intermittent nasogastric feeding (INGF) were randomized into NNS group and N-NNS group according to INGF with and without NNS and fed with the same milk formula. Group NNS (n = 18) received oral stimulation by means of a pacifier immediately before feeding, during feeding and then after feeding for 5 min. Group N-NNS (n = 20) served as control and received INGF alone. The following data were collected and recorded, the fluid intake (including both intravenous and oral), milk intake, caloric intake, time of caloric intake reaching 418.4 kJ/(kg x d) by enteral feeding and relevant condition to feeding. Gastric emptying was measured when oral intake reaching above 8 ml/kg while concurrently measuring 24 hour esophageal pH. Real time ultrasonic images of the gastric antrum were obtained and the antral cross sectional area (ACSA) was measured and the half emptying time (50% DeltaACSA) was calculated. Using 24-hour intraesophageal pH monitoring for evaluation of GER, the five parameters of esophageal pH were recorded: number of reflux episodes during 24 hours, reflux index, number of episodes lasting > 5 min, the duration of longest episode and the total time of pH < 4.0. RESULTS: Within two weeks after feeding, there was no significant difference in the fluid intake, caloric intake between the two groups (P > 0.05). Gastric emptying was measured on day 13.26, milk intake had no difference between the two groups and there was no difference in prefeed ACSA. The half gastric emptying time in NNS group was significantly shorter than that in N-NNS group [(58.33 +/- 22.94) min vs. (73.75 +/- 17.76) min, P < 0.05]. Thirty-two of the 38 infants developed GER, the morbidity was 84.2%; the number of reflux episodes during 24 hours was significantly fewer in NNS group than that in N-NNS group [9 (2 - 31) vs. 14 (5 - 31), P < 0.05]; the total time pH < 4.0 and reflux index was lower in NNS than that in N-NNS, but the difference was not statistically significant. The time of reaching 418.4 kJ/(kg x d) by enteral feeding in NNS group was significantly shorter than that in N-NNS group [(12.36 +/- 4.29) d vs. (15.50 +/- 4.58) d, P < 0.05]. The incidence of feeding intolerance such as vomiting and abdominal distension was lower in NNS group than that in N-NNS group, but the difference was not statistically significant (P > 0.05). However, the morbidity of gastric residue in NNS was significantly lower than that in N-NNS (16.7% vs 50.0%, respectively, P < 0.05). CONCLUSION: NNS used during intermittent nasogastric tube feeding is an easy and safe intervention. NNS can improve gastric emptying and decrease the number of reflux episodes, has a positive improving effect on the development of gastrointestinal motility, is beneficial to premature infants for establishing postnatal enteral nutrition. PMID- 16221352 TI - [Protective effects and mechanisms of recombinant human superoxide dismutase in acute lung injury of rats following meconium aspiration]. AB - OBJECTIVE: To evaluate the protective effects of recombinant human superoxide dismutase (rhSOD) in acute lung injury (ALI) following meconium aspiration. METHODS: Thirty-two healthy male Sprage-Dawley rats were divided into two groups, 8 were used as control (saline group) by infusing 1 ml/kg saline through endotracheal tube; the other 24 rats were used to establish model of ALI by infusing 1 ml/kg of 20% human newborn meconium suspension through endotracheal tube, and then were randomized to 3 groups (8 each): meconium group with no administration of saline or rhSOD; meconium + saline group by infusing 1 ml/kg saline through endotracheal tube; meconium + rhSOD group by infusing 20 mg/kg rhSOD dissolved in 1 ml/kg saline through endotracheal tube. The rats were killed 24 h after treatment. The measurements included bronchoalveolar lavage fluid (BALF) cell counts, protein, BALF protein/plasma protein (pulmonary permibility index, PPI), lactic dehydrogenase (LDH), pulmonary myeloperoxidase (MPO) and superoxide dismutase (SOD) activity, malonyldialdehyde (MDA) and nitric oxide (NO) level. Lung injury score was also evaluated. RESULTS: Compared with the saline group, the rats in the meconium group had significantly increased BALF cell counts (4.04 +/- 1.01 vs. 0.53 +/- 0.19), protein (2.54 +/- 0.74 vs. 0.67 +/ 0.26), PPI (0.50 +/- 0.18 vs. 0.12 +/- 0.05), LDH (263.50 +/- 97.84 vs. 17.38 +/ 3.58), pulmonary MPO (1.49 +/- 0.22 vs. 0.62 +/- 0.16), MDA (3.30 +/- 0.85 vs. 1.40 +/- 0.35), NO (12.77 +/- 5.00 vs. 4.89 +/- 1.32) and lung injury score (9.88 +/- 2.10 vs. 2.25 +/- 1.04), P < 0.01 for all, whereas pulmonary SOD activity had no statistically significant differences (103.28 +/- 24.53 vs. 94.49 +/- 12.93, P > 0.05). There were no statistically significant differences between meconium + saline group and meconium group (all P > 0.05). Compared with the meconium + saline group, meconium + rhSOD group had decreased BALF cell counts (3.13 +/- 0.77 vs. 4.68 +/- 1.40, P < 0.01), LDH (162.63 +/- 76.90 vs. 273.75 +/- 111.83, P < 0.05), pulmonary MPO activity (1.23 +/- 0.28 vs. 1.54 +/- 0.24, P < 0.05), MDA (2.46 +/- 0.42 vs. 3.50 +/- 0.82, P < 0.01), NO level (9.17 +/- 2.34 vs. 13.04 +/ 4.38, P < 0.05), lung injury score (8.63 +/- 1.30 vs. 10.00 +/- 1.07, P < 0.05) and increased pulmonary SOD activity (134.45 +/- 23.30 vs. 106.79 +/- 17.77, P < 0.05), but there were no statistically significant differences in BALF protein and PPI between these two groups. CONCLUSION: Inflammation and lipid peroxidation might play important roles in the pathogenesis of ALI with meconium aspiration, a single early administration of 20 mg/kg rhSOD intratracheally can reduce lung damage in rats following meconium aspiration. PMID- 16221353 TI - [Effect of different early nutritional interventions on catch-up growth of rats with intrauterine growth retardation]. AB - OBJECTIVE: About 20 - 50% individuals with intrauterine growth retardation (IUGR) could not achieve catch-up growth and remain small in size till adulthood. There are few reports on the relation between intestinal development and body catch-up growth of IUGR. Studies showed that early "nutritional programming" would results in long-term effects on the body growth and organic function, and gastrointestinal development is closely related to the body development as well. The authors aimed to study the effect of early nutritional interventions on serum IGF1, IGFBP3, intestinal development and catch-up growth of pups with IUGR by using diets with different protein and caloric levels during the first four weeks of life. METHODS: An IUGR rat model was established by maternal nutrition restriction during pregnancy. Thirty-two IUGR female pups were divided randomly into 4 groups (8 pups in each group) and eight normal female pups as control. The groups and interventions were (1) Normal control group (C group); (2) IUGR control group (S group), (3) IUGR low-protein diet group (SL group); (4) IUGR high-protein diet group (SH group); (5) IUGR high-caloric group (SA group). The serum IGF1, IGFBP3, body weight, body length, and intestinal weight, length, intestinal villi height (VH), crypt depth (CD), villi absorbing area (VSA), mucous thickness (MT) were measured at the 4(th) week of life. RESULTS: (1) At the 4(th) week, the serum IGF1 (724.0 +/- 153.5 ng/ml), IGFBP3 (9.69 +/- 3.13 ng/ml), and VH (416.9 +/- 46.3 microm), VSA (115.9 +/- 24.0 x 10(3) microm(2)), MT (583.9 +/- 68.5 microm) in the SH group were significantly higher than those of normal control group (539.4 +/- 198.4 ng/ml, 4.77 +/- 2.98 ng/ml and 322.1 +/- 25.8 microm, 85.8 +/- 17.8 x 10(3) microm(2), 480.0 +/- 61.5 microm) and IUGR control group (P < 0.05). The intestinal weight (1.91 +/- 0.16 g) and length (80.67 +/- 9.47 cm) in the SH group was not significantly different from the normal control group (2.24 +/- 0.22 g and 74.77 +/- 9.06 cm, P > 0.05). The SH group showed the fastest catch-up growth. Their body weights (40.14 +/- 11.03 g) at the 3(rd) week and body lengths (23.61 +/- 0.49 cm) at the 4(th) week of life reached the normal ranges of the control group (44.65 +/- 5.36 g and 23.10 +/- 1.42 cm, P > 0.05). (2) The serum IGF1 (346.7 +/- 85.3 ng/ml), IGFBP3 (1.4 +/- 0.21 ng/ml), body weight (21.41 +/- 3.54 g) and body length (15.96 +/- 1.29 cm) and the most of intestinal indexes in the SL group were markedly lower than other groups at the 4(th) week of life (P < 0.05). CONCLUSION: The serum IGF1 was a sensitive marker to reflect the catch-up growth and nutritional status, and IGF1 was positively correlated with the intestinal development and body growth. When given different nutritional interventions during the first four weeks of life, high protein diet is more helpful for the IUGR catch-up growth by promoting the intestinal development and the absorption of nutrition. PMID- 16221354 TI - [The twenty one clinical of children's mdinitrobenzene collective poisoning]. PMID- 16221355 TI - [Effects and significance of blockade of renin-angiotensin system on the progression of renal injury in rats with glomerulosclerosis]. PMID- 16221356 TI - [Vitamin D receptor gene polymorphism in patients with rickets]. PMID- 16221357 TI - [Plasma exchange of two children with thrombotic microangiopathy]. PMID- 16221358 TI - [Analysis of misdiagnosis of hemorrhagic fever with renal syndrome in 4 young children]. PMID- 16221359 TI - [White matter pattern of Leigh's syndrome, a case report]. PMID- 16221360 TI - [Antiepileptic medication induced increase of seizure attacks: report of 6 cases]. PMID- 16221361 TI - [Transcatheter closure of aneurysm of perimembrane ventricular septal defect with Amplatzer duct occluder in children]. PMID- 16221362 TI - [A case of infantile neuroaxonal dystrophy]. PMID- 16221363 TI - [Report of a case with neonatal cardiac hemangioma]. PMID- 16221364 TI - [A case of prune belly syndrome]. PMID- 16221365 TI - [Cartilage-ectodermal dysplasia in a case]. PMID- 16221367 TI - Pediatric traumatic brain injury: beyond the guidelines. AB - In 2003, a multidisciplinary group of physicians formulated the first guidelines for the management of severe traumatic brain injury in infants and children. Initial treatment of these patients is focused on stabilization to prevent the occurrence of secondary insults such as hypotension and hypoxemia. However, this article focuses on the established and emerging therapies used in the intensive care unit management of intracranial hypertension--which represents the key target for contemporary therapy of this condition. A critical pathway for the treatment of this condition was established within the guidelines, and this pathway is appropriately focused on limiting intracranial hypertension and optimizing cerebral perfusion during the intensive care unit phase. This includes first- and second-tier therapies. This article contains a brief synopsis of this critical pathway and discusses important new developments for the management of this condition. Key new developments include a better understanding of the optimal cerebral perfusion pressure target for intracranial pressure-directed therapy, with emerging evidence supporting the use of two therapeutic modalities, mild-moderate hypothermia and decompressive craniectomy. PMID- 16221368 TI - Induced hypothermia: a novel neuroprotective treatment of neonatal encephalopathy after intrapartum hypoxia-ischemia. AB - The current treatment approach to an infant at risk for progression to neonatal encephalopathy after intrapartum hypoxia-ischemia is threefold: 1) early identification of the infant at highest risk for evolving brain injury based on the criteria of a sentinel event during labor, prolonged depression at birth with the need for resuscitation, and evidence of severe fetal acidemia based on a cord umbilical arterial pH less than 7 and/or a base deficit more than -16 meq per L, with early clinical and/or electroencephalogram assessment of moderate to severe encephalopathy; 2) supportive therapy instituted to maintain adequate ventilation and in particular pCO2 levels in a normal range, mean arterial blood pressure within a normal range so as to avoid perturbations in cerebral perfusion, glucose in a normal range to avoid hypoglycemia, and the judicious treatment of seizures; and 3) neuroprotection--induced hypothermia is currently the only strategy that has been rigorously evaluated in two large, multicenter randomized studies. The first study used selective cooling with a cool cap to a rectal temperature at 34.5 degrees C, and the second study used total body cooling to an esophageal temperature of 33.5 degrees C, with the temperature in both studies maintained for 72 hours. No significant side effects were noted with this degree of cooling. The combined data indicate that hypothermia is associated with a reduction in the incidence of death and/or severe disability at 18 months follow-up, with the most significant effect observed in infants who at the initiation of therapy present with modest encephalopathy and/or do not exhibit electrographic seizures. PMID- 16221369 TI - Treatment of pediatric and adolescent headaches. AB - The treatment of childhood headaches requires a thorough investigation of the underlying etiology, including the use of standardized diagnostic criteria, and neurologic and comprehensive examinations. If secondary headaches are identified, the headaches should resolve with treatment of the underlying cause. If the headaches persist or primary headaches are identified, a three-component treatment approach may need to be developed. This includes acute therapy, preventative therapy, and biobehavioral therapy. All of these components need to be addressed in the treatment of childhood headaches, and clear goals of treatment must be discussed with the patient and parents. PMID- 16221366 TI - Hypertonia in children: how and when to treat. AB - Hypertonia in children can be caused by many different diseases. The most common etiology is cerebral palsy. Spasticity and dystonia are the most common types of hypertonia. There are few options for treatment, and usually treatment has an incomplete effect. Therefore, it is necessary to prioritize goals in order to improve overall functional outcome. The use of any intervention will require the ability to verify the magnitude and importance of the outcome in order to ensure that therapy is efficacious. In general, a complex, flexible, and multifaceted approach will be necessary to improve the motor abilities of children with hypertonia. PMID- 16221370 TI - Treatment and prevention of cerebrovascular disorders in children. AB - Cerebrovascular disorders are an important cause of mortality and chronic morbidity in children. Ischemic stroke is more common than cerebral venous thrombosis and hemorrhagic stroke in children. Several medical disorders have been associated with stroke in children, and a thorough evaluation of underlying causes is needed to determine the best treatment and prevention strategy. The treatment and prevention of stroke in children is not well studied, and current recommendations are based on adult studies, nonrandomized trials, or expert opinion. Children with stroke require immediate, special attention and if possible should be stabilized and transferred to an institution that can offer pediatric neurovascular expertise and care. All children with stroke should be referred to or have their care managed by a pediatric neurologist. The treatment of stroke in adults is well studied, and when applicable this evidence should be considered in the treatment of children with stroke. Data from animal and adult stroke studies have demonstrated a benefit for the aggressive treatment of infection, fever, blood pressure, hypo/hyperglycemia, intracranial pressure, and seizures, and should be applied to children with stroke. The use of thrombolytic, antithrombotic, and antiplatelet therapies is based on adult studies, cohort studies, and/or expert opinion. Two consensus guidelines regarding the treatment of arterial ischemic stroke and cerebral venous thrombosis were recently published and recommend the use of anticoagulants or antiplatelet agents in the acute setting, depending on the underlying cause of stroke. The evidence for the primary prevention of stroke in children is restricted to sickle cell disease (SCD) and derived from the Stroke Prevention in Sickle Cell Study Project studies. Long-term chronic transfusion therapy to maintain hemoglobin S levels below 30% is indicated in children with SCD and intracranial stenosis. It has also been recently determined that chronic transfusion therapy should not be stopped in children with SCD and an increased risk for stroke. The recurrence rate of arterial ischemic stroke (AIS) in children ranges from 6% to 30% and is highest among children with recurrent transient ischemic attack, cardiac disease, arteriopathies, and metabolic and coagulation abnormalities. Recommendations for secondary prevention are based on adult studies and the underlying pathophysiology of the stroke. Antiplatelet therapy (aspirin 1-5 mg/kg/day) is recommended in most children with a history of AIS. Although there is minimal evidence to support its use in children, anticoagulation may be indicated in AIS associated with extracranial arterial dissection, prothrombotic disorders, cardiac disease, severe intracranial stenosis, and recurrent AIS while on antiplatelet therapy. PMID- 16221371 TI - The management of patients with patent foramen ovale and stroke. AB - The management of patients with patent foramen ovale (PFO) and stroke remains uncertain. What is known is that PFO is a very common finding in the general population. The risk of initial and recurrent stroke in individuals with PFO is relatively low. With the advent of transesophageal echocardiography, PFO is more commonly being diagnosed. There are insufficient data to support a benefit of warfarin over aspirin in preventing recurrent stroke in most patients with PFO. In those with PFO and a history of a procoagulant state or deep venous thrombosis, anticoagulation may be indicated after weighing the risks of anticoagulation against potential benefits in preventing recurrent stroke. PFO closure does not guarantee the prevention of future stroke and should be addressed case by case, with consideration of patients for entry into ongoing clinical trials of safety, efficacy, and durability. PMID- 16221372 TI - Atrial fibrillation and stroke: four treatment controversies. AB - Atrial fibrillation is an important cause of disabling ischemic stroke, and adjusted-dose warfarin is highly effective for prevention and remains the therapy of choice for high-risk patients. Ximelagatran, a novel oral anticoagulant, is clinically equivalent to warfarin for preventing stroke in patients with atrial fibrillation, but concerns about potential hepatic toxicity have precluded US Food and Drug Administration approval. Many patients with low-risk atrial fibrillation do not benefit substantially from anticoagulation, and these patients can be reliably identified using the CHADS2 stroke risk stratification scheme. A target International Normalized Ratio (INR) range of 2 to 3 is usually recommended for anticoagulation of patients with atrial fibrillation, but a lower INR target (2, range 1.6-2.5) may be a reasonable benefit/risk trade-off for primary prevention in those aged older than 75 years. Adding aspirin to adjusted dose anticoagulation increases bleeding (including central nervous system bleeding), is of uncertain value, and should only be done after careful consideration and with vigorous efforts to control blood pressure. PMID- 16221373 TI - New treatments in acute ischemic stroke. AB - Since the advent of intravenous thrombolytic therapy with recombinant tissue plasminogen activator (tPA) for acute ischemic stroke, there has been a marked change in our management approach to patients with acute ischemic stroke. Although the major part of our focus in treating patients with stroke remains prevention of complications post-stroke and reduction of stroke recurrence, there is a paradigm shift to immediate "clot" lysis. This concept is being actively promoted through certification of institutions as stroke centers in order to increase the number of patients with stroke treated in an ultra-rapid fashion. However, options for acute treatment remain limited. Other than aspirin, the only US Food and Drug Administration-approved agent for acute ischemic stroke is intravenous tPA. Some physicians treating patients with acute ischemic stroke still frequently use heparin and low-molecular-weight heparinoids, but there are no firm data to support routine use of this drug class. However, a number of new lytic agents and strategies are being pursued. Some of these treatments, such as intra-arterial chemical thrombolysis or mechanical intra-arterial thrombolysis, are available only at specialized stroke centers. In addition, new antithrombotic agents are being studied. Drugs that can rescue neurons from impending hypoxia ischemia cell death represent the "holy grail" of acute stroke therapy. To date, these "neuroprotectant" strategies have been unsuccessful, although this concept remains under active investigation in animal and human trials. PMID- 16221374 TI - A "snapshot" of an ovarian cancer clinical practice: evidence for viewing the malignancy as a "chronic disease". PMID- 16221375 TI - New agents in the treatment of childhood leukemias and myelodysplastic syndromes. AB - Relapsed or refractory leukemia remains the most common therapeutic problem in pediatric oncology. Particularly challenging is the patient who has recurrence following a stem cell transplant. Insights into the molecular pathogenesis of the leukemias have produced an array of new agents. These new agents will be more selective in hitting their targets, and so their use will be more narrowly defined than with classical cytotoxic drugs. These new agents include all-trans retinoic acid, gemtuzumab ozogamicin, imatinib mesylate, rituximab, and a bevy of signal transduction inhibitors and therapeutic monoclonal antibodies. Other new agents, such as liposomal daunorubicin, PEG-asparaginase, or clofarabine, represent chemical modifications of established antileukemic drugs. Increasingly, molecular profiling will be used to guide the development and application of new drugs. PMID- 16221376 TI - Umbilical cord blood transplantation. AB - Over the past decade umbilical cord blood has been established as a viable source of hematopoietic stem cell for allogeneic transplantation. Early experience with umbilical cord blood transplantation (CBT) demonstrated a lower incidence of graft-versus-host disease even though the procedure was performed with HLA disparate grafts. The overall outcome of CBT appears similar to that of allogeneic bone marrow transplant. The expansion of the donor selection is particularly beneficial to ethnic minorities, whose representation in the marrow registries is relatively small. The major drawbacks of CBT are slow hematopoietic recovery and a high incidence of graft failure, as a result of a lower number of progenitors infused. This paper reviews the current results of CBT and ongoing investigations to increase its availability to a larger number of recipients. PMID- 16221377 TI - New paradigms for therapy for osteosarcoma. AB - Osteosarcoma is a primary bone malignancy generally affecting the young, with 60% of cases occurring before the age of 25 years and the peak incidence at 15 years. Survival has improved over the past several decades, with non- metastatic disease having an approximately 70% chance of long-term survival. Unfortunately, patients with metastatic disease at diagnosis or those who have recurrent disease have a dismal prognosis, with approximately 20% surviving long term. In this review article we describe several new therapies in development for osteosarcoma. These include immune-based therapies, strategies to inhibit tumor growth, radiotherapy, and the introduction of new chemotherapies and targets. PMID- 16221379 TI - Gynecologic oncology group trials of chemotherapy for metastatic and recurrent cervical cancer. AB - Because only 16% of patients with metastatic cervical cancer are alive 5 years after diagnosis, the Gynecologic Oncology Group (GOG) has carefully designed and conducted many phase II studies to identify promising drugs. Cisplatin has emerged as the most active single agent with overall response rates of 19%. Recent phase III trials have documented response rates of 27% and 39% when cisplatin has been combined with either paclitaxel or topotecan, respectively. The comparison of cisplatin to cisplatin plus topotecan in GOG-179 has yielded the first study to show a statistically significant impact on the overall response rate, median progression-free survival, and median survival, with all outcome measures favoring the two-drug regimen. Despite these encouraging results, however, most of the responses are partial and of short duration. The need for novel combinations and the implementation of active biologic agents is implicit. The accumulated data in this disease setting, as evidenced by the experience of the GOG, are presented in this review. PMID- 16221380 TI - Lymphatic mapping and sentinel node detection in gynecologic malignancies of the lower genital tract. AB - The utility of lymphatic mapping and sentinel lymph node biopsy in malignancies of the female lower genital tract-- vulvar, vaginal, and cervical cancers--is being explored in multiple centers internationally. For patients with these tumors, lymphatic mapping with sentinel lymph node biopsy holds the promise of increasing the identification of microscopically metastatic disease while decreasing the morbidity of complete lymphadenectomy. In this review article we present the published data on mapping techniques and discuss the advantages and pitfalls of these procedures. PMID- 16221382 TI - Secondary cytoreduction for patients with recurrent ovarian cancer. AB - Standard treatment for advanced epithelial cancer includes primary cytoreductive surgery followed by combination chemotherapy. Optimal primary debulking is associated with improved clinical response rates to primary chemotherapy and longer overall survival. Still, at least 60% of advanced-stage ovarian cancer patients who are without clinical evidence of disease after completing primary therapy ultimately develop recurrent disease. Despite significant efforts, standard treatment of patients with recurrent ovarian cancer remains poorly defined. In view of the new refinement in surgical techniques and medical treatments and the extended disease-free interval, physicians and patients face the dilemma of how to handle further management in this subgroup of patients. A clear understanding of the prognostic factors and expectations from the procedure are necessary in order to identify candidates adequately. PMID- 16221381 TI - The merits of vascular targeting for gynecologic malignancies. AB - Neovascularization is an early and critical step in tumor development and progression. Tumor vessels are distinct from their normal counterparts morphologically as well as at a molecular level. Recent studies on factors involved in tumor vascular development have identified new therapeutic targets for inhibiting tumor neovascularization and thus tumor progression. However, the process of tumor blood vessel formation is complex, and each tumor exhibits unique features in its vasculature. An understanding of the relative contribution of various pathways in the development of tumor vasculature is critical for developing effective and selective therapeutic approaches. Several such agents are currently in clinical trials, and many others are under development. In this review, the mechanisms and factors involved in tumor blood vessel formation are discussed. In addition, selected novel classes of antivascular therapies, including those targeting tumor endothelial cells and other components of the tumor vasculature, are summarized. PMID- 16221383 TI - Quality of life and gynecologic malignancies. AB - In 2005, an estimated 79,480 new cases of gynecologic malignancies will be diagnosed, and almost 29,000 women will die. Treatment of gynecologic malignancies involves a combination of surgery, chemotherapy, and/or radiotherapy, all of which may affect a woman's physical, social, and emotional well-being. Although specific effects on quality of life (QOL) depend on disease site, treatment-, and patient-specific factors, common QOL issues include changes in physical functioning due to side effects of treatment, psychological distress due to fear and anxiety of recurrence, sexual dysfunction associated with anatomic and physiologic changes of treatment, and, for younger women, loss of childbearing potential. As new diagnostic and treatment strategies for gynecologic malignancies are developed, research efforts should include QOL consequences. Further studies are needed to develop strategies for identifying women at risk for serious QOL disruption so that effective interventions to assist these women can be designed. PMID- 16221384 TI - How to establish a high-risk cancer genetics clinic: limitations and successes. AB - The development of technology to locate and isolate cancer susceptibility genes has brought together the fields of oncology, cancer control, genetics, and genetic counseling to create a new specialty of cancer risk counseling with the goal to communicate more accurate information about personal cancer risk profiles based on personal and family histories. As cancer risk assessment and counseling services become standard of care in medical practice, their availability is increasingly moving from comprehensive cancer centers and academic institutions to community settings. High-risk cancer genetics clinics in the community face several challenges, including staffing, time commitment, costs, and unique quality control issues. The societal benefits include a more educated public armed with the information needed to make health decisions appropriate for the individual level of risk. PMID- 16221385 TI - What are SNPs and haplotypes and how will they help us manage the prevention of adult cancer? AB - Human genetic variation data are now publicly available on a large scale, from public and private discovery efforts. Datasets from the International Haplotype Map Consortium and Perlegen Sciences provide a level of knowledge about human genetic variation that is unprecedented. In combination with novel high throughput genotyping technologies, these new resources will allow cancer prevention investigators to identify in a more precise way which genetic subsets of patients are likely to benefit most from chemoprevention and screening interventions. PMID- 16221388 TI - [Should badly documented techniques be introduced?]. PMID- 16221389 TI - [Surgery on articular cartilage: from debridement to tissue engineering]. AB - Articular cartilage is a complex tissue with a limited endogenous repair capacity. Surgical strategies to stimulate repair of damaged cartilage are presently evolving extensively. New techniques based on tissue-engineering principles with cultured cells and scaffolds are challenging established techniques based on generating a repair response from the bone marrow. This paper reviews treatments such as debridement, marrow-stimulating techniques, osteochondral transplantation and autologous chondrocyte transplantation. Debridement and marrow stimulation can provide significant relief of symptoms. Recently, randomized studies have uncovered some of the clinical effects of the new biotechnology-based surgical methods. However, no significant advances over the established surgical techniques have been developed. PMID- 16221390 TI - [Risk factors for osteoarthritis of the knee: current status]. AB - The development of arthrosis is best understood paradigmatically as the result of an interaction between individual and systemic risk factors. This review of the current literature on the subject identifies overweight, occupational exposure to repeated daily lifting and the postmenopausal decline in gonadal steroids in women as significant risk factors for the development of symptomatic and radiologic osteoarthritis of the knee. PMID- 16221391 TI - [Uric acid as a marker of pathophysiological mechanisms in patients with cardiovascular disease]. AB - Uric acid can serve as a marker of pathophysiological mechanisms in patients with cardiovascular disease, which may explain why serum uric acid contains prognostic information. Uric acid is the end product of purine degradation. It is catalyzed by the enzyme xanthin oxidase, which is responsible for the production of uric acid and damaging free radicals. This is a central link in the association between serum uric acid and myocardial ischemia, myocardial dysfunction and non cardial function, which is determined primarily by impaired peripheral blood flow. Inhibition of xanthin oxidase with allopurinol may be used in future treatment of heart patients. PMID- 16221392 TI - [Congenital heart block and autoantibodies]. AB - Maternal IgG autoantibodies against Ro52 (part of the anti-SSA specificity) are correlated with congenital heart block (CHB). Most cases revert spontaneously and can be monitored by Doppler echocardiography, but fetuses that progress into second-degree block require treatment with, e.g., corticosteroids. A third-degree block is irreversible and necessitates lifelong pacemaking. It may prove beneficial to screen all expectant mothers for anti-Ro52 autoantibodies. Decisions about this will require further studies of the diagnostic performance of the Ro52 autoantibody test. PMID- 16221393 TI - [Should the sensible drinking limits for adults be changed?]. AB - The sensible drinking limits may be too high depending on sex, age, genetic variation and other factors. Recent research on the alcohol-related risk of breast cancer and the impact of drinking patterns on health calls for an update of the literature. We focus on studies of the effect of alcohol on morbidity and mortality and conclude that the maximum of 21 drinks per week for men and 14 for women may be maintained. We emphasize that episodic intake of alcohol increases the risk of death and that the beneficial effect of alcohol is reached at an intake of one drink per day. We also specify groups for whom the drinking limits are not valid. PMID- 16221394 TI - [A simple model for training basic laparoscopic skills]. AB - We have designed a new kind of black box for training laparoscopic skills. The model is simple and cheap, which makes the opportunities for training much more available. The design is based on the use of a webcam. The model is not a closed box but a table with the camera placed underneath, transmitting the image of the task being performed up onto a computer placed where the surgeon can see it. The open model makes lights and optics unnecessary. We recommend this model for basic laparoscopic training. PMID- 16221396 TI - [Renal and urinary calculi in children]. AB - INTRODUCTION: The incidence of nephrourolithiasis amongst children in Denmark has not previously been recorded. This article reports that incidence, investigations and the treatment prescribed. MATERIALS AND METHODS: During the period 1994-2003, 32 cases were admitted, diagnosed and treated in two Danish paediatric departments. All cases were reviewed retrospectively. RESULTS: All patients underwent radiological examination, such as ultrasound or renography. All the children underwent different degrees of metabolic evaluation. Kidney stone analysis was performed in 14 of the 32 cases (44%). In 21 children (66%) there were one or more predisposing factors to stone formation, the most common being urinary tract malformation, found in 40% of the cases, of which 42% presented with hypercalciuria. DISCUSSION: The various causes of nephrolithiasis in children are discussed, especially the need for clarifying the underlying metabolic causes. The article concludes by recommending an examination programme for children with nephrourolithiasis. PMID- 16221395 TI - [Central nervous system tumours in children. An evaluation of the completeness and validity of the Cancer Registry]. AB - INTRODUCTION: The aim of this study was to evaluate the registration of childhood CNS tumours in the Danish Cancer Registry (CR) from 1980 to 1996, based on completeness and validity. MATERIALS AND METHODS: The completeness of the CR was estimated by examination of the Danish National Hospital Register and an independent hospital registry. To determine the validity of the CR, 640 cases of childhood CNS tumours identified in the CR were compared with the corresponding medical record, with regard to cancer morphology and date of diagnosis. RESULTS: The completeness of the CR was 98-99%. For 82% of all cases, the morphology registered in the CR was identical with that in the medical record. The validity of the morphological diagnosis was 84% for astrocytomas, 88% for ependymomas, 95% for medulloblastomas, 47% for other gliomas and 76% for miscellaneous intracranial and intraspinal neoplasms. In a further 7% of all cases, the morphological diagnosis in the CR was incorrect with respect to details but correct within these main diagnostic groups, 6% were tumours of another main diagnostic group than that registered in the CR and 5% were not tumours, were tumours outside the study criteria or were registered in the hospital journal as suspected tumours. The date of diagnosis in the CR and that in the medical records were concordant in 88% of the evaluated cases. CONCLUSION: Registration of childhood CNS tumours in the CR was highly complete, while the validity of the morphological diagnosis differed among the main diagnostic groups. The relatively low validity might be explained by the anatomic location of CNS tumours, which makes tissue sampling for histological verification and therefore diagnosis difficult. PMID- 16221397 TI - [Should health care information be provided in more than words? Experiences of plastic surgery in patients at looking at clinical photos]. AB - INTRODUCTION: This article deals with an empirical study on the use of clinical photos as part of the information/communication between health professionals and patients. Normally the verbal method is used to provide information to patients. The hypothesis is that figurative and illustrative material can be a useful supplement to the spoken word. The study focused on patients' experience of looking at clinical photos. MATERIALS AND METHODS: The research was conducted in the Department of Plastic Surgery and Burn Care, Rigshospitalet, Copenhagen, Denmark. The study was a quality, explorative and descriptive interview research including 10 patients. RESULTS: This study shows that patients find it positive to look at clinical photos, because it gives them knowledge and understanding, as well as realistic expectations and a sense of security in having made the right decision. The photographs help confirm or invalidate their expectations. The combination of listening to verbal information and looking at pictures makes sense to the patient. DISCUSSION: This study shows that looking at clinical photos is one way to improve the information/communication between patient and health professionals. Clinical photos contribute to the emotional and bodily part of gaining knowledge and thereby complement the verbal information, which is primarily cognitive. The study shows that patients experience photos as a positive element in their treatment and demonstrates that photos give them an answer to how they will appear after surgery, which is difficult to describe in words. In this study age, sex and diagnosis did not seem to matter for the experiences of the patients. PMID- 16221398 TI - [Valproate-caused encephalopathy]. AB - Valproate is a widely used antiepileptic drug with rarely occurring serious side effects. We observed two cases of valproate-caused encephalopathy of insidious type. Ten months after initiation of valproate treatment, the two patients showed neurologic deficits such as rapid cognitive decline and extrapyramidal manifestations. However, the patients recovered some weeks after termination of the treatment. This side effect must be considered in patients showing signs of rapidly progressing severe dementia who are being treated with valproate. PMID- 16221399 TI - [Serious postoperative metabolic alkalosis during hospitalization]. AB - Metabolic alkalosis is a frequent and serious abnormal acid-base disturbance with a high-pH-dependent morbidity and mortality rate. This situation is easy to recognize, and the treatment is relatively simple. We present a case history dealing with metabolic alkalosis as a consequence of severe acid-base disturbance with electrolyte and fluid imbalance caused by continuous postoperative vomiting and nasogastric suction. Our aim is to attract greater attention to this acid base disturbance and a better understanding of the pathogenesis, which in turn should lead to earlier intervention and more appropriate therapy. PMID- 16221407 TI - [A reflection over craniosacral therapy]. PMID- 16221408 TI - [Recovery--an antithesis to the medical scientific approach to mental disease?]. PMID- 16221423 TI - [Diagnostic imaging in acute pulmonary embolism]. PMID- 16221424 TI - [Epilepsy as a dynamic condition: facilitation and inhibition, establishment and remission]. AB - Epilepsy should not be seen as a stable chronic illness but as a condition that undergoes changes over time, due to antagonistic factors which facilitate or counteract seizures. Some of these are known, and their action can best be observed at onset, when seizure repetition is established or prevented, and at remission, when the decline of the antiepileptic drug therapeutic threshold reflects a fall in the propensity to seizures. A quantitative model is proposed which can explain much of the observed variability. PMID- 16221425 TI - [Radioimmunotherapy in non-Hodgkin's lymphoma. With focus on anti-CD20 treatment]. AB - Radioimmunotherapy in indolent NHL is a new and promising therapy which adds a new therapeutic tool to the treatment palette. Recent advantages in RIT are described and perspectives discussed. PMID- 16221426 TI - [Diagnostic imaging in acute pulmonary embolism. The use of spiral computed tomography, lung scintigraphy and echocardiography]. AB - Acute pulmonary embolism is an underdiagnosed and potentially lethal condition. Treatment may be lifesaving but is associated with severe side effects. Thus, reliable diagnostic imaging is essential. We conducted a literature review on the use of spiral computed tomography, lung scintigraphy and echocardiography in acute pulmonary embolism and identified 562 articles, of which 16 original papers met our inclusion criteria. From these, we concluded that none of the modalities is applicable in every situation. Spiral computed tomography can confirm the diagnosis but cannot rule out subsegmental embolism. With lung scintigraphy, perfusion imaging alone is probably sufficient and suited to both confirming and ruling out the diagnosis. Echocardiography should be reserved for patients with an emergent need for treatment and cannot rule out the diagnosis. PMID- 16221427 TI - [Diagnostic imaging in acute pulmonary embolism in Denmark. A survey]. AB - INTRODUCTION: Acute pulmonary embolism is a frequently occurring disease that is associated with a high mortality rate. Accurate diagnosis is essential, but the condition is often misdiagnosed. National and international guidelines do not agree on which imaging modality is preferable for making the diagnosis. Our objective was to determine what the diagnostic method of choice in Denmark is. MATERIALS AND METHODS: We conducted a survey in 30 hospital departments in Denmark regarding the primary imaging procedure for patients with suspected pulmonary embolism. These departments represented various types of hospitals in all parts of Denmark but were otherwise randomly selected. RESULTS: Twenty-seven departments responded. Approximately half of the departments used lung scintigraphy as the primary diagnostic test, one third used echocardiography and the rest used spiral computed tomography. This distribution was largely independent of the type of hospital and which modalities were available in a given hospital. DISCUSSION: We found no coherence between the primary diagnostic modality of choice and other factors including hospital size, type and availability of diagnostic methods. PMID- 16221428 TI - [Does evidence lead to a change in clinical practice? Danish anaesthetists' and nurse anesthetists' clinical practice and knowledge of postoperative residual curarization]. AB - BACKGROUND: Recent studies have shown a high incidence of postoperative residual curarization (PORC). The reason for this is unclear. The purpose of this study was to evaluate whether the methods used by Danish anaesthetists to exclude PORC are evidence-based and, if they are not, to determine the reasons why. METHODS: 251 anaesthetists (nurses and physicians) from eight different hospital anaesthetic departments completed a questionnaire concerning their knowledge about and clinical practice of residual curarization. RESULTS: All of the 251 participants filled in the questionnaire. 91% underestimated the incidence of PORC after the administration of intermediate-acting muscle relaxants, and 27% incorrectly believed that it is always possible to exclude PORC using clinical tests. Only 45% knew that the train-of-four ratio must exceed 0.9 to exclude residual curarization, and only 25% knew that clinically significant residual curarization cannot be excluded by tactile/visual evaluation of the response to train-of-four nerve stimulation. 91% had access to a nerve stimulator, 85% with the option of objective monitoring. 13% used a nerve stimulator seldom or never. The overall attitude toward using nerve stimulators was positive. CONCLUSION: The results indicate that Danish anaesthetists" clinical practice regarding residual curarization is often not evidence-based. The reason for this seems to be a lack of knowledge, rather than lack of resources and/or a negative attitude toward neuromuscular monitoring. PMID- 16221429 TI - [Treatment of young patients with type 1 diabetes in an adult diabetes clinic]. AB - INTRODUCTION: It is well known that children and adolescents with type 1 diabetes mellitus (DM) treated in outpatient clinics often have poor metabolic control and thereby a high risk of developing microvascular complications. Our aim was to determine the status of the young type 1 DM patients under 25 years of age attending our adult outpatient diabetes clinic. MATERIALS AND METHODS: Retrospectively, we registered HbA1c, screening for complications, success with self-care and non-attendance. We suggest criteria for determining the success of the treatment process and outcome. RESULTS: Thirty-four patients, age 22.2 +/- 2.7 years, attend the clinic. Their latest HbA1c was 9.2 +/- 1.9% (mean SD); 18% had HbA1c pound 7.5%. Intensive insulin therapy was used by 91%. Five patients had either background retinopathy, elevated urinary albumin to creatinine ratio or hypertension ( 130/80) on one occasion within the last two years. Dietary problems were listed in 29% of the records, and insufficient frequency of blood glucose measurements appeared in 29%, while 38% had between three and seven non attendances. The patients in all three of these groups had significantly higher HbA1c. Screening for complications was successful according to the criteria for blood pressure, HbA1c and retinopathy, but not completely successful for nephropathy and neuropathy. CONCLUSION: The young type 1 diabetes patients attending our adult outpatient clinic have an unsatisfactorily high HbA1c. Screening for complications is sufficient, and hardly any of the patients have signs of complications, but the rate of non-attendance is high, and there are significant problems with self-care. New individual considerations for these young patients appear to be necessary and standards for the quality of treatment are needed. PMID- 16221430 TI - [The clinical examination of the shoulder]. PMID- 16221432 TI - [Don't always rely on PACS: undetected loss of a central venous catheter guide wire]. AB - A 74-year-old woman had a CVC inserted. Unfortunately, the guide wire was lost under the procedure, and despite a chest X-ray control it was not detected. The reason the guide wire was not identified was insufficient fine-tuning of the >>local<< PC monitor. We recommend regular fine-tuning of monitors used for X-ray control. PMID- 16221431 TI - [Pulmonary embolism complicated with right-sided intracardiac thrombus entrapped in a patent foramen ovale]. AB - With the increasing possibilities of and clinical demand for diagnostic imaging in the management of acute cardiovascular disease, the probability of uncovering complicated cases increases. These cases are often challenging to the clinician in determining the best method of treatment. We present a case of pulmonary embolism in which an intracardiac thrombus entrapped in a patent foramen ovale without penetrating it was discovered. The patient received thrombolytic therapy without complications and was discharged in good condition six days after thrombolysis. PMID- 16221433 TI - [Sleeping disorders can imitate attention deficit/hyperactivity disorder]. AB - Attention deficit/hyperactivity disorder (ADHD) is often associated with anxiety, depression, oppositional defiant disorder and sleeping difficulties. Sleeping disorders are reported in more than 50% of children with ADHD. On the other hand, symptoms of sleep disorder, inattention, hyperactivity and impulsiveness can imitate the symptoms of ADHD. We describe a 5(1/2)-year-old boy with symptoms of attention deficit and social withdrawal. His sleep was disturbed, with late sleep onset and frequent awakening during the night. After correction of his sleep pattern, the symptoms of attention deficit and social withdrawal disappeared. PMID- 16221434 TI - [The iron hormone hepcidin: A future diagnostic marker and therapeutic target in the treatment of anemia and hemochromatosis?]. PMID- 16221439 TI - [Psychologists can diagnose]. PMID- 16221449 TI - Arthrodesis of the first metatarsophalangeal joint for idiopathic hallux valgus: intermediate results. AB - BACKGROUND: Followup studies documenting the outcome of primary metatarsophalangeal (MTP) joint arthrodesis for treatment of hallux valgus deformities are rare. The purpose of this report was to evaluate the results of first MTP joint arthrodesis as treatment for moderate and severe hallux valgus deformities over a 22-year period in a single surgeon's practice. METHODS: All living patients treated between 1979 and 2001, for moderate and severe idiopathic hallux valgus deformities with first MTP joint arthrodesis were contacted and asked to return for a followup examination. Outcomes were assessed by comparing preoperative and postoperative pain, function, and radiographic appearance. First ray mobility and ligamentous laxity also were assessed postoperatively. RESULTS: Eighteen of 21 of the first MTP joints had successfully fused with the primary procedure at an average followup of 8.2 years (range 24 to 271 months). The time to union averaged 10 (range 7 to 15) weeks. Two of the three nonunions, both in the same patient, were asymptomatic and were not revised. One required a revision to achieve fusion. The average corrections in the hallux valgus angle and 1-2 intermetatarsal (IM) angle were 21 degrees and 6 degrees, respectively, and the average postoperative dorsiflexion angle was 22 degrees. Subjective satisfaction was rated as excellent in seventeen of 21 cases (80%) and good in the remaining four (20%). There was significant reduction in postoperative pain (p < 0.001), complete resolution of lateral metatarsalgia, and the postoperative American Orthopaedic Foot and Ankle Society (AOFAS) scores averaged 84 (range 72 to 90) at final followup. Major activity restrictions after surgery were uncommon, and all patients were able to wear conventional or comfort shoes. Interphalangeal (IP) joint arthritis progressed in seven of 21 feet (33%), but all of these changes were mild. CONCLUSIONS: In the present study, arthrodesis of the first MTP joint for idiopathic hallux valgus resulted in a high percentage of successful results at an average followup of over 8 years. PMID- 16221450 TI - Fresh ankle osteochondral allograft transplantation for tibiotalar joint arthritis. AB - BACKGROUND: Conventional treatment for tibiotalar joint arthritis relies on arthrodesis or prosthetic arthroplasty. Fresh osteochondral allografting is an alternative procedure to replace diseased articular cartilage. METHODS: Eleven patients (average age 43 years; range 18 to 65 years) had fresh osteochondral grafting of the tibiotalar joint. The diagnoses were posttraumatic arthritis in seven ankles, osteoarthritis in two, and an osteochondral defect in two. Precise cuts were made using the Agility (DePuy, Warsaw IN) ankle arthroplasty jigs. Bipolar replacements were used in nine ankles and unipolar in two. Results were evaluated using outcome scores, physical examinations, and standing ankle radiographs. RESULTS: At a minimum followup of 24 (average 33; range 26 to 45) months, six of the 11 ankles had successful grafting procedures. The average AOFAS score preoperatively improved from 55 to 73 postoperatively (p = 0.01). The patients' pain, gait, and walking surface scores were all significantly improved (p < 0.05). Of the five failures three underwent successful repeat allografting and one was revised to a total ankle arthroplasty, and one has had no further surgery. The ankle range of motion arc was 30 degrees or more in six ankles. Additional surgery included five talofibular joint debridements, three repeat graftings, two hardware removals, and one conversion to a prosthetic ankle replacement. There was one intraoperative fibular fracture and one superficial wound infection. The serum of 10 patients tested positive for cytotoxic HLA antibodies postoperatively. Radiographs revealed moderate and severe joint degeneration in six ankles; however, this did not necessarily correlate with a poor outcome. Poor results tended to occur in ankles with a graft-host size mismatch or graft thickness of less than 7 mm. CONCLUSION: Fresh osteochondral transplantation for tibiotalar joint arthritis is a promising alternative to arthrodesis and prosthetic replacement. Early results demonstrate successful outcomes and good pain relief in over half the patients in this series. PMID- 16221451 TI - Evaluation of hallux alignment and functional outcome after isolated tibial sesamoidectomy. AB - BACKGROUND: Functional loss and clinical evidence of hallux malalignment have been reported to follow isolated tibial sesamoidectomy. METHODS: Thirty-two patients with isolated tibial sesamoidectomies were identified. Patients with a diagnosis of peripheral neuropathy, diabetes mellitus, inflammatory arthropathy or previous foot surgery were excluded as were patients who had concomitant joint realignment procedures. Twenty patients were available for followup with the Short Form-36 (SF-36), Foot Function Index (FFI) disability scale, visual analog scale (VAS), and questionnaire at an average of 62 (range 10 to 157) months after surgery. Fourteen patients returned for physical examination, radiographs, and pedographic and isokinetic examination. RESULTS: Physical examination of the 14 patients did not reveal any significant change in clinical alignment, range of motion or tenderness. Preoperative and postoperative comparison radiographs did not reveal significant differences in the intermetatarsal (IM) angle, hallux valgus (HV) angle distal metatarsal articular angle (DMAA), or sesamoid alignment (sesamoid station). Postoperative outcome measurements (VAS, SF36, and FFI) for 20 patients found significant relief of pain and improved functional outcome. Computerized dynamic pedographic measurements (Performance Orthotic) for 12 patients did not reveal any altered plantar pressures in the region of the hallux metatarsophalangeal joint. Isokinetic measurements of ankle plantar flexion push off strength in eight patients did not reveal significant differences in side-to side measurements. Eighteen of 20 (90%) patients indicated that they were able to resume all preoperative activities; six (30%) had extreme difficulty or an inability to stand on tip toe, but this did not impact their activities of daily living or their athletic endeavors. Two patients (14.3%) developed transfer metatarsalgia, but only one was symptomatic. CONCLUSION: Isolated tibial sesamoidectomy is a safe and effective treatment for recalcitrant tibial sesamoiditis. Hallux malalignment and deformity resulting in functional loss and change in hallux alignment can be avoided by meticulous surgical technique with repair of the soft tissues. PMID- 16221452 TI - Tibiotalocalcaneal (hindfoot) arthrodesis by retrograde intramedullary nailing using a curved locking nail. The results of 52 procedures. AB - BACKGROUND: The purpose of this study was to report the results of 52 combined subtalar and ankle arthrodesis using an intramedullary nail. METHODS: Retrospective review identified 49 patients who had 52 combined ankle and subtalar arthrodeses with an ACE retrograde locked intramedullary humeral nail (DePuy-Ace), Warsaw, IN). Most procedures included bone grafts from the fibula, proximal tibia, or iliac crest or femoral head allograft. Intraoperative complications included one fractured tibia and one fractured medial malleolus. The procedure was done mainly for the treatment of combined ankle and subtalar arthritis (31) or complex hindfoot deformities (12). Outcome was assessed by a combination of chart review, clinical examination, and telephone questionnaire. Followup averaged 34 (8 to 73) months. RESULTS: At followup 82% of patients were satisfied with the results of surgery, 82% reported improvements in pain levels, and 67% reported improved foot function. The average postoperative American Orthopaedic Foot and Ankle Society (AOFAS) hindfoot score was 63. Postoperative complications included deep infection, amputation, stress fracture, nonunion, and prominent hardware. CONCLUSION: Hindfoot arthrodesis with intramedullary nailing is an effective technique for treating complex foot deformities and often is the only alternative to amputation. Patient satisfaction is high, but the procedure is demanding and complications are frequent. PMID- 16221453 TI - An analysis of outcome measures following the Brostrom-Gould procedure for chronic lateral ankle instability. AB - BACKGROUND: The Brostrom-Gould procedure is a commonly recommended operative treatment for chronic ankle instability. Using standardized physician-based outcome scores, the results of this procedure have been uniformly excellent. Current scoring systems, however, do not adequately evaluate mechanical or functional instability. Therefore, outcome data may suggest greater success than is justified. METHODS: A retrospective review was done of 73 patients who had isolated Brostrom-Gould repairs of the lateral ankle ligaments. The mean time to followup was 64 months. Both the AOFAS ankle-hindfoot score and the Short Form 36 (SF-36) were used to evaluate outcome. RESULTS: The overall American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score was 95 of 100 points. Despite that, 17% of patients in the study had functional instability of the ankle that was not reflected in the AOFAS score. The mean physical component score of the SF 36 was 84% and reflected the presence of functional instability. Low correlations were found between the AOFAS ankle-hindfoot score and the SF-36 score. CONCLUSIONS: The current study identified a deficiency in the AOFAS score in evaluating functional ankle stability after the Brostrom-Gould procedure. A more meaningful analysis of outcomes can be expected using the SF-36 score. The data suggest that greater attention must be paid to functional rehabilitation after ankle stabilization surgery to obtain optimal outcome. PMID- 16221454 TI - Radiographic assessment of adult flatfoot. AB - BACKGROUND: The accurate measurement of flatfoot on standing radiographs allows correct diagnosis of the condition and evaluation of reconstructive procedures. METHOD: The standing radiographic measurements of patients with symptomatic, adult flatfoot were compared to controls using blinded observers. RESULTS: On the lateral radiograph, the talar-to-first metatarsal angle, the calcaneal pitch angle, and the medial cuneiform-fifth metatarsal height differed significantly between the patient group and the controls. The difference in the talar-to-first metatarsal angles on lateral radiographs was the most statistically significant (patient group 21.1 +/- 10.8 degrees and control 7.1 +/- 10.7 degrees, p < 0.0001) with good correlation between readings (intraobserver 0.75, interobserver 0.83). On the anteroposterior (AP) radiograph, the talar head uncoverage distance was the most significantly different measurement between these groups. CONCLUSIONS: These findings support the hypothesis that the talar-first metatarsal angle is an accurate radiographic identifier of patients with symptomatic, adult flatfoot. PMID- 16221455 TI - Treatment of rupture of the Achilles tendon with fibrin sealant. AB - BACKGROUND: The optimal management strategy for acute Achilles tendon ruptures is controversial. These injuries historically were treated by nonoperative methods (cast immobilization, bandaging); however, operative repair of the ruptured tendon has become popular. METHODS: Thirty-two patients who had rupture of the Achilles tendon were treated operatively with use of fibrin sealant, and clinical and functional performance measures were assessed after a mean followup of at least 6 months between November, 1998, and July, 2003. All of the patients were male. Average age was 38.18 (30 to 45) years. All of the patients were followed for at least 18 months after surgery. Average followup time was 22.4 (18 to 56) months. We evaluated all patients according to the scoring system of Thermann et al. RESULTS: Our results were excellent in 24 patients and good in eight patients. One patient had rerupture 3 weeks after surgery. CONCLUSION: Fibrin sealants are biologically compatible, hemostatic agents derived from human plasma that can be used instead of suture or suture support. We think that the treatment of rupture of the Achilles tendon with fibrin sealant is a useful treatment, and there is less risk of complications, such as deep infection, than in other operative procedures. We had no wound closure problems, the incision size was small, and the operating time was short. However, it must be remembered that the risk of disease transfer by fibrin sealant application is still present. PMID- 16221456 TI - Exogenously administered substance P and neutral endopeptidase inhibitors stimulate fibroblast proliferation, angiogenesis and collagen organization during Achilles tendon healing. AB - BACKGROUND: In the last few years much research has been conducted in methods to promote tendon healing. The aim of this study was to determine if the healing process after operative repair of rat Achilles tendons could be stimulated by the paratendinous injection of a sensory peptide, substance P (SP). METHODS: Ninety six male Sprague-Dawley rats were randomly allocated to four groups: (I) control buffer injections, (II) injections of SP 10(-6) mol/kg body weight combined with a carrier, (III) injections of SP 10(-8) mol/kg BW with the carrier, and (IV) injections with the carrier only (thiorphan 1 micromol/kg BW and captopril 30 micromol/kg BW, both neutral endopeptidase inhibitors). The influence on tissue repair was determined from the histologic measurement of fibroblast proliferation, angiogenesis, and collagen organization. On days 7, 14, 28 and 42, animals were sacrificed and histologic evaluations were performed on the injured Achilles tendon constructs. RESULTS: The two groups subjected to SP injections showed a significant initial fibroblast proliferation on day 7 (p < 0.05), which rapidly declined by day 14 to the level of cellular proliferation observed with the use of thiorphan and captopril. Capillary proliferation showed a similar evolution, except that in the second week angiogenesis in the treated groups was below the level of the control group. Strikingly, collagen orientation increased faster in the groups injected with SP. This was obvious from the second week already and the difference remained until the completion of the study. CONCLUSION: This is the first study to demonstrate that paratendinous injections of SP after operative repair of the Achilles tendon in rats appears to provide a boost to the initial stages of healing and significantly accelerate the reparative phase of the healing process. PMID- 16221457 TI - The use of autologous concentrated growth factors to promote syndesmosis fusion in the Agility total ankle replacement. A preliminary study. AB - BACKGROUND: The Agility (DePuy, Warsaw, Indiana) total ankle replacement has been in use since 1984. One of the most common complications continues to be delayed union or nonunions of the distal tibiofibular syndesmosis. In the reported studies on the Agility ankle the delayed union and nonunion rate can be as high as 38%. METHODS: Since 1999, 114 Agility total ankle replacements were done at two centers in the United States without the use of autologous concentrated growth factors. Since July of 2001, 66 Agility ankles were implanted with Symphony (DePuy, Warsaw, Indiana) augmented bone grafting. The standard operative technique was followed in all the patients. Prospective data was collected on all patients. The standard ankle radiographs were taken preoperatively and postoperative at 8 weeks, 12 weeks, 16 weeks, 6 months, and yearly. CT scans were obtained at 6 months if fusion at the syndesmosis was questionable. The Graphpad Instat software (Graphpad Software Inc., San Diego, CA) was used for statistical analysis. The two-tailed unpaired t-test was used, and the value <0.05 was considered significant. RESULTS: There was no statistical difference in the demographic data for the two groups. In 114 ankle replacements without autologous concentrated growth factors 70 fused at 8 weeks (61%), 14 fused at 12 weeks (12%), 13 fused at 6 months (12%). There were 17 nonunions (15%); delayed unions (3 to 6 months) and nonunions, therefore, equaled 27%. The syndesmosis fused in 50 of the 66 ankle replacements (76%) that had autologous concentrated growth fractures at 8 weeks (76%); 12 fused at 3 months (18%), 2 fused at 6 months (3%), 2 had nonunions (3%). Delayed unions (3 to 6 months) and nonunions equaled 6%. There was a statistically significant improvement in the 8- and 12-week fusion rates, and a statistically significant reduction in delayed unions and nonunions. CONCLUSION: Autologous concentrated growth factors appear to make a significant positive difference in the syndesmosis union rate in total ankle replacements. PMID- 16221458 TI - Assessing foot temperature using infrared thermography. AB - BACKGROUND: Previous reports recommended using skin temperature as a guide to monitor neuropathic feet during their rehabilitation course. However, the diagnostic usefulness was limited because of poor thermal measurement and procedures. The purpose of this study was to propose a standardized protocol to quantify foot temperature. METHODS: An infrared image system was used to measure skin temperature. The first experiment was conducted on 16 healthy volunteers to study temperature variation with respect to time. This study mapped out six subregions of anatomic interest over the sole, and average temperature values for each were studied. The second experiment was conducted on 62 diabetic patients, with and without sympathetic skin response (SSR), to study proposed sole temperature normalization with respect to forehead temperature for clinical diagnosis. RESULTS: In the first experiment, the temperature in each plantar subregion varied as a function of time. In the sole area, the highest temperature was noted in the arch region (29.3 +/- 0.9 degrees C). The toes had the lowest temperature value (26.2 +/- 1.2 degrees C) in all areas. Equilibrium was reached after 15 minutes for the mean plantar temperature (27.8 +/- 1.0 degrees C). In the second experiment, the diabetic patients without SSR had a slightly higher mean plantar temperature (27.6 +/- 1.8 degrees C) than those with SSR (26.8 +/- 2.2 degrees C), but the difference was not statistically significant (p > 0.05). The SSR-absent group (0.19) and the SSR-present group (0.24) had significant differences in their normalized temperatures as proposed (p < 0.05). CONCLUSIONS: The mean temperature of the entire plantar area was found to be more stable than the individual subregions, serving as a more practical indicator for thermoregulatory functions. The study also found that the overall mean plantar temperature stabilized after 15 minutes, and, thus, this time was recommended for clinical thermographic measurements. The normalized temperature may have more useful application than the plantar absolute temperature, as exemplified by the better correlation in diabetic feet. The mean plantar temperature, the wait time to start measurement, and the proposed normalization are believed to play important roles in neuropathic foot disorders. PMID- 16221459 TI - Plantar-to-dorsal compared to dorsal-to-plantar screw fixation for proximal chevron osteotomy: a biomechanical analysis. AB - BACKGROUND: A change in screw orientation in fixing the chevron proximal first metatarsal osteotomy was noted anecdotally to improve fixation strength. The authors hypothesized that plantar-to-dorsal screw orientation would be more stable than the conventional dorsal-to-plantar screw orientation for fixation of the chevron osteotomy. The purpose of this study was to determine if the load-to failure and stiffness of the chevron type proximal first metatarsal osteotomy stabilized using plantar-to-dorsal screw fixation were greater than with the more conventional dorsal-to-plantar screw fixation method. METHODS: One foot from each of eight matched cadaver pairs was randomly assigned to one of two groups: 1) fixation with a dorsal-to-plantar lag screw or 2) fixation with a plantar-to dorsal lag screw. A proximal chevron osteotomy was then created using standard technique and the metatarsal was fixed according to previously established method. The bone was potted in polyester resin, and the construct was fitted into a materials testing system machine in which load was applied to the plantar aspect of the metatarsal until failure. The two groups were compared using a two tailed Student t test. RESULTS: The average load-to-failure and stiffness of the chevron osteotomy fixed with the plantar-to-dorsal lag screw were significantly greater (p < 0.05) than the group fixed with more conventional dorsal-to-plantar lag screws. CONCLUSION: Plantar-to-dorsal screw orientation was more stable than the conventional dorsal-to-plantar screw orientation for fixation of the proximal chevron osteotomy. Plantar-to-dorsal screw orientation should be considered when using the chevron proximal first metatarsal osteotomy. PMID- 16221460 TI - The effect of terrain on foot pressures during walking. AB - BACKGROUND: High plantar pressures are associated with the development of foot ulcers in people with diabetic neuropathy. The effect of terrain on plantar pressures during walking has not been fully explored. METHODS: Twenty 23- to 40 year-old subjects with no known musculoskeletal pathology walked across three terrains: padded carpet (R), grass (G), and concrete (C) while wearing Novel Pedar (Novel Electronics Inc. GMBH, Munich, Germany) insoles with and without shoes. Pressures were collected at 50 Hz. The sole of the foot was divided into: heel (H), lateral midfoot (LM), medial midfoot (MM), big toe and first metatarsal head (BT), and lateral toes and metatarsal heads (LT). Repeated measures ANOVA identified differences in pressures, forces, and contact areas across terrains. Post hoc Bonferroni adjustments were used to accept an overall alpha level of 0.05. RESULTS: Peak pressure (PP), maximal mean pressure (MMP) and pressure time integral (PTI) were significantly higher (p < 0.01) when walking barefoot on concrete than on grass or carpet for all foot regions except MM and LM. The percent increase in the three pressure variables ranged from 21% to 43%. Grass and carpet PP, MMP, and PTI were similar for the BT and LT. Wearing shoes significantly increased contact area and decreased all pressure variables on all three terrains. CONCLUSION: Walking barefoot, especially on concrete was associated with higher plantar pressure variables. Wearing shoes eliminated terrain differences in pressure except under the lesser toes. Persons with insensate feet should avoid walking barefoot on hard surfaces to avoid excessive plantar pressures. Wearing shoes and covering hard floors with nonslip, padded rugs may decrease plantar pressures and the risk of ulceration. PMID- 16221461 TI - Joint stabilizing response to expected and unexpected tilts. AB - BACKGROUND: The results found in the literature regarding functional ankle joint stabilization are controversial possibly because of the testing methods used. METHODS: The responses of 22 subjects to unexpected and expected sudden inversions of the foot were compared for all subjects together, as well as grouped by their self-perceived stability. A three-dimensional foot model was used to describe ankle and foot motion. Electromyographic signals of six muscles of the lower limb as well as the horizontal ground reaction forces were recorded. RESULTS: Whereas unexpected and expected trials did not show significant differences (p > 0.05) in kinematics, higher activation and horizontal force integrals were found for the unexpected trials. In addition, no differences in electromyographic or ground reaction force parameters were found between stable and unstable ankles; however, the kinematics revealed higher amplitudes and velocities for the stable group. CONCLUSIONS: The awareness of the instant of tilt enhances stabilization in that the same motion is achieved with a lower muscle activation. Evidence suggests that this is triggered at supraspinal levels. We found that timing of the EMG signal is not as relevant to stabilization as the amplitude (which has often been disregarded in the literature). PMID- 16221462 TI - A metatarsal equivalent to the metacarpal index in Marfan syndrome. AB - BACKGROUND: Foot problems are common in patients with Marfan syndrome because of the ligamentous laxity that affects the weightbearing joints most. Such patients frequently are seen by their general practitioners or podiatrists. Educating primary health care providers about a metatarsal index, if it is sufficiently sensitive and specific, may help patients get early and appropriate workup for connective tissue disorders. METHODS: A metatarsal equivalent to the metacarpal index (MCI) in the hand was evaluated as a diagnostic tool for Marfan syndrome (and possibly other connective tissue disorders). Fifty-six patients were studied. Sixteen had a genetic diagnosis of Marfan syndrome. There were 20 controls each for the MCI and the metatarsal index (MTI). Hand and foot radiographs were reviewed. RESULTS: The average MCI in patients with Marfan syndrome was 9.8 compared to 7.6 in the control group (p < 0.0005). The average MTI was 12.7 and 9.8, respectively (p < 0.0005). An MCI value of 8.5 and an MTI value of 10.5 had the best statistical profiles (combination of sensitivity and specificity) in diagnosing Marfan syndrome in our study population. CONCLUSION: The MTI as a screening tool for Marfan syndrome is as effective as, if not more than, the well-recognized MCI. PMID- 16221463 TI - Bilateral hallux valgus associated with os intermetatarseum: a case report. PMID- 16221464 TI - The peroneocalcaneus internus muscle: an unusual cause of posterior ankle impingement. PMID- 16221465 TI - Bilateral os trigonum syndrome associated with bilateral tenosynovitis of the flexor hallucis longus muscle. PMID- 16221466 TI - Clinical tip: postoperative dressing for hallux valgus surgery. PMID- 16221467 TI - Clinical tip: modified Akin osteotomy. PMID- 16221468 TI - Estimating crashes involving heavy vehicles in Western Australia, 1999-2000: a capture-recapture method. AB - A two-sample exploratory study of police and hospital records was undertaken to estimate the number of fatalities and serious injuries for heavy vehicle drivers involved in a crash in Western Australia. The capture-recapture method was used to assess differences and similarities in characteristics of heavy vehicle drivers from both sources. Each heavy vehicle driver involved in a crash from the police report was matched against the heavy vehicle driver's hospitalisation record from the Hospital Morbidity Data System, with surname, initials, date of birth, gender, date of crash, road user type and vehicle type as matching fields. The estimated number of fatalities and serious injuries to heavy vehicle drivers from 1st July 1999 to 31st December 2000 was 5 and 59, respectively, which was 25 and 31% higher based on the capture-recapture methodology than the aggregated (non-overlapping) total officially reported to the police and hospitals. No significant age difference (p>0.05) was found for drivers involved in a heavy vehicle crash between the two sources (37 years versus 40 year of age). However, female heavy vehicle drivers were over-represented in the hospital records (11%) compared to the police records (1%). The capture-recapture approach is useful for evaluating the completeness of data sources and identifying biases within datasets. The underestimation of heavy vehicle drivers seriously injured and killed has important implications for heavy vehicle safety management and resource allocation in Western Australia. PMID- 16221469 TI - Boldine and its antioxidant or health-promoting properties. AB - The increasing recognition of the participation of free radical-mediated oxidative events in the initiation and/or progression of cardiovascular, tumoural, inflammatory and neurodegenerative disorders, has given rise to the search for new antioxidant molecules. An important source of such molecules has been plants for which there is an ethno-cultural base for health promotion. An important example of this is boldo (Peumus boldus Mol.), a chilean tree whose leaves have been traditionally employed in folk medicine and is now widely recognized as a herbal remedy by a number of pharmacopoeias. Boldo leaves are rich in several aporphine-like alkaloids, of which boldine is the most abundant one. Research conducted during the early 1990s led to the discovery that boldine is one of the most potent natural antioxidants. Prompted by the latter, a large and increasing number of studies emerged, which have focused on characterizing some of the pharmacological properties that may arise from the free radical scavenging properties of boldine. The present review attempts to exhaustively cover and discuss such studies, placing particular attention on research conducted during the last decade. Mechanistic aspects and structure-activity data are discussed. The review encompasses pharmacological actions, which arise from its antioxidant properties (e.g., cyto-protective, anti-tumour promoting, anti inflammatory, anti-diabetic and anti-atherogenic actions), as well as those that do not seem to be associated with such activity (e.g., vasorelaxing, anti trypanocidal, immuno- and neuro-modulator, cholagogic and/or choleretic actions). Based on the pharmacological and toxicological data now available, further research needs and recommendations are suggested to define the actual potential of boldine for its use in humans. PMID- 16221470 TI - Urinary cadmium and mortality among inhabitants of a cadmium-polluted area in Japan. AB - The influence of cadmium (Cd) body burden on mortality remains controversial. Excess mortality and the dose-response relationship between mortality and urinary cadmium excretion were investigated in this study among environmentally exposed subjects. A 15-year follow-up study was carried out on 3119 inhabitants (1403 men and 1716 women) of the Cd-polluted Kakehashi River basin, whose urinary Cd concentration was examined in a 1981-1982 health impact survey. The mortality risk of high urinary Cd (> or = 10 microg/g Cr) subjects after adjustment for age using Cox's proportional hazard model was higher than that of moderate urinary Cd (< 10 microg/g Cr) subjects in both sexes. When the subjects were divided into five groups according to the amount of urinary Cd (<3, 3-5, 5-10, 10-20, > or = 20 microg/g Cr), the mortality risk was significantly increased among the subjects with urinary Cd > or = 3 microg/g Cr in proportion to the increases in the amount of urinary Cd concentration after adjustment for age, especially in women. Furthermore, special causes of death among high and moderate urinary Cd were investigated, and mortality risk ratio for heart failure, which is a cause of death often diagnosed in cases with a gradual deterioration culminating in death, was significantly increased in both sexes, compared with the moderate urinary Cd subjects. Also, in women the mortality risk for renal diseases in the high urinary Cd subjects was significantly higher than that in the moderate urinary Cd subjects. These results suggest that a causal association between Cd body burden and mortality exists among inhabitants environmentally exposed to Cd but that no special disease may be induced except renal diseases. PMID- 16221471 TI - Time trends in burdens of cadmium, lead, and mercury in the population of northern Sweden. AB - The time trends of exposure to heavy metals are not adequately known. This is a worldwide problem with regard to the basis for preventive actions and evaluation of their effects. This study addresses time trends for the three toxic elements cadmium (Cd), mercury (Hg), and lead (Pb). Concentrations in erythrocytes (Ery) were determined in a subsample of the population-based MONICA surveys from 1990, 1994, and 1999 in a total of 600 men and women aged 25-74 years. The study took place in the two northernmost counties in Sweden. To assess the effect of changes in the environment, adjustments were made for life-style factors that are determinants of exposure. Annual decreases of 5-6% were seen for Ery-Pb levels (adjusted for age and changes in alcohol intake) and Ery-Hg levels (adjusted for age and changes in fish intake). Ery-Cd levels (adjusted for age) showed a similar significant decrease in smoking men. It is concluded that for Pb and maybe also Hg the actions against pollution during recent decades have caused a rapid decrease of exposure; for Hg the decreased use of dental amalgam may also have had an influence. For Cd, the decline in Ery-Cd was seen only in smokers, indicating that Cd exposure from tobacco has decreased, while other environmental sources of Cd have not changed significantly. To further improve the health status in Sweden, it is important to decrease the pollution of Cd, and actions against smoking in the community are important. PMID- 16221472 TI - Heat acclimatization and sunshine cause false indications of mortality due to ozone. AB - We used graphic analysis and generalized additive modeling to assess whether mortality that is often attributed to ozone and sometimes to < 10-microm particulate matter (PM10) and sulfur dioxide in hot weather results from confounding by neglected weather factors. When mean daily air temperatures exceeded 18 degrees C, mortality at age > or = 65 years, in Greater London 1991 2002 rose and at each temperature was generally higher in early summer than later after exposure to hot weather. Ozone, and to lesser degrees PM10 and sulfur dioxide, followed similar patterns. At each temperature, high levels of ozone and PM10 tended to be associated with sunshine, and high PM10 and sulfur dioxide with low wind, both of which will increase heat stress. With allowance for these confounding factors, generalized additive modeling showed no significant mortality due to ozone, to PM10, or to sulfur dioxide. We conclude that the pollutants played little part in excess mortality associated with hot weather. PMID- 16221473 TI - Alteration of host cell phenotype by Theileria annulata and Theileria parva: mining for manipulators in the parasite genomes. AB - The apicomplexan parasites Theileria annulata and Theileria parva cause severe lymphoproliferative disorders in cattle. Disease pathogenesis is linked to the ability of the parasite to transform the infected host cell (leukocyte) and induce uncontrolled proliferation. It is known that transformation involves parasite dependent perturbation of leukocyte signal transduction pathways that regulate apoptosis, division and gene expression, and there is evidence for the translocation of Theileria DNA binding proteins to the host cell nucleus. However, the parasite factors responsible for the inhibition of host cell apoptosis, or induction of host cell proliferation are unknown. The recent derivation of the complete genome sequence for both T. annulata and T. parva has provided a wealth of information that can be searched to identify molecules with the potential to subvert host cell regulatory pathways. This review summarizes current knowledge of the mechanisms used by Theileria parasites to transform the host cell, and highlights recent work that has mined the Theileria genomes to identify candidate manipulators of host cell phenotype. PMID- 16221474 TI - The L162V polymorphism at the peroxisome proliferator activated receptor alpha locus modulates the risk of cardiovascular events associated with insulin resistance and diabetes mellitus: the Veterans Affairs HDL Intervention Trial (VA HIT). AB - BACKGROUND: The Veterans Affairs HDL Intervention Trial (VA-HIT) showed that gemfibrozil, which activates peroxisome proliferator-activator receptor alpha (PPARalpha), significantly reduced the risk of cardiovascular (CV) events in men with low HDL cholesterol (< 40 mg/dl) and established coronary heart disease. Treatment was particularly beneficial in those with insulin resistance (IR) or diabetes mellitus (DM). We hypothesized that the association between a functional polymorphism at the PPARA locus (L162V) and the risk of a CV event, as well as response to fibrate therapy, might be greatest in those with either IR or DM (DM/IR) in VA-HIT. METHODS AND RESULTS: A total of 827 men (placebo, n = 413; gemfibrozil, n = 414) from the VA-HIT were genotyped. This population included a high proportion of subjects with DM/IR. In VA-HIT, the PPARA V162 allele was associated with reduced levels of HDL cholesterol and the presence of DM/IR at baseline. It was also associated with reduced risk of CV events in those with DM/IR but not in those with neither (DM/IR *PPARA genotype, P = 0.005). Among subjects with DM/IR, treatment with gemfibrozil reduced CV events in non-carriers from 29.9 to 17.8% and carriers of the V162 allele from 14.7 to 4.8%. In contrast, carriers of the V162 allele with no DM/IR who were treated with gemfibrozil experienced significantly more CV events than did those who received placebo (20.6% versus 13.6%; P = 0.01). CONCLUSIONS: The effect of the L162V polymorphism at the PPARA locus on CV risk depends on the presence of DM/IR. Among subjects treated with gemfibrozil, the V162 allele was associated not only with reduced CV risk in subjects with DM/IR, but also with significantly increased CV risk in the absence of these traits, identifying this genetic variant as a potential marker for predicting which subjects may have a favorable response to fibrate therapy. PMID- 16221475 TI - Computational analysis of type II endoleaks in a stented abdominal aortic aneurysm model. AB - Insertion of a stent-graft into an aneurysm to form a new (synthetic) blood vessel and prevent the weakened artery wall from rupture is an attractive surgical intervention when compared to traditional open surgery. However, focusing on a stented abdominal aortic aneurysm (AAA), post-operative complications such as endoleaks may occur. An endoleak is the net influx of blood during the cardiac cycle into the cavity (or sac) formed by the stent-graft and the AAA wall. A natural endoleak source may stem from one or two secondary branches leading to and from the aneurysm, labeled types IIa and IIb endoleaks. Employing experimentally validated fluid-structure interaction solvers, the transient 3-D lumen and cavity blood flows, wall movements, pressure variations, maximum wall stresses and migration forces were computed for types IIa and IIb endoleaks. Simulation results indicate that the sac pressure caused by these endoleaks depends largely on the inlet branch pressure, where the branch inlet pressure increases, the sac pressure may reach the systemic level and AAA-rupture is possible. The maximum wall stress is typically located near the anterior distal side in this model, while the maximum stent-graft stress occurs near the bifurcating point, in both cases, due to local stress concentrations. The time varying leakage rate depends on the pressure difference between AAA sac and inlet branch. In contrast, the stent-graft migration force is reduced by type II endoleaks because it greatly depends on the pressure difference between the stent graft and the aneurysm cavity. PMID- 16221476 TI - Selective determination of gamma-aminobutyric acid, glutamate and alanine by mixed micellar electrokinetic chromatography and fluorescence detection. AB - A mixed micellar electrokinetic chromatography method with fluorescence detection was developed to simultaneously monitor gamma-aminobutyric acid (GABA), glutamate (Glu) and alanine (Ala) in biological samples. Amino acids were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA). The separation of three NDA-labeled isomers (GABA, alpha-ABA, beta-ABA) was studied in detail with different micelles solutions such as sodium dodecyl sulfate (SDS), beta-cyclodextrin (beta-CD) and sodium cholate (SC). Simultaneous resolution of GABA, Glu and Ala from 21 amino acids was achieved within 5 min using 20 mM phosphate buffer at pH 8.7 containing 24 mM SC and 26 mM SDS. The detection limits were 4.0 x 10(-8), 1.1 x 10(-8) and 1.3 x 10(-8) M, for GABA, Glu and Ala, respectively, with S/N = 2. The method was applied to monitor the changes of amount of GABA, Glu and Ala in tobacco leaf in response to cold and dark stress. PMID- 16221478 TI - Theory of home range estimation from displacement measurements of animal populations. AB - A theory is provided for the estimation of home ranges of animals from displacement measurement procedures. The theoretical tool used is the Fokker Planck equation, its characteristic quantities being the diffusion constant which describes the motion of the animals, and the attractive potential which addresses their tendency to live in restricted regions, e.g., near their burrows. The measurement technique is shown to correspond to the calculation of a certain kind of mean square displacement of the animals relevant to the specific probing window in space corresponding to the region of observation. The output of the theory is a sigmoid curve of the observable mean square displacement as a function of the ratio of distances characteristic of the home range and the measurement window, along with an explicit prescription to extract the home range from observations. Applications of the theory to rodent movement in Panama and New Mexico are pointed out. An analysis is given of the sensitivity of our theory to the choice of the confining potential via the use of various representative cases. A comparison is provided between home range size inferred from our method and from other procedures employed in the literature. Consequences of home range overlap are also discussed. PMID- 16221477 TI - Liquid chromatography/tandem mass spectrometric confirmatory method for determining aflatoxin M1 in cow milk: comparison between electrospray and atmospheric pressure photoionization sources. AB - A liquid chromatography/electrospray (ESI)-tandem mass spectrometric method for the measurement of aflatoxin M1 (AFM1) in milk is described. Milk sample after protein precipitation with acetone was cleaned-up with a Carbograph-4 cartridge. Performances of the ESI source were compared with those of the atmospheric pressure photoionization source (APPI). Although a method quantification limit (MQL) of 6 ng/kg could be achieved operating with APPI source with respect to an MQL of 12 ng/kg with ESI, all the other performances being similar, then ESI was preferred as being more robust and widespread at present. PMID- 16221479 TI - Additional morphological and physiological heterogeneity within the midgut of larval Aedes aegypti (Diptera: Culicidae) revealed by histology, electrophysiology, and effects of Bacillus thuringiensis endotoxin. AB - Analysis of larval Aedes aegypti midgut using scanning electron microscopy, nuclear and mitochondrial dyes, response to Bacillus thuringiensis israelensis CryIVB toxin, and electrophysiology is described. The anterior ventriculus ("stomach") region is found to have much lower mitochondrial densities than other midgut regions. The transitional region is distinguished by apical surface architecture, and by region-specific effects of CryIVB endotoxin. In this region CryIVB causes holes ranging from 1.0 to 7.0 microm in diameter (mean 3.3+/-0.53 microm, N=12), blisters 16.9+/-1.54 microm in diameter (N=10), and separation of adjacent cells. The holes are not consistent with damage due to the colloid osmotic lysis model of delta-endotoxin activity. The posterior ventriculus possesses a distinctive cellular architecture consisting of hemispherical, domed apical membranes surrounded by deep clefts. Functional and morphological heterogeneity is revealed within the posterior ventriculus, with the anterior end dominating the electrical profile of isolated, perfused preparations and showing the greatest response to serotonin. Hyperpolarization of the transepithelial potential by serotonin occurred in conjunction with a decrease in the space constant lambda, ruling out closure of ion channels as the mechanism of action of serotonin. PMID- 16221480 TI - Phosphorus retention and sorption by constructed wetland soils in Southeast Ireland. AB - It may be necessary to use constructed wetlands as a land use practice to mitigate phosphorus (P) loss from agriculture in Ireland. The objectives of this study were to determine the ability of two constructed wetland site soils to retain and sorb P. Intact soil/water column studies were used to determine P release/retention rates during a 30-day incubation period. Soil columns flooded with distilled water released P during the first 2 days; however, soluble reactive P (SRP) concentrations in overlying floodwaters decreased thereafter. Soils with overlying floodwaters spiked at 5 and 15 mg SRP L(-1) retained highest amounts of P (p < 0.05) with retention at these concentrations controlled by SRP in overlying waters. Retention rates by soils ranged between 0.3 and 60.9 mg Pm( 2) d(-1). Maximum P sorption capacity (Smax) was higher for wetland soils at Dunhill, Waterford (1464 mg P kg(-1)) in comparison to soils at Johnstown Castle, Wexford (618 mg P kg(-1)). Equilibrium P concentrations (EPC0) were low (in the microg SRP L(-1) range), indicating a high capacity of these soils to sorb P. Phosphorus sorption parameters were significantly related to ammonium oxalate extractable aluminium (Al) and iron (Fe) content of soils. PMID- 16221481 TI - Peracetic acid (PAA) disinfection of primary, secondary and tertiary treated municipal wastewaters. AB - The efficiency of peracetic acid (PAA) disinfection against enteric bacteria and viruses in municipal wastewaters was studied in pilot-scale. Disinfection pilot plant was fed with the primary or secondary effluent of Kuopio municipal wastewater treatment plant or tertiary effluent from the pilot-scale dissolved air flotation (DAF) unit. Disinfectant doses ranged from 2 to 7 mg/l PAA in the secondary and tertiary effluents, and from 5 to 15 mg/l PAA in the primary effluents. Disinfection contact times were 4-27 min. Disinfection of secondary and tertiary effluents with 2-7 mg/l PAA and 27 min contact time achieved around 3 log reductions of total coliforms (TC) and enterococci (EC). PAA disinfection also significantly improved the hygienic quality of the primary effluents: 10-15 mg/l PAA achieved 3-4 log reductions of TC and EC, 5 mg/l PAA resulting in below 2 log reductions. F-RNA coliphages were more resistant against the PAA disinfection and around 1 log reductions of these enteric viruses were typically achieved in the disinfection treatments of the primary, secondary and tertiary effluents. Most of the microbial reductions occurred during the first 4-18 min of contact time, depending on the PAA dose and microorganism. The PAA disinfection efficiency remained relatively constant in the secondary and tertiary effluents, despite of small changes of wastewater quality (COD, SS, turbidity, 253.7 nm transmittance) or temperature. The disinfection efficiency clearly decreased in the primary effluents with substantially higher microbial, organic matter and suspended solids concentrations. The results demonstrated that PAA could be a good alternative disinfection method for elimination of enteric microbes from different wastewaters. PMID- 16221482 TI - Conventional and thermophilic aerobic treatability of high strength oily pet food wastewater using membrane-coupled bioreactors. AB - Although thermophilic treatment systems have recently gained considerable interest, limited information exists on the comparative performances of membrane coupled bioreactors (MBR) at thermophilic and conventional conditions. In this study aerobic MBRs operating at room temperature (20 degrees C) and at lower thermophilic range (45 degrees C) were investigated for the treatment of dissolved air flotation (DAF) pretreated pet food wastewater. The particular wastewater is characterized by oil and grease (O & G) concentrations as high as 6 g/L, COD of 51 g/L, BOD of 16 g/L and volatile fatty acid (VFA) of 8.3 g/L. The performances of the two systems in terms of COD, BOD and O & G removal at varying hydraulic retention time (HRT) are compared. COD removal efficiencies in the thermophilic MBR varied from 75% to 98% and remained constant at 94% in the conventional MBR. The O & G removal efficiencies were 66-86% and 98% in the thermophilic and conventional MBR, respectively. Interestingly, high concentrations of VFA were recorded, equivalent to 50-73% of total COD, in the thermophilic MBR effluent. The observed yield in the thermophilic MBR was 40% of that observed in the conventional MBR. PMID- 16221483 TI - Removal and recovery of Cr(VI) from polluted ground waters: a comparative study of ion-exchange technologies. AB - The focus of this work has been the study of Cr(VI) removal from ground waters and the simultaneous concentration for its reuse using three different technological alternatives: anion-exchange resins, liquid-liquid extraction assisted by hollow fibre membranes and emulsion pertraction. The viability of the considered objectives, i.e., Cr(VI) separation (<0.5 g/m3) and concentration for reuse (>20,000 g/m3) has been checked and a comparative analysis of the three technologies has been performed. Although the flexibility and ease of operation of non-dispersive solvent extraction, anion-exchange resins and emulsion pertraction lead to higher velocities of chromium removal, yet still maintaining similar concentration efficiencies. PMID- 16221484 TI - Norovirus pathway in water environment estimated by genetic analysis of strains from patients of gastroenteritis, sewage, treated wastewater, river water and oysters. AB - In this study, Norovirus (NV) capsid gene was detected from patients of gastroenteritis, domestic sewage, treated wastewater, river water and cultivated oysters in geographically close areas where all of samples were collected. In order to improve recovery efficiency of NVs from oysters, a new method using a spallation apparatus was developed. As a result, 18 of 30 oysters (60%) were positive for NV gene, while 7 of 30 (23%) oysters from the same sampling point were positive with the conventional ultracentrifugal method between November 2003 and February 2004. These results indicate that our new method exhibits the higher efficiency of recovering NVs than the conventional ultracentrifugal method. Six of 8 samples (75%) of river water were positive for NV gene between November 2003 and February 2004. Furthermore, 8 of 9 samples (89%) of treated wastewater and all 9 samples of sewage were positive for NV gene in the same period. These results indicated that treated wastewater would be one of the main sources for NV pollution in this area. The phylogenetic analysis in isolated NV capsid genes was conducted, in which high identities of gene sequences between NVs from patients, domestic sewage, river water and cultivated oysters were observed. These results implied that there would be a geographically associated circulation of NVs between human and cultivated oysters via water environment. It would be important to quantitatively analyze the moving pathway of NVs, which directly link to the development of a new scheme for preventing water environment and cultivated oysters from NV contamination. PMID- 16221485 TI - Mechanisms of gold recovery from aqueous solutions using a novel tannin gel adsorbent synthesized from natural condensed tannin. AB - We report a novel recovery system for gold (Au), which is one of the precious metals contained in electronic scrap, utilizing tannin gel particles. Tannin gel particles were prepared by a process of cross-linking of condensed tannin (wattle tannin), which is a ubiquitous and inexpensive natural material having many hydroxyl groups. The adsorption mechanism of gold onto tannin gel particles was elucidated: the adsorption of gold takes place through the reduction of trivalent gold ions to metallic gold on the surface of tannin gel particles, which is accompanied by the simultaneous oxidization of the hydroxyl groups of tannin gel. Additionally, the adsorption capacity of gold was found to be extremely high, 8000 mg-Au/g-dry gel. The outstanding characteristics of tannin gel particles for gold offers the possibility of efficient recovery of other precious metals. PMID- 16221486 TI - Role of nitric oxide in ovarian follicular development and egg production in Japanese quail (Coturnix coturnix japonica). AB - Role of nitric oxide (NO) in regulating the reproductive functions at hypothalamo hypophysealovarian axis in Japanese quail was studied. In first experiment, metabolites of NO, i.e. nitrite and nitrate (NO2 and NO3) were estimated together in hypothalamus, serum and ovarian follicles of good and poor layers. In the second experiment, different NO modulators such as L-arginine (L-Arg), sodium nitroprusside (SNP) and N(G)-nitro-L-arginine methyl ester, HCl (L-NAME) were administered to the birds. In the first experiment, significantly higher (P < 0.01) NO2 and NO3 levels in serum, hypothalamus and largest (F1) ovarian follicles were observed in good layers as compared to poor layers. Higher (P < 0.05) NO2 and NO3 concentration was observed in F1 follicles than smaller follicles (F2) only in good layers. The NO2 and NO3 concentration was significantly reduced (P < 0.05) in post ovulatory follicles (POFs) in comparison to F1 and F2 follicles. In the second experiment, the serum NO2 and NO3 concentrations were higher (P < 0.05) in the SNP, lower (P < 0.05) in the L-Name group and unchanged in the L-Arg treated group in comparison to control group. compared to control, L-Arg and SNP increased (P < 0.05) the hypothalamic NO2 and NO3 concentration where as L-NAME reduced (P < 0.05) these levels. The NO2 and NO3 concentration was increased (P < 0.05) as the follicle size increased and it was significantly reduced (P < 0.05) in POFs. The higher (P < 0.05) follicular NO2 and NO3 concentration was observed in L-Arg group in comparison to control group. Egg production was also found to be higher (P < 0.05) in L-Arg group whereas it was not different (P > 0.05) in SNP and L-NAME treated groups. The yolk weight and yolk to albumin ratio was reduced (P < 0.05) in L-NAME group in comparison to control group. It may be concluded from the present study that NO plays a key role in regulating follicular development, ovulatory mechanisms and egg production in Japanese quail. PMID- 16221487 TI - Effect of repeated administration of PGF2alpha in the early post partum period on the prevalence of clinical endometritis and probability of pregnancy at first insemination in lactating dairy cows. AB - Two experiments were performed to determine the effects of repeated administration of PGF2alpha in the immediate post partum period on the prevalence of clinical endometritis at 22 and 58 days post partum, and the probability of pregnancy at first insemination, in post partum dairy cows. In Experiment 1, 228 cows on day 7 post partum were used. Cows in Group 1 (n = 114) were treated twice with PGF2alpha (25 mg, im) 8 h apart on days 7 and 14 post partum, and only once on days 22 and 35 post partum. Cows in Group 2 (n = 114) served as untreated controls. Vaginoscopy and transrectal palpation of the genital tract were done on days 22 and 58 post partum. Cows in both groups were inseminated at estrus after a voluntary waiting period of 100 days. Pregnancy was determined by transrectal palpation between 45 and 50 days after insemination. Repeated administration of PGF2alpha in the immediate post partum period did not reduce the prevalence of clinical endometritis on days 22 or 58 post partum (65 versus 62% on day 22 and 28.3 versus 35.2% on day 58 in Groups 1 and 2, respectively). There was no significant difference in the probability of pregnancy at first insemination between cows in both groups. In Experiment 2, 418 cows on day 7 post partum were used. Cows in Group 1 (n = 209) were treated twice with PGF2alpha (25 mg, im) 8h apart on days 7 and 14 post partum, and only once on days 22 and 35 post partum. Cows in Group 2 (n = 209) served as untreated controls. Cows in both groups were subjected to the Presynch and Ovsynch protocols on days 49 and 75 post partum, respectively. Pregnancy was determined by transrectal ultrasonography between days 29 and 32 after timed insemination. There was no significant difference in the probability of pregnancy at first insemination between cows in both groups. In conclusion, repeated administration of PGF2alpha to cows in this herd had no effect on the prevalence of clinical endometritis at 22 and 58 days post partum, and that there was no effect on the probability of pregnancy after insemination at estrus among cows with a voluntary waiting period of > 100 days, or at timed AI at 85 days in milk when Presynch was employed. PMID- 16221488 TI - Effect of surface pre-treatment on durability of resin-based cements bonded to titanium. AB - OBJECTIVES: Titanium has in recent years become a material of great opportunities in dentistry and orthopedics. In this study, we searched for a method to effectively pre-treat titanium as part of an adhesive luting and/or resin veneering procedure. METHODS: Ti plates were treated with two different solutions of hydrochloric acid (0.1, 1N), 37wt% phosphoric acid or kept untreated (control). The tensile bond strength of the composite cement (Panavia Fluoro Cement, Kuraray) to the untreated and pre-treated Ti plates was determined without and after 20,000 thermo-cycles. XPS was used to chemically analyze the effect of the three Ti pre-treatments, as well as the interaction of 10 methacryloxydecyl dihydrogen phosphate (10-MDP; functional monomer of Panavia Fluoro Cement) with Ti treated with 1N HCl. RESULTS: Although no significant difference in immediate tensile bond strength was measured, thermo-cycling significantly decreased the bond strength of all experimental groups except for Ti treated with 1N HCl. No difference in surface roughness was found between untreated and treated Ti. XPS demonstrated that HCl effectively decontaminated Ti in a dose-dependent manner, while H(3)PO(4) was strongly adsorbed on the Ti surface. While the latter potentially inhibits adsorption of the phosphoric groups of the functional monomer 10-MDP, Ti pre-treated with 1N HCl improved the adsorption of 10-MDP as compared to untreated Ti. SIGNIFICANCE: It is concluded that 1N HCl effectively pre-treats Ti in order to make it more receptive to resin, while H(3)PO(4) should better not be used. PMID- 16221489 TI - Characterization of Portland cement for use as a dental restorative material. AB - OBJECTIVES: The aim of this study was to evaluate the suitability of fast-setting cement formulations based on Portland cement as dental core build-up materials using two different methods of testing compressive strength and evaluation of setting times. METHODS: Four fast-setting cements based on Portland cement and their four respective densified with small particle (DSP) mortars were tested for setting time, constitution of cement by EDAX, and compressive strength using International and British Standards. Ordinary Portland cement (OPC) was used as a control. RESULTS: All the fast-setting cements had a similar elemental composition to OPC and the setting times were less than 7 min. The compressive strength of OPC was different between the two methods (P<0.001). All the fast setting cements tested showed no difference in compressive strength regardless of the method of testing at 1 and 7 days (P>0.05), but the cylinders showed a lower compressive strength at 28 days (P<0.05). The OPC DSP mortar showed poorer compressive strength than OPC (P<0.01) at all times for cube testing but not for cylinder testing, where no difference was observed. The fast-setting DSP mortars had a lower compressive strength at 1 day (P<0.005) with both methods. At later times, there was no difference between the cements and DSP mortars for the cubes. SIGNIFICANCE: The pure fast-setting cements set in <7 min and were not susceptible to changes in the compressive strength testing procedure at 1 and 7 days but at 28 days all the fast-setting cements had a significantly higher strength with the test using cubes (P<0.05). A reduction in strength was observed at 28 days in cylinder testing. Most of the cements tested did not show encouraging strengths, however, one of the prototype cements tested could be a prospective dental restorative material. PMID- 16221490 TI - Elastic moduli of cast Ti-Au, Ti-Ag, and Ti-Cu alloys. AB - OBJECTIVES: This study investigated the effect of alloying titanium with gold, silver, or copper on the elastic properties of the alloys. METHODS: A series of binary titanium alloys was made with four concentrations of gold, silver, or copper (5, 10, 20, and 30 mass%) in an argon-arc melting furnace. The Young's moduli and Poisson's ratios of the alloy castings were determined with an ultrasonic-pulse method. The density of each alloy was previously measured by the Archimedes' principle. Results were analyzed using one-way ANOVA and the Scheffe's test. RESULTS: The densities of Ti-Au, Ti-Ag, and Ti-Cu alloys monotonically increased as the concentration of alloying elements increased. As the concentration of gold or silver increased to 20%, the Young's modulus significantly decreased, followed by a subsequent increase in value. As the concentration of copper increased, the Young's modulus monotonically increased. The Young's moduli of all the Ti-Cu alloys were significantly higher than that of the titanium. SIGNIFICANCE: The density of all the experimental alloys was virtually independent of the alloy phases, while the Young's moduli and Poisson's ratios of the alloys were dependent. The addition of gold or silver slightly reduced the Young's modulus of the titanium when the alloy phase was single alpha. The increase in the Young's modulus of the Ti-Cu alloys is probably due to the precipitation of intermetallic compound Ti2Cu. Copper turned out to be a moderate stiffener that gains a Young's modulus of titanium up to 20% at the copper concentration of 30 mass%. PMID- 16221491 TI - Low viscosity hydrogel of guar gum: preparation and physicochemical characterization. AB - Guar gum was cross-linked with glutaraldehyde and characterized by GPC, rheology, WADX, SEM and TGA. This guar gum is a galactomannan polysaccharide, that contains small amount of arabinose, glucose and uronic acid, besides galactose and mannose. The polymer has high molar mass, with Mw, Mn and Mv values of 2.0x10(6), 1.2x10(6) and 1.9x10(6)g/mol, respectively. The reticulation follows a slow process and lead to a viscosity increase of 40 times compared with the original gum solution. The final viscosity was similar to that of Hylan G-F 20, a hyaluronate derivative, commercially used in viscosupplementation treatment. The gel contains 95.6% of water and the amount of residual glutaraldehyde is much lower than the LD-50. Porous structure was detected by SEM and thermal stability was improved by the cross-linking. The low viscosity, the small amount of remained glutaraldehyde, and the thermal stability indicates that the guar hydrogel has potential to be applied as biomaterial with specific rheological requirements. PMID- 16221492 TI - Dextran-spermine-based polyplexes--evaluation of transgene expression and of local and systemic toxicity in mice. AB - Gene delivery using self-assembled polyplexes, formed between negatively charged nucleic acids and cationic polymers, have several drawbacks including low transgene expression and toxicity effects related to their positive charge. Recently, a novel cationic polymer based on dextran-spermine (D-SPM) was developed for gene delivery. This polymer showed systemic biodistribution upon local administration (intramuscular (i.m.) and intranasal (i.n.)) followed by transgene expression in organs remote from the site of injection (liver and lungs). Polyplexes based on D-SPM were administered both i.m. and i.n. to BALB/c female mice. LacZ expression in the liver, lungs, and muscles was assessed using whole-mount 5-bromo-4-chloro-3-indolyl beta-d-galactopyranoside (X-gal) staining and paraffin sectioning. The local toxicity in these organs was evaluated from hematoxylin and eosin stained sections. The systemic toxicity of the polymer and polyplexes was estimated by comparing the mice total weight, major organ weights, blood counts (primarily white blood cells (WBC) and platelets), and serum transaminases, to those of control animals (which received PBS). Transgene expression using D-SPM polyplexes was dependent upon the dosage and the polyplexes (+/-) charge ratio. Using the i.m. and i.n. routes of administration the transfection occurred primarily in the bronchial epithelial cells, pneumocytes, and bronchial alveoli of the lungs; in the muscle's fibrocytes; and in the liver's hepatocytes. Histopathological assays revealed mild toxicity in muscle and no abnormal findings in liver and lung. No systemic toxicity was obtained, as we did not find decrease in WBC count or platelet and no increase in serum transaminases. In addition, mice body weights and major organ weights were similar to the control group at both 2 and 28 days post-administration. This study demonstrates systemic transfection efficacy upon local administration of D SPM complexes with good tolerability and low toxicity. PMID- 16221493 TI - The in vivo and in vitro degradation behavior of poly(trimethylene carbonate). AB - The in vivo and in vitro degradation behavior of poly(trimethylene carbonate) (PTMC) polymers with number average molecular weights of 69 x 10(3), 89 x 10(3), 291 x 10(3) and 457 x 10(3)g/mol (respectively abbreviated as PTMC(69), PTMC(89), PTMC(291) and PTMC(457)) was investigated in detail. PTMC rods (3mm in diameter and 4mm in length) implanted in the femur and tibia of rabbits degraded by surface erosion. The mass loss of high molecular weight PTMC(457) specimens was 60wt% in 8 wks, whereas the mass loss of the lower molecular weight PTMC(89) specimens in the same period was 3 times lower. PTMC discs of different molecular weights immersed in lipase solutions (lipase from Thermomyces lanuginosus) degraded by surface erosion as well. The mass and thickness of high molecular weight PTMC(291) discs decreased linearly in time with an erosion rate of 6.7 microm/d. The erosion rate of the lower molecular weight PTMC(69) specimens was only 1.4mum/d. It is suggested that the more hydrophilic surface of the PTMC(69) specimens prevents the enzyme from acquiring a (hyper)active conformation. When PTMC discs were immersed in media varying in pH from 1 to 13, the non-enzymatic hydrolysis was extremely slow for both the high and low molecular weight samples. It can be concluded that enzymatic degradation plays an important role in the surface erosion of PTMC in vivo. PMID- 16221494 TI - PCL microparticle-dispersed PLGA solution as a potential injectable urethral bulking agent. AB - The PCL microparticle-dispersed PLGA solutions were prepared as a potential injectable urethral bulking agent. The mixture solutions were prepared by mixing polycarprolactone (PCL) microparticles (diameter, 100 approximately 200mum; fabricated by a temperature-induced phase transition method) and poly(dl-lactic co-glycolic acid) (PLGA) solution (dissolved in tetraglycol to 10wt%) with different PCL microparticle to PLGA solution ratio. The mixture solution was solidified by the precipitation of PLGA when the solution was contact with water. In contact with water, the PCL microparticles exhibited a well-packed structure entrapped in a solidified porous PLGA matrix, which can effectively prevent the microparticle migration in the body and retain its initial volume even after PLGA matrix degradation. The PCL microparticle-dispersed PLGA solution (particle to solution ratio, 45/55 (w/v)) was easily injected through 18G needle into back of hairless mouse (subcutaneously) and stably located at the apply site. The surrounding tissue including blood vessel were gradually infiltrated into the implant up to 8 weeks without the initial injected volume change and with little inflammatory response. The PCL microparticle-dispersed PLGA solution may be a good candidate as an injectable bulking agent for the treatment of urinary incontinence owing to its good injectability, volume retention potential as well as biocompatibility. PMID- 16221495 TI - Role of complement and B lymphocytes in Sjogren's syndrome-like autoimmune exocrinopathy of NOD.B10-H2b mice. AB - Sjogren's syndrome (SjS) is a human autoimmune disease characterized by the loss of exocrine function as a result of a chronic immune attack directed primarily against the salivary and lacrimal glands leading to xerostomia (dry mouth) and xerophthalmia (dry eyes). NOD.B10-H2b mice manifest many features of SjS, exhibiting exocrine gland dysfunction concomitant with leukocyte infiltration of the salivary and lacrimal glands. Recent studies have shown that both SjS patients and NOD.B10-H2b mice exhibit increased B lymphocyte survival, B cell hyper-reactivity and hyper-gammaglobulinemia with high production of autoantibodies. To study the possible influence of complement on the development and expression of SjS-like disease of the NOD.B10-H2b, we have utilized a prophylactic treatment with CVF known to interfere with the action of complement C3 factor. NOD.B10-H2b mice, injected with CVF starting at 10 weeks of age, a time when leukocyte infiltration is expected to begin, failed to develop salivary dysfunction out to 24 weeks of age, a time when reduced salivary flow rates are known to occur in non-treated animals. Concomitant with retention of salivary exocrine function, CVF-treated mice showed reduced levels of leukocytic infiltration, reduction of anti-nuclear autoantibodies and major alterations in the B lymphocyte profiles while maintaining general pathophysiological measures of disease. These data suggest that C3 plays a significant role in development and onset of SjS-like disease, yet additional studies need to be carried out to identify the precise mode of action. PMID- 16221496 TI - A reliable RT-PCR-ELISA method for the detection of infectious pancreatic necrosis virus (IPNV) in farmed rainbow trout. AB - A new method, termed RT-PCR-ELISA, was evaluated for ease of use, reliability and sensitivity when detecting infectious pancreatic necrosis virus (IPNV) present in trout kidney tissue. The method had comparable sensitivity to existing PCR assays and could successfully detect 1.5 x 10(4) pfu IPNV in artificially contaminated trout kidney samples. The technique was easily established in a new laboratory and required no specialised equipment. The method had a high sample throughput capable of screening 96 samples per run, making the technique extremely time efficient. The RT-PCR-ELISA is a safe, quick, reliable technique, which has the potential for use as a standard virus detection method. PMID- 16221498 TI - The neutral red lysosomal retention assay and Comet assay on haemolymph cells from mussels (Mytilus edulis) and fish (Symphodus melops) exposed to styrene. AB - Despite the extensive transport of chemicals at sea, there is current lack of knowledge of the fate and effects of many of them on the marine biota. The current regulation that follows the GESAMP-MARPOL classification is mainly based on ecotoxicity assessment from fresh water based studies. Repetitive spills in marine coastal environment from tanker ship loaded with several thousand tonnes of chemicals raised concern about whether the existing freshwater data location can be used to predict the behaviour and the environmental effects of contaminants in marine surroundings. There is a general lack of information of the fate of chemicals at sea. A deviating pattern in marine environment from that in freshwater may have significant consequences for the counteracting actions taken to fight the spill, on staff working on the site of spill as well as on marine life present in the vicinity of the accident. In the present article, an environmental effect study of styrene was conducted as part of the ECOPEL program. We report some biological effects of styrene in laboratory-exposed marine organisms. Styrene was continuously supplied at a nominal concentration of 2mg L(-1) over 7 days to both mussels (Mytilus edulis) and fish (Symphodus mellops). At the end of this period, DNA damage was assessed by the Comet assay performed on blood (fish) and haemolymph (mussel) cells. In mussels, the lysosomal membrane stability was additionally assessed by the neutral red retention time assay (NRRT). Significant biological responses were observed over the studied period in both organisms with these two tests. Hence, the results favour the use of a biomarker-based approach to assess the health conditions in case of spill. PMID- 16221497 TI - Heterozygous mice with Ric-8 mutation exhibit impaired spatial memory and decreased anxiety. AB - Ric-8 is a guanine nucleotide exchange factor for a subset of Galpha proteins and it is required to maintain Galpha(q) and the Galpha(s) pathways in functional state. In adult mice Ric-8 is expressed in regions involved in the regulation of behavior (neocortex, cingulate cortex and hippocampus). As Ric-8 is shown to regulate neuronal transmitter release, the aim of present study was to perform behavioral analysis of ric-8 mutant. Homozygous (-/-) ric-8 mutant mice are not viable and die in early embryonic development, therefore for behavioral analysis heterozygous (+/-) ric-8 mutant mice were used. We found decreased anxiety of ric 8 heterozygous mice in light-dark compartment test where mutant mice significantly avoided the light compartment. In spatial learning paradigm (Morris water maze) the performance of ric-8 (+/-) mice was impaired. Namely, in the reversal test, ric-8 (+/-) mice exhibited significant delay to find the hidden platform compared to wild-type (wt) littermates. We did not find differences in the behavioral tests reflecting the motor abilities of mice (motor activity, rota rod). Therefore, described alterations seem to be specific for anxiety and spatial learning. Based on these results we can conclude the importance of ric-8 in the regulation of memory and emotional behavior. PMID- 16221499 TI - Medial prefrontal cortex lesions inhibit reflex micturition in anethetized rats. AB - The medial prefrontal cortex is thought to participate in the control of micturition and urinary continence, based on evidence from clinical reports, but its exact role is not fully understood. This study investigated whether ibotenic acid lesions of the medial prefrontal cortex would influence volume-evoked micturition in urethane-anesthetized rats. The incidence and amplitude of bladder contractions were recorded during continuous saline infusion (0.1 ml/min) immediately before and 1 week after ibotenic acid (0.5 microg) or vehicle (0.5 microl) was injected into the medial prefrontal cortex. Vehicle injection did not change the incidence or amplitude of bladder contractions compared to pre injection values. Ibotenic acid lesions prolonged the time interval between bladder contractions significantly although it did not affect the amplitude of bladder contractions. Histological analysis revealed that ibotenic acid lesions were restricted primarily to the anterior cingulate and prelimbic cortices. Larger ibotenic acid lesions extending ventrally into the infralimbic cortex produced a variable response but did not change the overall incidence or amplitude of bladder contractions significantly. These data indicate that the medial prefrontal cortex influences the timing of bladder contractions but does not affect contraction amplitudes. PMID- 16221500 TI - Identification of StiR, the first regulator of secondary metabolite formation in the myxobacterium Cystobacter fuscus Cb f17.1. AB - Myxobacteria are well established as proficient producers of natural products with numerous biological activities. Although some knowledge has been gained regarding the biosynthesis of secondary metabolites in this class of bacteria, almost nothing is known about the underlying regulatory mechanisms. In order to identify regulatory elements, we submitted the argyrin and stigmatellin producer Cystobacter fuscus to a random transposon mutagenesis strategy and screened 1,000 mutants for the occurrence of strains showing remarkably increased or decreased production of these compounds. In addition to the identification of the stigmatellin biosynthetic gene cluster, a novel positive regulator (stiR) of stigmatellin production was identified after transposon recovery. In order to exclude secondary mutagenesis effects, a double cross-over mutagenesis strategy was applied to the strain resulting in the repeated generation of the transposon genotype. This strain was shown to be equally effected in natural product biosynthesis. PMID- 16221501 TI - Recovery of recombinant beta-glucosidase by expanded bed adsorption from Pichia pastoris high-cell-density culture broth. AB - Methanol limited fed-batch cultivation was applied for production of a plant derived beta-glucosidase by Pichia pastoris. The beta-glucosidase was recovered by expanded bed adsorption chromatography applied to the whole culture broth. The new Streamline Direct HST1 adsorbent was compared with Streamline SP. Higher bead density made it possible to operate at two times higher feedstock concentration and at two times higher flow velocity. The higher binding capacity in the conductivity range 0-48 mS cm(-1) of Streamline Direct HST1 might be caused by the more complex interaction of multi-modal ligand in Streamline Direct HST1 compared to the single sulphonyl group in Streamline SP. Harsher elution condition had to be applied for dissociation of beta-glucosidase from Streamline Direct HST1 due to stronger binding interaction. The 5% dynamic binding capacity was 160 times higher for Streamline Direct HST1 compared to Streamline SP. The yield of beta-glucosidase on Streamline Direct HST1 (74%) was significantly higher than on Streamline SP (48%). Furthermore, beta-glucosidase was purified with a factor of 4.1 and concentrated with a factor of 17 on Streamline Direct HST1 while corresponding parameters were half of these values for Streamline SP. Thus, for all investigated parameters Streamline Direct HST1 was a more suitable adsorbent for recovery of recombinant beta-glucosidase from unclarified P. pastoris high-cell-density cultivation broth. PMID- 16221502 TI - Osteopontin expression in normal and fibrotic liver. altered liver healing in osteopontin-deficient mice. AB - BACKGROUND/AIMS: Osteopontin has been implicated in numerous physiopathological events. Osteopontin expression in normal and fibrotic liver and liver fibrogenesis in osteopontin-deficient mice were studied. METHODS: Fibrosis was induced in mice and rats by carbon tetrachloride (CCl4) treatment or bile duct ligation. The liver was used for conventional histology, osteopontin immunohistochemistry and in situ hybridization, or protein and RNA extraction. In mice, necrotic areas and fibrosis were evaluated by quantitative image analysis. RESULTS: In normal liver, osteopontin mRNA expression was very low. After CCl4 treatment or bile duct ligation, osteopontin mRNA expression was increased. Osteopontin was expressed by biliary epithelial cells in normal and fibrotic liver. Soon after the beginning of the CCl4 treatment, osteopontin was also present in inflammatory cells of the necrotic areas. In osteopontin-deficient mice, necrotic areas after a single dose of CCl4, and fibrosis after chronic CCl4 treatment were significantly increased as compared with wild-type treated mice. CONCLUSIONS: Our results show that osteopontin expression increases during liver fibrogenesis. Furthermore, osteopontin-deficient mice were more susceptible to CCl4 treatment, displaying more necrosis during the initial steps (probably due to a deficiency in nitric oxide production) and more fibrosis thereafter. The increase in osteopontin expression observed during liver fibrogenesis may play a protective role. PMID- 16221503 TI - Regulatory effects of tumor necrosis factor-alpha and interleukin-6 on HAMP expression in iron loaded rat hepatocytes. AB - BACKGROUND/AIMS: To study the effect of iron and proinflammatory cytokines on the expression of HAMP and other iron regulatory genes in primary rat hepatocytes. METHODS: Primary hepatocytes from rats fed a control or iron-enriched diet were plated on extracellular matrix and incubated with inflammatory stimuli in the presence or absence of serum. Cells were also incubated with desferrioxamine or ferric ammonium citrate. mRNA levels were determined by Real-Time PCR. RESULTS: Hepatocytes from control rats increased their HAMP expression during culturing, whereas the opposite was seen in hepatocytes from carbonyl-iron loaded animals. In the presence of serum, tumor necrosis factor-alpha, lipopolysaccharide and interleukin-6 increased HAMP expression in hepatocytes from both control and iron loaded rats. Under serum-free conditions only tumor necrosis factor-alpha increased HAMP mRNA levels. Desferrioxamine and ferric ammonium citrate decreased HAMP gene expression. Tumor necrosis factor-alpha significantly increased mRNA levels of TfR2 and decreased those of DMT1 and IREG1. CONCLUSIONS: HAMP expression differs in cultured as compared with freshly isolated hepatocytes, and decreases in iron-loaded hepatocytes in serum free-media, suggesting that additional serum factors influence HAMP expression. Tumor necrosis factor-alpha regulates the mRNA levels of HAMP, IREG1, DMT1 and TfR2 in cultured hepatocytes from both iron-loaded and control animals. PMID- 16221504 TI - Prevalence and content of written ethics policies on euthanasia in Catholic healthcare institutions in Belgium (Flanders). AB - BACKGROUND: Euthanasia is performed worldwide, regardless of the existence of laws governing it. Belgium became the second country in the world to enact a law on euthanasia in 2002. Healthcare institutions bear responsibility for guaranteeing the quality of care for patients at the end of life, and for ensuring support for caregivers involved. Therefore, institutional ethics policies on end-of-life decision-making, especially on euthanasia, may be useful. METHODS: A cross-sectional mail survey of general directors of Catholic hospitals and nursing homes in Belgium was used to describe the prevalence and content of written ethics policies for competent terminally ill, incompetent terminally ill, and non-terminally ill patients. RESULTS: Of the 298 targeted institutions, 81% of hospitals and 62% of nursing homes returned complete questionnaires. Of these, 79% of hospitals and 30% of nursing homes had a written ethics policy on euthanasia. Of hospitals 83% and of nursing homes 85% permitted euthanasia for competent terminally ill patients only in exceptional cases in accordance with legal due care criteria and provisions outlined by the palliative filter procedure. Euthanasia for incompetent terminally ill patients was prohibited by 27% of the hospitals and by 60% of the nursing homes. For non-terminally ill patients, these figures were 43 and 64%, respectively. CONCLUSIONS: Catholic healthcare institutions in Belgium (Flanders) made great efforts to develop written ethics policies on euthanasia. Only a small group of institutions completely prohibited euthanasia. Most of the institutions considered euthanasia to be an option if all possible alternatives (e.g., palliative filter procedure, which contains more rigorous criteria than those in the Belgian Euthanasia Act), have been thoroughly investigated. PMID- 16221505 TI - Tau is central in the genetic Alzheimer-frontotemporal dementia spectrum. AB - In contrast to the common and genetically complex senile form of Alzheimer's disease (AD), the molecular genetic dissection of inherited presenile dementias has given important mechanistic insights into the pathogenesis of degenerative brain disease. Here, we focus on recent genotype-phenotype correlative studies in presenile AD and the frontotemporal dementia (FTD) complex of disorders. Together, these studies suggest that AD and FTD are linked in a genetic spectrum of presenile degenerative brain disorders in which tau appears to be the central player. PMID- 16221506 TI - A case-mix model for monitoring of postoperative mortality after surgery for lung cancer. AB - BACKGROUND: Postoperative mortality (POM) after surgery for lung cancer has been proposed as a performance indicator. Information on the size of the risk and its prognostic factors is needed to serve as a reference standard. PATIENTS AND METHODS: Electronic records from the Rotterdam Cancer Registry (n=2337) and the Thames Cancer Registry (n=3772) were retrieved and analysed by sex, age, period, histology, region and extent of surgery. Multivariable logistic regression analysis was used to determine prognostic factors, to calculate odds ratios (OR) and to develop a case-mix model. RESULTS: POM was 4.2% (n=257) on average and increased with age from 1.7% for patients younger than 60 years up to 9.4% for octogenarians. After lobectomy, POM was 2.9% against 6.0% and 9.5% after pneumonectomy left and right, respectively. Multivariable analysis showed higher risk for men (OR=1.4) and lower risk for adenocarcinoma (OR=0.6). CONCLUSIONS: The final prediction model supports comparison and monitoring of POM rates for lung cancer. Only a limited number of risk factors need to be registered to allow adjustment for case-mix. PMID- 16221507 TI - Environmental regulation of biofilm development in methicillin-resistant and methicillin-susceptible Staphylococcus aureus clinical isolates. PMID- 16221508 TI - Impact of deep infection after hip fracture surgery on function and mortality. AB - Our aim was to evaluate the impact of deep wound infection after hip fracture surgery on functional outcome and mortality. Two thousand, two hundred and seventy-six consecutive surgically treated patients with non-pathological fractures, aged over 50 years (mean age 78.3 years), were followed up prospectively. The 29 patients who had deep infection (rate 1.3%) and who received revision surgery were compared with controls without infection, matched for age, sex, residential status at fracture, fracture type, treatment method and walking ability. When the functionality of hip fracture patients was evaluated four months after the primary operation, the patients with deep wound infection had impaired walking ability (P=0.039) and required walking aids, including wheelchairs, more often than the control patients (48% vs 20.8%, P=0.022). At four months, the mean duration of hospitalization at the primary hospital was significantly longer for the cases than the controls (P<0.001). Diabetes was more common in the patients with deep infection (P=0.038). Staphylococcus aureus was the most commonly isolated micro-organism, and it was associated with higher one year mortality (57%) than the other bacteria (14.3%, P=0.014); the overall mortality of the infected patients and controls was 34.5% and 24.1% at one year, respectively (P=0.508). In conclusion, deep infection after a hip fracture operation impairs short-term functional outcome and slightly increases mortality, with an excess mortality rate of 10.4%. PMID- 16221509 TI - Evaluation of a 2% chlorhexidine gluconate in 70% isopropyl alcohol skin disinfectant. AB - The efficacy of a new skin disinfectant, 2% (w/v) chlorhexidine gluconate (CHG) in 70% (v/v) isopropyl alcohol (IPA) (ChloraPrep), was compared with five commonly used skin disinfectants against Staphylococcus epidermidis RP62A in the presence or absence of protein, utilizing quantitative time-kill suspension and carrier tests. All six disinfectants [70% (v/v) IPA, 0.5% (w/v) aqueous CHG, 2% (w/v) aqueous CHG, 0.5% (w/v) CHG in 70% (v/v) IPA and 10% (w/v) aqueous povidone iodine (PI)] achieved a log(10) reduction factor of 5, in colony-forming units/mL, in a suspension test (exposure time 30s) in the presence and absence of 10% human serum. Subsequent challenges of S. epidermidis RP62A in a biofilm (with and without human serum) demonstrated reduced bactericidal activity. Overall, the most effective skin disinfectants tested against S. epidermidis RP62A were 2% (w/v) CHG in 70% IPA and 10% (w/v) PI. These results suggest that enhanced skin antisepsis may be achieved with 2% (w/v) CHG in 70% (v/v) IPA compared with the three commonly used CHG preparations [0.5% (w/v) aqueous CHG, 2% (w/v) aqueous CHG and 0.5% (w/v) CHG in 70% (v/v) IPA]. PMID- 16221510 TI - Nosocomial infections in a Dutch neonatal intensive care unit: surveillance study with definitions for infection specifically adapted for neonates. AB - The incidence of nosocomial infection in neonatal intensive care units (NICUs) is high compared with other wards. However, no definitions for hospital-acquired infection are available for NICUs. The aim of this study was to measure the incidence of such infections and to identify risk factors in the NICU of the VU University Medical Center, which serves as a level III regional NICU. For this purpose, a prospective surveillance was performed in 1998-2000. We designed definitions by adjusting the current definitions of the Centers for Disease Control and Prevention (CDC) for children <1 year of age. Birth weight was stratified into four categories and other baseline risk factors were dichotomized. Analysis of risk factors was performed by Cox regression with time dependent variables. The relationship between the Clinical Risk Index for Babies (CRIB) and nosocomial infection was investigated. Furthermore, for a random sample of cases, we determined whether bloodstream infection and pneumonia would also have been identified with the CDC definitions. Seven hundred and forty-two neonates were included in the study. One hundred and ninety-one neonates developed 264 infections. Bloodstream infection (N=138, 14.9/1000 patient-days) and pneumonia (N=69, 7.5/1000 patient-days) were the most common infections. Of bloodstream infections, 59% were caused by coagulase-negative staphylococci; in 21% of neonates, blood cultures remained negative. In 25% of pneumonias, Enterobacteriaceae were the causative micro-organisms; 26% of cultures remained negative. Compared with the Nosocomial Infections Surveillance System (NNIS) of the CDC, our device utilization ratios and device-associated nosocomial infection rates were high. The main risk factors for bloodstream infection were birth weight [hazard ratio (HR) 1.79, 95% confidence interval (CI) 1.45-2.17] and parenteral feeding with hospital-pharmacy-produced, all-in-one mixture 'Minimix' (HR 3.69, 95%CI 2.03-6.69); administration of intravenous antibiotics (HR 0.39, 95%CI 0.26-0.56) was a protective risk factor. The main risk factors for pneumonia were low birth weight (HR 1.37, 95%CI 1.01-1.85) and mechanical ventilation (HR 9.69, 95%CI 4.60-20.4); intravenous antibiotics were protective (HR 0.37, 95%CI 0.21-0.64). In a subcohort of 232 very-low-birthweight neonates, the CRIB was not predictive for infection. With the CDC criteria, only 75% (21/28) of bloodstream infections and 87.5% of pneumonias (21/24) would have been identified. In conclusion, our local nosocomial infection rates are high compared with those of NICUs participating in the NNIS. This can be partially explained by: (1) the use of our definitions for nosocomial infection, which are more suitable for this patient category; and (2) the high device utilization ratios. PMID- 16221511 TI - Cost comparison of rabies pre-exposure vaccination with post-exposure treatment in Thai children. AB - Thailand is a canine rabies endemic country with an annual prevalence above 1,000 reported animals diagnosed rabid . Over 345,000 humans are treated for possible rabies exposures annually . Lack of perception of the disease burden, social, cultural and traditional beliefs play an important role in the failure of canine rabies control. It is unfortunate that health care budgets are increasingly allocated to human post-exposure treatment rather than to the eradication of rabies in the canine animal vector. Children under the age of 15 years represent up to one-half of dog bite victims and of human rabies deaths, but accurate data of dog bite prevalence are not available . Large scale pre-exposure immunization of children has been advocated but financial and logistic barriers have hindered implementation. This study analyzes direct medical costs of pre-exposure vaccination (PREP) as a human rabies preventive strategy, against the cost of post-exposure prophylaxis (PEP) in Thai children. Three pre- and post-exposure vaccine regimens are in use and this impacts on cost calculations. It was found that costs of both strategies, PREP of children or PEP of exposed, become equal when the dog bite incidence is 2-30%; depending on which post-exposure treatment regimens (PEP) are used. PMID- 16221512 TI - A comparative immunopathological study of injection site reactions in salmonids following intraperitoneal injection with oil-adjuvanted vaccines. AB - An adequate understanding of the inflammatory mechanisms following intraperitoneally-injected oil-adjuvanted vaccines in fish is required for the improvement of the quality of vaccines. We report the findings of a study in which the importance of neutrophils, macrophages, lymphocytes and mast cells (MCs) in early inflammatory reactions of Atlantic salmon were examined in comparison with those of rainbow trout and grayling. The inflamed areas in the pylorus/pancreas area (injection site) were sequentially measured using computer assisted microscopy, parallel with the quantification of leucocytes from 2 to 16 weeks post-vaccination. Antigen retention was also evaluated. Results show that inflammation in Atlantic salmon was slow to develop and was associated with a prolonged initial dominance of neutrophils. In contrast, the onset in rainbow trout was rapid and strong with early macrophage domination. The inflammatory and cellular response in grayling was initially in-between but progressed to be the strongest at 16 weeks. The results suggest that slow development of inflammation in Atlantic salmon and grayling leads to delayed sequestration of antigens resulting in chronically active inflammation that attempts to remove the antigens from the injection site. The strong onset in rainbow trout leads to rapid breakdown and removal of antigens resulting in early resolution. PMID- 16221513 TI - Immunization with gp120-depleted whole killed HIV immunogen and a second generation CpG DNA elicits strong HIV-specific responses in mice. AB - HIV-1 Immunogen is a gp120-depleted whole killed virus vaccine candidate formulated with Incomplete Freund's Adjuvant (HIV-IFA). We evaluated in a mouse model the immunogenicity of HIV-IFA by itself and when combined with HYB2055, an immunomodulatory oligonucleotide consisting of a novel DNA structure and synthetic CpR immunostimulatory motif, as an adjuvant. C57/BL6 mice were immunized with HIV-IFA alone or combined with HYB2055. Mice treated with HYB2055 or with PBS were used as controls. Compared to HIV-IFA alone, immunization with HIV-IFA and HYB2055 combination elicited strong production of HIV- and p24 specific IFNgamma, RANTES, MIP 1alpha, and MIP 1beta, as well as high titers of HIV- and p24-specific antibodies. Inclusion of HYB2055 also reduced levels of IL 5 produced by HIV-IFA alone. HYB2055 enhances the immunogenicity of HIV-IFA and shifts responses towards a type 1 cytokine profile. The immune enhancing effects of HYB2055 adjuvant were dose-dependent. These findings warrant clinical evaluation of the HIV-1 immunogen/HYB2055 candidate as a therapeutic vaccine for HIV-1 infected patients. PMID- 16221514 TI - Inter-carpal soft tissue entrapment. A possible explanation for chronic dorsal wrist pain. AB - This retrospective study evaluates the surgical treatment of a group of patients with unknown chronic dorsal wrist pain. The cause of their symptoms was interpreted as a painful entrapment of fibrous tissue in the radio-carpal and inter-carpal joints during specific movements. Between 1997 and 2001, 30 patients were treated by surgical excision of this tissue and 26 patients were traced for follow-up. Twenty-three patients were symptom free, or experienced major benefit from surgery. Wrist function measurements using the VAS scale showed improvement in 24 patients. Microscopic examination of the removed specimen shows fibrous tissue with non-specific changes. Inter-carpal soft tissue entrapment can explain the typical clinical findings in some patients with unknown chronic dorsal wrist pain. After careful selection, surgical excision of all entrapped tissue in the radio-carpal and mid-carpal joint may give relief of pain and improvement of wrist function. PMID- 16221515 TI - A comparative appraisal of the relationship of education, income and housing tenure with less than good health among the elderly in Europe. AB - The objective of this study was to determine the strength of various socio economic indicators for predicting less than good health among elderly people aged 60-79 years. Data were obtained from national health surveys from 10 European countries. Education, income and housing tenure were examined in relation to less than good health using standardised prevalence rates and (multiple) logistic regression analyses. The results illustrated that there are substantial health differences among the elderly according to education and income in each country. Both education and income (with men) showed a strong independent relationship with health status. Health differences according to housing tenure were generally somewhat smaller. However, in Great Britain and the Netherlands housing tenure demonstrated large health differences, even after adjustment for education and income. It is recommended that more refined socio economic measures are developed and that in the meantime both education and income are used when studying socio-economic health differences among the elderly. In some countries, like Great Britain and the Netherlands, however, housing tenure has an additional value. PMID- 16221516 TI - Prooxidative effect of copper--metallothionein in the acute cytotoxicity of hydrogen peroxide in Ehrlich ascites tumour cells. AB - This study was concerned with the role of copper (Cu) and Cu-metallothionein (Cu MT) in oxidative stress. Hydrogen peroxide (H(2)O(2))-induced oxidative injury was examined in Ehrlich ascites tumour cells isolated from host mice pretreated with 0, 1 or 2mg of CuSO(4) (ip) 24h earlier. Control Ehrlich cells contained low levels of Cu and Cu treatment produced dose-related increases in cellular Cu and Cu-MT levels and corresponding increases in sensitivity to oxidative toxicity of H(2)O(2) (LC(50), cell blebbing, lipid peroxidation, GSH depletion, and increase in intracellular free [Ca(2+)](i)). Hydrogen peroxide treatment also resulted in the oxidation of MT thiolates, reduction in the binding of Cu to MT resulting in translocation of Cu to other subcellular sites. d-penicillamine, a Cu-chelating agent, obliterated the sensitization effect of Cu-pretreatment and reduced the redistribution of MT-bound Cu, suggesting the participation of Cu ions derived from MT in promoting oxidant stress. Additional experiments with desferoxamine and mannitol have revealed the involvement of a Cu-dependent Fenton reaction in the mediation of the prooxidative effect of Cu-MT. These data suggest that cells with high levels of Cu-MT may be particularly susceptible to oxidative stress. PMID- 16221517 TI - Outcome of direct restorations placed within the general dental services in England and Wales (Part 2): variation by patients' characteristics. AB - AIM: It is the purpose of this paper to investigate the outcome of direct placement restorations provided within the General Dental Services in England and Wales, and to identify the patient factors which may affect this using a database derived from patient treatment data at the Dental Practice Board. METHODS: For this work, survival of a restoration was considered to be the time between the date of completion of the course of treatment in which it was placed and the date of acceptance of the course of treatment when the next tooth-specific treatment was carried out on the same tooth. A modified version of Kaplan-Meier statistical methodology was used to plot survival curves for restorations in different subgroups of patients within the population of patients for whom data were available. RESULTS: The results indicated that patients' gender was of little significance in the long-term survival of restorations, but patient age had a significant effect, with the restorations of older patients surviving less well than those of younger patients (P<0.001). The charge-paying status of the patient had a statistically significant but small effect (P<0.001), with survival for charge-payers being slightly higher throughout the observation period. There was a strong relationship between attendance frequency and survival time, with restorations of more frequent attenders performing less well than those of less frequent attenders (P<0.001). CONCLUSIONS: Patient factors such as age, charge paying status, interval between courses of treatment, continuity of care by one dentist, and annual gross fees incurred, have all been found to influence the survival of directly placed restorations. PMID- 16221518 TI - Outcome of direct restorations placed within the general dental services in England and Wales (Part 4): influence of time and place. AB - AIM: It is aim of this paper to investigate the outcome of direct-placement restorations provided within the General Dental Services in England and Wales, in relation to the factors related to time and place of restoration placement. METHODS: A modified version of Kaplan-Meier statistical methodology was used to plot survival curves for restorations placed at different times and places, within the population of patients for whom data were available. RESULTS: The results indicated that overall, restoration survival time to re-intervention has declined over the last decade, but that most of the decline can be accounted for by the change in the relative numbers of amalgam and other direct restorations. Restoration survival to re-intervention is independent of whether the water is fluoridated in the area of the dentist's surgery address. Variation with region is not great, but restorations placed in Wales survived to re-intervention significantly longer than those in England (P<0.0001). CONCLUSIONS: Overall, there was some evidence of a reduction in survival times with time. Restorations placed in Wales survived longer to re-intervention than restorations placed in England. Restoration survival was not dependent on whether the dentist who placed the restoration practised in an area in which the water was fluoridated. Performance of amalgam restorations was not found to have altered significantly with time, but performance of composite and glass ionomer deteriorated with time. PMID- 16221519 TI - Outcome of direct restorations placed within the general dental services in England and Wales (Part 1): variation by type of restoration and re-intervention. AB - AIM: It is the aim of this paper to consider the dental factors associated with the need for re-intervention on a restoration, such as the tooth position, size of cavity, and restoration material. METHODS: Patients whose data were included in this study were those whose birthdays were included within a set of randomly selected dates, one of which was chosen in each possible year of birth. The restoration records consisted of all those records containing directly placed restorations which related to courses of treatment of patients 18 years or older with last date on the claim form after 31st December 1990, and with date of acceptance after September 1990 and before January 2002. For each tooth treated with a direct restoration the subsequent history of intervention on that tooth was consulted, and the next date of intervention, if any could be found in the extended data set, was obtained. Thus a data set was created of direct restorations with their dates of placement and their dates, if any, of re intervention. RESULTS: Data for over 80,000 different adult patients were analyzed, of whom 46% were male and 54% female. A total of 503,965 tooth restoration occasions were obtained from the data over a period of eleven years. Single surface amalgam restorations were found to have the longest survival --58% at 10 years, and glass ionomer the shortest 38% at 10 years. Factors which were found to reduce restoration outcome included involvement of the incisal angle in composite restorations--this resulted in a reduction in median survival of around two years--and the placement of pins in a restoration. The presence of a root filling was also found to reduce the survival of restorations in the crown of the root filled tooth. CONCLUSIONS: Small amalgam restorations have longer survival times before re-intervention than large amalgam restorations such as MOD. Composite and glass ionomer restorations perform less well than amalgam restorations. Pin placement and root filling reduce the survival time of restorations. PMID- 16221520 TI - The International Liaison Committee on Resuscitation (ILCOR)-past and present: compiled by the Founding Members of the International Liaison Committee on Resuscitation. PMID- 16221521 TI - Cardiac arrest and resuscitation with an automatic mechanical chest compression device (LUCAS) due to anaphylaxis of a woman receiving caesarean section because of pre-eclampsia. AB - We report a case of anaphylaxis with pulseless electrical activity (PEA)(verified by ECG and a radial intra-arterial line) in a 30-year-old woman who received 3G Promiten (dextran-1) and a prophylactic intra-venous infusion of Macrodex (dextran) for postoperative thromboembolism during caesarean section for pre eclampsia in the 24th week of gestation. Manual chest compressions, followed by mechanical chest compressions (LUCAS, Jolife, Lund, Sweden), were performed for 50min before restoration of spontaneous circulation (ROSC). She awoke the next day with no sequelae. She had some suction cup marks on the sternum but otherwise no complications of the chest compressions. At follow up by phone 1 month later, she and her baby were doing well. PMID- 16221522 TI - Inconsistencies in cardiac arrest reporting. AB - Data relating to survival from in-hospital cardiac arrest are used to audit staff performance and to help to determine whether new resuscitation techniques are effective. Individual studies into outcome from cardiac arrest have defined inclusion and exclusion criteria, but no such national criteria have been published to enable constant auditing of cardiac arrests. The aim of this survey was to investigate the consistency with which in-hospital cardiac arrests are recorded throughout the United Kingdom. Such data are, almost universally, collected by Resuscitation Officers (RO). A questionnaire was sent to ROs across the UK asking them to state how they would interpret and categorise hypothetical, but nonetheless typical, clinical situations involving a cardiac arrest team being called. These included an event where the patient had regained consciousness prior to the arrival of the cardiac team and also an event where rigor mortis was already present and the resuscitation promptly abandoned upon the arrival of the cardiac arrest team. The percentage survival to discharge of adult cardiac arrests for each hospital was also requested. This identified whether inclusion or exclusion of certain clinical events may have influenced cardiac arrest survival figures for that hospital. It is clear from this study that in-hospital clinical events when a cardiac arrest team is called are audited with a great deal of inconsistency. Some events, such as a patient who has rigor mortis, are excluded as a false or inappropriate call in some hospitals and included as an unsuccessful resuscitation in others. There is a need for guidance on the inclusion and exclusion criteria for auditing of cardiac arrests so that meaningful data can be obtained from across the UK and useful conclusions drawn. The situation at present will result in data being audited that are of limited use. In the era of evidence-based medicine, it seems vital to obtain accurate cardiac arrest survival figures in order to have any hope of improving them. PMID- 16221523 TI - Is gestational age an independent variable affecting uterine scar rupture rates? AB - OBJECTIVE: To evaluate the influence of gestational age on uterine scar rupture. METHODS: This was a population-based study of data from Cardiff Births Survey over a 10-year (1990-1999) period. Women with only one previous lower segment caesarean section with singleton uncomplicated pregnancy of 37 or more week's gestation, undergoing trial of vaginal delivery were included. SPSS version 10 was used for statistical analysis. Mann-Whitney, Fisher's exact test and Chi square tests were used wherever appropriate. Odds ratio (OR) with confidence intervals (CI) was used to quantify the risk. Potential confounding by other factors was controlled using logistic regression and corrected odds ratios with 95% confidence intervals were calculated. The data was analysed separately for induced and spontaneous labours. Primary outcome measure assessed was uterine scar rupture rate. Secondary outcome measures were repeat caesarean section rates, maternal and perinatal mortality and morbidity. RESULTS: Total sample number was 1620. Eighty percent (n = 1301) of the population went into spontaneous labour and 20% (n = 319) were induced. Successful trial of vaginal birth was accomplished in 60% and trial of scar after estimated date of delivery did not alter this outcome significantly (39.1% versus 43.6%, p > 0.05). We noted an overall scar rupture rate of 0.9% (n = 14) and caesarean section rate of 40.4% (n = 654). Scar rupture rates significantly increased in women who underwent trial of labour after estimated date of delivery (p < 0.001, OR 6.3, CI 1.9-20.2) without a corresponding increase in caesarean section, maternal and perinatal morbidity figures. The influence of gestational age on scar rupture persisted even after controlling for other confounding factors such as birth weight, induction of labour and BMI (corrected OR 1.9, CI 1.1-3.5). CONCLUSIONS: The overall incidence of scar rupture and success of trial of scar after previous caesarean section in our population was similar to that quoted in the literature. Previous evidence has suggested that it is safe for these women to exceed 40 weeks gestation but our data do not support this. PMID- 16221524 TI - Impact of environmental chemicals on the thyroid hormone function in pituitary rat GH3 cells. AB - Endocrine disrupting chemicals (EDCs) are widespread in the environment and suspected to interfere with the function of thyroid hormones (THs). We investigated the TH disrupting activity of different classes of EDCs including plasticizers (bisphenol A, bisphenol A dimethacrylate), alkylphenols (4-n nonylphenol, 4-octylphenol), pesticides (prochloraz, iprodion, chlorpyrifos), PCB metabolites (OH-PCB 106, OH-PCB 121, OH-PCB 69) and brominated flame-retardants (tetrabromobisphenol A). The ED potential of a chemical was determined by its effect on the cell proliferation of TH-dependent rat pituitary GH3 cell line. All tested chemicals significantly interfered with the cell proliferation alone or upon co-treatment with T3. The growth of GH3 cells was stimulated by all tested chemicals, but 4-n-nonylphenol, 4-octylphenol, prochloraz and iprodion elicited an inhibitory effect on cell growth. In conclusion, these EDCs have the potential to exert TH disruption increasing the risk or a negative impact on fetal brain development, resulting in cognitive dysfunctions. PMID- 16221525 TI - Common fragile sites, extremely large genes, neural development and cancer. AB - Common fragile sites (CFSs) are large regions of profound genomic instability found in all individuals. They are biologically significant due to their role in a number of genomic alterations that are frequently found in many different types of cancer. The first CFS to be cloned and characterized was FRA3B, the most active CFS in the human genome. Instability within this region extends for over 4.0 Mbs and contained within the center of this CFS is the FHIT gene spanning 1.5 Mbs of genomic sequence. There are frequent deletions and other alterations within this gene in multiple tumor types and the protein encoded by this gene has been demonstrated to function as a tumor suppressor in vitro and in vivo. In spite of this, FHIT is not a traditional mutational target in cancer and many tumors have large intronic deletions without any exonic alterations. There are several other very large genes found within CFS regions including Parkin (1.37 Mbs in FRA6E), GRID2 (1.47 Mbs within 4q22.3), and WWOX (1.11 Mbs within FRA16D). These genes also appear to function as tumor suppressors but are not traditional mutational targets in cancer. Each of these genes is highly conserved and the regions spanning them are CFSs in mice. We have now examined lists of the largest human genes and found forty that span over one megabase. Many of these are derived from chromosomal bands containing CFSs. BACs within these genes are being utilized as FISH probes to determine if these are also CFS genes. Thus far we have identified the following as CFS genes: CNTNAP2 (2.3 Mbs in FRA7I), DMD (2.09 Mbs in FRAXC), LRP1B (1.9 Mbs in FRA2F), CTNNA3 (1.78 Mbs in FRA10D), DAB1 (1.55 Mbs in FRA1B), and IL1RAPL1 (1.36 Mbs in FRAXC). Although, these genes are also not traditional mutational targets in cancer they do exhibit loss of expression in multiple tumor types suggesting that they may also function as tumor suppressors. Many of the large CFS genes are involved in neurological development. Parkin is mutated in autosomal recessive juvenile Parkinsonism and deletions in mice are associated with the mouse mutant Quaking (viable). Spontaneous mouse mutants in GRID2 and DAB1 are associated with Lurcher and Reelin, respectively. In humans, alterations in IL1RAPL1 cause X-linked mental retardation and loss of WWOX is associated with Tau phosphorylation. We propose that the instability-induced alterations in these genes contribute to cancer development in a two-step process. Initial alterations will primarily occur within intronic regions, as these genes are greater than 99% intronic. These are not benign. Instead, they alter the repertoire of transcripts produced from these genes. As cancer progresses deletions will begin to encompass exons resulting in gene inactivation. These two types of alterations occurring in multiple large CFS genes may contribute significantly to the heterogeneity observed in cancer. There are also important potential linkages between normal neurological development and the development of cancer mediated by alterations in these genes. PMID- 16221526 TI - Ajoene inhibits both primary tumor growth and metastasis of B16/BL6 melanoma cells in C57BL/6 mice. AB - Ajoene is an organosulphur compound derived from garlic with important effects on several membrane-associated processes such as platelet aggregation, as well as being cytotoxic for tumor cell lines in vitro. In the present study, we investigated the effect of ajoene on different cell types in vitro, as well as its inhibitory effects on both primary tumors and metastasis in a mouse model. We found ajoene to inhibit tumor cell growth in vitro, but also to inhibit strongly metastasis to lung in the B16/BL6 melanoma tumor model in C57BL/6 mice. As far as we are aware, this is the first report of the anti-metastatic effect of ajoene. Ajoene also inhibited tumor-endothelial cell adhesion, as well as the in vivo TNF alpha response to lipopolysaccharide. Possible mechanisms of its antitumoral activity are discussed in the light of these results. PMID- 16221527 TI - Assessment of ethylene removal with Pseudomonas strains. AB - This study investigated the biological removal of ethylene by Pseudomonas strains in a batch test and a biofilter column. In the batch test, no removal of ethylene was found in the absence of inoculated system, whereas more than 50% of the ethylene in the presence of inoculated system was degraded within 17 h, and completely removed after 25 h. The biofilter, packed with activated carbons, was capable of achieving ethylene removal efficiency as much as 100% at a residence time of 14 min and an inlet concentration of 331 mg m-3. Under the same conditions, carbon dioxide with a concentration of up to 1097 mg m-3 was produced. It was found that carbon dioxide was produced at a rate of 87 mg day-1, which corresponded to a volume of 0.05 L day-1. During operation with an inlet ethylene of 331 mg m-3, the maximum elimination capacity of the biofilter was 34 g C2H4 m-3 day-1. This biological system could reduce the ethylene concentration to levels below the threshold limit for the plant hormonal response (0.01 mg m 3), and provide an attractive treatment technology in horticultural storage facilities. PMID- 16221528 TI - Degradation of aqueous solutions of camphor by heterogeneous photocatalysis. AB - In this study the photocatalytic degradation of aqueous solutions of camphor was investigated by using TiO2 and ZnO photocatalysts. In the presence of artificial UV-light the highly photosensitive camphor was almost totally degraded after reaction times of 60 min. However, under these conditions the mineralization degree was lower than 25%. In the presence of semiconductors the degradation was complete after a treatment time of about 30 min. Moreover, the mineralization was considerably greater, mainly with the use of TiO2 (> 80% at reaction time of 60 min). Heterogeneous photocatalytic processes applied in the presence of solar radiation show a promising degradation capability. TiO2-based processes afforded mineralization degrees of about 90% after a reaction time of 120 min, when the system was assisted by aeration. PMID- 16221529 TI - Use of hen feathers as potential adsorbent for the removal of a hazardous dye, Brilliant Blue FCF, from wastewater. AB - Waste material, hen feather, a biosorbent, was successfully utilized in removing a water-soluble hazardous triphenylmethane dye, Brilliant Blue FCF from wastewater. The paper incorporates effect of pH, temperature, amount of adsorbent, contact time, concentration of adsorbate, etc. The adsorption data validates Langmuir and Freundlich adsorption isotherms and on the basis of these isotherms thermodynamic parameters like Gibb's free energy, change in enthalpy and entropy were calculated. Kinetics of the ongoing adsorption was also monitored and specific rate constants for the involved process were calculated at different temperatures. Kinetic measurements suggest a first order adsorption kinetics and adsorption was found to be applicable via film diffusion process in the entire concentration range. PMID- 16221531 TI - The platelet maximum number of A2A-receptor binding sites (Bmax) linearly correlates with age at onset and CAG repeat expansion in Huntington's disease patients with predominant chorea. AB - Huntington's disease (HD) is caused by an expanded CAG mutation and may show a heterogeneous clinical presentation. To date, although the age at onset mostly depends on the expanded CAG repeat number, no validated easy-to-test biomarkers exist either for following up patients progression rate or for exactly predicting age at onset (defined as the time when motor clinical manifestations first became noticeable). We tested the function of A(2A) receptor, strongly expressed in the brain striatum and peripheral cells, in patients' blood platelets and confirmed a maximum number of binding sites (B(max)) higher than in controls (216 +/- 9 versus 137 +/- 7; p=0.0001). We found a linear correlation between the receptor B(max) and the expanded CAG repeat number (n=52, r(2)=0.19, p=0.0011). When we selected the patients according to their clinical presentation (according to the predominating motor manifestations) and plotted the receptor B(max) against patients' age at onset, we found a significant linear correlation only when considering those subjects with chorea predominant on all other motor symptoms (n=26, r(2)=0.39, p=0.0007). Because the typical chorea may depend on early dysfunction of the striatum in HD, peripheral A(2A) amplification in blood platelets might reflect a central dysfunction in this part of the brain. Further studies on a larger sample size should confirm whether the analysis of A(2A) receptor binding in patients' blood could be a useful clinical marker according to the patients' phenotype. PMID- 16221532 TI - Escherichia coli F4 fimbriae specific llama single-domain antibody fragments effectively inhibit bacterial adhesion in vitro but poorly protect against diarrhoea. AB - Oral administration of polyclonal antibodies directed against enterotoxigenic Escherichia coli (ETEC) F4 fimbriae is used to protect against piglet post weaning diarrhoea. For cost reasons, we aim to replace these polyclonal antibodies by recombinant llama single-domain antibody fragments (VHHs) that can be produced efficiently in microorganisms. Six F4 fimbriae specific VHHs were isolated. The VHH that was produced at the highest level by yeast, K609, was further analysed. 3.8 mg/L K609 inhibited 90% of bacterial attachment to intestinal brush borders in vitro. Perfusion of a jejunal segment with at least 4 mg/L K609 reduced the ETEC-induced fluid loss, but only to 30%. Preventive administration of a high K609 dose (150 mg/(piglet day)) to piglets that were challenge infected with ETEC resulted in less severe diarrhoea only at 4 and 5 days post-infection, but did not improve average daily weight gain, ETEC shedding and piglet survival. Thus, we have shown that an antibody fragment that effectively inhibited in vitro ETEC adhesion to intestinal brush borders poorly protected piglets against experimental ETEC infection. PMID- 16221533 TI - Effect of organophosphate pesticide diazinon on expression and activity of intestinal P-glycoprotein. AB - Organophosphate insecticide diazinon is widely used in agricultural practices, submitting farmers to repeated exposure. Because efflux pumps, as P-glycoprotein (P-gp), serve both as natural defense mechanisms and influence the bioavailability and disposition of drugs, we analyzed the ability of diazinon to act as efflux modulator. Oral administration of diazinon (2-20 mg/kg, 5 days, or 10 mg/kg, 2-12 days) increased intestinal mdr1a mRNA of rats, in both dose- and time-dependent manner, and increased the expression of intestinal P-gp. Using the intestinal cell-line Caco-2, we found that 100 microM diazinon significantly inhibited digoxin and vinblastine secretive flux through the cell monolayers, whereas digoxin and vinblastine absorptive flux increased. The 25 microM diazinon was transported preferentially in basolateral (BL) to apical (AP) direction, suggesting a net secretion. The efflux rate significantly decreased in the presence of metabolic inhibitors sodium azide and 2-deoxy-d-glucose, P-gp inhibitors cyclosporin A and valspodar, but not in the presence of MRPs inhibitor MK571. Repeated exposure of Caco-2 cells to diazinon increased P-glycoprotein expression and activity. These results suggested the involvement of P-gp in the transfer of diazinon, leading to potential consequences for xenobiotic interactions, and showed that repeated exposure to low doses of pesticide may lead to up-regulated P-gp functions in the intestine of mammals. PMID- 16221534 TI - Pomegranate as a cosmeceutical source: pomegranate fractions promote proliferation and procollagen synthesis and inhibit matrix metalloproteinase-1 production in human skin cells. AB - Pomegranate (Punica granatum) is an ancient fruit with exceptionally rich ethnomedical applications. The peel (pericarp) is well regarded for its astringent properties; the seeds for conferring invulnerability in combat and stimulating beauty and fertility. Here, aqueous fractions prepared from the fruit's peel and fermented juice and lipophilic fractions prepared from pomegranate seeds were examined for effects on human epidermal keratinocyte and human dermal fibroblast function. Pomegranate seed oil, but not aqueous extracts of fermented juice, peel or seed cake, was shown to stimulate keratinocyte proliferation in monolayer culture. In parallel, a mild thickening of the epidermis (without the loss of ordered differentiation) was observed in skin organ culture. The same pomegranate seed oil that stimulated keratinocyte proliferation was without effect on fibroblast function. In contrast, pomegranate peel extract (and to a lesser extent, both the fermented juice and seed cake extracts) stimulated type I procollagen synthesis and inhibited matrix metalloproteinase-1 (MMP-1; interstitial collagenase) production by dermal fibroblasts, but had no growth-supporting effect on keratinocytes. These results suggest heuristic potential of pomegranate fractions for facilitating skin repair in a polar manner, namely aqueous extracts (especially of pomegranate peel) promoting regeneration of dermis, and pomegranate seed oil promoting regeneration of epidermis. PMID- 16221535 TI - Application of false discovery rate procedure to pairwise comparisons of refractive index of glass fragments. AB - This study was undertaken to apply a new method of controlling type I error when performing pairwise comparisons. The Benjamini and Hochberg false discovery rate (FDR) controlling procedures have proved to be very powerful tools in solving many practical problems but have not yet been applied to pairwise comparisons of refractive index of glass samples. Students t-test and Welch test (unequal variance Student's t-test) were applied to all possible pairwise comparisons. The comparisons were made on the basis of refractive index values of 72 glass fragments from different car windows and 69 different fragments from one windscreen. The type I error was controlled by the use of Benjamini and Hochberg false discovery rate (FDR) controlling procedure. To illustrate the importance of controlling type I error when using pairwise comparisons, results of pairwise comparisons with the FDR controlling procedure were compared to those comparisons made without any controlling procedure. Significantly fewer false negative results (false rejection of H0) were found during the use of FDR procedure in the comparison of refractive indices from the same windscreen than when comparing without controlling type I error. The results of application of FDR method were also compared to the results of other post-hoc tests, such as Tukey HSD test and Bonfferoni test. The FDR method has higher power than Bonferroni method and Tukey HSD method and control errors better than comparing without controlling type I error. The method of choice, for pairwise comparison of glass fragments on the basis of refractive index is Welch test with FDR adjustment. The method gave type I errors at level about 5.3% and type II error at about 3.7%. PMID- 16221536 TI - The effects of larval crowding and food type on the size and development of the blowfly, Calliphora vomitoria. AB - The use of entomological evidence in the estimation of the post mortem interval (PMI) often depends on the size and developmental stage of blowfly larvae collected from a corpse. Therefore, factors which can have an effect on the larval size and growth rate can have implications for reliable PMI determinations. This study explores the competitive effects of larval overcrowding on Calliphora vomitoria reared on three different pig tissues- liver, brain and muscle. The competitive feeding environment within the more crowded larval cultures resulted in increased development rates and the production of undersized larvae and adults. Variation in the extent of these effects was observed on each of the three body tissues, highlighting the importance of documenting the positions from which entomological evidence is recovered from a corpse. PMID- 16221537 TI - Genetic data on 10 STR loci a population of western Poland. AB - Allele frequencies for 10 STRs included in the AmpFlSTR SGM Plus kit were determined in a population sample of 668 unrelated individuals living in western Poland. All loci met Hardy-Weinberg expectations. Exact tests disequilibrium analysis revealed two departures from independence out of 45 pairwise comparisons. The combined matching probability (MP) and power of exclusion (PE) for all 15 loci are 2.56 x 10(-13) and 0.99996, respectively. PMID- 16221538 TI - Alternate metabolism during the dauer stage of the nematode Caenorhabditis elegans. AB - When environmental conditions are unsuitable to support nematode reproduction, Caenorhabditis elegans arrests development before the onset of sexual maturity and specialised 'dauer' larvae, adapted for dispersal, and extended diapause are formed. Dauer larvae do not feed and their metabolism is dependent on internal food reserves. Adult worms which express defects in the insulin/insulin-like growth factor receptor DAF-2 also display enhanced longevity. Whole genome mRNA expression profiling has demonstrated that C. elegans dauer larvae and daf-2 adults have similar transcription profiles for a cohort of longevity genes. Important components of this enhanced longevity system are the alpha-crystallin family of small heat shock proteins, anti-ROS defence systems, increased activity of cellular detoxification processes and possibly also increased chromatin stability and decreased protein turnover. Anaerobic fermentation pathways are upregulated in dauer larvae, while long-lived daf-2 adults appear to have normal oxidative metabolism. Anabolic pathways are down regulated in dauer larvae (and possibly in daf-2 adults as well), and energy consumption appears to be diverted to enhanced cellular maintenance and detoxification processes in both systems. PMID- 16221539 TI - The effect of melatonin treatment on the ovarian response of ewes to the ram effect. AB - To determine the ovarian response to the ram effect after treatment with melatonin, on 8 March, 71 Rasa Aragonesa ewes were randomly assigned to either the treatment group and given an 18mg melatonin implant or the untreated group. On 19 April (day 0), rams were introduced into the flock. Melatonin treatment produced a significantly higher percentage of cyclic ewes at ram introduction (P<0.05). Melatonin-treated ewes had their first oestrus after ram introduction significantly earlier than did untreated ewes (P<0.0001), and the groups differed in the distribution of their ovarian response. Most (80%) of the treated ewes exhibited a silent ovulation followed by a cycle of normal duration, whereas about half (52%) of the untreated ewes exhibited a silent ovulation, a short cycle, and another silent ovulation followed by a cycle of normal duration (P<0.05). At ram introduction, melatonin-treated ewes, cyclic and non-cyclic, had higher mean plasma progesterone concentrations than did untreated ewes. The proportion of ewes that mated within the first 17 days of the mating period was significantly higher among the treated ewes than in the untreated ewes (P<0.0001). Furthermore, at lambing, 39% of the melatonin-treated ewes lambed within the first 17 days of the lambing period, while none of the untreated ewes lambed in that period. The untreated group exhibited peaks in mating between days 18 and 21, and particularly, between days 22 and 25, when the majority of ewes lambed; peaks did not occur in the treatment group. Treated and untreated ewes did not differ significantly in fertility, litter size and fecundity. In conclusion, melatonin treatment modifies the ovarian response to the ram effect in ewes, which leads to modifications in mating patterns, and consequently, the lambing curve. PMID- 16221540 TI - Vancomycin versus teicoplanin in the therapy of experimental methicillin resistant Staphylococcus aureus (MRSA) meningitis. AB - The aim of this study was to compare the antibacterial activity of teicoplanin and vancomycin in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) meningitis using a rabbit meningitis model. The MRSA strain ATCC 43300 was used to infect the rabbits. The vancomycin group received 20 mg/kg vancomycin every 12h (q12h), the teicoplanin group received 6 mg/kg teicoplanin q12h and the control group did not receive any treatment. Drug levels were measured using a bioassay technique. Bacterial counts in the treatment groups were significantly lower (P<0.05) than those of the control group at 12 h and 24 h after treatment. When the treatment groups were compared, the bacterial counts after 12 h or 24 h of treatment were similar (P>0.05). These data suggest that the antibacterial activity of vancomycin and teicoplanin are similar in experimental MRSA meningitis of rabbits. PMID- 16221541 TI - Community consumption of antibacterial drugs within the Jordanian population: sources, patterns and appropriateness. AB - This study is the first of its type to evaluate sources, patterns and appropriateness of antibacterial drug consumption within the Jordanian population. It uses a structured random interview to customers arriving at community pharmacy stores seeking antibacterial medication for systemic infections. Dispensed antibacterial drugs belonged to eight different main classes of systemically used antibacterial drugs in accordance with the Anatomical Therapeutic Chemical (ATC) classification system. Just less than one half of all dispensed antibacterial drugs were without a prescription (46%), either via self-medication (23.2%) or pharmacist recommendation (23.1%). Inappropriateness in use was seen in 29.9% and 34% of the prescribed and non prescribed (over-the-counter) antibacterial drugs, respectively. No uniformity in the treatment profile of the different treated complaints was seen. Our study shows great misuse and abuse of antibacterial drugs and hence there is a need to force regulations to control community use of antibacterial drugs and to increase awareness of the consequences of their inappropriate and uncontrolled use. PMID- 16221542 TI - "Cooking the sample": radiofrequency induced heating during solid-state NMR experiments. AB - Dissipation of radiofrequency (RF) energy as heat during continuous wave decoupling in solid-state NMR experiment was examined outside the conventional realm of such phenomena. A significant temperature increase could occur while performing dynamic NMR measurements provided the sample contains polar molecules and the sequence calls for relatively long applications of RF power. It was shown that the methyl flip motion in dimethylsulfone (DMS) is activated by the decoupling RF energy conversion to heat during a CODEX pulse sequence. This introduced a significant bias in the correlation time-temperature dependency measurement used to obtain the activation energy of the motion. By investigating the dependency of the temperature increase in hydrated lead nitrate on experimental parameters during high-power decoupling one-pulse experiments, the mechanisms for the RF energy deposition was identified. The samples were heated due to dissipation of the energy absorbed by dielectric losses, a phenomenon commonly known as "microwave" heating. It was thus established that during solid state NMR experiments at moderate B0 fields, RF heating could lead to the heating of samples containing polar molecules such as hydrated polymers and inorganic solids. In particular, this could result in systematic errors for slow dynamics measurements by solid-state NMR. PMID- 16221543 TI - Trunk muscle co-ordination during gait: relationship between muscle function and acute low back pain. AB - Low back pain costs billions of Euros annually in all industrialized countries. Often radiological diagnosis fails to give evidence of the pathogenesis of low back pain. Although psychophysiological characteristics have an influence, it seems that insufficient muscular spinal stabilization may play the major role in the development of low back pain. Assessment of trunk muscle stabilization activity during everyday activities is rare. Therefore, in this study healthy persons were investigated during walking on a treadmill at a speed of 4 km/h. Women (n = 16) with no history of back pain were investigated before and after a static loading situation of the spine, i.e. while wearing a waistcoat. After this loading situation four women developed pain (pain subjects). Surface EMG (SEMG) was taken from five trunk muscles of both sides. Grand averaged amplitude curves over stride, amplitude normalized curves and variation between all included strides were calculated for all muscles and subjects, respectively. The normal range of all calculated parameters was defined within the span between the 5th and the 95th percentiles of all pain free subjects. Data were evaluated according to deviations from the normal range. Already before the load situation, pain subjects showed considerable deviations from the normal range, mainly of their abdominal muscles. There was no relationship between magnitude of deviation and pain intensity, but perceived exertion was highest in those subjects who showed the most symptoms in terms of number of muscles being identified as considerably deviating from the normal range. No specific "dysfunction pattern" could be identified, which argues for highly individual mechanisms instead of a single target muscle. The results suggest cumulative effects of different disturbance levels resulting in acute back pain. Since deviations could be identified already before the pain occurred, disturbed muscle function seems to be a risk factor for developing back pain. Further investigations aimed at clear identification of and, as a second step, correction of muscle function are necessary. PMID- 16221544 TI - Where disease pathogenesis meets protein formulation: renal deposition of immunoglobulin aggregates. AB - Aggregation is one of the important issues encountered during the development of immunoglobulin-based drugs. The aim of the current review is to discuss the causes and consequences of immunoglobulin aggregation as well as the relevance of immunoglobulin aggregation to disease pathogenesis. Extracellular deposition of immunoglobulins, either monoclonal light chains or intact polyclonal antibodies, induces renal failure in various nephropathies. The aggregates can present fibrillar or amorphous structures. In this review, factors known to influence protein aggregation, such as the primary structure of the protein, local environment and glycosylation are assessed, as well as the subsequent altered clearance, fibril formation and toxicity. The role of the protein local environment is emphasized. Even if the local environment causes only minor perturbations in the protein structure, these perturbations might be sufficient to trigger aggregate formation. This fact underlines the importance of choosing appropriate formulations for protein drugs. If the formulation provides a slightly destabilizing environment to the protein, the long-term stability of the drug may be compromised by aggregate formation. PMID- 16221545 TI - Changes in the amide I FT-IR bands of poly-L-lysine on spray-drying from alpha helix, beta-sheet or random coil conformations. AB - Poly-L-lysine (PLS) was spray-dried in a laboratory-scale, mini spray-dryer at Tin/Tout =150/90-95 degrees C from three different liquid feeds composed mainly of alpha-helix, beta-sheet or random coil conformations of the homopolypeptide. FT-IR analysis of the liquid feeds, the spray-dried solids, and the re-dissolved solids was performed by considering the deconvoluted and second-derivative amide I spectra, as well as a Gaussian curve fitting procedure. All three initial conformations were transformed by spray-drying to anti-parallel beta-sheet with bands at 1623 and 1690 cm(-1). The beta-sheet liquid feed showed a band at 1616 cm(-1) indicating a denatured, extended chain structure that was also converted to anti-parallel beta-sheet on spray-drying. The shift to beta-sheet cannot therefore be a simple result of forming the conformation with the strongest H bonds in the dried state. We suggest that steric effects arising from the close approach of the globular polypeptide molecules during drying make the anti parallel beta-sheet structure energetically favorable in the solid state. This suggestion is supported by the effects of trehalose on the FT-IR amide I bands of the spray-dried PLS. No stabilizing effects were observed on either the initial alpha-helix or beta-sheet (extended chain) conformations. Random coil could be partially stabilized. Again, no direct relation to H-bond strength is evident. The efficacy of the trehalose could be related to the ability of newly-formed trehalose/PLS intermolecular H-bonds to stabilize the intramolecular H-bonds of the secondary structural elements. PMID- 16221546 TI - Dietary eicosapentaenoic acid modulates CTLA-4 expression in murine CD4+ T-cells. AB - We have demonstrated that downregulation of proliferation by CD4(+) T-cells in mice fed n-3 PUFA diets is dependent on the involvement of CD28. Therefore, we hypothesized that the balance of co-stimulatory and downregulatory properties of CD28 and CTLA-4, respectively, would be altered by diet. Mice were fed a control corn oil (CO)-enriched diet devoid of n-3 PUFA, or diets enriched with either docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA) for 14d. The proliferation of splenic CD4(+) T-cells was suppressed by DHA and EPA following stimulation with anti-CD3 and anti-CD28. Surprisingly, the number of surface CD28 molecules was not reduced in activated CD4(+) T-cells from either group of n-3 PUFA-fed mice. However, in mice fed EPA, CTLA-4 protein levels were enhanced significantly 72 h post-activation (P<0.01). Therefore, we conclude that dietary EPA may suppress CD4(+) T-cell activation by enhancing the downregulatory co receptor CTLA-4, while not altering the levels of CD28. PMID- 16221547 TI - A new questionnaire specifically designed for patients affected by chronic obstructive pulmonary disease; The Italian Health Status Questionnaire. AB - The aim of this study was to develop a specific and valid questionnaire for Italian COPD patients, living on the north or the south of Italy-which are two culturally distinct areas. The project consisted in three steps: (1) initial item set generation to identify items relevant to both genders, all ages and both regions; (2) item reduction including tests of regional specificity; (3) tests of internal validity using item-response theory using Rasch one-parameter modelling. Ninty-six COPD patients (mean aged 69 yr; 78 Male) completed the original set of 124 items of the Italian Health Status Questionnaire (IHSQ). Item reduction was carried out using an established standardised approach employing classical psychometric test theory. The internal construct validity of the 47 items that survived this process were tested to determine whether they constituted a unidimensional construct "impaired health due to COPD" using Rasch analysis. This showed that the questionnaire had very good psychometric properties, with an excellent Person Separation Index of 0.95 and no evidence of bias due to item trait interaction (chi104(2)=127.1, P=n.s.). The combination of classical test theory and modern item-response methodology has produced a questionnaire with excellent measurement properties suitable for COPD patients whether from the north or south of Italy. PMID- 16221548 TI - Effects of anti-asthma therapy on dyspnea perception in acute asthma patients. AB - Blunted perception of dyspnea may predispose patients to fatal asthma attacks. To examine whether this impaired perception of dyspnea in patients with acute asthma could be corrected by anti-asthma therapy, the medical records of 104 consecutive asthma patients who had been hospitalized as a result of asthma attacks were analyzed retrospectively. During the course of treatment with conventional asthma medications, the forced expiratory volume in 1s (FEV1) and the Borg scale-based dyspnea perception scores during breathing through an inspiratory muscle trainer were measured at least twice. The baseline Borg score measured just before discharge was significantly lower than from that measured initially, regardless of improvement in FEV1. In contrast, the Borg score at the highest resistance (HR; 3.12+/-0.26 vs. 5.03+/-0.53; P<0.01) and the HR-induced DeltaBorg score (1.68+/-0.20 vs. 4.47+/-0.54, P<0.001) were increased significantly in the Poor Perceivers (Borg score 5 at HR and HR-induced DeltaBorg score 3). Patient age (r=0.363, P<0.001), blood eosinophil counts (r=-0.285, P<0.01), and serum total IgE levels (r=-0.213, P<0.05), but not FEV1, were significantly related to the effect of the treatment on the HR-induced DeltaBorg scores. These findings suggest that anti-asthma treatments decrease dyspnea even without a concomitant improvement in lung function and correct the impaired perception of inspiratory resistive load in acute asthma, and that age and allergy influence the effect of treatment on impaired perception. PMID- 16221549 TI - Adaptations to normal human gait on potentially slippery surfaces: the effects of awareness and prior slip experience. AB - Prior knowledge of potentially slippery conditions has been shown to alter normal human gait in slip and fall experiments. Here we quantify the effects of two aspects of prior knowledge - awareness of a possible slip and prior slip experience - on normal gait. Sixty-eight subjects (40F, 28M) each walked over 48 high-friction surfaces (control trials) and 12 low-friction surfaces. Within- and between-subject changes in lower limb muscle activation, gait kinematics and ground reaction forces were analyzed in three non-slip control trials: one before and one after the first unexpected slip exposure, and a third after repeated slip exposures. Subjects knew they might slip in the latter two trials but not the first trial. Twenty subjects slipped during their first low-friction exposure (early slip group), 32 in later low-friction exposures (late slip group), and 16 subjects did not slip at all. Simultaneous changes in awareness and experience between the first two analyzed trials of the early slip group altered the muscle activity in both limbs, reduced the foot and knee angles at heel strike in the slip limb and reduced the ground reaction forces, impulses and utilized friction after heel strike in the slip limb. A change in only awareness between the first two analyzed trials of the late slip group produced the same kinematic changes seen in the early slip group, but only small muscle activity change and no kinetic changes. Subsequent slip experience in the late slip group produced the muscle activation and kinetic changes observed in the early slip group, but no further kinematic changes. These results showed that awareness of a potential slip primarily alters how the slip-limb approaches the floor, whereas prior slip experience primarily alters the anticipatory muscle activation and how the foot interacts with the floor. These muscle, kinematic and kinetic changes were consistent with a more cautious "normal" gait, and can reduce the external validity of slip and fall experiments. PMID- 16221551 TI - Pharmacokinetic interaction of tetracycline with non-steroidal anti-inflammatory drugs via organic anion transporters in rats. AB - The present study aims to investigate the pharmacokinetic interaction between non steroidal anti-inflammatory drugs and tetracycline in rats. Pharmacokinetic parameters were determined following an intravenous administration of tetracycline (5 mg kg(-1)) to rats in the presence and absence of naproxen or diclofenac (20 mg kg(-1)). Compared to the control (given tetracycline alone), pretreatment with naproxen or diclofenac 30 min prior to tetracycline administration significantly altered the pharmacokinetics of tetracycline. Renal clearance of tetracycline was reduced by approximately three-fold in the presence of naproxen or diclofenac. Consequently, the systemic exposures (AUC) of tetracycline in the rats pretreated with naproxen or diclofenac were significantly (p < 0.05) higher than those from the control group given tetracycline alone. Furthermore, mean terminal plasma half-life of tetracycline was enhanced by two- to five-folds under the pretreatment with naproxen or diclofenac. Those results suggest that NSAIDs such as naproxen and diclofenac are effective to alter the renal elimination and pharmacokinetic profiles of tetracycline. Therefore, concomitant use of naproxen or diclofenac with tetracycline may require close monitoring for clinical consequence of potential drug interaction. PMID- 16221550 TI - Minimal versus conventional cardiopulmonary bypass: assessment of intraoperative myocardial damage in coronary bypass surgery. AB - OBJECTIVE: Minimal extracorporeal circulation (mini-ECC) is a new technology, consisting of a centrifugal pump, an oxygenator, and a modified suction system. The main advantage of mini-ECC is the reduction of tubing length (reduction of the priming volume). Additional beneficial effects are a decrease of coagulation cascade and a reduction of blood transfusion in patients undergoing coronary artery bypass grafting (CABG) surgery. We compared the intraoperative and early postoperative myocardial damage and outcome of patients who underwent CABG surgery with conventional cardiopulmonary bypass (CPB) or mini-ECC. METHODS: One hundred and thirty-six consecutive patients who underwent isolated CABG surgery at our institution were prospectively studied. Fifty-four patients (39.7%) were operated with mini-ECC. Patient characteristics were similar in both groups. The most interesting intraoperative details as well as in-hospital outcome were assessed. RESULTS: There was no difference in mortality between the two groups. Cross-clamping time was similar in both groups (p = 0.07). Defibrillation was required in one patient in the mini-ECC group (1.9%) and in 38 patients (46.3%) in the CPB group (p < 0.001). In the mini-ECC group, the requirement of inotropic support and incidence of atrial fibrillation was significantly lower than in the CPB group. Postoperative creatine kinase isoenzyme MB (CK-MB) and cardiac Troponin I (cTnI) were significantly lower in the mini-ECC group (p < 0.05). Duration of ventilation, length of stay in the intensive care unit and total hospitalization time were significantly shorter in patients operated with mini ECC (p < 0.05). CONCLUSION: Mini-ECC is a safe procedure and is followed by a diminished release of CK-MB and cTnI than after CPB. Postoperative recovery is accelerated following mini-ECC and there is a significantly lower incidence of postoperative atrial fibrillation. PMID- 16221552 TI - Tracking thymocyte migration in situ. AB - The dynamic process of thymocyte migration can now be visualized in real-time and in the context of the native thymic environment. With improved computational resources, key information can be extracted from real-time imaging data and the migratory behaviors of developing thymocytes can be quantitated. The extraction and exploitation of three dimensional data through time is providing new insight into the nature and regulation of intrathymic migration. In this review we discuss this interdisciplinary approach and the promise it holds for the study of thymocyte migration in situ. PMID- 16221553 TI - Clustering of domains of functionally related enzymes in the interaction database PRECISE by the generation of primary sequence patterns. AB - The PRECISE database was developed by our laboratory to allow for the systematic study of the ligand interactions common to a set of functionally related enzymes, where an interaction site is defined broadly as any residue(s) that interact with a ligand. During the construction of PRECISE, enzyme chains are extracted from the protein data bank (PDB) and clustered according to functional homology as defined by the enzyme commission (EC) nomenclature system. A sequence representative is chosen from each cluster based on the criterion set forth by the non-redundant PDB set, and pair-wise alignments of each cluster member to the representative are performed. Atom-based residue-ligand interactions are calculated for each cluster member, and the summation of ligand interactions for all cluster members at each aligned position is determined. Although we were able to successfully align most clusters using a simple dynamic programming algorithm, several cluster created exhibited poor pair-wise alignments of each cluster member to its sequence representative. We hypothesized that the observed alignment problems were, in most cases, due to the incorrect separation and alignment of different domains in multi-domain proteins, a mistake that frequently causes error proliferation in functional annotation. Here we present the results of generating primary sequence patterns for each poorly aligned cluster in PRECISE to assess the extent to which multi-domain proteins that are incorrectly aligned contributes to poor pair-wise alignments of each cluster member to its representative. This requires the use of an iterative locally optimal pair-wise alignment algorithm to build a hierarchical similarity-based sequence pattern for a set of functionally related enzymes. Our results show that poor alignments in PRECISE are caused most frequently by the misalignment of multi-domain proteins, and that the generation of primary sequence patterns for the assignment of sequence family membership yields better alignments for the functionally related enzyme clusters in PRECISE than our original alignment algorithm. PMID- 16221554 TI - Age-dependent increase of clusterin in the human pituitary gland. AB - Clusterin is a glycoprotein known to play various physiological roles including complement activity, amyloid binding activity in Alzheimer disease, as well as binding with heat shock proteins and abnormal prions. The present study immunohistochemically investigated the expression of clusterin in the human pituitary gland in subjects of 10-88 years of age (n=173). Causes of death were blunt injury (n=35), sharp injury (n=15), poisoning (n=11), drowning (n=14), fire fatalities (n=28), asphyxiation (n=15), hypothermia (n=7), hyperthermia (n=3), and natural diseases (n=45). Clusterin was detected in mixed cell follicles and the anterior lobar parenchymal cells. The area occupied by cells positive for clusterin were measured, and the ratio to the whole area of the anterior lobe (% clusterin-positive cell area) was estimated. There was a good correlation between the age of the subjects in years and the % clusterin-positive cell area in the anterior lobe of the pituitary gland (r=0.736, P<0.01). Relationships between % clusterin-positive cell and gender, cause of death, and survival time were insignificant. These findings indicate an age-dependent accumulation of clusterin in the pituitary gland, which may be related to the aging of endocrine systems. PMID- 16221555 TI - Y-chromosome STR haplotype profiling in the Mongolian population. AB - Eleven Y-chromosome STR loci (DYS19, DYS388, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, two DYS385 loci and DXYS156Y) were analyzed in 92 randomly chosen males from Mongolia. Allelic frequencies and gene diversity for each Y-STR locus and haplotype diversity were determined to evaluate their usefulness in forensic casework. A total of 84 different haplotypes were identified, among which 77 (83.7%) were individual-specific. The most frequent haplotype surveyed here was found in three individuals (3.6%), of which the 'minimal haplotype' (excluding DYS388 and DXYS156Y) was not found in the worldwide Y-STR haplotype reference database (YHRD) available at . PMID- 16221556 TI - Postmortem stability of pituitary hormones in the human adenohypophysis. AB - The hypophysis is embedded in the fossa at the base of skull, having important functions in the hormonal system. The present study investigated its postmortem morphological changes and the stability of adenohypopyseal hormones. The pituitaries were collected at autopsy 6 h to 20 days postmortem and were studied by histology, immunohistochemistry and electron microscopy. To avoid the influence of prolonged brain hypoxia or swelling, subjects who survived not longer than 12 h were examined. Histological changes were seen in the nucleus 6 h after death, followed by cytoplasmic changes, and the cell shapes were hardly identifiable 7 days postmortem. Electron microscopy revealed evident ultra structural changes 6 h postmortem, involving rough endoplasmic reticulum, Golgi complexes, mitochondria, nuclei and cell membranes. However, secretory granules remained well preserved 7 days postmortem. Immunostaining showed positivities for growth hormone, prolactin, adenocorticotropic hormone, luteinizing hormone and thyroid-stimulating hormone up to 15 days after death. These findings suggest the usefulness of immunohistochemical investigation of the adenohypophysis for estimating the time of death and endocrinologic evaluation in decomposed cadavers. PMID- 16221558 TI - Application of FTA technology to extraction of sperm DNA from mixed body fluids containing semen. AB - FTA technology is a novel method designed to simplify the collection, shipment, archiving and purification of nucleic acids from a wide variety of biological sources. In this study, we report a rapid and simple method of extracting DNA from sperm when body fluids mixed with semen were collected using FTA cards. After proteinase K digestion of the sperm and body fluid mixture, the washed pellet suspension as the sperm fraction and the concentrated supernatant as the epithelial cell fraction were respectively applied to FTA cards containing DTT. The FTA cards were dried, then directly added to a polymerase chain reaction (PCR) mix and processed by PCR. The time required from separation of the mixed fluid into sperm and epithelial origin DNA extractions was only about 2.5-3h. Furthermore, the procedure was extremely simple. It is considered that our designed DNA extraction procedure using an FTA card is available for application to routine work. PMID- 16221557 TI - Two fatal cases of child abuse in which neighbors were unaware of the victims' disappearance for a long period. AB - We describe two fatal cases of child abuse in which neighbors were unaware of the victims' disappearance for 1.5 months and 1 year, respectively. Recently, there have been fatal child abuse cases in which neighbors have been aware of the signs of the abuse but not notified the Child Care Authorities. Lack of concern about child welfare in the community is the greatest obstacle to protecting children at risk of abuse. The most effective means of preventing child abuse is to educate the community about how to recognize the signs of abuse and to inform the authorities. We emphasize that the community has an obligation to protect children against crime, including child abuse. The roles of the Social Services in preventing child abuse have been extended. Forensic pathologists are required to play a key role in child abuse prevention, and in Japan their activity should be extended to the administrative field. PMID- 16221559 TI - Where not to live: a geo-demographic classification of mortality for England and Wales, 1981-2000. AB - The aim of this paper is to pilot a method for geo-demographic classification for mortality patterns in Britain. Age and sex directly standardised mortality ratios (DSMRs) for 100 grouped International Classification of Disease series 9 causes of death (ICD-9) were calculated. The 84 European Parliamentary (EP) constituencies as defined in 1999 were used as the spatial basis of this study to allow regional comparisons to be made while comparing units of similar population sizes. Scotland was excluded from the final analysis, leaving 76 regions. This paper is a preliminary investigation of the patterns of the causes of death over time and space in England and Wales using cluster analysis to summarise some of the structure in the data. Seven major and three minor cluster profiles were developed. PMID- 16221560 TI - Turns amplitude analysis of the orbicularis oculi and oris muscles. AB - OBJECTIVE: The aim of the study was to determine whether 'clouds' from turns amplitude analysis obtained from the orbicularis oculi and oris muscles without force monitoring can be used to differentiate pathological processes affecting the face. METHODS: The interference pattern from orbicularis oculi and orbicularis oris was studied using a concentric needle electrode. Data-points from 20 normal subjects were plotted on a logarithmic scale of mean amplitude between turns versus turns/second, from which linear regression analysis defined the 95% confidence intervals. This enabled us to draw the boundaries of the normal cloud on a linear plot. Data-points from the interference pattern in two pathological cohorts, of 6 patients receiving botulinum toxin injections (representing a neurogenic model), and 6 patients with a muscle dystrophy (representing a myopathic model) were plotted against the normal cloud. These findings were compared and correlated with the mean durations obtained on motor unit action potential analysis from these same two facial muscles. RESULTS: The majority of patients receiving botulinum toxin injections into their facial muscles showed a pattern of high amplitude with low turns/s, or low amplitude with a low-to-normal range of turns/s in both facial muscles. These findings were associated with high-duration motor unit action potentials in most cases. In the myopathic group of patients 66% showed a pattern of low amplitude with low-to normal range of turns/s in O oculi and O oris. This correlated with short duration motor unit action potentials in both facial muscles. CONCLUSIONS: We have demonstrated that turns amplitude analysis without force monitoring can be used to study the interference pattern from facial muscles and can be applied to differentiate primary neurogenic from myopathic pathological processes. SIGNIFICANCE: Turns amplitude analysis without force monitoring in the facial muscles can be used as an effective and practical method of interference pattern analysis to complement findings from conventional motor unit action potential analysis. PMID- 16221561 TI - Reduced sensorimotor inhibition in the ipsilesional motor cortex in a patient with chronic stroke of the paramedian thalamus. AB - OBJECTIVE: Unilateral or bilateral paramedian infarction in the region of the thalamus and upper midbrain may lead to hypersomnia. To determine whether unilateral infarction of the paramedian thalamus leads to changes in excitability of ipsilesional primary motor hand area (M1). METHODS: We describe a patient with chronic stroke of the right dorsomedian and intralaminar thalamic nuclei, who suffered from mild persistent hypersomnia. We studied the excitability of the right and left M1 with transcranial magnetic stimulation (TMS) in the patient, and in 10 healthy controls. RESULTS: In contrast to healthy controls, contralateral electrical stimulation of the median nerve failed to induce short latency afferent inhibition (SAI) in the ipsilesional M1. Other measures of corticomotor excitability and somatosensory evoked potentials were normal. CONCLUSIONS: The selective loss of ipsilateral SAI in a patient with paramedian thalamic stroke suggests that during wakefulness, the intact paramedian thalamus facilitates the excitability of intracortical inhibitory circuits, which process thalamocortical sensory inputs in the ipsilateral M1. This preliminary finding suggests that measurements of SAI may provide a means of probing the integrity of some neural pathways, which are involved in the control of wakefulness and arousal. SIGNIFICANCE: In addition to the established role of the paramedian thalamus in arousal and memory, our observation suggests that thalamocortical projections from the paramedian thalamus contribute to the integration of sensory input at the cortical level during wakefulness. PMID- 16221562 TI - Lower frequency variability in the alpha activity in EEG among patients with epilepsy. AB - OBJECTIVE: Clinically, intra-subject variability of the alpha frequency is well known, but sparsely documented and practically not used in the evaluation of the electroencephalogram (EEG). In cognitive science, however, the peak alpha frequency (PAF) variations are implemented. The aim of the present study was to document the clinical notion of differences in alpha frequency variations in patients with epilepsy compared to a control group. METHODS: Standard EEG recordings from 28 patients, 18 with epilepsy and 10 patients having EEG for other reasons were included. Ten seconds of artifact free EEG were sampled from F3, F4, T5, T6, O1 and O2 at the beginning, after hyperventilation and at the end of a 20 m recording and Fast Fourier transforms was applied to these epochs of each recording. RESULTS: The study showed a lower frequency in the epilepsy group (frontal 9.22 vs. 10.24 Hz, temporal 9.18 vs. 9.88 Hz and occipital 9.42 vs. 10.30 Hz) and lower frequency variability, with lower values in the epilepsy group in occipital (0.8 vs. 1.44 Hz) and temporal leads (0.89 vs. 1.39 Hz). Frontally, the variability was not significant (0.71 vs. 1.18 Hz, P = 0.0824). Within the groups, there was no relation between frequency and variability. CONCLUSIONS: This study shows that there is PAF variability in the alpha activity. This variability is compromised in patients with epilepsy. Lower alpha frequency is observed in epilepsy group. It is to some extent due to antiepileptic drugs. The lower alpha frequency variability is probably due to a different mechanism as there is no relation between the frequency and its variability within the two groups. SIGNIFICANCE: The alpha activity shows physiological frequency variations that may be compromised by epilepsy. PMID- 16221563 TI - Reference values of fractionated neurography of the ulnar nerve at the wrist in healthy subjects. AB - OBJECTIVE: This study is designed to derive a normative database for nerve conduction values of the ulnar nerve in the wrist. METHODS: Ulnar nerve study at the wrist (UNSW) was performed in 204 hands of 102 control subjects. The UNSW was composed of motor and sensory tests. Motor UNSW was done with first dorsal interosseous muscle recording. Sensory UNSW was performed antidromically with fifth finger recording. The 3 stimulation points were 2 cm proximal to the pisiform, just lateral to pisiform, and 3 cm distal to the pisiform. RESULTS: Mean latency differences in the proximal and distal segments were 0.4 +/- 0.1 and 0.5 +/- 0.1 ms in motor UNSW and 0.4 +/ -0.1 and 0.5 +/- 0.1 ms in sensory UNSW. The 95th percentile values for motor and sensory UNSW were 0.5 ms in the proximal segment and 0.7 ms in the distal segment. CONCLUSIONS: When the 95-percentile value was considered as the normal upper limit, the criteria of abnormality for motor and sensory UNSW were greater than 0. 5 ms in the proximal segment and greater than 0.7 ms in the distal segment. SIGNIFICANCE: The normative values of UNSW may be useful in screening for ulnar neuropathy at the wrist. PMID- 16221564 TI - Spatial attention triggered by eye gaze enhances and speeds up visual processing in upper and lower visual fields beyond early striate visual processing. AB - OBJECTIVE: The detection of a lateralized visual target is faster when preceded by a face gazing to the location of this stimulus. Here we aimed to clarify the time-course of the visual processing modulated by these reflexive shifts of attention. METHODS: ERPs were measured on 16 subjects performing a speeded location task on a circular checkerboard. The checkerboard target appeared either on the left or right of the upper or lower visual field, and was preceded by a central face orienting its gaze obliquely to one of the four possible corner locations for the target to appear. RESULTS: Congruently cued targets were located faster than incongruently cued targets and were associated with larger and earlier occipital P1 (approximately 110 ms) and occipito-parieto-temporal N1 (approximately 150 ms) components. However, no such attentional modulations were found on the earlier C1 visual component, best observed with a negative polarity for upper visual field stimulations, and thought to originate largely from primary visual cortex. CONCLUSIONS AND SIGNIFICANCE: These results show that reflexive shifts of attention following oblique eye gaze to upper and lower visual fields increase and speed up the processing of visual information beyond the feedforward flow of information in primary visual cortex. PMID- 16221565 TI - The composite N1 component to gaps in noise. AB - OBJECTIVE: To indicate whether the double peaked N(1) to gaps in continuous white noise is a composite of onset and offset responses to transients or whether it reflects higher processing such as change or mismatch detection and to assess the role of attention in this process. METHODS: Evoked potentials were recorded to two binaural stimulus types: (1) gaps of different durations randomly distributed in continuous white noise; and (2) click pairs at intervals identical to those between gap onsets and offsets in the continuous noise stimulus. Potentials to these stimuli were recorded while subjects read a text and while detecting gaps in noise or click pairs. RESULTS: Potentials were detected to all click pairs and to gaps of 5 ms or longer, corresponding to the subjects' psychoacoustic gap detection threshold. With long gap durations of 200-800 ms, distinct potentials to gap onset and gap offset were observed. The waveforms to all click pairs and to offsets of long gaps were similar and single-peaked, while potentials to gaps of 10 ms and longer, and potentials to onsets of long gaps were double-peaked, consisting of two N(1) negativities, 60 ms apart, irrespective of gap duration. The first (N(1a)), was more frontal in its distribution and similar to that of clicks. The second (N(1b)) peak's distribution was more central/temporal and its source locations and time course of activity were distinct. No effects of attention on any of the varieties and constituents of N(1) were observed. CONCLUSIONS: Comparing potentials to gap onsets, to click pairs and to gap offsets, suggests that potentials to gap onsets involve not only sound onset/offset responses (N(1), N(1a)) but also the subsequent pre-attentive perception of the cessation of an ongoing sound (N(1b)). We propose that N(1b) is distinct from change or mismatch detection and is associated with termination of an ongoing continuous stimulus. We propose to call it the N(egation)-process. SIGNIFICANCE: A constituent of the N(1) complex is shown to be associated with the pre-attentive perception of termination of an ongoing stimulus and to have distinct scalp distribution and intracranial sources. PMID- 16221566 TI - Aging of phrenic nerve conduction in the elderly. AB - OBJECTIVE: We elucidated the possible relationship between age and conduction parameters of phrenic nerve in subjects above the sixth decade, comparing with the data from middle-age controls. METHODS: Diaphragmatic action potentials (DAPs) were recorded on bilateral hemithoraces of 41 volunteers aged 60-101 years (old group) and 25 volunteers aged 35-55 years (middle-age group). Statistical analyses were performed to assess the effects of aging on latency, latency corrected by size (Lat/Dist), amplitude, and the right-left difference of these DAP parameters. RESULTS: In all 61 subjects, age showed a significant quadratic correlation with latency and with Lat/Dist, and a linear correlation with amplitude. The right-left differences ranged from 0.0 to 14.5% for latency and from 6.5 to 112.4% for amplitude in the elderly. CONCLUSIONS: The normal ranges of DAP parameters should be determined according to age. The left-right difference may be a useful reference in diagnosing unilateral phrenic nerve lesion. SIGNIFICANCE: The precise normal ranges of phrenic nerve conduction parameters presented will encourage investigations of neuropathies in subjects aged above 60. PMID- 16221567 TI - NREM sleep alterations in narcolepsy/cataplexy. AB - OBJECTIVE: NREM sleep patterns of narcoleptic patients with cataplexy were studied, focusing on their sleep 'microstructure', by analyzing the cyclic alternating pattern (CAP). METHODS: Forty-nine HLA DQB1*0602-positive patients with narcolepsy/cataplexy (32 men and 17 women, aged 18-46 years) were included together with 37 age-matched normal controls. Each subject underwent one polysomnographic night recording after an adaptation night. Sleep stages were scored following standard criteria and CAP A phases were detected and classified into 3 subtypes (A1, A2, and A3). Power spectra for frequencies between 0.5 and 25 Hz were obtained for each CAP condition, separately in sleep stage 2 and SWS. RESULTS: Narcoleptic patients displayed reduced total CAP rate. A selective reduction in the number of A1 subtypes/hour and a reduced A3 index were found in narcoleptics who had also a smaller average number of CAP sequences. Narcoleptic patients had higher power spectra for fast frequencies mostly during SWS, while REM sleep power spectra showed significantly higher power density for frequency bins 0.5-1.5, 8.5-9.5, and 17.5-25 Hz. Similarly, CAP A1 subtypes and NCAP epochs during SWS displayed significantly higher power density for fast frequency bins. CONCLUSIONS: The main finding of this study is that the occurrence of the A1 CAP subtypes is impaired during NREM sleep in narcoleptic patients. Thus, narcolepsy seems to be accompanied not only by alterations of REM but also NREM sleep which is subtly but significantly impaired, as reflected by CAP and the corresponding EEG spectral analysis. SIGNIFICANCE: Our findings might indicate that in narcolepsy very-slow oscillation processes less effective than normal might be present, with a subtly impaired capability of grouping the other sleep EEG activities; this aspect deserves further insight in order to obtain a better understanding of its functional meaning. PMID- 16221568 TI - Caffeine effects on resting-state arousal. AB - OBJECTIVE: This study examined the use of caffeine to manipulate arousal level without the confounds associated with task-related activation. From previous work in our laboratory, an increase in skin conductance level (SCL) and EEG alpha frequency, together with a global decrease in alpha power, were used as markers of arousal increase, and we sought to identify these effects with caffeine ingestion. METHODS: We examined the effect of a single oral dose of caffeine (250 mg) in a randomised double-blind placebo-controlled repeated-measures cross-over study. Eighteen healthy university students (mean age 21 years; 13/18 females) participated in two sessions 1 week apart. EEG and autonomic data (SCL, heart rate, systolic and diastolic blood pressure, and respiration rate) from a 2 min eyes-closed epoch, commencing approximately 30 min after ingestion of caffeine or placebo, were examined. RESULTS: Caffeine was associated with increased SCL, a global reduction in EEG power in the alpha band, and a global increase in alpha frequency. There were no cardiovascular effects. CONCLUSIONS: The positive results are consistent with recent electrodermal and EEG studies of arousal and suggest that caffeine may be utilised as a task-free means of manipulating arousal in future investigations. Further work is necessary to clarify the absence of cardiovascular effects, and to integrate those data with emerging conceptualisations of arousal and activation. SIGNIFICANCE: The present data support the use of caffeine as a simple tool to explore the role of arousal in both normal and atypical functioning, and this may be useful in determining the validity and importance of supposed hyper- or hypo-arousal in such syndromes as attention-deficit/hyperactivity disorder (AD/HD). PMID- 16221569 TI - Auditory event related potentials and source current density estimation in phonologic/auditory dyslexics. AB - OBJECTIVE: To determine the generality of auditory processing impairment in phonologic dyslexics by studying their auditory Event-Related Potentials (ERPs) and the spatio-temporal distribution of their brain activity to auditory linguistic and non-linguistic stimuli with temporal and spectral discriminating cues. METHODS: Fourteen adult phonologic dyslexics and 14 normal reading students, all with high academic achievements, were compared. ERP waveform analysis and current density source estimation (Low resolution Electromagnetic Tomographic Analysis-LORETA) were conducted on 21-channel records from subjects who passively listened or actively discriminated 4 types of auditory stimuli: linguistic and non-linguistic stimuli that differed in spectral or temporal characteristics. RESULTS: Significant differences were found for all ERP latencies (N1, P2, N2, P3) in response to all stimuli, with dyslexics presenting longer latencies compared to normal readers. Current density distributions and their time courses also differed significantly, regardless of stimulus type or attention allocation. Among normal readers, early activity (around N1) was characterized by a rapid change of maximum activity from right to left temporal lobe. Later activity (around P3) was characterized by a stable temporal activity with bilaterally synchronous peak activity. Among the dyslexics, the early N1 activity was stable with left hemisphere prominence, with no alternation between the hemispheres, while the later P3 activity peaked earlier in the right hemisphere than in the left. CONCLUSIONS: Dyslexics were different from controls in processing all auditory stimuli: verbal and non-verbal stimuli with temporal as well as with spectral discriminating cues. The differences mainly consisted of latency and time courses of current density distributions, beginning as early as N1 and extending to the late P3. SIGNIFICANCE: Differences in processing auditory stimuli by phonologic dyslexics are not restricted to linguistic (phonological) stimuli, supporting a general auditory processing impairment in phonologic dyslexia. PMID- 16221571 TI - Short and middle-latency Median Nerve (MN) SEPs recorded by depth electrodes in human pre-SMA and SMA-proper. AB - OBJECTIVE: To analyse waveforms, latencies and amplitudes of Median Nerve (MN) SEPs recorded by stereotactically electrodes implanted in the SMA of 14 epileptic patients (9 in pre-SMA,3 in SMA-proper, 2 in both) in order to evaluate which short and middle-latency SEPs are generated in this area and which could be the physiological relevance of these responses. METHODS: Short and middle-latency MN SEPs were recorded by chronically implanted electrodes in the fronto-temporal cortex and in particular in the mesial frontal region of 14 drug-resistant epileptic patients. MN stimulations of 100 micros were delivered by skin electrodes at the wrist; stimulus intensity was adjusted slightly above the motor threshold. RESULTS: The main result of this study is that middle-latency SEPs were originated in pre-SMA but not in SMA-proper as demonstrated by both referential and bipolar recordings. In particular off-line computed bipolar traces between neighbouring contacts implanted in the pre-SMA and in the frontal external regions showed a phase reversal at the deepest contacts located in pre SMA. Conversely, bipolar recordings between neighbouring contacts implanted in the SMA-proper and in the frontal external regions showed inversion recovery at more superficial contacts, implanted in area 6. Finally, we confirmed that no short-latency MN SEP (and in particular the N30) is originated in the whole SMA. CONCLUSIONS: Among premotor areas, somatosensory inputs seem to reach pre-SMA and area 6 but not SMA-proper. SIGNIFICANCE: This study assessed that no scalp SEP in the first 100 ms after MN stimulus could be generated in SMA-proper. PMID- 16221570 TI - Gender difference in masseteric exteroceptive suppression period and pain perception. AB - OBJECTIVE: Use of brain stem reflexes in the assessment of orofacial function requires insight into the influence of demographic factors such as gender. The aim of this study was to characterize possible gender differences in the relation between quantitative measures of the masseteric exteroceptive suppression (ES) reflex response and pain perception evoked by incrementally increasing electrical stimulation. METHODS: In 12 men and 12 women, the surface electromyogram was recorded from the left masseter muscle. Thirteen fixed stimulus intensities from 5 to 35 mA at 2.5 mA increments were applied to the skin above the left mental nerve. The stimulation intensity at which the late ES appeared first and the first intensity at which the subjects reported the stimulus intensity to be painful were defined as the reflex threshold (RT) and pain threshold (PT), respectively. Furthermore, data were analyzed using stimulus-response curves, and the reflex appearance levels (RAL), the saturation level, the slope from appearance to saturation of the reflex (SLP), and the pain appearance level (PAL) were determined. RESULTS: The PT was equal to or higher than the RT in 9 of the 12 men, but only in 4 of the 12 women. Further, women had significantly lower PAL, RAL, and SLP (12.7 +/- 0.8, 12.9 +/- 1.4, and 3.0 +/- 0.9 mA, respectively) compared to men (20.3 +/- 1.6 mA, 16.7 +/- 1.1 T, and 4.1 +/- 0.4, respectively) (P < 0.05). CONCLUSIONS: The present results document that women have a lower reflex threshold and pain threshold to cutaneous electrical stimulation than men. SIGNIFICANCE: These findings suggest that gender differences may exist in the sensory-motor integration of primary afferent input from the orofacial region and that these differences should be considered in the design of future reflex studies. PMID- 16221572 TI - Computational detection of microRNAs targeting transcription factor genes in Arabidopsis thaliana. AB - MicroRNAs, an abundant class of tiny non-coding RNAs, have emerged as negative regulators for translational repression or cleavage of target mRNAs by the manner of complementary base paring in plants and animals. Recent studies have demonstrated that many known microRNAs have a remarkable propensity to target genes involved in development, particularly those of transcription factor genes. Therefore, an overall detection of Arabidopsis thaliana microRNAs targeting transcription factor genes will enhance greatly our understanding of microRNA biological functions in plant development. By searching short complementary sequences between transcription factor open-reading frames and intergenic region sequences, and considering RNA secondary structures and the sequence conversation between the genomes of Arabidopsis and Oryza sativa, we detected 96 candidate Arabidopsis microRNAs. These candidate microRNAs were predicted to target 102 transcription factor genes that are classified as 28 transcription factor gene families, particularly those of DNA-binding transcription factor families, which imply that microRNAs might be involved in complex transcriptional regulatory networks for specifying individual cell types in plant development. PMID- 16221573 TI - Vaginal vault prolapse: choice of operation. AB - The surgeon who faces a patient with vaginal vault prolapse is dealing with a complex and intriguing challenge. Part of the complexity is due to the lack of standardization and routine application of tools to assess pre- and postoperative anatomical and functional outcomes. Patient satisfaction is a major endpoint for surgical success; thus all aspects of the prolapse pathology and the patient's lifestyle should be considered. The surgeon needs to be well versed and flexible in order to choose the most appropriate operative approach to achieve optimal results for an individual patient. In this chapter we present the vaginal and abdominal approaches for the correction of vaginal vault prolapse, with discussion of the surgical outcomes and complications for each technique. A comprehensive comparison of the various techniques is offered on the basis of current published literature. In addition, we focus on various controversies, including the prevention of vault prolapse at the time of hysterectomy, issues regarding uterine preservation, the management of overt or occult concomitant stress incontinence, and the place-if any-for combined anti-incontinence procedures at the time of prolapse surgery. New minimally invasive techniques for vault prolapse are also reviewed. We emphasize areas that call for further research and for standardized outcome criteria. PMID- 16221574 TI - Quantification of betamethasone in human plasma by liquid chromatography-tandem mass spectrometry using atmospheric pressure photoionization in negative mode. AB - Betamethasone is a synthetic corticosteroid designed to exert a marked glucocorticoid activity. As the free alcohol, betamethasone finds widespread clinical applications related to its anti-inflammatory and immunosuppressant activity. In the present study, a fast, sensitive, robust method was developed for the determination and quantification of betamethasone in human plasma by liquid chromatography coupled with tandem mass spectrometry, using photospray ionization in negative mode. Betamethasone was extracted from 0.5 ml human plasma by liquid-liquid extraction (LLE) using chloramphenicol as internal standard. The method has a chromatographic run of 2.5 min using a C(18) analytical column (100 mm x 2.1 mm i.d.) and the linear calibration curve over the range was linear from 0.05 to 50 ng ml(-1) (r(2)>0.993). The between-run precision, based on the relative standard deviation replicate quality controls was 94.1% (0.15 ng ml( 1)), 90.7% (4.0 ng ml(-1)) and 97.2% (40 ng ml(-1)). The between-run accuracy for the above-mentioned concentrations was 11.9, 9.0 and 9.8%, respectively. The method herein described was employed in a bioequivalence study of two formulations of dexchlorpheniramine/betamethasone 2 mg/0.25 mg tablets. PMID- 16221575 TI - Development of a solid phase microextraction-gas chromatography method to determine N-hydroxymethyl-N-methylformamide and N-methylformamide in urine. AB - A headspace solid phase microextraction (SPME) method has been developed to determine metabolites of dimethylformamide, N-hydroxymethyl-N-methylformamide, and N-methylformamide (NMF) as NMF in urine by gas chromatography with nitrogen phosphorus detector (GC-NPD). An SPME holder with a 65-microm PDMS/DVB fiber coating was used. Optimal desorption conditions were 250 degrees C for 1 min, adsorption at 80 degrees C for 15 min, and 3.00 mL of sample in the headspace vial. The method presented good resolution, repeatability, recovery, detection limit, ruggedness and response linearity. PMID- 16221576 TI - Validity and reliability of the Italian version of the Chronic Liver Disease Questionnaire (CLDQ-I) for the assessment of health-related quality of life. AB - BACKGROUND: The Chronic Liver Disease Questionnaire is a specific health-related quality of life assessment designed for patients with liver diseases. AIM: The aim of this paper is to report on the validity, reliability and sensitivity to change of the Italian version (Chronic Liver Disease Questionnaire-I) in subjects with HCV infection. SUBJECTS: The Chronic Liver Disease Questionnaire-I was administered to 350 subjects with HCV infection together with the World Health Organization Quality of Life Assessment, abbreviated version, a generic quality of life assessment. METHODS: The instrument was translated from English, backtranslated and reviewed in focus groups in the framework of a large multicentre study. Exploratory factor analysis identified five factors accounting for 65% of the variance of Chronic Liver Disease Questionnaire-I items and only partially overlapping with those found in the original version. RESULTS: The Chronic Liver Disease Questionnaire-I proved to discriminate between subjects with and without comorbid diseases at baseline (t-test = 3.59, p < 0.001). Test retest reliability was moderate (ICC = 0.60). The Chronic Liver Disease Questionnaire-I was sensitive to change in patients who deteriorated after one month of treatment. Change in the overall Chronic Liver Disease Questionnaire-I score in deteriorated patients was correlated with changes in World Health Organization Quality of Life Assessment, abbreviated version scores in the physical, psychological and environment, but not in the social area. CONCLUSIONS: The Italian version of Chronic Liver Disease Questionnaire is a valid and reliable instrument to be used in cross-sectional and longitudinal studies. PMID- 16221577 TI - Metabolic genomics. PMID- 16221578 TI - How Escherichia coli and Saccharomyces cerevisiae build Fe/S proteins. AB - Owing to the versatile electronic properties of iron and sulfur, iron sulfur (Fe/S) clusters are perfectly suited for sensing changes in environmental conditions and regulating protein properties accordingly. Fe/S proteins have been recruited in a wide array of diverse biological processes, including electron transfer chains, metabolic pathways and gene regulatory circuits. Chemistry has revealed the great diversity of Fe/S clusters occurring in proteins. The question now is to understand how iron and sulfur come together to form Fe/S clusters and how these clusters are subsequently inserted into apoproteins. Iron, sulfide and reducing conditions were found to be sufficient for successful maturation of many apoproteins in vitro, opening the possibility that insertion might be a spontaneous event. However, as in many other biological pathways such as protein folding, genetic analyses revealed that Fe/S cluster biogenesis and insertion depend in vivo upon auxiliary proteins. This was brought to light by studies on Azotobacter vinelandii nitrogenase, which, in particular, led to the concept of scaffold proteins, the role of which would be to allow transient assembly of Fe/S cluster. These studies paved the way toward the identification of the ISC and SUF systems, subjects of the present review that allow Fe/S cluster assembly into apoproteins of most organisms. Despite the recent discovery of the SUF and ISC systems, remarkable progress has been made in our understanding of their molecular composition and biochemical mechanisms. Such a rapid increase in our knowledge arose from a convergent interest from researchers engaged in unrelated fields and whose complementary expertise covered most experimental approaches used in biology. Also, the high conservation of ISC and SUF systems throughout a wide array of organisms helped cross-feeding between studies. The ISC system is conserved in eubacteria and most eukaryotes, while the SUF system arises in eubacteria, archaea, plants and parasites. ISC and SUF systems share a common core function made of a cysteine desulfurase, which acts as a sulfur donor, and scaffold proteins, which act as sulfur and iron acceptors. The ISC and SUF systems also exhibit important differences. In particular, the ISC system includes an Hsp70/Hsp40-like pair of chaperones, while the SUF system involves an unorthodox ATP-binding cassette (ABC)-like component. The role of these two sets of ATP-hydrolyzing proteins in Fe/S cluster biogenesis remains unclear. Both systems are likely to target overlapping sets of apoproteins. However, regulation and phenotypic studies in E. coli, which synthesizes both types of systems, leads us to envisage ISC as the house-keeping one that functions under normal laboratory conditions, while the SUF system appears to be required in harsh environmental conditions such as oxidative stress and iron starvation. In Saccharomyces cerevisiae, the ISC system is located in the mitochondria and its function is necessary for maturation of both mitochondrial and cytosolic Fe/S proteins. Here, we attempt to provide the first comprehensive review of the ISC and SUF systems since their discovery in the mid and late 1990s. Most emphasis is put on E. coli and S. cerevisiae models with reference to other organisms when their analysis provided us with information of particular significance. We aim at covering information made available on each Isc and Suf component by the different experimental approaches, including physiology, gene regulation, genetics, enzymology, biophysics and structural biology. It is our hope that this parallel coverage will facilitate the identification of both similarities and specificities of ISC and SUF systems. PMID- 16221579 TI - Function, attachment and synthesis of lipoic acid in Escherichia coli. AB - A series of genetic, biochemical, and physiological studies in Escherichia coli have elucidated the unusual pathway whereby lipoic acid is synthesized. Here we describe the results of these investigations as well as the functions of enzyme proteins that are modified by covalent attachment of lipoic acid and the enzymes that catalyze the modification reactions. Some aspects of the synthesis and attachment mechanisms have strong parallels in the pathways used in synthesis and attachment of biotin and these are compared and contrasted. Homologues of the lipoic acid metabolism proteins are found in all branches of life, save the Archea, and thus these findings seem to have wide biological relevance. PMID- 16221580 TI - Microbial dimethylsulfoxide and trimethylamine-N-oxide respiration. AB - Over the last two decades, the biochemistry and genetics of dimethylsulfoxide (DMSO) and trimethylamine-N-oxide (TMAO) respiration has been characterised, particularly in Escherichia coli marine bacteria of the genus Shewanella and the purple phototrophic bacteria, Rhodobacter sphaeroides and R. capsulatus. All of the enzymes (or catalytic subunits) involved the final step in DMSO and TMAO respiration contain a pterin molybdenum cofactor and are members of the DMSO reductase family of molybdoenzymes. In E. coli, the dimethylsulfoxide reductase (DmsABC) can be purified from membranes as a complex, which exhibits quinol-DMSO oxidoreductase activity. The enzyme is anchored to the membrane via the DmsC subunit and its catalytic subunit DmsA is now considered to face the periplasm. Electron transfer to DmsA involves the DmsB subunit, which is a polyferredoxin related to subunits found in other molybdoenzymes such as nitrate reductase and formate dehydrogenase. A characteristic of the DmsAB-type DMSO reductase is its ability to reduce a variety of S- and N-oxides. E. coli contains a trimethylamine N-oxide reductase (TorA) that is highly specific for N-oxides. This enzyme is located in the periplasm and is connected to the quinone pool via a membrane bound penta-haem cytochrome (TorC). DorCA in purple phototrophic bacteria of the genus Rhodobacter is very similar to TorCA with the critical difference that DorA catalyses reduction of both DMSO and TMAO. It is known as a DMSO reductase because the S-oxide is the best substrate. Crystal structures of DorA and TorA have revealed critical differences at the Mo active site that may explain the differences between substrate specificity between the two enzymes. DmsA, TorA and DorA possess a "twin arginine" N-terminal signal sequence consistent with their secretion via the TAT secretory system and not the Sec system. The enzymes are secreted with their bound prosthetic groups: this take place in the cytoplasm and the biogenesis involves a chaperone protein, which is cognate for each enzyme. Expression of the DMSO and TMAO respiratory operons is induced in response to a fall in oxygen tension. dmsABC expression is positively controlled by the oxygen responsive transcription factor, Fnr and ModE, a transcription factor that binds molybdate. In contrast, torCAD expression is not under Fnr- or ModE-control but is dependent upon a sensor histidine kinase-response regulator pair, TorSR, which activate gene expression under conditions of low oxygen tension in the presence of N- or S-oxide. Regulation of dorCDA expression is similar to that seen for torCAD but it appears that the expression of the sensor histidine kinase-response regulator pair, DorSR is regulated by Fnr and there is an additional tier of regulation involving the ModE-homologue MopB, molybdate and the transcription factor DorX. Analysis of microbial genomes has revealed the presence of dms and tor operons in a wide variety of bacteria and in some archaea and duplicate dms and tor operons have been identified in E. coli. Challenges ahead will include the determination of the significance of the presence of the dms operon in bacterial pathogens and the determination of the significance of DMSO respiration in the global turnover of marine organo-sulfur compounds. PMID- 16221581 TI - Energy metabolism and its compartmentation in Trypanosoma brucei. AB - African trypanosomes are parasitic protozoa of the order of Kinetoplastida, which cause sleeping sickness and nagana. Trypanosomes are not only of scientific interest because of their clinical importance, but also because these protozoa contain several very unusual biological features, such as their special energy metabolism. The energy metabolism of Trypanosoma brucei differs significantly from that of its host, not only because it comprises distinct enzymes and metabolic pathways, but also because some of the glycolytic enzymes are localized in organelles called glycosomes. Furthermore, the energy metabolism changes drastically during the complex life cycle of this parasite. This review will focus on the recent advances made in understanding the process of ATP production in T. brucei during its life cycle and the consequences of the special subcellular compartmentation. PMID- 16221582 TI - The first cell. AB - The First Cell arose in the previously pre-biotic world with the coming together of several entities that gave a single vesicle the unique chance to carry out three essential and quite different life processes. These were: (a) to copy informational macromolecules, (b) to carry out specific catalytic functions, and (c) to couple energy from the environment into usable chemical forms. These would foster subsequent cellular evolution and metabolism. Each of these three essential processes probably originated and was lost many times prior to The First Cell, but only when these three occurred together was life jump-started and Darwinian evolution of organisms began. The replication of informational molecules that made only occasional mistakes allowed evolution to form all the basic components of cellular life. Ribozymes, the first informational molecules, were also catalytic. Energy coupling required the formation of a closed lipid surface to generate and maintain an ion-motive gradient. The closed vesicle partitioned components and avoided dilution within the primordial sea. Closed membranes were essential for the first self-reproducing cell to arise and for its descendants to disperse. Subsequent cellular development after the origin of The First Cell led to the beginnings of intermediary metabolism and membrane transport processes. This long process, subject to strong evolutionary selection, developed the cellular biology that is now shared by all extant organisms. PMID- 16221583 TI - Neuroanatomy of the oculomotor system. Preface. PMID- 16221584 TI - Present concepts of oculomotor organization. AB - This chapter gives an introduction to the oculomotor system, thus providing a framework for the subsequent chapters. This chapter describes the characteristics, and outlines the structures involved, of the five basic types of eye movements, for gaze holding ("neural integrator") and eye movements in three dimensions (Listing's law, pulleys). PMID- 16221585 TI - Biological organization of the extraocular muscles. AB - Extraocular muscle is fundamentally distinct from other skeletal muscles. Here, we review the biological organization of the extraocular muscles with the intent of understanding this novel muscle group in the context of oculomotor system function. The specific objectives of this review are threefold. The first objective is to understand the anatomic arrangement of the extraocular muscles and their compartmental or layered organization in the context of a new concept of orbital mechanics, the active pulley hypothesis. The second objective is to present an integrated view of the morphologic, cellular, and molecular differences between extraocular and the more traditional skeletal muscles. The third objective is to relate recent data from functional and molecular biology studies to the established extraocular muscle fiber types. Developmental mechanisms that may be responsible for the divergence of the eye muscles from a skeletal muscle prototype also are considered. Taken together, a multidisciplinary understanding of extraocular muscle biology in health and disease provides insights into oculomotor system function and malfunction. Moreover, because the eye muscles are selectively involved or spared in a variety of neuromuscular diseases, knowledge of their biology may improve current pathogenic models of and treatments for devastating systemic diseases. PMID- 16221586 TI - Sensory control of extraocular muscles. AB - The role of sensory receptors in eye muscles is not well understood, but there is physiological and clinical evidence for the presence of proprioceptive signals in many areas of the central nervous system. It is unclear which structures generate these sensory signals, and which central neural pathways are involved. Three different types of receptors are associated with eye muscles: (1) muscle spindles, (2) palisade endings, and (3) Golgi tendon organs, but their occurrence varies wildly between species. A review of their organization shows that each receptor is mainly confined to a morphologically separate layer of the eye muscle. The palisade endings - which are unique to eye muscles, are associated with the global layer; and they have been found in all mammals studied so far. Their function is unknown. The muscle spindles, if they are present in a species, lie in the orbital layer, or at its junction to the global layer. Golgi tendon organs appear to be unique to artiodactyls (i.e., sheep and goats, etc.); they lie in an outer distal marginal layer of the eye muscle, called the "peripheral patch layer" in sheep. The specific association between palisade endings and the multiply innervated type of muscle fibers of the global layer has led to the hypothesis that together they may act as a sensory receptor, and provide a source of central proprioceptive signals. But other interpretations of the morphological evidence do not support this role. PMID- 16221587 TI - The extraocular motor nuclei: organization and functional neuroanatomy. AB - The organization of the motoneuron subgroups in the brainstem controlling each extraocular eye muscle is highly stable through the vertebrate species. The subgroups are topographically organized in the oculomotor nucleus (III) and are usually considered to form the final common pathway for eye muscle control. Eye muscles contain a unique type of slow non-twitch, fatigue-resistant muscle fiber, the multiply innervated muscle fibers (MIFs). The recent identification the MIF motoneurons shows that they too have topographic organization, but very different from the classical singly innervated muscle fiber (SIF) motoneurons. The MIF motoneurons lie around the periphery of the oculomotor nucleus (III), trochlear nucleus (IV), and abducens nucleus (VI), slightly separated from the SIF subgroups. The location of four different types of neurons in VI are described and illustrated: (1) SIF motoneurons, (2) MIF motoneurons, (3) internuclear neurons, and (4) the paramedian tract neurons which project to the flocculus. Afferents to the motoneurons arise from the vestibular nuclei, the oculomotor and abducens internuclear neurons, the mesencephalic and pontine burst neurons, the interstitial nucleus of Cajal, nucleus prepositus hypoglossi, the supraoculomotor area and the central mesencephalic reticular formation and the pretectum. The MIF and SIF motoneurons have different histochemical properties and different afferent inputs. The hypothesis that SIFs participate in moving the eye and MIFs determine the alignment seems possible but is not compatible with the concept of a final common pathway. PMID- 16221588 TI - The reticular formation. AB - The reticular formation of the brainstem contains functional cell groups that are important for the control of eye, head, or lid movements. The mesencephalic reticular formation is primarily involved in the control of vertical gaze, the paramedian pontine reticular formation in horizontal gaze, and the medullary pontine reticular formation in head movements and gaze holding. In this chapter, the locations, connections, and histochemical properties of the functional cell groups are reviewed and correlated with specific subdivisions of the reticular formation. PMID- 16221589 TI - The anatomy of the vestibular nuclei. AB - The vestibular portion of the eighth cranial nerve informs the brain about the linear and angular movements of the head in space and the position of the head with respect to gravity. The termination sites of these eighth nerve afferents define the territory of the vestibular nuclei in the brainstem. (There is also a subset of afferents that project directly to the cerebellum.) This chapter reviews the anatomical organization of the vestibular nuclei, and the anatomy of the pathways from the nuclei to various target areas in the brain. The cytoarchitectonics of the vestibular brainstem are discussed, since these features have been used to distinguish the individual nuclei. The neurochemical phenotype of vestibular neurons and pathways are also summarized because the chemical anatomy of the system contributes to its signal-processing capabilities. Similarly, the morphologic features of short-axon local circuit neurons and long axon cells with extrinsic projections are described in detail, since these structural attributes of the neurons are critical to their functional potential. Finally, the composition and hodology of the afferent and efferent pathways of the vestibular nuclei are discussed. In sum, this chapter reviews the morphology, chemoanatomy, connectivity, and synaptology of the vestibular nuclei. PMID- 16221590 TI - Nucleus prepositus. AB - The cytoarchitecture and the histochemistry of nucleus prepositus hypoglossi and its afferent and efferent connections to oculomotor structures are described. The functional significance of the afferent connections of the nucleus is discussed in terms of current knowledge of the firing behavior of prepositus neurons in alert animals. The efferent connections of the nucleus and the results of lesion experiments suggest that it plays a role in a variety of functions related to the control of gaze. PMID- 16221591 TI - Oculomotor cerebellum. AB - The anatomical, physiological, and behavioral evidence for the involvement of three regions of the cerebellum in oculomotor behavior is reviewed here: (1) the oculomotor vermis and paravermis of lobules V, IV, and VII; (2) the uvula and nodulus; (3) flocculus and ventral paraflocculus. No region of the cerebellum controls eye movements exclusively, but each receives sensory information relevant for the control of multiple systems. An analysis of the microcircuitry suggests how sagittal climbing fiber zones bring visual information to the oculomotor vermis; convey vestibular information to the uvula and nodulus, while optokinetic space is represented in the flocculus. The mossy fiber projections are more heterogeneous. The importance of the inferior olive in modulating Purkinje cell responses is discussed. PMID- 16221592 TI - Inferior olive and oculomotor system. AB - Three subnuclei within the inferior olive are implicated in the control of eye movement; the dorsal cap (DC), the beta-nucleus and the dorsomedial cell column (DMCC). Each of these subnuclei can be further divided into clusters of cells that encode specific parameters of optokinetic and vestibular stimulation. DC neurons respond to optokinetic stimulation in one of three planes, corresponding to the anatomical planes of the semicircular canals. Neurons in the beta-nucleus and DMCC respond to vestibular stimulation in the planes of the vertical semicircular canals and otoliths. Each these olivary nuclei receives excitatory and inhibitory signals from pre-olivary structures. The DC receives excitatory signals from the ipsilateral nucleus of the optic tract (NOT) and inhibitory signals from the contralateral nucleus prepositus hypoglossi (NPH). The beta nucleus and DMCC receive inhibitory signals from the ipsilateral nucleus parasolitarius (Psol) and excitatory signals from the contralateral dorsal Y group. Consequently, the olivary projection to the cerebellum, although totally crossed, still represents bilateral sensory stimulation. Inputs to the inferior olive from the NOT, NPH, Psol or Y-group discharge at frequencies of 10-100 imp/s. CFRs discharge at 1-5 imp/s; a frequency reduction of an order of magnitude. Inferior olivary projections to the contralateral cerebellum are sagittally arrayed onto multiple cerebellar folia. These arrays establish coordinate systems in the flocculus and nodulus, representing head-body movement. These climbing fiber-defined spatial coordinate systems align Purkinje cell discharge onto subjacent cerebellar and vestibular nuclei. In the oculomotor system, olivo-cerebellar circuitry enhances and modifies eye movements based on movement of the head-body in space. PMID- 16221593 TI - The oculomotor role of the pontine nuclei and the nucleus reticularis tegmenti pontis. AB - Cerebral cortex and the cerebellum interact closely in order to facilitate spatial orientation and the generation of motor behavior, including eye movements. This interaction is based on a massive projection system that allows the exchange of signals between the two cortices. This cerebro-cerebellar communication system includes several intercalated brain stem nuclei, whose eminent role in the organization of oculomotor behavior has only recently become apparent. This review focuses on the two major nuclei of this group taking a precerebellar position, the pontine nuclei and the nucleus reticularis tegmenti pontis, both intimately involved in the visual guidance of eye movements. PMID- 16221594 TI - The mammalian superior colliculus: laminar structure and connections. AB - The superior colliculus is a laminated midbrain structure that acts as one of the centers organizing gaze movements. This review will concentrate on sensory and motor inputs to the superior colliculus, on its internal circuitry, and on its connections with other brainstem gaze centers, as well as its extensive outputs to those structures with which it is reciprocally connected. This will be done in the context of its laminar arrangement. Specifically, the superficial layers receive direct retinal input, and are primarily visual sensory in nature. They project upon the visual thalamus and pretectum to influence visual perception. These visual layers also project upon the deeper layers, which are both multimodal, and premotor in nature. Thus, the deep layers receive input from both somatosensory and auditory sources, as well as from the basal ganglia and cerebellum. Sensory, association, and motor areas of cerebral cortex provide another major source of collicular input, particularly in more encephalized species. For example, visual sensory cortex terminates superficially, while the eye fields target the deeper layers. The deeper layers are themselves the source of a major projection by way of the predorsal bundle which contributes collicular target information to the brainstem structures containing gaze-related burst neurons, and the spinal cord and medullary reticular formation regions that produce head turning. PMID- 16221595 TI - The pretectum: connections and oculomotor-related roles. AB - Research over the past two decades in mammals, especially primates, has greatly improved our understanding of the afferent and efferent connections of two retinorecipient pretectal nuclei, the nucleus of the optic tract (NOT) and the pretectal olivary nucleus (PON). Functional studies of these two nuclei have further elucidated some of the roles that they play both in oculomotor control and in relaying oculomotor-related signals to visual relay nuclei. Therefore, following a brief overview of the anatomy and retinal projections to the entire mammalian pretectum, the connections and potential roles of the NOT and the PON are considered in detail. Data on the specific connections of the NOT are combined with data from single-unit recording, microstimulation, and lesion studies to show that this nucleus plays critical roles in optokinetic nystagmus, short-latency ocular following, smooth pursuit eye movements, and adaptation of the gain of the horizontal vestibulo-ocular reflex. Comparable data for the PON show that this nucleus plays critical roles in the pupillary light reflex, light evoked blinks, rapid eye movement sleep triggering, and modulating subcortical nuclei involved in circadian rhythms. PMID- 16221597 TI - Oculomotor-related pathways of the basal ganglia. PMID- 16221596 TI - The accessory optic system: basic organization with an update on connectivity, neurochemistry, and function. AB - The accessory optic system (AOS) is formed by a series of terminal nuclei receiving direct visual information from the retina via one or more accessory optic tracts. In addition to the retinal input, derived from ganglion cells that characteristically have large receptive fields, are direction-selective, and have a preference for slow moving stimuli, there are now well-characterized afferent connections with a key pretectal nucleus (nucleus of the optic tract) and the ventral lateral geniculate nucleus. The efferent connections of the AOS are robust, targeting brainstem and other structures in support of visual-oculomotor events such as optokinetic nystagmus and visual-vestibular interaction. This chapter reviews the newer experimental findings while including older data concerning the structural and functional organization of the AOS. We then consider the ontogeny and phylogeny of the AOS and include a discussion of similarities and differences in the anatomical organization of the AOS in nonmammalian and mammalian species. This is followed by sections dealing with retinal and cerebral cortical afferents to the AOS nuclei, interneuronal connections of AOS neurons, and the efferents of the AOS nuclei. We conclude with a section on Functional Considerations dealing with the issues of the response properties of AOS neurons, lesion and metabolic studies, and the AOS and spatial cognition. PMID- 16221598 TI - Cortico-cortical networks and cortico-subcortical loops for the higher control of eye movements. AB - There are multiple distinct regions, or eye fields, in the cerebral cortex that contribute directly to the initiation and control of voluntary eye movements. We concentrate on six of these: the frontal eye field, parietal eye field, supplementary eye field, middle superior temporal area, prefrontal eye field, and area 7 m (precuneus in humans). In each of these regions: (1) there is neural activity closely related to eye movements; (2) electrical microstimulation produces or modifies eye movements; (3) surgical lesions or chemical inactivation impairs eye movements; (4) there are direct neural projections to major structures in the brainstem oculomotor system; and (5) increased activity is observed during eye movement tasks in functional magnetic resonance imaging or positron emission tomography experiments in humans. Each of these eye fields is reciprocally connected with the other eye fields, and each receives visual information directly from visual association cortex. Each eye field has distinct subregions that are concerned with either saccadic or pursuit eye movements. The saccadic subregions are preferentially interconnected with other saccade subregions and the pursuit subregions are preferentially interconnected with other pursuit subregions. Current evidence strongly supports the proposal that there are parallel cortico-cortical networks that control purposeful saccadic and pursuit eye movements, and that the activity in those networks is modulated by feedback information, via the thalamus, from the superior colliculus, basal ganglia, and cerebellum. PMID- 16221599 TI - MRI and fMRI analysis of oculomotor function. AB - This chapter reviews the anatomical correlations of the cortical oculomotor centers in humans. The modern structural methods allow a better anatomical definition of the parietal, frontal and temporal structures involved in oculomotor control. Functional imaging reveals the cortical networks involved in saccadic, pursuit, and vestibular eye movements. Finally, the interaction of the network between attention and eye movements is discussed. PMID- 16221600 TI - Long descending motor tract axons and their control of neck and axial muscles. AB - It has been tacitly assumed that a long descending motor tract axon consists of a private line connecting the cell of origin to a single muscle, as a motoneuron innervates a single muscle. However, this notion of a long descending motor tract referred to as a private line is no longer tenable, since recent studies have showed that axons of all major long descending motor tracts send their axon collaterals to multiple spinal segments, suggesting that they may exert simultaneous influences on different groups of spinal interneurons and motoneurons of multiple muscles. The long descending motor systems are divided into two groups, the medial and the lateral systems including interneurons and motoneurons. In this chapter, we focus mainly on the medial system (vestibulospinal, reticulospinal and tectospinal systems) in relation to movement control of the neck, describe the intraspinal morphologies of single long descending motor tract axons that are stained with intracellular injection of horseradish peroxidase, and provide evidence that single long motor-tract neurons are implicated in the neural implementation of functional synergies for head movements. PMID- 16221605 TI - The European clinical trials directive-a regulatory approach for filing drug substance information. PMID- 16221604 TI - Deciphering amphibian diversity through DNA barcoding: chances and challenges. AB - Amphibians globally are in decline, yet there is still a tremendous amount of unrecognized diversity, calling for an acceleration of taxonomic exploration. This process will be greatly facilitated by a DNA barcoding system; however, the mitochondrial population structure of many amphibian species presents numerous challenges to such a standardized, single locus, approach. Here we analyse intra- and interspecific patterns of mitochondrial variation in two distantly related groups of amphibians, mantellid frogs and salamanders, to determine the promise of DNA barcoding with cytochrome oxidase subunit I (cox1) sequences in this taxon. High intraspecific cox1 divergences of 7-14% were observed (18% in one case) within the whole set of amphibian sequences analysed. These high values are not caused by particularly high substitution rates of this gene but by generally deep mitochondrial divergences within and among amphibian species. Despite these high divergences, cox1 sequences were able to correctly identify species including disparate geographic variants. The main problems with cox1 barcoding of amphibians are (i) the high variability of priming sites that hinder the application of universal primers to all species and (ii) the observed distinct overlap of intraspecific and interspecific divergence values, which implies difficulties in the definition of threshold values to identify candidate species. Common discordances between geographical signatures of mitochondrial and nuclear markers in amphibians indicate that a single-locus approach can be problematic when high accuracy of DNA barcoding is required. We suggest that a number of mitochondrial and nuclear genes may be used as DNA barcoding markers to complement cox1. PMID- 16221606 TI - Solid state and dissolution rate characterization of co-ground mixtures of nifedipine and hydrophilic carriers. AB - Co-ground powders of the poorly water-soluble drug nifedipine and a hydrophilic carrier, [partially hydrolyzed gelatin (PHG), polyvinylpyrrolidone (PVP), sodium dodecyl sulfate (SDS), hydroxypropyl methylcellulose (HPMC), polyethylene glycol (PEG), urea or Pluronic F108] were prepared in order to improve the dissolution rate of nifedipine. The effects of type of grinding equipment, grinding time, and type of hydrophilic carrier on the crystallinity of nifedipine (x-ray diffraction and differential scanning calorimetry) on the interaction between drug and carriers (differential scanning calorimetry), on the particle size and appearance (scanning electron microscopy), on the wettability (contact angle measurements), and on the drug release were investigated. Grinding nifedipine together with these carriers improved the dissolution rate. PHG-ground mixtures resulted in the fastest dissolution rate followed by PVP, SDS, HPMC, Pluronic, urea, and PEG. This effect was not only due to particle size reduction, which increased in the order PHGBO 2=IPM>LP>PEG400, and increased with LID solubility in the oily solvents, although LID crystals were dispersed in all solvents. Subsequently, oily formulations that consisted of different ratios of the first oily solvent (IPM, BO-2, or DES) (each 0-20%), the second oily solvent (LP) and an oily mixture of microcrystalline wax/white petrolatum/paraffin (1/5/4) were evaluated. BO-2 groups at a concentration of 5% and 10% had the highest Japp among the oily formulations, although a higher BO-2 resulted in lower skin permeation. In addition, pretreatment with BO-2 increased the skin permeation of LID. These results suggest that the penetration enhancing effect by the system may be related to the skin penetration of BO-2 itself. Finally, mathematical analysis was done to evaluate the effect of BO-2, and it was shown that BO-2 improved the LID solubility in stratum corneum lipids to efficiently enhance the LID permeation through skin. PMID- 16221608 TI - Performance of multilayered particles: influence of a thin cushioning layer. AB - Nowadays, oral dosage forms with controlled release kinetics have known an increasing interest. The polymer coating of drug-loaded particles is one of the most common methods used for controlling drug delivery. Such multilayered particles could be either filled into capsules or compressed into tablets for their oral administration. However, many studies have noticed that coating films are damaged during the compression process, leading to significant changes in drug release profiles. The aims of this study were to investigate the effects of a thin cushioning layer [made of HydroxyPropylMethyl Cellulose (HPMC)] applied on coated theophylline particles upon particle characteristics, tablet properties, and then upon their dissolution performance. If no significant effect was shown with particles, this thin HPMC layer played an important role in the tablets. Tablet cohesiveness was decreased due to HPMC cushioning properties and moreover, the theophylline release rate was increased, as HPMC is a water-soluble polymer creating channels in polymer film for dissolution medium. Therefore, a cushioning layer helped to protect polymer coats from fracture during compression but could also affect drug release and so, both effects must be checked in such a drug delivery system. PMID- 16221610 TI - Cell lines as in vitro models for drug screening and toxicity studies. AB - Cell culture is highly desirable, as it provides systems for ready, direct access and evaluation of tissues. The use of tissue culture is a valuable tool to study problems of clinical relevance, especially those related to diseases, screening, and studies of cell toxicity mechanisms. Ready access to the cells provides the possibility for easy studies of cellular mechanisms that may suggest new potential drug targets and, in the case of pathological-derived tissue, it has an interesting application in the evaluation of therapeutic agents that potentially may treat the dysfunction. However, special considerations must be addressed to establish stable in vitro function. In primary culture, these factors are primarily linked to greater demands of tissue to adequately survive and develop differentiated conditions in vitro. Additional requirements include the use of special substrates (collagen, laminin, extracellular matrix preparations, etc.), growth factors and soluble media supplements, some of which can be quite complex in their composition. These demands, along with difficulties in obtaining adequate tissue amounts, have prompted interest in developing immortalized cell lines which can provide unlimited tissue amounts. However, cell lines tend to exhibit problems in stability and/or viability, though they serve as a feasible alternative, especially regarding new potential applications in cell transplant therapy. In this regard, stem cells may also be a source for the generation of various cell types in vitro. This review will address aspects of cell culture system application, with focus on immortalized cell lines, in studying cell function and dysfunction with the primary aim being to identify cell targets for drug screening. PMID- 16221609 TI - The physicochemical characteristics of freeze-dried scutellarin-cyclodextrin tetracomponent complexes. AB - In an effort to improve the solubility of the insoluble drug scutellarin, a novel complexation of scutellarin with beta-cyclodextrin (beta-CD) was studied. Tetracomponent freeze-dried complex was prepared with scutellarin, beta-CD, Hydroxypropyl Methylcellulose (HPMC), and triethanolamine. To confirm complex formation, complex was characterized by Fourier transform infrared spectroscopy (FT-IR), powder X-ray diffraction, and differential scanning calorimetry (DSC). Phase-solubility analysis suggested the soluble complexes having 1:1 stoichiometry. The beta-CD solubilization of scutellarin could be improved significantly by combining water-soluble polymer and pH adjuster. Comparing the binary, ternary solid systems with tetrary systems, tetracomponent freeze-dried complex showed the best effect of solubilization. A maximal solubility of scutellarin (23.65 mg/ml) was achieved with tetracomponent freeze-dried complex, up to 148-fold increase over scutellarin solubility in water, and the solubility of scutellarin is 15.35 microg/ml (up to 6-fold) in simulated gastric fluid. PMID- 16221611 TI - Characterization and body distribution of beta-elemene solid lipid nanoparticles (SLN). AB - Solid lipid nanoparticles (SLN) containing beta-elemene, a volatile oil used for the treatment of cancer, were prepared by the method combining probe sonication and membrane extrusion. Effects of the formulations and procedures on the characteristics of SLN were investigated. Body distribution of beta-elemene SLN in rats after intravenous administration was compared with that of the commercial emulsion. The results showed that dispersing the surfactant in the melted lipid matrix could obtain smaller particles than that dispersing in the water phase. Increasing the ratio of monostearin in the lipid matrix or the concentration of surfactant reduced the mean volume size of the SLN. Optimized formulation was composed of monostearin and precirol ATO 5 at a mass ratio of 3:7, which was quite stable for 8 months at room temperature. In vitro release of beta-elemene from the SLN was slow and stable without obvious burst release and was found to follow the Higuich equation. After intravenous administration, the beta-elemene levels after 5 min injection of SLN formulation were 1.5, 2.9, and 1.4 times higher than those of beta-elemene emulsion in liver, spleen, and kidney, respectively, while the concentrations of beta-elemene were decreased 30% in heart and lung. Therefore, the SLN containing beta-elemene might be an attractive candidate for the treatment of liver cancer. PMID- 16221612 TI - A new stability--indicating RP-HPLC method to determine assay and known impurity of Celecoxib API. AB - A simple, rapid and accurate Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed to determine assay and known impurity of Celecoxib API. The chromatographic separation was performed on reversed-phase C-18 column. Eluents were monitored on photo-diode array detector at a wavelength of 254 nm using a mixture (40:60) of buffer and acetonitrile. Solution concentrations were measured on a weight basis to avoid the use of an internal standard. The method was statistically validated for forced-degradation study, linearity and range, accuracy, precision, stability of analytical solutions, and selectivity. Due to its simplicity, rapid, and accuracy, we believe that the method will be useful to determine assay and known impurity of Celecoxib. PMID- 16221613 TI - Preparation and evaluation of SEDDS and SMEDDS containing carvedilol. AB - A new self-emulsifying drug delivery system (SEDDS) and self-microemulsifying drug delivery system (SMEDDS) have been developed to increase the solubility, dissolution rate, and, ultimately, oral bioavailability of a poorly water soluble drug, carvedilol. Ternary phase diagrams were used to evaluate the self emulsification and self-microemulsfication domains. The self-emulsification time following introduction into an aqueous medium under gentle agitation was evaluated. The minimum self-emulsification time was found at a Tween 80 content of 40%. The particle size distribution and zeta-potential were determined. Benzoic acid had a dual function, it improved the self-emulsification performance of SEDDS and SMEDDS in 0.1 N HCl and lead to a positively charged emulsion. The in vitro dissolution rate of carvedilol from SEDDS and SMEDDS was more than two fold faster compared with that from tablets. The developed SEDDS formulations significantly improved the oral bioavailability of carvedilol significantly, and the relative oral bioavailability of SEDDS compared with commercially available tablets was 413%. PMID- 16221614 TI - Determination of fexofenadine in tablets by capillary electrophoresis in free solution and in solution with cyclodextrins as analyte carriers. AB - Capillary electrophoresis (CE) methods for the determination of fexofenadine (FEX) in commercial pharmaceuticals were developed. It was demonstrated that FEX could be effectively analyzed in free solution cationic CE at low pH. Another analytical approach studied was based on cyclodextrin (CD) modified CE where highly charged CD derivatives served as analyte carriers. In this way, the separation range was spread to physiological pH region and a CE analysis of FEX, present actually in its zwitterionic form, could be accomplished. Several parameters affecting the separations were studied, including the type and concentration of carrier ion, counterion, analyte carrier, and pH of the buffer. The methods based on the free solution CE and CD-modified CE were compared each other, validated, and applied for the determination of FEX in tablets. PMID- 16221615 TI - Biodegradable microparticulates of beta-estradiol: preparation and in vitro characterization. AB - Beta-estradiol has been recommended for the long-term therapy of osteoporosis and its oral formulations are subjected to intensive first pass metabolism. The present investigation was aimed at preparing and characterizing biodegradable microparticles of beta-estradiol with polymers such as PLA, PLGA 85/15, PLGA 75/25, and their mixtures. The microparticles were prepared by solvent evaporation method using methylene chloride as a solvent and polyvinyl alcohol as a surfactant. The drug-polymer ratios were 1:3, 1:5, and 1:7. The prepared microparticles (twelve formulations) were tested for encapsulation efficiency and in vitro drug release in 50% methyl alcohol/phosphate buffer pH 7.4. The results showed that the encapsulation efficiency varied from 81 to 100% and the formulation fabricated from PLGA 85/15 (1:3) showed less burst and consistent long time release. This formulation when further characterized displayed irregular spherical shape with an average particle size of 72 microm. The crystallinity of the drug was reduced when investigated using X-ray diffractometry. No chemical interaction between the drug and the polymer was observed as evidenced by FT-IR analysis. The results indicated that beta estradiol biodegradable microparticles with PLGA 85/15 (1:3) could be a suitable approach for long term therapy of osteoporosis. PMID- 16221616 TI - Interconversion studies of betamethasone acetate polymorphs. AB - The polymorph interconversions of Betamethasone Acetate (BA) were studied under various pharmaceutical conditions, such as grinding, heating and suspending in water, based on differential scanning calorimetry, thermogravimetric analysis, and X-ray powder diffraction. There existed enantiotropic relationships between the three polymorphs of BA, which were named form II, Ialpha, and Ibeta work, respectively. It was concluded that form II was the most stable form when suspended in water. PMID- 16221617 TI - Oral sustained delivery of theophylline and cimetidine from in situ gelling pectin formulations in rabbits. AB - The aim of this study was to evaluate the potential of an in situ gelling pectin formulation as a vehicle for the oral sustained delivery of theophylline and cimetidine. In vitro studies demonstrated diffusion-controlled release of theophylline from 1, 1.5, and 2% w/v pectin gels. Release of this drug from 1.5% w/v pectin gels formed in situ in rabbit stomach was sustained over a period of 12 hours giving a theophylline bioavailability some seven fold higher than when administered from a commercial syrup. In contrast, interactions between cimetidine and pectin led to weak gelation of the pectin sols that prevented any meaningful determination of in vitro release characteristics. Similarly, in vivo release profiles from pectin formulations containing cimetidine were similar to that from a solution of this drug in buffer, indicative of weak gelation. Examination of the content of the rabbit stomach 5 hours after administration of 1.5% w/v pectin sols containing drug confirmed gel formation, but gels containing cimetidine were noticeably softer than those containing theophylline. PMID- 16221619 TI - Juvenile male sexual offenders: the quality of motivation system of assessment and treatment issues. AB - A group of juvenile male sexual offenders (n=100) completed the Quality of Motivation Questionnaire (QMQ) upon entry into a residential treatment facility. The concepts of Quality of Motivation (QM) Theory are presented to explain the QMQ scores. The scores include Disclosure Level, Sources of Motivation, Life Style Characteristics and Power. The results indicate abnormal motivation scores in the area of Disclosure Resistance, Depression, Primary and Learned Sources of Motivation, and all of the Maladaptive Skills Scores. Recommendations of treatment issues for therapists are prioritized according to QM Theory and presented in a treatment plan format called the Personal Development Plan (PDP). Implications for further research with the QMQ include outcome measurement of changes and comparison with non-offender groups. PMID- 16221618 TI - An evaluation of residential treatment for sexually aggressive youth. AB - This longitudinal study investigated the effectiveness of a one and one-half year long residential treatment program for 26 boys aged 11-15 with histories of sexually aggressive behavior. The clients were assessed with self-report and staff-report measures at the beginning and end of treatment and at several time points in between; also, aggressive acts were tabulated during a one-year follow up period. There was evidence of improved functioning on 10 of the 12 measures. During the follow-up, 27% of the sample committed an aggressive act, and 8% committed a sexually aggressive act. On most of the measures, improvement occurred gradually over the course of treatment, and on some measures most of the improvement occurred late in the placement. PMID- 16221620 TI - Conceptualizing child sexual abuse in interpersonal context: recovery of people, not memories. AB - The controversy regarding delayed recall of childhood sexual abuse (CSA) has deflected> attention from the question of whether memory recovery and other trauma-focused treatment approaches are effective for abuse survivors. More than a decade of clinical experience has convinced many experts that as a primary treatment strategy, this approach often leads to decompensation among CSA survivors. However, few treatment alternatives have been clearly articulated. One such alternative, contextual therapy, is grounded in evidence that many CSA survivors grow up in an interpersonal context that fails to transmit the capacities needed for effective daily functioning. Contextual therapy retains trauma-focused interventions as one component of a broader framework aimed at helping survivors develop adaptive capacities that, rather than having been disrupted by trauma, may never have been attained in the first place. From this perspective, abuse trauma is seen as compounding these deficits, rather than as a sole cause of them. PMID- 16221621 TI - Relationships of sexual, physical, and emotional abuse to emotional and behavioral problems among incarcerated adolescents. AB - This study examined the relationships of sexual, physical and emotional abuse to emotional (internalizing) and behavioral (externalizing) problems among incarcerated girls and boys. Participants were youth who were remanded to the correctional facilities within a statewide juvenile correctional system in a southern state in the United States of America. Each participant completed a structured interview regarding abuse history, emotional and behavioral difficulties, and demographic characteristics. Multiple regression analyses indicated that girls were more likely than boys to internalize their problems. The only abuse variable that was positively and significantly associated with emotional problems was emotional abuse. Greater behavioral problems were significantly related to youths being younger in age, white ethnicity, history of sexual abuse, and history of physical abuse. There were overall gender differences for internalizing problems, but not for externalizing problems among incarcerated adolescents. Furthermore, physical and sexual abuses were related to externalizing problems but not to internalizing problems. Thus, different types of abuse appear to have different effects on adolescent behavior. Implications for future research and practice are discussed. PMID- 16221622 TI - Patterns among sexual assault victims seeking treatment services. AB - The validity and reliability of research on the nature and extent of sexual assault tends to be affected by different definitions, methodologies, and measurements. As a result, two important aspects of sexual assault associated with patterns of symptom expression and therapeutic interventions are not often reflected in the research; the severity of the assault, including the duration of the abuse, and the age at the time of the assault and the gender of the victim. This research is based on intake forms from Hawai;i's only statewide provider of services to the victims of sexual assault. The analyses reveal that significant differences exist between male and female victims, by age and by assault characteristics, including the type of sexual assault, use of force and injury, length of assault, and the relationship between victim and offender. PMID- 16221623 TI - Partner abuse among women whose children have been sexually abused: an exploratory study. AB - To explore partner abuse among women in families in which a child has disclosed child sexual abuse, 102 mothers completed a measure of partner abuse and the Coping Responses Inventory. Results suggest that, like women in the general population, mothers of children who have been sexually abused have often experienced partner abuse. As expected, physical abuse in a current relationship was found to be related to the use of avoidance strategies in dealing with the sexual abuse disclosure. The implications for practitioners and for future research are discussed. PMID- 16221624 TI - Integrating cognitive-behavioral and expressive therapy interventions:applying the trauma outcome process in treating children with sexually abusive behavior problems. AB - This article presents intervention strategies based on the Trauma Outcome Process, an integrated treatment model for guiding clinical practice with children with sexually abusive behavior problems. The steps for completing a comprehensive assessment are reviewed, and strategies are presented to help clinicians create a therapeutic alliance and increase these children's self awareness of their thoughts, feelings, and body sensations. Treatment techniques based on an integrated approach combining cognitive-behavioral and expressive therapy approaches are described. These exercises help children accept responsibility for sexually inappropriate behavior and select adaptive responses for coping with the effects of traumatic experiences. PMID- 16221625 TI - What makes sex offenders confess? An exploratory study. AB - This study examines correlates of offender confession in criminal sexual conduct cases involving children. The cases consist of all closed court files (N=318), spanning the last 10 years from a single jurisdiction. This jurisdiction has a community-wide protocol for handling child sexual abuse cases, a high rate of charging (69%), a high rate of confession (64%), and high rates of pleas to sex crimes (77%). To determine what factors were associated with suspect confession before adjudication, we examined characteristics of the suspect, the child, the abuse, and the system using bi-variate and multi-variate analysis. The following four variables are associated with suspect confession: (1) having the state police conducting the law enforcement part of the investigation, (2) more serious abuse, (3) younger age of the suspect, and (4) having a court appointed (as opposed to a retained) attorney. PMID- 16221626 TI - Trauma specific versus generic measurement of distress and the validity of self reported symptoms in sexually abused children. AB - This study examined two issues in the assessment of child sexual abuse victims: sensitivity to trauma-related symptoms and validity of self-reports. The Trauma Symptom Checklist for Children (TSCC) and Personality Inventory for Youth (PIY) were completed by 41 sexually abused children. Parents and children completed diagnostic interviews for PTSD. We predicted that TSCC validity scales would be moderately correlated with PIY scales measuring similar constructs, and that TSCC clinical scales would be more sensitive to PTSD status than the PIY clinical scales. Results supported both hypotheses. TSCC validity scales appeared to be less likely to identify clinical cases, however. Clinical and research implications are discussed. PMID- 16221627 TI - A survey of problematic sexualized behaviors of children in the New York City Child Welfare System:estimates of problem, impact on services, and need for training. AB - Forty-eight of 52 (92%) agencies contracted with the New York City Administration for Children's Services (ACS) responded to a survey about problematic sexualized behaviors (PSB) of children in different levels of care within the child welfare system. Results revealed that almost all agencies reported PSB within their foster boarding home and residential treatment centers. A majority of agencies perceived PSB to be a significant problem for which staff and families were not sufficiently trained. These findings highlight many avenues for advocacy, clinical intervention, and staff development. PMID- 16221628 TI - Sexual abuse victimization and psychological distress among adolescent offenders. AB - This study focused on sexual abuse victimization and psychological distress among 272 adolescent offenders. The respondents were interviewed while they were being detained in a short-term holding facility. Female respondents reported more sexual abuse victimization and psychological distress than did their male counterparts. Furthermore, church attendance moderated the association between sexual abuse victimization and psychological distress among the male respondents. Implications of these findings for research and interventions with adolescent offenders are discussed. PMID- 16221629 TI - Exploring the multiplicity of childhood sexual abuse with a focus on polyincestuous contexts of abuse. AB - This study explored the prevalence of multiple abusers and polyincest in a clinical sample of 88 adult women, focusing on interviews with a subset of 30. For the sample as a whole, 43% reported three or more abusers and 23% reported polyincest. The narrative data was divided into three categories defined as Single-Abuser (N=8), Multiple Abuser (N=8), and Polyincestuous (N=14). These were compared to determine how survivors perceived family members' and others' reactions to disclosure of CSA. All three groups experienced various forms of silencing. The most negative responses were linked to polyincestuous abuse. Three themes were identified as denial/ minimization/normalization, fear/reprisal, and negative/inadequate. Results suggest the need for further research on the frequency of multiple abusers among victims of sexual abuse, as well as the ways in which family and social contexts serve as collusive environments for the practice of CSA. PMID- 16221630 TI - A study of the cycle of abuse among child molesters. AB - The current study was designed to determine whether there is support for the cycle of abuse theory among child molesters. A group of 147 sex offenders on probation were used in this study. The victims of 32 of these offenders were under the age of 10, and 19 of these offenders reported a history of childhood sexual abuse. Results indicate that being abused as a child is related to being a child molester. It was also found that the MMPI-2 did not predict the status of the subjects in regard to being abused as a child and having a victim under 10 years-old. PMID- 16221631 TI - Providing sexual education to victims of child sexual abuse:what is a clinician to do? AB - Despite the large body of research on childhood sexual abuse, virtually no one has examined the coverage of sexual education in treatment. Agencies from across the United States that specialize in treating child and adolescent victims of sexual abuse were surveyed. The results indicate that sexual education is covered in treatment with children of all ages, with male and female clients, and in both individual and group therapy. There was a statistically significant difference in the coverage of sexual education based on clients' age, but not based on gender or treatment modality. Parents are often included in treatment; however, the amount of parental involvement varies. Published materials, such as children's books and videos, are frequently used. Participants (i.e., clinicians) are satisfied with their coverage of sexual education in treatment even though clients often experience negative reactions. Results suggest that covering sexual education in treatment helps decrease some of the negative effects of sexual abuse. This study also serves as a test of Dillman's Total Design Method of conducting mail surveys. PMID- 16221632 TI - Authority as coercion:when authority figures abuse their positions to perpetrate child sexual abuse. AB - ABSTRACT. This article discusses child sexual abuse by a person in a position of authority, such as the child's teacher, guardian, relative, sports coach, or other person with authority over a child because of his/her particular position. The article tracks the recent trend toward recognizing position of authority in both state legislation and judicial precedent. Understanding the confusion and intimidation surrounding a child's experiences as a result of being sexually abused by a person in a position of authority often explains why children often fail to report or delay in reporting such abuse. Thus, existence of a perpetrator's position of authority in a particular case of child sexual abuse should influence a court's rulings on the elements of sexual abuse or assault in particular state statutes, as well as what evidence should be admissible. Ultimately, the author concludes that all states should recognize position of authority in their child abuse statutes, that such statutes should be interpreted broadly by the courts, and, finally, that evidence of the defendant's prior acts of sexual abuse should almost always be admissible at trial. PMID- 16221633 TI - Perceptions of child sexual abuse:victim and perpetrator characteristics, treatment efficacy, and lay vs. legal opinions of abuse. AB - Two studies examined interactions between victim age and victim response, and victim relation to perpetrator and victim response influencing perceptions of child sexual abuse (CSA). Undergraduates read one of several vignettes describing a sexual encounter between a man and a girl. In Experiment 1, age of the girl was varied; victim age interacted with victim response to increase ratings of abuse and expected trauma for the girl. In Experiment 2, age was held constant while victim relation to perpetrator was varied; men gave higher ratings of abuse than did women for scenarios involving a step father rather than a neighbor, regardless of victim response. In both studies, psychotherapy was expected to help the victim more than the perpetrator and the law was judged to be less stringent than it is regarding CSA. Results suggest that perceptions of CSA are influenced by several factors and that laws regarding CSA may not be well understood. PMID- 16221634 TI - Women's report of unwanted sexual attention during childhood. AB - Unwanted Sexual Attention (UWSA) refers to unsolicited verbal comments, gestures, stares, and other non-physical attention toward another's sexuality and physical appearance. The present study describes the occurrence of childhood UWSA and explores its relation to child sexual abuse (CSA). Three hundred fifteen university women (mean age=19.5) completed the Exposure to Sexual Attention Scale, a 200-item inventory which assesses various forms of UWSA received prior to age 18. Findings suggested that non-contact UWSA is a pervasive experience across settings for girls, with 98.7% reporting UWSA during childhood. Those reported most often to perpetrate UWSA (e.g., other students, adult strangers, child strangers, and other children) were also most often reported as sexual abusers. Emotional reactions by women to both UWSA and CSA were predominantly negative. Although significantly greater levels of fear reactions were reported by the CSA group when compared to the UWSA group. No difference was found between the two groups on levels of anger reactions to UWSA and CSA. Overall findings suggest that UWSA is an almost universal experience for girls that causes significant anger and fear. Assessing UWSA will be important from a public health perspective in evaluating if such behaviors pose a risk factor for negative developmental outcomes for girls. PMID- 16221635 TI - Child sexual abuse and attachment theory:are we rushing headlong into another controversy? AB - The child sexual abuse knowledge base has developed within a highly charged political and social context. As such, early theories of sexual abuse were susceptible to biases that reflected the larger sociocultural context. The purpose of this paper is to consider whether attachment theory as it is applied to child sexual abuse might also be susceptible to such biases. Although this paper concludes that attachment theory can potentially add an important dimension to the conceptualization of child sexual abuse and its dynamics, it is also suggested that attachment theory may remain at risk for becoming a vehicle for transmitting political and ideological agendas. PMID- 16221636 TI - Attachment theory and child abuse:some cautions. PMID- 16221637 TI - Young female sex offenders: assessment and treatment issues. AB - Research on the characteristics and treatment of juvenile female sex offenders is limited. This article reviews the literature on female offenders and presents the results of a nationwide survey by mail of 250 mental health providers in private settings and residential treatment centers (RTCs), on approaches to diagnosing prior sexual abuse history and/or perpetration among juvenile females. Nineteen of the most experienced practitioners were also interviewed by telephone to provide further insight to the issues highlighted by the surveys. Key findings include the lack of research, tools, and literature on young female sex offenders and perceived differences between male and female offenders including history, treatment, and characteristics. Further study and awareness of this population is encouraged to address clinical needs. PMID- 16221638 TI - The relationship of trauma exposure to sex offending behavior among male juvenile offenders. AB - The most common type of adult and juvenile sex offender treatment utilizes a Relapse Prevention (RP) model. In RP clients learn about their offense cycle with an emphasis on recognizing high-risk situations and negative emotional states that can be precursors or triggers to offending behavior. This study identifies ways that traumatic experiences and trauma-associated feelings can be offense triggers for juvenile sex offenders. Researchers interviewed the treating clinicians of 40 male juvenile sex offenders who received at least six months of RP sex offender treatment. Results showed that 95% of the youths had experienced a Post Traumatic Stress Disorder (PTSD) Criterion A traumatic event and that 65% met criteria for PTSD based on clinician judgments. Overall, clinicians identified prior trauma exposure as being related to the offense triggers in 85% of offenders. Specifically, the following trauma-related feelings were identified as offense triggers: intense fear in 37.5% of sex offenders, helplessness in 55%, and horror in 20%. Implications for sex offender treatment programs are discussed. PMID- 16221639 TI - Empathy, intimacy, loneliness and locus of control in child sex offenders: a comparison between familial and non-familial child sexual offenders. AB - There has been relatively little previous research examining differences between subgroups of child sex offenders. In this study, we compared a group of familial and non-familial child sex offenders attending a community based treatment program on a number of relationship dimensions thought to be important in explaining offending. No statistically significant differences were found between these groups on measures of general empathy, intimacy and loneliness, although non-familial offenders were found to have higher levels of cognitive empathy and a more internal locus of control than familial offenders. These findings are discussed in terms of their possible implications for service delivery. PMID- 16221640 TI - Reporting suspected child abuse: a pilot comparison of middle and high school counselors and principals. AB - A survey on experiences with child abuse reporting was obtained from 116 middle and high school counselors and principals across the U.S. Results revealed that counselors throughout their careers had made more reports than principals and perceived themselves to be better trained on child abuse. Those school professionals with fewer years of experience reported more adequate pre-service training. Counselors also failed to report in more instances than principals, citing as deterrents the lack of physical evidence and little faith in child protective services. Most counselors and principals took appropriate action when presented with case examples of child sexual abuse. Recommendations for future research are provided. PMID- 16221641 TI - The Colorado Sex Offender Risk Scale. AB - This study documents the development of an adult sex offender risk assessment tool. A sample of 494 sex offenders were followed for an average of 30 months. A risk scale was developed based upon criminal and therapeutic outcomes. The final risk scale included prior juvenile felony convictions, prior adult felony convictions, failure of the first or second grade, not being employed, victim being intoxicated, the perpetrator reporting not being sexually aroused during the crime, possession of a weapon during the crime, denial in therapy, sexual deviance in therapy, and motivation in therapy. The risk scale provided significant relative risk ratios against program failure at 12 and 30 months. Overall, those scoring high on the risk tool were 372% as likely to fail as those scoring low. PMID- 16221642 TI - Child sexual abuse in Sri Lanka: the current state of affairs and recommendations for the future. AB - This article focuses on child sexual abuse in Sri Lanka. The background of child sexual abuse in the country and its current status are discussed. The role of the National Child Protection Authority, the primary governmental body concerned with child protection, is also explored. The article also brings to light various cultural dimensions that serve to call attention to the issue of sexual abuse of children in Sri Lanka. These are explored and illustrated with case vignettes from the author's clinical experience. Finally, the article elucidates the main issues and barriers in Sri Lanka that need to be surmounted in order to have an integrated service delivery in the care of the sexually abused child. PMID- 16221644 TI - The trans-theoretical stages of change model and motivational interviewing for building maternal supportiveness in cases of sexual abuse. AB - When a child has been sexually abused, the non-offending parent plays a crucial role in determining that child's recovery. However, the disclosure of sexual abuse in families often throws non-offending parents into a dilemma about whether or not to side with the perpetrator or child, leaving them in an ambivalent position. In order to enhance the ability of parents to protect and support their children, this paper proposes the use of the Trans-theoretical Stages of Change model and Motivational Interviewing. Techniques of Motivational Interviewing will be discussed and then illustrated with a case example. PMID- 16221643 TI - Child (sexual) abuse: a universal problem, and Sri Lanka is no exception. PMID- 16221645 TI - Re-traumatization among adult women sexually abused in childhood: exploratory analyses in a prospective study. AB - The current exploratory study examined a variety of risk factors related to re traumatization, defined broadly, in a prospective sample of women who were sexually abused in childhood. Eighty women, who were part of a larger longitudinal study of the effects of child sexual abuse, were interviewed at three points in time: in childhood and at two points in early adulthood. Risk factors were measured at the second interview and used to predict reported trauma exposure between interviews two and three. Given theoretical arguments for differences in risk factors based on ecological context, analyses were conducted separately for married and unmarried women. Situational and intrapersonal risk factors such as homelessness and depression were predictive of re-traumatization risk while reported social support satisfaction was a protective factor. Reports of trauma exposure between interviews two and three were related to mental health symptoms. PMID- 16221646 TI - The association between suicidal ideation and childhood and adult victimization. AB - Despite the burgeoning literature on suicidal behavior, there are meager data available on the relationship between suicidal ideation and emotional abuse in childhood; adult physical or sexual victimization; and adult re-victimization in those with a history of childhood maltreatment. This study addressed these gaps in the literature. In a sample of 494 female undergraduates, current suicidal ideation was assessed dichotomously using an item from the SCL-90 and victimization was assessed retrospectively with validated multi-item measures. Analyses revealed an association between suicidal ideation and emotional and physical abuse in childhood and a history of partner violence. No relationship was found between childhood sexual abuse or forced sex in adulthood and suicidal ideation. Additionally, results of multiple logistic regression analyses did not support the hypothesis that the relationship between childhood victimization and SI is moderated by re-victimization in adulthood. PMID- 16221647 TI - Adolescent females who have sexually offended: comparisons with delinquent adolescent female offenders and adolescent males who sexually offend. AB - Very little is known about adolescent females who commit sexual offenses. In this two-part exploratory study, a sample of 11 adolescent females with sexual offense histories is described. In Study I, the sample is compared to an age-matched sample of 11 adolescent females with non-sexual victim-involved offense histories. The sexually offending group had significantly fewer antisocial behavior problems, such as alcohol or drug use problems, problems with fighting, or problems at school. They began their offense behaviors at younger ages than their non-sex offending delinquent peers. Few differences emerged with respect to attitudes about offense behaviors, such as level of denial. In Study II, the same sample of females with sex offense histories was compared with a group of age matched adolescent males with sex offense histories. The two sex-offending groups were found to be remarkably similar. There were few differences with respect to psychosocial and criminal histories, antisocial behavior, and variables related to clinical presentation and treatment (e.g., level of denial about offense). Likewise, the two groups are remarkably similar with respect to specific sex offense behaviors. The females, however, appear to have experienced more severe and pervasive abuse compared to the males. PMID- 16221648 TI - Further understanding the systemic effects of childhood sexual abuse: a comparison of two groups of clinical couples. AB - This study addressed the impact of traumatic experiences on dyadic relationships by comparing general stress and trauma symptoms and relationship impairment measures between two clinical groups: female childhood sexual abuse (CSA) survivors (n=15) and their male partners, and a control group of couples who reported no CSA (n=17). Both female CSA survivors and their partners reported higher symptoms of stress, suggesting support for the theory of secondary traumatic stress. Relationship impairment results did not support the hypothesis that CSA would negatively impact the dyadic functioning of couples. Clinical implications and recommendations for future research are provided. PMID- 16221649 TI - Relationships between maternal adult attachment security, child perceptions of maternal support, and maternal perceptions of child responses to sexual abuse. AB - This study assessed the relationships between maternal adult attachment style, children's perceptions of maternal support following disclosure of sexual abuse, and maternal perceptions of children's behavioral and emotional responses to sexual abuse among African-American child sexual abuse victims aged 4 to 12 (n=96) and a comparison group of non-abused subjects (n=100). Mothers with insecure attachment styles reported significantly higher rates of internalizing behaviors in their sexually abused children than did securely attached mothers. Among mothers of non-abused children, those with insecure adult attachment styles reported significantly higher rates of externalizing behaviors shown by their children in comparison with mothers with a secure adult attachment style. Mothers with insecure adult attachment styles also reported higher rates of overall behavior problems in their non-abused children that approached statistical significance. Sexually abused children's perceptions of maternal support were not related to maternal attachment style nor to child functioning. Contrary to our prediction, mothers of sexually abused children did not show lower rates of secure attachments when compared to mothers of non-sexually abused children. Our findings indicate that fostering parent- child attachment is important in order to decrease the risk for behavior problems and symptomatology in sexually abused children. PMID- 16221650 TI - Effects of group therapy on female adolescent survivors of sexual abuse: a pilot study. AB - Treatment interventions for female sexual abuse survivors was explored through a pilot study examining the relationship between group treatment and adolescent self-image. Self-image was defined as body image, self-reliance, self-control, and impulse-control. An experimental pre-post design was utilized. Participants were 13 female adolescent high school drop-outs with a history of sexual abuse who participated in the National Guard Youth Challenge Program at Camp Shelby in Mississippi. Participants completed the Offer Self- Image Questionnaire for Adolescents. The hypothesis that group therapy was an effective intervention for sexual abuse survivors was not supported; however, this is likely due to the small sample size. Because this was a pilot study, mean trends were observed to see directional changes that may assist future researchers. Observation of mean trends revealed that participants who received group therapy increased in levels of impulse control while the group that did not receive group therapy remained the same. Mean trends also revealed that the experimental group had a decrease in self-reliance whereas the control group maintained their levels of self-reliance. PMID- 16221651 TI - Child sexual abuse evaluations: adherence to recommendations. AB - Sixty-eight parents of children with alleged child sexual abuse (CSA) were surveyed 4 months after visiting an interdisciplinary CSA evaluation clinic in order to examine the extent to which recommendations were recalled and implemented. Of those parents for whom specific recommendations were provided, 9% recalled recommendations for medical follow-up, 79% recalled recommendations for child therapy, and 43% recalled recommendations for parental therapy. Families were more likely to receive and adhere to recommendations when there were behavioral problems and a strong indication of CSA. To improve communication with families, the reasons for recommendations must be clearly conveyed, and possible barriers to implementation should be anticipated and addressed. In addition, recommendations need to be realistic, and professionals need to facilitate the implementation of their treatment plan. PMID- 16221652 TI - Confronting abusers: the opinions of clinicians and survivors. AB - Confrontation of an abuser is often regarded as a powerful process in clinical work with adult survivors of childhood sexual abuse. This paper explores direct and symbolic forms of survivors confronting their abusers via a review of the current literature and the self-report of 12 female survivors, post-therapy. Six of the survivors had chosen to directly confront their abusers and six had not. Their experiences and opinions are presented together with a discussion of important issues within this area and the need for further research. PMID- 16221653 TI - Child sexual abuse victims' perception of paternal support as a significant predictor of coping style and global self-worth. AB - One hundred eighty-eight female participants (44 victims and 144 non-victims of CSA) from a 4-year university were recruited. The first objective examined the difference between victims and non-victims on measures of perceived social support, coping strategies, and global self-worth. Victims were more likely to report lower maternal support than non-victims of CSA. Additionally, victims who were perpetrated by a family member were more likely to report lower maternal support than those who were perpetrated by a non-family member. The second objective investigated the impact of victimization and several mediating factors on global self-worth. Victimization indirectly related to global self-worth through paternal support. Additional analyses examining non-victims showed significant direct relations between paternal, friend, and campus support leading to global self-worth. For victims, paternal support indirectly related to global self-worth through emotion- focused coping. These results indicate that paternal support and coping strategies may be important for later psychological adjustment in victims of CSA. PMID- 16221654 TI - Differences between sexually abused and non-sexually abused adolescent girls in foster care. AB - This descriptive study examines the differences between sexually abused and non sexually abused adolescent females in the foster care system who were participating in an independent living program. Fifty-four percent of the 190 girls met the criteria for being categorized as sexually abused. Those who experienced sexual abuse had also experienced significantly more of other types of child maltreatment. In addition, those who had been sexually abused were much more likely to be living in a congregate living setting, such as a group home or residential center, than those who were not sexually abused. The girls who had been sexually abused exhibited significantly more behavioral difficulties, including internalizing and externalizing problems, with 51% of them having clinically significant scores on the Youth Self-Report version of the Child Behavior Checklist. When co-occurrence of substance use and mental health problems were examined, sexually abused girls were significantly more likely than the non-sexually abused girls to meet the established criteria. PMID- 16221655 TI - Community corrections officers' attributions for sexual offending against children. AB - The present research examined gender differences in community corrections officers' (CCOs') attributions for child sexual offending. Eighty-five CCOs were asked to write down the reasons why they thought men sexually abused children, and then rate their reasons using Benson's Attributional Dimensions Scale. The results found that CCOs' reasons regarding why men sexually abuse children strongly paralleled current scientific theories on the etiology of child sexual abuse. Also, significant gender differences were found regarding the frequency with which participants cited certain types of reasons for child sexual abuse. Female CCOs were more likely to cite power and control as a reason, while male CCOs were more likely to cite psychopathology as a reason for child sexual abuse. No other gender differences were found. The research, clinical, and educational implications pertaining to these findings are discussed. PMID- 16221657 TI - Impaired sensory-emotional integration in a violent adolescent sex offender. AB - Social risk factors, executive neuropsychological functioning, and emotional numbing were examined as potential risk factors for violent sexual assaults by an adolescent male. The subject had been exposed to at least four previously identified social risk factors, including neglect, early separation from both parents, sexual abuse, and low socioeconomic status. At the time of assessment, executive neuropsychological functioning was intact. At the time of assessment, executive neuropsychological functioning was intact. While impaired emotional responsiveness was an important factor in the assaults, the construct of emotional numbing is too broad to identify the specific neuropsychological impairment involved in the subject's assault behavior. The authors suggest that failures in sensory-emotional integration contributed to the likelihood of violent behavior. Implications for further research and clinical practice are discussed. PMID- 16221658 TI - Multiple sexual victimizations among adolescent boys and girls: prevalence and associations with eating behaviors and psychological health. AB - The purpose of this study was to assess the prevalence of sexual abuse, including multiple victimizations, among adolescents and to examine associations among history of sexual abuse, disordered eating behaviors and psychological health. The sample included 81,247 students (40,946 girls and 40,301 boys) in 9th and 12th grade in Minnesota public schools. Sexual abuse was reported by 14.7% of girls and 6.2% of boys. Sexual abuse was associated with unhealthy eating behaviors, suicidal thoughts and attempts, and lower emotional well-being and self-esteem. Students who reported a single or multiple forms of sexual abuse were more likely than their non-abused peers to binge-eat (Odds Ratio: girls = 1.93-2.32; boys = 2.26-5.61), fast (OR: girls = 1.68-2.34; boys = 1.33-2.32), use diet pills (OR: girls = 1.50-4.30; boys = 2.99-17.29) or laxatives (OR: girls = 1.87-5.11; boys = 3.89-29.22), vomit (OR: girls = 1.75-4.06; boys = 2.82-24.16), and have suicidal thoughts/attempts (OR: girls = 3.01-6.12; boys = 3.35-9.46). Boys and girls reporting multiple sexual victimizations and had the highest odds ratios for disordered eating behaviors. Future research should explore strategies for primary prevention of revictimization and secondary prevention of detrimental effects of abuse. PMID- 16221659 TI - Predictors of maternal support: the point of view of adolescent victims of sexual abuse and their mothers. AB - Maternal support received by sexual abuse victims is considered a key factor in coping with the aftermath of abuse. The present study looked at four groups of potential predictors of maternal support: mothers' psychosocial characteristics, abuse characteristics, victim's characteristics, and disclosure characteristics. A total of 120 adolescents aged 12 to 17 years and their mothers, who were recruited from Child Protective Services, completed questionnaires and semi structured interviews. Multiple regression analyses performed separately on mother and child data sets revealed five significant predictors of maternal support in each analysis, four of which were common to both. The discussion underscores the importance of taking into account both the mother's and the victim's perceptions regarding psychological adjustment and family environment. PMID- 16221660 TI - Emotional closeness with perpetrators and amnesia for child sexual abuse. AB - Over the past decade, a contentious debate regarding delayed memories of childhood sexual abuse (CSA) has existed. In order to address this debate, 240 female participants completed questions about CSA, the Dissociative Experience Scale (Bernstein & Putnam, 1986), Perceived Emotional Closeness with Perpetrator Scale (Schultz, Passmore, & Yoder, 2000), and the Gudjonsson Suggestibility Scale 2 (GSS 2) (Gudjonsson, 1987). Eighty-two (36%) reported CSA and 37% of these indicated memory disturbances for the CSA. Participants reporting memory disturbances also reported significantly higher numbers of perpetrators, chemical abuse in their families, and closer relationships with the perpetrator(s) than participants reporting no memory disturbances. Implications for clinicians working with clients reporting CSA are discussed. PMID- 16221661 TI - Social reactions to child sexual abuse disclosures: a critical review. AB - Recent studies have examined disclosure of child sexual abuse to determine the correlates and consequences of telling others about this form of victimization. The present article reviews the current empirical literature on disclosure and reactions to adult survivors to assess what is known about the process of disclosure and whether telling others is therapeutic and leads to positive outcomes. Most studies assessing social reactions in detail have concerned adult survivors retrospectively reporting on their disclosures of child sexual abuse. Few empirical studies have been conducted in this area but research suggests that few victims tell anyone about child sexual abuse as children, and that the type of reactions to disclosure vary according to when disclosure occurs (childhood or adulthood), the extent and nature of the disclosure, and the person to whom one discloses. Clear evidence shows that negative social reactions are harmful to survivors' well-being, but better assessment of specific reactions and their effects are needed in theoretically-based studies to evaluate how these responses affect survivors' recovery in the context of other variables. Suggestions for future research on social reactions of others to adult survivors disclosing child sexual abuse are presented. PMID- 16221662 TI - Treatment of incarcerated, sexually-abused adolescent females: an outcome study. AB - This study examined the psychosocial functioning of 100 adolescent females (ages 12-17) sentenced to secure care in a southeastern state and the impact of gender specific, cognitive-behavioral therapy (CBT) intervention on the psychosocial functioning of subjects who reported a history of sexual abuse. The Multidimensional Adolescent Assessment Scale (MAAS) was used to assess psychosocial functioning. Pre-test scores on the MAAS revealed significantly higher scores on 12 of 16 dimensions of psychosocial functioning and higher rates of serious criminal behavior for youth who subsequently disclosed sexual abuse histories as compared to those without such histories. At post-test, statistically significant improvements in psychosocial functioning were observed on 14 of 16 MAAS subscales for those who received the CBT intervention. Thus, incarcerated female adolescents who reported a history of sexual abuse demonstrated more impairment in their functioning as compared to those without a reported history of sexual abuse and responded positively to gender-specific, CBT based intervention. PMID- 16221663 TI - Intracranial angioplasty & stenting for cerebral atherosclerosis: a position statement of the American Society of Interventional and Therapeutic Neuroradiology, Society of Interventional Radiology, and the American Society of Neuroradiology. PMID- 16221664 TI - Isolation of monomeric human V(H)s by a phage selection. AB - Human V(H) domains are promising molecules in applications involving antibodies, in particular, immunotherapy because of their human origin. However, they are, in general, prone to aggregation. Therefore, various strategies have been employed to acquire monomeric human V(H)s. We had previously discovered that filamentous phages displaying engineered monomeric V(H) domains gave rise to significantly larger plaques on bacterial lawns than phages displaying wild type V(H)s with aggregation tendencies. Using plaque size as the selection criterion and a phage displayed naive human V(H) library we identified 15 V(H)s that were monomeric. Additionally, the V(H)s demonstrated good expression yields, good refolding properties following thermal denaturation, resistance to aggregation during long incubation at 37 degrees C, and to trypsin at 37 degrees C. These 15 V(H)s should serve as good scaffolds for developing immunotherapeutics, and the selection method employed here should have general utility for isolating proteins with desirable biophysical properties. PMID- 16221665 TI - Lunatic fringe, manic fringe, and radical fringe recognize similar specificity determinants in O-fucosylated epidermal growth factor-like repeats. AB - Notch signaling is a component of a wide variety of developmental processes in many organisms. Notch activity can be modulated by O-fucosylation (mediated by protein O-fucosyltransferase-1) and Fringe, a beta1,3-N acetylglucosaminyltransferase that modifies O-fucose in the context of epidermal growth factor-like (EGF) repeats. Fringe was initially described in Drosophila, and three mammalian homologues have been identified, Manic fringe, Lunatic fringe, and Radical fringe. Here for the first time we have demonstrated that, similar to Manic and Lunatic, Radical fringe is also a fucose-specific beta1,3-N acetylglucosaminyltransferase. The fact that three Fringe homologues exist in mammals raises the question of whether and how these enzymes differ. Although Notch contains numerous EGF repeats that are predicted to be modified by O fucose, previous studies in our laboratory have demonstrated that not all O fucosylated EGF repeats of Notch are further modified by Fringe, suggesting that the Fringe enzymes can differentiate between them. In this work, we have sought to identify specificity determinants for the recognition of an individual O fucosylated EGF repeat by the Fringe enzymes. We have also sought to determine differences in the biochemical behavior of the Fringes with regard to their in vitro enzymatic activities. Using both in vivo and in vitro experiments, we have found two amino acids that appear to be important for the recognition of an O fucosylated EGF repeat by all three mammalian Fringes. These amino acids provide an initial step toward defining sequences that will allow us to predict which O fucosylated EGF repeats are modified by the Fringes. PMID- 16221666 TI - Apical transport and folding of prostate-specific membrane antigen occurs independent of glycan processing. AB - Prostate-specific membrane antigen (PSMA) is an integral cell-surface membrane glycoprotein that is overexpressed in prostate carcinomas rendering it an appropriate target for antibody-based therapeutic strategies. The biosynthesis of PSMA in transfected COS-1 cells reveals a slow conversion of mannose-rich to complex glycosylated PSMA compatible with slow transport kinetics from the endoplasmic reticulum to the Golgi. Importantly, mannose-rich PSMA persists as a trypsin-sensitive protein throughout its entire life cycle, and only Golgi located PSMA glycoforms acquire trypsin resistance. This resistance, used here as a tool to examine correct folding, does not depend on the type of glycosylation, because different PSMA glycoforms generated in the presence of inhibitors of carbohydrate processing in the Golgi are also trypsin resistant. The conformational transition of PSMA to a correctly folded molecule is likely to occur in the Golgi and does not implicate ER molecular chaperones, such as BiP. We show here that PSMA is not only heavily N-but also O-glycosylated. The question arising is whether glycans, which do not play a role in folding of PSMA, are implicated in its transport to the cell surface. Neither the cell-surface expression of PSMA nor its efficient apical sorting in polarized Madin-Darby canine kidney cells are influenced by modulators of N- and O-glycosylation. The acquisition of folding determinants in the Golgi, therefore, is an essential prerequisite for protein trafficking and sorting of PSMA and suggests that altered or aberrant glycosylation often occurring during tumorigenesis has no regulatory effect on the cell-surface expression of PSMA. PMID- 16221667 TI - Chromosomal instability induced by Pim-1 is passage-dependent and associated with dysregulation of cyclin B1. AB - Overexpression of the oncogenic serine/threonine kinase Pim-1 has been shown to induce chromosomal missegregation and polyploidy in prostate epithelial cell lines (1). Here we demonstrated that Pim-1-induced polyploidy develops in a passage-dependent manner in culture consistent with a stochastic mode of progression. Induction of chromosomal instability by Pim-1 was not restricted to prostate cells as it was also observed in telomerase-immortalized normal human mammary epithelial cells. Elevated levels of cyclin B1 protein, but not its messenger RNA, were evident in early passage Pim-1 overexpressing cells, suggesting that increased cyclin B1 levels contribute to the development of polyploidy. Furthermore, regulation of cyclin B1 protein and cyclin B1/CDK1 activity after treatment with anti-microtubule agents was impaired. Small interfering RNA targeting cyclin B1 reversed the cytokinesis delay but not the mitotic checkpoint defect in Pim-1 overexpressing cells. These results indicated that chronic Pim-1 overexpression dysregulates cyclin B1 protein expression, which contributes to the development of polyploidy by delaying cytokinesis. PMID- 16221668 TI - Crystal structure of the FERM domain of focal adhesion kinase. AB - Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that localizes to focal adhesions in adherent cells. Through phosphorylation of proteins assembled at the cytoplasmic tails of integrins, FAK promotes signaling events that modulate cellular growth, survival, and migration. The amino-terminal region of FAK contains a region of sequence homology with band 4.1 and ezrin/radixin/moesin (ERM) proteins termed a FERM domain. FERM domains are found in a variety of signaling and cytoskeletal proteins and are thought to mediate intermolecular interactions with partner proteins and phospholipids at the plasma membrane and intramolecular regulatory interactions. Here we report two crystal structures of an NH2-terminal fragment of avian FAK containing the FERM domain and a portion of the regulatory linker that connects the FERM and kinase domains. The tertiary folds of the three subdomains (F1, F2, and F3) are similar to those of known FERM structures despite low sequence conservation. Differences in the sequence and relative orientation of the F3 subdomain alters the nature of the interdomain interface, and the phosphoinositide binding site found in ERM family FERM domains is not present in FAK. A putative protein interaction site on the F3 lobe is masked by the proximal region of the linker. Additionally, in one structure the adjacent Src SH3 and SH2 binding sites in the linker associate with the surfaces of the F3 and F1 lobes, respectively. These structural features suggest the possibility that protein interactions of the FAK FERM domain can be regulated by binding of Src kinases to the linker segment. PMID- 16221669 TI - Novel antibody hinge regions for efficient production of CH2 domain-deleted antibodies. AB - HuCC49 deltaCH2 is a heavy chain constant domain 2 domain-deleted antibody under development as a radioimmunotherapeutic for treating carcinomas overexpressing the TAG-72 tumor antigen. Mammalian cell culture biosynthesis of HuCC49 deltaCH2 produces two isoforms (form A and form B) in an approximate 1:1 ratio, and consequently separation and purification of the desired form A isoform adversely impact process and yield. A protein engineering strategy was used to develop a panel of hinge-engineered HuCC49 deltaCH2 antibodies to identify hinge sequences to optimize production of the form A isoform. We found that adding a single proline residue at Kabat position 243, immediately adjacent to the carboxyl end of the core middle hinge CPPC domain, resulted in an increase from 39 to 51% form A isoform relative to the parent HuCC49 deltaCH2 antibody. Insertion of the amino acids proline-alanine-proline (PAP) at positions 243-245 enhanced production of the form A isoform to 72%. Insertion of a cysteine-rich 15-amino acid IgG3 hinge motif (CPEPKSCDTPPPCPR) in both of these mutant antibodies resulted in secretion of predominantly form A isoform with little or no detectable form B. Yields exceeding 98% of the form A isoform have been realized. Preliminary peptide mapping and mass spectrometry analysis suggest that at least two, and as many as five, inter-heavy chain disulfide linkages may be present. PMID- 16221670 TI - Conformational dimorphism of self-peptides and molecular mimicry in a disease associated HLA-B27 subtype. AB - An interesting property of certain peptides presented by major histocompatibility complex (MHC) molecules is their acquisition of a dual binding mode within the peptide binding groove. Using x-ray crystallography at 1.4 A resolution, we show here that the glucagon receptor-derived self-peptide pGR ((412)RRRWHRWRL(420)) is presented by the disease-associated human MHC class I subtype HLA-B*2705 in a dual conformation as well, with the middle of the peptide bent toward the floor of the peptide binding groove of the molecule in both binding modes. The conformations of pGR are compared here with those of another self-peptide (pVIPR, RRKWRRWHL) that is also displayed in two binding modes by HLA-B*2705 antigens and with that of the viral peptide pLMP2 (RRRWRRLTV). Conserved structural features suggest that the N-terminal halves of the peptides are crucial in allowing cytotoxic T lymphocyte (CTL) cross-reactivity. In addition, an analysis of T cell receptors (TCRs) from pGR- or pVIPR-directed, HLA-B27-restricted CTL clones demonstrates that TCR from distinct clones but with comparable reactivity may share CDR3alpha but not CDR3beta regions. Therefore, the cross-reactivity of these CTLs depends on TCR-CDR3alpha, is modulated by TCR-CDR3beta sequences, and is ultimately a consequence of the conformational dimorphism that characterizes binding of the self-peptides to HLA-B*2705. These results lend support to the concept that conformational dimorphisms of MHC class I-bound peptides might be connected with the occurrence of self-reactive CTL. PMID- 16221671 TI - Mammalian Smaug is a translational repressor that forms cytoplasmic foci similar to stress granules. AB - Cytoplasmic events depending on RNA-binding proteins contribute to the fine tuning of gene expression. Sterile alpha motif-containing RNA-binding proteins constitute a novel family of post-transcriptional regulators that recognize a specific RNA sequence motif known as Smaug recognition element (SRE). The Drosophila member of this family, dSmaug, triggers the translational repression and deadenylation of maternal mRNAs by independent mechanisms, and the yeast homologue Vts1 stimulates degradation of SRE-containing messengers. Two homologous genes are present in the mammalian genome. Here we showed that hSmaug 1, encoded in human chromosome 14, represses the translation of reporter transcripts carrying SRE motifs. When expressed in fibroblasts, hSmaug 1 forms cytoplasmic granules that contain polyadenylated mRNA and the RNA-binding proteins Staufen, TIAR, TIA-1, and HuR. Smaug 1 foci are distinct from degradation foci. The murine protein mSmaug 1 is expressed in the central nervous system and is abundant in post-synaptic densities, a subcellular region where translation is tightly regulated by synaptic stimulation. Biochemical analysis indicated that mSmaug 1 is present in synaptoneurosomal 20 S particles. These results suggest a role for mammalian Smaug 1 in RNA granule formation and translation regulation in neurons. PMID- 16221672 TI - ADAMTS13 substrate recognition of von Willebrand factor A2 domain. AB - ADAMTS13 controls the multimeric size of circulating von Willebrand factor (VWF) by cleaving the Tyr1605-Met1606 bond in theA2 domain. To examine substrate recognition, we expressed in bacteria and purified three A2 (VWF76-(1593-1668), VWF115-(1554-1668), VWFA2-(1473-1668)) and one A2-A3 (VWF115-A3-(1554-1874)) domain fragments. Using high pressure liquid chromatography analysis, the initial rates of VWF115 cleavage by ADAMTS13 at different substrate concentrations were determined, and from this the kinetic constants were derived (Km 1.61 microM; kcat 0.14 s(-1)), from which the specificity constant kcat/Km was calculated, 8.70 x 10(4) m(-1) s(-1). Similar values of the specificity constant were obtained for VWF76 and VWF115-A3. To identify residues important for recognition and proteolysis of VWF115, we introduced certain type 2A von Willebrand disease mutations by site-directed mutagenesis. Although most were cleaved normally, one (D1614G) was cleaved approximately 8-fold slower. Mutagenesis of additional charged residues predicted to be in close proximity to Asp1614 on the surface of the A2 domain (R1583A, D1587A, D1614A, E1615A, K1617A, E1638A, E1640A) revealed up to 13-fold reduction in kcat/Km for D1587A, D1614A, E1615A, and K1617A mutants. When introduced into the intact VWFA2 domain, proteolysis of the D1587A, D1614A, and E1615A mutants was also slowed, particularly in the presence of urea. Surface plasmon resonance demonstrated appreciable reduction in binding affinity between ADAMTS13 and VWF115 mutants (KD up to approximately 1.3 microM), compared with VWF115 (KD 20 nM). These results demonstrate an important role for Asp1614 and surrounding charged residues in the binding and cleavage of the VWFA2 domain by ADAMTS13. PMID- 16221673 TI - The structure of human SULT1A1 crystallized with estradiol. An insight into active site plasticity and substrate inhibition with multi-ring substrates. AB - Human SULT1A1 belongs to the supergene family of sulfotransferases (SULTs) involved in the sulfonation of xeno- and endobiotics. The enzyme is also one of the SULTs responsible for metabolic activation of mutagenic and carcinogenic compounds and therefore is implicated in various cancer forms. Further, it is not well understood how substrate inhibition takes place with rigid fused multiring substrates such as 17beta-estradiol (E2) at high substrate concentrations when subcellular fractions or recombinant enzymes are used. To investigate how estradiol binds to SULT1A1, we co-crystallized SULT1A1 with sulfated estradiol and the cofactor product, PAP (3'-phosphoadenosine 5'-phosphate). The crystal structure of SULT1A1 that we present here has PAP and one molecule of E2 bound in a nonproductive mode in the active site. The structure reveals how the SULT1A1 binding site undergoes conformational changes to accept fused ring substrates such as steroids. In agreement with previous reports, the enzyme shows partial substrate inhibition at high concentrations of E2. A model to explain these kinetics is developed based on the formation of an enzyme x PAP x E2 dead-end complex during catalysis. This model provides a very good quantitative description of the rate versus the [E2] curve. This dead-end complex is proposed to be that described by the observed structure, where E2 is bound in a nonproductive mode. PMID- 16221674 TI - FLN29, a novel interferon- and LPS-inducible gene acting as a negative regulator of toll-like receptor signaling. AB - Lipopolysaccharide (LPS) activates macrophages through toll-like receptor (TLR) 4. Although the mechanism of the TLR signaling pathway has been well documented, the mechanism of the negative regulation in response to LPS, particularly LPS tolerance, is still poorly understood. In this study we identified and characterized a novel interferon- and LPS-inducible gene, FLN29, which contains a TRAF6-related zinc finger motif and TRAF family member-associated NF-kappaB activator-related sequences. The induction of FLN29 was dependent on STAT1. The forced expression of FLN29 in macrophage-like RAW cells resulted in the suppression of TLR-mediated NF-kappaB and mitogen-activated protein kinase activation, while a reduced expression of FLN29 by small interfering RNA partly cancelled the down-regulation of LPS signaling. Furthermore, we demonstrated that NF-kappaB activation induced by TRAF6 and TAB2 was impaired by co-expression of FLN29, suggesting FLN29 may regulate the downstream of TRAF6. Taken together, FLN29 is a new negative feedback regulator of TLR signaling. PMID- 16221675 TI - Transgenic small interfering RNA halts amyotrophic lateral sclerosis in a mouse model. AB - Many autosomal dominant diseases such as familial amyotrophic lateral sclerosis (ALS) with copper/zinc superoxide dismutase (SOD1) mutation may be induced by missense point mutations that result in the production of proteins with toxic properties. Reduction in the encoding of proteins from such mutated genes can therefore be expected to improve the disease phenotype. The duplex of 21 nucleotide RNA, known as small interfering RNA (siRNA), has recently emerged as a powerful gene silencing tool. We made transgenic (Tg) mice with modified siRNA, which had multiple mismatch alternations within the sense strand, to prevent the "shutdown phenomenon" of transgenic siRNA. Consequently, the in vivo knockdown effect of siRNA on SOD1 expression did not diminish over four generations. When we crossed these anti-SOD1 siRNA Tg mice with SOD1G93A Tg mice, a model for ALS, siRNA prevented the development of disease by inhibiting mutant G93A SOD1 production in the central nervous system. Our findings clearly proved the principle that siRNA-mediated gene silencing can stop the development of familial ALS with SOD1 mutation. PMID- 16221676 TI - G betagamma binds histone deacetylase 5 (HDAC5) and inhibits its transcriptional co-repression activity. AB - In a yeast two-hybrid screen designed to identify novel effectors of the G betagamma subunit of heterotrimeric G proteins, we found that G betagamma binds to histone deacetylase 5 (HDAC5), an enzyme involved in a pathway not previously recognized to be directly impacted by G proteins. Formation of the G beta1gamma2 HDAC5 complex in mammalian cells can be blocked by overexpression of G alpha(o), and this inhibition is relieved by activation of alpha2A-adrenergic receptor, suggesting that the interaction occurs in a signal-dependent manner. The C terminal domain of HDAC5 binds directly to G betagamma through multiple motifs, and overexpression of this domain mimics the C terminus of G protein-coupled receptor kinase 2, a known G betagamma scavenger, in its ability to inhibit the G betagamma/HDAC5 interaction. The C terminus of HDAC4 shares significant similarity with that of HDAC5, and accordingly, HDAC4 is also able to form complexes with G beta1gamma2 in cultured cells, suggesting that the C-terminal domain of class II HDACs is a general G betagamma binding motif. Activation of a G(i/o)-coupled receptor results in a time-dependent activation of MEF2C, an HDAC5 regulated transcription factor, whereas inhibition of the interaction with a G betagamma scavenger inhibits MEF2C activity, suggesting a reduced potency of HDAC5-mediated inhibition. Taken together, these data imply that HDAC5 and possibly other class II HDACs can be added to the growing list of G betagamma effectors. PMID- 16221677 TI - The yeast Hsp110 Sse1 functionally interacts with the Hsp70 chaperones Ssa and Ssb. AB - There is growing evidence that members of the extended Hsp70 family of molecular chaperones, including the Hsp110 and Grp170 subgroups, collaborate in vivo to carry out essential cellular processes. However, relatively little is known regarding the interactions and cellular functions of Sse1, the yeast Hsp110 homolog. Through co-immunoprecipitation analysis, we found that Sse1 forms heterodimeric complexes with the abundant cytosolic Hsp70s Ssa and Ssb in vivo. Furthermore, these complexes can be efficiently reconstituted in vitro using purified proteins. Binding of Ssa or Ssb to Sse1 was mutually exclusive. The ATPase domain of Sse1 was found to be critical for interaction as inactivating point mutations severely reduced interaction with Ssa and Ssb. Sse1 stimulated Ssa1 ATPase activity synergistically with the co-chaperone Ydj1, and stimulation required complex formation. Ssa1 is required for post-translational translocation of the yeast mating pheromone alpha-factor into the endoplasmic reticulum. Like ssa mutants, we demonstrate that sse1delta cells accumulate prepro-alpha-factor, but not the co-translationally imported protein Kar2, indicating that interaction between Sse1 and Ssa is functionally significant in vivo. These data suggest that the Hsp110 chaperone operates in concert with Hsp70 in yeast and that this collaboration is required for cellular Hsp70 functions. PMID- 16221678 TI - Absence of a detectable intermediate in the compound I formation of horseradish peroxidase at ambient temperature. AB - A microsecond-resolved absorption spectrometer was developed to investigate the elementary steps in hydrogen peroxide (H(2)O(2)) activation reaction of horseradish peroxidase (HRP) at ambient temperature. The kinetic absorption spectra of HRP upon the mixing with various concentrations of H(2)O(2) (0.5-3 mm) were monitored in the time range from 50 to 300 mus. The time-resolved spectra in the Soret region possessed isosbestic points that were close to those between the resting state and compound I. The kinetic changes in the Soret absorbance could be well fitted by a single exponential function. Accordingly, no distinct spectrum of the putative intermediate between the resting state and compound I was identified. These results were consistent with the proposal that the O-O bond activation in heme peroxidases is promoted by the imidazolium form of the distal histidine that exists only transiently. It was estimated that the rate constant for the breakage of the O-O bond in H(2)O(2) by HRP is significantly faster than 1 x 10(4) s(-1). PMID- 16221679 TI - Double hexamer disruption and biochemical activities of Methanobacterium thermoautotrophicum MCM. AB - Methanobacterium thermoautotrophicum MCM (mtMCM) is a helicase required for DNA replication. Previous electron microscopy studies have shown mtMCM in several oligomeric forms. However, biochemical studies suggest that mtMCM is a dodecamer, likely a double hexamer (dHex). The crystal structure of the N-terminal fragment of mtMCM reveals a stable dHex architecture. To further confirm that the dHex is not an artifact of crystal packing of two hexamers, we investigated the relevance of the dHex by disrupting the hexamer-hexamer interactions seen in the crystal structure via site-directed mutagenesis and examining various biochemical activities of the mutants in vitro. Using a combination of biochemical and structural assays, we demonstrated that changing arginine to alanine at amino acid position 161 or the insertion of a six-aminoacid peptide at the hexamer hexamer interface disrupted dHex formation and produced stable single hexamers (sHex). Furthermore, we showed that the sHex mutants retained wild-type level of ATPase and DNA binding activities but had decreased helicase activity when compared with the wild type dHex protein. These biochemical properties of mtMCM are reminiscent of those of SV40 large T antigen, suggesting that the dHex form of mtMCM may be the active helicase for DNA unwinding during the bidirectional DNA replication. PMID- 16221680 TI - Polymorphism and double hexamer structure in the archaeal minichromosome maintenance (MCM) helicase from Methanobacterium thermoautotrophicum. AB - Methanobacterium thermoautotrophicum minichromosome maintenance complex (mtMCM), a cellular replicative helicase, is a useful model for the more complex eukaryotic MCMs. Biochemical and crystallographic evidence indicates that mtMCM assembles as a double hexamer (dHex), but previous electron microscopy studies reported only the presence of single heptamers or single hexamers (Pape, T., Meka, H., Chen, S., Vicentini, G., Van Heel, M., and Onesti, S. (2003) EMBO Rep. 4, 1079-1083; Yu, X., VanLoock, M. S., Poplawski, A., Kelman, Z., Xiang, T., Tye, B. K., and Egelman, E. H. (2002) EMBO Rep. 3, 792-797). Here we present the first three-dimensional electron microscopy reconstruction of the full-length mtMCM dHex in which two hexamers contact each other via the structurally well defined N terminal domains. The dHex has obvious side openings that resemble the side channels of LTag (large T antigen). 6-fold and 7-fold rings were observed in the same mtMCM preparation, but we determined that assembly as a double ring favors 6 fold structures. Additionally, open rings were also detected, which suggests a direct mtMCM loading mechanism onto DNA. PMID- 16221681 TI - Repair of formamidopyrimidines in DNA involves different glycosylases: role of the OGG1, NTH1, and NEIL1 enzymes. AB - The oxidatively induced DNA lesions 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyG) and 4,6-diamino-5-formamidopyrimidine (FapyA) are formed abundantly in DNA of cultured cells or tissues exposed to ionizing radiation or to other free radical-generating systems. In vitro studies indicate that these lesions are miscoding, can block the progression of DNA polymerases, and are substrates for base excision repair. However, no study has yet addressed how these lesions are metabolized in cellular extracts. The synthesis of oligonucleotides containing FapyG and FapyA at defined positions was recently reported. These constructs allowed us to investigate the repair of Fapy lesions in nuclear and mitochondrial extracts from wild type and knock-out mice lacking the two major DNA glycosylases for repair of oxidative DNA damage, OGG1 and NTH1. The background level of FapyG/FapyA in DNA from these mice was also determined. Endogenous FapyG levels in liver DNA from wild type mice were significantly higher than 8-hydroxyguanine levels. FapyG and FapyA were efficiently repaired in nuclear and mitochondrial extracts from wild type animals but not in the glycosylase-deficient mice. Our results indicated that OGG1 and NTH1 are the major DNA glycosylases for the removal of FapyG and FapyA, respectively. Tissue-specific analysis suggested that other DNA glycosylases may contribute to FapyA repair when NTH1 is poorly expressed. We identified NEIL1 in liver mitochondria, which could account for the residual incision activity in the absence of OGG1 and NTH1. FapyG and FapyA levels were significantly elevated in DNA from the knock-out mice, underscoring the biological role of OGG1 and NTH1 in the repair of these lesions. PMID- 16221682 TI - mTOR.RICTOR is the Ser473 kinase for Akt/protein kinase B in 3T3-L1 adipocytes. AB - The insulin-signaling pathway leading to the activation of Akt/protein kinase B has been well characterized except for a single step, the phosphorylation of Akt at Ser-473. Double-stranded DNA-dependent protein kinase (DNA-PK), ataxia telangiectasia mutated (ATM) gene product, integrin-linked kinase (ILK), protein kinase Calpha (PKCalpha), and mammalian target of rapamycin (mTOR), when complexed to rapamycin-insensitive companion of mTOR (RICTOR), have all been identified as playing a critical role in Akt Ser-473 phosphorylation. However, the apparently disparate results reported in these studies are difficult to evaluate, given that different stimuli and cell types were examined and that all of the candidate proteins have never been systematically studied in a single system. Additionally, none of these studies were performed in a classical insulin responsive cell type or tissue such as muscle or fat. We therefore examined each of these candidates in 3T3-L1 adipocytes. In vitro kinase assays, using different subcellular fractions of 3T3-L1 adipocytes, revealed that phosphatidylinositol 3,4,5-trisphosphate-stimulated Ser-473 phosphorylation correlated well with the amount of DNA-PK, mTOR, and RICTOR but did not correlate with levels of ATM, ILK, and PKCalpha. PKCalpha was completely absent from compartments with Ser-473 phosphorylation activity. Although purified DNA-PK could phosphorylate a peptide derived from Akt that contains amino acid Ser-473, it could not phosphorylate full-length Akt2. Vesicles immunoprecipitated from low density microsomes using antibodies directed against mTOR or RICTOR had phosphatidylinositol 3,4,5 trisphosphate-stimulated Ser-473 activity that was sensitive to wortmannin but not staurosporine. In contrast, immunopurified low density microsome vesicles containing ILK could not phosphorylate Akt on Ser-473 in vitro. Small interference RNA knockdown of RICTOR, but not DNA-PK, ATM, or ILK, suppressed insulin-activated Ser-473 phosphorylation and, to a lesser extent, Thr-308 phosphorylation in 3T3-L1 adipocytes. Based on our cell-free kinase and small interference RNA results, we conclude that mTOR complexed to RICTOR is the Ser 473 kinase in 3T3-L1 adipocytes. PMID- 16221684 TI - Ataxia-telangiectasia-mutated (ATM) is a T-antigen kinase that controls SV40 viral replication in vivo. AB - The structurally related ATM (ataxia-telangiectasia-mutated) and ATR (ATM-Rad3 related) protein kinases fulfill overlapping yet non-redundant functions as key regulators of cellular DNA damage responses. We recently showed that ATM phosphorylates the cyclic AMP response element-binding protein, CREB, following exposure to ionizing radiation (IR) and other DNA-damaging stimuli. Here, we show that a phospho-specific antibody recognizing the major ATM phosphorylation site in CREB cross-reacts with SV40 large tumor antigen (LTag), a multifunctional oncoprotein required for replication of the SV40 minichromosome. The relevant IR induced phosphorylation site in LTag recognized by phospho-CREB antibody was mapped to Ser-120. IR strongly induced the phosphorylation of Ser-120 in an ATM dependent manner in mouse embryo fibroblasts. Infection of African green monkey CV1 cells with SV40 resulted in the activation of ATM and phosphorylation of LTag and endogenous ATM substrates. Infection-induced LTag phosphorylation correlated with the onset of DNA replication, was ATM-dependent, and peaked when viral DNA levels reached their maximum. SV40 replication in CV1 cells required an intact LTag Ser-120 phosphorylation site and was inhibited following transfection with ATM small interfering RNA suggesting that ATM is required for optimal SV40 replication in primate cells. Our findings uncover a direct link between ATM and SV40 LTag that may have implications for understanding the replication cycle of oncogenic polyoma viruses. PMID- 16221683 TI - Evidence that HIV-1 reverse transcriptase employs the DNA 3' end-directed primary/secondary RNase H cleavage mechanism during synthesis and strand transfer. AB - We previously analyzed strand transfers catalyzed by human immunodeficiency virus, type 1 reverse transcriptase (RT) in a hairpin-containing RNA template system. In this system, RT produces a series of adjacent RNase H cuts before the hairpin base on the first, or donor template that clears a region of the donor, facilitating invasion by the second, or acceptor RNA. Here we analyze characteristics of the prominent cuts before the hairpin base and their role in strand transfers. Analysis of the template cleavage pattern during synthesis suggested that the RT performs DNA 3' end-directed primary and secondary cuts while paused at the hairpin base and that these cuts contribute to creation of the invasion site. RT catalyzed similar cleavages on a substrate representing a paused cDNA-template intermediate. DNA 3' end-directed secondary cuts, which require positioning of the polymerase active site downstream of the primer terminus, had previously not been specifically identified during synthesis. Our findings indicate that during synthesis DNA 3' end-directed primary and secondary cuts occur at pause sites. RT mutants with substitutions at the His(539) residue in the RNase H active site were defective in secondary cleavages. Analysis of the template cleavage pattern generated by the His(539) mutants during synthesis revealed inefficient cleavage at the invasion site, correlating with defects in strand transfer. Overall, results indicate RT can catalyze pause-associated DNA 3' end-directed primary and secondary cuts during synthesis and these cuts can contribute to strand transfer by creation of an invasion site. PMID- 16221686 TI - Genetic perturbation of glycolysis results in inhibition of de novo inositol biosynthesis. AB - In a genetic screen for Saccharomyces cerevisiae mutants hypersensitive to the inositol-depleting drugs lithium and valproate, a loss of function allele of TPI1 was identified. The TPI1 gene encodes triose phosphate isomerase, which catalyzes the interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde 3 phosphate. A single mutation (N65K) in tpi1 completely abolished Tpi1p enzyme activity and led to a 30-fold increase in the intracellular DHAP concentration. The tpi1 mutant was unable to grow in the absence of inositol and exhibited the "inositol-less death" phenotype. Similarly, the pgk1 mutant, which accumulates DHAP as a result of defective conversion of 3-phosphoglyceroyl phosphate to 3 phosphoglycerate, exhibited inositol auxotrophy. DHAP as well as glyceraldehyde 3 phosphate and oxaloacetate inhibited activity of both yeast and human myo inositol-3 phosphate synthase, the rate-limiting enzyme in de novo inositol biosynthesis. Implications for the pathology associated with TPI deficiency and responsiveness to inositol-depleting anti-bipolar drugs are discussed. This study is the first to establish a connection between perturbation of glycolysis and inhibition of de novo inositol biosynthesis. PMID- 16221685 TI - A prohormone convertase cleavage site within a predicted alpha-helix mediates sorting of the neuronal and endocrine polypeptide VGF into the regulated secretory pathway. AB - Distinct intracellular pathways are involved in regulated and constitutive protein secretion from neuronal and endocrine cells, yet the peptide signals and molecular mechanisms responsible for targeting and retention of soluble proteins in secretory granules are incompletely understood. By using confocal microscopy and subcellular fractionation, we examined trafficking of the neuronal and endocrine peptide precursor VGF that is stored in large dense core vesicles and undergoes regulated secretion. VGF cofractionated with secretory vesicle membranes but was not detected in detergent-resistant lipid rafts. Deletional analysis using epitope-tagged VGF suggested that the C-terminal 73-amino acid fragment of VGF, containing two predicted alpha-helical loops and four potential prohormone convertase (PC) cleavage sites, was necessary and sufficient with an N terminal signal peptide-containing domain, for large dense core vesicle sorting and regulated secretion from PC12 and INS-1 cells. Further transfection analysis identified the sorting sequence as a compact C-terminal alpha-helix and embedded 564RRR566 PC cleavage site; mutation of the 564RRR566 PC site in VGF-(1-65): GFP:VGF-(545-617) blocked regulated secretion, whereas disruption of the alpha helix had no effect. Mutation of the adjacent 567HFHH570 motif, a charged region that might enhance PC cleavage in acidic environments, also blocked regulated release. Finally, inhibition of PC cleavage in PC12 cells using the membrane permeable synthetic peptide chloromethyl ketone (decanoyl-RVKR-CMK) blocked regulated secretion of VGF. Our studies define a critical RRR-containing C terminal domain that targets VGF into the regulated pathway in neuronal PC12 and endocrine INS-1 cells, providing additional support for the proposed role that PCs and their cleavage sites play in regulated peptide secretion. PMID- 16221687 TI - Regulation of fatty acid synthesis by farnesyl pyrophosphate. AB - Fatty acid biosynthesis is transcriptionally regulated by liver X receptor (LXR) and its gene target, sterol regulatory element binding protein-1c (SREBP-1c). LXR activation is induced by oxysterol end products of the mevalonate pathway and is inhibited by the upstream non-sterol isoprenoid, geranylgeranyl pyrophosphate (GGPP). Whether isoprenoids play a role in regulating the transcription of genes involved in fatty acid biosynthesis is unknown. In CaCo-2 colon epithelial cells, depletion of mevalonate and its derivatives, including oxysterol ligands for LXR, increased fatty acid synthesis. Addition of mevalonate or its isoprenoid derivative, farnesyl pyrophosphate (FPP), prevented this increase. The effects of FPP were likely due to itself or its degradation products, because none of its downstream derivatives, GGPP, ubiquinone, or cholesterol, were effective. Moreover, the effects of FPP could not be accounted for by protein prenylation, because inhibition of farnesylation did not alter fatty acid synthesis in mevalonate-depleted cells incubated with the isoprenoid. Neither was fatty acid synthesis in these cells altered by inhibition of beta-oxidation. Mevalonate depletion increased fatty acid synthase (FAS) mRNA by transcriptional mechanisms, without increasing gene expression of other enzymes involved in fatty acid biosynthesis or of SREBP-1c. The abundance of mature SREBP-2 but not SREBP-1 was increased following mevalonate depletion. FPP prevented the increase in FAS mRNA in mevalonate-depleted cells without altering SREBP-2 activation. Thus, FPP regulates fatty acid synthesis by a mechanism that is likely independent of the SREBP pathway. PMID- 16221688 TI - Increased arterial stiffness in children on haemodialysis. AB - BACKGROUND: Measures of aortic stiffness--aortic pulse wave velocity (PWV) and augmentation index (AIx)--have been shown to be powerful predictors of survival in adult haemodialysis (HD) patients. Very few data have been reported regarding arterial stiffness in paediatric renal populations. METHODS: PWV and aortic AIx were determined from contour analysis of arterial waveforms recorded by applanation tonometry using a SphygmoCor device in 14 children on HD (age = 14.1 years) and in 15 age, height matched children controls. RESULTS: Pre-HD AIx (29.7 +/- 15.4%) and PWV (6.6 +/- 1.0 m/s) were significantly higher compared with children controls (8.3 +/- 8.0% and 5.4 +/- 0.6 m/s, respectively, P < 0.0001). The only significant difference between normal and HD children was BP level: 103/61 vs 114/72 mmHg, P < 0.05. In children of HD patients, a multiple linear regression model including BP, age, height, weight, Ca and P levels as independent variables accounted for 57% of the variability in AIx. Dialysis had no impact on AIx (post-HD: 28.5 +/- 12.7%) or on PWV (post-HD: 6.7 +/- 0.8 m/s). CONCLUSIONS: We show, in this first-ever report of increased arterial stiffness in children on dialysis, that end-stage renal disease is associated with abnormalities in arterial wall elastic properties, comparable with adult levels, even in childhood. Most importantly, the absence of a discernible amelioration with dialysis implies that purely structural and not functional alterations lie behind the increased arterial stiffness. PMID- 16221689 TI - Long-term use of vitamin E-coated polysulfone membrane reduces oxidative stress markers in haemodialysis patients. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase and an independent predictor of overall mortality and cardiovascular outcome in haemodialysis (HD) patients. In the present study, we compared the effects of a vitamin E-coated polysulfone membrane (PSE) and a non vitamin E-coated polysulfone membrane (PS) on oxidative stress markers such as ADMA. METHODS: Thirty-one HD patients were enrolled to this investigation. They were allocated into two groups: in the PSE group (n = 16), PSE was used for 6 months, followed by PS for an additional 12 months; in the PS group (n = 15), PS was used for the entire observation period. Plasma ADMA, oxidized low density lipoprotein (Ox-LDL) and malondialdehyde LDL (MDA-LDL) levels were measured at baseline, 3, 6, 12 and 18 months. Plasma ADMA in peritoneal dialysis (PD) patients and in healthy individuals was also measured. RESULTS: Predialysis concentrations of ADMA (0.72+/- 0.13 nmol/ml) were significantly higher in the HD group than in both PD patients (0.63+/-0.10 nmol/ml, P<0.01) and healthy individuals (0.44+/-0.01 nmol/ml, P<0.0001). Treatment with PSE for 6 months significantly reduced predialysis levels of ADMA (0.54+/-0.09 nmol/ml) compared with baseline (0.74+/-0.12 nmol/ml; P<0.01). Predialysis levels of Ox-LDL and MDA LDL after 6 months therapy with PSE were also significantly lower than baseline values. Treatment with PS subsequent to treatment with PSE again increased ADMA, Ox-LDL and MDA-LDL back to baseline levels. In the PS group, ADMA, Ox-LDL and MDA LDL levels remained unchanged during the entire treatment period of 18 months. CONCLUSIONS: We confirmed that use of PSE reduced ADMA that had accumulated in HD patients. This finding indicates that PSE exerts anti-oxidant activity. A randomized controlled study will be required to determine whether PSE prevents cardiovascular diseases and other dialysis-related complications by reducing oxidative stress. PMID- 16221690 TI - Minimal change nephrotic syndrome associated with systemic lupus erythematosus. PMID- 16221691 TI - Erythropoietin in the critically ill: what is the evidence? PMID- 16221692 TI - Effects of CAPD on hepatosteatosis and lipid profile. PMID- 16221693 TI - Insulin analogue usage in a haemodialysis patient with type 2 diabetes mellitus. PMID- 16221694 TI - Kimura's disease in a patient with idiopathic dilated cardiomyopathy on ambulatory peritoneal dialysis. PMID- 16221695 TI - Cinacalcet and vascular calcifications induced by calcitriol. PMID- 16221696 TI - The association between BP and mortality in patients on chronic peritoneal dialysis. PMID- 16221697 TI - Beneficial effects of icodextrin on plasma level of adipocytokines in peritoneal dialysis patients. AB - OBJECTIVE: Leptin and adiponectin, well-recognized adipocytokines, are reported to contribute to the pathogenesis of atherosclerosis. The aim of this study was to elucidate the effects of icodextrin-based dialysis solution on adipocytokine metabolism. METHODS: In 12 non-diabetic anuric patients on peritoneal dialysis, dialysis solution was changed from glucose-based dialysis solution to icodextrin based dialysis solution for 6 months. Plasma levels of leptin, adiponectin, lipids (total cholesterol, HDL-cholesterol and triglyceride), insulin, blood glucose and insulin sensitivity index by the homeostasis model assessment (HOMA IR) were compared before and after the use of the icodextrin solution. RESULTS: Plasma leptin level was decreased from 15.6 (2.5-69.0) to 7.3 (2.9-45.9) ng/ml (P = 0.018) and plasma adiponectin level increased from 11.6 (6.2-19.6) to 17.6 (7.8 33.0) microg/ml (P = 0.002). A reduction in plasma insulin level from 33.1 (13.8 54.1) to 19.1 (5.8-37.3) muU/ml (P = 0.009) and HOMA-IR from 8.22 (3.68-15.09) to 5.15 (1.40-13.78) (P = 0.015) was observed. While plasma total cholesterol level remained similar, HDL-cholesterol level increased, from 36.0 (22-45) to 43.5 (30 69) mg/dl (P = 0.008) and the triglyceride level decreased, from 174.0 (140-250) to 116.5 (81-207) mg/dl (P = 0.012). CONCLUSION: Icodextrin-based dialysis solution improves abnormal adipocytokine metabolism, dyslipidaemia and insulin resistance, which are known to be associated with atherosclerosis. These results suggest that the use of icodextrin-based dialysis solution might be useful in preventing atherosclerosis in PD patients. Long-term effects of icodextrin-based dialysis solution on the atherosclerosis in peritoneal dialysis patients should be tested. PMID- 16221698 TI - Influenza vaccine-induced rhabdomyolysis leading to acute renal transplant dysfunction. PMID- 16221699 TI - Tubular expression of connective tissue growth factor correlates with interstitial fibrosis in type 2 diabetic nephropathy. PMID- 16221700 TI - Alpha-galactosidase activity should be examined in patients with proteinuria: what have we learned from a family affected with Fabry disease? PMID- 16221701 TI - The role of non-haemodynamic factors of the genesis of LVH. PMID- 16221702 TI - Diagnostic potential of circulating natriuretic peptides in chronic kidney disease. AB - BACKGROUND: Measurement of natriuretic peptides, particularly brain natriuretic peptide (BNP) is an established method for the diagnosis of cardiovascular disorders, chiefly left ventricular (LV) dysfunction. The influence of renal function on the diagnostic utility of natriuretic peptides is unclear. METHODS: We performed a cross-sectional study of 296 patients with renal disease but no history of cardiac disease using echocardiography to assess LV mass and function. Circulating levels of atrial natriuretic peptide (ANP) and BNP were also measured. RESULTS: The incidence of LV hypertrophy increased with progressive renal dysfunction; from 39% in patients with near-normal renal function, to 80% in renal transplant patients. There was a negative correlation between both ANP and BNP, and glomerular filtration rate (GFR) (ANP: r = -0.28, P<0.001; BNP: r = 0.40, P<0.001). Serum ANP and BNP had sensitivity and specificity for LV hypertrophy of 39.9%, 87.4% (ANP) and 61.4%, 67.6% (BNP) respectively. Sensitivity and specificity for LV dysfunction was 77.2%, 32.4% (ANP) and 71.8%, 40.0% (BNP). Significant confounders in determining serum ANP were haemoglobin, beta blockade and albumin, while serum BNP levels were significantly confounded by GFR, albumin, haemoglobin, beta blockade and age. CONCLUSIONS: Across a spectrum of renal dysfunction, GFR is a more important determinant of serum BNP than ventricular function, and several factors are predictors of natriuretic peptide levels. In chronic kidney disease, the use of natriuretic peptides to diagnose LV hypertrophy must be interpreted in light of these other factors. The use of these peptides in renal dysfunction to diagnose LV dysfunction may be of limited value. PMID- 16221703 TI - The vitamin D prodrugs 1alpha(OH)D2, 1alpha(OH)D3 and BCI-210 suppress PTH secretion by bovine parathyroid cells. AB - BACKGROUND: Active vitamin D compounds are widely used in the treatment of secondary hyperparathyroidism associated with renal failure. These compounds reduce PTH secretion through vitamin D receptor (VDR)-dependent repression of PTH gene transcription. In previous studies, 1alpha(OH)D3, a vitamin D prodrug, inhibited PTH secretion in cultured bovine parathyroid cells, but it was unclear whether 1alpha(OH)D3 itself or an active metabolite produced this inhibition. METHODS: We determined the effectiveness of the vitamin D prodrugs 1alpha(OH)D3, 1alpha(OH)D2 and 1alpha(OH)-24(R)-methyl-25-ene-D2 (BCI-210) at inhibiting PTH secretion in bovine parathyroid cell cultures, and examined the metabolism of [3H]1alpha(OH)D2 in these cells. RESULTS: All three prodrugs suppressed PTH secretion with approximately 10% of the activity of 1,25(OH)2D3; much higher activity than expected based on the VDR affinities of these prodrugs (0.25% of 1,25(OH)2D3). Parathyroid cells activated [3H]1alpha(OH)D2 to both 1,25(OH)2D2 and 1,24(OH)2D2. 1,24(OH)2D2 was detectable at 4 h, increased to a maximum at 8 h, and then decreased. In contrast, 1,25(OH)2D2 levels increased linearly with time, suggesting the presence of constitutively active vitamin D-25-hydroxylase not previously reported in parathyroid cells. The cytochrome P-450 inhibitor ketoconazole (50 microM) reduced 1alpha(OH)D2 metabolism to below detectable levels, but did not significantly affect suppression of PTH by 1alpha(OH)D2. CONCLUSIONS: The vitamin D prodrugs 1alpha(OH)D3, 1alpha(OH)D2 and BCI-210 suppressed PTH production by cultured parathyroid cells. The ability of 1alpha(OH)D2 to reduce PTH despite inhibition of its metabolism suggests a direct action of this 'prodrug' on the parathyroid gland, but the mechanism underlying this activity is not yet known. PMID- 16221704 TI - The importance of standardization of creatinine in the implementation of guidelines and recommendations for CKD: implications for CKD management programmes. AB - BACKGROUND: In an attempt to reduce late referral and to improve the care of patients with chronic kidney disease (CKD), different organizations have issued guidelines on when to refer patients to the nephrologist. Most suggest referral of patients with a GFR below 60 ml/min/1.73 m2, and demand referral if the GFR is below 30 ml/min/1.73 m2. It is recommended to use the abbreviated MDRD equation to estimate GFR. This formula is, however, sensitive to the creatinine assay methodology. In addition, the impact of the implementation of such guidelines on the nephrology practice has never been evaluated. This study (i) identifies the true burden of CKD in a population and simulates the effects of a 100% implementation of the guidelines on the nephrology work load, and (ii) evaluates the validity of the estimated GFR using the abbreviated MDRD formula when routinely provided. METHODS: Different laboratories (both hospital and private) in our region were asked to report on all the serum creatinine values performed during the first week of December 2004. If patients had more than one determination, only the lowest serum creatinine value was retained. Patients already known to a nephrology unit were not included. GFR was calculated using the abbreviated MDRD, using the serum creatinine as reported by these laboratories, or after correction to the MDRD-standard using different published equations. RESULTS: 20,108 patients, with a mean age of 53.4+/-16.2 years, 48% females, had at least one serum creatinine determination in the observation period. According to the K/DOQI CKD classification, 20.2, 1.6 and 0.8% of females and 13.3, 1.6 and 0.6% of males were in stage 3, 4 and 5, respectively, when the abbreviated MDRD formula was used with the serum creatinine value as reported by the laboratories. Important differences in classifications were obtained when the different correction formulae for creatinine were applied. According to the current recommendations, this would lead to a mandatory referral of 1650-2400 CKD stage 4 patients per 100 000 inhabitants and a suggested referral of another 4100 15 360 CKD stage 3 patients per 100,000 inhabitants to a nephrology unit. CONCLUSION: Implementation of the current guidelines for referral of CKD patients to nephrologists would lead to an overload of the nephrology care capacities. Large differences in estimated GFRs with different corrections for serum creatinine are observed, resulting in important CKD classification differences. Standardization of serum creatinine assays is mandatory before guidelines, and especially the routine provision of the estimated GFR by the abbreviated MDRD formula, can be implemented in clinical practice. PMID- 16221705 TI - Pyroglutamic acidosis in a renal transplant patient. PMID- 16221706 TI - Metastatic intracranial subdural empyema from renal cyst infection in autosomal dominant polycystic kidney disease. PMID- 16221707 TI - Kaposi's sarcoma occurring in a young black man after kidney transplantation. PMID- 16221708 TI - Inhibition of mTOR with sirolimus slows disease progression in Han:SPRD rats with autosomal dominant polycystic kidney disease (ADPKD). AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by dysregulated tubular epithelial cell growth, resulting in the formation of multiple renal cysts and progressive renal failure. To date, there is no effective treatment for ADPKD. The mammalian target of rapamycin (mTOR) is an atypical protein kinase and a central controller of cell growth and proliferation. We examined the effect of the mTOR inhibitor sirolimus (rapamycin) on renal functional loss and cyst progression in the Han:SPRD rat model of ADPKD. METHODS: Five-week-old male heterozygous cystic (Cy/+) and wild-type normal (+/+) rats were administered sirolimus (2 mg/kg/day) orally through the drinking water for 3 months. The renal function was monitored throughout the treatment phase, and rats were sacrificed thereafter. Kidneys were analysed histomorphometrically, and for the expression and phosphorylation of S6K, a well-characterized target of mTOR in the regulation of cell growth. RESULTS: The steady increase in BUN and creatinine in Cy/+ rats was reduced by 39 and 34%, respectively with sirolimus after 3 months treatment. Kidney weight and 2-kidney/total body weight (2K/TBW) ratios were reduced by 34 and 26% in sirolimus-treated Cy/+ rats. Cyst volume density was also reduced by 18%. Of importance, Cy/+ rats displayed enhanced levels of total and phosphorylated S6K. Sirolimus effectively reduced total and phosphorylated levels of S6K. CONCLUSION: We conclude that oral sirolimus markedly delays the loss of renal function and retards cyst development in Han:SPRD rats with ADPKD. Our data also suggest that activation of the S6K signalling pathway plays an important role in the pathogenesis of PKD. Sirolimus could be a useful drug to retard progressive renal failure in patients with ADPKD. PMID- 16221709 TI - Erythropoietin attenuates renal injury in experimental acute renal failure ischaemic/reperfusion model. AB - BACKGROUND: Erythropoietin (EPO), originally identified for its critical role in promoting erythrocyte survival and differentiation, has been shown to exert multiple paracrine/autocrine functions. Protective effects of EPO have been demonstrated in various tissues and experimental models of ischaemia-induced injury. In the present study, we investigated the effect of EPO on an in vivo rat model of renal ischaemia/reperfusion (I/R) injury and the possible mechanisms implicated in the EPO-mediated anti-apoptotic action. METHODS: Male Wistar rats, subjected to renal ischaemia for 45 min, were administered either saline or EPO (500 U/kg, i.p.) 20 min prior to I/R. A sham-operated group served as the control. At 48 h of reperfusion, the renal dysfunction and injury was assessed by measurement of serum biochemical markers (urea, creatinine) and histological grading. Apoptosis was assessed by the TUNEL method and morphological criteria. Expression of Bax and NF-kappaB (p65) was also evaluated. RESULTS: High levels of serum urea and creatinine were identified at 48 h after ischaemia. The EPO treated group had significantly lower serum and creatinine levels. Semi quantitative assessment of the histological lesions showed that rats subjected to I/R developed marked structural damage, whereas significantly less tubular damage was observed in the EPO-treated group. I/R caused an increase in TUNEL-positive cells that was accompanied by morphological evidence of apoptosis. In the EPO treated rats only a few scattered TUNEL-positive cells were observed. Up regulation of Bax in the tubular epithelial cells and increased expression of NF kappaB was observed in the I/R-treated rats, while diminished expression of Bax and positive immunostaining of NF-kappaB was observed in the EPO-treated rats. CONCLUSION: Administration of EPO as a single dose before the onset of ischaemia produced a significant reduction in tubular injury, which was accompanied by a marked amelioration of renal functional impairment. The cytoprotective action of EPO against I/R injury seems to be associated with its anti-apoptotic action. Moreover, transcription factor NF-kappaB is likely to play a pivotal role in the pathophysiology of I/R renal injury and might have a key role in EPO-mediated protective effects. PMID- 16221710 TI - Effect of haemodialysis on signal-averaged electrocardiogram P-wave parameters. AB - BACKGROUND: The P-wave signal-averaged electrocardiogram (SAECG) is a non invasive technique considered to indicate an increased risk for paroxysmal atrial fibrillation. The study was designed to evaluate the effect of the haemodialysis (HD) process on SAECG parameters in the group of selected HD patients. METHODS: Forty-seven HD patients (without relevant cardiac diseases) were included. SAECGs were performed pre- and post-dialysis together with evaluating extracellular body water (ECW) by using bioimpedance and biochemical measurements. For each SAECG, filtered P-wave duration (FPD) and root mean square voltage of the final 20 ms of filtered P-wave (RMS20) were established. RESULTS: The duration of either pre- or post-dialysis FDP was higher in HD patients than in the control group (P<0.001 and P = 0.005, respectively). The voltage of either pre- or post-dialysis RMS20 was reduced in HD patients compared with controls (P<0.001 in both cases). HD induced a decrease in the duration of the FDP and a significant increase in the voltage of RMS20 (P<0.001 in both cases). Stepwise multiple regression identified independent predictors of pre- and post-dialysis FDP as: (1) age; (2) pre- and post-dialysis ECW/kg body weight, respectively and; (3) pre- and post-dialysis haemoglobin levels, respectively. In the case of RMS20, we did not find any independent predictors either pre- or post-dialysis. CONCLUSIONS: Our study revealed that P-wave SAECG parameters are abnormal in a significant portion of HD patients and improved with HD process. We have also demonstrated that patients' age, volume status as well as the presence of anaemia are important factors influencing P-wave SAECG parameters in HD patients. PMID- 16221711 TI - Immuno-allergic interstitial nephritis related to fluindione: first biopsy proven cases. PMID- 16221712 TI - Chronic allograft nephropathy: expression and localization of PAI-1 and PPAR gamma. AB - BACKGROUND: Chronic allograft nephropathy (CAN) is a major cause of loss of renal allografts. Mechanisms postulated to be involved include sequelae of rejection, warm ischaemia time, drug toxicity, ongoing hypertension and dyslipidaemia. Plasminogen activator inhibitor-1 (PAI-1) is implicated not only in thrombosis, but also in fibrosis, by inhibiting matrix degradation, and is expressed in renal parenchymal cells as well as in macrophages. Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a member of the steroid receptor superfamily, and plays a major beneficial role in lipid regulation, insulin sensitivity and macrophage function, factors that may play a role in CAN. We therefore studied the expression of these molecules in CAN. METHODS: All renal biopsy/nephrectomy files from Vanderbilt and Nashville VAMC from a 6 year period were reviewed to identify all renal transplant biopsies or nephrectomies more than 6 months after transplant with CAN. CAN was defined as fibrosis in the graft, vascular, interstitial or glomerular. All cases were scored for severity of fibrosis in vasculature (0-3 scale), glomeruli (% affected with either segmental and/or global sclerosis) and interstitial fibrosis (% of sample affected). PAI-1 and PPAR-gamma immunostaining was assessed on a 0-3 scale in glomeruli, vessels and tubules. RESULTS: Eighty-two patients with a total of 106 samples met entry criteria. The population consisted of 59 Caucasians and 23 African-Americans; 49 males, 33 females with average age 37.9+/-1.7 years. Average time after transplant at time of biopsy was 60.5+/-4.9 months (range 7-229). Glomerulosclerosis extent in CAN was on average 26.5+/-2.4% compared with 3.6+/ 1.2% in normal control kidneys from native kidney cancer nephrectomies and 0% in transplanted kidney biopsies from patients obtained > or =6 months after transplantation without CAN. Native control kidneys showed mild interstitial fibrosis (8.0+/-1.2%), whereas transplant controls showed very minimal fibrosis (2.0+/-2.0%). Interstitial fibrosis in CAN kidneys was on average 47.9+/-2.4%. Glomerular PAI-1 and PPAR-gamma staining scores were markedly increased in CAN (1.8+/-0.1, 2.3+/-0.1, respectively) compared with normal control kidneys from native kidney cancer nephrectomies (PAI-1 0.2+/-0.2 and PPAR-gamma 0.4+/-0.2, P<0.001) and transplanted kidney biopsies from patients obtained > or =6 months after transplantation without CAN (PAI-1 0 and PPAR-gamma 0, P<0.001). Tubular PAI-1 and PPAR-gamma staining scores were 1.9+/-0.1 and 1.9+/-0.1, respectively, and also increased over both native and transplant kidney controls (0.8+/-0.2 for both categories for PAI-1, 1.2+/-0.2 for both categories for PPAR-gamma, respectively). Vascular sclerosis in CAN was 1.0+/-0.1 with increased PAI-1 and PPAR-gamma scores (1.7+/-0.1, 1.2+/-0.1, respectively) compared with controls. Infiltrating macrophages were increased in CAN, and were positive for both PAI-1 and PPAR-gamma. Biopsies with less sclerosis overall showed a trend for less PAI 1 and PPAR-gamma staining. CONCLUSION: PAI-1 and PPAR-gamma are both increased in CAN compared with non-scarred native or transplant control kidneys. We speculate that altered matrix metabolism and macrophage function might be involved in the development of CAN. PMID- 16221713 TI - The leukotriene B4 receptor antagonist ONO-4057 inhibits mesangioproliferative changes in anti-Thy-1 nephritis. AB - OBJECTIVE: ONO-4057 is a specific leukotriene B4 (LTB4) receptor antagonist which inhibits human neutrophil aggregation, chemotaxis and degranulation induced by LTB4. This study was conducted to evaluate the role of LTB4 in glomerulonephritis, and to examine whether ONO-4057 moderated anti-Thy-1 nephritis. METHODS: Experiment 1: Sixty Wistar rats were divided into three groups. Rats of Group A (n = 20) underwent intraperitoneal administration of placebo as a control group, rats of Group B (n = 20) first underwent intraperitoneal administration of 100 mg/kg ONO-4057 and rats of Group C (n = 20) first underwent intraperitoneal administration of 300 mg/kg ONO-4057 daily from day 3 before anti-Thy-1 antibody (OX7) injection to day 14 after OX7 injection, respectively. Experiment 2: Forty rats were divided into two groups. ONO-group (n = 20) was treated with 300 mg/kg BW of ONO-4057 and placebo-group (n = 20) with placebo daily from days 1 to 13 after OX7 injection. Urine and blood samples were collected and the kidneys were extirpated from five rats of each group sacrificed at 3 h, 24 h, day 7 or day 14 after the injection of OX7 in both experiments. Urinary protein excretion, renal function and pathological findings were analysed in each group of both experiments. RESULTS: (1) Glomerular infiltration by polymorphonuclear leucocytes (PMNs) and macrophages at 3 h was less in Groups B and C than in Group A, and matrix scores at day 7 were lower in Groups B and C than in Group A. Injury scores did not differ among the groups. (2) Urinary protein excretion at day 7 was less in Group C than in Group A. (3) Neither pathological findings nor urinary protein excretion differed between ONO-group and placebo-group. CONCLUSION: These results suggest that LTB4 is associated not with the pathogenesis of complement-dependent mesangial cell lysis but with that of mesangial proliferative change in anti-Thy-1 nephritis. PMID- 16221714 TI - Acute respiratory infection in a renal transplant recipient. PMID- 16221715 TI - The unrecognized prevalence of chronic kidney disease in diabetes. AB - BACKGROUND: Diabetes mellitus and chronic kidney disease (CKD) are common and exhibit synergistic associations with premature mortality. Current diabetes guidelines in the UK recommend annual urinary albumin and serum creatinine determinations to screen for diabetic kidney disease. The aim of this study was to estimate the burden of CKD in patients with diabetes and examine the ability of serum creatinine and albuminuria to detect clinically meaningful CKD compared with estimated glomerular filtration rate (eGFR). METHODS: All adults known to have diabetes in primary and secondary care in Salford, UK, alive with independent renal function on 1 January 2004 were included in this observational study (n=7596). Demographic and laboratory parameters were obtained from the Electronic Patient Record. eGFR was determined using the 4-variable modification of diet in renal disease (MDRD) formula. Clinically meaningful CKD was defined as an eGFR <60 ml/min/1.73 m(2). RESULTS: Creatinine and albuminuria were measured in the preceding 2 years in 82.3 and 55.2% of subjects, respectively. In patients with CKD, normoalbuminuria was present in 48.8%, and serum creatinine was normal (or=120 micromol/l) had a sensitivity and specificity of 45.3 and 100%, respectively, to identify CKD. The combination of abnormal creatinine and albuminuria had an improved performance but still failed to detect a large number with CKD (sensitivity 82.4%, specificity 75.4%). Serum creatinine failed to identify CKD more often in females (OR 8.22, CI 6.56-10.29). CONCLUSIONS: Undiagnosed CKD is common in diabetes. Current screening strategies, based on creatinine or albuminuria, fail to identify a considerable number of subjects with CKD. Incorporating eGFR into screening for CKD would identify individuals earlier in the natural history of the disease and enable early effective treatment to delay progression of CKD. PMID- 16221716 TI - Transplantation of kidney with retrocaval ureter: what are the pitfalls? PMID- 16221717 TI - Incidence of chronic kidney disease in India. PMID- 16221718 TI - Gitelman syndrome: genetic and expression analysis of the thiazide-sensitive sodium-chloride transporter in blood cells. AB - Gitelman syndrome is caused by mutations of the SLC12A3 gene, which encodes the thiazide-sensitive NaCl transporter NCCT. Although several mutations causing Gitelman syndrome have been described, their molecular consequences have been rarely studied. We report a patient with Gitelman syndrome due to a mutation in the GT donor splicing site of intron 9. The analysis of RNA from peripheral blood cells showed a complete deletion of exon 9. This case report confirms the feasibility of using readily accessible blood cells to study the expression of the SLC12A3 gene, a procedure that may facilitate further studies of the functional genomics of Gitelman syndrome. PMID- 16221719 TI - Left ventricular function in patients with chronic kidney disease evaluated by colour tissue Doppler velocity imaging. AB - BACKGROUND: Cardiovascular disease is the leading cause of death in chronic kidney disease (CKD) patients. Tissue Doppler velocity imaging (TVI) is a new objective method that accurately quantifies myocardial tissue velocities, deformation, time intervals and left ventricular (LV) filling pressure. In this study, TVI was compared with conventional echocardiography for the assessment of left ventricular (LV) function in pre-dialysis patients with different stages of CKD. The results obtained by TVI were used to analyse possible relationships between LV function and clinical factors such as hyperparathyroidism and hypertension that could influence LV function. METHODS: Conventional echocardiography and TVI images were recorded in 40 patients (36 men and 4 women, mean age 60+/-14 years, range 28-80 years) and in 27 healthy controls (21 men and 6 women, mean age 58+/-17 years, range 28-82 years). Twenty-two patients had mild/moderate CKD (CCr>29 ml/min; Group 1) and 18 patients had severe CKD (CCr